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Sample records for e1 two-phonon strengths

  1. Reply to "Comment on two-phonon gamma-vibrational strength in osmium nuclei"

    SciTech Connect

    Wu, C.Y.; Cline, D.; Hayes, A.B.; Simon, M.W.; Krueken, R.; Cooper, J.R.; Barton, C.J.; Beausang, C.W.; Bialik, C.; Caprio, M.A.; Casten, R.F.; Hecht, A.A.; Newman, H.; Novak, J.; Pietralla, N.; Zyromski, K.; Zamfir, N.V.

    2002-09-03

    The claim that the two-phonon gamma-vibrational configuration constitutes a major component for the I=4+ states in osmium nuclei is based on solid experimental evidence. A non-negligible two-quasiparticle or hexadecapole component must also exist in order to explain the data.

  2. Quenching of the E1 strength in 149Nd

    NASA Astrophysics Data System (ADS)

    Ruchowska, E.; Płóciennik, W. A.; Mach, H.; Gulda, K.; Fogelberg, B.; Gausemel, H.; Fraile, L. M.; Kurcewicz, W.; Mezilev, K.; Sanchez-Vega, M.

    2010-07-01

    Lifetime measurements of excited states in 149Nd have been performed using the advanced time-delayed β γ γ( t) method. Half-lives of 14 excited states in 149Nd have been determined for the first time or measured with higher precision. Twelve new γ -lines and 5 new levels have been introduced into the decay scheme of 149Pr based on results of the γ γ coincidence measurements. Reduced transition probabilities have been determined for 40 γ -transitions in 149Nd . Configuration assignments for 6 rotational bands in 149Nd are proposed. Enhanced E1 transitions indicate that the ground-state band and the band built on the 332.9keV level constitute a pair of the ensuremath K^{π} = 5/2^{±} parity doublet bands. Potential energy surfaces on the ensuremath (β2,β3) -plane have been calculated for the lowest single quasi-particle configurations in 149Nd using the Strutinski method and the axially deformed Woods-Saxon potential. The predicted occurrence of the octupole-deformed K = 5/2 configuration is in agreement with experiment. Unexpectedly low ensuremath \\vert D_0\\vert values obtained for the ensuremath K^{π} = 5/2^{±} parity doublet bands may result from cancellation between the proton and neutron shell correction contributions to ensuremath \\vert D_0\\vert.

  3. Spectral collapse via two-phonon interactions in trapped ions

    NASA Astrophysics Data System (ADS)

    Felicetti, S.; Pedernales, J. S.; Egusquiza, I. L.; Romero, G.; Lamata, L.; Braak, D.; Solano, E.

    2015-09-01

    Two-photon processes have so far been considered only as resulting from frequency-matched second-order expansions of light-matter interaction, with consequently small coupling strengths. However, a variety of novel physical phenomena arises when such coupling values become comparable to the system characteristic frequencies. Here, we propose a realistic implementation of two-photon quantum Rabi and Dicke models in trapped-ion technologies. In this case, effective two-phonon processes can be explored in all relevant parameter regimes. In particular, we show that an ion chain under bichromatic laser drivings exhibits a rich dynamics and highly counterintuitive spectral features, such as interaction-induced spectral collapse.

  4. E 1 and M 1 γ -strength functions in 144Nd

    NASA Astrophysics Data System (ADS)

    Voinov, A. V.; Grimes, S. M.

    2015-12-01

    Both E 1 and M 1 γ -strength functions below the neutron separation energy were analyzed based on experimental data from 143Nd(n ,γ )144Nd and 143Nd(n ,γ α )140Ce reactions. It is confirmed that the commonly adopted E 1 model based on the temperature dependence of the width of the giant dipole resonance works well. The popular M 1 strength function due to the spin-flip magnetic resonance located near the neutron binding energy is not capable of reproducing experimental data. The low-energy enhancement of the M 1 strength or the energy-independent model of Weisskopf, both leading to the low-energy strength sizable to E 1 one, fit experimental data best.

  5. B(E1) strengths and isospin symmetry in {sup 67}As and {sup 67}Se

    SciTech Connect

    Orlandi, R.; De Angelis, G.; Della Vedova, F.; Gadea, A.; Napoli, D. R.; Recchia, F.; Valiente-Dobon, J. J.; Marginean, N.; Wiedemann, K. T.; Sahin, E.; Bracco, A.; Leoni, S.; Tonev, D.; Wadsworth, R.; Singh, B. S. Nara; Sarantites, D. G.; Reviol, W.; Chiara, C. J.; Pechenaya, O.; Lister, C. J.

    2007-11-30

    In a recent experiment at Argonne National Laboratory, new spectroscopic information was obtained for the mirror pair {sup 67}As and {sup 67}Se via the measurement of the lifetime of analogue 9/2{sub I}{sup +} excited states. The nuclei of interest were populated via the fusion evaporation reaction of {sup 32}S on {sup 40}Ca, and selected by the combination of Gammasphere, Microball and the Neutron Wall arrays. The mirror B(E1) strengths were measured via the centroid shift method in analogue E1 transitions in {sup 67}As and {sup 67}Se. These findings are significant in determining the extent of conservation of isospin symmetry.

  6. B(E1) Strengths from Coulomb excitation of 11Be

    SciTech Connect

    Summers, N C; Pain, S D; Orr, N A; Catford, W N; Angelique, J C; Ashwood, N I; Bouchat, V; Clarke, N M; Curtis, N; Freer, M; Fulton, B R; Hanappe, F; Labiche, M; Loucey, J L; Lemmon, R C; Mahboub, D; Ninane, A; Normand, G; Nunes, F M; Soic, N; Stuttge, L; Timis, C N; Thompson, I; Winfield, J S; Ziman, V

    2007-03-06

    The B(E1;1/2{sup +}{yields} 1/2{sup -}) strength for {sup 11}Be has been extracted from intermediate energy Coulomb excitation measurements, over a range of beam energies using a new reaction model, the extended continuum discretized coupled channels (XCDCC) method. In addition, a measurement of the excitation cross section for {sup 11}Be+{sup 208}Pb at 38.6 MeV/nucleon is reported. The B(E1) strength of 0.105(12) e{sup 2}fm{sup 2} derived from this measurement is consistent with those made previously at 60 and 64 MeV/nucleon, in contrast to an anomalously low result obtained at 43 MeV/nucleon. By coupling a multi-configuration description of the projectile structure with realistic reaction theory, the XCDCC model provides for the first time a fully quantum mechanical description of Coulomb excitation. The XCDCC calculations reveal that the excitation process involves significant contributions from nuclear, continuum, and higher-order effects. An analysis of the present and two earlier intermediate energy measurements yields a combined B(E1) strength of 0.105(7) e{sup 2}fm{sup 2}. This value is in good agreement with the value deduced independently from the lifetime of the 1/2{sup -} state in {sup 11}Be, and has a comparable precision.

  7. Giant two-phonon Raman scattering from nanoscale NbC precipitates in Nb

    NASA Astrophysics Data System (ADS)

    Cao, C.; Tao, R.; Ford, D. C.; Klie, R. F.; Proslier, T.; Cooley, L. D.; Dzyuba, A.; Zapol, P.; Warren, M.; Lind, H.; Zasadzinski, J. F.

    2015-03-01

    High-purity niobium (Nb), subjected to the processing methods used in the fabrication of superconducting rf cavities, displays micrometer-sized surface patches containing excess carbon. High-resolution transmission electron microscopy and electron energy-loss spectroscopy measurements are presented which reveal the presence of nanoscale NbC coherent precipitates in such regions. Raman backscatter spectroscopy on similar surface regions exhibit spectra consistent with the literature results on bulk NbC but with significantly enhanced two-phonon scattering. The unprecedented strength and sharpness of the two-phonon signal has prompted a theoretical analysis, using density functional theory (DFT), of phonon modes in NbC for two different interface models of the coherent precipitate. One model leads to overall compressive strain and a comparison to ab initio calculations of phonon dispersion curves under uniform compression of the NbC shows that the measured two-phonon peaks are linked directly to phonon anomalies arising from strong electron-phonon interaction. Another model of the extended interface between Nb and NbC, studied by DFT, gives insight into the frequency shifts of the acoustic and optical mode density of states measured by first-order Raman spectroscopy. The exact origin of the stronger two-phonon response is not known at present but it suggests the possibility of enhanced electron-phonon coupling in transition-metal carbides under strain found either in the bulk NbC inclusions or at their interfaces with Nb metal. Preliminary tunneling studies using a point contact method show some energy gaps larger than expected for bulk NbC.

  8. Two-phonon octupole excitation in /sup 147/Gd

    SciTech Connect

    Kleinheinz, P.; Styczen, J.; Piiparinen, M.; Blomqvist, J.; Kortelahti, M.

    1982-05-24

    A half-life measurement gives (57 +- 13)B/sub W/ for the 1575-keV E3 transition in /sup 147/Gd, which characterizes the 2.572-MeV (19/2)/sup -/ level as ..nu..f/sub 7/2/ x 3/sup -/ x 3/sup -/ two-phonon octupole state. This is the first identification of a nuclear two-phonon octupole excitation. The observed departures from harmonic vibration (twice the one-phonon energy and twice the transition rate) can be quantitatively understood in terms of the microscopic composition of the states, and can be derived from other observed features of the octupole vibrations in this region.

  9. Search for two-phonon vibrations in /sup 168/Er

    SciTech Connect

    Kleppinger, E.W.; Yates, S.W.

    1983-08-01

    The low-lying level structure of /sup 168/Er has been examined by the (n,n'..gamma..) reaction and all known levels with J<7 and E/sub x/< or =2.0 MeV are observed. The discovery of a level at 1893 keV calls for a reexamination of the completeness of the level scheme assumed previously for this nucleus. No new low-lying candidates for two-phonon ..gamma.. vibrations are observed.

  10. New probe of M1 and E1 strengths in GDR regions

    NASA Astrophysics Data System (ADS)

    Hayakawa, T.; Ogata, K.; Miyamoto, S.; Mochizuki, T.; Horikawa, K.; Amano, S.; Imazaki, K.; Li, D.; Izawa, Y.; Chiba, S.

    2014-05-01

    The M1 strengths (or level density of 1+ states) are of importance for estimation of interaction strengths between neutrinos and nuclei for the study of the supernova neutrino-process. In 1957, Agodi predicted theoretically angular distribution of neutrons emitted from states excited via dipole transitions with linearly polarized gamma-ray beam at the polar angle of θ=90° should be followed by a simple function, a + b cos(2φ), where φ, is azimuthal angel. However, this theoretical prediction has not been verified over the wide mass region except for light nuclei as deuteron. We have measured neutron angular distributions with (polarized gamma, n) reactions on Au, Nal, and Cu. We have verified the Agodi's prediction for the first time over the wide mass region. This suggests that (polarized gamma, n) reactions may be useful tools to study M1 strengths in giant resonance regions.

  11. New probe of M1 and E1 strengths in GDR regions

    SciTech Connect

    Hayakawa, T.; Ogata, K.; Miyamoto, S.; Mochizuki, T.; Horikawa, K.; Amano, S.; Imazaki, K.; Li, D.; Izawa, Y.; Chiba, S.

    2014-05-02

    The M1 strengths (or level density of 1{sup +} states) are of importance for estimation of interaction strengths between neutrinos and nuclei for the study of the supernova neutrino-process. In 1957, Agodi predicted theoretically angular distribution of neutrons emitted from states excited via dipole transitions with linearly polarized gamma-ray beam at the polar angle of θ=90° should be followed by a simple function, a + b cos(2φ), where φ, is azimuthal angel. However, this theoretical prediction has not been verified over the wide mass region except for light nuclei as deuteron. We have measured neutron angular distributions with (polarized gamma, n) reactions on Au, Nal, and Cu. We have verified the Agodi's prediction for the first time over the wide mass region. This suggests that (polarized gamma, n) reactions may be useful tools to study M1 strengths in giant resonance regions.

  12. Systematic calculation of E1 strength with fully self-consistent Skyrme-HF plus RPA

    SciTech Connect

    Inakura, Tsunenori; Yabana, Kazuhiro; Nakatsukasa, Takashi

    2010-08-12

    Systematic calculation of the electric dipole strength in even-even nuclei are performed employing a Hartree-Fock plus RPA approach. For an easy implementation of the fully self-consistent calculation, a finite amplitude method which we proposed recently is employed. We show an importance of low-angular momentum orbit for emergence of low-lying dipole resonances which have a substantial influence on the r-process in supernovae.

  13. ON THE QUESTIONS OF THE NUCLEAR LEVEL DENSITY AND THE E1 PHOTON STRENGTH FUNCTIONS

    SciTech Connect

    MUGHABGHAB,S.F.; DUNFORD,C.L.

    1999-11-15

    New results were derived from average level spacings of neutron resonances for the spin dispersion parameter of the nuclear level density, which demonstrated the influence of shell effects, as well as the interplay of nucleon pairing correlations for nuclei in the mass range from {sup 29}Si to {sup 241}Pu. The volume and surface components of the nuclear level density parameter, as well as the shell-damping factor, were determined as, a{sub v} = 0.076 {+-} 0.009 MeV{sup {minus}1} , a{sub s} = 0.180 {+-} 0.047 MeV{sup {minus}1}, and y{sub 0} = 0.047 {+-} 0.04 MeV{+-}, respectively. The effective nucleon mass at the Fermi surface is derived as m*/m = 1.09 {+-} 0.13. New evidence is presented for a dipole-quadrupole interaction term in the primary E1 transitions of average resonance capture data. This evidence is obtained by testing a proposed generalized Landau Fermi liquid model for spherical and deformed nuclei, which includes the effect of the dipole-quadrupole interaction. The Landau-Migdal interaction constant and the effective nucleon mass, are determined as F{sub 0}{prime} = 1.49 {+-} 0.08, and m*/m=1.04 {+-} 0.07, respectively.

  14. Two-Phonon Quantum Coherences in Indium Antimonide Studied by Nonlinear Two-Dimensional Terahertz Spectroscopy.

    PubMed

    Somma, Carmine; Folpini, Giulia; Reimann, Klaus; Woerner, Michael; Elsaesser, Thomas

    2016-04-29

    We report the first observation of two-phonon quantum coherences in a semiconductor. Two-dimensional terahertz (THz) spectra recorded with a sequence of three THz pulses display strong two-phonon signals, clearly distinguished from signals due to interband two-photon absorption and electron tunneling. The two-phonon coherences originate from impulsive off-resonant excitation in the nonperturbative regime of light-matter interaction. A theoretical analysis provides the relevant Liouville pathways, showing that nonlinear interactions using the large interband dipole moment generate stronger two-phonon excitations than linear interactions. PMID:27176538

  15. Fragmentation of the two-phonon octupole vibrational states in {sup 208}Pb

    SciTech Connect

    Vetter, K.; Macchiavelli, A.O.; Asztalos, S.J.; Busse, B.C.; Clark, R.M.; Deleplanque, M.A.; Diamond, R.M.; Fallon, P.; Kruecken, R.; Lee, I.Y.; MacLeod, R.W.; Schmid, G.J.; Stephens, F.S.; Cline, D.; Gray, R.; Simon, M.W.; Wu, C.Y.; Amro, H.; Janssens, R.V.; Amro, H.; Moore, E.F.

    1998-11-01

    An experiment designed to populate two-phonon vibrational states in {sup 208}Pb by Coulomb excitation was performed with a {sup 136}Xe beam at a bombarding energy of 650 MeV. The {gamma} rays from the decay of the excited states were measured with Gammasphere and scattered particles were detected in the compact heavy-ion counter CHICO. We have not been able to observe any state close to the expected harmonic energy of 5.2 MeV. However, we were able to extract the B(E3,3{sub 1}{sup {minus}}{r_arrow}6{sub 1}{sup +}) value for the lowest known 6{sup +} state at 4.424 MeV based on measured {gamma}-ray intensities. About 20{percent} of the expected total E3 strength can be found in this state, suggesting a large fragmentation of this second octupole phonon state in {sup 208}Pb. Upper limits for the B(E3) strength were determined for higher-lying, but unseen, 6{sup +} states ranging from 15{percent} of the harmonic value at 5.2 MeV to 100{percent} at 6.0 MeV. {copyright} {ital 1998} {ital The American Physical Society}

  16. Electromagnetically induced transparency from two-phonon processes in quadratically coupled membranes

    SciTech Connect

    Huang, Sumei; Agarwal, G. S.

    2011-02-15

    We describe how electromagnetically induced transparency can arise in quadratically coupled optomechanical systems. Due to quadratic coupling, the underlying optical process involves a two-phonon process in an optomechanical system, and this two-phonon process makes the mean displacement, which plays the role of atomic coherence in traditional electromagnetically induced transparency (EIT), zero. We show how the fluctuation in displacement can play a role similar to atomic coherence and can lead to EIT-like effects in quadratically coupled optomechanical systems. We show how such effects can be studied using the existing optomechanical systems.

  17. Covariant density functional theory with two-phonon coupling in nuclei

    SciTech Connect

    Ring, P.; Litvinova, E.; Tselyaev, V.

    2012-10-20

    A full description of excited states within the framework of density functional theory requires energy dependent self energies. We present a new class of many-body models. It allows a parameter free description of the fragmentation of nuclear states induced by mode coupling of two-quasiparticle and two-phonon configurations. The method is applied for an investigation of low-lying dipole excitations in Sn isotopes with large neutron excess.

  18. Hong–Ou–Mandel interference of two phonons in trapped ions

    NASA Astrophysics Data System (ADS)

    Toyoda, Kenji; Hiji, Ryoto; Noguchi, Atsushi; Urabe, Shinji

    2015-11-01

    The quantum statistics of bosons and fermions manifest themselves in the manner in which two indistinguishable particles interfere quantum mechanically. When two photons, which are bosonic particles, enter a beam-splitter with one photon in each input port, they bunch together at either of the two output ports. The corresponding disappearance of the coincidence count is the Hong–Ou–Mandel effect. Here we show the phonon counterpart of this effect in a system of trapped-ion phonons, which are collective excitations derived by quantizing vibrational motions that obey Bose–Einstein statistics. We realize a beam-splitter transformation of the phonons by employing the mutual Coulomb repulsion between ions, and perform a two-phonon quantum interference experiment using that transformation. We observe an almost perfect disappearance of the phonon coincidence between two ion sites, confirming that phonons can be considered indistinguishable bosonic particles. The two-particle interference demonstrated here is purely a quantum effect, without a classical counterpart, hence it should be possible to demonstrate the existence of entanglement on this basis. We attempt to generate an entangled state of phonons at the centre of the Hong–Ou–Mandel dip in the coincidence temporal profile, under the assumption that the entangled phonon state is successfully generated if the fidelity of the analysis pulses is taken into account adequately. Two-phonon interference, as demonstrated here, proves the bosonic nature of phonons in a trapped-ion system. It opens the way to establishing phonon modes as carriers of quantum information in their own right, and could have implications for the quantum simulation of bosonic particles and analogue quantum computation via boson sampling.

  19. Comparative study of the two-phonon Raman bands of silicene and graphene

    NASA Astrophysics Data System (ADS)

    Popov, Valentin N.; Lambin, Philippe

    2016-06-01

    We present a computational study of the two-phonon Raman spectra of silicene and graphene within a density-functional non-orthogonal tight-binding model. Due to the presence of linear bands close to the Fermi energy in the electronic structure of both structures, the Raman scattering by phonons is resonant. We find that the Raman spectra exhibit a crossover behavior for laser excitation close to the π-plasmon energy. This phenomenon is explained by the disappearance of certain paths for resonant Raman scattering and the appearance of other paths beyond this energy. Besides that, the electronic joint density of states (DOS) is divergent at this energy, which is reflected on the behavior of the Raman bands of the two structures in a qualitatively different way. Additionally, a number of Raman bands, originating from divergent phonon DOS at the M point and at points, inside the Brillouin zone, is also predicted. The calculated spectra for graphene are in excellent agreement with available experimental data. The obtained Raman bands can be used for structural characterization of silicene and graphene samples by Raman spectroscopy.

  20. Relaxation times of the two-phonon processes with spin-flip and spin-conserving in quantum dots

    SciTech Connect

    Wang, Zi-Wu; Liu, Lei; Li, Shu-Shen

    2014-04-07

    We perform a theoretical investigation on the two-phonon processes of the spin-flip and spin-conserving relaxation in quantum dots in the frame of the Huang-Rhys' lattice relaxation model. We find that the relaxation time of the spin-flip is two orders of magnitude longer than that of the spin-conserving, which is in agreement with previous experimental measurements. Moreover, the opposite variational trends of the relaxation time as a function of the energy separation for two-phonon processes are obtained in different temperature regime. The relaxation times display the oscillatory behaviors at the demarcation point with increasing magnetic field, where the energy separation matches the optical phonon energy and results in the optical phonon resonance. These results are useful in understanding the intraband levels' relaxation in quantum dots and could be helpful in designing photoelectric and spin-memory devices.

  1. First experimental evidence for two-phonon octupole-[gamma]-vibrational excitations in deformed nuclei

    SciTech Connect

    Kneissl, U.; Zilges, A.; Margraf, J.; Bauske, I.; von Brentano, P.; Friedrichs, H.; Heil, R.D.; Herzberg, R.; Pitz, H.H.; Schlitt, B.; Wesselborg, C. Institut fuer Kernphysik, Universitaet zu Koeln, 50937 Koeln Institut fuer Kernphysik, Universitaet Giessen, 35392 Giessen )

    1993-10-04

    Systematic nuclear resonance fluorescence experiments, including model independent parity determinations, provided evidence for enhanced [ital E]1 excitations near 2.5 MeV excitation energy in the deformed nuclei [sup 150]Nd, [sup 160]Gd, and [sup 162,164]Dy. The corresponding [ital J][sup [pi

  2. E0 decays of 0/sup +/ states in /sup 146/Gd: Search for two-phonon octupole excitations

    SciTech Connect

    Yates, S.W.; Mann, L.G.; Henry, E.A.; Decman, D.J.; Meyer, R.A.; Estep, R.J.; Julin, R.; Passoja, A.; Kantele, J.; Trzaska, W.; and others

    1987-11-01

    Conversion-electron measurements following the /sup 144/Sm(..cap alpha..,2n) and /sup 144/Sm( /sup 3/He,n) reactions reveal high-energy E0 transitions at 3020, 3485, and 3639 keV. The lowest of these deexcites the known two-neutron pairing vibrational state in /sup 146/Gd. It is not clear whether either of the others can be associated with the 0/sup +/ member of the 3/sup -/ x 3/sup -/ two-phonon octupole multiplet.

  3. Spectroscopy of excited states in 234U and search for a two-phonon state in 234U

    NASA Astrophysics Data System (ADS)

    Kotthaus, T.; Reiter, P.; Bruyneel, B.; Chatzistamatiou, M.; Eberth, J.; Gersch, G.; Hess, H.; Stefanescu, I.; Warr, N.; Weisshaar, D.; Wiens, A.; Morgan, T.; Lutter, R.; Schwerdtfeger, W.; Thirolf, P. G.; Bringel, P.; Hübel, H.; Neusser-Neffgen, A.

    2013-12-01

    Excited states of the isotope 234U were studied employing the one-neutron transfer reaction 235U(d,t)234U at a beam energy of 11 MeV. The reaction channel was selected by an identification of the outgoing tritons in a ΔE-E measurement. In coincidence with the tritons, γ rays were also detected with the highly efficient MINIBALL spectrometer. Based on the analysis of γγ coincidence data, the level scheme of 234U was extended. The 4+ state at an energy of 1886.7 keV provides evidence for a possible two-phonon γγ excitation, which is based on three γ transitions from this state to the γ vibrational band. However, this state is populated with a high cross section via the one-neutron transfer reaction, which is not expected for a highly collective excitation. The situation is compared to other known cases from the rare-earth region and to the neighboring isotone 232Th, which so far is the only known case for a two-phonon excitation in the actinide region.

  4. Calomel-made crystalline acousto-optical cell designed for an advanced regime of noncollinear two-phonon light scattering

    NASA Astrophysics Data System (ADS)

    Shcherbakov, Alexandre S.; Arellanes, Adan Omar

    2016-03-01

    We study the potentials of a wide-aperture crystalline calomel-made acousto-optical cell. Characterizing this cell is nontrivial due to the chosen regime based on an advanced noncollinear two-phonon light scattering. Recently revealed important features of this phenomenon are essentially exploited in the cell and are investigated in more detail. These features can be observed more easily and simply in tetragonal crystals, e.g., calomel, exhibiting specific acousto-optical nonlinearity caused by the acoustic waves of finite amplitude. This parametric nonlinearity manifests itself at low acoustic powers in calomel possessing linear acoustic attenuation. The formerly identified additional degree of freedom, unique to this regime, is exploited for designing the cell with an eye to doubling the resolution due to two-phonon processes. We clarify the role of varying the central acoustic frequency and acoustic attenuation using that degree of freedom. Then the efficiency of calomel is exploited to expand the cell's bandwidth with a cost of its efficiency. Proof-of-principle experiments confirm the developed approaches and illustrate their applicability to innovative techniques of optical spectrum analysis with the improved resolution. The achieved spectral resolution of 0.205 Å at 405 nm and the resolving power 19,800 are the best for acousto-optical spectrometers dedicated to space or airborne operations to date as far as we know.

  5. Weighted f-values, A-values, and line strengths for the E1 transitions among 3d 6, 3d 54s, and 3d 54p levels of Fe III

    NASA Astrophysics Data System (ADS)

    Deb, Narayan C.; Hibbert, Alan

    2009-03-01

    Weighted oscillator strengths, weighted radiative rates, and line strengths for all the E1 transitions between 285 fine-structure levels belonging to the 3d 6, 3d 54s, and 3d 54p configurations of Fe III are presented, in ascending order of wavelength. Calculations have been undertaken using the general configuration interaction (CI) code CIV3. The large configuration set is constructed by allowing single and double replacements from any of 3d 6, 3d 54s, 3d 54p, and 3d 54d configurations to nl orbitals with n⩽5,l⩽3 as well as 6p. Additional selective promotions from 3s and 3p subshells are also included in the CI expansions to incorporate the important correlation effects in the n=3 shell. Results of some strong transitions between levels of 3d 6, 3d 54s, and 3d 54p configurations are also presented and compared with other available calculations. It is found that large disagreements occur in many transitions among the existing calculations.

  6. The E1 proteins

    SciTech Connect

    Bergvall, Monika; Melendy, Thomas; Archambault, Jacques

    2013-10-15

    E1, an ATP-dependent DNA helicase, is the only enzyme encoded by papillomaviruses (PVs). It is essential for replication and amplification of the viral episome in the nucleus of infected cells. To do so, E1 assembles into a double-hexamer at the viral origin, unwinds DNA at the origin and ahead of the replication fork and interacts with cellular DNA replication factors. Biochemical and structural studies have revealed the assembly pathway of E1 at the origin and how the enzyme unwinds DNA using a spiral escalator mechanism. E1 is tightly regulated in vivo, in particular by post-translational modifications that restrict its accumulation in the nucleus. Here we review how different functional domains of E1 orchestrate viral DNA replication, with an emphasis on their interactions with substrate DNA, host DNA replication factors and modifying enzymes. These studies have made E1 one of the best characterized helicases and provided unique insights on how PVs usurp different host-cell machineries to replicate and amplify their genome in a tightly controlled manner. - Highlights: • The papillomavirus E1 helicase orchestrates replication of the viral DNA genome. • E1 assembles into a double-hexamer at the viral origin with the help of E2. • E1 interacts with cellular DNA replication factors. • E1 unwinds DNA using a spiral escalator mechanism. • Nuclear accumulation of E1 is regulated by post-translational modifications.

  7. Oscillator Strengths and Predissociation Rates for Rydberg Transitions in 12C16O, 13C16O, and 13C18O Involving the E 1Π, B 1Σ+, and W 1Π States

    NASA Astrophysics Data System (ADS)

    Eidelsberg, M.; Sheffer, Y.; Federman, S. R.; Lemaire, J. L.; Fillion, J. H.; Rostas, F.; Ruiz, J.

    2006-08-01

    One of the processes controlling the interstellar CO abundance and the ratio of its isotopologues is photodissociation. Accurate oscillator strengths and predissociation rates for Rydberg transitions are needed for modeling this process. We present results on absorption from the E 1Π-X 1Σ+ (1-0) and B 1Σ+-X 1Σ+ (6-0) bands at 1051 and 1002 Å, respectively, and the vibrational progression W 1Π-X 1Σ+ (v'-0) bands with v'=0-3 at 972, 956, 941, and 925 Å, respectively. The corresponding spectra were acquired at the high resolution (R~30,000) SU5 beam line at the Super ACO Synchrotron in Orsay, France. Spectra were obtained for the 12C16O, 13C 16O, and 13C18O isotopologues. These represent the most complete set of measurements available. Comparison is made with earlier results, both empirical and theoretical. While earlier determinations of oscillator strengths based on absorption from synchrotron radiation tend to be somewhat smaller than ours, the suite of measurements from a variety of techniques agree for the most part, considering the mutual uncertainties. For the bands studied here, their relative weakness, or their significant line widths arising from predissociation, minimizes potential problems from large optical depths at line center in absorption measurements. Predissociating line widths could generally be extracted from the spectra thanks to the profile simulations used in the analysis. In many cases, these simulations allowed us to consider e and f parity levels separately and to determine the dependence of the width on rotational quantum number, J. Our results are consistent with earlier determinations, especially the widths inferred from laser experiments.

  8. Direct and two-phonon Orbach-Aminov type spin-lattice relaxation in molecular magnet V15

    NASA Astrophysics Data System (ADS)

    Tarantul, Alex; Tsukerblat, Boris

    2011-10-01

    In this article we propose a model of spin-phonon relaxation in K6[VIV 15As6O42(H2O)]-8H2O, the so called V15 cluster exhibiting the unique layered magnetic structure. The work is motivated by the recent observation of the Rabi oscillation [1] in this system and aimed to elucidate the role of spin-phonon interaction as a source of decoherence. The spin-phonon coupling is assumed to appear as a result of the modulation of the isotropic and antisymmetric (Dzyaloshinsky-Moriya) exchange interactions in the central triangular layer of vanadium ions by the acoustic lattice vibrations. The relaxation rates are estimated within the Debye model for the lattice vibrations. Within the pseudo-angular momentum representation the selection rules for the direct (one-phonon) transitions between Zeeman levels are derived and a special role of the antisymmetric exchange is underlined. The probabilities of the two-phonon Orbach-Aminov type processes are evaluated as well, while the Raman type relaxation is shown to have a negligible importance at low temperatures at which the Rabi oscillations have been detected.

  9. Two-Phonon Absorption

    ERIC Educational Resources Information Center

    Hamilton, M. W.

    2007-01-01

    A nonlinear aspect of the acousto-optic interaction that is analogous to multi-photon absorption is discussed. An experiment is described in which the second-order acousto-optically scattered intensity is measured and found to scale with the square of the acoustic intensity. This experiment using a commercially available acousto-optic modulator is…

  10. Two-Phonon Absorption

    ERIC Educational Resources Information Center

    Hamilton, M. W.

    2007-01-01

    A nonlinear aspect of the acousto-optic interaction that is analogous to multi-photon absorption is discussed. An experiment is described in which the second-order acousto-optically scattered intensity is measured and found to scale with the square of the acoustic intensity. This experiment using a commercially available acousto-optic modulator is

  11. Nature of One- and Two-Phonon Mixed Symmetry States in 92Zr and 94Mo from High-Resolution Electron and Proton Scattering

    SciTech Connect

    Neumann-Cosel, P. von; Burda, O.; Kuhar, M.; Lenhardt, A.; Ponomarev, V. Yu.; Richter, A.; Wambach, J.; Botha, N. T.; Fearick, R. W.; Carter, J.; Sideras-Haddad, E.; Foertsch, S. V.; Neveling, R.; Smit, F. D.; Fransen, C.; Fujita, H.; Pietralla, N.

    2006-03-13

    High-resolution inelastic electron (performed at the S-DALINAC) and proton (performed at iThemba LABS) scattering experiments on 92Zr and 94Mo with emphasis on E2 transitions are presented The measured form factors and angular distributions provide a measure for the F-spin purity, respectively the isovector nature, of the proposed one-phonon mixed symmetry states and furthermore provide a sensitive test of a possible two-phonon character of excited 2+ states.

  12. Search for one- and two-phonon octupole vibrational states in the spherical nuclei near 132Sn

    NASA Astrophysics Data System (ADS)

    Hwang, J. K.; Hamilton, J. H.; Ramayya, A. V.; Luo, Y. X.

    2013-10-01

    Excited high spin states in 135I, 136Xe, 137Cs, 138Ba, 139La, 140Ce and 142Nd with N = 82 are reorganized and interpreted in a different way to find one- phonon octupole vibrational (POV) bands. Two nearly identical (similar) bands with ΔI = 3 are found in these nuclei. From the presence of two nearly identical excited bands with ΔI = 3 in these nuclei, one-POV bands are proposed. Also, high spin states of 134Sb, 134,135Te, 135,136I, 137Xe and 139Ba near 132Sn are reanalyzed in order to search for one- and two-POV states. New spins and parities are tentatively assigned to the 2203.9 keV state in 137Xe and the 1976.6 and 2091.7 keV states in 139Ba from the state energy plots of the N = 82 and 83 nuclei. High spin states of 134Sb, 134,135Te, 135,136I, 137Xe and 139Ba connected by E1, E3 /M2 and E3 transitions are proposed, for the first time, as zero-, one- and two-POV states. One- and two-POV states in 134Sb and 135Te are built on a 7- (πg7/2 ν f7/2) state and a 19 /2- (νf7/2 ⊗ 61+)state, respectively. One-POV states built on the 19 /2- (ν f7/2 ⊗ 61+)and the 21 /2- (νh9/2 ⊗ 62+)states coexist in 137Xe. Then, one- and two-POV states in 139Ba are built only on the 21 /2- (νh9/2 ⊗ 62+)state. One- and two-POV states in 134Te are built on the 62+state with some mixing with the 61+state.

  13. Influence of neutron surface on E1 resonance properties

    NASA Astrophysics Data System (ADS)

    Goncharova, N. G.; Tretyakova, T. Yu.; Fedorov, N. A.

    2016-01-01

    The E1 strength distributions in even-even Si isotopes were calculated in the "particle-core coupling" version of the shell model taking into account the fragmentation of the hole configuration among the states of the daughter nuclei. The comparison of calculated strength distributions in different isotopes of the same element shows the peculiarities of a neutron surface influence on the E1 resonance fragmentation.

  14. Weighted f-values, A-values, and line strengths for the E1 transitions among 3d{sup 6}, 3d{sup 5}4s, and 3d{sup 5}4p levels of Fe III

    SciTech Connect

    Deb, Narayan C.; Hibbert, Alan

    2009-03-15

    Weighted oscillator strengths, weighted radiative rates, and line strengths for all the E1 transitions between 285 fine-structure levels belonging to the 3d{sup 6}, 3d{sup 5}4s, and 3d{sup 5}4p configurations of Fe III are presented, in ascending order of wavelength. Calculations have been undertaken using the general configuration interaction (CI) code CIV3. The large configuration set is constructed by allowing single and double replacements from any of 3d{sup 6}, 3d{sup 5}4s, 3d{sup 5}4p, and 3d{sup 5}4d configurations to nl orbitals with n{<=}5,l{<=}3 as well as 6p. Additional selective promotions from 3s and 3p subshells are also included in the CI expansions to incorporate the important correlation effects in the n=3 shell. Results of some strong transitions between levels of 3d{sup 6}, 3d{sup 5}4s, and 3d{sup 5}4p configurations are also presented and compared with other available calculations. It is found that large disagreements occur in many transitions among the existing calculations.

  15. Gene coding for the E1 endoglucanase

    DOEpatents

    Thomas, S.R.; Laymon, R.A.; Himmel, M.E.

    1996-07-16

    The gene encoding Acidothermus cellulolyticus E1 endoglucanase is cloned and expressed in heterologous microorganisms. A new modified E1 endoglucanase enzyme is produced along with variants of the gene and enzyme. The E1 endoglucanase is useful for hydrolyzing cellulose to sugars for simultaneous or later fermentation into alcohol. 6 figs.

  16. Gene coding for the E1 endoglucanase

    DOEpatents

    Thomas, Steven R.; Laymon, Robert A.; Himmel, Michael E.

    1996-01-01

    The gene encoding Acidothermus cellulolyticus E1 endoglucanase is cloned and expressed in heterologous microorganisms. A new modified E1 endoglucanase enzyme is produced along with variants of the gene and enzyme. The E1 endoglucanase is useful for hydrolyzing cellulose to sugars for simultaneous or later fermentation into alcohol.

  17. Adenovirus E1B 19-kilodalton protein overcomes the cytotoxicity of E1A proteins.

    PubMed Central

    White, E; Cipriani, R; Sabbatini, P; Denton, A

    1991-01-01

    Infection with adenovirus mutants carrying either point mutations or deletions in the coding region for the 19-kDa E1B gene product (19K protein) causes degradation of host cell and viral DNAs (deg phenotype) and enhanced cytopathic effect (cyt phenotype). Therefore, one function of the E1B 19K protein is to protect nuclear DNA integrity and preserve cytoplasmic architecture during productive adenovirus infection. When placed in the background of a virus incapable of expressing a functional E1A gene product, however, E1B 19K gene mutations do not result in the appearance of the cyt and deg phenotypes. This demonstrated that expression of the E1A proteins was responsible for inducing the appearance of the cyt and deg phenotypes. By constructing a panel of viruses possessing E1A mutations spanning each of the three E1A conserved regions in conjunction with E1B 19K gene mutations, we mapped the induction of the cyt and deg phenotypes to the amino-terminal region of E1A. Viruses that fail to express conserved region 3 (amino acids 140 to 185) and/or 2, (amino acids 121 to 185) or nonconserved sequences between conserved regions 2 and 1 of E1A (amino acids 86 to 120) were still capable of inducing cyt and deg. This indicated that activities associated with these regions, such as transactivation and binding to the product of the retinoblastoma susceptibility gene, were dispensable for induction of E1A-dependent cytotoxic effects. In contrast, deletion of sequences in the amino terminus of E1A (amino acids 22 to 107) resulted in extragenic suppression of the cyt and deg phenotypes. Therefore, a function affected by deletion of amino acids 22 to 86 of E1A is responsible for exerting cytotoxic effects in virally infected cells. Furthermore, transient high-level expression of the E1A region using a cytomegalovirus promoter plasmid expression vector was sufficient to induce the cyt and deg phenotypes, demonstrating that E1A expression alone is sufficient to exert these cytotoxic effects and that other viral gene products are not involved. Finally, placing E1A expression under the control of a strong promoter did not alter the requirement for E1B in the transformation of primary cells. One possibility is that the E1B 19K protein is required to overcome the cytotoxic effects of E1A protein expression and thereby enable primary cells to become transformed. Images PMID:1851867

  18. Energy-dependence of skin-mode fraction in E1 excitations of neutron-rich nuclei

    NASA Astrophysics Data System (ADS)

    Nakada, H.; Inakura, T.; Sawai, H.

    2015-05-01

    We have extensively investigated characters of the low-energy E1 strengths in N > Z nuclei, by analyzing the transition densities obtained by the HF+RPA calculations with several effective interactions. Crossover behavior has been confirmed, from the skin mode at low energy to the pn mode at higher energy. Decomposing the E1 strengths into the skin-mode, pn-mode and interference fractions, we show that the ratio of the skin-mode strength to the full strength may be regarded as a generic function of the excitation energy, insensitive to nuclides and effective interactions, particularly beyond Ni.

  19. Genetic polymorphism in CYP2E1: Population distribution of CYP2E1 activity.

    PubMed

    Neafsey, Pat; Ginsberg, Gary; Hattis, Dale; Johns, Douglas O; Guyton, Kathryn Z; Sonawane, Babasaheb

    2009-01-01

    Cytochrome P-450 2E1 (CYP2E1) is a key enzyme in the metabolic activation of a variety of toxicants including nitrosamines, benzene, vinyl chloride, and halogenated solvents such as trichloroethylene. CYP2E1 is also one of the enzymes that metabolizes ethanol to acetaldehyde, and is induced by recent ethanol ingestion. There is evidence that interindividual variability in the expression and functional activity of this cytochrome (CYP) may be considerable. Genetic polymorphisms in CYP2E1 were identified and linked to altered susceptibility to hepatic cirrhosis induced by ethanol and esophageal and other cancers in some epidemiological studies. Therefore, it is important to evaluate how such polymorphisms affect CYP2E1 function and whether it is possible to construct a population distribution of CYP2E1 activity based upon the known effects of these polymorphisms and their frequency in the population. This analysis is part of the genetic polymorphism database project described in the lead article in this series and followed the approach described in that article (Ginsberg et al., 2009, this issue). Review of the literature found that there are a variety of CYP2E1 variant alleles but the functional significance of these variants is still unclear. Some, but not all, studies suggest that several upstream 5' flanking mutations affect gene expression and response to inducers such as ethanol or obesity. None of the coding-region variants consistently affects enzyme function. Part of the reason for conflicting evidence regarding genotype effect on phenotype may be due to the wide variety of exposures such as ethanol or dietary factors and physiological factors including body weight or diabetes that modulate CYP2E1 expression. In conclusion, evidence is too limited to support the development of a population distribution of CYP2E1 enzyme activity based upon genotypes. Health risk assessments may best rely upon data reporting interindividual variability in CYP2E1 function for input into physiologically based pharmacokinetic (PBPK) models involving CYP2E1 substrates. PMID:20183527

  20. Strength Testing.

    ERIC Educational Resources Information Center

    Londeree, Ben R.

    1981-01-01

    Postural deviations resulting from strength and flexibility imbalances include swayback, scoliosis, and rounded shoulders. Screening tests are one method for identifying strength problems. Tests for the evaluation of postural problems are described, and exercises are presented for the strengthening of muscles. (JN)

  1. Strength Training

    MedlinePlus

    ... great way to improve strength, endurance, and muscle tone. But remember to start slowly, use proper form, ... week will really pay off — besides better muscle tone and definition, you may find that you have ...

  2. Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication.

    PubMed

    Krömmelbein, Natascha; Wiebusch, Lüder; Schiedner, Gudrun; Büscher, Nicole; Sauer, Caroline; Florin, Luise; Sehn, Elisabeth; Wolfrum, Uwe; Plachter, Bodo

    2016-01-01

    The human cytomegalovirus (HCMV) replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP) is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production. PMID:26848680

  3. Adenovirus E1A/E1B Transformed Amniotic Fluid Cells Support Human Cytomegalovirus Replication

    PubMed Central

    Krömmelbein, Natascha; Wiebusch, Lüder; Schiedner, Gudrun; Büscher, Nicole; Sauer, Caroline; Florin, Luise; Sehn, Elisabeth; Wolfrum, Uwe; Plachter, Bodo

    2016-01-01

    The human cytomegalovirus (HCMV) replicates to high titers in primary human fibroblast cell cultures. A variety of primary human cells and some tumor-derived cell lines do also support permissive HCMV replication, yet at low levels. Cell lines established by transfection of the transforming functions of adenoviruses have been notoriously resistant to HCMV replication and progeny production. Here, we provide first-time evidence that a permanent cell line immortalized by adenovirus type 5 E1A and E1B (CAP) is supporting the full HCMV replication cycle and is releasing infectious progeny. The CAP cell line had previously been established from amniotic fluid cells which were likely derived from membranes of the developing fetus. These cells can be grown under serum-free conditions. HCMV efficiently penetrated CAP cells, expressed its immediate-early proteins and dispersed restrictive PML-bodies. Viral DNA replication was initiated and viral progeny became detectable by electron microscopy in CAP cells. Furthermore, infectious virus was released from CAP cells, yet to lower levels compared to fibroblasts. Subviral dense bodies were also secreted from CAP cells. The results show that E1A/E1B expression in transformed cells is not generally repressive to HCMV replication and that CAP cells may be a good substrate for dense body based vaccine production. PMID:26848680

  4. Many trained at E1 Salvador center.

    PubMed

    1970-02-01

    The Family Planning Association of E1 Salvador is conducting regular week-long courses in family planning for doctors, nurses, social workers and other interested people in El Salvador and the surrounding countries of Guatemala, Honduras, Nicaragua, Costa Rica and Panama. Twelve regional courses on population dynamics, the physiology of reproduction and family planning were held in the academic year 1968-69 at the Faculty of Medicine at the University of El Salvador. Because the demand for places far exceeds the capacity of the Association's training unit, careful selection of candidates is made through consultation with the national family planning associations. Most of those trained are doctors who are being specially equipped to carry out family planning work either as part of government maternal and child health centres or in Association clinics. Many places are also found for nurses and some social workers also attend in order to build up teams for effective clinic management. A few journalists and teachers have been among the trainees as well as groups of religious leaders including Roman Catholics. Lecturers are mainly university personnel drawn from a wide range of disciplines in order to relate family planning not only to health and medicine but also to socio-economic aspects and community welfare. The El Salvador Association, an IPPF member, gets financial support from the Population Council for the training programme but hopes eventually that responsibility for continuing the courses will be taken over by the Government, probably through the Ministry of Education. Another pace-setting activity of the Association has been its close contacts with industry, particulary through the efforts of its President, Mr. Lucio Burgos, General Manager of the El Salvador Power Company, who has gained the interest and support of business and union leaders. Not only do these groups help the work of the Association through fund-raising and public relations activities, but they co-operate with the Association in introducing family planning to their workers through group meetings and talks. PMID:12275386

  5. 26 CFR 1.167(e)-1 - Change in method.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... to December 30, 2003 (§ 1.167(e)-1 as contained in 26 CFR part 1 edition revised as of April 1, 2003). ... 26 Internal Revenue 2 2010-04-01 2010-04-01 false Change in method. 1.167(e)-1 Section 1.167(e)-1...) INCOME TAXES (CONTINUED) Itemized Deductions for Individuals and Corporations § 1.167(e)-1 Change...

  6. 26 CFR 31.3231(e)-1 - Compensation.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 15 2011-04-01 2011-04-01 false Compensation. 31.3231(e)-1 Section 31.3231(e)-1... Retirement Tax Act (Chapter 22, Internal Revenue Code of 1954) General Provisions § 31.3231(e)-1 Compensation. (a) Definition—(1) The term compensation has the same meaning as the term wages in section...

  7. 26 CFR 31.3231(e)-1 - Compensation.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false Compensation. 31.3231(e)-1 Section 31.3231(e)-1... Retirement Tax Act (Chapter 22, Internal Revenue Code of 1954) General Provisions § 31.3231(e)-1 Compensation. (a) Definition—(1) The term compensation has the same meaning as the term wages in section...

  8. 26 CFR 31.3231(e)-1 - Compensation.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 15 2012-04-01 2012-04-01 false Compensation. 31.3231(e)-1 Section 31.3231(e)-1... Retirement Tax Act (Chapter 22, Internal Revenue Code of 1954) General Provisions § 31.3231(e)-1 Compensation. (a) Definition—(1) The term compensation has the same meaning as the term wages in section...

  9. 26 CFR 31.3231(e)-1 - Compensation.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 15 2014-04-01 2014-04-01 false Compensation. 31.3231(e)-1 Section 31.3231(e)-1... Retirement Tax Act (Chapter 22, Internal Revenue Code of 1954) General Provisions § 31.3231(e)-1 Compensation. (a) Definition—(1) The term compensation has the same meaning as the term wages in section...

  10. 26 CFR 31.3231(e)-1 - Compensation.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Compensation. 31.3231(e)-1 Section 31.3231(e)-1... Retirement Tax Act (Chapter 22, Internal Revenue Code of 1954) General Provisions § 31.3231(e)-1 Compensation. (a) Definition—(1) The term compensation has the same meaning as the term wages in section...

  11. 26 CFR 53.4941(e)-1 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 17 2012-04-01 2012-04-01 false Definitions. 53.4941(e)-1 Section 53.4941(e)-1... TAXES (CONTINUED) FOUNDATION AND SIMILAR EXCISE TAXES Taxes on Self-Dealing § 53.4941(e)-1 Definitions... disqualified person a binding option on June 15, 1971, to purchase property owned by the foundation at any...

  12. 26 CFR 53.4941(e)-1 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 17 2011-04-01 2011-04-01 false Definitions. 53.4941(e)-1 Section 53.4941(e)-1... TAXES (CONTINUED) FOUNDATION AND SIMILAR EXCISE TAXES Taxes on Self-Dealing § 53.4941(e)-1 Definitions... disqualified person a binding option on June 15, 1971, to purchase property owned by the foundation at any...

  13. 26 CFR 53.4941(e)-1 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 17 2013-04-01 2013-04-01 false Definitions. 53.4941(e)-1 Section 53.4941(e)-1... TAXES (CONTINUED) FOUNDATION AND SIMILAR EXCISE TAXES Taxes on Self-Dealing § 53.4941(e)-1 Definitions... disqualified person a binding option on June 15, 1971, to purchase property owned by the foundation at any...

  14. 26 CFR 53.4941(e)-1 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 17 2014-04-01 2014-04-01 false Definitions. 53.4941(e)-1 Section 53.4941(e)-1... TAXES (CONTINUED) FOUNDATION AND SIMILAR EXCISE TAXES Taxes on Self-Dealing § 53.4941(e)-1 Definitions... disqualified person a binding option on June 15, 1971, to purchase property owned by the foundation at any...

  15. 26 CFR 53.4941(e)-1 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 17 2010-04-01 2010-04-01 false Definitions. 53.4941(e)-1 Section 53.4941(e)-1... TAXES (CONTINUED) FOUNDATION AND SIMILAR EXCISE TAXES Taxes on Self-Dealing § 53.4941(e)-1 Definitions... been accepted, or because the deficiency is paid, the date of filing of the waiver or the date of...

  16. CYP2E1 and Oxidative Liver Injury by Alcohol

    PubMed Central

    Lu, Yongke; Cederbaum, Arthur I.

    2008-01-01

    Ethanol-induced oxidative stress appears to play a major role in mechanisms by which ethanol causes liver injury. Many pathways have been suggested to contribute to the ability of ethanol to induce a state of oxidative stress. One central pathway appears to be the induction of cytochrome P450 2E1 (CYP2E1) by ethanol. CYP2E1 metabolizes and activates many toxicological substrates, including ethanol, to more reactive, toxic products. Levels of CYP2E1 are elevated under a variety of physiological and pathophysiological conditions, and after acute and chronic alcohol treatment. CYP2E1 is also an effective generator of reactive oxygen species such as the superoxide anion radical and hydrogen peroxide, and in the presence of iron catalysts, produces powerful oxidants such as the hydroxyl radical. This Review Article summarizes some of the biochemical and toxicological properties of CYP2E1, and briefly describes the use of cell lines developed to constitutively express CYP2E1 in assessing the actions of CYP2E1. Possible therapeutic implications for treatment of alcoholic liver injury by inhibition of CYP2E1 or CYP2E1-dependent oxidative stress will be discussed, followed by some future directions which may help to understand the actions of CYP2E1 and its role in alcoholic liver injury. PMID:18078827

  17. Activation of adenovirus 5 E1A transcription by region E1B in transformed primary rat cells.

    PubMed Central

    Jochemsen, A G; Peltenburg, L T; te Pas, M F; de Wit, C M; Bos, J L; van der Eb, A J

    1987-01-01

    The human adenovirus 5 E1A region can immortalize primary cultures of baby rat kidney cells, but requires the presence of the E1B region for complete oncogenic transformation. One of the effects of the E1B region in the transformation process is the activation of E1A expression. We have investigated the mechanism of this stimulation of E1A expression using nuclear run-on assays with nuclei from Ad5 E1A- and Ad5 E1-transformed cells. It was found that E1B enhances E1A at the level of transcription-initiation. This activation is mainly observed when the E1A and E1B regions are integrated simultaneously into the cellular genome and only minimally when these genes are integrated separately, strongly suggesting that a close physical linkage of these regions is essential for the observed effect. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. PMID:2962857

  18. Study of M1 and E1 excitations by high-resolution proton inelastic scattering measurement at forward angles

    SciTech Connect

    Tamii, A.; Adachi, T.; Hatanaka, K.; Hashimoto, H.; Kaneda, T.; Matsubara, H.; Okamura, H.; Sakemi, Y.; Shimizu, Y.; Tameshige, Y.; Yosoi, M.; Carter, J.; Dozono, M.; Fujita, H.; Fujita, Y.; Itoh, M.; Kawabata, T.; Nakanishi, K.; Sasamoto, Y.; Neumann-Cosel, P. von

    2007-06-13

    Experimental technique for measuring proton inelastic scattering with high-resolution at 295 MeV and at forward angles including zero degrees is described. The method is useful for extracting spin part of the M1 strength via nuclear excitation as well as E1 strength via Coulomb excitation. An excitation energy resolution of 20 keV, good scattering angle resolution, and low background condition have been achieved. The experimental technique was applied for several sd and pf shell nuclei.

  19. The nuclear receptor NR2E1/TLX controls senescence

    PubMed Central

    Krusche, Benjamin; Pemberton, Helen; Alonso, Marta M.; Chandler, Hollie; Brookes, Sharon; Parrinello, Simona; Peters, Gordon; Gil, Jesús

    2014-01-01

    The nuclear receptor NR2E1 (also known as TLX or tailless) controls the self-renewal of neural stem cells (NSCs) and has been implied as an oncogene which initiates brain tumours including glioblastomas. Despite NR2E1 regulating targets like p21CIP1 or PTEN we still lack a full explanation for its role in NSC self-renewal and tumorigenesis. We know that Polycomb repressive complexes (PRC) also control stem cell self-renewal and tumorigenesis, but so far, no formal connection has been established between NR2E1 and PRCs. In a screen for transcription factors regulating the expression of the Polycomb protein CBX7, we identified NR2E1 as one of its more prominent regulators. NR2E1 binds at the CBX7 promoter, inducing its expression. Notably CBX7 represses NR2E1 as part of a regulatory loop. Ectopic NR2E1 expression inhibits cellular senescence, extending cellular lifespan in fibroblasts via CBX7-mediated regulation of p16INK4a and direct repression of p21CIP1. In addition NR2E1 expression also counteracts oncogene-induced senescence (OIS). The importance of NR2E1 to restrain senescence is highlighted through the process of knocking down its expression, which causes premature senescence in human fibroblasts and epithelial cells. We also confirmed that NR2E1 regulates CBX7 and restrains senescence in NSCs. Finally, we observed that the expression of NR2E1 directly correlates with that of CBX7 in human glioblastoma multiforme. Overall we identified control of senescence and regulation of Polycomb action as two possible mechanisms that can join those so far invoked to explain the role of NR2E1 in control of NSC self-renewal and cancer. PMID:25328137

  20. Inhibition of Corneal Inflammation by the Resolvin E1

    PubMed Central

    Lee, Ji-Eun; Sun, Yan; Gjorstrup, Per; Pearlman, Eric

    2015-01-01

    Purpose. To investigate the role of the lipid mediator, resolvin E1 (RvE1), in corneal inflammation. Methods. The effect of RvE1 on stimulated human corneal epithelial cells (HCECs) and neutrophils, and mouse macrophage was assessed. C57BL/6 mouse corneas were abraded and treated with RvE1 either before or after stimulation with lipopolysaccharide (LPS) and antibiotic-killed Pseudomonas aeruginosa and Staphylococcus aureus. The levels of CXC chemokines in the cornea were quantified, and the presence of neutrophils in corneal infiltrates was detected by immunohistochemistry and by in vivo confocal microscopy. The effect of RvE1 on apoptosis in the corneal epithelium was assessed using the TUNEL assay. Results. RvE1 significantly inhibited cytokine production in HCECs and neutrophils, and mouse macrophages and cornea. The development of corneal infiltrates, specifically neutrophils, in response to stimulation with LPS, P. aeruginosa, and S. aureus was also significantly reduced. There was no apoptotic effect of RvE1 on mouse corneal epithelial cells. Conclusions. RvE1 inhibits corneal inflammation induced by LPS, Gram negative (P. aeruginosa) and Gram positive (S. aureus) bacteria. These findings indicate that RvE1 as a potential anti-inflammatory therapy for patients with corneal inflammation and also, when given together with antibiotics, for bacterial keratitis. PMID:25758817

  1. CYP2E1 autoantibodies in liver diseases

    PubMed Central

    Sutti, Salvatore; Rigamonti, Cristina; Vidali, Matteo; Albano, Emanuele

    2014-01-01

    Autoimmune reactions involving cytochrome P4502E1 (CYP2E1) are a feature of idiosyncratic liver injury induced by halogenated hydrocarbons and isoniazid, but are also detectable in about one third of the patients with advanced alcoholic liver disease (ALD) and chronic hepatitis C (CHC). In these latter the presence of anti-CYP2E1 auto-antibodies is an independent predictor of extensive necro-inflammation and fibrosis and worsens the recurrence of hepatitis following liver transplantation, indicating that CYP2E1-directed autoimmunity can contribute to hepatic injury. The molecular characterization of the antigens recognized by anti-CYP2E1 auto-antibodies in ALD and CHC has shown that the targeted conformational epitopes are located in close proximity on the molecular surface. Furthermore, these epitopes can be recognized on CYP2E1 expressed on hepatocyte plasma membranes where they can trigger antibody-mediated cytotoxicity. This does not exclude that T cell-mediated responses against CYP2E1 might also be involved in causing hepatocyte damage. CYP2E1 structural modifications by reactive metabolites and molecular mimicry represent important factors in the breaking of self-tolerance against CYP2E1 in, respectively, ALD and CHC. However, genetic or acquired interferences with the mechanisms controlling the homeostasis of the immune system are also likely to contribute. More studies are needed to better characterize the impact of anti-CYP2E1 autoimmunity in liver diseases particularly in relation to the fact that common metabolic alterations such as obesity and diabetes stimulates hepatic CYP2E1 expression. PMID:25462068

  2. Chemical characteristics for optimizing CYP2E1 inhibition.

    PubMed

    van de Wier, B; Balk, J M; Bast, A; Koek, G H; Haenen, G R M M

    2015-12-01

    Cytochrome P450 2E1 (CYP2E1) expression and activity in the liver is associated with the degree of liver damage in patients with alcoholic steatohepatitis (ASH) as well as non-alcoholic steatohepatitis (NASH). CYP2E1 is known to generate reactive oxygen species, which leads to oxidative stress, one of the hallmarks of both diseases. Apart from ROS, toxic metabolites can be formed by CYP2E1 metabolism, further potentiating liver injury. Therefore, CYP2E1 is implicated in the pathogenesis of ASH and NASH. The aim of this study was to determine the chemical characteristics of compounds that are important to inhibit CYP2E1. To this end, structurally related analogs that differed in their lipophilic, steric and electronic properties were tested. In addition, homologues series of aliphatic primary alcohols, secondary alcohols, aldehydes, ketones and carboxylic acids were tested. It was found that inhibition of the CYP2E1 activity is primarily governed by lipophilicity. The optimal log D7.4 (octanol/water distribution coefficient at pH 7.4) value for inhibition of CYP2E1 was approximately 2.4. In the carboxylic acids series the interaction of the carboxylate group with polar residues lining the CYP2E1 active site also has to be considered. This study sketches the basic prerequisites in the search for inhibitors of CYP2E1, which would strengthen our therapeutic armamentarium against CYP2E1 associated diseases, such as ASH and NASH. PMID:26428356

  3. CYP2E1 autoantibodies in liver diseases.

    PubMed

    Sutti, Salvatore; Rigamonti, Cristina; Vidali, Matteo; Albano, Emanuele

    2014-01-01

    Autoimmune reactions involving cytochrome P4502E1 (CYP2E1) are a feature of idiosyncratic liver injury induced by halogenated hydrocarbons and isoniazid, but are also detectable in about one third of the patients with advanced alcoholic liver disease (ALD) and chronic hepatitis C (CHC). In these latter the presence of anti-CYP2E1 auto-antibodies is an independent predictor of extensive necro-inflammation and fibrosis and worsens the recurrence of hepatitis following liver transplantation, indicating that CYP2E1-directed autoimmunity can contribute to hepatic injury. The molecular characterization of the antigens recognized by anti-CYP2E1 auto-antibodies in ALD and CHC has shown that the targeted conformational epitopes are located in close proximity on the molecular surface. Furthermore, these epitopes can be recognized on CYP2E1 expressed on hepatocyte plasma membranes where they can trigger antibody-mediated cytotoxicity. This does not exclude that T cell-mediated responses against CYP2E1 might also be involved in causing hepatocyte damage. CYP2E1 structural modifications by reactive metabolites and molecular mimicry represent important factors in the breaking of self-tolerance against CYP2E1 in, respectively, ALD and CHC. However, genetic or acquired interferences with the mechanisms controlling the homeostasis of the immune system are also likely to contribute. More studies are needed to better characterize the impact of anti-CYP2E1 autoimmunity in liver diseases particularly in relation to the fact that common metabolic alterations such as obesity and diabetes stimulates hepatic CYP2E1 expression. PMID:25462068

  4. New Insights into FoxE1 Functions: Identification of Direct FoxE1 Targets in Thyroid Cells

    PubMed Central

    Fernández, Lara P.; López-Márquez, Arístides; Martínez, Ángel M.; Gómez-López, Gonzalo; Santisteban, Pilar

    2013-01-01

    Background FoxE1 is a thyroid-specific forkhead transcription factor essential for thyroid gland development, as well as for the maintenance of the thyroid differentiated state in adults. FoxE1 recognizes and binds to a short DNA sequence present in thyroglobulin (Tg) and thyroperoxidase (Tpo) promoters, but FoxE1 binding to regulatory regions other than Tg and Tpo promoters remains almost unexplored. Improving knowledge of the regulatory functions of FoxE1 is necessary to clarify its role in endocrine syndromes and cancer susceptibility. Methodology/Principal Finding In order to further investigate downstream FoxE1 targets, we performed a genome-wide expression screening after knocking-down FoxE1 and obtained new insights into FoxE1 transcriptional networks in thyroid follicular cells. After validation, we confirmed Adamts9, Cdh1, Duox2 and S100a4 as upregulated genes and Casp4, Creld2, Dusp5, Etv5, Hsp5a, Nr4a2 and Tm4sf1 as downregulated genes when FoxE1 was silenced. In promoter regions of putative FoxE1-regulated genes and also in the promoters of the classical thyroid genes Nis, Pax8 and Titf1, we performed an in silico search of the FoxE1 binding motif that was in close proximity to the NF1/CTF binding sequence, as previously described for other forkhead factors. Using chromatin immunoprecipitation we detected specific in vivo FoxE1 binding to novel regulatory regions in two relevant thyroid genes, Nis and Duox2. Moreover, we demonstrated simultaneous binding of FoxE1 and NF1/CTF to the Nis upstream enhancer region, as well as a clear functional activation of the Nis promoter by both transcription factors. Conclusions/Significance In search for potential downstream mediators of FoxE1 function in thyroid cells, we identified two novel direct FoxE1 target genes. To our knowledge, this is the first evidence regarding the implication of Nis and Duox2 in executing the transcriptional program triggered by FoxE1. Furthermore, this study points out the important role of FoxE1 in the regulation of a large number of genes in thyroid cells. PMID:23675434

  5. 26 CFR 1.642(e)-1 - Depreciation and depletion.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Depreciation and depletion. 1.642(e)-1 Section 1... (CONTINUED) INCOME TAXES Estates, Trusts, and Beneficiaries § 1.642(e)-1 Depreciation and depletion. An estate or trust is allowed the deductions for depreciation and depletion, but only to the extent...

  6. 26 CFR 31.3302(e)-1 - Successor employer.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Successor employer. 31.3302(e)-1 Section 31... Federal Unemployment Tax Act (Chapter 23, Internal Revenue Code of 1954) § 31.3302(e)-1 Successor employer... another person (referred to in this section as a predecessor) who is not an employer (see §...

  7. CYP2E1 Metabolism of Styrene Involves Allostery

    PubMed Central

    Hartman, Jessica H.; Boysen, Gunnar

    2012-01-01

    We are the first to report allosterism during styrene oxidation by recombinant CYP2E1 and human liver microsomes. At low styrene concentrations, oxidation is inefficient because of weak binding to CYP2E1 (Ks = 830 μM). A second styrene molecule then binds CYP2E1 with higher affinity (Kss = 110 μM) and significantly improves oxidation to achieve a kcat of 6.3 nmol · min−1 · nmol CYP2E1−1. The transition between these metabolic cycles coincides with reported styrene concentrations in blood from exposed workers; thus, this CYP2E1 mechanism may be relevant in vivo. Scaled modeling of the in vitro-positive allosteric mechanism for styrene metabolism to its in vivo clearance led to significant deviations from the traditional model based on Michaelis-Menten kinetics. Low styrene levels were notably much less toxic than generally assumed. We interrogated the allosteric mechanism using the CYP2E1-specific inhibitor and drug 4-methylpyrazole, which we have shown binds two CYP2E1 sites. From the current studies, styrene was a positive allosteric effector on 4-methylpyrazole binding, based on a 10-fold increase in 4-methylpyrazole binding affinity from Ki 0.51 to Ksi 0.043 μM. The inhibitor was a negative allosteric effector on styrene oxidation, because kcat decreased 6-fold to 0.98 nmol · min−1 · nmol CYP2E1−1. Consequently, mixtures of styrene and other molecules can induce allosteric effects on binding and metabolism by CYP2E1 and thus mitigate the efficiency of their metabolism and corresponding effects on human health. Taken together, our elucidation of mechanisms for these allosteric reactions provides a powerful tool for further investigating the complexities of CYP2E1 metabolism of drugs and pollutants. PMID:22807108

  8. Application of Strength Diagnosis.

    ERIC Educational Resources Information Center

    Newton, Robert U.; Dugan, Eric

    2002-01-01

    Discusses the various strength qualities (maximum strength, high- and low-load speed strength, reactive strength, rate of force development, and skill performance), noting why a training program design based on strength diagnosis can lead to greater efficacy and better performance gains for the athlete. Examples of tests used to assess strength…

  9. Oncolytic Replication of E1b-Deleted Adenoviruses

    PubMed Central

    Cheng, Pei-Hsin; Wechman, Stephen L.; McMasters, Kelly M.; Zhou, Heshan Sam

    2015-01-01

    Various viruses have been studied and developed for oncolytic virotherapies. In virotherapy, a relatively small amount of viruses used in an intratumoral injection preferentially replicate in and lyse cancer cells, leading to the release of amplified viral particles that spread the infection to the surrounding tumor cells and reduce the tumor mass. Adenoviruses (Ads) are most commonly used for oncolytic virotherapy due to their infection efficacy, high titer production, safety, easy genetic modification, and well-studied replication characteristics. Ads with deletion of E1b55K preferentially replicate in and destroy cancer cells and have been used in multiple clinical trials. H101, one of the E1b55K-deleted Ads, has been used for the treatment of late-stage cancers as the first approved virotherapy agent. However, the mechanism of selective replication of E1b-deleted Ads in cancer cells is still not well characterized. This review will focus on three potential molecular mechanisms of oncolytic replication of E1b55K-deleted Ads. These mechanisms are based upon the functions of the viral E1B55K protein that are associated with p53 inhibition, late viral mRNA export, and cell cycle disruption. PMID:26561828

  10. Acetaldehyde and parkinsonism: role of CYP450 2E1

    PubMed Central

    Vaglini, Francesca; Viaggi, Cristina; Piro, Valentina; Pardini, Carla; Gerace, Claudio; Scarselli, Marco; Corsini, Giovanni Umberto

    2013-01-01

    The present review update the relationship between acetaldehyde (ACE) and parkinsonism with a specific focus on the role of P450 system and CYP 2E1 isozyme particularly. We have indicated that ACE is able to enhance the parkinsonism induced in mice by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, a neurotoxin able to damage the nigrostriatal dopaminergic pathway. Similarly diethyldithiocarbamate, the main metabolite of disulfiram, a drug widely used to control alcoholism, diallylsulfide (DAS) and phenylisothiocyanate also markedly enhance the toxin-related parkinsonism. All these compounds are substrate/inhibitors of CYP450 2E1 isozyme. The presence of CYP 2E1 has been detected in the dopamine (DA) neurons of rodent Substantia Nigra (SN), but a precise function of the enzyme has not been elucidated yet. By treating CYP 2E1 knockout (KO) mice with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, the SN induced lesion was significantly reduced when compared with the lesion observed in wild-type animals. Several in vivo and in vitro studies led to the conclusion that CYP 2E1 may enhance the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxicity in mice by increasing free radical production inside the dopaminergic neurons. ACE is a good substrate for CYP 2E1 enzyme as the other substrate-inhibitors and by this way may facilitate the susceptibility of dopaminergic neurons to toxic events. The literature suggests that ethanol and/or disulfiram may be responsible for toxic parkinsonism in human and it indicates that basal ganglia are the major targets of disulfiram toxicity. A very recent study reports that there are a decreased methylation of the CYP 2E1 gene and increased expression of CYP 2E1 mRNA in Parkinson's disease (PD) patient brains. This study suggests that epigenetic variants of this cytochrome contribute to the susceptibility, thus confirming multiples lines of evidence which indicate a link between environmental toxins and PD. PMID:23801948

  11. Multi-Laboratory Validation of Estrone (E1) ELISA Methods

    EPA Science Inventory

    This project is a round-robin evaluation of commercially available Enzyme-Linked Immunosorbent Assay (ELISA) technology to quantitatively or qualitatively measure the hormone estrone (E1) in combined animal feeding operation (CAFO) receiving streams. ELISA is meant to be a simpl...

  12. 24. SPILLWAY CHANNEL WALLS REINFORCEMENT DETAILS; MONOLITHS E1 TO ...

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

    24. SPILLWAY CHANNEL WALLS - REINFORCEMENT DETAILS; MONOLITHS E-1 TO F-4 INCL. & NO. 34. Sheet S-11, June, 1939. File no. SA 342/24(?). - Prado Dam, Spillway, Santa Ana River near junction of State Highways 71 & 91, Corona, Riverside County, CA

  13. Quantitation of ColE1-encoded replication elements.

    PubMed Central

    Brenner, M; Tomizawa, J

    1991-01-01

    Replication of the Escherichia coli plasmid ColE1 initiates from an RNA primer. This primer is formed by a ColE1 RNA II molecule that remains hybridized to its DNA template in the origin region after transcription. Continued hybridization is inhibited by prior binding to RNA II of another ColE1 transcript, RNA I; and this interaction is regulated by the plasmid-encoded Rom protein. To understand the quantitative aspects of regulation of ColE1 synthesis, we have measured the levels of RNA I, RNA II, and Rom protein in vivo, as well as the half-lives of the RNAs. The intracellular concentrations of RNA I, RNA II, and Rom protein were found to be about 1 microM, 7 nM, and 1 microM, respectively; and the RNAs had half-lives of about 2 min. A simple model derived from these results indicates that the plasmid copy number is little affected by the rate of RNA II synthesis but is strongly dependent on that of RNA I. Images PMID:1703297

  14. EXPRESSION OF ENDOGLUCANASE E1 IN TRANSGENIC DUCKWEED LEMNA MINOR

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transgenic duckweed (Lemna minor) that expresses Acidothermus cellulolyticus E1 endoglucanase was generated using Agrobacterium-mediated transformation. Out of 15 independent transgenic lines, 1 line with the highest CMC-degrading activity was selected for further studies. The 2-week-old transgeni...

  15. Induction of cyp2e-1 protein in mouse colon.

    PubMed

    Rosenberg, D W; Mankowski, D C

    1994-01-01

    The inducibility of a cytochrome P450 isoform, CYP2E1 (cyp2e-1), was compared in colonic epithelium of selected inbred mice. Mice were chosen for study on the basis of reported susceptibility to 1,2-dimethylhydrazine (DMH)-induced colorectal tumor formation. DBA/2J (resistant), C57BL/6J (intermediate) and SWR/J (susceptible) mice were exposed to acetone (1% v/v) in drinking water for 10 days. SWR/J mice sustained the largest increase in colonic cyp2e-1, although protein levels, assessed by Western analysis, were markedly increased in mucosal tissue obtained from C57BL/6J mice as well. Further evidence for colonic cyp2e-1 induction is supported by elevated (3.5-fold) chlorzoxazone 6-hydroxylase activity in response to acetone. To more fully characterize these changes in colon, the tumor-sensitive SWR/J mice were chosen for further evaluation. Mice were treated with a panel of agents established to induce this protein in liver, including isoniazid (0.1% v/v) and ethanol (10% v/v) in drinking water and pyrazole (300 mg/kg), given intraperitoneally. With the exception of ethanol, each compound produced a marked (1.5- to 3-fold) elevation of cyp2e-1 in colon and liver. Overall balance between phase I and II metabolism may be a critical factor in determining tumor susceptibility. Therefore, glutathione S-transferase (GST) activity was also examined. In liver, basal GST levels varied less than 2-fold between strains, while in colon, levels were 5-10% of corresponding hepatic levels. Although acetone treatment did not significantly alter hepatic GST, a 30-60% decline in activity was observed in colons of SWR/J and C57BL/6J mice. Further examination of colonic GST revealed compound-specific effects. Ethanol exposure markedly (60%) lowered GST levels in colon, whereas pyrazole produced a 2-fold increase. None of these agents significantly altered hepatic GST activity. These studies demonstrate the ability of mouse colon to undergo an increase in immunoreactive cyp2e-1 in response to a panel of xenobiotics known to elevate this protein in liver. Further characterization of cyp2e-1 and GSTs in inbred mice may provide important information on the role of colonocytes in direct activation of ingested procarcinogens to DNA-reactive metabolites. PMID:8293551

  16. Cyclin E1 Is Amplified and Overexpressed in Osteosarcoma

    PubMed Central

    Lockwood, William W.; Stack, Deirdre; Morris, Thomas; Grehan, David; O'Keane, Conor; Stewart, Greg L.; Cumiskey, Joanna; Lam, Wan L.; Squire, Jeremy A.; Thomas, David M.; O'Sullivan, Maureen J.

    2011-01-01

    Osteosarcoma is a genetically complex malignancy, predominantly afflicting the adolescent population and associated still with relatively poor long-term outcomes. Although there has been some improvement in the understanding of osteosarcoma biology, this has not yet translated particularly well into therapeutic advances. By using a whole-genome tiling path array for comparative genomic hybridization analysis, we sought to evaluate DNA copy number changes in 22 osteosarcoma tumor samples. Regions of most frequent gains or losses generated by Genomic Identification of Significant Targets in Cancer analysis were evaluated for genes of interest. Correlation of the copy number data with preexisting expression data for these genes yielded not only targets known to be important in osteosarcoma but also novel targets, notably cyclin E1. Fluorescence in situ hybridization and immunohistochemical analysis confirmed the findings. Overexpression of cyclin E1 has potential prognostic and therapeutic implications that are discussed herein. PMID:21458381

  17. Anharmonic mixing between two phonons in red mercury iodide

    NASA Astrophysics Data System (ADS)

    Anedda, A.; Bongiovanni, G.

    1988-12-01

    With use of Raman spectroscopy we have investigated the anharmonic mixing between two optical phonons of Eg symmetry in red mercury iodide. By taking into account the presence of several decay channels contributing to phonon lifetime, the experimental line-shape anomalies due to mode coupling have been reproduced with good accuracy. The temperature behavior of the anharmonic interaction is in agreement with theory.

  18. Flexibility and Muscular Strength.

    ERIC Educational Resources Information Center

    Liemohn, Wendell

    1988-01-01

    This definition of flexibility and muscular strength also explores their roles in overall physical fitness and focuses on how increased flexibility and muscular strength can help decrease or eliminate lower back pain. (CB)

  19. Treatment of vasospastic disease with prostaglandin E1.

    PubMed Central

    Clifford, P C; Martin, M F; Sheddon, E J; Kirby, J D; Baird, R N; Dieppe, P A

    1980-01-01

    Prostaglandin E1, a vasodilator and potent inhibitor of platelet aggregation, was administered to 26 patients with severe vasospastic disease of the hands. Patients tolerated infusions well and reported appreciable symptomatic improvement. Five of eight ischaemic ulcers healed in six weeks. Non-invasive studies of blood flow were used to observe haemodynamic changes during and after infusions. The Doppler-derived radial artery pulsatility index fell from 8.8 +/- 0.6 to 4.6 +/0 0.5 (mean +/- SEM), indicating hand vasodilatation. This fall was maintained 24 hours after infusion (5.9 +/- 0.9), but the index had returned to normal values at two weeks. The amplitude of finger pulse volume recordings increased (5.6 +/- 0.7 mm to 23.8 +/- 3.4 mm) and was raised two and six weeks after infusion (13.5 +/- 2.1 mm). Hand temperature measured by infrared radiometry also increased (27.4 +/- 0.7 degrees C to 31.2 +/- 1.2 degrees C). Intensity of digital vasospasm induced by cold water challenge was not objectively affected by prostaglandin E1 despite an increased finger temperature after infusion. Nevertheless, patients reported less frequent and severe attacks. Prostaglandin E1 given by central venous infusion is a safe new vasoactive agent that can produce appreciable symptomatic improvement by increasing digital perfusion, which may last for several weeks after treatment. Further study will define its mode of action and its place in the management of peripheral vascular disease. PMID:7427564

  20. Octupole deformation in sup 221 Fr; E1 transition rates

    SciTech Connect

    Liang, C.F.; Peghaire, A. ); Sheline, R.K. )

    1990-07-10

    Experimental data following the alpha decay of{sup 225}Ac are interpreted in terms of a spectroscopy in {sup 221}Fr consistent with octupole deformation. However, the measured E1 transition probabilities suggest that the low lying bands in {sup 221}Fr are considerably more mixed than in nuclei with slightly higher mass number. It is suggested that this mixing of states in {sup 221}Fr is indicative of the partial collapse of Nilsson-like orbitals into more degenerate shell model orbitals.

  1. RP-1 delivered to E-1 Test Stand

    NASA Technical Reports Server (NTRS)

    2010-01-01

    NASA John C. Stennis Space Center employee Dustan Ladner (left) assists tanker driver David Velasco in transferring RP-1 fuel to a 20,000-gallon underground tank at the E-1 Test Stand during a March 30 delivery. The rocket propellant will be used for testing Aerojet AJ26 rocket engines beginning this summer. Stennis is testing the engines for Orbital Sciences Corporation, which has partnered with NASA to provide eight supply missions to the International Space Station through 2015. The partnership is part of NASA's Commercial Orbital Transportation Services initiative to work closer with companies to provide commercial space transport once the space shuttle is retired later this year.

  2. Haemorrheological effects of prostaglandin E1 infusion in Raynaud's syndrome.

    PubMed Central

    Lucas, G S; Simms, M H; Caldwell, N M; Alexander, S J; Stuart, J

    1984-01-01

    Eighteen patients with severe Raynaud's syndrome had impaired deformability of erythrocytes, as measured by filtration through 5 micron diameter pores, compared with 19 healthy controls. The patients were given prostaglandin E1 (PGE1) or placebo by intravenous infusion for 72 h to assess the haemorrheological action of PGE1. Contrary to a previous report, PGE1 did not improve erythrocyte filterability. Infusion of PGE1 did, however, evoke an acute phase response with hyperproteinaemia and a leucocytosis and is a potentially important mediator of this stress response in patients with vascular disease. PMID:6540787

  3. A large scale virtual screen of DprE1.

    PubMed

    Wilsey, Claire; Gurka, Jessica; Toth, David; Franco, Jimmy

    2013-12-01

    Tuberculosis continues to plague the world with the World Health Organization estimating that about one third of the world's population is infected. Due to the emergence of MDR and XDR strains of TB, the need for novel therapeutics has become increasing urgent. Herein we report the results of a virtual screen of 4.1 million compounds against a promising drug target, DrpE1. The virtual compounds were obtained from the Zinc docking site and screened using the molecular docking program, AutoDock Vina. The computational hits have led to the identification of several promising lead compounds. PMID:24055764

  4. Phenomenology of heavy quarkonium radiative E1 transitions

    NASA Astrophysics Data System (ADS)

    Martinez, Hector E.

    2016-01-01

    We present preliminary results of the evaluation of the next-to-leading-order (NLO) relativistic corrections to the heavy quarkonium electric dipole transition (E1) rate. In our evaluation we use the quark-antiquark potential up to 1/m2 corrections that includes the effective string theory expression for the long range, a review on the method to construct this potential is given. Our results compare favorable with the experiments and may provide predictions for the rates for which no experimental data is yet available.

  5. Strength Training for Girls.

    ERIC Educational Resources Information Center

    Connaughton, Daniel; Connaughton, Angela; Poor, Linda

    2001-01-01

    Strength training can be fun, safe, and appropriate for young girls and women and is an important component of any fitness program when combined with appropriate cardiovascular and flexibility activities. Concerns and misconceptions regarding girls' strength training are discussed, presenting general principles of strength training for children…

  6. Crossover Analysis of CHANG'E-1 Laser Altimeter Data

    NASA Astrophysics Data System (ADS)

    Hu, W.; Yue, Z.; Di, K.

    2011-08-01

    This paper presents a preliminary result of crossover analysis and adjustment of Chang'E-1(CE-1) Laser Altimeter (LAM) data of the Moon for global and regional mapping applications. During the operation of Chang'E-1 from November 28, 2007 to December 4, 2008, the laser altimeter acquired 1400 orbital profiles with about 9.12 million altimetric points. In our experiment, we derived more than 1.38 million crossovers from 1395 ground tracks covering the entire lunar surface after eliminating outliers of orbits and altimetric points. A method of least-squares crossover adjustment with a series of basis functions of time (trigonometric functions and polynomials) is developed to reconcile the LAM data by minimizing the crossover residuals globally. The normal equations are very large but sparse; therefore they are stored and solved using sparse matrix technique. In a test area (0°N~60°N, 50°W~0°W), the crossover residuals are reduced from 62.1m to 32.8m, and the quality of the DEM generated from the adjusted LAM data is improved accordingly. We will optimize the method for the global adjustment to generate a high precision consistent global DEM, which can be used as absolute control for lunar mapping with orbital images.

  7. A Portable E1-M1 Optical Clock

    NASA Astrophysics Data System (ADS)

    Alden, Emily; Leanhardt, Aaron

    2013-05-01

    The frontiers of precision time measurement are improving rapidly. The pursuit of the lowest clock stability has produced a measurement of time dilation from to an elevation change of less than a meter and creates the potential for tabletop tests of fundamental constants. We present a new setup for an optical frequency standard that accesses the E 1 forbidden 1S0 -->3P0 clock transition by a two-photon allowed E 1 - M 1 clock transition along 1S0 -->3P1 -->3P0 . This pathway has the potential to permit detectable clock transitions for atoms at room temperature. Further, the transition can be driven using Doppler-free spectroscopy techniques which immediately remove the dominant broadening mechanism for single-photon clocks at room temperature. Operating with hot atoms removes the extensive state preparation required by other optical frequency standards and thus makes the clock more portable that ion or lattice systems. We present the basic components of this novel technique and progress toward a hot neutral Hg clock. This work has been supported by DARPA and AFOSR.

  8. Strength Modeling Report

    NASA Technical Reports Server (NTRS)

    Badler, N. I.; Lee, P.; Wong, S.

    1985-01-01

    Strength modeling is a complex and multi-dimensional issue. There are numerous parameters to the problem of characterizing human strength, most notably: (1) position and orientation of body joints; (2) isometric versus dynamic strength; (3) effector force versus joint torque; (4) instantaneous versus steady force; (5) active force versus reactive force; (6) presence or absence of gravity; (7) body somatotype and composition; (8) body (segment) masses; (9) muscle group envolvement; (10) muscle size; (11) fatigue; and (12) practice (training) or familiarity. In surveying the available literature on strength measurement and modeling an attempt was made to examine as many of these parameters as possible. The conclusions reached at this point toward the feasibility of implementing computationally reasonable human strength models. The assessment of accuracy of any model against a specific individual, however, will probably not be possible on any realistic scale. Taken statistically, strength modeling may be an effective tool for general questions of task feasibility and strength requirements.

  9. Metabolism of prostaglandins A1 and E1 in man.

    PubMed Central

    Golub, M; Zia, P; Matsuno, M; Horton, R

    1975-01-01

    To investigate the in vivo whole blood metabolic clearance rates and sites of metabolism of prostaglandins A1 and E1 in man, constant infusions of the tritiated compounds were administered to normal subjects and to patients undergoing cardiac catheterization. The whole blood metabolic clearance rate of [3H]prostaglandin A1 in eight men was 5,003 +/- 864 liters/day (SD) or 2,546 +/- 513 liters/day per m2 (SD). Nonradioactive prostaglandin A1 was similarly infused in two subjects, and the metabolic clearance rates were determined, utilizing a specific radioimmunoassay. The clearance rates with this method correlated closely with those determined by the isotope infusions. Extraction studies of prostaglandin A1 showed that pulmonary, splanchnic, renal, and extremity perfusions resulted in 8.1 +/- 4.1, 56.1 +/- 10.1, 50.3 +/- 3.4, and 34.4 +/- 5.9% (SEM) removal, respectively. With [3H]=prostaglandin E1, the whole blood metabolic clearance rate was determined from the pulmonary artery concentration in three patients and averaged 4,832 +/- 1,518 liters/day (SD) or 2,686 +/- 654 liters/day per m2 (SD). Pulmonary extraction was 67.8 +/- 6.8% (SEM) and extremity removal averaged 6.6 +/- 4.9% (SEM). These results indicate that A prostaglandins are metabolized by several organs, such as the liver and kidney, and possibly by intravascular pathways as well. In man, the E prostaglandins are primarily metabolized by the lung, but extraction is not complete and approximately one-third may escape lung metabolism. Thus, these findings suggest that both E and A prostaglandins in the venous circulation may reach the systemic circulation in man. PMID:1202078

  10. Alumina fiber strength improvement

    NASA Technical Reports Server (NTRS)

    Pepper, R. T.; Nelson, D. C.

    1982-01-01

    The effective fiber strength of alumina fibers in an aluminum composite was increased to 173,000 psi. A high temperature heat treatment, combined with a glassy carbon surface coating, was used to prevent degradation and improve fiber tensile strength. Attempts to achieve chemical strengthening of the alumina fiber by chromium oxide and boron oxide coatings proved unsuccessful. A major problem encountered on the program was the low and inconsistent strength of the Dupont Fiber FP used for the investigation.

  11. Effects of Colicins E1 and K on Cellular Metabolism

    PubMed Central

    Fields, Kay L.; Luria, S. E.

    1969-01-01

    Colicins E1 and K inhibited a whole series of energy-dependent reactions in Escherichia coli cells, including motility, biosynthesis of nucleic acids, proteins and polysaccharides, and the conversion of ornithine to citrulline. Respiration was only partially affected, and substrates such as glucose continued to be catabolized through the normal pathways, albeit with reduced CO2 production. The soluble products of aerobic glucose catabolism by colicin-treated cells were analyzed. Pyruvate replaced acetate as the major excreted product, and the following intermediates of glycolysis were excreted in significant amounts: glucose-6-phosphate, fructose-1,6-diphosphate, dihydroxyacetone phosphate, and 3-phosphoglycerate. Anaerobically growing cells manifested a somewhat enhanced tolerance to the colicins. This protection by anaerobiosis appeared to depend on the exclusion of oxygen more than on the extent of fermentative catabolism versus catabolism of the respiratory type. These results are interpreted in terms of possible functions of colicin in lowering the adenosine triphosphate (ATP) content of the cells and in terms of the role of lowered ATP levels in inhibiting many of the energy-requiring reactions. PMID:4884824

  12. Enhanced reliability features of the RL10E-1 engine

    NASA Astrophysics Data System (ADS)

    Van Lerberghe, W. M.; Emdee, J. L.; Foust, R. R.

    The RLlOE-1 rocket engine has been partially developed for the United States Air Force during the Atlas Reliability Enhancement Program. This engine is a 22,300 Ibf thrust liquid hydrogen, liquid oxygen, derivative of the RL10A-4-1, and incorporates an improved ignition system and new electromechanically actuated (EMA) valves controlled by an engine mounted Digital Electronic Rocket Engine Controller. The RLlOE-1 electronic control system operates the engine with only six valves (compared to the twelve on the RLIOA-4-1), provides boost phase engine cooldown capability (to avoid cryogenic temperatures at liftoff), and allows implementation of an improved torch ignition system with redundant, modem electronics. Other potential reliability improvements include active engine health monitoring, derated thrust operation, "soft" transients, and improved minimum residual shutdown capability. Although development of the RL10E-1 was not completed, the engine development tests were highly successful in demonstrating engine operation with the full authority, digital electronic control system and fewer valves. Many of the enhanced reliability features have been demonstrated and are directly applicable to other versions of the RL10 engines. Potential "hybrid" engine configurations offer a lower weight and less costly engine, while retaining much of the reliability improvements and other benefits of the RLIOE-1.

  13. Human adenovirus 2 E1B-19K and E1B-53K tumor antigens: antipeptide antibodies targeted to the NH2 and COOH termini.

    PubMed Central

    Green, M; Brackmann, K H; Lucher, L A; Symington, J S; Kramer, T A

    1983-01-01

    The human adenovirus 2 (Ad2) transforming region is located in the left 11.1% of the viral genome and encodes two early transcription units, E1A and E1B. Based on the amino acid sequence deduced from the Ad2 E1B DNA sequence (Gingeras et al., J. Biol. Chem. 257:13475-13491, 1982), we have prepared antibodies against synthetic peptides, 8 to 16 amino acids in length, encoded at the NH2 and COOH termini of the major E1B-19K and E1B-53K tumor antigens. The antipeptide antibodies immunoprecipitated the targeted E1B-19K or E1B-53K tumor antigens from extracts of Ad2-infected cells. The specificity of the peptide competition studies. Antipeptide antibodies directed to the NH2 and COOH termini immunoprecipitated the E1B-19K and E1B-53K tumor antigens from two Ad2-transformed rat cell lines, F17 and F4, providing evidence that identical tumor antigens are synthesized in Ad2-infected and Ad2-transformed cells. These results show that the E1B-19K and E1B-53K T antigens are not processed proteolytically at either the NH2 or COOH terminus. Our data provide strong evidence at the protein level that the E1B-19K and E1B-53K tumor antigens partially overlap in DNA sequence, with the E1B-19K initiating translation at the first ATG at nucleotide 1711 in translation reading frame 1 and the E1B-53K tumor antigen initiating translation at the second ATG at nucleotide 2016 in reading frame 3. This confirms the results of others on the N-terminal amino acid sequence of E1B-19K and theoretical deductions based on the DNA sequence. Our findings prove that the large E1B-53K T antigen initiates translation at the second ATG at nucleotide 2016 and not at equally plausible initiation codons located farther downstream at nucleotides 2202 and 2235. Thus, the E1B-53K T antigen is another example of a protein which initiates translation at an internal ATG rather than at the 5'-proximal ATG. Images PMID:6632083

  14. High strength, superplastic superalloy

    NASA Technical Reports Server (NTRS)

    Ashbrook, R. L.; Freche, J. C.; Waters, W. J.

    1969-01-01

    High strength superplastic superalloys are produced by extruding a pre-alloyed powder. The cast nickel base superalloy was remelted and converted to pre-alloyed powder by inert gas atomization. The superalloy shows high tensile strength and superplasticity and finds use in hot working and casting.

  15. Recognizing Neglected Strengths

    ERIC Educational Resources Information Center

    Sternberg, Robert J.

    2006-01-01

    To identify diverse student strengths and to learn how teachers can build instruction on those strengths, the author and his colleagues have conducted multiple studies among students in Alaska, the mainland United States, Kenya, and other countries. In a series of studies in Alaska and Kenya, the researchers measured the adaptive cultural…

  16. CNO cycle: ”Soft E1” mode of the 17Ne excitation in the 17Ne+γ → 15O+2p reaction

    NASA Astrophysics Data System (ADS)

    Parfenova, Yu L.; Grigorenko, L. V.; Egorova, I. A.; Shulgina, N. B.; Zhukov, M. V.

    2016-01-01

    The 15O(2p, γ)17Ne reaction is studied using the time-reversed reaction of the17Ne E1 Coulomb dissociation on lead target in the context of nuclear astrophysics. Looking for the relation between the data on the Coulomb excitation and the astrophysical 2p-capture rate, one faces problem to extract the Coulomb E1 strength function from the measured Coulex cross section. We use a number of phenomenological approaches to estimate influence of such processes as Coulomb-nuclear interference, populations of states with different Jπ, etc. We calculate the 17Ne+2p astrophysical capture rate and compare the results with different calculations.

  17. The dual effect of adenovirus type 5 E1A 13S protein on NF-kappaB activation is antagonized by E1B 19K.

    PubMed Central

    Schmitz, M L; Indorf, A; Limbourg, F P; Städtler, H; Traenckner, E B; Baeuerle, P A

    1996-01-01

    The genomes of human adenoviruses encode several regulatory proteins, including the two differentially spliced gene products E1A and E1B. Here, we show that the 13S but not the 12S splice variant of E1A of adenovirus type 5 can activate the human transcription factor NF-kappaB in a bimodal fashion. One mode is the activation of NF-kappaB containing the p65 subunit from the cytoplasmic NF-kappaB-IkappaB complex. This activation required reactive oxygen intermediates and the phosphorylation of IkappaBalpha at serines 32 and 36, followed by IkappaBalpha degradation and the nuclear uptake of NF-kappaB. In addition, 13S E1A stimulated the transcriptional activity of the C-terminal 80 amino acids of p65 at a core promoter with either a TATA box or an initiator (INR) element. The C-terminal 80 amino acids of p65 were found to associate with E1A in vitro. The activation of NF-kappaB-dependent reporter gene transcription by E1A was potently suppressed upon coexpression of the E1B 19-kDa protein (19K). E1B 19K prevented both the activation of NF-kappaB and the E1A-mediated transcriptional enhancement of p65. These inhibitory effects were not found for the 55-kDa splice variant of the E1B protein. We suggest that the inductive effect of E1A 13S on the host factor NF-kappaB, whose activation is important for the transcription of various adenovirus genes, must be counteracted by the suppressive effect of E1B 19K so that the adenovirus-infected cell can escape the immune-stimulatory and apoptotic effects of NF-kappaB. PMID:8754803

  18. The adenovirus E1A proteins induce apoptosis, which is inhibited by the E1B 19-kDa and Bcl-2 proteins.

    PubMed Central

    Rao, L; Debbas, M; Sabbatini, P; Hockenbery, D; Korsmeyer, S; White, E

    1992-01-01

    Cooperation between the adenovirus E1A and E1B oncogenes is required for transformation of primary quiescent rodent cells. Although expression of E1A alone will stimulate cell proliferation sufficient to initiate transformed focus formation, proliferation fails to be sustained and foci degenerate. Coexpression of either the 19-kDa or 55-kDa E1B oncoproteins with E1A permits high-frequency transformation by overcoming this cytotoxic response. Without E1B 19-kDa protein expression, however, transformants remain susceptible to induction of cell death. Rapid loss of viability is coincident with nucleolytic cleavage of DNA in intranucleosomal regions and chromatin condensation, hallmarks of programmed cell death (apoptosis). Furthermore, overexpression of a known suppressor of apoptosis, the Bcl-2 protooncogene, can rescue E1A-induced focus degeneration. Thus E1A-dependent stimulation of cell proliferation is accompanied by apoptosis and thereby insufficient to singly induce transformation. High-frequency transformation requires a second function encoded by the E1B 19-kDa protein to block apoptosis. Images PMID:1457005

  19. Sequence-independent autoregulation of the adenovirus type 5 E1A transcription unit.

    PubMed Central

    Hearing, P; Shenk, T

    1985-01-01

    The adenovirus E1A gene is known to be autoregulated at the level of transcription. Autoregulation was found to be mediated by products of the E1A 13S mRNA, which induced a fivefold increase in E1A transcription rate. Deletion analysis suggested that the autoregulation did not require any specific sequence in the E1A transcriptional control region. This conclusion was reinforced by the demonstration that a cellular alpha-globin gene substituted for the E1A gene on the adenovirus chromosome was also positively regulated by E1A gene products. Images PMID:2943984

  20. Strength and Balance Exercises

    MedlinePlus

    ... Blood Pressure Tools & Resources Stroke More Strength and Balance Exercises Updated:Jun 23,2015 If you have ... second set. Walking Heel-to-Toe Purpose: Improve balance. Starting Position: Stand close to a wall for ...

  1. Reduction of bone strength

    NASA Technical Reports Server (NTRS)

    Bingham, Cindy

    1990-01-01

    Viewgraphs on reduction of bone strength are presented. WEHI 231 B growth rates, experimental chambers used to apply the electric field to the cell cultures, and a mouse suspended by rotating cuff in electromagnetic field are shown.

  2. Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting

    SciTech Connect

    Dai, Ziyu; Hooker, Brian S.; Anderson, Daniel B.; Thomas, S R.

    2000-06-01

    Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up to 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggests dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.

  3. Hand-Strength Meter

    NASA Technical Reports Server (NTRS)

    Tcheng, Ping; Elliot, Joe

    1987-01-01

    Special grip-strength meter designed for accurate, reproducible measurement of hand rehabilitation. Four strain gauges connected in Wheatstone bridge to measure deflection caused by gripping hand. Compressive force exerted by hand transmitted to measuring beams. Beams therefore deflected or strained, and mechanical strain sensed by strain gauges and converted into electrical signal. After amplification and conditioning, signal displayed on LED as measure of gripping strength of hand.

  4. Apple Strength Issues

    SciTech Connect

    Syn, C

    2009-12-22

    Strength of the apple parts has been noticed to decrease, especially those installed by the new induction heating system since the LEP campaign started. Fig. 1 shows the ultimate tensile strength (UTS), yield strength (YS), and elongation of the installed or installation-simulated apples on various systems. One can clearly see the mean values of UTS and YS of the post-LEP parts decreased by about 8 ksi and 6 ksi respectively from those of the pre-LEP parts. The slight increase in elongation seen in Fig.1 can be understood from the weak inverse relationship between the strength and elongation in metals. Fig.2 shows the weak correlation between the YS and elongation of the parts listed in Fig. 1. Strength data listed in Figure 1 were re-plotted as histograms in Figs. 3 and 4. Figs. 3a and 4a show histograms of all UTS and YS data. Figs. 3b and 4b shows histograms of pre-LEP data and Figs. 3c and 4c of post-LEP data. Data on statistical scatter of tensile strengths have been rarely published by material suppliers. Instead, only the minimum 'guaranteed' strength data are typically presented. An example of strength distribution of aluminum 7075-T6 sheet material, listed in Fig. 5, show that its scatter width of both UTS and YS for a single sheet can be about 6 ksi and for multi-lot scatter can be as large as 11 ksi even though the sheets have been produced through well-controlled manufacturing process. By approximating the histograms shown in Figs. 3 and 4 by a Gaussian or similar type of distribution curves, one can plausibly see the strength reductions in the later or more recent apples. The pre-LEP data in Figs. 3b and 4b show wider scatter than the post-LEP data in Figs. 3c and 4c and seem to follow the binomial distribution of strength indicating that the apples might have been made from two different lots of material, either from two different vendors or from two different melts of perhaps slightly different chemical composition by a single vendor. The post-LEP apples seem to have been from a single batch of material. The pre-LEP apples of the weak strength and the post-LEP apples with even weaker strength could have been made of the same batch of material, and the small strength differential might be due to the difference in the induction heating system. If the pre-LEP apples with the lower strength and the post LEP apples are made from the same batch of material, their combined scatter of strength data would be wider and can be understood as a result of the additional processing steps of stress relief and induction heating as discussed.

  5. 26 CFR 48.4216(e)-1 - Exclusion of local advertising charges from sale price.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... sale price. 48.4216(e)-1 Section 48.4216(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... Provisions Applicable to Manufacturers Taxes § 48.4216(e)-1 Exclusion of local advertising charges from sale price. (a) In general. Section 4216(e) deals with the treatment to be accorded charges made by...

  6. 26 CFR 1.501(e)-1 - Cooperative hospital service organizations.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ....501(e)-1 Section 1.501(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(e)-1 Cooperative hospital service organizations. (a) General rule. Section 501(e) is the exclusive and controlling...

  7. 26 CFR 48.4216(e)-1 - Exclusion of local advertising charges from sale price.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... sale price. 48.4216(e)-1 Section 48.4216(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... Provisions Applicable to Manufacturers Taxes § 48.4216(e)-1 Exclusion of local advertising charges from sale price. (a) In general. Section 4216(e) deals with the treatment to be accorded charges made by...

  8. 26 CFR 1.501(e)-1 - Cooperative hospital service organizations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ....501(e)-1 Section 1.501(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(e)-1 Cooperative hospital service organizations. (a) General rule. Section 501(e) is the exclusive and controlling...

  9. 26 CFR 1.501(e)-1 - Cooperative hospital service organizations.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 7 2014-04-01 2013-04-01 true Cooperative hospital service organizations. 1.501(e)-1 Section 1.501(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Exempt Organizations § 1.501(e)-1 Cooperative hospital service organizations. (a) General...

  10. 26 CFR 48.4216(e)-1 - Exclusion of local advertising charges from sale price.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... sale price. 48.4216(e)-1 Section 48.4216(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... Provisions Applicable to Manufacturers Taxes § 48.4216(e)-1 Exclusion of local advertising charges from sale price. (a) In general. Section 4216(e) deals with the treatment to be accorded charges made by...

  11. 26 CFR 48.4216(e)-1 - Exclusion of local advertising charges from sale price.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... price. 48.4216(e)-1 Section 48.4216(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... Applicable to Manufacturers Taxes § 48.4216(e)-1 Exclusion of local advertising charges from sale price. (a) In general. Section 4216(e) deals with the treatment to be accorded charges made by a...

  12. Identification of a nuclear export signal sequence for bovine papillomavirus E1 protein

    SciTech Connect

    Rosas-Acosta, German; Wilson, Van G.

    2008-03-30

    Recent studies have demonstrated nuclear export by papillomavirus E1 proteins, but the requisite export sequence(s) for bovine papillomavirus (BPV) E1 were not defined. In this report we identify three functional nuclear export sequences (NES) present in BPV E1, with NES2 being the strongest in reporter assays. Nuclear localization of BPV1 E1 was modulated by over- or under-expression of CRM1, the major cellular exportin, and export was strongly reduced by the CRM1 inhibitor, Leptomycin B, indicating that E1 export occurs primarily through a CRM1-dependent process. Consistent with the in vivo functional results, E1 bound CRM1 in an in vitro pull-down assay. In addition, sumoylated E1 bound CRM1 more effectively than unmodified E1, suggesting that E1 export may be regulated by SUMO modification. Lastly, an E1 NES2 mutant accumulated in the nucleus to a greater extent than wild-type E1, yet was defective for viral origin replication in vivo. However, NES2 exhibited no intrinsic replication defect in an in vitro replication assay, implying that nucleocytoplasmic shuttling may be required to maintain E1 in a replication competent state.

  13. 26 CFR 1.1397E-1 - Qualified zone academy bonds.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Qualified zone academy bonds. 1.1397E-1 Section 1.1397E-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Empowerment Zone Employment Credit § 1.1397E-1 Qualified zone...

  14. 26 CFR 1.401(e)-1 - Definitions relating to plans covering self-employed individuals.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...-employed individuals. 1.401(e)-1 Section 1.401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.401(e)-1 Definitions relating to plans covering self-employed...

  15. 26 CFR 1.401(e)-1 - Definitions relating to plans covering self-employed individuals.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...-employed individuals. 1.401(e)-1 Section 1.401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.401(e)-1 Definitions relating to plans covering self-employed...

  16. 26 CFR 1.401(e)-1 - Definitions relating to plans covering self-employed individuals.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...-employed individuals. 1.401(e)-1 Section 1.401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Pension, Profit-Sharing, Stock Bonus Plans, Etc. § 1.401(e)-1 Definitions relating to plans covering self-employed...

  17. 26 CFR 1.149(e)-1 - Information reporting requirements for tax-exempt bonds.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...-exempt bonds. 1.149(e)-1 Section 1.149(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE... and Local Bonds § 1.149(e)-1 Information reporting requirements for tax-exempt bonds. (a) General rule... that issuer (e.g., previous affidavits and certifications for other private activity bonds issued...

  18. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Regulations 118 (26 CFR part 39, 1939 ed.) for the necessary amount. On January 1, 1965, B, a calendar year... when prior law applied. 1.691(e)-1 Section 1.691(e)-1 Internal Revenue INTERNAL REVENUE SERVICE....691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In...

  19. 26 CFR 1.924(e)-1 - Activities relating to the disposition of export property.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... export property. 1.924(e)-1 Section 1.924(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... United States § 1.924(e)-1 Activities relating to the disposition of export property. (a) Advertising and sales promotion. For purposes of section 924(e), advertising and sales promotion are defined as...

  20. 26 CFR 1.669(e)-1A - Pro rata portion of taxes deemed distributed.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Pro rata portion of taxes deemed distributed. 1.669(e)-1A Section 1.669(e)-1A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... Taxable Years Beginning Before January 1, 1969 § 1.669(e)-1A Pro rata portion of taxes deemed...

  1. 26 CFR 1.1402(e)-1A - Application of regulations under section 1402(e).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 12 2010-04-01 2010-04-01 false Application of regulations under section 1402(e). 1.1402(e)-1A Section 1.1402(e)-1A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Tax on Self-Employment Income §...

  2. 26 CFR 1.1402(e)-1A - Application of regulations under section 1402(e).

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 12 2011-04-01 2011-04-01 false Application of regulations under section 1402(e). 1.1402(e)-1A Section 1.1402(e)-1A Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Tax on Self-Employment Income §...

  3. Role of E1B55K in E4orf6/E1B55K E3 Ligase Complexes Formed by Different Human Adenovirus Serotypes

    PubMed Central

    Cheng, Chi Ying; Gilson, Timra; Wimmer, Peter; Schreiner, Sabrina; Ketner, Gary; Dobner, Thomas; Branton, Philip E.

    2013-01-01

    The E4orf6 protein of serotypes representing all human adenovirus species forms Cullin-based E3 ubiquitin ligase complexes that facilitate virus infection by inducing degradation of cellular proteins that impede efficient viral replication. This complex also includes the viral E1B55K product believed to bind and introduce substrates for ubiquitination. Heterogeneity in the composition of these ligases exists, as some serotypes form Cul5-based complexes whereas others utilize Cul2. Significant variations in substrate specificities also exist among serotypes, as some degrade certain substrates very efficiently whereas others induce more modest or little degradation. As E1B55K is believed to function as the substrate acquisition component of the ligase, we undertook studies to compare the ability of representative E1B55K proteins to bind substrates with the efficacy of degradation by their respective E4orf6-based ligases. Interestingly, although efficient degradation in some cases corresponded to the ability of E1B55K to bind to or relocalize substrates, there were several examples of substrates that bound efficiently to E1B55K but were not degraded and others in which substrates were degraded even though binding to E1B55K was low or undetectable. These results suggest that transient interactions with E1B55K may be sufficient for efficient substrate degradation and that binding alone is not sufficient, implying that the orientation of the substrate in the ligase complex is probably crucial. Nevertheless, we found that the substrate specificity of certain E4orf6-based ligases could be altered through the formation of hybrid complexes containing E1B55K from another serotype, thus confirming identification of E1B55K as the substrate acquisition component of the complex. PMID:23536656

  4. Spin resonance strength calculations

    SciTech Connect

    Courant,E.D.

    2008-10-06

    In calculating the strengths of depolarizing resonances it may be convenient to reformulate the equations of spin motion in a coordinate system based on the actual trajectory of the particle, as introduced by Kondratenko, rather than the conventional one based on a reference orbit. It is shown that resonance strengths calculated by the conventional and the revised formalisms are identical. Resonances induced by radiofrequency dipoles or solenoids are also treated; with rf dipoles it is essential to consider not only the direct effect of the dipole but also the contribution from oscillations induced by it.

  5. High strength alloys

    DOEpatents

    Maziasz, Phillip James; Shingledecker, John Paul; Santella, Michael Leonard; Schneibel, Joachim Hugo; Sikka, Vinod Kumar; Vinegar, Harold J.; John, Randy Carl; Kim, Dong Sub

    2012-06-05

    High strength metal alloys are described herein. At least one composition of a metal alloy includes chromium, nickel, copper, manganese, silicon, niobium, tungsten and iron. System, methods, and heaters that include the high strength metal alloys are described herein. At least one heater system may include a canister at least partially made from material containing at least one of the metal alloys. At least one system for heating a subterranean formation may include a tublar that is at least partially made from a material containing at least one of the metal alloys.

  6. High strength alloys

    DOEpatents

    Maziasz, Phillip James [Oak Ridge, TN; Shingledecker, John Paul [Knoxville, TN; Santella, Michael Leonard [Knoxville, TN; Schneibel, Joachim Hugo [Knoxville, TN; Sikka, Vinod Kumar [Oak Ridge, TN; Vinegar, Harold J [Bellaire, TX; John, Randy Carl [Houston, TX; Kim, Dong Sub [Sugar Land, TX

    2010-08-31

    High strength metal alloys are described herein. At least one composition of a metal alloy includes chromium, nickel, copper, manganese, silicon, niobium, tungsten and iron. System, methods, and heaters that include the high strength metal alloys are described herein. At least one heater system may include a canister at least partially made from material containing at least one of the metal alloys. At least one system for heating a subterranean formation may include a tubular that is at least partially made from a material containing at least one of the metal alloys.

  7. Subcellular localization of rat CYP2E1 impacts metabolic efficiency toward common substrates.

    PubMed

    Hartman, Jessica H; Martin, H Cass; Caro, Andres A; Pearce, Amy R; Miller, Grover P

    2015-12-01

    Cytochrome P450 2E1 (CYP2E1) detoxifies or bioactivates many low molecular-weight compounds. Most knowledge about CYP2E1 activity relies on studies of the enzyme localized to endoplasmic reticulum (erCYP2E1); however, CYP2E1 undergoes transport to mitochondria (mtCYP2E1) and becomes metabolically active. We report the first comparison of in vitro steady-state kinetic profiles for erCYP2E1 and mtCYP2E1 oxidation of probe substrate 4-nitrophenol and pollutants styrene and aniline using subcellular fractions from rat liver. For all substrates, metabolic efficiency changed with substrate concentration for erCYP2E1 reflected in non-hyperbolic kinetic profiles but not for mtCYP2E1. Hyperbolic kinetic profiles for the mitochondrial enzyme were consistent with Michaelis-Menten mechanism in which metabolic efficiency was constant. By contrast, erCYP2E1 metabolism of 4-nitrophenol led to a loss of enzyme efficiency at high substrate concentrations when substrate inhibited the reaction. Similarly, aniline metabolism by erCYP2E1 demonstrated negative cooperativity as metabolic efficiency decreased with increasing substrate concentration. The opposite was observed for erCYP2E1 oxidation of styrene; the sigmoidal kinetic profile indicated increased efficiency at higher substrate concentrations. These mechanisms and CYP2E1 levels in mitochondria and endoplasmic reticulum were used to estimate the impact of CYP2E1 subcellular localization on metabolic flux of pollutants. Those models showed that erCYP2E1 mainly carries out aniline metabolism at all aniline concentrations. Conversely, mtCYP2E1 dominates styrene oxidation at low styrene concentrations and erCYP2E1 at higher concentrations. Taken together, subcellular localization of CYP2E1 results in distinctly different enzyme activities that could impact overall metabolic clearance and/or activation of substrates and thus impact the interpretation and prediction of toxicological outcomes. PMID:26463279

  8. High strength composites evaluation

    SciTech Connect

    Marten, S.M.

    1992-02-01

    A high-strength, thick-section, graphite/epoxy composite was identified. The purpose of this development effort was to evaluate candidate materials and provide LANL with engineering properties. Eight candidate materials (Samples 1000, 1100, 1200, 1300, 1400, 1500, 1600, and 1700) were chosen for evaluation. The Sample 1700 thermoplastic material was the strongest overall.

  9. The Strength of Vulnerability

    ERIC Educational Resources Information Center

    Gilliam, Bobby; Franklin, John Travis

    2004-01-01

    Many aspects of our work with at-risk children are spiritual by nature. A whole generation of at-risk children are crying out and asking hard questions. Although we certainly will not have all the answers, a shared experience of the very vulnerability of our human condition can turn this into a strength for us and our children. The authors propose…

  10. Suture anchor strength revisited.

    PubMed

    Barber, F A; Herbert, M A; Click, J N

    1996-02-01

    The rapid proliferation of suture anchors continues. Our prior report on the pullout strength of 14 different anchors is supplemented by a similar test conducted on 8 additional anchors. Comparative data on modes of failure and failure strengths (ultimate loads to failure) for these new devices are compared statistically with the previously tested anchors. In a fresh never-frozen porcine femur model, 10 samples of each of the additional anchors tested were threaded with stainless steel sutures and inserted into three different test areas (diaphyseal cortex, metaphyseal cortex, and a cancellous trough). Tensile stress parallel to the axis of insertion was applied at a rate of 12.5 mm/s by an Instron 1321 testing machine (Instron Corp, Canton, MA) until failure and mean anchor failure strengths calculated. The anchors tested were the Mitek G2 as a control, miniMitek, Mitek Superanchor, Mitek Rotator Cuff anchor (Mitek Products, Westwood, MA), Innovasive Devices Radial Osteal Compression device (Innovasive Devices, Hopkinton, MA), Arthrex Fastak (Arthrex Inc, Naples, FL), Arthrotek miniHarpoon (Arthrotek, Warsaw, IN), Orthopedic Biosystems PeBA 3 and PeBA 5 (Orthopedic Biosystems, Scottsdale, AZ), and AME 5.5 screw (American Medical Electronics, Richardson, TX). Failure mode (anchor pullout, suture eyelet cut out, or wire breakage) was generally consistent for each anchor type. The size of insertion hole is clinically important and each anchor's performance was evaluated as a function of its minor diameter or drill hole. For screw anchors, the larger the minor diameter of the screw, the higher the mean failure strengths in all three test areas (P = .001). However, larger drill holes for non-screw anchors resulted in lower mean failure strengths in cancellous bone (P = .03) and diaphyseal cortex (P < .005). PMID:8838726

  11. Structural models of the membrane anchors of envelope glycoproteins E1 and E2 from pestiviruses

    SciTech Connect

    Wang, Jimin Li, Yue; Modis, Yorgo

    2014-04-15

    The membrane anchors of viral envelope proteins play essential roles in cell entry. Recent crystal structures of the ectodomain of envelope protein E2 from a pestivirus suggest that E2 belongs to a novel structural class of membrane fusion machinery. Based on geometric constraints from the E2 structures, we generated atomic models of the E1 and E2 membrane anchors using computational approaches. The E1 anchor contains two amphipathic perimembrane helices and one transmembrane helix; the E2 anchor contains a short helical hairpin stabilized in the membrane by an arginine residue, similar to flaviviruses. A pair of histidine residues in the E2 ectodomain may participate in pH sensing. The proposed atomic models point to Cys987 in E2 as the site of disulfide bond linkage with E1 to form E1–E2 heterodimers. The membrane anchor models provide structural constraints for the disulfide bonding pattern and overall backbone conformation of the E1 ectodomain. - Highlights: • Structures of pestivirus E2 proteins impose constraints on E1, E2 membrane anchors. • Atomic models of the E1 and E2 membrane anchors were generated in silico. • A “snorkeling” arginine completes the short helical hairpin in the E2 membrane anchor. • Roles in pH sensing and E1–E2 disulfide bond formation are proposed for E1 residues. • Implications for E1 ectodomain structure and disulfide bonding pattern are discussed.

  12. Promoter-specific trans-activation by the adenovirus E1A12S product involves separate E1A domains.

    PubMed Central

    Kraus, V B; Moran, E; Nevins, J R

    1992-01-01

    Recent studies have shown that the adenovirus E1A12S product can trans-activate transcription by activating the transcription factor E2F. However, E2F cannot be the only target for the E1A12S product, since several cellular promoters have been found to be activated by the E1A12S protein even though they lack E2F sites. Indeed, we now show that activation of the hsp70 promoter by the E1A12S product requires the TATAA sequence. Moreover, activation of the hsp70 promoter requires the N-terminal domain of the E1A protein and does not require the conserved region 2 sequences which are required for the E2F-dependent activation of transcription. We conclude that the targeting of distinct transcription factors, leading to trans-activation of transcription of multiple promoters, involves distinct domains of the E1A proteins that are also required for oncogenic activity. Images PMID:1406628

  13. Associations of CYP2E1 rs2031920 and rs3813867 polymorphisms with colorectal cancer risk: a systemic review and meta-analysis.

    PubMed

    Peng, Hui; Xie, Shang-Kui; Huang, Mei-Jin; Ren, Dong-Lin

    2013-08-01

    Cytochrome P450 2E1 (CYP2E1) is a natural enzyme involved in the metabolic activation of many carcinogens, and the functional polymorphisms in the CYP2E1 gene might have impacts on colorectal cancer risk. Many studies were published to assess the associations of CYP2E1 rs2031920 and rs3813867 polymorphisms with colorectal cancer risk, but no consistent findings were reported. A systemic review and meta-analysis of eligible studies was performed to comprehensively assess the associations above. Odds ratios (ORs) with 95 % confidence interval (95 % CIs) were used to assess the strength of the associations. Seventeen studies from 15 publications with 17,082 individuals were finally included into this meta-analysis. Meta-analysis of the 13 studies on CYP2E1 rs2031920 polymorphism showed that there was a significant association between CYP2E1 rs2031920 polymorphism and colorectal cancer risk under two genetic models (c2 versus c1: OR = 1.19, 95 % CI 1.03-1.37, P = 0.022; c2c2/c2c1 versus c1c1: OR = 1.16, 95 % CI 1.00-1.35, P = 0.046). Meta-analysis of those four case-control studies on CYP2E1 rs3813867 polymorphism showed that there was no significant association between CYP2E1 rs3813867 polymorphism and colorectal cancer risk under all contrast models (c2 versus c1: OR = 0.96, 95 % CI 0.80-1.16, P = 0.672; c2c2 versus c1c1: OR = 1.26, 95 % CI 0.43-3.67, P = 0.672; c2c2/c1c2 versus c1c1: OR = 0.95, 95 % CI 0.78-1.16, P = 0.114; and c2c2 versus c1c2/c1c1: OR = 1.17, 95 % CI 0.41-3.36, P = 0.775). Therefore, the findings from this meta-analysis suggest that CYP2E1 rs2031920 polymorphism is associated with colorectal cancer risk, but CYP2E1 rs3813867 polymorphism is not associated with colorectal cancer risk. In addition, more well-designed studies with large sample size are needed to provide a more precise evaluation on the associations above. PMID:23595220

  14. Mechanism-based inactivation of human CYP2E1 by diethyldithocarbamate.

    PubMed

    Pratt-Hyatt, Matthew; Lin, Hsia-lien; Hollenberg, Paul F

    2010-12-01

    Although the ability of disulfiram to inactivate CYP2E1 has been known for more than 20 years, the mechanism has not yet been elucidated. A metabolite of disulfiram, diethyldithocarbamate (DDC), is converted by CYP2E1 to a reactive intermediate that subsequently inactivates the protein, leading to mechanism-based inactivation. Mass spectral analysis of the inactivated human 2E1 protein demonstrates that the inactivation is due to the formation of an adduct of the reactive metabolite of DDC with the apoprotein. These data, along with mass spectral analysis of a reactive intermediate trapped with GSH, indicate the involvement of a reactive intermediate with a molecular mass of 116 Da. Our results suggest that this binding involves formation of a disulfide bond with one of the eight cysteines in CYP2E1. The inactivation of wild-type CYP2E1 as well as two of its polymorphic mutants, CYP2E1*2 and CYP2E1*4, was also investigated. For wild-type CYP2E1, the K(I) was 12.2 μM and the k(inact) was 0.02 min(-1). The K(I) values for the two polymorphic mutants were 227.6 and 12.4 μM for CYP2E1.2 and CYP2E1.4, and the k(inact) values were 0.0061 and 0.0187 min(-1), respectively. These data indicate that DDC is a much less efficient inactivator of CYP2E1.2 than it is of either the wild-type or the CYP2E1.4 variant. PMID:20826547

  15. Adenovirus E1A proteins interact with the cellular YY1 transcription factor.

    PubMed Central

    Lewis, B A; Tullis, G; Seto, E; Horikoshi, N; Weinmann, R; Shenk, T

    1995-01-01

    The adenovirus 12S and 13S E1A proteins have been shown to relieve repression mediated by the cellular transcription factor YY1. The 13S E1A protein not only relieves repression but also activates transcription through YY1 binding sites. In this study, using a variety of in vivo and in vitro assays, we demonstrate that both E1A proteins can bind to YY1, although the 13S E1A protein binds more efficiently than the 12S E1A protein. Two domains on the E1A proteins interact with YY1: an amino-terminal sequence (residues 15 to 35) that is present in both E1A proteins and a domain that includes at least a portion of conserved region 3 (residues 140 to 188) that is present in the 13S but not the 12S E1A protein. Two domains on YY1 interact with E1A proteins: one is contained within residues 54 to 260, and the other is contained within the carboxy-terminal domain of YY1 (residues 332 to 414). Cotransfection of a plasmid expressing carboxy-terminal amino acids 332 to 414 of YY1 fused to the GAL4 DNA-binding domain can inhibit expression from a reporter construct with GAL4 DNA binding sites in its promoter, and inclusion of a third plasmid expressing E1A proteins can relieve the repression. Thus, we find a correlation between the ability of E1A to interact with the carboxy-terminal domain of YY1 and its ability to relieve repression caused by the carboxy-terminal domain of YY1. We propose that E1A proteins normally relieve YY1-mediated transcriptional repression by binding directly to the cellular transcription factor. PMID:7853498

  16. 26 CFR 1.1059(e)-1 - Non-pro rata redemptions.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Non-pro rata redemptions. 1.1059(e)-1 Section 1... (CONTINUED) INCOME TAXES Special Rules § 1.1059(e)-1 Non-pro rata redemptions. (a) In general. Section 1059(d... 1059(e)(1). For example, if a redemption of stock is not pro rata as to all shareholders, any...

  17. Methodology to assay CYP2E1 mixed function oxidase catalytic activity and its induction

    PubMed Central

    Cederbaum, Arthur I.

    2014-01-01

    The cytochrome P450 mixed function oxidase enzymes are the major catalysts involved in drug metabolism. There are many forms of P450. CYP2E1 metabolizes many toxicologically important compounds including ethanol and is active in generating reactive oxygen species. Since several of the contributions in the common theme series “Role of CYP2E1 and Oxidative/Nitrosative Stress in the Hepatotoxic Actions of Alcohol” discuss CYP2E1, this methodology review describes assays on how CYP2E1 catalytic activity and its induction by ethanol and other inducers can be measured using substrate probes such as the oxidation of para-nitrophenol to para-nitrocatechol and the oxidation of ethanol to acetaldehyde. Approaches to validate that a particular reaction e.g. oxidation of a drug or toxin is catalyzed by CYP2E1 or that induction of that reaction is due to induction of CYP2E1 are important and specific examples using inhibitors of CYP2E1, anti-CYP2E1 IgG or CYP2E1 knockout and knockin mice will be discussed. PMID:25454746

  18. Identification and characterization of multiple conserved nuclear localization signals within adenovirus E1A

    SciTech Connect

    Marshall, Kris S.; Cohen, Michael J.; Fonseca, Greg J.; Todorovic, Biljana; King, Cason R.; Yousef, Ahmed F.; Zhang, Zhiying; Mymryk, Joe S.

    2014-04-15

    The human adenovirus 5 (HAdV-5) E1A protein has a well defined canonical nuclear localization signal (NLS) located at its C-terminus. We used a genetic assay in the yeast Saccharomyces cerevisiae to demonstrate that the canonical NLS is present and functional in the E1A proteins of each of the six HAdV species. This assay also detects a previously described non-canonical NLS within conserved region 3 and a novel active NLS within the N-terminal/conserved region 1 portion of HAdV-5 E1A. These activities were also present in the E1A proteins of each of the other five HAdV species. These results demonstrate that, despite substantial differences in primary sequence, HAdV E1A proteins are remarkably consistent in that they contain one canonical and two non-canonical NLSs. By utilizing independent mechanisms, these multiple NLSs ensure nuclear localization of E1A in the infected cell. - Highlights: • HAdV E1A uses multiple mechanisms for nuclear import. • We identified an additional non-canonical NLS in the N-terminal/CR1 portion of E1A. • The new NLS does not contact importin-alpha directly. • All NLSs are functionally conserved in the E1A proteins of all 6 HAdV species.

  19. DNA-binding properties of the E1A-associated 300-kilodalton protein.

    PubMed Central

    Rikitake, Y; Moran, E

    1992-01-01

    One of the major E1A-associated cellular proteins is a 300-kDa product (p300) that binds to the N-terminal region of the E1A products. The p300 binding site is distinct from sequences involved in binding the retinoblastoma product and other E1A-associated cellular products such as p60-cyclin A and p107. p300 binding to E1A is linked genetically to the enhancer repression function of E1A and the other E1A-mediated gene-regulating functions as well as to the transforming functions of E1A. However, the biochemical properties of p300 have not yet been characterized. We report here that p300 has an intrinsic DNA-binding activity and shows a preferential affinity for specific DNA sequences. The sequences selectively bound by p300 are related to those of a series of enhancer elements that are recognized by NF-kappa B. The direct physical interaction of p300 with enhancer elements provides a biochemical basis for the genetic evidence linking the E1A-mediated enhancer repression function with the p300-binding activity of E1A. Images PMID:1534143

  20. Bone strength: current concepts.

    PubMed

    Turner, Charles H

    2006-04-01

    Bones serve several mechanical functions, including acoustic amplification in the middle ear, shielding vital organs from trauma, and serving as levers for muscles to contract against. Bone is a multiphase material made up of a tough collagenous matrix intermingled with rigid mineral crystals. The mineral gives bone its stiffness. Without sufficient mineralization, bones will plastically deform under load. Collagen provides toughness to bone making it less brittle so that it better resists fracture. Bone adapts to mechanical stresses largely by changing its size and shape, which are major determinants of its resistance to fracture. Tissue is added in regions of high mechanical stress providing an efficient means for improving bone strength. Experiments have shown that small additions of bone mineral density (BMD) (5-8%) caused by mechanical loading can improve bone strength by over 60% and extend bone fatigue life by 100-fold. Consequently, it is clear that bone tissue possesses a mechanosensing apparatus that directs osteogenesis to where it is most needed for improving bone strength. The biological processes involved in bone mechanotransduction are poorly understood and further investigation of the molecular mechanisms involved might uncover drug targets for osteoporosis. Several pathways are emerging from current research, including membrane ion channels, ATP signaling, second messengers, such as prostaglandins and nitric oxide, insulin-like growth factors, and Wnt signaling. PMID:16831941

  1. In vitro mineralization of MC3T3-E1 osteoblast-like cells on collagen/nano-hydroxyapatite scaffolds coated carbon/carbon composites.

    PubMed

    Cao, Sheng; Li, Hejun; Li, Kezhi; Lu, Jinhua; Zhang, Leilei

    2016-02-01

    Collagen/nano-hydroxyapatite (collagen/nHA) scaffolds were successfully prepared on carbon/carbon composites as bioactive films using the layer-by-layer coating method. Surface characterizations of collagen/nHA scaffolds were detected by scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Compressive strengths of the scaffolds were evaluated by a universal test machine. In vitro biological performances were determined using scaffolds seeded with MC3T3-E1 osteoblasts-like cells and cultured in mineralization medium for up to 21 days. In addition, cellular morphologies and several related gene expressions of MC3T3-E1 cells in the scaffolds were also evaluated. Chemical and morphological analysis showed that the scaffolds had uniform pore sizes and unified phase composition. Mechanical testing indicated that the collagen/nHA scaffolds had the highest compressive strength in 50% of strain condition when the proportion of collagen and nano-hydroxyapatite was 1:3. Cellular morphology observations and cytology tests indicated that MC3T3-E1 cells were adhered on these scaffolds and proliferated. SEM photographs and gene expressions showed that mineralized MC3T3-E1 cells and newly formed extra cellular matrix (ECM) filled up the pores of the scaffolds after the 3-week mineralization inducement. Nano-sized apatite particles were secreted from MC3T3-E1 cells and combined with the reconstructed ECM. Collectively, collagen/nHA scaffolds provided C/C composites with a biomimetic surface for cell adhesion, proliferation and mineralized extra cellular matrices formation. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 533-543, 2016. PMID:26476098

  2. Ubiquitin-dependent Proteasomal Degradation of Human Liver Cytochrome P450 2E1

    PubMed Central

    Wang, YongQiang; Guan, Shenheng; Acharya, Poulomi; Koop, Dennis R.; Liu, Yi; Liao, Mingxiang; Burlingame, Alma L.; Correia, Maria Almira

    2011-01-01

    Human liver CYP2E1 is a monotopic, endoplasmic reticulum-anchored cytochrome P450 responsible for the biotransformation of clinically relevant drugs, low molecular weight xenobiotics, carcinogens, and endogenous ketones. CYP2E1 substrate complexation converts it into a stable slow-turnover species degraded largely via autophagic lysosomal degradation. Substrate decomplexation/withdrawal results in a fast turnover CYP2E1 species, putatively generated through its futile oxidative cycling, that incurs endoplasmic reticulum-associated ubiquitin-dependent proteasomal degradation (UPD). CYP2E1 thus exhibits biphasic turnover in the mammalian liver. We now show upon heterologous expression of human CYP2E1 in Saccharomyces cerevisiae that its autophagic lysosomal degradation and UPD pathways are evolutionarily conserved, even though its potential for futile catalytic cycling is low due to its sluggish catalytic activity in yeast. This suggested that other factors (i.e. post-translational modifications or “degrons”) contribute to its UPD. Indeed, in cultured human hepatocytes, CYP2E1 is detectably ubiquitinated, and this is enhanced on its mechanism-based inactivation. Studies in Ubc7p and Ubc5p genetically deficient yeast strains versus corresponding isogenic wild types identified these ubiquitin-conjugating E2 enzymes as relevant to CYP2E1 UPD. Consistent with this, in vitro functional reconstitution analyses revealed that mammalian UBC7/gp78 and UbcH5a/CHIP E2-E3 ubiquitin ligases were capable of ubiquitinating CYP2E1, a process enhanced by protein kinase (PK) A and/or PKC inclusion. Inhibition of PKA or PKC blocked intracellular CYP2E1 ubiquitination and turnover. Here, through mass spectrometric analyses, we identify some CYP2E1 phosphorylation/ubiquitination sites in spatially associated clusters. We propose that these CYP2E1 phosphorylation clusters may serve to engage each E2-E3 ubiquitination complex in vitro and intracellularly. PMID:21209460

  3. Genetic polymorphism analysis of cytochrome P4502E1 (CYP2E1) in Chinese Han populations from four different geographic areas of Mainland China.

    PubMed

    Tang, Kefu; Li, Xin; Xing, Qinghe; Li, Weidong; Feng, Guoyin; He, Lin; Qin, Shengying

    2010-04-01

    CYP2E1 is one of a superfamily of enzymes that play a central role in activating and detoxifying many xenobiotics and endogenous compounds thought to be involved in the development of several human diseases. Among other factors, individual susceptibility to developing these pathologies relies on genetic polymorphisms, which are related to ethnic differences, since the frequency of mutant genotypes varies in different populations. The aim of this study was to investigate the genetic basis of CYP2E1 polymorphisms in the populations of four different geographical locations of China. Twenty-two different CYP2E1 polymorphisms, including six novel variants in promoter regions and a novel nonsense mutation, were identified. The frequencies of some polymorphisms and genotypes demonstrated significant differences among the four populations. Linkage disequilibrium analysis and tag SNP selection were performed. Haplotypes were analyzed within the selected tag SNPs. Tag SNP selection and haplotype distributions showed differences across the four populations. PMID:20100563

  4. Resolvin E1 inhibits dendritic cell migration in the skin and attenuates contact hypersensitivity responses.

    PubMed

    Sawada, Yu; Honda, Tetsuya; Hanakawa, Sho; Nakamizo, Satoshi; Murata, Teruasa; Ueharaguchi-Tanada, Yuri; Ono, Sachiko; Amano, Wataru; Nakajima, Saeko; Egawa, Gyohei; Tanizaki, Hideaki; Otsuka, Atsushi; Kitoh, Akihiko; Dainichi, Teruki; Ogawa, Narihito; Kobayashi, Yuichi; Yokomizo, Takehiko; Arita, Makoto; Nakamura, Motonobu; Miyachi, Yoshiki; Kabashima, Kenji

    2015-10-19

    Resolvin E1 (RvE1) is a lipid mediator derived from ω3 polyunsaturated fatty acids that exerts potent antiinflammatory roles in several murine models. The antiinflammatory mechanism of RvE1 in acquired immune responses has been attributed to attenuation of cytokine production by dendritic cells (DCs). In this study, we newly investigated the effect of RvE1 on DC motility using two-photon microscopy in a contact hypersensitivity (CHS) model and found that RvE1 impaired DC motility in the skin. In addition, RvE1 attenuated T cell priming in the draining lymph nodes and effector T cell activation in the skin, which led to the reduced skin inflammation in CHS. In contrast, leukotriene B4 (LTB4) induced actin filament reorganization in DCs and increased DC motility by activating Cdc42 and Rac1 via BLT1, which was abrogated by RvE1. Collectively, our results suggest that RvE1 attenuates cutaneous acquired immune responses by inhibiting cutaneous DC motility, possibly through LTB4-BLT1 signaling blockade. PMID:26438363

  5. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... these regulations apply? (a) This part applies to grants under section 419 of the Act (42 U.S.C. 285b-1... 42 Public Health 1 2011-10-01 2011-10-01 false To what programs do these regulations apply? 52e.1 Section 52e.1 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS...

  6. 26 CFR 1.263(e)-1 - Expenditures in connection with certain railroad rolling stock.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... by the Interstate Commerce Commission (49 CFR Part 1201), but only if (i) such unit exclusively moves... rolling stock. 1.263(e)-1 Section 1.263(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Not Deductible §...

  7. 26 CFR 1.263(e)-1 - Expenditures in connection with certain railroad rolling stock.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... by the Interstate Commerce Commission (49 CFR Part 1201), but only if (i) such unit exclusively moves... rolling stock. 1.263(e)-1 Section 1.263(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Not Deductible §...

  8. 26 CFR 1.263(e)-1 - Expenditures in connection with certain railroad rolling stock.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... by the Interstate Commerce Commission (49 CFR Part 1201), but only if (i) such unit exclusively moves... rolling stock. 1.263(e)-1 Section 1.263(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Not Deductible §...

  9. 26 CFR 1.263(e)-1 - Expenditures in connection with certain railroad rolling stock.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... by the Interstate Commerce Commission (49 CFR Part 1201), but only if (i) such unit exclusively moves... rolling stock. 1.263(e)-1 Section 1.263(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Not Deductible §...

  10. Nr2e1 Deficiency Augments Palmitate-Induced Oxidative Stress in Beta Cells

    PubMed Central

    Shi, Xiaoli; Deng, Haohua; Dai, Zhe; Xu, Yancheng; Xiong, Xiaokan; Ma, Pei; Cheng, Jing

    2016-01-01

    Nuclear receptor subfamily 2 group E member 1 (Nr2e1) has been regarded as an essential regulator of the growth of neural stem cells. However, its function elsewhere is unknown. In the present study, we generated Nr2e1 knockdown MIN6 cells and studied whether Nr2e1 knockdown affected basal beta cell functions such as proliferation, cell death, and insulin secretion. We showed that knockdown of Nr2e1 in MIN6 cells resulted in increased sensitivity to lipotoxicity, decreased proliferation, a partial G0/G1 cell-cycle arrest, and higher rates of apoptosis. Moreover, Nr2e1 deficiency exaggerates palmitate-induced impairment in insulin secretion. At the molecular level, Nr2e1 deficiency augments palmitate-induced oxidative stress. Nr2e1 deficiency also resulted in decreases in antioxidant enzymes and expression level of Nrf2. Together, this study indicated a potential protective effect of Nr2e1 on beta cells, which may serve as a target for the development of novel therapies for diabetes. PMID:26649147

  11. 26 CFR 1.1031(e)-1 - Exchange of livestock of different sexes.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 11 2012-04-01 2012-04-01 false Exchange of livestock of different sexes. 1.1031(e)-1 Section 1.1031(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... Exchange of livestock of different sexes. Section 1031(e) provides that livestock of different sexes...

  12. 26 CFR 1.1031(e)-1 - Exchange of livestock of different sexes.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 11 2010-04-01 2010-04-01 true Exchange of livestock of different sexes. 1.1031(e)-1 Section 1.1031(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... livestock of different sexes. Section 1031(e) provides that livestock of different sexes are not property...

  13. 26 CFR 1.1031(e)-1 - Exchange of livestock of different sexes.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 11 2014-04-01 2014-04-01 false Exchange of livestock of different sexes. 1.1031(e)-1 Section 1.1031(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... Exchange of livestock of different sexes. Section 1031(e) provides that livestock of different sexes...

  14. 26 CFR 1.1031(e)-1 - Exchange of livestock of different sexes.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 11 2011-04-01 2011-04-01 false Exchange of livestock of different sexes. 1.1031(e)-1 Section 1.1031(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... Exchange of livestock of different sexes. Section 1031(e) provides that livestock of different sexes...

  15. 26 CFR 1.1031(e)-1 - Exchange of livestock of different sexes.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 11 2013-04-01 2013-04-01 false Exchange of livestock of different sexes. 1.1031(e)-1 Section 1.1031(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... Exchange of livestock of different sexes. Section 1031(e) provides that livestock of different sexes...

  16. Identification of a novel E1A response element in the mouse c-fos promoter.

    PubMed Central

    Gedrich, R W; Engel, D A

    1995-01-01

    Transcriptional activation of the c-fos gene in mouse S49 cells by the adenovirus 243-amino-acid E1A protein depends on domains of E1A that are also required for transformation and that bind the cellular protein p300. Activation additionally depends on stimulation of endogenous cyclic AMP (cAMP)-dependent protein kinase by analogs or inducers of cAMP. Transient transfection assays were used to analyze the c-fos promoter for sequences that confer responsiveness to E1A. Linker substitution and point mutants revealed that transcriptional activation by E1A depended on a cAMP response element (CRE) located at -67 relative to the start site of transcription and a neighboring binding site for transcription factor YY1 located at -54. A 22-bp sequence containing the -67 CRE and the -54 YY1 site was sufficient to confer responsiveness to a minimal E1B promoter and was termed the c-fos E1A response element (ERE). Function of the c-fos ERE depended on both the CRE and the YY1 site, since mutation of either site resulted in a loss of responsiveness to E1A. These results imply a specific functional interaction between CRE-binding proteins, transcription factor YY1, and E1A in the regulation of the c-fos gene. PMID:7884879

  17. 26 CFR 1.263(e)-1 - Expenditures in connection with certain railroad rolling stock.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... extensions thereof) for filing the return for the taxable year of election. An election under section 263(e... by the Interstate Commerce Commission (49 CFR Part 1201), but only if (i) such unit exclusively moves... rolling stock. 1.263(e)-1 Section 1.263(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF...

  18. 26 CFR 1.509(e)-1 - Definition of gross investment income.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Definition of gross investment income. 1.509(e)-1...) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Private Foundations § 1.509(e)-1 Definition of gross investment income. For the distinction between gross receipts and gross investment income, see §...

  19. Secretion of Semliki Forest virus membrane glycoprotein E1 from Bacillus subtilis.

    PubMed

    Lundström, K; Palva, I; Kääriäinen, L; Garoff, H; Sarvas, M; Pettersson, R F

    1985-02-01

    The gene coding for the Semliki Forest virus (SFV) membrane protein E1 was joined to a secretion vector containing the promoter and signal sequence regions of the alpha-amylase gene from Bacillus amyloliquefaciens. To facilitate secretion, the regions coding for the N-terminal signal peptide (the 6K protein) and the C-terminal hydrophobic transmembrane domain of the E1 gene were deleted. After transformation into B. Subtilis, E1 was shown by immunoblotting to be expressed at a low level (about 0.5-1 mg/1). Contrary to what was expected, most of the E1 remained cell-associated. Deletion of a residual 7 C-terminal amino acids from the 6K region neither increased the level of expression nor significantly improved the secretion. Immunofluorescence microscopy of protoplasts prepared from B. subtilis cells expressing E1 suggested that the cell-associated E1 was located at the outer surface of the bacterial membrane. Addition of protease inhibitors to the culture medium somewhat increased the amount of extracellular E1, suggesting that proteolytic degradation of the foreign gene product may be one reason for the low level of expression. This conclusion was also supported by experiments carried out in Bacillus minicells, which indicated that the expression of the E1 gene in the absence of synthesis of bacterial proteases was about the same as that of alpha-amylase expressed from the cloned gene using the same promoter and signal sequence. PMID:3920841

  20. Subunit affinities and stoichiometries of the human papillomavirus type 11 E1:E2:DNA complex.

    PubMed

    Chao, S F; Rocque, W J; Daniel, S; Czyzyk, L E; Phelps, W C; Alexander, K A

    1999-04-01

    The association between the papillomavirus E1 and E2 proteins is an important regulatory interaction, imparting coordinated control of viral transcription and replication. Using fluorescence polarization, we have characterized the interactions between HPV-11 E1, HPV-11 E2, and DNA in solution at equilibrium. For these studies, two double-stranded fluorescein-labeled oligonucleotides were prepared. The first fluorescent oligonucleotide, designated Fl-E2BS and containing a single E2 binding-site palindrome (ACCGN6CGGT), was used to determine the affinity of E2 for its DNA binding site. HPV-11 E2 bound Fl-E2BS with an apparent Kd of 0.84 nM. Binding was saturable and consistent with a single class of noninteracting sites. The second oligonucleotide, designated Fl-E1E2BS, contained both E1 and E2 sites in sequence derived directly from the HPV-11 origin of replication. Under titration conditions identical to those used for Fl-E2BS, the E2 protein exhibited reduced affinity for Fl-E1E2BS (Kd > 100 nM). E1 binding to Fl-E1E2BS was of very low affinity. Addition of excess HPV-11 E1 to Fl-E1E2BS lowered the dissociation constant for the E2:Fl-E1E2BS interaction to 2 nM. This effect was not dependent upon ATP or magnesium ion. Fluorescence polarization and other data suggest formation of a complex containing six E1 molecules and a single dimer of E2 bound to a single Fl-E1E2BS oligonucleotide; E2 dissociation from the final complex did not occur. In summary, physical interaction between E1 and E2 increases the DNA binding affinity of each. The role of this energy coupling may be to promote origin-specific binding of both E1 and E2 to DNA. PMID:10194380

  1. Distinct Features of Cap Binding by eIF4E1b Proteins

    PubMed Central

    Kubacka, Dorota; Miguel, Ricardo Núñez; Minshall, Nicola; Darzynkiewicz, Edward; Standart, Nancy; Zuberek, Joanna

    2015-01-01

    eIF4E1b, closely related to the canonical translation initiation factor 4E (eIF4E1a), cap-binding protein is highly expressed in mouse, Xenopus and zebrafish oocytes. We have previously characterized eIF4E1b as a component of the CPEB mRNP translation repressor complex along with the eIF4E-binding protein 4E-Transporter, the Xp54/DDX6 RNA helicase and additional RNA-binding proteins. eIF4E1b exhibited only very weak interactions with m7GTP-Sepharose and, rather than binding eIF4G, interacted with 4E-T. Here we undertook a detailed examination of both Xenopus and human eIF4E1b interactions with cap analogues using fluorescence titration and homology modeling. The predicted structure of eIF4E1b maintains the α + β fold characteristic of eIF4E proteins and its cap-binding pocket is similarly arranged by critical amino acids: Trp56, Trp102, Glu103, Trp166, Arg112, Arg157 and Lys162 and residues of the C-terminal loop. However, we demonstrate that eIF4E1b is 3-fold less well able to bind the cap than eIF4E1a, both proteins being highly stimulated by methylation at N7 of guanine. Moreover, eIF4E1b proteins are distinguishable from eIF4E1a by a set of conserved amino acid substitutions, several of which are located near to cap-binding residues. Indeed, eIF4E1b possesses several distinct features, namely, enhancement of cap binding by a benzyl group at N7 position of guanine, a reduced response to increasing length of the phosphate chain and increased binding to a cap separated by a linker from Sepharose, suggesting differences in the arrangement of the protein's core. In agreement, mutagenesis of the amino acids differentiating eIF4E1b from eIF4E1a reduces cap binding by eIF4E1a 2-fold, demonstrating their role in modulating cap binding. PMID:25463438

  2. Distinct features of cap binding by eIF4E1b proteins.

    PubMed

    Kubacka, Dorota; Miguel, Ricardo Núñez; Minshall, Nicola; Darzynkiewicz, Edward; Standart, Nancy; Zuberek, Joanna

    2015-01-30

    eIF4E1b, closely related to the canonical translation initiation factor 4E (eIF4E1a), cap-binding protein is highly expressed in mouse, Xenopus and zebrafish oocytes. We have previously characterized eIF4E1b as a component of the CPEB mRNP translation repressor complex along with the eIF4E-binding protein 4E-Transporter, the Xp54/DDX6 RNA helicase and additional RNA-binding proteins. eIF4E1b exhibited only very weak interactions with m(7)GTP-Sepharose and, rather than binding eIF4G, interacted with 4E-T. Here we undertook a detailed examination of both Xenopus and human eIF4E1b interactions with cap analogues using fluorescence titration and homology modeling. The predicted structure of eIF4E1b maintains the α+β fold characteristic of eIF4E proteins and its cap-binding pocket is similarly arranged by critical amino acids: Trp56, Trp102, Glu103, Trp166, Arg112, Arg157 and Lys162 and residues of the C-terminal loop. However, we demonstrate that eIF4E1b is 3-fold less well able to bind the cap than eIF4E1a, both proteins being highly stimulated by methylation at N(7) of guanine. Moreover, eIF4E1b proteins are distinguishable from eIF4E1a by a set of conserved amino acid substitutions, several of which are located near to cap-binding residues. Indeed, eIF4E1b possesses several distinct features, namely, enhancement of cap binding by a benzyl group at N(7) position of guanine, a reduced response to increasing length of the phosphate chain and increased binding to a cap separated by a linker from Sepharose, suggesting differences in the arrangement of the protein's core. In agreement, mutagenesis of the amino acids differentiating eIF4E1b from eIF4E1a reduces cap binding by eIF4E1a 2-fold, demonstrating their role in modulating cap binding. PMID:25463438

  3. Oxidative stress mediated toxicity exerted by ethanol-inducible CYP2E1

    SciTech Connect

    Wu Defeng; Cederbaum, Arthur I. . E-mail: arthur.cederbaum@mssm.edu

    2005-09-01

    Induction of CYP2E1 by ethanol is one of the central pathways by which ethanol generates a state of oxidative stress in hepatocytes. To study the biochemical and toxicological actions of CYP2E1, our laboratory established HepG2 cell lines which constitutively overexpress CYP2E1 and characterized these cells with respect to ethanol toxicity. Addition of ethanol or an unsaturated fatty acid such as arachidonic acid or iron was toxic to the CYP2E1-expressing cells but not control cells. This toxicity was associated with elevated lipid peroxidation and could be prevented by antioxidants and inhibitors of CYP2E1. Apoptosis occurred in the CYP2E1-expressing cells exposed to ethanol, arachidonic acid, or iron. Removal of GSH caused a loss of viability in the CYP2E1-expressing cells even in the absence of added toxin or pro-oxidant. This was associated with mitochondrial damage and decreased mitochondrial membrane potential. Low concentrations of iron and arachidonic acid synergistically interacted with CYP2E1 to produce cell toxicity, suggesting these nutrients may act as priming or sensitizing agents to alcohol-induced liver injury. Surprisingly, CYP2E1-expressing cells had elevated GSH levels, due to transcriptional activation of glutamate cysteine ligase. Similarly, levels of catalase, alpha-, and microsomal glutathione transferase were also increased, suggesting that upregulation of these antioxidant genes may reflect an adaptive mechanism to remove CYP2E1-derived oxidants. Using co-cultures, interaction between CYP2E1-derived diffusible mediators to activate collagen production in hepatic stellate cells was found. While it is likely that several mechanisms contribute to alcohol-induced liver injury, the linkage between CYP2E1-dependent oxidative stress, mitochondrial injury, stellate cell activation, and GSH homeostasis may contribute to the toxic action of ethanol on the liver. HepG2 cell lines overexpressing CYP2E1 may be a valuable model to characterize the biochemical and toxicological properties of CYP2E1.

  4. Determination of polymyxin E1 in rat plasma by high-performance liquid chromatography.

    PubMed

    Gmur, Dennis J; Bredl, Charles R; Steele, Sharon J; Cai, Shaopei; VanDevanter, Donald R; Nardella, Pasqua A

    2003-06-15

    A precise and accurate HPLC assay for polymyxin E(1) in rat and dog plasma has been validated. Samples and standards are extracted from plasma with a 96-well C(8) extraction disk plate. Sample extracts are derivatized with dansyl chloride, and polymyxin E(1) derivative is quantitated on a C(8) column by HPLC with fluorescence detection. The assay is linear in the range of 0.050-5.00 micro g/ml for polymyxin E(1). The precision and accuracy of polymyxin E(1) plasma assay was well within the recommended limits set in the FDA Guidance for Bioanalytical Method Validation. Polymyxin E(1) stability in rat and dog plasma for 24 h at room temperature and through three freeze-thaw cycles was demonstrated. PMID:12742127

  5. New protein vector ApE1 for targeted delivery of anticancer drugs.

    PubMed

    Pozdniakova, N V; Gorokhovets, N V; Gukasova, N V; Bereznikova, A V; Severin, E S

    2012-01-01

    A new chimeric gene ApE1 encoding the receptor-binding domain of the human alpha-fetoprotein fused to a sequence of 22 glutamic acid residues was constructed. A new bacterial producer strain E. coli SHExT7 ApE1 was selected for ApE1 production in a soluble state. A simplified method was developed to purify ApE1 from bacterial biomass. It was shown that the new vector protein selectively interacts with AFP receptors on the tumor cell surface and can be efficiently accumulated in tumor cells. In addition, ApE1 was shown to be stable in storage and during its chemical modification. An increased number of carboxyl groups in the molecule allows the production of cytotoxic compound conjugates with higher drug-loading capacity and enhanced tumor targeting potential. PMID:22649278

  6. New Protein Vector ApE1 for Targeted Delivery of Anticancer Drugs

    PubMed Central

    Pozdniakova, N. V.; Gorokhovets, N. V.; Gukasova, N. V.; Bereznikova, A. V.; Severin, E. S.

    2012-01-01

    A new chimeric gene ApE1 encoding the receptor-binding domain of the human alpha-fetoprotein fused to a sequence of 22 glutamic acid residues was constructed. A new bacterial producer strain E. coli SHExT7 ApE1 was selected for ApE1 production in a soluble state. A simplified method was developed to purify ApE1 from bacterial biomass. It was shown that the new vector protein selectively interacts with AFP receptors on the tumor cell surface and can be efficiently accumulated in tumor cells. In addition, ApE1 was shown to be stable in storage and during its chemical modification. An increased number of carboxyl groups in the molecule allows the production of cytotoxic compound conjugates with higher drug-loading capacity and enhanced tumor targeting potential. PMID:22649278

  7. Prepubescent Strength Training. Some Considerations.

    ERIC Educational Resources Information Center

    Priest, Joe W.; Holshouser, Richard S.

    1987-01-01

    Under the careful supervision of a trained fitness professional, the benefits of prepubescent strength training (improved strength, power, muscular endurance, bone density) outweigh the risks (acute and chronic musculoskeletal injuries). (CB)

  8. Role of CYP2E1 in thioacetamide-induced mouse hepatotoxicity

    SciTech Connect

    Kang, Jin Seok; Wanibuchi, Hideki; Morimura, Keiichirou; Wongpoomchai, Rawiwan; Chusiri, Yaowares; Gonzalez, Frank J.; Fukushima, Shoji

    2008-05-01

    Previous experiments showed that treatment of mice and rats with thioacetamide (TAA) induced liver cell damage, fibrosis and/or cirrhosis, associated with increased oxidative stress and activation of hepatic stellate cells. Some experiments suggest that CYP2E1 may be involved in the metabolic activation of TAA. However, there is no direct evidence on the role of CYP2E1 in TAA-mediated hepatotoxicity. To clarify this, TAA-induced hepatotoxicity was investigated using Cyp2e1-null mice. Male wild-type and Cyp2e1-null mice were treated with TAA (200 mg/kg of body weight, single, i.p.) at 6 weeks of age, and hepatotoxicity examined 24 and 48 h after TAA treatment. Relative liver weights of Cyp2e1-null mice were significantly different at 24 h compared to wild-type mice (p < 0.01). Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH) in Cyp2e1-null mice were significantly different at both time points compared to wild-type mice (p < 0.01). Histopathological examination showed Cyp2e1-null mice represented no hepatototoxic lesions, in clear contrast to severe centriobular necrosis, inflammation and hemorrhage at both time points in wild-type mice. Marked lipid peroxidation was also only limited to wild-type mice (p < 0.01). Similarly, TNF-{alpha}, IL-6 and glutathione peroxidase mRNA expression in Cyp2e1-null mice did not significantly differ from the control levels, contrasting with the marked alteration in wild-type mice (p < 0.01). Western blot analysis further revealed no increase in iNOS expression in Cyp2e1-null mice. These results reveal that CYP2E1 mediates TAA-induced hepatotoxicity in wild-type mice as a result of increased oxidative stress.

  9. Adenovirus E1A proteins inhibit activation of transcription by p53.

    PubMed Central

    Steegenga, W T; van Laar, T; Riteco, N; Mandarino, A; Shvarts, A; van der Eb, A J; Jochemsen, A G

    1996-01-01

    p53 stimulates the transcription of a number of genes, such as MDM2, Waf1, and GADD45. We and others have shown previously that this activity of p53 can be inhibited by adenovirus type 2 or 12 large E1B proteins. Here we show that the adenovirus E1A proteins also can repress the stimulation of transcription by p53, both in transient transfections and in stably transfected cell lines. The inhibition by E1A occurs without a significant effect on the DNA-binding capacity of p53. Furthermore, the activity of a fusion protein containing the N-terminal part of p53 linked to the GAL4 DNA-binding domain can be suppressed by E1A. This indicates that E1A affects the transcription activation domain of p53, although tryptic phosphopeptide mapping revealed that the level of phosphorylation of this domain does not change significantly in E1A-expressing cell lines. Gel filtration studies, however, showed p53 to be present in complexes of increased molecular weight as a result of E1A expression. Apparently, E1A can cause increased homo- or hetero-oligomerization of p53, which might result in the inactivation of the transcription activation domain of p53. Additionally, we found that transfectants stably expressing E1A have lost the ability to arrest in G1 after DNA damage, indicating that E1A can abolish the normal biological function of p53. PMID:8628276

  10. Architecture of the hepatitis C virus E1 glycoprotein transmembrane domain studied by NMR.

    PubMed

    Zazrin, Hadas; Shaked, Hadassa; Chill, Jordan H

    2014-03-01

    Oligomerization of hepatitis C viral envelope proteins E1 and E2 is essential to virus fusion and assembly. Although interactions within the transmembrane (TM) domains of these glycoproteins have proven contributions to the E1/E2 heterodimerization process and consequent infectivity, there is little structural information on this entry mechanism. Here, as a first step towards our long-term goal of understanding the interaction between E1 and E2 TM-domains, we have expressed, purified and characterized E1-TM using structural biomolecular NMR methods. An MBP-fusion expression system yielded sufficient quantities of pure E1-TM, which was solubilized in two membrane-mimicking environments, SDS- and LPPG-micelles, affording samples amenable to NMR studies. Triple resonance assignment experiments and relaxation measurements provided information on the secondary structure and global fold of E1-TM in these environments. In SDS micelles E1-TM adopts a helical conformation, with helical stretches at residues 354-363 and 371-379 separated by a more flexible segment of residues 364-370. In LPPG micelles a helical conformation was observed for residues 354-377 with greater flexibility in the 366-367 dyad, suggesting LPPG provides a more native environment for the peptide. Replacement of key positively charged residue K370 with an alanine did not affect the secondary structure of E1-TM but did change the relative positioning within the micelle of the two helices. These results lay the foundation for structure determination of E1-TM and a molecular understanding of how E1-TM flexibility enhances its interaction with E2-TM during heterodimerization and membrane fusion. PMID:24192053

  11. Folding of Hepatitis C Virus E1 Glycoprotein in a Cell-Free System

    PubMed Central

    Merola, Marcello; Brazzoli, Michela; Cocchiarella, Fabienne; Heile, Jens M.; Helenius, Ari; Weiner, Amy J.; Houghton, Michael; Abrignani, Sergio

    2001-01-01

    The hepatitis C virus (HCV) envelope proteins, E1 and E2, form noncovalent heterodimers and are leading candidate antigens for a vaccine against HCV. Studies in mammalian cell expression systems have focused primarily on E2 and its folding, whereas knowledge of E1 folding remains fragmentary. We used a cell-free in vitro translation system to study E1 folding and asked whether the flanking proteins, Core and E2, influence this process. We translated the polyprotein precursor, in which the Core is N-terminal to E1, and E2 is C-terminal, and found that when the core protein was present, oxidation of E1 was a slow, E2-independent process. The half-time for E1 oxidation was about 5 h in the presence or absence of E2. In contrast with previous reports, analysis of three constructs of different lengths revealed that the E2 glycoprotein undergoes slow oxidation as well. Unfolded or partially folded E1 bound to the endoplasmic reticulum chaperones calnexin and (with lower efficiency) calreticulin, whereas no binding to BiP/GRP78 or GRP94 could be detected. Release from calnexin and calreticulin was used to assess formation of mature E1. When E1 was expressed in the absence of Core and E2, its oxidation was impaired. We conclude that E1 folding is a process that is affected not only by E2, as previously shown, but also by the Core. The folding of viral proteins can thus depend on complex interactions between neighboring proteins within the polyprotein precursor. PMID:11602760

  12. Molecular mechanism of trichloroethylene-induced hepatotoxicity mediated by CYP2E1

    SciTech Connect

    Ramdhan, Doni Hikmat; Kamijima, Michihiro; Yamada, Naoyasu; Ito, Yuki; Yanagiba, Yukie; Nakamura, Daichi; Okamura, Ai; Ichihara, Gaku; Aoyama, Toshifumi; Gonzalez, Frank J.; Nakajima, Tamie

    2008-09-15

    Cytochrome P450 (CYP) 2E1 was suggested to be the major enzyme involved in trichloroethylene (TRI) metabolism and TRI-induced hepatotoxicity, although the latter molecular mechanism is not fully understood. The involvement of CYP2E1 in TRI-induced hepatotoxicity and its underlying molecular mechanism were studied by comparing hepatotoxicity in cyp2e1{sup +/+} and cyp2e1{sup -/-} mice. The mice were exposed by inhalation to 0 (control), 1000, or 2000 ppm of TRI for 8 h a day, for 7 days, and TRI-hepatotoxicity was assessed by measuring plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities and histopathology. Urinary metabolites of trichloroethanol and trichloroacetic acid (TCA) were considerably greater in cyp2e1{sup +/+} compared to cyp2e1{sup -/-} mice, suggesting that CYP2E1 is the major P450 involved in the formation of these metabolites. Consistent with elevated plasma ALT and AST activities, cyp2e1{sup +/+} mice in the 2000 ppm group showed histopathological inflammation. TRI significantly upregulated PPAR{alpha}, which might function to inhibit NF{kappa}B p50 and p65 signalling. In addition, TRI-induced NF{kappa}B p52 mRNA, and significantly positive correlation between NF{kappa}B p52 mRNA expression and plasma ALT activity levels were observed, suggesting the involvement of p52 in liver inflammation. Taken together, the current study directly demonstrates that CYP2E1 was the major P450 involved in the first step of the TRI metabolism, and the metabolites produced may have two opposing roles: one inducing hepatotoxicity and the other protecting against the toxicity. Intermediate metabolite(s) from TRI to chloral hydrate produced by CYP2E1-mediated oxidation may be involved in the former, and TCA in the latter.

  13. Effects of Adenovirus Type 5 E1A Isoforms on Viral Replication in Arrested Human Cells

    PubMed Central

    Radko, Sandi; Jung, Richard; Olanubi, Oladunni; Pelka, Peter

    2015-01-01

    Human adenovirus has evolved to infect and replicate in terminally differentiated human epithelial cells, predominantly those within the airway, the gut, or the eye. To overcome the block to viral DNA replication present in these cells, the virus expresses the Early 1A proteins (E1A). These immediate early proteins drive cells into S-phase and induce expression of all other viral early genes. During infection, several E1A isoforms are expressed with proteins of 289, 243, 217, 171, and 55 residues being present for human adenovirus type 5. Here we examine the contribution that the two largest E1A isoforms make to the viral life cycle in growth-arrested normal human fibroblasts. Viruses that express E1A289R were found to replicate better than those that do not express this isoform. Importantly, induction of several viral genes was delayed in a virus expressing E1A243R, with several viral structural proteins undetectable by western blot. We also highlight the changes in E1A isoforms detected during the course of viral infection. Furthermore, we show that viral DNA replication occurs more efficiently, leading to higher number of viral genomes in cells infected with viruses that express E1A289R. Finally, induction of S-phase specific genes differs between viruses expressing different E1A isoforms, with those having E1A289R leading to, generally, earlier activation of these genes. Overall, we provide an overview of adenovirus replication using modern molecular biology approaches and further insights into the contribution that E1A isoforms make to the life cycle of human adenovirus in arrested human fibroblasts. PMID:26448631

  14. Strength Training and Children's Health.

    ERIC Educational Resources Information Center

    Faigenbaum, Avery D.

    2001-01-01

    Provides an overview of the potential health benefits of strength training for children, discussing the role of strength training in preventing sports-related injuries and highlighting design considerations for such programs. The focus is on musculoskeletal adaptations to strength training that are observable in healthy children. Guidelines for…

  15. Strength Training for Young Athletes.

    ERIC Educational Resources Information Center

    Kraemer, William J.; Fleck, Steven J.

    This guide is designed to serve as a resource for developing strength training programs for children. Chapter 1 uses research findings to explain why strength training is appropriate for children. Chapter 2 explains some of the important physiological concepts involved in children's growth and development as they apply to developing strength

  16. Strength Development for Young Adolescents

    ERIC Educational Resources Information Center

    McDaniel, Larry W.; Jackson, Allen; Gaudet, Laura

    2009-01-01

    Participation in strength training is important for older children or young adolescences who wish to improve fitness or participate in sports. When designing strength training programs for our youth this age group is immature anatomically, physiologically, and psychologically. For the younger or inexperienced group the strength training activities…

  17. Strength of Rewelded Inconel 718

    NASA Technical Reports Server (NTRS)

    Bayless, E.; Lovoy, C. V.; Mcllwain, M. C.; Munafo, P.

    1982-01-01

    Inconel 718, nickel-based alloy used extensively for high-temperature structural service, welded repeatedly without detriment to its strength. According to NASA report, tests show 12 repairs on same weld joint do not adversely affect ultimate tensile strenth, yield strength, fatigue strength, metallurgical grain structures, or ability of weld joint to respond to post weld heat treatments.

  18. Strength Training and Children's Health.

    ERIC Educational Resources Information Center

    Faigenbaum, Avery D.

    2001-01-01

    Provides an overview of the potential health benefits of strength training for children, discussing the role of strength training in preventing sports-related injuries and highlighting design considerations for such programs. The focus is on musculoskeletal adaptations to strength training that are observable in healthy children. Guidelines for

  19. The fracture strength and frictional strength of Weber Sandstone

    USGS Publications Warehouse

    Byerlee, J.D.

    1975-01-01

    The fracture strength and frictional strength of Weber Sandstone have been measured as a function of confining pressure and pore pressure. Both the fracture strength and the frictional strength obey the law of effective stress, that is, the strength is determined not by the confining pressure alone but by the difference between the confining pressure and the pore pressure. The fracture strength of the rock varies by as much as 20 per cent depending on the cement between the grains, but the frictional strength is independent of lithology. Over the range 0 2 kb, ??=0??5 + 0??6??n. This relationship also holds for other rocks such as gabbro, dunite, serpentinite, granite and limestone. ?? 1975.

  20. Initial steps of colicin E1 import across the outer membrane of Escherichia coli.

    PubMed

    Masi, Muriel; Vuong, Phu; Humbard, Matthew; Malone, Karen; Misra, Rajeev

    2007-04-01

    Data suggest a two-receptor model for colicin E1 (ColE1) translocation across the outer membrane of Escherichia coli. ColE1 initially binds to the vitamin B(12) receptor BtuB and then translocates through the TolC channel-tunnel, presumably in a mostly unfolded state. Here, we studied the early events in the import of ColE1. Using in vivo approaches, we show that ColE1 is cleaved when added to whole cells. This cleavage requires the presence of the receptor BtuB and the protease OmpT, but not that of TolC. Strains expressing OmpT cleaved ColE1 at K84 and K95 in the N-terminal translocation domain, leading to the removal of the TolQA box, which is essential for ColE1's cytotoxicity. Supported by additional in vivo data, this suggests that a function of OmpT is to degrade colicin at the cell surface and thus protect sensitive E. coli cells from infection by E colicins. A genetic strategy for isolating tolC mutations that confer resistance to ColE1, without affecting other TolC functions, is also described. We provide further in vivo evidence of the multistep interaction between TolC and ColE1 by using cross-linking followed by copurification via histidine-tagged TolC. First, secondary binding of ColE1 to TolC is dependent on primary binding to BtuB. Second, alterations to a residue in the TolC channel interfere with the translocation of ColE1 across the TolC pore rather than with the binding of ColE1 to TolC. In contrast, a substitution at a residue exposed on the cell surface abolishes both binding and translocation of ColE1. PMID:17277071

  1. Initial Steps of Colicin E1 Import across the Outer Membrane of Escherichia coli▿

    PubMed Central

    Masi, Muriel; Vuong, Phu; Humbard, Matthew; Malone, Karen; Misra, Rajeev

    2007-01-01

    Data suggest a two-receptor model for colicin E1 (ColE1) translocation across the outer membrane of Escherichia coli. ColE1 initially binds to the vitamin B12 receptor BtuB and then translocates through the TolC channel-tunnel, presumably in a mostly unfolded state. Here, we studied the early events in the import of ColE1. Using in vivo approaches, we show that ColE1 is cleaved when added to whole cells. This cleavage requires the presence of the receptor BtuB and the protease OmpT, but not that of TolC. Strains expressing OmpT cleaved ColE1 at K84 and K95 in the N-terminal translocation domain, leading to the removal of the TolQA box, which is essential for ColE1's cytotoxicity. Supported by additional in vivo data, this suggests that a function of OmpT is to degrade colicin at the cell surface and thus protect sensitive E. coli cells from infection by E colicins. A genetic strategy for isolating tolC mutations that confer resistance to ColE1, without affecting other TolC functions, is also described. We provide further in vivo evidence of the multistep interaction between TolC and ColE1 by using cross-linking followed by copurification via histidine-tagged TolC. First, secondary binding of ColE1 to TolC is dependent on primary binding to BtuB. Second, alterations to a residue in the TolC channel interfere with the translocation of ColE1 across the TolC pore rather than with the binding of ColE1 to TolC. In contrast, a substitution at a residue exposed on the cell surface abolishes both binding and translocation of ColE1. PMID:17277071

  2. Effect of status epilepticus and antiepileptic drugs on CYP2E1 brain expression

    PubMed Central

    Boussadia, Badreddine; Ghosh, Chaitali; Plaud, Clement; Pascussi, Jean-Marc; deBock, Frederic; Rousset, Marie-Claude; Janigro, Damir; Marchi, Nicola

    2014-01-01

    P450 metabolic enzymes are expressed in the human and rodent brain. Recent data support their involvement in the pathophysiology of epilepsy. However, the determinants of metabolic enzyme expression in the epileptic brain are unclear. We tested the hypothesis that status epilepticus or exposure to phenytoin or phenobarbital affects brain expression of the metabolic enzyme CYP2E1. Status epilepticus (SE) was induced in C57BL/6J mice by systemic kainic acid. Brain CYP2E1 expression was evaluated 18–24 hours after SE by immunohistochemistry. Co-localization with NEUN, GFAP and CD31 was determined by confocal microscopy. The effect of phenytoin, carbamazepine and phenobarbital on CYP2E1 expression was evaluated in vivo or by using organotypic hippocampal cultures in vitro. CYP2E1 expression was investigated in brain resections from a cohort of drug resistant epileptic brain resections and human endothelial cultures (EPI-EC). Immunohistochemistry showed an increase of CYP2E1 expression limited to hippocampal CA2/3 and hilar neurons after severe SE in mice. CYP2E1 expression was also observed at the astrocyte-vascular interface. Analysis of human brain specimens revealed CYP2E1 expression in neurons and vascular endothelial cells (EC). CYP2E1 was expressed in cultured human EC and over-expressed by EPI-EC. When analyzing the effect of drug exposure on CYP2E1 expression we found that, in vivo or in vitro, ethanol increased CYP2E1 levels in the brain and liver. Treatment with phenytoin induced localized CYP2E1 expression in the brain whereas no significant effects were exerted by carbamazepine or phenobarbital. Our data indicate that the effect of acute SE on brain CYP2E1 expression is localized and cell specific. Exposure to selected anti-epileptic drugs could play a role in determining CYP2E1 brain expression. Additional investigation is required to fully reproduce the culprits of P450 enzyme expression as observed in the human epileptic brain. PMID:25280786

  3. Alcohol steatosis and cytotoxicity: the role of cytochrome P4502E1 and autophagy.

    PubMed

    Wu, Defeng; Wang, Xiaodong; Zhou, Richard; Yang, Lili; Cederbaum, Arthur I

    2012-09-15

    The goal of the current study was to evaluate whether CYP2E1 plays a role in binge-ethanol induced steatosis and if autophagy impacts CYP2E1-mediated hepatotoxicity, oxidative stress and fatty liver formation produced by ethanol. Wild type (WT), CYP2E1 knockin (KI) and CYP2E1 knockout (KO) mice were gavaged with 3g/kg body wt ethanol twice a day for four days. This treatment caused fatty liver, elevation of CYP2E1 and oxidative stress in WT and KI mice but not KO mice. Autophagy was impaired in ethanol-treated KI mice compared to KO mice as reflected by a decline in the LC3-II/LC3-I ratio and lower total LC-3 and Beclin-1 levels coupled to increases in P62, pAKT/AKT and mTOR. Inhibition of macroautophagy by administration of 3-methyladenine enhanced the binge ethanol hepatotoxicity, steatosis and oxidant stress in CYP2E1 KI, but not CYP2E1 KO mice. Stimulation of autophagy by rapamycin blunted the elevated steatosis produced by binge ethanol. Treatment of HepG2 E47 cells which express CYP2E1 with 100mM ethanol for 8 days increased fat accumulation and oxidant stress but decreased autophagy. Ethanol had no effect on these reactions in HepG2 C34 cells which do not express CYP2E1. Inhibition of autophagy elevated ethanol toxicity, lipid accumulation and oxidant stress in the E47, but not C34 cells. The antioxidant N-acetylcysteine, and CYP2E1 inhibitor chlormethiazole blunted these effects of ethanol. These results indicate that CYP2E1 plays an important role in binge ethanol-induced fatty liver. We propose that CYP2E1-derived reactive oxygen species inhibit autophagy, which subsequently causes accumulation of lipid droplets. Inhibition of autophagy promotes binge ethanol induced hepatotoxicity, steatosis and oxidant stress via CYP2E1. PMID:22819980

  4. Cytochrome P450 2E1 (CYP2E1) regulates the response to oxidative stress and migration of breast cancer cells

    PubMed Central

    2013-01-01

    Introduction The cytochrome P450 (CYP) enzymes are a class of heme-containing enzymes involved in phase I metabolism of a large number of xenobiotics. The CYP family member CYP2E1 metabolises many xenobiotics and pro-carcinogens, it is not just expressed in the liver but also in many other tissues such as the kidney, the lung, the brain, the gastrointestinal tract and the breast tissue. It is induced in several pathological conditions including cancer, obesity, and type II diabetes implying that this enzyme is implicated in other biological processes beyond its role in phase I metabolism. Despite the detailed description of the role of CYP2E1 in the liver, its functions in other tissues have not been extensively studied. In this study, we investigated the functional significance of CYP2E1 in breast carcinogenesis. Methods Cellular levels of reactive oxygen species (ROS) were measured by H2DCFDA (2 2.9.2 2′,7′-dichlorodihydrofluorescein diacetate) staining and autophagy was assessed by tracing the cellular levels of autophagy markers using western blot assays. The endoplasmic reticulum stress and the unfolded protein response (UPR) were detected by luciferase assays reflecting the splicing of mRNA encoding the X-box binding protein 1 (XBP1) transcription factor and cell migration was evaluated using the scratch wound assay. Gene expression was recorded with standard transcription assays including luciferase reporter and chromatin immunoprecipitation. Results Ectopic expression of CYP2E1 induced ROS generation, affected autophagy, stimulated endoplasmic reticulum stress and inhibited migration in breast cancer cells with different metastatic potential and p53 status. Furthermore, evidence is presented indicating that CYP2E1 gene expression is under the transcriptional control of the p53 tumor suppressor. Conclusions These results support the notion that CYP2E1 exerts an important role in mammary carcinogenesis, provide a potential link between ethanol metabolism and breast cancer and suggest that progression, and metastasis, of advanced stages of breast cancer can be modulated by induction of CYP2E1 activity. PMID:24207099

  5. Intranuclear location of the adenovirus type 5 E1B 55-kilodalton protein.

    PubMed Central

    Smiley, J K; Young, M A; Flint, S J

    1990-01-01

    The intracellular location of the adenovirus type 5 E1B 55-kilodalton (kDa) protein, particularly the question of whether it is associated with nuclear pore complexes, was examined. Fractionation of adenovirus type 5-infected HeLa cell nuclei by an established procedure (N. Dwyer and G. Blobel, J. Cell. Biol. 70:581-591, 1976) yielded one population of E1B 55-kDa protein molecules released by digestion of nuclei with RNase A and a second population recovered in the pore complex-lamina fraction. Free and E1B 55-kDa protein-bound forms of the E4 34-kDa protein (P. Sarnow, C. A. Sullivan, and A. J. Levine, Virology 120:387-394, 1982) were largely recovered in the pore complex-lamina fraction. Nevertheless, the association of E1B 55-kDa protein molecules with this nuclear envelope fraction did not depend on interaction of the E1B 55-kDa protein with the E4 34-kDa protein. Comparison of the immunofluorescence patterns observed with antibodies recognizing the E1B 55-kDa protein or cellular pore complex proteins and of the behavior of these viral and cellular proteins during in situ fractionation suggests that the E1B 55-kDa protein does not become intimately or stably associated with pore complexes in adenovirus-infected cells. Images PMID:2143545

  6. Evidence for a tissue-specific induction of cutaneous CYP2E1 by dexamethasone.

    PubMed

    Sampol, E; Mirrione, A; Villard, P H; Piccerelle, P; Scoma, H; Berbis, P; Barra, Y; Durand, A; Lacarelle, B

    1997-06-27

    We studied in mouse the effect of topical application of dexamethasone or salicylic acid, on CYP2E1 and CYP3A expression (proteins and/or mRNA) in liver and skin. Dexamethasone was also administered by intraperitoneal injection. Topical application or intraperitoneal injection of dexamethasone increased cutaneous CYP2E1 (8 and 4-fold respectively) whereas the hepatic level of this isoform showed a slight decrease and hepatic CYP3A expression was increased (3-fold). Cutaneous CYP2E1 was increased (3-fold) after topical treatment by salicylic acid. This compound had no effect on hepatic CYP3A and CYP2E1 expression. Cutaneous CYP3A (protein and mRNA) was not detectable in all groups (control or treated animals). Dexamethasone and salicylic acid increased cutaneous CYP2E1 mRNA level (2.5 and 1.4-fold respectively). In conclusion, dexamethasone and salicylic acid induced cutaneous CYP2E1 protein and mRNA level. Cutaneous CYP2E1 induction by dexamethasone is a tissue-specific process. PMID:9207195

  7. Involvement of CYP 2E1 enzyme in ovotoxicity caused by 4-vinylcyclohexene and its metabolites

    SciTech Connect

    Rajapaksa, Kathila S.; Cannady, Ellen A.; Sipes, I. Glenn; Hoyer, Patricia B. . E-mail: hoyer@u.arizona.edu

    2007-06-01

    4-Vinylcyclohexene (VCH) is bioactivated by hepatic CYP 2A and 2B to a monoepoxide (VCM) and subsequently to an ovotoxic diepoxide metabolite (VCD). Studies suggest that the ovary can directly bioactivate VCH via CYP 2E1. The current study was designed to evaluate the role of ovarian CYP 2E1 in VCM-induced ovotoxicity. Postnatal day 4 B6C3F{sub 1} and CYP 2E1 wild-type (+/+) and null (-/-) mouse ovaries were cultured (15 days) with VCD (30 {mu}M), 1,2-VCM (125-1000 {mu}M), or vehicle. Twenty-eight days female CYP 2E1 +/+ and -/- mice were dosed daily (15 days; ip) with VCH, 1,2-VCM, VCD or vehicle. Following culture or in vivo dosing, ovaries were histologically evaluated. In culture, VCD decreased (p < 0.05) primordial and primary follicles in ovaries from all three groups of mice. 1,2-VCM decreased (p < 0.05) primordial follicles in B6C3F{sub 1} and CYP 2E1 +/+ ovaries, but not in CYP 2E1 -/- ovaries in culture. 1,2-VCM did not affect primary follicles in any group of mouse ovaries. Conversely, following in vivo dosing, primordial and primary follicles were reduced (p < 0.05) by VCD and VCM in CYP2E1 +/+ and -/-, and by VCH in +/+ mice. The data demonstrate that, whereas in vitro ovarian bioactivation of VCM requires CYP 2E1 enzyme, in vivo CYP 2E1 plays a minimal role. Thus, the findings support that hepatic metabolism dominates the contribution made by the ovary in bioactivation of VCM to its ovotoxic metabolite, VCD. This study also demonstrates the use of a novel ovarian culture system to evaluate ovary-specific metabolism of xenobiotics.

  8. Involvement of CYP 2E1 enzyme in ovotoxicity caused by 4-vinylcyclohexene and its metabolites.

    PubMed

    Rajapaksa, Kathila S; Cannady, Ellen A; Sipes, I Glenn; Hoyer, Patricia B

    2007-06-01

    4-Vinylcyclohexene (VCH) is bioactivated by hepatic CYP 2A and 2B to a monoepoxide (VCM) and subsequently to an ovotoxic diepoxide metabolite (VCD). Studies suggest that the ovary can directly bioactivate VCH via CYP 2E1. The current study was designed to evaluate the role of ovarian CYP 2E1 in VCM-induced ovotoxicity. Postnatal day 4 B6C3F(1) and CYP 2E1 wild-type (+/+) and null (-/-) mouse ovaries were cultured (15 days) with VCD (30 microM), 1,2-VCM (125-1000 microM), or vehicle. Twenty-eight days female CYP 2E1 +/+ and -/- mice were dosed daily (15 days; ip) with VCH, 1,2-VCM, VCD or vehicle. Following culture or in vivo dosing, ovaries were histologically evaluated. In culture, VCD decreased (p<0.05) primordial and primary follicles in ovaries from all three groups of mice. 1,2-VCM decreased (p<0.05) primordial follicles in B6C3F(1) and CYP 2E1 +/+ ovaries, but not in CYP 2E1 -/- ovaries in culture. 1,2-VCM did not affect primary follicles in any group of mouse ovaries. Conversely, following in vivo dosing, primordial and primary follicles were reduced (p<0.05) by VCD and VCM in CYP2E1 +/+ and -/-, and by VCH in +/+ mice. The data demonstrate that, whereas in vitro ovarian bioactivation of VCM requires CYP 2E1 enzyme, in vivo CYP 2E1 plays a minimal role. Thus, the findings support that hepatic metabolism dominates the contribution made by the ovary in bioactivation of VCM to its ovotoxic metabolite, VCD. This study also demonstrates the use of a novel ovarian culture system to evaluate ovary-specific metabolism of xenobiotics. PMID:17462685

  9. Strength in numbers.

    PubMed

    Lakkis, S

    1999-03-01

    A case study showing the attitudes and actions reinforcing discrimination against women's rights in Lebanon is presented. The study illustrates the way in which the public¿s views and the interests of families and local dignitaries can manipulate opinions. Organizations aimed at protecting women's rights have found strength in working together. The Lebanese League, an organization comprised of 17 women's and human rights associations, has established a center and a telephone hotline to encourage abused women to disclose and discuss their situation. The center provides support in the form of legal, psychological and medical assistance. Another organization working with the Lebanese League towards the same vision is the Lebanese Physically Handicapped Union. Efforts to lobby around a Lebanese legislation discriminating against women so far had no success, but all organizations involved are aware of the need to work through a range of interventions, and to take a long-term view, before they can count on any success. PMID:12295032

  10. QM02 Strength Measurement

    SciTech Connect

    Welch, J; Wu, J.; ,

    2010-11-24

    In late April, Paul Emma reported that his orbit fitting program could find a reasonably good fit only if the strength of QM02 was changed from design value of -5.83 kG to -6.25 kG - a strength change of 7.3%. In late May, we made a focal length measurement of QM02 by turning off all focusing optics between YC07 and BPMS1 (in the spectrometer line) except for QM02 and adjusted the strength of QM02 so that vertical kicks by YC07 did not produce any displacements at BPMS1 (see Figure 1). The result was quoted in the LCLS elog was that QM02 appeared to 6% too weak, and approximately agreed with Paul's observation. The analysis used for the entry in the log book was based on the thin lens approximation and used the following numbers: Distance YC07 to QM02 - 5.128 m; Distance QM02 to BPMS1 - 1.778 m; and Energy - 135 MeV. These distances were computed from the X,Z coordinates given the on the large plot of the Injector on the wall of the control room. On review of the MAD output file coordinates, it seems that the distance used for QM02 to BPMS1 is not 1.778 m. The correct value is Distance, center of QM02 to BPMS1 - 1.845 m. There may be a typo on the wall chart values for the coordinates of BPMS1, or perhaps there was a misinterpretation of edge versus center of QM02. In any case, the effect of this change is that the thin lens estimate changes from 6% too weak to 9% too weak. At John Galayda's suggestion, we looked into the thin lens versus thick lens approximation. A Mathematica program was written to solve for the K value of the QM02, in the thick lens approximation, that provides point to point focusing from YC07 to BPMS1, and to compare this number with the value obtained using the thin lens approximation. The length of QM02 used in the thick lens calculation is the effective length determined by magnetic measurements of 0.108 m. The result of the Mathematica calculation is that the thin lens approximation predicts less magnet strength is required to produce the same focusing by about 1.3%. When both the distance correction and the thick lens approximation are taken into account, the result is: K{sub thick} - K{sub mm}/K{sub thick} = -7.6% where K{sub mm} is the value obtained from magnetic measurements and K{sub thick} is the value of K obtained from the focal length measurement in the thick lens approximation. That is, QM02 acts weaker than it was measured by magnetic measurements by 7.6%. This is remarkably close to Paul's original estimate. The unexpected weakness could in principle be due to several things: shorted turns, a current calibration error, magnetic measurement error; it could even be due to the presence of an gradient from QM01, which is of opposite sign and right next to QM02, despite it being set to zero current (although it was not DAC-zeroed). Plans have been implemented to remove and replace QM02 at the earliest ROD.

  11. [The strength of fragile].

    PubMed

    Lagrée, J

    2015-02-01

    Chronic disorder not only modifies the patient's health status; it affects the body relationship and its potential; it decreases its power to act, changes its relation to time, to its relatives, in shortcut it affects all dimensions of the relationship with oneself, with others, and the world. However, if we assume that a patient cannot be reduced to his/her illness but must always be treated with the respect of a person, we also understand how chronic and debilitating disease affects both freedom (or autonomy) and perfectibility (or creativity). By analyzing the different forms of freedom of the patient, this paper intends to rehabilitate the fragile own strength which is fortitude. It manifests itself in attitudes and virtues such as gentleness, patience, attention to the present, hope, power of thought. It also intends to show that it is supporting fragile and allowing them to develop their own way of acting and living as humanity became intelligent and creative. PMID:24721121

  12. High strength ferritic alloy

    DOEpatents

    Hagel, William C.; Smidt, Frederick A.; Korenko, Michael K.

    1977-01-01

    A high-strength ferritic alloy useful for fast reactor duct and cladding applications where an iron base contains from about 9% to about 13% by weight chromium, from about 4% to about 8% by weight molybdenum, from about 0.2% to about 0.8% by weight niobium, from about 0.1% to about 0.3% by weight vanadium, from about 0.2% to about 0.8% by weight silicon, from about 0.2% to about 0.8% by weight manganese, a maximum of about 0.05% by weight nitrogen, a maximum of about 0.02% by weight sulfur, a maximum of about 0.02% by weight phosphorous, and from about 0.04% to about 0.12% by weight carbon.

  13. Production of high strength concrete

    SciTech Connect

    Peterman, M.B.; Carrasquillo, R.L.

    1986-01-01

    The criteria for selection of concrete materials and their proportions to producer uniform, economical, high strength concrete are presented in this book. The recommendations provided are based on a study of the interactions among components of plain concrete and mix proportions, and of their contribution to the compressive strength of high strength concrete. These recommendations will serve as guidelines to practicing engineers, in the selection of materials and their proportions for the production of high strength concrete. Increasing demands for improved efficiency and reduced construction costs have resulted in engineers beginning to design large structures using higher strength concrete at higher stress levels. There are definite advantages, both technical and economical, in using high strength concrete. For example, for a given cross section, prestresses concrete bridge girders can carry greater service loads across longer spans if made using high strength concrete. In addition, cost comparisons have shown that the savings obtained are significantly greater than the added cost of the higher quality concrete.

  14. DprE1 Is a Vulnerable Tuberculosis Drug Target Due to Its Cell Wall Localization.

    PubMed

    Brecik, Miroslav; Centárová, Ivana; Mukherjee, Raju; Kolly, Gaëlle S; Huszár, Stanislav; Bobovská, Adela; Kilacsková, Emöke; Mokošová, Veronika; Svetlíková, Zuzana; Šarkan, Michal; Neres, João; Korduláková, Jana; Cole, Stewart T; Mikušová, Katarína

    2015-07-17

    The flavo-enzyme DprE1 catalyzes a key epimerization step in the decaprenyl-phosphoryl d-arabinose (DPA) pathway, which is essential for mycobacterial cell wall biogenesis and targeted by several new tuberculosis drug candidates. Here, using differential radiolabeling with DPA precursors and high-resolution fluorescence microscopy, we disclose the unexpected extracytoplasmic localization of DprE1 and periplasmic synthesis of DPA. Collectively, this explains the vulnerability of DprE1 and the remarkable potency of the best inhibitors. PMID:25906160

  15. Suppression of mutations in two Saccharomyces cerevisiae genes by the adenovirus E1A protein.

    PubMed

    Zieler, H A; Walberg, M; Berg, P

    1995-06-01

    The protein products of the adenoviral E1A gene are implicated in a variety of transcriptional and cell cycle events, involving interactions with several proteins present in human cells, including parts of the transcriptional machinery and negative regulators of cell division such as the Rb gene product and p107. To determine if there are functional homologs of E1A in Saccharomyces cerevisiae, we have developed a genetic screen for mutants that depend on E1A for growth. The screen is based on a colony color sectoring assay which allows the identification of mutants dependent on the maintenance and expression of an E1A-containing plasmid. Using this screen, we have isolated five mutants that depend on expression of the 12S or 13S cDNA of E1A for growth. A plasmid shuffle assay confirms that the plasmid-dependent phenotype is due to the presence of either the 12S or the 13S E1A cDNA and that both forms of E1A rescue growth of all mutants equally well. The five mutants fall into two classes that were named web1 and web2 (for "wants E1A badly"). Plasmid shuffle assays with mutant forms of E1A show that conserved region 1 (CR1) is required for rescue of the growth of the web1 and web2 E1A-dependent yeast mutants, while the N-terminal 22 amino acids are only partially required; conserved region 2 (CR2) and the C terminus are dispensable. The phenotypes of mutants in both the web1 and the web2 groups are due to a single gene defect, and the yeast genes that fully complement the mutant phenotypes of both groups were cloned. The WEB1 gene sequence encodes a 1,273-amino-acid protein that is identical to SEC31, a protein involved in the budding of transport vesicles from the endoplasmic reticulum. The WEB2 gene encodes a 1,522-amino-acid protein with homology to nucleic acid-dependent ATPases. Deletion of either WEB1 or WEB2 is lethal. Expression of E1A is not able to rescue the lethality of either the web1 or the web2 null allele, implying allele-specific mutations that lead to E1A dependence. PMID:7760818

  16. 26 CFR 48.6416(e)-1 - Refund to exporter or shipper.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) MISCELLANEOUS EXCISE TAXES MANUFACTURERS AND RETAILERS EXCISE TAXES Refunds and Other Administrative Provisions of Special Application to Retailers and Manufacturers Taxes § 48.6416(e)-1 Refund to exporter...

  17. Hepatitis C Virus Envelope Glycoprotein E1 Forms Trimers at the Surface of the Virion

    PubMed Central

    Falson, Pierre; Bartosch, Birke; Alsaleh, Khaled; Tews, Birke Andrea; Loquet, Antoine; Ciczora, Yann; Riva, Laura; Montigny, Cédric; Montpellier, Claire; Duverlie, Gilles; Pécheur, Eve-Isabelle; le Maire, Marc; Cosset, François-Loïc

    2015-01-01

    ABSTRACT In hepatitis C virus (HCV)-infected cells, the envelope glycoproteins E1 and E2 assemble as a heterodimer. To investigate potential changes in the oligomerization of virion-associated envelope proteins, we performed SDS-PAGE under reducing conditions but without thermal denaturation. This revealed the presence of SDS-resistant trimers of E1 in the context of cell-cultured HCV (HCVcc) as well as in the context of HCV pseudoparticles (HCVpp). The formation of E1 trimers was found to depend on the coexpression of E2. To further understand the origin of E1 trimer formation, we coexpressed in bacteria the transmembrane (TM) domains of E1 (TME1) and E2 (TME2) fused to reporter proteins and analyzed the fusion proteins by SDS-PAGE and Western blotting. As expected for strongly interacting TM domains, TME1–TME2 heterodimers resistant to SDS were observed. These analyses also revealed homodimers and homotrimers of TME1, indicating that such complexes are stable species. The N-terminal segment of TME1 exhibits a highly conserved GxxxG sequence, a motif that is well documented to be involved in intramembrane protein-protein interactions. Single or double mutations of the glycine residues (Gly354 and Gly358) in this motif markedly decreased or abrogated the formation of TME1 homotrimers in bacteria, as well as homotrimers of E1 in both HCVpp and HCVcc systems. A concomitant loss of infectivity was observed, indicating that the trimeric form of E1 is essential for virus infectivity. Taken together, these results indicate that E1E2 heterodimers form trimers on HCV particles, and they support the hypothesis that E1 could be a fusion protein. IMPORTANCE HCV glycoproteins E1 and E2 play an essential role in virus entry into liver cells as well as in virion morphogenesis. In infected cells, these two proteins form a complex in which E2 interacts with cellular receptors, whereas the function of E1 remains poorly understood. However, recent structural data suggest that E1 could be the protein responsible for the process of fusion between viral and cellular membranes. Here we investigated the oligomeric state of HCV envelope glycoproteins. We demonstrate that E1 forms functional trimers after virion assembly and that in addition to the requirement for E2, a determinant for this oligomerization is present in a conserved GxxxG motif located within the E1 transmembrane domain. Taken together, these results indicate that a rearrangement of E1E2 heterodimer complexes likely occurs during the assembly of HCV particles to yield a trimeric form of the E1E2 heterodimer. Gaining structural information on this trimer will be helpful for the design of an anti-HCV vaccine. PMID:26246575

  18. Ber e 1 protein: the versatile major allergen from Brazil nut seeds.

    PubMed

    Alcocer, Marcos; Rundqvist, Louise; Larsson, Göran

    2012-04-01

    Due mainly to its extremely high content of sulphur amino acids, Ber e 1 protein, the major allergen from Brazil nut, has attracted much scientific and press attention. Ber e 1 was the main target protein in early biotechnology transgenic work, in early processing studies of plant storage proteins, in plant vacuolar targeting studies and as the main protein in early nutritional supplementation experiments. Ber e 1 was also one of the first food allergens to be unintentionally transferred from one plant to another and was involved in the first reported case of systemic allergic reaction caused by a food allergen transferred in semen. In this review, many of the Ber e 1 unique biotechnological and structural functions are discussed with a particular emphasis on its use as model protein for studies of intrinsic allergenicity of food proteins. PMID:22187079

  19. Effects of orbital and spin current interference in E1 and M2 nuclear excitations

    SciTech Connect

    Goncharova, N. G.

    2015-12-15

    The interference of contributions from the orbital and spin currents to the E1 and M2 resonances is investigated. The results of the current interference analysis within the shell model are compared with the experimental data.

  20. Impact strength and indentation hardness of high-strength ceramics

    SciTech Connect

    Grady, D.E.

    1993-08-01

    Hugoniot elastic limit and indentation hardness data are provided for selected high-strength ceramics AlN, Al{sub 2}O{sub 3}, B{sub 4}C, SiC, TiB, Zr{sub 2}. Degree of correlation of the strength data by the two test methods is examined. Influence of reformation mechanism on strength measurement, including rate sensitivity, size scale and confining pressure, is discussed.

  1. Natural history of the E1-like superfamily: implication for adenylation, sulfur transfer and ubiquitin conjugation

    PubMed Central

    Burroughs, A. Maxwell; Iyer, Lakshminarayan M.; Aravind, L.

    2009-01-01

    The E1-like superfamily is central to ubiquitin (Ub) conjugation, biosynthesis of cysteine, thiamine and MoCo and several secondary metabolites. Yet, its functional diversity and evolutionary history is not well-understood. We develop a natural classification of this superfamily and use it to decipher the major adaptive trends occurring in the evolution of the E1-like superfamily. Within the Rossmann fold, E1-like proteins are closest to NAD(P)/FAD-dependent dehydrogenases and S-AdoMet-dependent methyltransferases. Hence, their phosphotransfer activity is an independent catalytic “invention” with respect to such activities seen in other Rossmannoid folds. Sequence and structure analysis reveals a striking diversity of residues and structures involved in adenylation, sulfotransfer and substrate-binding between different E1-like families, allowing us to predict previously uncharacterized functional adaptations. E1-like proteins are fused to several previously undetected domains, such as a predicted sulfur transfer domain containing a novel superfamily of the TATA-binding protein fold, different types of catalytic domains, a novel winged helix-turn-helix domain and potential adaptor domains related to Ub conjugation. Based on these fusions we develop a generalized model for the linking of E1 catalyzed adenylation/thiolation with further down-stream reactions. This is likely to involve a dynamic interplay between the E1 active sites and diverse fused C-terminal domains. We also predict participation of E1-like domains in previously uncharacterized bacterial secondary metabolism pathways, new cysteine biosynthesis systems, such as those associated with archaeal O-phosphoseryl tRNA, metal-sulfur cluster assembly (e.g. in nitrogen fixation) and Ub-conjugation. Evolutionary reconstructions suggest that the last universal common ancestor (LUCA) contained a single E1-like domain possessing both phosphotransfer and thiolating activities and participating in multiple sulfotransfer reactions. The E1-like superfamily subsequently expanded to include 26 families clustering into three major radiations. These are broadly involved in ubiquitin activation, cofactor and cysteine biosynthesis, and biosynthesis of secondary metabolites. In light of this we present evidence that in eukaryotes other E1-like enzymes, such as Urm1, were independently recruited for Ubl conjugation, probably functioning without conventional E2-like enzymes. PMID:19089947

  2. Resolvin E1 Improves Tear Production and Decreases Inflammation in a Dry Eye Mouse Model

    PubMed Central

    Li, Na; He, Jiucheng; Schwartz, Carl Eric; Gjorstrup, Per

    2010-01-01

    Abstract Purpose Dry eye (DE) is a common ocular surface disease, particularly among women and the elderly, with chronic symptoms of eye irritation and, in severe cases, blurred vision. Several studies have shown that there is an inflammatory component in DE, although the pathogenesis is not thoroughly understood. Resolvin E1 (RvE1; RX-10001) is an endogenous mediator derived from the omega-3 polyunsaturated fatty acid eicosapentaenoic acid and is involved in inflammation resolution and tissue protection. Here we investigated the role of RvE1 in a DE mouse model. Methods Thirteen- to 14-week-old female BALB/C mice were exposed to desiccating conditions. One week after DE exposure, animals were treated topically with drug or vehicle 4 times per day for an additional week. Controls were nontreated animals placed in a normal environment. Schirmer's test was performed before treatment initiation and at days 2 and 4 after treatment. Density of corneal epithelial cells was analyzed in vivo using the Rostock Cornea Module of the Heidelberg Retina Tomograph (HRT-II). Corneas were processed using Western blot analysis and immunofluorescence examination. Results Schirmer's test showed a significant decrease in tear production in DE compared with controls. There was no change at 2 and 4 days after treatment with the vehicle, but a significant increase was observed at 2 and 4 days in the RvE1-treated group. The density of the superficial epithelial cells showed a significant decrease after DE compared with controls, which increased after 7 days of RvE1 treatment. Western blot analysis showed that α-smooth muscle actin and cyclooxygenase-2 (COX-2) expression were strongly upregulated after DE and decreased after 7 days of RvE1 treatment. Immunofluorescence confirmed strong positive staining of α-smooth muscle actin and COX-2 in stroma and/or in epithelia after DE, which decreased with RvE1 treatment. The percentage of infiltrating CD4+ T cells and CD11b+ cells decreased after RvE1 treatment when compared with DE. Conclusion RvE1 promotes tear production, corneal epithelial integrity, and a decrease in inflammatory inducible COX-2. In the stroma, RvE1 inhibits keratocyte transformation to myofibroblasts and lowers the number of monocytes/macrophages in this DE mouse model. These results suggest that RvE1 and similar resolvin analogs have therapeutic potential in the treatment of DE. PMID:20874497

  3. Ethanol Induction of CYP2A5: Permissive Role for CYP2E1

    PubMed Central

    Lu, Yongke; Zhuge, Jian; Wu, Defeng

    2011-01-01

    CYP2A5 metabolizes xenobiotics and activates hepatocarcinogens, and induction occurs in response to hepatic damage and cellular stress. We evaluated whether ethanol can elevate CYP2A5 and whether CYP2E1 plays a role in the ethanol induction of CYP2A5. Wild-type (WT), CYP2E1 knockout (KO), and CYP2E1 knockin (KI) mice were fed ethanol for 3 weeks. Ethanol increased CYP2E1 and CYP2A5 protein and activity in WT mice but not in the KO mice. Induction of CYP2A5 (and CYP2E1) was restored in the KI mice. Ethanol induction of CYP2A5 occurred only after CYP2E1 was first induced. Immunohistochemical staining revealed that CYP2E1 and CYP2A5 colocalize to the same zones in the liver. Ethanol also elevated CYP2A5 mRNA levels in WT and KI mice but not in KO mice. Induction of CYP2A5 by cadmium was partially decreased in KO mice compared with WT or KI mice. Ethanol elevated CYP2A4 mRNA levels in all mice although the extent of induction was lowest in the KO mice. In summary, ethanol elevated mouse hepatic CYP2A5 levels, which may be of toxicological significance because CYP2A5 metabolizes nicotine and other drugs and activates hepatocarcinogens. Induction of CYP2A5 by ethanol is potentiated by the induction of CYP2E1. We speculate that ethanol induction of CYP2E1 followed by increases in reactive oxygen species and activation of Nrf2 are important steps in the mechanism by which ethanol induces CYP2A5. The possibility that induction of CYP2E1 is permissive for the induction of CYP2A5 may reflect a new contribution by CYP2E1 to the actions of ethanol. PMID:21051534

  4. Impact of Resolvin E1 on Murine Neutrophil Phagocytosis in Type 2 Diabetes

    PubMed Central

    Herrera, Bruno S.; Hasturk, Hatice; Kantarci, Alpdogan; Freire, Marcelo O.; Nguyen, Olivia; Kansal, Shevali

    2014-01-01

    Diabetic complications involve inflammation-mediated microvascular and macrovascular damage, disruption of lipid metabolism, glycosylation of proteins, and abnormalities of neutrophil-mediated events. Resolution of inflamed tissues to health and homeostasis is an active process mediated by endogenous lipid agonists, including lipoxins and resolvins. This proresolution system appears to be compromised in type 2 diabetes (T2D). The goal of this study was to investigate unresolved inflammation in T2D. Wild-type (WT) and genetically engineered mice, including T2D mice (db/db), transgenic mice overexpressing the human resolvin E1 (RvE1) receptor (ERV1), and a newly bred strain of db/ERV1 mice, were used to determine the impact of RvE1 on the phagocytosis of Porphyromonas gingivalis in T2D. Neutrophils were isolated and incubated with fluorescein isothiocyanate-labeled P. gingivalis, and phagocytosis was measured in a fluorochrome-based assay by flow cytometry. Mitogen-activated protein kinase (MAPK) (p42 and p44) and Akt (Thr308 and Ser473) phosphorylation was analyzed by Western blotting. The mouse dorsal air pouch model was used to evaluate the in vivo impact of RvE1. Results revealed that RvE1 increased the neutrophil phagocytosis of P. gingivalis in WT animals but had no impact in db/db animals. In ERV1-transgenic and ERV1-transgenic diabetic mice, phagocytosis was significantly increased. RvE1 decreased Akt and MAPK phosphorylation in the transgenic animals. In vivo dorsal air pouch studies revealed that RvE1 decreases neutrophil influx into the pouch and increases neutrophil phagocytosis of P. gingivalis in the transgenic animals; cutaneous fat deposition was reduced, as was macrophage infiltration. The results suggest that RvE1 rescues impaired neutrophil phagocytosis in obese T2D mice overexpressing ERV1. PMID:25486994

  5. Structure-function analysis of hepatitis C virus envelope glycoproteins E1 and E2.

    PubMed

    Nayak, Aparajita; Pattabiraman, Nagarajan; Fadra, Numrah; Goldman, Radoslav; Kosakovsky Pond, Sergei L; Mazumder, Raja

    2015-01-01

    Hepatitis C virus (HCV) is the leading cause of chronic liver disease in humans. The envelope proteins of HCV are potential candidates for vaccine development. The absence of three-dimensional (3D) structures for the functional domain of HCV envelope proteins [E1.E2] monomer complex has hindered overall understanding of the virus infection, and also structure-based drug design initiatives. In this study, we report a 3D model containing both E1 and E2 proteins of HCV using the recently published structure of the core domain of HCV E2 and the functional part of E1, and investigate immunogenic implications of the model. HCV [E1.E2] molecule is modeled by using aa205-319 of E1 to aa421-716 of E2. Published experimental data were used to further refine the [E1.E2] model. Based on the model, we predict 77 exposed residues and several antigenic sites within the [E1.E2] that could serve as vaccine epitopes. This study identifies eight peptides which have antigenic propensity and have two or more sequentially exposed amino acids and 12 singular sites are under negative selection pressure that can serve as vaccine or therapeutic targets. Our special interest is 285FLVGQLFTFSPRRHW299 which has five negatively selected sites (L286, V287, G288, T292, and G303) with three of them sequential and four amino acids exposed (F285, L286, T292, and R296). This peptide in the E1 protein maps to dengue envelope vaccine target identified previously by our group. Our model provides for the first time an overall view of both the HCV envelope proteins thereby allowing researchers explore structure-based drug design approaches. PMID:25245635

  6. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... Equivalent Methods for PM2.5 or PM10â2.5 Pt. 53, Subpt. E, Fig. E-1 Figure E-1 to Subpart E of Part 53... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who... 40 Protection of Environment 5 2011-07-01 2011-07-01 false Designation Testing Checklist E...

  7. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... Equivalent Methods for PM2.5 or PM10â2.5 Pt. 53, Subpt. E, Fig. E-1 Figure E-1 to Subpart E of Part 53... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who...-7.4.10) Application Specification Tests Field Precision (§ 53.58) (L-5.1) Meets all Appendix...

  8. [CYP2E1 gene polymorphism and ovarian cancer risk in the Yakut population].

    PubMed

    Khrunin, A V; Ivanova, F G; Moiseev, A A; Gorbunova, V A; Limborskaia, S A

    2011-12-01

    The CYP2E1 gene polymorphism has been studied in Yakut women with ovarian cancer and without cancer. The two groups have been found to substantially differ in the frequency of the CYP2E1* 1D allele (with a 96-bp insertion in the promoter region of the gene): it is more frequent in healthy women (16.3 versus 7.4%, P = 0.007). PMID:22384697

  9. Mammalian PC-12 cell genetically engineered for human cytochrome P450 2E1 expression.

    PubMed

    Mapoles, J; Berthou, F; Alexander, A; Simon, F; Ménez, J F

    1993-06-15

    The stable expression of the human cytochrome CYP2E1 (P450 alcohol) was performed in the mammalian cell line PC-12. This cell line expressed cytochrome b5 (58 +/- 12 pmol/mg microsomal protein vs 528 +/- 80 pmol/mg in microsomal human liver) and a high level of NADPH: cytochrome P450 reductase (140 +/- 20 nmol.min-1.mg microsomal protein-1 vs 68 +/- 48 nmol.min-1.mg-1 in microsomal human liver). An expression plasmid was constructed using the cDNA for the human CYP2E1 mRNA and the Rous sarcoma virus (RSV) promoter. This plasmid was co-transfected with the plasmid RSVneo into PC-12 cells. Clones were selected for resistance to the neomycin analog, G418, and then screened for expression of the CYP2E1 isozyme by testing for 6-hydroxylation of chlorzoxazone, a specific substrate for CYP2E1. Expression of CYP2E1 was confirmed in one clone, DB-7, by Western blot analysis and by measurement of monooxygenase activities which were not detectable in PC-12 cells. Chlorzoxazone 6-hydroxylation, n-butanol oxidation and dimethylnitrosamine N-demethylation were localized in microsomes (62, 60 and 63 pmol.min-1.mg microsomal protein-1, respectively) and were inhibited by carbon monoxide and diethyldithiocarbamate, both inhibitors of P450 enzymes. Although the level of the enzyme activities was about a tenth of that measured in human liver microsomes, CYP2E1 expressed in DB-7 cells has catalytic competence similar to human liver CYP2E1. DB-7 cells metabolized acetaminophen and this metabolic activation was shown to be toxic to these cells by release of lactate dehydrogenase. Construction of recombinant cell lines expressing CYP2E1 provides a useful tool for studying the catalytic properties of this enzyme and the consequent cytotoxic effects of substrates metabolized by this enzyme. PMID:8391436

  10. 11 CFR 300.60 - Scope (2 U.S.C. 441i(e)(1)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) REGULATIONS NON-FEDERAL FUNDS Federal Candidates and Officeholders § 300.60 Scope (2 U.S.C. 441i(e)(1)). This subpart applies to: (a) Federal candidates; (b) Individuals holding Federal office (see 11 CFR 300.2(o... 11 Federal Elections 1 2010-01-01 2010-01-01 false Scope (2 U.S.C. 441i(e)(1)). 300.60 Section...

  11. 11 CFR 300.60 - Scope (2 U.S.C. 441i(e)(1)).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...) REGULATIONS NON-FEDERAL FUNDS Federal Candidates and Officeholders § 300.60 Scope (2 U.S.C. 441i(e)(1)). This subpart applies to: (a) Federal candidates; (b) Individuals holding Federal office (see 11 CFR 300.2(o... 11 Federal Elections 1 2011-01-01 2011-01-01 false Scope (2 U.S.C. 441i(e)(1)). 300.60 Section...

  12. Fault Roughness Records Strength

    NASA Astrophysics Data System (ADS)

    Brodsky, E. E.; Candela, T.; Kirkpatrick, J. D.

    2014-12-01

    Fault roughness is commonly ~0.1-1% at the outcrop exposure scale. More mature faults are smoother than less mature ones, but the overall range of roughness is surprisingly limited which suggests dynamic control. In addition, the power spectra of many exposed fault surfaces follow a single power law over scales from millimeters to 10's of meters. This is another surprising observation as distinct structures such as slickenlines and mullions are clearly visible on the same surfaces at well-defined scales. We can reconcile both observations by suggesting that the roughness of fault surfaces is controlled by the maximum strain that can be supported elastically in the wallrock. If the fault surface topography requires more than 0.1-1% strain, it fails. Invoking wallrock strength explains two additional observations on the Corona Heights fault for which we have extensive roughness data. Firstly, the surface is isotropic below a scale of 30 microns and has grooves at larger scales. Samples from at least three other faults (Dixie Valley, Mount St. Helens and San Andreas) also are isotropic at scales below 10's of microns. If grooves can only persist when the walls of the grooves have a sufficiently low slope to maintain the shape, this scale of isotropy can be predicted based on the measured slip perpendicular roughness data. The observed 30 micron scale at Corona Heights is consistent with an elastic strain of 0.01 estimated from the observed slip perpendicular roughness with a Hurst exponent of 0.8. The second observation at Corona Heights is that slickenlines are not deflected around meter-scale mullions. Yielding of these mullions at centimeter to meter scale is predicted from the slip parallel roughness as measured here. The success of the strain criterion for Corona Heights supports it as the appropriate control on fault roughness. Micromechanically, the criterion implies that failure of the fault surface is a continual process during slip. Macroscopically, the fundamental nature of the control means that 0.1 to 1% roughness should be ubiquitous on faults and can generally be used for simulating ground motion. An important caveat is that the scale-dependence of strength may result in a difference in the yield criterion at large-scales. The commonly observed values of the Hurst exponent below 1 may capture this scale-dependence.

  13. Specific uptake of radioiodinated fragment E1 by venous thrombi in pigs.

    PubMed Central

    Knight, L C; Olexa, S A; Malmud, L S; Budzynski, A Z

    1983-01-01

    Fragment E1, a product of plasmic digestion of cross-linked fibrin, binds specifically in vitro to polymerized fibrin but not to fibrinogen. Purified human Fragment E1 was radiolabeled with 125I or 131I by the Iodogen technique. The uptake of radioiodinated Fragment E1 in vitro into forming or preformed clots was demonstrated. Animal biodistribution studies of radioiodinated Fragment E1 showed its rapid removal from the circulation; radioactive catabolites did not reside long in any organ and were excreted in the urine. The uptake in vivo was evaluated in pigs with preexisting venous thrombi of various ages from 1 h up to 5 d at the time of intravenous systemic injection of the tracer. Radioiodinated fibrinogen was also injected into the same animals to compare the uptake of the two tracers. Thrombus-to-blood ratios for Fragment E1 averaged 43:1 (range 10-108) and 29:1 (range 8-107) in thrombi 1-6 h and 1-5 d old, respectively. In contrast, mean thrombus-to-blood ratios for fibrinogen were, in the same time intervals, 26:1 (range 17-41) and 2:1 (range 0.5-3.9), respectively. It is concluded that radioiodinated Fragment E1 is a specific marker of thrombi in vivo: its uptake by fresh thrombi is better than that of labeled fibrinogen and, in contrast to radioiodinated fibrinogen, this fragment is incorporated into old thrombi as well. PMID:6643683

  14. In vitro characterization of the NAD(+) synthetase NadE1 from Herbaspirillum seropedicae.

    PubMed

    Laskoski, Kerly; Santos, Adrian R S; Bonatto, Ana C; Pedrosa, Fábio O; Souza, Emanuel M; Huergo, Luciano F

    2016-05-01

    Nicotinamide adenine dinucleotide synthetase enzyme (NadE) catalyzes the amination of nicotinic acid adenine dinucleotide (NaAD) to form NAD(+). This reaction represents the last step in the majority of the NAD(+) biosynthetic routes described to date. NadE enzymes typically use either glutamine or ammonium as amine nitrogen donor, and the reaction is energetically driven by ATP hydrolysis. Given the key role of NAD(+) in bacterial metabolism, NadE has attracted considerable interest as a potential target for the development of novel antibiotics. The plant-associative nitrogen-fixing bacteria Herbaspirillum seropedicae encodes two putative NadE, namely nadE1 and nadE2. The nadE1 gene is linked to glnB encoding the signal transduction protein GlnB. Here we report the purification and in vitro characterization of H. seropedicae NadE1. Gel filtration chromatography analysis suggests that NadE1 is an octamer. The NadE1 activity was assayed in vitro, and the Michaelis-Menten constants for substrates NaAD, ATP, glutamine and ammonium were determined. Enzyme kinetic and in vitro substrate competition assays indicate that H. seropedicae NadE1 uses glutamine as a preferential nitrogen donor. PMID:26802007

  15. ATF4 deficiency protects hepatocytes from oxidative stress via inhibiting CYP2E1 expression

    PubMed Central

    Wang, Chunxia; Li, Houkai; Meng, Qingshu; Du, Ying; Xiao, Fei; Zhang, Qian; Yu, Junjie; Li, Kai; Chen, Shanghai; Huang, Zhiying; Liu, Bin; Guo, Feifan

    2014-01-01

    Activating transcription factor (ATF) 4 is involved in the regulation of oxidative stress in fibroblasts and neurons. The role of ATF4 in hepatocytes, however, is unknown. The aim of this study was to investigate the role of ATF4 in hepatocytes in oxidative stress under a high-fat diet (HFD). Here, we showed that palmitate-stimulated reactive oxygen species (ROS) production and triglyceride (TG) accumulation is blocked by ATF4 deficiency in primary hepatocytes. Consistently, HFD-induced oxidative stress, TG accumulation and expression of cytochrome P450, family 2, subfamily, polypeptide 1 (CYP2E1) are also blocked by knocking down ATF4 expression in the mouse liver. This suggests that ATF4 might regulate oxidative stress viaCYP2E1 under an HFD. In addition, we observed that expression of CYP2E1 is indirectly regulated by ATF4 in a cAMP-responsive element binding protein (CREB)-dependent manner, which can directly activate the CYP2E1 promoter activity. Notably, ATF4-stimulated ROS production is inhibited in vivo by treatment with diallyl sulphide, a selective CYP2E1 inhibitor. Finally, we showed that ATF4 expression in the liver is responsible for the protective effects against HFD-induced CYP2E1 expression, oxidative stress, and TG accumulation. Taken together, these observations suggest that ATF4 is a novel regulator of oxidative stress as well as accumulation of TG in response to HFD. PMID:24373582

  16. Cyclin/CDK regulates the nucleocytoplasmic localization of the human papillomavirus E1 DNA helicase.

    PubMed

    Deng, Wentao; Lin, Biing Yuan; Jin, Ge; Wheeler, Crystal G; Ma, Tianlin; Harper, J Wade; Broker, Thomas R; Chow, Louise T

    2004-12-01

    Cyclin-dependent kinases (CDKs) play key roles in eukaryotic DNA replication and cell cycle progression. Phosphorylation of components of the preinitiation complex activates replication and prevents reinitiation. One mechanism is mediated by nuclear export of critical proteins. Human papillomavirus (HPV) DNA replication requires cellular machinery in addition to the viral replicative DNA helicase E1 and origin recognition protein E2. E1 phosphorylation by cyclin/CDK is critical for efficient viral DNA replication. We now show that E1 is phosphorylated by CDKs in vivo and that phosphorylation regulates its nucleocytoplasmic localization. We identified a conserved regulatory region for localization which contains a dominant leucine-rich nuclear export sequence (NES), the previously defined cyclin binding motif, three serine residues that are CDK substrates, and a putative bipartite nuclear localization sequence. We show that E1 is exported from the nucleus by a CRM1-dependent mechanism unless the NES is inactivated by CDK phosphorylation. Replication activities of E1 phosphorylation site mutations are reduced and correlate inversely with their increased cytoplasmic localization. Nuclear localization and replication activities of most of these mutations are enhanced or restored by mutations in the NES. Collectively, our data demonstrate that CDK phosphorylation controls E1 nuclear localization to support viral DNA amplification. Thus, HPV adopts and adapts the cellular regulatory mechanism to complete its reproductive program. PMID:15564503

  17. Role of the Zn2+ Motif of E1 in SUMO Adenylation*

    PubMed Central

    Wang, Jianghai; Chen, Yuan

    2010-01-01

    Post-translational modifications by ubiquitin-like proteins are among the most important mechanisms for regulating a wide variety of cellular functions. In these modifications an E1 enzyme activates each ubiquitin-like protein (Ubl) by adenylation of the Ubl C-terminal COOH group and then forms a thioester bond with the adenylated C-terminal COOH group of the Ubl. Previous x-ray crystallography studies revealed a conserved zinc motif in the SUMO and NEDD8 E1; however, the function of this Zn2+ motif is unclear. In this study, using quantitative ATP:PPi isotope exchange assays in combination with site-directed mutagenesis, we show that the conserved Zn2+ motif in the SUMO E1 is important for SUMO adenylation and is critical for the E1 pseudo-ordered substrate binding mechanism. Furthermore, Zn2+ motif mutants showed significantly reduced kcat values for ATP:PPi isotope exchange assays, suggesting that the Zn2+ motif is important in binding and preventing SUMO adenylate from dissociating from E1 before formation of the thioester conjugate. Because the Zn2+ motif is located in a cross-over loop that is known to have conformational flexibility, the results described here suggest that this cross-over loop interacts with Ubl in the multistep, dynamic process of Ubl activation by E1s. PMID:20501649

  18. Distinguishable promoter elements are involved in transcriptional activation by E1a and cyclic AMP.

    PubMed Central

    Lee, K A; Fink, J S; Goodman, R H; Green, M R

    1989-01-01

    The sequence motif CGTCA is critical for binding of a group of cellular transcription factors (ATF, CREB, E4F, and EivF) and for activation of certain E1a-inducible and cyclic AMP (cAMP)-inducible promoters. We have tested different promoter elements containing the CGTCA motif (referred to here as ATF-binding sites) for the ability to function as E1a or cAMP response elements. The adenovirus E4 promoter and the cellular vasoactive intestinal peptide (VIP) promoter responded differently to E1a and cAMP, demonstrating that the activating potential of ATF-binding sites within these promoters is not equivalent. While particular ATF-binding sites were sufficient for the activity of both the E4 (E1a inducibility) and VIP (cAMP inducibility) enhancers, these two enhancers had contrasting effects on E1a- and cAMP-inducible transcription. Thus, the relationship between E1a- and cAMP-inducible transcription is not simply explained by the action of these two inducers through the same promoter elements. Images PMID:2555692

  19. Strength of Chemical Bonds

    NASA Technical Reports Server (NTRS)

    Christian, Jerry D.

    1973-01-01

    Students are not generally made aware of the extraordinary magnitude of the strengths of chemical bonds in terms of the forces required to pull them apart. Molecular bonds are usually considered in terms of the energies required to break them, and we are not astonished at the values encountered. For example, the Cl2 bond energy, 57.00 kcal/mole, amounts to only 9.46 x 10(sup -20) cal/molecule, a very small amount of energy, indeed, and impossible to measure directly. However, the forces involved in realizing the energy when breaking the bond operate over a very small distance, only 2.94 A, and, thus, f(sub ave) approx. equals De/(r - r(sub e)) must be very large. The forces involved in dissociating the molecule are discussed in the following. In consideration of average forces, the molecule shall be assumed arbitrarily to be dissociated when the atoms are far enough separated so that the potential, relative to that of the infinitely separated atoms, is reduced by 99.5% from the potential of the molecule at the equilibrium bond length (r(sub e)) for Cl2 of 1.988 A this occurs at 4.928 A.

  20. Autophagy Protects against CYP2E1/Chronic Ethanol-Induced Hepatotoxicity

    PubMed Central

    Lu, Yongke; Cederbaum, Arthur I.

    2015-01-01

    Autophagy is an intracellular pathway by which lysosomes degrade and recycle long-lived proteins and cellular organelles. The effects of ethanol on autophagy are complex but recent studies have shown that autophagy serves a protective function against ethanol-induced liver injury. Autophagy was found to also be protective against CYP2E1-dependent toxicity in vitro in HepG2 cells which express CYP2E1 and in vivo in an acute alcohol/CYPE1-dependent liver injury model. The goal of the current report was to extend the previous in vitro and acute in vivo experiments to a chronic ethanol model to evaluate whether autophagy is also protective against CYP2E1-dependent liver injury in a chronic ethanol-fed mouse model. Wild type (WT), CYP2E1 knockout (KO) or CYP2E1 humanized transgenic knockin (KI), mice were fed an ethanol liquid diet or control dextrose diet for four weeks. In the last week, some mice received either saline or 3-methyladenine (3-MA), an inhibitor of autophagy, or rapamycin, which stimulates autophagy. Inhibition of autophagy by 3-MA potentiated the ethanol-induced increases in serum transaminase and triglyceride levels in the WT and KI mice but not KO mice, while rapamycin prevented the ethanol liver injury. Treatment with 3-MA enhanced the ethanol-induced fat accumulation in WT mice and caused necrosis in the KI mice; little or no effect was found in the ethanol-fed KO mice or any of the dextrose-fed mice. 3-MA treatment further lowered the ethanol-decrease in hepatic GSH levels and further increased formation of TBARS in WT and KI mice, whereas rapamycin blunted these effects of ethanol. Neither 3-MA nor rapamycin treatment affected CYP2E1 catalytic activity or content or the induction CYP2E1 by ethanol. The 3-MA treatment decreased levels of Beclin-1 and Atg 7 but increased levels of p62 in the ethanol-fed WT and KI mice whereas rapamycin had the opposite effects, validating inhibition and stimulation of autophagy, respectively. These results suggest that autophagy is protective against CYP2E1-dependent liver injury in a chronic ethanol-fed mouse model. We speculate that autophagy-dependent processes such as mitophagy and lipophagy help to minimize ethanol-induced CYP2E1-dependent oxidative stress and therefore the subsequent liver injury and steatosis. Attempts to stimulate autophagy may be helpful in lowering ethanol and CYP2E1-dependent liver toxicity. PMID:26501338

  1. Diabetes mellitus increases the in vivo activity of cytochrome P450 2E1 in humans

    PubMed Central

    Wang, Zaiqi; Hall, Stephen D; Maya, Juan F; Li, Lang; Asghar, Ali; Gorski, J C

    2003-01-01

    Aim Cytochrome P450 2E1 (CYP2E1) is thought to activate a number of protoxins, and has been implicated in the development of liver disease. Increased hepatic expression of CYP2E1 occurs in rat models of diabetes but it is unclear whether human diabetics display a similar up-regulation. This study was designed to test the hypothesis that human diabetics experience enhanced CYP2E1 expression. Methods The pharmacokinetics of a single dose of chlorzoxazone (500 mg), used as an index of hepatic CYP2E1 activity, was determined in healthy subjects (n = 10), volunteers with Type I (n = 13), and Type II (n = 8) diabetes mellitus. Chlorzoxazone and 6-hydroxychlorzoxazone in serum and urine were analysed by high-performance liquid chromatography. The expression of CYP2E1 mRNA in peripheral blood mononuclear cells was quantified by reverse transcriptase-polymerase chain reaction. Results The mean ± s.d. (90% confidence interval of the difference) chlorzoxazone area under the plasma concentration-time curve was significantly (P ≤ 0.05) reduced in obese Type II diabetics (15.7 ± 11.3 µg h ml−1; 9, 22) compared with healthy subjects (43.5 ± 16.9 µg h ml−1; 16, 40) and Type I diabetics (32.8 ± 9.2 µg h ml−1; 9, 25). There was a significant two-fold increase in the oral clearance of chlorzoxazone in obese Type II diabetics compared with healthy volunteers and Type I diabetics. The protein binding of chlorzoxazone was not significantly different between the three groups. In contrast, Type 1 diabetics and healthy volunteers demonstrated no difference in the oral clearance of chlorzoxazone. The urinary recovery of 6-hydroxychlorzoxazone as a percentage of the administered dose was not different between healthy, Type I and obese Type II diabetics. The elimination half-life of chlorzoxazone did not differ between the three groups. CYP2E1 mRNA was significantly elevated in Type I and obese Type II diabetics compared with healthy volunteers. The oral clearance of chlorzoxazone, elimination half-life, Tmax, and Cmax were not significantly influenced by weight, body mass index, serum glucose, serum cholesterol, or glycosylated haemoglobin. Conclusions There was a marked increase in hepatic CYP2E1 activity in obese Type II diabetics as assessed by chlorzoxazone disposition. Increased expression of CYP2E1 mRNA in peripheral blood mononuclear cells was found in both types of diabetes mellitus. Adverse hepatic events associated with Type II diabetes may be in part a result of enhanced CYP2E1 expression and activity. PMID:12534643

  2. CYP2E1 potentiates binge alcohol-induced gut leakiness, steatohepatitis, and apoptosis.

    PubMed

    Abdelmegeed, Mohamed A; Banerjee, Atrayee; Jang, Sehwan; Yoo, Seong-Ho; Yun, Jun-Won; Gonzalez, Frank J; Keshavarzian, Ali; Song, Byoung-Joon

    2013-12-01

    Ethanol-inducible cytochrome P450 2E1 (CYP2E1) contributes to increased oxidative stress and steatosis in chronic alcohol-exposure models. However, its role in binge ethanol-induced gut leakiness and hepatic injury is unclear. This study was aimed at investigating the role of CYP2E1 in binge alcohol-induced gut leakiness and the mechanisms of steatohepatitis. Female wild-type (WT) and Cyp2e1-null mice were treated with three doses of binge ethanol (WT-EtOH or Cyp2e1-null-EtOH) (6g/kg oral gavage at 12-h intervals) or dextrose (negative control). Intestinal histology of only WT-EtOH exhibited epithelial alteration and blebbing of lamina propria, and liver histology obtained at 6h after the last ethanol dose showed elevated steatosis with scattered inflammatory foci. These were accompanied by increased levels of serum endotoxin, hepatic enterobacteria, and triglycerides. All these changes, including the intestinal histology and hepatic apoptosis, determined by TUNEL assay, were significantly reversed when WT-EtOH mice were treated with the specific inhibitor of CYP2E1 chlormethiazole and the antioxidant N-acetylcysteine, both of which suppressed oxidative markers including intestinal CYP2E1. WT-EtOH also exhibited elevated amounts of serum TNF-?, hepatic cytokines, CYP2E1, and lipid peroxidation, with decreased levels of mitochondrial superoxide dismutase and suppressed aldehyde dehydrogenase 2 activity. Increased hepatocyte apoptosis with elevated levels of proapoptotic proteins and decreased levels of active (phosphorylated) p-AKT, p-AMPK, and peroxisome proliferator-activated receptor-?, all of which are involved in fat metabolism and inflammation, were observed in WT-EtOH. These changes were significantly attenuated in the corresponding Cyp2e1-null-EtOH mice. These data indicate that both intestinal and hepatic CYP2E1 induced by binge alcohol seems critical in binge alcohol-mediated increased nitroxidative stress, gut leakage, and endotoxemia; altered fat metabolism; and inflammation contributing to hepatic apoptosis and steatohepatitis. PMID:24064383

  3. Phenotyping of cytochrome P450 2E1 in vitro and in vivo.

    PubMed

    Ernstgård, Lena; Johanson, Gunnar; Karlsson, Anne-Sophie; Warholm, Margareta

    2007-06-01

    The aim of the present study was to develop and improve methods for phenotyping of CYP2E1, an important enzyme in the biotransformation of many industrial chemicals, therapeutic drugs and endogenous substances. The possibility to measure CYP2E1 activity in lymphocytes by using p-nitrophenol as a substrate and CYP2E1 protein levels by flow cytometry were studied in vitro. Further, the conventional chlorzoxazone method for in vivo phenotyping was studied by adjusting the dose to body weight in 10 healthy volunteers. Finally, the possibility to obtain the chlorzoxazone metabolic ratio in saliva samples was investigated. No CYP2E1 protein in lymphocytes was detected by using flow cytometry. Some enzyme activity was found in the experiments with p-nitrophenol, however, it could not be verified that it was catalyzed by CYP2E1. Chlorzoxazone and 6-hydroxychlorzoxazone were not detectable in saliva samples. The present in vivo experiments, combined with our previous data (in total 356 experiments in 50 subjects) show that the metabolic ratio increases with decreasing absorbed dose, expressed as the sum of chlorzoxazone and 6-hydroxychlorzoxazone in plasma at 2 h. The increase becomes pronounced at sum concentrations below 100 microM. In conclusion, chlorzoxazone metabolism in vivo remains the only available method for CYP2E1 phenotyping. The administered dose as well as the absorption of the probe influences the chlorzoxazone ratio. We suggest that a dose of 10 mg chlorzoxazone per kg body weight is used to estimate the CYP2E1 phenotype. Further, metabolic ratios should be disregarded if the sum of plasma chlorzoxazone and 6-hydroxychlorzoxazone is below 100 microM (blood sampled after 2 h). PMID:17584020

  4. Multifactorial Comparative Proteomic Study of Cytochrome P450 2E1 Function in Chronic Alcohol Administration

    PubMed Central

    Wang, Yuan; Kou, Yan; Wang, Xiaodong; Cederbaum, Arthur; Wang, Rong

    2014-01-01

    With the use of iTRAQ technique, a multifactorial comparative proteomic study can be performed. In this study, to obtain an overview of ethanol, CYP2E1 and gender effects on liver injury and gain more insight into the underlying molecular mechanism, mouse liver proteomes were quantitatively analyzed using iTRAQ under eight conditions including mice of different genders, wild type versus CYP2E1 knockout, and normal versus alcohol diet. A series of statistical and bioinformatic analyses were explored to simplify and clarify multifactorial comparative proteomic data. First, with the Principle Component analysis, six proteins, CYP2E1, FAM25, CA3, BHMT, HIBADH and ECHS1, involved in oxidation reduction, energy and lipid metabolism and amino acid metabolism, were identified as the most differentially expressed gene products across all of the experimental conditions of our chronic alcoholism model. Second, hierarchical clustering analysis showed CYP2E1 knockout played a primary role in the overall differential protein expression compared with ethanol and gender factors. Furthermore, pair-wise multiple comparisons have revealed that the only significant expression difference lied in wild-type and CYP2E1 knockout mice both treated with ethanol. Third, K-mean clustering analysis indicated that the CYP2E1 knockout had the reverse effect on ethanol induced oxidative stress and lipid oxidation. More importantly, IPA analysis of proteomic data inferred that the gene expressions of two upstream regulators, NRF2 and PPARα, regulated by chronic alcohol feeding and CYP2E1 knockout, are involved in ethanol induced oxidative stress and lipid oxidation. The present study provides an effectively comprehensive data analysis strategy to compare multiple biological factors, contributing to biochemical effects of alcohol on the liver. The mass spectrometry proteomics data have been deposited to the ProteomeXchange with data set identifier of PXD000635. PMID:24658151

  5. CYP2E1-dependent elevation of serum cholesterol, triglycerides, and hepatic bile acids by isoniazid.

    PubMed

    Cheng, Jie; Krausz, Kristopher W; Li, Feng; Ma, Xiaochao; Gonzalez, Frank J

    2013-01-15

    Isoniazid is the first-line medication in the prevention and treatment of tuberculosis. Isoniazid is known to have a biphasic effect on the inhibition-induction of CYP2E1 and is also considered to be involved in isoniazid-induced hepatotoxicity. However, the full extent and mechanism of involvement of CYP2E1 in isoniazid-induced hepatotoxicity remain to be thoroughly investigated. In the current study, isoniazid was administered to wild-type and Cyp2e1-null mice to investigate the potential toxicity of isoniazid in vivo. The results revealed that isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice, but produced elevated serum cholesterol and triglycerides, and hepatic bile acids in wild-type mice, as well as decreased abundance of free fatty acids in wild-type mice and not in Cyp2e1-null mice. Metabolomic analysis demonstrated that production of isoniazid metabolites was elevated in wild-type mice along with a higher abundance of bile acids, bile acid metabolites, carnitine and carnitine derivatives; these were not observed in Cyp2e1-null mice. In addition, the enzymes responsible for bile acid synthesis were decreased and proteins involved in bile acid transport were significantly increased in wild-type mice. Lastly, treatment of targeted isoniazid metabolites to wild-type mice led to similar changes in cholesterol, triglycerides and free fatty acids. These findings suggest that while CYP2E1 is not involved in isoniazid-induced hepatotoxicity, while an isoniazid metabolite might play a role in isoniazid-induced cholestasis through enhancement of bile acid accumulation and mitochondria ?-oxidation. PMID:23142471

  6. IL-4-mediated transcriptional regulation of human CYP2E1 by two independent signaling pathways.

    PubMed

    Wang, Jue; Hu, Yin; Nekvindova, Jana; Ingelman-Sundberg, Magnus; Neve, Etienne P A

    2010-11-15

    Cytochrome P450 2E1 (CYP2E1), the alcohol-inducible member of the cytochrome P450 super family, plays an important role in both physiological and pathophysiological processes. The present study focused on the induction of human CYP2E1 transcription by the anti-inflammatory cytokine interleukin-4 (IL-4) in human hepatoma B16A2 cells and revealed that this regulation is mediated by two independent pathways. RNA interference and overexpression of STAT6, indicated that the JAK-STAT signaling pathway is involved in IL-4-dependent induction and mutagenesis revealed the presence of a STAT6 binding site in CYP2E1 proximal promoter region (-583/-574-bp). However, inhibition of the JAK-STAT6 pathway using JAK1 siRNA constructs could only partially inhibit the induction of CYP2E1 promoter constructs indicating the presence of a second IL-4 responsive element. Indeed by using a series of truncated CYP2E1 promoter constructs a second more distal IL-4 responsive element (-1604/-1428-bp) was identified, which was further shown to involve the activation of IRS1/2. This induction was dependent on the transcription factor NFATc1 as IL-4-induced CYP2E1 expression was altered by silencing or overexpressing NFATc1. A NFATc1 binding site was identified in the second distal IL-4 responsive element (-1551/-1545-bp) by chromatin immunoprecipitation (ChIP) analysis. Finally simultaneous siRNA-mediated down-regulation of both STAT6 and NFATc1 or mutation of both STAT6 and NFATc1 binding sites abolished the IL-4-dependent transcriptional induction of CYP2E1, demonstrating that both pathways are required for maximal activation. In conclusion, the present study indicates that the induction of CYP2E1 transcription by IL-4 is mediated through two independent parallel pathways, involving JAK-STAT6 and IRS1/2 and NFATc1. PMID:20723539

  7. Epitope-mapped monoclonal antibodies against the HPV16E1--E4 protein.

    PubMed

    Doorbar, J; Ely, S; Coleman, N; Hibma, M; Davies, D H; Crawford, L

    1992-03-01

    The human papillomavirus (HPV) E1--E4 protein is the only nonstructural late protein encoded by the virus. We have isolated three hybridomas producing monoclonal antibodies to the E1--E4 protein of HPV16, which is the HPV type most frequently associated with cervical cancer. The three antibodies (TVG 401, 402, and 403) detect adjacent epitopes within the major seroreactive region of the molecule and show no reactivity against the E4 proteins of HPV1, HPV2, HPV4, or HPV6. The E1--E4 protein migrates as a 10K species on SDS-gel electrophoresis and forms cytoplasmic inclusion granules in infected cells in vitro similar in appearance to those produced by HPV1 in benign warts. In naturally occurring HPV16-induced tumors the E1--E4 protein was detected in the cytoplasm of cells in the upper layers of the lesion in areas in which HPV16 DNA replication was occurring, as determined by in situ hybridization. Although the epitopes recognized by these monoclonal antibodies survive brief fixation in 5% formaldehyde, reactivity was destroyed by prolonged fixation. These monoclonal antibodies represent the first against HPV16 E1--E4 and should complement those already available to E7 and L1 for the screening of frozen sections of clinical biopsies and will be of value in monitoring the progression of HPV infection from benign lesions to invasive cancer. PMID:1371027

  8. E1B and E4 oncoproteins of adenovirus antagonize the effect of apoptosis inducing factor

    SciTech Connect

    Turner, Roberta L.; Wilkinson, John C.; Ornelles, David A.

    2014-05-15

    Adenovirus inundates the productively infected cell with linear, double-stranded DNA and an abundance of single-stranded DNA. The cellular response to this stimulus is antagonized by the adenoviral E1B and E4 early genes. A mutant group C adenovirus that fails to express the E1B-55K and E4ORF3 genes is unable to suppress the DNA-damage response. Cells infected with this double-mutant virus display significant morphological heterogeneity at late times of infection and frequently contain fragmented nuclei. Nuclear fragmentation was due to the translocation of apoptosis inducing factor (AIF) from the mitochondria into the nucleus. The release of AIF was dependent on active poly(ADP-ribose) polymerase-1 (PARP-1), which appeared to be activated by viral DNA replication. Nuclear fragmentation did not occur in AIF-deficient cells or in cells treated with a PARP-1 inhibitor. The E1B-55K or E4ORF3 proteins independently prevented nuclear fragmentation subsequent to PARP-1 activation, possibly by altering the intracellular distribution of PAR-modified proteins. - Highlights: • E1B-55K or E4orf3 prevents nuclear fragmentation. • Nuclear fragmentation requires AIF and PARP-1 activity. • Adenovirus DNA replication activates PARP-1. • E1B-55K or E4orf3 proteins alter the distribution of PAR.

  9. Expression and Characterization of Acidothermus celluloyticus E1 Endoglucanase in Transgenic Duckweed Lemna minor 8627

    SciTech Connect

    Sun, Y.; Cheng, J. J.; Himmel, M. E.; Skory, C. D.; Adney, W. S.; Thomas, S. R.; Tisserat, B.; Nishimura, Y.; Yamamoto, Y. T.

    2007-01-01

    Endoglucanase E1 from Acidothermus cellulolyticus was expressed cytosolically under control of the cauliflower mosaic virus 35S promoter in transgenic duckweed, Lemna minor 8627 without any obvious observable phenotypic effects on morphology or rate of growth. The recombinant enzyme co-migrated with the purified catalytic domain fraction of the native E1 protein on western blot analysis, revealing that the cellulose-binding domain was cleaved near or in the linker region. The duckweed-expressed enzyme was biologically active and the expression level was up to 0.24% of total soluble protein. The endoglucanase activity with carboxymethylcellulose averaged 0.2 units mg protein{sup -1} extracted from fresh duckweed. The optimal temperature and pH for E1 enzyme activity were about 80 C and pH 5, respectively. While extraction with HEPES (N-[2-hydroxyethyl]piperazine-N{prime}-[2-ethanesulfonic acid]) buffer (pH 8) resulted in the highest recovery of total soluble proteins and E1 enzyme, extraction with citrate buffer (pH 4.8) at 65 C enriched relative amounts of E1 enzyme in the extract. This study demonstrates that duckweed may offer new options for the expression of cellulolytic enzymes in transgenic plants.

  10. Olive cultivar origin is a major cause of polymorphism for Ole e 1 pollen allergen

    PubMed Central

    Hamman-Khalifa, AbdelMounim; Castro, Antonio Jesús; Jiménez-López, José Carlos; Rodríguez-García, María Isabel; Alché, Juan de Dios

    2008-01-01

    Background Pollens from different olive (Olea europaea L.) cultivars have been shown to differ significantly in their content in Ole e 1 and in their overall allergenicity. This allergen is, in addition, characterized by a high degree of polymorphism in its sequence. The purpose of this study is to evaluate the putative presence of divergences in Ole e 1 sequences from different olive cultivars. Results RNA from pollen individually collected from 10 olive cultivars was used to amplify Ole e 1 sequences by RT-PCR, and the sequences were analyzed by using different bioinformatics tools. Numerous nucleotide substitutions were detected throughout the sequences, many of which resulted in amino acid substitutions in the deduced protein sequences. In most cases variability within a single variety was much lower than among varieties. Key amino acid changes in comparison with "canonical" sequences previously described in the literature included: a) the substitution of C19-relevant to the disulphide bond structure of the protein-, b) the presence of an additional N-glycosylation motif, and c) point substitutions affecting regions of Ole e 1 already described like relevant for the immunogenicity/allergenicity of the protein. Conclusion Varietal origin of olive pollen is a major factor determining the diversity of Ole e 1 variants. We consider this information of capital importance for the optimal design of efficient and safe allergen formulations, and useful for the genetic engineering of modified forms of the allergen among other applications. PMID:18218146

  11. DDX3 regulates cell growth through translational control of cyclin E1.

    PubMed

    Lai, Ming-Chih; Chang, Wen-Cheng; Shieh, Sheau-Yann; Tarn, Woan-Yuh

    2010-11-01

    DDX3 belongs to the DEAD box family of RNA helicases, but the details of its biological function remain largely unclear. Here we show that knockdown of DDX3 expression impedes G(1)/S-phase transition of the cell cycle. To know how DDX3 may act in cell cycle control, we screened for cellular mRNA targets of DDX3. Many of the identified DDX3 targets encoded cell cycle regulators, including G(1)/S-specific cyclin E1. DDX3 depletion specifically downregulates translation of cyclin E1 mRNA. Moreover, our data suggest that DDX3 participates in translation initiation of targeted mRNAs as well as in cell growth control via its RNA helicase activity. Consistent with these findings, we show that in the temperature-sensitive DDX3 mutant hamster cell line tsET24, cyclin E1 expression is downregulated at a nonpermissive temperature that inactivates mutant DDX3. Taken together, our results indicate that DDX3 is critical for translation of cyclin E1 mRNA, which provides an alternative mechanism for regulating cyclin E1 expression during the cell cycle. PMID:20837705

  12. Investigation of Indazole Unbinding Pathways in CYP2E1 by Molecular Dynamics Simulations

    PubMed Central

    Shen, Zhonghua; Cheng, Feixiong; Xu, You; Fu, Jing; Xiao, Wen; Shen, Jie; Liu, Guixia; Li, Weihua; Tang, Yun

    2012-01-01

    Human microsomal cytochrome P450 2E1 (CYP2E1) can oxidize not only low molecular weight xenobiotic compounds such as ethanol, but also many endogenous fatty acids. The crystal structure of CYP2E1 in complex with indazole reveals that the active site is deeply buried into the protein center. Thus, the unbinding pathways and associated unbinding mechanisms remain elusive. In this study, random acceleration molecular dynamics simulations combined with steered molecular dynamics and potential of mean force calculations were performed to identify the possible unbinding pathways in CYP2E1. The results show that channel 2c and 2a are most likely the unbinding channels of CYP2E1. The former channel is located between helices G and I and the B-C loop, and the latter resides between the region formed by the F-G loop, the B-C loop and the β1 sheet. Phe298 and Phe478 act as the gate keeper during indazole unbinding along channel 2c and 2a, respectively. Previous site-directed mutagenesis experiments also supported these findings. PMID:22442693

  13. CYP2E1 PstI polymorphism increases cervical neoplasia risk: a meta-analysis.

    PubMed

    Wang, X; He, Y

    2015-01-01

    Cytochrome P4502E1 (CYP2E1) is a key enzyme in the metabolic activation of many carcinogens, but the roles of CYP2E1 polymorphisms in cervical neoplasia (CN) are inconclusive. Published case-control cohort studies from the Pubmed, Embase, and China National Knowledge Infrastructure databases were retrieved. Data were extracted and pooled odds ratios with 95% confidence intervals were calculated. Seven studies examining 1097 cases and 1117 controls were included in this meta-analysis. The pooled effect size showed no association between CYP2E1 RsaI and DraI polymorphisms and CN risk in a codominant model. However, using a recessive model, an association between the PstI polymorphism and CN risk was observed (odds ratio: 2.10, 95% confidence interval: 0.96-4.62, P = 0.06), indicating that individuals with the homozygous rare genotype have a higher risk of developing CN compared to those with homozygous wild-type and heterozygous genotypes. When stratified by ethnicity, the PstI polymorphism was significantly correlated with CN susceptibility in non-Asians (odds ratio: 3.74, 95% confidence interval: 1.13-12.43, P = 0.03). This meta-analysis suggests that the CYP2E1 PstI polymorphism increases the risk of CN in non-Asians. PMID:26125714

  14. 2-Carboxyquinoxalines kill mycobacterium tuberculosis through noncovalent inhibition of DprE1.

    PubMed

    Neres, João; Hartkoorn, Ruben C; Chiarelli, Laurent R; Gadupudi, Ramakrishna; Pasca, Maria Rosalia; Mori, Giorgia; Venturelli, Alberto; Savina, Svetlana; Makarov, Vadim; Kolly, Gaelle S; Molteni, Elisabetta; Binda, Claudia; Dhar, Neeraj; Ferrari, Stefania; Brodin, Priscille; Delorme, Vincent; Landry, Valérie; de Jesus Lopes Ribeiro, Ana Luisa; Farina, Davide; Saxena, Puneet; Pojer, Florence; Carta, Antonio; Luciani, Rosaria; Porta, Alessio; Zanoni, Giuseppe; De Rossi, Edda; Costi, Maria Paola; Riccardi, Giovanna; Cole, Stewart T

    2015-03-20

    Phenotypic screening of a quinoxaline library against replicating Mycobacterium tuberculosis led to the identification of lead compound Ty38c (3-((4-methoxybenzyl)amino)-6-(trifluoromethyl)quinoxaline-2-carboxylic acid). With an MIC99 and MBC of 3.1 μM, Ty38c is bactericidal and active against intracellular bacteria. To investigate its mechanism of action, we isolated mutants resistant to Ty38c and sequenced their genomes. Mutations were found in rv3405c, coding for the transcriptional repressor of the divergently expressed rv3406 gene. Biochemical studies clearly showed that Rv3406 decarboxylates Ty38c into its inactive keto metabolite. The actual target was then identified by isolating Ty38c-resistant mutants of an M. tuberculosis strain lacking rv3406. Here, mutations were found in dprE1, encoding the decaprenylphosphoryl-d-ribose oxidase DprE1, essential for biogenesis of the mycobacterial cell wall. Genetics, biochemical validation, and X-ray crystallography revealed Ty38c to be a noncovalent, noncompetitive DprE1 inhibitor. Structure-activity relationship studies generated a family of DprE1 inhibitors with a range of IC50's and bactericidal activity. Co-crystal structures of DprE1 in complex with eight different quinoxaline analogs provided a high-resolution interaction map of the active site of this extremely vulnerable target in M. tuberculosis. PMID:25427196

  15. Strength Training for Young Athletes.

    ERIC Educational Resources Information Center

    Kraemer, William J.; Fleck, Steven J.

    This guide is designed to serve as a resource for developing strength training programs for children. Chapter 1 uses research findings to explain why strength training is appropriate for children. Chapter 2 explains some of the important physiological concepts involved in children's growth and development as they apply to developing strength…

  16. Strength Training and Your Child

    MedlinePlus

    ... All About Food Allergies Strength Training and Your Child KidsHealth > For Parents > Strength Training and Your Child Print A A A Text Size What's in ... muscles, and joints. Age Guidelines Generally, if your child is ready to participate in organized sports or ...

  17. CYP2E1 Potentiates Ethanol-induction of Hypoxia and HIF-1α in vivo

    PubMed Central

    Wang, Xiaodong; Wu, Defeng; Yang, Lili; Gan, Lixia; Cederbaum, Arthur I

    2013-01-01

    Ethanol induces hypoxia and elevates HIF-1α in the liver. CYP2E1 plays a role in the mechanisms by which ethanol generates oxidative stress, fatty liver and liver injury. The current study evaluated whether CYP2E1 contributes to ethanol-induced hypoxia and activation of HIF-1α in vivo and whether HIF-1α protects against or promotes CYP2E1-dependent toxicity in vitro. Wild type (WT), CYP2E1-knockin (KI) and CYP2E1 knockout (KO) mice were fed ethanol chronically; pair fed controls received isocaloric dextrose. Ethanol produced liver injury in the KI mice to a much greater extent than in the WT and KO mice. Protein levels of HIF-1α and downstream targets of HIF-1α activation were elevated in the ethanol-fed KI mice compared to the WT and KO mice. Levels of HIF prolylhydroxlase 2 which promotes HIF-1α degradation were decreased in the ethanol-fed KI mice in association with the increases in HIF-1α. Hypoxia occurred in the ethanol-fed CYP2E1 KI mice as shown by an increased area of staining using the hypoxia-specific marker pimonidazole. Hypoxia was lower in the ethanol-fed WT mice and lowest in the ethanol fed KO mice and all the dextrose-fed mice. In situ double staining showed that pimonidazole and CYP2E1 were co-localized to the same area of injury in the hepatic centrilobule. Increased protein levels of HIF-1α were also found after acute ethanol treatment of KI mice. Treatment of HepG2 E47 cells which express CYP2E1 with ethanol plus arachidonic (AA) acid or ethanol plus buthionine sulfoximine (BSO) which depletes GSH caused loss of cell viability to greater extent than in HepG2 C34 cells which do not express CYP2E1. These treatments elevated protein levels of HIF-1α to a greater extent in E47 cells than C34 cells. 2-Methoxyestradiol, an inhibitor of HIF-1α, blunted the toxic effects of ethanol plus AA and ethanol plus BSO in the E47 cells in association with inhibition of HIF-1α. The HIF-1α inhibitor also blocked the elevated oxidative stress produced by ethanol/AA or ethanol/BSO in the E47 cells. These results suggest that CYP2E1 plays a role in ethanol-induced hypoxia, oxidative stress and activation of HIF-1α and that HIF-1α contributes to CYP2E1-dependent ethanol-induced toxicity. Blocking HIF-1α activation and actions may have therapeutic implications for protection against ethanol/CYP2E1-induced oxidative stress, steatosis and liver injury. PMID:23669278

  18. A regulatory variant in CYP2E1 affects the risk of lung squamous cell carcinoma.

    PubMed

    Cao, Lei; Lin, Jia; He, Bing; Wang, Hongge; Rao, Juan; Liu, Yingwen; Zhang, Xuemei

    2014-01-01

    Cytochrome P450 2E1 (CYP2E1), an ethanol-inducible enzyme, has been shown to metabolically activate various carcinogens, which is critical for the development of cancers. It has been demonstrated that CYP2E1 polymorphisms alter the transcriptional activity. However, studies on the association between CYP2E1 -1239G>C polymorphism and non-small cell lung cancer have reported conflicting results. Thus, the gain of the present study was to investigate whether CYP2E1 -1239G>C polymorphism is associated with the development of non-small cell lung cancer in Chinese population. A case-control study was conducted in which CYP2E1 -1239G>C polymorphism was analyzed in 526 Chinese patients with non-small cell lung cancer and 526 age-matched healthy controls by polymerase chain reaction-restriction fragment length polymorphism. Odds ratios were estimated by multivariate logistic regression. A meta-analysis was conducted to evaluate the association of CYP2E1 -1239G>C polymorphism with the risk of lung cancer in Chinese population by calculating pooled odds ratio (OR). For CYP2E1 -1239G>C polymorphism, -1239C allele carriers (OR = 0.67; 95% confidence interval (CI) = 0.51-0.87; P = 0.002) were associated with a decreased risk of non-small cell lung cancer when compared with -1239GG homozygotes. In the group analyses by pathological types, for lung squamous cell carcinoma and other types, the ORs of the C allele carriers were 0.60 (95% CI = 0.41-0.88; P = 0.009) and 0.54 (95% CI = 0.30-0.99; P = 0.045). In the group analysis of smoking status, the OR for the -1239C allele-containing genotype was higher than that for -1239GG genotype (OR = 0.57; 95% CI = 0.40-0.81; P = 0.002) among smokers, but not among nonsmokers. Moreover, when the risk associated with CYP2E1 polymorphism was further evaluated within strata of <25 and ≥25 pack-years smoked, this effect between susceptible genotypes and smoking was mostly evident among light smokers (<25 pack-years) with OR of 0.42 (95% CI 0.23-0.79), but not among heavy smokers with OR of 0.87 (95% CI 0.53-1.43). In the group analyses by TNM stage, there was no significant difference between CYP2E1 -1239G>C polymorphism and the risk of non-small cell lung cancer. Meta-analysis data also showed that the carriers of CYP2E1 -1239C allele had a protect effect on the risk of lung cancer in Chinese with overall OR of 0.77 (95% CI 0.66-0.90). CYP2E1 -1239G>C polymorphism was associated with a decreased risk of development of non-small cell lung cancer in Chinese patients. The association displays a manner of gene-environment interaction between this polymorphism and smoking status. PMID:23934444

  19. Two distinct regions of the BPV1 E1 replication protein interact with the activation domain of E2.

    PubMed

    Moscufo, N; Sverdrup, F; Breiding, D E; Androphy, E J

    1999-12-15

    Papillomavirus E1 and E2 proteins co-operation in viral DNA replication is mediated by protein-protein interactions that lead to formation of an E1-E2 complex. To identify the domains involved, portions of the two proteins were expressed as fusions to the DNA-binding protein LexA or the transactivation domain of VP16 and analyzed by the yeast two-hybrid system. The C-terminal 266 amino acids of BPV1 E1 (E1C266) interacted strongly with E2 in the yeast system and in a mammalian two-hybrid assay. VP16-E1C266 interacted with a region encompassing amino acids 1-200 of the transactivation domain of E2 that was fused to LexA. The interaction between E1 full length and E2 was clearly observed only when E1 was expressed as LexA-E1 chimera. In addition, we found that in the LexA context also the N-terminal region encompassing the first 340 amino acids of E1 (E1N340) interacted with E2 full length. The interactions of E1N340 and E1C266 with E2 were confirmed also by in vitro binding studies. These observations demonstrate that two distinct regions of E1 mediate the interaction with E2 in vivo. PMID:10581387

  20. High-resolution study of extreme-ultraviolet emission from CO by electron impact: predissociation of the E ^1Π (v' = 0) state

    NASA Astrophysics Data System (ADS)

    Ciocca, M.; Kanik, I.; Ajello, J. M.

    1996-05-01

    We report a high-resolution study [0.0036 nm full width at half maximum (FWHM)] of electron-impact induced fluorescence spectra of CO for 30, 75 and 100 eV impact energies. At the specified resolution, now attainable with our newly constructed 3 m vacuum ultraviolet (vuv) spectrometer, we observe rotationally resolved emission, in the extreme-ultraviolet (euv), from the vibronic states B ^1Σ ^+(0), C ^1Σ ^+(0), and E ^1Π (0), to the ground state X ^1Σ ^+(0). A simple model of these bands, based on the Honl-London factors and unperturbed rotational constants, is shown to be in good agreement with the observed spectra. The predissociation yield of the the E ^1Π electronic state has been determined and accounted for in the model. In addition, the excitation function of the E ^1Π -> X ^1Σ ^+ transition, in the 0-800 eV impact energy range, has been measured for the first time, thus permitting determination of the oscillator strength by using a modified Born approximation analytical fit.

  1. Leinamycin E1 acting as an anticancer prodrug activated by reactive oxygen species.

    PubMed

    Huang, Sheng-Xiong; Yun, Bong-Sik; Ma, Ming; Basu, Hirak S; Church, Dawn R; Ingenhorst, Gudrun; Huang, Yong; Yang, Dong; Lohman, Jeremy R; Tang, Gong-Li; Ju, Jianhua; Liu, Tao; Wilding, George; Shen, Ben

    2015-07-01

    Leinamycin (LNM) is a potent antitumor antibiotic produced by Streptomyces atroolivaceus S-140, featuring an unusual 1,3-dioxo-1,2-dithiolane moiety that is spiro-fused to a thiazole-containing 18-membered lactam ring. Upon reductive activation in the presence of cellular thiols, LNM exerts its antitumor activity by an episulfonium ion-mediated DNA alkylation. Previously, we have cloned the lnm gene cluster from S. atroolivaceus S-140 and characterized the biosynthetic machinery responsible for the 18-membered lactam backbone and the alkyl branch at C3 of LNM. We now report the isolation and characterization of leinamycin E1 (LNM E1) from S. atroolivacues SB3033, a ΔlnmE mutant strain of S. atroolivaceus S-140. Complementary to the reductive activation of LNM by cellular thiols, LNM E1 can be oxidatively activated by cellular reactive oxygen species (ROS) to generate a similar episulfonium ion intermediate, thereby alkylating DNA and leading to eventual cell death. The feasibility of exploiting LNM E1 as an anticancer prodrug activated by ROS was demonstrated in two prostate cancer cell lines, LNCaP and DU-145. Because many cancer cells are under higher cellular oxidative stress with increased levels of ROS than normal cells, these findings support the idea of exploiting ROS as a means to target cancer cells and highlight LNM E1 as a novel lead for the development of anticancer prodrugs activated by ROS. The structure of LNM E1 also reveals critical new insights into LNM biosynthesis, setting the stage to investigate sulfur incorporation, as well as the tailoring steps that convert the nascent hybrid peptide-polyketide biosynthetic intermediate into LNM. PMID:26056295

  2. Inhibitory Potency of 4-Carbon Alkanes and Alkenes toward CYP2E1 Activity

    PubMed Central

    Hartman, Jessica H.; Miller, Grover P.; Boysen, Gunnar

    2016-01-01

    CYP2E1 has been implicated in the bioactivation of many small molecules into reactive metabolites which form adducts with proteins and DNA, and thus a better understanding of the molecular determinants of its selectivity are critical for accurate toxicological predictions. In this study, we determined the potency of inhibition of human CYP2E1 for various 4-carbon alkanes, alkenes and alcohols. In addition, known CYP2E1 substrates and inhibitors including 4-methylpyrazole, aniline, and dimethylnitrosamine were included to determine their relative potencies. Of the 1,3-butadiene-derived metabolites studied, 3,4-epoxy-1-butene was the strongest inhibitor with an IC50 of 110 μM compared to 1700 μM and 6600 μM for 1,2-butenediol and 1,2:3,4-diepoxybutane, respectively. Compared to known inhibitors, inhibitory potency of 3,4-epoxy-1-butene is between 4-methylpyrazole (IC50 = 1.8 μM) and dimethylnitrosamine (IC50 = 230 μM). All three butadiene metabolites inhibit CYP2E1 activity through a simple competitive mechanism. Among the 4-carbon compounds studied, the presence and location of polar groups seems to influence inhibitory potency. To further examine this notion, the investigation was extended to include structurally and chemically similar analogs, including propylene oxide and various butane alcohols. Those results demonstrated preferential recognition of CYP2E1 toward the type and location of polar and hydrophobic structural elements. Taken together, CYP2E1 metabolism may be modified in vivo by exposure to 4-carbon compounds, such as drugs, and nutritional constituents, a finding that highlights the complexity of exposure to mixtures. PMID:24561005

  3. Substitution of specific cysteine residues in E1 glycoprotein of classical swine fever virus strain Brescia affects formation of E1-E2 heterodimers and alters virulence in swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    E1, along with E^rns and E2, is one of the three envelope glycoproteins of Classical Swine Fever Virus (CSFV). E1 and E2 are anchored to the virus envelope at their carboxyl termini and E^rns loosely associates with the viral envelope. In infected cells, E2 forms homodimers and heterodimers with E1,...

  4. Influence of the adenovirus 5 E1A oncogene on chromatin remodelling.

    PubMed

    Mymryk, J S; Smith, M M

    1997-01-01

    In the eukaryotic nucleus, compaction of DNA into chromatin can limit the access of trans-acting factors, providing an additional level of regulation to processes such as transcription, replication, and repair. Recent studies have suggested that the protein products of the adenovirus 5 E1A oncogene can influence SWI-SNF and histone acetylase activities, two cellular processes that facilitate transcription in the context of chromatin. This review focuses on the unexpected effects of E1A on cellular processes that remodel chromatin in relation to its transcriptional and transforming activities. PMID:9250357

  5. CYP2E1-dependent elevation of serum cholesterol, triglycerides, and hepatic bile acids by isoniazid

    SciTech Connect

    Cheng, Jie; Krausz, Kristopher W.; Li, Feng; Ma, Xiaochao; Gonzalez, Frank J.

    2013-01-15

    Isoniazid is the first-line medication in the prevention and treatment of tuberculosis. Isoniazid is known to have a biphasic effect on the inhibition–induction of CYP2E1 and is also considered to be involved in isoniazid-induced hepatotoxicity. However, the full extent and mechanism of involvement of CYP2E1 in isoniazid-induced hepatotoxicity remain to be thoroughly investigated. In the current study, isoniazid was administered to wild-type and Cyp2e1-null mice to investigate the potential toxicity of isoniazid in vivo. The results revealed that isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice, but produced elevated serum cholesterol and triglycerides, and hepatic bile acids in wild-type mice, as well as decreased abundance of free fatty acids in wild-type mice and not in Cyp2e1-null mice. Metabolomic analysis demonstrated that production of isoniazid metabolites was elevated in wild-type mice along with a higher abundance of bile acids, bile acid metabolites, carnitine and carnitine derivatives; these were not observed in Cyp2e1-null mice. In addition, the enzymes responsible for bile acid synthesis were decreased and proteins involved in bile acid transport were significantly increased in wild-type mice. Lastly, treatment of targeted isoniazid metabolites to wild-type mice led to similar changes in cholesterol, triglycerides and free fatty acids. These findings suggest that while CYP2E1 is not involved in isoniazid-induced hepatotoxicity, while an isoniazid metabolite might play a role in isoniazid-induced cholestasis through enhancement of bile acid accumulation and mitochondria β-oxidation. -- Highlights: ► Isoniazid metabolites were elevated only in wild-type mice. ► Isoniazid caused no hepatotoxicity in wild-type and Cyp2e1-null mice. ► Isoniazid elevated serum cholesterol and triglycerides, and hepatic bile acids. ► Bile acid transporters were significantly decreased in isoniazid-treated mice.

  6. Melanoma cultures show different susceptibility towards E1A-, E1B-19 kDa- and fiber-modified replication-competent adenoviruses.

    PubMed

    Schmitz, M; Graf, C; Gut, T; Sirena, D; Peter, I; Dummer, R; Greber, U F; Hemmi, S

    2006-06-01

    Replicating adenovirus (Ad) vectors with tumour tissue specificity hold great promise for treatment of cancer. We have recently constructed a conditionally replicating Ad5 AdDeltaEP-TETP inducing tumour regression in a xenograft mouse model. For further improvement of this vector, we introduced four genetic modifications and analysed the viral cytotoxicity in a large panel of melanoma cell lines and patient-derived melanoma cells. (1) The antiapoptotic gene E1B-19 kDa (Delta19 mutant) was deleted increasing the cytolytic activity in 18 of 21 melanoma cells. (2) Introduction of the E1A 122-129 deletion (Delta24 mutant), suggested to attenuate viral replication in cell cycle-arrested cells, did not abrogate this activity and increased the cytolytic activity in two of 21 melanoma cells. (3) We inserted an RGD sequence into the fiber to extend viral tropism to alphav integrin-expressing cells, and (4) swapped the fiber with the Ad35 fiber (F35) enhancing the tropism to malignant melanoma cells expressing CD46. The RGD-fiber modification strongly increased cytolysis in all of the 11 CAR-low melanoma cells. The F35 fiber-chimeric vector boosted the cytotoxicity in nine of 11 cells. Our results show that rational engineering additively enhances the cytolytic potential of Ad vectors, a prerequisite for the development of patient-customized viral therapies. PMID:16482201

  7. Oscillator strengths and collision strengths for S v

    NASA Technical Reports Server (NTRS)

    Van Wyngaarden, W. L.; Henry, R. J. W.

    1981-01-01

    Observations of the optical extreme-ultraviolet spectrum of the Jupiter planetary system during the Voyager space mission revealed bright emission lines of some sulfur ions. The spectra of the torus at the orbit of Io are likely to contain S V lines. The described investigation provides oscillator strengths and collision strengths for the first four UV lines. The collision strengths from the ground state to four other excited states are also obtained. Use is made of a two-state calculation which is checked for convergence for some transitions by employing a three-state or a four-state approximation. Target wave functions for S V are calculated so that the oscillator strengths calculated in dipole length and dipole velocity approximations agree within 5%.

  8. Pelvic Muscle Strength After Childbirth

    PubMed Central

    Friedman, Sarah; Blomquist, Joan L.; Nugent, Joann M.; McDermott, Kelly C.; Muñoz, Alvaro; Handa, Victoria L.

    2013-01-01

    OBJECTIVE The objective was to estimate the effect of vaginal childbirth and other obstetric exposures on pelvic muscle strength 6–11 years after delivery and to investigate the relationship between pelvic muscle strength and pelvic floor disorders. METHODS Among 666 parous women, pelvic muscle strength was measured with a perineometer 6–11 years after delivery. Obstetric exposures were classified by review of hospital records. Pelvic floor outcomes, including stress incontinence, overactive bladder, anal incontinence, and prolapse symptoms, were assessed with a validated questionnaire. Pelvic organ support was assessed using the Pelvic Organ Prolapse Quantification system. Kruskal-Wallis tests were used to estimate the univariable associations of obstetric exposures and pelvic floor outcomes with peak muscle strength. Stepwise multivariable linear regression models were used to estimate the association between obstetric exposures and muscle strength. RESULTS In comparison with women who delivered all of their children by cesarean, peak muscle strength and duration of contraction were reduced among women with a history of vaginal delivery (39 compared with 29 cm H2O, P<.001). Pelvic muscle strength was further reduced after history of forceps delivery (17 cm H2O, P<.001). After vaginal delivery, reduced pelvic muscle strength was associated with symptoms of anal incontinence (P=.028) and pelvic organ prolapse on examination (P=.025); these associations were not observed among those who had delivered exclusively by cesarean. CONCLUSION Pelvic muscle strength almost a decade after childbirth is affected by vaginal delivery and by forceps delivery. Although statistically significant, some of the differences observed were small in magnitude. PMID:23090518

  9. 26 CFR 1.72(e)-1T - Treatment of distributions where substantially all contributions are employee contributions...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... all contributions are employee contributions (temporary). 1.72(e)-1T Section 1.72(e)-1T Internal... TAXES (CONTINUED) Items Specifically Included in Gross Income § 1.72(e)-1T Treatment of distributions... Reform Act (TRA) of 1984 change the law with regard to the treatment of non-annuity distributions...

  10. 26 CFR 1.72(e)-1T - Treatment of distributions where substantially all contributions are employee contributions...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... all contributions are employee contributions (temporary). 1.72(e)-1T Section 1.72(e)-1T Internal... TAXES (CONTINUED) Items Specifically Included in Gross Income § 1.72(e)-1T Treatment of distributions... Reform Act (TRA) of 1984 change the law with regard to the treatment of non-annuity distributions...

  11. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... § 1.1033(e)-1 Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of... applicable if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  12. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... § 1.1033(e)-1 Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of... applicable if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  13. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... § 1.1033(e)-1 Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of... applicable if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  14. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... § 1.1033(e)-1 Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of... applicable if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  15. Cytochrome P450 2E1 inhibition prevents hepatic carcinogenesis induced by diethylnitrosamine in alcohol-fed rats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chronic alcohol ingestion increases hepatic cytochrome P450 2E1 (CYP2E1), which is associated with hepatocarcinogenesis. We investigated whether treatment with chlormethiazole (CMZ), a CYP2E1 inhibitor, protects against alcohol-associated hepatic carcinogenesis in rats. Rats were fed either an ethan...

  16. 26 CFR 1.72(e)-1T - Treatment of distributions where substantially all contributions are employee contributions...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 72(o) are disregarded in their entirety (i.e., treated as neither employee contributions nor employer... all contributions are employee contributions (temporary). 1.72(e)-1T Section 1.72(e)-1T Internal... TAXES (CONTINUED) Items Specifically Included in Gross Income § 1.72(e)-1T Treatment of...

  17. 26 CFR 1.72(e)-1T - Treatment of distributions where substantially all contributions are employee contributions...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 72(o) are disregarded in their entirety (i.e., treated as neither employee contributions nor employer... all contributions are employee contributions (temporary). 1.72(e)-1T Section 1.72(e)-1T Internal... TAXES (CONTINUED) Items Specifically Included in Gross Income § 1.72(e)-1T Treatment of...

  18. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who... 40 Protection of Environment 5 2010-07-01 2010-07-01 false Designation Testing Checklist E Figure E-1 to Subpart E of Part 53 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

  19. Stability evaluation of a prostaglandin E1 saline solution packed in insulin syringes.

    PubMed

    Uebel, R A; Wium, C A; Schmidt, A C

    2001-02-01

    This article describes the preparation and stability of a sterile prostaglandin E1 solution. The solution is packed in insulin syringes as single dose injections for the treatment of erectile disfunction. The stability of this preparation is 24 weeks when stored at 5 degrees C. PMID:11313835

  20. Effects of Herbal Products and their Constituents on Human Cytochrome P4502E1 Activity

    PubMed Central

    Raner, Gregory M.; Cornelious, Sean; Moulick, Kamalika; Wang, Yingqing; Mortenson, Ashley; Cech, Nadja B.

    2010-01-01

    Ethanolic extracts from fresh Echinacea purpurea and Spilanthes acmella and dried Hydrastis canadensis were examined with regard to their ability to inhibit cytochrome P4502E1 mediated oxidation of p-nitrophenol in vitro. In addition, individual constituents of these extracts, including alkylamides from E. purpurea and S. acmella, caffeic acid derivatives from E. purpurea, and several of the major alkaloids from H. canadensis, were tested for inhibition using the same assay. H. canadensis (goldenseal) was a strong inhibitor of the P4502E1, and the inhibition appeared to be related to the presence of the alkaloids berberine, hydrastine and canadine in the extract. These compounds inhibited 2E1 with KI values ranging from 2.8 µM for hydrastine to 18 µM for berberine. The alkylamides present in E. purpurea and S. acmella also showed significant inhibition at concentrations as low as 25 µM, whereas the caffeic acid derivatives had no effect. Commercial green tea preparations, along with four of the individual tea catechins, were also examined and were found to have no effect on the activity of P4502E1. PMID:17658211

  1. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... HEART, LUNG, AND BLOOD INSTITUTE GRANTS FOR PREVENTION AND CONTROL PROJECTS § 52e.1 To what programs do... the prevention and control of heart, blood vessel, lung, and blood diseases, with special consideration given to the prevention and control of these diseases in children, and in populations that are...

  2. 26 CFR 1.1397E-1 - Qualified zone academy bonds.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... TAX (CONTINUED) INCOME TAXES (CONTINUED) Empowerment Zone Employment Credit § 1.1397E-1 Qualified zone... empowerment zone or enterprise community (as defined in section 1393), or there is a reasonable expectation... located in an empowerment zone or enterprise community for the entire term of the issue if the...

  3. 26 CFR 1.1397E-1T - Qualified zone academy bonds (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Empowerment Zone Employment Credit § 1.1397E-1T Qualified... empowerment zone or enterprise community (as defined in section 1393), or there is a reasonable expectation... located in an empowerment zone or enterprise community for the entire term of the issue if the...

  4. Defective human retinoblastoma protein identified by lack of interaction with the E1A oncoprotein.

    PubMed

    Paggi, M G; Martelli, F; Fanciulli, M; Felsani, A; Sciacchitano, S; Varmi, M; Bruno, T; Carapella, C M; Floridi, A

    1994-02-15

    Inactivating mutations of the retinoblastoma susceptibility gene (Rb) are involved in the pathogenesis of hereditary and sporadic retinoblastoma. Alterations in the Rb gene have also been found in several other human tumors occurring with epidemiological incidence higher than that of retinoblastoma. Four human malignant glioma cell lines were examined for abnormalities in the retinoblastoma gene product (pRb), using a procedure based on the interaction of pRb with an in vitro-translated adenovirus E1A oncoprotein. In the CRS-A2 cell line, derived from a glioblastoma multiforme, pRb did not bind with the in vitro-translated E1A protein. Restriction analysis of the CRS-A2 Rb gene and Rb mRNA expression provided patterns that could not be distinguished from the other glioma cell lines. Further investigation revealed the presence of a truncated pRb in the CRS-A2 cell line, due to a nucleotide insertion in the coding sequence at position 2550. In addition, this truncated Rb protein was undetectable in phosphorylated form. The binding assay with the in vitro-translated E1A was also used to study other cell lines with known mutations in the Rb gene. This method, which evaluates the interaction between in vitro-translated E1A and the pRb, is proposed as a rapid screening for detecting functional alterations in the retinoblastoma protein. PMID:8313367

  5. Induction of CYP2E1 in non-alcoholic fatty liver diseases.

    PubMed

    Aljomah, Ghanim; Baker, Susan S; Liu, Wensheng; Kozielski, Rafal; Oluwole, Janet; Lupu, Benita; Baker, Robert D; Zhu, Lixin

    2015-12-01

    Mounting evidence supports a contribution of endogenous alcohol metabolism in the pathogenesis of non-alcoholic steatohepatitis (NASH). However, it is not known whether the expression of alcohol metabolism genes is altered in the livers of simple steatosis. There is also a current debate on whether fatty acids induce CYP2E1 in fatty livers. In this study, expression of alcohol metabolizing genes in the liver biopsies of simple steatosis patients was examined by quantitative real-time PCR (qRT-PCR), in comparison to biopsies of NASH livers and normal controls. Induction of alcohol metabolizing genes was also examined in cultured HepG2 cells treated with ethanol or oleic acid, by qRT-PCR and Western blots. We found that the mRNA expression of alcohol metabolizing genes including ADH1C, ADH4, ADH6, catalase and CYP2E1 was elevated in the livers of simple steatosis, to similar levels found in NASH livers. In cultured HepG2 cells, ethanol induced the expression of CYP2E1 mRNA and protein, but not ADH4 or ADH6; oleic acid did not induce any of these genes. These results suggest that elevated alcohol metabolism may contribute to the pathogenesis of NAFLD at the stage of simple steatosis as well as more severe stages. Our in vitro data support that CYP2E1 is induced by endogenous alcohol but not by fatty acids. PMID:26551085

  6. Enhanced E1 transitions and {alpha}-clustering in {sup 212}Po

    SciTech Connect

    Suzuki, Y.; Ohkubo, S.

    2011-05-06

    An {alpha}+{sup 208}Pb(0{sup +},3{sup -}) cluster model explains the recently observed enhanced E1 transitions from the new negative-parity levels to the yrast states in {sup 212}Po. Heavy and light nuclei present good examples of surface clustering and well-localized clustering.

  7. E-1 Dynamic Fluid-Flow Model Update: EASY/ROCETS Enhancement and Model Development Support

    NASA Technical Reports Server (NTRS)

    Follett, Randolph F.; Taylor, Robert P.

    1998-01-01

    This report documents the research conducted to update computer models for dynamic fluid flow simulation of the E-1 test stand subsystems at te NASA John C. Stennis Space Center.Work also involved significant upgrades to the capabilities of EASY/ROCKETS library through the inclusion of the NIST-12 thermodynamic property database and development of new control system modules.

  8. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 18 2012-04-01 2012-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or...

  9. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 18 2014-04-01 2014-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or...

  10. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 18 2013-04-01 2013-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or...

  11. The 8-Pyrrole-Benzothiazinones Are Noncovalent Inhibitors of DprE1 from Mycobacterium tuberculosis

    PubMed Central

    Makarov, Vadim; Neres, João; Hartkoorn, Ruben C.; Ryabova, Olga B.; Kazakova, Elena; Šarkan, Michal; Huszár, Stanislav; Piton, Jérémie; Kolly, Gaëlle S.; Vocat, Anthony; Conroy, Trent M.; Mikušová, Katarína

    2015-01-01

    8-Nitro-benzothiazinones (BTZs), such as BTZ043 and PBTZ169, inhibit decaprenylphosphoryl-β-d-ribose 2′-oxidase (DprE1) and display nanomolar bactericidal activity against Mycobacterium tuberculosis in vitro. Structure-activity relationship (SAR) studies revealed the 8-nitro group of the BTZ scaffold to be crucial for the mechanism of action, which involves formation of a semimercaptal bond with Cys387 in the active site of DprE1. To date, substitution of the 8-nitro group has led to extensive loss of antimycobacterial activity. Here, we report the synthesis and characterization of the pyrrole-benzothiazinones PyrBTZ01 and PyrBTZ02, non-nitro-benzothiazinones that retain significant antimycobacterial activity, with MICs of 0.16 μg/ml against M. tuberculosis. These compounds inhibit DprE1 with 50% inhibitory concentration (IC50) values of <8 μM and present favorable in vitro absorption-distribution-metabolism-excretion/toxicity (ADME/T) and in vivo pharmacokinetic profiles. The most promising compound, PyrBTZ01, did not show efficacy in a mouse model of acute tuberculosis, suggesting that BTZ-mediated killing through DprE1 inhibition requires a combination of both covalent bond formation and compound potency. PMID:25987616

  12. NOVEL ASSAY TO ASSESS CYP-2E1-LIKE ACTIVITY IN THE JAPANESE MEDAKA (ORYZIAS LATIPES).

    EPA Science Inventory

    Liver microsomes and S-9 fraction of Japanese medaka (Oryzias latipes) metabolized the CYP2E1 specific substrate, p-nitrophenol (PNP), to a single hydroxylated product, 4-nitrocatechol. The use of liver S-9 fraction proved to be a viable alternative to liver microsomes and allowe...

  13. Coffee reduces SULT1E1 expression in human colon carcinoma Caco-2 cells.

    PubMed

    Isshiki, Marina; Ohta, Haruka; Tamura, Hiroomi

    2013-01-01

    Recent epidemiological studies have shown that moderate coffee consumption is associated with a lower risk of certain types of cancers, particularly colon cancer in postmenopausal women. To elucidate the molecular basis for the preventive action of coffee, we investigated the effect of coffee on estrogen sulfotransferase (SULT) because sulfation is the major pathway involved in the inactivation of estrogens. We found that coffee reduced SULT1E1 gene expression in human colon carcinoma Caco-2 cells. Treatment with 2.5% (v/v) coffee for 24 h resulted in a 60% reduction of the expression of the SULT1E1 gene in Caco-2 cells. Corresponding to reduced SULT1E1 gene expression, cytosolic estrogen SULT activity toward E(2) (20 nM) decreased by 25%. In addition, an accumulation of E(2) sulfates in the medium, which reflects cellular activity of estrogen SULT, decreased after the cells were treated with coffee. Major bioactive constituents in coffee such as caffeine, chlorogenic acid and caffeic acid did not show any effect. The inhibitory activity was extractable by using ethyl acetate. We also found that the inhibitory activity was produced by roasting the coffee beans. Mithramycin, an inhibitor of the transcription factor stimulating protein 1 (Sp1), was able to restore coffee-reduced SULT1E1 gene expression. Our data suggest that daily coffee consumption may reduce estrogen SULT activity, thereby enhancing estrogenic activity in the colon. PMID:23370358

  14. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Regulations 118 (26 CFR part 39, 1939 ed.) for the necessary amount. On January 1, 1965, B, a calendar year... 26 Internal Revenue 8 2012-04-01 2012-04-01 false Installment obligations transmitted at death... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In...

  15. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Regulations 118 (26 CFR part 39, 1939 ed.) for the necessary amount. On January 1, 1965, B, a calendar year... 26 Internal Revenue 8 2014-04-01 2014-04-01 false Installment obligations transmitted at death... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In...

  16. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Regulations 118 (26 CFR part 39, 1939 ed.) for the necessary amount. On January 1, 1965, B, a calendar year... 26 Internal Revenue 8 2011-04-01 2011-04-01 false Installment obligations transmitted at death... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In...

  17. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Regulations 118 (26 CFR part 39, 1939 ed.) for the necessary amount. On January 1, 1965, B, a calendar year... 26 Internal Revenue 8 2013-04-01 2013-04-01 false Installment obligations transmitted at death... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In...

  18. 26 CFR 301.6501(e)-1T - Omission from return (temporary).

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... and Collection § 301.6501(e)-1T Omission from return (temporary). (a) Income taxes—(1) General rule. (i) If the taxpayer omits from the gross income stated in the return of a tax imposed by subtitle A... the gross income so stated, the tax may be assessed, or a proceeding in court for the collection...

  19. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 18 2011-04-01 2011-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or...

  20. 26 CFR 301.6511(e)-1 - Special rules applicable to manufactured sugar.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Special rules applicable to manufactured sugar... Assessment and Collection § 301.6511(e)-1 Special rules applicable to manufactured sugar. (a) Use as... the person entitled thereto. Such right accrues as of the date the manufactured sugar, or...

  1. 26 CFR 301.6229(e)-1 - Information with respect to unidentified partner.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 CFR part 1, revised April 1, 2001. ... 26 Internal Revenue 18 2014-04-01 2014-04-01 false Information with respect to unidentified....6229(e)-1 Information with respect to unidentified partner. (a) In general. A partner who is...

  2. 26 CFR 301.6229(e)-1 - Information with respect to unidentified partner.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 CFR part 1, revised April 1, 2001. ... 26 Internal Revenue 18 2012-04-01 2012-04-01 false Information with respect to unidentified....6229(e)-1 Information with respect to unidentified partner. (a) In general. A partner who is...

  3. 26 CFR 301.6229(e)-1 - Information with respect to unidentified partner.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 CFR part 1, revised April 1, 2001. ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Information with respect to unidentified....6229(e)-1 Information with respect to unidentified partner. (a) In general. A partner who is...

  4. 26 CFR 301.6229(e)-1 - Information with respect to unidentified partner.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 CFR part 1, revised April 1, 2001. ... 26 Internal Revenue 18 2013-04-01 2013-04-01 false Information with respect to unidentified....6229(e)-1 Information with respect to unidentified partner. (a) In general. A partner who is...

  5. 26 CFR 301.6229(e)-1 - Information with respect to unidentified partner.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 CFR part 1, revised April 1, 2001. ... 26 Internal Revenue 18 2011-04-01 2011-04-01 false Information with respect to unidentified....6229(e)-1 Information with respect to unidentified partner. (a) In general. A partner who is...

  6. Benzene metabolism by human liver microsomes in relation to cytochrome P450 2E1 activity.

    PubMed

    Seaton, M J; Schlosser, P M; Bond, J A; Medinsky, M A

    1994-09-01

    Low levels of benzene from sources including cigarette smoke and automobile emissions are ubiquitous in the environment. Since the toxicity of benzene probably results from oxidative metabolites, an understanding of the profile of biotransformation of low levels of benzene is critical in making a valid risk assessment. To that end, we have investigated metabolism of a low concentration of [14C]benzene (3.4 microM) by microsomes from human, mouse and rat liver. The extent of phase I benzene metabolism by microsomal preparations from 10 human liver samples and single microsomal preparations from both mice and rats was then related to measured activities of cytochrome P450 (CYP) 2E1. Measured CYP 2E1 activities, as determined by hydroxylation of p-nitrophenol, varied 13-fold (0.253-3.266 nmol/min/mg) for human samples. The fraction of benzene metabolized in 16 min ranged from 10% to 59%. Also at 16 min, significant amounts of oxidative metabolites were formed. Phenol was the main metabolite formed by all but two human microsomal preparations. In those samples, both of which had high CYP 2E1 activity, hydroquinone was the major metabolite formed. Both hydroquinone and catechol formation showed a direct correlation with CYP 2E1 activity over the range of activities present. A simulation model was developed based on a mechanism of competitive inhibition between benzene and its oxidized metabolites, and was fit to time-course data for three human liver preparations. Model calculations for initial rates of benzene metabolism ranging from 0.344 to 4.442 nmol/mg/min are directly proportional to measured CYP 2E1 activities. The model predicted the dependence of benzene metabolism on the measured CYP 2E1 activity in human liver samples, as well as in mouse and rat liver samples. These results suggest that differences in measured hepatic CYP 2E1 activity may be a major factor contributing to both interindividual and interspecies variations in hepatic metabolism of benzene. Validation of this system in vivo should lead to more accurate assessment of the risk of benzene's toxicity following low-level exposure. PMID:7923572

  7. Specific estrogen sulfotransferase (SULT1E1) substrates and molecular imaging probe candidates

    PubMed Central

    Cole, Graham B.; Keum, Gyochang; Liu, Jie; Small, Gary W.; Satyamurthy, Nagichettiar; Kepe, Vladimir; Barrio, Jorge R.

    2010-01-01

    This work focuses on the development of specific substrates for estrogen sulfotransferase (SULT1E1) to produce molecular imaging probes for this enzyme. SULT1E1 is a key enzyme in estrogen homeostasis, playing a central role in the prevention and development of human disease. In vitro sulfation assays showed alkyl and aryl substitutions to a fused heterocyclic system modeled after β-naphthol (βN), based on compounds that interact with the estrogen receptor, rendered several molecules with enhanced specificity for SULT1E1 over SULT1A1*1, SULT1A1*2, SULT1A3, and SULT2A1. Several 6-hydroxy-2-arylbenzothiazoles tested demonstrated excellent affinity—Vmax/Km ratios—and specificity for SULT1E1. Km values ranged from 0.12–2.36 μM. A strong correlation was observed between polarity of the 4′-sustituent on the 2-aryl moiety (Hammett σp) and the log(Vmax/Km) (r = 0.964). Substrate sensitivity is influenced by the acidity of the 6-phenolic group demonstrated by correlating its 1H NMR chemical shift (δOH) with the log(Vmax/Km) (r = 0.963). Acidity is mediated by the electron withdrawing capacity of the 4′-substituent outlined by the correlation of the C-2 13C NMR chemical shift (δC2) with the log(Vmax/Km) (r = 0.987). 2-[4-(Methylamino)phenyl]-6-hydroxybenzothiazole (2b) was radiolabeled with carbon-11 (11C-(2b)) and used in vivo for microPET scanning and tissue metabolite identification. High PET signal was paralleled with the presence of radiolabeled 11C-(2b)-6-O-sulfate and the SULT1E1 protein detected by western blot. Because this and other members of this family presenting specificity for SULT1E1 can be labeled with carbon-11 or fluorine-18, in vivo assays of SULT1E1 functional activity are now feasible in humans. PMID:20304798

  8. Osteogenic gene expression of murine osteoblastic (MC3T3-E1) cells under cyclic tension

    NASA Astrophysics Data System (ADS)

    Kao, C. T.; Chen, C. C.; Cheong, U.-I.; Liu, S. L.; Huang, T. H.

    2014-08-01

    Low-level laser therapy (LLLT) can promote cell proliferation. The remodeling ability of the tension side of orthodontic teeth affects post-orthodontic stability. The purpose of the present study was to investigate the osteogenic effects of LLLT on osteoblast-like cells treated with a simulated tension system that provides a mechanical tension regimen. Murine osteoblastic (MC3T3-E1) cells were cultured in a Flexcell strain unit with programmed loads of 12% elongation at a frequency of 0.5 Hz for 24 and 48 h. The cultured cells were treated with a low-level diode laser using powers of 5 J and 10 J. The proliferation of MC3T3-E1 cells was determined using the Alamar Blue assay. The expression of osteogenic genes (type I collagen (Col-1), osteopontin (OPN), osteocalcin (OC), osteoprotegerin (OPG), receptor activator of nuclear factor kappa B ligand (RANKL), bone morphologic protein (BMP-2), and bone morphologic protein (BMP-4)) in MC3T3-E1 cells was analyzed using reverse transcription polymerase chain reaction (RT-PCR). The data were analyzed using one-way analysis of variance. The proliferation rate of tension-cultured MC3T3-E1 cells under 5 J and 10 J LLLT increased compared with that of the control group (p < 0.05). Prominent mineralization of the MC3T3-E1 cells was visible using a von Kossa stain in the 5 J LLLT group. Osteogenic genes (Col-1, OC, OPG and BMP-2) were significantly expressed in the MC3T3-E1 cells treated with 5 J and 10 J LLLT (p < 0.05). LLLT in tension-cultured MC3T3-E1 cells showed synergistic osteogenic effects, including increases in cell proliferation and Col-1, OPN, OC, OPG and BMP-2 gene expression. LLLT might be beneficial for bone remodeling on the tension side of orthodontics.

  9. Information Content of the Low-Energy Electric Dipole Strength: Correlation Analysis

    SciTech Connect

    Reinhard, P.-G.; Nazarewicz, Witold

    2013-01-01

    Background: Recent experiments on the electric dipole (E1) polarizability in heavy nuclei have stimulated theoretical interest in the low-energy electric dipole strength, both isovector and isoscalar. Purpose: We study the information content carried by the electric dipole strength with respect to isovector and isoscalar indicators characterizing bulk nuclear matter and finite nuclei. To separate isoscalar and isovector modes, and low-energy strength and giant resonances, we analyze the E1 strength as a function of the excitation energy E and momentum transfer q. Methods: We use the self-consistent nuclear density functional theory with Skyrme energy density functionals, augmented by the random phase approximation, to compute the E1 strength and covariance analysis to assess correlations between observables. Calculations are performed for the spherical, doubly magic nuclei 208Pb and 132Sn. Results: We demonstrate that E1 transition densities in the low-energy region below the giant dipole resonance exhibit appreciable state dependence and multinodal structures, which are fingerprints of weak collectivity. The correlation between the accumulated low-energy strength and the symmetry energy is weak, and dramatically depends on the energy cutoff assumed. On the other hand, a strong correlation is predicted between isovector indicators and the accumulated isovector strength at E around 20 MeV and momentum transfer q 0.65 fm 1. Conclusions: Momentum- and coordinate-space patterns of the low-energy dipole modes indicate a strong fragmentation into individual particle-hole excitations. The global measure of low-energy dipole strength correlates poorly with the nuclear symmetry energy and other isovector characteristics. Consequently, our results do not support the suggestion that there exists a collective pygmy dipole resonance, which is a strong indicator of nuclear isovector properties. By considering nonzero values of momentum transfer, one can isolate individual excitations and nicely separate low-energy excitations from the T=1 and T=0 giant collective modes. That is, measurements at q>0 may serve as a tool to correlate the E1 strength with certain bulk observables, such as incompressibility and symmetry energy.

  10. Structural basis of inhibition of Mycobacterium tuberculosis DprE1 by benzothiazinone inhibitors

    PubMed Central

    Batt, Sarah M.; Jabeen, Talat; Bhowruth, Veemal; Quill, Lee; Lund, Peter A.; Eggeling, Lothar; Alderwick, Luke J.; Fütterer, Klaus; Besra, Gurdyal S.

    2012-01-01

    Resistance against currently used antitubercular therapeutics increasingly undermines efforts to contain the worldwide tuberculosis (TB) epidemic. Recently, benzothiazinone (BTZ) inhibitors have shown nanomolar potency against both drug-susceptible and multidrug-resistant strains of the tubercle bacillus. However, their proposed mode of action is lacking structural evidence. We report here the crystal structure of the BTZ target, FAD-containing oxidoreductase Mycobacterium tuberculosis DprE1, which is essential for viability. Different crystal forms of ligand-free DprE1 reveal considerable levels of structural flexibility of two surface loops that seem to govern accessibility of the active site. Structures of complexes with the BTZ-derived nitroso derivative CT325 reveal the mode of inhibitor binding, which includes a covalent link to conserved Cys387, and reveal a trifluoromethyl group as a second key determinant of interaction with the enzyme. Surprisingly, we find that a noncovalent complex was formed between DprE1 and CT319, which is structurally identical to CT325 except for an inert nitro group replacing the reactive nitroso group. This demonstrates that binding of BTZ-class inhibitors to DprE1 is not strictly dependent on formation of the covalent link to Cys387. On the basis of the structural and activity data, we propose that the complex of DrpE1 bound to CT325 is a representative of the BTZ-target complex. These results mark a significant step forward in the characterization of a key TB drug target. PMID:22733761

  11. CYP2E1 and NQO1 genotypes and bladder cancer risk in a Lebanese population

    PubMed Central

    Basma, Hussein A; Kobeissi, Loulou H; Jabbour, Michel E; Moussa, Mohamad A; Dhaini, Hassan R

    2013-01-01

    Urinary bladder cancer incidence in Lebanon ranks among the highest in the world. Cytochrome P450 2E1 (CYP2E1), NAD(P)H quinone oxidoreductase1 (NQO1), and N-Acetyltransferase1 (NAT1), are drug-metabolizing enzymes (DMEs) involved in the metabolism of carcinogens, such as arylamines and heterocyclic amines, implicated in bladder cancer. The present study attempts to investigate the role of these DMEs genetic polymorphism in bladder cancer risk among Lebanese men. 54 cases and 106 controls were recruited from two hospitals in Beirut. An interview-based questionnaire was administered to assess suspected environmental and occupational risk factors. PCR-RFLP was performed on blood-based DNA samples to determine DMEs genotypes. Associations between bladder cancer and putative risk factors were measured using adjusted odds ratios (ORs) and their 95% confidence intervals (CIs). Results showed CYP2E1 c1/c1, NAT1*14A, and smoking, to be risk factors for bladder cancer. No significant differences in frequency distribution of the NQO1 genotypes were found in cases versus controls. The odds of carrying the CYP2E1 c1/c1 genotype were 4 times higher in cases compared to controls (OR=3.97, 95% CI: 0.48-32.7). The odds of carrying at least one NAT1*14A allele were 14 times higher in cases versus controls (OR=14.4, 95% CI: 1.016-204.9). Our study suggests CYP2E1 c1/c1, NAT1*14A, and smoking, as potential risk factors for bladder cancer in Lebanese. Further studies with larger samples must be conducted to confirm these findings. PMID:24319536

  12. Earthquake stress drop and laboratory-inferred interseismic strength recovery

    USGS Publications Warehouse

    Beeler, N.M.; Hickman, S.H.; Wong, T.-F.

    2001-01-01

    We determine the scaling relationships between earthquake stress drop and recurrence interval tr that are implied by laboratory-measured fault strength. We assume that repeating earthquakes can be simulated by stick-slip sliding using a spring and slider block model. Simulations with static/kinetic strength, time-dependent strength, and rate- and state-variable-dependent strength indicate that the relationship between loading velocity and recurrence interval can be adequately described by the power law VL ??? trn, where n=-1. Deviations from n=-1 arise from second order effects on strength, with n>-1 corresponding to apparent time-dependent strengthening and n<-1 corresponding to weakening. Simulations with rate and state-variable equations show that dynamic shear stress drop ????d scales with recurrence as d????d/dlntr ??? ??e(b-a), where ??e is the effective normal stress, ??=??/??e, and (a-b)=d??ss/dlnV is the steady-state slip rate dependence of strength. In addition, accounting for seismic energy radiation, we suggest that the static shear stress drop ????s scales as d????s/dlntr ??? ??e(1+??)(b-a), where ?? is the fractional overshoot. The variation of ????s with lntr for earthquake stress drops is somewhat larger than implied by room temperature laboratory values of ?? and b-a. However, the uncertainty associated with the seismic data is large and the discrepancy between the seismic observations and the rate of strengthening predicted by room temperature experiments is less than an order of magnitude. Copyright 2001 by the American Geophysical Union.

  13. Heterologous expression of human cytochrome P450 2E1 in HepG2 cell line

    PubMed Central

    Zhuge, Jian; Luo, Ye; Yu, Ying-Nian

    2003-01-01

    AIM: Human cytochrome P-450 2E1 (CYP2E1) takes part in the biotransformation of ethanol, acetone, many small-molecule substrates and volatile anesthetics. CYP2E1 is involved in chemical activation of many carcinogens, procarcinogens, and toxicants. To assess the metabolic and toxicological characteristics of CYP2E1, we cloned CYP2E1 cDNA and established a HepG2 cell line stably expressing recombinant CYP 2E1. METHODS: Human CYP2E1 cDNA was amplified with reverse transcription-polymerase chain reaction (RT-PCR) from total RNAs extracted from human liver and cloned into pGEM-T vector. The cDNA segment was identified by DNA sequencing and subcloned into a mammalian expression vector pREP9. A transgenic cell line was established by transfecting the recombinant plasmid of pREP9-CYP2E1 to HepG2 cells. The expression of CYP2E1 mRNA was validated by RT-PCR. The enzyme activity of CYP2E1 catalyzing oxidation of 4-nitrophenol in postmitochondrial supernate (S9) fraction of the cells was determined by spectrophotometry. The metabolic activation of HepG2-CYP2E1 cells was assayed by N-nitrosodiethylamine (NDEA) cytotoxicity and micronucleus test. RESULTS: The cloned CYP2E1 cDNA segment was identical to that reported by Umeno et al (GenBank access No. J02843). HepG2-CYP2E1 cells expressed CYP2E1 mRNA and had 4-nitrophenol hydroxylase activity (0.162 ± 0.025 nmol·min-1·mg-1 S9 protein), which were undetectable in parent HepG2 cells. HepG2-CYP2E1 cells increased the cytotoxicity and micronucleus rate of NDEA in comparison with those of HepG2 cells. CONCLUSION: The cDNA of human CYP2E1 can be successfully cloned, and a cell line, HepG2-CYP2E1, which can efficiently express mRNA and has CYP2E1 activity, is established. The cell line is useful for testing the cytotoxicity, mutagenicity and metabolism of xenobiotics, which may possibly be activated or metabolized by CYP2E1. PMID:14669323

  14. 2E1 Ar(17+) decay and conventional radioactive sources to determine efficiency of semiconductor detectors.

    PubMed

    Lamour, Emily; Prigent, Christophe; Eberhardt, Benjamin; Rozet, Jean Pierre; Vernhet, Dominique

    2009-02-01

    Although reliable models may predict the detection efficiency of semiconductor detectors, measurements are needed to check the parameters supplied by the manufacturers, namely, the thicknesses of dead layer, beryllium window, and crystal active area. The efficiency of three silicon detectors has been precisely investigated in their entire photon energy range of detection. In the zero to a few keV range, we developed a new method based on the detection of the 2E1 decay of the metastable Ar(17+) 2s-->1s transition. Very good theoretical knowledge of the energetic distribution of the 2E1 decay mode enables precise characterization of the absorbing layers in front of the detectors. In the high-energy range (>10 keV), the detector crystal thickness plays a major role in the detection efficiency and has been determined using a (241)Am source. PMID:19256636

  15. 2E1 Ar17+ decay and conventional radioactive sources to determine efficiency of semiconductor detectors

    NASA Astrophysics Data System (ADS)

    Lamour, Emily; Prigent, Christophe; Eberhardt, Benjamin; Rozet, Jean Pierre; Vernhet, Dominique

    2009-02-01

    Although reliable models may predict the detection efficiency of semiconductor detectors, measurements are needed to check the parameters supplied by the manufacturers, namely, the thicknesses of dead layer, beryllium window, and crystal active area. The efficiency of three silicon detectors has been precisely investigated in their entire photon energy range of detection. In the zero to a few keV range, we developed a new method based on the detection of the 2E1 decay of the metastable Ar17+ 2s→1s transition. Very good theoretical knowledge of the energetic distribution of the 2E1 decay mode enables precise characterization of the absorbing layers in front of the detectors. In the high-energy range (>10 keV), the detector crystal thickness plays a major role in the detection efficiency and has been determined using a A241m source.

  16. Association between the CYP2E1 polymorphisms and lung cancer risk: a meta-analysis.

    PubMed

    Ye, Xiang-Hua; Song, Liang; Peng, Ling; Bu, Zhibin; Yan, Sen-Xiang; Feng, Jie; Zhu, Xin-Li; Liao, Xin-Biao; Yu, Xue-Lin; Yan, Danfang

    2015-04-01

    The previous, published data on the association between CYP2E1 RsaI (rs2031920), DraI (rs6413432) polymorphisms and lung cancer risk remained controversial. Hence, we performed a meta-analysis to investigate the association between lung cancer and CYP2E1 RsaI (5,074 cases and 6,828 controls from 34 studies), and CYP2E1 DraI (2,093 cases and 2,508 controls from 16 studies) in different inheritance models. Overall, significantly decreased lung cancer risk was observed (dominant model: odds ratio (OR) 0.80, 95 % confidence interval (95 % CI) 0.71-0.90; heterozygote model: OR 0.80, 95 % CI 0.70-0.90; additive model: OR 0.82, 95 % CI 0.72-0.94) when all the eligible studies were pooled into the meta-analysis of CYP2E1 RsaI polymorphism. In further stratified and sensitivity analyses, significantly decreased lung cancer risk was found among Asians (dominant model: OR 0.81, 95 % CI 0.71-0.93; heterozygous model: OR 0.81, 95 % CI 0.69-0.95), population-based studies (dominant model: OR 0.69, 95 % CI 0.54-0.88; recessive model: OR 0.39, 95 % CI 0.16-0.91; additive model: OR 0.67, 95 % CI 0.53-0.84; homozygous model: OR 0.34, 95 % CI 0.14-0.80; heterozygous model: OR 0.70, 95 % CI 0.54-0.91), hospital-based studies (dominant model: OR 0.80, 95 % CI 0.69-0.93; additive model: OR 0.84, 95 % CI 0.70-1.00; heterozygous model: OR 0.80, 95 % CI 0.68-0.95), lung AC (heterozygous model: OR 0.84, 95 % CI 0.71-1.00), smokers (dominant model: OR 0.72, 95 % CI 0.55-0.94), and non-smokers (dominant model: OR 0.74, 95 % CI 0.61-0.91). There was no significant association between CYP2E1 DraI polymorphism and the risk of lung cancer when all the eligible studies were pooled into the meta-analysis. However, in further stratified and sensitivity analyses, significant association was observed among smokers (dominant model: OR 0.49, 95 % CI 0.35-0.69). In summary, this meta-analysis indicates that CYP2E1 RsaI polymorphism is associated with lung cancer risk among Asians, CYP2E1 RsaI polymorphism may be associated with lung adenocarcinoma risk, and CYP2E1 RsaI and DraI polymorphisms may be associated with decreased lung cancer risk in smokers. PMID:25336053

  17. Phylogenetic analysis of envelope glycoprotein (E1) gene of rubella viruses prevalent in Japan in 2004.

    PubMed

    Saitoh, Mika; Shinkawa, Naomi; Shimada, Shinichi; Segawa, Yukari; Sadamasu, Kenji; Hasegawa, Michiya; Kato, Masahiko; Kozawa, Kunihisa; Kuramoto, Tsuyoshi; Nishio, Osamu; Kimura, Hirokazu

    2006-01-01

    We performed a molecular epidemiological study on the envelope glycoprotein gene (E1 gene) obtained by PCR amplification from specimens of 17 rubella patients in certain areas (Gunma, Saitama, and Kagoshima prefectures, and Tokyo metropolitan area) in Japan in 2004. In these sequences of partially amplified DNAs (283 bases) within the E1 gene, no nucleotide substitution was observed. They were classified into genotype 1D of clade 1 in the constructed phylogenetic tree. One amino acid substitution was found between the amino acid sequence predicted from these DNAs and those of Japanese strains [To-336 vaccine strain (To-336 vac) and its wild progenitor (To-336 wt)]. The results suggest that the rubella viruses (RV) prevalent in certain areas of Japan in 2004 were highly homologous and were closely related with Japanese vaccine strain. PMID:16547415

  18. Crystal strength by direct computation

    NASA Astrophysics Data System (ADS)

    Bulatov, Vasily

    2007-03-01

    The art of making materials stronger goes back to medieval and even ancient times. Swords forged from Damascus steels more than 10 centuries ago possessed a unique combination of hardness and flexibility, two qualities that are difficult to attain simultaneously. The skills of metalworking were based on empirical knowledge and were passed from the master smith to his pupils. The science of physical metallurgy came about only in the XX century bringing with it new methods for finding out why some materials are strong while others are not. Soon it was realized that, when it comes to metal strength, it is all about crystal defects -- impurities, dislocations, grain boundaries, etc. - and how they are organized into crystal microstructure. This understanding has since resulted in new effective methods of material processing aiming to modify crystal microstructure in order to affect material's properties, e.g. strength and/or hardness. Remarkably and disappointingly, general understanding that microstructure defines material's response to external loads has not yet resulted in a workable physical theory of metal strength accounting for the realistic complexity of material microstructure. In this presentation I would like to discuss a few tidbits from computational and experimental research in our group at LLNL on crystal defects and their contributions to material strength. My selection of the examples aims to illustrate the major premise of our work that the mechanisms by which the microstructure affects crystal strength are multiple and complex but that there is hope to bring some order to this complexity.

  19. Calculation of Radiative Corrections to E1 matrix elements in the Neutral Alkalis

    SciTech Connect

    Sapirstein, J; Cheng, K T

    2004-09-28

    Radiative corrections to E1 matrix elements for ns-np transitions in the alkali metal atoms lithium through francium are evaluated. They are found to be small for the lighter alkalis but significantly larger for the heavier alkalis, and in the case of cesium much larger than the experimental accuracy. The relation of the matrix element calculation to a recent decay rate calculation for hydrogenic ions is discussed, and application of the method to parity nonconservation in cesium is described.

  20. Prenatal antidepressant exposure associated with CYP2E1 DNA methylation change in neonates

    PubMed Central

    Gurnot, Cécile; Martin-Subero, Ignacio; Mah, Sarah M; Weikum, Whitney; Goodman, Sarah J; Brain, Ursula; Werker, Janet F; Kobor, Michael S; Esteller, Manel; Oberlander, Tim F; Hensch, Takao K

    2015-01-01

    Some but not all neonates are affected by prenatal exposure to serotonin reuptake inhibitor antidepressants (SRI) and maternal mood disturbances. Distinguishing the impact of these 2 exposures is challenging and raises critical questions about whether pharmacological, genetic, or epigenetic factors can explain the spectrum of reported outcomes. Using unbiased DNA methylation array measurements followed by a detailed candidate gene approach, we examined whether prenatal SRI exposure was associated with neonatal DNA methylation changes and whether such changes were associated with differences in birth outcomes. Prenatal SRI exposure was first associated with increased DNA methylation status primarily at CYP2E1(βNon-exposed = 0.06, βSRI-exposed = 0.30, FDR = 0); however, this finding could not be distinguished from the potential impact of prenatal maternal depressed mood. Then, using pyrosequencing of CYP2E1 regulatory regions in an expanded cohort, higher DNA methylation status—both the mean across 16 CpG sites (P < 0.01) and at each specific CpG site (P < 0.05)—was associated with exposure to lower 3rd trimester maternal depressed mood symptoms only in the SRI-exposed neonates, indicating a maternal mood x SRI exposure interaction. In addition, higher DNA methylation levels at CpG2 (P = 0.04), CpG9 (P = 0.04) and CpG10 (P = 0.02), in the interrogated CYP2E1 region, were associated with increased birth weight independently of prenatal maternal mood, SRI drug exposure, or gestational age at birth. Prenatal SRI antidepressant exposure and maternal depressed mood were associated with altered neonatal CYP2E1 DNA methylation status, which, in turn, appeared to be associated with birth weight. PMID:25891251

  1. Aminotriazole Alleviates Acetaminophen Poisoning via Downregulating P450 2E1 and Suppressing Inflammation

    PubMed Central

    Ai, Qing; Ge, Pu; Dai, Jie; Jiang, Rong; Zhou, Dan; Che, Qian; Wan, Jingyuan; Zhang, Li

    2015-01-01

    Aminotriazole (ATZ) is commonly used as a catalase (CAT) inhibitor. We previously found ATZ attenuated oxidative liver injury, but the underlying mechanisms remain unknown. Acetaminophen (APAP) overdose frequently induces life-threatening oxidative hepatitis. In the present study, the potential hepatoprotective effects of ATZ on oxidative liver injury and the underlying mechanisms were further investigated in a mouse model with APAP poisoning. The experimental data indicated that pretreatment with ATZ dose- and time-dependently suppressed the elevation of plasma aminotransferases in APAP exposed mice, these effects were accompanied with alleviated histological abnormality and improved survival rate of APAP-challenged mice. In mice exposed to APAP, ATZ pretreatment decreased the CAT activities, hydrogen peroxide (H2O2) levels, malondialdehyde (MDA) contents, myeloperoxidase (MPO) levels in liver and reduced TNF-α levels in plasma. Pretreatment with ATZ also downregulated APAP-induced cytochrome P450 2E1 (CYP2E1) expression and JNK phosphorylation. In addition, posttreatment with ATZ after APAP challenge decreased the levels of plasma aminotransferases and increased the survival rate of experimental animals. Posttreatment with ATZ had no effects on CYP2E1 expression or JNK phosphorylation, but it significantly decreased the levels of plasma TNF-α. Our data indicated that the LD50 of ATZ in mice was 5367.4 mg/kg body weight, which is much higher than the therapeutic dose of ATZ in the present study. These data suggested that ATZ might be effective and safe in protect mice against APAP-induced hepatotoxicity, the beneficial effects might resulted from downregulation of CYP2E1 and inhibiton of inflammation. PMID:25884831

  2. Quadratic Hamilton-Poisson systems on $\\mathfrak{s}\\mathfrak{e}(1, 1)^{*}_{-}$: The homogeneous case

    NASA Astrophysics Data System (ADS)

    Barrett, Dennis I.; Biggs, Rory; Remsing, Claudiu C.

    2015-11-01

    In this paper we consider quadratic Hamilton-Poisson systems on the semi-Euclidean Lie-Poisson space {s}{e}(1, 1)*-. The homogeneous positive semidefinite systems are classified; there are exactly six equivalence classes. In each case, the stability nature of the equilibrium states is determined. Explicit expressions for the integral curves are found. A characterization of the equivalence classes, in terms of the equilibria, is identified. Finally, the relation of this work to optimal control is briefly discussed.

  3. 26 CFR 301.6223(e)-1 - Effect of Internal Revenue Service's failure to provide notice.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... years beginning prior to October 4, 2001, see § 301.6223(e)-1T contained in 26 CFR part 1, revised April...: Example 1. Partnership ABC has as one of its partners, A, a partnership with three partners, X, Y, and Z...). In addition, Z was identified as provided in section 6223(c)(3) and § 301.6223(c)-1 on May 1,...

  4. Ultrasound associated uptake of chitosan nanoparticles in MC3T3-E1 cells

    NASA Astrophysics Data System (ADS)

    Wu, Junyi

    Chitosan is a natural linear polysaccharide that has been well known for its applications in drug delivery system due to its unique physicochemical and biological properties. However, challenges still remain for it to become a fully realized therapeutic agent. In this study, we investigated the uptake of chitosan nanoparticles (CNP) under the ultrasound stimulation, using a model cell culture system (MC3T3-E1 mouse pre-osteoblasts). The CNP were fabricated by an ionic gelation method and were lyophilized prior to characterization and delivery to cells. Particle size and zeta potential were measured using Dynamic Light Scattering (DLS); the efficiency of chitosan complexation was measured using the ninhydrin assay. Cytotoxicity was examined by neutral red assay within 48 hours after delivery. The effect of ultrasound (US) on the efficiency of nanoparticle delivery to the MC3T3-E1 cells was examined at 1MHz and at either 1 or 2 W/cm2. Fluorescein isothiocyanate (FITC)-conjugated-CNP were used to visualize the internalized particles within the cytosol. The uptake of FITC-CNP exhibits a dose and time dependent effect, a strong FITC fluorescence was detected at the concentration of 500microg/mL under fluorescence microscope. Ultrasound assisted uptake of FITC-CNP performed a significant positive effect at 2W/cm2 with 60s of ultrasound exposure time. CNP displayed a slightly decrease in cell viability from 25microg/mL to 100microg/mL, while higher concentration of CNP facilitates the proliferation of MC3T3-E1 cells. Less than 10% of reduction in cell viability was observed for US at 1W/cm2 and 2W/cm2 with 30s and 60s of exposure time, which suggest a mild effect of US to MC3T3-E1 cell line.

  5. Mouse Spermatocytes Express CYP2E1 and Respond to Acrylamide Exposure

    PubMed Central

    Nixon, Belinda J.; Katen, Aimee L.; Stanger, Simone J.; Schjenken, John E.; Nixon, Brett; Roman, Shaun D.

    2014-01-01

    Metabolism of xenobiotics by cytochrome P450s (encoded by the CYP genes) often leads to bio-activation, producing reactive metabolites that interfere with cellular processes and cause DNA damage. In the testes, DNA damage induced by xenobiotics has been associated with impaired spermatogenesis and adverse effects on reproductive health. We previously reported that chronic exposure to the reproductive toxicant, acrylamide, produced high levels of DNA damage in spermatocytes of Swiss mice. CYP2E1 metabolises acrylamide to glycidamide, which, unlike acrylamide, readily forms adducts with DNA. Thus, to investigate the mechanisms of acrylamide toxicity in mouse male germ cells, we examined the expression of the CYP, CYP2E1, which metabolises acrylamide. Using Q-PCR and immunohistochemistry, we establish that CYP2E1 is expressed in germ cells, in particular in spermatocytes. Additionally, CYP2E1 gene expression was upregulated in these cells following in vitro acrylamide exposure (1 µM, 18 h). Spermatocytes were isolated and treated with 1 µM acrylamide or 0.5 µM glycidamide for 18 hours and the presence of DNA-adducts was investigated using the comet assay, modified to detect DNA-adducts. Both compounds produced significant levels of DNA damage in spermatocytes, with a greater response observed following glycidamide exposure. A modified comet assay indicated that direct adduction of DNA by glycidamide was a major source of DNA damage. Oxidative stress played a small role in eliciting this damage, as a relatively modest effect was found in a comet assay modified to detect oxidative adducts following glycidamide exposure, and glutathione levels remained unchanged following treatment with either compound. Our results indicate that the male germ line has the capacity to respond to xenobiotic exposure by inducing detoxifying enzymes, and the DNA damage elicited by acrylamide in male germ cells is likely due to the formation of glycidamide adducts. PMID:24788432

  6. Prenatal antidepressant exposure associated with CYP2E1 DNA methylation change in neonates.

    PubMed

    Gurnot, Cécile; Martin-Subero, Ignacio; Mah, Sarah M; Weikum, Whitney; Goodman, Sarah J; Brain, Ursula; Werker, Janet F; Kobor, Michael S; Esteller, Manel; Oberlander, Tim F; Hensch, Takao K

    2015-01-01

    Some but not all neonates are affected by prenatal exposure to serotonin reuptake inhibitor antidepressants (SRI) and maternal mood disturbances. Distinguishing the impact of these 2 exposures is challenging and raises critical questions about whether pharmacological, genetic, or epigenetic factors can explain the spectrum of reported outcomes. Using unbiased DNA methylation array measurements followed by a detailed candidate gene approach, we examined whether prenatal SRI exposure was associated with neonatal DNA methylation changes and whether such changes were associated with differences in birth outcomes. Prenatal SRI exposure was first associated with increased DNA methylation status primarily at CYP2E1(β(Non-exposed) = 0.06, β(SRI-exposed) = 0.30, FDR = 0); however, this finding could not be distinguished from the potential impact of prenatal maternal depressed mood. Then, using pyrosequencing of CYP2E1 regulatory regions in an expanded cohort, higher DNA methylation status--both the mean across 16 CpG sites (P < 0.01) and at each specific CpG site (P < 0.05)--was associated with exposure to lower 3rd trimester maternal depressed mood symptoms only in the SRI-exposed neonates, indicating a maternal mood x SRI exposure interaction. In addition, higher DNA methylation levels at CpG2 (P = 0.04), CpG9 (P = 0.04) and CpG10 (P = 0.02), in the interrogated CYP2E1 region, were associated with increased birth weight independently of prenatal maternal mood, SRI drug exposure, or gestational age at birth. Prenatal SRI antidepressant exposure and maternal depressed mood were associated with altered neonatal CYP2E1 DNA methylation status, which, in turn, appeared to be associated with birth weight. PMID:25891251

  7. Resveratrol Pretreatment Affects CYP2E1 Activity of Chlorzoxazone in Healthy Human Volunteers.

    PubMed

    Bedada, Satish Kumar; Neerati, Prasad

    2016-03-01

    The purpose of the present study was to investigate the effect of resveratrol (RSV) pretreatment on CYP2E1 enzyme activity and pharmacokinetics of chlorzoxazone (CHZ) in healthy human volunteers. The open-label, two period, sequential study was conducted in 12 healthy human volunteers. A single dose of RSV 500?mg was administered once daily for 10?days during treatment phase. A single dose of CHZ 250?mg was administered during control and after treatment phases under fasting conditions. The blood samples were collected after CHZ dosing at predetermined time intervals and analyzed by HPLC. RSV pretreatment significantly enhanced the maximum plasma concentration (Cmax ), area under the curve (AUC) and half life (T1/2 ) and significantly decreased elimination rate constant (Kel ), apparent oral clearance (CL/F) and apparent volume of distribution (Vd/F) of CHZ as compared to that of control. In addition, RSV pretreatment significantly decreased the metabolite to parent (6-OHCHZ/CHZ) ratios of Cmax , AUC and T1/2 and significantly increased the Kel ratio of 6-OHCHZ/CHZ, which indicated the reduced formation of CHZ to 6-OHCHZ. The results suggest that the altered CYP2E1 enzyme activity and pharmacokinetics of CHZ might be attributed to RSV mediated inhibition of CYP2E1 enzyme. Thus, there is a potential pharmacokinetic interaction between RSV and CHZ. The inhibition of CYP2E1 by RSV may provide a novel approach for minimizing the hepatotoxicity of ethanol. Copyright 2015 John Wiley & Sons, Ltd. PMID:26680654

  8. Fluxes and spectra of quasimonochromatic annihilation photons for studying E1 giant resonances in nuclei

    SciTech Connect

    Dzhilavyan, L. Z.

    2014-12-15

    The fluxes and spectra of quasimonochromatic photons originating from the in-flight annihilation of positrons interacting with electrons of targets are analyzed in the energy region characteristic of the excitation of E1 giant resonances in nuclei. Targets of small thickness and low atomic number are used. The dependences of the spectra on the energy and angle (and their scatter) for positrons incident to the target, on the collimation angle for photons, and on the target thickness are studied.

  9. The possible mechanisms of CYP2E1 interactions with HSP90 and the influence of ethanol on them

    PubMed Central

    2012-01-01

    Background Microsomal CYP2E1 metabolizes about 160 hydrophobic exogens, many of which are environmental pollutants. While metabolising xenobiotics CYP2E1 on one hand facilitates in their excretion and on the other hand activates them into the cytotoxins, which may damage the cell. Thus the CYP2E1 activity level significantly affects the processes in cell. Posttranslational stabilization of CYP2E1 seems to be the main mechanism of its regulation in living cell. It is known that degradation of CYP2El takes part in cytoplasmic proteasome system. The efficiency of such degradation depends on the presence of molecular chaperones (HSP90) as was shown from in vitro experiments. But the processes that involve HSP90 in the degradation of CYP2E1 and the mechanisms of transfer of microsomal CYP2E1 to the proteasome system remain unknown. This paper investigates HSP90-dependent processes in mechanisms of CYP2El degradation and the possible role of ethanol in them. Results With the help of computational methods we have shown that CYP2E1 can interact with HSP90 resulting in dissociation of CYP2E1 from membrane and formation of the CYP2E1-HSP90 complex for its further transfer to the proteasome for degradation. The twofold increase of both CYP2E1 and HSP90 in the mouse liver under the constant alcohol administration was shown using WB methods. Also, as was shown in silico, ethanol molecule, while binding to the CYP2E1 active site, prevents its interaction with HSP90, thus resulting in accumulation of CYP2E1 in cell. Conclusions Cytoplasmic HSP90 and membrane-bound CYP2E1 may directly interact with each other as partner proteins, leading to the dissociation of the CYP2E1 from the membrane. This makes it possible to transfer microsomal CYP2E1 in complex with HSP90 to the proteasome for proteolysis. The ethanol molecule inhibits the interaction of HSP90 with CYP2E1 leading to the suppression of its proteasome degradation, thus increasing level of this protein in the cell. Other substrates of CYP2E1 should increase level of this protein in the same way. This may be one of the mechanisms of substrate-dependent regulation of the CYP2E1 expression in the cell. PMID:23241420

  10. Precision measurement of the electromagnetic dipole strengths in Be11

    NASA Astrophysics Data System (ADS)

    Kwan, E.; Wu, C. Y.; Summers, N. C.; Hackman, G.; Drake, T. E.; Andreoiu, C.; Ashley, R.; Ball, G. C.; Bender, P. C.; Boston, A. J.; Boston, H. C.; Chester, A.; Close, A.; Cline, D.; Cross, D. S.; Dunlop, R.; Finlay, A.; Garnsworthy, A. B.; Hayes, A. B.; Laffoley, A. T.; Nano, T.; Navrátil, P.; Pearson, C. J.; Pore, J.; Quaglioni, S.; Svensson, C. E.; Starosta, K.; Thompson, I. J.; Voss, P.; Williams, S. J.; Wang, Z. M.

    2014-05-01

    The electromagnetic dipole strength in Be11 between the bound states has been measured using low-energy projectile Coulomb excitation at bombarding energies of 1.73 and 2.09 MeV/nucleon on a Pt196 target. An electric dipole transition probability B(E1;1/2-→1/2+)=0.102(2) e2fm was determined using the semi-classical code Gosia, and a value of 0.098(4) e2fm was determined using the Extended Continuum Discretized Coupled Channels method with the quantum mechanical code FRESCO. These extracted B(E1) values are consistent with the average value determined by a model-dependent analysis of intermediate energy Coulomb excitation measurements and are approximately 14% lower than that determined by a lifetime measurement. The much-improved precisions of 2% and 4% in the measured B(E1) values between the bound states deduced using Gosia and the Extended Continuum Discretized Coupled Channels method, respectively, compared to the previous accuracy of ˜10% will help in our understanding of and better improve the realistic inter-nucleon interactions.

  11. Instanton induced Yukawa couplings from distant E3 and E(-1) instantons

    NASA Astrophysics Data System (ADS)

    Goodsell, Mark D.; Witkowski, Lukas T.

    2016-01-01

    We calculate non-perturbative contributions to Yukawa couplings on D3-branes at orbifold singularities due to E3 and fractional E(-1) instantons which do not intersect the visible sector branes. While distant E3 instantons on bulk cycles typically contribute to Yukawa couplings, we find that distant fractional E(-1) can also give rise to new Yukawa couplings. However, fractional E(-1) instantons only induce Yukawa couplings if they are located at a singularity which shares a collapsed homologous two-cycle with the singularity supporting the visible sector. The non-perturbative contributions to Yukawa couplings exhibit a different flavour structure than the tree-level Yukawa couplings and, as a result, they can be sources of flavour violation. This is particularly relevant for schemes of moduli stabilisation which rely on superpotential contributions from E3 instantons, such as KKLT or the Large Volume Scenario. As a byproduct of our analysis, we shed some new light on the properties of annulus diagrams with matter field insertions in stringy instanton calculus.

  12. A comparison of substrate dynamics in human CYP2E1 and CYP2A6

    PubMed Central

    Harrelson, John P.; Henne, Kirk R.; Alonso, Darwin O.V.; Nelson, Sidney D.

    2009-01-01

    Considering the dynamic nature of CYPs, methods that reveal information about substrate and enzyme dynamics are necessary to generate predictive models. To compare substrate dynamics in CYP2E1 and CYP2A6, intramolecular isotope effect experiments were conducted, using deuterium labeled substrates: o-xylene, m-xylene, p-xylene, 2,6-dimethylnaphthalene, and 4,4?-dimethylbiphenyl. Competitive intermolecular experiments were also conducted using d0- and d6-labeled p-xylene. Both CYP2E1 and CYP2A6 displayed full isotope effect expression for o-xylene oxidation and almost complete suppression for dimethylbiphenyl. Interestingly, (kH/kD)obs for d3-p-xylene oxidation ((kH/kD)obs = 6.04 and (kH/kD)obs = 5.53 for CYP2E1 and CYP2A6, respectively) was only slightly higher than (kH/kD)obs for d3-dimethylnaphthalene ((kH/kD)obs = 5.50 and (kH/kD)obs = 4.96, respectively). One explanation is that in some instances (kH/kD)obs values are generated by the presence of two substrates bound simultaneously to the CYP. Speculatively, if this explanation is valid, then intramolecular isotope effect experiments should be useful in the mechanistic investigation of P450 cooperativity. PMID:17156750

  13. Contribution of CYP2E1 polymorphism to aging in the mechanical workshop workers.

    PubMed

    Eshkoor, Sima Ataollahi; Ismail, Patimah; Rahman, Sabariah Abd; Adon, Mohd Yusoff; Devan, R Vasu

    2013-05-01

    Aging is attributed to both genetic and environmental factors. Occupational exposure is one of the environmental factors with potential genotoxic effects. Researchers try to determine factors involved in genetic damages at hazards exposure that could accelerate aging. Cytochrome P450 2E1 (CYP2E1) gene contributes in activation and detoxification of the environmental hazards. This polymorphism plays an important role in susceptibility of inter-individuals to DNA damage at the occupational exposure. The current study evaluated the possible influence of this gene polymorphism in aging by genomic damages through the biomarkers alterations of micronuclei (MN), comet tail length and telomere length shortening at the exposure. In this study, buccal cells were collected from the oral cavity of exposed workers and non-exposed controls. The CYP2E1 genotypes were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The wild genotype significantly affected MN frequency (p = 0.007) and relative telomere length (p = 0.047) in the older group of workers. It was concluded that the interaction of gene polymorphism and exposure enhances DNA damage and accelerates aging consequently. PMID:23193996

  14. Bursts of energetic electron induced large surface charging observed by Chang'E-1

    NASA Astrophysics Data System (ADS)

    Wang, X. Y.; Zhang, A. B.; Zhang, X. G.; Reme, H.; Kong, L. G.; Zhang, S. Y.; Yu, D. J.; Wang, S. J.; Zhu, G. W.

    2012-10-01

    A relationship between surface charging and bursts of energetic electron (BEE) event is presented in this paper. In a 200 km lunar polar orbit, during quiet time, 0.1-2.0 MeV BEE events were observed by High Energetic Particles Detectors (HPD) on board Chang'E-1, on December 22, 2007, when the spacecraft was within the inner terrestrial magnetosheath. At the same time, a large surface charging of ∼-5.4 kV was observed by Chang'E-1, which was evidenced by increasing the ions energy observed by Solar Wind Ion Detectors (SWIDs). We found that the surface charging is strongly correlated with BEE events, and the potentials of spacecraft surface charging was experientially expressed as U≈3.6×10-5·fT (kV). The BEE events did occur in the solar wind, geomagnetic tail and magnetosheath alternately, whereas the surface charging during the BEE events is in the magnetosheath or transition region of boundaries. Though the observed surface charging was fewer than the BEE events, it is expected that the occurrence of the charging events caused by the bursts of energetic electrons should be more frequent than the Chang'E-1 observations. Meanwhile, the spacecraft charging indicates the lunar surface can be charged to negative kilovolt-scale by the BEE events even in quiet times.

  15. Role of an adenovirus E2 promoter binding factor in E1A-mediated coordinate gene control.

    PubMed Central

    Kovesdi, I; Reichel, R; Nevins, J R

    1987-01-01

    A product of the adenovirus gene E1A is responsible for the stimulation of transcription from six viral promoters as well as at least two cellular promoters. We have detected a HeLa cell factor, termed E2 promoter binding factor (E2F), that appears to mediate the transcriptional stimulation of the viral E2 promoter. Competition experiments revealed that E2F did not recognize and bind to the E1B, E3, E4, or major late promoter sequences. Furthermore, three additional promoters stimulated by E1A, heat shock protein 70, beta-globin, and early simian virus 40, do not bind E2F. In contrast, the factor does recognize sequences in the E1A enhancer, and within the E1A enhancer are duplicated binding sites for E2F. Finally, a single E2F binding site from the E1A enhancer can confer increased transcription to a mouse beta-globin promoter, dependent on the action of the E1A gene product. This stimulation requires binding of E2F since methylation of the binding site, which blocks binding in vitro, reduces transcription stimulation in vivo. We, therefore, conclude that E2F is likely to be responsible for the E1A-mediated stimulation of the E1A gene as well as the E2 gene but is not involved in the activation of the other E1A-inducible promoters. Images PMID:2951737

  16. Polymorphisms in the CYP2E1 and GSTM1 Genes as Possible Protection Factors for Leprosy Patients

    PubMed Central

    Pinto, Pablo; Salgado, Claudio Guedes; Santos, Ney; Alencar, Dayse O.; Santos, Sidney; Hutz, Mara H.; Ribeiro-dos-Santos, Ândrea

    2012-01-01

    Background The CYP2E1 and GSTM1 genes encode metabolic enzymes that have key functions in drug modification and elimination. Methodology/Principal Findings We investigated the possible effects of CYP2E1 and GSTM1 polymorphisms in 71 leprosy patients and in 110 individuals from the general population. The GSTM1*0 null allele and INDEL CYP2E1*1D mutant genotypes were analyzed by conventional PCR, while CYP2E1 SNPs (1053C>T, 1293G>C and 7632T>A) were determined by RT-PCR. In leprosy patients, the GSTM1*0 and CYP2E1*5 alleles and the combined alleles GSTM1*0/CYP2E1*6 and GSTM1*0/CYP2E1*5 were significantly related to a baciloscopic index (BI) (BI<3), while the CYP2E1*6 allele was related to a better clinical evolution in the leprosy spectrum. Conclusions/Significance Therefore, GSTM1*0, CYP2E1*5 and CYP2E1*6 may be possible protection factors for leprosy patients. PMID:23077626

  17. Chimeric Derivatives of Hepatitis B Virus Core Particles Carrying Major Epitopes of the Rubella Virus E1 Glycoprotein

    PubMed Central

    Skrastina, Dace; Petrovskis, Ivars; Petraityte, Rasa; Sominskaya, Irina; Ose, Velta; Liekniņa, Ilva; Bogans, Janis; Sasnauskas, Kestutis

    2013-01-01

    Three variants of the major rubella virus (RV) E1 protein virus-neutralizing epitope from position 214 to 285 were exposed on the hepatitis B virus (HBV) C-terminally truncated core (HBcΔ) in a virus-like particle (VLP) vector and were produced in Escherichia coli. All three chimeras demonstrated VLPs in bacterial cell lysates, but only HBcΔ-E1(245-285) demonstrated the correct VLP structure after purification. The other chimeras, HBcΔ-E1(214-285) and HBcΔ-E1(214-240), appeared after purification as non-VLP aggregates of 100 to 900 nm in diameter according to dynamic light scattering data. All three variants possessed the intrinsic antigenic activity of RV E1, since they were recognized by natural human anti-RV E1 antibodies and induced an anti-RV E1 response in mice. HBcΔ-E1(214-240) and HBcΔ-E1(245-285) can be regarded as prototypes for a putative RV vaccine because they were able to induce antibodies recognizing natural RV E1 protein in RV diagnostic kits. PMID:24006140

  18. Strength of Nanotubes and Megacables

    NASA Astrophysics Data System (ADS)

    Pugno, Nicola M.

    In this chapter my findings [mainly reported in N. Pugno, J. Phys.- Condens. Matter, 18, S1971-S1990 (2006); N. Pugno, Acta Mater. 55, 5269-5279 (2007); N. Pugno, Nano Today 2, 44-47 (2007)] on the mechanical strength of nanotubes and megacables are reviewed, with an eye to the challenging project of the carbon nanotube-based space elevator megacable. Accordingly, basing the design of the megacable on the theoretical strength of a single carbon nanotube, as originally proposed at the beginning of the third millennium, has been demonstrated to be naïve. The role on the fracture strength of thermodynamically unavoidable atomistic defects with different size and shape is thus here quantified on brittle fracture both numerically (with ad hoc hierarchical simulations) and theoretically (with quantized fracture theories), for nanotubes and nanotube bundles. Fatigue, elasticity, non-asymptotic regimes, elastic-plasticity, rough cracks, finite domains and size-effects are also discussed.

  19. Cytochrome P450 2E1 genetic polymorphism and gastric cancer in Changle, Fujian Province

    PubMed Central

    Cai, Lin; Yu, Shun-Zhang; Zhang, Zuo-Feng

    2001-01-01

    AIM: Genetic polymorphism in enzymes of carcinogen metabolism has been found to have the influence on the susceptibility to cancer. Cytochrome P450 2E1 (CYP2E1) is considered to play an important role in the metabolic activation of procarcinogens such as N-nitrosoamines and low molecular weight organic compounds. The purpose of this study is to determine whether CYP450 2E1 polymorphisms are associated with risks of gastric cancer. METHODS: We conducted a population based case-control study in Changle county, Fujian Province, a high-risk region of gastric cancer in China. Ninety-one incident gastric cancer patients and ninety-four healthy controls were included in our study. Datas including demographic characteristcs, diet intake, and alcohol and tobacco consumption of indivduals in our study were completed by a standardized questionnaire. PCR-RFLP revealed three genotypes:heterozygote (C1/C2) and two homozygotes (C1/C1 and C2/C2) in CYP2E1. RESULTS: The frequency of variant genotypes (C1/C2 and C2/C2) in gastric cancer cases and controls was 36.3% and 24.5%, respectively. The rare homozygous C2/C2 genotype was found in 6 indivduals in gastric cancer group (6.6%), whereas there was only one in the control group (1.1%). However, there was no statistically significan difference between the two groups (two-tailed Fisher’s exact test, P = 0.066). Indivduals in gastric cancer group were more likely to carry genotype C1/C2 (odds ratio, OR = 1.50) and C2/C2 (OR = 7.34) than indivduals in control group (χ² = 4.597, for trend P = 0.032). The frequencies of genotypes with the C2 allele (C1/C2 and C2/C2 genotypes) were compared with those of genotypes without C2 allele (C1/C1 genotype) among indivduals in gastric cancer group and control group according to the pattern of gastric cancer risk factors. The results show that indivduals who exposed to these gastric cancer risk factors and carry the C2 allele seemed to have a higher risk of developing gastric cancer. CONCLUSION: Polymorphism of CYP2E1 gene may have some effct in the development of gastric cancer in Changle county, Fujian Province. PMID:11854903

  20. Cytochrome P450 2E1 RsaI/PstI polymorphism is associated with urologic cancer risk: evidence from a meta-analysis

    PubMed Central

    Lin, You-Cheng; Wu, Xun; Zhou, Xue-Qiong; Ren, Rui; Su, Ze-Xuan; Liu, Chun-Xiao

    2015-01-01

    Cytochrome P450 2E1 (CYP2E1) is involved in the metabolic activation of various carcinogens. CYP2E1 RsaI/PstI polymorphism has been identified in urologic cancer patients, while studies of the polymorphism have shown inconclusive trends in the risk of urologic cancers. Therefore, we performed this systematic review to provide a complete picture and conducted a meta-analysis to derive a precise estimation. We searched PubMed, Embase and Web of Science to identify eligible studies up to December 15, 2014. 12 studies with 2712 cases and 2977 controls were included in the meta-analysis.The odds ratio with a 95% confidence interval was used to assess the strength of associations. We observed that the c2 allele of CYP2E1 RsaI/PstI polymorphism was associated with a decreased risk of urologic cancer under all genetic models (c2 vs. c1: OR = 0.742, 95% CI = 0.659-0.835); c2c2 vs. c1c1: OR = 0.516, 95% CI = 0.357-0.745; c1c2 vs. c1c1: OR = 0.748, 95% CI = 0.748 (0.648-0.863; c2c2 + c1c2 vs. c1c1: OR = 0.722, 95% CI = 0.629-0.829; c2c2 vs. c1c1 + c1c2: OR = 0.578, 95% CI = 0.401-0.832). In the subgroup analysis by cancer type, statistically significant associations were found in urothelial cancer in all genetic models. When stratified by ethnicity, a same trend was also indicated in Asians in all genetic models.To conclude, our results support the conclusion that the CYP2E1 RsaI/PstI polymorphism may be associated with urologic cancer susceptibility. The c2 allele is a low-penetrance risk factor for urologic cancer development. PMID:26309545

  1. Cytochrome P450 2E1 RsaI/PstI polymorphism is associated with urologic cancer risk: evidence from a meta-analysis.

    PubMed

    Lin, You-Cheng; Wu, Xun; Zhou, Xue-Qiong; Ren, Rui; Su, Ze-Xuan; Liu, Chun-Xiao

    2015-01-01

    Cytochrome P450 2E1 (CYP2E1) is involved in the metabolic activation of various carcinogens. CYP2E1 RsaI/PstI polymorphism has been identified in urologic cancer patients, while studies of the polymorphism have shown inconclusive trends in the risk of urologic cancers. Therefore, we performed this systematic review to provide a complete picture and conducted a meta-analysis to derive a precise estimation. We searched PubMed, Embase and Web of Science to identify eligible studies up to December 15, 2014. 12 studies with 2712 cases and 2977 controls were included in the meta-analysis.The odds ratio with a 95% confidence interval was used to assess the strength of associations. We observed that the c2 allele of CYP2E1 RsaI/PstI polymorphism was associated with a decreased risk of urologic cancer under all genetic models (c2 vs. c1: OR = 0.742, 95% CI = 0.659-0.835); c2c2 vs. c1c1: OR = 0.516, 95% CI = 0.357-0.745; c1c2 vs. c1c1: OR = 0.748, 95% CI = 0.748 (0.648-0.863; c2c2 + c1c2 vs. c1c1: OR = 0.722, 95% CI = 0.629-0.829; c2c2 vs. c1c1 + c1c2: OR = 0.578, 95% CI = 0.401-0.832). In the subgroup analysis by cancer type, statistically significant associations were found in urothelial cancer in all genetic models. When stratified by ethnicity, a same trend was also indicated in Asians in all genetic models.To conclude, our results support the conclusion that the CYP2E1 RsaI/PstI polymorphism may be associated with urologic cancer susceptibility. The c2 allele is a low-penetrance risk factor for urologic cancer development. PMID:26309545

  2. CYP2E1-catalyzed alcohol metabolism: role of oxidant generation in interferon signaling, antigen presentation and autophagy.

    PubMed

    Osna, Natalia A; Donohue, Terrence M

    2013-01-01

    Cytochrome P450 2E1 (CYP2E1) is one of two major enzymes that catalyze ethanol oxidation in the liver. CYP2E1 is also unique because it is inducible, as its hepatic content rises after continuous (chronic) ethanol administration, thereby accelerating the rate of ethanol metabolism and affording greater tolerance to heavy alcohol consumption. However, the broad substrate specificity of CYP2E1 and its capacity to generate free radicals from alcohol and other hepatotoxins, places CYP2E1 as a central focus of not only liver toxicity, but also as an enzyme that regulates cytokine signaling, antigen presentation, and macromolecular degradation, all of which are crucial to liver cell function and viability. Here, we describe our own and other published work relevant to the importance of CYP2E1-catalyzed ethanol oxidation and how this catalysis affects the aforementioned cellular processes to produce liver injury. PMID:23400922

  3. Mitochondria-targeted cytochrome P450 2E1 induces oxidative damage and augments alcohol-mediated oxidative stress.

    PubMed

    Bansal, Seema; Liu, Chuan-Peng; Sepuri, Naresh B V; Anandatheerthavarada, Hindupur K; Selvaraj, Venkatesh; Hoek, Jan; Milne, Ginger L; Guengerich, F Peter; Avadhani, Narayan G

    2010-08-01

    The ethanol-inducible cytochrome P450 2E1 (CYP2E1) is also induced under different pathological and physiological conditions. Studies including ours have shown that CYP2E1 is bimodally targeted to both the endoplasmic reticulum (microsomes) (mc CYP2E1) and mitochondria (mt CYP2E1). In this study we investigated the role of mtCYP2E1 in ethanol-mediated oxidative stress in stable cell lines expressing predominantly mt CYP2E1 or mc CYP2E1. The ER+ mutation (A2L, A9L), which increases the affinity of the nascent protein for binding to the signal recognition particle, preferentially targets CYP2E1 to the endoplasmic reticulum. The Mt+ (L17G) and Mt++ (I8R, L11R, L17R) mutant proteins, showing progressively lower affinity for signal recognition particle binding, were targeted to mitochondria at correspondingly higher levels. The rate of GSH depletion, used as a measure of oxidative stress, was higher in cells expressing Mt++ and Mt+ proteins as compared with cells expressing ER+ protein. In addition, the cellular level of F(2)-isoprostanes, a direct indicator of oxidative stress, was increased markedly in Mt++ cells after ethanol treatment. Notably, expression of Mt++ CYP2E1 protein in yeast cells caused more severe mitochondrial DNA damage and respiratory deficiency than the wild type or ER+ proteins as tested by the inability of cells to grow on glycerol or ethanol. Additionally, liver mitochondria from ethanol-fed rats containing high mt CYP2E1 showed higher levels of F(2)-isoprostane production. These results strongly suggest that mt CYP2E1 induces oxidative stress and augments alcohol-mediated cell/tissue injury. PMID:20529841

  4. Prepubescent Strength Training Gains Support.

    ERIC Educational Resources Information Center

    Duda, Marty

    1986-01-01

    Recent studies have stimulated greater support for prepubescent weight training. There seems to be general agreement that strength and weight training, when practiced under properly controlled conditions, is safe and efficacious for prepubescents. Weight lifting is not supported. Recommendations for weight training are made, and reservations are…

  5. The Strengths of Black Families.

    ERIC Educational Resources Information Center

    Hill, Robert B.

    This report identifies and analyzes five strengths of black families: adaptability roles, strong kinship bonds, strong work orientation, strong religious orientation, and achievement orientation. These five characteristics have been functional for the survival, advancement, and stability of black families. Most discussions of black families tend…

  6. Prepubescent Strength Training Gains Support.

    ERIC Educational Resources Information Center

    Duda, Marty

    1986-01-01

    Recent studies have stimulated greater support for prepubescent weight training. There seems to be general agreement that strength and weight training, when practiced under properly controlled conditions, is safe and efficacious for prepubescents. Weight lifting is not supported. Recommendations for weight training are made, and reservations are

  7. Building Strengths through Adventure Education

    ERIC Educational Resources Information Center

    Loughmiller, Grover C.

    2007-01-01

    Campbell Loughmiller (1906-1993) is widely recognized as a leader in therapeutic work with troubled youngsters in outdoor settings. Rejecting punitive or institutional models of intervention, Loughmiller set out to demonstrate that every young person has strengths, desires to make positive changes, can grow in responsibility, and contribute to

  8. Characterization of novel cytochrome P450 2E1 knockout rat model generated by CRISPR/Cas9.

    PubMed

    Wang, Xin; Tang, Yu; Lu, Jian; Shao, Yanjiao; Qin, Xuan; Li, Yongmei; Wang, Liren; Li, Dali; Liu, Mingyao

    2016-04-01

    A bacterial CRISPR-associated protein-9 nuclease (CRISPR/Cas9) from Streptococcus pyogenes has generated considerable excitement as a new tool to edit the targeted genome. Cytochrome P450 (CYP) 2E1 not only plays an important role in the xenobiotic metabolism and chemical toxicity, but also is involved in many kinds of diseases, such as alcoholic liver diseases and diabetes. Despite its importance, few animal models are used to predict CYP2E1 properties in physiology, pathology, as well as carcinogen activation. To establish a novel model for investigating the functions of CYP2E1 in vivo, this study has successfully generated the Cyp2e1 knockout (KO) rat model without detectable off-target effects using CRISPR/Cas9 system. The Cyp2e1 KO rats were viable and fertile and did not display any obvious physiological abnormities. The absent expression of CYP2E1 in KO rats also resulted in inactive behaviors in the metabolism of CYP2E1 substrates. The Cyp2e1 KO rats as a novel and available rodent animal model provide a powerful tool for the study of CYP2E1 in the chemical metabolism, toxicity, carcinogenicity, and its core factor in drug-drug interactions. PMID:26947455

  9. Ethanol Induction of CYP2A5: Role of CYP2E1-ROS-Nrf2 Pathway

    PubMed Central

    Lu, Yongke; Zhang, Xu Hannah

    2012-01-01

    Chronic ethanol consumption was previously shown to induce CYP2A5 in mice, and this induction of CYP2A5 by ethanol was CYP2E1 dependent. In this study, the mechanisms of CYP2E1-dependent ethanol induction of CYP2A5 were investigated. CYP2E1 was induced by chronic ethanol consumption to the same degree in wild-type (WT) mice and CYP2A5 knockout (Cyp2a5 –/–) mice, suggesting that unlike the CYP2E1-dependent ethanol induction of CYP2A5, ethanol induction of CYP2E1 is not CYP2A5 dependent. Microsomal ethanol oxidation was about 25% lower in Cyp2a5 –/– mice compared with that in WT mice, suggesting that CYP2A5 can oxidize ethanol although to a lesser extent than CYP2E1 does. CYP2A5 was induced by short-term ethanol consumption in human CYP2E1 transgenic knockin (Cyp2e1 –/– KI) mice but not in CYP2E1 knockout (Cyp2e1 –/–) mice. The redox-sensitive transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) was also induced by acute ethanol in Cyp2e1 –/– KI mice but not in Cyp2e1 –/– mice. Ethanol induction of CYP2A5 in Nrf2 knockout (Nrf2 –/–) mice was lower compared with that in WT mice, whereas CYP2E1 induction by ethanol was comparable in WT and Nrf2 –/– mice. Antioxidants (N-acetyl-cysteine and vitamin C), which blocked oxidative stress induced by chronic ethanol in WT mice and acute ethanol in Cyp2e1 –/– KI mice, also blunted the induction of CYP2A5 and Nrf2 by ethanol but not the induction of CYP2E1 by ethanol. These results suggest that oxidative stress induced by ethanol via induction of CYP2E1 upregulates Nrf2 activity, which in turn regulates ethanol induction of CYP2A5. Results obtained from primary hepatocytes, mice gavaged with binge ethanol or fed chronic ethanol, show that Nrf2-regulated ethanol induction of CYP2A5 protects against ethanol-induced steatosis. PMID:22552773

  10. Characterization of papillomavirus E1 helicase mutants defective for interaction with the SUMO-conjugating enzyme Ubc9

    SciTech Connect

    Fradet-Turcotte, Amelie; Brault, Karine; Titolo, Steve; Howley, Peter M.; Archambault, Jacques

    2009-12-20

    The E1 helicase from BPV and HPV16 interacts with Ubc9 to facilitate viral genome replication. We report that HPV11 E1 also interacts with Ubc9 in vitro and in the yeast two-hybrid system. Residues in E1 involved in oligomerization (353-435) were sufficient for binding to Ubc9 in vitro, but the origin-binding and ATPase domains were additionally required in yeast. Nuclear accumulation of BPV E1 was shown previously to depend on its interaction with Ubc9 and sumoylation on lysine 514. In contrast, HPV11 and HPV16 E1 mutants defective for Ubc9 binding remained nuclear even when the SUMO pathway was inhibited. Furthermore, we found that K514 in BPV E1 and the analogous K559 in HPV11 E1 are not essential for nuclear accumulation of E1. These results suggest that the interaction of E1 with Ubc9 is not essential for its nuclear accumulation but, rather, depends on its oligomerization and binding to DNA and ATP.

  11. PP2A-B55? Antagonizes Cyclin E1 Proteolysis and Promotes its Dysregulation in Cancer

    PubMed Central

    Tan, YingMeei; Sun, Dahui; Jiang, Weijian; Klotz-Noack, Kathleen; Vashisht, Ajay A.; Wohlschlegel, James; Widschwendter, Martin; Spruck, Charles

    2014-01-01

    Cyclin E1 regulates the initiation of S phase in cellular division. However, in many cancers cyclin E1 is aberrantly overexpressed and this molecular phenotype correlates with increased tumor aggressiveness and poor patient survival. The molecular cause(s) of cyclin E1 abnormalities in cancers is poorly understood. Here, we show cyclin E1 overexpression in cancer is promoted by dysregulation of the protein phosphatase PP2A-B55?. PP2A-B55? targets the N- and C-terminal phosphodegrons of cyclin E1 for dephosphorylation, thus protecting it from degradation mediated by the SCFFbxw7 ubiquitin ligase. Augmented B55? expression stabilizes cyclin E1 and promotes its overexpression in cancer-derived cell lines and breast tumors. Conversely, B55? ablation enforces the degradation of cyclin E1 and inhibits cancer cell proliferation in vitro and tumor formation in vivo. Therefore, PP2A-B55? promotes cyclin E1 overexpression by antagonizing its degradation and its inhibition could represent a therapeutic mechanism for abrogating cyclin E1 function in cancers. PMID:24509904

  12. Flow perfusion culture of MC3T3-E1 osteogenic cells on gradient calcium polyphosphate scaffolds with different pore sizes.

    PubMed

    Chen, Liang; Song, Wei; Markel, David C; Shi, Tong; Muzik, Otto; Matthew, Howard; Ren, Weiping

    2016-02-01

    Calcium polyphosphate is a biodegradable bone substitute. It remains a challenge to prepare porous calcium polyphosphate with desired gradient porous structures. In this study, a modified one-step gravity sintering method was used to prepare calcium polyphosphate scaffolds with desired-gradient-pore-size distribution. The differences of porous structure, mechanical strength, and degradation rate between gradient and homogenous calcium polyphosphate scaffolds were evaluated by micro-computed tomography, scanning electron microscopy, and mechanical testing. Preosteoblastic MC3T3-E1 cells were seeded onto gradient and homogenous calcium polyphosphate scaffolds and cultured in a flow perfusion bioreactor. The distribution, proliferation, and differentiation of the MC3T3-E1 cells were compared to that of homogenous calcium polyphosphate scaffolds. Though no significant difference of cell proliferation was found between the gradient and the homogenous calcium polyphosphate scaffolds, a much higher cell differentiation and mineralization were observed in the gradient calcium polyphosphate scaffolds than that of the homogenous calcium polyphosphate scaffolds, as manifested by increased alkaline phosphatase activity (p < 0.05). The improved distribution and differentiation of cultured cells within gradient scaffolds were further supported by both (18)F-fluorine micro-positron emission tomography scanning and in vitro tetracycline labeling. We conclude that the calcium polyphosphate scaffold with gradient pore sizes enhances osteogenic cell differentiation as well as mineralization. The in vivo performance of gradient calcium polyphosphate scaffolds warrants further investigation in animal bone defect models. PMID:26675750

  13. Effect of retinoic acid on murine preosteoblastic MC3T3-E1 cells.

    PubMed

    Nagasawa, Hiroyuki; Takahashi, Shu; Kobayashi, Akira; Tazawa, Hiroshi; Tashima, Yohtalou; Sato, Kozo

    2005-10-01

    Retinoic acid (RA) plays an important role in bone metabolism in vivo through osteoclast activation and bone resorption. Retinoid X-activated receptor beta (RXRbeta) has been implicated in the genetic spinal defect of ossification of the posterior longitudinal ligament (OPLL). In this study, we examined the effects of 9-cis RA and all-trans RA (ATRA) on the proliferation, differentiation, and RXRbeta expression of the murine preosteoblastic cell line MC3T3-E1. Both 9-cis RA and ATRA dose-dependently inhibited the increase in total soluble protein content at concentrations of 10 and 100 nM after 4 and 8 d co-culture with MC3T3-E1 cells. The inhibitory effect of 9-cis RA was slightly stronger than that of ATRA. Histone H4 mRNA expression was dose-dependently suppressed by both RAs on day 1. Alkaline phosphatase activity was increased by both RAs at 10 and 100 nM concentrations on day 4, with 9-cis RA-induced activity slightly stronger than that of ATRA. Osteopontin mRNA expression was increased by both RAs on day 1, but was suppressed on day 4. Bone Gla protein mRNA expression was inhibited by 10 and 100 nM 9-cis RA and by 100 nM ATRA on day 14. RXRbeta mRNA expression was increased by 9-cis RA, an RXRbeta ligand, in a dose-dependent manner. Our results suggested that while both RAs suppressed proliferation and stimulated the maturation of preosteoblastic MC3T3-E1 cells, 9-cis RA was slightly more potent than ATRA. It also appeared that RAs may contribute to the development of heterotopic ossification, including OPLL. PMID:16392701

  14. Calculation of energy levels, {ital E}1 transition amplitudes, and parity violation in francium

    SciTech Connect

    Dzuba, V.A.; Flambaum, V.V.; Sushkov, O.P.

    1995-05-01

    Many-body perturbation theory in the screened Coulomb interaction was used to calculate energy levels, {ital E}1 trransition amplitudes, and the parity-nonconserving (PNC) {ital E}1 amplitude of the 7{ital s}-8{ital s} transition in francium. The method takes into account the core-polarization effect, the second-order correlations, and the three dominating sequences of higher-order correlation diagrams: screening of the electron-electron interaction, particle-hole interaction, and the iterations of the self-energy operator. The result for the PNC amplitude for {sup 223}Fr is {ital E}1(7{ital s}-8{ital s})=(1.59{plus_minus}{similar_to}1%){times}10{sup {minus}10}{ital iea}{sub {ital B}}({minus}{ital Q}{sub {ital W}}/{ital N}), where {ital Q}{sub {ital W}} is the weak charge of the nucleus, {ital N}=136 is the number of neutrons, {ital e}={vert_bar}{ital e}{vert_bar} is the elementary charge, and {ital a}{sub {ital B}} is the Bohr radius. Our prediction for the position of the 8{ital s} energy level of Fr, which has not been measured yet, is 13 110 cm{sup {minus}1} below the limit of the continuous spectrum. The accuracy of the calculations was controlled by comparison with available experimental data and analogous calculations for cesium. It is estimated to be {similar_to}0.1% for the energy levels and {similar_to}1% for the transition amplitudes.

  15. Lysophosphatidic acid induces chemotaxis in MC3T3-E1 osteoblastic cells

    SciTech Connect

    Masiello, Lisa M.; Fotos, Joseph S.; Galileo, Deni S.; Karin, Norm J.

    2006-07-01

    Lysophosphatidic acid (LPA) is a bioactive lipid that has pleiotropic effects on a variety of cell types and enhances the migration of endothelial and cancer cells, but it is not known if this lipid can alter osteoblast motility. We performed transwell migration assays using MC3T3-E1 osteoblastic cells and found LPA to be a potent chemotactic agent. Quantitative time-lapse video analysis of osteoblast migration after wounds were introduced into cell monolayers indicated that LPA stimulated both migration velocity and the average migration distance per cell. LPA also elicited substantial changes in cell shape and actin cytoskeletal structure; lipid-treated cells contained fewer stress fibers and displayed long membrane processes that were enriched in F-actin. Quantitative RT-PCR analysis showed that MC3T3-E1 cells express all four known LPA-specific G protein-coupled receptors (LPA1-LPA4) with a relative mRNA abundance of LPA1 > LPA4 > LPA2 >> LPA3. LPA-induced changes in osteoblast motility and morphology were antagonized by both pertussis toxin and Ki16425, a subtype-specific blocker of LPA1 and LPA3 receptor function. Cell migration in many cell types is linked to changes in intracellular Ca2+. Ki16425 also inhibited LPA-induced Ca2+ signaling in a dose-dependent manner, suggesting a link between LPA-induced Ca2+ transients and osteoblast chemotaxis. Our data show that LPA stimulates MC3T3-E1 osteoblast motility via a mechanism that is linked primarily to the G protein-coupled receptor LPA1.

  16. C-E1 fusion protein synthesized by rubella virus DI RNAs maintained during serial passage

    SciTech Connect

    Tzeng, W.-P.; Frey, Teryl K. . E-mail: tfrey@gsu.edu

    2006-12-20

    Rubella virus (RUB) replicons are derivatives of the RUB infectious cDNA clone that retain the nonstructural open reading frame (NS-ORF) that encodes the replicase proteins but not the structural protein ORF (SP-ORF) that encodes the virion proteins. RUB defective interfering (DI) RNAs contain deletions within the SP-ORF and thus resemble replicons. DI RNAs often retain the 5' end of the capsid protein (C) gene that has been shown to modulate virus-specific RNA synthesis. However, when replicons either with or without the C gene were passaged serially in the presence of wt RUB as a source of the virion proteins, it was found that neither replicon was maintained and DI RNAs were generated. The majority DI RNA species contained in-frame deletions in the SP-ORF leading to a fusion between the 5' end of the C gene and the 3' end of the E1 glycoprotein gene. DI infectious cDNA clones were constructed and transcripts from these DI infectious cDNA clones were maintained during serial passage with wt RUB. The C-E1 fusion protein encoded by the DI RNAs was synthesized and was required for maintenance of the DI RNA during serial passage. This is the first report of a functional novel gene product resulting from deletion during DI RNA generation. Thus far, the role of the C-E1 fusion protein in maintenance of DI RNAs during serial passage remained elusive as it was found that the fusion protein diminished rather than enhanced DI RNA synthesis and was not incorporated into virus particles.

  17. Cooperativity in CYP2E1 metabolism of acetaminophen and styrene mixtures.

    PubMed

    Hartman, Jessica H; Letzig, Lynda G; Roberts, Dean W; James, Laura P; Fifer, E Kim; Miller, Grover P

    2015-10-01

    Risk assessment for exposure to mixtures of drugs and pollutants relies heavily on in vitro characterization of their bioactivation and/or metabolism individually and extrapolation to mixtures assuming no interaction. Herein, we demonstrated that in vitro CYP2E1 metabolic activation of acetaminophen and styrene mixtures could not be explained through the Michaelis-Menten mechanism or any models relying on that premise. As a baseline for mixture studies with styrene, steady-state analysis of acetaminophen oxidation revealed a biphasic kinetic profile that was best described by negative cooperativity (Hill coefficient=0.72). The best-fit mechanism for this relationship involved two binding sites with differing affinities (Ks=830μM and Kss=32mM). Introduction of styrene inhibited that reaction less than predicted by simple competition and thus provided evidence for a cooperative mechanism within the mixture. Likewise, acetaminophen acted through a mixed-type inhibition mechanism to impact styrene epoxidation. In this case, acetaminophen competed with styrene for CYP2E1 (Ki=830μM and Ksi=180μM for catalytic and effector sites, respectively) and resulted in cooperative impacts on binding and catalysis. Based on modeling of in vivo clearance, cooperative interactions between acetaminophen and styrene resulted in profoundly increased styrene activation at low styrene exposure levels and therapeutic acetaminophen levels. Current Michaelis-Menten based toxicological models for mixtures such as styrene and acetaminophen would fail to detect this concentration-dependent relationship. Hence, future studies must assess the role of alternate CYP2E1 mechanisms in bioactivation of compounds to improve the accuracy of interpretations and predictions of toxicity. PMID:26225832

  18. E1 of α-ketoglutarate dehydrogenase defends Mycobacterium tuberculosis against glutamate anaplerosis and nitroxidative stress

    PubMed Central

    Maksymiuk, Christina; Balakrishnan, Anand; Bryk, Ruslana; Rhee, Kyu Y.; Nathan, Carl F.

    2015-01-01

    Enzymes of central carbon metabolism (CCM) in Mycobacterium tuberculosis (Mtb) make an important contribution to the pathogen’s virulence. Evidence is emerging that some of these enzymes are not simply playing the metabolic roles for which they are annotated, but can protect the pathogen via additional functions. Here, we found that deficiency of 2-hydroxy-3-oxoadipate synthase (HOAS), the E1 component of the α-ketoglutarate (α-KG) dehydrogenase complex (KDHC), did not lead to general metabolic perturbation or growth impairment of Mtb, but only to the specific inability to cope with glutamate anaplerosis and nitroxidative stress. In the former role, HOAS acts to prevent accumulation of aldehydes, including growth-inhibitory succinate semialdehyde (SSA). In the latter role, HOAS can participate in an alternative four-component peroxidase system, HOAS/dihydrolipoyl acetyl transferase (DlaT)/alkylhydroperoxide reductase colorless subunit gene (ahpC)-neighboring subunit (AhpD)/AhpC, using α-KG as a previously undescribed source of electrons for reductase action. Thus, instead of a canonical role in CCM, the E1 component of Mtb’s KDHC serves key roles in situational defense that contribute to its requirement for virulence in the host. We also show that pyruvate decarboxylase (AceE), the E1 component of pyruvate dehydrogenase (PDHC), can participate in AceE/DlaT/AhpD/AhpC, using pyruvate as a source of electrons for reductase action. Identification of these systems leads us to suggest that Mtb can recruit components of its CCM for reactive nitrogen defense using central carbon metabolites. PMID:26430237

  19. A comparison of substrate dynamics in human CYP2E1 and CYP2A6

    SciTech Connect

    Harrelson, John P. . E-mail: harrelsonj@pacificu.edu; Henne, Kirk R.; Alonso, Darwin O.V.; Nelson, Sidney D.

    2007-01-26

    Considering the dynamic nature of CYPs, methods that reveal information about substrate and enzyme dynamics are necessary to generate predictive models. To compare substrate dynamics in CYP2E1 and CYP2A6, intramolecular isotope effect experiments were conducted, using deuterium labeled substrates: o-xylene, m-xylene, p-xylene, 2,6-dimethylnaphthalene, and 4,4'-dimethylbiphenyl. Competitive intermolecular experiments were also conducted using d{sub 0}- and d{sub 6}-labeled p-xylene. Both CYP2E1 and CYP2A6 displayed full isotope effect expression for o-xylene oxidation and almost complete suppression for dimethylbiphenyl. Interestingly (k {sub H}/k {sub D}){sub obs} for d{sub 3}-p-xylene oxidation ((k {sub H}/k {sub D}){sub obs} = 6.04 and (k {sub H}/k {sub D}){sub obs} = 5.53 for CYP2E1 and CYP2A6, respectively) was only slightly higher than (k {sub H}/k {sub D}){sub obs} for d{sub 3}-dimethylnaphthalene ((k {sub H}/k {sub D}){sub obs} = 5.50 and (k {sub H}/k {sub D}){sub obs} = 4.96, respectively). One explanation is that in some instances (k {sub H}/k {sub D}){sub obs} values are generated by the presence of two substrates-bound simultaneously to the CYP. Speculatively, if this explanation is valid, then intramolecular isotope effect experiments should be useful in the mechanistic investigation of P450 cooperativity.

  20. Ubiquitination independent of E1 and E2 enzymes by bacterial effectors.

    PubMed

    Qiu, Jiazhang; Sheedlo, Michael J; Yu, Kaiwen; Tan, Yunhao; Nakayasu, Ernesto S; Das, Chittaranjan; Liu, Xiaoyun; Luo, Zhao-Qing

    2016-05-01

    Signalling by ubiquitination regulates virtually every cellular process in eukaryotes. Covalent attachment of ubiquitin to a substrate is catalysed by the E1, E2 and E3 three-enzyme cascade, which links the carboxy terminus of ubiquitin to the ε-amino group of, in most cases, a lysine of the substrate via an isopeptide bond. Given the essential roles of ubiquitination in the regulation of the immune system, it is not surprising that the ubiquitination network is a common target for diverse infectious agents. For example, many bacterial pathogens exploit ubiquitin signalling using virulence factors that function as E3 ligases, deubiquitinases or as enzymes that directly attack ubiquitin. The bacterial pathogen Legionella pneumophila utilizes approximately 300 effectors that modulate diverse host processes to create a permissive niche for its replication in phagocytes. Here we demonstrate that members of the SidE effector family of L. pneumophila ubiquitinate multiple Rab small GTPases associated with the endoplasmic reticulum. Moreover, we show that these proteins are capable of catalysing ubiquitination without the need for the E1 and E2 enzymes. A putative mono-ADP-ribosyltransferase motif critical for the ubiquitination activity is also essential for the role of the SidE family in intracellular bacterial replication in a protozoan host. The E1/E2-independent ubiquitination catalysed by these enzymes is energized by nicotinamide adenine dinucleotide, which activates ubiquitin by the formation of ADP-ribosylated ubiquitin. These results establish that ubiquitination can be catalysed by a single enzyme, the activity of which does not require ATP. PMID:27049943

  1. The Strength-Based Counseling Model

    ERIC Educational Resources Information Center

    Smith, Elsie J.

    2006-01-01

    This article proposes a strength-based model for counseling at-risk youth. The author presents the assumptions, basic concepts, and values of the strength perspective in counseling and offers strength categories as a conceptual model for viewing clients' behavior. Propositions leading toward a theory of strength-based counseling and stages of this

  2. The Strength-Based Counseling Model

    ERIC Educational Resources Information Center

    Smith, Elsie J.

    2006-01-01

    This article proposes a strength-based model for counseling at-risk youth. The author presents the assumptions, basic concepts, and values of the strength perspective in counseling and offers strength categories as a conceptual model for viewing clients' behavior. Propositions leading toward a theory of strength-based counseling and stages of this…

  3. Vinculin expression in MC3T3-E1 cells in response to mechanical stimulus

    PubMed Central

    Cora-Cruz, J.J.; Diffoot-Carlo, N.; Sundaram, P.A.

    2015-01-01

    Loading frequency is known to influence the expression of the focal adhesions of the adherent cells. A small cyclical tensile force was transmitted to mouse pre-osteoblast MC3T3-E1 cells through PDMS substrates of varying stiffness. Changes in cell behavior with respect to proliferation and characteristics of focal adhesions were quantified through immunofluorescence labeling of vinculin. Amount of inactive vinculin was higher on substrates subjected to cyclic stimulation when compared with the results of the static substrates, whereas the number and area of focal adhesion points underwent a reduction. Inactive vinculin appears as a cloud in the cytoplasm in the vicinity of the nucleus. PMID:26858974

  4. Inhibition of cytochrome P4502E1 by chlormethiazole attenuated acute ethanol-induced fatty liver.

    PubMed

    Chen, Yang-Yang; Zhang, Cui-Li; Zhao, Xiu-Lan; Xie, Ke-Qin; Zeng, Tao

    2014-08-24

    Cytochrome P4502E1 (CYP2E1) has been demonstrated to play crucial roles in chronic ethanol-induced fatty liver, while its role in acute ethanol-induced fatty liver remains unclear. The current study was designed to evaluate the effects of chlormethiazole (CMZ), a specific inhibitor of CYP2E1, on acute ethanol-induced fatty liver, and to explore the mechanisms. Mice were pretreated with single dose of CMZ (50mg/kg body weight) by intraperitoneal injection or equal volume of saline, and then exposed to three doses of ethanol (5g/kg body weight, 25%, w/v) by gavage with 12h intervals. The mice were sacrificed at 4h after the last ethanol dosing. It was found that CMZ significantly attenuated acute ethanol-induced increase of the hepatic and serum triglyceride levels, and reduced fat droplets accumulation in mice liver. Acute ethanol-induced increase of the hepatic malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) levels (two biomarkers for oxidative stress) and decrease of glutathione (GSH) level was significantly suppressed by CMZ. CMZ also suppressed ethanol-induced decline of serum adiponectin level, but did not significantly affect the serum tumor necrosis factor-α (TNF-α) and ethanol levels. Furthermore, a significant decline of p62 protein level was observed in CMZ/ethanol group mice liver compared with that of the ethanol group mice. However, acute ethanol-induced increase of peroxisome proliferator-activated receptor α (PPAR-α) protein level was suppressed by CMZ, while the protein levels of sterol regulatory element-binding protein-1c (SREBP-1) and diacylglycerol acyltransferase 2 (DGAT2) were not significantly affected by ethanol or CMZ. Collectively, the results of the current study demonstrated that CMZ could effectively attenuate acute ethanol-induced fatty liver possibly by suppressing oxidative stress and adiponectin decline, and activating autophagy, which suggest that CYP2E1 might also play important roles in acute ethanol-induced fatty liver. PMID:25162931

  5. Calculation of radiative corrections to E1 matrix elements in the neutral alkali metals

    SciTech Connect

    Sapirstein, J.; Cheng, K.T.

    2005-02-01

    Radiative corrections to E1 matrix elements for ns-np transitions in the alkali-metal atoms lithium through francium are evaluated. They are found to be small for the lighter alkali metals but significantly larger for the heavier alkali metals, and in the case of cesium much larger than the experimental accuracy. The relation of the matrix element calculation to a recent decay rate calculation for hydrogenic ions is discussed, and application of the method to parity nonconservation in cesium is described.

  6. GALILÉE-1: a validation and processing system for ENDF-6 and GND evaluations

    NASA Astrophysics Data System (ADS)

    Coste-Delclaux, Mireille; Jouanne, Cédric; Moreau, Frédéric; Mounier, Claude

    2016-03-01

    GALILÉE-1 is the new validation and processing system for evaluated data, developed at CEA. This system can handle evaluations stored either in the ENDF-6 format or in the new General Nuclear Data (GND) format. It consists of various components respectively dedicated to read/write the evaluated data whatever the format is, to diagnose inconsistencies in the evaluated data and to provide continuous-energy and multigroup data as well as probability tables for transport and depletion codes. All these components are written in C++ language and share the same objects. This paper describes the state of progress of the various parts of the system and gives some illustrations.

  7. Observational Results of the ChangE-1 Solar X-Ray Monitor

    NASA Astrophysics Data System (ADS)

    Cui, X. Z.; Wang, H. Y.; Peng, W. X.; Zhang, C. M.; Liang, X. H.; Wang, J. Z.; Gao, M.; Yang, J. W.; Cao, X. L.; Zhang, J. Y.; Wu, M. Y.; Chang, J.; Sun, H. X.; OuYang, Z. Y.; Zhou, Y. L.; Li, C. L.

    2014-05-01

    We present the primary observations of the Solar X-ray Monitor (SXM) payload onboard the ChangE-1 lunar exploration satellite, which was launched on 24 October 2007. The SXM payload uses a solid-state silicon P-I-N photo-diode (Si-PIN) whose dynamic energy ranges from 1 keV to 10 keV. The long-term integrated spectra at different solar-activity levels as observed by the SXM are presented. By fitting these spectra with an optically thin plasma model, the two-minute temperature variation of the solar coronal plasma during a solar flare is also presented.

  8. Bortezomib alleviates drug-induced liver injury by regulating CYP2E1 gene transcription

    PubMed Central

    PARK, WOO-JAE; KIM, SO-YEON; KIM, YE-RYUNG; PARK, JOO-WON

    2016-01-01

    Acute liver failure, i.e., the fatal deterioration of liver function, is the most common indication that emergency liver transplantation is necessary. Moreover, in the USA, drug-induced liver injury (DILI), including acetaminophen (APAP)-induced hepatotoxicity, is the main cause of acute liver failure. Matching a donor for liver transplantation is extremely difficult, and thus the development of a novel therapy for DILI is urgently needed. Following recent approval by the FDA of the proteasomal inhibitor bortezomib, its therapeutic effects on various human diseases, including solid and hematologic malignancies, have been validated. However, the specific action of proteasomal inhibition in cases of DILI had not been elucidated prior to this study. To examine the effects of proteasomal inhibition in DILI experimentally, male C56Bl/6 mice were injected with 1 mg bortezomib/kg before APAP treatment. Bortezomib not only alleviated APAP-induced hepatotoxicity in a time- and dose-dependent manner, it also alleviated CCl4- and thioacetamide-induced hepatotoxicity. We also noted that bortezomib significantly reduced cytochrome P450 2E1 (CYP2E1) expression and activity in the liver, which was accompanied by the induction of endoplasmic reticulum (ER) stress. In addition, bortezomib decreased hepatocyte nuclear factor-1α-induced promoter activation of CYP2E1 in Hep3B cells. By contrast, another proteasome inhibitor, MG132, did not cause ER stress and did not markedly affect CYP2E1 enzyme activity. Liver injury induced by APAP was aggravated by MG132, possibly via elevation of connexin 32 expression. This study suggests that proteasome inhibition has different effects in cases of DILI depending on the specific inhibitor being used. Furthermore, results from the mouse model indicated that bortezomib, but not MG132, was effective in alleviating DILI. ER stress induced by proteasome inhibition has previously been shown to exert various effects on DILI patients, and thus each available proteasomal inhibitor should be evaluated individually in order to determine its potential for clinical application. PMID:26797017

  9. Vinculin expression in MC3T3-E1 cells in response to mechanical stimulus.

    PubMed

    Cora-Cruz, J J; Diffoot-Carlo, N; Sundaram, P A

    2016-03-01

    Loading frequency is known to influence the expression of the focal adhesions of the adherent cells. A small cyclical tensile force was transmitted to mouse pre-osteoblast MC3T3-E1 cells through PDMS substrates of varying stiffness. Changes in cell behavior with respect to proliferation and characteristics of focal adhesions were quantified through immunofluorescence labeling of vinculin. Amount of inactive vinculin was higher on substrates subjected to cyclic stimulation when compared with the results of the static substrates, whereas the number and area of focal adhesion points underwent a reduction. Inactive vinculin appears as a cloud in the cytoplasm in the vicinity of the nucleus. PMID:26858974

  10. The relation between the photonuclear E1 sum rule and the effective orbital g-factor

    NASA Astrophysics Data System (ADS)

    Bentz, Wolfgang; Arima, Akito

    2004-05-01

    The connection between the enhancement factor (1+ κ) of the photonuclear E1 sum rule and the orbital angular momentum g-factor ( gℓ) of a bound nucleon is investigated in the framework of the Landau-Migdal theory for isospin asymmetric nuclear matter. Special emphasis is put on the role of gauge invariance to establish the κ- gℓ relation. By identifying the physical processes which are taken into account in κ and gℓ, the validity and limitations of this relation will be discussed. The connections to the collective excitations and to nuclear Compton scattering are also shown.

  11. E1 reaction-induced synthesis of hydrophilic oxide nanoparticles in a non-hydrophilic solvent

    NASA Astrophysics Data System (ADS)

    Hu, Ming-Jun; Gao, Jiefeng; Yang, Shiliu; Dong, Yucheng; Ping Wong, Julia Shuk; Xu, Jiaju; Shan, Guangcun; Li, Robert K. Y.

    2012-09-01

    In this paper, tert-amyl alcohol was employed to directly react with metal chlorides for the preparation of oxide nanoparticles. Some typical metal oxide or hydroxides with different morphologies, such as TiO2 nanoparticles, TiO2 nanorods, FeOOH nanowires, Fe2O3 nanoparticles, and SnO2 nanoparticles, can be easily fabricated through such simple chemical reactions. E1 reaction was found to play the leading role in the synthesis of metal oxides attributed to better stability of tertiary carbocations in tert-amyl alcohol and the strong interaction of metal chlorides with hydroxyl groups that results in the easy dissociation of carbon-oxygen bonds in tert-amyl alcohol. SN1 reaction can also occur in certain reactions due to nucleophilic substitution of chloride ions for hydroxyl groups. As-prepared metal oxides show good compatibility with an aqueous system while they were synthesized in a non-hydrophilic solvent probably attributed to the specific E1 reaction mechanism involving the generation of water, and can be directly incorporated into an aqueous soluble polymer, such as PVA, to exhibit many promising applications.In this paper, tert-amyl alcohol was employed to directly react with metal chlorides for the preparation of oxide nanoparticles. Some typical metal oxide or hydroxides with different morphologies, such as TiO2 nanoparticles, TiO2 nanorods, FeOOH nanowires, Fe2O3 nanoparticles, and SnO2 nanoparticles, can be easily fabricated through such simple chemical reactions. E1 reaction was found to play the leading role in the synthesis of metal oxides attributed to better stability of tertiary carbocations in tert-amyl alcohol and the strong interaction of metal chlorides with hydroxyl groups that results in the easy dissociation of carbon-oxygen bonds in tert-amyl alcohol. SN1 reaction can also occur in certain reactions due to nucleophilic substitution of chloride ions for hydroxyl groups. As-prepared metal oxides show good compatibility with an aqueous system while they were synthesized in a non-hydrophilic solvent probably attributed to the specific E1 reaction mechanism involving the generation of water, and can be directly incorporated into an aqueous soluble polymer, such as PVA, to exhibit many promising applications. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr32255b

  12. M1 {gamma} Strength for Zirconium Nuclei in the Photoneutron Channel

    SciTech Connect

    Utsunomiya, H.; Kondo, T.; Kaihori, T.; Makinaga, A.; Akimune, H.; Yamagata, T.; Goriely, S.; Goko, S.; Toyokawa, H.; Matsumoto, T.; Harano, H.; Hohara, S.; Lui, Y.-W.; Hilaire, S.; Peru, S.; Koning, A. J.

    2008-04-25

    Photoneutron cross sections were measured for {sup 91}Zr, {sup 92}Zr, and {sup 94}Zr near the neutron separation energy with quasimonochromatic {gamma} rays. The data exhibit some extra components around the neutron threshold. A coherent analysis of the photoneutron data for {sup 92}Zr together with the neutron capture on {sup 91}Zr based on the microscopic Hartree-Fock-Bogoliubov plus quasiparticle random-phase approximation model for the E1 strength has revealed the presence of an M1 resonance at 9 MeV. The microscopic approach systematically shows the same M1 strength in the photoneutron cross section for {sup 91}Zr and {sup 94}Zr. The total M1 strength is about 75% larger than the strength predicted by the systematics, being qualitatively consistent with the giant M1 resonance observed in the inelastic proton scattering.

  13. Role for intestinal CYP2E1 in alcohol-induced circadian gene-mediated intestinal hyperpermeability

    PubMed Central

    Voigt, Robin M.; Shaikh, Maliha; Tang, Yueming; Cederbaum, Arthur I.; Turek, Fred W.; Keshavarzian, Ali

    2013-01-01

    We have shown that alcohol increases Caco-2 intestinal epithelial cell monolayer permeability in vitro by inducing the expression of redox-sensitive circadian clock proteins CLOCK and PER2 and that these proteins are necessary for alcohol-induced hyperpermeability. We hypothesized that alcohol metabolism by intestinal Cytochrome P450 isoform 2E1 (CYP2E1) could alter circadian gene expression (Clock and Per2), resulting in alcohol-induced hyperpermeability. In vitro Caco-2 intestinal epithelial cells were exposed to alcohol, and CYP2E1 protein, activity, and mRNA were measured. CYP2E1 expression was knocked down via siRNA and alcohol-induced hyperpermeability, and CLOCK and PER2 protein expression were measured. Caco-2 cells were also treated with alcohol or H2O2 with or without N-acetylcysteine (NAC) anti-oxidant, and CLOCK and PER2 proteins were measured at 4 or 2 h. In vivo Cyp2e1 protein and mRNA were also measured in colon tissue from alcohol-fed mice. Alcohol increased CYP2E1 protein by 93% and enzyme activity by 69% in intestinal cells in vitro. Alcohol feeding also increased mouse colonic Cyp2e1 protein by 73%. mRNA levels of Cyp2e1 were not changed by alcohol in vitro or in mouse intestine. siRNA knockdown of CYP2E1 in Caco-2 cells prevented alcohol-induced hyperpermeability and induction of CLOCK and PER2 proteins. Alcohol-induced and H2O2-induced increases in intestinal cell CLOCK and PER2 were significantly inhibited by treatment with NAC. We concluded that our data support a novel role for intestinal CYP2E1 in alcohol-induced intestinal hyperpermeability via a mechanism involving CYP2E1-dependent induction of oxidative stress and upregulation of circadian clock proteins CLOCK and PER2. PMID:23660503

  14. The cognition-enhancing activity of E1R, a novel positive allosteric modulator of sigma-1 receptors

    PubMed Central

    Zvejniece, L; Vavers, E; Svalbe, B; Vilskersts, R; Domracheva, I; Vorona, M; Veinberg, G; Misane, I; Stonans, I; Kalvinsh, I; Dambrova, M

    2014-01-01

    Background and Purpose Here, we describe the in vitro and in vivo effects of (4R,5S)-2-(5-methyl-2-oxo-4-phenyl-pyrrolidin-1-yl)-acetamide (E1R), a novel positive allosteric modulator of sigma-1 receptors. Experimental Approach E1R was tested for sigma receptor binding activity in a [3H](+)-pentazocine assay, in bradykinin (BK)-induced intracellular Ca2+ concentration ([Ca2+]i) assays and in an electrically stimulated rat vas deferens model. E1R's effects on cognitive function were tested using passive avoidance (PA) and Y-maze tests in mice. A selective sigma-1 receptor antagonist (NE-100), was used to study the involvement of the sigma-1 receptor in the effects of E1R. The open-field test was used to detect the effects of E1R on locomotion. Key Results Pretreatment with E1R enhanced the selective sigma-1 receptor agonist PRE-084's stimulating effect during a model study employing electrically stimulated rat vasa deferentia and an assay measuring the BK-induced [Ca2+]i increase. Pretreatment with E1R facilitated PA retention in a dose-related manner. Furthermore, E1R alleviated the scopolamine-induced cognitive impairment during the PA and Y-maze tests in mice. The in vivo and in vitro effects of E1R were blocked by treatment with the selective sigma-1 receptor antagonist NE-100. E1R did not affect locomotor activity. Conclusion and Implications E1R is a novel 4,5-disubstituted derivative of piracetam that enhances cognition and demonstrates efficacy against scopolamine-induced cholinergic dysfunction in mice. These effects are attributed to its positive modulatory action on the sigma-1 receptor and this activity may be relevant when developing new drugs for treating cognitive symptoms related to neurodegenerative diseases. PMID:24490863

  15. Bax inhibitor 1 regulates ER-stress-induced ROS accumulation through the regulation of cytochrome P450 2E1.

    PubMed

    Kim, Hyung-Ryong; Lee, Geum-Hwa; Cho, Eun Yi; Chae, Soo-Wan; Ahn, Taeho; Chae, Han-Jung

    2009-04-15

    This study investigated the molecular mechanism by which Bax inhibitor 1 (BI1) abrogates the accumulation of reactive oxygen species (ROS) in the endoplasmic reticulum (ER). Electron uncoupling between NADPH-dependent cytochrome P450 reductase (NPR) and cytochrome P450 2E1 (P450 2E1) is a major source of ROS on the ER membrane. ER stress produced ROS accumulation and lipid peroxidation of the ER membrane, but BI1 reduced this accumulation. Under ER stress, expression of P450 2E1 in control cells was upregulated more than in BI1-overexpressing cells. In control cells, inhibiting P450 2E1 through chemical or siRNA approaches suppressed ROS accumulation, ER membrane lipid peroxidation and the resultant cell death after ER stress. However, it had little effect in BI1-overexpressing cells. In addition, BI1 knock down also increased ROS accumulation and expression of P450 2E1. In a reconstituted phospholipid membrane containing purified BI1, NPR and P450 2E1, BI1 dose-dependently decreased the production of ROS. BI1 bound to NPR with higher affinity than P450 2E1. Furthermore, BI1 overexpression reduced the interaction of NPR and P450 2E1, and decreased the catalytic activity of P450 2E1, suggesting that the flow of electrons from NPR to P450 2E1 can be modulated by BI1. In summary, BI1 reduces the accumulation of ROS and the resultant cell death through regulating P450 2E1. PMID:19339548

  16. Human Placental Lactogen Induces CYP2E1 Expression via PI 3-Kinase Pathway in Female Human Hepatocytes

    PubMed Central

    Lee, Jin Kyung; Chung, Hye Jin; Fischer, Liam; Fischer, James; Gonzalez, Frank J.

    2014-01-01

    The state of pregnancy is known to alter hepatic drug metabolism. Hormones that rise during pregnancy are potentially responsible for the changes. Here we report the effects of prolactin (PRL), placental lactogen (PL), and growth hormone variant (GH-v) on expression of major hepatic cytochromes P450 expression and a potential molecular mechanism underlying CYP2E1 induction by PL. In female human hepatocytes, PRL and GH-v showed either no effect or small and variable effects on mRNA expression of CYP1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1, 3A4, and 3A5. On the other hand, PL increased expression level of CYP2E1 mRNA with corresponding increases in CYP2E1 protein and activity levels. Results from hepatocytes and HepaRG cells indicate that PL does not affect the expression or activity of HNF1α, the known transcriptional activator of basal CYP2E1 expression. Furthermore, transient transfection studies and Western blot results showed that STAT signaling, the previously known mediator of PL actions in certain tissues, does not play a role in CYP2E1 induction by PL. A chemical inhibitor of PI3-kinase signaling significantly repressed the CYP2E1 induction by PL in human hepatocytes, suggesting involvement of PI3-kinase pathway in CYP2E1 regulation by PL. CYP2E1-humanized mice did not exhibit enhanced CYP2E1 expression during pregnancy, potentially because of interspecies differences in PL physiology. Taken together, these results indicate that PL induces CYP2E1 expression via PI3-kinase pathway in human hepatocytes. PMID:24408518

  17. Mutation and evolutionary analyses identify NR2E1-candidate-regulatory mutations in humans with severe cortical malformations

    PubMed Central

    Kumar, R A; Leach, S; Bonaguro, R; Chen, J; Yokom, D W; Abrahams, B S; Seaver, L; Schwartz, C E; Dobyns, W; Brooks-Wilson, A; Simpson, E M

    2007-01-01

    Nuclear receptor 2E1 (NR2E1) is expressed in human fetal and adult brains; however, its role in human brain–behavior development is unknown. Previously, we have corrected the cortical hypoplasia and behavioral abnormalities in Nr2e1−/− mice using a genomic clone spanning human NR2E1, which bolsters the hypothesis that NR2E1 may similarly play a role in human cortical and behavioral development. To test the hypothesis that humans with abnormal brain–behavior development may have null or hypomorphic NR2E1 mutations, we undertook the first candidate mutation screen of NR2E1 by sequencing its entire coding region, untranslated, splice site, proximal promoter and evolutionarily conserved non-coding regions in 56 unrelated patients with cortical disorders, namely microcephaly. We then genotyped the candidate mutations in 325 unrelated control subjects and 15 relatives. We did not detect any coding region changes in NR2E1; however, we identified seven novel candidate regulatory mutations that were absent from control subjects. We used in silico tools to predict the effects of these candidate mutations on neural transcription factor binding sites (TFBS). Four candidate mutations were predicted to alter TFBS. To facilitate the present and future studies of NR2E1, we also elucidated its molecular evolution, genetic diversity, haplotype structure and linkage disequilibrium by sequencing an additional 94 unaffected humans representing Africa, the Americas, Asia, Europe, the Middle East and Oceania, as well as great apes and monkeys. We detected strong purifying selection, low genetic diversity, 21 novel polymorphisms and five common haplotypes at NR2E1. We conclude that protein-coding changes in NR2E1 do not contribute to cortical and behavioral abnormalities in the patients examined here, but that regulatory mutations may play a role. PMID:17054721

  18. Crystal structure of hyperthermophilic esterase EstE1 and the relationship between its dimerization and thermostability properties

    PubMed Central

    Byun, Jung-Sue; Rhee, Jin-Kyu; Kim, Nam Doo; Yoon, JeongHyeok; Kim, Dong-Uk; Koh, Eunhee; Oh, Jong-Won; Cho, Hyun-Soo

    2007-01-01

    Background EstE1 is a hyperthermophilic esterase belonging to the hormone-sensitive lipase family and was originally isolated by functional screening of a metagenomic library constructed from a thermal environmental sample. Dimers and oligomers may have been evolutionally selected in thermophiles because intersubunit interactions can confer thermostability on the proteins. The molecular mechanisms of thermostabilization of this extremely thermostable esterase are not well understood due to the lack of structural information. Results Here we report for the first time the 2.1-Å resolution crystal structure of EstE1. The three-dimensional structure of EstE1 exhibits a classic α/β hydrolase fold with a central parallel-stranded beta sheet surrounded by alpha helices on both sides. The residues Ser154, Asp251, and His281 form the catalytic triad motif commonly found in other α/β hydrolases. EstE1 exists as a dimer that is formed by hydrophobic interactions and salt bridges. Circular dichroism spectroscopy and heat inactivation kinetic analysis of EstE1 mutants, which were generated by structure-based site-directed mutagenesis of amino acid residues participating in EstE1 dimerization, revealed that hydrophobic interactions through Val274 and Phe276 on the β8 strand of each monomer play a major role in the dimerization of EstE1. In contrast, the intermolecular salt bridges contribute less significantly to the dimerization and thermostability of EstE1. Conclusion Our results suggest that intermolecular hydrophobic interactions are essential for the hyperthermostability of EstE1. The molecular mechanism that allows EstE1 to endure high temperature will provide guideline for rational design of a thermostable esterase/lipase using the lipolytic enzymes showing structural similarity to EstE1. PMID:17625021

  19. Mesenchymal Stem Cell Durotaxis Depends on Substrate Stiffness Gradient Strength

    PubMed Central

    Vincent, Ludovic G.; Choi, Yu Suk; Alonso-Latorre, Baldomero; del Álamo, Juan C.; Engler, Adam J.

    2013-01-01

    Mesenchymal stem cells (MSCs) respond to niche elasticity, which varies between and within tissues. Stiffness gradients result from pathological conditions but also occur through normal variation, e.g. muscle. MSCs undergo directed migration even in response to shallow stiffness gradients before differentiating. More refined gradients of both stiffness range and strength are needed to better understand mechanical regulation of migration in normal and disease pathologies. We describe polyacrylamide stiffness gradient fabrication using three distinct systems that generate stiffness gradients of physiological (1 Pa/µm), pathological (10 Pa/µm), and step (≥ 100Pa/um) strength spanning physiologically relevant stiffness for most soft tissue, i.e. 1–12 kPa. MSCs migrated to the stiffest region for each gradient. Time-lapse microscopy revealed that migration velocity scaled directly with gradient strength. Directed migration was reduced in the presence of the contractile agonist lysophosphatidic acid (LPA) and cytoskeletal-perturbing drugs nocodazole and cytochalasin; LPA- and nocodazole-treated cells remained spread and protrusive, while cytochalasin-treated cells did not. Untreated and nocodazole-treated cells spread in a similar manner, but nocodazole-treated cells had greatly diminished traction forces. These data suggest that actin is required for migration whereas microtubules are required for directed migration. The data also imply that in vivo, MSCs may have a more significant contribution to repairs in stiffer regions where they may preferentially accumulate. PMID:23390141

  20. Phytocomponent p-hydroxycinnamic acid stimulates mineralization in osteoblastic MC3T3-E1 cells.

    PubMed

    Yamaguchi, Masayoshi; Lai, Ying Ling; Uchiyama, Satoshi; Nakagawa, Taeko

    2008-09-01

    Phytocomponent p-hydroxycinnamic acid (HCA) has been shown to have stimulatory effects on bone calcification and inhibitory effects on bone resorption in rat femoral tissues in vitro. Whether HCA has a stimulatory effect on mineralization in osteoblastic cells is unknown. This study was undertaken to determine the effect of HCA on mineralization in osteoblastic MC3T3-E1 cells in vitro. Cells were cultured for 72 h in a minimum essential medium (alpha-MEM) containing 10% fetal bovine serum (FBS), and the cells with subconfluency were changed to a medium containing either vehicle or HCA (10(-7)-10(-5) M) without FBS. Culture with HCA (10(-7)-10(-5) M) did not have a significant effect on cell proliferation and cell death. Deoxyribonucleic acid (DNA) content in osteoblastic cells was significantly increased after culture with HCA (10(-6) or 10(-5) M) for 48 or 72 h. Alkaline phosphatase activity in osteoblastic cells was significantly increased after culture with HCA (10(-7)-10(-5) M) for 24, 48, or 72 h. The results with Alizarin red staining for calcium showed that mineralization was significantly stimulated after culture with HCA (10(-8)-10(-5) M) for 7, 14, or 21 days. This study demonstrates that HCA has stimulatory effects on mineralization in osteoblastic MC3T3-E1 cells. PMID:18698486

  1. CD36 is a co-receptor for hepatitis C virus E1 protein attachment.

    PubMed

    Cheng, Jun-Jun; Li, Jian-Rui; Huang, Meng-Hao; Ma, Lin-Lin; Wu, Zhou-Yi; Jiang, Chen-Chen; Li, Wen-Jing; Li, Yu-Huan; Han, Yan-Xing; Li, Hu; Chen, Jin-Hua; Wang, Yan-Xiang; Song, Dan-Qing; Peng, Zong-Gen; Jiang, Jian-Dong

    2016-01-01

    The cluster of differentiation 36 (CD36) is a membrane protein related to lipid metabolism. We show that HCV infection in vitro increased CD36 expression in either surface or soluble form. HCV attachment was facilitated through a direct interaction between CD36 and HCV E1 protein, causing enhanced entry and replication. The HCV co-receptor effect of CD36 was independent of that of SR-BI. CD36 monoclonal antibodies neutralized the effect of CD36 and reduced HCV replication. CD36 inhibitor sulfo-N-succinimidyl oleate (SSO), which directly bound CD36 but not SR-BI, significantly interrupted HCV entry, and therefore inhibited HCV replication. SSO's antiviral effect was seen only in HCV but not in other viruses. SSO in combination with known anti-HCV drugs showed additional inhibition against HCV. SSO was considerably safe in mice. Conclusively, CD36 interacts with HCV E1 and might be a co-receptor specific for HCV entry; thus, CD36 could be a potential drug target against HCV. PMID:26898231

  2. Maintenance and activation of Cyp2e-1 gene expression in mouse hepatocytes in primary culture.

    PubMed

    Sakurai, J; Funae, Y; Nemoto, N

    1996-08-21

    The expression of Cyp2e-1 mRNA and protein was investigated in the C57BL/6NCrj mouse hepatocytes in primary culture, as well as liver and kidney. The mRNA and protein expression in the liver was in the same range in both sexes and was not affected by orchiectomy or ovariectomy. The mRNA expression was enhanced in the kidney of ovariectomized mice, in which the protein contents were not influenced. Orchiectomy decreased the expression of both mRNA and protein. When the hepatocytes were transferred to primary culture, the amounts of the mRNA were not changed within 24 h and about half remained by day 3. However, the expression was low thereafter. The expression of the protein gradually decreased after the start of culture. Dexamethasone showed a potential as an inducer at more than 10(-8) M. Sex hormones increased the expression of this P-450 species a little in culture, but growth hormone did not. These observations indicated that glucocorticoid hormone plays a role in modifying expression of Cyp2e-1 and that the mouse hepatocyte culture is useful for examining its regulation mechanism. PMID:8781547

  3. [Genetic polymorphism of cytochrome P450 2E1 and the risk of nasopharyngeal carcinoma].

    PubMed

    Ben Chaaben, Arij; Abaza, Hajer; Douik, Hayet; Chaouch, Leila; Ayari, Fayza; Ouni, Nesrine; Mamoghli, Tasnim; Ben Guezella, Dorra; Mejri, Rachida; Harzallah, Latifa; Guemira, Fethi

    2015-12-01

    Cytochrome P450 2E1 (CYP2E1) is a detoxifying enzyme that belongs to the phase I metabolism of xenobiotics. This enzyme is encoded by a highly polymorphic gene whose common polymorphism corresponds to the substitution of cytosine (C) and thymine (T) at position -1019 (rs2031920). This polymorphism has been identified in several cancers including nasopharyngeal cancer (NPC). The study involved 124 patients with nasopharyngeal carcinoma, compared with 166 healthy controls. The presence or absence of the polymorphism is determined by PCR-RFLP. The frequency comparison between the two groups is determined by the χ(2) test. The analysis of our results showed a significant difference between the two groups regarding the mutant genotype (C2/C2) (5% vs. 0.5%, P=0.04) and has a risk factor for NPC in Tunisia (OR=8.39; CI 95% [0.99-388.1]). Also, the C2 allele was significantly associated with the group of patients than the control group (6% vs. 2%, P=0.016) and increased three times the risk of NPC in Tunisia (OR=2.99, CI 95% [1.12-8.79]). Our results confirm the results reported in other populations and emphasize the importance of the involvement of this gene in the development of detoxification of the NPC, which seems more and more strongly associated with environmental factors. PMID:26582733

  4. First Selenodetic Result from Chang'E-1 Lunar Orbiter Mission

    NASA Astrophysics Data System (ADS)

    Ping, Jinsong; Huang, Qian; Yan, Jianguo; Shi, Xian; Wang, Guangli; Sha, Kai; Tang, Geshi; Chen, Ming; Cao, Jianfeng

    The first Chinese lunar orbiter, Chang'E-1, was sent to the circle polar orbit of the moon on Nov. 5. During the first month mission checkout period and the following normal mission period, it has been tracked using Range and RR and VLBI method. The tracking data may contribute to both of the OD and the POD of the mission. At the same time, the laser altimeter has been operated to measure the topography of the moon with very high accuracy and spatial resolution. Using the domestic tracking data, POD of Chang'E-1 mission was carried out. It is expected to merge the tracking data with other missions so as to improve the lunar global gravity field. By combining the POD data and attitude information as well as other necessary telemetry messages, the topography of the moon was renewed. Based on the renewed lunar DEM information and the lunar gravity field, the lunar crastial sphere and the Moho surface have been estimated using a downward continuation analyzing method. It may contribute to the selenodetic researches by combining with the historic data.

  5. Prostaglandin E1 increases survival with extended anhepatic phase during liver transplantation.

    PubMed Central

    Xu, H S; Rosenlof, L K; Pruett, T L; Jones, R S

    1994-01-01

    OBJECTIVE: The authors investigated the intraoperative treatment effects of Prostaglandin E1 (PGE1) for extension of the anhepatic phase and improvement of survival in a rat liver transplant model. BACKGROUND: Cross-clamping the inferior vena cava and the portal vein during liver transplantation causes severe pathophysiologic changes during surgery. The time of the anhepatic phase is strictly limited and results in a very tenuous period during the liver transplant operation. METHODS: Prostaglandin E1 was infused at 0.5 microgram/kg/min into five subgroups of rats with 20, 30, 40, 60, and 80 minutes of anhepatic phase during transplantation. Bile secretion, serum aspartate transaminase (AST), lactic dehydrogenase (LDH), and blood gas analysis were studied in the 30-minute subgroup. The results were compared with the sham-operated and control groups. RESULTS: Intraoperative treatment with PGE1 extended the maximal anhepatic phase from 30 minutes in the sham-operated group up to 80 minutes, and increased survival. Significant changes in the PGE1 treated rats in the 30-minute subgroup included an increase of bile flow and bile salt output and decrease of AST and LDH activities after surgery. Blood gas analysis showed a decrease in acidosis and hypercarbia at the end of the anhepatic phase. CONCLUSIONS: The PGE1 treatment increased survival with extended anhepatic phase during rat liver transplantation. The beneficial effects can be attributed to its biologic activities. PMID:8024359

  6. Highly charged ions with E1, M1, and E2 transitions within laser range

    NASA Astrophysics Data System (ADS)

    Berengut, J. C.; Dzuba, V. A.; Flambaum, V. V.; Ong, A.

    2012-08-01

    Level crossings in the ground state of ions occur when the nuclear charge Z and ion charge Zion are varied along an isoelectronic sequence until the two outermost shells are nearly degenerate. We examine all available level crossings in the periodic table for both near-neutral ions and highly charged ions (HCIs). Normal E1 transitions in HCIs are in x-ray range; however, level crossings allow for optical electromagnetic transitions that could form the reference transition for high-accuracy atomic clocks. Optical E1 (due to configuration mixing), M1, and E2 transitions are available in HCIs near level crossings. We present scaling laws for energies and amplitudes that allow us to make simple estimates of systematic effects of relevance to atomic clocks. HCI clocks could have some advantages over existing optical clocks because certain systematic effects are reduced; for example, they can have much smaller thermal shifts. Other effects such as fine-structure and hyperfine splitting are much larger in HCIs, which can allow for richer spectra. HCIs are excellent candidates for probing variations in the fine-structure constant α in atomic systems as there are transitions with the highest sensitivity to α variation.

  7. Construction of an adenovirus type 7a E1A- vector.

    PubMed Central

    Abrahamsen, K; Kong, H L; Mastrangeli, A; Brough, D; Lizonova, A; Crystal, R G; Falck-Pedersen, E

    1997-01-01

    A strategy for constructing replication-defective adenovirus vectors from non-subgroup C viruses has been successfully demonstrated with adenovirus type 7 strain a (Ad7a) as the prototype. An E1A-deleted Ad7a reporter virus expressing the chloramphenicol acetyltransferase (CAT) gene from the cytomegalovirus promoter enhancer was constructed with DNA fragments isolated from Ad7a, an Ad7a recombination reporter plasmid, and the 293 cell line. The Ad7a-CAT virus particle transduces A549 cells as efficiently as Ad5-based vectors. Intravenous infections in a murine model indicate that the Ad7a-CAT virus infects a variety of tissues, with maximal levels of CAT gene expression found in the liver. The duration of Ad7a-CAT transgene expression in the liver was maximally maintained 2 weeks postinfection, with a decline to baseline activity by the week 4 postinfection. Ad7a-CAT represents the first example of a non-subgroup C E1A- adenovirus gene transfer vector. PMID:9343264

  8. CD36 is a co-receptor for hepatitis C virus E1 protein attachment

    PubMed Central

    Cheng, Jun-Jun; Li, Jian-Rui; Huang, Meng-Hao; Ma, Lin-Lin; Wu, Zhou-Yi; Jiang, Chen-Chen; Li, Wen-Jing; Li, Yu-Huan; Han, Yan-Xing; Li, Hu; Chen, Jin-Hua; Wang, Yan-Xiang; Song, Dan-Qing; Peng, Zong-Gen; Jiang, Jian-Dong

    2016-01-01

    The cluster of differentiation 36 (CD36) is a membrane protein related to lipid metabolism. We show that HCV infection in vitro increased CD36 expression in either surface or soluble form. HCV attachment was facilitated through a direct interaction between CD36 and HCV E1 protein, causing enhanced entry and replication. The HCV co-receptor effect of CD36 was independent of that of SR-BI. CD36 monoclonal antibodies neutralized the effect of CD36 and reduced HCV replication. CD36 inhibitor sulfo-N-succinimidyl oleate (SSO), which directly bound CD36 but not SR-BI, significantly interrupted HCV entry, and therefore inhibited HCV replication. SSO’s antiviral effect was seen only in HCV but not in other viruses. SSO in combination with known anti-HCV drugs showed additional inhibition against HCV. SSO was considerably safe in mice. Conclusively, CD36 interacts with HCV E1 and might be a co-receptor specific for HCV entry; thus, CD36 could be a potential drug target against HCV. PMID:26898231

  9. Harmonic two-phonon {gamma}-vibrational state in neutron-rich {sup 106}Mo

    SciTech Connect

    Guessous, A.; Schulz, N.; Phillips, W.R.; Ahmad, I.; Bentaleb, M.; Durell, J.L.; Jones, M.A.; Leddy, M.; Lubkiewicz, E.; Morss, L.R.; Piepenbring, R.; Smith, A.G.; Urban, W.; Varley, B.J.

    1995-09-18

    The neutron-rich {sup 106}Mo nucleus has been studied by prompt {gamma}-ray spectroscopy following the spontaneous fission of {sup 248}Cm. The characteristics of the rotational band built on a state at 1435 keV identifies the band head as the best candidate for a harmonic double-phonon {ital K}{sup {pi}}=4{sup +} {gamma}-vibrational state observed so far.

  10. 26 CFR 1.72(e)-1T - Treatment of distributions where substantially all contributions are employee contributions...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 2 2014-04-01 2014-04-01 false Treatment of distributions where substantially all contributions are employee contributions (temporary). 1.72(e)-1T Section 1.72(e)-1T Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Items Specifically Included...

  11. Cytochrome P4502E1 inhibitor, chlormethiazole, decreases lipopolysaccharide-induced inflammation in rat Kupffer cells with ethanol treatment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    To investigate the role of Cytochrome P4502E1 in sensitizing Kupffer cells to lipopolysaccharide (LPS)-mediated inflammation after ethanol induction. Sprague-Dawley rats were fed a liquid ethanol diet, control diet or ethanol diet supplemented with CYP2E1 inhibitor, chlormethiazole (CMZ), for 4'week...

  12. Partition of E1A proteins between soluble and structural fractions of adenovirus-infected and -transformed cells.

    PubMed Central

    Chatterjee, P K; Flint, S J

    1986-01-01

    The partition of E1A proteins between soluble and structural framework fractions of human cells infected or transformed by subgroup C adenoviruses was investigated by using gentle cell fractionation conditions. A polyclonal antibody raised against a trpE-E1A fusion protein (K.R. Spindler, D.S.E. Rosser, and A. J. Berk, J. Virol. 132-141, 1984) synthesized in Escherichia coli was used to measure the steady-state levels of E1A proteins recovered in the various fractions by immunoblotting. The relative concentration of E1A proteins recovered in the soluble fraction of adenovirus type 2-infected cells was at least fivefold greater than the relative concentration in the corresponding fraction of transformed 293 cells. The observed distribution of E1A proteins was not altered by the sulfhydryl-blocking reagent N-ethylmaleimide. E1A proteins were recovered in nuclear matrix, chromatin, and cytoskeleton fractions after further fractionation of the structural framework fraction. However, the E1A protein species that could be identified by one-dimensional gel electrophoresis were not uniformly distributed among the subcellular fractions examined. The results obtained when fractionation was performed in the presence of the oxidation catalysts Cu2+ or (ortho-phenanthroline)2 Cu2+ indicate that E1A proteins can be efficiently cross-linked, via disulfide bonds, to the structural framework of both adenovirus-infected and adenovirus-transformed cells. Images PMID:3023654

  13. Complete Genome Sequence of Methanosphaerula palustris E1-9CT, a Hydrogenotrophic Methanogen Isolated from a Minerotrophic Fen Peatland

    PubMed Central

    Browne, Patrick; Kyrpides, Nikos; Woyke, Tanja; Goodwin, Lynne; Detter, Chris; Yavitt, Joseph B.; Zinder, Stephen H.

    2015-01-01

    Here, we report the complete genome sequence (2.92Mb) of Methanosphaerula palustris E1-9CT, a methanogen isolated from a minerotrophic fen. This is the first genome report of the Methanosphaerula genus, within the Methanoregulaceae family, in the Methanomicrobiales order. E1-9CT relatives are found in a wide range of ecological and geographical settings. PMID:26543115

  14. 26 CFR 1.1033(e)-1 - Sale or exchange of livestock solely on account of drought.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... of drought. 1.1033(e)-1 Section 1.1033(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF... Sale or exchange of livestock solely on account of drought. (a) The sale or exchange of livestock... if the sale or exchange of such livestock by the taxpayer is solely on account of drought....

  15. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties

    PubMed Central

    Beaumont, Elodie; Roch, Emmanuelle; Chopin, Lucie; Roingeard, Philippe

    2016-01-01

    Various strategies involving the use of hepatitis C virus (HCV) E1 and E2 envelope glycoproteins as immunogens have been developed for prophylactic vaccination against HCV. However, the ideal mode of processing and presenting these immunogens for effective vaccination has yet to be determined. We used our recently described vaccine candidate based on full-length HCV E1 or E2 glycoproteins fused to the heterologous hepatitis B virus S envelope protein to compare the use of the E1 and E2 proteins as separate immunogens with their use as the E1E2 heterodimer, in terms of immunogenetic potential and the capacity to induce neutralizing antibodies. The specific anti-E1 and anti-E2 antibody responses induced in animals immunized with vaccine particles harboring the heterodimer were profoundly impaired with respect to those in animals immunized with particles harboring E1 and E2 separately. Moreover, the anti-E1 and anti-E2 antibodies had additive neutralizing properties that increase the cross-neutralization of heterologous strains of various HCV genotypes, highlighting the importance of including both E1 and E2 in the vaccine for an effective vaccination strategy. Our study has important implications for the optimization of HCV vaccination strategies based on HCV envelope proteins, regardless of the platform used to present these proteins to the immune system. PMID:26966906

  16. Functional analysis of the C-terminal region of human adenovirus E1A reveals a misidentified nuclear localization signal

    SciTech Connect

    Cohen, Michael J.; King, Cason R.; Dikeakos, Jimmy D.; Mymryk, Joe S.

    2014-11-15

    The immortalizing function of the human adenovirus 5 E1A oncoprotein requires efficient localization to the nucleus. In 1987, a consensus monopartite nuclear localization sequence (NLS) was identified at the C-terminus of E1A. Since that time, various experiments have suggested that other regions of E1A influence nuclear import. In addition, a novel bipartite NLS was recently predicted at the C-terminal region of E1A in silico. In this study, we used immunofluorescence microscopy and co-immunoprecipitation analysis with importin-α to verify that full nuclear localization of E1A requires the well characterized NLS spanning residues 285–289, as well as a second basic patch situated between residues 258 and 263 ({sup 258}RVGGRRQAVECIEDLLNEPGQPLDLSCKRPRP{sup 289}). Thus, the originally described NLS located at the C-terminus of E1A is actually a bipartite signal, which had been misidentified in the existing literature as a monopartite signal, altering our understanding of one of the oldest documented NLSs. - Highlights: • Human adenovirus E1A is localized to the nucleus. • The C-terminus of E1A contains a bipartite nuclear localization signal (NLS). • This signal was previously misidentified to be a monopartite NLS. • Key basic amino acid residues within this sequence are highly conserved.

  17. RORα switches transcriptional mode of ERRγ that results in transcriptional repression of CYP2E1 under ethanol-exposure

    PubMed Central

    Han, Yong-Hyun; Kim, Don-Kyu; Na, Tae-Young; Ka, Na-Lee; Choi, Hueng-Sik; Lee, Mi-Ock

    2016-01-01

    Increased cytochrome P450 2E1 (CYP2E1) expression is the main cause of oxidative stress, which exacerbates alcoholic liver diseases (ALDs). Estrogen-related receptor gamma (ERRγ) induces CYP2E1 expression and contributes to enhancing alcohol-induced liver injury. Retinoic acid-related orphan receptor alpha (RORα) has antioxidative functions; however, potential cross-talk between ERRγ and RORα in the regulation of CYP2E1 has not been studied. We report that RORα suppressed ERRγ-mediated CYP2E1 expression. A physical interaction of RORα with ERRγ at the ERRγ−response element in the CYP2E1 promoter was critical in this suppression. At this site, coregulator recruitment of ERRγ was switched from coactivator p300 to the nuclear receptor corepressor 1 in the presence of RORα. Cross-talk between ERRγ and RORα was demonstrated in vivo, in that administration of JC1–40, a RORα activator, significantly decreased both CYP2E1 expression and the signs of liver injury in ethanol-fed mice, and this was accompanied by coregulator switching. Thus, this non-classical RORα pathway switched the transcriptional mode of ERRγ, leading to repression of alcohol-induced CYP2E1 expression, and this finding may provide a new therapeutic strategy against ALDs. PMID:26464440

  18. Extra γ-ray strength for 116,117Sn arising from pygmy dipole resonance

    NASA Astrophysics Data System (ADS)

    Kamata, M.; Utsunomiya, H.; Akimune, H.; Yamagata, T.; Itoh, O.; Iwamoto, C.; Kondo, T.; Toyokawa, H.; Lui, Y.-W.; Goriely, S.

    2010-06-01

    Photoneutron cross sections were measured for 117Sn and 116Sn near neutron thresholds with quasi-monochromatic laser Compton scattering γ-rays. The measured cross sections for 117Sn and 116Sn are strongly enhanced from the threshold behavior expected for L = 1 neutron emissions after E1 photoexcitation. This suggests the presence of extra γ-ray strength in the low-energy tail of the giant dipole resonance. The present cross sections were analyzed together with radiative neutron capture cross sections for 116Sn within the framework of the statistical model calculation. It is shown that the extra γ-ray strength can be interpreted as pygmy E1 resonance which was previously reported in the nuclear resonance fluorescence experiment for 116Sn and 124Sn.

  19. Characterization and Whole Genome Analysis of Human Papillomavirus Type 16 E1-1374∧63nt Variants

    PubMed Central

    Sabol, Ivan; Matovina, Mihaela; Si-Mohamed, Ali; Grce, Magdalena

    2012-01-01

    Background The variation of the most common Human papillomavirus (HPV) type found in cervical cancer, the HPV16, has been extensively investigated in almost all viral genes. The E1 gene variation, however, has been rarely studied. The main objective of the present investigation was to analyze the variability of the E6 and E1 genes, focusing on the recently identified E1-1374∧63nt variant. Methodology/Principal Findings Variation within the E6 of 786 HPV16 positive cervical samples was analyzed using high-resolution melting, while the E1-1374∧63nt duplication was assayed by PCR. Both techniques were supplemented with sequencing. The E1-1374∧63nt duplication was linked with the E-G350 and the E-C109/G350 variants. In comparison to the referent HPV16, the E1-1374∧63nt E-G350 variant was significantly associated with lower grade cervical lesions (p = 0.029), while the E1-1374∧63nt E-C109/G350 variant was equally distributed between high and low grade lesions. The E1-1374∧63nt variants were phylogenetically closest to E-G350 variant lineage (A2 sub-lineage based on full genome classification). The major differences between E1-1374∧63nt variants were within the LCR and the E6 region. On the other hand, changes within the E1 region were the major differences from the A2 sub-lineage, which has been historically but inconclusively associated with high grade cervical disease. Thus, the shared variations cannot explain the particular association of the E1-1374∧63nt variant with lower grade cervical lesions. Conclusions/Significance The E1 region has been thus far considered to be well conserved among all HPVs and therefore uninteresting for variability studies. However, this study shows that the variations within the E1 region could possibly affect cervical disease, since the E1-1374∧63nt E-G350 variant is significantly associated with lower grade cervical lesions, in comparison to the A1 and A2 sub-lineage variants. Furthermore, it appears that the silent variation 109T>C of the E-C109/G350 variant might have a significant role in the viral life cycle and warrants further study. PMID:22911739

  20. Radiative rates for E1, E2, M1, and M2 transitions in F-like ions with 37 ≤ Z ≤ 53

    NASA Astrophysics Data System (ADS)

    Aggarwal, Kanti M.; Keenan, Francis P.

    2016-05-01

    Calculations of energy levels, radiative rates and lifetimes are reported for 17 F-like ions with 37≤Z≤53. For brevity, results are only presented among the lowest 113 levels of the 2s22p5, 2s2p6, 2s22p43 ℓ, 2s2p53 ℓ, and 2p63 ℓ configurations, although the calculations have been performed for up to 501 levels in each ion. The general-purpose relativistic atomic structure package (GRASP) has been adopted for the calculations, and radiative rates (along with oscillator strengths and line strengths) are listed for all E1, E2, M1, and M2 transitions of the ions. Comparisons are made with earlier available experimental and theoretical energies, although these are limited to only a few levels for most ions. Therefore for additional accuracy assessments, particularly for energy levels, analogous calculations have been performed with the Flexible Atomic Code (FAC), for up to 72 259 levels. Limited previous results are available for radiative rates for comparison purposes, and no large discrepancy is observed for any transition and/or ion.

  1. Strength Scaling in Fiber Composites

    NASA Technical Reports Server (NTRS)

    Kellas, Sotiris; Morton, John

    1990-01-01

    A research program was initiated to study and isolate the factors responsible for scale effects in the tensile strength of graphite/epoxy composite laminates. Four layups were chosen with appropriate stacking sequences so as to highlight individual and interacting failure modes. Four scale sizes were selected for investigation including full scale size, 3/4, 2/4, and 1/4, with n = to 4, 3, 2, and 1, respectively. The full scale specimen sizes was 32 piles thick as compared to 24, 16, and 8 piles for the 3/4, 2/4, and 1/4 specimen sizes respectively. Results were obtained in the form of tensile strength, stress-strain curves and damage development. Problems associated with strength degradation with increasing specimen sizes are isolated and discussed. Inconsistencies associated with strain measurements were also identified. Enhanced x ray radiography was employed for damage evaluation, following step loading. It was shown that fiber dominated layups were less sensitive to scaling effects compared to the matrix dominated layups.

  2. An Interlaminar Tensile Strength Specimen

    NASA Technical Reports Server (NTRS)

    Martin, Roderick H.; Jackson, Wade C.

    1993-01-01

    This paper describes a technique to determine interlaminar tensile strength, sigma(sub 3c), of a fiber reinforced composite material using a curved beam. The specimen was a unidirectional curved beam, bent 90 deg, with straight arms. Attached to each arm was a hinged loading mechanism that was held by the grips of a tension testing machine. Geometry effects of the specimen, including the effects of loading arm length, inner radius, thickness, and width, were studied. The data sets fell into two categories: low strength corresponding to a macroscopic flaw related failure and high strength corresponding to a microscopic flaw related failure. From the data available, the specimen width and loading arm length had little effect on sigma(sub 3c). The inner radius was not expected to have a significant effect on sigma(sub 3c), but this conclusion could not be confirmed because of differences in laminate quality for each curve geometry. The thicker specimens had the lowest value of sigma(sub 3c) because of poor laminate quality.

  3. Shear Strength of Aluminum Oxynitride

    NASA Astrophysics Data System (ADS)

    Dandekar, Dattatraya P.; Vaughan, Brian A. M.; Proud, William G.

    2007-06-01

    Aluminum oxynitride (AlON) is a transparent, polycrystalline cubic spinel. The results of investigations^1-4 on shock response of AlON permit determination of the equation of state, and shear strength retained under shock compression. Whereas the values of the HEL of AlON holds no surprises, the inelastic response of AlON reported in Ref. 1-4 differ significantly and is stress dependent. The results of Ref. 1-2 show that AlON retains a shear strength of 3 to 4 GPa when shocked up to around 20 GPa, but the results of Ref, 3-4 seem to suggest a possible loss of shear strength when shocked to 16 GPa and beyond. Our analysis examines the observed differences in the inelastic response of AlON reported in these four studies . 1. J. U. Cazamias, et. al., in Fundamental Issues and Applications of Shock-Wave and High Strain Rate Phenomena, Eds. Staudhammer, Murr, and Meyers, Elsevier, NY, 173 (2001). 2. B. A. M. Vaughn, et.al., Shock Physics, Cavendish Laboratory, Report SP/1092 (2001) 3. T. Sekine, et.al., J. Appl. Phys. 94, 4803 (2003). 4. T. F. Thornhill, et.al., Shock Compression of Matter-2005, Eds. Furnish, Elert, Russell, White, AIP, NY, 143 (2006).

  4. The Inactivation of Human CYP2E1 by Phenethyl Isothiocyanate, a Naturally Occurring Chemopreventive Agent, and Its Oxidative Bioactivation

    PubMed Central

    Yoshigae, Yasushi; Sridar, Chitra; Kent, Ute M.

    2013-01-01

    Phenethylisothiocyanate (PEITC), a naturally occurring isothiocyanate and potent cancer chemopreventive agent, works by multiple mechanisms, including the inhibition of cytochrome P450 (P450) enzymes, such as CYP2E1, that are involved in the bioactivation of carcinogens. PEITC has been reported to be a mechanism-based inactivator of some P450s. We describe here the possible mechanism for the inactivation of human CYP2E1 by PEITC, as well as the putative intermediate that might be involved in the bioactivation of PEITC. PEITC inactivated recombinant CYP2E1 with a partition ratio of 12, and the inactivation was not inhibited in the presence of glutathione (GSH) and not fully recovered by dialysis. The inactivation of CYP2E1 by PEITC is due to both heme destruction and protein modification, with the latter being the major pathway for inactivation. GSH-adducts of phenethyl isocyanate (PIC) and phenethylamine were detected during the metabolism by CYP2E1, indicating formation of PIC as a reactive intermediate following P450-catalyzed desulfurization of PEITC. Surprisingly, PIC bound covalently to CYP2E1 to form protein adducts but did not inactivate the enzyme. Liquid chromatography mass spectroscopy analysis of the inactivated CYP2E1 apo-protein suggests that a reactive sulfur atom generated during desulfurization of PEITC is involved in the inactivation of CYP2E1. Our data suggest that the metabolism of PEITC by CYP2E1 that results in the inactivation of CYP2E1 may occur by a mechanism similar to that observed with other sulfur-containing compounds, such as parathion. Digestion of the inactivated enzyme and analysis by SEQUEST showed that Cys 268 may be the residue modified by PIC. PMID:23371965

  5. MicroRNA-16 Modulates HuR Regulation of Cyclin E1 in Breast Cancer Cells

    PubMed Central

    Guo, Xun; Connick, Melanie C.; Vanderhoof, Jennifer; Ishak, Mohammad-Ali; Hartley, Rebecca S.

    2015-01-01

    RNA binding protein (RBPs) and microRNAs (miRNAs or miRs) are post-transcriptional regulators of gene expression that are implicated in development of cancers. Although their individual roles have been studied, the crosstalk between RBPs and miRNAs is under intense investigation. Here, we show that in breast cancer cells, cyclin E1 upregulation by the RBP HuR is through specific binding to regions in the cyclin E1 mRNA 3' untranslated region (3'UTR) containing U-rich elements. Similarly, miR-16 represses cyclin E1, dependent on its cognate binding sites in the cyclin E1 3'UTR. Evidence in the literature indicates that HuR can regulate miRNA expression and recruit or dissociate RNA-induced silencing complexes (RISC). Despite this, miR-16 and HuR do not affect the other’s expression level or binding to the cyclin E1 3'UTR. While HuR overexpression partially blocks miR-16 repression of a reporter mRNA containing the cyclin E1 3'UTR, it does not block miR-16 repression of endogenous cyclin E1 mRNA. In contrast, miR-16 blocks HuR-mediated upregulation of cyclin E1. Overall our results suggest that miR-16 can override HuR upregulation of cyclin E1 without affecting HuR expression or association with the cyclin E1 mRNA. PMID:25830480

  6. Phylogeography of E1b1b1b-M81 haplogroup and analysis of its subclades in Morocco.

    PubMed

    Reguig, Ahmed; Harich, Nourdin; Barakat, Abdelhamid; Rouba, Hassan

    2014-01-01

    In this study we analyzed 295 unrelated Berber-speaking men from northern, central, and southern Morocco to characterize frequency of the E1b1b1b-M81 haplogroup and to refine the phylogeny of its subclades: E1b1b1b1-M107, E1b1b1b2-M183, and E1b1b1b2a-M165. For this purpose, we typed four biallelic polymorphisms: M81, M107, M183, and M165. A large majority of the Berber-speaking male lineages belonged to the Y-chromosomal E1b1b1b-M81 haplogroup. The frequency ranged from 79.1% to 98.5% in all localities sampled. E1b1b1b2-M183 was the most dominant subclade in our samples, ranging from 65.1% to 83.1%. In contrast, the E1b1b1b1-M107 and E1b1b1b2a-M165 subclades were not found in our samples. Our results suggest a predominance of the E1b1b1b-M81 haplogroup among Moroccan Berber-speaking males with a decreasing gradient from south to north. The most prevalent subclade in this haplogroup was E1b1b1b2-M183, for which diffferences among these three groups were statistically significant between central and southern groups. PMID:25397701

  7. Strength loss in kraft pulping

    NASA Astrophysics Data System (ADS)

    Iribarne, Jose

    Unbleached kraft pulps from two U.S. mills were 21% and 26% weaker than comparable laboratory pulps from the same chip sources, when assessed as the tear index at a tensile index of 70 kN.m/kg. The phenomena involved were clarified by characterizing the differences between the mill and laboratory pulps in terms of fundamental fiber properties. All of the strength loss could be explained by a reduction in intrinsic fiber strength of 9% to 11%, as estimated from wet zero-span tensile tests and fiber length distributions. Most of the effects of different fiber shape and length were isolated by PFI mill refining and decrilling, respectively. The higher fiber coarseness of mill pulps was a factor in their maximum density and bond strength, but changes in these variables were analogous to those of laboratory pups due to similar swelling. Specific bond strength, determined from a wet pressing experiment, was similar in mill and laboratory pulps. Neither carbohydrate composition nor crystalline structure, assessed through x-ray diffraction analysis, were significant factors in the observed fiber strength differences. The mill pulps were not more heterogeneous than the laboratory pulps, within the resolution of a fractionation experiment. The number of weak points in each pulp was assessed through analysis of the amount of fiber cutting during PFI mill refining and treatments with potassium superoxide or cellulase. The results suggested that the chemistry of kraft pulping preferentially weaken short, slender fibers, while mechanical stresses during the hot discharge of batch digesters mainly affect long, thick fibers. The greater number of weak points in the long-fiber fractions of mill pulps is probably associated with their lower wet zero-span tensile indices. Automated optical detection of major singularities with a prototype instrument suggested that only the weak points induced by mechanical stress could be detected by local variations in birefringence. In contrast, chemically damaged short, slender fibers were not optically active. Strong chemical attack during superoxide treatment appeared to affect all fibers, but the effect of fiber cutting was partially offset by a preferential dissolution of short fibers and fines. A simple model of weak point formation by combinations of mechanical stress and localized chemical attack was sufficient to explain all the experimental results.

  8. Heterogeneity of adenovirus type 5 E1A proteins: multiple serine phosphorylations induce slow-migrating electrophoretic variants but do not affect E1A-induced transcriptional activation or transformation.

    PubMed Central

    Richter, J D; Slavicek, J M; Schneider, J F; Jones, N C

    1988-01-01

    The 289-amino-acid product encoded by the adenovirus E1A 13S mRNA has several pleiotropic activities, including transcriptional activation, transcriptional repression, and when acting in concert with certain oncogene products, cell transformation. In all cell types in which E1A has been introduced (except bacteria), E1A protein is extensively posttranslationally modified to yield several isoelectric and molecular weight variants. The most striking variant is one that has a retarded mobility, by about Mr = 2,000, in sodium dodecyl sulfate gels. We have investigated the nature of this modification and have assessed its importance for E1A activity. Phosphorylation is responsible for the altered mobility of E1A, since acid phosphatase treatment eliminates the higher apparent molecular weight products. By using several E1A deletion mutants, we show that at least two seryl residues, residing between residues 86 and 120 and 224 and 289, are the sites of phosphorylation and that each phosphorylation can independently induce the mobility shift. However, E1A mutants lacking these seryl residues transcriptionally activate the adenovirus E3 and E2A promoters and transform baby rat kidney cells to near wild-type levels. Images PMID:2835499

  9. The Measurements of Gamma-ray Spectrometers from China's Chang'E-1/2 Spacecrafts

    NASA Astrophysics Data System (ADS)

    Zhu, Meng-Hua; Chang, Jin; Ma, T.

    2012-07-01

    Gamma ray spectrometers, as one of the major payloads onboard China's Chang'E-1/2 spacecrafts, were aimed to provide maps of the abundances of major elements, O, Si, Mg, Al, Ca, Ti, Na, and Fe, and of the natural radioactive elements, U, Th, and K, in the subsurface of the Moon. These elements presented on the lunar surface are the end products of a series of processes (i.e., accumulation, collision, and modification), their measurements, therefore, could provide many important clues to understand the formation and evolution of the Moon. Gamma ray spectrometer onboard Chang'E-1spacecraft used a large CsI(Tl) crystal as its main detector to measure gamma rays from the Moon with energy resolution of ˜9%@662 keV while Chang'E-2 gamma ray spectrometer firstly used big LaBr3 crystal in the planetary detection with energy resolution of ˜3.61%@662 keV. Both gamma ray spectrometers used coincident technique to suppress the Compton effects and reduce the background gamma rays from the interactions from GCRs with spacecraft materials. Gamma rays from the Moon were measured in the energy range of 0.3 ˜ 10 MeV with 512-channel spectrum every 3 seconds from a circle, polar-orbit but at a nominal altitude of 200 km and 100 km, respectively. Thorium and Potassium maps were derived from the one-year gamma ray measurements of Chang'E-1spacecraft. All these maps show good consistent with previous results from Lunar Prospector and Kaguya missions. Lunar terrains could be clearly classified on the basis of these compositions in relation with other characteristics. Chang'E-2 gamma ray spectrometer has only half-year detections, but since large LaBr3 crystal has higher energy resolution and higher detection efficiency, element distribution across the lunar surface could be obtained with higher precision. With the data set from Chang'E-2 gamma ray spectrometer, radioactive elements (K, and Th), and major elements (Fe, Ti, Si, Mg, Al, O, and Ca) distributions on the lunar surface have been derived. From present research, several new features have been found that are different from the results of Lunar Prospector gamma ray spectrometer.

  10. Molecular basis of maple syrup urine disease: Novel mutations at the E1[alpha] locus that impair E1([alpha][sub 2][beta][sub 2]) assembly or decrease steady-state E1[alpha] mRNA levels of branched-chain [alpha]-keto acid dehydrogenase complex

    SciTech Connect

    Chuang, J.L.; Fisher, C.R.; Chuang, D.T.; Cox, R.P. )

    1994-08-01

    The authors report the occurrence of three novel mutations in the E1[alpha] (BCKDHA) locus of the branched-chain [alpha]-keto acid dehydrogenase (BCKAD) complex that cause maple syrup urine disease (MSUD). An 8-bp deletion in exon 7 is present in one allele of a compound-heterozygous patient (GM-649). A single C nucleotide insertion in exon 2 occurs in one allele of an intermediate-MSUD patient (Lo). The second allele of patient Lo carries an A-to-G transition in exon 9 of the E1[alpha] gene. This missense mutation changes Tyr-368 to Cys (Y368C) in the E1[alpha] subunit. Both the 8-bp deletion and the single C insertion generate a downstream nonsense codon. Both mutations appear to be associated with a low abundance of the mutant E1[alpha] mRNA, as determined by allele-specific oligonucleotide probing. Transfection studies strongly suggest that the Y368C substitution in the E1[alpha] subunit impairs its proper assembly with the normal E1[beta]. Unassembled as well as misassembled E1[alpha] and E1[beta] subunits are degraded in the cell. 32 refs., 8 figs.

  11. HBx inhibits CYP2E1 gene expression via downregulating HNF4α in human hepatoma cells.

    PubMed

    Liu, Hongming; Lou, Guiyu; Li, Chongyi; Wang, Xiaodong; Cederbaum, Arthur I; Gan, Lixia; Xie, Bin

    2014-01-01

    CYP2E1, one of the cytochrome P450 mixed-function oxidases located predominantly in liver, plays a key role in metabolism of xenobiotics including ethanol and procarcinogens. Recently, down-expression of CYP2E1 was found in hepatocellular carcinoma (HCC) with the majority to be chronic hepatitis B virus (HBV) carriers. In this study, we tested a hypothesis that HBx may inhibit CYP2E1 gene expression via hepatocyte nuclear factor 4α (HNF4α). By enforced HBx gene expression in cultured HepG2 cells, we determined the effect of HBx on CYP2E1 mRNA and protein expression. With a bioinformatics analysis, we found a consensus HNF-4α binding sequence located on -318 to -294 bp upstream of human CYP2E1 promoter. Using reporter gene assay and site-directed mutagenesis, we have shown that mutation of this site dramatically decreased CYP2E1 promoter activity. By silencing endogenous HNF-4α, we have further validated knockdown of HNF-4α significantly decreased CYP2E1 expression. Ectopic overexpression of HBx in HepG2 cells inhibits HNF-4α expression, and HNF-4α levels were inversely correlated with viral proteins both in HBV-infected HepG2215 cells and as well as HBV positive HCC liver tissues. Moreover, the HBx-induced CYP2E1 reduction could be rescued by ectopic supplement of HNF4α protein expression. Furthermore, human hepatoma cells C34, which do not express CYP2E1, shows enhanced cell growth rate compared to E47, which constitutively expresses CYP2E1. In addition, the significantly altered liver proteins in CYP2E1 knockout mice were detected with proteomics analysis. Together, HBx inhibits human CYP2E1 gene expression via downregulating HNF4α which contributes to promotion of human hepatoma cell growth. The elucidation of a HBx-HNF4α-CYP2E1 pathway provides novel insight into the molecular mechanism underlining chronic HBV infection associated hepatocarcinogenesis. PMID:25238230

  12. Ion selectivity of colicin E1: II. Permeability to organic cations.

    PubMed

    Bullock, J O; Kolen, E R; Shear, J L

    1992-05-01

    Channels formed by colicin E1 in planar lipid bilayers have large diameters and conduct both cations and anions. The rates at which ions are transported, however, are relatively slow, and the relative anion-to-cation selectivity is modulated over a wide range by the pH of the bathing solutions. We have examined the permeability of these channels to cationic probes having a variety of sizes, shapes, and charge distributions. All of the monovalent probes were found to be permeant, establishing a minimum diameter at the narrowest part of the pore of approximately 9 A. In contrast to this behavior, all of the polyvalent organic cations were shown to be impermeant. This simple exclusionary rule is interpreted as evidence that, when steric restrictions require partial dehydration of an ion, the structure of the channel is able to provide a substitute electrostatic environment for only one charged group at time. PMID:1380089

  13. Combined System of Activated Sludge and Ozonation for the Treatment of Kraft E1 Effluent

    PubMed Central

    Assalin, Marcia Regina; dos Santos Almeida, Edna; Durán, Nelson

    2009-01-01

    The treatment of paper mill effluent for COD, TOC, total phenols and color removal was investigated using combined activated sludge-ozonation processes and single processes. The combined activated sludge-O3/pH 10 treatment was able to remove around 80% of COD, TOC and color from Kraft E1 effluent. For the total phenols, the efficiency removal was around 70%. The ozonation post treatment carried out at pH 8.3 also showed better results than the single process. The COD, TOC, color and total phenols removal efficiency obtained were 75.5, 59.1, 77 and 52.3%, respectively. The difference in the concentrations of free radical produced by activated sludge-O3/pH 10 and activated sludge-O3/pH 8.3 affected mainly the TOC and total phenol removal values. PMID:19440438

  14. Characterization of a nitrophenol reductase from the phototrophic bacterium Rhodobacter capsulatus E1F1.

    PubMed Central

    Blasco, R; Castillo, F

    1993-01-01

    The phototrophic bacterium Rhodobacter capsulatus E1F1 photoreduced 2,4-dinitrophenol to 2-amino-4-nitrophenol by a nitrophenol reductase activity which was induced in the presence of nitrophenols and was repressed in ammonium-grown cells. The enzyme was located in the cytosol, required NAD(P)H as an electron donor, and used several nitrophenol derivatives as alternative substrates. The nitrophenol reductase was purified to electrophoretic homogeneity by a simple method. The enzyme was composed of two 27-kDa subunits, was inhibited by metal chelators, mercurial compounds, and Cu2+, and contained flavin mononucleotide and possibly nonheme iron as prosthetic groups. Purified enzyme also exhibited NAD(P)H diaphorase activity which used tetrazolium salt as an electron acceptor. Images PMID:8328801

  15. Health sector response to security threats during the civil war in E1 Salvador.

    PubMed Central

    Brentlinger, P. E.

    1996-01-01

    During the recent civil war in E1 Salvador, as in other modern wars, human rights abuses adversely affected health workers, patients, and medical facilities. The abuses themselves have been described in reports of human rights advocacy organisations but health sector adaptations to a hostile wartime environment have not. Agencies engaged in health work during the civil war adapted parties such as training of community based lay health workers, use of simple technology, concealment of patients and medical supplies, denunciation of human rights abuses, and multilevel negotiations in order to continue providing services. The Salvadorean experience may serve as a helpful case study for medical personnel working in wars elsewhere. Images p1471-a p1472-a p1473-a PMID:8973238

  16. The effect of prostaglandin E1 on motility of the equine gut.

    PubMed

    Hunt, J M; Gerring, E L

    1985-06-01

    Prostaglandin E1 was infused intravenously (25, 50 and 75 ng/kg/min) in three ponies. Changes in gastrointestinal mechanical and electrical activity were recorded from chronically implanted strain-gauge force transducers and electrodes. Dose-dependent responses were obtained: there were significant decreases in electrical spiking activity in the stomach, left large colon and small colon, with a corresponding decrease of activity in the left dorsal colon mechanogram. The small intestine was also affected, showing a decrease in both contraction rate and amplitude, which was more marked in the proximal jejunum than in the ileum. There was an association between these changes in gastrointestinal activity and the presence of discomfort and diminished gut sounds. PMID:4040579

  17. Inert strength of pristine silica glass fibers

    SciTech Connect

    Smith, W.L.; Michalske, T.A.

    1993-11-01

    Silica glass fibers have been produced and tested under ultra high vacuum (UHV) conditions to investigate the inert strength of pristine fibers in absence of reactive agents. Analysis of the coefficient of variation in diameter ({upsilon}{sub d}) vs the coefficient of variation of breaking strength ({upsilon}{sub {sigma}}) does not adequately explain the variation of breaking stress. Distribution of fiber tensile strength data suggests that the inert strength of such fibers is not single valued and that the intrinsic strength is controlled by defects in the glass. Furthermore, comparison of room temperature UHV data with LN{sub 2} data indicates that these intrinsic strengths are not temperature dependent.

  18. Melatonin Suppresses Autophagy Induced by Clinostat in Preosteoblast MC3T3-E1 Cells.

    PubMed

    Yoo, Yeong-Min; Han, Tae-Young; Kim, Han Sung

    2016-01-01

    Microgravity exposure can cause cardiovascular and immune disorders, muscle atrophy, osteoporosis, and loss of blood and plasma volume. A clinostat device is an effective ground-based tool for simulating microgravity. This study investigated how melatonin suppresses autophagy caused by simulated microgravity in preosteoblast MC3T3-E1 cells. In preosteoblast MC3T3-E1 cells, clinostat rotation induced a significant time-dependent increase in the levels of the autophagosomal marker microtubule-associated protein light chain (LC3), suggesting that autophagy is induced by clinostat rotation in these cells. Melatonin treatment (100, 200 nM) significantly attenuated the clinostat-induced increases in LC3 II protein, and immunofluorescence staining revealed decreased levels of both LC3 and lysosomal-associated membrane protein 2 (Lamp2), indicating a decrease in autophagosomes. The levels of phosphorylation of mammalian target of rapamycin (p-mTOR) (Ser2448), phosphorylation of extracellular signal-regulated kinase (p-ERK), and phosphorylation of serine-threonine protein kinase (p-Akt) (Ser473) were significantly reduced by clinostat rotation. However, their expression levels were significantly recovered by melatonin treatment. Also, expression of the Bcl-2, truncated Bid, Cu/Zn- superoxide dismutase (SOD), and Mn-SOD proteins were significantly increased by melatonin treatment, whereas levels of Bax and catalase were decreased. The endoplasmic reticulum (ER) stress marker GRP78/BiP, IRE1α, and p-PERK proteins were significantly reduced by melatonin treatment. Treatment with the competitive melatonin receptor antagonist luzindole blocked melatonin-induced decreases in LC3 II levels. These results demonstrate that melatonin suppresses clinostat-induced autophagy through increasing the phosphorylation of the ERK/Akt/mTOR proteins. Consequently, melatonin appears to be a potential therapeutic agent for regulating microgravity-related bone loss or osteoporosis. PMID:27070587

  19. Joint Tracking of Chang'E-1 with VLBI and USB

    NASA Astrophysics Data System (ADS)

    Hu, Xiaogong

    Chinese lunar exploration mission Chang'E-I made use of a Chinese Unified S-Band (USB) system and a network of four Very Long Baseline Interferometry (VLBI) antennas to meet the orbit determination/predication requirements of spacecraft tracking and scientific data analysis, which for the first time handled telemetry and control for a spacecraft at a distance of about 380,000 km. Chang'E-1 provided a perfect chance to quantify the contributions of VLBI to orbit determination/predication, and to test and evaluate the performance of the USB-VLBI joint system. We investigate in this paper the quality of the data and analyze the precision of orbit determination with different data arcs and data combinations during the 2-week journey from Earth to Chang'E-1's lunar mission orbit, using GEODYN II orbit determination software. The residuals of VLBI delay is about 3 ns (RMS, root-mean-squares), the residuals of VLBI delay-rate is about 0.6 ps/s, the residuals of ranging is about 1 2 m and Doppler is about 1 cm/s. Three flight phases are invistigated, namely 3 phasing orbits near Earth, the translunar trajectory and the lunar catputed orbits. We found including of VLBI data substantially improved orbit precision for short data arcs, therefore VLBI played an important role in assessing spacecraft manuvore performance. Given the differential nature of VLBI observables and their errors mostly from BBC's nonlinear phase-frequency responses, it appears for longer data arcs the VLBI contribution is relatively minor. We conclude that for Chang'E-I, the inclusion of VLBI data improved substantially the performance of orbit determination and prediction, even though the VLBI data acquisition and correlation imposes a major burden on data processing.

  20. Melatonin Suppresses Autophagy Induced by Clinostat in Preosteoblast MC3T3-E1 Cells

    PubMed Central

    Yoo, Yeong-Min; Han, Tae-Young; Kim, Han Sung

    2016-01-01

    Microgravity exposure can cause cardiovascular and immune disorders, muscle atrophy, osteoporosis, and loss of blood and plasma volume. A clinostat device is an effective ground-based tool for simulating microgravity. This study investigated how melatonin suppresses autophagy caused by simulated microgravity in preosteoblast MC3T3-E1 cells. In preosteoblast MC3T3-E1 cells, clinostat rotation induced a significant time-dependent increase in the levels of the autophagosomal marker microtubule-associated protein light chain (LC3), suggesting that autophagy is induced by clinostat rotation in these cells. Melatonin treatment (100, 200 nM) significantly attenuated the clinostat-induced increases in LC3 II protein, and immunofluorescence staining revealed decreased levels of both LC3 and lysosomal-associated membrane protein 2 (Lamp2), indicating a decrease in autophagosomes. The levels of phosphorylation of mammalian target of rapamycin (p-mTOR) (Ser2448), phosphorylation of extracellular signal-regulated kinase (p-ERK), and phosphorylation of serine-threonine protein kinase (p-Akt) (Ser473) were significantly reduced by clinostat rotation. However, their expression levels were significantly recovered by melatonin treatment. Also, expression of the Bcl-2, truncated Bid, Cu/Zn- superoxide dismutase (SOD), and Mn-SOD proteins were significantly increased by melatonin treatment, whereas levels of Bax and catalase were decreased. The endoplasmic reticulum (ER) stress marker GRP78/BiP, IRE1α, and p-PERK proteins were significantly reduced by melatonin treatment. Treatment with the competitive melatonin receptor antagonist luzindole blocked melatonin-induced decreases in LC3 II levels. These results demonstrate that melatonin suppresses clinostat-induced autophagy through increasing the phosphorylation of the ERK/Akt/mTOR proteins. Consequently, melatonin appears to be a potential therapeutic agent for regulating microgravity-related bone loss or osteoporosis. PMID:27070587

  1. Human ubiquitin-activating enzyme, E1. Indication of potential nuclear and cytoplasmic subpopulations using epitope-tagged cDNA constructs.

    PubMed

    Handley-Gearhart, P M; Stephen, A G; Trausch-Azar, J S; Ciechanover, A; Schwartz, A L

    1994-12-30

    The ubiquitin-activating enzyme E1 catalyzes the first step in the ubiquitin conjugation pathway. Previously, we have cloned and sequenced the cDNA for human E1. Expression of the E1 cDNA in the ts20 cell line, which harbors a thermolabile E1, abrogated the phenotypic defects associated with this line. However, little is known of the cell biology of the E1 protein or the nature of the E1 doublet. Thus, we constructed epitope-tagged E1 cDNAs in which the HA monoclonal antibody epitope tag sequence (from influenza hemagglutinin and recognized by the 12CA5 monoclonal antibody) was fused to the amino terminus of E1. Because the amino-terminal amino acid sequence of E1 is unknown, three constructs were made in which the HA tag was placed at each of the first three ATGs in the open reading frame (HA-1E1, HA-2E1, and HA-3E1). Western analysis of HeLa cells transfected with the constructs revealed that HA-1E1 closely comigrated with the upper band of the E1 doublet, and HA-2E1 comigrated with the lower band of the E1 doublet; HA-3E1 appeared smaller than either of the E1 bands. Metabolic labeling with 32P and immunoprecipitation with anti-HA antibody revealed that only the HA-1E1 protein product is phosphorylated; polyclonal anti-E1 antibody showed that only the upper band of the endogenous E1 doublet is phosphorylated. Each of the constructs was able to rescue the mutant phenotype of the ts20 cell line. Immunofluorescence studies showed that HA-2E1 and HA-3E1 were distributed in the cytoplasm with both negative and positive nuclei. This pattern of distribution has also been observed when immunostaining with a monoclonal antibody to E1 (1C5). However, the staining pattern associated with a polyclonal anti-E1 antibody (JJJ) is characterized by positive staining cytoplasm and nuclei in all cells. The HA-1E1 construct exhibited apparently exclusive nuclear distribution in HeLa cells. The difference between the staining patterns of the polyclonal and monoclonal anti-E1 antibodies can be explained by the existence of two subpopulations of E1: one cytoplasmic and partially nuclear, and one that is nuclear. Deletion of a small region at the amino terminus of the HA-1E1, including the basic sequence KKRR, transformed its immunostaining pattern to that observed with HA-2E1. PMID:7528747

  2. Localization of the adenovirus E1Aa protein, a positive-acting transcriptional factor, in infected cells infected cells.

    PubMed Central

    Feldman, L T; Nevins, J R

    1983-01-01

    The function of the adenovirus E1Aa protein (the product of the 13S E1A mRNA) during a productive viral infection is to activate transcription of the six early viral transcription units. To study the mechanism of action of this protein, a peptide which was 13 amino acids long and had a sequence unique to the protein product of the adenovirus 13S E1A mRNA (pE1Aa) was coupled to keyhole limpet hemocyanin and used to raise an antibody in rabbits. The resulting antiserum was specific to this protein and did not react with the protein product of the 12S E1A mRNA, which shares considerable sequence with the E1Aa protein. This antiserum was used to probe for the E1Aa protein in situ by indirect immunofluorescence and in extracts of infected HeLa cells. We found that the protein was associated with large cellular structures both in the nucleus and in the cytoplasm. The nuclear form of the protein was analyzed further and was found to purify with the nuclear matrix. Images PMID:6346057

  3. The Transmembrane Domain of Hepatitis C Virus Glycoprotein E1 Is a Signal for Static Retention in the Endoplasmic Reticulum

    PubMed Central

    Cocquerel, Laurence; Duvet, Sandrine; Meunier, Jean-Christophe; Pillez, André; Cacan, René; Wychowski, Czeslaw; Dubuisson, Jean

    1999-01-01

    Hepatitis C virus (HCV) glycoproteins E1 and E2 assemble to form a noncovalent heterodimer which, in the cell, accumulates in the endoplasmic reticulum (ER). Contrary to what is observed for proteins with a KDEL or a KKXX ER-targeting signal, the ER localization of the HCV glycoprotein complex is due to a static retention in this compartment rather than to its retrieval from the cis-Golgi region. A static retention in the ER is also observed when E2 is expressed in the absence of E1 or for a chimeric protein containing the ectodomain of CD4 in fusion with the transmembrane domain (TMD) of E2. Although they do not exclude the presence of an intracellular localization signal in E1, these data do suggest that the TMD of E2 is an ER retention signal for HCV glycoprotein complex. In this study chimeric proteins containing the ectodomain of CD4 or CD8 fused to the C-terminal hydrophobic sequence of E1 were shown to be localized in the ER, indicating that the TMD of E1 is also a signal for ER localization. In addition, these chimeric proteins were not processed by Golgi enzymes, indicating that the TMD of E1 is responsible for true retention in the ER, without recycling through the Golgi apparatus. Together, these data suggest that at least two signals (TMDs of E1 and E2) are involved in ER retention of the HCV glycoprotein complex. PMID:10074109

  4. Conserved tyrosine 182 residue in hyperthermophilic esterase EstE1 plays a critical role in stabilizing the active site.

    PubMed

    Truongvan, Ngoc; Chung, Hye-Shin; Jang, Sei-Heon; Lee, ChangWoo

    2016-03-01

    An aromatic amino acid, Tyr or Trp, located in the esterase active site wall, is highly conserved, with hyperthermophilic esterases showing preference for Tyr and lower temperature esterases showing preference for Trp. In this study, we investigated the role of Tyr(182) in the active site wall of hyperthermophilic esterase EstE1. Mutation of Tyr to Phe or Ala had a moderate effect on EstE1 thermal stability. However, a small-to-large mutation such as Tyr to His or Trp had a devastating effect on thermal stability. All mutant EstE1 enzymes showed reduced catalytic rates and enhanced substrate affinities as compared with wild-type EstE1. Hydrogen bond formation involving Tyr(182) was unimportant for maintaining EstE1 thermal stability, as the EstE1 structure is already adapted to high temperatures via increased intramolecular interactions. However, removal of hydrogen bond from Tyr(182) significantly decreased EstE1 catalytic activity, suggesting its role in stabilization of the active site. These results suggest that Tyr is preferred over a similarly sized Phe residue or bulky His or Trp residue in the active site walls of hyperthermophilic esterases for stabilizing the active site and regulating catalytic activity at high temperatures. PMID:26838013

  5. The regulation of cytochrome P450 2E1 during LPS-induced inflammation in the rat

    SciTech Connect

    Abdulla, Dalya; Goralski, Kerry B.; Renton, Kenneth W. . E-mail: Ken.Renton@dal.ca

    2006-10-01

    It is well known that inflammatory and infectious conditions differentially regulate cytochrome P450 (P450)-mediated drug metabolism in the liver. We have previously outlined a potential pathway for the downregulation in hepatic cytochrome P450 following LPS-mediated inflammation in the CNS (Abdulla, D., Goralski, K.B., Garcia Del Busto Cano, E., Renton, K.W., 2005. The signal transduction pathways involved in hepatic cytochrome P450 regulation in the rat during an LPS-induced model of CNS inflammation. Drug Metab. Dispos). The purpose of this study was to outline the effects of LPS-induced peripheral and central nervous system inflammation on hepatic cytochrome P450 2E1 (CYP2E1) in vivo, an enzyme that plays an important role in various physiological and pathological states. We report an increase in hepatic mRNA expression of CYP2E1 that occurred as early as 2-3 h following either the intraperitoneal (i.p.) injection of 5 mg/kg LPS or i.c.v. administration of 25 {mu}g of LPS. This increase in CYP2E1 mRNA expression was sustained for 24 h. In sharp contrast to the increase in hepatic CYP2E1 mRNA, we observed a significant reduction in the catalytic activity of this enzyme 24 h following either the i.c.v. or i.p. administration of LPS. Cycloheximide or actinomycin-D did not change the LPS-mediated downregulation in hepatic CYP2E1 catalytic activity. Our results support the idea that LPS acts at two different levels to regulate hepatic CYP2E1: a transcriptional level to increase CYP2E1 mRNA expression and a post-transcriptional level to regulate CYP2E1 protein and activity.

  6. Adenovirus E1A243 disrupts the ATF/CREB-YY1 complex at the mouse c-fos promoter.

    PubMed Central

    Zhou, Q; Engel, D A

    1995-01-01

    The adenovirus E1A243 protein can activate transcription of the mouse c-fos gene in a manner that depends on treatment of cells with inducers or analogs of cyclic AMP (cAMP). Activation requires conserved region 1 and the N-terminal domain of E1A243 and is mediated by a 22-bp E1A response element containing a cAMP response element (CRE) at -67 and a binding site for transcription factor YY1 at -54. In the absence of E1A243, YY1 represses CRE-dependent transcription of c-fos by physically interacting with ATF/CREB proteins bound to the -67 CRE. Here we present evidence that expression of E1A243 leads to relief of YY1-mediated repression by a disruption of the ATF/CREB-YY1 complex. Addition of E1A243 to in vitro binding assays prevented binding of ATF-2 to glutathione S-transferase-YY1. Similarly, expression of E1A243 in HeLa cells prevented the association of a YY1-VP16 fusion protein with endogenous ATF/CREB proteins bound to the -67 CRE of a transfected c-fosCAT reporter plasmid. In each case, the N-terminal domain of E1A243, which mediates a direct interaction with YY1, was responsible for disruption of the ATF/CREB-YY1 complex. On the basis of these and previously published results, we present a model for the synergistic transcriptional activation of the c-fos gene by E1A243 and cAMP. PMID:7494244

  7. Chronic administration of caderofloxacin, a new fluoroquinolone, increases hepatic CYP2E1 expression and activity in rats

    PubMed Central

    Liu, Li; Miao, Ming-xing; Zhong, Ze-yu; Xu, Ping; Chen, Yang; Liu, Xiao-dong

    2016-01-01

    Aim: Caderofloxacin is a new fluoroquinolone that is under phase III clinical trials in China. Here we examined the effects of caderofloxacin on rat hepatic cytochrome P450 (CYP450) isoforms as well as the potential of caderofloxacin interacting with co-administered drugs. Methods: Male rats were treated with caderofloxacin (9 mg/kg, ig) once or twice daily for 14 consecutive days. The effects of caderofloxacin on CYP3A, 2D6, 2C19, 1A2, 2E1 and 2C9 were evaluated using a “cocktail” of 6 probes (midazolam, dextromethorphan, omeprazole, theophylline, chlorzoxazone and diclofenac) injected on d 0 (prior to caderofloxacin exposure) and d 15 (after caderofloxacin exposure). Hepatic microsomes from the caderofloxacin-treated rats were used to assess CYP2E1 activity and chlorzoxazone metabolism. The expression of CYP2E1 mRNA and protein in hepatic microsomes was analyzed with RT-PCR and Western blotting, respectively. Results: Fourteen-day administration of caderofloxacin significantly increased the activity of hepatic CYP2E1, leading to enhanced metabolism of chlorzoxazone. In vitro microsomal study confirmed that CYP2E1 was a major metabolic enzyme involved in chlorzoxazone metabolism, and the 14-d administration of caderofloxacin significantly increased the activity of CYP2E1 in hepatic microsomes, resulting in increased formation of 6-hydroxychlorzoxazone. Furthermore, the 14-d administration of caderofloxacin significantly increased the expression of CYP2E1 mRNA and protein in liver microsomes, which was consistent with the pharmacokinetic results. Conclusion: Fourteen-day administration of caderofloxacin can induce the expression and activity of hepatic CYP2E1 in rats. When caderofloxacin is administered, a potential drug-drug interaction mediated by CYP2E1 induction should be considered. PMID:26838075

  8. Dielectric strength of parylene HT

    SciTech Connect

    Diaham, S. Bechara, M.; Locatelli, M.-L.; Khazaka, R.; Tenailleau, C.

    2014-02-07

    The dielectric strength of parylene HT (PA-HT) films was studied at room temperature in a wide thickness range from 500 nm to 50 μm and was correlated with nano- and microstructure analyses. X-ray diffraction and polarized optical microscopy have revealed an enhancement of crystallization and spherulites development, respectively, with increasing the material thickness (d). Moreover, a critical thickness d{sub C} (between 5 and 10 μm) is identified corresponding to the beginning of spherulite developments in the films. Two distinct behaviors of the dielectric strength (F{sub B}) appear in the thickness range. For d ≥ d{sub C}, PA-HT films exhibit a decrease in the breakdown field following a negative slope (F{sub B} ∼ d{sup −0.4}), while for d < d{sub C}, it increases with increasing the thickness (F{sub B} ∼ d{sup 0.3}). An optimal thickness d{sub optim} ∼ 5 μm corresponding to a maximum dielectric strength (F{sub B} ∼ 10 MV/cm) is obtained. A model of spherulite development in PA-HT films with increasing the thickness is proposed. The decrease in F{sub B} above d{sub C} is explained by the spherulites development, whereas its increase below d{sub C} is induced by the crystallites growth. An annealing of the material shows both an enhancement of F{sub B} and an increase of the crystallites and spherulites dimensions, whatever the thickness. The breakdown field becomes thickness-independent below d{sub C} showing a strong influence of the nano-scale structural parameters. On the contrary, both nano- and micro-scale structural parameters appear as influent on F{sub B} for d ≥ d{sub C}.

  9. High strength, tough alloy steel

    DOEpatents

    Thomas, Gareth; Rao, Bangaru V. N.

    1979-01-01

    A high strength, tough alloy steel is formed by heating the steel to a temperature in the austenite range (1000.degree.-1100.degree. C.) to form a homogeneous austenite phase and then cooling the steel to form a microstructure of uniformly dispersed dislocated martensite separated by continuous thin boundary films of stabilized retained austenite. The steel includes 0.2-0.35 weight % carbon, at least 1% and preferably 3-4.5% chromium, and at least one other substitutional alloying element, preferably manganese or nickel. The austenite film is stable to subsequent heat treatment as by tempering (below 300.degree. C.) and reforms to a stable film after austenite grain refinement.

  10. 14 CFR 31.27 - Strength.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ..., joints, and load attachment points and members. (c) An ultimate free-fall drop test must be made of the... STANDARDS: MANNED FREE BALLOONS Strength Requirements § 31.27 Strength. (a) The structure must be able...

  11. 14 CFR 31.27 - Strength.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., joints, and load attachment points and members. (c) An ultimate free-fall drop test must be made of the... STANDARDS: MANNED FREE BALLOONS Strength Requirements § 31.27 Strength. (a) The structure must be able...

  12. 14 CFR 31.27 - Strength.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ..., joints, and load attachment points and members. (c) An ultimate free-fall drop test must be made of the... STANDARDS: MANNED FREE BALLOONS Strength Requirements § 31.27 Strength. (a) The structure must be able...

  13. 26 CFR 301.6226(e)-1 - Jurisdictional requirement for bringing an action in District Court or United States Court of...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... actions beginning prior to October 4, 2001, see § 301.6226(e)-1T contained in 26 CFR part 1, revised April... action in District Court or United States Court of Federal Claims. 301.6226(e)-1 Section 301.6226(e)-1... ADMINISTRATION PROCEDURE AND ADMINISTRATION Assessment In General § 301.6226(e)-1 Jurisdictional requirement...

  14. Bonded joint strength - Static versus fatigue

    NASA Technical Reports Server (NTRS)

    Johnson, W. S.; Mall, S.

    1984-01-01

    Adhesives are commonly characterized only by their static strength even though they are used in structural joints that are subjected to fatigue loads. This paper reviews the relationship between static and fatigue strength for four different specimen types: single-lap-shear, edge-delamination, double cantilever beam, and cracked-lap-shear. It was found that the ratio of static strength to fatigue strength varied from 2.3 to 4.7, depending on the adhesive and specimen configuration.

  15. Strength distribution in commercial silicon carbide materials

    NASA Technical Reports Server (NTRS)

    Dutta, Sunil

    1988-01-01

    Four-point flexural strength testing has been conducted in order to establish the baseline strength and reliability of four different commercial SiC types, in conjunction with reliable Weibull modulus values. Average strength of the samples ranged from 380 to 482 MPa at room temperature and 307 to 470 MPa at 1370 C. The strength scatter reflects the effect of flaw variability, which must be minimized to improve reliability in sintered SiC.

  16. Strength of Shocked Aluminum Oxynitride

    NASA Astrophysics Data System (ADS)

    Zhu, J.; Feng, R.; Dandekar, D. P.

    2009-06-01

    Aluminum oxynitride (AlON) is a polycrystalline and transparent ceramic. An accurate characterization of its shock response is critically important for its applications as transparent armor. Shock wave profiles measured in a series of plate impact experiments on AlON [Thornhill, et al., SCCM-2005, 143-146 (2006)] have been reanalyzed using finite element wave propagation simulations and considering an effective strength behavior that is pressure- and time-dependent. The results show a stiffer shock response than that calculated previously using the jump conditions. The material has a Hugoniot elastic limit of 10.37 GPa and sustains a maximum shear stress of 4.38 GPa for shock compressions up to a shock stress of 96 GPa. The mean stress response determined from the simulations displays no sign of phase transformation and corresponds to a linear shock speed-particle velocity relation with a slope of 0.857. These results have been successfully summarized into an AlON material model consisting of compression-dependent nonlinear elasticity, pressure-dependent equilibrium strength, and over-stress relaxation. The wave profiles simulated with the model show very good agreement with the experimental measurements.

  17. Frontiers in Strength-Based Treatment

    ERIC Educational Resources Information Center

    Laursen, Erik K.

    2003-01-01

    "Reclaiming Children and Youth's" guest editor introduces this special issue on promising practices employing strength-based approaches to education and treatment. If we are serious about the commitment to "leave no child behind," then we must be able to cultivate strengths in all children and families. This article surveys the strengths

  18. Strengths and Satisfaction across the Adult Lifespan

    ERIC Educational Resources Information Center

    Isaacowitz, Derek M.; Vaillant, George E.; Seligman, Martin E. P.

    2003-01-01

    Positive psychology has recently developed a classification of human strengths (Peterson & Seligman, in press). We aimed to evaluate these strengths by investigating the strengths and life satisfaction in three adult samples recruited from the community (young adult, middle-aged, and older adult), as well as in the surviving men of the Grant study…

  19. 14 CFR 31.27 - Strength.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Strength. 31.27 Section 31.27 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: MANNED FREE BALLOONS Strength Requirements § 31.27 Strength. (a) The structure must be able...

  20. 14 CFR 31.27 - Strength.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Strength. 31.27 Section 31.27 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION, DEPARTMENT OF TRANSPORTATION AIRCRAFT AIRWORTHINESS STANDARDS: MANNED FREE BALLOONS Strength Requirements § 31.27 Strength. (a) The structure must be able...

  1. Improving the toughness of ultrahigh strength steel

    SciTech Connect

    Soto, Koji

    2002-08-15

    The ideal structural steel combines high strength with high fracture toughness. This dissertation discusses the toughening mechanism of the Fe/Co/Ni/Cr/Mo/C steel, AerMet 100, which has the highest toughness/strength combination among all commercial ultrahigh strength steels. The possibility of improving the toughness of this steel was examined by considering several relevant factors.

  2. 7 CFR 29.3061 - Strength (tensile).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Strength (tensile). 29.3061 Section 29.3061... Type 93) § 29.3061 Strength (tensile). The stress a tobacco leaf can bear without tearing. Tensile strength is not an important element of quality in Burley tobacco....

  3. ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation.

    PubMed

    Freudenau, Inga; Lutter, Petra; Baier, Ruth; Schleef, Martin; Bednarz, Hanna; Lara, Alvaro R; Niehaus, Karsten

    2015-01-01

    Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the past years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3) and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ) by using ordinary differential equations and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII) as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN), and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI- and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ± 0.7 and 34 ± 7 RNAI molecules per cell and 0.44 ± 0.1 and 3 ± 0.9 RNAII molecules per cell. The determined PCNs averaged between 46 ± 26 and 48 ± 30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ± 203 and 1086 ± 298 RNAI molecules per cell and 22 ± 2 and 75 ± 10 RNAII molecules per cell with an averaged PCN of 1514 ± 1301 and 5806 ± 4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the plasmid DNA production. PMID:26389114

  4. ColE1-Plasmid Production in Escherichia coli: Mathematical Simulation and Experimental Validation

    PubMed Central

    Freudenau, Inga; Lutter, Petra; Baier, Ruth; Schleef, Martin; Bednarz, Hanna; Lara, Alvaro R.; Niehaus, Karsten

    2015-01-01

    Plasmids have become very important as pharmaceutical gene vectors in the fields of gene therapy and genetic vaccination in the past years. In this study, we present a dynamic model to simulate the ColE1-like plasmid replication control, once for a DH5α-strain carrying a low copy plasmid (DH5α-pSUP 201-3) and once for a DH5α-strain carrying a high copy plasmid (DH5α-pCMV-lacZ) by using ordinary differential equations and the MATLAB software. The model includes the plasmid replication control by two regulatory RNA molecules (RNAI and RNAII) as well as the replication control by uncharged tRNA molecules. To validate the model, experimental data like RNAI- and RNAII concentration, plasmid copy number (PCN), and growth rate for three different time points in the exponential phase were determined. Depending on the sampled time point, the measured RNAI- and RNAII concentrations for DH5α-pSUP 201-3 reside between 6 ± 0.7 and 34 ± 7 RNAI molecules per cell and 0.44 ± 0.1 and 3 ± 0.9 RNAII molecules per cell. The determined PCNs averaged between 46 ± 26 and 48 ± 30 plasmids per cell. The experimentally determined data for DH5α-pCMV-lacZ reside between 345 ± 203 and 1086 ± 298 RNAI molecules per cell and 22 ± 2 and 75 ± 10 RNAII molecules per cell with an averaged PCN of 1514 ± 1301 and 5806 ± 4828 depending on the measured time point. As the model was shown to be consistent with the experimentally determined data, measured at three different time points within the growth of the same strain, we performed predictive simulations concerning the effect of uncharged tRNA molecules on the ColE1-like plasmid replication control. The hypothesis is that these tRNA molecules would have an enhancing effect on the plasmid production. The in silico analysis predicts that uncharged tRNA molecules would indeed increase the plasmid DNA production. PMID:26389114

  5. Artificial Recruitment of UAF1-USP Complexes by a PHLPP1-E1 Chimeric Helicase Enhances Human Papillomavirus DNA Replication

    PubMed Central

    Gagnon, David; Lehoux, Michaël

    2015-01-01

    ABSTRACT The E1 helicase from anogenital human papillomavirus (HPV) types interacts with the cellular WD repeat-containing protein UAF1 in complex with the deubiquitinating enzyme USP1, USP12, or USP46. This interaction stimulates viral DNA replication and is required for maintenance of the viral episome in keratinocytes. E1 associates with UAF1 through a short UAF1-binding site (UBS) located within the N-terminal 40 residues of the protein. Here, we investigated if the E1 UBS could be replaced by the analogous domain from an unrelated protein, the pleckstrin homology domain and leucine-rich repeat protein phosphatase 1 (PHLPP1). We found that PHLPP1 and E1 interact with UAF1 in a mutually exclusive manner and mapped the minimal PHLPP1 UBS (PUBS) to a 100-amino-acid region sufficient for assembly into UAF1-USP complexes. Similarly to the E1 UBS, overexpression of PUBS in trans inhibited HPV DNA replication, albeit less efficiently. Characterization of a PHLPP1-E1 chimeric helicase revealed that PUBS could partially substitute for the E1 UBS in enhancing viral DNA replication and that the stimulatory effect of PUBS likely involves recruitment of UAF1-USP complexes, as it was abolished by mutations that weaken UAF1-binding and by overexpression of catalytically inactive USPs. Although functionally similar to the E1 UBS, PUBS is larger in size and requires both the WD repeat region and C-terminal ubiquitin-like domain of UAF1 for interaction, in contrast to E1, which does not contact the latter. Overall, this comparison of two heterologous UBSs indicates that these domains function as transferable protein interaction modules and provide further evidence that the association of E1 with UAF1-containing deubiquitinating complexes stimulates HPV DNA replication. IMPORTANCE The E1 protein from anogenital HPV types interacts with the UAF1-associated deubiquitinating enzymes USP1, USP12, and USP46 to stimulate replication of the viral genome. Little is known about the molecular nature of the E1-UAF1 interaction and, more generally, how UAF1-USP complexes recognize their substrate proteins. To address this question, we characterized the UAF1-binding site (UBS) of PHLPP1, a protein unrelated to E1. Using a PHLPP1-E1 chimeric helicase, we show that the PHLPP1 UBS (PUBS) can partially substitute for the E1 UBS in stimulating HPV DNA replication. This stimulation required conserved sequences in PUBS that meditate its interaction with UAF1, including a motif common to the E1 UBS. These results indicate that UAF1-binding sequences function as transferable protein interaction modules and provide further evidence that UAF1-USP complexes stimulate HPV DNA replication. PMID:25833051

  6. CYP2E1 overexpression inhibits microsomal Ca2+-ATPase activity in HepG2 cells.

    PubMed

    Caro, Andres A; Evans, Kerry L; Cederbaum, Arthur I

    2009-01-31

    Cytochrome P450 2E1 (CYP2E1) is a microsomal enzyme that generates reactive oxygen species during its catalytic cycle. We previously found an important role for calcium in CYP2E1-potentiated injury in HepG2 cells. The possibility that CYP2E1 may oxidatively damage and inactivate the microsomal Ca2+-ATPase in intact liver cells was evaluated, in order to explain why calcium is elevated during CYP2E1 toxicity. Microsomes were isolated by differential centrifugation from two liver cell line: E47 cells (HepG2 cells transfected with the pCI neo expression vector containing the human CYP2E1 cDNA, which overexpress active microsomal CYP2E1), and control C34 cells (HepG2 cells transfected with the pCI neo expression vector alone, which do not express significantly any cytochrome P450). The Ca2+-dependent ATPase activity was determined by measuring the accumulation of inorganic phosphate from ATP hydrolysis. CYP2E1 overexpression produced a 45% decrease in Ca2+-dependent ATPase activity (8.6 nmol Pi/min/mg protein in C34 microsomes versus 4.7 nmol Pi/min/mg protein in microsomes). Saturation curves with Ca2+ or ATP showed that CYP2E1 overexpression produced a decrease in Vmax but did not affect the Km for either Ca2+ or ATP. The decrease in activity was not associated with a decrease in SERCA protein levels. The ATP-dependent microsomal calcium uptake was evaluated by fluorimetry using fluo-3 as the fluorogenic probe. Calcium uptake rate in E47 microsomes was 28% lower than in C34 microsomes. Treatment of E47 cells with 2mM N-acetylcysteine prevented the decrease in microsomal Ca2+-ATPase found in E47 cells. These results suggest that CYP2E1 overexpression produces a decrease in microsomal Ca2+-ATPase activity in HepG2 cells mediated by reactive oxygen species. This may contribute to elevated cytosolic calcium and to CYP2E1-potentiated injury. PMID:19028543

  7. CYP2E1 epigenetic regulation in chronic, low-level toluene exposure: Relationship with oxidative stress and smoking habit

    SciTech Connect

    Jiménez-Garza, Octavio; Baccarelli, Andrea A.; Byun, Hyang-Min; Márquez-Gamiño, Sergio; Barrón-Vivanco, Briscia Socorro

    2015-08-01

    Background: CYP2E1 is a versatile phase I drug-metabolizing enzyme responsible for the biotransformation of most volatile organic compounds, including toluene. Human toluene exposure increases CYP2E1 mRNA and modifies its activity in leucocytes; however, epigenetic implications of this interaction have not been investigated. Goal: To determine promoter methylation of CYP2E1 and other genes known to be affected by toluene exposure. Methods: We obtained venous blood from 24 tannery workers exposed to toluene (mean levels: 10.86 +/− 7 mg/m{sup 3}) and 24 administrative workers (reference group, mean levels 0.21 +/− 0.02 mg/m{sup 3}) all of them from the city of León, Guanajuato, México. After DNA extraction and bisulfite treatment, we performed PCR-pyrosequencing in order to measure methylation levels at promoter region of 13 genes. Results: In exposed group we found significant correlations between toluene airborne levels and CYP2E1 promoter methylation (r = − .36, p < 0.05), as well as for IL6 promoter methylation levels (r = .44, p < 0.05). Moreover, CYP2E1 promoter methylation levels where higher in toluene-exposed smokers compared to nonsmokers (p = 0.009). We also observed significant correlations for CYP2E1 promoter methylation with GSTP1 and SOD1 promoter methylation levels (r = − .37, p < 0.05 and r = − .34, p < 0.05 respectively). Conclusion: These results highlight the importance of considering CYP2E1 epigenetic modifications, as well as its interactions with other genes, as key factors for unraveling the sub cellular mechanisms of toxicity exerted by oxidative stress, which can initiate disease process in chronic, low-level toluene exposure. People co-exposed to toluene and tobacco smoke are in higher risk due to a possible CYP2E1 repression. - Highlights: • We investigated gene-specific methylation in persons chronically exposed to toluene. • In a previous study, a reduced CYP2E1 activity was observed in these participants. • CYP2E1 promoter methylation correlated with oxidative-stress related gene methylation. • CYP2E1 promoter methylation was higher in exposed smokers compared to nonsmokers. • Epigenetic modifications are critical to detect early toluene-exposure effects.

  8. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 11 Federal Elections 1 2012-01-01 2012-01-01 false Non-Federal elections (2 U.S.C. 441i(e)(1)(B)). 300.62 Section 300.62 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive,...

  9. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Federal elections (2 U.S.C. 441i(e)(1)(A)). 300.61 Section 300.61 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive,...

  10. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 11 Federal Elections 1 2014-01-01 2014-01-01 false Non-Federal elections (2 U.S.C. 441i(e)(1)(B)). 300.62 Section 300.62 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive,...

  11. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 11 Federal Elections 1 2013-01-01 2012-01-01 true Non-Federal elections (2 U.S.C. 441i(e)(1)(B)). 300.62 Section 300.62 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive,...

  12. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 11 Federal Elections 1 2014-01-01 2014-01-01 false Federal elections (2 U.S.C. 441i(e)(1)(A)). 300.61 Section 300.61 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive,...

  13. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 11 Federal Elections 1 2013-01-01 2012-01-01 true Federal elections (2 U.S.C. 441i(e)(1)(A)). 300.61 Section 300.61 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive,...

  14. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 11 Federal Elections 1 2012-01-01 2012-01-01 false Federal elections (2 U.S.C. 441i(e)(1)(A)). 300.61 Section 300.61 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive,...

  15. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 11 Federal Elections 1 2011-01-01 2011-01-01 false Non-Federal elections (2 U.S.C. 441i(e)(1)(B)). 300.62 Section 300.62 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive,...

  16. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 11 Federal Elections 1 2011-01-01 2011-01-01 false Federal elections (2 U.S.C. 441i(e)(1)(A)). 300.61 Section 300.61 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive,...

  17. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 11 Federal Elections 1 2010-01-01 2010-01-01 false Non-Federal elections (2 U.S.C. 441i(e)(1)(B)). 300.62 Section 300.62 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive,...

  18. Cellular localization studies on human estrogen sulfotransferase SULT1E1 in human embryonic kidney 293 cells.

    PubMed

    Kapoor, Ruchita; Nimmagadda, Deepthi; Sheng, Jonathan J

    2007-01-01

    Human cytosolic sulfotransferase SULT1E1 catalyzes the sulfation of endogenous estrogens as well as xenobiotic estrogen-like chemicals. This reaction increases the water solubility of the molecule, which may affect its cellular distribution and biological activity. This could alter estrogen signaling to the estrogen receptor in human estrogen receptor-positive cells. The current work characterized the cellular distribution of SULT1E1 in the human embryonic kidney 293 (HEK293) cell line using green fluorescent protein (GFP) tagging and immunochemistry methods. The GFP-tagged recombinant SULT1E1 protein was expressed and localized in the cytoplasm of HEK293 cells. By using a commercial anti-SULT1E1 peptide antibody, a 35.7-kDa protein was detected in HEK293 cells via Western blot. The molecular mass of the protein detected suggested that it may be related to native SULT1E1 protein. However, reverse transcription-polymerase chain reaction (RT-PCR) with gene-specific primers could not confirm the presence of the SULT1E1 transcript in the total RNA sample of HEK293 cells. The discrepancy between protein and transcript data could be due to the instability of SULT1E1 mRNA or the specificity of the anti-SULT1E1 antibody used. In the present work, RT-PCR analysis with gene-specific primers also identified a transcript fragment of human estrogen-related receptor gamma. Future studies on the functional relationship between estrogen-related receptors and sulfotransferases are expected to provide additional insights into the physiological and toxicological roles of human estrogen sulfotransferases. PMID:17035602

  19. Modulation of p53-mediated transcriptional repression and apoptosis by the adenovirus E1B 19K protein.

    PubMed Central

    Sabbatini, P; Chiou, S K; Rao, L; White, E

    1995-01-01

    BRK cell lines that stably express adenovirus E1A and a murine temperature-sensitive p53 undergo apoptosis when p53 assumes the wild-type conformation. Expression of the E1B 19,000-molecular-weight (19K) protein rescues cells from this p53-mediated apoptosis and diverts cells to a growth-arrested state. As p53 likely functions as a tumor suppressor by regulating transcription, the ability of the E1B 19K protein to regulate p53-mediated transactivation and transcriptional repression was investigated. In promoter-reporter assays the E1B 19K did not block p53-mediated transactivation but did alleviate p53-mediated transcriptional repression. E1B 19K expression permitted efficient transcriptional activation of the p21/WAF-1/cip-1 mRNA by p53, consistent with maintenance of the growth arrest function of p53. The E1B 19K protein is thereby unique among DNA virus-transforming proteins that target p53 for inactivation in that it selectively modulates the transcriptional properties of p53. The E1B 19K protein also rescued cells from apoptosis induced by inhibitors of transcription and protein synthesis. This suggests that cell death may result from the inhibition of expression of survival factors which function to maintain cell viability. p53 may induce apoptosis through generalized transcriptional repression. In turn, the E1B 19K protein may prevent p53-mediated apoptosis by alleviating p53-mediated transcriptional repression. PMID:7823921

  20. Human CYP2E1-dependent mutagenicity of mono- and dichlorobiphenyls in Chinese hamster (V79)-derived cells.

    PubMed

    Zhang, Chiteng; Lai, Yanmei; Jin, Guifang; Glatt, Hansruedi; Wei, Qinzhi; Liu, Yungang

    2016-02-01

    Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants with confirmed carcinogenicity to humans. Metabolic activation of lower chlorinated PCBs to genotoxic metabolites may involve hydroxylation and further oxidation, and some hydroxylated metabolites may be sulfo-conjugated. However, the genotoxicity of individual PCB compounds is largely unknown. In this study, 15 mono- and dichlorobiphenyls were investigated for genotoxicity using the micronucleus and Hprt mutagenicity assays in a Chinese hamster V79-derived cell line expressing both human cytochrome P450 (CYP) 2E1 and human sulfotransferase (SULT) 1A1 (V79-hCYP2E1-hSULT1A1). All tested compounds were inactive in both assays in V79 control cells. However, eight dichlorobiphenyls strongly induced micronuclei and other congeners were weakly positive for this endpoint in V79-hCYP2E1-hSULT1A1 cells. The effects of each PCB in V79-hCYP2E1-hSULT1A1 cells were abolished or reduced in the presence of a CYP2E1 inhibitor (1-aminobenzotriazole), or enhanced by pretreatment of the cells with (CYP2E1-inducing) ethanol, while the genotoxicity was not significantly affected by a SULT1 inhibitor (pentachlorophenol). As representative dichlorobiphenyls, PCB 5, 10, 8 and 11 (2,3-, 2,5-, 2,4'- and 3,3'-dichlorobiphenyl, respectively) strongly induced Hprt gene mutations in V79-hCYP2E1-hSULT1A1 cells in a concentration-dependent manner. This is the first indication that human CYP2E1 is capable of converting a series of dichlorobiphenyls to strong mutagens. PMID:26547025

  1. Cytochrome P4502E1, oxidative stress, JNK, and autophagy in acute alcohol-induced fatty liver.

    PubMed

    Yang, Lili; Wu, Defeng; Wang, Xiaodong; Cederbaum, Arthur I

    2012-09-01

    Binge alcohol drinking induces hepatic steatosis. Recent studies showed that chronic ethanol-induced fatty liver was, at least in part, CYP2E1 dependent. The mechanism of acute alcohol-induced steatosis and whether CYP2E1 plays any role are still unclear. Increasing oxidative stress by alcohol can activate the JNK MAP kinase signaling pathway, suggesting that JNK might be a target for prevention of alcohol-induced steatosis. We used CYP2E1 knockout (KO) mice, a JNK inhibitor, and JNK1 or JNK2 knockout mice to test the role of CYP2E1, JNK, and the individual role of JNK1 and JNK2 in acute alcohol-induced steatosis. In wild-type (WT) mice, acute alcohol activates CYP2E1 and increases oxidative stress, which reciprocally increases activation of the JNK signaling pathway. Acute alcohol-induced fatty liver and oxidative stress were blunted in CYP2E1 KO mice and by the JNK inhibitor in WT mice. The antioxidant N-acetylcysteine decreased the acute alcohol-induced oxidative stress, the activation of JNK, and the steatosis but not the activation of CYP2E1. Acute alcohol decreased autophagy and increased expression of SREBP, effects blocked by the JNK inhibitor. Acute alcohol-induced fatty liver was the same in JNK1 and JNK2 KO mice as in WT mice; thus either JNK1 or JNK2 per se is sufficient for induction of steatosis by acute alcohol. The results show that acute alcohol elevation of CYP2E1, oxidative stress, and activation of JNK interact to lower autophagy and increase lipogenic SREBP resulting in fatty liver. PMID:22749809

  2. Regulation of cell cycle of hepatocellular carcinoma by NF90 through modulation of cyclin E1 mRNA stability.

    PubMed

    Jiang, W; Huang, H; Ding, L; Zhu, P; Saiyin, H; Ji, G; Zuo, J; Han, D; Pan, Y; Ding, D; Ma, X; Zhang, Y; Wu, J; Yi, Q; Liu, J O; Huang, H; Dang, Y; Yu, L

    2015-08-20

    Activation of cyclin E1, a key regulator of the G1/S cell-cycle transition, has been implicated in many cancers including hepatocellular carcinoma (HCC). Although much is known about the regulation of cyclin E1 expression and stability, its post-transcriptional regulation mechanism remains incompletely understood. Here, we report that nuclear factor 90 (NF90), a double-stranded RNA (dsRNA) binding protein, regulates cyclin E1 in HCC. We demonstrate that NF90 is upregulated in HCC specimens and that suppression of NF90 decreases HCC cell growth and delays G1/S transition. We identified cyclin E1 as a new target of NF90 and found a significant correlation between NF90 and cyclin E1 expression in HCC. The mRNA and protein levels of cyclin E1 were downregulated upon NF90 knockdown. Suppression of NF90 caused a decrease in the half-life of cyclin E1 mRNA, which was rescued by ectopic expression of NF90. Furthermore, NF90 bound to the 3' untranslated regions (3'UTRs) of cyclin E1 mRNA in vitro and in vivo. Knockdown of NF90 also inhibited tumor growth of HCC cell lines in mouse xenograft model. Moreover, we showed that inhibition of NF90 sensitized HCC cells to the cyclin-dependent kinase 2 (CDK2) inhibitor, roscovitine. Taken together, downregulation of NF90 in HCC cell lines can delay cell-cycle progression, inhibit cell proliferation, and reduce tumorigenic capacity in vivo. These results suggest that NF90 has an important role in HCC pathogenesis and that it can serve as a novel therapeutic target for HCC. PMID:25399696

  3. Complete Genome Sequence of Methanosphaerula palustris E1-9CT, a Hydrogenotrophic Methanogen Isolated from a Minerotrophic Fen Peatland.

    PubMed

    Cadillo-Quiroz, Hinsby; Browne, Patrick; Kyrpides, Nikos; Woyke, Tanja; Goodwin, Lynne; Detter, Chris; Yavitt, Joseph B; Zinder, Stephen H

    2015-01-01

    Here, we report the complete genome sequence (2.92Mb) of Methanosphaerula palustris E1-9C(T), a methanogen isolated from a minerotrophic fen. This is the first genome report of the Methanosphaerula genus, within the Methanoregulaceae family, in the Methanomicrobiales order. E1-9C(T) relatives are found in a wide range of ecological and geographical settings. PMID:26543115

  4. Densification, microstructure and strength evolution in sintering

    NASA Astrophysics Data System (ADS)

    Xu, Xiaoping

    2000-10-01

    Powder metallurgy has the ability to fabricate high quality, complex components to close tolerances in an economical manner. In many applications, a high sintered density is desirable for an improved performance. However, sintering to a high density demands a large shrinkage, often resulting in difficulties with dimensional control. Recent studies indicate the occurrence of a sufficient densification requires a low in situ strength at high sintering temperatures. On the other hand, the low in situ strength often leads to component's distortion in response to the external forces, such as gravity. Unfortunately, lack of knowledge on strength evolution in sintering has been a major challenge to achieve an optimized combination of densification and shape retention. Therefore, the present study investigates strength evolution in sintering and the effects of processing factors. Experiments are performed on prealloyed bronze and elemental mixture of Fe-2Ni powders. For the bronze, a loose casting method is used to fabricate transverse rupture bars, while bars are injection molded for the Fe-2Ni. The in situ transverse rupture strength is measured using the Penn State Flaming Tensile Tester. Experimental results indicate a dependence of densification and strength on sintering temperature. High temperatures enhance densification and interparticle bonding, resulting in strong sintered structures. However, a low in situ strength at high test temperatures indicates the dominance of thermal softening. A strength model combining sintering theories and microstructural parameters is developed to predict both the in situ strength and the post-sintering strength. The model demonstrates the strength of the sintered materials depends on the inherent material strength, the square of neck size ratio, sintered density, and thermal softening. The model is verified by comparison of model predictions with experimental data of the bronze and Fe-2Ni. Compared to prior strength models, this model has certain advantages. It is a predictive model for both the in situ strength and post-sintering strength, and can be extended to other systems.

  5. Nondestructive Determination of Bond Strength

    NASA Technical Reports Server (NTRS)

    2000-01-01

    Although many nondestructive techniques have been applied to detect disbonds in adhesive joints, no absolutely reliable nondestructive method has been developed to detect poor adhesion and evaluate the strength of bonded joints prior to the present work which used nonlinear ultrasonic methods to investigate adhesive bond cure conditions. Previously, a variety of linear and nonlinear ultrasonic methods with water coupling had been used to study aluminum-adhesive-aluminum laminates, prepared under different adhesive curing conditions, for possible bond strength determination. Therefore, in the course of this research effort, a variety of finite-amplitude experimental methods which could possibly differentiate various cure conditions were investigated, including normal and oblique incidence approaches based on nonlinear harmonic generation as well as several non-collinear two-wave interaction approaches. Test samples were mechanically scanned in various ways with respect to the focus of a transmitting transducer operated at several variable excitation frequencies and excitation levels. Even when powerful sample-related resonances were exploited by means of a frequency scanning approach, it was very difficult to isolate the nonlinear characteristics of adhesive bonds. However, a multi-frequency multi-power approach was quite successful and reliable. Ultrasonic tone burst signals at increasing power levels, over a wide frequency range, were transmitted through each bond specimen to determine its excitation dependent nonlinear harmonic resonance behavior. Relative amplitude changes were observed particularly in the higher harmonic spectral data and analyzed using a local displacement and strain analysis in the linear approximation. Two analysis approaches of the excitation-dependent data at specific resonances were found to be quite promising. One of these approaches may represent a very robust algorithm for classifying an adhesive bond as being properly cured or not. Another approach, in addition to differentiation between various cure conditions, may even provide information with respect to the bond strength. Several technical papers were published during the course of this research and a summary is presented in the Ph.D. dissertation of Tobias P. Berndt, a graduate student financially supported by this NASA Grant.

  6. Structure and biochemical analysis of the heparin-induced E1 dimer of the amyloid precursor protein

    PubMed Central

    Dahms, Sven O.; Hoefgen, Sandra; Roeser, Dirk; Schlott, Bernhard; Gührs, Karl-Heinz; Than, Manuel E.

    2010-01-01

    The amyloid precursor protein (APP) is the key player in Alzheimer’s disease pathology, yet APP and its analogues are also essential for neuronal development and cell homeostasis in mammals. We have determined the crystal structure of the entire N-terminal APP-E1 domain consisting of the growth factor like and the copper binding domains at 2.7-Å resolution and show that E1 functions as a rigid functional entity. The two subdomains interact tightly in a pH-dependent manner via an evolutionarily conserved interface area. Two E1 entities dimerize upon their interaction with heparin, requiring 8–12 sugar rings to form the heparin-bridged APP-E1 dimer in an endothermic and pH-dependent process that is characterized by a low micromolar dissociation constant. Limited proteolysis confirms that the heparin-bridged E1 dimers obtained in solution correspond to a dimer contact in our crystal, enabling us to model this heparin-[APP-E1]2 complex. Correspondingly, the APP-based signal transduction, cell–cell- and/or cell–ECM interaction should depend on dimerization induced by heparin, as well as on pH, arguing that APP could fulfill different functions depending on its (sub)cellular localization. PMID:20212142

  7. In Silico Prediction of Human Sulfotransferase 1E1 Activity Guided by Pharmacophores from Molecular Dynamics Simulations.

    PubMed

    Rakers, Christin; Schumacher, Fabian; Meinl, Walter; Glatt, Hansruedi; Kleuser, Burkhard; Wolber, Gerhard

    2016-01-01

    Acting during phase II metabolism, sulfotransferases (SULTs) serve detoxification by transforming a broad spectrum of compounds from pharmaceutical, nutritional, or environmental sources into more easily excretable metabolites. However, SULT activity has also been shown to promote formation of reactive metabolites that may have genotoxic effects. SULT subtype 1E1 (SULT1E1) was identified as a key player in estrogen homeostasis, which is involved in many physiological processes and the pathogenesis of breast and endometrial cancer. The development of an in silico prediction model for SULT1E1 ligands would therefore support the development of metabolically inert drugs and help to assess health risks related to hormonal imbalances. Here, we report on a novel approach to develop a model that enables prediction of substrates and inhibitors of SULT1E1. Molecular dynamics simulations were performed to investigate enzyme flexibility and sample protein conformations. Pharmacophores were developed that served as a cornerstone of the model, and machine learning techniques were applied for prediction refinement. The prediction model was used to screen the DrugBank (a database of experimental and approved drugs): 28% of the predicted hits were reported in literature as ligands of SULT1E1. From the remaining hits, a selection of nine molecules was subjected to biochemical assay validation and experimental results were in accordance with the in silico prediction of SULT1E1 inhibitors and substrates, thus affirming our prediction hypotheses. PMID:26542807

  8. Evidence against a role for serine 129 in determining murine cytochrome P450 Cyp2e-1 protein levels.

    PubMed

    Freeman, J E; Wolf, C R

    1994-11-29

    The cytochrome P450 CYP2E subfamily plays a central role in drug and carcinogen metabolism. The cellular content of this protein is regulated at both the transcriptional and posttranslational levels. CYP2E1 is degraded by both rapid and slow acting proteolytic systems. In the presence of a substrate, CYP2E1 becomes stabilized, and the contribution of the rapid actinig proteolytic pathway to its destruction decreases. It has been suggested that phosphorylation at serine 129 acts as a switch to initiate the fast acting degradative pathway. Phosphorylation at serine 129 has also been suggested to be the point at which hormones, such as insulin, exert actions on the stability of this protein. In order to investigate the role of phosphorylation in determining murine Cyp2e-1 levels, serine 129 was changed by site-directed mutagenesis to amino acids that could not be phosphorylated and the recombinant proteins expressed in COS 7 cells. Replacement of serine 129 with alanine and glycine does not lead to Cyp2e-1 accumulation. In the presence of insulin, although Cyp2e-1 levels increase slightly, specific stabilization of the wild-type protein relative to the two mutant forms is not observed. These observations provide evidence that insulin can act by stabilization of Cyp2e-1 protein but suggest that the phosphorylation of serine 129 is not the molecular basis of stabilization observed. PMID:7947804

  9. Structure of Hepatitis C Virus Envelope Glycoprotein E1 Antigenic Site 314-324 in Complex with Antibody IGH526.

    PubMed

    Kong, Leopold; Kadam, Rameshwar U; Giang, Erick; Ruwona, Tinashe B; Nieusma, Travis; Culhane, Jeffrey C; Stanfield, Robyn L; Dawson, Philip E; Wilson, Ian A; Law, Mansun

    2015-08-14

    Hepatitis C virus (HCV) is a positive-strand RNA virus within the Flaviviridae family. The viral "spike" of HCV is formed by two envelope glycoproteins, E1 and E2, which together mediate viral entry by engaging host receptors and undergoing conformational changes to facilitate membrane fusion. While E2 can be readily produced in the absence of E1, E1 cannot be expressed without E2 and few reagents, including monoclonal antibodies (mAbs), are available for study of this essential HCV glycoprotein. A human mAb to E1, IGH526, was previously reported to cross-neutralize different HCV isolates, and therefore, we sought to further characterize the IGH526 neutralizing epitope to obtain information for vaccine design. We found that mAb IGH526 bound to a discontinuous epitope, but with a major component corresponding to E1 residues 314-324. The crystal structure of IGH526 Fab with this E1 glycopeptide at 1.75Å resolution revealed that the antibody binds to one face of an α-helical peptide. Single mutations on the helix substantially lowered IGH526 binding but did not affect neutralization, indicating either that multiple mutations are required or that additional regions are recognized by the antibody in the context of the membrane-associated envelope oligomer. Molecular dynamics simulations indicate that the free peptide is flexible in solution, suggesting that it requires stabilization for use as a candidate vaccine immunogen. PMID:26135247

  10. A dual inhibition: microRNA-552 suppresses both transcription and translation of cytochrome P450 2E1.

    PubMed

    Miao, Lingling; Yao, Hailan; Li, Chenggang; Pu, Mengfan; Yao, Xuan; Yang, Hui; Qi, Xinming; Ren, Jin; Wang, Yizheng

    2016-04-01

    MicroRNAs (miRNAs) can direct post-transcriptional or transcriptional gene silencing. Here, we report that miR-552 is in the nucleus and cytosol and inhibits human cytochrome P450 (CYP) 2E1 expression at both transcriptional and post-transcriptional levels. MiR-552 via its non-seed sequence forms hybrids with a loop hairpin of the cruciform structure in CYP2E1 promoter region to inhibit SMARCE1 and RNA polymerase II binding to the promoter and CYP2E1 transcription. Expressing SMARCE1 reverses the inhibitory effects of miR-552 on CYP2E1 mRNA expression. MiR-552 with mutations in non-seed region losses its transcriptional, but retains its post-transcriptional repression to CYP2E1. In contrast, mutation in miR-552 seed sequence suppresses its inhibitory effects on CYP2E1 expression at protein, but not at mRNA, levels. Our results suggest that miR-552 is a miRNA with a dual inhibitory ability at transcriptional and post-transcriptional levels leading to an effective inhibition. PMID:26926595

  11. Effect of nafazatrom and indomethacin on pulmonary removal of prostaglandin E1 after endotoxin in rabbits.

    PubMed Central

    Gillis, C. N.; Havill, A. M.; Moalli, R.

    1987-01-01

    1 We compared the effects of endotoxin on pulmonary prostaglandin E1 (PGE1) removal in groups of rabbits pretreated with the cyclo-oxygenase inhibitor, indomethacin, or nafazatrom (Bay g 6575), which has been shown to increase plasma prostacyclin concentrations. 2 In untreated animals, endotoxin transiently decreased pulmonary removal of [3H]-PGE1, caused pulmonary hypertension, systemic hypotension and increased plasma concentrations of PGE2 and 6-keto-PGF1 alpha. 3 Indomethacin pretreatment prevented the transient decrease in pulmonary removal of [3H]-PGE1 in response to endotoxin, prevented the haemodynamic effects and inhibited prostaglandin synthesis. Pretreatment with nafazatrom did not affect the decreased pulmonary removal of [3H]-PGE1, exacerbated the haemodynamic response, reduced survival and potentiated the increase in circulating 6-keto-PGF1 alpha. 4 We conclude that indomethacin acts to prevent the depression of pulmonary [3H]-PGE1 removal by eliminating surface area changes associated with endotoxin-induced pulmonary vasoconstriction. 5 These data suggest that nafazatrom treatment results in exacerbation of the endotoxin-induced systemic hypotension presumably due to its effect on increased plasma prostacyclin during the later phase of endotoxaemia. PMID:3311263

  12. Vibrational studies on (E)-1-((pyridine-2-yl)methylene)semicarbazide using experimental and theoretical method

    NASA Astrophysics Data System (ADS)

    Subashchandrabose, S.; Ramesh Babu, N.; Saleem, H.; Syed Ali Padusha, M.

    2015-08-01

    The (E)-1-((pyridine-2-yl)methylene)semicarbazide (PMSC) was synthesized. The experimental and theoretical study on molecular structure and vibrational spectra were carried out. The FT-IR (400-4000 cm-1), FT-Raman (50-3500 cm-1) and UV-Vis (200-500 nm) spectra of PMSC were recorded. The geometric structure, conformational analysis, vibrational wavenumbers of PMSC in the ground state have been calculated using B3LYP method of 6-311++G(d,p) basis set. The complete vibrational assignments were made on the basis of TED, calculated by SQM method. The Non-linear optical activity was measured by means of first order hyperpolarizability calculation and π-electrons of conjugative bond in the molecule. The intra-molecular charge transfer, mode hyperconjugative interaction and molecular stabilization energies were calculated. The band gap energies between occupied and unoccupied molecular orbitals were analyzed; it proposes lesser band gap with more reactivity. To understand the electronic properties of this molecule the Mulliken charges were also calculated.

  13. Modulation of Osteogenesis in MC3T3-E1 Cells by Different Frequency Electrical Stimulation

    PubMed Central

    Wang, Yu; Cui, Haitao; Wu, Zhenxu; Wu, Naipeng; Wang, Zongliang; Chen, Xuesi; Wei, Yen; Zhang, Peibiao

    2016-01-01

    Electrical stimulation (ES) is therapeutic to many bone diseases, from promoting fracture regeneration to orthopedic intervention. The application of ES offers substantial therapeutic potential, while optimal ES parameters and the underlying mechanisms responsible for the positive clinical impact are poorly understood. In this study, we assembled an ES cell culture and monitoring device. Mc-3T3-E1 cells were subjected to different frequency to investigate the effect of osteogenesis. Cell proliferation, DNA synthesis, the mRNA levels of osteosis-related genes, the activity of alkaline phosphatase (ALP), and intracellular concentration of Ca2+ were thoroughly evaluated. We found that 100 Hz could up-regulate the mRNA levels of collagen I, collagen II and Runx2. On the contrary, ES could down-regulate the mRNA levels of osteopontin (OPN). ALP activity assay and Fast Blue RR salt stain showed that 100 Hz could accelerate cells differentiation. Compared to the control group, 100 Hz could promote cell proliferation. Furthermore, 1 Hz to 10 Hz could improve calcium deposition in the intracellular matrix. Overall, these results indicate that 100Hz ES exhibits superior potentialities in osteogenesis, which should be beneficial for the clinical applications of ES for the treatment of bone diseases. PMID:27149625

  14. Enhanced metabolism of halogenated hydrocarbons in transgenic plants containing mammalian cytochrome P450 2E1

    NASA Astrophysics Data System (ADS)

    Lafferty Doty, Sharon; Shang, Tanya Q.; Wilson, Angela M.; Tangen, Jeff; Westergreen, Aram D.; Newman, Lee A.; Strand, Stuart E.; Gordon, Milton P.

    2000-06-01

    Chlorinated solvents, especially trichloroethylene (TCE), are the most widespread groundwater contaminants in the United States. Existing methods of pumping and treating are expensive and laborious. Phytoremediation, the use of plants for remediation of soil and groundwater pollution, is less expensive and has low maintenance; however, it requires large land areas and there are a limited number of suitable plants that are known to combine adaptation to a particular environment with efficient metabolism of the contaminant. In this work, we have engineered plants with a profound increase in metabolism of the most common contaminant, TCE, by introducing the mammalian cytochrome P450 2E1. This enzyme oxidizes a wide range of important pollutants, including TCE, ethylene dibromide, carbon tetrachloride, chloroform, and vinyl chloride. The transgenic plants had a dramatic enhancement in metabolism of TCE of up to 640-fold as compared with null vector control plants. The transgenic plants also showed an increased uptake and debromination of ethylene dibromide. Therefore, transgenic plants with this enzyme could be used for more efficient remediation of many sites contaminated with halogenated hydrocarbons.

  15. Rates of E1, E2, M1, and M2 transitions in Ni II

    NASA Astrophysics Data System (ADS)

    Cassidy, C. M.; Hibbert, A.; Ramsbottom, C. A.

    2016-03-01

    Aims: We present rates for all E1, E2, M1, and M2 transitions among the 295 fine-structure levels of the configurations 3d9, 3d84s, 3d74s2, 3d84p, and 3d74s4p, determined through an extensive configuration interaction calculation. Methods: The CIV3 code developed by Hibbert and coworkers is used to determine for these levels configuration interaction wave functions with relativistic effects introduced through the Breit-Pauli approximation. Results: Two different sets of calculations have been undertaken with different 3d and 4d functions to ascertain the effect of such variation. The main body of the text includes a representative selection of data, chosen so that key points can be discussed. Some analysis to assess the accuracy of the present data has been undertaken, including comparison with earlier calculations and the more limited range of experimental determinations. The full set of transition data is given in the supplementary material as it is very extensive. Conclusions: We believe that the present transition data are the best currently available. Full Table 4 and Tables 5-8 are only available at the CDS via anonymous ftp to http://cdsarc.u-strasbg.fr (ftp://130.79.128.5) or via http://cdsarc.u-strasbg.fr/viz-bin/qcat?J/A+A/587/A107

  16. Prostaglandin E1 effects on resting and cholinergically stimulated lower esophageal sphincter pressure in cats.

    PubMed

    Sinar, D R; Fletcher, J R; Castell, D O

    1981-04-01

    Intraluminal esophageal manometry with a sleeve catheter was used to compare the magnitude of decrease in lower esophageal sphincter (LES) pressure produced by an arterial or venous infusion of prostaglandin E1 in cats. Arterial PGE1 produced significantly lower LES pressures than venous PGE1 (p less than 0.05). Maximal decrease of 75% in basal LES pressure occurred with an associated 15% decrease in systolic blood pressure. The site of action of PGE1 in producing LES hypotension was studied by injection of either edrophonium, or bethanechol during the maximal PGE1 effect. Bethanechol, which acts directly on sphincteric smooth muscle, produced an increase in LES pressure during both saline and PGE1 infusion, while the increases in LES pressure seen with edrophonium during saline infusion were blocked during the PGE1 infusion. From these studies, we conclude that PGE1 produces LES hypotension in the cat by an inhibitory effect on the cholinergic pathway responsible for maintaining LES tone. These studies pharmacologically reproduce the LES pressure abnormality previously reported in the cat during acid-induced esophagitis and support the hypothesis that PGE1 may be involved in the pathogenesis of acute acid-induced lower esophageal sphincter abnormalities. PMID:6114513

  17. Decreased esophageal peristaltic amplitude in response to prostaglandin E1 and prostacyclin in the baboon.

    PubMed

    Sinar, D R; Cordova, C M; Fletcher, J R; Castell, D O

    1982-12-01

    Prostaglandins have been shown to produce significant decreases in lower esophageal sphincter pressure (LESP), although their effect on esophageal peristalsis is unknown. We studied the effect of infusion of prostaglandin E1 (PGE1) or prostacyclin (PI) on esophageal peristalsis in the proximal and distal esophagus in the awake baboon. Peristalsis was recorded using a polyvinyl catheter and a pneumohydraulic perfusion system and was induced by wet swallows. PGE1 infusion significantly (P less than 0.01) diminished peristaltic amplitude in proximal and distal esophagus by 51% and 77%, respectively. The wave duration was significantly (P less than 0.001) shortened by PGE1 in the distal esophagus, but not in the proximal esophagus. Similarly, prostacyclin significantly (P less than 0.05) decreased peristaltic amplitude in proximal and distal esophagus by 31% and 67%, respectively. As seen with PGE1, PI decreased distal esophageal amplitude significantly (P less than 0.02) more than proximal esophageal amplitude. Equivalent decreases in mean arterial blood pressure seen during prostaglandin infusion were reproduced by bleeding with no changes in measurements of peristaltic activity. Decreased peristaltic wave amplitude and duration suggest that prostaglandins exert a modulating local effect on esophageal muscle. In addition, this effect appears to be more pronounced on distal smooth muscle than on proximal striated muscle in the baboon esophagus. PMID:6756831

  18. Block adjustment of Chang'E-1 images based on rational function model

    NASA Astrophysics Data System (ADS)

    Liu, Bin; Liu, Yiliang; Di, Kaichang; Sun, Xiliang

    2014-05-01

    Chang'E-1(CE-1) is the first lunar orbiter of China's lunar exploration program. The CCD camera carried by CE-1 has acquired stereo images covering the entire lunar surface. Block adjustment and 3D mapping using CE-1 images are of great importance for morphological and other scientific research of the Moon. Traditional block adjustment based on rigorous sensor model is complicated due to a large number of parameters and possible correlations among them. To tackle this problem, this paper presents a block adjustment method using Rational Function Model (RFM). The RFM parameters are generated based on rigorous sensor model using virtual grid of control points. Afterwards, the RFM based block adjustment solves the refinement parameters through a least squares solution. Experimental results using CE-1 images located in Sinus Irdium show that the RFM can fit the rigorous sensor model with a high precision of 1% pixel level. Through the RFM-based block adjustment, the back-projection residuals in image space can be reduced from around 1.5 pixels to sub-pixel., indicating that RFM can replace rigorous sensor model for geometric processing of lunar images.

  19. Two-quasiparticle isomer, E1 hindrances and residual interactions in {sup 172}Tm

    SciTech Connect

    Hughes, R. O.; Lane, G. J.; Dracoulis, G. D.; Kibedi, T.; Nieminen, P.; Watanabe, H.

    2008-04-15

    The structure of the neutron-rich nucleus {sup 172}Tm has been studied using incomplete fusion of {sup 7}Li on an {sup 170}Er target at 30 MeV. A 190-{mu}s isomer at an excitation energy of 476 keV was identified using chopped beams and {gamma}-ray spectroscopy. The isomer decays with very inhibited E1 transitions to the rotational bands based on the parallel and antiparallel couplings of the {nu}5/2{sup -}[512] x {pi}1/2{sup +}[411] configuration, the latter (K{sup {pi}}=2{sup -}) being the ground state. The isomeric state has been assigned J{sup {pi}}=6{sup +}, arising from the energetically favored (parallel) coupling of the {nu}5/2{sup -}[512] x {pi}7/2{sup -}[523] configuration. The proton-neutron residual interaction was deduced for the configuration of the isomeric state and is found to agree with previous empirical studies.

  20. Cytoplasmic pH influences cytoplasmic calcium in MC3T3-E1 osteoblast cells

    NASA Technical Reports Server (NTRS)

    Lin, H. S.; Hughes-Fulford, M.; Kumegawa, M.; Pitts, A. C.; Snowdowne, K. W.

    1993-01-01

    We found that the cytoplasmic concentration of calcium (Cai) of MC3T3-E1 osteoblasts was influenced by the type of pH buffer we used in the perfusing medium, suggesting that intracellular pH (pHi) might influence Cai. To study this effect, the Cai and pHi were monitored as we applied various experimental conditions known to change pHi. Exposure to NH4Cl caused a transient increase in both pHi and Cai without a change in extracellular pH (pHo). Decreasing pHo and pHi by lowering the bicarbonate concentration of the medium decreased Cai, and increasing pHi by the removal of 5% CO2 increased Cai. Clamping pHi to known values with 10 microM nigericin, a potassium proton ionophore, also influenced Cai: acid pHi lowered Cai, whereas alkaline pHi increased it. The rise in Cai appears to be very sensitive to the extracellular concentration of calcium, suggesting the existence of a pH-sensitive calcium influx mechanism. We conclude that physiologic changes in pH could modulate Cai by controlling the influx of calcium ions and could change the time course of the Cai transient associated with hormonal activation.

  1. Mineralization initiation of MC3T3-E1 preosteoblast is suppressed under simulated microgravity condition.

    PubMed

    Hu, Li-fang; Li, Jing-bao; Qian, Ai-rong; Wang, Fei; Shang, Peng

    2015-04-01

    Microgravity decreases the differentiation of osteoblast. However, as this process is multistage and complex, the mechanism by which microgravity inhibits osteoblast differentiation is still unclear. We have previously found that 24 h acute treatment of simulated microgravity (SM) with a random positioning machine (RPM) significantly inhibited the differentiation of preosteoblasts and have explored whether osteoblasts show different response to microgravity condition at other stages, such as the mineralizing-stage. Murine MC3T3-E1 preosteoblasts induced for osteogenic differentiation for seven days were cultured either under normal gravity or SM conditions for 24 h. SM treatment significantly suppressed mineralized nodule formation. Alkaline phosphatase (ALP) activity was dramatically decreased, and the expression of ALP gene was downregulated. Expression of well-known markers and regulators for osteoblasts differentiation, including osteocalcin (OC), type I collagen α1 (Col Iα1), dentin matrix protein 1 (DMP1) and runt-related transcription factor 2 (Runx2), were downregulated. Western blot analysis showed that the phosphorylated extracellular signal-regulated kinase (p-ERK) level was lower under SM condition. Thus, the initiation of osteoblast mineralization is suppressed by SM condition, and the suppression may be through the regulation of ALP activity and the osteogenic gene expression. ERK signaling might be involved in this process. These results are relevant to the decrease of osteoblast maturation and bone formation under microgravity condition. PMID:25318973

  2. MC3T3-E1 Cell Response to Pure Titanium, Zirconia and Nano-Hydroxyapatite

    NASA Astrophysics Data System (ADS)

    Lee, Dong-Hwan; Han, Jung-Suk; Yang, Jae-Ho; Lee, Jai-Bong; Kim, Dae-Joon

    Titanium, zirconia and HAp were known as good biocompatible materials for tissue engineering. Osteblastic cell response is influence by the surface topography of material and its chemical composition as well. To evaluate the influence of different chemical compositions on osteoblast-like cells the specimens were polished until they have almost identical surface roughness. The commercially pure titanium, zirconia/alumina composite and nano-sized hydroxyapatite (HAp) specimens synthesized by hydrothermal method were used to evaluate the cell attachment, proliferation and differentiation. Confocal laser microscopy was used measurement of surface roughness, and flourescence microscopy and SEM were used to evaluate initial cell attachment and morphology after 3 hours. MTS assay was performed for cell proliferation after 1, 3, 7 days and ALP assay was used for cell differentiation after 7, 10, 14 days of cell culture period. Surface topography of nano-HAp specimen was almost identical compared with those of titanium and zirconia specimen. Under this condition, proliferation and differentiation of MC3T3-E1 cells was not significantly different with those on titanium and zirconia specimen. However, cells on Nano-HAp specimen showed quicker and more active cellular reaction for attachment when measured by the expression of adhesion proteins through confocal laser microscopy. The results suggested that the new nano-sized HAp can be applied as a suitable material for skeletal tissue engineering.

  3. 3D visualization of solar wind ion data from the Chang'E-1 exploration

    NASA Astrophysics Data System (ADS)

    Zhang, Tian; Sun, Yankui; Tang, Zesheng

    2011-10-01

    Chang'E-1 (abbreviation CE-1), China's first Moon-orbiting spacecraft launched in 2007, carried equipment called the Solar Wind Ion Detector (abbreviation SWID), which sent back tens of gigabytes of solar wind ion differential number flux data. These data are essential for furthering our understanding of the cislunar space environment. However, to fully comprehend and analyze these data presents considerable difficulties, not only because of their huge size (57 GB), but also because of their complexity. Therefore, a new 3D visualization method is developed to give a more intuitive representation than traditional 1D and 2D visualizations, and in particular to offer a better indication of the direction of the incident ion differential number flux and the relative spatial position of CE-1 with respect to the Sun, the Earth, and the Moon. First, a coordinate system named Selenocentric Solar Ecliptic (SSE) which is more suitable for our goal is chosen, and solar wind ion differential number flux vectors in SSE are calculated from Geocentric Solar Ecliptic System (GSE) and Moon Center Coordinate (MCC) coordinates of the spacecraft, and then the ion differential number flux distribution in SSE is visualized in 3D space. This visualization method is integrated into an interactive visualization analysis software tool named vtSWIDs, developed in MATLAB, which enables researchers to browse through numerous records and manipulate the visualization results in real time. The tool also provides some useful statistical analysis functions, and can be easily expanded.

  4. Characterization and application of monoclonal antibodies against Mycoplasma hyorhinis pyruvate dehydrogenase E1 complex subunit alpha.

    PubMed

    Chen, Dongjie; Wei, Yanwu; Huang, Liping; Wang, Yiping; Du, Wenjuan; Sun, Jianhui; Wu, Hongli; Feng, Li; Liu, Changming

    2016-04-01

    Mycoplasma hyorhinis is commonly found in the respiratory tract of pigs and is the etiological agent of polyserositis. The metabolic enzymes of M. hyorhinis may play important roles in host-pathogen interactions. We immunized BALB/c mice with sodium deoxycholate-extracted antigens (DOC-Ags) and screened 10 hybridomas that secreted antibodies against various M. hyorhinis proteins. Pyruvate dehydrogenase E1 complex subunit alpha (PDHA) was identified as the protein that reacted with five of the 10 monoclonal antibodies (mAbs). Sequence analysis indicated that PDHA was highly conserved among M. hyorhinis strains, but not among other mycoplasmas. We predicted the three-dimensional structure of PDHA and identified three epitopes ((277)RTEEEEK(283), (299)KDKKYITDE(307), and (350)LKEQKQHAKDY(360)). The mAb 1H12 we generated was used to detect M. hyorhinis PDHA in vitro and in piglets infected with M. hyorhinis. We observed that PDHA was mainly located in the epithelial cells of the lungs. Our results indicate that the mAbs we generated could be used to further investigate the structure and function of M. hyorhinis PDHA. In addition, they could be used in the differential diagnosis of M. hyorhinis and other mycoplasmas. PMID:26743652

  5. Hierarchical polymeric scaffolds support the growth of MC3T3-E1 cells.

    PubMed

    Akbarzadeh, Rosa; Minton, Joshua A; Janney, Cara S; Smith, Tyler A; James, Paul F; Yousefi, Azizeh-Mitra

    2015-02-01

    Tissue engineering makes use of the principles of biology and engineering to sustain 3D cell growth and promote tissue repair and/or regeneration. In this study, macro/microporous scaffold architectures have been developed using a hybrid solid freeform fabrication/thermally induced phase separation (TIPS) technique. Poly(lactic-co-glycolic acid) (PLGA) dissolved in 1,4-dioxane was used to generate a microporous matrix by the TIPS method. The 3D-bioplotting technique was used to fabricate 3D macroporous constructs made of polyethylene glycol (PEG). Embedding the PEG constructs inside the PLGA solution prior to the TIPS process and subsequent extraction of PEG following solvent removal (1,4-dioaxane) resulted in a macro/microporous structure. These hierarchical scaffolds with a bimodal pore size distribution (<50 and >300 μm) contained orthogonally interconnected macro-channels generated by the extracted PEG. The diameter of the macro-channels was varied by tuning the dispensing parameters of the 3D bioplotter. The in vitro cell culture using murine MC3T3-E1 cell line for 21 days demonstrated that these scaffolds could provide a favorable environment to support cell adhesion and growth. PMID:25665851

  6. Hybrid aligned nematic based measurement of the sum (e1+e3) of the flexoelectric coefficients

    NASA Astrophysics Data System (ADS)

    Tartan, Chloe C.; Elston, Steve J.

    2015-02-01

    A new method has been established for the measurement of the sum of the flexoelectric coefficients e1+e3 in liquid crystals by exploiting the properties of highly ionic materials in order to screen out the internal bias due to the different surface alignment polarities in a Hybrid Aligned Nematic (HAN) liquid crystal device. It has been shown that responses to pulses are independent of the external offset of a signal applied to a HAN device filled with a highly ionic material. Driving the device with step changes in the offset leads to either a transient increase or transient decrease in the response, depending on the polarity of the offset, while the equilibrium response remains the same. The time constant of the transient effect is consistent with the relaxation time of the ions present in the material. Assuming these ions screen out the internal bias completely, the remaining response can be used as a measure of the flexoelectric effect. Based on this approach, a value of (10 ± 2) pC m-1 was found for the modulus of the flexoelectric sum in the standard commercial eutectic E70 nematic liquid crystal mixture.

  7. Ziram Causes Dopaminergic Cell Damage by Inhibiting E1 Ligase of the Proteasome*

    PubMed Central

    Chou, Arthur P.; Maidment, Nigel; Klintenberg, Rebecka; Casida, John E.; Li, Sharon; Fitzmaurice, Arthur G.; Fernagut, Pierre-Olivier; Mortazavi, Farzad; Chesselet, Marie-Francoise; Bronstein, Jeff M.

    2008-01-01

    The etiology of Parkinson disease (PD) is unclear but may involve environmental toxins such as pesticides leading to dysfunction of the ubiquitin proteasome system (UPS). Here, we measured the relative toxicity of ziram (a UPS inhibitor) and analogs to dopaminergic neurons and examined the mechanism of cell death. UPS (26 S) activity was measured in cell lines after exposure to ziram and related compounds. Dimethyl- and diethyldithiocarbamates including ziram were potent UPS inhibitors. Primary ventral mesencephalic cultures were exposed to ziram, and cell toxicity was assessed by staining for tyrosine hydroxylase (TH) and NeuN antigen. Ziram caused a preferential damage to TH+ neurons and elevated α-synuclein levels but did not increase aggregate formation. Mechanistically, ziram altered UPS function through interfering with the targeting of substrates by inhibiting ubiquitin E1 ligase. Sodium dimethyldithiocarbamate administered to mice for 2 weeks resulted in persistent motor deficits and a mild reduction in striatal TH staining but no nigral cell loss. These results demonstrate that ziram causes selective dopaminergic cell damage in vitro by inhibiting an important degradative pathway implicated in the etiology of PD. Chronic exposure to widely used dithiocarbamate fungicides may contribute to the development of PD, and elucidation of its mechanism would identify a new potential therapeutic target. PMID:18818210

  8. Lunar absolute reflectance as observed by Chang'E-1 Imaging Interferometer

    NASA Astrophysics Data System (ADS)

    Zhang, Jiang; Ling, ZongCheng; Liu, JianZhong; Wu, ZhongChen; Li, Bo; Ni, YuHeng

    2015-08-01

    Lunar absolute reflectance, which describes the fraction of solar radiation reflected by the Moon, is fundamental for the Chang'E-1 Imaging Interferometer (IIM) to map lunar mineralogical and elemental distributions. Recent observations made by the Spectral Irradiance Monitor (SIM) onboard the Solar Radiation and Climate Experiment (SORCE) spacecraft indicate that temporal variation in the solar radiation might have non-negligible influence on reflectance calculation, and the SIM measurements are different from the two previously used solar irradiances, i.e., ATLAS3 and Newkur. To provide reliable science results, we examined solar irradiance variability with the SIM daily observations, derived lunar absolute reflectances from the IIM 2A radiance with the SIM, ATLAS3 and Newkur data, and compared them with the Chandrayaan-1 Moon Mineralogy Mapper (M3), the Robotic Lunar Observatory (ROLO) and the Kaguya Multispectral Imager (MI) results. The temporal variability of the SIM solar irradiance is 0.25%-1.1% in the IIM spectral range, and less than 0.2% during the IIM observations. Nevertheless, the differences between the SIM measurements and the ATLAS3 and Newkur data can respectively rise up to 8% and 5% at particular IIM bands, resulting in discrepancy between which might affect compositional mapping. The IIM absolute reflectance we derived for the Moon using the SIM data, except for the last two bands, is consistent with the ROLO and the MI observations, although it is lower.

  9. Strength of anchor bolts in masonry

    NASA Astrophysics Data System (ADS)

    Whitlock, A. R.; Brown, R. H.

    1983-08-01

    The strength properties of bent bar bolts in masonry were evaluated. The variables studied include bolt size, loading direction, embedment substrate, and bolt location. It is found that bolt strength increases with bolt size up to the point where failure is controlled by masonry strength. The loading direction, substrate, and location of bolts are shown to be significant factors in the strength of anchors. Previously developed empirical and analytical design equations for anchor bolts in reinforced concrete and masonry are investigated and compared to the study results. The equations are found to accurately predict the mean strengths of anchor bolts.

  10. Choosing a strength training program for kids.

    PubMed

    Benjamin, Holly J; Glow, Kimberly M; Mees, Patricia D

    2003-09-01

    Whether your child is involved in sports, does recreational activities, or just needs to be more active, a strength training program can be one part of a well-balanced youth fitness program. Improved muscle coordination gained from strength training can increase athletic performance and help prevent some on-field injuries in sports. You may notice your child gaining more self confidence and better social skills along with muscle strength. Good strength training can improve bone health and also help overweight kids lose unwanted pounds. You want your child's exercise to be safe, but what should you look for before your child begins strength training? PMID:20086489

  11. Strength of Welded Aircraft Joints

    NASA Technical Reports Server (NTRS)

    Brueggeman, W C

    1937-01-01

    This investigation is a continuation of work started in 1928 and described in NACA-TR-348 which shows that the insertion of gusset plates was the most satisfactory way of strengthening a joint. Additional tests of the present series show that joints of this type could be improved by cutting out the portion of the plate between the intersecting tubes. T and lattice joints in thin-walled tubing 1 1/2 by 0.020 inch have somewhat lower strengths than joints in tubing of greater wall thickness because of failure by local buckling. In welding the thin-walled tubing, the recently developed "carburizing flux" process was found to be the only method capable of producing joints free from cracks. The "magnetic powder" inspection was used to detect cracks in the joints and flaws in the tubing.

  12. Cobalt: for strength and color

    USGS Publications Warehouse

    Boland, Maeve A.; Kropschot, S.J.

    2011-01-01

    Cobalt is a shiny, gray, brittle metal that is best known for creating an intense blue color in glass and paints. It is frequently used in the manufacture of rechargeable batteries and to create alloys that maintain their strength at high temperatures. It is also one of the essential trace elements (or "micronutrients") that humans and many other living creatures require for good health. Cobalt is an important component in many aerospace, defense, and medical applications and is a key element in many clean energy technologies. The name cobalt comes from the German word kobold, meaning goblin. It was given this name by medieval miners who believed that troublesome goblins replaced the valuable metals in their ore with a substance that emitted poisonous fumes when smelted. The Swedish chemist Georg Brandt isolated metallic cobalt-the first new metal to be discovered since ancient times-in about 1735 and identified some of its valuable properties.

  13. Strain rate, temperature, and humidity on strength and moduli of a graphite/epoxy composite

    NASA Technical Reports Server (NTRS)

    Lifshitz, J. M.

    1981-01-01

    Results of an experimental study of the influence of strain rate, temperature and humidity on the mechanical behavior of a graphite/epoxy fiber composite are presented. Three principal strengths (longitudinal, transverse and shear) and four basic moduli (E1, E2, G12 and U12) of a unidirectional graphite/epoxy composite were followed as a function of strain rate, temperature and humidity. Each test was performed at a constant tensile strain rate in an environmental chamber providing simultaneous temperature and humidity control. Prior to testing, specimens were given a moisture preconditioning treatment at 60 C. Values for the matrix dominated moduli and strength were significantly influenced by both environmental and rate parameters, whereas the fiber dominated moduli were not. However, the longitudinal strength was significantly influenced by temperature and moisture content. A qualitative explanation for these observations is presented.

  14. Preparation, mechanical strengths, and thermal

    NASA Astrophysics Data System (ADS)

    Inoue, A.; Furukawa, S.; Hagiwara, M.; Masumoto, T.

    1987-05-01

    Ni-based amorphous wires with good bending ductility have been prepared for Ni75Si8B17 and Ni78P12B10 alloys containing 1 to 2 at. pct Al or Zr by melt spinning in rotating water. The enhancement of the wire-formation tendency by the addition of Al has been clarified to be due to the increase in the stability of the melt jet through the formation of a thin A12O3 film on the outer surface. The maximum wire diameter is about 190 to 200 μm for the Ni-Si (or P)-B-Al alloys and increases to about 250 μm for the Ni-Si-B-Al-Cr alloys containing 4 to 6 at. pct Cr. The tensile fracture strength and fracture elongation are 2730 MPa and 2.9 pct for (Ni0.75Si0.08B0.17 99Al1) wire and 2170 MPa and 2.4 pct for (Ni0.78P0.12B0.1)99Al1 wire. These wires exhibit a fatigue limit under dynamic bending strain in air with a relative humidity of 65 pct; this limit is 0.50 pct for a Ni-Si-B-Al wire, which is higher by 0.15 pct than that of a Fe75Si10B15 amorphous wire. Furthermore, the Ni-base wires do not fracture during a 180-deg bending even for a sample annealed at temperatures just below the crystallization temperature, in sharp contrast to high embrittlement tendency for Fe-base amorphous alloys. Thus, the Ni-based amorphous wires have been shown to be an attractive material similar to Fe- and Co-based amorphous wires because of its high static and dynamic strength, high ductility, high stability to thermal embrittlement, and good corrosion resistance.

  15. Trabecular bone strength at the knee.

    PubMed

    Hvid, I

    1988-02-01

    The axial strength of trabecular bone at the knee is critical for the maintenance of support and fixation of the prosthetic components after total surface knee arthroplasty. The resistance of trabecular bone to penetration was measured posteriorly, centrally, and anteriorly in each of the tibial and femoral condyles in 150 consecutive total knee arthroplasties. Forty-seven rheumatoid knees and 88 osteoarthritic knees were evaluated. The correlation of bone strength with selected clinical parameters was found to be too poor to predict bone strength. Tibial bone strength was lower in rheumatoid than in osteoarthritic knees. Steroid medication did not influence tibial bone strength in rheumatoid arthritis. The distribution of bone strength between the medial and lateral condyles was closely dependent on knee alignment, with high medial strength in varus knees. At the unloaded condyle, strength was reduced relative to the findings for normally aligned knees. At the tibia, strength decreased with depth from the resection surface, while at the femur the converse was true. Tibial bone strength, both condylar and overall average, was lower than values reported in studies of normal cadaver knees. Evaluation of the absolute bone strength at the tibial condyles suggested that the values too low to meet load-bearing requirements after well-aligned knee replacement were infrequent. PMID:3276421

  16. Adhesive strength of autologous fibrin glue.

    PubMed

    Yoshida, H; Hirozane, K; Kamiya, A

    2000-03-01

    To establish an easy and rapid method for measuring the adhesive strength of fibrin glue and to clarify the factor(s) most affecting the strength, a study was made on the effect of the concentration of plasma components on the strength of cryoprecipitate (Cryo) prepared from a subject's own autologous plasma to be used as fibrin glue. The adhesive strength of the Cryo was measured with various supporting materials instead of animal skin using a tester of tension and compression. The results were as follows: (1) the strength of Cryo applied to ground flat glass (4 cm2) was significantly greater than that applied to clear glass, clear plastic, or smooth and flat wood chips; (2) the adhesive strength of Cryo depended on the concentration of thrombin with the optimal concentration being 50 units/ml; (3) the concentration of CaCl2 did not affect the adhesive strength of Cryo; (4) the adhesive reaction was dependent on the temperature and the adhesive strength more quickly reached a steady state at 37 degrees C than at lower temperature; (5) the adhesive strength was correlated well with the total concentration of fibrinogen and fibronectin. These results indicate that the adhesive strength of Cryo can be easily and quickly evaluated using a tester and ground glass with thrombin at 50 units/ml, and that the adhesive strength of Cryo can be predicted from the total concentration of fibrinogen and fibronectin. PMID:10726885

  17. The Distribution of Subjective Memory Strength: List Strength and Response Bias

    ERIC Educational Resources Information Center

    Criss, Amy H.

    2009-01-01

    Models of recognition memory assume that memory decisions are based partially on the subjective strength of the test item. Models agree that the subjective strength of targets increases with additional time for encoding however the origin of the subjective strength of foils remains disputed. Under the fixed strength assumption the distribution of

  18. The Strengths Assessment Inventory: Reliability of a New Measure of Psychosocial Strengths for Youth

    ERIC Educational Resources Information Center

    Brazeau, James N.; Teatero, Missy L.; Rawana, Edward P.; Brownlee, Keith; Blanchette, Loretta R.

    2012-01-01

    A new measure, the Strengths Assessment Inventory-Youth self-report (SAI-Y), was recently developed to assess the strengths of children and adolescents between the ages of 10 and 18 years. The SAI-Y differs from similar measures in that it provides a comprehensive assessment of strengths that are intrinsic to the individual as well as strengths

  19. The Distribution of Subjective Memory Strength: List Strength and Response Bias

    ERIC Educational Resources Information Center

    Criss, Amy H.

    2009-01-01

    Models of recognition memory assume that memory decisions are based partially on the subjective strength of the test item. Models agree that the subjective strength of targets increases with additional time for encoding however the origin of the subjective strength of foils remains disputed. Under the fixed strength assumption the distribution of…

  20. Molecular and enzymatic characterization of alkaline lipase from Bacillus amyloliquefaciens E1PA isolated from lipid-rich food waste.

    PubMed

    Saengsanga, Thanakorn; Siripornadulsil, Wilailak; Siripornadulsil, Surasak

    2016-01-01

    Bacillus amyloliquefaciens E1PA is a lipase-producing strain that was originally isolated from lipid-rich food waste, and the production of its lipase was found to be induced by vegetable oils. The E1PA lipase was successfully expressed and secreted in a heterologous Escherichia coli host and was ultimately purified. The conserved pentapeptide motif Ala-His-Ser-Met-Gly was observed at positions 108-112. The purified recombinant lipase was stable over a pH range of 4.0-11.0 at 40 °C and exhibited maximal activity at pH 10. The recombinant E1PA lipase hydrolyzed a wide range of acyl esters (C4-C18). However, the highest activity (3.5 units mg(-1)) was observed when the p-nitrophenyl ester of myristate (C14) was used as a substrate. Compared to the lipases produced by Bacillus spp., the E1PA lipase displayed a structural molecular mass excluding the leader sequence (19.22 kDa) and a pI (9.82) that were similar to those reported for B. amyloliquefaciens lipases and lipase subfamily I.4 but that were quite distinct from those of lipase subfamily I.5 (approximately 43 kDa, pI 6). These results suggested that Bacillus lipases are closely related. Although the recombinant E1PA lipase digested only certain oils, the wild-type E1PA lipase degraded a variety of oils, including blended and re-used cooking oils. The recombinant and wild-type forms of the E1PA lipase were able to digest heterogeneous lipid-rich food waste at similar levels; this result suggests that this lipase can function even when it solely consists of its structural enzyme component. The enzyme exhibited lipid hydrolysis ability as either an intracellular domain of the recombinant protein or an extracellular domain secreted by the E1PA strain. However, the recombinant lipase showed higher activity than the wild-type E1PA lipase, indicating that the recombinant protein from E. coli possessed effective lipase activity. Thus, the inducible alkaline E1PA lipase exhibited the ability to act on a broad spectrum of substrates, and the effective form produced in the heterogeneous host can be further developed for several applications, such as biodiesel production and lipase production. PMID:26672445

  1. Breit-Pauli energy levels belonging to 2p 4, 2s2p 5, 2p 6, 2p 33ℓ configurations and all E1 transitions among these levels in Mg V

    NASA Astrophysics Data System (ADS)

    Deb, N. C.; Hibbert, A.

    2007-07-01

    We present accurate oscillator strengths, line strengths and radiative rates for 1073 E1 transitions among the 86 levels belonging to 2s 22p 4, 2s2p 5, 2p 6, and 2s 22p 3( 4S o, 2D o, 2P o)3ℓ configurations in Mg V. We have used 1s and 2s Hartree-Fock orbitals, re-optimized 2p on 2p 3( 2D o)3s 3D o and optimized 3s,3p,3d orbitals on real states. Sixteen additional orbitals up to 8d are optimized either as a correction to n = 3 physical orbitals or as a correlation orbital. A very large set of configurations including up to three electron promotions are used to account for all important correlation effects. All of the main five terms in the Breit-Pauli operator (except the orbit-orbit interaction) are included in order to account for the relativistic effects. Small adjustments to the diagonal elements of the Hamiltonian matrix are made to bring the calculated energies within a few cm -1 of the corresponding NIST recommended data wherever available. The calculated oscillator strengths, line strengths, and radiative rates for almost all of the E1 transitions show excellent agreement with the corresponding MCDF results of Fischer. The recent results of Bhatia et al. are found to be consistently higher by 20-45%. The accuracy of the present calculation is considered to be better than the NIST accuracy ratings for various transitions.

  2. Absolute Calibration of the Chang’E-1 IIM camera and its preliminary application

    NASA Astrophysics Data System (ADS)

    Wu, Y.

    2009-12-01

    The Chinese first lunar satellite, Chang’E-1, was launched on 24 Oct 2007 and ended its 16-month mission by impacting the Moon on 1st Mar 2009. The Interference Imaging spectrometer (IIM) which covers wavelengths from 480 to 960 nm with 32 channels onboard Chang’E-1 is aimed to acquire the composition of the Moon. The reflectance spectra of Apollo 16 soil 62231 measured in the laboratory and the corresponding area of undisturbed soil of Apollo 16 site was used for the calibration of IIM data. The sensor response is inhomogeneous and hence caused the hue changing from left to right. The area covering the calibration site was selected for the correction of the shading of IIM data. Two factors ensured that the corrected reflectance of each pixel has the physical meaning, i.e., reflectance of the surface. Firstly, the area for selecting the correction standard line is the same place as the land surface used for calibration, i.e., Apollo 16 landing site area. Secondly, the correction factor is derived from fitting the reflectance of the correction standard line which is corrected to the reflectance of 62231. The Aristarchus crater is one of the most concerned craters on the Moon due to its geological complex and spectral and geochemical diversity. In this preliminary study we chose the Aristarchus crater as our first example. Some individual pixels with high albedo and low reflectance at 918 nm intersperse among the west wall, which suggests that some individual fresh surface may be exposed. From its dispersed distribution and fresh surface we conclude that the landslide may exist on the west wall. It also seems that there are some collapsed materials on the left floor on the Lunar Orbit photograph. We found that a high correlation exists between the absorption center of the mafic minerals and the wavelength at which the first derivative equals to 0. The key parameter to identify minerals, absorption center, was produced with this correlation. The absorption center of the northwest wall is the shortest, while the south wall is the longest among the walls. The 8-km Väisälä crater also exhibits the lithologic heterogeneity. The absorption center for Väisälä crater is dichotomal while for Aristarchus is trisectional. The mineral assemblages of the southwest and northeast wall of Väisälä are probable orthopyroxene and clinopyroxene, respectively. Besides the two kinds of pyroxene, the south wall of Aristarchus probably contains olivine. A region with shorter absorption center than that of the dark mantle deposits was found between the Aristarchus and Väisälä crater. The absorption center of this area is similar with that of the northeast wall of Väisälä but longer than that of the nearest wall (southwest wall) of Väisälä. The origin of the materials of this region needs further investigating.

  3. Anik-E1 and E2 satellite failures of January 1994 revisited

    NASA Astrophysics Data System (ADS)

    Lam, H.-L.; Boteler, D. H.; Burlton, B.; Evans, J.

    2012-10-01

    The consecutive failures of the geosynchronous Anik-E1 communication satellite on January 20, 1994, and Anik-E2 about nine hours later on January 21 (both incidents occurred on January 20 local time) received considerable publicity because the malfunctions of the satellites disrupted television and computer data transmissions across Canada, as well as telephone services to remote northern communities for hours. This often-cited event is revisited here with materials not covered before. Using publicly available information, Anik-E failure details, media coverage, recovery effort and cost incurred are first presented. This is then followed by scrutiny of space weather conditions pertinent to the occurrences of the Anik-E upsets. We trace the space weather episode's inception on the Sun, propagation through interplanetary medium, and manifestation in magnetic field variations as well as in energetic electron flux increases, and its eventual impact on the Anik-Es. The genesis of the energetic electron enhancements that have been blamed for the satellite malfunctions is thus traceable via high-speed solar wind stream with Alfven wave fluctuations to a longitudinally wide coronal hole on the Sun. Furthermore, strong magnetic pulsations preceding electron flux peaks indicate Pc5 ULF (Ultra Low Frequency) waves as a probable acceleration mechanism for the energetic electron flux enhancement that resulted in the internal charging of the Anik-Es. The magnetic fluctuations may even be possible triggers for the subsequent discharge that caused the satellites to malfunction. This incident illustrates that satellite operators should be on alert for elevated high-energy electron environment that is above established thresholds, as specifications in satellite design may not render a satellite immune from internal charging.

  4. Development, Optimization, and Characterization of PEGylated Nanoemulsion of Prostaglandin E1 for Long Circulation.

    PubMed

    Cheng, Ying; Liu, Miao; Hu, Huijing; Liu, Daozhou; Zhou, Siyuan

    2016-04-01

    Lipo-PGE1 is the most widely used formulation of PGE1 in clinic. However, PGE1 is easier to leak out from lipo-PGE1 and this will lead to the phlebophlogosis when intravenous injection. The stability of lipo-PGE1 in storage and in vivo is also discounted. The aim of this study is to develop a long-circulating prostaglandin E1-loaded nanoemulsion modified with 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG) to improve the stability and pharmacokinetics profiles of lipo-PGE1. PEGylated PGE1 nanoemulsion was prepared using a dispersing-homogenized method. The stability of nanoemulsion in 1 month was investigated. Pharmacokinetic studies were employed to evaluate the in vivo profile of the optimized nanoemulsion. The optimized nanoemulsion PGE1-PEG2000(1%)-NE showed an oil droplet size <100 nm with a surface charge of -14 mV. Approximately, 97% of the PGE1 was encapsulated in the nanoemulsion. The particle size, zeta potential, and drug loading of PGE1-PEG2000(1%)-NE were stable in 1 month. After PGE1-PEG2000(1%)-NE was intravenously administered to rats, the area under curve (AUC) and half-life of PGE1 were, respectively, 1.47-fold and 5.98-fold higher than those of lipo-PGE1 (commercial formulation). PGE1-PEG2000(1%)-NE was an ideal formulation for prolonging the elimination time of PGE1. This novel parenteral colloidal delivery system of PGE1 has a promising potential in clinic use. PMID:26195071

  5. Loss of platelet alpha 2-adrenergic receptors during simulated extracorporeal circulation: prevention with prostaglandin E1

    SciTech Connect

    Wachtogel, Y.T.; Musial, J.; Jenkin, B.; Niewiarowski, S.; Edmunds, L.H. Jr.; Colman, R.W.

    1985-05-01

    Cardiopulmonary bypass prolongs bleeding time and increases postoperative blood loss. During in vitro recirculation in an extracorporeal circuit containing a membrane oxygenator and primed with fresh heparinized human blood, the authors previously observed thrombocytopenia, impaired platelet aggregation, and depletion of granular contents, all of which were prevented with prostaglandin E1 (PGE1). To investigate these changes further, they studied the number and affinity of platelet alpha 2-adrenergic receptors by measuring the binding of /sup 3/H-yohimbine. Before recirculation, they found 235 alpha 2-adrenergic receptors per platelet, a Kd of 3.37 nmol/L, complete aggregation with 1.04 mumol/L epinephrine, and a platelet count of 281,000 microliters/sup -1/. After 2 minutes of recirculation, 9.44 mumol/L epinephrine was required to produce complete aggregation, and the platelet count was 104,000 microliters-1 (44% of control). After 2 hours of recirculation, the platelet count had increased to 123,000 microliters/sup -1/. However, epinephrine did not induce platelet aggregation even at 100 mumol/L. Moreover, alpha 2-adrenergic binding sites were not detectable, and affinity for yohimbine could not be calculated. Two minutes after PGE1 0.3 mumol/L was added to the circuit, platelet numbers, response to epinephrine, alpha 2-adrenergic binding sites per platelet, and affinity for yohimbine were not significantly different from control values. At 2 hours, the number of alpha 2-adrenergic sites was not significantly changed from control, but the affinity of yohimbine for platelets was significantly decreased 2.5-fold.

  6. Global analysis of protein-protein interactions reveals multiple CYP2E1-reductase complexes.

    PubMed

    Jamakhandi, Arvind P; Kuzmic, Petr; Sanders, Daniel E; Miller, Grover P

    2007-09-01

    Although a single binary functional complex between cytochrome P450 (P450 or CYP for a specific isoform) and cytochrome P450 reductase (CPR) has been generally accepted in the literature, this simple model failed to explain the experimentally observed catalytic activity of recombinant CYP2E1 in dependence on the total concentration of the added CPR-K56Q mutant. Our rejection of the simplest 1:1 binding model was based on two independent lines of experimental evidence. First, under the assumption of the 1:1 binding model, separate analyses of titration curves obtained while varying either P450 or CPR concentrations individually produced contradictory results. Second, an asymmetric Job plot suggested the existence of higher order molecular complexes. To identify the most probable complexation mechanism, we generated a comprehensive data set where the concentrations of both P450 and P450 were varied simultaneously, rather than one at a time. The resulting two-dimensional data were globally fit to 32 candidate mechanistic models, involving the formation of binary, ternary, and quaternary P450.CPR complexes, in the absence or presence or P450 and CPR homodimers. Of the 32 candidate models (mechanisms), two models were approximately equally successful in explaining our experimental data. The first plausible model involves the binary complex P450.CPR, the quaternary complex (P450)2.(CPR)2, and the homodimer (P450)2. The second plausible model additionally involves a weakly bound ternary complex (P450)2.CPR. Importantly, only the binary complex P450.CPR seems catalytically active in either of the two most probable mechanisms. PMID:17685587

  7. Characterization of the Self-Cleaving Effector Protein NopE1 of Bradyrhizobium japonicum ▿

    PubMed Central

    Schirrmeister, Jana; Friedrich, Lars; Wenzel, Mandy; Hoppe, Markus; Wolf, Christine; Göttfert, Michael; Zehner, Susanne

    2011-01-01

    NopE1 is a type III-secreted protein of the symbiont Bradyrhizobium japonicum which is expressed in nodules. In vitro it exhibits self-cleavage in a duplicated domain of unknown function (DUF1521) but only in the presence of calcium. Here we show that either domain is self-sufficient for cleavage. An exchange of the aspartic acid residue at the cleavage site with asparagine prevented cleavage; however, cleavage was still observed with glutamic acid at the same position, indicating that a negative charge at the cleavage site is sufficient. Close to each cleavage site, an EF-hand-like motif is present. A replacement of one of the conserved aspartic acid residues with alanine prevented cleavage at the neighboring site. Except for EDTA, none of several protease inhibitors blocked cleavage, suggesting that a known protease-like mechanism is not involved in the reaction. In line with this, the reaction takes place within a broad pH and temperature range. Interestingly, magnesium, manganese, and several other divalent cations did not induce cleavage, indicating a highly specific calcium-binding site. Based on results obtained by blue-native gel electrophoresis, it is likely that the uncleaved protein forms a dimer and that the fragments of the cleaved protein oligomerize. A database search reveals that the DUF1521 domain is present in proteins encoded by Burkholderia phytofirmans PsNJ (a plant growth-promoting betaproteobacterium) and Vibrio coralliilyticus ATCC BAA450 (a pathogenic gammaproteobacterium). Obviously, this domain is more widespread in proteobacteria, and it might contribute to the interaction with hosts. PMID:21642459

  8. The Human Orphan Nuclear Receptor Tailless (TLX, NR2E1) Is Druggable

    PubMed Central

    Benod, Cindy; Villagomez, Rosa; Filgueira, Carly S.; Hwang, Peter K.; Leonard, Paul G.; Poncet-Montange, Guillaume; Rajagopalan, Senapathy; Fletterick, Robert J.; Gustafsson, Jan-ke; Webb, Paul

    2014-01-01

    Nuclear receptors (NRs) are an important group of ligand-dependent transcriptional factors. Presently, no natural or synthetic ligand has been identified for a large group of orphan NRs. Small molecules to target these orphan NRs will provide unique resources for uncovering regulatory systems that impact human health and to modulate these pathways with drugs. The orphan NR tailless (TLX, NR2E1), a transcriptional repressor, is a major player in neurogenesis and Neural Stem Cell (NSC) derived brain tumors. No chemical probes that modulate TLX activity are available, and it is not clear whether TLX is druggable. To assess TLX ligand binding capacity, we created homology models of the TLX ligand binding domain (LBD). Results suggest that TLX belongs to an emerging class of NRs that lack LBD helices ?1 and ?2 and that it has potential to form a large open ligand binding pocket (LBP). Using a medium throughput screening strategy, we investigated direct binding of 20,000 compounds to purified human TLX protein and verified interactions with a secondary (orthogonal) assay. We then assessed effects of verified binders on TLX activity using luciferase assays. As a result, we report identification of three compounds (ccrp1, ccrp2 and ccrp3) that bind to recombinant TLX protein with affinities in the high nanomolar to low micromolar range and enhance TLX transcriptional repressive activity. We conclude that TLX is druggable and propose that our lead compounds could serve as scaffolds to derive more potent ligands. While our ligands potentiate TLX repressive activity, the question of whether it is possible to develop ligands to de-repress TLX activity remains open. PMID:24936658

  9. Genetic dissection of the transactivating domain of the E1a 289R protein of adenovirus type 2.

    PubMed Central

    Fahnestock, M L; Lewis, J B

    1989-01-01

    A series of linker-scanning, deletion, and frameshift mutations were made in the pm975 variant of the adenovirus type 2 E1a gene, which expresses only the larger of the two major E1a proteins. Most of these were within the 46-amino-acid segment unique to the larger E1a protein product (the 289R protein), which confers on it the ability to activate in trans the expression of other genes. The mutations were recombined into virus and assayed by in vitro transcription in nuclei isolated from infected cells for their ability to activate the transcription of other viral early genes and of the endogenous hsp70 gene. Mutant E1a proteins from which the 289R-unique segment was removed by deletion or truncation did not completely lose the ability to transactivate by comparison with a virus which makes no E1a at all, indicating that sequences outside this domain are active in the positive regulation of transcription. The E1a mutations tested fell into several classes: those that increased transactivation of virtually all genes, those that severely depressed transactivation of all genes, and those that depressed transactivation only moderately. Each mutation had similar effects on the expression of all transcription units tested, indicating a common process in their transactivation. However, some mutants in the third category decreased transactivation of some induced genes more severely than of others. Such gene-specific defects suggest the existence of subclasses of E1a-responsive transcription units, consistent with the involvement of diverse proteins in the transactivation of different genes. Two specific structural components of the transactivating domain, a putative metal-binding element and a region with high potential for beta-sheet formation at its carboxy-terminus, appear to be important to the transactivation function. Images PMID:2522557

  10. Expression, purification and guanine nucleotide binding characterization of Arabidopsis RabE1d13-185 GTPase.

    PubMed

    Gao, Yi; Tan, Lingling; Dong, Chun-Hai; Hou, Xiaomin

    2016-03-01

    Arabidopsis RabE1d subclass plays important plant-specific functions in plant growth and development, response to ethylene and defence to plant pathogen, besides their basic cellular role in membrane trafficking. In this study, we present the expression, purification, and characterization of the recombinant core domain of AtRabE1d13-185. AtRabE1d13-185 was successfully expressed in Escherichia coli and purified via two-step nickel affinity chromatography followed by gel filtration, and identified single band in SDS-PAGE. The resultant protein was functionally active, as determined by interaction with guanine nucleotide by a fluorescence-based assay. The intrinsic tryptophan of AtRabE1d13-185 showed fluorescence resonance energy transfer (FRET) effect upon forming complex with fluorescent methylanthraniloyl (mant)-GDP, but quenched when binding with non-labelled guanine nucleotide. The association rate of mantGDP with AtRabE1d13-185 was determined to be 3.48 × 10(7) s(-1) M(-1). The dissociation rates of GDP and mantGDP from the complex with AtRabE1d13-185 were similar. The koff values were determined to be 4.02 × 10(-4) s(-1) based on the FRET effect for the AtRabE1d13-185:GDP and 5.41 × 10(-4) s(-1) for mantGDP excited directly. PMID:26611608

  11. Immunological Characterization of a Modified Vaccinia Virus Ankara Vector Expressing the Human Papillomavirus 16 E1 Protein

    PubMed Central

    Remy-Ziller, Christelle; Germain, Claire; Spindler, Anita; Hoffmann, Chantal; Silvestre, Nathalie; Rooke, Ronald; Bonnefoy, Jean-Yves

    2014-01-01

    Women showing normal cytology but diagnosed with a persistent high-risk human papillomavirus (HR-HPV) infection have a higher risk of developing high-grade cervical intraepithelial neoplasia and cervical cancer than noninfected women. As no therapeutic management other than surveillance is offered to these women, there is a major challenge to develop novel targeted therapies dedicated to the treatment of these patients. As such, E1 and E2 antigens, expressed early in the HPV life cycle, represent very interesting candidates. Both proteins are necessary for maintaining coordinated viral replication and gene synthesis during the differentiation process of the epithelium and are essential for the virus to complete its normal and propagative replication cycle. In the present study, we evaluated a new active targeted immunotherapeutic, a modified vaccinia virus Ankara (MVA) vector containing the E1 sequence of HPV16, aimed at inducing cellular immune responses with the potential to help and clear persistent HPV16-related infection. We carried out an extensive comparative time course analysis of the cellular immune responses induced by different schedules of immunization in C57BL/6 mice. We showed that multiple injections of MVA-E1 allowed sustained HPV16 E1-specific cellular immune responses in vaccinated mice and had no impact on the exhaustion phenotype of the generated HPV16 E1-specific CD8+ T cells, but they led to the differentiation of multifunctional effector T cells with high cytotoxic capacity. This study provides proof of concept that an MVA expressing HPV16 E1 can induce robust and long-lasting E1-specific responses and warrants further development of this candidate. PMID:24307238

  12. Nitric Oxide-Induced Autophagy in MC3T3-E1 Cells is Associated with Cytoprotection via AMPK Activation

    PubMed Central

    Yang, Jung Yoon; Park, Min Young; Park, Sam Young; Yoo, Hong Il; Kim, Min Seok; Kim, Jae Hyung

    2015-01-01

    Nitric oxide (NO) is important in the regulation of bone remodeling, whereas high concentration of NO promotes cell death of osteoblast. However, it is not clear yet whether NO-induced autophagy is implicated in cell death or survival of osteoblast. The present study is aimed to examine the role of NO-induced autophagy in the MC3T3-E1 cells and their underlying molecular mechanism. The effect of sodium nitroprusside (SNP), an NO donor, on the cytotoxicity of the MC3T3-E1 cells was determined by MTT assay and expression of apoptosis or autophagy associated molecules was evaluated by western blot analysis. The morphological observation of autophagy and apoptosis by acridine orange stain and TUNEL assay were performed, respectively. Treatment of SNP decreased the cell viability of the MC3T3-E1 cells in dose- and time-dependent manner. SNP increased expression levels of p62, ATG7, Beclin-1 and LC3-II, as typical autophagic markers and augmented acidic autophagolysosomal vacuoles, detected by acridine orange staining. However, pretreatment with 3-methyladenine (3MA), the specific inhibitor for autophagy, decreased cell viability, whereas increased the cleavage of PARP and caspase-3 in the SNP-treated MC3T3-E1 cells. AMP-activated protein kinase (AMPK), a major autophagy regulatory kinase, was activated in SNP-treated MC3T3-E1 cells. In addition, pretreatment with compound C, an inhibitor of AMPK, decreased cell viability, whereas increased the number of apoptotic cells, cleaved PARP and caspase-3 levels compared to those of SNP-treated MC3T3-E1 cells. Taken together, it is speculated that NO-induced autophagy functions as a survival mechanism via AMPK activation against apoptosis in the MC3T3-E1 cells. PMID:26557017

  13. Solution Structure, Copper Binding and Backbone Dynamics of Recombinant Ber e 1–The Major Allergen from Brazil Nut

    PubMed Central

    Rundqvist, Louise; Tengel, Tobias; Zdunek, Janusz; Björn, Erik; Schleucher, Jürgen; Alcocer, Marcos J. C.; Larsson, Göran

    2012-01-01

    Background The 2S albumin Ber e 1 is the major allergen in Brazil nuts. Previous findings indicated that the protein alone does not cause an allergenic response in mice, but the addition of components from a Brazil nut lipid fraction were required. Structural details of Ber e 1 may contribute to the understanding of the allergenic properties of the protein and its potential interaction partners. Methodology/Principal Findings The solution structure of recombinant Ber e 1 was solved using NMR spectroscopy and measurements of the protein back bone dynamics at a residue-specific level were extracted using 15N-spin relaxation. A hydrophobic cavity was identified in the structure of Ber e 1. Using the paramagnetic relaxation enhancement property of Cu2+ in conjunction with NMR, it was shown that Ber e 1 is able to specifically interact with the divalent copper ion and the binding site was modeled into the structure. The IgE binding region as well as the copper binding site show increased dynamics on both fast ps-ns timescale as well as slower µs-ms timescale. Conclusions/Significance The overall fold of Ber e 1 is similar to other 2S albumins, but the hydrophobic cavity resembles that of a homologous non-specific lipid transfer protein. Ber e 1 is the first 2S albumin shown to interact with Cu2+ ions. This Cu2+ binding has minimal effect on the electrostatic potential on the surface of the protein, but the charge distribution within the hydrophobic cavity is significantly altered. As the hydrophobic cavity is likely to be involved in a putative lipid interaction the Cu2+ can in turn affect the interaction that is essential to provoke an allergenic response. PMID:23056307

  14. Yin Yang 1 Negatively Regulates the Differentiation-Specific E1 Promoter of Human Papillomavirus Type 6

    PubMed Central

    Ai, Wandong; Narahari, Janaki; Roman, Ann

    2000-01-01

    Human papillomavirus type 6 (HPV-6) is a low-risk HPV whose replication cycle, like that of all HPVs, is differentiation dependent. We have previously shown that CCAAT displacement protein (CDP) binds the differentiation-induced HPV-6 E1 promoter and negatively regulates its activity in undifferentiated cells (W. Ai, E. Toussaint, and A. Roman, J. Virol. 73:4220–4229, 1999). Using electrophoretic mobility shift assays (EMSAs), we now report that Yin Yang 1 (YY1), a multifunctional protein that can act as a transcriptional activator or repressor and that can also inhibit HPV replication in vitro, binds the HPV-6 E1 promoter. EMSAs, using subfragments of the promoter as competitors, showed that the YY1 binding site is located at the 5′ end of the E1 promoter. When a putative YY1 site was mutated, the ability of YY1 to bind was greatly decreased. The activity of the mutated E1 promoter, monitored with the reporter gene luciferase, was threefold greater than that of the wild-type promoter, suggesting that YY1 negatively regulates HPV-6 E1 promoter activity. Nuclear extracts from differentiated keratinocytes showed decreased binding of YY1 to the wild-type promoter. Consistent with this, in differentiated keratinocytes, the activity of the transfected luciferase gene transcribed from the mutated promoter was comparable to that of the wild-type promoter; both promoters were up-regulated in differentiated keratinocytes compared to undifferentiated cells. These data suggest that YY1 functions in undifferentiated keratinocytes but not in differentiated keratinocytes. Both the wild-type and mutated promoters could be negatively regulated by overexpression of a plasmid encoding CDP. Thus, both YY1 and CDP appear to be negative regulators of the differentiation-induced HPV-6 E1 promoter and thereby the HPV life cycle. In contrast, only binding of CDP was detected using the E1 promoter of the high-risk HPV-31. PMID:10799595

  15. Natural variation in the genes responsible for maturity loci E1, E2, E3 and E4 in soybean

    PubMed Central

    Tsubokura, Yasutaka; Watanabe, Satoshi; Xia, Zhengjun; Kanamori, Hiroyuki; Yamagata, Harumi; Kaga, Akito; Katayose, Yuichi; Abe, Jun; Ishimoto, Masao; Harada, Kyuya

    2014-01-01

    Background and Aims The timing of flowering has a direct impact on successful seed production in plants. Flowering of soybean (Glycine max) is controlled by several E loci, and previous studies identified the genes responsible for the flowering loci E1, E2, E3 and E4. However, natural variation in these genes has not been fully elucidated. The aims of this study were the identification of new alleles, establishment of allele diagnoses, examination of allelic combinations for adaptability, and analysis of the integrated effect of these loci on flowering. Methods The sequences of these genes and their flanking regions were determined for 39 accessions by primer walking. Systematic discrimination among alleles was performed using DNA markers. Genotypes at the E1E4 loci were determined for 63 accessions covering several ecological types using DNA markers and sequencing, and flowering times of these accessions at three sowing times were recorded. Key Results A new allele with an insertion of a long interspersed nuclear element (LINE) at the promoter of the E1 locus (e1-re) was identified. Insertion and deletion of 36 bases in the eighth intron (E2-in and E2-dl) were observed at the E2 locus. Systematic discrimination among the alleles at the E1E3 loci was achieved using PCR-based markers. Allelic combinations at the E1E4 loci were found to be associated with ecological types, and about 6266 % of variation of flowering time could be attributed to these loci. Conclusions The study advances understanding of the combined roles of the E1E4 loci in flowering and geographic adaptation, and suggests the existence of unidentified genes for flowering in soybean, PMID:24284817

  16. Proliferation and osteogenic response of MC3T3-E1 pre-osteoblastic cells on porous zirconia ceramics stabilized with magnesia or yttria.

    PubMed

    Hadjicharalambous, Chrystalleni; Mygdali, Evdokia; Prymak, Oleg; Buyakov, Ales; Kulkov, Sergei; Chatzinikolaidou, Maria

    2015-11-01

    Dense zirconia ceramics are used in bone applications due to their mechanical strength and biocompatibility, but lack osseointegration. A porous interface in contact with bone tissue may lead to better bone bonding but the biological properties of porous zirconia are not widely explored. The present study focuses on the manufacturing of an yttria- (YSZ) and a magnesia-stabilized (MgSZ) porous zirconia, and on their in vitro biological investigation. The sintered ceramics had similar characteristics of porosity, pore size and interconnectivity. Their elastic moduli and compressive strength values were within the range of the values of human cortical bone. MC3T3-E1 pre-osteoblasts were used to investigate the proliferation, alkaline phosphatase (ALP) activity, collagen deposition and expression profile of four genes involved in bone metabolism of cells on porous ceramics. Scanning electron and fluorescence microscopy were employed to visualize cell morphology and growth. Pre-osteoblasts adhered well on both ceramics but cell numbers on YSZ were higher. Cells exhibited an increase in ALP activity and collagen deposition after 14 days on both MgSZ and YSZ, with higher levels on YSZ. Real-time quantitative polymerase chain reaction (qPCR) showed that the expression of bone sialoprotein (Bsp) and collagen type I (col1aI) were significantly higher on YSZ. No significant differences were found in their ability to regulate the early gene expression of Runx2 and Alp. Nevertheless, the biomineralized calcium content was similar on both ceramics after 21 days, indicating that despite chemical differences, both scaffolds direct the pre-osteoblasts toward a mature state capable of mineralizing the extracellular matrix. PMID:25847599

  17. Roles of Cytochrome P4502E1 Gene Polymorphisms and the Risks of Alcoholic Liver Disease: A Meta-Analysis

    PubMed Central

    Zeng, Tao; Guo, Fang-Fang; Zhang, Cui-Li; Song, Fu-Yong; Zhao, Xiu-Lan; Xie, Ke-Qin

    2013-01-01

    Background Previous studies investigating the association between cytochrome P4502E1 (CYP2E1) polymorphisms and the risk of alcoholic liver diseases (ALD) have yielded conflicting results. Thus, a meta-analysis was performed to clarify the association between CYP2E1 polymorphisms and the risks of ALD. Methods A comprehensive literature search was conducted to identify the relevant studies. The fixed or random effect model was selected based on the heterogeneity test among studies. Publication bias was estimated using Beggs funnel plots and Eggers regression test. Results A total of 27 and 9 studies were finally included for the association between the CYP2E1 Pst I/Rsa I or Dra I polymorphisms and the risks of ALD, respectively. Overall, the combined results showed that homozygous genotype c2c2 was significantly associated with increase risk of ALD in worldwide populations (c2c2 vs. c1c1: OR?=?3.12, 95%CI 1.915.11) when ALD patients were compared with alcoholics without ALD. Significant associations between CYP2E1 Pst I/Rsa I polymorphism and ALD risk were also observed in Asians (c2c2 vs. c1c1: OR?=?4.11, 95%CI 2.327.29) and in Caucasians (c2c2/c1c2 vs. c1c1: OR?=?1.58, 95%CI 1.042.42) when ALD patients were compared with alcoholics without ALD. However, subgroup analysis stratified by ALD types showed that CYP2E1 Pst I/Rsa I polymorphism was not significantly associated with the risks of alcoholic cirrhosis (ALC). No significant association was observed between CYP2E1 Dra I polymorphism and ALD risks. Conclusion This meta-analysis suggested that CYP2E1 Pst I/Rsa I polymorphism might be not significantly associated with advanced form of ALD (ALC), but might be significantly associated with other form of ALD such as steatosis, hepatisis, fibrosis. Furthermore, CYP2E1 Dra I polymorphism might be not significantly associated with the ALD risks. Since potential confounders could not be ruled out completely, further studies were needed to confirm these results. PMID:23335995

  18. Mouse osteoblastic cell line (MC3T3-E1) expresses extracellular calcium (Ca2+o)-sensing receptor and its agonists stimulate chemotaxis and proliferation of MC3T3-E1 cells

    NASA Technical Reports Server (NTRS)

    Yamaguchi, T.; Chattopadhyay, N.; Kifor, O.; Butters, R. R. Jr; Sugimoto, T.; Brown, E. M.; O'Malley, B. W. (Principal Investigator)

    1998-01-01

    The calcium-sensing receptor (CaR) is a G protein-coupled receptor that plays key roles in extracellular calcium ion (Ca2+o) homeostasis in parathyroid gland and kidney. Osteoblasts appear at sites of osteoclastic bone resorption during bone remodeling in the "reversal" phase following osteoclastic resorption and preceding bone formation. Bone resorption produces substantial local increases in Ca2+o that could provide a signal for osteoblasts in the vicinity, leading us to determine whether such osteoblasts express the CaR. In this study, we used the mouse osteoblastic, clonal cell line MC3T3-E1. Both immunocytochemistry and Western blot analysis, using an antiserum specific for the CaR, detected CaR protein in MC3T3-E1 cells. We also identified CaR transcripts in MC3T3-E1 cells by Northern analysis using a CaR-specific riboprobe and by reverse transcription-polymerase chain reaction with CaR-specific primers, followed by nucleotide sequencing of the amplified products. Exposure of MC3T3-E1 cells to high Ca2+o (up to 4.8 mM) or the polycationic CaR agonists, neomycin and gadolinium (Gd3+), stimulated both chemotaxis and DNA synthesis in MC3T3-E1 cells. Therefore, taken together, our data strongly suggest that the osteoblastic cell line MC3T3-E1 possesses both CaR protein and mRNA very similar, if not identical, to those in parathyroid and kidney. Furthermore, the CaR in these osteoblasts could play a key role in regulating bone turnover by stimulating the proliferation and migration of such cells to sites of bone resorption as a result of local release of Ca2+o.

  19. Ubiquitin-specific protease 7 is a regulator of ubiquitin-conjugating enzyme UbE2E1.

    PubMed

    Sarkari, Feroz; Wheaton, Keith; La Delfa, Anthony; Mohamed, Majda; Shaikh, Faryal; Khatun, Rahima; Arrowsmith, Cheryl H; Frappier, Lori; Saridakis, Vivian; Sheng, Yi

    2013-06-01

    Ubiquitin-specific protease 7 (USP7) is a deubiquitinating enzyme found in all eukaryotes that catalyzes the removal of ubiquitin from specific target proteins. Here, we report that UbE2E1, an E2 ubiquitin conjugation enzyme with a unique N-terminal extension, is a novel USP7-interacting protein. USP7 forms a complex with UbE2E1 in vitro and in vivo through the ASTS USP7 binding motif within its N-terminal extension in an identical manner with other known USP7 binding proteins. We show that USP7 attenuates UbE2E1-mediated ubiquitination, an effect that requires the N-terminal ASTS sequence of UbE2E1 as well as the catalytic activity of USP7. Additionally, USP7 is critical in maintaining the steady state levels of UbE2E1 in cells. This study reveals a new cellular mechanism that couples the opposing activities of the ubiquitination machinery and a deubiquitinating enzyme to maintain and modulate the dynamic balance of the ubiquitin-proteasome system. PMID:23603909

  20. Endophytic Bacillus subtilis Strain E1R-J Is a Promising Biocontrol Agent for Wheat Powdery Mildew

    PubMed Central

    Gong, Yufei; Huo, Yunxia; Han, Qingmei; Kang, Zhensheng; Huang, Lili

    2015-01-01

    In this study, the biocontrol efficacies of 14 endophytic bacterial strains were tested against Blumeria graminis f. sp. tritici (Bgt) in pot experiments under greenhouse conditions. Bacillus subtilis strain E1R-j significantly reduced disease index and exhibited the best control (90.97%). When different formulations of E1R-j were sprayed 24 h before Bgt inoculation, fermentation liquid without bacterial cell and crude protein suspension displayed the similar effects; and they reduced disease index more than bacterial cell suspension (109 cfu mL−1) and fermentation liquid without protein. The control effects were not significantly different between 1011 and 109 cfu mL−1 of bacterial cell suspension but were higher than 107 cfu mL−1. Further observations showed that conidial germination and appressorial formation of Bgt were retarded by spraying E1R-j 24 h before Bgt inoculation. Compared with the water check, conidial germination and appressorial formation were decreased by 43.3% and 42.7%, respectively. In the treatment with E1R-j, the number of houstoria significantly reduced and the speed of mycelial extension was slowed down in the wheat leaves. Scanning electron microscopy observation revealed that E1R-j significantly suppressed the conidial germination and caused rupture and deformation of germ tubes. On the surface of wheat leaves, mycelia and conidiophores became shrinking. PMID:25759819