Science.gov

Sample records for e1 two-phonon strengths

  1. Fragmentation of two-phonon {gamma}-vibrational strength in deformed nuclei

    SciTech Connect

    Wu, C.Y.; Cline, D.

    1996-12-31

    Rotational and vibrational modes of collective motion. are very useful in classifying the low-lying excited states in deformed nuclei. The rotational mode of collective motion is characterized by rotational bands having correlated level energies and strongly-enhanced E2 matrix elements. The lowest intrinsic excitation with I,K{sup {pi}} = 2,2{sup +} in even-even deformed nuclei, typically occurring at {approx}1 MeV, is classified as a one-phonon {gamma}-vibration state. In a pure harmonic vibration limit, the expected two-phonon {gamma}-vibration states with I,K{sup {pi}} = 0,0{sup +} and 4,4{sup +} should have excitation energies at twice that of the I,K{sup {pi}} = 2,2{sup +} excitation, i.e. {approx}2 MeV, which usually is above the pairing gap leading to possible mixing with two-quasiparticle configurations. Therefore, the question of the localization of two-phonon {gamma}-vibration strength has been raised because mixing may lead to fragmentation of the two-phonon strength over a range of excitation energy. For several well-deformed nuclei, an assignment of I,K{sup {pi}}=4,4{sup +} states as being two-phonon vibrational excitations has been suggested based on the excitation energies and the predominant {gamma}-ray decay to the I,K{sup {pi}}=2,2{sup +} state. However, absolute B(E2) values connecting the presumed two- and one-phonon states are the only unambiguous measure of double phonon excitation. Such B(E2) data are available for {sup 156}Gd, {sup 160}Dy, {sup 168}Er, {sup 232}Th, and {sup 186,188,190,192}Os. Except for {sup 160}Dy, the measured B(E2) values range from 2-3 Weisskopf units in {sup 156}Gd to 10-20 Weisskopf units in osmium nuclei; enhancement that is consistent with collective modes of motion.

  2. 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.

  3. E1 and M1 γ-strength functions in 144Nd

    DOE PAGES

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

    2015-12-14

    Both E1 and M1 γ-strength functions below the neutron separation energy were analyzed based on experimental data from 143Nd(n,γ)144Nd and 143Nd(n,γα)140Ce reactions. It is confirmed that the commonly adopted E1 model based on the temperature dependence of the width of the giant dipole resonance works well. The popular M1 strength function due to the spin-flip magnetic resonance located near the neutron binding energy is not capable of reproducing experimental data. As a result, the low-energy enhancement of the M1 strength or the energy-independent model of Weisskopf, both leading to the low-energy strength sizable to E1 one, fit experimental data best.

  4. E1 and M1 γ-strength functions in 144Nd

    SciTech Connect

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

    2015-12-14

    Both E1 and M1 γ-strength functions below the neutron separation energy were analyzed based on experimental data from 143Nd(n,γ)144Nd and 143Nd(n,γα)140Ce reactions. It is confirmed that the commonly adopted E1 model based on the temperature dependence of the width of the giant dipole resonance works well. The popular M1 strength function due to the spin-flip magnetic resonance located near the neutron binding energy is not capable of reproducing experimental data. As a result, the low-energy enhancement of the M1 strength or the energy-independent model of Weisskopf, both leading to the low-energy strength sizable to E1 one, fit experimental data best.

  5. Optomechanics with two-phonon driving

    NASA Astrophysics Data System (ADS)

    Levitan, B. A.; Metelmann, A.; Clerk, A. A.

    2016-09-01

    We consider the physics of an optomechanical cavity subject to coherent two-phonon driving, i.e. degenerate parametric amplification of the mechanical mode. We show that in such a system, the cavity mode can effectively ‘inherit’ parametric driving from the mechanics, yielding phase-sensitive amplification and squeezing of optical signals reflected from the cavity. We also demonstrate how such a system can be used to perform single-quadrature detection of a near-resonant narrow-band force applied to the mechanics with extremely low added noise from the optics. The system also exhibits strong differences from a conventional degenerate parametric amplifier: in particular, the cavity spectral function can become negative, indicating a negative effective photon temperature.

  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. Low-lying E1,M1, and E2 strength distributions in Xe124,126,128,129,130,131,132,134,136: Systematic photon scattering experiments in the mass region of a nuclear shape or phase transition

    NASA Astrophysics Data System (ADS)

    von Garrel, H.; Brentano, P. Von; Fransen, C.; Friessner, G.; Hollmann, N.; Jolie, J.; Käppeler, F.; Käubler, L.; Kneissl, U.; Kohstall, C.; Kostov, L.; Linnemann, A.; Mücher, D.; Pietralla, N.; Pitz, H. H.; Rusev, G.; Scheck, M.; Schilling, K. D.; Scholl, C.; Schwengner, R.; Stedile, F.; Walter, S.; Werner, V.; Wisshak, K.

    2006-05-01

    Systematic nuclear resonance fluorescence (NRF) experiments on all nine stable (seven even-even and two odd-mass) Xe isotopes have been performed at the bremsstrahlung facility of the 4.3-MV Stuttgart Dynamitron accelerator. For the first time thin-walled, high-pressure gas targets (about 70 bar) with highly enriched target material were used in NRF experiments. Precise excitation energies, transition strengths, spins, and decay branching ratios were obtained for numerous states, most of them previously unknown. The systematics of the observed E1 two-phonon excitations (2+⊗3-) and M1 excitations to 1+ mixed-symmetry states in the even-even isotopes are discussed with respect to the new critical point symmetry E(5). The fragmentation of these fundamental dipole excitation modes in the odd-mass isotopes Xe129,131 is shown and discussed. In the even-even nuclei several low-lying E2 excitations were observed.

  8. Lifetimes of two-phonon 1 - states in even N = 82 nuclei

    NASA Astrophysics Data System (ADS)

    Herzberg, R.-D.; Bauske, I.; von Brentano, P.; Eckert, Th.; Fischer, R.; Geiger, W.; Kneissl, U.; Margraf, J.; Maser, H.; Pietralla, N.; Pitz, H. H.; Zilges, A.

    1995-02-01

    Photon scattering experiments on the semi magic nuclei 138Ba and 140Ce have been performed at the bremsstrahlung facility of the Stuttgart Dynamitron. Two-phonon 1 - states were excited at 4.026 and 3.643 MeV excitation energy respectively as well as a number of 2 + states between 2 and 4 MeV. Lifetimes and upper limits for the decay to other low-lying states were extracted from the data and are compared to lifetimes given in the literature. The systematics of B(E1) values and energies of the 1 - two-phonon states is analysed using additional data from previously published (γ, γ') experiments on the neighbouring isotones 142Nd and 144Sm.

  9. 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.

  10. Low-lying E1 and M1 strengths in the deformed nucleus 160Gd

    NASA Astrophysics Data System (ADS)

    Friedrichs, H.; Häger, D.; von Brentano, P.; Heil, R. D.; Herzberg, R.-D.; Kneissl, U.; Margraf, J.; Müller, G.; Pitz, H. H.; Schlitt, B.; Schumacher, M.; Wesselborg, C.; Zilges, A.

    1994-01-01

    Measurements of the linear polarization of resonantly scattered photons have been used for model-independent parity determinations in nuclear resonance fluorescence experiments. The experiments have been performed at the bremsstrahlung facility of the Stuttgart Dynamitron accelerator. The use of two different Compton polarimeters, a five Ge-detector arrangement and a sectored single-crystal Ge-polarimeter enabled the parity assignments. Positive parities were established for a cluster of states near 3.3 MeV (M1 transitions) to be ascribed to the so-called scissors mode. In addition an enhanced electric dipole excitation near 2.5 MeV (2471 keV, B(E1) ↑ = 3.0 ± 0.4 × 10 -3e2fm 2) has been observed as in the neighbouring isotopes 162Dy and 150Nd. These 1 - states systematically exhibit an uncommon decay branching hinting at K-mixing.

  11. Systematic study of low-lying E1 strength using the time-dependent mean field theory

    SciTech Connect

    Ebata, S.; Nakatsukasa, T.; Inakura, T.

    2012-11-12

    We carry out systematic investigation of electric dipole (E1) mode from light to heavy nuclei, using a new time-dependent mean field theory: the Canonical-basis Time-Dependent Hartree-Fock-Bogoliubov (Cb-TDHFB) theory. The Cb-TDHFB in the three-dimensional coordinate space representation can deal with pairing correlation and any kind of deformation in the timedependent framework. We report the neutron-number dependence of the low-energy E1 mode for light (A > 40) and heavy isotopes (A < 100) around N= 82.

  12. Reinvestigation of two-phonon γ-vibrational band in neutron-rich 114Pd

    NASA Astrophysics Data System (ADS)

    Huang, Y.; Zhu, S. J.; Hamilton, J. H.; Ramayya, A. V.; Wang, E. H.; Liu, Y. X.; Sun, Y.; Hwang, J. K.; Xiao, Z. G.; Li, H. J.; Luo, Y. X.; Rasmussen, J. O.; Ter-Akopian, G. M.; Oganessian, Yu. Ts.

    2016-08-01

    The level structure in neutron-rich 114Pd nucleus has been reinvestigated by measuring prompt γ rays emitted in the spontaneous fission of 252Cf. A two-phonon γ-vibrational band built on the 1639.3keV level is observed, which confirms the previous suggestion from a β-decay experiment. Systematical comparison supports the assignment for a two-phonon γ-vibrational band in 114Pd. Triaxial projected shell model calculations for the multi-phonon γ bands of 114Pd are in good agreement with the experimental data.

  13. A note on two-phonon coherent anti-stokes Raman scattering

    SciTech Connect

    Shen, Y. R.

    1981-01-01

    Difference-frequency mixing of two pump waves can in principle excite two coherent phonon waves via the parametric process. Finally, only when the phonon excitation is small can the nonlinear susceptibility of two-phonon coherent anti-Stokes Raman scattering be described as proportional to the product of two Raman tensors.

  14. 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.

  15. Two-dimensional image edge enhancement by two-phonon Bragg diffraction

    SciTech Connect

    Kotov, V M; Shkerdin, G N; Bulyuk, A N

    2011-12-31

    A model of acousto-optic interaction is developed for describing two-phonon Bragg diffraction in a gyrotropic single TeO{sub 2} crystal, which makes allowance for ellipticity of crystal optical eigenwaves. Formation of a two-dimensional edge of an image in the first diffraction order is explained by invoking a three-dimensional description for the wave surfaces of optical eigenwaves.

  16. Two-phonon processes of intraband relaxation in the terahertz regime in quantum dots.

    PubMed

    Wang, Zi-Wu; Li, Shu-Shen

    2011-06-01

    We theoretically investigate the intraband relaxation of quantum dots in the terahertz regime due to two acoustic phonon scattering by applying a lattice relaxation approach based on the deformation potential coupling between electrons and acoustic phonons. In particular, we find that the relaxation time depends strongly on the ratio of two acoustic phonons. The influences of the energy separation between the ground and first excited state, the quantum dot height, and the lattice temperature on the relaxation time are also discussed. Our theoretical results not only give a reasonable explanation for the current experimental measurement but also provide some insight into two-phonon intraband relaxation in quantum dots.

  17. Two-dimensional image edge enhancement in the two-phonon diffraction

    SciTech Connect

    Kotov, V M; Averin, S V; Shkerdin, G N; Voronko, A I

    2010-06-23

    We suggest using the two-phonon Bragg scattering regime for two-dimensional image edge enhancement by means of acousto-optic (AO) diffraction on a single sound wave. Image edge enhancement is demonstrated in the first diffraction order by using an AO cell made of the TeO{sub 2} single crystal. To explain this effect, a three-dimensional model of AO interaction is proposed, which takes into account the angular selectivity of diffraction both in the plane of Bragg scattering and in the plane orthogonal to it. (optical data processing)

  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. Hong-Ou-Mandel interference of two phonons in trapped ions.

    PubMed

    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.

  20. Search for the two-phonon octupole vibrational state in {sup 208}Pb

    SciTech Connect

    Blumenthal, D.J.; Henning, W.; Janssens, R.V.F.

    1995-08-01

    We performed an experiment to search for the two-phonon octupole vibrational state in {sup 208}Pb. Thick targets of {sup 208}Pb, {sup 209}Bi, {sup 58,64}Ni, and {sup 160}Gd were bombarded with 1305 MeV beams of were bombard {sup 208}Pb supplied by ATLAS. Gamma rays were detected using the Argonne-Notre Dame BGO gamma-ray facility, consisting of 12 Compton-suppressed germanium detectors surrounding an array of 50 BGO scintillators. We identified some 30 known gamma rays from {sup 208}Pb in the spectra gated by the 5{sup -} {yields} 3{sup -} and 3{sup -} {yields} 0{sup +} transitions in {sup 208}Pb. In addition, after unfolding these spectra for Compton response, we observed broad coincident structures in the energy region expected for the 2-phonon states. Furthermore, we confirmed the placement of a 2485 keV line observed previously in {sup 207}Pb and find no evidence consistent with the placement of this line in {sup 208}Pb. We are currently in the process of investigating the origin of the broadened lines observed in the spectra, extracting the excitation probability of states in {sup 208}Pb, and determining the relative probability of mutual excitation and neutron transfer in this reaction. An additional experiment is also being performed to collect much higher statistics germanium-germanium coincidence data for the thick {sup 208}Pb target.

  1. Dipole strength distributions in stable odd-mass nuclei in the vicinity of the N=82 isotones

    NASA Astrophysics Data System (ADS)

    Scheck, Marcus

    2006-10-01

    The low-lying dipole strength distributions in the odd-mass nuclei ^135Ba, ^137Ba, ^139La and ^141Pr were studied in nuclear resonance fluorescence (NRF) experiments performed at the Stuttgart Dynamitron facility. experiments used bremsstrahlung beams with endpoint energies of 4.1 MeV. The spin selective NRF reaction allowed the excitation of states through dipole transitions, up to 4 MeV. A special focus is the fragmented E1-strength of transitions connecting the ground state to states of the [2^+ 3^-] particle/hole coupling. The summed strength of the odd-mass nuclei is compared with the E1-strength of the [2^+ 3^-]1^- two-phonon states of the neighboring even-even core nuclei.

  2. 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.

  3. Fragmentation of low-lying dipole strength in the odd-mass nucleus 133Cs

    NASA Astrophysics Data System (ADS)

    Besserer, J.; Beck, O.; von Brentano, P.; Eckert, T.; Herzberg, R.-D.; Jäger, D.; Kneissl, U.; Margraf, J.; Maser, H.; Nord, A.; Pietralla, N.; Pitz, H. H.; Zilges, A.

    1997-09-01

    The fragmentation of low-lying dipole strength in the odd-mass nucleus 133Cs has been investigated in nuclear resonance fluorescence (NRF) experiments performed at the bremsstrahlung beam of the Stuttgart Dynamitron accelerator at an end-point energy of 4.1 MeV. In the excitation energy range 2.3 - 3.7 MeV in total 22 new dipole excitations were observed. From the high-resolution γ-ray spectra measured by three high-efficiency Ge detectors the reduced excitation probabilities B(E1)↑ or B(M1)↑ were deduced. The fragmentation and absolute total strengths of the detected dipole excitations are compared with results for the neighboring even-even, γ-soft nucleus 134Ba, where both, rather strong scissors mode-like M1 and two-phonon E1 excitations are known from recent NRF experiments.

  4. Dipole Strength Distributions in 124,126,128,130,132,134,136Xe:. a Systematic Study in the Mass Region of a Nuclear Shape Transition

    NASA Astrophysics Data System (ADS)

    Kneissl, U.

    2005-03-01

    Systematic nuclear resonance fluorescence experiments (NRF) on all 7 stable even-even Xe isotopes have been performed at the bremsstrahlung facility of the 4.3 MV Stuttgart Dynamitron accelerator. For the first time thin-walled, high-pressure gas targets (about 70 bar) were used in NRF experiments. Precise excitation energies, transition strengths, spins, and decay branching ratios were obtained for numerous states, most of them unknown so far. The systematics of the observed E1 two-phonon excitations (2+ ⊗ 3-) and M1 excitations to 1+ mixed symmetry states are discussed with respect to the new critical point symmetry E(5).

  5. 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.

  6. 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…

  7. 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.

  8. 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.

  9. Studying an advanced regime of the non-collinear two-phonon light scattering for applications to the optical spectrum analysis

    NASA Astrophysics Data System (ADS)

    Shcherbakov, Alexandre S.; Arellanes, Adan O.

    2016-03-01

    Principally new features of the non-collinear two-phonon light scattering governed by elastic waves of finite amplitude in birefringent bulk crystals are detected and observed. The main goals of our investigations are to reveal novel important details inherent in the nonlinearity of this effect and to study properties of similar parametric nonlinearity both theoretically and experimentally in wide-aperture crystals with moderate linear acoustic attenuation. An additional degree of freedom represented by the dispersive birefringence factor, which can be distinguished within this nonlinear phenomenon, is characterized. This physical degree of freedom gives us a one-of-a-kind opportunity to apply the strongly non-linear two-phonon light scattering in practice for the first time. The local unit-level maxima in the distribution of light scattered into the second order appear periodically as the acoustic power density grows. It makes possible to identify a few transfer function profiles peculiar to these maxima in the isolated planes of angular-frequency mismatches. These maxima give us an opportunity to choose the desirable profile for the transfer function at the fixed angle of incidence for the incoming light beam with a wide spectrum .The needed theoretical analysis is developed and proof-of-principle experiments, performed with a specially designed wide-aperture acousto-optical cell made of the calomel (α-Hg2Cl2) crystal, are presented. The obtained spectral resolution ~0.235 Å at 405 nm (i.e. the resolving power ~17,200) can be compared with the most advanced acousto-optical spectrometers for space/airborne operations. Evidently, our results with the calomel-based acousto-optical cell look like the best we can mention at the moment.

  10. Characterization of the non-collinear acousto-optical cell based on calomel (Hg2Cl2) crystal and operating within the two-phonon light scattering

    NASA Astrophysics Data System (ADS)

    Shcherbakov, Alexandre S.; Arellanes, Adan O.

    2016-03-01

    Performances of any system for data processing based on acousto-optical technique are mainly determined by parameters of the acousto-optical cell (AOC) exploited within the schematic arrangement. Here, basic properties of the AOC, involved into a novel processor for precise optical spectrum analysis dedicated to modern astrophysical applications, are considered. Because potential applications of this processor will be focused on investigations in extra-galactic astronomy as well as studies of extra-solar planets, an advanced regime of the non-collinear two-phonon light scattering has been elaborated for spectrum analysis with significantly improved spectral resolution. Under similar uprated requirements, the AOC, based on that specific regime in the calomel (Hg2Cl2) crystal, had been chosen, and its parameters were analyzed theoretically and verified experimentally. Then, the adequate approach to estimating the frequency/spectral bandwidth and spectral resolution had been developed. The bandwidth was calculated and experimentally realized with the additionally involved tilt angle of light incidence, allowing variations for acoustic frequencies. The resolution was characterized taking into account its doubling peculiar to the nonlinear two-phonon mechanism of light scattering. Proof-of-principle experiments were performed with the calomel AOC of 52 mm optical aperture, providing ~94% efficiency in the transmitted light due to the slow-shear acoustic mode of finite amplitude (the acoustic power density ~150 mW/mm2) with the velocity of 0.347×105 cm/s at the radio-wave acoustic frequency ~71 MHz. As a result, we have obtained the spectral resolution <0.235 Å within the spectral bandwidth <290 Å that looks as the best one can mention at the moment in acousto-optics.

  11. 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.

  12. 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.

  13. Shell-model description of E1 excitation

    NASA Astrophysics Data System (ADS)

    Shimizu, Noritaka; Utsuno, Yutaka; Togashi, Tomoaki; Otsuka, Takaharu; Honma, Michio

    2014-09-01

    We discuss a microscopic description of E1 excitations based on shell-model calculations. We performed large-scale shell-model calculations for Ca isotopes with Lanczos-strength-function method and sd - pf - sdg model space allowing up to 3 ℏω excitation and obtained their photoabsorption cross sections. It gives a very good description of giant dipole and low-lying pygmy resonances rather independently of smoothing parameter. We also present the feasiblity of the Monte Carlo shell model (MCSM) to study the E1 excitation in order to to treat larger model space. By using the MCSM we discuss some results about light nuclei. We discuss a microscopic description of E1 excitations based on shell-model calculations. We performed large-scale shell-model calculations for Ca isotopes with Lanczos-strength-function method and sd - pf - sdg model space allowing up to 3 ℏω excitation and obtained their photoabsorption cross sections. It gives a very good description of giant dipole and low-lying pygmy resonances rather independently of smoothing parameter. We also present the feasiblity of the Monte Carlo shell model (MCSM) to study the E1 excitation in order to to treat larger model space. By using the MCSM we discuss some results about light nuclei. This study is supported by HPCI strategic program field 5 and KAKENHI Grand 25870168.

  14. Dipole strength distributions in the stable Ba isotopes 134 138 Ba : A study in the mass region of a nuclear shape transition

    NASA Astrophysics Data System (ADS)

    Scheck, M.; von Garrel, H.; Tsoneva, N.; Belic, D.; von Brentano, P.; Fransen, C.; Gade, A.; Jolie, J.; Kneissl, U.; Kohstall, C.; Linnemann, A.; Nord, A.; Pietralla, N.; Pitz, H. H.; Stedile, F.; Stoyanov, C.; Werner, V.

    2004-10-01

    The low-lying dipole strength distributions in the odd-mass isotopes 135,137 Ba were studied in nuclear resonance fluorescence experiments (NRF) performed at the Stuttgart Dynamitron facility using bremsstrahlung beams with end point energies of 4.1, 3.1, and 2.5 MeV . Numerous excited states, most of them unknown so far, were observed in the excitation energy range up to 4 MeV . Detailed spectroscopic information has been obtained on excitation energies, decay widths, decay branching ratios, and transition probabilities. The results for 137Ba are compared with calculations in the framework of the Quasiparticle-Phonon Model. The new data for 135,137 Ba complete the systematics of low-lying dipole excitations as observed for the even Ba isotopes 134,136,138 Ba in previous NRF experiments in Stuttgart. The complete systematics within the Ba isotopic chain, exhibiting a nuclear shape transition, is discussed with respect to E1 two-phonon excitations, M1 scissors mode excitations, and in regard to the new critical point symmetries.

  15. Electric Monopole Transition Strengths in 62Ni

    NASA Astrophysics Data System (ADS)

    Evitts, L. J.; Garnsworthy, A. B.; Kibédi, T.; Moukaddam, M.; Alshahrani, B.; Eriksen, T. K.; Holt, J. D.; Hota, S. S.; Lane, G. J.; Lee, B. Q.; McCormick, B. P.; Palalani, N.; Reed, M. W.; Stroberg, S. R.; Stuchbery, A. E.

    2016-09-01

    Excited states in 62Ni were populated with a (p, p') reaction using the 14UD Pelletron accelerator at the Australian National University. Electric monopole transition strengths, ρ2(E0), were measured through simultaneous detection of the internal conversion electrons and γ rays emitted from the de-excitation of populated states, using the Super-e spectrometer coupled with a germanium detector. The strength of the 02+ to 01+ transition has been measured to be 77-34+23 × 10-3 and agrees with previously reported values. Upper limits have been placed on the 03+ to 01+ and 03+ to 02+ transitions. The measured ρ2(E0) value of the 22+ to 21+ transition in 62Ni has been measured for the first time and found to be one of the largest ρ2(E0) values measured to date in nuclei heavier than Ca. The low-lying states of 62Ni have previously been classified as one- and two-phonon vibrational states based on level energies. The measured electric quadrupole transition strengths are consistent with this interpretation. However as electric monopole transitions are forbidden between states which differ by one phonon number, the simple harmonic quadrupole vibrational picture is not suffcient to explain the large ρ2(E0) value for the 22+ to 21+ transition.

  16. 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

  17. 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)

  18. Monopole strength as a probe of nuclear shape mixing

    SciTech Connect

    Meyer, R.A.

    1987-08-17

    The monopole strength, MS, within a single set of nuclear shape excitations is compared with the MS between different shapes. After misconceptions are pointed out concerning the spin dependence of B(E2) values, MS properties are juxtaposed with gamma-ray and beta-decay properties of /sup 70/Se, /sup 96/Zr, /sup 102/Pd, and the N = 60 isotones to illustrate the utility of combined investigations and evidence is given for the observation of a two-phonon octupole multiplet. Finally, consideration is given to the dominance of the /sup 3/S/sub 1/ force in producing deformation in the N > 50 1g nuclei. 23 refs., 4 figs.

  19. 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...

  20. 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

  1. Systematics of photon strength functions

    NASA Astrophysics Data System (ADS)

    Firestone, Richard

    2015-10-01

    The photon strength of high energy E1 transitions is well described by Brink-Axel theory based on the contribution of the Giant Dipole Resonance. No adequate theory is available for M1 and E2 transitions which do not generally compete strongly with high energy E1 transitions. Measurements with the 57Fe(3He,3He') reaction at the Oslo cyclotron have revealed that the photon strength below 2 MeV greatly exceeds BA predictions. Similar results have been found for numerous other nuclides. In this paper I will discuss my analysis of the 56Fe(n,γ)57Fe reaction which we investigated with both cold neutrons from the Budapest Reactor and thermal neutrons from the Rez Reactor (Prague). A >99% complete 57Fe capture γ-ray decay scheme containing 449 γ-rays deexciting 100 levels has been constructed on the basis of γ-ray singles and γγ -coincidence data. The photon strengths for 90 primary γ-rays with energies ranging from 92-7646 keV were calculated and compared with the predictions of Brink-Axel (BA) theory. Excellent agreement has been attained for the high energy transitions while the strength below 2 MeV exceeds BA predictions confirming the earlier Oslo (3He,3He' γ) results. Photon strengths for another 95 secondary M1, E1, and E2 γ-rays were also determined to also exceed BA predictions for transitions below 4 MeV. The dependence of photon strength on level energy and the statistical distribution of photon strengths will also be discussed in this talk.

  2. Strength nutrition.

    PubMed

    Volek, Jeff S

    2003-08-01

    Muscle strength is determined by muscle size and factors related to neural recruitment. Resistance training is a potent stimulus for increasing muscle size and strength. These increases are, to a large extent, influenced and mediated by changes in hormones that regulate important events during the recovery process following exercise. Provision of nutrients in the appropriate amounts and at the appropriate times is necessary to optimize the recovery process. This review discusses the results of research that has examined the potential for nutrition and dietary supplements to impact the acute response to resistance exercise and chronic adaptations to resistance training. To date, the most promising strategies to augment gains in muscle size and strength appear to be consumption of protein-carbohydrate calories before and after resistance exercise, and creatine supplementation.

  3. Activation of NF-κB by Human Papillomavirus 16 E1 Limits E1-Dependent Viral Replication through Degradation of E1

    PubMed Central

    Nakahara, Tomomi; Tanaka, Katsuyuki; Ohno, Shin-ichi; Egawa, Nagayasu; Yugawa, Takashi

    2015-01-01

    ABSTRACT NF-κB is a family of transcription factors that regulate gene expression involved in many processes, such as the inflammatory response and cancer progression. Little is known about associations of NF-κB with the human papillomavirus (HPV) life cycle. We have developed a tissue culture system to conditionally induce E1-dependent replication of the human papillomavirus 16 (HPV16) genome in human cervical keratinocytes and found that expression of HPV16 E1, a viral helicase, results in reduction of IκBα and subsequent activation of NF-κB in a manner dependent on helicase activity. Exogenous expression of a degradation-resistant mutant of IκBα, which inhibits the activation of NF-κB, enhanced E1-dependent replication of the viral genome. Wortmannin, a broad inhibitor of phosphoinositide 3-kinases (PI3Ks), and, to a lesser extent, VE-822, an ATR kinase inhibitor, but not KU55933, an ATM kinase inhibitor, suppressed the activation of NF-κB and augmented E1-dependent replication of the HPV16 genome. Interestingly, the enhancement of E1-dependent replication of the viral genome was associated with increased stability of E1 in the presence of wortmannin as well as the IκBα mutant. Collectively, we propose that expression of E1 induces NF-κB activation at least in part through the ATR-dependent DNA damage response and that NF-κB in turn limits E1-dependent replication of HPV16 through degradation of E1, so that E1 and NF-κB may constitute a negative feedback loop. IMPORTANCE A major risk factor in human papillomavirus (HPV)-associated cancers is persistent infection with high-risk HPVs. To eradicate viruses from infected tissue, it is important to understand molecular mechanisms underlying the establishment and maintenance of persistent infection. In this study, we obtained evidence that human papillomavirus 16 (HPV16) E1, a viral DNA helicase essential for amplification of the viral genomes, induces NF-κB activation and that this limits E1-dependent

  4. E1, E2 and M1 transition parameters for some levels over ionization limit of Ne III

    NASA Astrophysics Data System (ADS)

    Eser, Selda; Özdemir, Leyla

    2016-07-01

    We have reported the level energies and radiative transition ( E1 , E2 and M1 parameters, such as wavelengths, transition rates, oscillator strengths and line strengths for some levels over the ionization limit of Ne III (oxygen-like). The calculations have been performed using the general-purpose relativistic atomic structure package (GRASP) based on the fully relativistic multiconfiguration Dirac-Fock (MCDF) method. The results obtained have been compared with the available theoretical and experimental values in the literature.

  5. 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.

  6. 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.

  7. 26 CFR 1.665(e)-1 - Preceding taxable year.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 8 2012-04-01 2012-04-01 false Preceding taxable year. 1.665(e)-1 Section 1.665... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning Before January 1, 1969 § 1.665(e)-1 Preceding taxable year. (a) Definition. For purposes...

  8. 26 CFR 1.665(e)-1 - Preceding taxable year.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 8 2011-04-01 2011-04-01 false Preceding taxable year. 1.665(e)-1 Section 1.665... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning Before January 1, 1969 § 1.665(e)-1 Preceding taxable year. (a) Definition. For purposes...

  9. 26 CFR 1.665(e)-1 - Preceding taxable year.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 8 2014-04-01 2014-04-01 false Preceding taxable year. 1.665(e)-1 Section 1.665... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning Before January 1, 1969 § 1.665(e)-1 Preceding taxable year. (a) Definition. For purposes...

  10. 26 CFR 1.514(e)-1 - Allocation rules.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 7 2010-04-01 2010-04-01 true Allocation rules. 1.514(e)-1 Section 1.514(e)-1... Allocation rules. Where only a portion of property is debt-financed property, proper allocation of the basis...)(iii) of § 1.514(b)-1 for illustrations of proper allocation....

  11. 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...

  12. Effects of Mutations in the Rubella Virus E1 Glycoprotein on E1-E2 Interaction and Membrane Fusion Activity

    PubMed Central

    Yang, Decheng; Hwang, Dorothy; Qiu, Zhiyong; Gillam, Shirley

    1998-01-01

    Rubella virus (RV) virions contain two glycosylated membrane proteins, E1 and E2, that exist as a heterodimer and form the viral spike complexes on the virion surface. Formation of an E1-E2 heterodimer is required for transport of E1 out of the endoplasmic reticulum lumen to the Golgi apparatus and plasma membrane. To investigate the nature of the E1-E2 interaction, we have introduced mutations in the internal hydrophobic region (residues 81 to 109) of E1. Substitution of serine at Cys82 (mutant C82S) or deletion of this hydrophobic domain (mutant dt) of E1 resulted in a disruption of the E1 conformation that ultimately affected E1-E2 heterodimer formation and cell surface expression of both E1 and E2. Substitution of either aspartic acid at Gly93 (G93D) or glycine at Pro104 (P104G) was found to impair neither E1-E2 heterodimer formation nor the transport of E1 and E2 to the cell surface. Fusion of RV-infected cells is induced by a brief treatment at a pH below 6.0. To test whether this internal hydrophobic domain is involved in the membrane fusion activity of RV, transformed BHK cell lines expressing either wild-type or mutant spike proteins were exposed to an acidic pH and polykaryon formation was measured. No fusion activity was observed in the C82S, dt, and G93D mutants; however, the wild type and the P104G mutant exhibited fusogenic activities, with greater than 60% and 20 to 40% of the cells being fused, respectively, at pH 4.8. These results suggest that it is likely that the region of E1 between amino acids 81 and 109 is involved in the membrane fusion activity of RV and that it may be important for the interaction of that protein with E2 to form the E1-E2 heterodimer. PMID:9765418

  13. CYP2E1 hydroxylation of aniline involves negative cooperativity.

    PubMed

    Hartman, Jessica H; Knott, Katie; Miller, Grover P

    2014-02-01

    CYP2E1 plays a role in the metabolic activation and elimination of aniline, yet there are conflicting reports on its mechanism of action, and hence relevance, in aniline metabolism. Based on our work with similar compounds, we hypothesized that aniline binds two CYP2E1 sites during metabolism resulting in cooperative reaction kinetics and tested this hypothesis through rigorous in vitro studies. The kinetic profile for recombinant CYP2E1 demonstrated significant negative cooperativity based on a fit of data to the Hill equation (n=0.56). Mechanistically, the data were best explained through a two-binding site cooperative model in which aniline binds with high affinity (K(s)=30 μM) followed by a second weaker binding event (K(ss)=1100 uM) resulting in a threefold increase in the oxidation rate. Binding sites for aniline were confirmed by inhibition studies with 4-methylpyrazole. Inhibitor phenotyping experiments with human liver microsomes validated the central role for CYP2E1 in aniline hydroxylation and indicated minor roles for CYP2A6 and CYP2C9. Importantly, inhibition of minor metabolic pathways resulted in a kinetic profile for microsomal CYP2E1 that replicated the preferred mechanism and parameters observed with the recombinant enzyme. Scaled modeling of in vitro CYP2E1 metabolism of aniline to in vivo clearance, especially at low aniline levels, led to significant deviations from the traditional model based on non-cooperative, Michaelis-Menten kinetics. These findings provide a critical mechanistic perspective on the potential importance of CYP2E1 in the metabolic activation and elimination of aniline as well as the first experimental evidence of a negatively cooperative metabolic reaction catalyzed by CYP2E1.

  14. E1a promotes c-Myc-dependent replicative stress

    PubMed Central

    Valero, María Llanos; Cimas, Francisco Jose; Arias, Laura; Melgar-Rojas, Pedro; García, Elena; Callejas-Valera, Juan Luis; García-Cano, Jesús; Serrano-Oviedo, Leticia; Ángel de la Cruz-Morcillo, Miguel; Sánchez-Pérez, Isabel; Sánchez-Prieto, Ricardo

    2014-01-01

    The E1a gene from adenovirus is known to be a potent inducer of chemo/radiosensitivity in a wide range of tumors. However, the molecular bases of its radiosensitizer properties are still poorly understood. In an attempt to study this effect, U87MG cells, derived from a radio-resistant tumor as glioblastoma, where infected with lentivirus carrying E1a gene developing an acute sensitivity to ionizing radiation. The induction of radiosensitivity correlated with a marked G2/M phase accumulation and a potent apoptotic response. Our findings demonstrate that c-Myc plays a pivotal role in E1a-associated radiosensitivity through the induction of a replicative stress situation, as our data support by genetic approaches, based in interference and overexpression in U87MG cells. In fact, we present evidence showing that Chk1 is a novel transcriptional target of E1a gene through the effect exerted by this adenoviral protein onto c-Myc. Moreover, c-Myc upregulation also explains the marked phosphorylation of H2AX associated to E1a expression in the absence of DNA damage. Indeed, all these observations were applicable to other experimental models, such as T98G, LN-405 and A172, rendering the same pattern in terms of radiosensitivity, cell cycle distribution, upregulation of Chk1, c-Myc, and phosphorylation pattern of H2AX. In summary, our data propose a novel mechanism to explain how E1a mediates radiosensitivity through the signaling axis E1a→c-Myc→ replicative stress situation. This novel mechanism of E1a-mediated radiosensitivity could be the key to open new possibilities in the current therapy of glioblastoma. PMID:24196438

  15. Resolvin E1 Reverses Experimental Periodontitis and Dysbiosis.

    PubMed

    Lee, Chun-Teh; Teles, Ricardo; Kantarci, Alpdogan; Chen, Tsute; McCafferty, Jon; Starr, Jacqueline R; Brito, Luciana Carla Neves; Paster, Bruce J; Van Dyke, Thomas E

    2016-10-01

    Periodontitis is a biofilm-induced inflammatory disease characterized by dysbiosis of the commensal periodontal microbiota. It is unclear how natural regulation of inflammation affects the periodontal biofilm. Promoters of active resolution of inflammation, including resolvin E1 (RvE1), effectively treat inflammatory periodontitis in animal models. The goals of this study were 1) to compare periodontal tissue gene expression in different clinical conditions, 2) to determine the impact of local inflammation on the composition of subgingival bacteria, and 3) to understand how inflammation impacts these changes. Two clinically relevant experiments were performed in rats: prevention and treatment of ligature-induced periodontitis with RvE1 topical treatment. The gingival transcriptome was evaluated by RNA sequencing of mRNA. The composition of the subgingival microbiota was characterized by 16S rDNA sequencing. Periodontitis was assessed by bone morphometric measurements and histomorphometry of block sections. H&E and tartrate-resistant acid phosphatase staining were used to characterize and quantify inflammatory changes. RvE1 treatment prevented bone loss in ligature-induced periodontitis. Osteoclast density and inflammatory cell infiltration in the RvE1 groups were lower than those in the placebo group. RvE1 treatment reduced expression of inflammation-related genes, returning the expression profile to one more similar to health. Treatment of established periodontitis with RvE1 reversed bone loss, reversed inflammatory gene expression, and reduced osteoclast density. Assessment of the rat subgingival microbiota after RvE1 treatment revealed marked changes in both prevention and treatment experiments. The data suggest that modulation of local inflammation has a major role in shaping the composition of the subgingival microbiota.

  16. DprE1, a new taxonomic marker in mycobacteria.

    PubMed

    Incandela, Maria Loreto; Perrin, Elena; Fondi, Marco; de Jesus Lopes Ribeiro, Ana Luisa; Mori, Giorgia; Moiana, Alessia; Gramegna, Maurizio; Fani, Renato; Riccardi, Giovanna; Pasca, Maria Rosalia

    2013-11-01

    Among the species of the Mycobacterium genus, more than 50 have been recognized as human pathogens. In spite of the different diseases caused by mycobacteria, the interspecies genetic similarity ranges from 94% to 100%, and for some species, this value is higher than in other bacteria. Consequently, it is important to understand the relationships existing among mycobacterial species. In this context, the possibility to use Mycobacterium tuberculosis dprE1 gene as new phylogenetic/taxonomic marker has been explored. The dprE1 gene codes for the target of benzothiazinones, belonging to a very promising class of antitubercular drugs. Mutations in cysteine 387 of DprE1 are responsible for benzothiazinone resistance. The DprE1 tree, obtained with 73 amino acid sequences of mycobacterial species, revealed that concerning the benzothiazinone sensitivity/resistance, it is possible to discriminate two clusters. To validate it, a concatamer obtained from the amino acid sequences of nine mycobacterial housekeeping genes was performed. The concatamer revealed that there is no separation between the benzothiazinone-susceptible and benzothiazinone-resistant species; consequently, this parameter is not linked to the phylogeny. DprE1 tree might represent a good taxonomic marker for the assignment of a mycobacterial isolate to a species. Moreover, the concatamer represents a good reference phylogeny for the Mycobacterium genus. PMID:24024613

  17. Impotence evaluated by the use of prostaglandin E1

    SciTech Connect

    Hwang, T.I.; Yang, C.R.; Wang, S.J.; Chang, C.L.; Tzai, T.S.; Chang, C.H.; Wu, H.C.

    1989-06-01

    We screened 80 patients at our hospital for the differential diagnosis of impotence using intracavernous injection of prostaglandin E1 (20 micrograms). The rate of positive response was 78.8 per cent (63 patients). Neither systemic reactions nor priapism occurred. However, a considerable incidence (23.8 per cent, 19 of 80 patients) of tolerable injection pain was encountered. The 133-xenon penile washout study was conducted routinely in impotent men for hemodynamic evaluation of penile vascularity. In 80 patients a positive correlation between the response of intracavernous prostaglandin E1 injection and the result of the washout study was found (r equals 0.381, p less than 0.0002). We selected 14 subjects randomly to receive additional intravenous infusions of prostaglandin E1 (6 ampules, 120 micrograms total) for 3 days, after which another 133-xenon washout study was done. The washout studies before and after intravenous prostaglandin E1 infusion were compared, and 10 patients (71.4 per cent) appeared to obtain improvement in half-time clearance and penile blood flow. However, only 3 patients noticed improvement subjectively. We suggest that prostaglandin E1 could be a desirable alternative for the diagnosis and treatment of impotence.

  18. 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...

  19. 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

  20. 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…

  1. A simple method for the simultaneous detection of E1A and E1B in adenovirus stocks.

    PubMed

    Suzuki, Erika; Murata, Takehide; Watanabe, Sanae; Kujime, Yukari; Hirose, Megumi; Pan, Jianzhi; Yamazaki, Takahito; Ugai, Hideyo; Yokoyama, Kazunari K

    2004-01-01

    Recombinant adenoviral vectors have been developed for use as therapeutic agents and for the introduction of exogenous genes into living cells. However, the occurrence of replication-competent adenoviruses (RCA) in adenovirus stocks produced in 293 cells remains a major problem in terms of the safe use of such vectors. To overcome the problems associated with the occurrence of RCA, we have established a simple method for the simultaneous detection of amplified E1A and E1B from RCA that might contaminate adenoviral stocks. The products amplified by polymerase chain reaction (PCR) were fractionated by regular electrophoresis on agarose gels and visualized by staining with ethidium bromide. This method is rapid and inexpensive for detection of RCA in the preparation of adenoviruses. PMID:14654922

  2. A simple method for the simultaneous detection of E1A and E1B in adenovirus stocks.

    PubMed

    Suzuki, Erika; Murata, Takehide; Watanabe, Sanae; Kujime, Yukari; Hirose, Megumi; Pan, Jianzhi; Yamazaki, Takahito; Ugai, Hideyo; Yokoyama, Kazunari K

    2004-01-01

    Recombinant adenoviral vectors have been developed for use as therapeutic agents and for the introduction of exogenous genes into living cells. However, the occurrence of replication-competent adenoviruses (RCA) in adenovirus stocks produced in 293 cells remains a major problem in terms of the safe use of such vectors. To overcome the problems associated with the occurrence of RCA, we have established a simple method for the simultaneous detection of amplified E1A and E1B from RCA that might contaminate adenoviral stocks. The products amplified by polymerase chain reaction (PCR) were fractionated by regular electrophoresis on agarose gels and visualized by staining with ethidium bromide. This method is rapid and inexpensive for detection of RCA in the preparation of adenoviruses.

  3. 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

  4. Identification of strong E1 and M1 groundstate transitions in deformed rare earth nuclei

    NASA Astrophysics Data System (ADS)

    Friedrichs, H.; Lindenstruth, S.; Schlitt, B.; Wesselborg, C.; Bauske, I.; Heil, R. D.; Kneissl, U.; Margral, J.; Pitz, H. H.; Häger, D.; Müller, G.; Schumacher, M.; von Brentano, P.; Herzberg, R. D.; Zilges, A.

    1993-03-01

    Systematic nuclear resonance fluorescence (NRF) experiments have been performed at the bremsstrahlung facility of the Stuttgart Dynamitron to investigate the distribution of magnetic and electric dipole strengths in deformed nuclei. Precise excitation energies, transition strengths, spins and decay branching ratios were deduced for numerous low lying dipole excitations in deformed rare earth nuclei. Measurements of the linear polarization of resonantly scattered photons using simultaneously two Compton polarimeters enabled model independent parity assignments. For the first time positive parities could be established for groups of states in the neighbouring deformed nuclei 150Nd, 160Gd, 162Dy. Most of these states are concentrated near 3 MeV and should be attributed to orbital M1 excitations ("Scissors Mode"). The deformation dependence of the orbital M1 strength has been studied in the Nd isotopic chain. Completing previous polarization measurements on 142,150Nd the transitional nucleus 146Nd has been investigated. The surprising novel result of the present systematic studies, however, was the first observation of enhanced electric dipole excitations in the same deformed nuclei at excitation energies of 2.414, 2.471, and 2.520 MeV, respectively. The transition energies and the enhanced B(E1)↑ strengths of 3-5·10-3e2fm2 support the interpretation in terms of the predicted new type of collective electric dipole excitations in deformed nuclei due to reflection asymmetric shapes like octupole deformations and/or cluster configurations. Furthermore, all three states systematically exhibit decay branching ratios Rexp = B(1- → 21+)/B(1- → 01+), which hint at K-mixing. First results for the neutron-odd, deformed nucleus 163Dy are presented.

  5. 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.

  6. Search for two-phonon octupole excitations in 146Gd

    NASA Astrophysics Data System (ADS)

    Orce, J. N.; Kumar Raju, M.; Khumalo, N. A.; Dinoko, T. S.; Jones, P.; Bark, R. A.; Lawrie, E. A.; Majola, S. N. T.; Robledo, L. M.; Rubio, B.; Wiedeking, M.; Easton, J.; Khaleel, E. A.; Kheswa, B. V.; Kheswa, N.; Herbert, M. S.; Lawrie, J. J.; Masiteng, P. L.; Nchodu, M. R.; Ndayishimye, J.; Negi, D.; Noncolela, S. P.; Ntshangase, S. S.; Papka, P.; Roux, D. G.; Shirinda, O.; Sithole, P. S.; Yates, S. W.

    2016-06-01

    The low-spin structure of the nearly spherical nucleus 146Gd was studied using the 144Sm(4He, 2n) fusion-evaporation reaction. High-statistics γ - γ coincidence measurements were performed at iThemba LABS with 7× 109 γ- γ coincidence events recorded. Gated γ-ray energy spectra show evidence for the 6+2 → 3-1 → 0+1 cascade of E3 transitions in agreement with recent findings by Caballero and co-workers, but with a smaller branching ratio of I_{γ} = 4.7(10) for the 6+2 → 3-1 1905.1 keV γ ray. Although these findings may support octupole vibrations in spherical nuclei, sophisticated beyond mean-field calculations including angular-momentum projection are required to interpret in an appropriate way the available data due to the failure of the rotational model assumptions in this nucleus.

  7. Metabolism of prostaglandin E1 in dog kidneys

    PubMed Central

    Nakano, J.

    1970-01-01

    1. The biotransformation of prostaglandin E1 (PGE1) was studied in the isolated, perfused dog kidneys. 2. An average 43% of PGE1 was converted into the less polar metabolite I by a single passage through the kidney. As the re-circulation of the perfusate continued, PGE1 was converted not only into metabolite I but also the least polar metabolite II. The velocity of the conversion of PGE1 into metabolite I was significantly greater than that into metabolite II. Usually, six passages elapsed before maximum degradation of PGE1 occurred. 3. Further separation with silicic acid column chromatography and gas-liquid chromatography showed that metabolite II consists of two individual metabolites, metabolite IIa and metabolite IIb. 4. The present study indicates that the kidney biotransforms PGE1 rather rapidly into three metabolites which are less polar than PGE1. PMID:5492900

  8. Tumorigenicity and adenovirus-transformed cells: Collagen interaction and cell surface laminin are controlled by the serotype origin of the E1A and E1B genes

    SciTech Connect

    Bober, F.J.; Birk, D.E.; Raska, K. Jr. ); Shenk, T. )

    1988-02-01

    A library of cells transformed with recombinant adenoviruses was used to study tumorigenicity and interaction with extracellular matrix. Cells expressing the complete E1 region of highly oncogenic adenovirus type 12 (Ad12) are tumorigenic, adhere preferentially to type IV collagen, and express cell surface laminin. Weakly tumorigenic cells, which express the E1A oncogene of Ad12 and the E1B genes of Ad5, also attach preferentially to type IV collagen but do not contain laminin on their surface. Cells which express the E1A oncogene of Ad5 and the E1B genes of Ad12 are nontumorigenic and do not preferentially attach to type IV versus type I collagen but have laminin on their surface. There is no significant difference in the amounts of laminin secreted into the culture medium among cells expressing the E1B genes of Ad5 or Ad12. In vitro assays show that cells which express the E1B genes of Ad12, irrespective of the origin of the E1A genes, can bind three times more exogenously added {sup 125}I-laminin than cells expressing the E1B genes of nononcogenic Ad5. The interaction of adenovirus-transformed cells with collagen is controlled by the serotype origin of the E1A oncogene, whereas cell surface laminin is controlled by the serotype origin of the E1B genes.

  9. E1-forbidden transition rates in ions of astrophysical interest

    NASA Astrophysics Data System (ADS)

    Träbert, E.

    2014-11-01

    Transition rates in atomic systems may appear to be of little importance in steady-state plasmas that are observed at great distances from Earth. However, some of the transition rates compete with collision rates, and in these cases certain line intensity ratios are affected and can serve as remote indicators of density. In the low-density environments of stellar coronae and planetary nebulae, the transition rates of interest are mostly spin-forbidden E1 decays, higher-multipole order transitions (M1, E2, M2, M3), and hyperfine-induced transitions. On Earth, measurements of the long upper level lifetimes of these atomic systems require the use of ion traps. A fair number of test cases with lifetimes in the range from nanoseconds to many seconds have been treated successfully, and the evolution of calculations along with the experimental progress is notable. A new generation of cold ion traps is expected to extend the atomic lifetime measurements on multiply charged ions into the range of many minutes.

  10. 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)

  11. 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

  12. What Is a Strength?

    ERIC Educational Resources Information Center

    Wolin, Sybil

    2003-01-01

    As the strength-based perspective gains recognition, it is important to describe what constitutes strengths and to develop a specific vocabulary to name them. This article draws on resilience research to help identify specific competencies and areas of strengths in youth. (Contains 1 table.)

  13. 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…

  14. Enhancement of octupole strength in near spherical nuclei

    NASA Astrophysics Data System (ADS)

    Robledo, L. M.

    2016-09-01

    The validity of the rotational formula used to compute E1 and E3 transition strengths in even-even nuclei is analyzed within the Generator Coordinate Method framework based on mean field wave functions. It turns out that those nuclei with spherical or near spherical shapes the E1 and E3 strengths computed with this formula are strongly underestimated and a sound evaluation of them requires angular-momentum projected wave functions. Results for several isotopic chains with proton number equal to or near magic numbers are analyzed and compared with experimental data. The use of angular-momentum projected wave functions greatly improves the agreement with the scarce experimental data.

  15. 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.

  16. 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

  17. Conformational anti-cytochrome P4502E1 (CYP2E1) auto-antibodies contribute to necro-inflammatory injury in chronic hepatitis C.

    PubMed

    Sutti, S; Vidali, M; Mombello, C; Sartori, M; Albano, E

    2010-10-01

    Circulating auto-antibodies against cytochrome P4502E1 (CYP2E1) have been observed in a significant fraction of patients with chronic hepatitis C (CHC). This study investigated the clinical significance of these auto-antibodies in relation to their antigen specificity. The presence of anti-CYP2E1 IgG was investigated in 137 consecutive patients with biopsy-proven CHC. Anti-CYP2E1 IgG above control threshold levels was detected in 52 (38%) subjects. By combined immunoprecipitation and western blotting, we observed that among anti-CYP2E1 IgG-positive sera, 23 (44%) were unreactive towards denaturated CYP2E1, indicating a prevalent recognition of conformational CYP2E1 antigens. Conformational anti-CYP2E1 auto-antibodies were unrelated to circulating gamma-globulins, alcohol intake or infection by specific HCV genotypes. The presence of anti-CYP2E1 auto-antibodies was associated with an 11-fold (OR 10.9 95%CI 1.4-86.6 P = 0.008) increased prevalence of necro-inflammatory grading ≥ 4 (Ishack's criteria) and 4-fold (OR 4.0; 95%CI 1.3-11-7: P = 0.014) increased prevalence of fibrosis staging ≥ 2, respectively. Multivariate analysis confirmed conformational anti-CYP2E1 IgG (P = 0.005) and age (P = 0.033) as independent predictors of necro-inflammatory grading ≥ 4. The development of anti-CYP2E1 auto-antibodies targeting conformational CYP2E1 epitopes is associated with more severe liver damage in CHC.

  18. The Human Adenovirus Type 5 E4orf6/E1B55K E3 Ubiquitin Ligase Complex Enhances E1A Functional Activity.

    PubMed

    Dallaire, Frédéric; Schreiner, Sabrina; Blair, G Eric; Dobner, Thomas; Branton, Philip E; Blanchette, Paola

    2016-01-01

    Human adenovirus (Ad) E1A proteins have long been known as the central regulators of virus infection as well as the major source of adenovirus oncogenic potential. Not only do they activate expression of other early viral genes, they make viral replication possible in terminally differentiated cells, at least in part, by binding to the retinoblastoma (Rb) tumor suppressor family of proteins to activate E2F transcription factors and thus viral and cellular DNA synthesis. We demonstrate in an accompanying article (F. Dallaire et al., mSphere 1:00014-15, 2016) that the human adenovirus E3 ubiquitin ligase complex formed by the E4orf6 and E1B55K proteins is able to mimic E1A activation of E2F transactivation factors. Acting alone in the absence of E1A, the Ad5 E4orf6 protein in complex with E1B55K was shown to bind E2F, disrupt E2F/Rb complexes, and induce hyperphosphorylation of Rb, leading to induction of viral and cellular DNA synthesis, as well as stimulation of early and late viral gene expression and production of viral progeny. While these activities were significantly lower than those exhibited by E1A, we report here that this ligase complex appeared to enhance E1A activity in two ways. First, the E4orf6/E1B55K complex was shown to stabilize E1A proteins, leading to higher levels in infected cells. Second, the complex was demonstrated to enhance the activation of E2F by E1A products. These findings indicated a new role of the E4orf6/E1B55K ligase complex in promoting adenovirus replication. IMPORTANCE Following our demonstration that adenovirus E3 ubiquitin ligase formed by the viral E4orf6 and E1B55K proteins is able to mimic the activation of E2F by E1A, we conducted a series of studies to determine if this complex might also promote the ability of E1A to do so. We found that the complex both significantly stabilizes E1A proteins and also enhances their ability to activate E2F. This finding is of significance because it represents an entirely new function for

  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. Metabolic inactivation of resolvin E1 and stabilization of its anti-inflammatory actions.

    PubMed

    Arita, Makoto; Oh, Sungwhan F; Chonan, Tomomichi; Hong, Song; Elangovan, Siva; Sun, Yee-Ping; Uddin, Jasim; Petasis, Nicos A; Serhan, Charles N

    2006-08-11

    The resolvins (Rv) are lipid mediators derived from omega-3 polyunsaturated fatty acids that act within a local inflammatory milieu to stop leukocyte recruitment and promote resolution. Resolvin E1 (RvE1; (5S,12R,18R)-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid) is an oxygenase product derived from omega-3 eicosapentaenoic acid that displays potent anti-inflammation/pro-resolution actions in vivo. Here, we determined whether oxidoreductase enzymes catalyze the conversion of RvE1 and assessed the biological activity of the RvE1 metabolite. With NAD+ as a cofactor, recombinant 15-hydroxyprostaglandin dehydrogenase acted as an 18-hydroxyl dehydrogenase to form 18-oxo-RvE1. In the murine lung, dehydrogenation of the hydroxyl group at carbon 18 position to form 18-oxo-RvE1 represented the major initial metabolic route for RvE1. At a concentration where RvE1 potently reduced polymorphonuclear leukocyte (PMN) recruitment in zymosan-induced peritonitis, 18-oxo-RvE1 was devoid of activity. In human neutrophils, carbon 20 hydroxylation of RvE1 was the main route of conversion. An RvE1 analog, i.e. 19-(p-fluorophenoxy)-RvE1, was synthesized that resisted rapid metabolic inactivation and proved to retain biological activity reducing PMN infiltration and pro-inflammatory cytokine/chemokine production in vivo. These results established the structure of a novel RvE1 initial metabolite, indicating that conversion of RvE1 to the oxo product represents a mode of RvE1 inactivation. Moreover, the designed RvE1 analog, which resisted further metabolism/inactivation, could be a useful tool to evaluate the actions of RvE1 in complex disease models.

  1. Metabolic inactivation of resolvin E1 and stabilization of its anti-inflammatory actions.

    PubMed

    Arita, Makoto; Oh, Sungwhan F; Chonan, Tomomichi; Hong, Song; Elangovan, Siva; Sun, Yee-Ping; Uddin, Jasim; Petasis, Nicos A; Serhan, Charles N

    2006-08-11

    The resolvins (Rv) are lipid mediators derived from omega-3 polyunsaturated fatty acids that act within a local inflammatory milieu to stop leukocyte recruitment and promote resolution. Resolvin E1 (RvE1; (5S,12R,18R)-trihydroxy-6Z,8E,10E,14Z,16E-eicosapentaenoic acid) is an oxygenase product derived from omega-3 eicosapentaenoic acid that displays potent anti-inflammation/pro-resolution actions in vivo. Here, we determined whether oxidoreductase enzymes catalyze the conversion of RvE1 and assessed the biological activity of the RvE1 metabolite. With NAD+ as a cofactor, recombinant 15-hydroxyprostaglandin dehydrogenase acted as an 18-hydroxyl dehydrogenase to form 18-oxo-RvE1. In the murine lung, dehydrogenation of the hydroxyl group at carbon 18 position to form 18-oxo-RvE1 represented the major initial metabolic route for RvE1. At a concentration where RvE1 potently reduced polymorphonuclear leukocyte (PMN) recruitment in zymosan-induced peritonitis, 18-oxo-RvE1 was devoid of activity. In human neutrophils, carbon 20 hydroxylation of RvE1 was the main route of conversion. An RvE1 analog, i.e. 19-(p-fluorophenoxy)-RvE1, was synthesized that resisted rapid metabolic inactivation and proved to retain biological activity reducing PMN infiltration and pro-inflammatory cytokine/chemokine production in vivo. These results established the structure of a novel RvE1 initial metabolite, indicating that conversion of RvE1 to the oxo product represents a mode of RvE1 inactivation. Moreover, the designed RvE1 analog, which resisted further metabolism/inactivation, could be a useful tool to evaluate the actions of RvE1 in complex disease models. PMID:16757471

  2. Functional conservation and diversification of the soybean maturity gene E1 and its homologs in legumes

    PubMed Central

    Zhang, Xingzheng; Zhai, Hong; Wang, Yaying; Tian, Xiaojie; Zhang, Yupeng; Wu, Hongyan; Lü, Shixiang; Yang, Guang; Li, Yuqiu; Wang, Lu; Hu, Bo; Bu, Qingyun; Xia, Zhengjun

    2016-01-01

    Gene regulatory networks involved in flowering time and photoperiodic responses in legumes remain unknown. Although the major maturity gene E1 has been successfully deciphered in soybean, knowledge on the functional conservation of this gene is limited to a certain extent to E1 homologs in legumes. The ectopic expression of Phvul.009G204600 (PvE1L), an E1 homolog from common bean, delayed the onset of flowering in soybean. By contrast, the ectopic expression of Medtr2g058520 (MtE1L) from Medicago truncatula did not affect the flowering of soybean. Characterization of the late-flowering mte1l mutant indicated that MtE1L promoted flowering in Medicago truncatula. Moreover, all transgenic E1, PvE1L and MtE1L soybean lines exhibited phenotypic changes in terms of plant height. Transgenic E1 or PvE1L plants were taller than the wild-type, whereas transgenic MtE1L plants produced dwarf phenotype with few nodes and short internode. Thus, functional conservation and diversification of E1 family genes from legumes in the regulation of flowering and plant growth may be associated with lineage specification and genomic duplication. PMID:27405888

  3. Functional conservation and diversification of the soybean maturity gene E1 and its homologs in legumes.

    PubMed

    Zhang, Xingzheng; Zhai, Hong; Wang, Yaying; Tian, Xiaojie; Zhang, Yupeng; Wu, Hongyan; Lü, Shixiang; Yang, Guang; Li, Yuqiu; Wang, Lu; Hu, Bo; Bu, Qingyun; Xia, Zhengjun

    2016-01-01

    Gene regulatory networks involved in flowering time and photoperiodic responses in legumes remain unknown. Although the major maturity gene E1 has been successfully deciphered in soybean, knowledge on the functional conservation of this gene is limited to a certain extent to E1 homologs in legumes. The ectopic expression of Phvul.009G204600 (PvE1L), an E1 homolog from common bean, delayed the onset of flowering in soybean. By contrast, the ectopic expression of Medtr2g058520 (MtE1L) from Medicago truncatula did not affect the flowering of soybean. Characterization of the late-flowering mte1l mutant indicated that MtE1L promoted flowering in Medicago truncatula. Moreover, all transgenic E1, PvE1L and MtE1L soybean lines exhibited phenotypic changes in terms of plant height. Transgenic E1 or PvE1L plants were taller than the wild-type, whereas transgenic MtE1L plants produced dwarf phenotype with few nodes and short internode. Thus, functional conservation and diversification of E1 family genes from legumes in the regulation of flowering and plant growth may be associated with lineage specification and genomic duplication. PMID:27405888

  4. 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, Steven 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.

  5. 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.

  6. 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... by section 4216(e) and paragraph (c) of this section has application only if: (1) In the case...

  7. 26 CFR 1.401(e)-1 - Definitions relating to plans covering self-employed individuals.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...-employed individuals. 1.401(e)-1 Section 1.401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT... Plans, Etc. § 1.401(e)-1 Definitions relating to plans covering self-employed individuals. (a) “Keogh... is subject to the rules contained in §§ 1.401 (e)-2, (e)-5, and (j)-1 through (j)-5. Section...

  8. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... when prior law applied. 1.691(e)-1 Section 1.691(e)-1 Internal Revenue INTERNAL REVENUE SERVICE... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In general—(1) Application of prior law. Under section 44(d) of the Internal Revenue Code of 1939...

  9. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... when prior law applied. 1.691(e)-1 Section 1.691(e)-1 Internal Revenue INTERNAL REVENUE SERVICE... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In general—(1) Application of prior law. Under section 44(d) of the Internal Revenue Code of 1939...

  10. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... when prior law applied. 1.691(e)-1 Section 1.691(e)-1 Internal Revenue INTERNAL REVENUE SERVICE... Decedents § 1.691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In general—(1) Application of prior law. Under section 44(d) of the Internal Revenue Code of 1939...

  11. 26 CFR 31.3401(e)-1 - Number of withholding exemptions claimed.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... TAX AT SOURCE Collection of Income Tax at Source § 31.3401(e)-1 Number of withholding exemptions... 26 Internal Revenue 15 2011-04-01 2011-04-01 false Number of withholding exemptions claimed. 31.3401(e)-1 Section 31.3401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  12. 26 CFR 31.3401(e)-1 - Number of withholding exemptions claimed.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... TAX AT SOURCE Collection of Income Tax at Source § 31.3401(e)-1 Number of withholding exemptions... 26 Internal Revenue 15 2010-04-01 2010-04-01 false Number of withholding exemptions claimed. 31.3401(e)-1 Section 31.3401(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE...

  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. Facile synthesis of covalent probes to capture enzymatic intermediates during E1 enzyme catalysis.

    PubMed

    An, Heeseon; Statsyuk, Alexander V

    2016-02-11

    We report a facile synthetic strategy to prepare UBL-AMP electrophilic probes that form a covalent bond with the catalytic cysteine of cognate E1s, mimicking the tetrahedral intermediate of the E1-UBL-AMP complex. These probes enable the structural and biochemical study of both canonical- and non-canonical E1s.

  15. 26 CFR 48.6416(e)-1 - Refund to exporter or shipper.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... of Special Application to Retailers and Manufacturers Taxes § 48.6416(e)-1 Refund to exporter or... 26 Internal Revenue 16 2010-04-01 2010-04-01 true Refund to exporter or shipper. 48.6416(e)-1 Section 48.6416(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY...

  16. Strength Training and Your Child

    MedlinePlus

    ... Story" 5 Things to Know About Zika & Pregnancy Strength Training and Your Child KidsHealth > For Parents > Strength Training ... help prevent injuries and speed up recovery. About Strength Training Strength training is the practice of using free ...

  17. Building on Our Strengths.

    ERIC Educational Resources Information Center

    Hill, Robert

    1978-01-01

    Comments on the feeling that the American family is disintegrating, and that many criticisms traditionally made about Black families are now made about White families. Suggests that people need to stress family strengths. As an example, five major strengths of Black families are described: flexibility, work and achievement ethics, religiosity, and…

  18. Strengths of Remarried Families.

    ERIC Educational Resources Information Center

    Knaub, Patricia Kain; And Others

    1984-01-01

    Focuses on remarried families' (N=80) perceptions of family strengths, marital satisfaction, and adjustment to the remarried situation. Results indicated that although most would like to make some changes, scores on the measurements used were high. A supportive environment was the most important predictor of family strength and success. (JAC)

  19. Identification and characterization of an hnRNP E1 translational silencing motif

    PubMed Central

    Brown, Andrew S.; Mohanty, Bidyut K.; Howe, Philip H.

    2016-01-01

    Non-canonical transforming growth factor β (TGFβ) signaling through protein kinase B (Akt2) induces phosphorylation of heterogeneous nuclear ribonucleoprotein E1 (hnRNP E1) at serine-43 (p-hnRNP E1). This post-translational modification (PTM) of hnRNP E1 promotes its dissociation from a 3′ untranslated region (UTR) nucleic acid regulatory motif, driving epithelial to mesenchymal transition (EMT) and metastasis. We have identified an hnRNP E1 consensus-binding motif and genomically resolved a subset of genes in which it is contained. This study characterizes the binding kinetics of the consensus-binding motif and hnRNP E1, its various K-homology (KH) domains and p-hnRNP E1. Levels of p-hnRNP E1 are highly upregulated in metastatic cancer cells and low in normal epithelial tissue. We show a correlation between this PTM and levels of Akt2 and its activated form, phosphorylated serine-474 (p-Akt2). Using cellular progression models of metastasis, we observed a signature high level of Akt2, p-Akt2 and p-hnRNP E1 protein expression, coupled to a significantly reduced level of total hnRNP E1 in metastatic cells. Genes that are translationally silenced by hnRNP E1 and expressed by its dissociation are highly implicated in the progression of EMT and metastasis. This study provides insight into a non-canonical TGFβ signaling cascade that is responsible for inducing EMT by aberrant expression of hnRNP E1 silenced targets. The relevance of this system in metastatic progression is clearly shown in cellular models by the high abundance of p-hnRNP E1 and low levels of hnRNP E1. New insights provided by the resolution of this molecular mechanism provide targets for therapeutic intervention and give further insight into the role of the TGFβ microenvironment. PMID:27067543

  20. 17 CFR 274.127e-1 - Form N-27E-1, notice to periodic payment plan certificate holders of 18-month surrender rights...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    .... Editorial Note: For Federal Register citations affecting Form N-27E-1, see the List of CFR Sections Affected... EXCHANGE COMMISSION (CONTINUED) FORMS PRESCRIBED UNDER THE INVESTMENT COMPANY ACT OF 1940 Forms for...

  1. [Induction of rat hepatic CYP2E1 expression by arecoline in vivo].

    PubMed

    Huang, Xiang-tao; Xiao, Run-mei; Wang, Ming-feng; Wang, Jun-jun; Chen, Yong

    2016-01-01

    The regulation mechanism of arecoline on rat hepatic CYP2E1 was studied in vivo. After oral administration of arecoline hydrobromide (AH; 4, 20 and 100 mg x kg(-1) x d(-1)) to rats for one week, the hepatic CYP2E1 mRNA level remained unchanged, but the hepatic CYP2E1 protein content was dose-dependently increased. Additionally, although the hepatic CYP2E1 activity was induced by AH treatment, the induction was attenuated with the increase in dosage. The results indicate that the effect of arecoline on rat hepaticdoes not involve transcriptional activation of the gene, but largely involves the stabilization of CYP2E1 protein against degradation or increased efficiency of CYP2E1 mRNA translation, and additionally involve the post- ranslational modification of CYP2E1 protein. Furthermore, the CYP2E1 response is fairly equal among the different species, the induction of rat hepatic CYP2E1 by arecoline suggests that there is a risk of metabolic interaction among the substrate drugs of CYP2E1 in betel-quid use human. PMID:27405178

  2. 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.

  3. Crew Strength Training

    NASA Video Gallery

    Train to develop your upper and lower body strength in your muscles and bones by performing body-weight squats and push-ups.The Train Like an Astronaut project uses the excitement of exploration to...

  4. Developing Strengths in Families

    ERIC Educational Resources Information Center

    Bowman, Ted

    1976-01-01

    There are few descriptions of growth experiences for total families. This paper describes one such model. It expresses the conviction that families need opportunities to come together with other families to identify strengths, sharpen communication skills, and establish goals. (Author)

  5. 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

  6. The frequency of cytochrome P450 2E1 polymorphisms in Black South Africans.

    PubMed

    Chelule, Paul K; Pegoraro, Rosemary J; Gqaleni, Nceba; Dutton, Michael F

    2006-01-01

    Polymorphisms in the promoter region of the Cytochrome P4502E1 (CYP2E1) gene reportedly modify the metabolic activity of CYP2E1 enzyme, and have been associated with increased susceptibility to squamous cell carcinoma (SCC) of the oesophagus in high prevalence areas such as China. To assess the frequency of these polymorphisms in Black South Africans, a population with a high incidence of oesophageal SCC, this study examined genomic DNA from 331 subjects for restriction fragment length polymorphisms in the CYP2E1 (RsaI and PstI digestion). The frequency of the CYP2E1 c1/c1 and c1/c3 genotypes was 95% and 5% respectively. The frequency of the CYP2E1 allele distribution was found to be markedly different between Chinese and South African populations; hence it is important to place racial differences into consideration when proposing allelic variants as genetic markers for cancer. PMID:17264406

  7. Largazole and Its Derivatives Selectively Inhibit Ubiquitin Activating Enzyme (E1)

    PubMed Central

    Nasveschuk, Christopher G.; Wang, Wei; Quade, Bettina; Zhang, Gan; Kuchta, Robert D.; Phillips, Andrew J.; Liu, Xuedong

    2012-01-01

    Protein ubiquitination plays an important role in the regulation of almost every aspect of eukaryotic cellular function; therefore, its destabilization is often observed in most human diseases and cancers. Consequently, developing inhibitors of the ubiquitination system for the treatment of cancer has been a recent area of interest. Currently, only a few classes of compounds have been discovered to inhibit the ubiquitin-activating enzyme (E1) and only one class is relatively selective in E1 inhibition in cells. We now report that Largazole and its ester and ketone analogs selectively inhibit ubiquitin conjugation to p27Kip1 and TRF1 in vitro. The inhibitory activity of these small molecules on ubiquitin conjugation has been traced to their inhibition of the ubiquitin E1 enzyme. To further dissect the mechanism of E1 inhibition, we analyzed the effects of these inhibitors on each of the two steps of E1 activation. We show that Largazole and its derivatives specifically inhibit the adenylation step of the E1 reaction while having no effect on thioester bond formation between ubiquitin and E1. E1 inhibition appears to be specific to human E1 as Largazole ketone fails to inhibit the activation of Uba1p, a homolog of E1 in Schizosaccharomyces pombe. Moreover, Largazole analogs do not significantly inhibit SUMO E1. Thus, Largazole and select analogs are a novel class of ubiquitin E1 inhibitors and valuable tools for studying ubiquitination in vitro. This class of compounds could be further developed and potentially be a useful tool in cells. PMID:22279528

  8. 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...

  9. 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

  10. 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.

  11. Expression and characterization of a soluble rubella virus E1 envelope protein.

    PubMed

    Seto, N O; Gillam, S

    1994-10-01

    Individual specific antigenic rubella virus (RV) structural proteins are required for accurate serological diagnosis of acute and congenital rubella infections as well as rubella immune status. The RV envelope glycoprotein E1 is the major target antigen and plays an important role in viral-specific immune responses. The native virion is difficult to produce in large quantities and the protein subunits are also difficult to isolate without loss of antigenicity. The production of a soluble RV E1 (designated E1 delta Tm) using the baculovirus-insect cell expression system is described. In contrast to wild-type RV E1, the genetically engineered E1 delta Tm protein lacks a transmembrane anchor. It behaved as a secretory protein and was secreted abundantly from insect cells. Pulse-chase studies were used to examine the synthesis, glycosylation, and secretion of E1 delta Tm by the insect cells. The secreted E1 delta Tm protein was purified from serum-free medium by one-step immunochromatography. The purified E1 delta Tm protein retained full antigenicity and may be a convenient source of E1 protein for use in diagnostic assay and rubella vaccine development. PMID:7852960

  12. HPV E1 up-regulates replication-related biochemistries of AAV Rep78.

    PubMed

    Bandyopadhyay, Sarmistha; Cao, Maohua; Liu, Yong; Hermonat, Paul L

    2010-06-20

    Human papillomavirus type 16 (HPV) E1 protein provides helper function for the adeno-associated virus type 2 (AAV) life cycle. E1 is the replication protein of HPV, analogous to AAV Rep78, but without the endonuclease/covalent attachment activity of Rep78. Previously we have shown that E1 and Rep78 interact in vitro. Here we investigated E1's effects on Rep78 interaction with AAV's inverted terminal repeat (ITR) DNA in vitro, using purified Rep78 and E1 proteins from bacteria. E1 enhanced Rep78-ITR binding, ATPase activity, Rep78-ITR-covalent linkage and Rep78-ITR-endonuclease activity (central to AAV replication). These enhancements occurred in a dose-dependent manner whenever assayed. However, overall Rep78-plus-E1 helicase activity was lower than Rep78's helicase activity. These data suggest that E1's broad-based helper function for the AAV life cycle (AAV DNA, mRNA, and protein levels are up-regulated by E1) is likely through its ability to enhance Rep78's critical replication-required biochemistries on ITR DNA.

  13. Cell(MC3T3-E1)-printed poly(ϵ-caprolactone)/alginate hybrid scaffolds for tissue regeneration.

    PubMed

    Lee, Hyeongjin; Ahn, SeungHyun; Bonassar, Lawrence J; Kim, GeunHyung

    2013-01-25

    A new cell-printed scaffold consisting of poly(ϵ-caprolactone) (PCL) and cell-embedded alginate struts is designed. The PCL and alginate struts are stacked in an interdigitated pattern in successive layers to acquire a three-dimensional (3D) shape. The hybrid scaffold exhibits a two-phase structure consisting of cell (MC3T3-E1)-laden alginate struts able to support biological activity and PCL struts able to provide controllable mechanical support of the cell-laden alginate struts. The hybrid scaffolds exhibit an impressive increase in tensile modulus and maximum strength compared to pure alginate scaffolds. Laden cells are homogeneously distributed throughout the alginate struts and the entire scaffold, resulting in cell viability of approximately 84%.

  14. Reduction of benzene metabolism and toxicity in mice that lack CYP2E1 expression.

    PubMed

    Valentine, J L; Lee, S S; Seaton, M J; Asgharian, B; Farris, G; Corton, J C; Gonzalez, F J; Medinsky, M A

    1996-11-01

    Transgenic CYP2E1 knockout mice (cyp2e1-/-) were used to investigate the involvement of CYP2E1 in the in vivo metabolism of benzene and in the development of benzene-induced toxicity. After benzene exposure, absence of CYP2E1 protein was confirmed by Western blot analysis of mouse liver samples. For the metabolism studies, male cyp2e1-/- and wild-type control mice were exposed to 200 ppm benzene, along with a radiolabeled tracer dose of [14C]benzene (1.0 Ci/mol) by nose-only inhalation for 6 hr. Total urinary radioactivity and all radiolabeled individual metabolites were reduced in urine of cyp2e1-/- mice compared to wild-type controls during the 48-hr period after benzene exposure. In addition, a significantly greater percentage of total urinary radioactivity could be accounted for as phenylsulfate conjugates in cyp2e1-/- mice compared to wild-type mice, indicating the importance of CYP2E1 in oxidation of phenol following benzene exposure in normal mice. For the toxicity studies, male cyp2e1-/-, wild-type, and B6C3F1 mice were exposed by whole-body inhalation to 0 ppm (control) or 200 ppm benzene, 6 hr/day for 5 days. On Day 5, blood, bone marrow, thymus, and spleen were removed for evaluation of micronuclei frequencies and tissue cellularities. No benzene-induced cytotoxicity or genotoxicity was observed in cyp2e1-/- mice. In contrast, benzene exposure resulted in severe genotoxicity and cytotoxicity in both wild-type and B6C3F1 mice. These studies conclusively demonstrate that CYP2E1 is the major determinant of in vivo benzene metabolism and benzene-induced myelotoxicity in mice.

  15. Sex steroid hormones regulate constitutive expression of Cyp2e1 in female mouse liver

    PubMed Central

    Cheng, Jie; Gonzalez, Frank J.

    2013-01-01

    CYP2E1 is of paramount toxicological significance because it metabolically activates a large number of low-molecular-weight toxicants and carcinogens. In this context, factors that interfere with Cyp2e1 regulation may critically affect xenobiotic toxicity and carcinogenicity. The aim of this study was to investigate the role of female steroid hormones in the regulation of CYP2E1, as estrogens and progesterone are the bases of contraceptives and hormonal replacement therapy in menopausal women. Interestingly, a fluctuation in the hepatic expression pattern of Cyp2e1 was revealed in the different phases of the estrous cycle of female mice, with higher Cyp2e1 expression at estrus (E) and lower at methestrus (ME), highly correlated with that in plasma gonadal hormone levels. Depletion of sex steroids by ovariectomy repressed Cyp2e1 expression to levels similar to those detected in males and cyclic females at ME. Hormonal supplementation brought Cyp2e1 expression back to levels detected at E. The role of progesterone appeared to be more prominent than that of 17β-estradiol. Progesterone-induced Cyp2e1 upregulation could be attributed to inactivation of the insulin/PI3K/Akt/FOXO1 signaling pathway. Tamoxifen, an anti-estrogen, repressed Cyp2e1 expression potentially via activation of the PI3K/Akt/FOXO1 and GH/STAT5b-linked pathways. The sex steroid hormone-related changes in hepatic Cyp2e1 expression were highly correlated with those observed in Hnf-1α, β-catenin, and Srebp-1c. In conclusion, female steroid hormones are clearly involved in the regulation of CYP2E1, thus affecting the metabolism of a plethora of toxicants and carcinogenic agents, conditions that may trigger several pathologies or exacerbate the outcomes of various pathophysiological states. PMID:23548611

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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...

  1. 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...

  2. 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...

  3. 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...

  4. 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...

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

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Section 52e.1 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS NATIONAL 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...

  6. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Section 52e.1 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS NATIONAL 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...

  7. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Section 52e.1 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS NATIONAL 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...

  8. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Section 52e.1 Public Health PUBLIC HEALTH SERVICE, DEPARTMENT OF HEALTH AND HUMAN SERVICES GRANTS NATIONAL 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...

  9. 26 CFR 1.665(e)-1A - Preceding taxable year.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 8 2014-04-01 2014-04-01 false Preceding taxable year. 1.665(e)-1A Section 1... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning on Or After January 1, 1969 § 1.665(e)-1A Preceding taxable year. (a) Definition—(1)...

  10. 26 CFR 1.665(e)-1A - Preceding taxable year.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 8 2012-04-01 2012-04-01 false Preceding taxable year. 1.665(e)-1A Section 1... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning on Or After January 1, 1969 § 1.665(e)-1A Preceding taxable year. (a) Definition—(1)...

  11. 26 CFR 1.665(e)-1A - Preceding taxable year.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 8 2011-04-01 2011-04-01 false Preceding taxable year. 1.665(e)-1A Section 1... (CONTINUED) INCOME TAXES (CONTINUED) Treatment of Excess Distributions of Trusts Applicable to Taxable Years Beginning on Or After January 1, 1969 § 1.665(e)-1A Preceding taxable year. (a) Definition—(1)...

  12. Resolvin E1 and chemokine-like receptor 1 mediate bone preservation.

    PubMed

    Gao, Li; Faibish, Dan; Fredman, Gabrielle; Herrera, Bruno S; Chiang, Nan; Serhan, Charles N; Van Dyke, Thomas E; Gyurko, Robert

    2013-01-15

    The polyunsaturated ω-3 fatty acid eicosapentaenoic acid-derived resolvin E1 (RvE1) enhances resolution of inflammation, prevents bone loss, and induces bone regeneration. Although the inflammation-resolving actions of RvE1 are characterized, the molecular mechanism of its bone-protective actions are of interest. To test the hypothesis that receptor-mediated events impact bone changes, we prepared transgenic mice overexpressing the RvE1 receptor chemokine-like receptor 1 (chemR23) on leukocytes. In zymosan-initiated peritonitis, neutrophil polymorphonuclear leukocyte infiltration in response to RvE1 was limited requiring log order lower doses in chemR23tg mice. Ligature-induced alveolar bone loss was diminished in chemR23tg mice. Local RvE1 treatment of uniform craniotomy in the parietal bone significantly accelerated regeneration of the bone defect. In in vitro bone cultures, RvE1 significantly enhanced expression of osteoprotegerin (OPG) without inducing change in receptor activator of NF-κB ligand levels, whereas the osteogenic markers alkaline phosphatase, bone sialoprotein, and Runt-related transcription factor 2 remained unchanged. These results indicate that RvE1 modulates osteoclast differentiation and bone remodeling by direct actions on bone, rescuing OPG production and restoring a favorable receptor activator of NF-κB ligand/OPG ratio, in addition to known anti-inflammatory and proresolving actions. PMID:23241890

  13. 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 §...

  14. 26 CFR 31.3121(e)-1 - State, United States, and citizen.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 15 2013-04-01 2013-04-01 false State, United States, and citizen. 31.3121(e)-1... § 31.3121(e)-1 State, United States, and citizen. (a) When used in the regulations in this subpart, the..., the term “United States”, when used in a geographical sense, means the several states (including...

  15. 26 CFR 31.3121(e)-1 - State, United States, and citizen.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 15 2010-04-01 2010-04-01 false State, United States, and citizen. 31.3121(e)-1... § 31.3121(e)-1 State, United States, and citizen. (a) When used in the regulations in this subpart, the..., the term “United States”, when used in a geographical sense, means the several states (including...

  16. 26 CFR 1.509(e)-1 - Definition of gross investment income.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 7 2013-04-01 2013-04-01 false Definition of gross investment income. 1.509(e)-1 Section 1.509(e)-1 Internal Revenue INTERNAL REVENUE SERVICE, DEPARTMENT OF THE TREASURY... gross investment income. For the distinction between gross receipts and gross investment income, see §...

  17. 77 FR 42677 - Special Conditions: General Electric CT7-2E1 Turboshaft Engine

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-20

    ... Federal Aviation Administration 14 CFR Part 33 Special Conditions: General Electric CT7-2E1 Turboshaft.... SUMMARY: This action proposes special conditions for the General Electric CT7-2E1 engine model. This.... Background On September 10, 2009, General Electric applied for an amendment to type certificate E8NE to...

  18. 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

  19. 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.

  20. 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

  1. 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

  2. Beyond Oncolytics: E1B55K-Deleted Adenovirus as a Vaccine Delivery Vector

    PubMed Central

    Thomas, Michael A.; Nyanhete, Tinashe; Tuero, Iskra; Venzon, David; Robert-Guroff, Marjorie

    2016-01-01

    Type 5 human adenoviruses (Ad5) deleted of genes encoding the early region 1B 55-kDa (E1B55K) protein including Onyx-015 (dl1520) and H101 are best known for their oncolytic potential. As a vaccine vector the E1B55K deletion may allow for the insertion of a transgene nearly 1,000 base pairs larger than now possible. This has the potential of extending the application for which the vectors are clinically known. However, the immune priming ability of E1B55K-deleted vectors is unknown, undermining our ability to gauge their usefulness in vaccine applications. For this reason, we created an E1B55K-deleted Ad5 vector expressing full-length single chain HIVBaLgp120 attached to a flexible linker and the first two domains of rhesus CD4 (rhFLSC) in exchange for the E3 region. In cell-based experiments the E1B55K-deleted vector promoted higher levels of innate immune signals including chemokines, cytokines, and the NKG2D ligands MIC A/B compared to an E1B55K wild-type vector expressing the same immunogen. Based on these results we evaluated the immune priming ability of the E1B55K-deleted vector in mice. The E1B55K-deleted vector promoted similar levels of Ad5-, HIVgp120, and rhFLSC-specific cellular and humoral immune responses as the E1B55K wild-type vector. In pre-clinical HIV-vaccine studies the wild-type vector has been employed as part of a very effective prime-boost strategy. This study demonstrates that E1B55K-deleted adenoviruses may serve as effective vaccine delivery vectors. PMID:27391605

  3. Beyond Oncolytics: E1B55K-Deleted Adenovirus as a Vaccine Delivery Vector.

    PubMed

    Thomas, Michael A; Nyanhete, Tinashe; Tuero, Iskra; Venzon, David; Robert-Guroff, Marjorie

    2016-01-01

    Type 5 human adenoviruses (Ad5) deleted of genes encoding the early region 1B 55-kDa (E1B55K) protein including Onyx-015 (dl1520) and H101 are best known for their oncolytic potential. As a vaccine vector the E1B55K deletion may allow for the insertion of a transgene nearly 1,000 base pairs larger than now possible. This has the potential of extending the application for which the vectors are clinically known. However, the immune priming ability of E1B55K-deleted vectors is unknown, undermining our ability to gauge their usefulness in vaccine applications. For this reason, we created an E1B55K-deleted Ad5 vector expressing full-length single chain HIVBaLgp120 attached to a flexible linker and the first two domains of rhesus CD4 (rhFLSC) in exchange for the E3 region. In cell-based experiments the E1B55K-deleted vector promoted higher levels of innate immune signals including chemokines, cytokines, and the NKG2D ligands MIC A/B compared to an E1B55K wild-type vector expressing the same immunogen. Based on these results we evaluated the immune priming ability of the E1B55K-deleted vector in mice. The E1B55K-deleted vector promoted similar levels of Ad5-, HIVgp120, and rhFLSC-specific cellular and humoral immune responses as the E1B55K wild-type vector. In pre-clinical HIV-vaccine studies the wild-type vector has been employed as part of a very effective prime-boost strategy. This study demonstrates that E1B55K-deleted adenoviruses may serve as effective vaccine delivery vectors. PMID:27391605

  4. 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.

  5. Notch strength of composites

    NASA Technical Reports Server (NTRS)

    Whitney, J. M.

    1983-01-01

    The notch strength of composites is discussed. The point stress and average stress criteria relate the notched strength of a laminate to the average strength of a relatively long tensile coupon. Tests of notched specimens in which microstrain gages have been placed at or near the edges of the holes have measured strains much larger that those measured in an unnotched tensile coupon. Orthotropic stress concentration analyses of failed notched laminates have also indicated that failure occurred at strains much larger than those experienced on tensile coupons with normal gage lengths. This suggests that the high strains at the edge of a hole can be related to the very short length of fiber subjected to these strains. Lockheed has attempted to correlate a series of tests of several laminates with holes ranging from 0.19 to 0.50 in. Although the average stress criterion correlated well with test results for hole sizes equal to or greater than 0.50 in., it over-estimated the laminate strength in the range of hole sizes from 0.19 to 0.38 in. It thus appears that a theory is needed that is based on the mechanics of failure and is more generally applicable to the range of hole sizes and the varieties of laminates found in aircraft construction.

  6. 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.

  7. Gender Differences in Strength.

    ERIC Educational Resources Information Center

    Heyward, Vivian H.; And Others

    1986-01-01

    This investigation examined gender differences of 103 physically active men and women in upper and lower body strength as a function of lean body weight and the distribution of muscle and subcutaneous fat in the upper and lower limbs. Results are discussed. (Author/MT)

  8. 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

  9. Systematics of low-lying E1 and M1 excitations in heavy nuclei from photon scattering experiments

    NASA Astrophysics Data System (ADS)

    Pitz, H. H.; Bauwens, F.; Belic, D.; Bryssinck, J.; von Brentano, P.; Fransen, C.; De Frenne, D.; Govor, L.; Herzberg, R.-D.; Jacobs, E.; Kneissl, U.; Kohstall, C.; Linnemann, A.; Maser, H.; Matschinsky, P.; Nord, A.; Pietralla, N.; Ponomarev, V. Yu.; Scheck, M.; Stedile, F.; Werner, V.

    2000-07-01

    The results of systematic nuclear resonance fluorescence experiments on heavy deformed and spherical nuclei at the bremsstrahlung facility of the Stuttgart Dynamitron accelerator are reported. The investigation of the fragmentation of the orbital M1 scissors mode in deformed odd-mass nuclei has been continued with high-precision measurements on 166Ho and 163Dy, revealing a lot of weak excitations distributed over the energy range 1.5-4 MeV. The investigation of the (2+⊗3-) two-phonon excitations in the Sn isotopic chain shows a very harmonic coupling of the phonons. The results for both, the even-even and the odd-mass Sn isotopes are in good agreement with QPM calculations.

  10. Resolvin E1 Inhibits Substance P-Induced Potentiation of TRPV1 in Primary Sensory Neurons

    PubMed Central

    Jo, Youn Yi; Lee, Ji Yeon

    2016-01-01

    The neuropeptide substance P (SP) is expressed in primary sensory neurons and is commonly regarded as a “pain” neurotransmitter. Upon peripheral inflammation, SP activates the neurokinin-1 (NK-1) receptor and potentiates activity of transient receptor potential vanilloid subtype 1 (TRPV1), which is coexpressed by nociceptive neurons. Therefore, SP functions as an important neurotransmitter involved in the hypersensitization of inflammatory pain. Resolvin E1 (RvE1), derived from omega-3 polyunsaturated fatty acids, inhibits TRPV1 activity via activation of the chemerin 23 receptor (ChemR23)—an RvE1 receptor located in dorsal root ganglion neurons—and therefore exerts an inhibitory effect on inflammatory pain. We demonstrate here that RvE1 regulates the SP-induced potentiation of TRPV1 via G-protein coupled receptor (GPCR) on peripheral nociceptive neurons. SP-induced potentiation of TRPV1 inhibited by RvE1 was blocked by the Gαi-coupled GPCR inhibitor pertussis toxin and the G-protein inhibitor GDPβ-S. These results indicate that a low concentration of RvE1 strongly inhibits the potentiation of TRPV1, induced by the SP-mediated activation of NK-1, via a GPCR signaling pathway activated by ChemR23 in nociceptive neurons. RvE1 might represent a new therapeutic target for the treatment of inflammatory pain as a prospective endogenous inhibitor that strongly inhibits TRPV1 activity associated with peripheral inflammation. PMID:27738388

  11. Characterization of in vivo functions of Nicotiana benthamiana RabE1.

    PubMed

    Ahn, Chang Sook; Han, Jeong-A; Pai, Hyun-Sook

    2013-01-01

    We characterized the gene expression, subcellular localization, and in vivo functions of a Nicotiana benthamiana small GTPase belonging to the RabE family, designated NbRabE1. The NbRabE1 promoter drove strong β-glucuronidase reporter expression in young tissues containing actively dividing cells and in stomata guard cells. GFP fusion proteins of NbRabE1 and its dominant-negative and constitutively active mutants were all localized to the Golgi apparatus and the plasma membrane but showed different affinities for membrane attachment. Virus-induced gene silencing of NbRabE1 resulted in pleiotropic phenotypes, including growth arrest, premature senescence, and abnormal leaf development. At the cellular level, the leaves in which NbRabE1 was silenced contained abnormal stomata that lacked pores or contained incomplete ventral walls, suggesting that NbRabE1 deficiency leads to defective guard cell cytokinesis. Ectopic expression of the dominant-negative mutant of NbRabE1 in Arabidopsis thaliana resulted in retardation of shoot and root growth accompanied by defective root hair formation. These developmental defects are discussed in conjunction with proposed functions of RabE GTPases in polarized secretory vesicle trafficking.

  12. 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

  13. 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.

  14. The pyruvate dehydrogenase E1 alpha gene is testosterone and prolactin regulated in prostate epithelial cells.

    PubMed

    Costello, L C; Liu, Y; Zou, J; Franklin, R B

    2000-02-01

    The prostate gland of humans and other animals has the unique function of accumulating and secreting extraordinarily high levels of citrate. The prostate secretory epithelial cells synthesize citrate which, due to a limiting mitochondrial (m-) aconitase, accumulates rather than being oxidized. Thus citrate is essentially an end product of metabolism in prostate. For continued net citrate production, a continual source of oxaloacetate (OAA) and acetyl CoA is required. Glucose via pyruvate oxidation provides the source of Acetyl CoA. In prostate cells, citrate production is regulated by testosterone and/or by prolactin. Both hormones selectively regulate the level and activity of pyruvate dehydrogenase E1 alpha (E1a) in animal prostate cells; thereby regulating the availability of acetyl CoA for citrate synthesis. Studies were conducted to determine if testosterone and prolactin might regulate the expression of the E1a gene in prostate epithelial cells. Prolactin treatment of rat ventral and lateral prostate cells and human PC3 cells increased the levels of E1a mRNA and the rates of transcription of the E1a gene. Testosterone also increased the mRNA level and transcription of E1a in rat ventral prostate cells, and in PC3 cells transfected with androgen receptor. However, testosterone treatment resulted in a repression of E1a gene expression in lateral prostate cells. Evidence is presented which supports the view that prolactin regulation of E1a is mediated via PKC. The rapidity of the effects of both hormones is representative of an immediate-early gene response. To our knowledge this represents the first report in any mammalian cells that, in addition to its constitutive expression in all mammalian cells, the E1a gene is a hormonally-regulated gene in specifically targeted prostate epithelial cells. PMID:10711720

  15. 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.

  16. Corium crust strength measurements.

    SciTech Connect

    Lomperski, S.; Nuclear Engineering Division

    2009-11-01

    Corium strength is of interest in the context of a severe reactor accident in which molten core material melts through the reactor vessel and collects on the containment basemat. Some accident management strategies involve pouring water over the melt to solidify it and halt corium/concrete interactions. The effectiveness of this method could be influenced by the strength of the corium crust at the interface between the melt and coolant. A strong, coherent crust anchored to the containment walls could allow the yet-molten corium to fall away from the crust as it erodes the basemat, thereby thermally decoupling the melt from the coolant and sharply reducing the cooling rate. This paper presents a diverse collection of measurements of the mechanical strength of corium. The data is based on load tests of corium samples in three different contexts: (1) small blocks cut from the debris of the large-scale MACE experiments, (2) 30 cm-diameter, 75 kg ingots produced by SSWICS quench tests, and (3) high temperature crusts loaded during large-scale corium/concrete interaction (CCI) tests. In every case the corium consisted of varying proportions of UO{sub 2}, ZrO{sub 2}, and the constituents of concrete to represent a LWR melt at different stages of a molten core/concrete interaction. The collection of data was used to assess the strength and stability of an anchored, plant-scale crust. The results indicate that such a crust is likely to be too weak to support itself above the melt. It is therefore improbable that an anchored crust configuration could persist and the melt become thermally decoupled from the water layer to restrict cooling and prolong an attack of the reactor cavity concrete.

  17. Strength calculations on airplanes

    NASA Technical Reports Server (NTRS)

    Baumann, A

    1925-01-01

    Every strength calculation, including those on airplanes, must be preceded by a determination of the forces to be taken into account. In the following discussion, it will be assumed that the magnitudes of these forces are known and that it is only a question of how, on the basis of these known forces, to meet the prescribed conditions on the one hand and the practical requirements on the other.

  18. The C-terminal region of E1A: a molecular tool for cellular cartography.

    PubMed

    Yousef, Ahmed F; Fonseca, Gregory J; Cohen, Michael J; Mymryk, Joe S

    2012-04-01

    The adenovirus E1A proteins function via protein-protein interactions. By making many connections with the cellular protein network, individual modules of this virally encoded hub reprogram numerous aspects of cell function and behavior. Although many of these interactions have been thoroughly studied, those mediated by the C-terminal region of E1A are less well understood. This review focuses on how this region of E1A affects cell cycle progression, apoptosis, senescence, transformation, and conversion of cells to an epithelial state through interactions with CTBP1/2, DYRK1A/B, FOXK1/2, and importin-α. Furthermore, novel potential pathways that the C-terminus of E1A influences through these connections with the cellular interaction network are discussed.

  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. 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

  1. DprE1--from the discovery to the promising tuberculosis drug target.

    PubMed

    Mikusová, Katarína; Makarov, Vadim; Neres, João

    2014-01-01

    Several groups working in the field of the development of new antituberculosis drugs have recently reported active compounds targeting mycobacterial enzyme DprE1. Along with its counterpart, DprE2, it catalyses a unique epimerization reaction resulting in the synthesis of decaprenylphosphoryl arabinose, the single donor of arabinosyl residues for the build-up of arabinans, fundamental components of the mycobacterial cell wall. This review presents the historical background leading to the discovery of DprE1, focusing on the biochemical and structural characterization of this important emerging target and introducing the molecules acting on DprE1 including the development of the most successful series--the benzothiazinones, currently in late pre-clinical development, which turned to be suicide inhibitors of DprE1.

  2. The E1-E2 center in gallium arsenide is the divacancy.

    PubMed

    Schultz, Peter A

    2015-02-25

    Based on defect energy levels computed from first-principles calculations, it is shown the E1-E2 center in irradiated GaAs cannot be due to an isolated arsenic vacancy. The only simple intrinsic defect with levels compatible with E1 and E2 is the divacancy. The arsenic monovacancy is reassigned to the E3 center in irradiated GaAs. These new assignments are shown to reconcile a number of seemingly contradictory experimental observations.

  3. Impact of resolvin E1 on murine neutrophil phagocytosis in type 2 diabetes.

    PubMed

    Herrera, Bruno S; Hasturk, Hatice; Kantarci, Alpdogan; Freire, Marcelo O; Nguyen, Olivia; Kansal, Shevali; Van Dyke, Thomas E

    2015-02-01

    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.

  4. Cisplatin-induced hepatotoxicity is enhanced by elevated expression of cytochrome P450 2E1.

    PubMed

    Lu, Yongke; Cederbaum, Arthur I

    2006-02-01

    In this study, the possible potentiation of cisplatin-induced hepatotoxicity by cytochrome P450 2E1 (CYP2E1) was examined both in vitro and in vivo. Transfected HepG2 cells expressing CYP2E1 (E47 cells) and not expressing CYP2E1 (C34 cells) were used as an in vitro model, and mice drinking 2% acetone for 7 days to induce CYP2E1 were used as an in vivo model. Exposure of E47 cells to cisplatin caused a much greater loss of cell viability, more striking depletion of reduced glutathione (GSH), and higher reactive oxygen species (ROS) production as compared with C34 cells. The prooxidant L-buthionine-[R,S]-sulfoximine (BSO), which depletes GSH, enhanced cisplatin-induced loss of cell viability, whereas the antioxidant glutathione ethyl ester, or the iron chelator deferoxamine mesylate (DFO) protected against the cisplatin-induced loss of E47 cell viability. Diallyl sulfide (DAS), an inhibitor of CYP2E1, also protected against the cisplatin toxicity in the E47 cells. After being injected with cisplatin (ip, 45 mg/kg), mice drinking 2% acetone with increased CYP2E1 levels exhibited elevated levels of serum ALT and AST, liver caspase-3 activity and positive staining of TUNEL increased, and histopathology indicated the presence of necrotic foci in livers of acetone plus cisplatin-treated mice. Lipid peroxidation and protein oxidation as indicated by carbonyl formation, staining of 3-nitrotyrosine (3-NT) and iron were higher in the cisplatin plus acetone group, compared with cisplatin alone group. Both in vitro and in vivo results indicate that elevated CYP2E1 enhances cisplatin-induced hepatotoxicity, and the mechanism may involve increased production of ROS and oxidative stress.

  5. 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.

  6. 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

  7. Resolvin D1 and E1 promote resolution of inflammation in microglial cells in vitro.

    PubMed

    Rey, C; Nadjar, A; Buaud, B; Vaysse, C; Aubert, A; Pallet, V; Layé, S; Joffre, C

    2016-07-01

    Sustained inflammation in the brain together with microglia activation can lead to neuronal damage. Hence limiting brain inflammation and activation of microglia is a real therapeutic strategy for inflammatory disease. Resolvin D1 (RvD1) and resolvin E1 (RvE1) derived from n-3 long chain polyunsaturated fatty acids are promising therapeutic compounds since they actively turn off the systemic inflammatory response. We thus evaluated the anti-inflammatory activities of RvD1 and RvE1 in microglia cells in vitro. BV2 cells were pre-incubated with RvD1 or RvE1 before lipopolysaccharide (LPS) treatment. RvD1 and RvE1 both decreased LPS-induced proinflammatory cytokines (TNF-α, IL-6 and IL-1β) gene expression, suggesting their proresolutive activity in microglia. However, the mechanisms involved are distinct as RvE1 regulates NFκB signaling pathway and RvD1 regulates miRNAs expression. Overall, our findings support that pro-resolving lipids are involved in the resolution of brain inflammation and can be considered as promising therapeutic agents for brain inflammation. PMID:26718448

  8. RNA1 is sufficient to mediate plasmid ColE1 incompatibility in vivo.

    PubMed

    Fitzwater, T; Tamm, J; Polisky, B

    1984-05-25

    The multicopy plasmid ColE1 specifies a small RNA designated RNA1 that has been implicated in copy number control and incompatibility. We have inserted a 148 base-pair ColE1 DNA fragment containing a promoter-less RNA1 gene into a plasmid vector downstream from the tryptophan promoter of Serratia marcesens . The ColE1 RNA1 produced by this plasmid is not functional in vivo due to the presence of 49 nucleotides appended to the 5'-terminus of the wild-type RNA1 sequence. Deletions of these sequences by Bal3l nuclease in vitro and genetic selection for ColE1 incompatibility function in vivo permitted isolation of a plasmid expressing wild-type ColE1 RNA1 initiated properly from the S. marcesens trp promoter. These experiments demonstrate that RNA1 is sufficient to mediate ColE1 incompatibility in vivo. In addition, several plasmids were isolated that contain altered RNA1 genes. These alterations consist of additions or deletions of sequences at the 5'-terminus of RNA1. Analysis of the ability of these altered RNA1 molecules to express incompatibility in vivo suggests that the 5'-terminal region of RNA1 is crucial for its function.

  9. Response of an Escherichia coli-Bound Fluorescent Probe to Colicin E1

    PubMed Central

    Cramer, W. A.; Phillips, S. K.

    1970-01-01

    The fluorescent probe, 8-anilino-1-napthalenesulfonate (ANS) binds to Escherichia coli, showing an enhanced fluorescence. The interaction of colicin E1 with sensitive cells causes an increase of about 100% in the fluorescence of the bound ANS, and this change at equilibrium has an apparent “all-or-none” nature as a function of E1 multiplicity. Approximately 6 to 8% of the ANS is bound to the cells at equilibrium. The colicin E1-induced fluorescence increase can be attributed partly to an increase in ANS binding and partly to an increase in the fluorescence yield of the bound ANS. The kinetics of the E1-induced fluorescence increase in sensitive cells are very similar to those of the adenosine triphosphate decrease. The phosphorylation uncoupler p-trifluoromethoxy-carbonylcyanidephenylhydrazone also causes a large change in the fluorescence of bound ANS. Colicin E2 or E3 does not cause any fluorescence change, nor does colicin E1 cause fluorescence change with a colicinogenic strain. ANS appears to be a probe of structural or conformational change in the cell envelope that is closely associated with the colicin E1-induced adenosine triphosphate decrease. PMID:4923074

  10. Fragment E1 labeled with I-123 in the detection of venous thrombosis

    SciTech Connect

    Knight, L.C.; Maurer, A.H.; Robbins, P.S.; Malmud, L.S.; Budzynski, A.Z.

    1985-08-01

    Fragment E1, which has been shown to have specific binding affinity for thrombi in an animal model, was investigated in humans for its safety and ability to bind to venous thrombi. Human Fragment E1 was labeled with I-123 and administered intravenously to patients with proved or suspected deep vein thrombosis. The vascular distribution of radioactivity was documented by obtaining gamma camera images of the patients' legs for 30 minutes following administration of I-123-Fragment E1. All patients (n = 5) with documented venous thrombi had rapid localization of labeled Fragment E1 in the area of thrombus. Patients without evidence of thrombi (n = 5) showed no focal localization, although two of these patients showed diffuse uptake along the length of the veins, due to superficial phlebitis. Analysis of blood samples in four patients indicated that disappearance of Fragment E1 from the circulation was more rapid in individuals with thrombosis (t 1/2 = 20 min) than in individuals without thrombosis (t 1/2 = 90 min), and a radiolabeled species of high molecular weight was found in patients with thrombosis but was absent from patients without thrombosis. These early results suggest that radiolabeled Fragment E1 is a safe and potentially valuable agent for the rapid detection of venous thrombosis.

  11. Two domains within the adenovirus type 12 E1A unique spacer have disparate effects on the interaction of E1A with P105-Rb and the transformation of primary mouse cells.

    PubMed

    Rumpf, H; Esche, H; Kirch, H C

    1999-04-25

    Transformation of primary rodent cells by functions of the adenovirus type 12 (Ad12) early region 1 (E1) is reduced severalfold compared with transformation by E1 of Ad2. We analyzed whether the unique spacer region of Ad12 E1A that borders the conserved region (CR) 2 and represents an oncogenic determinant of Ad12 E1A is involved in this impaired transformation property, putatively by modulating transformation-relevant biological E1A functions. We show that a mutant (E1ASpm1) that lacks 12 amino-terminal residues of the spacer binds p105-Rb and p130 as Ad12 E1A wild type (E1Awt), whereas a second spacer mutant (E1ASpm2) that lacks an adjacent stretch of six alanines exhibits highly reduced binding to p105-Rb. The binding of this mutant to the p130 pocket protein is, however, little impaired. E1ASpm1 diminishes the formation of the p105-Rb-E2F complex more efficiently than E1Awt or, least efficient, E1ASpm2. These properties of the spacer mutants to target and to disintegrate the p105-Rb-E2F complex correspond with their ability to transform primary mouse cells in combination with E1B: E1ASpm1 (plus Ad12 E1B)-transfected cells could be easily established as cell lines, comparable to Ad12 E1Awt- or Ad2 E1Awt-transfected cells. In contrast, cells transfected with E1ASpm2 or Ad12 E1AdelCR2 (lacking the entire CR2) died within 6-10 weeks after replating, although foci were formed in all cases. Of note, the E1ASpm1-transformed cells grow as fast as the Ad2 E1Awt-transformed cells, with a doubling rate of 15 h, whereas the doubling of the Ad12 E1Awt-transformed cells takes approximately 120 h. Moreover, in the established cell lines, the affinity of E1ASpm1 to p105-Rb was higher than with that of E1Awt. Our data suggest the presence of a transformation-suppressing domain within the carboxyl-terminal 12 residues of the Ad12 E1A-unique spacer, whereas the hydrophobic stretch of six alanines in the spacer is required for stable transformation. PMID:10208919

  12. 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

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

    PubMed Central

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

    2013-01-01

    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 to investigate 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) (6 g/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 while liver histology obtained at 6 h 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-acetyl-cysteine, both of which suppressed the oxidative markers including intestinal CYP2E1. WT-EtOH also exhibited elevated amounts of serum TNF-α, 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 pro-apoptotic proteins and decreased levels of active (phosphorylated) p-AKT, p-AMPK and peroxisome proliferator-activated receptor-alpha (PPAR-α), 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 seem critical in the binge alcohol-mediated increased nitroxidative stress, gut leakage, endotoxemia, and

  14. 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-α, 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-α, 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

  15. Cytochrome p450 2E1 polymorphisms and the risk of gastric cardia cancer

    PubMed Central

    Cai, Lin; Zheng, Zong-Li; Zhang, Zuo-Feng

    2005-01-01

    AIM: Genetic polymorphisms of drug-metabolizing enzymes have recently been shown to affect susceptibility to chemical carcinogenesis. Cytochrome P450 2E1 (CYP2E1) enzyme catalyzes the metabolism of many procarcinogens, such as N-nitrosamines and related compounds. The gene coding for this enzyme is polymorphic and thus may play a role in gastric cardia cancer (GCC) etiology. In this hospital-based case-control study, we evaluate the relationship between genetic polymorphisms of CYP2E1 and the risk of GCC. METHODS: The study subjects comprised 159 histologically confirmed GCC cases identified via hospital cancer registry and surgical records at five hospitals in Fuzhou, Fujian Province, China, between April and November 2001. Controls were 192 patients admitted to the same hospitals for nonmalignant conditions. The genotypes of CYP2E1 were detected by a PCR-based RFLP assay. The odds ratios were estimated by logistic regression analyses and were adjusted for potential confounding factors. RESULTS: The distribution of three genotypes of CYP2E1 in GCC cases and controls was significantly different (χ2 = 16.04, P<0.01). The frequency of the CYP2E1 (c1/c1) genotype in GCC cases and controls was 60.4% and 40.1%, respectively. The CYP2E1 (c1/c1) genotype was associated with an increased risk for GCC (the adjusted (OR) was 2.37, 95% confidence interval (CI): 1.52-3.70). Subjects who carried the CYP2E1 (c1/c1) genotype and were habitual smokers were at a significantly higher risk of developing GCC (OR = 4.68, 95%CI: 2.19-10.04) compared with those who had the CYP2E1 (c1/c2 or c2/c2) genotype and did not smoke. CONCLUSION: These results suggest that the CYP2E1 genotype may influence individual susceptibility to development of GCC, and that the risk increases significantly in smokers. PMID:15793883

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

    PubMed

    Lu, Yongke; Cederbaum, Arthur I

    2015-10-16

    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

  17. Redox Regulation of Human Estrogen Sulfotransferase (hSULT1E1)†

    PubMed Central

    Maiti, Smarajit; Zhang, Jimei; Chen, Guangping

    2007-01-01

    Sulfotransferases (SULTs) are enzymes that catalyze the sulfation of hydroxyl-containing compounds. Sulfation regulates hormone activities and detoxifies xenobiotics. Human estrogen sulfotransferase (hSULT1E1) catalyzes the sulfation of estrogens and regulates estrogen bioactivities. Oxidative regulation provides a biological mechanism for regulating enzyme activities in vivo. The oxidative regulation of human SULTs has not been reported. In this study, we used amino acid modification, manipulation of intracellular redox state, and site-directed mutagenesis to study the redox regulation of human SULTs and specifically the mechanism of hSULT1E1 inhibitory regulation by oxidized glutathione (GSSG). Of the four major human SULTs, hSULT1A1, hSULT1A3, and hSULT2A1 do not undergo redox regulation; hSULT1E1, on the other hand, can be redox regulated. GSSG inactivated hSULT1E1 activity in an efficient, time- and concentration-dependant manner. The co-enzyme adenosine 3′-phosphate 5′-phosphosulfate protected hSULT1E1 from GSSG-associated inactivation. A reduced glutathione (GSH) inducer (N-acetyl cysteine) significantly increased while a GSH depletor (buthionine sulfoxamine) significantly decreased hSULT1E1 activity, but both failed to affect the amount of hSULT1E1 protein in human hepatocyte carcinoma Hep G2 cells. Crystal structure suggested that no Cys residues exist near the active sites of hSULT1A1, hSULT1A3, and hSULT2A1, but Cys residues do exist within the active site of hSULT1E1. Site-directed mutagenesis demonstrated that Cys83 is critical for the redox regulation of hSULT1E1. This first report on the redox regulation of human SULTs suggests that the redox regulation of hSULT1E1 may interrupt the regulation and function of estrogens under various physiological and pathological conditions. PMID:17266938

  18. Requirement for the E1 Helicase C-Terminal Domain in Papillomavirus DNA Replication In Vivo

    PubMed Central

    Bergvall, Monika; Gagnon, David; Titolo, Steve; Lehoux, Michaël; D'Abramo, Claudia M.

    2016-01-01

    ABSTRACT The papillomavirus (PV) E1 helicase contains a conserved C-terminal domain (CTD), located next to its ATP-binding site, whose function in vivo is still poorly understood. The CTD is comprised of an alpha helix followed by an acidic region (AR) and a C-terminal extension termed the C-tail. Recent biochemical studies on bovine papillomavirus 1 (BPV1) E1 showed that the AR and C-tail regulate the oligomerization of the protein into a double hexamer at the origin. In this study, we assessed the importance of the CTD of human papillomavirus 11 (HPV11) E1 in vivo, using a cell-based DNA replication assay. Our results indicate that combined deletion of the AR and C-tail drastically reduces DNA replication, by 85%, and that further truncation into the alpha-helical region compromises the structural integrity of the E1 helicase domain and its interaction with E2. Surprisingly, removal of the C-tail alone or mutation of highly conserved residues within the domain still allows significant levels of DNA replication (55%). This is in contrast to the absolute requirement for the C-tail reported for BPV1 E1 in vitro and confirmed here in vivo. Characterization of chimeric proteins in which the AR and C-tail from HPV11 E1 were replaced by those of BPV1 indicated that while the function of the AR is transferable, that of the C-tail is not. Collectively, these findings define the contribution of the three CTD subdomains to the DNA replication activity of E1 in vivo and suggest that the function of the C-tail has evolved in a PV type-specific manner. IMPORTANCE While much is known about hexameric DNA helicases from superfamily 3, the papillomavirus E1 helicase contains a unique C-terminal domain (CTD) adjacent to its ATP-binding site. We show here that this CTD is important for the DNA replication activity of HPV11 E1 in vivo and that it can be divided into three functional subdomains that roughly correspond to the three conserved regions of the CTD: an alpha helix, needed

  19. The E1A transcriptional control region is efficiently activated in proliferating tissues of transgenic mice.

    PubMed

    Dieckmann, A; Krippl, B

    1994-08-01

    To study the in vivo regulation of the adenovirus E1A transcriptional regulatory region in transgenic mice, we have constructed two hybrid genes in which the viral control element regulates the expression of the CAT and the lacZ reporter gene. The fusion constructs were introduced into the mouse germline. The expression of the transgenes were monitored during embryogenesis and during postnatal development as well as in adult organs. We show that the E1A regulatory region is recognized and activated in undifferentiated cells during early embryonic cleavage, in the morula, in the inner cell mass and in the trophectoderm of the blastocyst. Transcription initiation at the E1A promoter leads to higher marker gene expression in proliferative centers in postimplantation embryos at the beginning of the neural tube closure. Analysing marker gene expression during postnatal development, a correlation of transcriptional activity of the E1A regulatory region and cell proliferation could be demonstrated. The expression profile of the transgene in different adult organs parallels with DNA synthesis. Marker gene expression was high in cells of organs known to have a high mitotic rate, such as the intestine, the stomach, the skin and the bone marrow, whereas little activity of the E1A control region was observed in the post-proliferative brain. These results are consistent with the finding that activation of the viral cis-regulating elements dramatically increased in the kidney after mitotic stimulation by folic acid. These observations strongly suggests a cell cycle regulated expression from the E1A enhancer/promoter in the absence of the E1A autoregulatory proteins in the living animal. PMID:8036008

  20. Multifactorial comparative proteomic study of cytochrome P450 2E1 function in chronic alcohol administration.

    PubMed

    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.

  1. Mutation in E1, the ubiquitin activating enzyme, reduces Drosophila lifespan and results in motor impairment.

    PubMed

    Liu, Hsiu-Yu; Pfleger, Cathie M

    2013-01-01

    Neurodegenerative diseases cause tremendous suffering for those afflicted and their families. Many of these diseases involve accumulation of mis-folded or aggregated proteins thought to play a causal role in disease pathology. Ubiquitinated proteins are often found in these protein aggregates, and the aggregates themselves have been shown to inhibit the activity of the proteasome. These and other alterations in the Ubiquitin Pathway observed in neurodegenerative diseases have led to the question of whether impairment of the Ubiquitin Pathway on its own can increase mortality or if ongoing neurodegeneration alters Ubiquitin Pathway function as a side-effect. To address the role of the Ubiquitin Pathway in vivo, we studied loss-of-function mutations in the Drosophila Ubiquitin Activating Enzyme, Uba1 or E1, the most upstream enzyme in the Ubiquitin Pathway. Loss of only one functional copy of E1 caused a significant reduction in adult lifespan. Rare homozygous hypomorphic E1 mutants reached adulthood. These mutants exhibited further reduced lifespan and showed inappropriate Ras activation in the brain. Removing just one functional copy of Ras restored the lifespan of heterozygous E1 mutants to that of wild-type flies and increased the survival of homozygous E1 mutants. E1 homozygous mutants also showed severe motor impairment. Our findings suggest that processes that impair the Ubiquitin Pathway are sufficient to cause early mortality. Reduced lifespan and motor impairment are seen in the human disease X-linked Infantile Spinal Muscular Atrophy, which is associated with mutation in human E1 warranting further analysis of these mutants as a potential animal model for study of this disease.

  2. Increased Cytochrome P4502E1 Expression and Altered Hydroxyeicosatetraenoic Acid Formation Mediate Diabetic Vascular Dysfunction

    PubMed Central

    Schäfer, Andreas; Galuppo, Paolo; Fraccarollo, Daniela; Vogt, Christian; Widder, Julian D.; Pfrang, Julia; Tas, Piet; Barbosa-Sicard, Eduardo; Ruetten, Hartmut; Ertl, Georg; Fleming, Ingrid; Bauersachs, Johann

    2010-01-01

    OBJECTIVE We investigated the mechanisms underlying vascular endothelial and contractile dysfunction in diabetes as well as the effect of HMR1766, a novel nitric oxide (NO)-independent activator of soluble guanylyl cyclase (sGC). RESEARCH DESIGN AND METHODS Two weeks after induction of diabetes by streptozotocin, Wistar rats received either placebo or HMR1766 (10 mg/kg twice daily) for another 2 weeks; thereafter, vascular function was assessed. RESULTS Endothelial function and contractile responses were significantly impaired, while vascular superoxide formation was increased in the aortae from diabetic versus healthy control rats. Using RNA microarrays, cytochrome P4502E1 (CYP2E1) was identified as the highest upregulated gene in diabetic aorta. CYP2E1 protein was significantly increased (16-fold) by diabetes, leading to a reduction in levels of the potent vasoconstrictor 20-hydroxy-eicosatetraenoic acid (20-HETE). Induction of CYP2E1 expression in healthy rats using isoniazide mimicked the diabetic noncontractile vascular response while preincubation of aortae from STZ-diabetic rats in vitro with 20-HETE rescued contractile function. Chronic treatment with the sGC activator HMR1766 improved NO sensitivity and endothelial function, reduced CYP2E1 expression and superoxide formation, enhanced 20-HETE levels, and reversed the contractile deficit observed in the diabetic rats that received placebo. CONCLUSIONS Upregulation of CYP2E1 is essentially involved in diabetic vascular dysfunction. Chronic treatment with the sGC activator HMR1766 reduced oxidative stress, decreased CYP2E1 levels, and normalized vasomotor function in diabetic rats. PMID:20522591

  3. Dynamic Strength of Materials

    NASA Astrophysics Data System (ADS)

    Chhabildas, Lalit

    2011-06-01

    Historically when shock loading techniques became accessible in the early fifties it was assumed that materials behave like fluids implying that materials cannot support any shear stresses. Early and careful investigation in the sixties by G. R. Fowles in aluminum indicated otherwise. When he compared his Hugoniot compression measurements to hydrostatic pressure compression measurements in the pressure volume plane he noticed that the shock data lay above the hydrostatic compression curve - which laid the ground work for what is the basis for elastic-plastic theories that exist today. In this talk, a brief historical perspective on strength measurements in materials will be discussed including how time-resolved techniques have played a role in allowing estimates of the strength of materials at over Mbar stress. This is crucial especially at high stresses since we are determining values that are small compared to the loading stress. Even though we have made considerable progress in our understanding of materials, there are still many anomalies and unanswered questions. Some of these anomalies are fertile grounds for further and future research and will be mentioned.

  4. Oscillator strengths and collision strengths for S III

    NASA Technical Reports Server (NTRS)

    Ho, Y. K.; Henry, R. J. W.

    1984-01-01

    The present calculation, in a close-coupled approximation for the energy range up to 1,000,000 K, yields collision strengths for the electron impact excitation of S III from the ground 3p2 3P state to the excited states 3s3p3 3D0, 3P0, 3S0, 3d 3D0, 3P0, and 4s 3P0. Also obtained are those transitions' oscillator strengths, and strengths for others involving 3p2 1D and 1S. Configuration-interaction target wave functions yielding oscillator strengths that are accurate to 20 percent are used in collision strength calculations.

  5. Radiative rates for E1, E2, M1, and M2 transitions in S-like to F-like tungsten ions (W LIX to W LXVI)

    NASA Astrophysics Data System (ADS)

    Aggarwal, Kanti M.; Keenan, Francis P.

    2016-09-01

    Calculations of energy levels, radiative rates and lifetimes are reported for eight ions of tungsten, i.e. S-like (W LIX) to F-like (W LXVI). A large number of levels have been considered for each ion and extensive configuration interaction has been included among a range of configurations. For the calculations, the general-purpose relativistic atomic structure package (GRASP) has been adopted, and radiative rates (as well as oscillator strengths and line strengths) are listed for all E1, E2, M1, and M2 transitions of the ions. Comparisons have been 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).

  6. 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…

  7. 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…

  8. 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.

  9. Active site remodeling accompanies thioester bond formation in the SUMO E1

    PubMed Central

    Olsen, Shaun K.; Capili, Allan D.; Lu, Xuequan; Tan, Derek S.; Lima, Christopher D.

    2009-01-01

    E1 enzymes activate ubiquitin (Ub) and ubiquitin-like (Ubl) proteins in two steps by carboxy-terminal adenylation and thioester bond formation to a conserved catalytic cysteine in the E1 Cys domain. The structural basis for these intermediates remains unknown. Here we report crystal structures for human SUMO E1 in complex with SUMO adenylate and tetrahedral intermediate analogs at 2.45 Å and 2.6 Å, respectively. These structures show that side chain contacts to ATP·Mg are released after adenylation to facilitate a 130 degree rotation of the Cys domain during thioester bond formation that is accompanied by remodeling of key structural elements including the helix that contains the E1 catalytic cysteine, the cross-over and re-entry loops, and refolding of two helices that are required for adenylation. These changes displace side chains required for adenylation with side chains required for thioester bond formation. Mutational and biochemical analyses suggest these mechanisms are conserved in other E1s. PMID:20164921

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

    PubMed Central

    Wang, Jimin; Li, Yue; Modis, Yorgo

    2014-01-01

    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. PMID:24725935

  11. Differential induction of cytolytic susceptibility by E1A, myc, and ras oncogenes in immortalized cells.

    PubMed Central

    Cook, J L; May, D L; Wilson, B A; Walker, T A

    1989-01-01

    The E1A oncogene of adenovirus serotypes 2 and 5 induces susceptibility to the cytolytic effects of natural killer lymphocytes and activated macrophages when expressed in infected and transformed mammalian cells (cytolysis-susceptible phenotype). E1A and the oncogenes v-myc, long-terminal-repeat-promoted c-myc, and activated c-ras share the ability to immortalize transfected low-passage rodent cells. The cytolytic phenotypes of well-characterized rodent cell lines immortalized by these three oncogenes were defined. In contrast to target cells expressing the intact E1A gene, myc- and ras-expressing, immortalized primary transfectants were resistant to lysis by both types of killer cell populations. The same patterns of susceptibility (E1A) and resistance (myc and ras) to cytolysis were observed in oncogene-transfected continuous rat (REF52) and mouse (NIH 3T3) cell lines, indicating that differences in the cytolytic phenotypes associated with expression of these oncogenes are not due to cell selection during immortalization. The results suggest that the E1A oncogene may possess a functional domain that is different from those of other oncogenes, such as myc and ras, and that the activity linked to this postulated domain is dissociable from the process of immortalization. Images PMID:2526229

  12. Effect of BI-1 on insulin resistance through regulation of CYP2E1.

    PubMed

    Lee, Geum-Hwa; Oh, Kyoung-Jin; Kim, Hyung-Ryong; Han, Hye-Sook; Lee, Hwa-Young; Park, Keun-Gyu; Nam, Ki-Hoan; Koo, Seung-Hoi; Chae, Han-Jung

    2016-01-01

    Diet-induced obesity is a major contributing factor to the progression of hepatic insulin resistance. Increased free fatty acids in liver enhances endoplasmic reticulum (ER) stress and production of reactive oxygen species (ROS), both are directly responsible for dysregulation of hepatic insulin signaling. BI-1, a recently studied ER stress regulator, was examined to investigate its association with ER stress and ROS in insulin resistance models. To induce obesity and insulin resistance, BI-1 wild type and BI-1 knock-out mice were fed a high-fat diet for 8 weeks. The BI-1 knock-out mice had hyperglycemia, was associated with impaired glucose and insulin tolerance under high-fat diet conditions. Increased activity of NADPH-dependent CYP reductase-associated cytochrome p450 2E1 (CYP2E1) and exacerbation of ER stress in the livers of BI-1 knock-out mice was also observed. Conversely, stable expression of BI-1 in HepG2 hepatocytes was shown to reduce palmitate-induced ER stress and CYP2E1-dependent ROS production, resulting in the preservation of intact insulin signaling. Stable expression of CYP2E1 led to increased ROS production and dysregulation of insulin signaling in hepatic cells, mimicking palmitate-mediated hepatic insulin resistance. We propose that BI-1 protects against obesity-induced hepatic insulin resistance by regulating CYP2E1 activity and ROS production. PMID:27576594

  13. Quantitative analysis of regions of adenovirus E1A products involved in interactions with cellular proteins.

    PubMed

    Barbeau, D; Marcellus, R C; Bacchetti, S; Bayley, S T; Branton, P E

    1992-01-01

    Human adenovirus E1A proteins and oncogene products of several other DNA tumour viruses derive much of their oncogenic potential from interactions with cellular polypeptides. E1A proteins form complexes with p105Rb and a related p107 polypeptide, and with at least three other proteins (p60cycA, p130, and p300); all may be required for cell transformation. Using a series of E1A deletion mutants, we have carried out a quantitative analysis of the binding patterns of cellular proteins to E1A products. Binding of most of the proteins was affected at least partially by mutations within the amino terminal 25 residues, amino acids 36-69 within conserved region 1 (CR1), and residues 121-138 in conserved region 2 (CR2). However, the specific binding characteristics of each protein varied considerably. p300 was the only species for which binding was totally eliminated by deletions at the amino terminus. Removal of regions within CR1 eliminated binding of all species except p107 and p60cycA. Deletion of portions of CR2 reduced or eliminated binding of all proteins except p300. Thus, whereas cellular polypeptides generally were found to interact with the same three regions of E1A proteins, specific interactions varied considerably. PMID:1297336

  14. 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.

  15. Rb function is required for E1A-induced S-phase checkpoint activation.

    PubMed

    Nemajerova, A; Talos, F; Moll, U M; Petrenko, O

    2008-09-01

    It is widely accepted that adenoviral E1A exerts its influence on recipient cells through binding to the retinoblastoma (Rb) family proteins, followed by a global release of E2F factors from pocket-protein control. Our study challenges this simple paradigm by demonstrating previously unappreciated complexity. We show that E1A-expressing primary and transformed cells are characterized by the persistence of Rb-E2F1 complexes. We provide evidence that E1A causes Rb stabilization by interfering with its proteasomal degradation. Functional experiments supported by biochemical data reveal not only a dramatic increase in Rb and E2F1 protein levels in E1A-expressing cells but also demonstrate their activation throughout the cell cycle. We further show that E1A activates an Rb- and E2F1-dependent S-phase checkpoint that attenuates the growth of cells that became hyperploid through errors in mitosis and supports the fidelity DNA replication even in the absence of E2F complexes with other Rb family proteins, thereby functionally substituting for the loss of p53. Our results support the essential role of Rb and E2F1 in the regulation of genomic stability and DNA damage checkpoints.

  16. Effect of BI-1 on insulin resistance through regulation of CYP2E1

    PubMed Central

    Lee, Geum-Hwa; Oh, Kyoung-Jin; Kim, Hyung-Ryong; Han, Hye-Sook; Lee, Hwa-Young; Park, Keun-Gyu; Nam, Ki-Hoan; Koo, Seung-Hoi; Chae, Han-Jung

    2016-01-01

    Diet-induced obesity is a major contributing factor to the progression of hepatic insulin resistance. Increased free fatty acids in liver enhances endoplasmic reticulum (ER) stress and production of reactive oxygen species (ROS), both are directly responsible for dysregulation of hepatic insulin signaling. BI-1, a recently studied ER stress regulator, was examined to investigate its association with ER stress and ROS in insulin resistance models. To induce obesity and insulin resistance, BI-1 wild type and BI-1 knock-out mice were fed a high-fat diet for 8 weeks. The BI-1 knock-out mice had hyperglycemia, was associated with impaired glucose and insulin tolerance under high-fat diet conditions. Increased activity of NADPH-dependent CYP reductase-associated cytochrome p450 2E1 (CYP2E1) and exacerbation of ER stress in the livers of BI-1 knock-out mice was also observed. Conversely, stable expression of BI-1 in HepG2 hepatocytes was shown to reduce palmitate-induced ER stress and CYP2E1-dependent ROS production, resulting in the preservation of intact insulin signaling. Stable expression of CYP2E1 led to increased ROS production and dysregulation of insulin signaling in hepatic cells, mimicking palmitate-mediated hepatic insulin resistance. We propose that BI-1 protects against obesity-induced hepatic insulin resistance by regulating CYP2E1 activity and ROS production. PMID:27576594

  17. Cooperative effects for CYP2E1 differ between styrene and its metabolites

    PubMed Central

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

    2014-01-01

    Cooperative interactions are frequently observed in the metabolism of drugs and pollutants by cytochrome P450s; nevertheless, the molecular determinants for cooperativity remain elusive. Previously, we demonstrated that steady-state styrene metabolism by CYP2E1 exhibits positive cooperativity.We hypothesized that styrene metabolites have lower affinity than styrene toward CYP2E1 and limited ability to induce cooperative effects during metabolism. To test the hypothesis, we determined the potency and mechanism of inhibition for styrene and its metabolites toward oxidation of 4-nitrophenol using CYP2E1 Supersomes® and human liver microsomes.Styrene inhibited the reaction through a mixed cooperative mechanism with high affinity for the catalytic site (67 μM) and lower affinity for the cooperative site (1100 μM), while increasing substrate turnover at high concentrations. Styrene oxide and 4-vinylphenol possessed similar affinity for CYP2E1. Styrene oxide behaved cooperatively like styrene, but 4-vinylphenol decreased turnover at high concentrations. Styrene glycol was a very poor competitive inhibitor. Among all compounds, there was a positive correlation with binding and hydrophobicity.Taken together, these findings for CYP2E1 further validate contributions of cooperative mechanisms to metabolic processes, demonstrate the role of molecular structure on those mechanisms and underscore the potential for heterotropic cooperative effects between different compounds. PMID:23327532

  18. 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.

  19. 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.

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

    PubMed

    Wang, Jimin; Li, Yue; Modis, Yorgo

    2014-04-01

    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.

  1. Mitochondrial pyruvate dehydrogenase. Molecular cloning of the E1 alpha subunit and expression analysis.

    PubMed Central

    Grof, C P; Winning, B M; Scaysbrook, T P; Hill, S A; Leaver, C J

    1995-01-01

    A polymerase chain reaction-based approach was used to isolate cDNA clones encoding the E1 alpha subunit of the mitochondrial pyruvate dehydrogenase from higher plants. Putative full-length clones were identified on the basis of similarity to E1 alpha sequences from nonplant sources. Southern blot analysis revealed a small family of genes in potato (Solanum tuberosum L.), whereas in cucumber (Cucumis sativus) there are only one or two genes. Tissue-specific variation in the relative amounts of E1 alpha mRNA was observed in northern blot analysis of different potato tissues, with the highest steady-state transcript levels found in floral tissue. Measurement of pyruvate dehydrogenase activity in cucumber cotyledons showed that there is a transient increase to a maximum at 4 to 5 d postimbibition. Western blot analysis revealed that the amount of E1 alpha protein also peaks at this time. Steady-state transcript levels in germinating cucumber cotyledons also show transient accumulation, peaking 2 d postimbibition. These data are consistent with regulation of E1 alpha at the level of transcription and/or mRNA stability in postgerminative cucumber cotyledons. PMID:7659754

  2. Active site remodelling accompanies thioester bond formation in the SUMO E1

    SciTech Connect

    Olsen, Shaun K.; Capili, Allan D.; Lu, Xuequan; Tan, Derek S.; Lima, Christopher D.

    2010-03-30

    E1 enzymes activate ubiquitin (Ub) and ubiquitin-like (Ubl) proteins in two steps by carboxy-terminal adenylation and thioester bond formation to a conserved catalytic cysteine in the E1 Cys domain. The structural basis for these intermediates remains unknown. Here we report crystal structures for human SUMO E1 in complex with SUMO adenylate and tetrahedral intermediate analogues at 2.45 and 2.6 {angstrom}, respectively. These structures show that side chain contacts to ATP-Mg are released after adenylation to facilitate a 130 degree rotation of the Cys domain during thioester bond formation that is accompanied by remodelling of key structural elements including the helix that contains the E1 catalytic cysteine, the crossover and re-entry loops, and refolding of two helices that are required for adenylation. These changes displace side chains required for adenylation with side chains required for thioester bond formation. Mutational and biochemical analyses indicate these mechanisms are conserved in other E1s.

  3. Failure strength of icy lithospheres

    NASA Technical Reports Server (NTRS)

    Golombek, M. P.; Banerdt, W. B.

    1987-01-01

    Lithospheric strengths derived from friction on pre-existing fractures and ductile flow laws show that the tensile strength of intact ice under applicable conditions is actually an order of magnitude stronger than widely assumed. It is demonstrated that this strength is everywhere greater than that required to initiate frictional sliding on pre-existing fractures and faults. Because the tensile strength of intact ice increases markedly with confining pressure, it actually exceeds the frictional strength at all depths. Thus, icy lithospheres will fail by frictional slip along pre-existing fractures at yeild stresses greater than previously assumed rather than opening tensile cracks in intact ice.

  4. QM02 Strength Measurement

    SciTech Connect

    Welch, J; Wu, J.; /SLAC

    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

  5. Gaussian discriminating strength

    NASA Astrophysics Data System (ADS)

    Rigovacca, L.; Farace, A.; De Pasquale, A.; Giovannetti, V.

    2015-10-01

    We present a quantifier of nonclassical correlations for bipartite, multimode Gaussian states. It is derived from the Discriminating Strength measure, introduced for finite dimensional systems in Farace et al., [New J. Phys. 16, 073010 (2014), 10.1088/1367-2630/16/7/073010]. As the latter the new measure exploits the quantum Chernoff bound to gauge the susceptibility of the composite system with respect to local perturbations induced by unitary gates extracted from a suitable set of allowed transformations (the latter being identified by posing some general requirements). Closed expressions are provided for the case of two-mode Gaussian states obtained by squeezing or by linearly mixing via a beam splitter a factorized two-mode thermal state. For these density matrices, we study how nonclassical correlations are related with the entanglement present in the system and with its total photon number.

  6. 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.

  7. 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

  8. Regulation of the effects of CYP2E1-induced oxidative stress by JNK signaling.

    PubMed

    Schattenberg, Jörn M; Czaja, Mark J

    2014-01-01

    The generation of excessive amounts of reactive oxygen species (ROS) leads to cellular oxidative stress that underlies a variety of forms of hepatocyte injury and death including that from alcohol. Although ROS can induce cell damage through direct effects on cellular macromolecules, the injurious effects of ROS are mediated largely through changes in signal transduction pathways such as the mitogen-activated protein kinase c-Jun N-terminal kinase (JNK). In response to alcohol, hepatocytes have increased levels of the enzyme cytochrome P450 2E1 (CYP2E1) which generates an oxidant stress that promotes the development of alcoholic steatosis and liver injury. These effects are mediated in large part through overactivation of JNK that alters cell death pathways. Targeting the JNK pathway or its downstream effectors may be a useful therapeutic approach to the oxidative stress generated by CYP2E1 in alcoholic liver disease.

  9. Study of gamma-ray strength functions

    SciTech Connect

    Gardner, D.G.; Gardner, M.A.; Dietrich, F.S.

    1980-08-07

    The use of gamma-ray strength function systematics to calculate neutron capture cross sections and capture gamma-ray spectra is discussed. The ratio of the average capture width, GAMMA/sub ..gamma../-bar, to the average level spacing, D/sub obs/, both at the neutron separation energy, can be derived from such systematics with much less uncertainty than from separate systematics for values of GAMMA/sub ..gamma../-bar and D/sub obs/. In particular, the E1 gamma-ray strength function is defined in terms of the giant dipole resonance (GDR). The GDR line shape is modeled with the usual Lorentzian function and also with a new energy-dependent, Breit-Wigner (EDBW) function. This latter form is further parameterized in terms of two overlapping resonances, even for nuclei where photonuclear measurements do not resolve two peaks. In the mass ranges studied, such modeling is successful for all nuclei away from the N = 50 closed neutron shell. Near the N = 50 shell, a one-peak EDBW appears to be more appropriate. Examples of calculated neutron capture excitation functions and capture gamma-ray spectra using the EDBW form are given for target nuclei in the mass-90 region and also in the Ta-Au mass region. 20 figures.

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

    PubMed Central

    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

    2015-01-01

    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

  11. 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

  12. 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

  13. 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.

  14. 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.

  15. Genetic study of the functional organization of the colicin E1 molecule.

    PubMed Central

    Suit, J L; Fan, M L; Kayalar, C; Luria, S E

    1985-01-01

    Colicin E1 fragments obtained by genetic manipulations of the ColE1 plasmid were tested for bactericidal activity, binding to bacterial cells, and reactions with a series of anticolicin monoclonal antibodies. Two of the fragments were also tested for ability to form channels in liposomal vesicles. The results are in agreement with studies from chemically and enzymatically derived colicin fragments, assigning the receptor binding activity to the central part of the molecule and the killing activity to a region near the carboxyl terminus. PMID:2579061

  16. Folded state of the integral membrane colicin E1 immunity protein in solvents of mixed polarity.

    PubMed

    Taylor, R M; Zakharov, S D; Bernard Heymann, J; Girvin, M E; Cramer, W A

    2000-10-10

    The colicin E1 immunity protein (ImmE1), a 13.2-kDa hydrophobic integral membrane protein localized in the Escherichia coli cytoplasmic membrane, protects the cell from the lethal, channel-forming activity of the bacteriocin, colicin E1. Utilizing its solubility in organic solvents, ImmE1 was purified by 1-butanol extraction of isolated membranes, followed by gel filtration and ion-exchange chromatography in a chloroform/methanol/H(2)O (4:4:1) solvent system. Circular dichroism analysis indicated that the alpha-helical content of ImmE1 is approximately 80% in 1-butanol or 2,2,2-trifluoroethanol, consistent with a previous membrane-folding model with three extended hydrophobic transmembrane helical domains, H1-H3. Each of these extended hydrophobic domains contains a centrally located single Cys residue that could be used as a probe of protein structure. The presence of tertiary structure of purified ImmE1 in a solvent of mixed polarity, chloroform/methanol/H(2)O (4:4:1) was demonstrated by (i) the constraints on Tyr residues shown by the amplitude of near-UV circular dichroism spectra in the wavelength interval, 270-285 nm; (ii) the correlation between the near-UV Tyr CD spectrum of single and double Cys-to-X mutants of the Imm protein and their in vivo activity; (iii) the upfield shift of methyl groups in a 1D NMR spectrum, a 2D- HSQC NMR spectrum of ImmE1 in the mixed polarity solvent mixture, and a broadening and disappearance of the indole (1)H proton resonance from Trp94 in H3 by a spin label attached to Cys16 in the H2 hydrophobic domain; (iv) near-UV circular dichroism spectra with a prominent ellipticity band centered at 290 nm from a single Trp inserted into the extended hydrophobic domains. It was concluded that the colicin E1 immunity protein adopts a folded conformation in chloroform/methanol/H(2)O (4:4:1) that is stabilized by helix-helix interactions. Analysis of the probable membrane folding topology indicated that several Tyr residues in the bilayer

  17. A bilateral antidiuresis to renal artery infusion of prostaglandin E1 in dogs treated with phenylbutazone

    PubMed Central

    Hall, W. J.; Hensey, O. J.; O'Neill, P.; Sheehan, J. D.

    1978-01-01

    1. In acute experiments, high levels of endogenous prostaglandins, provoked by operative stress, could obscure or alter the actions of infused prostaglandins on the kidney. For this reason we decided to compare the effects of infusing prostaglandin E1 into the renal artery of the dog before and after the administration of phenylbutazone, a prostaglandin synthetase inhibitor. 2. Infusion of prostaglandin E1 into the left renal artery of the pre-phenylbutazone treated dog undergoing a mannitol diuresis increased renal plasma flow, glomerular filtration rate and the excretion of salt and water. The findings are in general agreement with those reported by others. 3. Following phenylbutazone administration the vascular and saluretic actions of prostaglandin E1 were unchanged but a reduced diuretic effect was observed. The response to a low dose of prostaglandin E1 (0·05 μg/min) was reduced from 1·46 ± 0·15 to 0·96 ± 0·16 ml./min (P < 0·001) and the response to a high dose (0·5 μg/min) from 1·82 ± 0·19 to 0·99 ± 0·31 ml./min (P < 0·002). 4. A significantly less dilute urine was excreted during prostaglandin infusion in the dog after phenylbutazone treatment than before. The reduction in the diuretic response was of the same order as the decrease in the free water clearance response, while the increase in osmolar clearance was unchanged. 5. In water-loaded dogs treated with phenylbutazone, infusion of prostaglandin E1 into the left renal artery had a biphasic effect on urine output from the left kidney. An initial diuretic response to a low dose of prostaglandin E1 disappeared with the infusion of higher doses, and antidiuresis developed in the immediate post-infusion period. 6. As prostaglandin was infused into the left kidney progressive antidiuresis was seen in the non-infused right kidney. 7. It is concluded that endogenous prostaglandins do not obscure or alter the vascular and saluretic actions of intrarenal prostaglandin E1. The findings question

  18. Sound power measurements on the M1E1 engine and total power pack

    NASA Astrophysics Data System (ADS)

    Schomer, P. D.

    1983-05-01

    A new technique to measure power was used to gather one-third octave emissions data on the M1E1 engine (AGT 1500) at the Stratford Army Engine Plant, CT and on the M1E1 power pack at Aberdeen Proving Ground, MD. To calculate sound power, acoustic intensity (the product of the scaler pressure and the vector velocity) was measured and integrated over an arbitrary surface enclosing the source. The data gathered and test results show this new technique is very powerful and robust and is applicable to many machinery quieting, source emission, or sound transmission measurements.

  19. 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,...

  20. Association between CYP2E1 polymorphisms and risk of gastric cancer: An updated meta-analysis of 32 case-control studies

    PubMed Central

    ZHANG, MING-XING; LIU, KAI; WANG, FU-GANG; WEN, XIAO-WEN; SONG, XI-LIN

    2016-01-01

    Previous studies suggested that RsaI/PstI and DraI polymorphisms on cytochrome P450 2E1 (CYP2E1) may be associated with susceptibility to gastric cancer (GC). However, this association remains ambiguous. A meta-analysis of previously published studies was performed in an attempt to elucidate this association. The odds ratio and 95% confidence interval were used to assess the strength of the association. In the overall analyses of RsaI/PstI and DraI, no association was identified. In the subgroup analyses, RsaI/PstI was identified to increase the risk of GC in the smoking population. In addition, in the previous studies of interactions with other genes, RsaI/PstI was revealed to be associated with increased GC risks when glutathione S-transferase-µ-1 or glutathione S-transferase θ-1 was null or DraI was homozygous wild-type. However, these stratified analyses were lacking credibility due to the limitation of correlational study numbers. In conclusion, CYP2E1 polymorphisms revealed no association with the risk of GC. PMID:27284439

  1. 26 CFR 1.6050E-1 - Reporting of State and local income tax refunds.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 13 2011-04-01 2011-04-01 false Reporting of State and local income tax refunds... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Information Returns § 1.6050E-1 Reporting of State and local income tax refunds. (a) Applicability. Section 6050E and this section apply to...

  2. 26 CFR 1.6050E-1 - Reporting of State and local income tax refunds.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 13 2010-04-01 2010-04-01 false Reporting of State and local income tax refunds... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES Information Returns § 1.6050E-1 Reporting of State and local income tax refunds. (a) Applicability. Section 6050E and this section apply to any refund officer...

  3. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who... 40 Protection of Environment 6 2013-07-01 2013-07-01 false Designation Testing Checklist E Figure E-1 to Subpart E of Part 53 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

  4. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who... 40 Protection of Environment 6 2012-07-01 2012-07-01 false Designation Testing Checklist E Figure E-1 to Subpart E of Part 53 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

  5. 40 CFR Figure E-1 to Subpart E of... - Designation Testing Checklist

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 CFR Part 53 or 40 CFR Part 50, Appendix L Verification Comments (Includes documentation of who... 40 Protection of Environment 6 2014-07-01 2014-07-01 false Designation Testing Checklist E Figure E-1 to Subpart E of Part 53 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY...

  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. 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...

  8. 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...

  9. 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...

  10. 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...

  11. 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...

  12. 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.

  13. VirE1-Mediated Resistance to Crown Gall in Transgenic Arabidopsis thaliana.

    PubMed

    Humann, Jodi; Andrews, Sarah; Ream, Walt

    2006-01-01

    ABSTRACT Crown gall disease, caused by Agrobacterium tumefaciens, remains a serious agricultural problem despite current biocontrol methods. Agrobacterium tumefaciens transfers single-stranded DNA (T-strands) into plant cells along with several virulence proteins, including a single-stranded DNA-binding protein (VirE2). In plant cells, T-strands are protected from nucleases and targeted to the nucleus by VirE2, which is essential for efficient transmission (transfer and integration) of T-strands. VirE1 is the secretory chaperone for VirE2; it prevents VirE2 from forming aggregates and from binding the T-strands in bacterial cells. Therefore, we hypothesized that sufficient quantities of VirE1 expressed in plant cells might block T-DNA transmission by preventing VirE2 from binding T-strands. Here we show that root explants from Arabidopsis thaliana plants that expressed virE1 formed 3.5-fold fewer tumors than roots from plants without virE1. Also, this resistance was specific for VirE2-mediated Agrobacterium transformation. Plants that have been genetically altered to resist crown gall may prove more effective than biological control. PMID:18944210

  14. 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...

  15. 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. ABC does not have more than 100 partners, and partnership A is entitled to notice under section...

  16. Phyllanthus urinaria extract attenuates acetaminophen induced hepatotoxicity: involvement of cytochrome P450 CYP2E1.

    PubMed

    Hau, Desmond Kwok Po; Gambari, Roberto; Wong, Raymond Siu Ming; Yuen, Marcus Chun Wah; Cheng, Gregory Yin Ming; Tong, Cindy Sze Wai; Zhu, Guo Yuan; Leung, Alexander Kai Man; Lai, Paul Bo San; Lau, Fung Yi; Chan, Andrew Kit Wah; Wong, Wai Yeung; Kok, Stanton Hon Lung; Cheng, Chor Hing; Kan, Chi Wai; Chan, Albert Sun Chi; Chui, Chung Hin; Tang, Johnny Cheuk On; Fong, David Wang Fun

    2009-08-01

    Acetaminophen is a commonly used drug for the treatment of patients with common cold and influenza. However, an overdose of acetaminophen may be fatal. In this study we investigated whether mice, administered intraperitoneally with a lethal dose of acetaminophen, when followed by oral administration of Phyllanthus urinaria extract, may be prevented from death. Histopathological analysis of mouse liver sections showed that Phyllanthus urinaria extract may protect the hepatocytes from acetaminophen-induced necrosis. Therapeutic dose of Phyllanthus urinaria extract did not show any toxicological phenomenon on mice. Immunohistochemical staining with the cytochrome P450 CYP2E1 antibody revealed that Phyllanthus urinaria extract reduced the cytochrome P450 CYP2E1 protein level in mice pre-treated with a lethal dose of acetaminophen. Phyllanthus urinaria extract also inhibited the cytochrome P450 CYP2E1 enzymatic activity in vitro. Heavy metals, including arsenic, cadmium, mercury and lead, as well as herbicide residues were not found above their detection limits. High performance liquid chromatography identified corilagin and gallic acid as the major components of the Phyllanthus urinaria extract. We conclude that Phyllanthus urinaria extract is effective in attenuating the acetaminophen induced hepatotoxicity, and inhibition of cytochrome P450 CYP2E1 enzyme may be an important factor for its therapeutic mechanism.

  17. Ubiquitination Accomplished: E1 and E2 Enzymes Were Not Necessary.

    PubMed

    Nakasone, Mark A; Huang, Danny T

    2016-06-16

    Qiu et al. (2016) show that a mono-ADP-ribosyltransferase, SdeA, from Legionella pneumophila catalyzes ADP-ribosylation of ubiquitin, allowing SdeA to modify substrate with ubiquitin in the absence of E1 and E2 enzymes. PMID:27315555

  18. Isolation and characterization of mutants affecting functional domains of ColE1 RNAI.

    PubMed

    Dooley, T P; Tamm, J; Polisky, B

    1985-11-01

    The control of DNA replication initiation in the plasmid ColE1 is mediated by RNAI, a 108 nucleotide plasmid-encoded RNA that is entirely complementary to the 5'-terminal region of the replication primer RNA. RNAI acts in trans to inhibit primer maturation. Previously, we constructed a plasmid in which the ColE1 RNAI was separated from the primer and placed under transcriptional control of the Serratia marcesens tryptophan promoter. This plasmid provides RNAI in trans in vivo and mediates ColE1-type incompatibility. To determine the critical structural and functional domains of RNAI, we have undertaken a mutational analysis of the RNAI gene carried by this plasmid. We have selected mutants that no longer mediate ColE1-type incompatibility in trans. From the DNA sequences of 18 mutants we have identified mutations at nine new sites in RNAI. In addition, we have determined the secondary structural features of several mutant RNAI species and compared them to wild-type RNAI. Analysis of these mutations has revealed several key features of RNAI secondary structure and function. The domains of RNAI identified in this work which are essential for its function are: the single-stranded loop regions; the integrity of the double-stranded stems; and the single-stranded 5' terminus.

  19. 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.

  20. 26 CFR 1.6050E-1 - Reporting of State and local income tax refunds.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 13 2012-04-01 2012-04-01 false Reporting of State and local income tax refunds... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Information Returns § 1.6050E-1 Reporting of State and local income tax refunds. (a) Applicability. Section 6050E and this section apply to...

  1. 26 CFR 1.6050E-1 - Reporting of State and local income tax refunds.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 13 2014-04-01 2014-04-01 false Reporting of State and local income tax refunds... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Information Returns § 1.6050E-1 Reporting of State and local income tax refunds. (a) Applicability. Section 6050E and this section apply to...

  2. 26 CFR 1.6050E-1 - Reporting of State and local income tax refunds.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 13 2013-04-01 2013-04-01 false Reporting of State and local income tax refunds... (CONTINUED) INCOME TAX (CONTINUED) INCOME TAXES (CONTINUED) Information Returns § 1.6050E-1 Reporting of State and local income tax refunds. (a) Applicability. Section 6050E and this section apply to...

  3. 26 CFR 1.1397E-1 - Qualified zone academy bonds.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 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...

  4. 42 CFR 52e.1 - To what programs do these regulations apply?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 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 at... HEART, LUNG, AND BLOOD INSTITUTE GRANTS FOR PREVENTION AND CONTROL PROJECTS § 52e.1 To what programs...

  5. 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...

  6. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 8 2010-04-01 2010-04-01 false Installment obligations transmitted at death....691(e)-1 Installment obligations transmitted at death when prior law applied. (a) In general—(1... death of a holder of such obligations were required to be reported in the return of the decedent for...

  7. 26 CFR 1.691(e)-1 - Installment obligations transmitted at death when prior law applied.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 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 general... obligations at the death of a holder of such obligations were required to be reported in the return of...

  8. 26 CFR 1.1244(e)-1 - Records to be kept.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 11 2011-04-01 2011-04-01 false Records to be kept. 1.1244(e)-1 Section 1.1244... Records to be kept. (a) By the corporation—(1) Mandatory records. A plan to issue pre-November 1978 stock... records to be maintained. In addition, a person who owns section 1244 stock in a corporation...

  9. 26 CFR 1.1244(e)-1 - Records to be kept.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 11 2014-04-01 2014-04-01 false Records to be kept. 1.1244(e)-1 Section 1.1244... Records to be kept. (a) By the corporation—(1) Mandatory records. A plan to issue pre-November 1978 stock... records to be maintained. In addition, a person who owns section 1244 stock in a corporation...

  10. 26 CFR 1.1244(e)-1 - Records to be kept.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 11 2013-04-01 2013-04-01 false Records to be kept. 1.1244(e)-1 Section 1.1244... Records to be kept. (a) By the corporation—(1) Mandatory records. A plan to issue pre-November 1978 stock... records to be maintained. In addition, a person who owns section 1244 stock in a corporation...

  11. 26 CFR 1.1244(e)-1 - Records to be kept.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 11 2012-04-01 2012-04-01 false Records to be kept. 1.1244(e)-1 Section 1.1244... Records to be kept. (a) By the corporation—(1) Mandatory records. A plan to issue pre-November 1978 stock... records to be maintained. In addition, a person who owns section 1244 stock in a corporation...

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

    PubMed

    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; Cole, Stewart T

    2015-08-01

    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.

  13. 78 FR 15597 - Special Conditions: GE Aviation CT7-2E1 Turboshaft Engine Model

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-03-12

    ... conditions are issued for the General Electric Aviation (GE) CT7-2E1 engine model. This engine model will...;having general applicability and legal effect, most of which are keyed #0;to and codified in the Code of... published on July 20, 2012 (77 FR 42677). We received six comments from European Aviation Safety...

  14. 11 CFR 101.1 - Candidate designations (2 U.S.C. 432(e)(1)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... accordance with 11 CFR 102.12. A candidate shall designate his or her principal campaign committee by filing... specified at 11 CFR part 105). Each principal campaign committee shall register, designate a depository, and... DESIGNATIONS (2 U.S.C. 432(e)) § 101.1 Candidate designations (2 U.S.C. 432(e)(1)). (a) Principal...

  15. 11 CFR 300.60 - Scope (2 U.S.C. 441i(e)(1)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 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 300.60 Federal Elections FEDERAL ELECTION COMMISSION BIPARTISAN CAMPAIGN REFORM ACT OF...

  16. 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....

  17. 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....

  18. 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....

  19. 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....

  20. 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

  1. 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.

  2. 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.

  3. Subregions of the adenovirus E1A transactivation domain target multiple components of the TFIID complex.

    PubMed Central

    Geisberg, J V; Chen, J L; Ricciardi, R P

    1995-01-01

    Transcriptional activation by the adenovirus E1A 289R protein requires direct contacts with the TATA box-binding protein (TBP) and also displays a critical requirement for TBP-associated factors (TAFs) (T.G. Boyer and A. J. Berk, Genes Dev. 7:1810-1823, 1993; J. V. Geisberg, W. S. Lee, A. J. Berk, and R. P. Ricciardi, Proc. Natl. Acad. Sci. USA 91:2488-2492, 1994; W. S. Lee, C. C. Kao, G. O. Bryant, X. Liu, and A. J. Berk, Cell 67:365-376, 1991; and Q. Zhou, P. M. Lieberman, T. G. Boyer, and A. J. Berk, Genes Dev. 6:1964-1974, 1992). In this report, we demonstrate that the activation domain of E1A (CR3) specifically binds to two TAFs, human TAFII250 (hTAFII250) and Drosophila TAFII110 (dTAFII110). These interactions can take place both in vivo and in vitro and require the carboxy-terminal region of CR3; the zinc finger region of CR3, which binds TBP, is not needed to bind these TAFs. We mapped the E1A-binding sites on hTAFII250 to an internal region that contains a number of structural motifs, including an HMG box, a bromodomain, and direct repeats. This represents the first demonstration that hTAFII250 may serve as a target of a transcriptional activator. We also mapped the E1A binding on dTAFII110 to its C-terminal region. This is of significance since, by contrast, Sp1-mediated activation requires binding to the N-terminal domain of dTAFII110. Thus, distinct surfaces of dTAFII110 can serve as target sites for different activators. Our results indicate that E1A may activate transcription, in part, through direct contacts of the CR3 subdomains with selected components of the TFIID complex. PMID:7565781

  4. 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

  5. Antibody directed to a synthetic peptide encoding the NH2-terminal 16 amino acids of the adenovirus type 2 E1B-53K tumor antigen recognizes the E1B-20K tumor antigen.

    PubMed

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

    1984-01-15

    A peptide, H2N-Glu-Arg-Arg-Asn-Pro-Ser-Glu-Arg-Gly-Val-Pro-Ala-Gly-Phe-Ser-Gly-(Cys )COOH, containing the amino acid sequence at the NH2 terminus of the adenovirus type 2 (Ad2) E1B-coded large T antigen (E1B-53K) has been synthesized. Anti-peptide antibody was generated in rabbits and used to immunoprecipitate Ad T antigens from Ad2 early infected cell extracts. In addition to the expected E1B-53K T antigen, anti-peptide antibody precipitated the Ad2 E1B-20K T antigen that was previously shown to be related to E1B-53K (M. Green, K.H. Brackmann, M.A. Cartas, and T. Matsuo, J. Virol. 42, 30-41, 1982). Anti-peptide prepared against the COOH terminus of the E1B-53K T antigen or against the NH2 terminus of the E1B-19K T antigen did not precipitate the E1B-20K T antigen. These data suggest that the Ad2 E1B-20K T antigen initiates translation at nucleotide 2016 in reading frame 3, as does E1B-53K. The viral mRNA that encodes the E1B-20K T antigen has not been identified.

  6. Tensile strength of restorative resins.

    PubMed

    Zidan, O; Asmussen, E; Jørgensen, K D

    1980-06-01

    The purpose of the present work was to measure the tensile strength of restorative resins and to study the effect of the method of measurement on the recorded results. A direct pull method using dumb-bell shaped specimens was used. The tensile strength of the resins was also tested using the diametral compression method suggested by the A.D.A. It was found that the method of testing affects the results. Although the diametral compression method is a simple method, it cannot be considered reliable for all types of material. The tensile strength of the conventional composites was significantly higher than the tensile strength of the microfilled composites.

  7. Physiological Effects of Strength Training and Various Strength Training Devices.

    ERIC Educational Resources Information Center

    Wilmore, Jack H.

    Current knowledge in the area of muscle physiology is a basis for a discussion on strength training programs. It is now recognized that the expression of strength is related to, but not dependent upon, the size of the muscle and is probably more related to the ability to recruit more muscle fibers in the contraction, or to better synchronize their…

  8. Intestinal CYP2E1: A mediator of alcohol-induced gut leakiness

    PubMed Central

    Forsyth, Christopher B.; Voigt, Robin M.; Keshavarzian, Ali.

    2014-01-01

    Chronic alcohol use can result in many pathological effects including alcoholic liver disease (ALD). While alcohol is necessary for the development of ALD, only 20–30% of alcoholics develop alcoholic steatohepatitis (ASH) with progressive liver disease leading to cirrhosis and liver failure (ALD). This suggests that while chronic alcohol consumption is necessary it is not sufficient to induce clinically relevant liver damage in the absence of a secondary risk factor. Studies in rodent models and alcoholic patients show that increased intestinal permeability to microbial products like endotoxin play a critical role in promoting liver inflammation in ALD pathogenesis. Therefore identifying mechanisms of alcohol-induced intestinal permeability is important in identifying mechanisms of ALD and for designing new avenues for therapy. Cyp2e1 is a cytochrome P450 enzyme that metabolizes alcohol has been shown to be upregulated by chronic alcohol use and to be a major source of oxidative stress and liver injury in alcoholics and in animal and in vitro models of chronic alcohol use. Because Cyp2e1 is also expressed in the intestine and is upregulated by chronic alcohol use, we hypothesized it could play a role in alcohol-induced intestinal hyperpermeability. Our in vitro studies with intestinal Caco-2 cells and in mice fed alcohol showed that circadian clock proteins CLOCK and PER2 are required for alcohol-induced permeability. We also showed that alcohol increases Cyp2e1 protein and activity but not mRNA in Caco-2 cells and that an inhibitor of oxidative stress or siRNA knockdown of Cyp2e1 prevents the increase in CLOCK or PER2 proteins and prevents alcohol-induced hyperpermeability. With our collaborators we have also shown that Cyp2e1 knockout mice are resistant to alcohol-induced gut leakiness and liver inflammation. Taken together our data support a novel Cyp2e1-circadian clock protein mechanism for alcohol-induced gut leakiness that could provide new avenues for

  9. Intestinal CYP2E1: A mediator of alcohol-induced gut leakiness.

    PubMed

    Forsyth, Christopher B; Voigt, Robin M; Keshavarzian, Ali

    2014-01-01

    Chronic alcohol use can result in many pathological effects including alcoholic liver disease (ALD). While alcohol is necessary for the development of ALD, only 20-30% of alcoholics develop alcoholic steatohepatitis (ASH) with progressive liver disease leading to cirrhosis and liver failure (ALD). This suggests that while chronic alcohol consumption is necessary it is not sufficient to induce clinically relevant liver damage in the absence of a secondary risk factor. Studies in rodent models and alcoholic patients show that increased intestinal permeability to microbial products like endotoxin play a critical role in promoting liver inflammation in ALD pathogenesis. Therefore identifying mechanisms of alcohol-induced intestinal permeability is important in identifying mechanisms of ALD and for designing new avenues for therapy. Cyp2e1 is a cytochrome P450 enzyme that metabolizes alcohol has been shown to be upregulated by chronic alcohol use and to be a major source of oxidative stress and liver injury in alcoholics and in animal and in vitro models of chronic alcohol use. Because Cyp2e1 is also expressed in the intestine and is upregulated by chronic alcohol use, we hypothesized it could play a role in alcohol-induced intestinal hyperpermeability. Our in vitro studies with intestinal Caco-2 cells and in mice fed alcohol showed that circadian clock proteins CLOCK and PER2 are required for alcohol-induced permeability. We also showed that alcohol increases Cyp2e1 protein and activity but not mRNA in Caco-2 cells and that an inhibitor of oxidative stress or siRNA knockdown of Cyp2e1 prevents the increase in CLOCK or PER2 proteins and prevents alcohol-induced hyperpermeability. With our collaborators we have also shown that Cyp2e1 knockout mice are resistant to alcohol-induced gut leakiness and liver inflammation. Taken together our data support a novel Cyp2e1-circadian clock protein mechanism for alcohol-induced gut leakiness that could provide new avenues for

  10. Activities of virE1 and the VirE1 secretion chaperone in export of the multifunctional VirE2 effector via an Agrobacterium type IV secretion pathway.

    PubMed

    Zhao, Z; Sagulenko, E; Ding, Z; Christie, P J

    2001-07-01

    Agrobacterium tumefaciens uses a type IV secretion system to deliver oncogenic nucleoprotein particles and effector proteins, such as the multifunctional VirE2 protein, to plant cells. In this study, we examined the function of virE1 and its product, the VirE1 secretion chaperone, in mediating VirE2 export. A nonpolar virE1 null mutant accumulated low levels of VirE2, and trans expression of virE1 in this mutant only partially restored VirE2 abundance. Deletion of virE1 did not affect transcription but decreased translation of virE2, as shown by analysis of lacZ transcriptional and translational fusions. VirE2 was stable for a prolonged period, more than 6 h, when it was expressed in cis with virE1, and it exhibited half-lives of about 2 h when it was expressed in trans with virE1 and less than 10 min when it was expressed in the absence of virE1, as shown by pulse-chase experiments. VirE1 stabilized VirE2 via an interaction with a domain near the N terminus of VirE2, as shown by analyses of VirE2 truncation and insertion mutants synthesized in A. tumefaciens. VirE1 self-association was demonstrated by using bacteriophage lambda cI repressor fusion and pull-down assays, and evidence of VirE1 homomultimerization in vivo was obtained by native polyacrylamide gel electrophoresis and gel filtration chromatography. A putative VirE1-VirE2 complex with a molecular mass of about 70 to 80 kDa was detected by gel filtration chromatography of extracts from wild-type cells, whereas higher-order VirE2 complexes or aggregates were detected in extracts from a virE1 mutant. Taken together, our findings show that virE1 contributes in several ways to VirE2 export:(i) virE1 regulates efficient virE2 translation in the context of expression from the native P(virE) promoter; (ii) the VirE1 secretion chaperone stabilizes VirE2, most probably via an interaction with an N-terminal domain; and (iii) VirE1 forms a VirE1-VirE2 complex with a predicted 2:1 stoichiometry that inhibits assembly

  11. 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

  12. 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…

  13. Small Signal Modeling and Control of the Hydrogen Mixer for Facility E1

    NASA Technical Reports Server (NTRS)

    Barbieri, Enrique

    2001-01-01

    We have undertaken the theoretical modelling of an existing liquid hydrogen (LH2) and gas hydrogen (GH2) mixer subsystem of the E1 Ground Test Facility at NASA John C. Stennis Space Center. The E1 test facility carries out comprehensive ground-based testing and certification of various liquid rocket engines and their components. The mixer described in this work is responsible for combining high pressure before it is fed into a test article. The desired properties are maintained by precise control of the mixture of LH2 and GH2 flows. The mixer is modelled as a general multi-flow lumped volume for single constituent fluids using density and internal energy as states. The set of nonlinear differential equations is linearized about an equilibrium point and the resulting two-state, 3-input linear model is analyzed as a possible candidate for control design.

  14. Small Signal Modelling and Control of the Hydrogen Mixer for Facility E1

    NASA Technical Reports Server (NTRS)

    Barbieri, Enrique

    2003-01-01

    We have undertaken the theoretical modelling of an existing liquid hydrogen (LH2) and gas hydrogen (GH2) mixer subsystem of the E1 Ground Test Facility at NASA John C. Stennis Space Center. The E1 test facility carries out comprehensive ground-based testing and certification of various liquid rocket engines and their components. The mixer described in this work is responsible for combining high pressure LH2 and GH2 to produce a hydrogen flow that meets certain thermodynamic properties before it is fed into a test article. The desired properties are maintained by precise control of the mixture of LH2 and GH2 flows. The mixer is modelled as a general multi-flow lumped volume for single constituent fluids using density and internal energy as states. The set of nonlinear differential equations is linearized about an equilibrium point and the resulting two-state, 3-input linear model is analyzed as a possible candidate for control design.

  15. [Vasodilation effects of prostaglandin E1 in patients with precapillary pulmonary hypertension].

    PubMed

    Munclinger, M; Kautzner, J; Serf, B

    1990-04-01

    Prostaglandin E1. (Prostavasin, Schwarz, FRG) was administered in a short-term infusion, 5-30 ng/kg/min., to five patients with precapillary pulmonary hypertension associated with lung disease. Significant pulmonary vasodilatation characterized by a drop of the median pressure in the pulmonary artery by 15% and of the pulmonary vascular resistance by 31% was achieved only in two patients. The authors observed a minimal incidence of side-effects; systemic hypotension observed in three patients was an undesirable manifestaútion. Prostaglandin E1 extends possibilities of a vasodilatating influence on precapillary pulmonary hypertension in lung disease. With regard to the different individual reactivity of patients it should be administered in this indication only under conditions of haemodynamic monitoring.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. Mouse spermatocytes express CYP2E1 and respond to acrylamide exposure.

    PubMed

    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.

  2. Strength properties of fly ash based controlled low strength materials.

    PubMed

    Türkel, S

    2007-08-25

    Controlled low strength material (CLSM) is a flowable mixture that can be used as a backfill material in place of compacted soils. Flowable fill requires no tamping or compaction to achieve its strength and typically has a load carrying capacity much higher than compacted soils, but it can still be excavated easily. The selection of CLSM type should be based on technical and economical considerations for specific applications. In this study, a mixture of high volume fly ash (FA), crushed limestone powder (filler) and a low percentage of pozzolana cement have been tried in different compositions. The amount of pozzolana cement was kept constant for all mixes as, 5% of fly ash weight. The amount of mixing water was chosen in order to provide optimum pumpability by determining the spreading ratio of CLSM mixtures using flow table method. The shear strength of the material is a measure of the materials ability to support imposed stresses on the material. The shear strength properties of CLSM mixtures have been investigated by a series of laboratory tests. The direct shear test procedure was applied for determining the strength parameters Phi (angle of shearing resistance) and C(h) (cohesion intercept) of the material. The test results indicated that CLSM mixtures have superior shear strength properties compared to compacted soils. Shear strength, cohesion intercept and angle of shearing resistance values of CLSM mixtures exceeded conventional soil materials' similar properties at 7 days. These parameters proved that CLSM mixtures are suitable materials for backfill applications.

  3. Purification, crystallization and preliminary X-ray diffraction analysis of human enolase-phosphatase E1

    SciTech Connect

    Wang, Hui; Pang, Hai; Ding, Yi; Li, Yi; Wu, Xiao’ai; Rao, Zihe

    2005-05-01

    Recombinant human E1 enzyme has been crystallized using the hanging-drop vapour-diffusion method and diffraction-quality crystals were grown at 291 K using PEG 4000 as precipitant. Enolase-phosphatase E1 (MASA) is a bifunctional enzyme in the ubiquitous methionine-salvage pathway and catalyzes the continuous reaction of 2,3-diketo-5-methylthio-1-phosphopentane to yield the acireductone metabolite. Recombinant human E1 enzyme has been crystallized using the hanging-drop vapour-diffusion method and diffraction-quality crystals were grown at 291 K using PEG 4000 as precipitant. Diffraction data were collected to 1.7 Å resolution from SeMet-derivative crystals at 100 K using synchrotron radiation. The crystals belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 54.02, b = 57.55, c = 87.32 Å. The structure was subsequently solved by the multi-wavelength anomalous diffraction (MAD) phasing method.

  4. Evidence for selective regulation of the phosphorylation of myocyte proteins by isoproterenol and prostaglandin E1.

    PubMed

    Hayes, J S; Bowling, N; King, K L; Boder, G B

    1982-01-12

    Both isoproterenol and prostaglandin E1 increased the activation state of cyclic AMP-dependent protein kinase in cultured myocytes; however, only isoproterenol enhanced phosphorylase activity and contractile state. Following the incubation of intact myocytes with 32PO3-(4), 32 phosphoproteins were resolved from total cellular proteins by electrophoresis in sodium dodecyl sulfate polyacrylamide gels followed by autoradiography. Isoproterenol stimulated 32PO3-(4) incorporation into 16 proteins, including 2 phosphoproteins not observed under control conditions. By contrast, prostaglandin E1 neither caused a measurable change in the protein phosphorylation pattern nor interfered with isoproterenol's capacity to do so. Isoproterenol stimulated myocyte protein phosphorylation in either the presence or absence of extracellular Ca2+. The results suggest that the regulation of protein phosphorylation following adenylate cyclase stimulation is: (1) an agonist-specific process and not due solely to a random accumulation of intracellular cycle AMP and activation of protein kinase; (2) the Ca2+ mobilization component of beta-receptor activation does not account for the paradoxical effects of isoproterenol and prostaglandin E1; (3) activation of cyclic AMP-dependent protein kinase does not always result in an enhancement of protein phosphorylation.

  5. Induction of E1A-responsive negative factors for transcription of the fibronectin gene in adenovirus E1-transformed rat cells.

    PubMed Central

    Nakamura, T; Nakajima, T; Tsunoda, S; Nakada, S; Oda, K; Tsurui, H; Wada, A

    1992-01-01

    The level of fibronectin (FN) gene expression is very high in resting rat 3Y1 cells but greatly decreased in adenovirus E1-transformed cells. To study the mechanism of this down-regulation, nuclear factors binding to the 5'-flanking region of the FN gene were analyzed by gel retardation assay and DNase I footprinting. Nuclear factors that were present in the transformed cells but nearly absent in resting 3Y1 cells interacted with multiple sites of the promoter region. Oligonucleotide competition with the FN promoter-chloramphenicol acetyltransferase (CAT) reporter constructs (pFCAT) for these factors in the transformed cells indicated that all of them had a negative effect on FN gene expression. Of them, a factor(s) (G10BP) binding to the G10 stretch from positions -239 to -230 and to two GC boxes consisting of the G10 stretch with one internal C residue insertion from positions -105 to -95 and -54 to -44 had the strongest repressive activity. Introduction of substitutive mutations into these G-rich sequences resulted in the increase in CAT activity of pFCAT in the transformed cells. The recognition sequences of G10BP and Sp1 overlap in two GC boxes. G10BP has stronger affinity for heparin and GC boxes than does Sp1, suggesting that G10BP may repress FN gene transcription by displacing Sp1. Images PMID:1404598

  6. 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

  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. Effects of ethanol on CYP2E1 levels and related oxidative stress using a standard balanced diet.

    PubMed

    Azzalis, Ligia A; Fonseca, Fernando L A; Simon, Karin A; Schindler, Fernanda; Giavarotti, Leandro; Monteiro, Hugo P; Videla, Luis A; Junqueira, Virgínia B C

    2012-07-01

    Expression of cytochrome P4502E1 (CYP2E1) is very much influenced by nutritional factors, especially carbohydrate consumption, and various results concerning the expression of CYP2E1 were obtained with a low-carbohydrate diet. This study describes the effects of ethanol treatment on CYP2E1 levels and its relationship with oxidative stress using a balanced standard diet to avoid low or high carbohydrate consumption. Rats were fed for 1, 2, 3, or 4 weeks a commercial diet plus an ethanol-sucrose solution. The results have shown that ethanol administration was associated with CYP2E1 induction and stabilization without related oxidative stress. Our findings suggest that experimental models with a low-carbohydrate/high-fat diet produce some undesirable CYP2E1 changes that are not present when a balanced standard diet is given.

  9. 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

  10. Brucella abortus ΔrpoE1 confers protective immunity against wild type challenge in a mouse model of brucellosis.

    PubMed

    Willett, Jonathan W; Herrou, Julien; Czyż, Daniel M; Cheng, Jason X; Crosson, Sean

    2016-09-30

    The Brucella abortus general stress response (GSR) system regulates activity of the alternative sigma factor, σ(E1), which controls transcription of approximately 100 genes and is required for persistence in a BALB/c mouse chronic infection model. We evaluated the host response to infection by a B. abortus strain lacking σ(E1) (ΔrpoE1), and identified pathological and immunological features that distinguish ΔrpoE1-infected mice from wild-type (WT), and that correspond with clearance of ΔrpoE1 from the host. ΔrpoE1 infection was indistinguishable from WT in terms of splenic bacterial burden, inflammation and histopathology up to 6weeks post-infection. However, Brucella-specific serum IgG levels in ΔrpoE1-infected mice were 5 times higher than WT by 4weeks post-infection, and remained significantly higher throughout the course of a 12-week infection. Total IgG and Brucella-specific IgG levels peaked strongly in ΔrpoE1-infected mice at 6weeks, which correlated with reduced splenomegaly and bacterial burden relative to WT-infected mice. Given the difference in immune response to infection with wild-type and ΔrpoE1, we tested whether ΔrpoE1 confers protective immunity to wild-type challenge. Mice immunized with ΔrpoE1 completely resisted WT infection and had significantly higher serum titers of Brucella-specific IgG, IgG2a and IFN-γ after WT challenge relative to age-matched naïve mice. We conclude that immunization of BALB/c mice with the B. abortus GSR pathway mutant, ΔrpoE1, elicits an adaptive immune response that confers significant protective immunity against WT infection.

  11. 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

  12. 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

  13. Structure of the pyruvate dehydrogenase multienzyme complex E1 component from Escherichia coli at 1.85 A resolution.

    PubMed

    Arjunan, Palaniappa; Nemeria, Natalia; Brunskill, Andrew; Chandrasekhar, Krishnamoorthy; Sax, Martin; Yan, Yan; Jordan, Frank; Guest, John R; Furey, William

    2002-04-23

    The crystal structure of the recombinant thiamin diphosphate-dependent E1 component from the Escherichia coli pyruvate dehydrogenase multienzyme complex (PDHc) has been determined at a resolution of 1.85 A. The E. coli PDHc E1 component E1p is a homodimeric enzyme and crystallizes with an intact dimer in an asymmetric unit. Each E1p subunit consists of three domains: N-terminal, middle, and C-terminal, with all having alpha/beta folds. The functional dimer contains two catalytic centers located at the interface between subunits. The ThDP cofactors are bound in the "V" conformation in clefts between the two subunits (binding involves the N-terminal and middle domains), and there is a common ThDP binding fold. The cofactors are completely buried, as only the C2 atoms are accessible from solution through the active site clefts. Significant structural differences are observed between individual domains of E1p relative to heterotetrameric multienzyme complex E1 components operating on branched chain substrates. These differences may be responsible for reported alternative E1p binding modes to E2 components within the respective complexes. This paper represents the first structural example of a functional pyruvate dehydrogenase E1p component from any species. It also provides the first representative example for the entire family of homodimeric (alpha2) E1 multienzyme complex components, and should serve as a model for this class of enzymes. PMID:11955070

  14. 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.

  15. De-repression of RaRF-mediated RAR repression by adenovirus E1A in the nucleolus.

    PubMed

    Um, Soo-Jong; Youn, Hye Sook; Kim, Eun-Joo

    2014-02-21

    Transcriptional activity of the retinoic acid receptor (RAR) is regulated by diverse binding partners, including classical corepressors and coactivators, in response to its ligand retinoic acid (RA). Recently, we identified a novel corepressor of RAR called the retinoic acid resistance factor (RaRF) (manuscript submitted). Here, we report how adenovirus E1A stimulates RAR activity by associating with RaRF. Based on immunoprecipitation (IP) assays, E1A interacts with RaRF through the conserved region 2 (CR2), which is also responsible for pRb binding. The first coiled-coil domain of RaRF was sufficient for this interaction. An in vitro glutathione-S-transferase (GST) pull-down assay was used to confirm the direct interaction between E1A and RaRF. Further fluorescence microscopy indicated that E1A and RaRF were located in the nucleoplasm and nucleolus, respectively. However, RaRF overexpression promoted nucleolar translocation of E1A from the nucleoplasm. Both the RA-dependent interaction of RAR with RaRF and RAR translocation to the nucleolus were disrupted by E1A. RaRF-mediated RAR repression was impaired by wild-type E1A, but not by the RaRF binding-defective E1A mutant. Taken together, our data suggest that E1A is sequestered to the nucleolus by RaRF through a specific interaction, thereby leaving RAR in the nucleoplasm for transcriptional activation.

  16. 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.

  17. Compressive strength of carbon fibers

    SciTech Connect

    Prandy, J.M. ); Hahn, H.T. )

    1991-01-01

    Most composites are weaker in compression than in tension, which is due to the poor compressive strength of the load bearing fibers. The present paper discusses the compressive strengths and failure modes of 11 different carbon fibers: PAN-AS1, AS4, IM6, IM7, T700, T300, GY-30, pitch-75, ultra high modulus (UHM), high modulus (HM), and high strength (HS). The compressive strength was determined by embedding a fiber bundle in a transparent epoxy matrix and testing in compression. The resin allows for the containment and observation of failure during and after testing while also providing lateral support to the fibers. Scanning electron microscopy (SEM) was used to determine the global failure modes of the fibers.

  18. 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

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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

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

    PubMed Central

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

    2016-01-01

    Signaling by ubiquitination regulates virtually every cellular process in eukaryotes. Covalent attachment of ubiquitin to a substrate is catalyzed by the E1, E2 and E3 three-enzyme cascade 1, which links the C terminus of ubiquitin via an isopeptide bond mostly to the ε-amino group of a lysine of the substrate. 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 2. For example, many bacterial pathogens exploit ubiquitin signaling using virulence factors that function as E3 ligases, deubiquitinases 3 or as enzymes that directly attack ubiquitin 4. The bacterial pathogen Legionella pneumophila utilizes approximately 300 effectors that modulate diverse host processes to create a niche permissive for its replication in phagocytes 5. Here we demonstrate that members of the SidE effector family (SidEs) of L. pneumophila ubiquitinate multiple Rab small GTPases associated with the endoplasmic reticulum (ER). Moreover, we show that these proteins are capable of catalyzing ubiquitination without the need for the E1 and E2 enzymes. A putative mono ADP-ribosyltransferase (mART) motif critical for the ubiquitination activity is also essential for the role of SidEs in intracellular bacterial replication in a protozoan host. The E1/E2-independent ubiquitination catalyzed by these enzymes is energized by NAD which activates ubiquitin by the formation of ADP-ribosylated ubiquitin (ADPR-Ub). These results establish that ubiquitination can be catalyzed by a single enzyme whose activity does not require ATP. PMID:27049943

  7. 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.

  8. 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.

  9. The effect of hydrofluoric acid treatment of titanium surface on nanostructural and chemical changes and the growth of MC3T3-E1 cells.

    PubMed

    Lamolle, Sébastien F; Monjo, Marta; Rubert, Marina; Haugen, Håvard J; Lyngstadaas, Ståle P; Ellingsen, Jan E

    2009-02-01

    Fluoride-modification of dental titanium (Ti) implants is used to improve peri-implant bone growth and bone-to-implant contact and adhesion strength. In this study, the surface topography, chemistry and biocompatibility of polished Ti surfaces treated with hydrofluoric acid solution (HF) were studied. Murine osteoblasts (MC3T3-E1) were cultured on the different groups of Ti surfaces. Surfaces treated with HF had higher roughness, lower cytotoxicity level and better biocompatibility than controls. For short treatment times (40 and 90 s), fluorine was detected only within the first 5 nm of the surface layer (X-ray Photoemission Spectroscopy, XPS), whereas longer treatment time (120 and 150 s) caused fluoride ions to penetrate deeper (Secondary Ion Mass Spectrometry, SIMS). These results suggest that submerging Ti implants in a weak HF solution instigate time-dependant specific surface changes that are linked to the improved biocompatibility of these surfaces.

  10. Adenovirus type 5 early region 4 is responsible for E1A-induced p53-independent apoptosis.

    PubMed Central

    Marcellus, R C; Teodoro, J G; Wu, T; Brough, D E; Ketner, G; Shore, G C; Branton, P E

    1996-01-01

    In the absence of E1B, the 289- and 243-residue E1A products of human adenovirus type 5 induce p53-dependent apoptosis. However, our group has shown recently that the 289-residue E1A protein is also able to induce apoptosis by a p53-independent mechanism (J. G. Teodoro, G. C. Shore, and P. E. Branton, Oncogene 11:467-474, 1995). Preliminary results suggested that p53-independent cell death required expression of one or more additional adenovirus early gene products. Here we show that both the E1B 19-kDa protein and cellular Bcl-2 inhibit or significantly delay p53-independent apoptosis. Neither early region E2 or E3 appeared to be necessary for such cell death. Analysis of a series of E1A mutants indicated that mutations in the transactivation domain and other regions of E1A correlated with E1A-mediated transactivation of E4 gene expression. Furthermore, p53-deficient human SAOS-2 cells infected with a mutant which expresses E1B but none of the E4 gene products remained viable for considerably longer times than those infected with wild-type adenovirus type 5. In addition, an adenovirus vector lacking both E1 and E4 was unable to induce DNA degradation and cell killing in E1A-expressing cell lines. These data showed that an E4 product is essential for E1A-induced p53-independent apoptosis. PMID:8709247

  11. Role for intestinal CYP2E1 in alcohol-induced circadian gene-mediated intestinal hyperpermeability.

    PubMed

    Forsyth, Christopher B; Voigt, Robin M; Shaikh, Maliha; Tang, Yueming; Cederbaum, Arthur I; Turek, Fred W; Keshavarzian, Ali

    2013-07-15

    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

  12. 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

  13. Biology of E1-Deleted Adenovirus Vectors in Nonhuman Primate Muscle

    PubMed Central

    Zoltick, Philip W.; Chirmule, Narendra; Schnell, Michael A.; Gao, Guang-ping; Hughes, Joseph V.; Wilson, James M.

    2001-01-01

    Adenovirus vectors have been studied as vehicles for gene transfer to skeletal muscle, an attractive target for gene therapies for inherited and acquired diseases. In this setting, immune responses to viral proteins and/or transgene products cause inflammation and lead to loss of transgene expression. A few studies in murine models have suggested that the destructive cell-mediated immune response to virally encoded proteins of E1-deleted adenovirus may not contribute to the elimination of transgene-expressing cells. However, the impact of immune responses following intramuscular administration of adenovirus vectors on transgene stability has not been elucidated in larger animal models such as nonhuman primates. Here we demonstrate that intramuscular administration of E1-deleted adenovirus vector expressing rhesus monkey erythropoietin or growth hormone to rhesus monkeys results in generation of a Th1-dependent cytotoxic T-cell response to adenovirus proteins. Transgene expression dropped significantly over time but was still detectable in some animals after 6 months. Systemic levels of adenovirus-specific neutralizing antibodies were generated, which blocked vector readministration. These studies indicate that the cellular and humoral immune response generated to adenovirus proteins, in the context of transgenes encoding self-proteins, hinders long-term transgene expression and readministration with first-generation vectors. PMID:11333904

  14. The ColE1 unidirectional origin acts as a polar replication fork pausing site.

    PubMed

    Viguera, E; Hernández, P; Krimer, D B; Boistov, A S; Lurz, R; Alonso, J C; Schvartzman, J B

    1996-09-13

    Co-orientation of replication origins is the most common organization found in nature for multimeric plasmids. Streptococcus pyogenes broad-host-range plasmid pSM19035 and Escherichia coli pPI21 are among the exceptions. pPI21, which is a derivative of pSM19035 and pBR322, has two long inverted repeats, each one containing a potentially active ColE1 unidirectional origin. Analysis of pPI21 replication intermediates (RIs) by two-dimensional agarose gel electrophoresis and electron microscopy revealed the accumulation of a specific RI containing a single internal bubble. The data obtained demonstrated that initiation of DNA replication occurred at a single origin in pPI21. Progression of the replicating fork initiated at either of the two potential origins was transiently stalled at the other inversely oriented silent ColE1 origin of the plasmid. The accumulated RIs, containing an internal bubble, occurred as a series of stereoisomers with different numbers of knots in their replicated portion. These observations provide one of the first functional explanations for the disadvantage of head-to-head plasmid multimers with respect to head-to-tail ones.

  15. A cascading activity-based probe sequentially targets E1-E2-E3 ubiquitin enzymes.

    PubMed

    Mulder, Monique P C; Witting, Katharina; Berlin, Ilana; Pruneda, Jonathan N; Wu, Kuen-Phon; Chang, Jer-Gung; Merkx, Remco; Bialas, Johanna; Groettrup, Marcus; Vertegaal, Alfred C O; Schulman, Brenda A; Komander, David; Neefjes, Jacques; El Oualid, Farid; Ovaa, Huib

    2016-07-01

    Post-translational modifications of proteins with ubiquitin (Ub) and ubiquitin-like modifiers (Ubls), orchestrated by a cascade of specialized E1, E2 and E3 enzymes, control a wide range of cellular processes. To monitor catalysis along these complex reaction pathways, we developed a cascading activity-based probe, UbDha. Similarly to the native Ub, upon ATP-dependent activation by the E1, UbDha can travel downstream to the E2 (and subsequently E3) enzymes through sequential trans-thioesterifications. Unlike the native Ub, at each step along the cascade, UbDha has the option to react irreversibly with active site cysteine residues of target enzymes, thus enabling their detection. We show that our cascading probe 'hops' and 'traps' catalytically active Ub-modifying enzymes (but not their substrates) by a mechanism diversifiable to Ubls. Our founder methodology, amenable to structural studies, proteome-wide profiling and monitoring of enzymatic activity in living cells, presents novel and versatile tools to interrogate Ub and Ubl cascades. PMID:27182664

  16. [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.

  17. Hypothesis: hypersensitive plasmid copy number control for ColE1.

    PubMed Central

    Ehrenberg, M

    1996-01-01

    Initiation of replication of the plasmid ColE1 is primed by the cis-acting RNA II. Copy numbers are regulated by inhibition of RNA II by the antisense RNA I, whose concentration is proportional to the plasmid concentration. This inhibition is enhanced by a protein. Rom, and takes place during a time set by the transcription of 250 bases of the gene for RNA II. When this transcription is dominated by several steps of about equal duration, the probability for RNA II to prime DNA replication is approximately determined by e-constant[RNA I]. For large values of the "constant" small changes in [RNA I] give large variations in the priming probability. It is shown, first, that this type of mechanism can reduce the rate of plasmid loss and enable single copies of ColE1 to duplicate at a well-defined time in the cell cycle; second, that when the rate of initiation of transcription of RNA II increases, plasmid losses decrease and the distribution of single copy duplication times becomes narrower; third, that the action of Rom may further reduce plasmid losses and further narrow the distribution of duplication times in the single-copy case. PMID:8770193

  18. Buserelin acetate microparticle dispersion effects drug release and plasma E(1) levels.

    PubMed

    Usami, Makiko; Misawa, Kazumasa; Yagi, Naomi; Sekikawa, Hitoshi; Nabeshima, Toshitaka

    2007-07-18

    We investigated the effect of different dispersion methods on release behavior and efficacy onset following microparticle administration of buserelin acetate (BA) sustained-release injection. In this in vitro release study, the initial dispersion of BA increased with increased stirring speed (p<0.01). Stability of BA was studied over 7 days after BA release. The initial BA release rate was higher (p<0.01) after a 1-min vibration dispersion method (VDM) using a test tube mixer (2000 rpm) compared with the standard dispersion method (SDM) by hand. Without shaking, powder aggregation was observed, and BA release was lower than in either the SDM or VDM methods. In this study using 4-week-old Sprague-Dawley female rats, the initial plasma estrone (E(1)) concentrations were lower (p<0.05) in the VDM method than in the SDM method. Observations by optical microscope and scanning microscope showed no change in microparticle shape or distribution of size induced by SDM, VDM or the ultrasonication dispersion method. These results suggest that different dispersion methods do not change the shape and distribution of microparticle size, but clearly change the BA release rate and the transition in plasma E(1) concentrations that can affect drug efficacy. PMID:17398044

  19. 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

  20. Watercress has no Importance for the elimination of ethanol by CYP2E1 inhibition.

    PubMed

    Desager, Jean-Pierre; Golnez, Jean-Luc; De Buck, Charlotte; Horsmans, Yves

    2002-09-01

    Watercress, a cruciferous vegetable, is known to inhibit the metabolism of several CYP2E1 substrates such as paracetamol and chlorzoxazone. Since ethanol and its metabolite, acetaldehyde, are CYP2E1 substrates, the influence of watercress on ethanol and acetaldehyde was investigated in healthy human volunteers. According to a randomized cross-over design, ethanol and acetaldehyde pharmacokinetic parameters were determined in 9 persons at 3 occasions: without watercress and after watercress ingestion preceding ethanol consumption from 1 or 10.5 hr, respectively. Ethanol tmax occurred significantly later when watercress was ingested 1 hr before ethanol ingestion. Likewise, acetaldehyde Cmax was significantly higher whereas acetaldehyde AUCs were increased by watercress but not significantly. All other ethanol and acetaldehyde pharmacokinetic parameters were similar between the 3 treatments. In healthy volunteers, no major watercress effect was observed on ethanol clearance but a weak inhibiting effect on acetaldehyde metabolism is possible. Ethanol absorption is also delayed by single ingestion of watercress immediately preceding ethanol consumption.

  1. Poly(L-lactide) crystallization topography directs MC3T3-E1 cells response.

    PubMed

    Li, Wenqiang; Lu, Lu; Jiao, Yanpeng; Zhang, Chaowen; Zhou, Changren

    2016-09-01

    Biomaterial surface topography significantly influences cellular form and function. Using poly(L-lactic acid) films with normal spherulites, banded spherulites, and amorphous surfaces as model substrates, we conducted a systematic assessment of the role for polymer crystallization induced surface morphologies on cell growth and contact guidance. Microscopy and image analysis showed that the MC3T3-E1 cells spread out in a random fashion on the amorphous substrate. At 24 h post-seeding, MC3T3-E1 cells on both types of spherulite surfaces were elongated and aligned along the spherulite radius direction. For the banded spherulite surface with radial stripes and coupling annular grooves, the cell orientation and cell nuclear localization were related to the grooves structure. With increasing time, this orientation preference was weaker. These results demonstrate that the patterning of polymer crystallization structure provide important signals for guiding cells to exhibit characteristic orientation and morphology especially in the early stages of regeneration. PMID:27376548

  2. 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.

  3. The Human Adenovirus Type 5 E4orf6/E1B55K E3 Ubiquitin Ligase Complex Can Mimic E1A Effects on E2F

    PubMed Central

    Dallaire, Frédéric; Schreiner, Sabrina; Blair, G. Eric; Dobner, Thomas; Branton, Philip E.

    2015-01-01

    ABSTRACT The human adenovirus E4orf6/E1B55K E3 ubiquitin ligase is well known to promote viral replication by degrading an increasing number of cellular proteins that inhibit the efficient production of viral progeny. We report here a new function of the adenovirus 5 (Ad5) viral ligase complex that, although at lower levels, mimics effects of E1A products on E2F transcription factors. When expressed in the absence of E1A, the E4orf6 protein in complex with E1B55K binds E2F, disrupts E2F/retinoblastoma protein (Rb) complexes, and induces hyperphosphorylation of Rb, leading to induction of viral and cellular DNA synthesis as well as stimulation of early and late viral gene expression and production of viral progeny of E1/E3-defective adenovirus vectors. These new and previously undescribed functions of the E4orf6/E1B55K E3 ubiquitin ligase could play an important role in promoting the replication of wild-type viruses. IMPORTANCE During the course of work on the adenovirus E3 ubiquitin ligase formed by the viral E4orf6 and E1B55K proteins, we found, very surprisingly, that expression of these species was sufficient to permit low levels of replication of an adenovirus vector lacking E1A, the central regulator of infection. E1A products uncouple E2F transcription factors from Rb repression complexes, thus stimulating viral gene expression and cell and viral DNA synthesis. We found that the E4orf6/E1B55K ligase mimics these functions. This finding is of significance because it represents an entirely new function for the ligase in regulating adenovirus replication. PMID:27303679

  4. The Human Adenovirus Type 5 E4orf6/E1B55K E3 Ubiquitin Ligase Complex Can Mimic E1A Effects on E2F.

    PubMed

    Dallaire, Frédéric; Schreiner, Sabrina; Blair, G Eric; Dobner, Thomas; Branton, Philip E; Blanchette, Paola

    2016-01-01

    The human adenovirus E4orf6/E1B55K E3 ubiquitin ligase is well known to promote viral replication by degrading an increasing number of cellular proteins that inhibit the efficient production of viral progeny. We report here a new function of the adenovirus 5 (Ad5) viral ligase complex that, although at lower levels, mimics effects of E1A products on E2F transcription factors. When expressed in the absence of E1A, the E4orf6 protein in complex with E1B55K binds E2F, disrupts E2F/retinoblastoma protein (Rb) complexes, and induces hyperphosphorylation of Rb, leading to induction of viral and cellular DNA synthesis as well as stimulation of early and late viral gene expression and production of viral progeny of E1/E3-defective adenovirus vectors. These new and previously undescribed functions of the E4orf6/E1B55K E3 ubiquitin ligase could play an important role in promoting the replication of wild-type viruses. IMPORTANCE During the course of work on the adenovirus E3 ubiquitin ligase formed by the viral E4orf6 and E1B55K proteins, we found, very surprisingly, that expression of these species was sufficient to permit low levels of replication of an adenovirus vector lacking E1A, the central regulator of infection. E1A products uncouple E2F transcription factors from Rb repression complexes, thus stimulating viral gene expression and cell and viral DNA synthesis. We found that the E4orf6/E1B55K ligase mimics these functions. This finding is of significance because it represents an entirely new function for the ligase in regulating adenovirus replication. PMID:27303679

  5. 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...

  6. 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

  7. 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.

  8. 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

  9. 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

  10. 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....

  11. Hepatitis C Virus E1 and E2 Proteins Used as Separate Immunogens Induce Neutralizing Antibodies with Additive Properties.

    PubMed

    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.

  12. Immunoelectron microscopic localization of the ubiquitin-activating enzyme E1 in HepG2 cells.

    PubMed Central

    Schwartz, A L; Trausch, J S; Ciechanover, A; Slot, J W; Geuze, H

    1992-01-01

    As the first enzyme in the ubiquitin system the ubiquitin-activating enzyme E1 plays a pivotal role in all pathways of protein ubiquitination. In an effort to learn more about the cell biology of this pathway, we have purified the 110-kDa enzyme to homogeneity and generated a panel of distinct monoclonal antibodies to it. Using quantitative electron microscopic immunolocalization with these anti-E1 monoclonal antibodies, we find that E1 is abundant both within the cytoplasm and nucleus. Within the cytoplasm, E1 was found throughout the cytoplasmic volume as well as enriched along the cytoplasmic face of the rough endoplasmic reticulum and associated with the dense material along the desmosomal junctions. E1 was also found associated with the cytoplasmic surface of endosomal/lysosomal vacuoles. Interestingly, E1 was also found within the mitochondria. The lumen of rough endoplasmic reticulum, Golgi complex, endosomes, and lysosomes were negative. The specific localization of E1 to distinct subcellular organelles suggests that E1 may play multiple physiological roles within the cell. Images PMID:1376922

  13. 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.

  14. The Cellular Protein Complex Associated with a Transforming Region of E1A Contains c-MYC

    PubMed Central

    Vijayalingam, S.; Subramanian, T.; Zhao, Ling-jun

    2015-01-01

    ABSTRACT The cell-transforming activity of human adenovirus 5 (hAd5) E1A is mediated by the N-terminal half of E1A, which interacts with three different major cellular protein complexes, p300/CBP, TRRAP/p400, and pRb family members. Among these protein interactions, the interaction of pRb family proteins with conserved region 2 (CR2) of E1A is known to promote cell proliferation by deregulating the activities of E2F family transcription factors. The functional consequences of interaction with the other two protein complexes in regulating the transforming activity of E1A are not well defined. Here, we report that the E1A N-terminal region also interacted with the cellular proto-oncoprotein c-MYC and the homolog of enhancer of yellow 2 (ENY2). Our results suggested that these proteins interacted with an essential E1A transforming domain spanning amino acid residues 26 to 35 which also interacted with TRRAP and p400. Small interfering RNA (siRNA)-mediated depletion of TRRAP reduced c-MYC interaction with E1A, while p400 depletion did not. In contrast, depletion of TRRAP enhanced ENY2 interaction with E1A, suggesting that ENY2 and TRRAP may interact with E1A in a competitive manner. The same E1A region additionally interacted with the constituents of a deubiquitinase complex consisting of USP22, ATXN7, and ATXN7L3 via TRRAP. Acute short hairpin RNA (shRNA)-mediated depletion of c-MYC reduced the E1A transforming activity, while depletion of ENY2 and MAX did not. These results suggested that the association of c-MYC with E1A may, at least partially, play a role in the E1A transformation activity, independently of MAX. IMPORTANCE The transforming region of adenovirus E1A consists of three short modules which complex with different cellular protein complexes. The mechanism by which one of the transforming modules, CR2, promotes cell proliferation, through inactivating the activities of the pRb family proteins, is better understood than the activities of the other domains

  15. 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.

  16. Lifting strength in two-person teamwork.

    PubMed

    Lee, Tzu-Hsien

    2016-01-01

    This study examined the effects of lifting range, hand-to-toe distance, and lifting direction on single-person lifting strengths and two-person teamwork lifting strengths. Six healthy males and seven healthy females participated in this study. Two-person teamwork lifting strengths were examined in both strength-matched and strength-unmatched groups. Our results showed that lifting strength significantly decreased with increasing lifting range or hand-to-toe distance. However, lifting strengths were not affected by lifting direction. Teamwork lifting strength did not conform to the law of additivity for both strength-matched and strength-unmatched groups. In general, teamwork lifting strength was dictated by the weaker of the two members, implying that weaker members might be exposed to a higher potential danger in teamwork exertions. To avoid such overexertion in teamwork, members with significantly different strength ability should not be assigned to the same team.

  17. Ultrasound stimulation increases proliferation of MC3T3-E1 preosteoblast-like cells

    PubMed Central

    2014-01-01

    Background Mechanical stimulation of bone increases bone mass and fracture healing, at least in part, through increases in proliferation of osteoblasts and osteoprogenitor cells. Researchers have previously performed in vitro studies of ultrasound-induced osteoblast proliferation but mostly used fixed ultrasound settings and have reported widely varying and inconclusive results. Here we critically investigated the effects of the excitation parameters of low-intensity pulsed ultrasound (LIPUS) stimulation on proliferation of MC3T3-E1 preosteoblastic cells in monolayer cultures. Methods We used a custom-designed ultrasound exposure system to vary the key ultrasound parameters—intensity, frequency and excitation duration. MC3T3-E1 cells were seeded in 12-well cell culture plates. Unless otherwise specified, treated cells, in groups of three, were excited twice for 10 min with an interval of 24 h in between after cell seeding. Proliferation rates of these cells were determined using BrdU and MTS assays 24 h after the last LIPUS excitation. All data are presented as the mean ± standard error. The statistical significance was determined using Student's two-sample two-tailed t tests. Results Using discrete LIPUS intensities ranging from 1 to 500 mW/cm2 (SATA, spatial average-temporal average), we found that approximately 75 mW/cm2 produced the greatest increase in osteoblast proliferation. Ultrasound exposures at higher intensity (approximately 465 mW/cm2) significantly reduced proliferation in MC3T3-E1 cells, suggesting that high-intensity pulsed ultrasound may increase apoptosis or loss of adhesion in these cells. Variation in LIPUS frequency from 0.5 MHz to 5 MHz indicated that osteoblast proliferation rate was not frequency dependent. We found no difference in the increase in proliferation rate if LIPUS was applied for 30 min/day or 10 min/day, indicating a habituation response. Conclusion This study concludes that a short-term stimulation with optimum intensity

  18. 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

  19. 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

  20. A novel CRM1-dependent nuclear export signal in adenoviral E1A protein regulated by phosphorylation.

    PubMed

    Jiang, Hong; Olson, Melissa V; Medrano, Diana R; Lee, Ok-Hee; Xu, Jing; Piao, Yuji; Alonso, Marta M; Gomez-Manzano, Candelaria; Hung, Mien-Chie; Yung, W K Alfred; Fueyo, Juan

    2006-12-01

    Adenoviral E1A is a versatile protein that can reprogram host cells for efficient viral replication. The nuclear import of E1A is mediated by a nuclear localization signal; however, whether E1A can be actively exported from the nucleus is unknown. We first reported a CRM1-dependent nuclear export signal (NES) in E1A that is conserved in the group C adenoviruses. We showed that CRM1 and E1A coimmunoprecipitated and that blockage of CRM1 function by leptomycin B or small interfering RNA resulted in the nuclear localization of E1A. Through mutational analyses, we identified an active canonical NES element within the E1A protein spanning amino acids 70-80. We further demonstrated that phosphorylation of adjacent serine (S)89 resulted in the cytoplasmic accumulation of E1A. Interestingly, coincident with the accumulation of cells in the S/G2/M phase and histone H1 phosphorylation, E1A was relocated to the cytoplasm at the late stage of the viral cycle, which was blocked by the CDC2/CDK2 inhibitor roscovitine. Importantly, titration of the progenies of the viruses in infected cells showed that the replication efficiency of the NES mutant adenovirus was up to 500-fold lower than that of the wild-type adenovirus. Collectively, our data demonstrate the existence of a NES in E1A that is modulated by the phosphorylation of the S89 residue and the NES plays a role for an efficient viral replication in the host cells.

  1. MicroRNA-16 modulates HuR regulation of cyclin E1 in breast cancer cells.

    PubMed

    Guo, Xun; Connick, Melanie C; Vanderhoof, Jennifer; Ishak, Mohammad-Ali; Hartley, Rebecca S

    2015-03-30

    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.

  2. Inhibition of Human Papillomavirus DNA Replication by an E1-Derived p80/UAF1-Binding Peptide

    PubMed Central

    Lehoux, Michaël; Fradet-Turcotte, Amélie; Lussier-Price, Mathieu; Omichinski, James G.

    2012-01-01

    The papillomavirus E1 helicase is recruited by E2 to the viral origin, where it assembles into a double hexamer that orchestrates replication of the viral genome. We previously identified the cellular WD40 repeat-containing protein p80/UAF1 as a novel interaction partner of E1 from anogenital human papillomavirus (HPV) types. p80 was found to interact with the first 40 residues of HPV type 31 (HPV31) E1, and amino acid substitutions within this domain abrogated the maintenance of the viral episome in keratinocytes. In this study, we report that these p80-binding substitutions reduce by 70% the ability of E1 to support transient viral DNA replication without affecting its interaction with E2 and assembly at the origin in vivo. Microscopy studies revealed that p80 is relocalized from the cytoplasm to discrete subnuclear foci by E1 and E2. Chromatin immunoprecipitation assays further revealed that p80 is recruited to the viral origin in an E1- and E2-dependent manner. Interestingly, overexpression of a 40-amino-acid-long p80-binding peptide, derived from HPV31 E1, was found to inhibit viral DNA replication by preventing the recruitment of endogenous p80 to the origin. Mutant peptides defective for p80 interaction were not inhibitory, demonstrating the specificity of this effect. Characterization of this E1 peptide by nuclear magnetic resonance (NMR) showed that it is intrinsically disordered in solution, while mapping studies indicated that the WD repeats of p80 are required for E1 interaction. These results provide additional evidence for the requirement for p80 in anogenital HPV DNA replication and highlight the potential of E1-p80 interaction as a novel antiviral target. PMID:22278251

  3. Inhibition of human papillomavirus DNA replication by an E1-derived p80/UAF1-binding peptide.

    PubMed

    Lehoux, Michaël; Fradet-Turcotte, Amélie; Lussier-Price, Mathieu; Omichinski, James G; Archambault, Jacques

    2012-04-01

    The papillomavirus E1 helicase is recruited by E2 to the viral origin, where it assembles into a double hexamer that orchestrates replication of the viral genome. We previously identified the cellular WD40 repeat-containing protein p80/UAF1 as a novel interaction partner of E1 from anogenital human papillomavirus (HPV) types. p80 was found to interact with the first 40 residues of HPV type 31 (HPV31) E1, and amino acid substitutions within this domain abrogated the maintenance of the viral episome in keratinocytes. In this study, we report that these p80-binding substitutions reduce by 70% the ability of E1 to support transient viral DNA replication without affecting its interaction with E2 and assembly at the origin in vivo. Microscopy studies revealed that p80 is relocalized from the cytoplasm to discrete subnuclear foci by E1 and E2. Chromatin immunoprecipitation assays further revealed that p80 is recruited to the viral origin in an E1- and E2-dependent manner. Interestingly, overexpression of a 40-amino-acid-long p80-binding peptide, derived from HPV31 E1, was found to inhibit viral DNA replication by preventing the recruitment of endogenous p80 to the origin. Mutant peptides defective for p80 interaction were not inhibitory, demonstrating the specificity of this effect. Characterization of this E1 peptide by nuclear magnetic resonance (NMR) showed that it is intrinsically disordered in solution, while mapping studies indicated that the WD repeats of p80 are required for E1 interaction. These results provide additional evidence for the requirement for p80 in anogenital HPV DNA replication and highlight the potential of E1-p80 interaction as a novel antiviral target. PMID:22278251

  4. Inhibition of human papillomavirus DNA replication by an E1-derived p80/UAF1-binding peptide.

    PubMed

    Lehoux, Michaël; Fradet-Turcotte, Amélie; Lussier-Price, Mathieu; Omichinski, James G; Archambault, Jacques

    2012-04-01

    The papillomavirus E1 helicase is recruited by E2 to the viral origin, where it assembles into a double hexamer that orchestrates replication of the viral genome. We previously identified the cellular WD40 repeat-containing protein p80/UAF1 as a novel interaction partner of E1 from anogenital human papillomavirus (HPV) types. p80 was found to interact with the first 40 residues of HPV type 31 (HPV31) E1, and amino acid substitutions within this domain abrogated the maintenance of the viral episome in keratinocytes. In this study, we report that these p80-binding substitutions reduce by 70% the ability of E1 to support transient viral DNA replication without affecting its interaction with E2 and assembly at the origin in vivo. Microscopy studies revealed that p80 is relocalized from the cytoplasm to discrete subnuclear foci by E1 and E2. Chromatin immunoprecipitation assays further revealed that p80 is recruited to the viral origin in an E1- and E2-dependent manner. Interestingly, overexpression of a 40-amino-acid-long p80-binding peptide, derived from HPV31 E1, was found to inhibit viral DNA replication by preventing the recruitment of endogenous p80 to the origin. Mutant peptides defective for p80 interaction were not inhibitory, demonstrating the specificity of this effect. Characterization of this E1 peptide by nuclear magnetic resonance (NMR) showed that it is intrinsically disordered in solution, while mapping studies indicated that the WD repeats of p80 are required for E1 interaction. These results provide additional evidence for the requirement for p80 in anogenital HPV DNA replication and highlight the potential of E1-p80 interaction as a novel antiviral target.

  5. 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

  6. 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

  7. The use of laser altimetry data in Chang'E-1 precision orbit determination

    NASA Astrophysics Data System (ADS)

    Chang, Sheng-Qi; Huang, Yong; Li, Pei-Jia; Hu, Xiao-Gong; Fan, Min

    2016-09-01

    Accurate altimetric measurement not only can be applied to the calculation of a topography model but also can be used to improve the quality of the orbit reconstruction in the form of crossovers. Altimetry data from the Chang'E-1 (CE-1) laser altimeter are analyzed in this paper. The differences between the crossover constraint equation in the form of height discrepancies and in the form of minimum distances are mainly discussed. The results demonstrate that the crossover constraint equation in the form of minimum distances improves the CE-1 orbit precision. The overlap orbit performance has increased ∼ 30% compared to the orbit using only tracking data. External assessment using the topography model also shows orbit improvement. The results will be helpful for recomputing ephemeris and improving the CE-1 topography model.

  8. 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

  9. Application of ozonation process in industrial wastewaters: textile, kraft E1 and whey effluents.

    PubMed

    Assalin, M R; Almeida, E S; Rosa, M A; Moraes, S G; Duran, N

    2004-08-01

    A large variety of organic and inorganic compounds can be found in wastewater from industrial processes. In this work, Advanced Oxidative Processes (AOPs) have been applied for the control of water pollution and the ozonation of different effluents was investigated. Wastewater from textile, kraft E1 and cheese manufacturing processes were chosen as examples of industrial effluents. The efficiency of substrate mineralization has been comparatively analyzed by the decrease in total organic carbon (TOC), color, and toxicity. The results revealed that the ozonation process can be a method for decolorization of effluent, but it is not effective for TOC reduction. The whey effluent was the most recalcitrant wastewater for ozone treatment which produced no TOC removal.

  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. 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.

  12. A human papilloma virus type 11 transcript encoding an E1--E4 protein.

    PubMed

    Nasseri, M; Hirochika, R; Broker, T R; Chow, L T

    1987-08-01

    The human papilloma virus (HPV) associated with a genital wart (condyloma acuminatum) was determined to be type 11. The majority of the viral DNA molecules were monomeric circles present in the cells at high copy number, as demonstrated by one- and two-dimensional agarose gell electrophoretic separation followed by Southern blot analysis. A cDNA library in phage lambda gt11 was constructed from poly(A)-selected mRNA recovered from the tissue. Recombinant clones corresponding to the most abundant 1.2-kb viral mRNA species detected by Northern blot hybridization and by electron microscopic analysis of R loops were isolated and their nucleotide sequence was determined. Comparison to the prototype HPV-11 DNA sequence revealed that this message consisted of two exons. The promotor-proximal exon spanned nucleotides 716 through 847 and the distal exon included nucleotides 3325 through 4390 or 4392. The mRNAs were alternatively polyadenylated after either of these latter two sites, in both cases following a G and preceding a U residue. Fourteen or sixteen bases upstream from the poly(A) was the hexanucleotide AGUAAA, which apparently serves as the signal for cleavage and polyadenylation of the nascent message. The splice donor and acceptor sites conformed to the usual /GU. . .AG/pattern. The exons joined open reading frame (ORF) E1, which contributed the initiation codon and four additional triplets, to ORF E4, which specified 85 amino acids to encode a protein of 10,022 Da. The cDNA also contained the ORFs E5a and E5b toward the 3' end. The complete sequence of the cDNA revealed three single-base changes from the prototype HPV-11, two resulting in altered amino acids in E4. Neither affects the coding potential of the overlapping E2 ORF. The function of the E1--E4 protein is unknown. PMID:2887066

  13. 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

  14. 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

  15. Communication between Thiamin Cofactors in the Escherichia coli Pyruvate Dehydrogenase Complex E1 Component Active Centers

    PubMed Central

    Nemeria, Natalia S.; Arjunan, Palaniappa; Chandrasekhar, Krishnamoorthy; Mossad, Madouna; Tittmann, Kai; Furey, William; Jordan, Frank

    2010-01-01

    Kinetic, spectroscopic, and structural analysis tested the hypothesis that a chain of residues connecting the 4′-aminopyrimidine N1′ atoms of thiamin diphosphates (ThDPs) in the two active centers of the Escherichia coli pyruvate dehydrogenase complex E1 component provides a signal transduction pathway. Substitution of the three acidic residues (Glu571, Glu235, and Glu237) and Arg606 resulted in impaired binding of the second ThDP, once the first active center was filled, suggesting a pathway for communication between the two ThDPs. 1) Steady-state kinetic and fluorescence quenching studies revealed that upon E571A, E235A, E237A, and R606A substitutions, ThDP binding in the second active center was affected. 2) Analysis of the kinetics of thiazolium C2 hydrogen/deuterium exchange of enzyme-bound ThDP suggests half-of-the-sites reactivity for the E1 component, with fast (activated site) and slow exchanging sites (dormant site). The E235A and E571A variants gave no evidence for the slow exchanging site, indicating that only one of two active sites is filled with ThDP. 3) Titration of the E235A and E237A variants with methyl acetylphosphonate monitored by circular dichroism suggested that only half of the active sites were filled with a covalent predecarboxylation intermediate analog. 4) Crystal structures of E235A and E571A in complex with ThDP revealed the structural basis for the spectroscopic and kinetic observations and showed that either substitution affects cofactor binding, despite the fact that Glu235 makes no direct contact with the cofactor. The role of the conserved Glu571 residue in both catalysis and cofactor orientation is revealed by the combined results for the first time. PMID:20106967

  16. Characterization of the colistin (polymyxin E1 and E2) biosynthetic gene cluster.

    PubMed

    Tambadou, Fatoumata; Caradec, Thibault; Gagez, Anne-Laure; Bonnet, Antoine; Sopéna, Valérie; Bridiau, Nicolas; Thiéry, Valérie; Didelot, Sandrine; Barthélémy, Cyrille; Chevrot, Romain

    2015-05-01

    Colistin is a mixture of polymyxin E1 and E2, bactericidal pentacationic lipopeptides used to treat infections caused by Gram-negative pathogens such as Pseudomonas aeruginosa and Klebsiella pneumoniae. Industrial production of colistin is obtained by a fermentation process of the natural producer Paenibacillus polymyxa var colistinus. NonRibosomal peptide synthetases (NRPS) coding the biosynthesis of polymyxins A, B and P have been recently described, rendering thereof the improvement of their production possible. However, the colistin biosynthesis pathway was not published so far. In this study, a Paenibacillus alvei has been identified by biochemical (Api 50 CH system) and molecular (16S rDNA sequencing) methods. Its culture supernatant displayed inhibitory activity against Gram-negative bacteria (P. aeruginosa, K. pneumoniae, Salmonella spp.). Two polymyxins, E1 and E2, were recovered from the supernatant and were characterized by high resolution LC-MS. A genomic library (960 clones) was constructed to identify the gene cluster responsible for biosynthesis of polymyxins. Selection of the clones harbouring the sequences of interest was obtained by a simple PCR-based screening. We used primers targeting NRPS sequences leading to the incorporation of amino acids present in polymyxins E. The sequences from three clones of interest were assembled on 50.4 kb. Thus, five open reading frames corresponding to a new NRPS gene cluster of 41 kb were identified. In silico, analyses revealed the presence of three NRPS implicated in the biosynthesis of polymyxins E. This work provides insightful information on colistin biosynthesis and might contribute to future drug developments in this group of antibiotics.

  17. 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.

  18. The torsional strength of wings

    NASA Technical Reports Server (NTRS)

    Burgess, C P

    1930-01-01

    This report describes a simple method for calculating the position of the elastic axis of a wing structure having any number of spars. It is shown that strong drag bracing near the top and bottom of a wing greatly increases the torsional strength. An analytical procedure for finding the contribution of the drag bracing to the torsional strength and stiffness is described, based upon the principle of least work, and involving only one unknown quantity. A coefficient for comparing the torsional rigidity of different wings is derived in this report.

  19. 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

  20. 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

  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

  2. 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.

  3. 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

  4. An interlaminar tension strength specimen

    NASA Technical Reports Server (NTRS)

    Jackson, Wade C.; Martin, Roderick H.

    1992-01-01

    This paper describes a technique to determine interlaminar tension strength, sigma(sub 3c) of a fiber reinforced composite material using a curved beam. The specimen was a unidirectional curved beam, bent 90 degrees, with straight arms. Attached to each arm was a hinged loading mechanism which was held by the grips of a tensile 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 loading arm length had no 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. Width was found to affect the value of sigma(sub 3c) only slightly. The wider specimens generally had a slightly lower strength since more material was under high stress, and hence, had a larger probability of containing a significant flaw.

  5. 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…

  6. 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…

  7. Adenovirus E1A Targets the DREF Nuclear Factor To Regulate Virus Gene Expression, DNA Replication, and Growth

    PubMed Central

    Radko, Sandi; Koleva, Maria; James, Kris M. D.; Jung, Richard; Mymryk, Joe S.

    2014-01-01

    ABSTRACT The adenovirus E1A gene is the first gene expressed upon viral infection. E1A remodels the cellular environment to maximize permissivity for viral replication. E1A is also the major transactivator of viral early gene expression and a coregulator of a large number of cellular genes. E1A carries out its functions predominantly by binding to cellular regulatory proteins and altering their activities. The unstructured nature of E1A enables it to bind to a large variety of cellular proteins and form new molecular complexes with novel functions. The C terminus of E1A is the least-characterized region of the protein, with few known binding partners. Here we report the identification of cellular factor DREF (ZBED1) as a novel and direct binding partner of E1A. Our studies identify a dual role for DREF in the viral life cycle. DREF contributes to activation of gene expression from all viral promoters early in infection. Unexpectedly, it also functions as a growth restriction factor for adenovirus as knockdown of DREF enhances virus growth and increases viral genome copy number late in the infection. We also identify DREF as a component of viral replication centers. E1A affects the subcellular distribution of DREF within PML bodies and enhances DREF SUMOylation. Our findings identify DREF as a novel E1A C terminus binding partner and provide evidence supporting a role for DREF in viral replication. IMPORTANCE This work identifies the putative transcription factor DREF as a new target of the E1A oncoproteins of human adenovirus. DREF was found to primarily localize with PML nuclear bodies in uninfected cells and to relocalize into virus replication centers during infection. DREF was also found to be SUMOylated, and this was enhanced in the presence of E1A. Knockdown of DREF reduced the levels of viral transcripts detected at 20 h, but not at 40 h, postinfection, increased overall virus yield, and enhanced viral DNA replication. DREF was also found to localize to

  8. 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.

  9. Induction of CYP2E1 in liver, kidney, brain and intestine during chronic ethanol administration and withdrawal: evidence that CYP2E1 possesses a rapid phase half-life of 6 hours or less.

    PubMed

    Roberts, B J; Shoaf, S E; Jeong, K S; Song, B J

    1994-12-15

    Controversy exists as to whether the induction of CYP2E1 by ethanol occurs via increased protein synthesis or protein stabilization. To address these issues in vivo, we chronically administered ethanol to rats and determined levels of immunoreactive CYP2E1 in liver, kidney, brain and upper gastro-intestinal tract (GI). Our data shows that chronic ethanol administration induces hepatic (5-6-fold over pair-fed controls) and extra-hepatic CYP2E1, an effect which is strikingly absent 12 hours after ethanol withdrawal. No changes in CYP2E1 mRNA were observed at any time, suggesting these changes are mainly post-translational at a blood ethanol concentration of 0.15% w/v. Our experimental data indicates that CYP2E1 possesses a half-life of 6 hours or less in the liver and is rapidly degraded following the removal of ethanol. This pattern of CYP2E1 turnover was also observed in other tissues, suggestive of a similar mode of regulation.

  10. 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

  11. 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…

  12. Design, synthesis and molecular docking of amide and urea derivatives as Escherichia coli PDHc-E1 inhibitors.

    PubMed

    He, Jun-Bo; Ren, Yan-Liang; Sun, Qiu-Shuang; You, Ge-Yun; Zhang, Li; Zou, Peng; Feng, Ling-Ling; Wan, Jian; He, Hong-Wu

    2014-06-15

    By targeting the ThDP binding site of Escherichia coli PDHc-E1, two new 'open-chain' classes of E. coli PDHc-E1 inhibitors, amide and urea derivatives, were designed, synthesized, and evaluated. The amide derivatives of compound 6d, with 4-NO2 in the benzene ring, showed the most potent inhibition of E. coli PDHc-E1. The urea derivatives displayed more potent inhibitory activity than the corresponding amide derivatives with the same substituent. Molecular docking studies confirmed that the urea derivatives have more potency due to the two hydrogen bonds formed by two NH of urea with Glu522. The docking results also indicate it might help us to design more efficient PDHc-E1 inhibitors that could interact with Glu522.

  13. The Role of CYP2E1 in Alcohol Metabolism and Sensitivity in the Central Nervous System

    PubMed Central

    Heit, Claire; Dong, Hongbin; Chen, Ying; Thompson, David C.; Deitrich, Richard A.; Vasiliou, Vasilis

    2015-01-01

    Ethanol consumption has effects on the central nervous system (CNS), manifesting as motor incoordination, sleep induction (hypnosis), anxiety, amnesia, and the reinforcement or aversion of alcohol consumption. Acetaldehyde (the direct metabolite of ethanol oxidation) contributes to many aspects of the behavioral effects of ethanol. Given acetaldehyde cannot pass through the blood brain barrier, its concentration in the CNS is primarily determined by local production from ethanol. Catalase and cytochrome P450 2E1(CYP2E1) represent the major enzymes in the CNS that catalyze ethanol oxidation. CYP2E1 is expressed abundantly within the microsomes of certain brain cells and is localized to particular brain regions. This chapter focuses on the discussion of CYP2E1 in ethanol metabolism in the CNS, covering topics including how it is regulated, where it is expressed and how it influences sensitivity to ethanol in the brain. PMID:23400924

  14. Regulation of hepatic sulfotransferase (SULT) 1E1 expression and effects on estrogenic activity in cystic fibrosis (CF)☆

    PubMed Central

    Falany, Charles N.; He, Dongning; Li, Li; Falany, Josie L.; Wilborn, Teresa W.; Kocarek, Thomas A.; Runge-Morris, Melissa

    2013-01-01

    Cystic fibrosis (CF) is a major genetic disease in Caucasians affecting 1 in 2500 newborns. Hepatobiliary pathology is a major cause of morbidity and mortality in CF second only to pulmonary disease. SULT1E1 activity is significantly elevated, generally 20–30-fold, in hepatocytes of mouse models of CF. SULT1E1 is responsible for the inactivation of β-estradiol (E2) at physiological concentrations via conjugation with sulfonate. The increase in SULT1E1 activity results in the alteration of E2-regulated protein expression in CF mouse liver. To investigate the mechanism by which the absence of CFTR in human cholangiocytes induces SULT1E1 expression in hepatocytes, a membrane-separated human MMNK-1 cholangiocyte and human HepG2 hepatocyte co-culture system was developed. The cystic fibrosis transmembrane conductance regulator (CFTR) is expressed in bile duct cholangiocytes but not hepatocytes, whereas SULT1E1 is expressed in hepatocytes but not cholangiocytes. CFTR expression in MMNK-1 cells was inhibited with siRNA by >90% as determined by immunoblot and immunohistochemical analysis. Control and CFTR-siRNA-MMNK-1 cells were co-cultured with HepG2 cells in a Transwell membrane-separated system. After 8 h of co-culture, HepG2 cells were removed from exposure to MMNK-1 cells and placed in fresh medium. After 24–48 h, expression of SULT1E1 and selected E2-regulated proteins was analyzed in the HepG2 cells. Results demonstrated that SULT1E1 message and activity were selectively induced in HepG2 cells co-cultured with CFTR-deficient MMNK-1 cells. The expression of E2-regulated proteins (IGF-1, GST-P1 and carbonic anhydrase II) was also altered in response to decreased E2 levels. Thus, the loss of CFTR activity in cholangiocytes stimulates the expression of SULT1E1 in hepatocytes by a paracrine mechanism. SULT1E1 expression in HepG2 cells is inducible by sterol mediated liver-X-receptor (LXR) activation although not by progestins that induce SULT1E1 in the endometrium

  15. 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

  16. Synergistic effects of AKAP95, Cyclin D1, Cyclin E1, and Cx43 in the development of rectal cancer

    PubMed Central

    Qi, Fengjie; Yuan, Yangyang; Zhi, Xuehong; Huang, Qian; Chen, Yuexin; Zhuang, Wenxin; Zhang, Dengcheng; Teng, Bogang; Kong, Xiangyu; Zhang, Yongxing

    2015-01-01

    Objective: To explore the expression of A-kinase anchor protein 95 (AKAP95), Cyclin D1, Cyclin E1, and Connexin43 (Cx43) in rectal cancer tissues and assess the associations between each of the proteins and pathological parameters, as well as their inter-relationships. Methods: AKAP95, Cyclin D1, Cyclin E1, and Cx43 protein expression rates were evaluated by immunohistochemistry in 50 rectal cancer specimens and 16 pericarcinoma tissues. Results: The positive rates of AKAP95, Cyclin E1, and Cyclin D1 proteins were 54.00 vs. 18.75%, 62.00 vs. 6.25%, and 72.00 vs. 31.25% in rectal cancer specimens and pericarcinoma tissues, respectively, representing statistically significant differences (P < 0.05). The positive rate of Cx43 protein expression in rectal cancer tissues was 44.00% and 62.50% in pericarcinoma tissues, and the difference between them was not significant (P > 0.05). No significant associations were found between protein expression of AKAP95, Cyclin E1, Cyclin D1, and Cx43, and the degree of differentiation, histological type, and lymph node metastasis of rectal cancer (P > 0.05). However, significant correlations were obtained between the expression rates of AKAP95 and Cyclin E1, Cyclin E1 and Cyclin D1, Cyclin E1 and Cx43 protein, and Cyclin D1 and Cx43, respectively (P < 0.05). Conclusion: AKAP95, Cyclin E1, and Cyclin D1 protein expression rates were significantly higher in rectal cancer tissues compared with pericarcinoma samples, suggesting an association between these proteins and the development and progression of rectal cancer. In addition, the significant correlations between the proteins (AKAP95 and Cyclin E1, Cyclin E1 and Cyclin D1, Cyclin E1 and Cx43 protein, and Cyclin D1 and Cx43) indicate the possible synergistic effects of these factors in the development and progression of rectal cancer. PMID:25973052

  17. 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 (2.92 Mb) 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

  18. pRB-E2F1 complexes are resistant to adenovirus E1A-mediated disruption.

    PubMed

    Seifried, L A; Talluri, S; Cecchini, M; Julian, L M; Mymryk, J S; Dick, F A

    2008-05-01

    Disruption of pRB-E2F interactions by E1A is a key event in the adenoviral life cycle that drives expression of early viral transcription and induces cell cycle progression. This function of E1A is complicated by E2F1, an E2F family member that controls multiple processes besides proliferation, including apoptosis and DNA repair. Recently, a second interaction site in pRB that only contacts E2F1 has been discovered, allowing pRB to control proliferation separately from other E2F1-dependent activities. Based on this new insight into pRB-E2F1 regulation, we investigated how E1A affects control of E2F1 by pRB. Our data reveal that pRB-E2F1 interactions are resistant to E1A-mediated disruption. Using mutant forms of pRB that selectively force E2F1 to bind through only one of the two binding sites on pRB, we determined that E1A is unable to disrupt E2F1's unique interaction with pRB. Furthermore, analysis of pRB-E2F complexes during adenoviral infection reveals the selective maintenance of pRB-E2F1 interactions despite the presence of E1A. Our experiments also demonstrate that E2F1 functions to maintain cell viability in response to E1A expression. This suggests that adenovirus E1A's seemingly complex mechanism of disrupting pRB-E2F interactions provides selectivity in promoting viral transcription and cell cycle advancement, while maintaining cell viability.

  19. Structure of Pyrazole Derivatives Impact their Affinity, Stoichiometry, and Cooperative Interactions for CYP2E1 Complexes

    PubMed Central

    Hartman, Jessica H.; Bradley, Amber M.; Laddusaw, Ryan M; Perry, Martin D.; Miller, Grover P.

    2013-01-01

    CYP2E1 plays a critical role in detoxication and carcinogenic activation of drugs, pollutants, and dietary compounds; however, these metabolic processes can involve poorly characterized cooperative interactions that compromise the ability to understand and predict CYP2E1 metabolism. Herein, we employed an array of ten azoles with an emphasis on pyrazoles to establish the selectivity of catalytic and cooperative CYP2E1 sites through binding and catalytic studies. Spectral binding studies for monocyclic azoles suggested two binding events, while bicyclic azoles suggested one. Pyrazole had moderate affinity toward the CYP2E1 catalytic site that improved when a methyl group was introduced at either position 3 or 4. The presence of methyl groups simultaneously at positions 3 and 5 blocked binding, and a phenyl group at position 3 did not improve binding affinity. In contrast, pyrazole fusion to a benzene or cyclohexane ring greatly increased affinity. The consequences of these binding events on CYP2E1 catalysis were studied through inhibition studies with 4-nitrophenol, a substrate known to bind both sites. Most pyrazoles shared a common mixed cooperative inhibition mechanism in which pyrazole binding rescued CYP2E1 from substrate inhibition. Overall, inhibitor affinities toward the CYP2E1 catalytic site were similar to those reported in binding studies, and the same trend was observed for binding at the cooperative site. Taken together, these studies identified key structural determinants in the affinity and stoichiometry of azole interactions with CYP2E1 and consequences on catalysis that further advance an understanding of the relationship between structure and function for this enzyme. PMID:23811196

  20. The adenovirus E1A protein targets the SAGA but not the ADA transcriptional regulatory complex through multiple independent domains.

    PubMed

    Shuen, Michael; Avvakumov, Nikita; Walfish, Paul G; Brandl, Chris J; Mymryk, Joe S

    2002-08-23

    Expression of the adenovirus E1A protein in the simple eukaryote Saccharomyces cerevisiae inhibits growth. We tested four regions of E1A that alter growth and transcription in mammalian cells for their effects in yeast when expressed as fusions to the Gal4p DNA binding domain. Expression of the N-terminal/conserved region (CR) 1 or CR3, but not of the CR2 or the C-terminal portion of E1A, inhibited yeast growth. Growth inhibition was relieved by deletion of the genes encoding the yGcn5p, Ngg1p, or Spt7p components of the SAGA transcriptional regulatory complex, but not the Ahc1p component of the related ADA complex, indicating that the N-terminal/CR1 and CR3 regions of E1A target the SAGA complex independently. Expression of the pCAF acetyltransferase, a mammalian homologue of yGcn5p, also suppressed growth inhibition by either portion of E1A. Furthermore, the N-terminal 29 residues and the CR3 portion of E1A interacted independently with yGcn5p and pCAF in vitro. Thus, two separate regions of E1A target the yGcn5p component of the SAGA transcriptional activation complex. A subregion of the N-terminal/CR1 fragment spanning residues 30-69 within CR1 also inhibited yeast growth in a SAGA-dependent fashion. However, this region did not interact with yGcn5p or pCAF, suggesting that it makes a third contact with another SAGA component. Our results provide a new model system to elucidate mechanisms by which E1A and the SAGA complex regulate transcription and growth. PMID:12070146

  1. 11 CFR 300.62 - Non-Federal elections (2 U.S.C. 441i(e)(1)(B)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... elections (2 U.S.C. 441i(e)(1)(B)). A person described in 11 CFR 300.60 may solicit, receive, direct... 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...

  2. 11 CFR 300.61 - Federal elections (2 U.S.C. 441i(e)(1)(A)).

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... elections (2 U.S.C. 441i(e)(1)(A)). No person described in 11 CFR 300.60 shall solicit, receive, direct... any Federal election activity as defined in 11 CFR 100.24, unless the amounts consist of Federal funds... 11 Federal Elections 1 2010-01-01 2010-01-01 false Federal elections (2 U.S.C. 441i(e)(1)(A))....

  3. Enhanced metabolism of halogenated hydrocarbons in transgenic plants containing mammalian cytochrome P450 2E1

    PubMed Central

    Doty, Sharon Lafferty; Shang, Tanya Q.; Wilson, Angela M.; Tangen, Jeff; Westergreen, Aram D.; Newman, Lee A.; Strand, Stuart E.; Gordon, Milton P.

    2000-01-01

    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. PMID:10841534

  4. Epidural blood flow during prostaglandin E1 or trimethaphan induced hypotension.

    PubMed

    Abe, K; Kakiuchi, M; Shimada, Y

    1993-11-01

    To evaluate the effect of prostaglandin E1 (PGE1) or trimethaphan (TMP) induced hypotension on epidural blood flow (EBF) during spinal surgery, EBF was measured using the heat clearance method in 30 patients who underwent postero-lateral interbody fusion under isoflurane anaesthesia. An initial dose of 0.1 microgram.kg-1.min-1 of PGE1 (15 patients), or 10 micrograms.kg-1.min-1 of TMP (15 patients) was administered intravenously after the dural opening and the dose was adjusted to maintain the mean arterial blood pressure (MAP) at about 60 mmHg. The hypotensive drug was discontinued at the completion of the operative procedure. After starting PGE1 or TMP, MAP and rate pressure product (RPP) decreased significantly compared with preinfusion values (P < 0.01), and the degree of hypotension due to PGE1 remained constant until 60 min after its discontinuation. Heart rate (HR) did not change in either group. EBFF did not change during PGE1 infusion whereas in the TMP group, EBF decreased significantly at 30 and 60 min after the start of TMP (preinfusion: 45.9 +/- 13.9 ml/100g/min. 30 min: 32.3 +/- 9.9 ml/100 g/min (P < 0.05). 60 min: 30 +/- 7.5 ml/100 g/min (P < 0.05)). These results suggest that PGE1 may be preferable to TMP for hypotensive anaesthesia in spinal surgery because TMP decreased EBF.

  5. 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

  6. Deficiency of pyruvate dehydrogenase caused by novel and known mutations in the E1alpha subunit.

    PubMed

    Cameron, Jessie M; Levandovskiy, Valeriy; Mackay, Neviana; Tein, Ingrid; Robinson, Brian H

    2004-11-15

    Pyruvate dehydrogenase (PDH)-complex deficiency (OMIM 312170) is a clinically heterogeneous disorder, with phenotypes ranging from fatal lactic acidosis (LA) in the newborn to chronic neurological dysfunction. To date, over 80 different mutations have been identified in the PDHA1 gene in patients with PDH complex deficiency, which are thus thought to contribute to the PDH deficient phenotype. We have identified 14 additional patients with total PDH complex deficiency, all of whom were found to contain mutations within the PDHA1 gene (E(1)alpha subunit). The mutations include both missense mutations and duplications. Eight of these patients had novel mutations, and the remaining had mutations that have been identified previously in PDH complex deficient patients, with residual fibroblast activity ranging from 2.4 to 69% of control values. The nature of these mutations illustrates the variability in phenotype for a given gene defect, with intermittent ataxia being the mildest presentation, Leigh syndrome being the most common and severe neonatal LA the most severe.

  7. [The effect of prostaglandin E1 on body temperature, catecholamines and stress hormones during prolonged surgery].

    PubMed

    Okuda, M; Amano, H; Furuhashi, K; Nakai, Y

    1996-01-01

    The effects of prostaglandin E1 (PGE1) on body temperature, catecholamines and stress hormones were evaluated in 10 patients undergoing elective prolonged surgery over 12 hours. PGE1 (0.03 microgram.kg-1.min-1) was administered in 5 patients and was not administered in 5 patients. Deep skin-surface temperature gradients were 5.1 +/- 2.3 degrees C in PGE1 non-administered group and 0.8 +/- 0.9 degree C in PGE1 administered group (P < 0.05). Pharyngeal-skin surface temperature gradients were 8.8 +/- 2.1 degrees C in PGE1 non-administered group and 1.5 +/- 1.5 degrees C in PGE1 administered group (P < 0.05). There were no significant differences between the two groups in respects to catecholamines, stress hormones, lactate level and blood sugar. PGE1 0.03 microgram.kg-1.min-1 is effective in maintaining peripheral circulation without causing body temperature changes during prolonged surgery.

  8. Prostaglandin E1 decreases the low-density-lipoprotein entry into rabbit arterial wall.

    PubMed Central

    Sinzinger, H.; Virgolini, I.; Lupattelli, G.; Molinari, E.; Gerakakis, A.; Angelberger, P.

    1991-01-01

    1. In 72 male rabbits fed a 1% cholesterol supplemented diet the effect of a 4 weeks daily infusion of prostaglandin E1 (PGE1, 20 micrograms kg-1 min-1 over 2 h) on [125I]-low density lipoprotein (LDL) accumulation (10 microCi; 0.5 mg protein ml-1) was examined versus sham-treatment after removal of the endothelium of the abdominal aorta by a Fogarthy catheter. 2. The uptake of [125I]-LDL was significantly (P less than 0.01) higher in endothelium-free aortic segments (showing the highest peak maximum at around 12 h after 125I-injection) as compared to aortic segments with endothelium intact (showing the lowest uptake of [125I]-LDL with the peak maximum at 48 h, last control time). Segments with the endothelium restored showed a similar LDL-retention curve to segments with endothelium however, being again significantly (P less than 0.01) higher. 3. PGE1-treatment caused reduction in LDL-accumulation, being significantly (P less than 0.001) pronounced in segments without endothelium and in segments with endothelium restored. 4. The findings indicate a beneficial effect of PGE1 in lipid metabolism by decreasing the LDL-influx into the arterial wall in-vivo. PMID:1933127

  9. 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.

  10. 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

  11. 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.

  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. 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.

  14. 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.

  15. Enhanced metabolism of halogenated hydrocarbons in transgenic plants containing mammalian cytochrome P450 2E1.

    PubMed

    Doty, S L; Shang, T Q; Wilson, A M; Tangen, J; Westergreen, A D; Newman, L A; Strand, S E; Gordon, M 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.

  16. Inhibition of human papillomavirus DNA replication by small molecule antagonists of the E1-E2 protein interaction.

    PubMed

    White, Peter W; Titolo, Steve; Brault, Karine; Thauvette, Louise; Pelletier, Alex; Welchner, Ewald; Bourgon, Lise; Doyon, Louise; Ogilvie, William W; Yoakim, Christiane; Cordingley, Michael G; Archambault, Jacques

    2003-07-18

    Human papillomavirus (HPV) DNA replication is initiated by recruitment of the E1 helicase by the E2 protein to the viral origin. Screening of our corporate compound collection with an assay measuring the cooperative binding of E1 and E2 to the origin identified a class of small molecule inhibitors of the protein interaction between E1 and E2. Isothermal titration calorimetry and changes in protein fluorescence showed that the inhibitors bind to the transactivation domain of E2, the region that interacts with E1. These compounds inhibit E2 of the low risk HPV types 6 and 11 but not those of high risk HPV types or of cottontail rabbit papillomavirus. Functional evidence that the transactivation domain is the target of inhibition was obtained by swapping this domain between a sensitive (HPV11) and a resistant (cottontail rabbit papillomavirus) E2 type and by identifying an amino acid substitution, E100A, that increases inhibition by approximately 10-fold. This class of inhibitors was found to antagonize specifically the E1-E2 interaction in vivo and to inhibit HPV DNA replication in transiently transfected cells. These results highlight the potential of the E1-E2 interaction as a small molecule antiviral target.

  17. Adenovirus E1A coding sequences that enable ras and pmt oncogenes to transform cultured primary cells.

    PubMed Central

    Zerler, B; Moran, B; Maruyama, K; Moomaw, J; Grodzicker, T; Ruley, H E

    1986-01-01

    Plasmids expressing partial adenovirus early region 1A (E1A) coding sequences were tested for activities which facilitate in vitro establishment (immortalization) of primary baby rat kidney cells and which enable the T24 Harvey ras-related oncogene and the polyomavirus middle T antigen (pmt) gene to transform primary baby rat kidney cells. E1A cDNAs expressing the 289- and 243-amino acid proteins expressed both E1A transforming functions. Mutant hrA, which encodes a 140-amino acid protein derived from the amino-terminal domain shared by the 289- and 243-amino acid proteins, enabled ras (but not pmt) to transform and facilitated in vitro establishment to a low, but detectable, extent. These studies suggest that E1A functions which collaborate with ras oncogenes and those which facilitate establishment are linked. Furthermore, E1A transforming functions are not associated with activities of the 289-amino acid E1A proteins required for efficient transcriptional activation of viral early region promoters. Images PMID:3022137

  18. 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

  19. E1 and E2 transitions for Fe XVI, Co XVII and Ni XVIII

    NASA Astrophysics Data System (ADS)

    Çelik, G.; Ateş, Ş.

    2016-07-01

    Electric dipole and electric quadrupole transition data for sodium-like iron, cobalt and nickel have been calculated within the weakest bound electron potential model (WBEPM) theory using experimental energy levels and theoretical expectation values of orbital radii corresponding to those energy levels under the assumption of the {LS} coupling scheme. The results obtained from this study provide theoretical transition probability and oscillator strength data requested in many fields of researches, especially astrophysics. The calculated transition data results have been compared with available data in the literature. The present results are consistent with earlier calculations. Some new electric quadrupole transition probability values not existing in the data bases, especially for iron have been obtained using this method.

  20. 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.

  1. 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.

  2. 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

  3. 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

  4. Dechlorination and Decolorization of Organics in Bleach Plant E-1 Effluent by Photochemical Processes

    NASA Astrophysics Data System (ADS)

    Xie, Tianyan

    1994-01-01

    Photochemical study of the dechlorination of four model compounds, 4,5-dichloroguaiacol, 2,4,6-trichlorophenol, 2,3,4,5-tetrachlorophenol, and tetrachloroguaiacol in aqueous solutions under UV radiation was conducted using ArF (193 nm) and KrF (248 nm) excimer laser to explore the response of chlorinated phenolics present in the E_1 effluent from conventional chlorine bleaching of softwood kraft pulp towards photo-oxidation processes. Kinetic study show that the overall dechlorination reaction follow the first order rate law. The factors affecting the dechlorination were investigated. The quantum yield of chloride ion formation was found to be dependent on pH of the reaction mixture, and orignal chlorine content of the compounds. The effect of the substituents on the aromatic ring on the reactivity of the compounds was studied. The mechanism for the dechlorination was proposed involving homolytic photo-dissociation, heterolytic cleavage of carbon-chlorine bonds and substitution reactions of hydroxyl radicals. It was found that the dechlorination under formation to chloride is influenced by the amount of organically bound chlorine in the starting material. Dechlorination reaction favors high pH. Guaiacols more easily undergo dechlorination than phenols. Four fractions of high relative molecular-mass chloro-organics or polychlorinated oxylignin (PCOL) were isolated from an E_1 effluent by combination of ultrafiltration, and purified by repeated precipitation. The fractions were analysed by classical functional group analysis and spectrophotometric methods. The analytical data indicated that the major structural differences between PCOL fractions and kraft lignin preparations are with regard to the content of founctional groups such as carboxyl content, methoxyl and hydroxyl contents. In addition, IR, ^1H and ^{13 }C NMR spectral analyses revealed an almost complete absence of absorption attributable to aromatic structures in PCOLs. These results and others led to the

  5. 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.

  6. Effects of Ca++ and Prostaglandin E1 on Vasopressin Activation of Renal Adenyl Cyclase

    PubMed Central

    Marumo, Fumiaki; Edelman, Isidore S.

    1971-01-01

    Adenyl cyclase activity was assayed in crude homogenates of the renal cortex, medulla, and papilla of the golden hamster. The specific activity (moles C-AMP/unit of time per mg protein of tissue) of the enzyme under basal conditions, was greatest in papilla, somewhat lower in medulla, and least in cortex. On an absolute scale, the sensitivity to vasopressin was greater in the medullary and papillary than in the cortical homogenates. In addition, at concentrations of 0.1-1.0 mm, CaCl2 inhibited the enzyme in the order papilla > medulla > cortex. These results imply the existence of distinct differences in the composition of the adenyl cyclase-receptor complex in various parts of the kidney. We proposed that Ca++ inhibits the core enzyme directly since at the minimally inhibitory concentration (0.1 mm), CaCl2 reduced to an equivalent extent (a) basal activity, (b) the response to graded doses of vasopressin (0.5 to 50.0 mU/ml) and (c) the response to maximal stimulatory concentrations of NaF (10 mm). Prostaglandin E1 (PGE1 = 10−7m) had no effect on either basal adenyl-cyclase activity or the response to 10 mm NaF in medullary and papillary homogenates. 7-Oxa-13-prostynoic acid (10−4m) similarly had no effect under basal conditions or on stimulation with NaF in medullary homogenates. Both fatty acids, however, inhibited the enzymic response to vasopressin, particularly at low concentrations of the peptide. The straight-chain fatty acid, 11-eicosanoic acid (10−7m), was inactive on basal activity or on the response to vasopressin. The possibility that PGE1 modifies the coupling mechanism between the core enzyme and the hormone-specific receptor is discussed. PMID:4329002

  7. Inositol hexakisphosphate inhibits mineralization of MC3T3-E1 osteoblast cultures.

    PubMed

    Addison, William N; McKee, Marc D

    2010-04-01

    Inositol hexakisphosphate (IP6, phytic acid) is an endogenous compound present in mammalian cells and tissues. Differentially phosphorylated forms of inositol are well-documented to have important roles in signal transduction, cell proliferation and differentiation, and IP6 in particular has been suggested to inhibit soft tissue calcification (specifically renal and vascular calcification) by binding extracellularly to calcium oxalate and calcium phosphate crystals. However, the effects of IP6 on bone mineralization are largely unknown. In this study, we used MC3T3-E1 osteoblast cultures to examine the effects of exogenous IP6 on osteoblast function and matrix mineralization. IP6 at physiologic concentrations caused a dose-dependent inhibition of mineralization without affecting cell viability, proliferation or collagen deposition. Osteoblast differentiation markers, including tissue-nonspecific alkaline phosphatase activity, bone sialoprotein and osteocalcin mRNA levels, were not adversely affected by IP6 treatment. On the other hand, IP6 markedly increased protein and mRNA levels of osteopontin, a potent inhibitor of crystal growth and matrix mineralization. Inositol alone (without phosphate), as well as inositol hexakis-sulphate, a compound with a high negative charge similar to IP6, had no effect on mineralization or osteopontin induction. Binding of IP6 to mineral crystals from the osteoblast cultures, as well as to synthetic hydroxyapatite crystals, was confirmed by a colorimetric assay for IP6. In summary, IP6 inhibits mineralization of osteoblast cultures by binding to growing crystals through negatively charged phosphate groups and by induction of inhibitory osteopontin expression. These data suggest that IP6 may regulate physiologic bone mineralization by directly acting extracellularly, and by serving as a specific signal at the cellular level for the regulation of osteopontin gene expression.

  8. Effects of two different programs of modern sports dancing on motor coordination, strength, and speed.

    PubMed

    Uzunovic, Slavoljub; Kostic, Radmila; Zivkovic, Dobrica

    2010-09-01

    This study aimed to determine the effects of two different programs of modern sports dancing on coordination, strength, and speed in 60 beginner-level female dancers, aged 13 and 14 yrs. The subjects were divided into two experimental groups (E1 and E2), each numbering 30 subjects, drawn from local dance clubs. In order to determine motor coordination, strength, and speed, we used 15 measurements. The groups were tested before and after the experimental programs. Both experimental programs lasted for 18 wks, with training sessions twice a week for 60 minutes. The subjects from the E1 group trained according to a new experimental program of disco dance (DD) modern sports dance, and the E2 group trained according to the classic DD program of the same kind for beginner selections. The obtained results were assessed by statistical analysis: a paired-samples t-test and MANCOVA/ANCOVA. The results indicated that following the experimental programs, both groups showed a statistically significant improvement in the evaluated skills, but the changes among the E1 group subjects were more pronounced. The basic assumption of this research was confirmed, that the new experimental DD program has a significant influence on coordination, strength, and speed. In relation to these changes, the application of the new DD program was recommended for beginner dancers. PMID:21120267

  9. Effects of two different programs of modern sports dancing on motor coordination, strength, and speed.

    PubMed

    Uzunovic, Slavoljub; Kostic, Radmila; Zivkovic, Dobrica

    2010-09-01

    This study aimed to determine the effects of two different programs of modern sports dancing on coordination, strength, and speed in 60 beginner-level female dancers, aged 13 and 14 yrs. The subjects were divided into two experimental groups (E1 and E2), each numbering 30 subjects, drawn from local dance clubs. In order to determine motor coordination, strength, and speed, we used 15 measurements. The groups were tested before and after the experimental programs. Both experimental programs lasted for 18 wks, with training sessions twice a week for 60 minutes. The subjects from the E1 group trained according to a new experimental program of disco dance (DD) modern sports dance, and the E2 group trained according to the classic DD program of the same kind for beginner selections. The obtained results were assessed by statistical analysis: a paired-samples t-test and MANCOVA/ANCOVA. The results indicated that following the experimental programs, both groups showed a statistically significant improvement in the evaluated skills, but the changes among the E1 group subjects were more pronounced. The basic assumption of this research was confirmed, that the new experimental DD program has a significant influence on coordination, strength, and speed. In relation to these changes, the application of the new DD program was recommended for beginner dancers.

  10. 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

  11. Induction of brain cytochrome P450 2E1 boosts the locomotor-stimulating effects of ethanol in mice.

    PubMed

    Ledesma, Juan Carlos; Miquel, Marta; Pascual, María; Guerri, Consuelo; Aragon, Carlos M G

    2014-10-01

    In the central nervous system ethanol (EtOH) is metabolized into acetaldehyde by different enzymes. Brain catalase accounts for 60% of the total production of EtOH-derived acetaldehyde, whereas cerebral cytochrome P450 2E1 (CYP 2E1) produces 20% of this metabolite. Acetaldehyde formed by the activity of central catalase has been implicated in some of the neurobehavioral properties of EtOH, yet the contribution of CYP 2E1 to the pharmacological actions of this drug has not been investigated. Here we assessed the possible participation of CYP 2E1 in the behavioral effects of EtOH. Thus, we induced CYP 2E1 activity and expression by exposing mice to chronic acetone intake (1% v/v for 10 days) and examined its consequences on the stimulating and uncoordinating effects of EtOH (0-3.2 g/kg) injected intraperitoneally. Our data showed that 24 h after withdrawal of acetone brain expression and activity of CYP 2E1 was induced. Furthermore, the locomotion produced by EtOH was boosted over the same interval of time. Locomotor stimulation produced by amphetamine or tert-butanol was unchanged by previous treatment with acetone. EtOH-induced motor impairment as evaluated in a Rota-Rod apparatus was unaffected by the preceding exposure to acetone. These results indicate that cerebral CYP 2E1 activity could contribute to the locomotor-stimulating effects of EtOH, and therefore we suggest that centrally produced acetaldehyde might be a possible mediator of some EtOH-induced pharmacological effects.

  12. A non-allergenic Ole e 1-like protein from birch pollen as a tool to design hypoallergenic vaccine candidates.

    PubMed

    Marazuela, Eva G; Hajek, Roswitha; Villalba, Mayte; Barber, Domingo; Breiteneder, Heimo; Rodríguez, Rosalía; Batanero, Eva

    2012-02-01

    Recombinant DNA technology offers several approaches to convert allergens into hypoallergenic derivatives that can represent the basis of novel, safer and more effective forms of allergy vaccines. In this context, we used a new strategy for the design of a hypoallergenic derivative of Ole e 1, the main allergen of olive pollen. By screening a cDNA library from birch pollen, the clone BB18, encoding the birch counterpart of Ole e 1, was identified. In this study, BB18 has been produce in Pichia pastoris as a recombinant protein and immunologically characterized. The well-established non-allergenic properties of BB18 were used to generate a genetic variant of Ole e 1, named OB(55-58), by site-direct mutagenesis of four residues (E(55)V(56)G(57)Y(58)) in an IgE/IgG epitope of Ole e 1 by the corresponding ones in BB18 (SDSE). OB(55-58) was expressed in P. pastoris, purified to homogeneity and analyzed for IgE-reactivity by means of ELISA using sera from olive pollen allergic patients and rat basophil activation assay. T cell reactivity was assayed in a mouse model of Ole e 1 sensitization. The mutant OB(55-58) exhibited an impaired IgE reactivity, but not affected T cell reactivity, compared to wild type rOle e 1. This study emphasizes the usefulness of BB18 as a tool for epitope mapping and for engineering hypoallergenic derivatives of Ole e 1 as vaccine candidates for allergy prevention and treatment.

  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. Radiative rates for E1, E2, M1, and M2 transitions in S-like to F-like tungsten ions (W LIX to W LXVI)

    NASA Astrophysics Data System (ADS)

    Aggarwal, Kanti M.; Keenan, Francis P.

    2016-09-01

    Calculations of energy levels, radiative rates and lifetimes are reported for eight ions of tungsten, i.e. S-like (W LIX) to F-like (W LXVI). A large number of levels have been considered for each ion and extensive configuration interaction has been included among a range of configurations. For the calculations, the general-purpose relativistic atomic structure package (GRASP) has been adopted, and radiative rates (as well as oscillator strengths and line strengths) are listed for all E1, E2, M1, and M2 transitions of the ions. Comparisons have been 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).

  15. 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.

  16. The GST T1 and CYP2E1 genotypes are possible factors causing vinyl chloride induced abnormal liver function.

    PubMed

    Huang, C Y; Huang, K L; Cheng, T J; Wang, J D; Hsieh, L L

    1997-01-01

    Vinyl chloride monomer (VCM) is hepatotoxic as well as carcinogenic in humans. There are reports that exposure to VCM seems to induce abnormal liver function, liver fibrosis, cirrhosis, portal hypertension, and angiosarcoma of the liver. In vivo, VCM is metabolized by cytochrome P450 2E1 (CYP2E1) to form the electrophilic metabolites, chloroethylene oxide (CEO) and chloroacetaldehyde (CAA), which may either cause cell damage or be further metabolized and detoxified by glutathione S-transferases (GSTs). This study investigated whether or not the genotypes CYP2E1, glutathione S-transferase theta (GST T1) and mu (GST M1) correlated with abnormal liver function found in vinyl chloride exposed workers. For this study, 251 workers from five polyvinyl chloride plants were enrolled. The workers were classified into two exposure groups (high and low) and the degree of exposure was determined based on their job titles and airborne VCM concentration. The activity of serum alanine aminotransferase (ALT) was used as the parameter of liver function. The genotypes CYP2E1, GST T1 and GST M1 were determined by polymerase chain reaction and restriction fragment length polymorphism on peripheral white blood cell DNA. Other potential risk factors were also ascertained and the confounding effect was adjusted accordingly. Stratified analyses were used to explore the correlation between the alteration of liver function and the genotypes CYP2E1, GST T1 and GST M1 among the workers exposed to different levels of VCM. The following results were obtained (1) at low VCM exposure, the odds ratio (OR) of positive GST T1 on abnormal ALT was 3.8 (95% CI 1.2-14.5) but the CYP2E1 genotype was not associated with abnormal ALT. (2) At high VCM exposure, a c2c2 CYP2E1 genotype was associated with increased OR on abnormal ALT (OR 5.4, 95% CI 0.7-35.1) and positive GST T1 was significantly associated with decreased OR on abnormal ALT (OR 0.3, 95% CI 0.1-0.9). (3) Multiple linear and logistic regression

  17. 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.

  18. Transition Strength Ratios in the Tetrahedral Candidate ^156Dy

    NASA Astrophysics Data System (ADS)

    Hartley, D. J.; Riedinger, L. L.; Curien, D.; Dudek, J.; Gall, B.; Allmond, J. M.; Beausang, C. W.; Carpenter, M. P.; Chiara, C. J.; Janssens, R. V. F.; Kondev, F. G.; Lauritsen, T.; McCutchan, E. A.; Stefanescu, I.; Zhu, S.; Garrett, P. E.; Kulp, W. D.; Wood, J. L.; Mazurek, K.; Riley, M. A.; Wang, X.; Schunck, N.; Yu, C.-H.; Sharpey-Schafer, J.; Simpson, J.

    2009-10-01

    A new symmetry has been recently proposed where nuclei may stabilize in a tetrahedral (pyramid) shape. One of the consequences of this symmetry is that the transition strength, B(E2), of the inband transitions should approach zero in the ideal case. Thus, one signal of this exotic shape would be a rotational band where the inband E2 transitions are extremely weak or nonexistent. Such bands exist in many of the lowest negative-parity bands in the N 90 nuclei, which is also a predicted ``magic" region for tetrahedral symmetry. A Gammasphere experiment was performed to measure the B(E2)/B(E1) ratios of such a negative-parity band in ^156Dy. The results (which are consistent with the theory) will be presented, as well as a discussion of the proposed follow-up experiment to directly measure the B(E2) rates.

  19. CYP2E1 induction leads to oxidative stress and cytotoxicity in glutathione-depleted cerebellar granule neurons.

    PubMed

    Valencia-Olvera, Ana Carolina; Morán, Julio; Camacho-Carranza, Rafael; Prospéro-García, Oscar; Espinosa-Aguirre, Jesús Javier

    2014-10-01

    Increasing evidence suggests that brain cytochrome P450 (CYP) can contribute to the in situ metabolism of xenobiotics. In the liver, some xenobiotics can be metabolized by CYPs into more reactive products that can damage hepatocytes and induce cell death. In addition, normal CYP activity may produce reactive oxygen species (ROS) that contribute to cell damage through oxidative mechanisms. CYP2E1 is a CYP isoform that can generate ROS leading to cytotoxicity in multiple tissue types. The aim of this study was to determine whether CYP2E1 induction may lead to significant brain cell impairment. Immunological analysis revealed that exposure of primary cerebellar granule neuronal cultures to the CYP inducer isoniazid, increased CYP2E1 expression. In the presence of buthionine sulfoximine, an agent that reduces glutathione levels, isoniazid treatment also resulted in reactive oxygen species (ROS) production, DNA oxidation and cell death. These effects were attenuated by simultaneous exposure to diallyl sulfide, a CYP2E1 inhibitor, or to a mimetic of superoxide dismutase/catalase, (Euka). These results suggest that in cases of reduced antioxidant levels, the induction of brain CYP2E1 could represent a risk of in situ neuronal damage.

  20. Interaction of the Dr1 inhibitory factor with the TATA binding protein is disrupted by adenovirus E1A.

    PubMed Central

    Kraus, V B; Inostroza, J A; Yeung, K; Reinberg, D; Nevins, J R

    1994-01-01

    Past experiments have shown that the adenovirus E1A12S product activates the hsp70 promoter, dependent on the TATA element and dependent on N-terminal E1A sequences. Other experiments have identified a factor termed Dr1 that interacts with and inhibits the transcriptional activity of the TATA-binding protein (TBP). We now find that the E1A12S protein can disrupt the interaction of the Dr1 factor with the TATA-specific TBP factor, allowing the productive interaction of TBP with TFIIA. This E1A-mediated disruption is dependent on N-terminal sequences that are also essential for the TATA-dependent trans-activation of the hsp70 promoter. Moreover, we also find that Dr1 expression in transfected cells can inhibit transcription from the hsp70 promoter and that this can be overcome by coexpression of the wild-type E1A protein, dependent on N-terminal sequences. We conclude that the activation of hsp70 through the TATA element may be mechanistically similar to the activation of the E2 promoter via E2F, in each case involving a release of a transcription factor from an inactive complex. Images PMID:8022773

  1. CYP2E1 induction leads to oxidative stress and cytotoxicity in glutathione-depleted cerebellar granule neurons.

    PubMed

    Valencia-Olvera, Ana Carolina; Morán, Julio; Camacho-Carranza, Rafael; Prospéro-García, Oscar; Espinosa-Aguirre, Jesús Javier

    2014-10-01

    Increasing evidence suggests that brain cytochrome P450 (CYP) can contribute to the in situ metabolism of xenobiotics. In the liver, some xenobiotics can be metabolized by CYPs into more reactive products that can damage hepatocytes and induce cell death. In addition, normal CYP activity may produce reactive oxygen species (ROS) that contribute to cell damage through oxidative mechanisms. CYP2E1 is a CYP isoform that can generate ROS leading to cytotoxicity in multiple tissue types. The aim of this study was to determine whether CYP2E1 induction may lead to significant brain cell impairment. Immunological analysis revealed that exposure of primary cerebellar granule neuronal cultures to the CYP inducer isoniazid, increased CYP2E1 expression. In the presence of buthionine sulfoximine, an agent that reduces glutathione levels, isoniazid treatment also resulted in reactive oxygen species (ROS) production, DNA oxidation and cell death. These effects were attenuated by simultaneous exposure to diallyl sulfide, a CYP2E1 inhibitor, or to a mimetic of superoxide dismutase/catalase, (Euka). These results suggest that in cases of reduced antioxidant levels, the induction of brain CYP2E1 could represent a risk of in situ neuronal damage. PMID:24929095

  2. 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}.

  3. Strength Training and Shoulder Proprioception

    PubMed Central

    Salles, José Inácio; Velasques, Bruna; Cossich, Victor; Nicoliche, Eduardo; Ribeiro, Pedro; Amaral, Marcus Vinicius; Motta, Geraldo

    2015-01-01

    Context: Proprioception is essential to motor control and joint stability during daily and sport activities. Recent studies demonstrated that athletes have better joint position sense (JPS) when compared with controls matched for age, suggesting that physical training could have an effect on proprioception. Objective: To evaluate the result of an 8-week strength-training program on shoulder JPS and to verify whether using training intensities that are the same or divergent for the shoulder's dynamic-stabilizer muscles promote different effects on JPS. Design: Randomized controlled clinical trial. Setting: We evaluated JPS in a research laboratory and conducted training in a gymnasium. Patients or Other Participants: A total of 90 men, right handed and asymptomatic, with no history of any type of injury or shoulder instability. Intervention(s): For 8 weeks, the participants performed the strength-training program 3 sessions per week. We used 4 exercises (bench press, lat pull down, shoulder press, and seated row), with 2 sets each. Main Outcome Measure(s): We measured shoulder JPS acuity by calculating the absolute error. Results: We found an interaction between group and time. To examine the interaction, we conducted two 1-way analyses of variance comparing groups at each time. The groups did not differ at pretraining; however, a difference among groups was noted posttraining. Conclusions: Strength training using exercises at the same intensity produced an improvement in JPS compared with exercises of varying intensity, suggesting that the former resulted in improvements in the sensitivity of muscle spindles and, hence, better neuromuscular control in the shoulder. PMID:25594912

  4. Dechlorination of chlorophenols found in pulp bleach plant E-1 effluents by advanced oxidation processes.

    PubMed

    Wang, Rui; Chen, Chen-Loung; Gratzl, Josef S

    2005-05-01

    Studies were conducted on the response of 2,4,6-trichlorophenol (1), 2,3,4,5-tetrachloro-phenol (2) and 4,5-dichloroguaiacol (3) toward advanced oxidation processes, such as UV-, O2/UV-, H2O2/UV-, O3/UV- and O3-H2O2/UV-photolyses with irradiation of 254 nm photons. The compounds 1-3 are among the chlorophenols found in the Kraft-pulp bleach plant E-1 effluents. The studies were extended to treatment of these compounds with ozonation and O3-H2O2 oxidation systems in alkaline aqueous solution. Except for the O2/UV-photolysis of 1 and H2O2/UV-photolysis of 2, the dechlorination of 1-3 by O2/UV- and H2O2/UV-potolyses were less effective than the corresponding N2UV-potolysis of 1-3. Guaiacol-type chlorophenols were more readily able to undergo dechlorination than non-guaiacol type chlorophenols by N2/UV-, O2/UV- and H2O2/UV-potolyses. In addition, the efficiency for the dechlorination of 1-3 by N2/UV-, O2/UV- and H2O2/UV-potolyses appeared to be dependent upon the inductive and resonance effects of substituents as well as number and position of chlorine substituent in the aromatic ring of the compounds. The dechlorination of 2 by treatment with O3 alone is slightly more effective than the corresponding the O3/UV-photlysis, whereas the dechlorination of 2 by treatment with the combination of O3 and H2O2 was slightly less effective than the corresponding O3-H2O2/UV-photolysis. In contrast, the dechlorination of 3 on treatment with O3 alone was slightly less effective than the corresponding the O3/UV-photolysis, whereas the dechlorination of 3 on treatment with the combination of O3 and H2O2 was slightly more effective than the corresponding the O3-H2O2/UV-photolysis. In the dechlorination of 2 and 3, chemical species derived from ozone and hydrogen peroxide in alkaline solution were dominant reactions in the O3/UV- and O3-H2O2/UV-photolysis systems as in the O3 and O3-H2O2 oxidation systems. Possible dechlorination mechanisms involved were discussed on the basis of

  5. Mapping Lunar global chemical composition from Chang'E-1 IIM data

    NASA Astrophysics Data System (ADS)

    Yan, Bokun; Xiong, Sheng Qing; Wu, Yunzhao; Wang, Zhenchao; Dong, Lina; Gan, Fuping; Yang, Suming; Wang, Runsheng

    2012-07-01

    The global distribution of the chemical composition of the lunar surface is an important factor helping us to understand the formation and evolution of the Moon. In this paper, formulas were established for deriving FeO, TiO2, Al2O3 and MnO abundances from Chang'E-1 (CE-1) Interference Imaging Spectrometer (IIM) data on the basis of the method "color ratio of UV/VIS and NIR/VIS versus VIS reflectance diagram" which was put forward by Lucey and Blewett. Global high-resolution maps (200 m/pixel) of FeO, TiO2, Al2O3 and MnO were produced, and then compared qualitatively with results from Clementine UVVIS, Lunar Prospector (LP) Gamma-Ray Spectrometer (GRS) and Neutron Prospector (NS) data. The abundance ranges of the above four elements are 0-21.0 wt%, 0-9.5 wt%, 5.4-32.1 wt%, and 0.015-0.28 wt% respectively. The abundance range of FeO is consistent with the results from LP-GRS data reported by Gillis et al. (2004), and the abundance range of TiO2 is consistent with the results from LP-NS data reported by Elphic et al. (2002). Relative abundance distributions of FeO and TiO2 from Clementine and IIM data are slightly different from those from LP-GRS and LP-NS data. In map from the LP-GRS data, FeO abundances are the highest at Oceanus Procellarum and Mare Imbrium. However, in the map from CE-1 IIM data they are the highest at Oceanus Procellarum and Mare Tranquillitatis. Although the spatial resolution of these maps is high, caution must be taken when the maps in this paper are used at the crater scale because they suffer from errors owing to topographically induced shading. In future work, a high-accuracy DEM from Lunar Reconnaissance Orbiter Mission Laser Altimeter (LOLA) data coupled with a photometric model can probably be used to resolve this problem.

  6. Prostaglandin E1 alleviates neuropathic pain and neural dysfunction from entrapment neuropathy associated with diabetes mellitus.

    PubMed

    Natsume, Tadahiro; Iwatsuki, Katsuyuki; Nishizuka, Takanobu; Arai, Tetsuya; Yamamoto, Michiro; Hirata, Hitoshi

    2014-10-01

    In this report, we present the results of investigation of the effects of prostaglandin E1 (PGE1) on entrapment neuropathy using a diabetic rat. A total of 60 male Sprague-Dawley rats were used in the study. The model of tibial nerve entrapment neuropathy associated with diabetes mellitus was created by streptozotocin-induced diabetic rats reared in cages with wire grid flooring. Rats were assigned to four groups: nondiabetic (n = 15), untreated diabetic (n = 15), diabetic treated with 30 μg/kg PGE1 (n = 15), and diabetic treated with 100 μg/kg PGE1 (n = 15). Pain tests and electrophysiological tests were performed at 0, 2, and 4 weeks, and assessments of gait, histology, and mRNA expression levels were performed at 4 weeks after initiating the PGE1 administration. In the 30 and 100 μg groups, the mechanical withdrawal thresholds measured by pain tests at 4 weeks (36.2 ± 16.4 g and 31.7 ± 15.3 g, respectively) and the motor conduction velocity (24.0 ± 0.2 m/s and 24.4 ± 0.3 m/s, respectively) were significantly higher than the untreated diabetic group (all P < 0.05) and lower than the nondiabetic group (all P < 0.001). In the gait analysis, the mean intensities in the 30 and 100 μg group (128.0 ± 20.1 a.u. and 109.0 ± 27.8 a.u., respectively) were significantly higher than the untreated diabetic (P < 0.01) and were not significantly different from the nondiabetic group (P = 0.81). Fiber density (P = 0.46) and fiber diameter (P = 0.15) did not show any significant differences. PGE1 significantly decreased nerve growth factor (NGF) mRNA and increased vascular endothelial growth factor (VEGF) mRNA in the tibial nerve (both P < 0.01). In conclusion, neurological deteriorations of diabetic rats were alleviated with PGE1, which is associated with inhibition of NGF and enhancement of VEGF at the entrapment site.

  7. Interaction Studies of Resolvin E1 Analog (RX-10045) with Efflux Transporters

    PubMed Central

    Cholkar, Kishore; Trinh, Hoang M.; Vadlapudi, Aswani Dutt; Wang, Zhiying; Pal, Dhananjay

    2015-01-01

    Abstract Purpose: Screening interactions of a resolvin E1 analog (RX-10045) with efflux transporters (P-glycoprotein [P-gp], multidrug resistance-associated protein [MRP2], and breast cancer-resistant protein [BCRP]). Methods: Madin-Darby canine kidney cells transfected with P-gp, MRP2, and BCRP genes were selected for this study. [3H]-Digoxin, [3H]-vinblastine and [3H]-abacavir were selected as model substrates for P-gp, MRP2, and BCRP. Uptake and permeability studies across cell monolayer in both apical to basal (AP–BL) and BL–AP of these substrates were conducted in the presence of specific efflux pump inhibitors and RX-10045. Cell viability studies were conducted with increasing concentrations of RX-10045. Results: Uptake studies showed a higher accumulation in the presence of inhibitors (GF120918 and ketoconazole for P-gp; MK571 for MRP2; and β-estradiol for BCRP) as well as RX-10045. Similarly, dose-dependent inhibition studies demonstrated higher accumulation of various substrates ([3H]-digoxin, [3H]-vinblastine, and [3H]-abacavir) in the presence of RX-10045. IC50 values of dose-dependent inhibition of RX-10045 for P-gp, MRP2, and BCRP were 239±11.2, 291±79.2, and 300±42 μM, respectively. Cell viability assay indicated no apparent toxicity up to 350 μM concentration. Enhanced permeability for model substrates was observed in the presence of RX-10045. Uptake studies in human corneal epithelial cells suggest that RX-10045 is a strong inhibitor of organic cation transporter-1 (OCT-1). Conclusions: In summary, the resolvin analog (RX-10045) was identified as a substrate/inhibitor for efflux transporters (MRP2 and BCRP). Also, RX-10045 appears to be a strong inhibitor/substrate of OCT-1. Novel formulation strategies such as nanoparticles, nanomicelles, and liposomes for circumventing efflux barriers and delivering higher drug concentrations leading to a higher therapeutic efficacy may be employed. PMID:25844889

  8. Polyphosphates inhibit extracellular matrix mineralization in MC3T3-E1 osteoblast cultures.

    PubMed

    Hoac, Betty; Kiffer-Moreira, Tina; Millán, José Luis; McKee, Marc D

    2013-04-01

    Studies on various compounds of inorganic phosphate, as well as on organic phosphate added by post-translational phosphorylation of proteins, all demonstrate a central role for phosphate in biomineralization processes. Inorganic polyphosphates are chains of orthophosphates linked by phosphoanhydride bonds that can be up to hundreds of orthophosphates in length. The role of polyphosphates in mammalian systems, where they are ubiquitous in cells, tissues and bodily fluids, and are at particularly high levels in osteoblasts, is not well understood. In cell-free systems, polyphosphates inhibit hydroxyapatite nucleation, crystal formation and growth, and solubility. In animal studies, polyphosphate injections inhibit induced ectopic calcification. While recent work has proposed an integrated view of polyphosphate function in bone, little experimental data for bone are available. Here we demonstrate in osteoblast cultures producing an abundant collagenous matrix that normally show robust mineralization, that two polyphosphates (PolyP5 and PolyP65, polyphosphates of 5 and 65 phosphate residues in length) are potent mineralization inhibitors. Twelve-day MC3T3-E1 osteoblast cultures with added ascorbic acid (for collagen matrix assembly) and β-glycerophosphate (a source of phosphate for mineralization) were treated with either PolyP5 or PolyP65. Von Kossa staining and calcium quantification revealed that mineralization was inhibited in a dose-dependent manner by both polyphosphates, with complete mineralization inhibition at 10μM. Cell proliferation and collagen assembly were unaffected by polyphosphate treatment, indicating that polyphosphate inhibition of mineralization results not from cell and matrix effects but from direct inhibition of mineralization. This was confirmed by showing that PolyP5 and PolyP65 bound to synthetic hydroxyapatite in a concentration-dependent manner. Tissue-nonspecific alkaline phosphatase (TNAP, ALPL) efficiently hydrolyzed the two PolyPs as

  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. Photon Strength Functions below GDER Maximum: Present Status and Outlook

    SciTech Connect

    Krticka, Milan

    2009-01-28

    Existing experimental data on {gamma} decay of energy levels in medium-weight and heavy nuclei at excitation energies well above the pairing gap indicate that the role of nuclear structure effects is small and that this decay is largely governed by the extreme statistical model. Specifically, in line with the validity of Brink hypothesis and the paradigm of photon strength, the average properties of the {gamma} decay can be described by a set of photon strength functions (PSFs) for individual multipolarities and by the level density function. PSFs are directly related to the photoabsorption cross sections {sigma}{sub {gamma}}{sub abs}. It is well known that in the case of E1 radiation this cross section is dominated by the giant dipole electric resonance. However the size of {sigma}{sub {gamma}}{sub abs.} and its {gamma}-ray energy dependence at energies below the neutron threshold are known rather poorly. In the case of higher multipolarities (M1, E2,...) the present knowledge about their roles is much worse. Available information on E1 and M1 PSFs obtained from various kinds of experiments in the {gamma}-ray energy region of interest will be discussed within the widely used models. It will be shown that none of them seems to be able to describe PSFs reasonably for a broad range of nuclei. Emphasis will be also laid on the properties of the scissors mode and other possible resonance-like structures in PSFs at energies below about 10 MeV. Perspectives of further studies of PSFs will be outlined.

  11. 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.

  12. Sulfonate groups grafted on Ti6Al4V favor MC3T3-E1 cell performance in serum free medium conditions.

    PubMed

    Felgueiras, Helena; Migonney, Véronique

    2014-06-01

    Ten years ago, we synthesized "bioactive model polymers" bearing sulfonate groups and proposed a mechanism of their modulation effect at different steps of the cell response. Then, we set up the grafting of polymers bearing sulfonate on Ti6Al4V surfaces by a grafting "from" technique making sure of the creation of covalent bonds between the grafted polymer and the Ti6Al4V surface. We have checked and confirmed the positive effect of grafted sulfonate groups on the osteoblastic cell response in vivo and in vitro but we did not elucidate the mechanism. The aim of this basic work consists first in investigating the role of sulfonate groups in the presence and in the absence of proteins at early stages of the osteointegration process on poly(sodium styrene sulfonate) poly(NaSS) grafted and ungrafted Ti6Al4V surfaces, in vitro. To understand the role of poly(NaSS) grafted chains on osteoblast-like cell response and to confirm/elucidate the importance of fetal bovine serum (FBS) proteins in the culture medium, MC3T3-E1 cells were seeded onto poly(NaSS) grafted and non-grafted Ti6Al4V surfaces. Cultures were carried out in a complete (10% FBS) and in a non-complete medium (without FBS). Cell viability assay, cell attachment number and cell adhesion strength were followed up to 3days of culture. The presence of proteins enhanced cell growth and development whatever the surface and the presence of sulfonate groups enhanced the cell attachment even in the absence of proteins, which suggests and confirms that the sulfonate groups can modify the activity of cells such as the secretion of binding proteins. Statistical differences were found in the attachment strength tests on poly(NaSS) grafted and ungrafted surfaces and showed that the sulfonate groups play an important role in the cell resistance to shear stress. PMID:24863216

  13. Green tea polyphenol (-)-epigallocatechin gallate suppressed the differentiation of murine osteoblastic MC3T3-E1 cells.

    PubMed

    Kamon, Masayoshi; Zhao, Ran; Sakamoto, Kazuichi

    2009-12-16

    Recently, various physiological effects of the tea polyphenol catechin for alleviating diseases such as cancer, arteriosclerosis, hyperlipidaemia and osteoporosis have been reported. However, the physiological effect of catechin on bone metabolism remains unclear. We examined the physiological effect of EGCG [(-)-epigallocatechin-3-gallate], which is the main component of green tea catechin, on osteoblast development using the precursor cell line of osteoblasts, MC3T3-E1, and co-culture of the osteoblasts from mouse newborn calvaria and mouse bone marrow cells. Although EGCG did not affect the viability and proliferation of MC3T3-E1 cells, EGCG inhibited the osteoblast differentiation. Furthermore, EGCG did not affect the mineralization of differentiated MC3T3-E1 cells, and reduced osteoclast formation in co-culture. These results suggest that EGCG can effectively suppress bone resorption, and can be used as an effective medicine in the treatment of the symptoms of osteoporosis.

  14. Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphisms of the CYP2E1 Gene.

    PubMed

    Suhda, Saihas; Paramita, Dewi Kartikawati; Fachiroh, Jajah

    2016-01-01

    Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP). PMID:27509930

  15. Tetra Primer ARMS PCR Optimization to Detect Single Nucleotide Polymorphisms of the CYP2E1 Gene.

    PubMed

    Suhda, Saihas; Paramita, Dewi Kartikawati; Fachiroh, Jajah

    2016-01-01

    Single nucleotide polymorphism (SNP) detection has been used extensively for genetic association studies of diseases including cancer. For mass, yet accurate and more economic SNP detection we have optimized tetra primer amplification refractory mutation system polymerase chain reaction (ARMS PCR) to detect three SNPs in the cytochrome P450 2E1 (CYP2E1) gene locus; i.e. rs3813865, rs2070672 and rs3813867. The optimization system strategies used were (1) designing inner and outer primers; (2) determining of their optimum primer concentration ratios; and (3) determining of the optimum PCR annealing temperature. The tetra primer ARMS PCR result could be directly observed using agarose gel electrophoresis. The method succesfully determined three SNPs in CYP2E1 locus, the results being consistent with validation using DNA sequencing and restriction fragment length polymorphisms (RFLP).

  16. Degradation of low molecular weight volatile organic compounds by plants genetically modified with mammalian cytochrome P450 2E1.

    PubMed

    James, C Andrew; Xin, Gang; Doty, Sharon L; Strand, Stuart E

    2008-01-01

    Cytochrome P450 2E1 (CYP2E1) is a key enzyme in the mammalian metabolism of several low molecular weight volatile organic compounds (VOCs), such as trichloroethylene (TCE), vinyl chloride (VC), carbon tetrachloride (CT), benzene, chloroform, and bromodichloromethane (BDCM), which are all common environmental pollutants that pose risks to human health. We have developed a transgenic tobacco (Nicotiana tabacum cv. Xanthii) that expresses CYP2E1 with increased activity toward TCE and ethylene dibromide. In experiments with tobacco plant cuttings exposed to VOCs in small hydroponic vessels, the transgenic tobacco had greatly increased rates of removal of TCE, VC, CT, benzene, toluene, chloroform, and BDCM, compared to wild-type or vector control tobacco, but not of perchloroethylene or 1,1,1-trichloroethane.

  17. Inhibition of chlorzoxazone metabolism, a clinical probe for CYP2E1, by a single ingestion of watercress.

    PubMed

    Leclercq, I; Desager, J P; Horsmans, Y

    1998-08-01

    To investigate the effect of watercress on the metabolism of chlorzoxazone, an in vivo probe for CYP2E1, the oral pharmacokinetics of chlorzoxazone was studied in 10 healthy volunteers before and after a single ingestion of a watercress homogenate (50 gm). A third chlorzoxazone pharmacokinetic study was performed after a 1-week treatment with isoniazid (300 mg/day), a well-known CYP2E1 inhibitor. Ingestion of watercress or isoniazid did not affect the oral absorption of chlorzoxazone. The area under the chlorzoxazone plasma concentration-time curve was significantly increased by 56% (p < 0.05) after watercress ingestion and by 135% (p < 0.001) with isoniazid treatment. Similarly, chlorzoxazone elimination half-life was prolonged after watercress (53%; p < 0.05) and isoniazid (104%; p < 0.01) administration. These results show that a single ingestion of watercress inhibits the hydroxylation of chlorzoxazone, an in vivo probe for CYP2E1.

  18. The strength of Miranda's lithosphere

    NASA Technical Reports Server (NTRS)

    Pappalardo, Robert; Greeley, Ronald

    1991-01-01

    In attempting to understand the endogenic processes which have shaped the surface of an icy satellite, it is desirable to quantify the failure strength of the satellite's lithosphere. In a crust that is fractured on a large scale, frictional sliding along pre-existing fractures occurs in response to lower differential stresses than required to initiate fracture of pristine rock, thus governing failure of a brittle lithosphere. Failure is predicted along favorably oriented fracture planes; if fractures of all orientations are assumed to be present in the crust (as is expected of a heavily cratered lithosphere), frictional failure relations are directly applicable. The Coulomb criterion predicts that the shear stress (sigma sub t) and normal stress (sigma sub n) components on a fracture plane at failure are related as sigma sub t = mu-sigma sub n + S sub o, where S sub o is the cohesion and mu is the coefficient of friction. At moderate to high pressures, the frictional sliding strength of most materials is found to be sigma sub t = 0.85 sigma sub n.

  19. Strength evaluation of socket joints

    NASA Technical Reports Server (NTRS)

    Rash, Larry C.

    1994-01-01

    This report documents the development of a set of equations that can be used to provide a relatively simple solution for identifying the strength of socket joints and for most cases avoid the need of more lengthy analyses. The analytical approach was verified by comparison of the contact load distributions to results obtained from a finite element analysis. The contacting surfaces for the specific joint in this analysis are in the shape of frustrums of a cone and are representative of the tapered surfaces in the socket-type joints used to join segments of model support systems for wind tunnels. The results are in the form of equations that can be used to determine the contact loads and stresses in the joint from the given geometry and externally applied loads. Equations were determined to define the bending moments and stresses along the length of the joints based on strength and materials principles. The results have also been programmed for a personal computer and a copy of the program is included.

  20. Asteroid airburst altitude vs. strength

    NASA Astrophysics Data System (ADS)

    Robertson, Darrel; Wheeler, Lorien; Mathias, Donovan

    2016-10-01

    Small NEO asteroids (<Ø140m) may not be a threat on a national or global level but can still cause a significant amount of local damage as demonstrated by the Chelyabinsk event where there was over $33 million worth of damage (1 billion roubles) and 1500 were injured, mostly due to broken glass. The ground damage from a small asteroid depends strongly on the altitude at which they "burst" where most of the energy is deposited in the atmosphere. The ability to accurately predict ground damage is useful in determining appropriate evacuation or shelter plans and emergency management.Strong asteroids, such as a monolithic boulder, fail and create peak energy deposition close to the altitude at which ram dynamic pressure exceeds the material cohesive strength. Weaker asteroids, such as a rubble pile, structurally fail at higher altitude, but it requires the increased aerodynamic pressure at lower altitude to disrupt and disperse the rubble. Consequently the resulting airbursts have a peak energy deposition at similar altitudes.In this study hydrocode simulations of the entry and break-up of small asteroids were performed to examine the effect of strength, size, composition, entry angle, and speed on the resulting airburst. This presentation will show movies of the simulations, the results of peak burst height, and the comparison to semi-analytical models.

  1. 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.

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

    PubMed Central

    Ye, Qinyuan; Lian, Fuzhi; Chavez, Pollyanna R.G.; Chung, Jayong; Ling, Wenhua; Qin, Hua; Seitz, Helmut K.

    2012-01-01

    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 ethanol liquid diet or a non-ethanol liquid diet, with or without CMZ for one and ten months. A single intraperitoneal injection of diethylnitrosamine (DEN, 20 mg/kg) was given to initiate hepatic carcinogenesis. CYP2E1 expression, inflammatory proteins, cell proliferation, protein-bound 4-HNE, etheno-DNA adducts, 8-hydroxy-2'-deoxyguanosine (8-OHdG), retinoid concentrations, and hepatic carcinogenesis were examined. Ethanol feeding for 1 month with DEN resulted in significantly increased hepatic CYP2E1 levels and increased nuclear accumulation of NF-κB protein and TNF-α expression, which were associated with increased cyclin D1 expression and p-GST positive altered hepatic foci. All of these changes induced by ethanol feeding were significantly inhibited by the one month CMZ treatment. At 10-months of treatment, hepatocellular adenomas were detected in ethanol-fed rats only, but neither in control rats nor in animals receiving ethanol and CMZ. The 8-OHdG formation was found to be significantly increased in ethanol fed animals and normalized with CMZ treatment. In addition, alcohol-reduced hepatic retinol and retinoic acid concentrations were restored by CMZ treatment to normal levels in the rats at 10 months of treatment. These data demonstrate that the inhibition of ethanol-induced CYP2E1 as a key pathogenic factor can counteract the tumor-promoting action of ethanol by decreasing TNF-α expression, NF-κB activation, and oxidative DNA damage as well as restoring normal hepatic levels of retinoic acid in DEN-treated rats. PMID:23543859

  3. Effect of the CYP2E1 genotype on vinyl chloride monomer-induced liver fibrosis among polyvinyl chloride workers.

    PubMed

    Hsieh, Hui-I; Chen, Pau-Chung; Wong, Ruey-Hong; Wang, Jung-Der; Yang, Pei-Ming; Cheng, Tsun-Jen

    2007-09-24

    Although a relationship between vinyl chloride monomer (VCM) and liver cirrhosis has been reported, the underlying mechanisms are not clear. Cytochrome P450 2E1 (CYP2E1), aldehyde dehydrogenase 2 (ALDH2) and glutathione S-transferase theta 1 (GSTT1) enzymes are involved in activation and detoxification of VCM, and thus may be important determinants of interindividual susceptibility to VCM-induced liver damage, including liver cirrhosis. The objective of this study was to evaluate if metabolizing genetic polymorphisms could modify individual susceptibility to liver fibrosis of the VCM exposure. CYP2E1, ALDH2, and GSTT1 polymorphisms were determined by the PCR-RFLP method among 320 workers who were employed in five polyvinyl chloride manufacturing plants. Cumulative VCM exposure levels for study subjects were calculated using a job exposure matrix model. Thirteen workers were diagnosed as having liver fibrosis by using ultrasonography. We observed a dose-response trend between VCM exposure and liver fibrosis. Regarding the results on genetic polymorphisms, CYP2E1 c2c2 genotype showed a significant increase in the risk of liver fibrosis as compared to those with CYP2E1 c1c1 or c1c2 genotypes. No differences were observed between GSTT1 and ALDH2 genotypes and liver fibrosis. In summary, our result suggests that genetic polymorphism in CYP2E1 may be responsible for individual differences in susceptibility to liver fibrosis with regard to chronic VCM exposure. Thus, polymorphism analysis of metabolizing enzymes might be useful in the risk assessment of liver damage in workers with VCM exposure.

  4. A DNA element that regulates expression of an endogenous retrovirus during F9 cell differentiation is E1A dependent.

    PubMed Central

    Lamb, B T; Satyamoorthy, K; Solter, D; Basu, A; Xu, M Q; Weinmann, R; Howe, C C

    1992-01-01

    The retinoic acid-induced differentiation of F9 cells into parietal endoderm-like cells activates transcription of the endogenous mouse retrovirus, the intracisternal A-particle (IAP). To investigate the elements that control IAP gene differentiation-specific expression, we used methylation interference, Southwestern (DNA-protein), and transient-transfection assays and identified the IAP-proximal enhancer (IPE) element that directs differentiation-specific expression. We find that the IPE is inactive in undifferentiated F9 cells and active in differentiated parietal endoderm-like PYS-2 cells. Three proteins of 40, 60, and 68 kDa bind to the sequence GAGTAGAC located between nucleotides -53 and -47 within the IPE. The 40- and 68-kDa proteins from both the undifferentiated and differentiated cells exhibit similar DNA-binding activities. However, the 60-kDa protein from differentiated cells has greater binding activity than that from undifferentiated cells, suggesting a role for this protein in F9 differentiation-specific expression of the IAP gene. The IAP gene is negatively regulated by the adenovirus E1A proteins, and the E1A sequence responsible for repression is located at the N terminus, between amino acids 2 and 67. The DNA sequence that is the target of E1A repression also maps to the IPE element. Colocalization of the differentiation-specific and E1A-sensitive elements to the same protein-binding site within the IPE suggests that the E1A-like activity functions in F9 cells to repress IAP gene expression. Activation of the IAP gene may result when the E1A-like activity is lost or inactivated during F9 cell differentiation, followed by binding of the 60-kDa positive regulatory protein to the enhancer element. Images PMID:1406664

  5. Cytochrome P450-2E1 promotes aging-related hepatic steatosis, apoptosis and fibrosis through increased nitroxidative stress.

    PubMed

    Abdelmegeed, Mohamed A; Choi, Youngshim; Ha, Seung-Kwon; Song, Byoung-Joon

    2016-02-01

    The role of ethanol-inducible cytochrome P450-2E1 (CYP2E1) in promoting aging-dependent hepatic disease is unknown and thus was investigated in this study. Young (7 weeks) and aged female (16 months old) wild-type (WT) and Cyp2e1-null mice were used in this study to evaluate age-dependent changes in liver histology, steatosis, apoptosis, fibrosis and many nitroxidative stress parameters. Liver histology showed that aged WT mice exhibited markedly elevated hepatocyte vacuolation, ballooning degeneration, and inflammatory cell infiltration compared to all other groups. These changes were accompanied with significantly higher hepatic triglyceride and serum cholesterol in aged WT mice although serum ALT and insulin resistance were not significantly altered. Aged WT mice showed the highest rates of hepatocyte apoptosis and hepatic fibrosis. Further, the highest levels of hepatic hydrogen peroxide, lipid peroxidation, protein carbonylation, nitration, and oxidative DNA damage were observed in aged WT mice. These increases in the aged WT mice were accompanied by increased levels of mitochondrial nitroxidative stress and alteration of mitochondrial complex III and IV proteins in aged WT mice, although hepatic ATP levels seems to be unchanged. In contrast, the aging-related nitroxidative changes were very low in aged Cyp2e1-null mice. These results suggest that CYP2E1 is important in causing aging-dependent hepatic steatosis, apoptosis and fibrosis possibly through increasing nitroxidative stress and that CYP2E1 could be a potential target for translational research in preventing aging-related liver disease. PMID:26703967

  6. Incomplete LPS Core-Specific Felix01-Like Virus vB_EcoM_VpaE1

    PubMed Central

    Šimoliūnas, Eugenijus; Vilkaitytė, Monika; Kaliniene, Laura; Zajančkauskaitė, Aurelija; Kaupinis, Algirdas; Staniulis, Juozas; Valius, Mindaugas; Meškys, Rolandas; Truncaitė, Lidija

    2015-01-01

    Bacteriophages represent a valuable source for studying the mechanisms underlying virus-host interactions. A better understanding of the host-specificity of viruses at the molecular level can promote various phage applications, including bacterial diagnostics, antimicrobial therapeutics, and improve methods in molecular biology. In this study, we describe the isolation and characterization of a novel coliphage, vB_EcoM_VpaE1, which has different host specificity than its relatives. Morphology studies, coupled with the results of genomic and proteomic analyses, indicate that vB_EcoM_VpaE1 belongs to the newly proposed genus Felix01likevirus in the family Myoviridae. The genus Felix01likevirus comprises a group of highly similar phages that infect O-antigen-expressing Salmonella and Escherichia coli (E. coli) strains. Phage vB_EcoM_VpaE1 differs from the rest of Felix01-like viruses, since it infects O-antigen-deficient E. coli strains with an incomplete core lipopolysaccharide (LPS). We show that vB_EcoM_VpaE1 can infect mutants of E. coli that contain various truncations in their LPS, and can even recognize LPS that is truncated down to the inner-core oligosaccharide, showing potential for the control of rough E. coli strains, which usually emerge as resistant mutants upon infection by O-Ag-specific phages. Furthermore, VpaE1 can replicate in a wide temperature range from 9 to 49 °C, suggesting that this virus is well adapted to harsh environmental conditions. Since the structural proteins of such phages tend to be rather robust, the receptor-recognizing proteins of VpaE1 are an attractive tool for application in glycan analysis, bacterial diagnostics and antimicrobial therapeutics. PMID:26633460

  7. RsaI polymorphism at the cytochrome P4502E1 locus and risk of hepatocellular carcinoma.

    PubMed Central

    Ladero, J M; Agúndez, J A; Rodríguez-Lescure, A; Diaz-Rubio, M; Benítez, J

    1996-01-01

    BACKGROUND: CYP2E1, the coding gene for the ethanol inducible cytochrome P4502E1, is polymorphic at the RsaI restriction site in the 5' flanking region. The mutant allele c2 has a higher transcriptional activity than the wild-type gene c1. P4502E1 catalyses the activation of several environmental carcinogens at a rate that is increased, if only moderately, by longterm ethanol intake. AIMS: To establish the distribution of CYP2E1 RsaI polymorphism in patients with hepatocellular carcinoma and to evaluate its possible role in the multifactorial pathogenesis of this tumour. SUBJECTS: 101 (84 males) patients with hepatocellular carcinoma and 178 (128 males) healthy controls of the same ethnic (white) and Spanish origin. METHODS: After extraction of DNA from white blood cells, alleles c1 and c2 of CYP2E1 were identified by restriction fragment length polymorphism (RFLP) with endonuclease RsaI. RESULTS: Homozygous c1c1: 90 patients and 169 controls; heterozygous c1c2: 11 and 9; homozygous c2c2: none (non-significant difference). C2 allele frequencies: 0.055 in patients, 0.025 in controls (non-significant difference) and 0.108 in the 37 patients who had drunk more than 50 g of ethanol/day (p = 0.0035, odds ratio versus controls: 4.67; 95% confidence limits 1.57 to 13.81). CONCLUSION: The carrier state of one copy of the c2 CYP2E1 gene increases the risk of hepatoma in previously regular ethanol users with chronic liver disease. PMID:8977352

  8. Occupational Toluene Exposure Induces Cytochrome P450 2E1 mRNA Expression in Peripheral Lymphocytes

    PubMed Central

    Mendoza-Cantú, Ania; Castorena-Torres, Fabiola; de León, Mario Bermúdez; Cisneros, Bulmaro; López-Carrillo, Lizbeth; Rojas-García, Aurora E.; Aguilar-Salinas, Alberto; Manno, Maurizio; Albores, Arnulfo

    2006-01-01

    Print workers are exposed to organic solvents, of which the systemic toxicant toluene is a main component. Toluene induces expression of cytochrome P450 2E1 (CYP2E1), an enzyme involved in its own metabolism and that of other protoxicants, including some procarcinogens. Therefore, we investigated the association between toluene exposure and the CYP2E1 response, as assessed by mRNA content in peripheral lymphocytes or the 6-hydroxychlorzoxazone (6OH-CHZ)/chlorzoxazone (CHZ) quotient (known as CHZ metabolic ratio) in plasma, and the role of genotype (5′-flanking region RsaI/PstI polymorphic sites) in 97 male print workers. The geometric mean (GM) of toluene concentration in the air was 52.80 ppm (10–760 ppm); 54% of the study participants were exposed to toluene concentrations that exceeded the maximum permissible exposure level (MPEL). The GM of urinary hippuric acid at the end of a work shift (0.041 g/g creatinine) was elevated relative to that before the shift (0.027 g/g creatinine; p < 0.05). The GM of the CHZ metabolic ratio was 0.33 (0–9.3), with 40% of the subjects having ratios below the GM. However, the average CYP2E1 mRNA level in peripheral lymphocytes was 1.07 (0.30–3.08), and CYP2E1 mRNA levels within subjects correlated with the toluene exposure ratio (environmental toluene concentration:urinary hippuric acid concentration) (p = 0.014). Genotype did not alter the association between the toluene exposure ratio and mRNA content. In summary, with further validation, CYP2E1 mRNA content in peripheral lymphocytes could be a sensitive and noninvasive biomarker for the continuous monitoring of toluene effects in exposed persons. PMID:16581535

  9. 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.

  10. 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.

  11. 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…

  12. 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.

  13. Ultrafast carrier relaxation through Auger recombination in the topological insulator B i1.5S b0.5T e1.7S e1.3

    NASA Astrophysics Data System (ADS)

    Onishi, Yoshito; Ren, Zhi; Segawa, Kouji; Kaszub, Wawrzyniec; Lorenc, Maciej; Ando, Yoichi; Tanaka, Koichiro

    2015-02-01

    Ultrafast carrier dynamics have great significance for our understanding of the transport properties of the surface state in topological insulator (TI) materials. We report midinfrared pump-probe measurements on the intrinsic TI material B i1.5S b0.5T e1.7S e1.3 and show that the change in photoinduced reflectivity can be decomposed into a fast negative part and a slow positive part. Calculations of the dielectric function made at various carrier temperatures and densities reveal that the fast negative component corresponds to the disappearance of the phase-space filling effect due to hot carriers around the probe energy and the decay component corresponds to the recombination of carriers near the band edge. The ratio of the fast negative component to the slow positive component is larger in the excitations conducted at the higher carrier densities, which suggests that the carrier temperature increases through Auger recombination. A qualitative analysis using rate equations reinforces this assumption, so we conclude that Auger recombination is the main cause of the population relaxation at carrier densities higher than 1018c m-3 and that we determined the Auger coefficient for B i1.5S b0.5T e1.7S e1.3 as C =0.4 ×10-26c m6/s .

  14. The new ENSEMBLES E1 mitigation scenario for future climate simulations

    NASA Astrophysics Data System (ADS)

    Royer, J.-F.; Lowe, J.; Johns, T.; van Vuuren, D.; Stehfest, E.; Denoblet-Ducoudré, N.; Boucher, O.; Rognerud, B.; Huebener, H.

    2009-04-01

    that equivalent to 450-ppm of CO2 concentration. This new stabilisation scenario (E1) has been developed with the latest version of the integrated impact assessment model IMAGE at Netherlands Environmental Assessment Agency (MNP), with improvements in the land use representation of the A1B scenario. Though the coupled climate models are directly forced by the concentrations of the well-mixed greenhouse gases (CO2, CH4, N2O, CFCs and other minor species) produced by the IMAGE model, it has however been necessary to make further adaptations, by using additional modelling tools, in order to produce the other forcing data needed by the GCMs. The 3-dimensional atmospheric fields for anthropogenic sulphate aerosols concentrations have been computed by a chemistry-transport model using the emissions from the IMAGE scenarios. The ozone concentrations have been computed by the chemistry-transport model from the University of Oslo. The land use maps over the period 1850-2000 have been derived from the LUCID project based on a combination of the crop dataset of Ramankutty and Foley and pasture from the HYDE dataset. They have been then merged with the land-use maps produced by the IMAGE scenarios in order to derive adapted land-use maps over the 21st century. The different forcings and some results of the "stream 2" ENSEMBLES scenarios are described and illustrated in this presentation.

  15. Structure, MC3T3-E1 cell response, and osseointegration of macroporous titanium implants covered by a bioactive microarc oxidation coating with microporous structure.

    PubMed

    Zhou, Rui; Wei, Daqing; Cheng, Su; Feng, Wei; Du, Qing; Yang, Haoyue; Li, Baoqiang; Wang, Yaming; Jia, Dechang; Zhou, Yu

    2014-04-01

    Macroporous Ti with macropores of 50-400 μm size is prepared by sintering Ti microbeads with different diameters of 100, 200, 400, and 600 μm. Bioactive microarc oxidation (MAO) coatings with micropores of 2-5 μm size are prepared on the macroporous Ti. The MAO coatings are composed of a few TiO2 nanocrystals and lots of amorphous phases with Si, Ca, Ti, Na, and O elements. Compared to compact Ti, the MC3T3-E1 cell attachment is prolonged on macroporous Ti without and with MAO coatings; however, the cell proliferation number increases. These results are contributed to the effects of the space structure of macroporous Ti and the surface chemical feature and element dissolution of the MAO coatings during the cell culture. Macroporous Ti both without and with MAO coatings does not cause any adverse effects in vivo. The new bone grows well into the macropores and micropores of macroporous Ti with MAO coatings, showing good mechanical properties in vivo compared to Ti, MAO-treated Ti, and macroporous Ti because of its excellent osseointegration. Moreover, the MAO coatings not only show a high interface bonding strength with new bones but also connect well with macroporous Ti. Furthermore, the pushing out force for macroporous Ti with MAO coatings increases significantly with increasing microbead diameter. PMID:24579697

  16. Analysis of the selective advantage conferred by a C-E1 fusion protein synthesized by rubella virus DI RNAs

    SciTech Connect

    Claus, Claudia; Tzeng, W.-P.; Liebert, Uwe Gerd; Frey, Teryl K.

    2007-12-05

    During serial passaging of rubella virus (RUB) in cell culture, the dominant species of defective-interfering RNA (DI) generated contains an in-frame deletion between the capsid protein (C) gene and E1 glycoprotein gene resulting in production of a C-E1 fusion protein that is necessary for the maintenance of the DI [Tzeng, W.P., Frey, T.K. (2006). C-E1 fusion protein synthesized by rubella virus DI RNAs maintained during serial passage. Virology 356 198-207.]. A BHK cell line stably expressing the RUB structural proteins was established which was used to package DIs into virus particles following transfection with in vitro transcripts from DI infectious cDNA constructs. Packaging of a DI encoding an in-frame C-GFP-E1 reporter fusion protein corresponding to the C-E1 fusion protein expressed in a native DI was only marginally more efficient than packaging of a DI encoding GFP, indicating that the C-E1 fusion protein did not function by enhancing packaging. However, infection with the DI encoding the C-GFP-E1 fusion protein (in the absence of wt RUB helper virus) resulted in formation of clusters of GFP-positive cells and the percentage of GFP-positive cells in the culture following infection remained relatively constant. In contrast, a DI encoding GFP did not form GFP-positive clusters and the percentage of GFP-positive cells declined by roughly half from 2 to 4 days post-infection. Cluster formation and sustaining the percentage of infected (GFP-positive) cells required the C part of the fusion protein, including the downstream but not the upstream of two arginine clusters (both of which are associated with RNA binding and association with mitochondrial p32 protein) and the E1 part through the transmembrane sequence, but not the C-terminal cytoplasmic tail. Among a collection of mutant DI constructs, cluster formation and sustaining infected cell percentage correlated with maintenance during serial passage with wt RUB. We hypothesize that cluster formation and

  17. 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

  18. 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.

  19. Surface behavior of peptides from E1 GBV-C protein: Interaction with anionic model membranes and importance in HIV-1 FP inhibition.

    PubMed

    Galatola, R; Cruz, A; Gómara, M J; Prat, J; Alsina, M A; Haro, I; Pujol, M

    2015-02-01

    The interaction between a peptide sequence from GB virus C E1 protein (E1P8) and its structural analogs (E1P8-12), (E1P8-13), and (E1P8-21) with anionic lipid membranes (POPG vesicles and POPG, DPPG or DPPC/DPPG (2:1) monolayers) and their association with HIV-1 fusion peptide (HIV-1 FP) inhibition at the membrane level were studied using biophysical methods. All peptides showed surface activity but leakage experiments in vesicles as well as insertion kinetics in monolayers and lipid/peptide miscibility indicated a low level of interaction: neither E1P8 nor its analogs induced the release of vesicular content and the exclusion pressure values (πe) were clearly lower than the biological membrane pressure (24-30 mN m(-1)) and the HIV-1 FP (35 mN m(-1)). Miscibility was elucidated in terms of the additivity rule and excess free energy of mixing (GE). E1P8, E1P8-12 and E1P8-21 (but not E1P8-13) induced expansion of the POPG monolayer. The mixing process is not thermodynamically favored as the positive GE values indicate. To determine how E1 peptides interfere in the action of HIV-1 FP at the membrane level, mixed monolayers of HIV-1 FP/E1 peptides (2:1) and POPG were obtained. E1P8 and its derivative E1P8-21 showed the greatest HIV-1 FP inhibition. The LC-LE phase lipid behavior was morphologically examined via fluorescence microscopy (FM) and atomic force microscopy (AFM). Images revealed that the E1 peptides modify HIV-1 FP-lipid interaction. This fact may be attributed to a peptide/peptide interaction as indicated by AFM results. Finally, hemolysis assay demonstrated that E1 peptides inhibit HIV-1 FP activity.

  20. Conditioned Reinforcement and Response Strength

    PubMed Central

    Shahan, Timothy A

    2010-01-01

    Stimuli associated with primary reinforcers appear themselves to acquire the capacity to strengthen behavior. This paper reviews research on the strengthening effects of conditioned reinforcers within the context of contemporary quantitative choice theories and behavioral momentum theory. Based partially on the finding that variations in parameters of conditioned reinforcement appear not to affect response strength as measured by resistance to change, long-standing assertions that conditioned reinforcers do not strengthen behavior in a reinforcement-like fashion are considered. A signposts or means-to-an-end account is explored and appears to provide a plausible alternative interpretation of the effects of stimuli associated with primary reinforcers. Related suggestions that primary reinforcers also might not have their effects via a strengthening process are explored and found to be worthy of serious consideration. PMID:20885815

  1. 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.

  2. Conditioned reinforcement and response strength.

    PubMed

    Shahan, Timothy A

    2010-03-01

    Stimuli associated with primary reinforcers appear themselves to acquire the capacity to strengthen behavior. This paper reviews research on the strengthening effects of conditioned reinforcers within the context of contemporary quantitative choice theories and behavioral momentum theory. Based partially on the finding that variations in parameters of conditioned reinforcement appear not to affect response strength as measured by resistance to change, long-standing assertions that conditioned reinforcers do not strengthen behavior in a reinforcement-like fashion are considered. A signposts or means-to-an-end account is explored and appears to provide a plausible alternative interpretation of the effects of stimuli associated with primary reinforcers. Related suggestions that primary reinforcers also might not have their effects via a strengthening process are explored and found to be worthy of serious consideration.

  3. An indentation fatigue strength law

    NASA Astrophysics Data System (ADS)

    Xu, Baoxing; Yonezu, Akio; Chen, Xi

    2010-05-01

    Indentation fatigue, where a cyclic load is applied on the sample via an indenter, emerges as an alternative approach for measuring the fatigue properties of materials. We have carried out indentation fatigue tests on a poly(vinyl chloride) (PVC) bulk material, as well as on TiN and NiP films/coatings deposited on SUS304 steel substrates, and demonstrate that a simple power-law relationship can be established between the indentation load amplitude and number of cycles to failure. Such a law is very similar to the conventional fatigue strength law obtained from uniaxial tests. The agreement between the fatigue stress exponents obtained by uniaxial and indentation fatigue tests suggests the potential applicability of the indentation fatigue technique for extracting the fatigue properties of materials.

  4. Conditioned reinforcement and response strength.

    PubMed

    Shahan, Timothy A

    2010-03-01

    Stimuli associated with primary reinforcers appear themselves to acquire the capacity to strengthen behavior. This paper reviews research on the strengthening effects of conditioned reinforcers within the context of contemporary quantitative choice theories and behavioral momentum theory. Based partially on the finding that variations in parameters of conditioned reinforcement appear not to affect response strength as measured by resistance to change, long-standing assertions that conditioned reinforcers do not strengthen behavior in a reinforcement-like fashion are considered. A signposts or means-to-an-end account is explored and appears to provide a plausible alternative interpretation of the effects of stimuli associated with primary reinforcers. Related suggestions that primary reinforcers also might not have their effects via a strengthening process are explored and found to be worthy of serious consideration. PMID:20885815

  5. IGT Stack No. 6 (SDG{ampersand}E-1) Test Plan and Component Specification Document: Topical report, March 1996

    SciTech Connect

    1997-12-31

    The purpose of Stack-6 (SDG{ampersand}E-1) is to scale up and demonstrate the long term performance and endurance characteristics of the IMHEX stack design and the Generation No. 2 cell components (improved pore matching electrodes) in a 20 cell subscale stack test.

  6. IGT Stack No. 6 (SDG[ampersand]E-1) Test Plan and Component Specification Document: Topical report, March 1996

    SciTech Connect

    Not Available

    1997-01-01

    The purpose of Stack-6 (SDG[ampersand]E-1) is to scale up and demonstrate the long term performance and endurance characteristics of the IMHEX stack design and the Generation No. 2 cell components (improved pore matching electrodes) in a 20 cell subscale stack test.

  7. Association of mitochondrial deoxyribonucleic acid mutation with polymorphism in CYP2E1 gene in oral carcinogenesis

    PubMed Central

    Pandey, Rahul; Mehrotra, Divya; Catapano, Carlo; Choubey, Vimal; Sarin, Rajiv; Mahdi, Abbas Ali; Singh, Stuti

    2012-01-01

    Background Oral carcinogenesis is a complex process affected by genetic as well as environmental factors. CYP2E1 gene is involved in metabolism of number of compounds and carcinogens. Its normal functioning is required for homeostasis of free radical. Mitochondrial deoxyribonucleic acid (mtDNA) is 10–100 times more susceptible to damage than nuclear DNA. Mitochondrial DNA large scale deletions are well documented in oral cancer. However, the relationship between CYP2E1 gene polymorphisms and mtDNA damage is still not documented in literature. Materials and Methods Case–control study involving 50 subjects was carried out. Deoxyribonucleic acid extraction was done from study subject tissue samples. Restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR) amplification was done to confirm CYP2E1 gene polymorphisms. The PCR amplification was done for mtDNA 4977 bp deletion. Statistical analysis was carried out using SPSS version 11.5 with χ2 tests. Results c1c1 and DD polymorphisms are prevalent in North Indian population having oral cancer. These polymorphisms are significantly associated with mtDNA 4977 bp deletion. Conclusion Mitochondrial DNA damage induced by wild CYP2E1 forms and imperfect DNA repair in mtDNA may act synergistically to greatly enhance oral cancer risk. PMID:25756024

  8. An amino acid substitution in the pyruvate dehydrogenase E1{alpha} gene, affecting mitochondrial import of the precursor protein

    SciTech Connect

    Takakubo, F.; Thorburn, D.R.; Dahl, H.H.M.

    1995-10-01

    A mutation in the mitochondrial targeting sequence was characterized in a male patient with X chromosome-linked pyruvate dehydrogenase E1{alpha} deficiency. The mutation was a base substitution of G by C at nucleotide 134 in the mitochondrial targeting sequence of the PDHA1 gene, resulting in an arginine-to-proline substitution at codon 10 (R10P). Pyruvate dehydrogenase activity in cultured skin fibroblasts was 28% of the control value, and immunoblot analysis revealed a decreased level of pyruvate dehydrogenase E1{alpha}immunoreactivity. Chimeric constructs in which the normal and mutant pyruvate dehydrogenase E1{alpha} targeting sequences were attached to the mitochondrial matrix protein ornithine transcarbamylase were synthesized in a cell free translation system, and mitochondrial import of normal and mutant proteins was compared in vitro. The results show that ornithine transcarbamylase targeted by the mutant pyruvate dehydrogenase E1{alpha} sequence was translocated into the mitochondrial matrix at a reduced rate, suggesting that defective import is responsible for the reduced pyruvate dehydrogenase level in mitochondria. The mutation was also present in an affected brother and the mildly affected mother. The clinical presentations of this X chromosome-linked disorder in affected family members are discussed. To our knowledge, this is the first report of an amino acid substitution in a mitochondrial targeting sequence resulting in a human genetic disease. 58 refs., 5 figs., 1 tab.

  9. Use of CYP2E1-transfected human liver cell lines in elucidating the actions of ethanol.

    PubMed

    Lakshman, Raj; Cederbaum, Arthur I; Hoek, Jan B; Konishi, Masahiro; Koop, Dennis; Donohu, Terrence M

    2005-09-01

    This article represents the proceedings of a symposium at the 2004 RSA Meeting held in Vancouver, Canada. The chairs were Arthur I. Cederbaum and Raj Lakshman. The presentations were (1) ethanol regulates 2,6-sialyltransferase (2,6-ST) gene expression posttranscriptionally by the interaction of a cytosolic binding protein with 2,6-ST mRNA in CYP2E1- and ADH-transfected HepG2 cells, by Raj Lakshman; (2) nature versus nurture: HepG2-E47 cells as a tool to investigate mechanisms of ethanol-mediated potentiation of cell killing, by Jan B. Hoek; (3) ethanol up-regulates profibrogenic connective tissue growth factor gene expression in HepG2 cells via cytochrome P-450 2E1-mediated ethanol oxidation, by Masahiro Konishi; (4) role of calcium and calcium-activated enzymes in CYP2E1-dependent toxicity, by Arthur I Cederbaum; (5) the use of cell lines to characterize the role of CYP2E1 in the metabolism of farnesol, by Dennis Koop; and (6) studies with HepG2 cells that express the two major ethanol-metabolizing enzymes, by Terrence M. Donohue. PMID:16205373

  10. Mobilization properties of small ColE1-like plasmids carrying kanamycin resistance gene isolated from Salmonella enterica serotypes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Previously we isolated and characterized various groups of small kanamycin resistance (KanR) ColE1-like plasmids from different serotypes of Salmonella enterica isolates. These plasmids all carried the aph(3)-I gene encoding the aminoglycoside phosphotransferase responsible for the kanam...

  11. An improved HAdV-41 E1B55K-expressing 293 cell line for packaging fastidious adenovirus.

    PubMed

    Zou, Xiao-Hui; Xiao, Xia; Chen, Duo-Ling; Li, Ze-Liang; Song, Jing-Dong; Wang, Min; Qu, Jian-Guo; Lu, Zhuo-Zhuang; Hung, Tao

    2011-08-01

    Human adenovirus type 41 (HAdV-41) is difficult to cultivate under laboratory conditions. Tripartite leader sequence (TPL) of HAdV-41 was cloned, inserted into eukaryotic expression plasmid, and used to establish a HAdV-41 E1B55K-transduced cell line (293TE7). HAdV-41 E1B55K was expressed more abundantly in 293TE7 than in 293E12, an HAdV-41 E1B55K-expressing cell line developed previously. After being infected with E1-deleted HAdV-41 vector (HAdV-41-GFP), 293TE7 synthesized more viral genomic DNA and structural proteins, which led ultimately to a significant increase of the yield of progeny viruses. Typically, 293TE7 produced progeny viruses 3-15 times more than 293E12 did, depending on the amount of seed viruses and culture time. These data demonstrated that 293TE7 was an effective packaging cell line, and implied its application for wild-type HAdV-41 isolation, HAdV-41 virological study and recombinant HAdV-41 construction. PMID:21601594

  12. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 26 Internal Revenue 18 2012-04-01 2012-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  13. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 26 Internal Revenue 18 2014-04-01 2014-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  14. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 26 Internal Revenue 18 2013-04-01 2013-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  15. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 26 Internal Revenue 18 2011-04-01 2011-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  16. 26 CFR 301.6503(e)-1 - Suspension of running of period of limitation; certain powers of appointment.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 26 Internal Revenue 18 2010-04-01 2010-04-01 false Suspension of running of period of limitation... Limitations Limitations on Assessment and Collection § 301.6503(e)-1 Suspension of running of period of... complied with, the running of the period of limitation for assessment or collection of any estate...

  17. Effects of herbal products and their constituents on human cytochrome P450(2E1) activity.

    PubMed

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

    2007-12-01

    Ethanolic extracts from fresh Echinacea purpurea and Spilanthes acmella and dried Hydrastis canadensis were examined with regard to their ability to inhibit cytochrome P450(2E1) 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 P450(2E1), 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 K(I) values ranging from 2.8 microM for hydrastine to 18 microM for berberine. The alkylamides present in E. purpurea and S. acmella also showed significant inhibition at concentrations as low as 25 microM, 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 P450(2E1). PMID:17658211

  18. 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

    ... TAXES (CONTINUED) Items Specifically Included in Gross Income § 1.72(e)-1T Treatment of distributions... employee contributions and thus were not includible in gross income. After distributions equaled the... and, second, as allocable to nondeductible employee contributions. Distributions allocable to...

  19. Ghrelin inhibits the apoptosis of MC3T3-E1 cells through ERK and AKT signaling pathway

    SciTech Connect

    Liang, Qiu-Hua; Liu, Yuan; Wu, Shan-Shan; Cui, Rong-Rong; Yuan, Ling-Qing Liao, Er-Yuan

    2013-11-01

    Ghrelin is a 28-amino-acid peptide that acts as a natural endogenous ligand of the growth hormone secretagogue receptor (GHSR) and strongly stimulates the release of growth hormone from the hypothalamus–pituitary axis. Previous studies have identified the important physiological effects of ghrelin on bone metabolism, such as regulating proliferation and differentiation of osteoblasts, independent of GH/IGF-1 axis. However, research on effects and mechanisms of ghrelin on osteoblast apoptosis is still rare. In this study, we identified expression of GHSR in MC3T3-E1 cells and determined the effects of ghrelin on the apoptosis of osteoblastic MC3T3-E1 cells and the mechanism involved. Our data demonstrated that ghrelin inhibited the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, as determined by terminal deoxynucleotidyl transferase-mediated deoxyribonucleotide triphosphate nick end-labeling (TUNEL) and ELISA assays. Moreover, ghrelin upregulated Bcl-2 expression and downregulated Bax expression in a dose-dependent manner. Our study also showed decreased activated caspase-3 activity under the treatment of ghrelin. Further study suggested that ghrelin stimulated the phosphorylation of ERK and AKT. Pretreatment of cells with the ERK inhibitor PD98059, PI3K inhibitor LY294002, and GHSR-siRNA blocked the ghrelin-induced activation of ERK and AKT, respectively; however, ghrelin did not stimulate the phosphorylation of p38 or JNK. PD90859, LY294002 and GHSR-siRNA attenuated the anti-apoptosis effect of ghrelin in MC3T3-E1 cells. In conclusion, ghrelin inhibits the apoptosis of osteoblastic MC3T3-E1 cells induced by serum deprivation, which may be mediated by activating the GHSR/ERK and GHSR/PI3K/AKT signaling pathways. - Highlights: • We explored the effects of ghrelin on serum deprivation-induced MC3T3-E1 cells apoptosis. • Both ELISA and TUNEL were used to detect the apoptosis. • The receptor of ghrelin, GHSR, was expressed in MC3T3-E1

  20. 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…

  1. Crystal Structure of UBA2ufd-Ubc9: Insights into E1-E2 Interactions in Sumo Pathways

    PubMed Central

    Hunt, Harold W.; Seyedin, Steven N.; Miller, David W.; Miller, Darcie J.; Huang, Danny T.; Schulman, Brenda A.

    2010-01-01

    Canonical ubiquitin-like proteins (UBLs) such as ubiquitin, Sumo, NEDD8, and ISG15 are ligated to targets by E1-E2-E3 multienzyme cascades. The Sumo cascade, conserved among all eukaryotes, regulates numerous biological processes including protein localization, transcription, DNA replication, and mitosis. Sumo conjugation is initiated by the heterodimeric Aos1-Uba2 E1 enzyme (in humans called Sae1-Uba2), which activates Sumo's C-terminus, binds the dedicated E2 enzyme Ubc9, and promotes Sumo C-terminal transfer between the Uba2 and Ubc9 catalytic cysteines. To gain insights into details of E1-E2 interactions in the Sumo pathway, we determined crystal structures of the C-terminal ubiquitin fold domain (ufd) from yeast Uba2 (Uba2ufd), alone and in complex with Ubc9. The overall structures of both yeast Uba2ufd and Ubc9 superimpose well on their individual human counterparts, suggesting conservation of fundamental features of Sumo conjugation. Docking the Uba2ufd-Ubc9 and prior full-length human Uba2 structures allows generation of models for steps in Sumo transfer from Uba2 to Ubc9, and supports the notion that Uba2 undergoes remarkable conformational changes during the reaction. Comparisons to previous structures from the NEDD8 cascade demonstrate that UBL cascades generally utilize some parallel E1-E2 interaction surfaces. In addition, the structure of the Uba2ufd-Ubc9 complex reveals interactions unique to Sumo E1 and E2. Comparison with a previous Ubc9-E3 complex structure demonstrates overlap between Uba2 and E3 binding sites on Ubc9, indicating that loading with Sumo and E3-catalyzed transfer to substrates are strictly separate steps. The results suggest mechanisms establishing specificity and order in Sumo conjugation cascades. PMID:21209884

  2. Gene order for rubella virus structural proteins is NH/sub 2/-C-E2-E1-COOH

    SciTech Connect

    Oker-Blom, C.

    1984-08-01

    The order of translation in vivo of the genes coding for rubella virus structural proteins was studied in infected B-Vero cells. The proteins were sequentially pulse-chase labeled with (/sup 35/S)methionine after synchronization of translation initiation with hypertonic salt treatment. A sequential labeling procedure (window-labeling) to specifically label defined segments of the structural proteins was also used. The labeled proteins were identified by sodium dodecyl sulfate-gel electrophoresis after immunoprecipitation with specific antisera directed against the two virion glycoproteins (E1 and E2a/E2b) and the nucleocapsid (C) protein. The order of translation was found to be NH/sub 2/-C-E2-E1-COOH. We have previously shown that the structural proteins are synthesized in vitro from a cytoplasmic 24S subgenomic mRNA as a 110,000-dalton (p110) precursor. Here, it is shown that p110 is precipitated with anti-C, anti-E2, and anti-E1 sera, indicating that p110 is the precursor of all three structural proteins. Two major in vitro translation products (M/sub r/s, 66,000 and 62,000) that could represent preterminated polypeptide chains or proteolytic cleavage products were precipitated with anti-C and anti-Es sera, but not with anti-E1 serum, indicating, in conformity with the in vivo results, that the genes for the C and E2 proteins are adjacent to each other. Using these specific antisera, we have also confirmed the identity of the unglycosylated forms of E1 (M/sub r/, 53,000) and E2 (M/sub r/ 30,000) immunoprecipitated from tunicamycin-treated infected cells. 18 references, 6 figures.

  3. Structural Basis for Cooperative Binding of Azoles to CYP2E1 as Interpreted through Guided Molecular Dynamics Simulations

    PubMed Central

    Levy, Joseph W.; Hartman, Jessica H.; Perry, Martin D.; Miller, Grover P.

    2015-01-01

    CYP2E1 metabolizes a wide array of small, hydrophobic molecules, resulting in their detoxification or activation into carcinogens through Michaelis-Menten as well as cooperative mechanisms. Nevertheless, the molecular determinants for CYP2E1 specificity and metabolic efficiency toward these compounds are still unknown. Herein, we employed computational docking studies coupled to Molecular Dynamics simulations to provide a critical perspective for understanding a structural basis for cooperativity observed for an array of azoles from our previous binding and catalytic studies (Hartman, JH et al (2014) Biochem Pharmacol 87, 523-33). The resulting 28 CYP2E1 complexes in this study revealed a common passageway for azoles that included a hydrophobic steric barrier causing a pause in movement toward the active site. The entrance to the active site acted like a second sieve to restrict access to the inner chamber. Collectively, these interactions impacted the final orientation of azoles reaching the active site and hence could explain differences in their biochemical properties observed in our previous studies, such as the consequences of methylation at position 5 of the azole ring. The association of a second azole demonstrated significant differences in interactions stabilizing the bound complex than observed for the first binding event. Intermolecular interactions occurred between the two azoles as well as CYP2E1 residue side chains and backbone and involved both hydrophobic contacts and hydrogen bonds. The relative importance of these interactions depended on the structure of the respective azoles indicating the absence of specific defining criteria for binding unlike the well-characterized dominant role of hydrophobicity in active site binding. Consequently, the structure activity relationships described here and elsewhere are necessary to more accurately identify factors impacting the observation and significance of cooperativity in CYP2E1 binding and catalysis

  4. Atmospheric volatilization and distribution of (Z)- and (E)-1,3-dichloropropene in field beds with and without plastic covers.

    PubMed

    Thomas, John E; Allen, L Hartwell; McCormack, Leslie A; Vu, Joseph C; Dickson, Donald W; Ou, Li-Tse

    2004-01-01

    The fumigant 1,3-dichloropropene (1,3-D) is considered to be a potential replacement for methyl bromide when methyl bromide is phased out in 2005. This study on surface emissions and subsurface diffusion of 1,3-D in a Florida sandy soil was conducted in field beds with or without plastic covers. After injection of the commercial fumigant Telone II by conventional chisels to field beds at 30cm depth which were covered with polyethylene film (PE), virtually impermeable film, or no cover (bare), (Z)- and (E)-1,3-D rapidly diffused upward. Twenty hours after injection, majority of (Z)- and (E)-1,3-D had moved upward from 30 cm depth to the layer of 5-20 cm depth. Downward movement of the two isomers in the beds with or without a plastic cover was not significant. (Z)-1,3-D diffused more rapidly than (E)-1,3-D. Virtually impermeable films (VIF) had a good capacity to retain (Z)- and (E)-1,3-D in soil pore air space. Vapor concentrations of the two isomers in the shallow subsurface of the field bed covered with VIF were greater than that in the two beds covered with polyethylene film (PE) or no cover (bare). In addition, VIF cover provided more uniform distribution of (Z)- and (E)-1,3-D in shallow subsurface than PE cover or no cover. Virtually impermeable film also had a better capability to retard surface emissions of the two isomers from soil in field beds than PE cover or no cover.

  5. Bovine Collagen Peptides Compounds Promote the Proliferation and Differentiation of MC3T3-E1 Pre-Osteoblasts

    PubMed Central

    Liu, JunLi; Zhang, Bing; Song, ShuJun; Ma, Ming; Si, ShaoYan; Wang, YiHu; Xu, BingXin; Feng, Kai; Wu, JiGong; Guo, YanChuan

    2014-01-01

    Objective Collagen peptides (CP) compounds, as bone health supplements, are known to play a role in the treatment of osteoporosis. However, the molecular mechanisms of this process remain unclear. This study aimed to investigate the effects of bovine CP compounds on the proliferation and differentiation of MC3T3-E1 cells. Methods Mouse pre-osteoblast cell line MC3T3-E1 subclone 4 cells were treated with bovine CP compounds. Cell proliferation was analyzed by MTT assays and the cell cycle was evaluated by flow cytometry scanning. Furthermore, MC3T3-E1 cell differentiation was analyzed at the RNA level by real-time PCR and at the protein level by western blot analysis for runt-related transcription factor 2 (Runx2), a colorimetric p-nitrophenyl phosphate assay for alkaline phosphatase (ALP), and ELISA for osteocalcin (OC). Finally, alizarin red staining for mineralization was measured using Image Software Pro Plus 6.0. Results Cell proliferation was very efficient after treatment with different concentrations of bovine CP compounds, and the best concentration was 3 mg/mL. Bovine CP compounds significantly increased the percentage of MC3T3-E1 cells in G2/S phase. Runx2 expression, ALP activity, and OC production were significantly increased after treatment with bovine CP compounds for 7 or 14 days. Quantitative analyses with alizarin red staining showed significantly increased mineralization of MC3T3-E1 cells after treatment with bovine CP compounds for 14 or 21 days. Conclusions Bovine CP compounds increased osteoblast proliferation, and played positive roles in osteoblast differentiation and mineralized bone matrix formation. Taking all the experiments together, our study indicates a molecular mechanism for the potential treatment of osteoarthritis and osteoporosis. PMID:24926875

  6. Crystal Structure of UBA2[superscript ufd]-Ubc9: Insights into E1-E2 Interactions in Sumo Pathways

    SciTech Connect

    Wang, Jing; Taherbhoy, Asad M.; Hunt, Harold W.; Seyedin, Steven N.; Miller, David W.; Miller, Darcie J.; Huang, Danny T.; Schulman, Brenda A.

    2012-04-30

    Canonical ubiquitin-like proteins (UBLs) such as ubiquitin, Sumo, NEDD8, and ISG15 are ligated to targets by E1-E2-E3 multienzyme cascades. The Sumo cascade, conserved among all eukaryotes, regulates numerous biological processes including protein localization, transcription, DNA replication, and mitosis. Sumo conjugation is initiated by the heterodimeric Aos1-Uba2 E1 enzyme (in humans called Sae1-Uba2), which activates Sumo's C-terminus, binds the dedicated E2 enzyme Ubc9, and promotes Sumo C-terminal transfer between the Uba2 and Ubc9 catalytic cysteines. To gain insights into details of E1-E2 interactions in the Sumo pathway, we determined crystal structures of the C-terminal ubiquitin fold domain (ufd) from yeast Uba2 (Uba2{sup ufd}), alone and in complex with Ubc9. The overall structures of both yeast Uba2{sup ufd} and Ubc9 superimpose well on their individual human counterparts, suggesting conservation of fundamental features of Sumo conjugation. Docking the Uba2{sup ufd}-Ubc9 and prior full-length human Uba2 structures allows generation of models for steps in Sumo transfer from Uba2 to Ubc9, and supports the notion that Uba2 undergoes remarkable conformational changes during the reaction. Comparisons to previous structures from the NEDD8 cascade demonstrate that UBL cascades generally utilize some parallel E1-E2 interaction surfaces. In addition, the structure of the Uba2{sup ufd}-Ubc9 complex reveals interactions unique to Sumo E1 and E2. Comparison with a previous Ubc9-E3 complex structure demonstrates overlap between Uba2 and E3 binding sites on Ubc9, indicating that loading with Sumo and E3-catalyzed transfer to substrates are strictly separate steps. The results suggest mechanisms establishing specificity and order in Sumo conjugation cascades.

  7. Resolvin E1 analog RX-10045 0.1% reduces corneal stromal haze in rabbits when applied topically after PRK

    PubMed Central

    Torricelli, Andre A. M.; Santhanam, Abirami; Agrawal, Vandana

    2014-01-01

    Purpose To perform a masked study to determine whether resolvin E1 (RvE1), a lipid-derived immunomodulator, could regulate the development of corneal haze and opacity-related myofibroblasts after opacity-generating high correction photorefractive keratectomy (PRK) in rabbits. Methods Three groups of eight rabbits each were included in the study. Nine diopter (D) PRK for myopia was performed in each test cornea, and the eyes were treated with 30 µl of topical solution every 4 h (six times a day) for 5 days starting immediately after PRK. Group 1 was treated with 0.1% RX-10045, a prodrug of an RvE1 analog; group 2 was treated with 0.01% RX-10045; and group 3 was treated with vehicle control solution. At 1 month after PRK, haze was graded at the slit-lamp by a masked observer. Immunohistochemistry for α-smooth muscle actin (SMA) was performed on the central cornea of each test eye to determine the anterior stromal myofibroblast density. Results Corneal opacity was significantly lower in the 0.1% RX-10045 group, but not the 0.01% RX-10045 group, compared to the vehicle control group (p=0.029), at 1 month after −9.0D PRK. At 1 month after −9.0D PRK, SMA+ myofibroblast densities in the anterior stroma were not statistically significantly different among the three groups, although a trend toward lower myofibroblast generation was noted in the 0.1% RX-10045 group. Conclusions Topical 0.1% RX-10045, a prodrug of an RvE1 analog, reduces corneal opacity after haze-generating PRK in rabbits. Further studies are needed to determine the precise points at which RvE1 decreases corneal opacity after injury. PMID:25558174

  8. Low-Level Expression of the E1B 20-Kilodalton Protein by Adenovirus 14p1 Enhances Viral Immunopathogenesis.

    PubMed

    Radke, Jay R; Yong, Sherri L; Cook, James L

    2016-01-01

    Adenovirus 14p1 (Ad14p1) is an emergent variant of Ad serotype 14 (Ad14) that has caused increased severity of respiratory illnesses during globally distributed outbreaks, including cases of acute respiratory distress syndrome and death. We found that human cell infection with Ad14p1 results in markedly decreased expression of the E1B 20-kilodalton (20K) protein compared to that with infection with wild-type (wt) Ad14. This reduced Ad14p1 E1B 20K expression caused a loss-of-function phenotype of Ad-infected cell corpses that, in contrast to cells infected with wt Ad14, either failed to repress or increased NF-κB-dependent, proinflammatory cytokine responses of responder human alveolar macrophages. A small-animal model of Ad14-induced lung infection was used to test the translational relevance of these in vitro observations. Intratracheal infection of Syrian hamsters with Ad14p1 caused a marked, patchy bronchopneumonia, whereas hamster infection with wt Ad14 caused minimal peribronchial inflammation. These results suggest that this difference in E1B 20K gene expression during Ad14p1 infection and its modulating effect on the interactions between Ad14-infected cells and the host innate immune response could explain the increased immunopathogenic potential and associated increase in clinical illness in some people infected with the Ad14p1 outbreak strain.IMPORTANCE We previously reported that Ad-infected human cells exhibit E1B 19K-dependent repression of virally induced, NF-κB-dependent macrophage cytokine responses (J. R. Radke, F. Grigera, D. S. Ucker, and J. L. Cook, J Virol 88:2658-2669, 2014, http://dx.doi.org/10.1128/JVI.02372-13). The more virulent, emergent strain of Ad14, Ad14p1, causes increased cytopathology in vitro, which suggested a possible E1B 20K defect. Whether there is a linkage between these observations was unknown. We show that there is markedly reduced expression of E1B 20K in Ad14p1-infected human cells and that this causes an increased

  9. Low-Level Expression of the E1B 20-Kilodalton Protein by Adenovirus 14p1 Enhances Viral Immunopathogenesis

    PubMed Central

    Yong, Sherri L.; Cook, James L.

    2015-01-01

    ABSTRACT Adenovirus 14p1 (Ad14p1) is an emergent variant of Ad serotype 14 (Ad14) that has caused increased severity of respiratory illnesses during globally distributed outbreaks, including cases of acute respiratory distress syndrome and death. We found that human cell infection with Ad14p1 results in markedly decreased expression of the E1B 20-kilodalton (20K) protein compared to that with infection with wild-type (wt) Ad14. This reduced Ad14p1 E1B 20K expression caused a loss-of-function phenotype of Ad-infected cell corpses that, in contrast to cells infected with wt Ad14, either failed to repress or increased NF-κB-dependent, proinflammatory cytokine responses of responder human alveolar macrophages. A small-animal model of Ad14-induced lung infection was used to test the translational relevance of these in vitro observations. Intratracheal infection of Syrian hamsters with Ad14p1 caused a marked, patchy bronchopneumonia, whereas hamster infection with wt Ad14 caused minimal peribronchial inflammation. These results suggest that this difference in E1B 20K gene expression during Ad14p1 infection and its modulating effect on the interactions between Ad14-infected cells and the host innate immune response could explain the increased immunopathogenic potential and associated increase in clinical illness in some people infected with the Ad14p1 outbreak strain. IMPORTANCE We previously reported that Ad-infected human cells exhibit E1B 19K-dependent repression of virally induced, NF-κB-dependent macrophage cytokine responses (J. R. Radke, F. Grigera, D. S. Ucker, and J. L. Cook, J Virol 88:2658–2669, 2014, http://dx.doi.org/10.1128/JVI.02372-13). The more virulent, emergent strain of Ad14, Ad14p1, causes increased cytopathology in vitro, which suggested a possible E1B 20K defect. Whether there is a linkage between these observations was unknown. We show that there is markedly reduced expression of E1B 20K in Ad14p1-infected human cells and that this causes an

  10. Arg-Gly-Asp (RGD)-Modified E1A/E1B Double Mutant Adenovirus Enhances Antitumor Activity in Prostate Cancer Cells In Vitro and in Mice.

    PubMed

    Shen, Yue-Hong; Yang, Fei; Wang, Hua; Cai, Zhi-Jian; Xu, Yi-Peng; Zhao, An; Su, Ying; Zhang, Gu; Zhu, Shao-Xing

    2016-01-01

    CAR is a transmembrane protein that is expressed in various epithelial and endothelial cells. CAR mediates adenoviral infection, as well as adenovirus-mediated oncolysis of AxdAdB-3, an E1A/E1B double-restricted oncolytic adenovirus, in prostate cancer cells. This study further assessed the therapeutic efficacy of AxdAdB-3 with Arg-Gly-Asp (RGD)-fiber modification (AxdAdB3-F/RGD), which enables integrin-dependent infection, in prostate cancer. Susceptibility of prostate cancer cells LNCaP, PC3, and DU145 to adenovirus infection was associated with CAR expression. All of the prostate cancer cell lines expressed integrin αvβ3 and αvβ5. AxdAdB-3 was more cytopathic in CAR-positive prostate cancer cells than in CAR-negative cells, whereas AxdAdB3-F/RGD caused potent oncolysis in both CAR-positive and CAR-negative prostate cancer cells. In contrast, AxdAdB3-F/RGD was not cytopathic against normal prostate epithelial cells, RWPE-1. Intratumoral injection of AxdAdB3-F/RGD into CAR-negative prostate cancer cell xenografts in nude mice inhibited tumor growth. The current study demonstrates that E1A/E1B double-restricted oncolytic adenovirus with an RGD-fiber modification enhances infection efficiency and anti-tumor activity in CAR-deficient prostate cancer cells, while sparing normal cells. Future studies will evaluate the therapeutic potential of AxdAdB3-F/RGD in prostate cancer. PMID:26799485

  11. Inhibition of human cytochrome P450 2E1 and 2A6 by aldehydes: structure and activity relationships.

    PubMed

    Kandagatla, Suneel K; Mack, Todd; Simpson, Sean; Sollenberger, Jill; Helton, Eric; Raner, Gregory M

    2014-08-01

    The purpose of this study was to probe active site structure and dynamics of human cytochrome P4502E1 and P4502A6 using a series of related short chain fatty aldehydes. Binding efficiency of the aldehydes was monitored via their ability to inhibit the binding and activation of the probe substrates p-nitrophenol (2E1) and coumarin (2A6). Oxidation of the aldehydes was observed in reactions with individually expressed 2E1, but not 2A6, suggesting alternate binding modes. For saturated aldehydes the optimum chain length for inhibition of 2E1 was 9 carbons (KI=7.8 ± 0.3 μM), whereas for 2A6 heptanal was most potent (KI=15.8 ± 1.1 μM). A double bond in the 2-position of the aldehyde significantly decreased the observed KI relative to the corresponding saturated compound in most cases. A clear difference in the effect of the double bond was observed between the two isoforms. With 2E1, the double bond appeared to remove steric constraints on aldehyde binding with KI values for the 5-12 carbon compounds ranging between 2.6 ± 0.1 μM and 12.8 ± 0.5 μM, whereas steric effects remained the dominant factor in the binding of the unsaturated aldehydes to 2A6 (observed KI values between 7.0 ± 0.5 μM and >1000 μM). The aldehyde function was essential for effective inhibition, as the corresponding carboxylic acids had very little effect on enzyme activity over the same range of concentrations, and branching at the 3-position of the aldehydes increased the corresponding KI value in all cases examined. The results suggest that a conjugated π-system may be a key structural determinant in the binding of these compounds to both enzymes, and may also be an important feature for the expansion of the active site volume in 2E1.

  12. Inhibition of human Cytochrome P450 2E1 and 2A6 by aldehydes: Structure and activity relationships

    PubMed Central

    Kandagatla, Suneel K.; Mack, Todd; Simpson, Sean; Sollenberger, Jill; Helton, Eric; Raner, Gregory M.

    2014-01-01

    The purpose of this study was to probe active site structure and dynamics of human cytochrome P4502E1 and P4502A6 using a series of related short chain fatty aldehydes. Binding efficiency of the aldehydes was monitored via their ability to inhibit the binding and activation of the probe substrates p-nitrophenol (2E1) and coumarin (2A6). Oxidation of the aldehydes was observed in reactions with individually expressed 2E1, but not 2A6, suggesting alternate binding modes. For saturated aldehydes the optimum chain length for inhibition of 2E1 was 9 carbons (KI=7.8 ±0.3 μM), whereas for 2A6 heptanal was most potent (KI=15.8 ±1.1 μM). A double bond in the 2-position of the aldehyde significantly decreased the observed KI relative to the corresponding saturated compound in most cases. A clear difference in the effect of the double bond was observed between the two isoforms. With 2E1, the double bond appeared to remove steric constraints on aldehyde binding with KI values for the 5–12 carbon compounds ranging between 2.6 ± 0.1 μM and 12.8± 0.5 μM, whereas steric effects remained the dominant factor in the binding of the unsaturated aldehydes to 2A6 (observed KI values between 7.0± 0.5 μM and >1000 μM). The aldehyde function was essential for effective inhibition, as the corresponding carboxylic acids had very little effect on enzyme activity over the same range of concentrations, and branching at the 3-position of the aldehydes increased the corresponding KI value in all cases examined. The results suggest that a conjugated π-system may be a key structural determinant in the binding of these compounds to both enzymes, and may also be an important feature for the expansion of the active site volume in 2E1. PMID:24924949

  13. Control of adenovirus E1B mRNA synthesis by a shift in the activities of RNA splice sites.

    PubMed Central

    Montell, C; Fisher, E F; Caruthers, M H; Berk, A J

    1984-01-01

    The primary transcript from adenovirus 2 early region 1B (E1B) is processed by differential RNA splicing into two overlapping mRNAs, 13S and 22S. The 22S mRNA is the major E1B mRNA during the early phase of infection, whereas the 13S mRNA predominates during the late phase. In previous work, it has been shown that this shift in proportions of the E1B mRNAs is influenced by increased cytoplasmic stability of the 13S mRNA at late times in infection. Two observations presented here demonstrate that the increase in proportion of the 13S mRNA at late times is also regulated by a change in the specificity of RNA splicing. First, the relative concentrations of the 13S to 22S nuclear RNAs were not constant throughout infection but increased at late times. Secondly, studies with the mutant, adenovirus 2 pm2250 , provided evidence that there was an increased propensity to utilize a 5' splice in the region of the 13S 5' splice site at late times in infection. Adenovirus 2 pm2250 has a G----C transversion in the first base of E1B 13S mRNA intron preventing splicing of the 13S mRNA but not of the 22S mRNA. During the early phase of a pm2250 infection, the E1B primary transcripts were processed into the 22S mRNA only. However, during the late phase, when the 13S mRNA normally predominates, E1B primary transcripts were also processed by RNA splicing at two formerly unused or cryptic 5' splice sites. Both cryptic splice sites were located much closer to the disrupted 13S 5' splice site than to the 22S 5' splice site. Thus, the temporal increase in proportion of the 13S mRNA to the 22S mRNA is regulated by two processes, an increase in cytoplasmic stability of the 13S mRNA and an increased propensity to utilize the 13S 5' splice site during the late phase of infection. Adenovirus 2 pm2250 was not defective for productive infection of HeLa cells or for transformation of rat cells. Images PMID:6727875

  14. Converging finite-strength shocks

    NASA Astrophysics Data System (ADS)

    Axford, R. A.; Holm, D. D.

    1981-01-01

    The converging shock problem was first solved by Guderley and later by Landau and Stanyukovich for infinitely strong shocks in an ideal gas with spherical and cylindrical symmetry. This problem is solved herein for finite-strength shocks and a non-ideal-gas equation of state with an adiabatic bulk modulus of the type Bs= {- v∂ p}/{∂ v| s} = ( p +B) f( v) , where B is a constant with the dimensions of pressure, and f(v) is an arbitrary function of the specific volume. Self-similar profiles of the particle velocity and thermodynamic variables are studied explicitly for two cases with constant specific heat at constant volume; the Tait-Kirkwood-Murnaghan equation, f(v) = constant, and the Walsh equation, f(v) = v/A, where A = constant. The first case reduces to the ideal gas when B = 0. In both cases the flow behind the shock front exhibits an unbalanced buoyant force instability at a critical Mach number which depends upon equation-of-state parameters.

  15. Dynamic Strength Ceramic Nanocomposites Under Pulse Loading

    NASA Astrophysics Data System (ADS)

    Skripnyak, Evgeniya G.; Skripnyak, Vladimir V.; Vaganova, Irina K.; Skripnyak, Vladimir A.

    2015-06-01

    Multi-scale computer simulation approach has been applied to research of strength of nanocomposites under dynamic loading. The influence of mesoscopic substructures on the dynamic strength of ceramic and hybrid nanocomposites, which can be formed using additive manufacturing were numerically investigated. At weak shock wave loadings the shear strength and the spall strength of ceramic and hybrid nanocomposites depends not only phase concentration and porosity, but size parameters of skeleton substructures. The influence of skeleton parameter on the shear strength and the spall strength of ceramic nanocomposites with the same concentration of phases decreases with increasing amplitude of the shock pulse of microsecond duration above the double amplitude of the Hugoniot elastic limit of nanocomposites. This research carried out in 2014 -2015 was supported by grant from The Tomsk State University Academic D.I. Mendeleev Fund Program and also Ministry of Sciences and Education of Russian Federation (State task 2014/223, project 1943, Agreement 14.132.

  16. Fracture toughness and strength of 96% alumina

    SciTech Connect

    Price, D.B.; Chinn, R.E.; McNerney, K.R.; Brog, T.K.; Kim, C.Y.; Krutyholowa, M.W.; Chen, N.W.; Haun, M.J.

    1997-05-01

    There exists a need to understand the controlling factors that simultaneously impact strength and toughness in 96% alumina. The enhancement of both strength and toughness enables designers to extend the use limits and reliability for structural ceramics. This article presents mechanical property results from a group study examining the use of different alkaline-earth aluminosilicate intergranular compositions containing magnesium, calcium and strontium oxides (RO) in 96% alumina. Principal results address trends in indentation strength toughness and modulus of rupture. Trends in the data are presented relative to existing theories of thermal expansion mismatch toughening, grain-bridging crack-wake effect and crack deflection mechanisms. Strength is addressed in terms of strength after indentation, crack growth of indentation flaws and Weibull characterization for the strength distribution.

  17. Dissection of the C-terminal region of E1A redefines the roles of CtBP and other cellular targets in oncogenic transformation.

    PubMed

    Cohen, M J; Yousef, A F; Massimi, P; Fonseca, G J; Todorovic, B; Pelka, P; Turnell, A S; Banks, L; Mymryk, J S

    2013-09-01

    Human adenovirus E1A makes extensive connections with the cellular protein interaction network. By doing so, E1A can manipulate many cellular programs, including cell cycle progression. Through these reprogramming events, E1A functions as a growth-promoting oncogene and has been used extensively to investigate mechanisms contributing to oncogenesis. Nevertheless, it remains unclear how the C-terminal region of E1A contributes to oncogenic transformation. Although this region is required for transformation in cooperation with E1B, it paradoxically suppresses transformation in cooperation with activated Ras. Previous analysis has suggested that the interaction of E1A with CtBP plays a pivotal role in both activities. However, some C-terminal mutants of E1A retain CtBP binding and yet exhibit defects in transformation, suggesting that other targets of this region are also necessary. To explore the roles of these additional factors, we performed an extensive mutational analysis of the C terminus of E1A. We identified key residues that are specifically required for binding all known targets of the C terminus of E1A. We further tested each mutant for the ability to both localize to the nucleus and transform primary rat cells in cooperation with E1B-55K or Ras. Interaction of E1A with importin α3/Qip1, dual-specificity tyrosine-regulated kinase 1A (DYRK1A), HAN11, and CtBP influenced transformation with E1B-55K. Interestingly, the interaction of E1A with DYRK1A and HAN11 appeared to play a role in suppression of transformation by activated Ras whereas interaction with CtBP was not necessary. This unexpected result suggests a need for revision of current models and provides new insight into transformation by the C terminus of E1A. PMID:23864635

  18. Fracture strength of silicon solar cells

    NASA Technical Reports Server (NTRS)

    Chen, C. P.

    1979-01-01

    A test program was developed to determine the nature and source of the flaw controlling the fracture of silicon solar cells and to provide information regarding the mechanical strength of cells. Significant changes in fracture strengths were found in seven selected in-process wafer-to-cell products from a manufacturer's production line. The fracture strength data were statistically analyzed and interpreted in light of the exterior flaw distribution of the samples.

  19. Transcription of interferon-stimulated genes is induced by adenovirus particles but is suppressed by E1A gene products.

    PubMed Central

    Reich, N; Pine, R; Levy, D; Darnell, J E

    1988-01-01

    Interferon treatment of cell cultures results in the rapid transcriptional induction of a specific set of genes. In this paper we explore the effect of cellular infection by several adenoviruses, both wild type and mutant, on the expression of these genes. Infection with adenovirus induces the transcription of the interferon-stimulated genes in the absence of any protein synthesis. In fact, the inhibition of protein synthesis during a wild-type infection produces enhanced stimulation of transcription of these genes. Experiments with viral mutants indicate the ability to specifically suppress this transcription maps to the E1A gene. In addition, the E1A gene products are capable of suppressing the specific transcriptional induction of interferon-stimulated promoters during cotransfection experiments and therefore presumably during viral infection. The dual effect of adenovirus on the expression of interferon-stimulated genes may represent an example of action and evolutionary reaction between virus and host. Images PMID:2446013

  20. Mechanical strength and stability of lithium aluminate

    NASA Astrophysics Data System (ADS)

    Brimhall, J. L.

    1992-06-01

    Pacific Northwest Laboratory (PNL) investigated the strength and resistance to thermal shock of lithium aluminate annular pellets. The room temperature, axial compressive fracture strength of pellets made at Westinghouse Advanced Energy Systems (WAES) varied from 80 to 133 ksi. The strength at 430 C (806 F) was to 30 to 40 percent lower. The strength at 900 C (1652 F) showed a wide variation with one measurement near 90 ksi. These strength values are consistent with other data and predictions made in the literature when the grain size and porosity of the microstructure are taken into account. In diametral compression tests, the fracture strengths were much lower due to the existence of tensile stresses in some pellet regions from this type of loading. However, the fracture stresses were still generally higher than those reported in the literature; this fracture resistance probably reflects the better quality of the pellets tested in this study. Measurements on pellets made at PNL indicated lower strengths compared to the WAES material. This strength difference could be accounted for by different processing technologies: material made at PNL was cold-pressed and sintered with high porosity whereas the WAES material was isostatically hot-pressed with high density. Thermal shocking of the material by ramping to 900 C in two minutes did not have an observable effect on the microstructure or the strength of any of the pellets.