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Sample records for bright ns lmxbs

  1. INTEGRAL long-term monitoring results on persistently bright NS LMXBs

    NASA Astrophysics Data System (ADS)

    Savolainen, P.; Hannikainen, D. C.; Paizis, A.; Farinelli, R.; Kuulkers, E.; Vilhu, O.

    2010-07-01

    We present long-term spectral and timing results from an INTEGRAL monitoring program of persistently bright neutron star Low-Mass X-ray Binaries, i.e. the three bright Atoll sources GX 3+1, GX 9+1 and GX 9+9, and the Z sources GX 5-1, GX 17+2, GX 340+0 and GX 349+2. From the available observing periods between 2003 and 2009, each lasting ~2 months, we have selected a few sample periods for each source, and analyzed all JEM-X and IBIS/ISGRI data with offsets <4 degrees. We seek an explanation for the dichotomy between the hard X-ray tails or lack thereof in the (otherwise very similar) X-ray spectra of Z sources and bright Atolls, respectively.

  2. The power-law and linear correlations between the twin kHz QPO frequencies in LMXBs

    NASA Astrophysics Data System (ADS)

    Yin, H. X.; Zhao, Y. H.

    After the launch of the Rossi X-ray Timing Explorer RXTE our knowledge of the aperiodic variability of newtron star NS low mass X-ray binaries LMXBs took a substantial step forward especially initiated by the discovery of the kilohertz quasi-periodic oscillations kHz QPOs in about twenty more NS LMXBs We analyzed the recently updated kHz QPO data in NS LMXBs in order to investigate the different correlations of the twin peak kilohertz quasi-periodic oscillations kHz QPOs in bright Z sources and in the less luminous Atoll sources We fitted the data with a power-law relation nu 1 sim nu 2 b and a linear relation nu 2 sim A nu 1 between the upper and the lower kHz QPOs and find that both relations can fit the data well with small chi 2 values sim 1 The implications of our results for the theoretical models for kHz QPOs are discussed

  3. X-ray variability with spectral state transitions in NS-LMXBs observed with MAXI/GSC and Swift/BAT

    NASA Astrophysics Data System (ADS)

    Asai, Kazumi; Mihara, Tatehiro; Matsuoka, Masaru; Sugizaki, Mutsumi

    2015-10-01

    X-ray variabilities with spectral state transitions in bright low-mass X-ray binaries containing a neutron star are investigated by using the one-day bin light curves of MAXI/GSC (Gas Slit Camera) and Swift/BAT (Burst Alert Telescope). Four sources (4U 1636-536, 4U 1705-44, 4U 1608-52, and GS 1826-238) exhibited small-amplitude X-ray variabilities with spectral state transitions. Such "mini-outbursts" were characterized by smaller amplitudes (several times) and shorter duration (less than several tens of days) than those of "normal outbursts." A theoretical model of disk instability by Mineshige and Osaki (PASJ, 37, 1, 1985) predicts both large-amplitude outbursts and small-amplitude variabilities. We interpret the normal outbursts as the former prediction of this model, and the mini-outbursts as the latter. Here, we can also call the mini-outburst a "purr-type outburst" referring to the theoretical work. We suggest that similar variabilities lasting for several tens of days without spectral state transitions, which are often observed in the hard state, may be repeats of mini-outbursts.

  4. The reflection component in NS LMXBs

    NASA Astrophysics Data System (ADS)

    D'Aí, A.; Papitto, A.; Di Salvo, T.; Iaria, R.; Robba, N. R.; Egron, E.; Piraino, S.

    2014-01-01

    Thanks to the good spectral resolution and large effective area of the EPIC/PN instrument on board of XMM-Newton, we have at hand a large number of observations of accreting low-mass X-ray binaries, that allow for the fist time a comprehensive view on the characteristics of the reflection component at different accretion regimes and to probe the effects of a magnetosphere on its formation. We focus here on a comparative analysis of the reflection component from a series of spectroscopic studies on selected sources: 4U 1705-44, observed both in the soft and hard state, the pulsating ms pulsars SAX J1808.4-3658 and IGR J17511-3057, and the intermittent pulsar HETE J1900-2455. Although the sources can present very similar accretion rates and continuum shapes, the reflection parameters do not generally result the same, moreover the effect of a magnetosphere on the formation of the reflection component appears elusive.

  5. Boundary layer emission in luminous LMXBs

    NASA Astrophysics Data System (ADS)

    Gilfanov, M.; Revnivtsev, M.

    We show that aperiodic and quasiperiodic variability of bright LMXBs -- atoll and Z- sources on sim sec -- msec time scales is caused primarily by variations of the luminosity of the boundary layer The kHz QPOs have the same origin as variability at lower frequencies i e independent of the nature of the clock the actual luminosity modulation takes place on the neutron star surface The boundary layer spectrum remains nearly constant in the course of the luminosity variations and is represented to certain accuracy by the Fourier frequency resolved spectrum In the investigated range of dot M sim 0 1-1 dot MEdd it depends weakly on the global mass accretion rate and in the limit dot M sim dot MEdd is close to Wien spectrum with kT sim 2 4 keV Its independence on the global value of dot M lends support to the theoretical suggestion by Inogamov Sunyaev 1999 that the boundary layer is radiation pressure supported Based on the knowledge of the boundary layer spectrum we attempt to relate the motion along the Z-track to changes of physically meaningful parameters Our results suggest that the contribution of the boundary layer to the observed emission decreases along the Z-track from conventional sim 50 on the horizontal branch to a rather small number on the normal branch This decrease can be caused for example by obscuration of the boundary layer by the geometrically thickened accretion disk at dot M sim dot MEdd Alternatively this can indicate significant change of

  6. Dust Modeling of Si K Absorption in Galactic Bulge LMXBs with Chandra

    NASA Astrophysics Data System (ADS)

    Schulz, Norbert S.; Corrales, Lia; Canizares, Claude R.

    2016-04-01

    The Galactic Bulge hosts a large number of bright and highly absorbed low-mass X-ray binaries (LMXBs). Column densitiesbetween 1022 cm-2 and 5x1023 cm-2 offer the opportunity and contrast to study the Si K edge structure with very high spectral resolution. Recent models predict that the total extinction in X-ray spectra not only involves X-ray absorption from gas and dust along the line of sight, but also significant contributions from dust scattering. A survey with the Chandra HETG of about a dozen LMXBs yields a rich variety of spectral features, showing that the Si K edge structure is highly complex and variable, from source to source and with time for a given source. We find substructure from neutral atomic silicon, silicate dust absorption and scattering from the interstellar medium (ISM), and superimposed ionized absorption signatures from the circumstellar environment of the LMXBs.

  7. Boundary layer emission in luminous LMXBs

    NASA Astrophysics Data System (ADS)

    Gilfanov, M.; Revnivtsev, M.

    2005-11-01

    We show that aperiodic and quasiperiodic variability of bright LMXBs - atoll and Z-sources - on ˜ sec-msec time scales is caused primarily by variations of the luminosity of the boundary layer. The emission of the accretion disk is less variable on these time scales and its power density spectrum follows P_disk(f)∝ f-1 law, contributing to observed flux variation at low frequencies and low energies only. The kHz QPOs have the same origin as variability at lower frequencies, i.e. independent of the nature of the "clock", the actual luminosity modulation takes place on the neutron star surface. The boundary layer spectrum remains nearly constant in the course of the luminosity variations and is represented to certain accuracy by the Fourier frequency resolved spectrum. In the investigated range of \\dot{M}˜ (0.1-1) \\dot{M}_Edd it depends weakly on the global mass accretion rate and in the limit \\dot{M}˜ \\dot{M}_Edd is close to Wien spectrum with kT˜ 2.4 keV. Its independence on the global value of \\dot{M} lends support to the theoretical suggestion by \\citet{inogamov99} that the boundary layer is radiation pressure supported. \\ Based on the knowledge of the boundary layer spectrum we attempt to relate the motion along the Z-track to changes of physically meaningful parameters. Our results suggest that the contribution of the boundary layer to the observed emission decreases along the Z-track from conventional ˜ 50% on the horizontal branch to a rather small number on the normal branch. This decrease can be caused, for example, by obscuration of the boundary layer by the geometrically thickened accretion disk at \\dot{M}˜\\dot{M}_Edd. Alternatively, this can indicate significant change of the structure of the accretion flow at \\dot{M}˜\\dot{M}_Edd and disappearance of the boundary layer as a distinct region of the significant energy release associated with the neutron star surface.

  8. A new evolutionary picture for CVs and LMXBs

    NASA Astrophysics Data System (ADS)

    King, A. R.; Schenker, K.

    2002-01-01

    We consider an alternative to the standard picture of CV and LMXB evolution, namely the idea that most CVs (and by extension LMXBs) may not yet have had time to evolve to their theoretical minimum orbital periods. We call this the Binary Age Postulate (BAP). The observed short--period cutoff in the CV histogram emerges naturally as the shortest period yet reached in the age of the Galaxy, while the post--minimum--period space density problem is removed. The idea has similar desirable consequences for LMXBs. In both cases systems with nuclear--evolved secondary stars form a prominent part of the short--period distributions. Properties such as the existence and nature of ultrashort--period systems, and the spread in mass transfer rates at a given orbital period, are naturally reproduced.

  9. Extended Corbet Diagram of HMXBs, LMXBs and radio pulsar binaries

    NASA Astrophysics Data System (ADS)

    Yan, Yan; Ali, Esamdin; Yin, Hongxing

    2010-09-01

    The evolutionary scenario of neutron star binaries is still an essential enigma in both stellar astrophysics and high energy astrophysics. In order to explore the scenario, we include the accumulation of data on the orbits and spins of compact binaries in multi-wavelength ranging from radio to X-ray, such as radio pulsar binaries, HMXBs, and LMXBs, filling them into the so called “Corbet Diagram” which initially investigated the period of orbit ( P orb)˜ the period of spin ( P spin) correlation of HMXBs. We find that the evolutionary scenario comes more clearly and makes strong confirmation of the connection between LMXBs and radio pulsar binaries, predicted by the recycle process. However, the origins of radio pulsar binaries sre still unknown. Accretion Induced Collapse (AIP) process may be a mechanism which can explain the origin of the binary millisecond pulsars with relatively longer orbital periods. A correlation of P {orb/1/3} ˜ P {spin/-1} of LMXBs and radio pulsar binaries may exist.

  10. Multi-wavelength coverage of outburst decays of LMXBs

    NASA Astrophysics Data System (ADS)

    Kalemci, Emrah; Tomsick, John; Bailyn, Charles; Dincer, Tolga

    2016-07-01

    Low mass X-ray binaries (LMXB) are dynamic laboratories that are powered by accretion, and under some conditions these systems create strong outflows in the form of highly collimated jets, or winds. Outburst decays of transient LMXBs provide additional information about compact jets and their relation to changes in timing properties. Our group characterizes the multi-wavelength evolution of Galactic black hole transients during their outburst decays using simultaneous X-ray (RXTE, Swift, and INTEGRAL), optical/infrared (SMARTS) and radio (VLA, ATCA, VLBI). By characterizing the X-ray spectral and timing evolution of these systems, and merging this information with the evolution in the near-infrared and radio, we obtain the conditions necessary to launch stable compact jets, and discuss how jets can be influencing and/or influenced by X-ray timing and spectral properties of these systems. In this presentation, I will summarize recent results regarding multi-wavelength observations of not only black hole systems, but also neutron star X-ray binaries during outburst decays, and discuss models that explain not only the broad spectral energy distribution, but also some of the timing properties of these systems.

  11. VizieR Online Data Catalog: LMXBs detected in nearby galaxies (Zhang+, 2011)

    NASA Astrophysics Data System (ADS)

    Zhang, Z.; Gilfanov, M.; Voss, R.; Sivakoff, G. R.; Kraft, R. P.; Brassington, N. J.; Kundu, A.; Jordan, A.; Sarazin, C.

    2011-11-01

    Based on the archival data from the Chandra observations of nearby galaxies, we study different sub populations of low-mass X-ray binaries (LMXBs) - dynamically formed systems in globular clusters (GCs) and in the nucleus of M 31 and (presumably primordial) X-ray binaries in the fields of galaxies. Our aim is to produce accurate luminosity distributions of X-ray binaries in different environments, suitable for quantitative comparison with each other and with the output of population synthesis calculations. Our sample includes seven nearby galaxies (M 31, Maffei 1, Centaurus A, M 81, NGC 3379, NGC 4697, and NGC 4278) and the Milky Way, which together provide relatively uniform coverage down to the luminosity limit of 1035erg/s. In total we have detected 185 LMXBs associated with GCs, 35 X-ray sources in the nucleus of M 31, and 998 field sources of which ~365 are expected to be background AGN. We combine these data, taking special care to accurately account for X-ray and optical incompleteness corrections and the removal of the contamination from the cosmic X-ray background sources, to produce luminosity distributions of X-ray binaries in different environments to far greater accuracy than has been obtained previously. (2 data files).

  12. Modeling the Spin Equilibrium of Neutron Stars in LMXBs Without Gravitational Radiation

    NASA Technical Reports Server (NTRS)

    Andersson, N.; Glampedakis, K.; Haskell, B.; Watts, A. L.

    2004-01-01

    In this paper we discuss the spin-equilibrium of accreting neutron stars in LMXBs. We demonstrate that, when combined with a naive spin-up torque, the observed data leads to inferred magnetic fields which are at variance with those of galactic millisecond radiopulsars. This indicates the need for either additional spin-down torques (eg. gravitational radiation) or an improved accretion model. We show that a simple consistent accretion model can be arrived at by accounting for radiation pressure in rapidly accreting systems (above a few percent of the Eddington accretion rate). In our model the inner disk region is thick and significantly sub-Keplerian, and the estimated equilibrium periods are such that the LMXB neutron stars have properties that accord well with the galactic millisecond radiopulsar sample. The implications for future gravitational-wave observations are also discussed briefly.

  13. Rainbow brightness.

    PubMed

    Gedzelman, S D

    1982-08-15

    A theory for the brightness of rainbows is presented. The light reaching the observer consists of a beam of singly scattered sunlight, originating from the directly illuminated portion of a rainswath, which, in turn, has suffered depletion by scattering or absorption in its path through the atmosphere. The model incorporates the relevant features of cloud geometry and solar position in relation to the observer appropriate to rainbows. The model helps explain why the bottom (or near-horizon portion) of the rainbow tends to be both brighter and redder than the top (or horizontal portion furthest above the ground) when the sun is near the horizon. The greater brightness of the bottom of the bow derives principally from the greater length of the directly illuminated part of the rainswath near the horizon, while the increased redness of the bow's bottom is due to the severe depletion of the short-wavelength contribution to the rainbow beam in its passage through the atmosphere. PMID:20396168

  14. In vitro processing of dengue virus type 2 nonstructural proteins NS2A, NS2B, and NS3.

    PubMed Central

    Preugschat, F; Yao, C W; Strauss, J H

    1990-01-01

    We have tested the hypothesis that the flavivirus nonstructural protein NS3 is a viral proteinase that generates the termini of several nonstructural proteins by using an efficient in vitro expression system and monospecific antisera directed against the nonstructural proteins NS2B and NS3. A series of cDNA constructs was transcribed by using T7 RNA polymerase, and the RNA was translated in reticulocyte lysates. The resulting protein patterns indicated that proteolytic processing occurred in vitro to generate NS2B and NS3. The amino termini of NS2B and NS3 produced in vitro were found to be the same as the termini of NS2B and NS3 isolated from infected cells. Deletion analysis of cDNA constructs localized the protease domain within NS3 to the first 184 amino acids but did not eliminate the possibility that sequences within NS2B were also required for proper cleavage. Kinetic analysis of processing events in vitro and experiments to examine the sensitivity of processing to dilution suggested that an intramolecular cleavage between NS2A and NS2B preceded an intramolecular cleavage between NS2B and NS3. The data from these expression experiments confirm that NS3 is the viral proteinase responsible for cleavage events generating the amino termini of NS2B and NS3 and presumably for cleavages generating the termini of NS4A and NS5 as well. Images PMID:2143543

  15. BrightFocus Foundation

    MedlinePlus

    ... Alzheimer’s Disease Research Program Macular Degeneration Research Program National Glaucoma Research Program Molecular Neurodegeneration ... Foundation BrightFocus Foundation 22512 Gateway Center Drive Clarksburg, MD ...

  16. Brightness of Moonlight.

    ERIC Educational Resources Information Center

    Garstang, R. H.

    1985-01-01

    Measurement of the brightness of moonlight by comparison with lamp-light from a low wattage light bulb is an elementary project in astronomy which illustrates scientific principles for the freshman level. Two methods used for the comparison (shadow brightness method and grease spot method) are explained, with suggestions and expected answers. (DH)

  17. Star Light, Star Bright.

    ERIC Educational Resources Information Center

    Iadevaia, David G.

    1984-01-01

    Presents a technique for obtaining a rough measure of the brightness among different stars. Materials needed include a standard 35-mm camera, a plastic ruler, and a photo enlarger. Although a telescope can be used, it is not essential. (JN)

  18. NS&T Management Observations

    SciTech Connect

    Gianotto, David

    2014-09-01

    The INL Management Observation Program (MOP) is designed to improve managers and supervisors understanding of work being performed by employees and the barriers impacting their success. The MOP also increases workers understanding of managements’ expectations as they relate to safety, security, quality, and work performance. Management observations (observations) are designed to improve the relationship and trust between employees and managers through increased engagement and interactions between managers and researchers in the field. As part of continuous improvement, NS&T management took initiative to focus on the participation and quality of observations in FY 14. This quarterly report is intended to (a) summarize the participation and quality of management’s observations, (b) assess observations for commonalities or trends related to facility or process barriers impacting research, and (c) provide feedback and make recommendations for improvements NS&T’s MOP.

  19. Bright patches on Ariel

    NASA Technical Reports Server (NTRS)

    1986-01-01

    Distinct bright patches are visible on Ariel, the brightest of Uranus' five largest satellites. Voyager 2 obtained this image Jan. 22, 1986, from a distance of 2.52 million kilometers (1.56 million miles). The clear-filter image, obtained with the narrow-angle camera, shows a resolution of 47 km (29 miles). Ariel is about 1,300 km (800 mi) in diameter. This image shows several distinct bright areas that reflect nearly 45 percent of the incident sunlight; on average, the satellite displays a reflectivity of about 25-30 percent. The bright areas are probably fresh water ice, perhaps excavated by impacts. The south pole of Ariel is slightly off center of the disk in this view. Voyager 2 will obtain its best views of the satellite on Jan. 24, at a closest-approach distance of 127,000 km (79,000 mi). The Voyager project is managed for NASA by the Jet Propulsion Laboratory.

  20. High Brightness Test Stand

    SciTech Connect

    Birx, D.L.; Caporaso, G.J.; Boyd, J.K.; Hawkins, S.A.; Poor, S.E.; Reginato, L.L.; Rogers, D. Jr.; Smith, M.W.

    1985-08-07

    The High Brightness Test Stand is a 2 MeV, less than or equal to 10 kA electron accelerator module. This accelerator module, designed as an upgrade prototype for the Advanced Test Accelerator (ATA), combines solid state nonlinear magnetic drives with state-of-the-art induction linac technology. The facility serves a dual role, as it not only provides a test bed for this new technology, but is used to develop high brightness electron optics. We will both further describe the accelerator, as well as present some of the preliminary electron optics measurements.

  1. Maximum voltage gradient, uniformity and brightness of dispenser cathodes

    SciTech Connect

    Turner, W.C.; Chen, Y.J.; Green, M.C.; Miram, G.V.; Nordquist, A.L.

    1988-01-01

    Initial measurements of current uniformity and intrinsic cathode brightness are presented a 5.1 cm diameter osmium alloy coated dispenser cathode. The data are obtained with a planar diode driven with a 35ns Blumlein pulse line. Pepper pot images are observed with a fast phosphor and gated camera. The current density and beamlet divergence angle are uniform within +-10% and a brightness J = 1.6E10A/m/sup 2/rad/sup 2/ is observed at a current density j=44A/cm/sup 2/. 6 refs., 2 figs., 2 tabs.

  2. Brightness predictions for comets

    NASA Astrophysics Data System (ADS)

    Green, Daniel W. E.; Marsden, Brian G.; Morris, Charles S.

    2001-02-01

    Daniel W E Green, Brian G Marsden and Charles S Morris write with the aim of illuminating the issue of cometary light curves and brightness predictions, following the publication in this journal last October of the letter by John McFarland (2000).

  3. A Bright Shining Lesson

    ERIC Educational Resources Information Center

    Hurowitz, Amanda

    2010-01-01

    Sometimes students come up with crazy ideas. When this author first started teaching at Thomas Jefferson High School for Science and Technology in Virginia five years ago, she had a sophomore share such an idea with her. He wanted to put solar panels on the school's roof as a way to reduce the school's carbon footprint and set a bright clean…

  4. Bright Fireball Over Georgia

    NASA Video Gallery

    A camera in Cartersville, Ga., captured this view of a bright fireball over Georgia on the night of Mar. 7, 2012, at approx. 10:19:11 EST. The meteor was first recorded at an altitude of 51.5 miles...

  5. Bright Streak on Amalthea

    NASA Technical Reports Server (NTRS)

    2000-01-01

    These two images of Jupiter's small, irregularly shaped moon Amalthea, obtained by the camera onboard NASA's Galileo spacecraft in August 1999(left) and November 1999 (right), form a 'stereo pair' that helps scientists determine this moon's shape and the topography of its surface features. Features as small as 3.8 kilometers (2.4 miles) across can be resolved in these images, making them among the highest-resolution images ever taken of Amalthea.

    The large impact crater visible in both images, near the right-hand edge of Amalthea's disk, is about 40 kilometers (about 29 miles) across; two ridges, tall enough to cast shadows, extend from the top of the crater in a V-shape reminiscent of a 'rabbit ears' television antenna. To the left of these ridges, in the top center portion of Amalthea's disk, is a second large impact crater similar in size to the first crater. To the left of this second crater is a linear 'streak' of relatively bright material about 50 kilometers (31 miles) long. In previous spacecraft images of Amalthea taken from other viewing directions, this bright feature was thought to be a small, round, bright 'spot' and was given the name Ida. These new images reveal for the first time that Ida is actually a long, linear 'streak.' This bright streak may represent material ejected during the formation of the adjacent impact crater, or it may just mark the crest of a local ridge. Other patches of relatively bright material can be seen elsewhere on Amalthea's disk, although none of these other bright spots has Ida's linear shape.

    In both images, sunlight is coming from the left and north is approximately up. Note that the north pole of Amalthea is missing in the right-hand image (it was cut off by the edge of the camera frame). The bright streak, Ida, is on the side of the moon that faces permanently away from Jupiter, and the crater near the right-hand edge of the disk is in the center of Amalthea's leading side (the side of the moon that 'leads

  6. Bright field illumination system

    NASA Technical Reports Server (NTRS)

    Huber, Edward D. (Inventor)

    1998-01-01

    A Bright Field Illumination system for inspecting a range of characteristically different kinds of defects, depressions, and ridges in a selected material surface. The system has an illumination source placed near a first focus of an elliptical reflector. In addition, a camera facing the inspected area is placed near the illumination source and the first focus. The second focus of the elliptical reflector is located at a distance approximately twice the elliptical reflector's distance above the inspected surface. The elliptical reflector directs the light from the source onto the inspected surface. Due to the shape of the elliptical reflector, light that is specularly reflected from the inspected surface is directed into the camera is which located at the position of the reflected second focus of the ellipse. This system creates a brightly lighted background field against which damage sites appear as high contrast dark objects which can be easily detected by a person or an automated inspection system. In addition, the Bright Field Illumination system and method can be used in combination with a vision inspection system providing for multiplexed illumination and data handling of multiple kinds of surface characteristics including abrupt and gradual surface variations and differences between measured characteristics of different kinds and prior instruments.

  7. Kiloamp high-brightness beams

    SciTech Connect

    Caporaso, G.J.

    1987-01-01

    Brightness preservation of high-current relativistic electron beams under two different types of transport is discussed. Recent progress in improving the brightness of laser-guided beams in the Advanced Test Accelerator is reviewed. A strategy for the preservation of the brightness of space-charge-dominated beams in a solenoidal transport system is presented.

  8. Low surface brightness galaxies

    NASA Technical Reports Server (NTRS)

    Vanderhulst, J. M.; Deblok, W. J. G.; Mcgaugh, S. S.; Bothun, G. D.

    1993-01-01

    A program to investigate the properties of low surface brightness (LSB) galaxies involving surface photometry in U, B, V, R, I, and H-alpha, HI imaging with the Westerbork Synthesis Radio Telescope (WSRT) and the very large array (VLA) and spectrophotometry of H2 regions in LSB galaxies is underway. The goal is to verify the idea that LSB galaxies have low star formation rates because the local gas density falls below the critical density for star formation, and to study the stellar population and abundances in LSB galaxies. Such information should help understanding the evolutionary history of LSB galaxies. Some preliminary results are reported.

  9. Small bright charged colloidal quantum dots.

    PubMed

    Qin, Wei; Liu, Heng; Guyot-Sionnest, Philippe

    2014-01-28

    Using electrochemical charge injection, the fluorescence lifetimes of negatively charged core/shell CdTe/CdSe QDs are measured as a function of core size and shell thickness. It is found that the ensemble negative trion lifetimes reach a maximum (∼4.5 ns) for an intermediate shell thickness. This leads to the smallest particles (∼4.5 nm) with the brightest trion to date. Single dot measurements show that the negative charge suppresses blinking and that the trion can be as bright as the exciton at room temperature. In contrast, the biexciton lifetimes remain short and exhibit only a monotonous increase with shell thickness, showing no correlation with the negative trion decays. The suppression of the Auger process in small negatively charged CdTe/CdSe quantum dots is unprecedented and a significant departure from prior results with ultrathick CdSe/CdS core/shell or dot-in-rod structures. The proposed reason for the optimum shell thickness is that the electron-hole overlap is restricted to the CdTe core while the electron is tuned to have zero kinetic energy in the core for that optimum shell thickness. The different trend of the biexciton lifetime is not explained but tentatively attributed to shorter-lived positive trions at smaller sizes. These results improve our understanding of multiexciton recombination in colloidal quantum dots and may lead to the design of bright charged QDs for more efficient light-emitting devices.

  10. Monoclonal Antibodies Against NS3 and NS5 Proteins of Japanese Encephalitis Virus

    PubMed Central

    Chen, Zheng; Shao, Lin; Ye, Jing; Li, Yongmao; Huang, Shaomei; Chen, Huanchun

    2012-01-01

    Non-structural proteins NS3 and NS5 of Japanese encephalitis virus (JEV) were expressed in Escherichia coli and purified by dialysis. Two monoclonal antibodies (MAbs) named 1H7 and 2D4 against NS3 protein and three MAbs named 3C4, 3H7, and 3F10 against NS5 protein were generated by fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from NS3 or NS5 protein immunized mice. Then activity of MAbs was characterized by enzyme-linked immunosorbent assay (ELISA), Western blot analysis, and indirect immunofluorescent assays (IFA). Our results demonstrated that all the MAbs showed high specificity and sensitivity in IFA at 1:100 dilution and in Western blot analysis at 1:500 dilution, which indicated that these MAbs against NS3 and NS5 proteins of JEV may be used as valuable tools for analysis of the protein functions and pathogenesis of JEV. PMID:22509919

  11. High brightness electron sources

    SciTech Connect

    Sheffield, R.L.

    1995-07-01

    High energy physics accelerators and free electron lasers put increased demands on the electron beam sources. This paper describes the present research on attaining intense bright electron beams using photoinjectors. Recent results from the experimental programs will be given. The performance advantages and difficulties presently faced by researchers will be discussed, and the following topics will be covered. Progress has been made in photocathode materials, both in lifetime and quantum efficiency. Cesium telluride has demonstrated significantly longer lifetimes than cesium antimonide at 10{sup {minus}8} torr. However, the laser system is more difficult because cesium telluride requires quadrupled YLF instead of the doubled YLF required for cesium antimonide. The difficulty in using photoinjectors is primarily the drive laser, in particular the amplitude stability. Finally, emittance measurements of photoinjector systems can be complicated by the non-thermal nature of the electron beam. An example of the difficulty in measuring beam emittance is given.

  12. High brightness electron accelerator

    DOEpatents

    Sheffield, Richard L.; Carlsten, Bruce E.; Young, Lloyd M.

    1994-01-01

    A compact high brightness linear accelerator is provided for use, e.g., in a free electron laser. The accelerator has a first plurality of acclerating cavities having end walls with four coupling slots for accelerating electrons to high velocities in the absence of quadrupole fields. A second plurality of cavities receives the high velocity electrons for further acceleration, where each of the second cavities has end walls with two coupling slots for acceleration in the absence of dipole fields. The accelerator also includes a first cavity with an extended length to provide for phase matching the electron beam along the accelerating cavities. A solenoid is provided about the photocathode that emits the electons, where the solenoid is configured to provide a substantially uniform magnetic field over the photocathode surface to minimize emittance of the electons as the electrons enter the first cavity.

  13. Classical swine fever virus NS5B protein suppresses the inhibitory effect of NS5A on viral translation by binding to NS5A.

    PubMed

    Sheng, Chun; Wang, Jing; Xiao, Jing; Xiao, Jun; Chen, Yan; Jia, Lin; Zhi, Yimiao; Li, Guangyuan; Xiao, Ming

    2012-05-01

    In order to investigate molecular mechanisms of internal ribosome entry site (IRES)-mediated translation in classical swine fever virus (CSFV), an important pathogen of pigs, the expression level of NS3 was evaluated in the context of genomic RNAs and reporter RNA fragments. All data showed that the NS5A protein has an inhibitory effect on IRES-mediated translation and that NS5B proteins suppress the inhibitory effect of NS5A on viral translation, but CSFV NS5B GDD mutants do not. Furthermore, glutathione S-transferase pull-down assay and immunoprecipitation analysis, associated with deletion and alanine-scanning mutations, were performed. Results showed that NS5B interacts with NS5A and that the region aa 390-414, located in the C-terminal half of NS5A, is important for binding of NS5B to NS5A. Furthermore, amino acids K399, T401, E406 and L413 in the region were found to be essential for NS5A-NS5B interaction, virus rescue and infection. The above-mentioned region and four amino acids were observed to overlap with the site responsible for inhibition of IRES-mediated translation by the NS5A protein. We also found that aa 63-72, aa 637-653 and the GDD motif of NS5B were necessary for the interaction between NS5A and NS5B. These findings suggest that the repression activity of the NS5B protein toward the role of NS5A in translation might be achieved by NS5A-NS5B interaction, for which aa 390-414 of NS5A and aa 63-72, aa 637-653 and the GDD motif of NS5B are indispensable. This is important for understanding the role of NS5A-NS5B interaction in the virus life cycle. PMID:22258858

  14. Dengue 2 virus NS2B and NS3 form a stable complex that can cleave NS3 within the helicase domain.

    PubMed

    Arias, C F; Preugschat, F; Strauss, J H

    1993-04-01

    Flavivirus genomic RNA is translated into a large polyprotein that is processed into structural and nonstructural proteins. The N-termini of several nonstructural proteins are produced by cleavage at dibasic sites by a two-component viral proteinase consisting of NS2B and NS3. NS3 contains a trypsin-like serine proteinase domain at its N-terminus, whereas the function of NS2B in proteolysis is yet to be determined. We have used an NS3-specific antiserum, under nondenaturing conditions, to demonstrate that NS2B and NS3 form a complex both in vitro and in vivo. The N-terminal 184 residues of NS3 are sufficient to form the complex with NS2B. The complex forms efficiently when the NS2B and NS3 are translated from two different mRNAs as well as when NS2B and NS3 are translated as a polyprotein from the same mRNA. A chimeric complex can be formed between yellow fever NS2B and a chimeric yellow fever-dengue 2 NS3. Using anti-NS3 antisera, we also found that a 50-kDa fragment of NS3, consisting of the N-terminal approximately 460 residues, is produced in infected mammalian cells. This fragment is not produced in infected mosquito cells, but will form in Triton X-100 lysates of mosquito cells. The cleavage of NS3 to form this fragment is catalyzed by the NS3 proteinase itself and proteolysis requires NS2B. Examination of the amino acid sequence of NS3 reveals a potential conserved cleavage site that resembles other sites cleaved by the NS3/NS2B proteinase; this site occurs within a conserved RNA helicase sequence motif. The importance of this alternatively processed form of NS3 and its role in the replication cycle of dengue virus remain to be determined.

  15. Allosteric inhibition of the NS2B-NS3 protease from dengue virus.

    PubMed

    Yildiz, Muslum; Ghosh, Sumana; Bell, Jeffrey A; Sherman, Woody; Hardy, Jeanne A

    2013-12-20

    Dengue virus is the flavivirus that causes dengue fever, dengue hemorrhagic disease, and dengue shock syndrome, which are currently increasing in incidence worldwide. Dengue virus protease (NS2B-NS3pro) is essential for dengue virus infection and is thus a target of therapeutic interest. To date, attention has focused on developing active-site inhibitors of NS2B-NS3pro. The flat and charged nature of the NS2B-NS3pro active site may contribute to difficulties in developing inhibitors and suggests that a strategy of identifying allosteric sites may be useful. We report an approach that allowed us to scan the NS2B-NS3pro surface by cysteine mutagenesis and use cysteine reactive probes to identify regions of the protein that are susceptible to allosteric inhibition. This method identified a new allosteric site utilizing a circumscribed panel of just eight cysteine variants and only five cysteine reactive probes. The allosterically sensitive site is centered at Ala125, between the 120s loop and the 150s loop. The crystal structures of WT and modified NS2B-NS3pro demonstrate that the 120s loop is flexible. Our work suggests that binding at this site prevents a conformational rearrangement of the NS2B region of the protein, which is required for activation. Preventing this movement locks the protein into the open, inactive conformation, suggesting that this site may be useful in the future development of therapeutic allosteric inhibitors. PMID:24164286

  16. The C-terminal 50 amino acid residues of dengue NS3 protein are important for NS3-NS5 interaction and viral replication.

    PubMed

    Tay, Moon Y F; Saw, Wuan Geok; Zhao, Yongqian; Chan, Kitti W K; Singh, Daljit; Chong, Yuwen; Forwood, Jade K; Ooi, Eng Eong; Grüber, Gerhard; Lescar, Julien; Luo, Dahai; Vasudevan, Subhash G

    2015-01-23

    Dengue virus multifunctional proteins NS3 protease/helicase and NS5 methyltransferase/RNA-dependent RNA polymerase form part of the viral replication complex and are involved in viral RNA genome synthesis, methylation of the 5'-cap of viral genome, and polyprotein processing among other activities. Previous studies have shown that NS5 residue Lys-330 is required for interaction between NS3 and NS5. Here, we show by competitive NS3-NS5 interaction ELISA that the NS3 peptide spanning residues 566-585 disrupts NS3-NS5 interaction but not the null-peptide bearing the N570A mutation. Small angle x-ray scattering study on NS3(172-618) helicase and covalently linked NS3(172-618)-NS5(320-341) reveals a rigid and compact formation of the latter, indicating that peptide NS5(320-341) engages in specific and discrete interaction with NS3. Significantly, NS3:Asn-570 to alanine mutation introduced into an infectious DENV2 cDNA clone did not yield detectable virus by plaque assay even though intracellular double-stranded RNA was detected by immunofluorescence. Detection of increased negative-strand RNA synthesis by real time RT-PCR for the NS3:N570A mutant suggests that NS3-NS5 interaction plays an important role in the balanced synthesis of positive- and negative-strand RNA for robust viral replication. Dengue virus infection has become a global concern, and the lack of safe vaccines or antiviral treatments urgently needs to be addressed. NS3 and NS5 are highly conserved among the four serotypes, and the protein sequence around the pinpointed amino acids from the NS3 and NS5 regions are also conserved. The identification of the functionally essential interaction between the two proteins by biochemical and reverse genetics methods paves the way for rational drug design efforts to inhibit viral RNA synthesis.

  17. A high brightness photoinjector

    NASA Astrophysics Data System (ADS)

    Le Sage, Gregory Peter

    Linear colliders, future electron acceleration schemes, and short pulse, ultrawideband millimeter-wave sources require very bright electron beams. Conventional electron injectors including thermionic cathodes and RF bunchers or DC guns have intrinsic limitations which preclude their usage for many of these applications. RF photoinjectors have shown their ability to produce relativistic electron beams with low emittance and energy spread. However, previously developed RF photoinjectors are also subject to significant limitations. These include extreme sensitivity to timing between the RF in the accelerator structure and the drive laser, low efficiency with respect to the number and charge of the electron bunches produced by the injector, and high cost associated with both the RF drive and laser systems. The presently described system has addressed these issues by combining state-of-the-art capabilities in the laser and RF systems, photocathode materials, and new concepts for synchronization. Phase jitter generated by sources including Klystron modulator voltage fluctuation has been measured in detail, and schemes for alleviating this problem have undergone initial proof-of-principle testing. New concepts for the drive laser system have been tested which will lead to further improvements in performance, simplicity, cost-effectiveness, and compactness. The analytical and experimental work associated with the development of a high brightness, high gradient electron accelerator is presented. The presentation emphasizes the systematic progress toward the original design goals of the project, as well as the state-of-the-art innovations characterizing the system. The linear electron accelerator system is based on a 1 1/2 cell side-wall coupled, π-mode standing wave accelerator structure, driven by a 20 MW SLAC Klystron operating at 8.548 GHz, a Ti:Sapphire laser oscillator, and an 8-pass, chirped pulse Ti:Sapphire laser amplifier. Simulations show an rms transverse

  18. How Bright Is the Sun?

    ERIC Educational Resources Information Center

    Berr, Stephen

    1991-01-01

    Presents a sequence of activities designed to allow eighth grade students to deal with one of the fundamental relationships that govern energy distribution. Activities guide students to measure light bulb brightness, discover the inverse square law, compare light bulb light to candle light, and measure sun brightness. (two references) (MCO)

  19. Spin Complicates Eccentric BH-NS Mergers

    NASA Astrophysics Data System (ADS)

    Kohler, Susanna

    2015-08-01

    When a neutron star (NS) has a glancing encounter with a black hole (BH), its spin has a significant effect on the outcome, according to new simulations run by William East of Stanford University and his collaborators. Spotting an Eccentric Merger. In a traditional BH-NS merger, the two objects orbit each other quasi-circularly as they spiral in. But there's another kind of merger that's possible in high-density environments like galactic nuclei or globular clusters: a dynamical capture merger, in which a NS and BH pass each other just close enough that the gravity of the black hole "catches" the NS, leading the two objects to merge with very eccentric orbits. During an eccentric merger, the NS can be torn apart -- at which point some fraction of the tidally-disrupted material will escape the system, while some fraction instead accretes back onto the BH. Knowing these fractions is important for being able to model the expected electromagnetic signatures for the merger: the unbound material can power transients like kilonovae, whereas the accreting material may be the cause of short gamma-ray bursts. The amount of material available for events like these would change their observable strengths. Testing the Effects of Spin. To see whether NS spin has an impact on the behavior of the merger, East and collaborators use a general-relativistic hydrodynamic code to simulate the glancing encounter of a BH and a NS with dimensionless spin between a=0 (non-spinning) and a=0.756 (rotation period of 1 ms). They also vary the separation of the first encounter. The group finds that changing the NS's spin can change a number of outcomes of the merger. To start with, it can affect whether the NS is captured by the BH, or if the encounter is glancing and then both objects carry on their merry way. And if the NS is trapped by the BH and torn apart, then the higher the NS's spin, the more matter outside of the BH ends up unbound, instead of getting trapped into an accretion disk

  20. Rationalizing meat consumption. The 4Ns.

    PubMed

    Piazza, Jared; Ruby, Matthew B; Loughnan, Steve; Luong, Mischel; Kulik, Juliana; Watkins, Hanne M; Seigerman, Mirra

    2015-08-01

    Recent theorizing suggests that the 4Ns - that is, the belief that eating meat is natural, normal, necessary, and nice - are common rationalizations people use to defend their choice of eating meat. However, such theorizing has yet to be subjected to empirical testing. Six studies were conducted on the 4Ns. Studies 1a and 1b demonstrated that the 4N classification captures the vast majority (83%-91%) of justifications people naturally offer in defense of eating meat. In Study 2, individuals who endorsed the 4Ns tended also to objectify (dementalize) animals and included fewer animals in their circle of moral concern, and this was true independent of social dominance orientation. Subsequent studies (Studies 3-5) showed that individuals who endorsed the 4Ns tend not to be motivated by ethical concerns when making food choices, are less involved in animal-welfare advocacy, less driven to restrict animal products from their diet, less proud of their animal-product decisions, tend to endorse Speciesist attitudes, tend to consume meat and animal products more frequently, and are highly committed to eating meat. Furthermore, omnivores who strongly endorsed the 4Ns tended to experience less guilt about their animal-product decisions, highlighting the guilt-alleviating function of the 4Ns. PMID:25865663

  1. Rationalizing meat consumption. The 4Ns.

    PubMed

    Piazza, Jared; Ruby, Matthew B; Loughnan, Steve; Luong, Mischel; Kulik, Juliana; Watkins, Hanne M; Seigerman, Mirra

    2015-08-01

    Recent theorizing suggests that the 4Ns - that is, the belief that eating meat is natural, normal, necessary, and nice - are common rationalizations people use to defend their choice of eating meat. However, such theorizing has yet to be subjected to empirical testing. Six studies were conducted on the 4Ns. Studies 1a and 1b demonstrated that the 4N classification captures the vast majority (83%-91%) of justifications people naturally offer in defense of eating meat. In Study 2, individuals who endorsed the 4Ns tended also to objectify (dementalize) animals and included fewer animals in their circle of moral concern, and this was true independent of social dominance orientation. Subsequent studies (Studies 3-5) showed that individuals who endorsed the 4Ns tend not to be motivated by ethical concerns when making food choices, are less involved in animal-welfare advocacy, less driven to restrict animal products from their diet, less proud of their animal-product decisions, tend to endorse Speciesist attitudes, tend to consume meat and animal products more frequently, and are highly committed to eating meat. Furthermore, omnivores who strongly endorsed the 4Ns tended to experience less guilt about their animal-product decisions, highlighting the guilt-alleviating function of the 4Ns.

  2. Hepatitis C virus core, NS3, NS4B and NS5A are the major immunogenic proteins in humoral immunity in chronic HCV infection

    PubMed Central

    Sillanpää, Maarit; Melén, Krister; Porkka, Päivi; Fagerlund, Riku; Nevalainen, Kaisu; Lappalainen, Maija; Julkunen, Ilkka

    2009-01-01

    Background The viral genome of hepatitis C virus constitutes a 9.6-kb single-stranded positive-sense RNA which encodes altogether 11 viral proteins. In order to study the humoral immune responses against different HCV proteins in patients suffering from chronic HCV infection, we produced three structural (core, E1 and E2) and six nonstructural proteins (NS2, NS3, NS4A, NS4B, NS5A and NS5B) in Sf9 insect cells by using the baculovirus expression system. Results The recombinant HCV core, E1, E2, NS2, NS3, NS4A, NS4B, NS5A and NS5B proteins were purified and used in Western blot analysis to determine antibody responses against individual HCV protein in 68 HCV RNA and antibody positive human sera that were obtained from patients suffering from genotype 1, 2, 3 or 4 infection. These sera were also analysed with INNO-LIA Score test for HCV antibodies against core, NS3, NS4AB and NS5A, and the results were similar to the ones obtained by Western blot method. Based on our Western blot analyses we found that the major immunogenic HCV antigens were the core, NS4B, NS3 and NS5A proteins which were recognized in 97%, 86%, 68% and 53% of patient sera, respectively. There were no major genotype specific differences in antibody responses to individual HCV proteins. A common feature within the studied sera was that all except two sera recognized the core protein in high titers, whereas none of the sera recognized NS2 protein and only three sera (from genotype 3) recognised NS5B. Conclusion The data shows significant variation in the specificity in humoral immunity in chronic HCV patients. PMID:19549310

  3. Nucleocytoplasmic transport: the influenza virus NS1 protein regulates the transport of spliced NS2 mRNA and its precursor NS1 mRNA.

    PubMed

    Alonso-Caplen, F V; Nemeroff, M E; Qiu, Y; Krug, R M

    1992-02-01

    Influenza virus unspliced NS1 mRNA, like retroviral pre-mRNAs, is efficiently exported from the nucleus and translated in the cytoplasm of infected cells. With human immunodeficiency virus (HIV), the transport of viral pre-mRNAs is facilitated by the viral Rev protein. We tested the possibility that the influenza virus NS1 protein, a nuclear protein that is encoded by unspliced NS1 mRNA, has the same function as the HIV Rev protein. Surprisingly, using transient transfection assays, we found that rather than facilitating the nucleocytoplasmic transport of unspliced NS1 mRNA, the NS1 protein inhibited the transport of NS2 mRNA, the spliced mRNA generated from NS1 mRNA. The efficient transport of NS2 mRNA from the nucleus to the cytoplasm occurred only when the synthesis of the NS1 protein was abrogated by amber mutations. The NS1 protein down-regulated the export of NS2 mRNA whether or not it was generated by splicing, indicating that the NS1 protein acted directly on transport. Actinomycin D chase experiments verified that the NS1 protein acted on the transport and not on the differential stability of NS2 mRNA in the nucleus as compared to the cytoplasm. In addition, the NS1 protein inhibited the transport of NS1 mRNA itself, which contains all of the sequences in NS2 mRNA, particularly when NS1 mRNA was released from the splicing machinery by mutating its 3'-splice site. Our results indicate that the NS1 protein-mediated inhibition of transport requires sequences in NS2 mRNA. The transport of the viral PB1 protein, nucleocapsid protein, hemagglutinin, membrane protein, and M2 mRNAs was not affected by the NS1 protein. When the NS2 mRNA sequence was covalently attached to the PB1 mRNA, the transport of the chimeric mRNA was inhibited by the NS1 protein. Our results identify a novel function of the influenza virus NS1 protein and demonstrate that post-transcriptional control of gene expression can also occur at the level of the nucleocytoplasmic transport of a

  4. A novel recombinant single-chain hepatitis C virus NS3-NS4A protein with improved helicase activity.

    PubMed Central

    Howe, A. Y.; Chase, R.; Taremi, S. S.; Risano, C.; Beyer, B.; Malcolm, B.; Lau, J. Y.

    1999-01-01

    Hepatitis C virus (HCV) nonstructural protein 3 (NS3) has been shown to possess protease and helicase activities and has also been demonstrated to spontaneously associate with nonstructural protein NS4A (NS4A) to form a stable complex. Previous attempts to produce the NS3/NS4A complex in recombinant baculovirus resulted in a protein complex that aggregated and precipitated in the absence of nonionic detergent and high salt. A single-chain form of the NS3/NS4A complex (His-NS4A21-32-GSGS-NS3-631) was constructed in which the NS4A core peptide is fused to the N-terminus of the NS3 protease domain as previously described (Taremi et al., 1998). This protein contains a histidine tagged NS4A peptide (a.a. 21-32) fused to the full-length NS3 (a.a. 3-631) through a flexible tetra amino acid linker. The recombinant protein was expressed to high levels in Escherichia coli, purified to homogeneity, and examined for NTPase, nucleic acid unwinding, and proteolytic activities. The single-chain recombinant NS3-NS4A protein possesses physiological properties equivalent to those of the NS3/NS4A complex except that this novel construct is stable, soluble and sixfold to sevenfold more active in unwinding duplex RNA. Comparison of the helicase activity of the single-chain recombinant NS3-NS4A with that of the full-length NS3 (without NS4A) and that of the helicase domain alone suggested that the presence of the protease domain and at least the NS4A core peptide are required for optimal unwinding activity. PMID:10386883

  5. On the chemistry of CS and NS in cometary comae

    NASA Astrophysics Data System (ADS)

    Canaves, M.; de Almeida, A.; Boice, D.; Sanzovo, G.

    The most fundamental scientific reason for studying comets is to retrieve information on their origin, relationship to interstellar and interplanetary material and implication for the formation of the Solar System or Cosmogony. The determination of the basic parameters of the nucleus and its activity and comp osition is desirable in order to establish a consistent database for comparative studies of comets and, as such, is vital for the safety and success of space missions. The objective of the present work is to contribute to the establishment of a unique description of the physical-chemical nature of the nucleus. We study carbon monosulfide (CS) - which is the only sulfur compound that persistently appears in cometary ultraviolet spectra and, therefore, seems to play a key role in sulfur photochemistry in cometary comae - and nitrogen monosulfide (NS) - the first cometary molecular species to contain both nitrogen and sulfur atoms which was recently observed by Irvine et al. (2000) in comet Hale -Bopp. The determination of the abundance of each such species helps to constrain the chemistry and physics of comets and hence their place and mode of origin of the nucleus. With this purpose in mind we have developed a multifluid chemical model of cometary comae (Boice 1990) with gas-phase chemical kinetics and gas dynamics to predict molecular abundance variations in a sensitive manner with cometocentric distance. We apply the model to the recent bright comets Hyakutake and Hale-Bopp at a heliocentric distance of 1 AU to study the abundances of CS and NS in their comae using a detailed photo and chemical reaction network with more than 100 species and over 1000 reactions. We conclude that the CS abundance in comets does not seem to vary much with the cometocentric distance. In particular, if NS is the daughter of an unknown long-lived parent molecular species, its production rate and abundance should be much larger than the obtained values. These results should be

  6. Evolution of laser-produced Sn extreme ultraviolet source diameter for high-brightness source

    SciTech Connect

    Roy, Amitava E-mail: aroy@barc.gov.in; Arai, Goki; Hara, Hiroyuki; Higashiguchi, Takeshi; Ohashi, Hayato; Sunahara, Atsushi; Li, Bowen; Dunne, Padraig; O'Sullivan, Gerry; Miura, Taisuke; Mocek, Tomas; Endo, Akira

    2014-08-18

    We have investigated the effect of irradiation of solid Sn targets with laser pulses of sub-ns duration and sub-mJ energy on the diameter of the extreme ultraviolet (EUV) emitting region and source conversion efficiency. It was found that an in-band EUV source diameter as low as 18 μm was produced due to the short scale length of a plasma produced by a sub-ns laser. Most of the EUV emission occurs in a narrow region with a plasma density close to the critical density value. Such EUV sources are suitable for high brightness and high repetition rate metrology applications.

  7. Non-structural protein NS3/NS3a is required for propagation of bluetongue virus in Culicoides sonorensis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Background: Bluetongue virus (BTV) causes non-contagious haemorrhagic disease in ruminants and is transmitted by Culicoides spp. biting midges. BTV encodes four non-structural proteins of which NS3/NS3a is functional in virus release. NS3/NS3a is not essential for in vitro virus replication. However...

  8. Progress on New Hepatitis C Virus Targets: NS2 and NS5A

    NASA Astrophysics Data System (ADS)

    Marcotrigiano, Joseph

    Hepatitis C virus (HCV) is a major global health problem, affecting about 170 million people worldwide. Chronic infection can lead to cirrhosis and liver cancer. The replication machine of HCV is a multi-subunit membrane associated complex, consisting of nonstructural proteins (NS2-5B), which replicate the viral RNA genome. The structures of NS5A and NS2 were recently determined. NS5A is an essential replicase component that also modulates numerous cellular processes ranging from innate immunity to cell growth and survival. The structure reveals a novel protein fold, a new zinc coordination motif, a disulfide bond and a dimer interface. Analysis of molecular surfaces suggests the location of the membrane interaction surface of NS5A, as well as hypothetical protein and RNA binding sites. NS2 is one of two virally encoded proteases that are required for processing the viral polyprotein into the mature nonstructural proteins. NS2 is a dimeric cysteine protease with two composite active sites. For each active site, the catalytic histidine and glutamate residues are contributed by one monomer and the nucleophilic cysteine by the other. The C-terminal residues remain coordinated in the two active sites, predicting an inactive post-cleavage form. The structure also reveals possible sites of membrane interaction, a rare cis-proline residue, and highly conserved dimer contacts. The novel features of both structures have changed the current view of HCV polyprotein replication and present new opportunities for antiviral drug design.

  9. The two-component NS2B-NS3 proteinase represses DNA unwinding activity of the West Nile virus NS3 helicase.

    PubMed

    Chernov, Andrei V; Shiryaev, Sergey A; Aleshin, Alexander E; Ratnikov, Boris I; Smith, Jeffrey W; Liddington, Robert C; Strongin, Alex Y

    2008-06-20

    Similar to many flavivirus types including Dengue and yellow fever viruses, the nonstructural NS3 multifunctional protein of West Nile virus (WNV) with an N-terminal serine proteinase domain and an RNA triphosphatase, an NTPase domain, and an RNA helicase in the C-terminal domain is implicated in both polyprotein processing and RNA replication and is therefore a promising drug target. To exhibit its proteolytic activity, NS3 proteinase requires the presence of the cofactor encoded by the upstream NS2B sequence. During our detailed investigation of the biology of the WNV helicase, we characterized the ATPase and RNA/DNA unwinding activities of the full-length NS2B-NS3 proteinase-helicase protein as well as the individual NS3 helicase domain lacking both the NS2B cofactor and the NS3 proteinase sequence and the individual NS3 proteinase-helicase lacking only the NS2B cofactor. We determined that both the NS3 helicase and NS3 proteinase-helicase constructs are capable of unwinding both the DNA and the RNA templates. In contrast, the full-length NS2B-NS3 proteinase-helicase unwinds only the RNA templates, whereas its DNA unwinding activity is severely repressed. Our data suggest that the productive, catalytically competent fold of the NS2B-NS3 proteinase moiety represents an essential component of the RNA-DNA substrate selectivity mechanism in WNV and, possibly, in other flaviviruses. Based on our data, we hypothesize that the mechanism we have identified plays a role yet to be determined in WNV replication occurring both within the virus-induced membrane-bound replication complexes in the host cytoplasm and in the nuclei of infected cells.

  10. In Situ Mosaic Brightness Correction

    NASA Technical Reports Server (NTRS)

    Deen, Robert G.; Lorre, Jean J.

    2012-01-01

    In situ missions typically have pointable, mast-mounted cameras, which are capable of taking panoramic mosaics comprised of many individual frames. These frames are mosaicked together. While the mosaic software applies radiometric correction to the images, in many cases brightness/contrast seams still exist between frames. This is largely due to errors in the radiometric correction, and the absence of correction for photometric effects in the mosaic processing chain. The software analyzes the overlaps between adjacent frames in the mosaic and determines correction factors for each image in an attempt to reduce or eliminate these brightness seams.

  11. High brightness picosecond electron gun

    SciTech Connect

    Merano, M.; Collin, S.; Renucci, P.; Gatri, M.; Sonderegger, S.; Crottini, A.; Ganiere, J.D.; Deveaud, B.

    2005-08-15

    We have developed a high brightness picosecond electron gun. We have used it to replace the thermionic electron gun of a commercial scanning electron microscope (SEM) in order to perform time-resolved cathodoluminescence experiments. Picosecond electron pulses are produced, at a repetition rate of 80.7 MHz, by femtosecond mode-locked laser pulses focused on a metal photocathode. This system has a normalized axial brightness of 93 A/cm{sup 2} sr kV, allowing for a spatial resolution of 50 nm in the secondary electron imaging mode of the SEM. The temporal width of the electron pulse is 12 ps.

  12. D-terms from generalized NS-NS fluxes in type II

    NASA Astrophysics Data System (ADS)

    Robbins, Daniel; Wrase, Timm

    2007-12-01

    Orientifolds of type II string theory admit a certain set of generalized NS-NS fluxes, including not only the three-form field strength H, but also metric and non-geometric fluxes, which are related to H by T-duality. We describe in general how these fluxes appear as parameters of an effective Script N = 1 supergravity theory in four dimensions, and in particular how certain generalized NS-NS fluxes can act as charges for R-R axions, leading to D-term contributions to the effective scalar potential. We illustrate these phenomena in type IIB with the example of a certain orientifold of T6/Bbb Z4.

  13. Physical Properties of Bright Comets

    NASA Astrophysics Data System (ADS)

    Pittichová, J.; Meech, K. J.

    2002-09-01

    We will show preliminary results from a program of long-term observation of the dust coma activity of bright comets. One and half years of observation of 32 selected comets in B, V, R, I filters are used for the study of the physical properties and the dust activity of their comae at a range of heliocentric distances from 0.99 to 8.61 AU. This enables us to compare the activity of different cometary nuclei at similar solar radiation conditions. As shown in the Table, the selected comets belong to different cometary populations from the point of view of their active age (near parabolic orbits versus short-period orbits, outbursts of brightness, disruption of nuclei) and orbital parameters (the eccentricity from 0.04 to 1.01 AU, the perihelion distance from 0.34 to 8.24 AU). The knowledge of physical properties of cometary nuclei and coma are very important to our understanding of the environment in the outer solar system during the era of formation. The comet dataset of 1128 images will enable us to study thermal evolution of the small dust particles, their dynamical parameters and size distribution as a function of time and grain size at different heliocentric distances. Our future goal is to model the near-nucleus particle region using a Finston-Probstein dust model. Since our observations are still in progress at this time we will present only preliminary results of brightness and color changes for several selected bright comets.

  14. Bright Transients discovered by PSST

    NASA Astrophysics Data System (ADS)

    Smith, K. W.; Wright, D.; Smartt, S. J.; Huber, M.; Chambers, K. C.; Flewelling, H.; Willman, M.; Primak, N.; Schultz, A.; Gibson, B.; Magnier, E.; Waters, C.; Tonry, J.; Wainscoat, R. J.; Denneau, L.; Stalder, B.; Heinze, A.; Sherstyuk, A.; Foley, R. J.; Jha, S. W.; Rest, A.; Scolnic, D.

    2016-04-01

    Seven bright transients have been discovered as part of the Pan-STARRS Survey for Transients (PSST). Information on all objects discovered by the Pan-STARRS Survey for Transients is available at http://star.pst.qub.ac.uk/ps1threepi/ (see Huber et al. ATel #7153).

  15. Bright Transients discovered by PSST

    NASA Astrophysics Data System (ADS)

    Smith, K. W.; Wright, D.; Smartt, S. J.; Huber, M.; Chambers, K. C.; Flewelling, H.; Willman, M.; Primak, N.; Schultz, A.; Gibson, B.; Magnier, E.; Waters, C.; Tonry, J.; Wainscoat, R. J.; Foley, R. J.; Jha, S. W.; Rest, A.; Scolnic, D.

    2016-03-01

    Seven bright transients have been discovered as part of the Pan-STARRS Survey for Transients (PSST). Information on all objects discovered by the Pan-STARRS Survey for Transients is available at http://star.pst.qub.ac.uk/ps1threepi/ (see Huber et al. ATel #7153).

  16. Bright Transients discovered by PSST

    NASA Astrophysics Data System (ADS)

    Smith, K. W.; Wright, D.; Smartt, S. J.; Young, D. R.; Huber, M.; Chambers, K. C.; Flewelling, H.; Willman, M.; Primak, N.; Schultz, A.; Gibson, B.; Magnier, E.; Waters, C.; Tonry, J.; Wainscoat, R. J.; Foley, R. J.; Jha, S. W.; Rest, A.; Scolnic, D.

    2016-08-01

    Six bright transients have been discovered as part of the Pan-STARRS Survey for Transients (PSST). Information on all objects discovered by the Pan-STARRS Survey for Transients is available at http://star.pst.qub.ac.uk/ps1threepi/ (see Huber et al. ATel #7153).

  17. Bright Beginnings. WWC Intervention Report

    ERIC Educational Resources Information Center

    What Works Clearinghouse, 2009

    2009-01-01

    Bright Beginnings is an early childhood curriculum, based in part on High/Scope[R] and Creative Curriculum[R], with an additional emphasis on literacy skills. The curriculum consists of nine thematic units designed to enhance children's cognitive, social, emotional, and physical development, and each unit includes concept maps, literacy lessons,…

  18. StarBright Learning Exchange

    ERIC Educational Resources Information Center

    Kalinowski, Michael

    2007-01-01

    This article features StarBright Learning Exchange, a program that provides a cross-cultural exchange between Australian and South African early childhood educators. The program was originated when its president, Carol Allen, and her colleague, Karen Williams, decided that they could no longer sit by and watch the unfolding social catastrophe that…

  19. Teradiode's high brightness semiconductor lasers

    NASA Astrophysics Data System (ADS)

    Huang, Robin K.; Chann, Bien; Burgess, James; Lochman, Bryan; Zhou, Wang; Cruz, Mike; Cook, Rob; Dugmore, Dan; Shattuck, Jeff; Tayebati, Parviz

    2016-03-01

    TeraDiode is manufacturing multi-kW-class ultra-high brightness fiber-coupled direct diode lasers for industrial applications. A fiber-coupled direct diode laser with a power level of 4,680 W from a 100 μm core diameter, <0.08 numerical aperture (NA) output fiber at a single center wavelength was demonstrated. Our TeraBlade industrial platform achieves world-record brightness levels for direct diode lasers. The fiber-coupled output corresponds to a Beam Parameter Product (BPP) of 3.5 mm-mrad and is the lowest BPP multi-kW-class direct diode laser yet reported. This laser is suitable for industrial materials processing applications, including sheet metal cutting and welding. This 4-kW fiber-coupled direct diode laser has comparable brightness to that of industrial fiber lasers and CO2 lasers, and is over 10x brighter than state-of-the-art direct diode lasers. We have also demonstrated novel high peak power lasers and high brightness Mid-Infrared Lasers.

  20. NetSim Project contributions to ns-3

    2012-05-01

    ns-3 is an external (non-LLNL) open-source framework for modeling computer networks. The LLNL NetSim project uses the ns-3 framework to address specific questions in computer network design, operation, and security. As part of the NetSim work, we develop bug fixes, deature enhancements, and new capabilities for the ns-3 framework. The virtual package referenced here, ns-3-contrib, consists of those developments we have (or will) contribute back to the ns-3 project in source code form, for inclusionmore » in future releases of ns-3.« less

  1. Brightness-equalized quantum dots

    PubMed Central

    Lim, Sung Jun; Zahid, Mohammad U.; Le, Phuong; Ma, Liang; Entenberg, David; Harney, Allison S.; Condeelis, John; Smith, Andrew M.

    2015-01-01

    As molecular labels for cells and tissues, fluorescent probes have shaped our understanding of biological structures and processes. However, their capacity for quantitative analysis is limited because photon emission rates from multicolour fluorophores are dissimilar, unstable and often unpredictable, which obscures correlations between measured fluorescence and molecular concentration. Here we introduce a new class of light-emitting quantum dots with tunable and equalized fluorescence brightness across a broad range of colours. The key feature is independent tunability of emission wavelength, extinction coefficient and quantum yield through distinct structural domains in the nanocrystal. Precise tuning eliminates a 100-fold red-to-green brightness mismatch of size-tuned quantum dots at the ensemble and single-particle levels, which substantially improves quantitative imaging accuracy in biological tissue. We anticipate that these materials engineering principles will vastly expand the optical engineering landscape of fluorescent probes, facilitate quantitative multicolour imaging in living tissue and improve colour tuning in light-emitting devices. PMID:26437175

  2. A New Sky Brightness Monitor

    NASA Astrophysics Data System (ADS)

    Crawford, David L.; McKenna, D.

    2006-12-01

    A good estimate of sky brightness and its variations throughout the night, the months, and even the years is an essential bit of knowledge both for good observing and especially as a tool in efforts to minimize sky brightness through local action. Hence a stable and accurate monitor can be a valuable and necessary tool. We have developed such a monitor, with the financial help of Vatican Observatory and Walker Management. The device is now undergoing its Beta test in preparation for production. It is simple, accurate, well calibrated, and automatic, sending its data directly to IDA over the internet via E-mail . Approximately 50 such monitors will be ready soon for deployment worldwide including most major observatories. Those interested in having one should enquire of IDA about details.

  3. Splicing of influenza A virus NS1 mRNA is independent of the viral NS1 protein.

    PubMed

    Robb, Nicole C; Jackson, David; Vreede, Frank T; Fodor, Ervin

    2010-09-01

    RNA segment 8 (NS) of influenza A virus encodes two proteins. The NS1 protein is translated from the unspliced primary mRNA transcript, whereas the second protein encoded by this segment, NS2/NEP, is translated from a spliced mRNA. Splicing of influenza NS1 mRNA is thought to be regulated so that the levels of NS2 spliced transcripts are approximately 10 % of total NS mRNA. Regulation of splicing of the NS1 mRNA has been studied at length, and a number of often-contradictory control mechanisms have been proposed. In this study, we used (32)P-labelled gene-specific primers to investigate influenza A NS1 mRNA splicing regulation. It was found that the efficiency of splicing of NS1 mRNA was maintained at similar levels in both virus infection and ribonucleoprotein-reconstitution assays, and NS2 mRNA comprised approximately 15 % of total NS mRNA in both assays. The effect of NS1 protein expression on the accumulation of viral NS2 mRNA and spliced cellular beta-globin mRNA was analysed, and it was found that NS1 protein expression reduced spliced beta-globin mRNA levels, but had no effect on the accumulation of NS2 mRNA. We conclude that the NS1 protein specifically inhibits the accumulation of cellular RNA polymerase II-driven mRNAs, but does not affect the splicing of its own viral NS1 mRNA.

  4. Iapetus Bright and Dark Terrains

    NASA Technical Reports Server (NTRS)

    1990-01-01

    Saturn's outermost large moon, Iapetus, has a bright, heavily cratered icy terrain and a dark terrain, as shown in this Voyager 2 image taken on August 22, 1981. Amazingly, the dark material covers precisely the side of Iapetus that leads in the direction of orbital motion around Saturn (except for the poles), whereas the bright material occurs on the trailing hemisphere and at the poles. The bright terrain is made of dirty ice, and the dark terrain is surfaced by carbonaceous molecules, according to measurements made with Earth-based telescopes. Iapetus' dark hemisphere has been likened to tar or asphalt and is so dark that no details within this terrain were visible to Voyager 2. The bright icy hemisphere, likened to dirty snow, shows many large impact craters. The closest approach by Voyager 2 to Iapetus was a relatively distant 600,000 miles, so that our best images, such as this, have a resolution of about 12 miles. The dark material is made of organic substances, probably including poisonous cyano compounds such as frozen hydrogen cyanide polymers. Though we know a little about the dark terrain's chemical nature, we do not understand its origin. Two theories have been developed, but neither is fully satisfactory--(1) the dark material may be organic dust knocked off the small neighboring satellite Phoebe and 'painted' onto the leading side of Iapetus as the dust spirals toward Saturn and Iapetus hurtles through the tenuous dust cloud, or (2) the dark material may be made of icy-cold carbonaceous 'cryovolcanic' lavas that were erupted from Iapetus' interior and then blackened by solar radiation, charged particles, and cosmic rays. A determination of the actual cause, as well as discovery of any other geologic features smaller than 12 miles across, awaits the Cassini Saturn orbiter to arrive in 2004.

  5. GPM Intercalibrated Radiometer Brightness Temperatures

    NASA Astrophysics Data System (ADS)

    Stocker, Erich Franz; Chou, Joyce

    2013-04-01

    One of the keys to consistent precipitation retrieval from passive microwave radiometer measurements (whether imagers or sounders) is accurate, long-term consistent brightness temperature retrievals. This becomes doubly important when there measurements are taken from radiometers on multiple platforms, from multiple agencies, with many different purposes. The Global Precipitation Measurement (GPM) mission addresses this issue directly with the production of intercalibrated brightness temperatures from all the partner satellites contributing to the GPM mission. These intercalibrated brightness temperatures are given the product designation: 1C within GPM. This paper will describe the GPM approach to intercalibration 1C products. The intercalibration and creation of the products uses a 5-step methodology: comparison of the partner standard products (either Tb or Ta) with the GPM reference standard; determination of adjustments that should be made to each product to create consistent brightness temperatures; re-orbitization of all orbits (in non-realtime) to be in the standard GPM south-south orbit; application of the adjustments to the partner provide 1B(or 1A) products; production of 1C products in HDF5 using a "standard" logical format for any radiometer regardless of its 1B format. This paper describes each of these steps and provides the background for them. It discusses in some detail the current 1C logical format and why this format facilitates use by downstream product algorithms and end-users. Most importantly it provides the analysis approach established by the GPM inter-calibration working group in establishing the adjustments to be made at the 1C level. Finally, using DMSP F16-18, it provides examples of the 1C products and discusses the adjustments that are made.

  6. LSST Site: Sky Brightness Data

    NASA Astrophysics Data System (ADS)

    Burke, Jamison; Claver, Charles

    2015-01-01

    The Large Synoptic Survey Telescope (LSST) is an upcoming robotic survey telescope. At the telescope site on Cerro Pachon in Chile there are currently three photodiodes and a Canon camera with a fisheye lens, and both the photodiodes and Canon monitor the night sky continuously. The NIST-calibrated photodiodes directly measure the flux from the sky, and the sky brightness can also be obtained from the Canon images via digital aperture photometry. Organizing and combining the two data sets gives nightly information of the development of sky brightness across a swath of the electromagnetic spectrum, from blue to near infrared light, and this is useful for accurately predicting the performance of the LSST. It also provides data for models of moonlight and twilight sky brightness. Code to accomplish this organization and combination was successfully written in Python, but due to the backlog of data not all of the nights were processed by the end of the summer.Burke was supported by the NOAO/KPNO Research Experiences for Undergraduates (REU) Program which is funded by the National Science Foundation Research Experiences for Undergraduates Program (AST-1262829).

  7. Purification and crystallization of dengue and West Nile virus NS2B–NS3 complexes

    SciTech Connect

    D’Arcy, Allan Chaillet, Maxime; Schiering, Nikolaus; Villard, Frederic; Lim, Siew Pheng; Lefeuvre, Peggy; Erbel, Paul

    2006-02-01

    Crystals of dengue serotype 2 and West Nile virus NS2B–NS3 protease complexes have been obtained and the crystals of both diffract to useful resolution. Sample homogeneity was essential for obtaining X-ray-quality crystals of the dengue protease. Controlled proteolysis produced a crystallizable fragment of the apo West Nile virus NS2B–NS3 and crystals were also obtained in the presence of a peptidic inhibitor. Both dengue and West Nile virus infections are an increasing risk to humans, not only in tropical and subtropical areas, but also in North America and parts of Europe. These viral infections are generally transmitted by mosquitoes, but may also be tick-borne. Infection usually results in mild flu-like symptoms, but can also cause encephalitis and fatalities. Approximately 2799 severe West Nile virus cases were reported this year in the United States, resulting in 102 fatalities. With this alarming increase in the number of West Nile virus infections in western countries and the fact that dengue virus already affects millions of people per year in tropical and subtropical climates, there is a real need for effective medicines. A possible therapeutic target to combat these viruses is the protease, which is essential for virus replication. In order to provide structural information to help to guide a lead identification and optimization program, crystallizations of the NS2B–NS3 protease complexes from both dengue and West Nile viruses have been initiated. Crystals that diffract to high resolution, suitable for three-dimensional structure determinations, have been obtained.

  8. A Macrocyclic Fluorophore Dimer with Flexible Linkers: Bright Excimer Emission with a Long Fluorescence Lifetime.

    PubMed

    Osaki, Hiroshi; Chou, Chih-Ming; Taki, Masayasu; Welke, Kai; Yokogawa, Daisuke; Irle, Stephan; Sato, Yoshikatsu; Higashiyama, Tetsuya; Saito, Shohei; Fukazawa, Aiko; Yamaguchi, Shigehiro

    2016-06-13

    Bright fluorescent molecules with long fluorescence lifetimes are important for the development of lifetime-based fluorescence imaging techniques. Herein, a molecular design is described for simultaneously attaining long fluorescence lifetime (τ) and high brightness (ΦF ×ɛ) in a system that features macrocyclic dimerization of fluorescent π-conjugated skeletons with flexible linkers. An alkylene-linked macrocyclic dimer of bis(thienylethynyl)anthracene was found to show excimer emission with a long fluorescence lifetime (τ≈19 ns) in solution, while maintaining high brightness. A comparison with various relevant derivatives revealed that the macrocyclic structure and the length of the alkylene chains play crucial roles in attaining these properties. In vitro time-gated imaging experiments were conducted as a proof-of-principle for the superiority of this macrocyclic fluorophore relative to the commercial fluorescent dye Alexa Fluor 488. PMID:27121201

  9. Brightness enhancement in a high-peak-power cladding-pumped Raman fiber amplifier.

    PubMed

    Sridharan, Arun Kumar; Heebner, John E; Messerly, Michael J; Dawson, Jay W; Beach, Raymond J; Barty, C P J

    2009-07-15

    We demonstrate a cladding-pumped Raman fiber amplifier (CPRFA) whose brightness-enhancement factor depends on the cladding-to-core diameter ratio. The pump and the signal are coupled independently into different input arms of a pump-signal combiner, and the output is spliced to the Raman amplifier fiber. The CPRFA generates 20 microJ, 7 ns pulses at 1100 nm at a 2.2 kHz repetition rate with 300 microJ (25.1 kW peak power) of input pump energy. The amplified signal's peak power is 2.77 kW, and the brightness-enhancement factor is 192--the highest peak power and brightness enhancement achieved in a CPRFA at any wavelength, to our knowledge.

  10. Brightness enhancement in a high-peak-power cladding-pumped Raman fiber amplifier.

    PubMed

    Sridharan, Arun Kumar; Heebner, John E; Messerly, Michael J; Dawson, Jay W; Beach, Raymond J; Barty, C P J

    2009-07-15

    We demonstrate a cladding-pumped Raman fiber amplifier (CPRFA) whose brightness-enhancement factor depends on the cladding-to-core diameter ratio. The pump and the signal are coupled independently into different input arms of a pump-signal combiner, and the output is spliced to the Raman amplifier fiber. The CPRFA generates 20 microJ, 7 ns pulses at 1100 nm at a 2.2 kHz repetition rate with 300 microJ (25.1 kW peak power) of input pump energy. The amplified signal's peak power is 2.77 kW, and the brightness-enhancement factor is 192--the highest peak power and brightness enhancement achieved in a CPRFA at any wavelength, to our knowledge. PMID:19823559

  11. Mechanistic and kinetic characterization of hepatitis C virus NS3 protein interactions with NS4A and protease inhibitors.

    PubMed

    Geitmann, Matthis; Dahl, Göran; Danielson, U Helena

    2011-01-01

    The mechanism and kinetics of the interactions between ligands and immobilized full-length hepatitis C virus (HCV) genotype 1a NS3 have been characterized by SPR biosensor technology. The NS3 interactions for a series of NS3 protease inhibitors as well as for the NS4A cofactor, represented by a peptide corresponding to the sequence interacting with the enzyme, were found to be heterogeneous. It may represent interactions with two stable conformations of the protein. The NS3-NS4A interaction consisted of a high-affinity (K(D) = 50 nM) and a low-affinity (K(D) = 2 µM) interaction, contributing equally to the overall binding. By immobilizing NS3 alone or together with NS4A it was shown that all inhibitors had a higher affinity for NS3 in the presence of NS4A. NS4A thus has a direct effect on the binding of inhibitors to NS3 and not only on catalysis. As predicted, the mechanism-based inhibitor VX 950 exhibited a time-dependent interaction with a slow formation of a stable complex. BILN 2061 or ITMN-191 showed no signs of time-dependent interactions, but ITMN-191 had the highest affinity of the tested compounds, with both the slowest dissociation (k(off)) and fastest association rate, closely followed by BILN 2061. The k(off) for the inhibitors correlated strongly with their NS3 protease inhibitory effect as well as with their effect on replication of viral proteins in replicon cell cultures, confirming the relevance of the kinetic data. This approach for obtaining kinetic and mechanistic data for NS3 protease inhibitor and cofactor interactions is expected to be of importance for understanding the characteristics of HCV NS3 functionality as well as for anti-HCV lead discovery and optimization. PMID:21194118

  12. High brightness beams and applications

    SciTech Connect

    Sheffield, R.L.

    1995-09-01

    This paper describes the present research on attaining intense bright electron beams. Thermionic systems are briefly covered. Recent and past results from the photoinjector programs are given. The performance advantages and difficulties presently faced by researchers using photoinjectors is discussed. The progress that has been made in photocathode materials, both in lifetime and quantum efficiency, is covered. Finally, a discussion of emittance measurements of photoinjector systems and how the measurement is complicated by the non-thermal nature of the electron beam is presented.

  13. Determinants of Dengue Virus NS4A Protein Oligomerization

    PubMed Central

    Lee, Chia Min; Xie, Xuping; Zou, Jing; Li, Shi-Hua; Lee, Michelle Yue Qi; Dong, Hongping; Qin, Cheng-Feng; Kang, Congbao

    2015-01-01

    ABSTRACT Flavivirus NS4A protein induces host membrane rearrangement and functions as a replication complex component. The molecular details of how flavivirus NS4A exerts these functions remain elusive. Here, we used dengue virus (DENV) as a model to characterize and demonstrate the biological relevance of flavivirus NS4A oligomerization. DENV type 2 (DENV-2) NS4A protein forms oligomers in infected cells or when expressed alone. Deletion mutagenesis mapped amino acids 50 to 76 (spanning the first transmembrane domain [TMD1]) of NS4A as the major determinant for oligomerization, while the N-terminal 50 residues contribute only slightly to the oligomerization. Nuclear magnetic resonance (NMR) analysis of NS4A amino acids 17 to 80 suggests that residues L31, L52, E53, G66, and G67 could participate in oligomerization. Ala substitution for 15 flavivirus conserved NS4A residues revealed that these amino acids are important for viral replication. Among the 15 mutated NS4A residues, 2 amino acids (E50A and G67A) are located within TMD1. Both E50A and G67A attenuated viral replication, decreased NS4A oligomerization, and reduced NS4A protein stability. In contrast, NS4A oligomerization was not affected by the replication-defective mutations (R12A, P49A, and K80A) located outside TMD1. trans complementation experiments showed that expression of wild-type NS4A alone was not sufficient to rescue the replication-lethal NS4A mutants. However, the presence of DENV-2 replicons could partially restore the replication defect of some lethal NS4A mutants (L26A and K80A), but not others (L60A and E122A), suggesting an unidentified mechanism governing the outcome of complementation in a mutant-dependent manner. Collectively, the results have demonstrated the importance of TMD1-mediated NS4A oligomerization in flavivirus replication. IMPORTANCE We report that DENV NS4A forms oligomers. Such NS4A oligomerization is mediated mainly through amino acids 50 to 76 (spanning the first

  14. Dengue virus NS1 triggers endothelial permeability and vascular leak that is prevented by NS1 vaccination.

    PubMed

    Beatty, P Robert; Puerta-Guardo, Henry; Killingbeck, Sarah S; Glasner, Dustin R; Hopkins, Kaycie; Harris, Eva

    2015-09-01

    The four dengue virus serotypes (DENV1 to DENV4) are mosquito-borne flaviviruses that cause up to ~100 million cases of dengue annually worldwide. Severe disease is thought to result from immunopathogenic processes involving serotype cross-reactive antibodies and T cells that together induce vasoactive cytokines, causing vascular leakage that leads to shock. However, no viral proteins have been directly implicated in triggering endothelial permeability, which results in vascular leakage. DENV nonstructural protein 1 (NS1) is secreted and circulates in patients' blood during acute infection; high levels of NS1 are associated with severe disease. We show that inoculation of mice with DENV NS1 alone induces both vascular leakage and production of key inflammatory cytokines. Furthermore, simultaneous administration of NS1 with a sublethal dose of DENV2 results in a lethal vascular leak syndrome. We also demonstrate that NS1 from DENV1, DENV2, DENV3, and DENV4 triggers endothelial barrier dysfunction, causing increased permeability of human endothelial cell monolayers in vitro. These pathogenic effects of physiologically relevant amounts of NS1 in vivo and in vitro were blocked by NS1-immune polyclonal mouse serum or monoclonal antibodies to NS1, and immunization of mice with NS1 from DENV1 to DENV4 protected against lethal DENV2 challenge. These findings add an important and previously overlooked component to the causes of dengue vascular leak, identify a new potential target for dengue therapeutics, and support inclusion of NS1 in dengue vaccines. PMID:26355030

  15. NMR analysis of a novel enzymatically active unlinked dengue NS2B-NS3 protease complex.

    PubMed

    Kim, Young Mee; Gayen, Shovanlal; Kang, CongBao; Joy, Joma; Huang, Qiwei; Chen, Angela Shuyi; Wee, John Liang Kuan; Ang, Melgious Jin Yan; Lim, Huichang Annie; Hung, Alvin W; Li, Rong; Noble, Christian G; Lee, Le Tian; Yip, Andy; Wang, Qing-Yin; Chia, Cheng San Brian; Hill, Jeffrey; Shi, Pei-Yong; Keller, Thomas H

    2013-05-01

    The dengue virus (DENV) is a mosquito-borne pathogen responsible for an estimated 100 million human infections annually. The viral genome encodes a two-component trypsin-like protease that contains the cofactor region from the nonstructural protein NS2B and the protease domain from NS3 (NS3pro). The NS2B-NS3pro complex plays a crucial role in viral maturation and has been identified as a potential drug target. Using a DENV protease construct containing NS2B covalently linked to NS3pro via a Gly4-Ser-Gly4 linker ("linked protease"), previous x-ray crystal structures show that the C-terminal fragment of NS2B is remote from NS3pro and exists in an open state in the absence of an inhibitor; however, in the presence of an inhibitor, NS2B complexes with NS3pro to form a closed state. This linked enzyme produced NMR spectra with severe signal overlap and line broadening. To obtain a protease construct with a resolved NMR spectrum, we expressed and purified an unlinked protease complex containing a 50-residue segment of the NS2B cofactor region and NS3pro without the glycine linker using a coexpression system. This unlinked protease complex was catalytically active at neutral pH in the absence of glycerol and produced dispersed cross-peaks in a (1)H-(15)N heteronuclear single quantum correlation spectrum that enabled us to conduct backbone assignments using conventional techniques. In addition, titration with an active-site peptide aldehyde inhibitor and paramagnetic relaxation enhancement studies demonstrated that the unlinked DENV protease exists predominantly in a closed conformation in solution. This protease complex can serve as a useful tool for drug discovery against DENV.

  16. How Bright Can Supernovae Get?

    NASA Astrophysics Data System (ADS)

    Kohler, Susanna

    2016-04-01

    Supernovae enormous explosions associated with the end of a stars life come in a variety of types with different origins. A new study has examined how the brightest supernovae in the Universe are produced, and what limits might be set on their brightness.Ultra-Luminous ObservationsRecent observations have revealed many ultra-luminous supernovae, which haveenergies that challenge our abilities to explain them usingcurrent supernova models. An especially extreme example is the 2015 discovery of the supernova ASASSN-15lh, which shone with a peak luminosity of ~2*1045 erg/s, nearly a trillion times brighter than the Sun. ASASSN-15lh radiated a whopping ~2*1052 erg in the first four months after its detection.How could a supernova that bright be produced? To explore the answer to that question, Tuguldur Sukhbold and Stan Woosley at University of California, Santa Cruz, have examined the different sources that could produce supernovae and calculated upper limits on the potential luminosities ofeach of these supernova varieties.Explosive ModelsSukhbold and Woosley explore multiple different models for core-collapse supernova explosions, including:Prompt explosionA stars core collapses and immediately explodes.Pair instabilityElectron/positron pair production at a massive stars center leads to core collapse. For high masses, radioactivity can contribute to delayed energy output.Colliding shellsPreviously expelled shells of material around a star collide after the initial explosion, providing additional energy release.MagnetarThe collapsing star forms a magnetar a rapidly rotating neutron star with an incredibly strong magnetic field at its core, which then dumps energy into the supernova ejecta, further brightening the explosion.They then apply these models to different types of stars.Setting the LimitThe authors show that the light curve of ASASSN-15lh (plotted in orange) can be described by a model (black curve) in which a magnetar with an initial spin period of 0.7 ms

  17. [Bright light therapy for elderly].

    PubMed

    Okawa, Masako

    2015-06-01

    Bright light therapy (BLT) holds considerable promise for sleep problems in the elderly. BLT for community-dwelling patients with Alzheimer's disease showed significant improvement in sleep parameters. In the institutional setting, BLT was effective in reducing daytime nap duration. Morning BLT was found to advance the peak circadian rhythm and increase activity level in daytime and melatonin level at night. Light therapy could be used in combination with other nonpharmacological methods such as social activities, outside walking, physical exercises, which showed greater effects than independent BLT on sleep and cognitive function. BLT treatment strategy was proposed in the present paper. We should pay more attentions to BLT in community setting for mental and physical well-being. PMID:26065132

  18. Strong pollinator-mediated selection for increased flower brightness and contrast in a deceptive orchid.

    PubMed

    Sletvold, Nina; Trunschke, Judith; Smit, Mart; Verbeek, Jeffrey; Ågren, Jon

    2016-03-01

    Contrasting flower color patterns that putatively attract or direct pollinators toward a reward are common among angiosperms. In the deceptive orchid Anacamptis morio, the lower petal, which makes up most of the floral display, has a light central patch with dark markings. Within populations, there is pronounced variation in petal brightness, patch size, amount of dark markings, and contrast between patch and petal margin. We tested whether pollinators mediate selection on these color traits and on morphology (plant height, number of flowers, corolla size, spur length), and whether selection is consistent with facilitated or negative frequency-dependent pollination. Pollinators mediated strong selection for increased petal brightness (Δβpoll = 0.42) and contrast (Δβpoll = 0.51). Pollinators also tended to mediate stabilizing selection on brightness (Δγpoll = -0.27, n.s.) favoring the most common phenotype in the population. Selection for reduced petal brightness among hand-pollinated plants indicated a fitness cost associated with brightness. The results demonstrate that flower color traits influence pollination success and seed production in A. morio, indicating that they affect attractiveness to pollinators, efficiency of pollen transfer, or both. The documented selection is consistent with facilitated pollination and selection for color convergence toward cooccurring rewarding species. PMID:26878831

  19. Strong pollinator-mediated selection for increased flower brightness and contrast in a deceptive orchid.

    PubMed

    Sletvold, Nina; Trunschke, Judith; Smit, Mart; Verbeek, Jeffrey; Ågren, Jon

    2016-03-01

    Contrasting flower color patterns that putatively attract or direct pollinators toward a reward are common among angiosperms. In the deceptive orchid Anacamptis morio, the lower petal, which makes up most of the floral display, has a light central patch with dark markings. Within populations, there is pronounced variation in petal brightness, patch size, amount of dark markings, and contrast between patch and petal margin. We tested whether pollinators mediate selection on these color traits and on morphology (plant height, number of flowers, corolla size, spur length), and whether selection is consistent with facilitated or negative frequency-dependent pollination. Pollinators mediated strong selection for increased petal brightness (Δβpoll = 0.42) and contrast (Δβpoll = 0.51). Pollinators also tended to mediate stabilizing selection on brightness (Δγpoll = -0.27, n.s.) favoring the most common phenotype in the population. Selection for reduced petal brightness among hand-pollinated plants indicated a fitness cost associated with brightness. The results demonstrate that flower color traits influence pollination success and seed production in A. morio, indicating that they affect attractiveness to pollinators, efficiency of pollen transfer, or both. The documented selection is consistent with facilitated pollination and selection for color convergence toward cooccurring rewarding species.

  20. A Closer Look at the NS1 of Influenza Virus

    PubMed Central

    Dundon, William G.; Capua, Ilaria

    2009-01-01

    The Non-Structural 1 (NS1) protein is a multifactorial protein of type A influenza viruses that plays an important role in the virulence of the virus. A large amount of what we know about this protein has been obtained from studies using human influenza isolates and, consequently, the human NS1 protein. The current global interest in avian influenza, however, has highlighted a number of sequence and functional differences between the human and avian NS1. This review discusses these differences in addition to describing potential uses of NS1 in the management and control of avian influenza outbreaks. PMID:21994582

  1. Anthracene-based inhibitors of dengue virus NS2B-NS3 protease.

    PubMed

    Tomlinson, Suzanne M; Watowich, Stanley J

    2011-02-01

    Dengue virus (DENV) is a mosquito-borne flavivirus that has strained global healthcare systems throughout tropical and subtropical regions of the world. In addition to plaguing developing nations, it has re-emerged in several developed countries with recent outbreaks in the USA (CDC, 2010), Australia (Hanna et al., 2009), Taiwan (Kuan et al., 2010) and France (La Ruche et al., 2010). DENV infection can cause significant disease, including dengue fever, dengue hemorrhagic fever, dengue shock syndrome, and death. There are no approved vaccines or antiviral therapies to prevent or treat dengue-related illnesses. However, the viral NS2B-NS3 protease complex provides a strategic target for antiviral drug development since NS3 protease activity is required for virus replication. Recently, we reported two compounds with inhibitory activity against the DENV protease in vitro and antiviral activity against dengue 2 (DEN2V) in cell culture (Tomlinson et al., 2009a). Analogs of one of the lead compounds were purchased, tested in protease inhibition assays, and the data evaluated with detailed kinetic analyses. A structure activity relationship (SAR) identified key atomic determinants (i.e. functional groups) important for inhibitory activity. Four "second series" analogs were selected and tested to validate our SAR and structural models. Here, we report improvements to inhibitory activity ranging between ∼2- and 60-fold, resulting in selective low micromolar dengue protease inhibitors.

  2. Quantum communication with macroscopically bright nonclassical states.

    PubMed

    Usenko, Vladyslav C; Ruppert, Laszlo; Filip, Radim

    2015-11-30

    We analyze homodyne detection of macroscopically bright multimode nonclassical states of light and propose their application in quantum communication. We observe that the homodyne detection is sensitive to a mode-matching of the bright light to the highly intense local oscillator. Unmatched bright modes of light result in additional noise which technically limits detection of Gaussian entanglement at macroscopic level. When the mode-matching is sufficient, we show that multimode quantum key distribution with bright beams is feasible. It finally merges the quantum communication with classical optical technology of visible beams of light.

  3. Balance of RNA sequence requirement and NS3/NS3a expression of segment 10 of orbiviruses.

    PubMed

    Feenstra, Femke; van Gennip, René G P; Schreuder, Myrte; van Rijn, Piet A

    2016-02-01

    Orbiviruses are insect-transmitted, non-enveloped viruses with a ten-segmented dsRNA genome of which the bluetongue virus (BTV) is the prototype. Viral non-structural protein NS3/NS3a is encoded by genome segment 10 (Seg-10), and is involved in different virus release mechanisms. This protein induces specific release via membrane disruptions and budding in both insect and mammalian cells, but also the cytopathogenic release that is only seen in mammalian cells. NS3/NS3a is not essential for virus replication in vitro with BTV Seg-10 containing RNA elements essential for virus replication, even if protein is not expressed. Recently, new BTV serotypes with distinct NS3/NS3a sequence and cell tropism have been identified. Multiple studies have hinted at the importance of Seg-10 in orbivirus replication, but the exact prerequisites are still unknown. Here, more insight is obtained with regard to the needs for orbivirus Seg-10 and the balance between protein expression and RNA elements. Multiple silent mutations in the BTV NS3a ORF destabilized Seg-10, resulting in deletions and sequences originating from other viral segments being inserted, indicating strong selection at the level of RNA during replication in mammalian cells in vitro. The NS3a ORFs of other orbiviruses were successfully exchanged in BTV1 Seg-10, resulting in viable chimeric viruses. NS3/NS3a proteins in these chimeric viruses were generally functional in mammalian cells, but not in insect cells. NS3/NS3a of the novel BTV serotypes 25 and 26 affected virus release from Culicoides cells, which might be one of the reasons for their distinct cell tropism. PMID:26644214

  4. A ns-Pulse Laser Microthruster

    SciTech Connect

    Phipps, Claude R.; Luke, James R.

    2006-05-02

    We have developed a prototype device which demonstrates the feasibility of using ns-duration laser pulses in a laser microthruster. Relative to the ms-duration thrusters which we have demonstrated in the past, this change offers the use of any target material, the use of reflection-mode target illumination, and adjustable specific impulse. Specific impulse is adjusted by varying laser intensity on target. In this way, we were able to vary specific impulse from 200s to 3,200s on gold. We used a Concepts Research, Inc. microchip laser with 170mW average optical power, 8kHz repetition rate and 20{mu}J pulse energy for many of the measurements. Thrust was in the 100nN - 1{mu}N range for all the work, requiring development of an extremely sensitive, low-noise thrust stand. We will discuss the design of metallic fuel delivery systems. Ablation efficiency near 100% was observed. Results obtained on metallic fuel systems agreed with simulations. We also report time-of-flight measurements on ejected metal ions, which gave velocities up to 80km/s.

  5. Brazilian Flavivirus phylogeny based on NS5.

    PubMed

    Baleotti, Flúvia Graciela; Moreli, Marcos Lázaro; Figueiredo, Luiz Tadeu Moraes

    2003-04-01

    In this work, a comprehensive phylogenetic study based on 600 base pair nucleotide and on putative 200 amino acid sequences of NS5 was carried out in order to establish genetic relationships among 15 strains of 10 Brazilian flaviviruses: Bussuquara, Cacipacore, dengue type 1, 2 and 4, Iguape, Ilheus, Rocio, Saint Louis encephalitis (SLE), and yellow fever. Phylogenetic trees were created by neighbor-joining and maximum parsimony methods. These trees showed Brazilian flaviviruses grouped into three main branches: yellow fever branch, dengue branch subdivided in types 1, 2 and 4 branches, and Japanese encephalitis virus (JEV) complex branch including SLE virus strains, Cacipacore, Iguape, Rocio, Ilheus and Bussuquara. Viruses transmitted by Aedes mosquitoes, such as dengue and urban yellow fever, that are also the only Flavivirus causing hemorrhagic fevers in Brazil, were grouped in the same cluster. Encephalitis associated viruses, transmitted by Culex mosquitoes such as JEV complex branch including SLE virus strains, Cacipacore, Iguape, Rocio, Ilheus and Bussuquara were also grouped in the same clade.

  6. Synergistic Activity of Combined NS5A Inhibitors

    PubMed Central

    Nower, Peter T.; Gao, Min; Fridell, Robert; Wang, Chunfu; Hewawasam, Piyasena; Lopez, Omar; Tu, Yong; Meanwell, Nicholas A.; Belema, Makonen; Roberts, Susan B.; Cockett, Mark; Sun, Jin-Hua

    2015-01-01

    Daclatasvir (DCV) is a first-in-class hepatitis C virus (HCV) nonstructural 5A replication complex inhibitor (NS5A RCI) that is clinically effective in interferon-free combinations with direct-acting antivirals (DAAs) targeting alternate HCV proteins. Recently, we reported NS5A RCI combinations that enhance HCV inhibitory potential in vitro, defining a new class of HCV inhibitors termed NS5A synergists (J. Sun, D. R. O’Boyle II, R. A. Fridell, D. R. Langley, C. Wang, S. Roberts, P. Nower, B. M. Johnson F. Moulin, M. J. Nophsker, Y. Wang, M. Liu, K. Rigat, Y. Tu, P. Hewawasam, J. Kadow, N. A. Meanwell, M. Cockett, J. A. Lemm, M. Kramer, M. Belema, and M. Gao, Nature 527:245–248, 2015, doi:10.1038/nature15711). To extend the characterization of NS5A synergists, we tested new combinations of DCV and NS5A synergists against genotype (gt) 1 to 6 replicons and gt 1a, 2a, and 3a viruses. The kinetics of inhibition in HCV-infected cells treated with DCV, an NS5A synergist (NS5A-Syn), or a combination of DCV and NS5A-Syn were distinctive. Similar to activity observed clinically, DCV caused a multilog drop in HCV, followed by rebound due to the emergence of resistance. DCV–NS5A-Syn combinations were highly efficient at clearing cells of viruses, in line with the trend seen in replicon studies. The retreatment of resistant viruses that emerged using DCV monotherapy with DCV–NS5A-Syn resulted in a multilog drop and rebound in HCV similar to the initial decline and rebound observed with DCV alone on wild-type (WT) virus. A triple combination of DCV, NS5A-Syn, and a DAA targeting the NS3 or NS5B protein cleared the cells of viruses that are highly resistant to DCV. Our data support the observation that the cooperative interaction of DCV and NS5A-Syn potentiates both the genotype coverage and resistance barrier of DCV, offering an additional DAA option for combination therapy and tools for explorations of NS5A function. PMID:26711745

  7. The dusty side of planetary nebulae: a HerPlaNS view

    NASA Astrophysics Data System (ADS)

    Ueta, Toshiya; Ladjal, Djazia; pre=", HerPlaNS Team

    2016-07-01

    HerPlaNS (Herschel Planetary Nebula Survey) is a far-IR imaging/spectroscopic survey of planetary nebulae (PNe) using the Herschel Space Observatory. In this presentation, we review our investigation into the physical properties of the cold dust component of the target PNe. We find that the far-IR surface brightness emission from PNe is generally dominated by thermal dust emission, which exhibits particular characteristics in terms of the dust emissivity and dust temperature compared with dust grains found elsewhere. The PN dust displays little variation in the emissivity while a large spread in the temperature, suggesting the presence of rather homogeneous dust chemistry and size distribution in the circumstellar environs.

  8. Optimising the efficiency of pulsed diode pumped Yb:YAG laser amplifiers for ns pulse generation.

    PubMed

    Ertel, K; Banerjee, S; Mason, P D; Phillips, P J; Siebold, M; Hernandez-Gomez, C; Collier, J C

    2011-12-19

    We present a numerical model of a pulsed, diode-pumped Yb:YAG laser amplifier for the generation of high energy ns-pulses. This model is used to explore how optical-to-optical efficiency depends on factors such as pump duration, pump spectrum, pump intensity, doping concentration, and operating temperature. We put special emphasis on finding ways to achieve high efficiency within the practical limitations imposed by real-world laser systems, such as limited pump brightness and limited damage fluence. We show that a particularly advantageous way of improving efficiency within those constraints is operation at cryogenic temperature. Based on the numerical findings we present a concept for a scalable amplifier based on an end-pumped, cryogenic, gas-cooled multi-slab architecture.

  9. Spatial Brightness Perception of Trichromatic Stimuli

    SciTech Connect

    Royer, Michael P.; Houser, Kevin W.

    2012-11-16

    An experiment was conducted to examine the effect of tuning optical radiation on brightness perception for younger (18-25 years of age) and older (50 years of age or older) observers. Participants made forced-choice evaluations of the brightness of a full factorial of stimulus pairs selected from two groups of four metameric stimuli. The large-field stimuli were created by systematically varying either the red or the blue primary of an RGB LED mixture. The results indicate that light stimuli of equal illuminance and chromaticity do not appear equally bright to either younger or older subjects. The rank-order of brightness is not predicted by any current model of human vision or theory of brightness perception including Scotopic to Photopic or Cirtopic to Photopic ratio theory, prime color theory, correlated color temperature, V(λ)-based photometry, color quality metrics, linear brightness models, or color appearance models. Age may affect brightness perception when short-wavelength primaries are used, especially those with a peak wavelength shorter than 450 nm. The results suggest further development of metrics to predict brightness perception is warranted, and that including age as a variable in predictive models may be valuable.

  10. Contrast adaptation to luminance and brightness modulations.

    PubMed

    Nagai, Takehiro; Nakayama, Kazuki; Kawashima, Yuki; Yamauchi, Yasuki

    2016-03-01

    Perceptual brightness and color contrast decrease after seeing a light temporally modulating along a certain direction in a color space, a phenomenon known as contrast adaptation. We investigated whether contrast adaptation along the luminance direction arises from modulation of luminance signals or apparent brightness signals. The stimulus consisted of two circles on a gray background presented on a CRT monitor. In the adaptation phase, the luminance and chromaticity of one circle were temporally modulated, while the other circle was kept at a constant luminance and color metameric with an equal-energy white. We employed two types of temporal modulations, namely, in luminance and brightness. Chromaticity was sinusoidally modulated along the L-M axis, leading to dissociation between luminance and brightness (the Helmholtz-Kohlrausch effect). In addition, luminance modulation was minimized in the brightness modulation, while brightness modulation was minimized in the luminance modulation. In the test phase, an asymmetric matching method was used to measure the magnitude of contrast adaptation for both modulations. Our results showed that, although contrast adaptation along the luminance direction occurred for both modulations, contrast adaptation for luminance modulation was significantly stronger than that for the brightness modulation regardless of the temporal frequency of the adaptation modulation. These results suggest that luminance modulation is more influential in contrast adaptation than brightness modulation.

  11. Hepatitis C Virus NS3 Inhibitors: Current and Future Perspectives

    PubMed Central

    Akimitsu, Nobuyoshi

    2013-01-01

    Currently, hepatitis C virus (HCV) infection is considered a serious health-care problem all over the world. A good number of direct-acting antivirals (DAAs) against HCV infection are in clinical progress including NS3-4A protease inhibitors, RNA-dependent RNA polymerase inhibitors, and NS5A inhibitors as well as host targeted inhibitors. Two NS3-4A protease inhibitors (telaprevir and boceprevir) have been recently approved for the treatment of hepatitis C in combination with standard of care (pegylated interferon plus ribavirin). The new therapy has significantly improved sustained virologic response (SVR); however, the adverse effects associated with this therapy are still the main concern. In addition to the emergence of viral resistance, other targets must be continually developed. One such underdeveloped target is the helicase portion of the HCV NS3 protein. This review article summarizes our current understanding of HCV treatment, particularly with those of NS3 inhibitors. PMID:24282816

  12. Ultrastructure of Kunjin virus-infected cells: colocalization of NS1 and NS3 with double-stranded RNA, and of NS2B with NS3, in virus-induced membrane structures.

    PubMed Central

    Westaway, E G; Mackenzie, J M; Kenney, M T; Jones, M K; Khromykh, A A

    1997-01-01

    The subcellular location of the nonstructural proteins NS1, NS2B, and NS3 in Vero cells infected with the flavivirus Kunjin was investigated using indirect immunofluorescence and cryoimmunoelectron microscopy with monospecific antibodies. Comparisons were also made by dual immunolabelling using antibodies to double-stranded RNA (dsRNA), the putative template in the flavivirus replication complex. At 8 h postinfection, the immunofluorescent patterns showed NS1, NS2B, NS3, and dsRNA located in a perinuclear rim with extensions into the peripheral cytoplasm. By 16 h, at the end of the latent period, all patterns had changed to some discrete perinuclear foci associated with a thick cytoplasmic reticulum. By 24 h, this localization in perinuclear foci was more apparent and some foci were dual labelled with antibodies to dsRNA. In immuno-gold-labelled cryosections of infected cells at 24 h, all antibodies were associated with clusters of induced membrane structures in the perinuclear region. Two important and novel observations were made. First, one set of induced membranes comprised vesicle packets of smooth membranes dual labelled with anti-dsRNA and anti-NS1 or anti-NS3 antibodies. Second, adjacent masses of paracrystalline arrays or of convoluted smooth membranes, which appeared to be structurally related, were strongly labelled only with anti-NS2B and anti-NS3 antibodies. Paired membranes similar in appearance to the rough endoplasmic reticulum were also labelled, but less strongly, with antibodies to the three nonstructural proteins. Other paired membranes adjacent to the structures discussed above enclosed accumulated virus particles but were not labelled with any of the four antibodies. The collection of induced membranes may represent virus factories in which translation, RNA synthesis, and virus assembly occur. PMID:9261387

  13. Flavonoids as noncompetitive inhibitors of Dengue virus NS2B-NS3 protease: inhibition kinetics and docking studies.

    PubMed

    de Sousa, Lorena Ramos Freitas; Wu, Hongmei; Nebo, Liliane; Fernandes, João Batista; da Silva, Maria Fátima das Graças Fernandes; Kiefer, Werner; Kanitz, Manuel; Bodem, Jochen; Diederich, Wibke E; Schirmeister, Tanja; Vieira, Paulo Cezar

    2015-02-01

    NS2B-NS3 is a serine protease of the Dengue virus considered a key target in the search for new antiviral drugs. In this study flavonoids were found to be inhibitors of NS2B-NS3 proteases of the Dengue virus serotypes 2 and 3 with IC50 values ranging from 15 to 44 μM. Agathisflavone (1) and myricetin (4) turned out to be noncompetitive inhibitors of dengue virus serotype 2 NS2B-NS3 protease with Ki values of 11 and 4.7 μM, respectively. Docking studies propose a binding mode of the flavonoids in a specific allosteric binding site of the enzyme. Analysis of biomolecular interactions of quercetin (5) with NT647-NHS-labeled Dengue virus serotype 3 NS2B-NS3 protease by microscale thermophoresis experiments, yielded a dissociation constant KD of 20 μM. Our results help to understand the mechanism of inhibition of the Dengue virus serine protease by flavonoids, which is essential for the development of improved inhibitors.

  14. Bright Sparks of Our Future!

    NASA Astrophysics Data System (ADS)

    Riordan, Naoimh

    2016-04-01

    My name is Naoimh Riordan and I am the Vice Principal of Rockboro Primary School in Cork City, South of Ireland. I am a full time class primary teacher and I teach 4th class, my students are aged between 9-10 years. My passion for education has developed over the years and grown towards STEM (Science, Technology, Engineering and Mathematics) subjects. I believe these subjects are the way forward for our future. My passion and beliefs are driven by the unique after school programme that I have developed. It is titled "Sparks" coming from the term Bright Sparks. "Sparks" is an after school programme with a difference where the STEM subjects are concentrated on through lessons such as Science, Veterinary Science Computer Animation /Coding, Eco engineering, Robotics, Magical Maths, Chess and Creative Writing. All these subjects are taught through activity based learning and are one-hour long each week for a ten-week term. "Sparks" is fully inclusive and non-selective which gives all students of any level of ability an opportunity to engage into these subjects. "Sparks" is open to all primary students in County Cork. The "Sparks" after school programme is taught by tutors from the different Universities and Colleges in Cork City. It works very well because the tutor brings their knowledge, skills and specialised equipment from their respective universities and in turn the tutor gains invaluable teaching practise, can trial a pilot programme in a chosen STEM subject and gain an insight into what works in the physical classroom.

  15. High-brightness, high-current density cathode for induction linac FELs

    SciTech Connect

    Turner, W.C.; Chen, Y.-J.; Nexsen, W.E.; Green, M.C.; Miram, G.; Nordquist, A.V.

    1988-09-28

    We have recently initiated an investigation to determine the intrinsic operating limits of an osmium coated dispenser cathode for use in free-electron lasers (FELs) driven by an induction linear accelerator. The experimental apparatus consists of a 5.1-cm-diam osmium coated dispenser cathode driven by a 250-kV, 10-..cap omega.., 35-ns Blumlein pulse line. The pepper pot technique is used to measure intrinsic cathode brightness and uniformity. Recent measurements have yielded brightness values exceeding 1 /times/ 10/sup 10/ A/m/sup 2/rad/sup 2/ for current densities up to 140 A/cm/sup 2/. We have also obtained quantitative data on cathode poisoning caused by a number of chemical agents of interest in the induction linac environment. 7 refs., 8 figs., 2 tabs.

  16. Novel Dengue Virus NS2B/NS3 Protease Inhibitors

    PubMed Central

    Wu, Hongmei; Bock, Stefanie; Snitko, Mariya; Berger, Thilo; Weidner, Thomas; Holloway, Steven; Kanitz, Manuel; Diederich, Wibke E.; Steuber, Holger; Walter, Christof; Hofmann, Daniela; Weißbrich, Benedikt; Spannaus, Ralf; Acosta, Eliana G.; Bartenschlager, Ralf; Engels, Bernd; Schirmeister, Tanja

    2014-01-01

    Dengue fever is a severe, widespread, and neglected disease with more than 2 million diagnosed infections per year. The dengue virus NS2B/NS3 protease (PR) represents a prime target for rational drug design. At the moment, there are no clinical PR inhibitors (PIs) available. We have identified diaryl (thio)ethers as candidates for a novel class of PIs. Here, we report the selective and noncompetitive inhibition of the serotype 2 and 3 dengue virus PR in vitro and in cells by benzothiazole derivatives exhibiting 50% inhibitory concentrations (IC50s) in the low-micromolar range. Inhibition of replication of DENV serotypes 1 to 3 was specific, since all substances influenced neither hepatitis C virus (HCV) nor HIV-1 replication. Molecular docking suggests binding at a specific allosteric binding site. In addition to the in vitro assays, a cell-based PR assay was developed to test these substances in a replication-independent way. The new compounds inhibited the DENV PR with IC50s in the low-micromolar or submicromolar range in cells. Furthermore, these novel PIs inhibit viral replication at submicromolar concentrations. PMID:25487800

  17. Monoclonal antibodies against NS1 protein of Goose parvovirus.

    PubMed

    Qiu, Zheng; Tian, Wei; Yu, Tianfei; Li, Li; Ma, Bo; Wang, Junwei

    2012-04-01

    In the present study, monoclonal antibodies (MAbs) against NS1 protein of Goose parvovirus (GPV) were generated. The secreted MAbs were obtained by fusing mouse myeloma cells and spleen cells of BALB/c mice, which were immunized with the plasmid pcDNA3.1-GPV-NS1 and recombinant protein of GPV-NS1. With indirect ELISA, six hybridoma cell lines against GPV-NS1 were screened. The subtypes of the two MAbs were IgG2a; the others were IgM. The light chain was κ. Western blot analysis showed that six MAbs reacted with recombinant protein GPV-NS1. GPV-NS1 was dissected into 15 overlapping epitopes, which were used to react with MAbs in Western blot. Results showed that six MAbs recognized NS1 protein linear B-cell epitopes located at the C-terminus 453-514 aa, 485-542 aa, and 533-598 aa.

  18. Psammaplin A inhibits hepatitis C virus NS3 helicase.

    PubMed

    Salam, Kazi Abdus; Furuta, Atsushi; Noda, Naohiro; Tsuneda, Satoshi; Sekiguchi, Yuji; Yamashita, Atsuya; Moriishi, Kohji; Nakakoshi, Masamichi; Tsubuki, Masayoshi; Tani, Hidenori; Tanaka, Junichi; Akimitsu, Nobuyoshi

    2013-10-01

    Hepatitis C virus (HCV) is the causative agent of hepatitis C, a chronic infectious disease that can lead to development of hepatocellular carcinoma. The NS3 nucleoside triphosphatase (NTPase)/helicase has an essential role in HCV replication, and is therefore an attractive target for direct-acting antiviral strategies. In this study, we employed high-throughput screening using a photo-induced electron transfer (PET) system to identify an inhibitor of NS3 helicase from marine organism extracts. We successfully identified psammaplin A as a novel NS3 inhibitor. The dose-response relationship clearly demonstrates the inhibition of NS3 RNA helicase and ATPase activities by psammaplin A, with IC₅₀ values of 17 and 32 μM, respectively. Psammaplin A has no influence on the apparent Km value (0.4 mM) of NS3 ATPase activity, and acts as a non-competitive inhibitor. Additionally, it inhibits the binding of NS3 to single-stranded RNA in a dose-dependent manner. Furthermore, psammaplin A shows an inhibitory effect on viral replication, with EC₅₀ values of 6.1 and 6.3 μM in subgenomic replicon cells derived from genotypes 1b and 2a, respectively. We postulate that psammaplin A is a potential anti-viral agent through the inhibition of ATPase, RNA binding and helicase activities of NS3. PMID:23359228

  19. Silencing by H-NS Potentiated the Evolution of Salmonella

    PubMed Central

    Rao, Chitong; Leung, Andrea S.; Ngai, David Hon-Man; Ensminger, Alexander W.; Navarre, William Wiley

    2014-01-01

    The bacterial H-NS protein silences expression from sequences with higher AT-content than the host genome and is believed to buffer the fitness consequences associated with foreign gene acquisition. Loss of H-NS results in severe growth defects in Salmonella, but the underlying reasons were unclear. An experimental evolution approach was employed to determine which secondary mutations could compensate for the loss of H-NS in Salmonella. Six independently derived S. Typhimurium hns mutant strains were serially passaged for 300 generations prior to whole genome sequencing. Growth rates of all lineages dramatically improved during the course of the experiment. Each of the hns mutant lineages acquired missense mutations in the gene encoding the H-NS paralog StpA encoding a poorly understood H-NS paralog, while 5 of the mutant lineages acquired deletions in the genes encoding the Salmonella Pathogenicity Island-1 (SPI-1) Type 3 secretion system critical to invoke inflammation. We further demonstrate that SPI-1 misregulation is a primary contributor to the decreased fitness in Salmonella hns mutants. Three of the lineages acquired additional loss of function mutations in the PhoPQ virulence regulatory system. Similarly passaged wild type Salmonella lineages did not acquire these mutations. The stpA missense mutations arose in the oligomerization domain and generated proteins that could compensate for the loss of H-NS to varying degrees. StpA variants most able to functionally substitute for H-NS displayed altered DNA binding and oligomerization properties that resembled those of H-NS. These findings indicate that H-NS was central to the evolution of the Salmonellae by buffering the negative fitness consequences caused by the secretion system that is the defining characteristic of the species. PMID:25375226

  20. The influenza virus NS1 protein as a therapeutic target.

    PubMed

    Engel, Daniel A

    2013-09-01

    Nonstructural protein 1 (NS1) of influenza A virus plays a central role in virus replication and blockade of the host innate immune response, and is therefore being considered as a potential therapeutic target. The primary function of NS1 is to dampen the host interferon (IFN) response through several distinct molecular mechanisms that are triggered by interactions with dsRNA or specific cellular proteins. Sequestration of dsRNA by NS1 results in inhibition of the 2'-5' oligoadenylate synthetase/RNase L antiviral pathway, and also inhibition of dsRNA-dependent signaling required for new IFN production. Binding of NS1 to the E3 ubiquitin ligase TRIM25 prevents activation of RIG-I signaling and subsequent IFN induction. Cellular RNA processing is also targeted by NS1, through recognition of cleavage and polyadenylation specificity factor 30 (CPSF30), leading to inhibition of IFN-β mRNA processing as well as that of other cellular mRNAs. In addition NS1 binds to and inhibits cellular protein kinase R (PKR), thus blocking an important arm of the IFN system. Many additional proteins have been reported to interact with NS1, either directly or indirectly, which may serve its anti-IFN and additional functions, including the regulation of viral and host gene expression, signaling pathways and viral pathogenesis. Many of these interactions are potential targets for small-molecule intervention. Structural, biochemical and functional studies have resulted in hypotheses for drug discovery approaches that are beginning to bear experimental fruit, such as targeting the dsRNA-NS1 interaction, which could lead to restoration of innate immune function and inhibition of virus replication. This review describes biochemical, cell-based and nucleic acid-based approaches to identifying NS1 antagonists.

  1. The influenza virus NS1 protein as a therapeutic target

    PubMed Central

    Engel, Daniel A.

    2015-01-01

    Nonstructural protein 1 (NS1) of influenza A virus plays a central role in virus replication and blockade of the host innate immune response, and is therefore being considered as a potential therapeutic target. The primary function of NS1 is to dampen the host interferon (IFN) response through several distinct molecular mechanisms that are triggered by interactions with dsRNA or specific cellular proteins. Sequestration of dsRNA by NS1 results in inhibition of the 2’-5’ oligoadenylate synthetase/RNase L antiviral pathway, and also inhibition of dsRNA-dependent signaling required for new IFN production. Binding of NS1 to the E3 ubiquitin ligase TRIM25 prevents activation of RIG-I signaling and subsequent IFN induction. Cellular RNA processing is also targeted by NS1, through recognition of cleavage and polyadenylation specificity factor 30 (CPSF30), leading to inhibition of IFN- mRNA processing as well as that of other cellular mRNAs. In addition NS1 binds to and inhibits cellular protein kinase R (PKR), thus blocking an important arm of the IFN system. Many additional proteins have been reported to interact with NS1, either directly or indirectly, which may serve its anti-IFN and additional functions, including the regulation of viral and host gene expression, signaling pathways and viral pathogenesis. Many of these interactions are potential targets for small-molecule intervention. Structural, biochemical and functional studies have resulted in hypotheses for drug discovery approaches that are beginning to bear experimental fruit, such as targeting the dsRNA-NS1 interaction, which could lead to restoration of innate immune function and inhibition of virus replication. This review describes biochemical, cell-based and nucleic acid-based approaches to identifying NS1 antagonists. PMID:23796981

  2. Differential Rotation via Tracking of Coronal Bright Points.

    NASA Astrophysics Data System (ADS)

    McAteer, James; Boucheron, Laura E.; Osorno, Marcy

    2016-05-01

    The accurate computation of solar differential rotation is important both as a constraint for, and evidence towards, support of models of the solar dynamo. As such, the use of Xray and Extreme Ultraviolet bright points to elucidate differential rotation has been studied in recent years. In this work, we propose the automated detection and tracking of coronal bright points (CBPs) in a large set of SDO data for re-evaluation of solar differential rotation and comparison to other results. The big data aspects, and high cadence, of SDO data mitigate a few issues common to detection and tracking of objects in image sequences and allow us to focus on the use of CBPs to determine differential rotation. The high cadence of the data allows to disambiguate individual CBPs between subsequent images by allowing for significant spatial overlap, i.e., by the fact that the CBPs will rotate a short distance relative to their size. The significant spatial overlap minimizes the effects of incorrectly detected CBPs by reducing the occurrence of outlier values of differential rotation. The big data aspects of the data allows to be more conservative in our detection of CBPs (i.e., to err on the side of missing CBPs rather than detecting extraneous CBPs) while still maintaining statistically larger populations over which to study characteristics. The ability to compute solar differential rotation through the automated detection and tracking of a large population of CBPs will allow for further analyses such as the N-S asymmetry of differential rotation, variation of differential rotation over the solar cycle, and a detailed study of the magnetic flux underlying the CBPs.

  3. The Influenza A Virus Protein NS1 Displays Structural Polymorphism

    PubMed Central

    Carrillo, Berenice; Choi, Jae-Mun; Bornholdt, Zachary A.; Sankaran, Banumathi; Rice, Andrew P.

    2014-01-01

    ABSTRACT NS1 of influenza A virus is a potent antagonist of host antiviral interferon responses. This multifunctional protein with two distinctive domains, an RNA-binding domain (RBD) and an effector domain (ED) separated by a linker region (LR), is implicated in replication, pathogenesis, and host range. Although the structures of individual domains of NS1 from different strains of influenza viruses have been reported, the only structure of full-length NS1 available to date is from an H5N1 strain (A/Vietnam/1203/2004). By carrying out crystallographic analyses of full-length H6N6-NS1 (A/blue-winged teal/MN/993/1980) and an LR deletion mutant, combined with mutational analysis, we show here that these full-length NS1 structures provide an exquisite structural sampling of various conformational states of NS1 that based on the orientation of the ED with respect to RBD can be summarized as “open,” “semi-open,” and “closed” conformations. Our studies show that preference for these states is clearly dictated by determinants such as linker length, residue composition at position 71, and a mechanical hinge, providing a structural basis for strain-dependent functional variations in NS1. Because of the flexibility inherent in the LR, any particular NS1 could sample the conformational space around these states to engage ED in different quaternary interactions so that it may participate in specific protein-protein or protein-RNA interactions to allow for the known multifunctionality of NS1. We propose that such conformational plasticity provides a mechanism for autoregulating NS1 functions, depending on its temporal distribution, posttranslational modifications, and nuclear or cellular localization, during the course of virus infection. IMPORTANCE NS1 of influenza A virus is a multifunctional protein associated with numerous strain-specific regulatory functions during viral infection, including conferring resistance to antiviral interferon induction, replication

  4. Just How Bright Is a Laser?

    ERIC Educational Resources Information Center

    Van Baak, David A.

    1995-01-01

    Attempts to quantify the subjective sensation of brightness of the spot projected by a helium-neon laser and compares this with conventional sources of light. Provides an exercise in using the blackbody radiation formulas. (JRH)

  5. New Observations of Subarcsecond Photospheric Bright Points

    NASA Technical Reports Server (NTRS)

    Berger, T. E.; Schrijver, C. J.; Shine, R. A.; Tarbell, T. D.; Title, A. M.; Scharmer, G.

    1995-01-01

    We have used an interference filter centered at 4305 A within the bandhead of the CH radical (the 'G band') and real-time image selection at the Swedish Vacuum Solar Telescope on La Palma to produce very high contrast images of subarcsecond photospheric bright points at all locations on the solar disk. During the 6 day period of 1993 September 15-20 we observed active region NOAA 7581 from its appearance on the East limb to a near-disk-center position on September 20. A total of 1804 bright points were selected for analysis from the disk center image using feature extraction image processing techniques. The measured Full Width at Half Maximum (FWHM) distribution of the bright points in the image is lognormal with a modal value of 220 km (0 sec .30) and an average value of 250 km (0 sec .35). The smallest measured bright point diameter is 120 km (0 sec .17) and the largest is 600 km (O sec .69). Approximately 60% of the measured bright points are circular (eccentricity approx. 1.0), the average eccentricity is 1.5, and the maximum eccentricity corresponding to filigree in the image is 6.5. The peak contrast of the measured bright points is normally distributed. The contrast distribution variance is much greater than the measurement accuracy, indicating a large spread in intrinsic bright-point contrast. When referenced to an averaged 'quiet-Sun' area in the image, the modal contrast is 29% and the maximum value is 75%; when referenced to an average intergranular lane brightness in the image, the distribution has a modal value of 61% and a maximum of 119%. The bin-averaged contrast of G-band bright points is constant across the entire measured size range. The measured area of the bright points, corrected for pixelation and selection effects, covers about 1.8% of the total image area. Large pores and micropores occupy an additional 2% of the image area, implying a total area fraction of magnetic proxy features in the image of 3.8%. We discuss the implications of this

  6. New Observations of Subarcsecond Photospheric Bright Points

    NASA Technical Reports Server (NTRS)

    Berger, T. E.; Schrijver, C. J.; Shine, R. A.; Tarbell, T. D.; Title, A. M.; Scharmer, G.

    1995-01-01

    We have used an interference filter centered at 4305 A within the bandhead of the CH radical (the 'G band') and real-time image selection at the Swedish Vacuum Solar Telescope on La Palma to produce very high contrast images of subarcsecond photospheric bright points at all locations on the solar disk. During the 6 day period of 15-20 Sept. 1993 we observed active region NOAA 7581 from its appearance on the East limb to a near-disk-center position on 20 Sept. A total of 1804 bright points were selected for analysis from the disk center image using feature extraction image processing techniques. The measured FWHM distribution of the bright points in the image is lognormal with a modal value of 220 km (0.30 sec) and an average value of 250 km (0.35 sec). The smallest measured bright point diameter is 120 km (0.17 sec) and the largest is 600 km (O.69 sec). Approximately 60% of the measured bright points are circular (eccentricity approx. 1.0), the average eccentricity is 1.5, and the maximum eccentricity corresponding to filigree in the image is 6.5. The peak contrast of the measured bright points is normally distributed. The contrast distribution variance is much greater than the measurement accuracy, indicating a large spread in intrinsic bright-point contrast. When referenced to an averaged 'quiet-Sun' area in the image, the modal contrast is 29% and the maximum value is 75%; when referenced to an average intergranular lane brightness in the image, the distribution has a modal value of 61% and a maximum of 119%. The bin-averaged contrast of G-band bright points is constant across the entire measured size range. The measured area of the bright points, corrected for pixelation and selection effects, covers about 1.8% of the total image area. Large pores and micropores occupy an additional 2% of the image area, implying a total area fraction of magnetic proxy features in the image of 3.8%. We discuss the implications of this area fraction measurement in the context of

  7. Several evolutionary channels for bright planetary nebulae

    NASA Astrophysics Data System (ADS)

    Richer, Michael G.; McCall, Marshall L.

    2016-08-01

    The populations of bright planetary nebulae in the discs of spirals appear to differ in their spectral properties from those in ellipticals and the bulges of spirals. The bright planetary nebulae from the bulge of the Milky Way are entirely compatible with those observed in the discs of spiral galaxies. The similarity might be explained if the bulge of the Milky Way evolved secularly from the disc, in which case the bulge should be regarded as a pseudo-bulge.

  8. Evolution of bulgeless low surface brightness galaxies

    NASA Astrophysics Data System (ADS)

    Shao, Xu; Hammer, Francois; Yang, Yanbin; Liang, Yanchun

    2015-08-01

    We study the environment, the morphology and stellar population of bulgeless low surface brightness (LSB) galaxies in a volume-limited sample. The differences of environments between LSB and high surface brightness (HSB) galaxies are not obvious, suggesting they may locate in similar environments. The stellar populations of LSB galaxies in low density region are similar with those of LSB galaxies in high density region. Irregular LSB galaxies have more young stars and are more metal-poor than the regular LSB galaxies.

  9. Observations and diagnostics in high brightness beams

    NASA Astrophysics Data System (ADS)

    Cianchi, A.; Anania, M. P.; Bisesto, F.; Castellano, M.; Chiadroni, E.; Pompili, R.; Shpakov, V.

    2016-09-01

    The brightness is a figure of merit largely used in the light sources, like FEL (Free Electron Lasers), but it is also fundamental in several other applications, as for instance Compton backscattering sources, beam driven plasma accelerators and THz sources. Advanced diagnostics are essential tools in the development of high brightness beams. 6D electron beam diagnostics will be reviewed with emphasis on emittance measurement.

  10. Determining the bivariate brightness distribution of galaxies.

    NASA Astrophysics Data System (ADS)

    Boyce, P. J.; Phillipps, S.

    1995-04-01

    In this paper we describe a set of criteria which we propose a sample of galaxies must satisfy if it is to be useful for determining the bivariate brightness distribution (BBD) of galaxies in luminosity and surface brightness and we consider the prospects for deriving such a sample. First, we note that determinations of the galaxy luminosity function can be seriously in error if surface brightness (visibility) selection effects are ignored. We suggest that a determination of the BBD is a more physically useful aim. A straightforward way to obtain the BBD would be to determine a luminosity function in a set of narrow surface brightness bins. We propose a set of criteria which the sample of galaxies in each surface brightness bin must satisfy if it is to be reliably used in such a determination. Each sample should be restricted to a well defined range in morphological type, the measured isophotal size and magnitude and the surface brightness of each galaxy should be corrected to a common galactic inclination, all galaxies should have measured redshifts and the sample should be complete to a known isophotal size and/or magnitude. We then describe a rigorous method for selecting samples which satisfy these criteria from existing catalogues of galaxies. We apply this method to the ESO-LV catalogue and find that from the intial sample of 11000 galaxies with a disk component we can only find 5 subsamples in half-magnitude wide surface brightness bins which satisfy our proposed criteria. The largest derived subsample contains only 27 galaxies, far too few to determine a luminosity function at its surface brightness. We suggest that had our proposed criteria been applied to the samples used in previous determinations of the BBD or the galaxy luminosity function then sample sizes would have been greatly reduced. For this reason, we suggest that the conclusions of previous work should be treated with caution.

  11. Monoclonal Antibodies Against NS2B of Japanese Encephalitis Virus.

    PubMed

    Dong, Qian; Xu, Qiuping; Ruan, Xindi; Huang, Shaomei; Cao, Shengbo

    2015-04-01

    Japanese encephalitis (JE) is one of the most important viral encephalitis, caused by the Japanese encephalitis virus (JEV). The function of non-structural protein 2B (NS2B) mostly remains unclear. In our study, NS2B of Japanese encephalitis virus (JEV) was expressed in Escherichia coli and purified by dialysis. After fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from NS2B protein immunized mice, three clones of monoclonal antibodies (MAbs), named 1B9, 3E12, and 4E6, were generated. The specificity and sensitivity of MAbs were demonstrated by ELISA, indirect immunofluorescence assay, and Western blot. These MAbs will be useful in further exploration of the functions of NS2B and the pathogenesis of Japanese encephalitis virus. PMID:25897607

  12. NS1: A corner piece in the dengue pathogenesis puzzle?

    PubMed

    Thomas, Stephen J

    2015-09-01

    Soluble dengue virus NS1 protein induces proinflammatory immune responses via Toll-like receptor 4 and disrupts endothelial cell integrity, resulting in vascular leakage (Beatty et al. and Modhiran et al., this issue). PMID:26355028

  13. Flavivirus NS1: a multifaceted enigmatic viral protein.

    PubMed

    Rastogi, Meghana; Sharma, Nikhil; Singh, Sunit Kumar

    2016-01-01

    Flaviviruses are emerging arthropod-borne viruses representing an immense global health problem. The prominent viruses of this group include dengue virus, yellow fever virus, Japanese encephalitis virus, West Nile virus tick borne encephalitis virus and Zika Virus. These are endemic in many parts of the world. They are responsible for the illness ranging from mild flu like symptoms to severe hemorrhagic, neurologic and cognitive manifestations leading to death. NS1 is a highly conserved non-structural protein among flaviviruses, which exist in diverse forms. The intracellular dimer form of NS1 plays role in genome replication, whereas, the secreted hexamer plays role in immune evasion. The secreted NS1 has been identified as a potential diagnostic marker for early detection of the infections caused by flaviviruses. In addition to the diagnostic marker, the importance of NS1 has been reported in the development of therapeutics. NS1 based subunit vaccines are at various stages of development. The structural details and diverse functions of NS1 have been discussed in detail in this review.

  14. Flavivirus NS1: a multifaceted enigmatic viral protein.

    PubMed

    Rastogi, Meghana; Sharma, Nikhil; Singh, Sunit Kumar

    2016-01-01

    Flaviviruses are emerging arthropod-borne viruses representing an immense global health problem. The prominent viruses of this group include dengue virus, yellow fever virus, Japanese encephalitis virus, West Nile virus tick borne encephalitis virus and Zika Virus. These are endemic in many parts of the world. They are responsible for the illness ranging from mild flu like symptoms to severe hemorrhagic, neurologic and cognitive manifestations leading to death. NS1 is a highly conserved non-structural protein among flaviviruses, which exist in diverse forms. The intracellular dimer form of NS1 plays role in genome replication, whereas, the secreted hexamer plays role in immune evasion. The secreted NS1 has been identified as a potential diagnostic marker for early detection of the infections caused by flaviviruses. In addition to the diagnostic marker, the importance of NS1 has been reported in the development of therapeutics. NS1 based subunit vaccines are at various stages of development. The structural details and diverse functions of NS1 have been discussed in detail in this review. PMID:27473856

  15. Energy-exchange collisions of dark-bright-bright vector solitons

    NASA Astrophysics Data System (ADS)

    Radhakrishnan, R.; Manikandan, N.; Aravinthan, K.

    2015-12-01

    We find a dark component guiding the practically interesting bright-bright vector one-soliton to two different parametric domains giving rise to different physical situations by constructing a more general form of three-component dark-bright-bright mixed vector one-soliton solution of the generalized Manakov model with nine free real parameters. Moreover our main investigation of the collision dynamics of such mixed vector solitons by constructing the multisoliton solution of the generalized Manakov model with the help of Hirota technique reveals that the dark-bright-bright vector two-soliton supports energy-exchange collision dynamics. In particular the dark component preserves its initial form and the energy-exchange collision property of the bright-bright vector two-soliton solution of the Manakov model during collision. In addition the interactions between bound state dark-bright-bright vector solitons reveal oscillations in their amplitudes. A similar kind of breathing effect was also experimentally observed in the Bose-Einstein condensates. Some possible ways are theoretically suggested not only to control this breathing effect but also to manage the beating, bouncing, jumping, and attraction effects in the collision dynamics of dark-bright-bright vector solitons. The role of multiple free parameters in our solution is examined to define polarization vector, envelope speed, envelope width, envelope amplitude, grayness, and complex modulation of our solution. It is interesting to note that the polarization vector of our mixed vector one-soliton evolves in sphere or hyperboloid depending upon the initial parametric choices.

  16. Energy-exchange collisions of dark-bright-bright vector solitons.

    PubMed

    Radhakrishnan, R; Manikandan, N; Aravinthan, K

    2015-12-01

    We find a dark component guiding the practically interesting bright-bright vector one-soliton to two different parametric domains giving rise to different physical situations by constructing a more general form of three-component dark-bright-bright mixed vector one-soliton solution of the generalized Manakov model with nine free real parameters. Moreover our main investigation of the collision dynamics of such mixed vector solitons by constructing the multisoliton solution of the generalized Manakov model with the help of Hirota technique reveals that the dark-bright-bright vector two-soliton supports energy-exchange collision dynamics. In particular the dark component preserves its initial form and the energy-exchange collision property of the bright-bright vector two-soliton solution of the Manakov model during collision. In addition the interactions between bound state dark-bright-bright vector solitons reveal oscillations in their amplitudes. A similar kind of breathing effect was also experimentally observed in the Bose-Einstein condensates. Some possible ways are theoretically suggested not only to control this breathing effect but also to manage the beating, bouncing, jumping, and attraction effects in the collision dynamics of dark-bright-bright vector solitons. The role of multiple free parameters in our solution is examined to define polarization vector, envelope speed, envelope width, envelope amplitude, grayness, and complex modulation of our solution. It is interesting to note that the polarization vector of our mixed vector one-soliton evolves in sphere or hyperboloid depending upon the initial parametric choices.

  17. Replacement of the respiratory syncytial virus nonstructural proteins NS1 and NS2 by the V protein of parainfluenza virus 5

    SciTech Connect

    Tran, Kim C.; He, Biao; Teng, Michael N.

    2007-11-10

    Paramyxoviruses have been shown to produce proteins that inhibit interferon production and signaling. For human respiratory syncytial virus (RSV), the nonstructural NS1 and NS2 proteins have been shown to have interferon antagonist activity through an unknown mechanism. To understand further the functions of NS1 and NS2, we generated recombinant RSV in which both NS1 and NS2 were replaced by the PIV5 V protein, which has well-characterized IFN antagonist activities ({delta}NS1/2-V). Expression of V was able to partially inhibit IFN responses in {delta}NS1/2-V-infected cells. In addition, the replication kinetics of {delta}NS1/2-V were intermediate between {delta}NS1/2 and wild-type (rA2) in A549 cells. However, expression of V did not affect the ability of {delta}NS1/2-V to activate IRF3 nuclear translocation and IFN{beta} transcription. These data indicate that V was able to replace some of the IFN inhibitory functions of the RSV NS1 and NS2 proteins, but also that NS1 and NS2 have functions in viral replication beyond IFN antagonism.

  18. Microwave Brightness Temperatures of Tilted Convective Systems

    NASA Technical Reports Server (NTRS)

    Hong, Ye; Haferman, Jeffrey L.; Olson, William S.; Kummerow, Christian D.

    1998-01-01

    Aircraft and ground-based radar data from the Tropical Ocean and Global Atmosphere Coupled-Ocean Atmosphere Response Experiment (TOGA COARE) show that convective systems are not always vertical. Instead, many are tilted from vertical. Satellite passive microwave radiometers observe the atmosphere at a viewing angle. For example, the Special Sensor Microwave/Imager (SSM/I) on Defense Meteorological Satellite Program (DMSP) satellites and the Tropical Rainfall Measurement Mission (TRMM) Microwave Imager (TMI) on the TRMM satellite have an incident angle of about 50deg. Thus, the brightness temperature measured from one direction of tilt may be different than that viewed from the opposite direction due to the different optical depth. This paper presents the investigation of passive microwave brightness temperatures of tilted convective systems. To account for the effect of tilt, a 3-D backward Monte Carlo radiative transfer model has been applied to a simple tilted cloud model and a dynamically evolving cloud model to derive the brightness temperature. The radiative transfer results indicate that brightness temperature varies when the viewing angle changes because of the different optical depth. The tilt increases the displacements between high 19 GHz brightness temperature (Tb(sub 19)) due to liquid emission from lower level of cloud and the low 85 GHz brightness temperature (Tb(sub 85)) due to ice scattering from upper level of cloud. As the resolution degrades, the difference of brightness temperature due to the change of viewing angle decreases dramatically. The dislocation between Tb(sub 19) and Tb(sub 85), however, remains prominent.

  19. The Sky Brightness Data Archive (SBDA)

    NASA Astrophysics Data System (ADS)

    Craine, Eric R.; Craine, Erin M.; Craine, Brian L.

    2011-05-01

    Although many astronomers have long been sensitive to issues of light pollution and deteriorating sky quality it is only in recent years that such interest has extended to other groups including, among others, ecologists, health professionals, and urban planners. Issues of light pollution and loss of dark skies are starting to appear in the scientific literature in the context of health and behavior impacts on both human and animal life. Nonetheless, a common deficiency in most such studies is the absence of historical or baseline data against which to compare sky brightness trends and temporal changes. To address this deficiency we have begun to collect a variety of types of quantitative sky brightness data for insertion in an international sky brightness archive that can be accessed for research projects which are dependent upon an understanding of the nature of local light pollution issues. To aid this process we have developed a mobile sky brightness meter which automatically logs sky brightness and observation location. The device can be stationary for long periods of time or can be easily transported for continuous sky brightness measurement from ground vehicles, boats, or aircraft. The sampling rate is typically about 0.25Hz. We present here examples of different modes of sky brightness measurement, various means of displaying and analyzing such data, ways to interpret natural astronomical phenomena apparent in the data, and suggest a number of complementary scientific projects that may capture the interest of both professional and amateur scientists. Finally, we discuss the status of the archive and ways that potential contributors may submit their observations for publication in the archive.

  20. Proper processing of dengue virus nonstructural glycoprotein NS1 requires the N-terminal hydrophobic signal sequence and the downstream nonstructural protein NS2a.

    PubMed

    Falgout, B; Chanock, R; Lai, C J

    1989-05-01

    Expression of dengue virus gene products involves specific proteolytic cleavages of a precursor polyprotein. To study the flanking sequences required for expression of the dengue virus nonstructural glycoprotein NS1, we constructed a series of recombinant vaccinia viruses that contain the coding sequence for NS1 in combination with various lengths of upstream and downstream sequences. The NS1 products expressed by these viruses in infected CV-1 cells were immune precipitated and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The data show that the 24-residue hydrophobic sequence preceding NS1 was necessary and sufficient for the production of glycosylated NS1 and that this sequence was cleaved from NS1 in the absence of most dengue virus proteins. This finding is consistent with previous proposals that this hydrophobic sequence serves as an N-terminal signal sequence that is cleaved by signal peptidase. The cleavage between the C terminus of NS1 and the downstream protein NS2a occurred when the complete NS2a was present. Recombinant viruses containing NS1 plus 15 or 49% of NS2a produced proteins larger than authentic NS1, indicating that the cleavage between NS1 and NS2a had not occurred. Failure of cleavage was not corrected by coinfection with a recombinant virus capable of cleavage. These results suggest that NS2a may be a cis-acting protease that cleaves itself from NS1, or NS2a may provide sequences for recognition by a specific cellular protease that cleaves at the NS1-NS2a junction.

  1. The NS1 protein: a multitasking virulence factor.

    PubMed

    Ayllon, Juan; García-Sastre, Adolfo

    2015-01-01

    The non-structural protein 1 of influenza virus (NS1) is a relatively small polypeptide with an outstanding number of ascribed functions. NS1 is the main viral antagonist of the innate immune response during influenza virus infection, chiefly by inhibiting the type I interferon system at multiple steps. As such, its role is critical to overcome the first barrier the host presents to halt the viral infection. However, the pro-viral activities of this well-studied protein go far beyond and include regulation of viral RNA and protein synthesis, and disruption of the host cell homeostasis by dramatically affecting general gene expression while tweaking the PI3K signaling network. Because of all of this, NS1 is a key virulence factor that impacts influenza pathogenesis, and adaptation to new hosts, making it an attractive target for control strategies. Here, we will overview the many roles that have been ascribed to the NS1 protein, and give insights into the sequence features and structural properties that make them possible, highlighting the need to understand how NS1 can actually perform all of these functions during viral infection. PMID:25007846

  2. The NS1 protein: a multitasking virulence factor.

    PubMed

    Ayllon, Juan; García-Sastre, Adolfo

    2015-01-01

    The non-structural protein 1 of influenza virus (NS1) is a relatively small polypeptide with an outstanding number of ascribed functions. NS1 is the main viral antagonist of the innate immune response during influenza virus infection, chiefly by inhibiting the type I interferon system at multiple steps. As such, its role is critical to overcome the first barrier the host presents to halt the viral infection. However, the pro-viral activities of this well-studied protein go far beyond and include regulation of viral RNA and protein synthesis, and disruption of the host cell homeostasis by dramatically affecting general gene expression while tweaking the PI3K signaling network. Because of all of this, NS1 is a key virulence factor that impacts influenza pathogenesis, and adaptation to new hosts, making it an attractive target for control strategies. Here, we will overview the many roles that have been ascribed to the NS1 protein, and give insights into the sequence features and structural properties that make them possible, highlighting the need to understand how NS1 can actually perform all of these functions during viral infection.

  3. A selective deficit in the appreciation and recognition of brightness: brightness agnosia?

    PubMed

    Nijboer, Tanja C W; Nys, Gudrun M S; van der Smagt, Maarten J; de Haan, Edward H F

    2009-01-01

    We report a patient with extensive brain damage in the right hemisphere who demonstrated a severe impairment in the appreciation of brightness. Acuity, contrast sensitivity as well as luminance discrimination were normal, suggesting her brightness impairment is not a mere consequence of low-level sensory impairments. The patient was not able to indicate the darker or the lighter of two grey squares, even though she was able to see that they differed. In addition, she could not indicate whether the lights in a room were switched on or off, nor was she able to differentiate between normal greyscale images and inverted greyscale images. As the patient recognised objects, colours, and shapes correctly, the impairment is specific for brightness. As low-level, sensory processing is normal, this specific deficit in the recognition and appreciation of brightness appears to be of a higher, cognitive level, the level of semantic knowledge. This appears to be the first report of 'brightness agnosia'.

  4. Brightness discrimination in budgerigars (Melopsittacus undulatus).

    PubMed

    Lind, Olle; Karlsson, Sandra; Kelber, Almut

    2013-01-01

    Birds have excellent spatial acuity and colour vision compared to other vertebrates while spatial contrast sensitivity is relatively poor for unknown reasons. Contrast sensitivity describes the detection of gratings of varying spatial frequency. It is unclear whether bird brightness discrimination between large uniform fields is poor as well. Here we show that budgerigars (Melopsittacus undulatus) need a Michelson contrast of 0.09 to discriminate between large spatially separated achromatic fields in bright light conditions. This is similar to the peak contrast sensitivity of 10.2 (0.098 Michelson contrast) for achromatic grating stimuli established in earlier studies. The brightness discrimination threshold described in Weber fractions is 0.18, which is modest compared to other vertebrates. PMID:23349946

  5. High-brightness electron injectors: A review

    SciTech Connect

    Sheffield, R.L.

    1989-01-01

    The last decade has seen increased emphasis on the development of high-brightness electron beams because of rigorous requirements of the new generation of colliders and the advent of free-electron lasers. This talk describes the approaches now being explored for attaining intense, bright electron beams. The methods for producing bright electron beams include photocathode-based, short-pulse injectors; dc electrostatic accelerator sources; long-pulse beams, which are then compressed in time using subharmonic bunching; combining first and third harmonics in an accelerator to attain the equivalent of high-gradient dc fields; and LaB/sub 6/ rf guns. For several of the approaches, the temporal length of the electron pulse is decreased after acceleration to relativistic energies by impressing an energy spread on the electron bunch and using a nonisochronous beam-transport system to increase the peak current. 37 refs., 7 figs.

  6. A brightness exceeding simulated Langmuir limit

    SciTech Connect

    Nakasuji, Mamoru

    2013-08-15

    When an excitation of the first lens determines a beam is parallel beam, a brightness that is 100 times higher than Langmuir limit is measured experimentally, where Langmuir limits are estimated using a simulated axial cathode current density which is simulated based on a measured emission current. The measured brightness is comparable to Langmuir limit, when the lens excitation is such that an image position is slightly shorter than a lens position. Previously measured values of brightness for cathode apical radii of curvature 20, 60, 120, 240, and 480 μm were 8.7, 5.3, 3.3, 2.4, and 3.9 times higher than their corresponding Langmuir limits, respectively, in this experiment, the lens excitation was such that the lens and the image positions were 180 mm and 400 mm, respectively. From these measured brightness for three different lens excitation conditions, it is concluded that the brightness depends on the first lens excitation. For the electron gun operated in a space charge limited condition, some of the electrons emitted from the cathode are returned to the cathode without having crossed a virtual cathode. Therefore, method that assumes a Langmuir limit defining method using a Maxwellian distribution of electron velocities may need to be revised. For the condition in which the values of the exceeding the Langmuir limit are measured, the simulated trajectories of electrons that are emitted from the cathode do not cross the optical axis at the crossover, thus the law of sines may not be valid for high brightness electron beam systems.

  7. The historical investigation of cometary brightness

    NASA Astrophysics Data System (ADS)

    Hughes, David W.

    1998-12-01

    The interpretation of the way in which the brightness of a comet varied as a function of both its heliocentric and geocentric distance was essentially started by Isaac Newton in his book Philosophiae Naturalis Principia Mathematica, published in 1687. Astronomers have argued about the form of this variability ever since, and for many years it was regarded as an important clue as to the physical nature of the cometary nucleus and its decay process. This paper reviews our understanding of the causes of cometary brightness variability between about 1680 and the 1950s.

  8. Evolution of bulgeless low surface brightness galaxies

    NASA Astrophysics Data System (ADS)

    Shao, X.; Hammer, F.; Yang, Y. B.; Liang, Y. C.

    Based on the Sloan Digital Sky Survey DR 7, we investigate the environment, morphology, and stellar population of bulgeless low surface-brightness (LSB) galaxies in a volume-limited sample with redshift ranging from 0.024 to 0.04 and M r <= -18.8. We find that, for bulgeless galaxies, the surface brightness does not depend on the environment. Irregular LSB galaxies have more young stars and are more metal-poor than regular LSB galaxies. These results suggest that the evolution of LSB galaxies may be driven by their dynamics, including mergers rather than by their large-scale environment.

  9. Observed brightness distributions in overcast skies.

    PubMed

    Lee, Raymond L; Devan, David E

    2008-12-01

    Beneath most overcasts, clouds' motions and rapidly changing optical depths complicate mapping their angular distributions of luminance L(v) and visible-wavelength radiance L. Fisheye images of overcast skies taken with a radiometer-calibrated digital camera provide a useful new approach to solving this problem. Maps calculated from time-averaged images of individual overcasts not only show their brightness distributions in unprecedented detail, but they also help solve a long-standing puzzle about where brightness maxima of overcasts are actually located. When combined with simulated radiance distributions from MODTRAN4, our measured radiances also let us estimate the gradients of cloud thickness observed in some overcasts.

  10. Richard Bright and his neurological studies.

    PubMed

    Pearce, J M S

    2009-01-01

    Richard Bright was one of the famous triumvirate of Guy's Hospital physicians in the Victorian era. Remembered for his account of glomerulonephritis (Bright's disease) he also made many important and original contributions to medicine and neurology. These included his work on cortical epileptogenesis, descriptions of simple partial (Jacksonian) seizures, infantile convulsions, and a variety of nervous diseases. Most notable were his reports of neurological studies including papers on traumatic tetanus, syringomyelia, arteries of the brain, contractures of spinal origin, tumours of the base of the brain, and narcolepsy. His career and these contributions are outlined.

  11. Richard Bright in Hungary: a reevaluation.

    PubMed

    Nagy, J; Sonkodi, S

    1997-01-01

    Richard Bright, the highly respected physician and nephrologist at Guy's Hospital, had a strong liking for travel. In 1815 he traveled in Hungary and made very important observations about the country. His 762-page book, entitled Travels from Vienna through Lower Hungary has detailed, sometimes appreciative, sometimes very critical remarks and comments on Hungarian history, art, archeology, religion, the situation of nationalities, education, social conditions, law, farming, and mining. The Hungarians cherished the memory of Bright's travel in their country as reflected in several papers and on two commemorative tablets recognizing him as a true and sincere friend of Hungary.

  12. Heterosandwich immunoswab assay for dengue virus Ns1 antigen detection.

    PubMed

    Ganguly, Advaita; Malabadi, Ravindra B; Loebenberg, Raimer; Suresh, Mavanur R; Sunwoo, Hoon H

    2014-01-01

    Dengue and the more severe dengue hemorrhagic fever have been a very critical public health problem globally. Millions of people especially in the tropical areas get infected with dengue. An efficient diagnostic is very important for early screening of dengue infection. In dengue-infected patients, the nonstructural protein NS1 is present on the surface of infected cells and secreted in plasma. The NS1 antigen is an important target for developing a quick diagnostic largely due to its long presence in the blood. We have developed a simple-to-use immunoswab-based diagnostic procedure employing monoclonal antibodies and the second-generation quadromas. The detection limit for NS1 has been established to be in the subnanogram range. The assay is very sensitive, has a visual end point, and also being extremely inexpensive. With this assay, screening time for a dengue-infected person would be very rapid. PMID:24211216

  13. NS1-binding protein abrogates the elevation of cell viability by the influenza A virus NS1 protein in association with CRKL

    SciTech Connect

    Miyazaki, Masaya; Nishihara, Hiroshi; Hasegawa, Hideki; Tashiro, Masato; Wang, Lei; Kimura, Taichi; Tanino, Mishie; Tsuda, Masumi; Tanaka, Shinya

    2013-11-29

    Highlights: •NS1 induced excessive phosphorylation of ERK and elevated cell viability. •NS1-BP expression and CRKL knockdown abolished survival effect of NS1. •NS1-BP and NS1 formed the complex through the interaction with CRKL-SH3(N). -- Abstract: The influenza A virus non-structural protein 1 (NS1) is a multifunctional virulence factor consisting of an RNA binding domain and several Src-homology (SH) 2 and SH3 binding motifs, which promotes virus replication in the host cell and helps to evade antiviral immunity. NS1 modulates general host cell physiology in association with various cellular molecules including NS1-binding protein (NS1-BP) and signaling adapter protein CRK-like (CRKL), while the physiological role of NS1-BP during influenza A virus infection especially in association with NS1 remains unclear. In this study, we analyzed the intracellular association of NS1-BP, NS1 and CRKL to elucidate the physiological roles of these molecules in the host cell. In HEK293T cells, enforced expression of NS1 of A/Beijing (H1N1) and A/Indonesia (H5N1) significantly induced excessive phosphorylation of ERK and elevated cell viability, while the over-expression of NS1-BP and the abrogation of CRKL using siRNA abolished such survival effect of NS1. The pull-down assay using GST-fusion CRKL revealed the formation of intracellular complexes of NS1-BP, NS1 and CRKL. In addition, we identified that the N-terminus SH3 domain of CRKL was essential for binding to NS1-BP using GST-fusion CRKL-truncate mutants. This is the first report to elucidate the novel function of NS1-BP collaborating with viral protein NS1 in modulation of host cell physiology. In addition, an alternative role of adaptor protein CRKL in association with NS1 and NS1-BP during influenza A virus infection is demonstrated.

  14. Fuel specificity of the hepatitis C virus NS3 helicase.

    PubMed

    Belon, Craig A; Frick, David N

    2009-05-15

    The hepatitis C virus (HCV) NS3 protein is a helicase capable of unwinding duplex RNA or DNA. This study uses a newly developed molecular-beacon-based helicase assay (MBHA) to investigate how nucleoside triphosphates (NTPs) fuel HCV helicase-catalyzed DNA unwinding. The MBHA monitors the irreversible helicase-catalyzed displacement of an oligonucleotide-bound molecular beacon so that rates of helicase translocation can be directly measured in real time. The MBHA reveals that HCV helicase unwinds DNA at different rates depending on the nature and concentration of NTPs in solution, such that the fastest reactions are observed in the presence of CTP followed by ATP, UTP, and GTP. 3'-Deoxy-NTPs generally support faster DNA unwinding, with dTTP supporting faster rates than any other canonical (d)NTP. The presence of an intact NS3 protease domain makes HCV helicase somewhat less specific than truncated NS3 bearing only its helicase region (NS3h). Various NTPs bind NS3h with similar affinities, but each NTP supports a different unwinding rate and processivity. Studies with NTP analogs reveal that specificity is determined by the nature of the Watson-Crick base-pairing region of the NTP base and the nature of the functional groups attached to the 2' and 3' carbons of the NTP sugar. The divalent metal bridging the NTP to NS3h also influences observed unwinding rates, with Mn(2+) supporting about 10 times faster unwinding than Mg(2+). Unlike Mg(2+), Mn(2+) does not support HCV helicase-catalyzed ATP hydrolysis in the absence of stimulating nucleic acids. Results are discussed in relation to models for how ATP might fuel the unwinding reaction.

  15. Structures of NS5 Methyltransferase from Zika Virus.

    PubMed

    Coloma, Javier; Jain, Rinku; Rajashankar, Kanagalaghatta R; García-Sastre, Adolfo; Aggarwal, Aneel K

    2016-09-20

    The Zika virus (ZIKV) poses a major public health emergency. To aid in the development of antivirals, we present two high-resolution crystal structures of the ZIKV NS5 methyltransferase: one bound to S-adenosylmethionine (SAM) and the other bound to SAM and 7-methyl guanosine diphosphate (7-MeGpp). We identify features of ZIKV NS5 methyltransferase that lend to structure-based antiviral drug discovery. Specifically, SAM analogs with functionalities on the Cβ atom of the methionine portion of the molecules that occupy the RNA binding tunnel may provide better specificity relative to human RNA methyltransferases.

  16. Structures of NS5 Methyltransferase from Zika Virus.

    PubMed

    Coloma, Javier; Jain, Rinku; Rajashankar, Kanagalaghatta R; García-Sastre, Adolfo; Aggarwal, Aneel K

    2016-09-20

    The Zika virus (ZIKV) poses a major public health emergency. To aid in the development of antivirals, we present two high-resolution crystal structures of the ZIKV NS5 methyltransferase: one bound to S-adenosylmethionine (SAM) and the other bound to SAM and 7-methyl guanosine diphosphate (7-MeGpp). We identify features of ZIKV NS5 methyltransferase that lend to structure-based antiviral drug discovery. Specifically, SAM analogs with functionalities on the Cβ atom of the methionine portion of the molecules that occupy the RNA binding tunnel may provide better specificity relative to human RNA methyltransferases. PMID:27633330

  17. hnRNP A2/B1 interacts with influenza A viral protein NS1 and inhibits virus replication potentially through suppressing NS1 RNA/protein levels and NS1 mRNA nuclear export

    SciTech Connect

    Wang, Yimeng; Zhou, Jianhong; Du, Yuchun

    2014-01-20

    The NS1 protein of influenza viruses is a major virulence factor and exerts its function through interacting with viral/cellular RNAs and proteins. In this study, we identified heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) as an interacting partner of NS1 proteins by a proteomic method. Knockdown of hnRNP A2/B1 by small interfering RNA (siRNA) resulted in higher levels of NS vRNA, NS1 mRNA, and NS1 protein in the virus-infected cells. In addition, we demonstrated that hnRNP A2/B1 proteins are associated with NS1 and NS2 mRNAs and that knockdown of hnRNP A2/B1 promotes transport of NS1 mRNA from the nucleus to the cytoplasm in the infected cells. Lastly, we showed that knockdown of hnRNP A2/B1 leads to enhanced virus replication. Our results suggest that hnRNP A2/B1 plays an inhibitory role in the replication of influenza A virus in host cells potentially through suppressing NS1 RNA/protein levels and NS1 mRNA nucleocytoplasmic translocation. - Highlights: • Cellular protein hnRNP A2/B1 interacts with influenza viral protein NS1. • hnRNP A2/B1 suppresses the levels of NS1 protein, vRNA and mRNA in infected cells. • hnRNP A2/B1 protein is associated with NS1 and NS2 mRNAs. • hnRNP A2/B1 inhibits the nuclear export of NS1 mRNAs. • hnRNP A2/B1 inhibits influenza virus replication.

  18. Facilitation of cell adhesion by immobilized dengue viral nonstructural protein 1 (NS1): arginine-glycine-aspartic acid structural mimicry within the dengue viral NS1 antigen.

    PubMed

    Chang, Hsin-Hou; Shyu, Huey-Fen; Wang, Yo-Ming; Sun, Der-Shan; Shyu, Rong-Hwa; Tang, Shiao-Shek; Huang, Yao-Shine

    2002-09-15

    Dengue virus infection causes life-threatening hemorrhagic fever. Increasing evidence implies that dengue viral nonstructural protein 1 (NS1) exhibits a tendency to elicit potentially hazardous autoantibodies, which show a wide spectrum of specificity against extracellular matrix and platelet antigens. How NS1 elicits autoantibodies remains unclear. To address the hypothesis that NS1 and matrix proteins may have structural and functional similarity, cell-matrix and cell-NS1 interactions were evaluated using a cell-adhesion assay. The present study showed that dengue NS1 immobilized on coverslips resulted in more cell adhesion than did the control proteins. This cell adhesion was inhibited by peptides containing arginine-glycine-aspartic acid (RGD), a motif important for integrin-mediated cell adhesion. In addition, anti-NS1 antibodies blocked RGD-mediated cell adhesion. Although there is no RGD motif in the NS1 protein sequence, these data indicate that RGD structural mimicry exists within the NS1 antigen.

  19. Brightness of synchrotron radiation from wigglers

    NASA Astrophysics Data System (ADS)

    Geloni, Gianluca; Kocharyan, Vitali; Saldin, Evgeni

    2016-01-01

    According to the literature, while calculating the brightness of synchrotron radiation from wigglers, one needs to account for the so-called 'depth-of-field' effects. In fact, the particle beam cross-section varies along the wiggler. It is usually stated that the effective photon source size increases accordingly, while the brightness is reduced. Here we claim that this is a misconception originating from an analysis of the wiggler source based on geometrical arguments, regarded as almost self-evident. According to electrodynamics, depth-of-field effects do not exist: we demonstrate this statement both theoretically and numerically, using a well-known first-principle computer code. This fact shows that under the usually accepted approximations, the description of the wiggler brightness turns out to be inconsistent even qualitatively. Therefore, there is a need for a well-defined procedure for computing the brightness from a wiggler source. We accomplish this task based on the use of a Wigner function formalism. We exemplify this formalism in simple limiting cases. We consider the problem of the calculation of the wiggler source size by means of numerical simulations alone, which play the same role of an experiment. We report a significant numerical disagreement between exact calculations and approximations currently used in the literature.

  20. Polyvinylpyrrolidone dewaxing aid for bright stocks

    SciTech Connect

    Achia, B.U.; Shaw, D.H.

    1980-05-20

    Polyvinylpyrrolidone having a number average molecular weight ranging from about 150,000 to 400,000 has been found to be an effective dewaxing aid for bright stock in ketone dewaxing processes. Using as little as 100 ppm based on the waxy oil can result in almost a 50% increase in the filter rate of the dewaxed oils from the wax.

  1. Sky brightness during eclipses: a review.

    PubMed

    Silverman, S M; Mullen, E G

    1975-12-01

    This paper is abstracted from the introductory section of "Sky Brightness During Eclipses: A Compendium from the Literature," AFCRL-TR-74-0363, Special Reports 180, Air Force Cambridge Research Laboratories, Hanscom AFB, Massachusetts 01731. This report should be consulted for fuller details and tables.

  2. Alberta Associations for Bright Children Members' Handbook.

    ERIC Educational Resources Information Center

    Alberta Association for Bright Children, Edmonton.

    This handbook is designed to provide information to parents of gifted children in Alberta, Canada. The handbook outlines the mission and objectives of the Alberta Associations for Bright Children and describes the structure of the non-profit organization. The booklet then addresses: (1) the characteristics of gifted children; (2) the rights of…

  3. Sky brightness during eclipses: a review.

    PubMed

    Silverman, S M; Mullen, E G

    1975-12-01

    This paper is abstracted from the introductory section of "Sky Brightness During Eclipses: A Compendium from the Literature," AFCRL-TR-74-0363, Special Reports 180, Air Force Cambridge Research Laboratories, Hanscom AFB, Massachusetts 01731. This report should be consulted for fuller details and tables. PMID:20155120

  4. Probable Bright Supernovae discovered by PSST

    NASA Astrophysics Data System (ADS)

    Smith, K. W.; Wright, D.; Smartt, S. J.; Huber, M.; Chambers, K. C.; Flewelling, H.; Willman, M.; Primak, N.; Schultz, A.; Gibson, B.; Magnier, E.; Waters, C.; Tonry, J.; Wainscoat, R. J.; Foley, R. J.; Jha, S. W.; Rest, A.; Scolnic, D.

    2016-01-01

    Three bright transients, which are probable supernovae, have been discovered as part of the Pan-STARRS Survey for Transients (PSST). Information on all objects discovered by the Pan-STARRS Survey for Transients is available at http://star.pst.qub.ac.uk/ps1threepi/ (see Huber et al. ATel #7153).

  5. Probable Bright Supernova discovered by PSST

    NASA Astrophysics Data System (ADS)

    Smith, K. W.; Wright, D.; Smartt, S. J.; Young, D. R.; Huber, M.; Chambers, K. C.; Flewelling, H.; Willman, M.; Primak, N.; Schultz, A.; Gibson, B.; Magnier, E.; Waters, C.; Tonry, J.; Wainscoat, R. J.; Foley, R. J.; Jha, S. W.; Rest, A.; Scolnic, D.

    2016-09-01

    A bright transient, which is a probable supernova, has been discovered as part of the Pan-STARRS Survey for Transients (PSST). Information on all objects discovered by the Pan-STARRS Survey for Transients is available at http://star.pst.qub.ac.uk/ps1threepi/ (see Huber et al. ATel #7153).

  6. Simultaneous brightness contrast of foraging Papilio butterflies

    PubMed Central

    Kinoshita, Michiyo; Takahashi, Yuki; Arikawa, Kentaro

    2012-01-01

    This study focuses on the sense of brightness in the foraging Japanese yellow swallowtail butterfly, Papilio xuthus. We presented two red discs of different intensity on a grey background to butterflies, and trained them to select one of the discs. They were successfully trained to select either a high intensity or a low intensity disc. The trained butterflies were tested on their ability to perceive brightness in two different protocols: (i) two orange discs of different intensity presented on the same intensity grey background and (ii) two orange discs of the same intensity separately presented on a grey background that was either higher or lower in intensity than the training background. The butterflies trained to high intensity red selected the orange disc of high intensity in protocol 1, and the disc on the background of low intensity grey in protocol 2. We obtained similar results in another set of experiments with purple discs instead of orange discs. The choices of the butterflies trained to low intensity red were opposite to those just described. Taken together, we conclude that Papilio has the ability to learn brightness and darkness of targets independent of colour, and that they have the so-called simultaneous brightness contrast. PMID:22179808

  7. Bright Meteor Lights Up Atlanta Skies

    NASA Video Gallery

    This video shows a very bright meteor that streaked over the skies of Atlanta, Ga., on the night of Aug. 28, 2011. The view is from an all sky camera in Cartersville, Ga., operated by NASA’s Mars...

  8. REX, a 5-MV pulsed-power source for driving high-brightness electron beam diodes

    SciTech Connect

    Carlson, R.L.; Kauppila, T.J.; Ridlon, R.N.

    1991-01-01

    The Relativistic Electron-beam Experiment, or REX accelerator, is a pulsed-power source capable of driving a 100-ohm load at 5 MV, 50 kA, 45 ns (FWHM) with less than a 10-ns rise and 15-ns fall time. This paper describes the pulsed-power modifications, modelling, and extensive measurements on REX to allow it to drive high impedance (100s of ohms) diode loads with a shaped voltage pulse. A major component of REX is the 1.83-m-diam {times} 25.4-cm-thick Lucite insulator with embedded grading rings that separates the output oil transmission line from the vacuum vessel that contains the re-entrant anode and cathode assemblies. A radially tailored, liquid-based resistor provides a stiff voltage source that is insensitive to small variations of the diode current and, in addition, optimizes the electric field stress across the vacuum side of the insulator. The high-current operation of REX employs both multichannel peaking and point-plane diverter switches. This mode reduces the prepulse to less than 2 kV and the postpulse to less than 5% of the energy delivered to the load. Pulse shaping for the present diode load is done through two L-C transmission line filters and a tapered, glycol-based line adjacent to the water PFL and output switch. This has allowed REX to drive a diode producing a 4-MV, 4.5-kA, 55-ns flat-top electron beam with a normalized Lapostolle emittance of 0.96 mm-rad corresponding to a beam brightness in excess of 4.4 {times} 10{sup 8} A/m{sup 2} {minus}rad{sup 2}. 6 refs., 13 figs.

  9. Resistance Analyses of HCV NS3/4A Protease and NS5B Polymerase from Clinical Studies of Deleobuvir and Faldaprevir

    PubMed Central

    Berger, Kristi L.; Sarrazin, Christoph; Nelson, David R.; Scherer, Joseph; Sha, Nanshi; Marquis, Martin; Côté-Martin, Alexandra; Vinisko, Richard; Stern, Jerry O.; Mensa, Federico J.; Kukolj, George

    2016-01-01

    Background & Aim The resistance profile of anti-hepatitis C virus (HCV) agents used in combination is important to guide optimal treatment regimens. We evaluated baseline and treatment-emergent NS3/4A and NS5B amino-acid variants among HCV genotype (GT)-1a and -1b-infected patients treated with faldaprevir (HCV protease inhibitor), deleobuvir (HCV polymerase non-nucleoside inhibitor), and ribavirin in multiple clinical studies. Methods HCV NS3/4A and NS5B population sequencing (Sanger method) was performed on all baseline plasma samples (n = 1425 NS3; n = 1556 NS5B) and on post-baseline plasma samples from patients with virologic failure (n = 113 GT-1a; n = 221 GT-1b). Persistence and time to loss of resistance-associated variants (RAVs) was estimated using Kaplan–Meier analysis. Results Faldaprevir RAVs (NS3 R155 and D168) and deleobuvir RAVs (NS5B 495 and 496) were rare (<1%) at baseline. Virologic response to faldaprevir/deleobuvir/ribavirin was not compromised by common baseline NS3 polymorphisms (e.g. Q80K in 17.5% of GT-1a) or by NS5B A421V, present in 20% of GT-1a. In GT-1b, alanine at NS5B codon 499 (present in 15% of baseline sequences) was associated with reduced response. Treatment-emergent RAVs consolidated previous findings: NS3 R155 and D168 were key faldaprevir RAVs; NS5B A421 and P495 were key deleobuvir RAVs. Among on-treatment virologic breakthroughs, RAVs emerged in both NS3 and NS5B (>90%). Virologic relapse was associated with RAVs in both NS3 and NS5B (53% GT-1b; 52% GT-1b); some virologic relapses had NS3 RAVs only (47% GT-1a; 17% GT-1b). Median time to loss of GT-1b NS5B P495 RAVs post-treatment (5 months) was less than that of GT-1b NS3 D168 (8.5 months) and GT-1a R155 RAVs (11.5 months). Conclusion Faldaprevir and deleobuvir RAVs are more prevalent among virologic failures than at baseline. Treatment response was not compromised by common NS3 polymorphisms; however, alanine at NS5B amino acid 499 at baseline (wild-type in GT-1a

  10. Crystal structure of Zika virus NS2B-NS3 protease in complex with a boronate inhibitor.

    PubMed

    Lei, Jian; Hansen, Guido; Nitsche, Christoph; Klein, Christian D; Zhang, Linlin; Hilgenfeld, Rolf

    2016-07-29

    The ongoing Zika virus (ZIKV) outbreak is linked to severe neurological disorders. ZIKV relies on its NS2B/NS3 protease for polyprotein processing; hence, this enzyme is an attractive drug target. The 2.7 angstrom; crystal structure of ZIKV protease in complex with a peptidomimetic boronic acid inhibitor reveals a cyclic diester between the boronic acid and glycerol. The P2 4-aminomethylphenylalanine moiety of the inhibitor forms a salt-bridge with the nonconserved Asp(83) of NS2B; ion-pairing between Asp(83) and the P2 residue of the substrate likely accounts for the enzyme's high catalytic efficiency. The unusual dimer of the ZIKV protease:inhibitor complex seen in the crystal may provide a model for assemblies formed at high local concentrations of protease at the endoplasmatic reticulum membrane, the site of polyprotein processing. PMID:27386922

  11. Further theoretical insight into the reaction mechanism of the hepatitis C NS3/NS4A serine protease

    NASA Astrophysics Data System (ADS)

    Martínez-González, José Ángel; Rodríguez, Alex; Puyuelo, María Pilar; González, Miguel; Martínez, Rodrigo

    2015-01-01

    The main reactions of the hepatitis C virus NS3/NS4A serine protease are studied using the second-order Møller-Plesset ab initio method and rather large basis sets to correct the previously reported AM1/CHARMM22 potential energy surfaces. The reaction efficiencies measured for the different substrates are explained in terms of the tetrahedral intermediate formation step (the rate-limiting process). The energies of the barrier and the corresponding intermediate are so close that the possibility of a concerted mechanism is open (especially for the NS5A/5B substrate). This is in contrast to the suggested general reaction mechanism of serine proteases, where a two-step mechanism is postulated.

  12. Crystal structure of Zika virus NS2B-NS3 protease in complex with a boronate inhibitor.

    PubMed

    Lei, Jian; Hansen, Guido; Nitsche, Christoph; Klein, Christian D; Zhang, Linlin; Hilgenfeld, Rolf

    2016-07-29

    The ongoing Zika virus (ZIKV) outbreak is linked to severe neurological disorders. ZIKV relies on its NS2B/NS3 protease for polyprotein processing; hence, this enzyme is an attractive drug target. The 2.7 angstrom; crystal structure of ZIKV protease in complex with a peptidomimetic boronic acid inhibitor reveals a cyclic diester between the boronic acid and glycerol. The P2 4-aminomethylphenylalanine moiety of the inhibitor forms a salt-bridge with the nonconserved Asp(83) of NS2B; ion-pairing between Asp(83) and the P2 residue of the substrate likely accounts for the enzyme's high catalytic efficiency. The unusual dimer of the ZIKV protease:inhibitor complex seen in the crystal may provide a model for assemblies formed at high local concentrations of protease at the endoplasmatic reticulum membrane, the site of polyprotein processing.

  13. Novel fullerene derivatives as dual inhibitors of Hepatitis C virus NS5B polymerase and NS3/4A protease.

    PubMed

    Kataoka, Hiroki; Ohe, Tomoyuki; Takahashi, Kyoko; Nakamura, Shigeo; Mashino, Tadahiko

    2016-10-01

    We evaluated the Hepatitis C virus (HCV) NS5B polymerase and HCV NS3/4A protease inhibition activities of a new set of proline-type fullerene derivatives. All of the compounds had the potential to inhibit both the enzymes, indicating that the fullerene derivatives may be dual inhibitors against NS5B and NS3/4A and could be novel lead compounds for the treatment of HCV infections. PMID:27597249

  14. A straightforward experimental approach to expression, purification, refolding, and enzymatic analysis of recombinant dengue virus NS2B(H)-NS3pro protease.

    PubMed

    Junaid, M; Angsuthanasombat, C; Wikberg, J E S; Ali, N; Katzenmeier, G

    2013-08-01

    Dengue virus threatens around 2.5 billion people worldwide; about 50 million become infected every year, and yet no vaccine or drug is available for prevention and/or treatment. The flaviviral NS2B-NS3pro complex is indispensable for flaviviral replication and is considered to be an important drug target. The aim of this study was to develop a simple and generally applicable experimental strategy to construct, purify, and assay a highly active recombinant NS2B(H)-NS3pro complex that would be useful for high-throughput screening of potential inhibitors. The sequence of NS2B(H)-NS3pro was generated by overlap extension PCR (SOE-PCR) and cloned into the pTrcHisA vector. Hexahistidine-tagged NS2B(H)-NS3pro complex was expressed in E. coli predominantly as insoluble protein and purified to >95% purity by single-step immobilized metal affinity chromatography. SDS-PAGE followed by immunoblotting of the purified enzyme demonstrated the presence of the NS2B(H)-NS3pro precursor and its autocleavage products, NS3pro and NS2B(H), as 37, 21, and 10 kDa bands, respectively. Kinetic parameters, Km, kcat, and kcat/Km for the fluorophore-linked protease model substrate Ac-nKRR-amc were obtained using inner-filter effect correction. The kinetic parameters Km, kcat, and kcat/Km for Ac-nKRR-amc substrate were 100 µM, 0.112 s(-1), and 1120 M(-1)·s(-1), respectively. A simplified procedure for the cloning, overexpression, and purification of the NS2B(H)-NS3pro complex was applied, and a highly active recombinant NS2B(H)-NS3pro complex was obtained that could be useful for the design of high-throughput assays aimed at flaviviral inhibitor discovery.

  15. Mosquito densonucleosis virus non-structural protein NS2 is necessary for a productive infection

    SciTech Connect

    Azarkh, Eugene; Robinson, Erin; Hirunkanokpun, Supanee; Afanasiev, Boris; Kittayapong, Pattamaporn; Carlson, Jonathan Corsini, Joe

    2008-04-25

    Mosquito densonucleosis viruses synthesize two non-structural proteins, NS1 and NS2. While NS1 has been studied relatively well, little is known about NS2. Antiserum was raised against a peptide near the N-terminus of NS2, and used to conduct Western blot analysis and immuno-fluorescence assays. Western blots revealed a prominent band near the expected size (41 kDa). Immuno-fluorescence studies of mosquito cells transfected with AeDNV indicate that NS2 has a wider distribution pattern than does NS1, and the distribution pattern appears to be a function of time post-infection. Nuclear localization of NS2 requires intact C-terminus but does not require additional viral proteins. Mutations ranging from complete NS2 knock-out to a single missense amino acid substitution in NS2 can significantly reduce viral replication and production of viable progeny.

  16. Interaction cloning of NS1-I, a human protein that binds to the nonstructural NS1 proteins of influenza A and B viruses.

    PubMed Central

    Wolff, T; O'Neill, R E; Palese, P

    1996-01-01

    The yeast interaction trap system was used to identify, NS1-I (for NS1 interactor), which is a human protein that binds to the nonstructural NS1 protein of the influenza A virus. NS1-I is a human homolog of the porcine 17beta-estradiol dehydrogenase precursor protein, to which it is 84% identical. We detected only one NS1-I mRNA species, of about 3.0 kb, in HeLa cells, and the NS1-I cDNA was found to have a coding capacity for a 79.6-kDa protein. However, immunoblot analysis detected predominantly a 55-kDa protein in human cells, suggesting that NS1-I, like the porcine 17beta-estradiol dehydrogenase, is posttranslationally processed. Using an in vitro coprecipitation assay, we showed that NS1-I interacts with NS1 proteins from extracts of cells infected with five different influenza A virus strains as well as with the NS1 of an influenza B virus. The fact that influenza A and influenza B virus NS1 proteins bind to NS1-I suggests that this cellular protein plays a role in the influenza virus life cycle. PMID:8764047

  17. [System of ns time-resolved spectroscopy diagnosis and radioprotection].

    PubMed

    Yao, Wei-Bo; Guo, Jian-Ming; Zhang, Yong-min; Tang, Jun-Ping; Cheng, Liang; Xu, Qi-fuo

    2014-06-01

    Cathode plasma of high current electron beam diode is an important research on high power microwave and strong pulsed radio accelerator. It is a reliable method to study cathode plasma by diagnosing the cathode plasma parameters with non-contact spectroscopy measurement system. The present paper introduced the work principle, system composition and performance of the nanosecond (ns) time-resolved spectroscopy diagnosis system. Furthermore, it introduced the implementing method and the temporal relation of lower jitter synchronous trigger system. Simultaneously, the authors designed electromagnetic and radio shield room to protect the diagnosis system due to the high electromagnetic and high X-ray and γ-ray radiation, which seriously interferes with the system. Time-resolved spectroscopy experiment on brass (H62) cathode shows that, the element and matter composition of cathode plasma is clearly increase with the increase in the diode pulsed voltage and current magnitude. The spectroscopy diagnosis system could be of up to 10 ns time resolve capability. It's least is 2 ns. Synchronous trigger system's jitter is less than 4 ns. The spectroscopy diagnosis system will open a new way to study the cathode emission mechanism in depth. PMID:25358142

  18. Circadian Phase-Shifting Effects of Bright Light, Exercise, and Bright Light + Exercise

    PubMed Central

    Kline, Christopher E.; Elliott, Jeffrey A.; Zielinski, Mark R.; Devlin, Tina M.; Moore, Teresa A.

    2016-01-01

    Limited research has compared the circadian phase-shifting effects of bright light and exercise and additive effects of these stimuli. The aim of this study was to compare the phase-delaying effects of late night bright light, late night exercise, and late evening bright light followed by early morning exercise. In a within-subjects, counterbalanced design, 6 young adults completed each of three 2.5-day protocols. Participants followed a 3-h ultra-short sleep-wake cycle, involving wakefulness in dim light for 2h, followed by attempted sleep in darkness for 1 h, repeated throughout each protocol. On night 2 of each protocol, participants received either (1) bright light alone (5,000 lux) from 2210–2340 h, (2) treadmill exercise alone from 2210–2340 h, or (3) bright light (2210–2340 h) followed by exercise from 0410–0540 h. Urine was collected every 90 min. Shifts in the 6-sulphatoxymelatonin (aMT6s) cosine acrophase from baseline to post-treatment were compared between treatments. Analyses revealed a significant additive phase-delaying effect of bright light + exercise (80.8 ± 11.6 [SD] min) compared with exercise alone (47.3 ± 21.6 min), and a similar phase delay following bright light alone (56.6 ± 15.2 min) and exercise alone administered for the same duration and at the same time of night. Thus, the data suggest that late night bright light followed by early morning exercise can have an additive circadian phase-shifting effect. PMID:27103935

  19. TFaNS-Tone Fan Noise Design/Prediction System: Users' Manual TFaNS Version 1.5

    NASA Technical Reports Server (NTRS)

    Topol, David A.; Huff, Dennis L. (Technical Monitor)

    2003-01-01

    TFaNS is the Tone Fan Noise Design/Prediction System developed by Pratt & Whitney under contract to NASA Glenn. The purpose of this system is to predict tone noise emanating from a fan stage including the effects of reflection and transmission by the rotor and stator and by the duct inlet and nozzle. The first version of this design system was developed under a previous NASA contract. Several improvements have been made to TFaNS. This users' manual shows how to run this new system. TFaNS consists of the codes that compute the acoustic properties (reflection and transmission coefficients) of the various elements and writes them to files, CUP3D Fan Noise Coupling Code that reads these files, solves the coupling problem, and outputs the desired noise predictions, and AWAKEN CFD/Measured Wake Postprocessor which reformats CFD wake predictions and/or measured wake data so they can be used by the system. This report provides information on code input and file structure essential for potential users of TFaNS.

  20. Image mosaic with color and brightness correction

    NASA Astrophysics Data System (ADS)

    Zhao, Yili; Xu, Dan; Pan, Zhigeng

    2004-03-01

    Image mosaic is comprised of building a large field of view from a sequence of smaller images. It can be performed by registering, projective warping, resampling and compositing a serials of images. Due to the many possible factors for color and brightness variations when taking images, it is possible to lead to misalignment and obtain poor stitching result. Despite image mosaic can be manually adjusted using some photo editors like PhotoShop, this is not only tedious but also requires skills, knowledge and experience. Automatic adjustment is therefore desirable. By converting images to lαβ space and applying a special statistical analysis, color and brightness correction can be done automatically and improved image mosaic can be obtained.

  1. Brightness increase in an LCD stereo display

    NASA Astrophysics Data System (ADS)

    Rallison, Richard D.; Schicker, Scott R.

    1994-05-01

    A practical head mounted display (HMD) has to be light enough and bright enough to wear and view without undue strain on the users head or eyes. A 10 pound CRT based helmet is not always out of the question but binocular or stereo HMDs using LCDs rather than CRTs need only weigh in at around one pound complete with electronics and are far more comfortable to wear. The space bandwidth product or pixel count of LCDs is now approaching that of CRT type displays but LCDs could use a big boost in brightness, especially for see thru designs. The see thru or head up style has many user advantages and this paper addresses ways to more efficiently transmit photons from the source to the eye in one such design. All of the components that are used to improve performance may be made holographically or in an alternate fashion. The most practical method of construction is probably a toss up for some components.

  2. The Hepatitis C Virus NS5A Stimulates NS5B During In Vitro RNA Synthesis in a Template Specific Manner

    PubMed Central

    Quezada, Elizabeth M; Kane, Caroline M

    2009-01-01

    The hepatitis C virus (HCV) NS5B protein contains the RNA dependent RNA polymerase (RdRp) activity that catalyzes the synthesis of the viral genome with other host and viral factors. NS5A is an HCV-encoded protein previously shown to localize to the replisome and be necessary for viral replication. However, its role in replication has not been defined. Using an in vitro biochemical assay, we detected a stimulatory effect of NS5A on the NS5B replication reaction with minimal natural templates. NS5A stimulates replication by NS5B on two templates derived from the 3’ end of the RNA genome (4 fold ± 1.3 fold). A pre-incubation step with the two proteins prior to the replication reaction and substoichiometric levels of NS5A are required for detecting stimulation. With a template derived from the 3’end complementary to the RNA genome (the negative strand) no stimulation was observed. Furthermore, with a synthetic template that allows studying different phases of replication, NS5A stimulates NS5B during elongation. These findings suggest that NS5A stimulates NS5B during synthesis of the complementary (i.e., negative) strand of the RNA genome. PMID:19590581

  3. N-bright-bright and N-dark-dark solitons of the coupled generalized nonlinear Schrödinger equations

    NASA Astrophysics Data System (ADS)

    Vishnu Priya, N.; Senthilvelan, M.

    2016-07-01

    We construct N-bright-bright and N-dark-dark soliton solutions of an integrable two coupled generalized nonlinear Schrödinger (CGNLS) equation for arbitrary values of system parameters. These solutions are more general than the reported one. While the bright-bright solitons are captured in the focusing regime of CGNLS equation, the dark-dark soliton solutions are identified in the defocusing regime. We present N-bright-bright solitons in the Gram determinant forms and prove that these determinant forms satisfy the Hirota bilinear equations.

  4. Australia 31-GHz brightness temperature exceedance statistics

    NASA Technical Reports Server (NTRS)

    Gary, B. L.

    1988-01-01

    Water vapor radiometer measurements were made at DSS 43 during an 18 month period. Brightness temperatures at 31 GHz were subjected to a statistical analysis which included correction for the effects of occasional water on the radiometer radome. An exceedance plot was constructed, and the 1 percent exceedance statistics occurs at 120 K. The 5 percent exceedance statistics occurs at 70 K, compared with 75 K in Spain. These values are valid for all of the three month groupings that were studied.

  5. Nonlinear Brightness Optimization in Compton Scattering

    DOE PAGESBeta

    Hartemann, Fred V.; Wu, Sheldon S. Q.

    2013-07-26

    In Compton scattering light sources, a laser pulse is scattered by a relativistic electron beam to generate tunable x and gamma rays. Because of the inhomogeneous nature of the incident radiation, the relativistic Lorentz boost of the electrons is modulated by the ponderomotive force during the interaction, leading to intrinsic spectral broadening and brightness limitations. We discuss these effects, along with an optimization strategy to properly balance the laser bandwidth, diffraction, and nonlinear ponderomotive force.

  6. UV-bright stars in globular clusters

    NASA Technical Reports Server (NTRS)

    Landsman, Wayne B.

    1994-01-01

    This paper highlights globular cluster studies with Ultraviolet Imaging Telescope (UIT) in three areas: the discrepancy between observed ultraviolet HB magnitudes and predictions of theoretical HB models; the discovery of two hot subdwarfs in NGC 1851, a globular not previously known to contain such stars; and spectroscopic follow up of newly identified UV-bright stars in M79 and w Cen. I also present results of a recent observation of NGC 6397 with the Voyager ultraviolet spectrometer.

  7. Extreme Ultraviolet Explorer Bright Source List

    NASA Technical Reports Server (NTRS)

    Malina, Roger F.; Marshall, Herman L.; Antia, Behram; Christian, Carol A.; Dobson, Carl A.; Finley, David S.; Fruscione, Antonella; Girouard, Forrest R.; Hawkins, Isabel; Jelinsky, Patrick

    1994-01-01

    Initial results from the analysis of the Extreme Ultraviolet Explorer (EUVE) all-sky survey (58-740 A) and deep survey (67-364 A) are presented through the EUVE Bright Source List (BSL). The BSL contains 356 confirmed extreme ultraviolet (EUV) point sources with supporting information, including positions, observed EUV count rates, and the identification of possible optical counterparts. One-hundred twenty-six sources have been detected longward of 200 A.

  8. Low-Dispersion Observations of Bright Stars

    NASA Astrophysics Data System (ADS)

    Fitzpatrick, Edward L.

    Seven US2 observing shifts are being requested to obtain low-dispersion SWP and LWP spectra of approximately 15 bright, nearby early-type stars. The targets are taken from the 10-year old effective temperature and bolometric correction study of Code, Davis, Bless, and Hanbury Brown (CDBB). The CDBB stars represent the only sample of stars for which angular diameter measurements are available. The stars which we plan to observe have been unobservable with the low-dispersion mode of IUE in the past because of their extreme brightness; however, the recent refinements in the fast-trailing technique now allow optimally exposed spectra to be obtained. With the new spectra and with Archival spectra which are available for some of the less bright CDBB stars, we plan to repeat the earlier effective temperature and bolometric correction determinations, taking advantage of the higher photometric stability and higher resolution of IUE over previous ultraviolet missions and utilizing improvements in the ground-based optical/lR data and calibrations. This study will tie the large IUE database into a system of fundamental stellar effective temperature and bolometric correction determinations.

  9. Personal audio with a planar bright zone.

    PubMed

    Coleman, Philip; Jackson, Philip J B; Olik, Marek; Pedersen, Jan Abildgaard

    2014-10-01

    Reproduction of multiple sound zones, in which personal audio programs may be consumed without the need for headphones, is an active topic in acoustical signal processing. Many approaches to sound zone reproduction do not consider control of the bright zone phase, which may lead to self-cancellation problems if the loudspeakers surround the zones. Conversely, control of the phase in a least-squares sense comes at a cost of decreased level difference between the zones and frequency range of cancellation. Single-zone approaches have considered plane wave reproduction by focusing the sound energy in to a point in the wavenumber domain. In this article, a planar bright zone is reproduced via planarity control, which constrains the bright zone energy to impinge from a narrow range of angles via projection in to a spatial domain. Simulation results using a circular array surrounding two zones show the method to produce superior contrast to the least-squares approach, and superior planarity to the contrast maximization approach. Practical performance measurements obtained in an acoustically treated room verify the conclusions drawn under free-field conditions. PMID:25324075

  10. Bright Ray Craters in Ganymede's Northern Hemisphere

    NASA Technical Reports Server (NTRS)

    1979-01-01

    GANYMEDE COLOR PHOTOS: This color picture as acquired by Voyager 1 during its approach to Ganymede on Monday afternoon (the 5th of March). At ranges between about 230 to 250 thousand km. The images show detail on the surface with a resolution of four and a half km. This picture is of a region in the northern hemisphere near the terminator. It shows a variety of impact structures, including both razed and unrazed craters, and the odd, groove-like structures discovered by Voyager in the lighter regions. The most striking features are the bright ray craters which have a distinctly 'bluer' color appearing white against the redder background. Ganymede's surface is known to contain large amounts of surface ice and it appears that these relatively young craters have spread bright fresh ice materials over the surface. Likewise, the lighter color and reflectivity of the grooved areas suggests that here, too, there is cleaner ice. We see ray craters with all sizes of ray patterns, ranging from extensive systems of the crater in the southern part of this picture, which has rays at least 300-500 kilometers long, down to craters which have only faint remnants of bright ejects patterns (such as several of the craters in the southern half of PIA01516; P21262). This variation suggests that, as on the Moon, there are processes which act to darken ray material, probably 'gardening' by micrometeoroid impact. JPL manages and controls the Voyager project for NASA's Office of Space Science.

  11. Search for bright stars with infrared excess

    SciTech Connect

    Raharto, Moedji

    2014-03-24

    Bright stars, stars with visual magnitude smaller than 6.5, can be studied using small telescope. In general, if stars are assumed as black body radiator, then the color in infrared (IR) region is usually equal to zero. Infrared data from IRAS observations at 12 and 25μm (micron) with good flux quality are used to search for bright stars (from Bright Stars Catalogues) with infrared excess. In magnitude scale, stars with IR excess is defined as stars with IR color m{sub 12}−m{sub 25}>0; where m{sub 12}−m{sub 25} = −2.5log(F{sub 12}/F{sub 25})+1.56, where F{sub 12} and F{sub 25} are flux density in Jansky at 12 and 25μm, respectively. Stars with similar spectral type are expected to have similar color. The existence of infrared excess in the same spectral type indicates the existence of circum-stellar dust, the origin of which is probably due to the remnant of pre main-sequence evolution during star formation or post AGB evolution or due to physical process such as the rotation of those stars.

  12. Bright light activates a trigeminal nociceptive pathway

    PubMed Central

    Okamoto, Keiichiro; Tashiro, Akimasa; Chang, Zheng; Bereiter, David A.

    2010-01-01

    Bright light can cause ocular discomfort and/or pain; however, the mechanism linking luminance to trigeminal nerve activity is not known. In this study we identify a novel reflex circuit necessary for bright light to excite nociceptive neurons in superficial laminae of trigeminal subnucleus caudalis (Vc/C1). Vc/C1 neurons encoded light intensity and displayed a long delay (>10 s) for activation. Microinjection of lidocaine into the eye or trigeminal root ganglion (TRG) inhibited light responses completely, whereas topical application onto the ocular surface had no effect. These findings indicated that light-evoked Vc/C1 activity was mediated by an intraocular mechanism and transmission through the TRG. Disrupting local vasomotor activity by intraocular microinjection of the vasoconstrictive agents, norepinephrine or phenylephrine, blocked light-evoked neural activity, whereas ocular surface or intra-TRG microinjection of norepinephrine had no effect. Pupillary muscle activity did not contribute since light-evoked responses were not altered by atropine. Microinjection of lidocaine into the superior salivatory nucleus diminished light-evoked Vc/C1 activity and lacrimation suggesting that increased parasympathetic outflow was critical for light-evoked responses. The reflex circuit also required input through accessory visual pathways since both Vc/C1 activity and lacrimation were prevented by local blockade of the olivary pretectal nucleus. These findings support the hypothesis that bright light activates trigeminal nerve activity through an intraocular mechanism driven by a luminance-responsive circuit and increased parasympathetic outflow to the eye. PMID:20206444

  13. Bright and Dark Slopes on Ganymede

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Ridges on the edge of Ganymede's north polar cap show bright east-facing slopes and dark west-facing slopes with troughs of darker material below the larger ridges. North is to the top. The bright slopes may be due to grain size differences, differences in composition between the original surface and the underlying material, frost deposition, or illumination effects. The large 2.4 kilometer (1.5 mile) diameter crater in this image shows frost deposits located on the north-facing rim slope, away from the sun. A smaller 675 meter (2200 foot) diameter crater in the center of the image is surrounded by a bright deposit which may be ejecta from the impact. Ejecta deposits such as this are uncommon for small craters on Ganymede. This image measures 18 by 19 kilometers (11 by 12 miles) and has a resolution of 45 meters (148 feet) per pixel. NASA's Galileo spacecraft obtained this image on September 6, 1996 during its second orbit around Jupiter.

    The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology (Caltech).

    This image and other images and data received from Galileo are posted on the World Wide Web, on the Galileo mission home page at URL http://galileo.jpl.nasa.gov. Background information and educational context for the images can be found at URL http://www.jpl.nasa.gov/galileo/sepo

  14. Brightness Changes in Sun-like Stars

    NASA Technical Reports Server (NTRS)

    Henry, Stephen M.; Henry, Gregory W.

    1998-01-01

    Does the Sun's energy output vary with time? Are observable climatic changes on the earth caused by changes in the Sun? Can we gain greater insight into this relation-ship by studying other stars with properties similar to the Sun's? In recent years, satellite observations have shown that the solar irradiance varies in phase with the 1 l-year sunspot cycle. The Sun is brighter by about O.l% at the peak of the sunspot cycle when solar magnetic activity is at its maximum. Over longer intervals, changes in the cart h's climate and solar magnetic activity seem to be correlated. We are using automatic photoelectric telescopes to measure brightness changes in a sample of 150 Sun-like stars. Lowell Observatory astronomers have also observed about 30 of these same stars with a manual telescope in a program that began 10 years before ours. Since these two data sets were acquired with different instruments and so have significant systematic differences, we developed software to combine them accurately and, therefore, extend our observational time coverage. We show sample results of brightness variations over 14 years in several Sun-like stars with different ages. Longitudinal studies like these, combined with cross-sectional studies of the larger sample of stars, may eventually allow us to infer with confidence the Sun's long-term brightness history and its impact on the earth's climate.

  15. Use of parallel validation high-throughput screens to reduce false positives and identify novel dengue NS2B-NS3 protease inhibitors†

    PubMed Central

    Tomlinson, Suzanne M.; Watowich, Stanley J.

    2011-01-01

    Dengue virus (DENV), a mosquito-borne member of the family Flaviviridae, is a significant global pathogen affecting primarily tropical and subtropical regions of the world and placing tremendous burden on the limited medical infrastructure that exists in many of the developing countries located within these regions. Recent outbreaks in developed countries, including Australia (Hanna et al., 2009), France (Laruche et al., 2010), Taiwan (Kuan et al., 2010), and the USA (CDC, 2010), lead many researchers to believe that continued emergence into more temperate latitudes is likely. A primary concern is that there are no approved vaccines or antiviral therapies to treat DENV infections. Since the viral NS2B-NS3 protease (DENV NS2B-NS3pro) is required for virus replication, it provides a strategic target for the development of antiviral drugs. In this study, proof-of-concept high-throughput screenings (HTSs) were performed to unambiguously identify dengue 2 virus (DEN2V) NS2B-NS3pro inhibitors from a library of 2000 compounds. Validation screens were performed in parallel to concurrently eliminate insoluble, auto-fluorescing, and/or nonspecific inhibitors. Kinetic analyses of the hits revealed that parallel substrate fluorophore (AMC) interference controls and trypsin inhibition controls were able to reduce false positive rates due to solubility and fluorophore interference while the trypsin inhibition control additionally eliminated non-specific inhibitors. We identified five DEN2V NS2B-NS3pro inhibitors that also inhibited the related West Nile virus (WNV) protease (NS2B-NS3pro), but did not inhibit the trypsin protease. Biochemical analyses revealed various mechanisms of inhibition including competitive and mixed noncompetitive inhibition, with the lowest Ki values being 12 ± 1.5 μM for DEN2V NS2B-NS3pro and 2 ± 0.2 μM for WNV NS2B-NS3pro. PMID:22193283

  16. Phylogenetic analysis of the NS5 gene of Zika virus.

    PubMed

    Adiga, Rama

    2016-10-01

    ZIKV infection has become a global threat spreading across 31 countries in Central America, South America, and the Caribbean. However, little information is available about the molecular epidemiology of ZIKV. Shared mutation of a threonine residue to alanine at the same position in the C terminal of NS5 sequences was observed in sequences from Colombia, Mexico, Panama, and Martinique. The sequences in the phylogenetic tree fell within the same cluster. Based on shared mutation the presence of a Latin American genotype was proposed. Comparison of African and Asian lineages yielded R29N, N273S, H383Q, and P391S mutation. The study highlights that mutation of amino acids at NS5 may contribute to neutropism of ZIKV. J. Med. Virol. 88:1821-1826, 2016. © 2016 Wiley Periodicals, Inc.

  17. Phylogenetic analysis of the NS5 gene of Zika virus.

    PubMed

    Adiga, Rama

    2016-10-01

    ZIKV infection has become a global threat spreading across 31 countries in Central America, South America, and the Caribbean. However, little information is available about the molecular epidemiology of ZIKV. Shared mutation of a threonine residue to alanine at the same position in the C terminal of NS5 sequences was observed in sequences from Colombia, Mexico, Panama, and Martinique. The sequences in the phylogenetic tree fell within the same cluster. Based on shared mutation the presence of a Latin American genotype was proposed. Comparison of African and Asian lineages yielded R29N, N273S, H383Q, and P391S mutation. The study highlights that mutation of amino acids at NS5 may contribute to neutropism of ZIKV. J. Med. Virol. 88:1821-1826, 2016. © 2016 Wiley Periodicals, Inc. PMID:27335310

  18. NS&T Managment Observations - 1st Quarter

    SciTech Connect

    David Gianotto

    2014-06-01

    The INL Management Observation Program (MOP) is designed to improve managers and supervisors understanding of work being performed by employees and the barriers impacting their success. The MOP also increases workers understanding of managements’ expectations as they relate to safety, security, quality, and work performance. Management observations (observations) are designed to improve the relationship and trust between employees and managers through increased engagement and interactions between managers and researchers in the field. As part of continuous improvement, NS&T management took initiative to focus on the participation and quality of observations in FY 14. This quarterly report is intended to (a) summarize the participation and quality of management’s observations, (b) assess observations for commonalities or trends related to facility or process barriers impacting research, and (c) provide feedback and make recommendations for improvements NS&T’s MOP.

  19. NS&T Management Observations: Quarterly Performance Analysis

    SciTech Connect

    Gianotto, David

    2014-09-01

    The INL Management Observation Program (MOP) is designed to improve managers and supervisors understanding of work being performed by employees and the barriers impacting their success. The MOP also increases workers understanding of managements’ expectations as they relate to safety, security, quality, and work performance. Management observations (observations) are designed to improve the relationship and trust between employees and managers through increased engagement and interactions between managers and researchers in the field. As part of continuous improvement, NS&T management took initiative to focus on the participation and quality of observations in FY-14. This quarterly report is intended to (a) summarize the participation and quality of management’s observations, (b) assess observations for commonalities or trends related to facility or process barriers impacting research, and (c) provide feedback and make recommendations for improvements NS&T’s MOP.

  20. NS&T Management Observations - 3rd Quarter

    SciTech Connect

    David Gianotto

    2014-07-01

    The INL Management Observation Program (MOP) is designed to improve managers and supervisors understanding of work being performed by employees and the barriers impacting their success. The MOP also increases workers understanding of managements’ expectations as they relate to safety, security, quality, and work performance. Management observations are designed to improve the relationship and trust between employees and managers through increased engagement and interactions between managers and researchers in the field. As part of continuous improvement, NS&T management took initiative to focus on the participation and quality of observations in FY 14. This quarterly report is intended to (a) summarize the participation and quality of management’s observations, (b) assess observations for commonalities or trends related to facility or process barriers impacting research, and (c) provide feedback and make recommendations for improvements NS&T’s MOP.

  1. SAT: a Late NS Protein of Porcine Parvovirus

    PubMed Central

    Zádori, Zoltán; Szelei, József; Tijssen, Peter

    2005-01-01

    The genomes of all members of the Parvovirus genus were found to contain a small open reading frame (ORF), designated SAT, with a start codon four or seven nucleotides downstream of the VP2 initiation codon. Green fluorescent protein or FLAG fusion constructs of SAT demonstrated that these ORFs were expressed. Although the SAT proteins of the different parvoviruses are not particularly conserved, they were all predicted to contain a membrane-spanning helix, and mutations in this hydrophobic stretch affected the localization of the SAT protein. SAT colocalized with calreticulin in the membranes of the endoplasmic reticulum and the nucleus. A knockout mutant (SAT−), with an unmodified VP sequence, showed a “slow-spreading” phenotype. These knockout mutants could be complemented with VP2− SAT+ mutant. The SAT protein is a late nonstructural (NS) protein, in contrast to previously identified NS proteins, since it is expressed from the same mRNA as VP2. PMID:16189014

  2. [Isolation of Achromobacter xylosoxidans NS12 and degradation of nitrophenols].

    PubMed

    Wan, Nian-Sheng; Gu, Ji-Dong; Huang, Jin-Hui; Gao, Chuan-De

    2007-02-01

    A nitrophenols-degrading bacterium, strain NS12, was isolated from a mangrove sediment by enrichment culture under aerobic conditions. Based on the analysis of 16S rDNA gene sequence the isolate was identified as Achromobacter xylosoxidans Strain NS12 was able to metabolize both o-nitrophenol (ONP) and p-nitrophenol (PNP) as the sole source of carbon, energy and nitrogen. However, this strain was not able to use 3-nitrophenol (MNP) as the only source of carbon energy and nitrogen for growth. The study demonstrated that when PNP and ONP occurred as a mixed substrate PNP degradation restrained the degradation of ONP and caused the major carbon source shift from ONP to PNP. Moreover, the results showed nitrophenols could be degraded by the indigenous bacteria in mangrove sediment.

  3. 7 CFR 51.2000 - Clean and bright.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 2 2013-01-01 2013-01-01 false Clean and bright. 51.2000 Section 51.2000 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing....2000 Clean and bright. Clean and bright means that the individual filbert and the lot as a whole...

  4. 7 CFR 51.2000 - Clean and bright.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 Agriculture 2 2014-01-01 2014-01-01 false Clean and bright. 51.2000 Section 51.2000 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing....2000 Clean and bright. Clean and bright means that the individual filbert and the lot as a whole...

  5. Discovery of an irreversible HCV NS5B polymerase inhibitor.

    PubMed

    Zeng, Qingbei; Nair, Anilkumar G; Rosenblum, Stuart B; Huang, Hsueh-Cheng; Lesburg, Charles A; Jiang, Yueheng; Selyutin, Oleg; Chan, Tin-Yau; Bennett, Frank; Chen, Kevin X; Venkatraman, Srikanth; Sannigrahi, Mousumi; Velazquez, Francisco; Duca, Jose S; Gavalas, Stephen; Huang, Yuhua; Pu, Haiyan; Wang, Li; Pinto, Patrick; Vibulbhan, Bancha; Agrawal, Sony; Ferrari, Eric; Jiang, Chuan-kui; Li, Cheng; Hesk, David; Gesell, Jennifer; Sorota, Steve; Shih, Neng-Yang; Njoroge, F George; Kozlowski, Joseph A

    2013-12-15

    The discovery of lead compound 2e was described. Its covalent binding to HCV NS5B polymerase enzyme was investigated by X-ray analysis. The results of distribution, metabolism and pharmacokinetics were reported. Compound 2e was demonstrated to be potent (replicon GT-1b EC50 = 0.003 μM), highly selective, and safe in in vitro and in vivo assays.

  6. Membrane interacting regions of Dengue virus NS2A protein.

    PubMed

    Nemésio, Henrique; Villalaín, José

    2014-08-28

    The Dengue virus (DENV) NS2A protein, essential for viral replication, is a poorly characterized membrane protein. NS2A displays both protein/protein and membrane/protein interactions, yet neither its functions in the viral cycle nor its active regions are known with certainty. To highlight the different membrane-active regions of NS2A, we characterized the effects of peptides derived from a peptide library encompassing this protein's full length on different membranes by measuring their membrane leakage induction and modulation of lipid phase behavior. Following this initial screening, one region, peptide dens25, had interesting effects on membranes; therefore, we sought to thoroughly characterize this region's interaction with membranes. This peptide presents an interfacial/hydrophobic pattern characteristic of a membrane-proximal segment. We show that dens25 strongly interacts with membranes that contain a large proportion of lipid molecules with a formal negative charge, and that this effect has a major electrostatic contribution. Considering its membrane modulating capabilities, this region might be involved in membrane rearrangements and thus be important for the viral cycle.

  7. Membrane Interacting Regions of Dengue Virus NS2A Protein

    PubMed Central

    2015-01-01

    The Dengue virus (DENV) NS2A protein, essential for viral replication, is a poorly characterized membrane protein. NS2A displays both protein/protein and membrane/protein interactions, yet neither its functions in the viral cycle nor its active regions are known with certainty. To highlight the different membrane-active regions of NS2A, we characterized the effects of peptides derived from a peptide library encompassing this protein’s full length on different membranes by measuring their membrane leakage induction and modulation of lipid phase behavior. Following this initial screening, one region, peptide dens25, had interesting effects on membranes; therefore, we sought to thoroughly characterize this region’s interaction with membranes. This peptide presents an interfacial/hydrophobic pattern characteristic of a membrane-proximal segment. We show that dens25 strongly interacts with membranes that contain a large proportion of lipid molecules with a formal negative charge, and that this effect has a major electrostatic contribution. Considering its membrane modulating capabilities, this region might be involved in membrane rearrangements and thus be important for the viral cycle. PMID:25119664

  8. Effects of influenza A virus NS1 protein on protein expression: the NS1 protein enhances translation and is not required for shutoff of host protein synthesis.

    PubMed

    Salvatore, Mirella; Basler, Christopher F; Parisien, Jean-Patrick; Horvath, Curt M; Bourmakina, Svetlana; Zheng, Hongyong; Muster, Thomas; Palese, Peter; García-Sastre, Adolfo

    2002-02-01

    The influenza A virus NS1 protein, a virus-encoded alpha/beta interferon (IFN-alpha/beta) antagonist, appears to be a key regulator of protein expression in infected cells. We now show that NS1 protein expression results in enhancement of reporter gene activity from transfected plasmids. This effect appears to be mediated at the translational level, and it is reminiscent of the activity of the adenoviral virus-associated I (VAI) RNA, a known inhibitor of the antiviral, IFN-induced, PKR protein. To study the effects of the NS1 protein on viral and cellular protein synthesis during influenza A virus infection, we used recombinant influenza viruses lacking the NS1 gene (delNS1) or expressing truncated NS1 proteins. Our results demonstrate that the NS1 protein is required for efficient viral protein synthesis in COS-7 cells. This activity maps to the amino-terminal domain of the NS1 protein, since cells infected with wild-type virus or with a mutant virus expressing a truncated NS1 protein-lacking approximately half of its carboxy-terminal end-showed similar kinetics of viral and cellular protein expression. Interestingly, no major differences in host cell protein synthesis shutoff or in viral protein expression were found among NS1 mutant viruses in Vero cells. Thus, another viral component(s) different from the NS1 protein is responsible for the inhibition of host protein synthesis during viral infection. In contrast to the earlier proposal suggesting that the NS1 protein regulates the levels of spliced M2 mRNA, no effects on M2 protein accumulation were seen in Vero cells infected with delNS1 virus.

  9. Detection of dengue NS1 and NS3 proteins in placenta and umbilical cord in fetal and maternal death.

    PubMed

    Nunes, Priscila Conrado Guerra; Paes, Marciano Viana; de Oliveira, Carlos Alberto Basilio; Soares, Ana Carla Gomes; de Filippis, Ana Maria Bispo; Lima, Monique da Rocha Queiroz; de Barcelos Alves, Ada Maria; da Silva, Juliana Fernandes Amorim; de Oliveira Coelho, Janice Mery Chicarino; de Carvalho Rodrigues, Francisco das Chagas; Nogueira, Rita Maria Ribeiro; Dos Santos, Flávia Barreto

    2016-08-01

    In Brazil, dengue is a public health problem with the occurrence of explosive epidemics. This study reports maternal and fetal deaths due to dengue and which tissues of placenta and umbilical cord were analyzed by molecular methods and immunohistochemistry. The dengue NS3 and NS1 detection revealed the viral presence in different cells from placenta and umbilical cord. In the latter, DENV-2 was detected at a viral titer of 1,02 × 10(4) amounts of viral RNA. It was shown that the DENV markers analyzed here may be an alternative approach for dengue fatal cases investigation, especially involving maternal and fetal death. J. Med. Virol. 88:1448-1452, 2016. © 2016 Wiley Periodicals, Inc.

  10. Dark and Bright Ridges on Europa

    NASA Technical Reports Server (NTRS)

    1998-01-01

    This high-resolution image of Jupiter's moon Europa, taken by NASA's Galileo spacecraft camera, shows dark, relatively smooth region at the lower right hand corner of the image which may be a place where warm ice has welled up from below. The region is approximately 30 square kilometers in area. An isolated bright hill stands within it. The image also shows two prominent ridges which have different characteristics; youngest ridge runs from left to top right and is about 5 kilometers in width (about 3.1 miles). The ridge has two bright, raised rims and a central valley. The rims of the ridge are rough in texture. The inner and outer walls show bright and dark debris streaming downslope, some of it forming broad fans. This ridge overlies and therefore must be younger than a second ridge running from top to bottom on the left side of the image. This dark 2 km wide ridge is relatively flat, and has smaller-scale ridges and troughs along its length.

    North is to the top of the picture, and the sun illuminates the surface from the upper left. This image, centered at approximately 14 degrees south latitude and 194 degrees west longitude, covers an area approximately 15 kilometers by 20 kilometers (9 miles by 12 miles). The resolution is 26 meters (85 feet) per picture element. This image was taken on December 16, 1997 at a range of 1300 kilometers (800 miles) by Galileo's solid state imaging system.

    The Jet Propulsion Laboratory, Pasadena, CA manages the Galileo mission for NASA's Office of Space Science, Washington, DC. JPL is an operating division of California Institute of Technology (Caltech).

    This image and other images and data received from Galileo are posted on the World Wide Web, on the Galileo mission home page at URL http://www.jpl.nasa.gov/ galileo.

  11. High brightness LED in confocal microscopy

    NASA Astrophysics Data System (ADS)

    Vakili, Ali; Xiong, Daxi; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2015-03-01

    We have introduced a novel illumination system for line scanning confocal microscopy. Confocal microscopy is a popular imaging tool in many applications specifically in medical imaging. Line scanning confocal microscopes have been proven to provide images with resolution comparable to point scanning microscopes. In the point scanning microscopes, the light is focused onto a diffraction limited spot. A pinhole is placed conjugate to the diffraction limited spot, in front of the detector to reject the light coming from out-of-focus planes. Therefore, confocal microscopy can provide optical sectioning. The size of the pinhole determines the amount of light that reaches the detector. A large pinhole results in a blurry image since more of the out-of-focus light contribute to the image. On the other hand, a smaller pinhole rejects more of the light, leading to a lower signal-to-noise ratio. Ideally it is desired to deliver a larger amount of optical power to the diffraction limited spot to increase the signal-to-noise ratio and have a smaller pinhole to reject more of the out-of-focus light. This is the property of the illumination system. In order to get a good signal-to noise ratio in the image, the light source has to provide sufficient radiance. We have introduced a new illumination system utilizing a high brightness LED in the line scanning confocal microscope. High brightness LEDs provide more optical power compared to ordinary LEDs from a smaller area; they have higher radiance. Preliminary results from our line scanning confocal microscope show that the high brightness LED is able to provide enough radiance to obtain an image with resolution comparable with the same microscope utilizing the laser diode. However, in high frame-rate application higher radiance or lower-noise detection system is required.

  12. Chaos may make black holes bright

    NASA Astrophysics Data System (ADS)

    Levin, Janna

    1999-09-01

    Black holes cannot be seen directly since they absorb light and emit none, the very quality which earned them their name. We suggest that black holes may be seen indirectly through a chaotic defocusing of light. A black hole can capture light from a luminous companion in chaotic orbits before scattering the light in random directions. To a distant observer, the black hole would appear to light up. If the companion were a bright radio pulsar, this estimate suggests the black hole echo could be detectable.

  13. Bright solitary waves in malignant gliomas

    NASA Astrophysics Data System (ADS)

    Pérez-García, Víctor M.; Calvo, Gabriel F.; Belmonte-Beitia, Juan; Diego, David; Pérez-Romasanta, Luis

    2011-08-01

    We put forward a nonlinear wave model describing the fundamental dynamical features of an aggressive type of brain tumors. Our model accounts for the invasion of normal tissue by a proliferating and propagating rim of active glioma cancer cells in the tumor boundary and the subsequent formation of a necrotic core. By resorting to numerical simulations, phase space analysis, and exact solutions we prove that bright solitary tumor waves develop in such systems. Possible implications of our model as a tool to extract relevant patient specific tumor parameters in combination with standard preoperative clinical imaging are also discussed.

  14. Two-color bright squeezed vacuum

    SciTech Connect

    Agafonov, Ivan N.; Chekhova, Maria V.

    2010-07-15

    In a strongly pumped nondegenerate traveling-wave optical parametric amplifier, we produce a two-color squeezed vacuum with up to millions of photons per pulse. Our approach to registering this macroscopic quantum state is direct detection of a large number of transverse and longitudinal modes, which is achieved by making the detection time and area much larger than the coherence time and area, respectively. Using this approach, we obtain a record value of twin-beam squeezing for direct detection of bright squeezed vacuum. This makes direct detection of macroscopic squeezed vacuum a practical tool for quantum information applications.

  15. Raman beam combining for laser brightness enhancement

    DOEpatents

    Dawson, Jay W.; Allen, Graham S.; Pax, Paul H.; Heebner, John E.; Sridharan, Arun K.; Rubenchik, Alexander M.; Barty, Chrisopher B. J.

    2015-10-27

    An optical source capable of enhanced scaling of pulse energy and brightness utilizes an ensemble of single-aperture fiber lasers as pump sources, with each such fiber laser operating at acceptable pulse energy levels. Beam combining involves stimulated Raman scattering using a Stokes' shifted seed beam, the latter of which is optimized in terms of its temporal and spectral properties. Beams from fiber lasers can thus be combined to attain pulses with peak energies in excess of the fiber laser self-focusing limit of 4 MW while retaining the advantages of a fiber laser system of high average power with good beam quality.

  16. Human Bocavirus NS1 and NS1-70 Proteins Inhibit TNF-α-Mediated Activation of NF-κB by targeting p65

    PubMed Central

    Liu, Qingshi; Zhang, Zhenfeng; Zheng, Zhenhua; Zheng, Caishang; Liu, Yan; Hu, Qinxue; Ke, Xianliang; Wang, Hanzhong

    2016-01-01

    Human bocavirus (HBoV), a parvovirus, is a single-stranded DNA etiologic agent causing lower respiratory tract infections in young children worldwide. Nuclear factor kappa B (NF-κB) transcription factors play crucial roles in clearance of invading viruses through activation of many physiological processes. Previous investigation showed that HBoV infection could significantly upregulate the level of TNF-α which is a strong NF-κB stimulator. Here we investigated whether HBoV proteins modulate TNF-α–mediated activation of the NF-κB signaling pathway. We showed that HBoV NS1 and NS1-70 proteins blocked NF-κB activation in response to TNF-α. Overexpression of TNF receptor-associated factor 2 (TRAF2)-, IκB kinase alpha (IKKα)-, IκB kinase beta (IKKβ)-, constitutively active mutant of IKKβ (IKKβ SS/EE)-, or p65-induced NF-κB activation was inhibited by NS1 and NS1-70. Furthermore, NS1 and NS1-70 didn’t interfere with TNF-α-mediated IκBα phosphorylation and degradation, nor p65 nuclear translocation. Coimmunoprecipitation assays confirmed the interaction of both NS1 and NS1-70 with p65. Of note, NS1 but not NS1-70 inhibited TNF-α-mediated p65 phosphorylation at ser536. Our findings together indicate that HBoV NS1 and NS1-70 inhibit NF-κB activation. This is the first time that HBoV has been shown to inhibit NF-κB activation, revealing a potential immune-evasion mechanism that is likely important for HBoV pathogenesis. PMID:27329558

  17. Coronal Bright Points Associated with Minifilament Eruptions

    NASA Astrophysics Data System (ADS)

    Hong, Junchao; Jiang, Yunchun; Yang, Jiayan; Bi, Yi; Li, Haidong; Yang, Bo; Yang, Dan

    2014-12-01

    Coronal bright points (CBPs) are small-scale, long-lived coronal brightenings that always correspond to photospheric network magnetic features of opposite polarity. In this paper, we subjectively adopt 30 CBPs in a coronal hole to study their eruptive behavior using data from the Atmospheric Imaging Assembly (AIA) and the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory. About one-quarter to one-third of the CBPs in the coronal hole go through one or more minifilament eruption(s) (MFE(s)) throughout their lifetimes. The MFEs occur in temporal association with the brightness maxima of CBPs and possibly result from the convergence and cancellation of underlying magnetic dipoles. Two examples of CBPs with MFEs are analyzed in detail, where minifilaments appear as dark features of a cool channel that divide the CBPs along the neutral lines of the dipoles beneath. The MFEs show the typical rising movements of filaments and mass ejections with brightenings at CBPs, similar to large-scale filament eruptions. Via differential emission measure analysis, it is found that CBPs are heated dramatically by their MFEs and the ejected plasmas in the MFEs have average temperatures close to the pre-eruption BP plasmas and electron densities typically near 109 cm-3. These new observational results indicate that CBPs are more complex in dynamical evolution and magnetic structure than previously thought.

  18. Globe at Night - Sky Brightness Monitoring Network

    NASA Astrophysics Data System (ADS)

    Cheung, Sze Leung; Pun, Jason Chun Shing; SO, Chu-wing; Shibata, Yukiko; Walker, Constance Elaine; Agata, Hidehiko

    2015-08-01

    The Global at Night - Sky Brightness Monitoring Network (GaN-MN) is an international project for long-term monitoring of night sky conditions around the world. The GaN-MN consists of fixed monitoring stations each equipped with a Sky Quality Meter - Lensed Ethernet (SQM-LE), which is a specialized light sensor for night sky brightness (NSB) measurement. NSB data are continuously collected at high sampling frequency throughout the night, and these data will be instantly made available to the general public to provide a real-time snapshot of the global light pollution condition. A single data collection methodology, including data sampling frequency, data selection criteria, device design and calibration, and schemes for data quality control, was adopted to ensure uniformity in the data collected. This is essential for a systematic and global study of the level of light pollution. The data collected will also provide the scientific backbone in our efforts to contribute to dark sky conservation through education to the general public and policy makers. The GaN-MN project is endorsed by the IAU IYL Executive Committee Working Group as a major Cosmic Light program in the International Year of Light.

  19. A high brightness field emission display

    SciTech Connect

    Palevsky, A.

    1996-12-31

    The military requirement for avionics display performance requires that displays be legible with 10,000 foot-candles (fc) bright light shining into the pilot`s eyes, or 10,000 fc shining directly on the display. The contrast ratio under these conditions must be at least 4.66:1. In addition, instant-on operation is sought for temperatures as low as {minus}54 C. Currently these specifications can barely be met by monochrome CRTs whose use is counter-indicated by other factors. No color display can achieve optimum performance in the areas mentioned, nor do any current contenders, primarily AMLCD`s, have any prospects of achieving full compliance. The FED being developed by Raytheon shows strong promise of being able to achieve the brightness and contrast ratios desired. The FED is also inherently able to provide instant-on functionality at any terrestrial temperature and does not require any heating at low temperatures. The technical objective of the Raytheon development program is to develop a high performance, full color, FED Panel that meets the performance and environmental operating condition requirements specified for military and other high performance display applications.

  20. Sublimation in bright spots on (1) Ceres

    NASA Astrophysics Data System (ADS)

    Nathues, A.; Hoffmann, M.; Schaefer, M.; Le Corre, L.; Reddy, V.; Platz, T.; Cloutis, E. A.; Christensen, U.; Kneissl, T.; Li, J.-Y.; Mengel, K.; Schmedemann, N.; Schaefer, T.; Russell, C. T.; Applin, D. M.; Buczkowski, D. L.; Izawa, M. R. M.; Keller, H. U.; O'Brien, D. P.; Pieters, C. M.; Raymond, C. A.; Ripken, J.; Schenk, P. M.; Schmidt, B. E.; Sierks, H.; Sykes, M. V.; Thangjam, G. S.; Vincent, J.-B.

    2015-12-01

    The dwarf planet (1) Ceres, the largest object in the main asteroid belt with a mean diameter of about 950 kilometres, is located at a mean distance from the Sun of about 2.8 astronomical units (one astronomical unit is the Earth-Sun distance). Thermal evolution models suggest that it is a differentiated body with potential geological activity. Unlike on the icy satellites of Jupiter and Saturn, where tidal forces are responsible for spewing briny water into space, no tidal forces are acting on Ceres. In the absence of such forces, most objects in the main asteroid belt are expected to be geologically inert. The recent discovery of water vapour absorption near Ceres and previous detection of bound water and OH near and on Ceres (refs 5, 6, 7) have raised interest in the possible presence of surface ice. Here we report the presence of localized bright areas on Ceres from an orbiting imager. These unusual areas are consistent with hydrated magnesium sulfates mixed with dark background material, although other compositions are possible. Of particular interest is a bright pit on the floor of crater Occator that exhibits probable sublimation of water ice, producing haze clouds inside the crater that appear and disappear with a diurnal rhythm. Slow-moving condensed-ice or dust particles may explain this haze. We conclude that Ceres must have accreted material from beyond the ‘snow line’, which is the distance from the Sun at which water molecules condense.

  1. Sublimation in bright spots on (1) Ceres.

    PubMed

    Nathues, A; Hoffmann, M; Schaefer, M; Le Corre, L; Reddy, V; Platz, T; Cloutis, E A; Christensen, U; Kneissl, T; Li, J-Y; Mengel, K; Schmedemann, N; Schaefer, T; Russell, C T; Applin, D M; Buczkowski, D L; Izawa, M R M; Keller, H U; O'Brien, D P; Pieters, C M; Raymond, C A; Ripken, J; Schenk, P M; Schmidt, B E; Sierks, H; Sykes, M V; Thangjam, G S; Vincent, J-B

    2015-12-10

    The dwarf planet (1) Ceres, the largest object in the main asteroid belt with a mean diameter of about 950 kilometres, is located at a mean distance from the Sun of about 2.8 astronomical units (one astronomical unit is the Earth-Sun distance). Thermal evolution models suggest that it is a differentiated body with potential geological activity. Unlike on the icy satellites of Jupiter and Saturn, where tidal forces are responsible for spewing briny water into space, no tidal forces are acting on Ceres. In the absence of such forces, most objects in the main asteroid belt are expected to be geologically inert. The recent discovery of water vapour absorption near Ceres and previous detection of bound water and OH near and on Ceres (refs 5-7) have raised interest in the possible presence of surface ice. Here we report the presence of localized bright areas on Ceres from an orbiting imager. These unusual areas are consistent with hydrated magnesium sulfates mixed with dark background material, although other compositions are possible. Of particular interest is a bright pit on the floor of crater Occator that exhibits probable sublimation of water ice, producing haze clouds inside the crater that appear and disappear with a diurnal rhythm. Slow-moving condensed-ice or dust particles may explain this haze. We conclude that Ceres must have accreted material from beyond the 'snow line', which is the distance from the Sun at which water molecules condense.

  2. Coronal bright points associated with minifilament eruptions

    SciTech Connect

    Hong, Junchao; Jiang, Yunchun; Yang, Jiayan; Bi, Yi; Li, Haidong; Yang, Bo; Yang, Dan

    2014-12-01

    Coronal bright points (CBPs) are small-scale, long-lived coronal brightenings that always correspond to photospheric network magnetic features of opposite polarity. In this paper, we subjectively adopt 30 CBPs in a coronal hole to study their eruptive behavior using data from the Atmospheric Imaging Assembly (AIA) and the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory. About one-quarter to one-third of the CBPs in the coronal hole go through one or more minifilament eruption(s) (MFE(s)) throughout their lifetimes. The MFEs occur in temporal association with the brightness maxima of CBPs and possibly result from the convergence and cancellation of underlying magnetic dipoles. Two examples of CBPs with MFEs are analyzed in detail, where minifilaments appear as dark features of a cool channel that divide the CBPs along the neutral lines of the dipoles beneath. The MFEs show the typical rising movements of filaments and mass ejections with brightenings at CBPs, similar to large-scale filament eruptions. Via differential emission measure analysis, it is found that CBPs are heated dramatically by their MFEs and the ejected plasmas in the MFEs have average temperatures close to the pre-eruption BP plasmas and electron densities typically near 10{sup 9} cm{sup –3}. These new observational results indicate that CBPs are more complex in dynamical evolution and magnetic structure than previously thought.

  3. Sublimation in bright spots on (1) Ceres.

    PubMed

    Nathues, A; Hoffmann, M; Schaefer, M; Le Corre, L; Reddy, V; Platz, T; Cloutis, E A; Christensen, U; Kneissl, T; Li, J-Y; Mengel, K; Schmedemann, N; Schaefer, T; Russell, C T; Applin, D M; Buczkowski, D L; Izawa, M R M; Keller, H U; O'Brien, D P; Pieters, C M; Raymond, C A; Ripken, J; Schenk, P M; Schmidt, B E; Sierks, H; Sykes, M V; Thangjam, G S; Vincent, J-B

    2015-12-10

    The dwarf planet (1) Ceres, the largest object in the main asteroid belt with a mean diameter of about 950 kilometres, is located at a mean distance from the Sun of about 2.8 astronomical units (one astronomical unit is the Earth-Sun distance). Thermal evolution models suggest that it is a differentiated body with potential geological activity. Unlike on the icy satellites of Jupiter and Saturn, where tidal forces are responsible for spewing briny water into space, no tidal forces are acting on Ceres. In the absence of such forces, most objects in the main asteroid belt are expected to be geologically inert. The recent discovery of water vapour absorption near Ceres and previous detection of bound water and OH near and on Ceres (refs 5-7) have raised interest in the possible presence of surface ice. Here we report the presence of localized bright areas on Ceres from an orbiting imager. These unusual areas are consistent with hydrated magnesium sulfates mixed with dark background material, although other compositions are possible. Of particular interest is a bright pit on the floor of crater Occator that exhibits probable sublimation of water ice, producing haze clouds inside the crater that appear and disappear with a diurnal rhythm. Slow-moving condensed-ice or dust particles may explain this haze. We conclude that Ceres must have accreted material from beyond the 'snow line', which is the distance from the Sun at which water molecules condense. PMID:26659183

  4. At Bright Band Inside Victoria Crater

    NASA Technical Reports Server (NTRS)

    2007-01-01

    A layer of light-toned rock exposed inside Victoria Crater in the Meridiani Planum region of Mars appears to mark where the surface was at the time, many millions of years ago, when an impact excavated the crater. NASA's Mars Exploration Rover Opportunity drove to this bright band as the science team's first destination for the rover during investigations inside the crater.

    Opportunity's left front hazard-identification camera took this image just after the rover finished a drive of 2.25 meters (7 feet, 5 inches) during the rover's 1,305th Martian day, or sol, (Sept. 25, 2007). The rocks beneath the rover and its extended robotic arm are part of the bright band.

    Victoria Crater has a scalloped shape of alternating alcoves and promontories around the crater's circumference. Opportunity descended into the crater two weeks earlier, within an alcove called 'Duck Bay.' Counterclockwise around the rim, just to the right of the arm in this image, is a promontory called 'Cabo Frio.'

  5. Wavelet dispersion and bright-spot detection

    SciTech Connect

    Luh, P.C.

    1989-03-01

    Since Ostrander in 1984 showed that the variations in reflectivity as a function of offset can be used for bright-spot validation, it has been known that these variations are often sensitive to errors in acquisition and processing of prestack seismic data. This presentation shows that even with perfect measurements, the bright-spot signal will be altered because the wavelet disperses as it propagates through an attenuating overburden. This in turn affects the slope term of the amplitude vs. offset (AVO) variation. For any single reflector, the dispersion of its propagated signal as a function of offset can be decomposed into a product of the vertical and horizontal dispersions. The vertical dispersion is the dispersion that a zero-offset arrival suffers through its propagation, and the horizontal dispersion is the additional dispersion that a nonzero-offset arrival must suffer further because of the residual normal moveout time. Numerical examples using a frequency-independent attenuation law show that even for a relatively high-Q or low-loss overburden, the horizontal dispersion alone can distort the AVO signal. This distortion cannot be taken care of by velocity analysis. A preferred method to overcome the dispersion effect would be to apply Q compensation on prestack seismic data before movement.

  6. Synchrotron brightness distribution of turbulent radio jets

    NASA Technical Reports Server (NTRS)

    Henriksen, R. N.; Bridle, A. H.; Chan, K. L.

    1981-01-01

    Radio jets are considered as turbulent mixing regions and it is proposed that the essential small scale viscous dissipation in these jets is by emission of MHD waves and by their subsequent strong damping due, at least partly, to gyro-resonant acceleration of supra-thermal particles. A formula relating the synchrotron surface brightness of a radio jet to the turbulent power input is deduced from physical postulates, and is tested against the data for NGC315 and 3C31 (NGC383). The predicted brightness depends essentially on the collimation behavior of the jet, and, to a lesser extent, on the CH picture of a 'high' nozzle with accelerating flow. The conditions for forming a large scale jet at a high nozzle from a much smaller scale jet are discussed. The effect of entrainment on the prediction is discussed with the use of similarity solutions. Although entrainment is inevitably associated with the turbulent jet, it may or may not be a dominant factor depending on the ambient density profile.

  7. Evaluation of HWRF Synthetic Satellite Brightness Temperatures

    NASA Astrophysics Data System (ADS)

    Musgrave, K. D.; Knaff, J. A.; Slocum, C. J.; Grasso, L. D.; Demaria, M.

    2014-12-01

    The regions within and surrounding tropical cyclones (TCs) tend to be devoid of in situ measurements, making satellite observations particularly useful within these data sparse regions. Forecasters and researchers have used these observations for decades to evaluate and understand the structural evolution of TCs. Synthetic satellite brightness temperatures are produced by the Hurricane Weather Research and Forecast (HWRF) model using the Community Radiative Transfer Model (CRTM) and provided for four different channels corresponding to channels available from the Geostationary Operational Environmental Satellite (GOES). This presentation will focus on the comparison between the HWRF synthetic satellite brightness temperatures and the observed GOES-13 10.7μm (infrared) and 6.48μm (water vapor) bands, highlighting the differences between the large-scale and storm-scale environments, and evaluating the structure of TCs as represented by the simulated satellite imagery. Disclaimer: The views, opinions, and findings contained in this article are those of the authors and should not be construed as an official National Oceanic and Atmospheric Administration (NOAA) or U.S. Government position, policy, or decision.

  8. Bright crater outflows: Possible emplacement mechanisms

    NASA Technical Reports Server (NTRS)

    Chadwick, D. John; Schaber, Gerald G.; Strom, Robert G.; Duval, Darla M.

    1992-01-01

    Lobate features with a strong backscatter are associated with 43 percent of the impact craters cataloged in Magellan's cycle 1. Their apparent thinness and great lengths are consistent with a low-viscosity material. The longest outflow yet identified is about 600 km in length and flows from the 90-km-diameter crater Addams. There is strong evidence that the outflows are largely composed of impact melt, although the mechanisms of their emplacement are not clearly understood. High temperatures and pressures of target rocks on Venus allow for more melt to be produced than on other terrestrial planets because lower shock pressures are required for melting. The percentage of impact craters with outflows increases with increasing crater diameter. The mean diameter of craters without outflows is 14.4 km, compared with 27.8 km for craters with outflows. No craters smaller than 3 km, 43 percent of craters in the 10- to 30-km-diameter range, and 90 percent in the 80- to 100-km-diameter range have associated bright outflows. More melt is produced in the more energetic impact events that produce larger craters. However, three of the four largest craters have no outflows. We present four possible mechanisms for the emplacement of bright outflows. We believe this 'shotgun' approach is justified because all four mechanisms may indeed have operated to some degree.

  9. Study of the brightness of trumpet tones

    NASA Astrophysics Data System (ADS)

    Poirson, Emilie; Petiot, Jean-François; Gilbert, Joël

    2005-10-01

    This study focuses on a particular attribute of trumpet tones, the brightness, and on the physical characteristics of the instrument thought to govern its magnitude. On the one hand, an objective study was carried out with input impedance measurements, and, on the other hand, a subjective study with hearing tests and a panel of subjects. To create a set of different trumpets a variable depth mouthpiece was developed whose depth can be easily and continuously adjusted from ``deep'' to ``shallow.'' Using this mouthpiece and the same trumpet, several instruments were generated which may be played in three ways: (i) by a musician, (ii) by an artificial mouth, and (iii) using physical modeling simulations. The influence of the depth of the mouthpiece on the perception of the trumpet's tones was investigated, and the ability of a musician, the artificial mouth, or physical modeling simulations to demonstrate perceptively noticeable differences was assessed. Physical characteristics extracted from the impedance curves are finally proposed to explain the brightness of trumpet tones. As a result, the physical modeling simulations now seem to be mature enough to exhibit coherent and subtle perceptual differences between tones. This opens the door to virtual acoustics for instrument makers.

  10. Generation and characterization of mouse monoclonal antibodies against NS4B protein of dengue virus.

    PubMed

    Xie, Xuping; Zou, Jing; Wang, Qing-Yin; Noble, Christian G; Lescar, Julien; Shi, Pei-Yong

    2014-02-01

    Dengue virus (DENV) non-structural protein 4B (NS4B) has been demonstrated to be an attractive antiviral target. Due to its nature as an integral membrane protein, NS4B remains poorly characterized. In this study, we generated and characterized two monoclonal antibodies (mAb) that selectively bind to DENV NS4B protein. One mAb, 10-3-7, is specific for DENV-2 NS4B, and its epitope was mapped to residues 5-15 of NS4B. The other mAb, 44-4-7, cross-reacts with all the four serotypes of DENV NS4B, and its epitope was mapped to residues 141-147 of NS4B. Using the mAbs, we probed the intracellular orientation of the epitopes of NS4B by an epitope accessibility assay. The results showed that the N-terminus of NS4B is located in the ER lumen, whereas amino acids 130-148 of NS4B are located in the cytosol. The study demonstrates that the two anti-NS4B mAbs will be useful for future structural and functional analyses of DENV NS4B. PMID:24503088

  11. A comparative biochemical analysis of the NS2B(H)-NS3pro protease complex from four dengue virus serotypes.

    PubMed

    Iempridee, Tawin; Thongphung, Ratchanu; Angsuthanasombat, Chanan; Katzenmeier, Gerd

    2008-01-01

    The two-component protease NS2B-NS3 of dengue virus mediates proteolytic processing of the polyprotein precursor and therefore represents a target for the development of antiviral drugs. The amino acid sequences of the NS3 serine protease and the NS2B cofactor exhibit relatively low degrees of conservation among the 4 serotypes thus suggesting that differences in enzyme activity exist which could modulate their susceptibility to future protease inhibitors. In this study we have addressed the question of functional similarity among the NS2B(H)-NS3pro proteases from 4 dengue virus serotypes by employing a uniform approach to clone, purify and assay proteolytic activity of these enzymes. Significant differences were observed for patterns of protein formation and expression levels in the E. coli host. Renaturation of the NS2B(H)-NS3pro precursors from dengue virus serotypes 2, 3 and 4 mediated by artificial chaperone-assisted refolding yielded enzymatically active proteases, whereas the enzyme from serotype 1 was obtained as soluble protein. Kinetic experiments using the GRR-amc substrate revealed comparable K(m) values while k(cat) values as obtained by active-site titration experiments displayed minor variations. Denaturation experiments demonstrated significant differences in half-life of the NS3 proteases from serotypes 2, 3 and 4 at 50 degrees C, whereas pH optima for all 4 enzymes were comparable.

  12. LARGER PLANET RADII INFERRED FROM STELLAR ''FLICKER'' BRIGHTNESS VARIATIONS OF BRIGHT PLANET-HOST STARS

    SciTech Connect

    Bastien, Fabienne A.; Stassun, Keivan G.; Pepper, Joshua

    2014-06-10

    Most extrasolar planets have been detected by their influence on their parent star, typically either gravitationally (the Doppler method) or by the small dip in brightness as the planet blocks a portion of the star (the transit method). Therefore, the accuracy with which we know the masses and radii of extrasolar planets depends directly on how well we know those of the stars, the latter usually determined from the measured stellar surface gravity, log g. Recent work has demonstrated that the short-timescale brightness variations ({sup f}licker{sup )} of stars can be used to measure log g to a high accuracy of ∼0.1-0.2 dex. Here, we use flicker measurements of 289 bright (Kepmag < 13) candidate planet-hosting stars with T {sub eff} = 4500-6650 K to re-assess the stellar parameters and determine the resulting impact on derived planet properties. This re-assessment reveals that for the brightest planet-host stars, Malmquist bias contaminates the stellar sample with evolved stars: nearly 50% of the bright planet-host stars are subgiants. As a result, the stellar radii, and hence the radii of the planets orbiting these stars, are on average 20%-30% larger than previous measurements had suggested.

  13. Does Stevens's Power Law for Brightness Extend to Perceptual Brightness Averaging?

    ERIC Educational Resources Information Center

    Bauer, Ben

    2009-01-01

    Stevens's power law ([Psi][infinity][Phi][beta]) captures the relationship between physical ([Phi]) and perceived ([Psi]) magnitude for many stimulus continua (e.g., luminance and brightness, weight and heaviness, area and size). The exponent ([beta]) indicates whether perceptual magnitude grows more slowly than physical magnitude ([beta] less…

  14. The 150 ns detector project: Prototype preamplifier results

    NASA Astrophysics Data System (ADS)

    Warburton, W. K.; Russell, S. R.; Kleinfelder, Stuart A.

    1994-08-01

    The long-term goal of the 150 ns detector project is to develop a pixel area detector capable of 6 MHz frame rates (150 ns/frame). Our milestones toward this goal are: a single pixel, 1×256 1D and 8×8 2D detectors, 256×256 2D detectors and, finally, 1024 × 1024 2D detectors. The design strategy is to supply a complete electronics chain (resetting preamp, selectable gain amplifier, analog-to-digital converter (ADC), and memory) for each pixel. In the final detectors these will all be custom integrated circuits. The front-end preamplifiers are integrated first, since their design and performance are the most unusual and also critical to the project's success. Similarly, our early work is concentrated on devising and perfecting detector structures. In this paper we demonstrate the performance of prototypes of our integrated preamplifiers. While the final design will have 64 preamps to a chip, including a switchable gain stage, the prototypes were integrated 8 channels to a "Tiny Chip" and tested in 4 configurations (feedback capacitor Cf equal 2.5 or 4.0 pF, output directly or through a source follower). These devices have been tested thoroughly for reset settling times, gain, linearity, and electronic noise. They generally work as designed, being fast enough to easily integrate detector charge, settle, and reset in 150 ns. Gain and linearity appear to be acceptable. Current values of electronic noise, in double-sampling mode, are about twice the design goal of {2}/{3} of a single photon at 6 keV. We expect this figure to improve with the addition of the onboard amplifier stage and improved packaging. Our next test chip will include these improvements and allow testing with our first detector samples, which will be 1×256 (50 μm wide pixels) and 8×8 (1 mm 2 pixels) element detector on 1 mm thick silicon.

  15. VM version of INTERLAN's NS4240 Xerox ITP Network Software

    SciTech Connect

    Frese, H.; Cottrell, R.L.; Downey, T.

    1986-04-01

    This manual describes Stanford Linear Accelerator Center's VM adaptation of INTERLAN's NS4240 ITP Network Software. The ITP Network Software is an implementation of the Xerox Network Systems Internet Transport Protocols. The ITP Network Software runs under the VM/SP operating system. This manual assumes familiarity with the use of the VM operating system. The user is also expected to have experience in assembling, linking, and running application programs on a system. The user should be familiar with the concepts of computer networking and have an understanding of the more specific concepts of Ethernet-based networks. 10 refs.

  16. Cyclosporine Inhibits a Direct Interaction between Cyclophilins and Hepatitis C NS5A

    PubMed Central

    Striker, Rob

    2010-01-01

    Background Hepatitis C Virus (HCV) infection is a leading indication for liver transplantation. HCV infection reoccurs almost universally post transplant, decreasing both graft longevity and patient survival. The immunosuppressant, cyclosporine A (CsA) has potent anti-HCV activity towards both HCV replicons and the genotype 2a cell culture infectious virus. Previously, we isolated mutations in the 1bN replicon with less sensitivity to CsA that mapped to both NS5A and NS5B regions of the virus. Mutations in NS5A alone conferred decreased CsA susceptibility regardless of NS5B mutations. Methodology/Principal Findings We examined the mechanisms by which NS5A mutations contribute to CsA resistance and if they are strain dependent. Using in vitro mutagenesis, the amino acid position 321 mutation of NS5A was restored to the wild-type tyrosine residue conferring partial CsA susceptibility on the mutant replicon. The 321 mutation also alters CsA susceptibility of the JFH cell culture virus. Additionally, we demonstrated a novel CsA-sensitive interaction between NS5A and both cyclophilin A and B. Both the mutant NS5A and wild type NS5A bind cyclophilin in vitro. The NS5A: cyclophilin interaction requires both the NS5A region identified by the resistance mutants and cyclophilin catalytic residues. In cell culture, NS5A from CsA resistant mutant has an enhanced interaction with cyclophilin B. Additionally; NS5B facilitates a stronger binding of mutant NS5A to endogenous cyclophilin B than wild-type in cell culture. Conclusions/Significance Collectively, this data suggests direct interactions between cyclophilins and NS5A are critical to understand for optimal use of cyclophilin inhibitors in anti-HCV therapy. PMID:20352119

  17. New Distant Comet Headed for Bright Encounter

    NASA Astrophysics Data System (ADS)

    1995-08-01

    How Impressive Will Comet Hale-Bopp Become in 1997 ? A very unusual comet was discovered last month, on its way from the outer reaches of the solar system towards the Sun. Although it is still situated beyond the orbit of Jupiter, it is so bright that it can be observed in even small telescopes. It has been named `Hale-Bopp' after the discoverers and is already of great interest to cometary astronomers. No less than seven telescopes have been used at the ESO La Silla observatory for the first observations of the new object. Together with data gathered at other sites, their aim is to elucidate the nature of this comet and also to determine whether there is reason to hope that it will become a bright and beautiful object in the sky from late 1996 and well into 1997. Further observations are now being planned at ESO and elsewhere to monitor closely the behaviour of this celestial visitor during the coming months. Discovery circumstances The comet was discovered on 23 July 1995, nearly simultaneously by two American amateur astronomers, Alan Hale of Cloudcroft (New Mexico) and Thomas Bopp of Glendale (Arizona). Although the chronology is slightly uncertain, it appears that Hale first saw it some 10 - 20 minutes before Bopp, at 06:10 - 06:15 UT on that day. In any case, he informed the IAU Central Bureau for Astronomical Telegrams (CBAT) in Cambridge (Massachussetts) about his discovery by email already at 06:50 UT, while Bopp's message was filed more than 2 hours later, after he had driven back to his home, 140 km from where he had been observing. Upon receipt of these messages, Brian Marsden at the CBAT assigned the designation `1995 O1' (indicating that it is the first comet found in the second half of July 1995). After further sightings had been made by other observers, and according to the venerable astronomical tradition, the new object was named after the discoverers. The magnitude, reported as 10.5 by Hale, is not unusual for a comet that is discovered within

  18. In vitro Splicing of Influenza Viral NS1 mRNA and NS1-β -globin Chimeras: Possible Mechanisms for the Control of Viral mRNA Splicing

    NASA Astrophysics Data System (ADS)

    Plotch, Stephen J.; Krug, Robert M.

    1986-08-01

    In influenza virus-infected cells, the splicing of the viral NS1 mRNA catalyzed by host nuclear enzymes is controlled so that the steady-state amount of the spliced NS2 mRNA is only 5-10% of that of the unspliced NS1 mRNA. Here we examine the splicing of NS1 mRNA in vitro, using nuclear extracts from HeLa cells. We show that in addition to its consensus 5' and 3' splice sites, NS1 mRNA has an intron branch-point adenosine residue that was functional in lariat formation. Nonetheless, this RNA was not detectably spliced in vitro under conditions in which a human β -globin precursor was efficiently spliced. Using chimeric RNA precursors containing both NS1 and β -globin sequences, we show that the NS1 5' splice site was effectively utilized by the β -globin branch-point sequence and 3' splice site to form a spliced RNA, whereas the NS1 3' splice site did not function in detectable splicing in vitro, even in the presence of the β -globin branch-point sequence or in the presence of both the branch-point sequence and 5' exon and splice site from β -globin With the chimeric precursors that were not detectably spliced, as with NS1 mRNA itself, a low level of a lariat structure containing only intron and not 3' exon sequences was formed. The inability of the consensus 3' splice site of NS1 mRNA to function effectively in in vitro splicing suggests that this site is structurally inaccessible to components of the splicing machinery. Based on these results, we propose two mechanisms whereby NS1 mRNA splicing in infected cells is controlled via the accessibility of its 3' splice site.

  19. Antiviral potential of 4-hydroxypanduratin A, secondary metabolite of Fingerroot, Boesenbergia pandurata (Schult.), towards Japanese Encephalitis virus NS2B/NS3 protease.

    PubMed

    Seniya, Chandrabhan; Mishra, Harshal; Yadav, Ajay; Sagar, Nitin; Chaturvedi, Babita; Uchadia, Kuldeep; Wadhwa, Gulshan

    2013-01-01

    4-hydroxypanduratin A is a secondary metabolite of Boesenbergia pandurata Schult. (Fingerroot) plant with various pharmacological activities such as neuroprotective, potent antioxidant, antibacterial and antifungal. Flaviviral NS2B/NS3 protease activity is essential for polyprotein processing and viral replication for Japanese Encephalitis Virus (JEV), a major cause of Acute Encephaltis in Asia. Inhibition of formation of this complex by arresting the binding of NS2B with NS3 would reduce the enzyme's activity to meager proportions and hence would prevent further viral proliferation. The automated 3D structure of NS2B protein of the JEV GP78 was predicted based on the sequence-to-structure-to-function paradigm using I-TASSER and the function of NS2B protein was inferred by matching to other known proteins. The stereochemical quality of predicted structure was checked by PROCHECK. The antiviral activity of 4-hydroxypanduratin A against NS2B protein as a potential drug has been elucidated in this paper. Docking simulation analysis showed 4-hydroxypanduratin A as potential inhibitor of NS2B protein/cofactor which is necessary for NS3 protease activity. 220 derivatives of 4-hydroxypanduratin A were virtually screened with rigid criteria of Lipinski's rule of 5 using Autodock4.2. 4-hydroxypanduratin A was found interacting with target hydrophilic domain in NS2B protein by two Hbonds (Gly80 and Asp81) with active residues, several hydrophobic interactions, Log P value of 5.6, inhibition constant (Ki) of 51.07nM and lowest binding energy of -9.95Kcal/Mol. Hence, 4-hydroxypanduratin A targeted to Site 2 will have sufficient profound effect to inhibit protease activity to abrogate viral replication. It could be a promising potential drug candidate for JEV infections using NS2B Site 2 as a Drug target.

  20. Influenza C virus NS1 protein upregulates the splicing of viral mRNAs.

    PubMed

    Muraki, Yasushi; Furukawa, Takatoshi; Kohno, Yoshihiko; Matsuzaki, Yoko; Takashita, Emi; Sugawara, Kanetsu; Hongo, Seiji

    2010-02-01

    Pre-mRNAs of the influenza A virus M and NS genes are poorly spliced in virus-infected cells. By contrast, in influenza C virus-infected cells, the predominant transcript from the M gene is spliced mRNA. The present study was performed to investigate the mechanism by which influenza C virus M gene-specific mRNA (M mRNA) is readily spliced. The ratio of M1 encoded by a spliced M mRNA to CM2 encoded by an unspliced M mRNA in influenza C virus-infected cells was about 10 times larger than that in M gene-transfected cells, suggesting that a viral protein(s) other than M gene translational products facilitates viral mRNA splicing. RNase protection assays showed that the splicing of M mRNA in infected cells was much higher than that in M gene-transfected cells. The unspliced and spliced mRNAs of the influenza C virus NS gene encode two nonstructural (NS) proteins, NS1(C/NS1) and NS2(C/NS2), respectively. The introduction of premature translational termination into the NS gene, which blocked the synthesis of the C/NS1 and C/NS2 proteins, drastically reduced the splicing of NS mRNA, raising the possibility that C/NS1 or C/NS2 enhances viral mRNA splicing. The splicing of influenza C virus M mRNA was increased by coexpression of C/NS1, whereas it was reduced by coexpression of the influenza A virus NS1 protein (A/NS1). The splicing of influenza A virus M mRNA was also increased by coexpression of C/NS1, though it was inhibited by that of A/NS1. These results suggest that influenza C virus NS1, but not A/NS1, can upregulate viral mRNA splicing.

  1. Folding Proteins at 500 ns/hour with Work Queue

    PubMed Central

    Abdul-Wahid, Badi’; Yu, Li; Rajan, Dinesh; Feng, Haoyun; Darve, Eric; Thain, Douglas; Izaguirre, Jesús A.

    2014-01-01

    Molecular modeling is a field that traditionally has large computational costs. Until recently, most simulation techniques relied on long trajectories, which inherently have poor scalability. A new class of methods is proposed that requires only a large number of short calculations, and for which minimal communication between computer nodes is required. We considered one of the more accurate variants called Accelerated Weighted Ensemble Dynamics (AWE) and for which distributed computing can be made efficient. We implemented AWE using the Work Queue framework for task management and applied it to an all atom protein model (Fip35 WW domain). We can run with excellent scalability by simultaneously utilizing heterogeneous resources from multiple computing platforms such as clouds (Amazon EC2, Microsoft Azure), dedicated clusters, grids, on multiple architectures (CPU/GPU, 32/64bit), and in a dynamic environment in which processes are regularly added or removed from the pool. This has allowed us to achieve an aggregate sampling rate of over 500 ns/hour. As a comparison, a single process typically achieves 0.1 ns/hour. PMID:25540799

  2. X-ray structures of NS1 effector domain mutants.

    PubMed

    Xia, Shuangluo; Robertus, Jon D

    2010-02-15

    The influenza A virus nonstructural protein NS1 is a multifunctional dimeric protein that acts as a potent inhibitor of the host cellular antiviral state. The C-terminal effector domain of NS1 binds host proteins, including CPSF30, and is a target for the development of new antiviral drugs. Here we present crystallographic structures of two mutant effector domains, W187Y and W187A, of influenza A/Udorn/72 virus. Unlike wild-type, the mutants behave exclusively as monomers in solution based on gel filtration data and light scattering. The W187Y mutant is able to bind CPSF30 with a binding affinity close to the wild-type protein; that is, it retains a receptor site for aromatic ligands nearly identical to the wild-type. Therefore, this monomeric mutant protein could serve as a drug target for a high throughput inhibitor screening assays, since its binding pocket is unoccupied in solution and potentially more accessible to small molecule ligands.

  3. Dengue virus RNA polymerase NS5: a potential therapeutic target?

    PubMed

    Rawlinson, Stephen M; Pryor, Melinda J; Wright, Peter J; Jans, David A

    2006-12-01

    Dengue fever (DF)/dengue haemorrhagic fever (DHF) is the most common arthropod-borne viral infection, where it is now estimated that 2.5-3 billion people world-wide are at risk of infection. Currently there is no available treatment, in the form of vaccine or drug, making eradication of the mosquito vector the only viable control measure, which has proved costly and of limited success. There are a number of different vaccines undergoing testing, but whilst a dengue vaccine is clearly desirable, there are several issues which make live-attenuated vaccines problematic. These include the phenomenon of antibody-dependent enhancement (ADE) and the possibility of recombination of attenuated vaccine strains with wild-type flavivirus members reverting vaccines to a virulent form. Until we gain a better understanding of these issues and their associated risks, the safety of any live dengue vaccine cannot be assured. It therefore may be safer and more feasible for therapeutic-based approaches to be developed as an alternative to live vaccines. As our understanding of dengue molecular biology expands, new potential targets for drugs are emerging. One of the most promising is the dengue non-structural protein 5 (NS5), the largest and most highly conserved of the dengue proteins. This review examines the unique properties of NS5, including its functions, interactions, subcellular localisation and regulation, and looks at ways in which some of these may be exploited in our quest for effective drugs.

  4. WIDGET: Ultra Bright Optical Transients Search

    NASA Astrophysics Data System (ADS)

    Urata, Y.; Lin, H.-M.; Tashiro, M.; Widget Team

    2010-12-01

    The WIDeField telescope for Gamma-ray burst Early Timing (WIDGET) is used for a fully automated, ultra-wide-field survey aimed at detecting the prompt optical emission associated with Gamma-ray Bursts (GRB). WIDGET surveys the HETE-2 and Swift/BAT pointing directions covering a total field of view of 62x62 degree every 10 seconds using an unfiltered system. This monitoring survey allows exploration of the optical emission before the gamma-ray trigger. At the same time, WIDGET is the quite unique instrument to explore ultra-bright optical transients with high time resolution. Combination with the MAXI survey, WIDGET has higher potential to make simultaneous discoveries of optical counterparts for high energy transients. We developed the serendipitous transient finding pipeline. We report the preliminary results.

  5. Brightness temperature for 166 radio sources

    NASA Astrophysics Data System (ADS)

    Fan, Jun-Hui; Huang, Yong; Yuan, Yu-Hai; Yang, Jiang-He; Liu, Yi; Tao, Jun; Gao, Ying; Hua, Tong-Xu; Lin, Rui-Guang; Zhang, Jiang-Shui; Zhang, Jing-Yi; Qin, Yi-Ping

    2009-07-01

    Using the database of the University of Michigan Radio Astronomy Observatory (UMRAO) at three radio frequencies (4.8, 8 and 14.5 GHz), we determined the short-term variability timescales for 166 radio sources. The timescales are 0.15 d (2007+777) to 176.17 d (0528-250) with an average timescale of Δtobs = 17.1 ± 16.5 d for the whole sample. The timescales are used to calculate the brightness temperatures, TB. The value of log TB is in the range of log TB = 10.47 to 19.06 K. In addition, we also estimated the boosting factor for the sources. The correlation between the polarization and the Doppler factor is also discussed.

  6. High brightness angled cavity quantum cascade lasers

    SciTech Connect

    Heydari, D.; Bai, Y.; Bandyopadhyay, N.; Slivken, S.; Razeghi, M.

    2015-03-02

    A quantum cascade laser (QCL) with an output power of 203 W is demonstrated in pulsed mode at 283 K with an angled cavity. The device has a ridge width of 300 μm, a cavity length of 5.8 mm, and a tilt angle of 12°. The back facet is high reflection coated, and the front facet is anti-reflection coated. The emitting wavelength is around 4.8 μm. In distinct contrast to a straight cavity broad area QCL, the lateral far field is single lobed with a divergence angle of only 3°. An ultrahigh brightness value of 156 MW cm{sup −2 }sr{sup −1} is obtained, which marks the brightest QCL to date.

  7. Bright photoluminescent hybrid mesostructured silica nanoparticles.

    PubMed

    Miletto, Ivana; Bottinelli, Emanuela; Caputo, Giuseppe; Coluccia, Salvatore; Gianotti, Enrica

    2012-07-28

    Bright photoluminescent mesostructured silica nanoparticles were synthesized by the incorporation of fluorescent cyanine dyes into the channels of MCM-41 mesoporous silica. Cyanine molecules were introduced into MCM-41 nanoparticles by physical adsorption and covalent grafting. Several photoluminescent nanoparticles with different organic loadings have been synthesized and characterized by X-ray powder diffraction, high resolution transmission electron microscopy and nitrogen physisorption porosimetry. A detailed photoluminescence study with the analysis of fluorescence lifetimes was carried out to elucidate the cyanine molecules distribution within the pores of MCM-41 nanoparticles and the influence of the encapsulation on the photoemission properties of the guests. The results show that highly stable photoluminescent hybrid materials with interesting potential applications as photoluminescent probes for diagnostics and imaging can be prepared by both methods. PMID:22706523

  8. High brightness angled cavity quantum cascade lasers

    NASA Astrophysics Data System (ADS)

    Heydari, D.; Bai, Y.; Bandyopadhyay, N.; Slivken, S.; Razeghi, M.

    2015-03-01

    A quantum cascade laser (QCL) with an output power of 203 W is demonstrated in pulsed mode at 283 K with an angled cavity. The device has a ridge width of 300 μm, a cavity length of 5.8 mm, and a tilt angle of 12°. The back facet is high reflection coated, and the front facet is anti-reflection coated. The emitting wavelength is around 4.8 μm. In distinct contrast to a straight cavity broad area QCL, the lateral far field is single lobed with a divergence angle of only 3°. An ultrahigh brightness value of 156 MW cm-2 sr-1 is obtained, which marks the brightest QCL to date.

  9. Modelling Solar and Stellar Brightness Variabilities

    NASA Astrophysics Data System (ADS)

    Yeo, K. L.; Shapiro, A. I.; Krivova, N. A.; Solanki, S. K.

    2016-04-01

    Total and spectral solar irradiance, TSI and SSI, have been measured from space since 1978. This is accompanied by the development of models aimed at replicating the observed variability by relating it to solar surface magnetism. Despite significant progress, there remains persisting controversy over the secular change and the wavelength-dependence of the variation with impact on our understanding of the Sun's influence on the Earth's climate. We highlight the recent progress in TSI and SSI modelling with SATIRE. Brightness variations have also been observed for Sun-like stars. Their analysis can profit from knowledge of the solar case and provide additional constraints for solar modelling. We discuss the recent effort to extend SATIRE to Sun-like stars.

  10. Hybrid quantum repeater using bright coherent light.

    PubMed

    van Loock, P; Ladd, T D; Sanaka, K; Yamaguchi, F; Nemoto, Kae; Munro, W J; Yamamoto, Y

    2006-06-23

    We describe a quantum repeater protocol for long-distance quantum communication. In this scheme, entanglement is created between qubits at intermediate stations of the channel by using a weak dispersive light-matter interaction and distributing the outgoing bright coherent-light pulses among the stations. Noisy entangled pairs of electronic spin are then prepared with high success probability via homodyne detection and postselection. The local gates for entanglement purification and swapping are deterministic and measurement-free, based upon the same coherent-light resources and weak interactions as for the initial entanglement distribution. Finally, the entanglement is stored in a nuclear-spin-based quantum memory. With our system, qubit-communication rates approaching 100 Hz over 1280 km with fidelities near 99% are possible for reasonable local gate errors.

  11. Dark Skies, Bright Kids Year 7

    NASA Astrophysics Data System (ADS)

    Bittle, Lauren E.; Johnson, Kelsey E.; Borish, H. Jacob; Burkhardt, Andrew; Firebaugh, Ariel; Hancock, Danielle; Rochford Hayes, Christian; Linden, Sean; Liss, Sandra; Matthews, Allison; Prager, Brian; Pryal, Matthew; Sokal, Kimberly R.; Troup, Nicholas William; Wenger, Trey

    2016-01-01

    We present updates from our seventh year of operation including new club content, continued assessments, and our fifth annual Star Party. Dark Skies, Bright Kids (DSBK) is an entirely volunteer-run outreach organization based out of the Department of Astronomy at the University of Virginia. Our core mission is to enhance elementary science education and literacy in Central Virginia through fun, hands-on activities that introduce basic Astronomy concepts. Our primary focus is hosting an 8-10 week after-school astronomy club at underserved elementary and middle schools. Each week, DSBK volunteers take the role of coaches to introduce astronomy-related concepts ranging from the Solar System to galaxies to astrobiology, and to lead students in interactive learning activities. Another hallmark of DSBK is hosting our Annual Central Virginia Star Party, a free event open to the community featuring star-gazing and planetarium shows.

  12. The 2NS Translocation from Aegilops ventricosa Confers Resistance to the Triticum Pathotype of Magnaporthe oryzae

    PubMed Central

    Cruz, C.D.; Peterson, G.L.; Bockus, W.W.; Kankanala, P.; Dubcovsky, J.; Jordan, K.W.; Akhunov, E.; Chumley, F.; Baldelomar, F.D.; Valent, B.

    2016-01-01

    Wheat blast is a serious disease caused by the fungus Magnaporthe oryzae (Triticum pathotype) (MoT). The objective of this study was to determine the effect of the 2NS translocation from Aegilops ventricosa (Zhuk.) Chennav on wheat head and leaf blast resistance. Disease phenotyping experiments were conducted in growth chamber, greenhouse, and field environments. Among 418 cultivars of wheat (Triticum aestivum L.), those with 2NS had 50.4 to 72.3% less head blast than those without 2NS when inoculated with an older MoT isolate under growth chamber conditions. When inoculated with recently collected isolates, cultivars with 2NS had 64.0 to 80.5% less head blast. Under greenhouse conditions when lines were inoculated with an older MoT isolate, those with 2NS had a significant head blast reduction. With newer isolates, not all lines with 2NS showed a significant reduction in head blast, suggesting that the genetic background and/or environment may influence the expression of any resistance conferred by 2NS. However, when near-isogenic lines (NILs) with and without 2NS were planted in the field, there was strong evidence that 2NS conferred resistance to head blast. Results from foliar inoculations suggest that the resistance to head infection that is imparted by the 2NS translocation does not confer resistance to foliar disease. In conclusion, the 2NS translocation was associated with significant reductions in head blast in both spring and winter wheat. PMID:27814405

  13. Bright Soil Near 'McCool'

    NASA Technical Reports Server (NTRS)

    2006-01-01

    While driving eastward toward the northwestern flank of 'McCool Hill,' the wheels of NASA's Mars Exploration Rover Spirit churned up the largest amount of bright soil discovered so far in the mission. This image from Spirit's panoramic camera (Pancam), taken on the rover's 788th Martian day, or sol, of exploration (March 22, 2006), shows the strikingly bright tone and large extent of the materials uncovered.

    Several days earlier, Spirit's wheels unearthed a small patch of light-toned material informally named 'Tyrone.' In images from Spirit's panoramic camera, 'Tyrone' strongly resembled both 'Arad' and 'Paso Robles,' two patches of light-toned soils discovered earlier in the mission. Spirit found 'Paso Robles' in 2005 while climbing 'Cumberland Ridge' on the western slope of 'Husband Hill.' In early January 2006, the rover discovered 'Arad' on the basin floor just south of 'Husband Hill.' Spirit's instruments confirmed that those soils had a salty chemistry dominated by iron-bearing sulfates. Spirit's Pancam and miniature thermal emission spectrometer examined this most recent discovery, and researchers will compare its properties with the properties of those other deposits.

    These discoveries indicate that salty, light-toned soil deposits might be widely distributed on the flanks and valley floors of the 'Columbia Hills' region in Gusev Crater on Mars. The salts, which are easily mobilized and concentrated in liquid solution, may record the past presence of water. So far, these enigmatic materials have generated more questions than answers, however, and as Spirit continues to drive across this region in search of a safe winter haven, the team continues to formulate and test hypotheses to explain the rover's most fascinating recent discovery.

    This view is an approximately true-color rendering that combines separate images taken through the Pancam's 753-nanometer, 535-nanometer, and 432-nanometer filters.

  14. Statistical Properties of Solar Coronal Bright Points

    NASA Astrophysics Data System (ADS)

    Alipour, N.; Safari, H.

    2015-07-01

    Here, we aim to study the statistical properties (i.e., spatial, temporal, and magnetic structures) of extreme ultraviolet coronal bright points (CBPs) observed by SDO during a 4.4 yr period (2010 June 1 to 2014 October 31). We developed the automatic detection method for CBPs based on the machine-learning technique and Zernike image moments. The average number and the mean density of CBPs are estimated to be about 572 (per full disk image taken at 193 Å) and 1.9× {10}-4 Mm‑2, respectively. There is a negative correlation (‑0.7) between the number of CBPs and the number of sunspots. The size and lifetime frequency distribution of CBPs show the lognormal and power-law (exponent equal to ‑1.6) behaviors, respectively. The relationship between the lifetime and size of CBPs is clearly treated by a power-law function with an exponent equal to 0.13. Around 1.3% of the solar surface is covered by the bright cores of CBPs and 2.6% of that is covered by their total area. About 52% of CBPs have lifetimes of less than 20 minutes and the remaining 48% have mean lifetimes of 6 hr. More than 95% of CBPs with lifetimes of less than 20 hr and nine CBPs with lifetimes of more than 72 hr are detected. The average number of the new CBPs emerging every 45 s in the whole of the Sun is about 27 ± 3. The temporal self-affinity of the time series of CBPs that emerged, indexed by the Hurst exponent determined using both detrended fluctuation analysis and R/S analysis, is 0.78. This long-temporal correlation suggests that CBPs form a system of self-organized criticality.

  15. The GPM Common Calibrated Brightness Temperature Product

    NASA Technical Reports Server (NTRS)

    Stout, John; Berg, Wesley; Huffman, George; Kummerow, Chris; Stocker, Erich

    2005-01-01

    The Global Precipitation Measurement (GPM) project will provide a core satellite carrying the GPM Microwave Imager (GMI) and will use microwave observations from a constellation of other satellites. Each partner with a satellite in the constellation will have a calibration that meets their own requirements and will decide on the format to archive their brightness temperature (Tb) record in GPM. However, GPM multi-sensor precipitation algorithms need to input intercalibrated Tb's in order to avoid differences among sensors introducing artifacts into the longer term climate record of precipitation. The GPM Common Calibrated Brightness Temperature Product is intended to address this problem by providing intercalibrated Tb data, called "Tc" data, where the "c" stands for common. The precipitation algorithms require a Tc file format that is both generic and flexible enough to accommodate the different passive microwave instruments. The format will provide detailed information on the processing history in order to allow future researchers to have a record of what was done. The format will be simple, including the main items of scan time, latitude, longitude, and Tc. It will also provide spacecraft orientation, spacecraft location, orbit, and instrument scan type (cross-track or conical). Another simplification is to store data in real numbers, avoiding the ambiguity of scaled data. Finally, units and descriptions will be provided in the product. The format is built on the concept of a swath, which is a series of scans that have common geolocation and common scan geometry. Scan geometry includes pixels per scan, sensor orientation, scan type, and incidence angles. The Tc algorithm and data format are being tested using the pre-GPM Precipitation Processing System (PPS) software to generate formats and 1/0 routines. In the test, data from SSM/I, TMI, AMSR-E, and WindSat are being processed and written as Tc products.

  16. Enhancement of Virus Replication in An Influenza A Virus NS1-Expresssing 293 Cell Line.

    PubMed

    Zhu, Wu Yang; Tao, Xiao Yan; Lyu, Xin Jun; Yu, Peng Cheng; Lu, Zhuo Zhuang

    2016-03-01

    The nonstructural protein 1 (NS1) of influenza A virus, which is absent from the viral particle, but highly expressed in infected cells, strongly antagonizes the interferon (IFN)-mediated antiviral response. We engineered an NS1-expressing 293 (293-NS1) cell line with no response to IFN stimulation. Compared with the parental 293 cells, the IFN-nonresponsive 293-NS1 cells improved the growth capacity of various viruses, but the introduction of NS1 barely enhanced the propagation of Tahyna virus, a negative-strand RNA virus. In particular, fastidious enteric adenovirus that replicates poorly in 293 cells may grow more efficiently in 293-NS1 cells; thus, IFN-nonresponsive 293-NS1 cells might be of great value in diagnostic laboratories for the cultivation and isolation of human enteric adenoviruses.

  17. Enhancement of Virus Replication in An Influenza A Virus NS1-Expresssing 293 Cell Line.

    PubMed

    Zhu, Wu Yang; Tao, Xiao Yan; Lyu, Xin Jun; Yu, Peng Cheng; Lu, Zhuo Zhuang

    2016-03-01

    The nonstructural protein 1 (NS1) of influenza A virus, which is absent from the viral particle, but highly expressed in infected cells, strongly antagonizes the interferon (IFN)-mediated antiviral response. We engineered an NS1-expressing 293 (293-NS1) cell line with no response to IFN stimulation. Compared with the parental 293 cells, the IFN-nonresponsive 293-NS1 cells improved the growth capacity of various viruses, but the introduction of NS1 barely enhanced the propagation of Tahyna virus, a negative-strand RNA virus. In particular, fastidious enteric adenovirus that replicates poorly in 293 cells may grow more efficiently in 293-NS1 cells; thus, IFN-nonresponsive 293-NS1 cells might be of great value in diagnostic laboratories for the cultivation and isolation of human enteric adenoviruses. PMID:27109134

  18. New Distant Comet Headed for Bright Encounter

    NASA Astrophysics Data System (ADS)

    1995-08-01

    How Impressive Will Comet Hale-Bopp Become in 1997 ? A very unusual comet was discovered last month, on its way from the outer reaches of the solar system towards the Sun. Although it is still situated beyond the orbit of Jupiter, it is so bright that it can be observed in even small telescopes. It has been named `Hale-Bopp' after the discoverers and is already of great interest to cometary astronomers. No less than seven telescopes have been used at the ESO La Silla observatory for the first observations of the new object. Together with data gathered at other sites, their aim is to elucidate the nature of this comet and also to determine whether there is reason to hope that it will become a bright and beautiful object in the sky from late 1996 and well into 1997. Further observations are now being planned at ESO and elsewhere to monitor closely the behaviour of this celestial visitor during the coming months. Discovery circumstances The comet was discovered on 23 July 1995, nearly simultaneously by two American amateur astronomers, Alan Hale of Cloudcroft (New Mexico) and Thomas Bopp of Glendale (Arizona). Although the chronology is slightly uncertain, it appears that Hale first saw it some 10 - 20 minutes before Bopp, at 06:10 - 06:15 UT on that day. In any case, he informed the IAU Central Bureau for Astronomical Telegrams (CBAT) in Cambridge (Massachussetts) about his discovery by email already at 06:50 UT, while Bopp's message was filed more than 2 hours later, after he had driven back to his home, 140 km from where he had been observing. Upon receipt of these messages, Brian Marsden at the CBAT assigned the designation `1995 O1' (indicating that it is the first comet found in the second half of July 1995). After further sightings had been made by other observers, and according to the venerable astronomical tradition, the new object was named after the discoverers. The magnitude, reported as 10.5 by Hale, is not unusual for a comet that is discovered within

  19. A conserved predicted pseudoknot in the NS2A-encoding sequence of West Nile and Japanese encephalitis flaviviruses suggests NS1' may derive from ribosomal frameshifting

    PubMed Central

    Firth, Andrew E; Atkins, John F

    2009-01-01

    Japanese encephalitis, West Nile, Usutu and Murray Valley encephalitis viruses form a tight subgroup within the larger Flavivirus genus. These viruses utilize a single-polyprotein expression strategy, resulting in ~10 mature proteins. Plotting the conservation at synonymous sites along the polyprotein coding sequence reveals strong conservation peaks at the very 5' end of the coding sequence, and also at the 5' end of the sequence encoding the NS2A protein. Such peaks are generally indicative of functionally important non-coding sequence elements. The second peak corresponds to a predicted stable pseudoknot structure whose biological importance is supported by compensatory mutations that preserve the structure. The pseudoknot is preceded by a conserved slippery heptanucleotide (Y CCU UUU), thus forming a classical stimulatory motif for -1 ribosomal frameshifting. We hypothesize, therefore, that the functional importance of the pseudoknot is to stimulate a portion of ribosomes to shift -1 nt into a short (45 codon), conserved, overlapping open reading frame, termed foo. Since cleavage at the NS1-NS2A boundary is known to require synthesis of NS2A in cis, the resulting transframe fusion protein is predicted to be NS1-NS2AN-term-FOO. We hypothesize that this may explain the origin of the previously identified NS1 'extension' protein in JEV-group flaviviruses, known as NS1'. PMID:19196463

  20. Association of hepatitis C virus replication complexes with microtubules and actin filaments is dependent on the interaction of NS3 and NS5A.

    PubMed

    Lai, Chao-Kuen; Jeng, King-Song; Machida, Keigo; Lai, Michael M C

    2008-09-01

    The hepatitis C virus (HCV) RNA replication complex (RC), which is composed of viral nonstructural (NS) proteins and host cellular proteins, replicates the viral RNA genome in association with intracellular membranes. Two viral NS proteins, NS3 and NS5A, are essential elements of the RC. Here, by using immunoprecipitation and fluorescence resonance energy transfer assays, we demonstrated that NS3 and NS5A interact with tubulin and actin. Furthermore, immunofluorescence microscopy and electron microscopy revealed that HCV RCs were aligned along microtubules and actin filaments in both HCV replicon cells and HCV-infected cells. In addition, the movement of RCs was inhibited when microtubules or actin filaments were depolymerized by colchicine and cytochalasin B, respectively. Based on our observations, we propose that microtubules and actin filaments provide the tracks for the movement of HCV RCs to other regions in the cell, and the molecular interactions between RCs and microtubules, or RCs and actin filaments, are mediated by NS3 and NS5A. PMID:18562541

  1. Measurement of night sky brightness in southern Australia

    NASA Astrophysics Data System (ADS)

    Hampf, Daniel; Rowell, Gavin; Wild, Neville; Sudholz, Tristan; Horns, Dieter; Tluczykont, Martin

    2011-09-01

    Night sky brightness is a major source of noise both for Cherenkov telescopes as well as for wide-angle Cherenkov detectors. Therefore, it is important to know the level of night sky brightness at potential sites for future experiments. The measurements of night sky brightness presented here were carried out at Fowler's Gap, a research station in New South Wales, Australia, which is a potential site for the proposed TenTen Cherenkov telescope system and the planned wide-angle Cherenkov detector system HiSCORE. A portable instrument was developed and measurements of the night sky brightness were taken in February and August 2010. Brightness levels were measured for a range of different sky regions and in various spectral bands. The night sky brightness in the relevant wavelength regime for photomultipliers was found to be at the same level as measured in similar campaigns at the established Cherenkov telescope sites of Khomas, Namibia, and at La Palma. The brightness of dark regions in the sky is about 2 × 10 12 photons/(s sr m 2) between 300 nm and 650 nm, and up to four times brighter in bright regions of the sky towards the galactic plane. The brightness in V band is 21.6 magnitudes per arcsec 2 in the dark regions. All brightness levels are averaged over the field of view of the instrument of about 1.3 × 10 -3 sr. The spectrum of the night sky brightness was found to be dominated by longer wavelengths, which allows to apply filters to separate the night sky brightness from the blue Cherenkov light. The possible gain in the signal to noise ratio was found to be up to 1.2, assuming an ideal low-pass filter.

  2. Note: A rectangular pulse generator for 50 kV voltage, 0.8 ns rise time, and 10 ns pulse width based on polymer-film switch.

    PubMed

    Wu, Hanyu; Zhang, Xinjun; Sun, Tieping; Zeng, Zhengzhong; Cong, Peitian; Zhang, Shaoguo

    2015-10-01

    In this article, we describe a rectangular pulse generator, consisting of a polymer-film switch, a tri-plate transmission line, and parallel post-shaped ceramic resistor load, for 50-kV voltage, 0.8-ns rise time, and 10-ns width. The switch and resistors are arranged in atmospheric air and the transmission line can work in atmospheric air or in transformer oil to change the pulse width from 6.7 ns to 10 ns. The fast switching and low-inductance characteristics of the polymer-film switch ensure the fast rising wavefront of <1 ns. This generator can be applied in the calibration of nanosecond voltage dividers and used for electromagnetic pulse tests as a fast-rising current injection source. PMID:26521006

  3. Development of novel antibodies against non-structural proteins nsP1, nsP3 and nsP4 of chikungunya virus: potential use in basic research.

    PubMed

    Kumar, Sameer; Mamidi, Prabhudutta; Kumar, Abhishek; Basantray, Itishree; Bramha, Umarani; Dixit, Anshuman; Maiti, Prasanta Kumar; Singh, Sujay; Suryawanshi, Amol Ratnakar; Chattopadhyay, Subhasis; Chattopadhyay, Soma

    2015-11-01

    Chikungunya virus (CHIKV) has reemerged recently as an important pathogen, causing several large epidemics worldwide. This necessitates the development of better reagents to understand its biology and to establish effective and safe control measures. The present study describes the development and characterization of polyclonal antibodies (pAbs) against synthetic peptides of CHIKV non-structural proteins (nsPs; nsP1, nsP3 and nsP4). The reactivity of these pAbs was demonstrated by ELISA and Western blot. Additionally, in vitro infection studies in a mammalian system confirmed that these pAbs are highly sensitive and specific for CHIKV nsPs, as these proteins were detected very early during viral replication. Homology analysis of the selected epitope sequences revealed that they are conserved among all of the CHIKV strains of different genotypes, while comparison with other alphavirus sequences showed that none of them are 100% identical to the epitope sequences (except Onyong-nyong and Igbo Ora viruses, which show 100% identity to the nsP4 epitope). Interestingly, two different forms of CHIKV nsP1 and three different forms of nsP3 were detected in Western blot analysis during infection; however, further experimental investigations are required to confirm their identity. Also, the use of these antibodies demonstrated faster and enhanced expression profiles of all CHIKV nsPs in 2006 Indian outbreak strains when compared to the CHIKV prototype strain, suggesting the epidemic potential of the 2006 isolate. Accordingly, it can be suggested that the pAbs reported in this study can be used as sensitive and specific tools for experimental investigations of CHIKV replication and infection. PMID:26280524

  4. Development of novel antibodies against non-structural proteins nsP1, nsP3 and nsP4 of chikungunya virus: potential use in basic research.

    PubMed

    Kumar, Sameer; Mamidi, Prabhudutta; Kumar, Abhishek; Basantray, Itishree; Bramha, Umarani; Dixit, Anshuman; Maiti, Prasanta Kumar; Singh, Sujay; Suryawanshi, Amol Ratnakar; Chattopadhyay, Subhasis; Chattopadhyay, Soma

    2015-11-01

    Chikungunya virus (CHIKV) has reemerged recently as an important pathogen, causing several large epidemics worldwide. This necessitates the development of better reagents to understand its biology and to establish effective and safe control measures. The present study describes the development and characterization of polyclonal antibodies (pAbs) against synthetic peptides of CHIKV non-structural proteins (nsPs; nsP1, nsP3 and nsP4). The reactivity of these pAbs was demonstrated by ELISA and Western blot. Additionally, in vitro infection studies in a mammalian system confirmed that these pAbs are highly sensitive and specific for CHIKV nsPs, as these proteins were detected very early during viral replication. Homology analysis of the selected epitope sequences revealed that they are conserved among all of the CHIKV strains of different genotypes, while comparison with other alphavirus sequences showed that none of them are 100% identical to the epitope sequences (except Onyong-nyong and Igbo Ora viruses, which show 100% identity to the nsP4 epitope). Interestingly, two different forms of CHIKV nsP1 and three different forms of nsP3 were detected in Western blot analysis during infection; however, further experimental investigations are required to confirm their identity. Also, the use of these antibodies demonstrated faster and enhanced expression profiles of all CHIKV nsPs in 2006 Indian outbreak strains when compared to the CHIKV prototype strain, suggesting the epidemic potential of the 2006 isolate. Accordingly, it can be suggested that the pAbs reported in this study can be used as sensitive and specific tools for experimental investigations of CHIKV replication and infection.

  5. Investigation of the moving structures in a coronal bright point

    SciTech Connect

    Ning, Zongjun; Guo, Yang

    2014-10-10

    We have explored the moving structures in a coronal bright point (CBP) observed by the Solar Dynamic Observatory Atmospheric Imaging Assembly (AIA) on 2011 March 5. This CBP event has a lifetime of ∼20 minutes and is bright with a curved shape along a magnetic loop connecting a pair of negative and positive fields. AIA imaging observations show that a lot of bright structures are moving intermittently along the loop legs toward the two footpoints from the CBP brightness core. Such moving bright structures are clearly seen at AIA 304 Å. In order to analyze their features, the CBP is cut along the motion direction with a curved slit which is wide enough to cover the bulk of the CBP. After integrating the flux along the slit width, we get the spacetime slices at nine AIA wavelengths. The oblique streaks starting from the edge of the CBP brightness core are identified as moving bright structures, especially on the derivative images of the brightness spacetime slices. They seem to originate from the same position near the loop top. We find that these oblique streaks are bi-directional, simultaneous, symmetrical, and periodic. The average speed is about 380 km s{sup –1}, and the period is typically between 80 and 100 s. Nonlinear force-free field extrapolation shows the possibility that magnetic reconnection takes place during the CBP, and our findings indicate that these moving bright structures could be the observational outflows after magnetic reconnection in the CBP.

  6. Suitable technological conditions for enzymatic hydrolysis of waste paper by Novozymes® enzymes NS50013 and NS50010.

    PubMed

    Brummer, Vladimir; Skryja, Pavel; Jurena, Tomas; Hlavacek, Viliam; Stehlik, Petr

    2014-10-01

    Waste paper belongs to a group of quantitatively the most produced waste types. Enzymatic hydrolysis is becoming a suitable way to treat this type of waste and at the same time, to produce a valuable liquid biofuel, because reducing sugars solutions that are formed during the process of saccharification can be a precursor for following or simultaneous fermentation. If it will be possible to make the enzymatic hydrolysis of the waste paper economically viable, it could serve as one of the new ways to lower the dependence of the transport sector on oil in the future. Only several studies comparing the enzymatic hydrolysis of different waste papers were performed in the past; they are summarized in this manuscript. In our experimental trials, suitable technological conditions for waste paper enzymatic hydrolysis using enzymes from Novozymes® biomass kit: enzymes NS50013 and NS50010 were investigated. The following enzymatic hydrolysis parameters in laboratory scale trials were verified on high cellulose content substrates-filter paper and cellulose pulp: type of buffer, pH, temperature, concentration of the substrate, loading of the enzyme and rate of stirring.

  7. In Silico Screening, Alanine Mutation, and DFT Approaches for Identification of NS2B/NS3 Protease Inhibitors

    PubMed Central

    Balajee, R.; Srinivasadesikan, V.; Sakthivadivel, M.; Gunasekaran, P.

    2016-01-01

    To identify the ligand that binds to a target protein with high affinity is a nontrivial task in computer-assisted approaches. Antiviral drugs have been identified for NS2B/NS3 protease enzyme on the mechanism to cleave the viral protein using the computational tools. The consequence of the molecular docking, free energy calculations, and simulation protocols explores the better ligand. It provides in-depth structural insights with the catalytic triad of His51, Asp75, Ser135, and Gly133. The MD simulation was employed here to predict the stability of the complex. The alanine mutation has been performed and its stability was monitored by using the molecular dynamics simulation. The minimal RMSD value suggests that the derived complexes are close to equilibrium. The DFT outcome reveals that the HOMO-LUMO gap of Ligand19 is 2.86 kcal/mol. Among the considered ligands, Ligand19 shows the lowest gap and it is suggested that the HOMO of Ligand19 may transfer the electrons to the LUMO in the active regions. The calculated binding energy of Ligand19 using the DFT method is in good agreement with the docking studies. The pharmacological activity of ligand was performed and satisfies Lipinski rule of 5. Moreover, the computational results are compared with the available IC50 values of experimental results. PMID:27057355

  8. Characterization of the Determinants of NS2-3-Independent Virion Morphogenesis of Pestiviruses

    PubMed Central

    Klemens, O.; Dubrau, D.

    2015-01-01

    ABSTRACT A peculiarity of the Flaviviridae is the critical function of nonstructural (NS) proteins for virus particle formation. For pestiviruses, like bovine viral diarrhea virus (BVDV), uncleaved NS2-3 represents an essential factor for virion morphogenesis, while NS3 is an essential component of the viral replicase. Accordingly, in natural pestivirus isolates, processing at the NS2-3 cleavage site is not complete, to allow for virion morphogenesis. Virion morphogenesis of the related hepatitis C virus (HCV) shows a major deviation from that of pestiviruses: while RNA replication also requires free NS3, virion formation does not depend on uncleaved NS2-NS3. Recently, we described a BVDV-1 chimera based on strain NCP7 encompassing the NS2-4B*-coding region of strain Osloss (E. Lattwein, O. Klemens, S. Schwindt, P. Becher, and N. Tautz, J Virol 86:427–437, 2012, doi:10.1128/JVI.06133-11). This chimera allowed for the production of infectious virus particles in the absence of uncleaved NS2-3. The Osloss sequence deviates in the NS2-4B* part from NCP7 in 48 amino acids and also has a ubiquitin insertion between NS2 and NS3. The present study demonstrates that in the NCP7 backbone, only two amino acid exchanges in NS2 (E1576V) and NS3 (V1721A) are sufficient and necessary to allow for efficient NS2-3-independent virion morphogenesis. The adaptation of a bicistronic virus encompassing an internal ribosomal entry site element between the NS2 and NS3 coding sequences to efficient virion morphogenesis led to the identification of additional amino acids in E2, NS2, and NS5B that are critically involved in this process. The surprisingly small requirements for approximating the packaging schemes of pestiviruses and HCV with respect to the NS2-3 region is in favor of a common mechanism in an ancestral virus. IMPORTANCE For positive-strand RNA viruses, the processing products of the viral polyprotein serve in RNA replication as well as virion morphogenesis. For bovine viral

  9. High Brightness Neutron Source for Radiography

    SciTech Connect

    Cremer, J. T.; Piestrup, Melvin, A.; Gary, Charles, K.; Harris, Jack, L. Williams, David, J.; Jones, Glenn, E.; Vainionpaa, J. , H.; Fuller, Michael, J.; Rothbart, George, H.; Kwan, J., W.; Ludewigt, B., A.; Gough, R.., A..; Reijonen, Jani; Leung, Ka-Ngo

    2008-12-08

    This research and development program was designed to improve nondestructive evaluation of large mechanical objects by providing both fast and thermal neutron sources for radiography. Neutron radiography permits inspection inside objects that x-rays cannot penetrate and permits imaging of corrosion and cracks in low-density materials. Discovering of fatigue cracks and corrosion in piping without the necessity of insulation removal is possible. Neutron radiography sources can provide for the nondestructive testing interests of commercial and military aircraft, public utilities and petrochemical organizations. Three neutron prototype neutron generators were designed and fabricated based on original research done at the Lawrence Berkeley National Laboratory (LBNL). The research and development of these generators was successfully continued by LBNL and Adelphi Technology Inc. under this STTR. The original design goals of high neutron yield and generator robustness have been achieved, using new technology developed under this grant. In one prototype generator, the fast neutron yield and brightness was roughly 10 times larger than previously marketed neutron generators using the same deuterium-deuterium reaction. In another generator, we integrate a moderator with a fast neutron source, resulting in a high brightness thermal neutron generator. The moderator acts as both conventional moderator and mechanical and electrical support structure for the generator and effectively mimics a nuclear reactor. In addition to the new prototype generators, an entirely new plasma ion source for neutron production was developed. First developed by LBNL, this source uses a spiral antenna to more efficiently couple the RF radiation into the plasma, reducing the required gas pressure so that the generator head can be completely sealed, permitting the possible use of tritium gas. This also permits the generator to use the deuterium-tritium reaction to produce 14-MeV neutrons with increases

  10. The role of the Fraunhofer lines in solar brightness variability

    NASA Astrophysics Data System (ADS)

    Shapiro, A. I.; Solanki, S. K.; Krivova, N. A.; Tagirov, R. V.; Schmutz, W. K.

    2015-09-01

    Context. The solar brightness varies on timescales from minutes to decades. A clear identification of the physical processes behind such variations is needed for developing and improving physics-based models of solar brightness variability and reconstructing solar brightness in the past. This is, in turn, important for better understanding the solar-terrestrial and solar-stellar connections. Aims: We estimate the relative contributions of the continuum, molecular, and atomic lines to the solar brightness variations on different timescales. Methods: Our approach is based on the assumption that variability of the solar brightness on timescales greater than a day is driven by the evolution of the solar surface magnetic field. We calculated the solar brightness variations employing the solar disc area coverage of magnetic features deduced from the MDI/SOHO observations. The brightness contrasts of magnetic features relative to the quiet Sun were calculated with a non-LTE radiative transfer code as functions of disc position and wavelength. By consecutive elimination of molecular and atomic lines from the radiative transfer calculations, we assessed the role of these lines in producing solar brightness variability. Results: We show that the variations in Fraunhofer lines define the amplitude of the solar brightness variability on timescales greater than a day and even the phase of the total solar irradiance variability over the 11-year cycle. We also demonstrate that molecular lines make substantial contribution to solar brightness variability on the 11-year activity cycle and centennial timescales. In particular, our model indicates that roughly a quarter of the total solar irradiance variability over the 11-year cycle originates in molecular lines. The maximum of the absolute spectral brightness variability on timescales greater than a day is associated with the CN violet system between 380 and 390 nm.

  11. Dengue NS1 antigen contributes to disease severity by inducing interleukin (IL)-10 by monocytes.

    PubMed

    Adikari, T N; Gomes, L; Wickramasinghe, N; Salimi, M; Wijesiriwardana, N; Kamaladasa, A; Shyamali, N L A; Ogg, G S; Malavige, G N

    2016-04-01

    Both dengue NS1 antigen and serum interleukin (IL)-10 levels have been shown to associate with severe clinical disease in acute dengue infection, and IL-10 has also been shown to suppress dengue-specific T cell responses. Therefore, we proceeded to investigate the mechanisms by which dengue NS1 contributes to disease pathogenesis and if it is associated with altered IL-10 production. Serum IL-10 and dengue NS1 antigen levels were assessed serially in 36 adult Sri Lankan individuals with acute dengue infection. We found that the serum IL-10 levels correlated positively with dengue NS1 antigen levels (Spearman's r = 0·47, P < 0·0001), and NS1 also correlated with annexin V expression by T cells in acute dengue (Spearman's r = 0·63, P = 0·001). However, NS1 levels did not associate with the functionality of T cell responses or with expression of co-stimulatory molecules. Therefore, we further assessed the effect of dengue NS1 on monocytes and T cells by co-culturing primary monocytes and peripheral blood mononuclear cells (PBMC), with varying concentrations of NS1 for up to 96 h. Monocytes co-cultured with NS1 produced high levels of IL-10, with the highest levels seen at 24 h, and then declined gradually. Therefore, our data show that dengue NS1 appears to contribute to pathogenesis of dengue infection by inducing IL-10 production by monocytes.

  12. Characterising Non-Structural Protein NS4 of African Horse Sickness Virus

    PubMed Central

    Zwart, Lizahn; Potgieter, Christiaan A.; Clift, Sarah J.; van Staden, Vida

    2015-01-01

    African horse sickness is a serious equid disease caused by the orbivirus African horse sickness virus (AHSV). The virus has ten double-stranded RNA genome segments encoding seven structural and three non-structural proteins. Recently, an additional protein was predicted to be encoded by genome segment 9 (Seg-9), which also encodes VP6, of most orbiviruses. This has since been confirmed in bluetongue virus and Great Island virus, and the non-structural protein was named NS4. In this study, in silico analysis of AHSV Seg-9 sequences revealed the existence of two main types of AHSV NS4, designated NS4-I and NS4-II, with different lengths and amino acid sequences. The AHSV NS4 coding sequences were in the +1 reading frame relative to that of VP6. Both types of AHSV NS4 were expressed in cultured mammalian cells, with sizes close to the predicted 17–20 kDa. Fluorescence microscopy of these cells revealed a dual cytoplasmic and nuclear, but not nucleolar, distribution that was very similar for NS4-I and NS4-II. Immunohistochemistry on heart, spleen, and lung tissues from AHSV-infected horses showed that NS4 occurs in microvascular endothelial cells and mononuclear phagocytes in all of these tissues, localising to the both the cytoplasm and the nucleus. Interestingly, NS4 was also detected in stellate-shaped dendritic macrophage-like cells with long cytoplasmic processes in the red pulp of the spleen. Finally, nucleic acid protection assays using bacterially expressed recombinant AHSV NS4 showed that both types of AHSV NS4 bind dsDNA, but not dsRNA. Further studies will be required to determine the exact function of AHSV NS4 during viral replication. PMID:25915516

  13. Characterising Non-Structural Protein NS4 of African Horse Sickness Virus.

    PubMed

    Zwart, Lizahn; Potgieter, Christiaan A; Clift, Sarah J; van Staden, Vida

    2015-01-01

    African horse sickness is a serious equid disease caused by the orbivirus African horse sickness virus (AHSV). The virus has ten double-stranded RNA genome segments encoding seven structural and three non-structural proteins. Recently, an additional protein was predicted to be encoded by genome segment 9 (Seg-9), which also encodes VP6, of most orbiviruses. This has since been confirmed in bluetongue virus and Great Island virus, and the non-structural protein was named NS4. In this study, in silico analysis of AHSV Seg-9 sequences revealed the existence of two main types of AHSV NS4, designated NS4-I and NS4-II, with different lengths and amino acid sequences. The AHSV NS4 coding sequences were in the +1 reading frame relative to that of VP6. Both types of AHSV NS4 were expressed in cultured mammalian cells, with sizes close to the predicted 17-20 kDa. Fluorescence microscopy of these cells revealed a dual cytoplasmic and nuclear, but not nucleolar, distribution that was very similar for NS4-I and NS4-II. Immunohistochemistry on heart, spleen, and lung tissues from AHSV-infected horses showed that NS4 occurs in microvascular endothelial cells and mononuclear phagocytes in all of these tissues, localising to the both the cytoplasm and the nucleus. Interestingly, NS4 was also detected in stellate-shaped dendritic macrophage-like cells with long cytoplasmic processes in the red pulp of the spleen. Finally, nucleic acid protection assays using bacterially expressed recombinant AHSV NS4 showed that both types of AHSV NS4 bind dsDNA, but not dsRNA. Further studies will be required to determine the exact function of AHSV NS4 during viral replication.

  14. Discovery of a bright eclipsing cataclysmic variable

    NASA Astrophysics Data System (ADS)

    Sing, D. K.; Green, E. M.; Howell, S. B.; Holberg, J. B.; Lopez-Morales, M.; Shaw, J. S.; Schmidt, G. D.

    2007-11-01

    Aims:We report on the discovery of J0644+3344, a bright, deeply-eclipsing cataclysmic variable (CV) binary. Methods: Optical photometric and spectroscopic observations were obtained to determine the nature and characteristics of this CV. Results: Spectral signatures of both binary components and an accretion disk can be seen at optical wavelengths. The optical spectrum shows broad H I, He I, and He II accretion disk emission lines with deep narrow absorption components from H I, He I, Mg II, and Ca II. The absorption lines are seen throughout the orbital period, disappearing only during primary eclipse. These absorption lines are either the result of an optically-thick inner accretion disk or from the photosphere of the primary star. Radial velocity measurements show that the H I, He I, and Mg II absorption lines phase with the primary star, while weak absorption features in the continuum, between Hα and Hβ, phase with the secondary star. Radial velocity solutions give a 150±4 km s-1 semi-amplitude for the primary star and 192.8±5.6 km s-1 for the secondary, resulting in a primary to secondary mass ratio of q = 1.285. The individual stellar masses are 0.63-0.69 M⊙ for the primary and 0.49-0.54 M⊙ for the secondary, with the uncertainty largely due to the inclination. Conclusions: The bright eclipsing nature of this binary has helped provide masses for both components with an accuracy rarely achieved for CVs. This binary most closely resembles a nova-like UX UMa or SW Sex type of CV. J0644+3344, however, has a longer orbital period than most UX UMa or SW Sex stars. Assuming an evolution toward shorter orbital periods, J0644+3344 is therefore likely to be a young interacting binary. The secondary star is consistent with the size and spectral type of a K8 star, but has the mass of a M0.

  15. The 150 ns detector project: Progress with small detectors

    NASA Astrophysics Data System (ADS)

    Warburton, W. K.; Russell, S. R.; Kleinfelder, Stuart A.; Segal, Julie

    1994-09-01

    This project's long term goal is to develop a pixel area detector capable of 6 MHz frame rates (150 ns/frame). Our milestones toward this goal are: a single pixel, 1 × 256 1D and 8 × 8 2D detectors, 256 × 256 2D detectors and, finally, 1024 × 1024 2D detectors. The design strategy is to supply a complete electronics chain (resetting preamp, selectable gain amplifier, analog-to-digital converter (ADC), and memory) for each pixel. In the final detectors these will all be custom integrated circuits. The front end preamplifiers are being integrated first, since their design and performance are both the most unusual and also critical to the project's success. Similarly, our early work is also concentrating on devising and perfecting detector structures which are thick enough (1 mm) to absorb over 99% of the incident X-rays in the energy range of interest. In this paper we discuss our progress toward the 1 × 256 1D and 8 × 8 2D detectors. We have fabricated sample detectors at Stanford's Center for Integrated Systems and are preparing both to test them individually and to wirebond them to the preamplifier samples to produce our first working small 1D and 2D detectors. We will describe our solutions to the design problems associated with collecting charge in less than 30 ns from 1 mm thick pixels in high resistivity silicon. We have constructed and tested the front end of our preamplifier design using a commercial 1.2 μm CMOS technology and are moving on to produce a few channels of the complete preamplifier, including a switchable gain stage and output stage. We will discuss both the preamplifier design and our initial test results.

  16. Spain 31-GHz observations of sky brightness temperatures

    NASA Technical Reports Server (NTRS)

    Gary, B. L.

    1988-01-01

    A water vapor radiometer was deployed at DSS 63 for 3 months of sky brightness temperature measurements at 31 GHz. An exceedance plot was derived from this data showing the fraction of time that 31 GHz 30 degree elevation angle brightness temperature exceeds specified values. The 5 percent exceedance statistics occurs at 75 K, compared with 70 K in Australia.

  17. Analysis of Bright Harvest Remote Analysis for Residential Solar Installations

    SciTech Connect

    Nangle, John; Simon, Joseph

    2015-06-17

    Bright Harvest provides remote shading analysis and design products for residential PV system installers. The National Renewable Energy Laboratory (NREL) through the NREL Commercialization Assistance Program, completed comparative assessments between on-site measurements and remotely calculated values to validate the accuracy of Bright Harvest’s remote shading and power generation.

  18. 7 CFR 51.2000 - Clean and bright.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 Agriculture 2 2012-01-01 2012-01-01 false Clean and bright. 51.2000 Section 51.2000 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Standards for Grades of Filberts in the Shell 1 Definitions § 51.2000 Clean and bright. Clean and...

  19. 7 CFR 51.2000 - Clean and bright.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Clean and bright. 51.2000 Section 51.2000 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Standards for Grades of Filberts in the Shell 1 Definitions § 51.2000 Clean and bright. Clean and...

  20. 7 CFR 51.2000 - Clean and bright.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Clean and bright. 51.2000 Section 51.2000 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing... Standards for Grades of Filberts in the Shell 1 Definitions § 51.2000 Clean and bright. Clean and...

  1. Challenging Exceptionally Bright Children in Early Childhood Classrooms

    ERIC Educational Resources Information Center

    Gadzikowski, Ann

    2013-01-01

    Nearly every group of children includes at least one exceptionally bright child. From the especially creative child to the child who has already mastered learning outcomes to the "twice exceptional" child, exceptionally bright children have a wide range of talents and behaviors. This book will help you understand what it means to be…

  2. Bright visible light emission from graphene

    NASA Astrophysics Data System (ADS)

    Kim, Young Duck; Kim, Hakseong; Cho, Yujin; Ryoo, Ji Hoon; Park, Cheol-Hwan; Kim, Pilkwang; Kim, Yong Seung; Lee, Sunwoo; Li, Yilei; Park, Seung-Nam; Shim Yoo, Yong; Yoon, Duhee; Dorgan, Vincent E.; Pop, Eric; Heinz, Tony F.; Hone, James; Chun, Seung-Hyun; Cheong, Hyeonsik; Lee, Sang Wook; Bae, Myung-Ho; Park, Yun Daniel

    2015-08-01

    Graphene and related two-dimensional materials are promising candidates for atomically thin, flexible and transparent optoelectronics. In particular, the strong light-matter interaction in graphene has allowed for the development of state-of-the-art photodetectors, optical modulators and plasmonic devices. In addition, electrically biased graphene on SiO2 substrates can be used as a low-efficiency emitter in the mid-infrared range. However, emission in the visible range has remained elusive. Here, we report the observation of bright visible light emission from electrically biased suspended graphene devices. In these devices, heat transport is greatly reduced. Hot electrons (˜2,800 K) therefore become spatially localized at the centre of the graphene layer, resulting in a 1,000-fold enhancement in thermal radiation efficiency. Moreover, strong optical interference between the suspended graphene and substrate can be used to tune the emission spectrum. We also demonstrate the scalability of this technique by realizing arrays of chemical-vapour-deposited graphene light emitters. These results pave the way towards the realization of commercially viable large-scale, atomically thin, flexible and transparent light emitters and displays with low operation voltage and graphene-based on-chip ultrafast optical communications.

  3. High output lamp with high brightness

    DOEpatents

    Kirkpatrick, Douglas A.; Bass, Gary K.; Copsey, Jesse F.; Garber, Jr., William E.; Kwong, Vincent H.; Levin, Izrail; MacLennan, Donald A.; Roy, Robert J.; Steiner, Paul E.; Tsai, Peter; Turner, Brian P.

    2002-01-01

    An ultra bright, low wattage inductively coupled electrodeless aperture lamp is powered by a solid state RF source in the range of several tens to several hundreds of watts at various frequencies in the range of 400 to 900 MHz. Numerous novel lamp circuits and components are disclosed including a wedding ring shaped coil having one axial and one radial lead, a high accuracy capacitor stack, a high thermal conductivity aperture cup and various other aperture bulb configurations, a coaxial capacitor arrangement, and an integrated coil and capacitor assembly. Numerous novel RF circuits are also disclosed including a high power oscillator circuit with reduced complexity resonant pole configuration, parallel RF power FET transistors with soft gate switching, a continuously variable frequency tuning circuit, a six port directional coupler, an impedance switching RF source, and an RF source with controlled frequency-load characteristics. Numerous novel RF control methods are disclosed including controlled adjustment of the operating frequency to find a resonant frequency and reduce reflected RF power, controlled switching of an impedance switched lamp system, active power control and active gate bias control.

  4. The Los Alamos high-brightness photoinjector

    SciTech Connect

    O'Shea, P.G.

    1991-01-01

    For a number of years Los Alamos National Laboratory has been developing photocathode RF guns for high-brightness electron beam applications such as free-electron lasers (FELs). Previously thermionic high-voltage guns have been the source of choice for the electron accelerators used to drive FELs. The performance of such FELs is severely limited by the emittance growth produced by the subharmonic bunching process and also by the low peak current of the source. In a photoinjector, a laser driven photocathode is placed directly in a high-gradient RF accelerating cavity. A photocathode allows unsurpassed control over the current, and the spatial and temporal profile of the beam. In addition the electrodeless emission'' avoids many of the difficulties associated with multi-electrode guns, i.e. the electrons are accelerated very rapidly to relativistic energies, and there are no electrodes to distort the accelerating fields. For the past two years we have been integrating a photocathode into our existing FEL facility by replacing our thermionic gun and subharmonic bunchers with a high-gradient 1.3 GHz photoinjector. The photoinjector, which is approximately 0.6 m in length, produces 6 MeV, 300 A, 15 ps linac, and accelerated to a final energy of 40 MeV. We have recently begun lasing at wavelengths near 3 {mu}m. 16 refs., 2 figs., 5 tabs.

  5. Lyman Alpha Mapping Project (LAMP) Brightness Maps

    NASA Astrophysics Data System (ADS)

    Retherford, Kurt D.; Gladstone, G.; Stern, S.; Egan, A. F.; Miles, P. F.; Parker, J. W.; Greathouse, T. K.; Davis, M. W.; Slater, D. C.; Kaufmann, D. E.; Versteeg, M. H.; Feldman, P. D.; Hurley, D. M.; Pryor, W. R.; Hendrix, A. R.

    2010-10-01

    The Lyman Alpha Mapping Project (LAMP) is an ultraviolet (UV) spectrograph on the Lunar Reconnaissance Orbiter (LRO) that is designed to map the lunar albedo at far-UV wavelengths. LAMP primarily measures interplanetary Hydrogen Lyman-alpha sky-glow and far-UV starlight reflected from the night-side lunar surface, including permanently shadowed regions (PSRs) near the poles. Dayside observations are also obtained. Brightness maps sorted by wavelength (including the Lyman-alpha wavelength of 121.6 nm) are reported for the polar regions, with a few regions of interest reported in more detail. LAMP's spectral range of 58 nm to 196 nm includes a water ice spectral feature near 160 nm, which provides a diagnostic tool for detecting water on the lunar surface that is complementary to recent discoveries using infrared and radio frequency techniques. Progress towards producing far-UV albedo maps and searching for water ice signatures will be reported. We'll discuss how LAMP data may address questions regarding how water is formed on the moon, transported through the lunar atmosphere, and deposited in the PSRs.

  6. MAGNETIC BRIGHT POINTS IN THE QUIET SUN

    SciTech Connect

    Sanchez Almeida, J.; Bonet, J. A.; Viticchie, B.

    2010-05-20

    We present a visual determination of the number of bright points (BPs) existing in the quiet Sun, which are structures though to trace intense kG magnetic concentrations. The measurement is based on a 0.''1 angular resolution G-band movie obtained with the Swedish Solar Telescope at the solar disk center. We find 0.97 BPs Mm{sup -2}, which is a factor 3 larger than any previous estimate. It corresponds to 1.2 BPs per solar granule. Depending on the details of the segmentation, the BPs cover between 0.9% and 2.2% of the solar surface. Assuming their field strength to be 1.5 kG, the detected BPs contribute to the solar magnetic flux with an unsigned flux density between 13 G and 33 G. If network and inter-network regions are counted separately, they contain 2.2 BPs Mm{sup -2} and 0.85 BPs Mm{sup -2}, respectively.

  7. Dark Skies, Bright Kids Year 6

    NASA Astrophysics Data System (ADS)

    Liss, Sandra; Troup, Nicholas William; Johnson, Kelsey E.; Barcos-Munoz, Loreto D.; Beaton, Rachael; Bittle, Lauren; Borish, Henry J.; Burkhardt, Andrew; Corby, Joanna; Dean, Janice; Hancock, Danielle; King, Jennie; Prager, Brian; Romero, Charles; Sokal, Kimberly R.; Stierwalt, Sabrina; Wenger, Trey; Zucker, Catherine

    2015-01-01

    Now entering our sixth year of operation, Dark Skies, Bright Kids (DSBK) is an entirely volunteer-run outreach organization based out of the Department of Astronomy at the University of Virginia. Our core mission is to enhance elementary science education and literacy in central Virginia through fun, hands-on activities that introduce basic Astronomy concepts beyond Virginia's Standards of Learning. Our primary focus is hosting an 8-10 week after-school astronomy club at underserved elementary and middle schools. Each week, DSBK volunteers take the role of coaches to introduce astronomy-related concepts ranging from the Solar System to galaxies to astrobiology, and to lead students in interactive learning activities. Another hallmark of DSBK is hosting our Annual Central Virginia Star Party, a free event open to the community featuring star-gazing and planetarium shows.DSBK has amassed over 15,000 contact hours since 2009 and we continue to broaden our impact. One important step we have taken in the past year is to establish a graduate student led assessment program to identify and implement directed learning goals for DSBK outreach. The collection of student workbooks, observations, and volunteer surveys indicates broad scale success for the program both in terms of student learning and their perception of science. The data also reveal opportunities to improve our organizational and educational practices to maximize student achievement and overall volunteer satisfaction for DSBK's future clubs and outreach endeavors.

  8. Intercomparisons of nine sky brightness detectors.

    PubMed

    den Outer, Peter; Lolkema, Dorien; Haaima, Marty; van der Hoff, Rene; Spoelstra, Henk; Schmidt, Wim

    2011-01-01

    Nine Sky Quality Meters (SQMs) have been intercompared during a night time measurement campaign held in the Netherlands in April 2011. Since then the nine SQMs have been distributed across The Netherlands and form the Dutch network for monitoring night sky brightness. The goal of the intercomparison was to infer mutual calibration factors and obtain insight into the variability of the SQMs under different meteorological situations. An ensemble average is built from the individual measurements and used as a reference to infer the mutual calibration factors. Data required additional synchronization prior to the calibration determination, because the effect of moving clouds combined with small misalignments emerges as time jitter in the measurements. Initial scatter of the individual instruments lies between ±14%. Individual night time sums range from -16% to +20%. Intercalibration reduces this to 0.5%, and -7% to +9%, respectively. During the campaign the smallest luminance measured was 0.657 ± 0.003 mcd/m(2) on 12 April, and the largest value was 5.94 ± 0.03 mcd/m(2) on 2 April. During both occurrences interfering circumstances like snow cover or moonlight were absent.

  9. Antilensing: the bright side of voids.

    PubMed

    Bolejko, Krzysztof; Clarkson, Chris; Maartens, Roy; Bacon, David; Meures, Nikolai; Beynon, Emma

    2013-01-11

    More than half of the volume of our Universe is occupied by cosmic voids. The lensing magnification effect from those underdense regions is generally thought to give a small dimming contribution: objects on the far side of a void are supposed to be observed as slightly smaller than if the void were not there, which together with conservation of surface brightness implies net reduction in photons received. This is predicted by the usual weak lensing integral of the density contrast along the line of sight. We show that this standard effect is swamped at low redshifts by a relativistic Doppler term that is typically neglected. Contrary to the usual expectation, objects on the far side of a void are brighter than they would be otherwise. Thus the local dynamics of matter in and near the void is crucial and is only captured by the full relativistic lensing convergence. There are also significant nonlinear corrections to the relativistic linear theory, which we show actually underpredicts the effect. We use exact solutions to estimate that these can be more than 20% for deep voids. This remains an important source of systematic errors for weak lensing density reconstruction in galaxy surveys and for supernovae observations, and may be the cause of the reported extra scatter of field supernovae located on the edge of voids compared to those in clusters. PMID:23383886

  10. Antilensing: The Bright Side of Voids

    NASA Astrophysics Data System (ADS)

    Bolejko, Krzysztof; Clarkson, Chris; Maartens, Roy; Bacon, David; Meures, Nikolai; Beynon, Emma

    2013-01-01

    More than half of the volume of our Universe is occupied by cosmic voids. The lensing magnification effect from those underdense regions is generally thought to give a small dimming contribution: objects on the far side of a void are supposed to be observed as slightly smaller than if the void were not there, which together with conservation of surface brightness implies net reduction in photons received. This is predicted by the usual weak lensing integral of the density contrast along the line of sight. We show that this standard effect is swamped at low redshifts by a relativistic Doppler term that is typically neglected. Contrary to the usual expectation, objects on the far side of a void are brighter than they would be otherwise. Thus the local dynamics of matter in and near the void is crucial and is only captured by the full relativistic lensing convergence. There are also significant nonlinear corrections to the relativistic linear theory, which we show actually underpredicts the effect. We use exact solutions to estimate that these can be more than 20% for deep voids. This remains an important source of systematic errors for weak lensing density reconstruction in galaxy surveys and for supernovae observations, and may be the cause of the reported extra scatter of field supernovae located on the edge of voids compared to those in clusters.

  11. Bright visible light emission from graphene.

    PubMed

    Kim, Young Duck; Kim, Hakseong; Cho, Yujin; Ryoo, Ji Hoon; Park, Cheol-Hwan; Kim, Pilkwang; Kim, Yong Seung; Lee, Sunwoo; Li, Yilei; Park, Seung-Nam; Yoo, Yong Shim; Yoon, Duhee; Dorgan, Vincent E; Pop, Eric; Heinz, Tony F; Hone, James; Chun, Seung-Hyun; Cheong, Hyeonsik; Lee, Sang Wook; Bae, Myung-Ho; Park, Yun Daniel

    2015-08-01

    Graphene and related two-dimensional materials are promising candidates for atomically thin, flexible and transparent optoelectronics. In particular, the strong light-matter interaction in graphene has allowed for the development of state-of-the-art photodetectors, optical modulators and plasmonic devices. In addition, electrically biased graphene on SiO2 substrates can be used as a low-efficiency emitter in the mid-infrared range. However, emission in the visible range has remained elusive. Here, we report the observation of bright visible light emission from electrically biased suspended graphene devices. In these devices, heat transport is greatly reduced. Hot electrons (∼2,800 K) therefore become spatially localized at the centre of the graphene layer, resulting in a 1,000-fold enhancement in thermal radiation efficiency. Moreover, strong optical interference between the suspended graphene and substrate can be used to tune the emission spectrum. We also demonstrate the scalability of this technique by realizing arrays of chemical-vapour-deposited graphene light emitters. These results pave the way towards the realization of commercially viable large-scale, atomically thin, flexible and transparent light emitters and displays with low operation voltage and graphene-based on-chip ultrafast optical communications.

  12. A Magnetic Bright Point Case Study

    NASA Astrophysics Data System (ADS)

    Utz, D.; Jurčák, J.; Bellot-Rubio, L.; del Toro Iniesta, J. C.; Thonhofer, S.; Hanslmeier, A.; Veronig, A.; Muller, R.; Lemmerer, B.

    Due to its magnetic fields our host star - the Sun - becomes the interesting object for research as we know it. The magnetic fields themselves cover different spatial, lifetime and strength scales and reach down from enormous flux concentrations like active sunspot groups to single isolated magnetic flux tubes and even weaker, predominantly inclined intranetwork structures. Flux tubes can be seen in filtergram observations as magnetic bright points (MBPs). They are of interest for research not only due to their sheer existence but due to their important role in atmospheric heating (wave heating as well as reconnection processes), to their role in the understanding of creation and annihilation of magnetic fields as well as to their influence on the total solar irradiance variation. In this study we present a close look onto an evolutionary track of an MBP from its formation to its disintegration. Physical quantities of MBPs like their magnetic field strength and inclination, their line-of-sight velocity, and their temperature at different heights are inferred from the inversion of spectropolarimetric data. Original data are taken from the Sunrise/IMaX instrument and constitute a time series of some 60 min. The presented case resembles the convective collapse model and is in agreement with previous studies.

  13. Dengue Virus NS1 Protein Modulates Cellular Energy Metabolism by Increasing Glyceraldehyde-3-Phosphate Dehydrogenase Activity

    PubMed Central

    Allonso, Diego; Andrade, Iamara S.; Conde, Jonas N.; Coelho, Diego R.; Rocha, Daniele C. P.; da Silva, Manuela L.; Ventura, Gustavo T.

    2015-01-01

    ABSTRACT Dengue is one of the main public health concerns worldwide. Recent estimates indicate that over 390 million people are infected annually with the dengue virus (DENV), resulting in thousands of deaths. Among the DENV nonstructural proteins, the NS1 protein is the only one whose function during replication is still unknown. NS1 is a 46- to 55-kDa glycoprotein commonly found as both a membrane-associated homodimer and a soluble hexameric barrel-shaped lipoprotein. Despite its role in the pathogenic process, NS1 is essential for proper RNA accumulation and virus production. In the present study, we identified that glyceraldehyde-3-phosphate dehydrogenase (GAPDH) interacts with intracellular NS1. Molecular docking revealed that this interaction occurs through the hydrophobic protrusion of NS1 and the hydrophobic residues located at the opposite side of the catalytic site. Moreover, addition of purified recombinant NS1 enhanced the glycolytic activity of GAPDH in vitro. Interestingly, we observed that DENV infection promoted the relocalization of GAPDH to the perinuclear region, where NS1 is commonly found. Both DENV infection and expression of NS1 itself resulted in increased GAPDH activity. Our findings indicate that the NS1 protein acts to increase glycolytic flux and, consequently, energy production, which is consistent with the recent finding that DENV induces and requires glycolysis for proper replication. This is the first report to propose that NS1 is an important modulator of cellular energy metabolism. The data presented here provide new insights that may be useful for further drug design and the development of alternative antiviral therapies against DENV. IMPORTANCE Dengue represents a serious public health problem worldwide and is caused by infection with dengue virus (DENV). Estimates indicate that half of the global population is at risk of infection, with almost 400 million cases occurring per year. The NS1 glycoprotein is found in both the

  14. Interpreting Central Surface Brightness and Color Profiles in Elliptical Galaxies

    NASA Astrophysics Data System (ADS)

    Silva, David R.; Wise, Michael W.

    1996-01-01

    Hubble Space Telescope imagery has revealed dust features in the central regions of many (50%--80%) nearby bright elliptical galaxies. If these features are an indication of an underlying smooth diffuse dust distribution, then the interpretation of central surface brightness and color profiles in elliptical galaxies becomes significantly more difficult. In this Letter, diagnostics for constraining the presence of such an underlying central dust distribution are presented. We show that easily detectable central color gradients and flattened central surface brightness profiles can be induced by even small amounts of smoothly distributed dust (~100 M⊙). Conversely, combinations of flat surface brightness profiles and flat color gradients or steep surface brightness profiles and steep color gradients are unlikely to be caused by dust. Taken as a whole, these results provide a simple observational tautology for constraining the existence of smooth diffuse dust distributions in the central regions of elliptical galaxies.

  15. Retinal mesopic adaptation model for brightness perception under transient glare.

    PubMed

    Barrionuevo, Pablo Alejandro; Colombo, Elisa Margarita; Issolio, Luis Alberto

    2013-06-01

    A glare source in the visual field modifies the brightness of a test patch surrounded by a mesopic background. In this study, we investigated the effect of two levels of transient glare on brightness perception for several combinations of mesopic reference test luminances (Lts) and background luminances (Lbs). While brightness perception was affected by Lb, there were no appreciable effects for changes in the Lt. The highest brightness reduction was found for Lbs in the low mesopic range. Considering the main proposal that brightness can be inferred from contrast and the Lb sets the mesopic luminance adaptation, we hypothesized that contrast gain and retinal adaptation mechanisms would act when a transient glare source was present in the visual field. A physiology-based model that adequately fitted the present and previous results was developed.

  16. Brightness perception of unrelated self-luminous colors.

    PubMed

    Withouck, Martijn; Smet, Kevin A G; Ryckaert, Wouter R; Pointer, Michael R; Deconinck, Geert; Koenderink, Jan; Hanselaer, Peter

    2013-06-01

    The perception of brightness of unrelated self-luminous colored stimuli of the same luminance has been investigated. The Helmholtz-Kohlrausch (H-K) effect, i.e., an increase in brightness perception due to an increase in saturation, is clearly observed. This brightness perception is compared with the calculated brightness according to six existing vision models, color appearance models, and models based on the concept of equivalent luminance. Although these models included the H-K effect and half of them were developed to work with unrelated colors, none of the models seemed to be able to fully predict the perceived brightness. A tentative solution to increase the prediction accuracy of the color appearance model CAM97u, developed by Hunt, is presented.

  17. Policresulen, a novel NS2B/NS3 protease inhibitor, effectively inhibits the replication of DENV2 virus in BHK-21 cells

    PubMed Central

    Wu, Deng-wei; Mao, Fei; Ye, Yan; Li, Jian; Xu, Chuan-lian; Luo, Xiao-min; Chen, Jing; Shen, Xu

    2015-01-01

    Aim: Dengue is a severe epidemic disease caused by dengue virus (DENV) infection, for which no effective treatment is available. The protease complex, consisting of nonstructural protein 3 (NS3) and its cofactor NS2B, plays a pivotal role in the replication of DENV, thus may be a potential target for anti-DENV drugs. Here, we report a novel inhibitor of DENV2 NS2B/NS3 protease and its antiviral action. Methods: An enzymatic inhibition assay was used for screening DENV2 NS2B/NS3 inhibitors. Cytotoxicity to BHK-21 cells was assessed with MTT assay. Antiviral activity was evaluated in BHK-21 cells transfected with Rlu-DENV-Rep. The molecular mechanisms of the antiviral action was analyzed using surface plasmon resonance, ultraviolet-visible spectral analysis and differential scanning calorimetry assays, as well as molecular docking analysis combined with site-directed mutagenesis. Results: In our in-house library of old drugs (∼1000 compounds), a topical hemostatic and antiseptic 2-hydroxy-3,5-bis[(4-hydroxy-2-methyl-5-sulfophenyl)methyl]-4-methyl-benzene-sulfonic acid (policresulen) was found to be a potent inhibitor of DENV2 NS2B/NS3 protease with IC50 of 0.48 μg/mL. Furthermore, policresulen inhibited DENV2 replication in BHK-21 cells with IC50 of 4.99 μg/mL, whereas its IC50 for cytotoxicity to BHK-21 cells was 459.45 μg/mL. Policresulen acted as a competitive inhibitor of the protease, and slightly affected the protease stability. Using biophysical technology-based assays and molecular docking analysis combined with site-directed mutagenesis, we demonstrated that the residues Gln106 and Arg133 of DENV2 NS2B/NS3 protease directly interacted with policresulen via hydrogen bonding. Conclusion: Policresulen is a potent inhibitor of DENV2 NS2B/NS3 protease that inhibits DENV2 replication in BHK-21 cells. The binding mode of the protease and policresulen provides useful hints for designing new type of inhibitors against the protease. PMID:26279156

  18. VP2 Exchange and NS3/NS3a Deletion in African Horse Sickness Virus (AHSV) in Development of Disabled Infectious Single Animal Vaccine Candidates for AHSV

    PubMed Central

    van de Water, Sandra G. P.; van Gennip, René G. P.; Potgieter, Christiaan A.; Wright, Isabel M.

    2015-01-01

    ABSTRACT African horse sickness virus (AHSV) is a virus species in the genus Orbivirus of the family Reoviridae. There are nine serotypes of AHSV showing different levels of cross neutralization. AHSV is transmitted by species of Culicoides biting midges and causes African horse sickness (AHS) in equids, with a mortality rate of up to 95% in naive horses. AHS has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate climates appear to be competent vectors for the related bluetongue virus (BTV). To control AHS, live-attenuated vaccines (LAVs) are used in Africa. We used reverse genetics to generate “synthetic” reassortants of AHSV for all nine serotypes by exchange of genome segment 2 (Seg-2). This segment encodes VP2, which is the serotype-determining protein and the dominant target for neutralizing antibodies. Single Seg-2 AHSV reassortants showed similar cytopathogenic effects in mammalian cells but displayed different growth kinetics. Reverse genetics for AHSV was also used to study Seg-10 expressing NS3/NS3a proteins. We demonstrated that NS3/NS3a proteins are not essential for AHSV replication in vitro. NS3/NS3a of AHSV is, however, involved in the cytopathogenic effect in mammalian cells and is very important for virus release from cultured insect cells in particular. Similar to the concept of the bluetongue disabled infectious single animal (BT DISA) vaccine platform, an AHS DISA vaccine platform lacking NS3/NS3a expression was developed. Using exchange of genome segment 2 encoding VP2 protein (Seg-2[VP2]), we will be able to develop AHS DISA vaccine candidates for all current AHSV serotypes. IMPORTANCE African horse sickness virus is transmitted by species of Culicoides biting midges and causes African horse sickness in equids, with a mortality rate of up to 95% in naive horses. African horse sickness has become a serious threat for countries outside Africa, since endemic Culicoides species in moderate

  19. [Grouping of the NS1 nonstructural proteins of influenza A viruses].

    PubMed

    Sokolov, B P; Rudneva, I A; Zhdanov, V M

    1981-01-01

    Peptide mapping was used for comparative analysis of nonstructural proteins (NS1) of 21 strains of human and animal influenza A viruses. At least 4 groups of NS1 proteins could be distinguished by the analysis of the peptide maps; we designated these groups as 0, 1, 2, and 3. Group O includes NS1 proteins of human influenza virus serotype HON1, group 1 - NS1 proteins of viruses of serotypes H1N1 and H2N2, group 2 - NS1 proteins of viruses of serotype H3N2. NS1 proteins of avian influenza viruses A/duck Czechoslovakia/63, A/turkey Massachusetts/65, A/petrel Australia/1/71, A/duck Ukraine/63, and A/turkey Ontario/68 have been included into group 3. PMID:7336689

  20. Functions of the influenza A virus NS1 protein in antiviral defense.

    PubMed

    Krug, Robert M

    2015-06-01

    Influenza A viruses counteract host antiviral activities, especially the production of interferons (IFNs) and the activities of IFN-induced proteins that inhibit virus replication. The viral NS1 protein is largely responsible for countering these IFN antiviral responses, but there are functional differences between the NS1 proteins of different virus strains. The NS1 protein inhibits IFN production by two mechanisms: inhibition of the activation of IRF3 and IFN transcription; and inhibition of the processing of IFN pre-mRNAs. The NS1 proteins of several virus strains do not inhibit IRF3 activation, and the NS1 protein of one virus strain does not inhibit the processing of IFN pre-mRNAs. Many issues remain concerning the mechanisms of action of the various NS1 proteins in countering the IFN response.

  1. Functions of the Influenza A Virus NS1 Protein In Antiviral Defense

    PubMed Central

    Krug, Robert M.

    2015-01-01

    Influenza A viruses counteract host antiviral activities, especially the production of interferons (IFNs) and the activities of IFN-induced proteins that inhibit virus replication. The viral NS1 protein is largely responsible for countering these IFN antiviral responses, but there are functional differences between the NS1 proteins of different virus strains. The NS1 protein inhibits IFN production by two mechanisms: inhibition of the activation of IRF3 and IFN transcription; and inhibition of the processing of IFN pre-mRNAs. The NS1 proteins of several virus strains do not inhibit IRF3 activation, and the NS1 protein of one virus strain does not inhibit the processing of IFN pre-mRNAs. Many issues remain concerning the mechanisms of action of the various NS1 proteins in countering the IFN response. PMID:25638592

  2. Structure of NS1A effector domain from the influenza A/Udorn/72 virus

    SciTech Connect

    Xia, Shuangluo; Monzingo, Arthur F.; Robertus, Jon D.

    2009-01-01

    The structure of the effector domain of the influenza protein NS1, a validated antiviral drug target, has been solved in two space groups. The nonstructural protein NS1A from influenza virus is a multifunctional virulence factor and a potent inhibitor of host immunity. It has two functional domains: an N-terminal 73-amino-acid RNA-binding domain and a C-terminal effector domain. Here, the crystallographic structure of the NS1A effector domain of influenza A/Udorn/72 virus is presented. Structure comparison with the NS1 effector domain from mouse-adapted influenza A/Puerto Rico/8/34 (PR8) virus strain reveals a similar monomer conformation but a different dimer interface. Further analysis and evaluation shows that the dimer interface observed in the structure of the PR8 NS1 effector domain is likely to be a crystallographic packing effect. A hypothetical model of the intact NS1 dimer is presented.

  3. Characterization of the magnetization reversal of perpendicular Nanomagnetic Logic clocked in the ns-range

    NASA Astrophysics Data System (ADS)

    Ziemys, Grazvydas; Trummer, Christian; Gamm, Stephan Breitkreutz-v.; Eichwald, Irina; Schmitt-Landsiedel, Doris; Becherer, Markus

    2016-05-01

    We have investigated the magnetization reversal of fabricated Co/Pt nanomagnets with perpendicular anisotropy within a wide range of magnetic field pulse widths. This experiment covers the pulse lengths from 700 ms to 20 ns. We observed that the commonly used Arrhenius model fits very well the experimental data with a single parameter set for pulse times above 100 ns (tp > 100 ns). However, below 100 ns (tp < 100 ns), a steep increase of the switching field amplitude is observed and the deviation from the Arrhenius model becomes unacceptable. For short pulse times the model can be adjusted by the reversal time term for the dynamic switching field which is only dependent on the pulse amplitude and not on temperature anymore. Precise modeling of the magnetization reversal in the sub-100 ns-range is crucially important to ensure reliable operation in the favored GHz-range as well as to explore and design new kinds of Nanomagnetic Logic circuits and architectures.

  4. Functional differences in hepatitis C virus nonstructural (NS) 3/4A- and 5A-specific T cell responses

    PubMed Central

    Holmström, Fredrik; Chen, Margaret; Balasiddaiah, Anangi; Sällberg, Matti; Ahlén, Gustaf; Frelin, Lars

    2016-01-01

    The hepatitis C virus nonstructural (NS) 3/4A and NS5A proteins are major targets for the new direct-acting antiviral compounds. Both viral proteins have been suggested as modulators of the response to the host cell. We have shown that NS3/4A- and NS5A-specific T cell receptors confer different effector functions, and that killing of NS3/4A-expressing hepatocytes is highly dependent on IFN-γ. We here characterize the functional differences in the T cell responses to NS3/4A and NS5A. NS3/4A- and NS5A-specific T cells could be induced at various frequencies in wild-type-, NS3/4A-, and NS5A-transgenic mice. Priming of NS5A-specific T cells required a high DNA dose, and was unlike NS3/4A dependent on both CD4+ and CD8+ T cells, but less influenced by CD25+/GITR+ regulatory T cells. The presence of IL-12 greatly improved specific CD8+ T cell priming by NS3/4A but not by NS5A, suggesting a less dependence of IFN-γ for NS5A. This notion was supported by the observation that NS5A-specific T cells could eliminate NS5A-expressing hepatocytes also in the absence of IFN-γ-receptor-2. This supports that NS3/4A- and NS5A-specific T cells become activated and eliminate antigen expressing, or infected hepatocytes, by distinct mechanisms, and that NS5A-specific T cells show an overall less dependence of IFN-γ. PMID:27141891

  5. The NS5A protein of hepatitis C virus is a zinc metalloprotein.

    PubMed

    Tellinghuisen, Timothy L; Marcotrigiano, Joseph; Gorbalenya, Alexander E; Rice, Charles M

    2004-11-19

    The NS5A protein of hepatitis C virus is believed to be an integral part of the viral replicase. Despite extensive investigation, the role of this protein remains elusive. Only limited biochemical characterization of NS5A has been performed, with most research to date involving the myriad of host proteins and signaling cascades that interact with NS5A. The need for better characterization of NS5A is paramount for elucidating the role of this protein in the virus life cycle. Examination of NS5A using bioinformatics tools suggested the protein consisted of three domains and contained an unconventional zinc binding motif within the N-terminal domain. We have developed a method to produce NS5A and performed limited proteolysis to confirm the domain organization model. The zinc content of purified NS5A and the N-terminal domain of NS5A was determined, and each of these proteins was found to coordinate one zinc atom per protein. The predicted zinc binding motif consists of four cysteine residues, conserved among the Hepacivirus and Pestivirus genera, fitting the formula of CX17CXCX20C. Mutation of any of the four cysteine components of this motif reduced NS5A zinc coordination and led to a lethal phenotype for HCV RNA replication, whereas mutation of other potential metal coordination residues in the N-terminal domain of NS5A, but outside the zinc binding motif, had little effect on zinc binding and, aside from one exception, were tolerated for replication. Collectively, these results indicate that NS5A is a zinc metalloprotein and that zinc coordination is likely required for NS5A function in the hepatitis C replicase.

  6. Daclatasvir inhibits hepatitis C virus NS5A motility and hyper-accumulation of phosphoinositides

    PubMed Central

    Chukkapalli, Vineela; Berger, Kristi L.; Kelly, Sean M.; Thomas, Meryl; Deiters, Alexander; Randall, Glenn

    2014-01-01

    Combinations of direct-acting antivirals (DAAs) against the hepatitis C virus (HCV) have the potential to revolutionize the HCV therapeutic regime. An integral component of DAA combination therapies are HCV NS5A inhibitors. It has previously been proposed that NS5A DAAs inhibit two functions of NS5A: RNA replication and virion assembly. In this study, we characterize the impact of a prototype NS5A DAA, daclatasvir (DCV), on HCV replication compartment formation. DCV impaired HCV replicase localization and NS5A motility. In order to characterize the mechanism behind altered HCV replicase localization, we examined the impact of DCV on the interaction of NS5A with its essential cellular cofactor, phosphatidylinositol-4-kinase III α (PI4KA). We observed that DCV does not inhibit PI4KA directly, nor does it impair early events of the NS5A-PI4KA interaction that can occur when NS5A is expressed alone. NS5A functions that are unaffected by DCV include PI4KA binding, as determined by co-immunoprecipitation, and a basal accumulation of the PI4KA product, PI4P. However, DCV impairs late steps in PI4KA activation that requires NS5A expressed in the context of the HCV polyprotein. These NS5A functions include hyper-stimulation of PI4P levels and appropriate replication compartment formation. The data are most consistent with a model wherein DCV inhibits conformational changes in the NS5A protein or protein complex formations that occur in the context of HCV polyprotein expression and stimulate PI4P hyper-accumulation and replication compartment formation. PMID:25546252

  7. Multifunctional adaptive NS1 mutations are selected upon human influenza virus evolution in the mouse.

    PubMed

    Forbes, Nicole E; Ping, Jihui; Dankar, Samar K; Jia, Jian-Jun; Selman, Mohammed; Keleta, Liya; Zhou, Yan; Brown, Earl G

    2012-01-01

    The role of the NS1 protein in modulating influenza A virulence and host range was assessed by adapting A/Hong Kong/1/1968 (H3N2) (HK-wt) to increased virulence in the mouse. Sequencing the NS genome segment of mouse-adapted variants revealed 11 mutations in the NS1 gene and 4 in the overlapping NEP gene. Using the HK-wt virus and reverse genetics to incorporate mutant NS gene segments, we demonstrated that all NS1 mutations were adaptive and enhanced virus replication (up to 100 fold) in mouse cells and/or lungs. All but one NS1 mutant was associated with increased virulence measured by survival and weight loss in the mouse. Ten of twelve NS1 mutants significantly enhanced IFN-β antagonism to reduce the level of IFN β production relative to HK-wt in infected mouse lungs at 1 day post infection, where 9 mutants induced viral yields in the lung that were equivalent to or significantly greater than HK-wt (up to 16 fold increase). Eight of 12 NS1 mutants had reduced or lost the ability to bind the 30 kDa cleavage and polyadenylation specificity factor (CPSF30) thus demonstrating a lack of correlation with reduced IFN β production. Mutant NS1 genes resulted in increased viral mRNA transcription (10 of 12 mutants), and protein production (6 of 12 mutants) in mouse cells. Increased transcription activity was demonstrated in the influenza mini-genome assay for 7 of 11 NS1 mutants. Although we have shown gain-of-function properties for all mutant NS genes, the contribution of the NEP mutations to phenotypic changes remains to be assessed. This study demonstrates that NS1 is a multifunctional virulence factor subject to adaptive evolution.

  8. H-NS Facilitates Sequence Diversification of Horizontally Transferred DNAs during Their Integration in Host Chromosomes

    PubMed Central

    Higashi, Koichi; Tobe, Toru; Kanai, Akinori; Uyar, Ebru; Ishikawa, Shu; Suzuki, Yutaka; Ogasawara, Naotake; Kurokawa, Ken; Oshima, Taku

    2016-01-01

    Bacteria can acquire new traits through horizontal gene transfer. Inappropriate expression of transferred genes, however, can disrupt the physiology of the host bacteria. To reduce this risk, Escherichia coli expresses the nucleoid-associated protein, H-NS, which preferentially binds to horizontally transferred genes to control their expression. Once expression is optimized, the horizontally transferred genes may actually contribute to E. coli survival in new habitats. Therefore, we investigated whether and how H-NS contributes to this optimization process. A comparison of H-NS binding profiles on common chromosomal segments of three E. coli strains belonging to different phylogenetic groups indicated that the positions of H-NS-bound regions have been conserved in E. coli strains. The sequences of the H-NS-bound regions appear to have diverged more so than H-NS-unbound regions only when H-NS-bound regions are located upstream or in coding regions of genes. Because these regions generally contain regulatory elements for gene expression, sequence divergence in these regions may be associated with alteration of gene expression. Indeed, nucleotide substitutions in H-NS-bound regions of the ybdO promoter and coding regions have diversified the potential for H-NS-independent negative regulation among E. coli strains. The ybdO expression in these strains was still negatively regulated by H-NS, which reduced the effect of H-NS-independent regulation under normal growth conditions. Hence, we propose that, during E. coli evolution, the conservation of H-NS binding sites resulted in the diversification of the regulation of horizontally transferred genes, which may have facilitated E. coli adaptation to new ecological niches. PMID:26789284

  9. A Hepatitis C Virus NS5A Phosphorylation Site That Regulates RNA Replication

    PubMed Central

    LeMay, K. L.; Treadaway, J.; Angulo, I.

    2013-01-01

    The hepatitis C virus NS5A protein is essential for RNA replication and virion assembly. NS5A is phosphorylated on multiple residues during infections, but these sites remain uncharacterized. Here we identify serine 222 of genotype 2a NS5A as a phosphorylation site that functions as a negative regulator of RNA replication. This site is a component of the hyperphosphorylated form of NS5A, which is in good agreement with previous observations that hyperphosphorylation negatively affects replication. PMID:23115292

  10. The GPM Common Calibrated Brightness Temperature Product

    NASA Astrophysics Data System (ADS)

    Stout, J.; Chou, J.

    2010-12-01

    The Global Precipitation Measurement (GPM) project will provide a core satellite carrying the GPM Microwave Imager (GMI) and will use microwave observations from a constellation of other satellites. Each partner with a satellite in the constellation will have a calibration that meets their own requirements and will decide on the format to archive their brightness temperature (Tb) record in GPM. However, GPM multi-sensor precipitation algorithms need to input intercalibrated Tb's in order to avoid differences among sensors introducing artifacts into the longer term climate record of precipitation. The GPM Common Calibrated Brightness Temperature Product is intended to address this problem by providing intercalibrated Tb data, called "Tc" data, where the "c" stands for common. The precipitation algorithms require a Tc file format that is both generic and flexible enough to accommodate the different passive microwave instruments. The format provides detailed information on the processing history in order to allow future researchers to have a record of what was done. The format is simple, including the main items of scan time, latitude, longitude, incidence angle, sun glint angle, and Tc. It also provides a quality flag, spacecraft orientation, spacecraft location, orbit, and instrument scan type (cross-track or conical). Another simplification is to store data in real numbers, avoiding the ambiguity of scaled data. Finally, units and descriptions will be provided in the product. The format is built on the concept of a swath, which is a series of scans that have common geolocation and common scan geometry. Scan geometry includes pixels per scan, sensor orientation, scan type, and incidence angles. The format includes 3 space saving methods: first rounding variables written as floats to their needed accuracy to achieve good compression, second writing sun glint angle as a one byte variable, and third storing only unique incidence angles but allowing access via a mapping

  11. Dark Skies, Bright Kids: Year 2

    NASA Astrophysics Data System (ADS)

    Carlberg, Joleen K.; Johnson, K.; Lynch, R.; Walker, L.; Beaton, R.; Corby, J.; de Messieres, G.; Drosback, M.; Gugliucci, N.; Jackson, L.; Kingery, A.; Layman, S.; Murphy, E.; Richardson, W.; Ries, P.; Romero, C.; Sivakoff, G.; Sokal, K.; Trammell, G.; Whelan, D.; Yang, A.; Zasowski, G.

    2011-01-01

    The Dark Skies, Bright Kids (DSBK) outreach program brings astronomy education into local elementary schools in central Virginia's Southern Albemarle County through an after-school club. Taking advantage of the unusually dark night skies in the rural countryside, DSBK targets economically disadvantaged schools that tend to be underserved due to their rural locale. The goals of DSBK are to foster children's natural curiosity, demonstrate that science is a fun and creative process, challenge students' conceptions of what a scientist is and does, and teach some basic astronomy. Furthermore, DSBK works to assimilate families into students' education by holding family observing nights at the school. Now in its third semester, DSBK has successfully run programs at two schools with very diverse student populations. Working with these students has helped us to revise our activities and to create new ones. A by-product of our work has been the development of lesson plans, complete with learning goals and detailed instructions, that we make publically available on our website. This year we are expanding our repertoire with our new planetarium, which allows us to visualize topics in novel ways and supplements family observing on cloudy nights. The DSBK volunteers have also created a bilingual astronomy artbook --- designed, written, and illustrated by UVa students --- that we will publish and distribute to elementary schools in Virginia. Our book debuted at the last AAS winter meeting, and since then it has been extensively revised and updated with input from many individuals, including parents, professional educators, and a children's book author. Because the club is currently limited to serving a few elementary schools, this book will be part of our efforts to broaden our impact by bringing astronomy to schools we cannot go to ourselves and reaching out to Spanish-speaking communities at the same time.

  12. Dark Skies, Bright Kids! Year 4

    NASA Astrophysics Data System (ADS)

    Sokal, Kimberly R.; Johnson, K. E.; Barcos-Munoz, L. D.; Beaton, R.; Borish, J.; Crawford, S. B.; Corby, J.; Damke, G.; Dean, J.; Dorsey, G.; Jackson, L.; Liss, S.; Oza, A.; Peacock, S.; Prager, B.; Romero, C.; Sivakoff, G. R.; Walker, L.; Whelan, D. G.; Zucker, C.

    2013-01-01

    Aiming to engage young children's natural excitement and curiosity, the outreach group Dark Skies, Bright Kids (DSBK) brings a hands-on approach to astronomy to elementary schools in Virginia. We hope to enhance children's view and understanding of science while exploring the Universe using fun activities. DSBK focuses on rural and underserved schools in Albemarle County and offers a semester-long astronomy club for third through fifth grade students. We believe regular interactions foster personal relationships between students and volunteers that encourage a life-long interest in science. In our fourth year of hosting clubs, we returned to Ivy Creek Elementary School, where we saw wonderful responses from a special group of students with `low-incidence' disabilities. DSBK has grown to realize a broader reach beyond local astronomy clubs; we hope to ignite a spark of interest in astronomy and science more widely- in more children, their families, and their teachers. We also hosted the Second Annual Central Virginia Star Party with an open invitation to the community to encourage families to enjoy astronomy together. Throughout the year, DSBK now holds 'one-off' programs (akin to astronomy field days) for elementary schools and children's groups throughout Virginia. Furthermore, we are in the final stages of a project to create two bilingual astronomy books called "Snapshots of the Universe", in Spanish and French with English translations. This art book will be made available online and we are working to get a copy in every elementary school in the state. DSBK has begun to reach out to elementary school teachers in order to provide them with useful and engaging classroom material. We have adapted our volunteer-created activities into useful and ready-to-use lessons, available online. After improvements based on research through interactions and feedback from teachers, we have explicitly identified the learning goals in terms of Virginia's Standards of Learning

  13. Dark Skies, Bright Kids! Year 5

    NASA Astrophysics Data System (ADS)

    Prager, Brian; Johnson, K. E.; Barcos-Munoz, L. D.; Beaton, R.; Bittle, L.; Borish, H.; Burkhardt, A.; Corby, J.; Damke, G.; Dean, J.; Dorsey, G.; Graninger, D.; Lauck, T.; Liss, S.; Oza, A.; Peacock, S.; Romero, C.; Sokal, K. R.; Stierwalt, S.; Walker, L.; Wenger, T.; Zucker, C.

    2014-01-01

    Our public outreach group Dark Skies, Bright Kids! (DSBK) fosters science literacy in Virginia by bringing a hands-on approach to astronomy that engages children's natural excitement and curiosity. We are an entirely volunteer-run group based out of the Department of Astronomy at the University of Virginia and we enthusiastically utilize astronomy as a 'gateway science.' We create long-term relationships with students during an 8 to 10 week long, after-school astronomy club at under served elementary schools in neighboring counties, and we visited 3 different schools in 2013. Additionally, we organize and participate in science events throughout the community. The fifth year of DSBK was marked by surpassing 10,000 contact hours in Spring 2013 Semester and by ringing in the fall semester with our biggest, most successful star party to date. We hosted the Third Annual Central Virginia Star Party, free and open to the community to encourage families to enjoy astronomy together. Nearly four hundred people of all ages attended, double the number from previous years. Joining with local astronomical societies, we offered an enlightening and exciting night with resources rarely accessible to the public, such as an IR camera and a portable planetarium. With numerous telescopes pointed at the sky, and a beautifully clear night with views of the Milky Way, the International Space Station, and numerous meteors, the star party was a fantastic opportunity to introduce many of our guests to the natural wonders of our night sky and enjoy some of the darkest skies on the eastern seaboard.

  14. Magnetic Flux Supplement to Coronal Bright Points

    NASA Astrophysics Data System (ADS)

    Mou, Chaozhou; Huang, Zhenghua; Xia, Lidong; Madjarska, Maria S.; Li, Bo; Fu, Hui; Jiao, Fangran; Hou, Zhenyong

    2016-02-01

    Coronal bright points (BPs) are associated with magnetic bipolar features (MBFs) and magnetic cancellation. Here we investigate how BP-associated MBFs form and how the consequent magnetic cancellation occurs. We analyze longitudinal magnetograms from the Helioseismic and Magnetic Imager to investigate the photospheric magnetic flux evolution of 70 BPs. From images taken in the 193 Å passband of the Atmospheric Imaging Assembly (AIA) we dermine that the BPs’ lifetimes vary from 2.7 to 58.8 hr. The formation of the BP MBFs is found to involve three processes, namely, emergence, convergence, and local coalescence of the magnetic fluxes. The formation of an MBF can involve more than one of these processes. Out of the 70 cases, flux emergence is the main process of an MBF buildup of 52 BPs, mainly convergence is seen in 28, and 14 cases are associated with local coalescence. For MBFs formed by bipolar emergence, the time difference between the flux emergence and the BP appearance in the AIA 193 Å passband varies from 0.1 to 3.2 hr with an average of 1.3 hr. While magnetic cancellation is found in all 70 BPs, it can occur in three different ways: (I) between an MBF and small weak magnetic features (in 33 BPs); (II) within an MBF with the two polarities moving toward each other from a large distance (34 BPs); (III) within an MBF whose two main polarities emerge in the same place simultaneously (3 BPs). While an MBF builds up the skeleton of a BP, we find that the magnetic activities responsible for the BP heating may involve small weak fields.

  15. Dark Skies, Bright Kids! Year 3

    NASA Astrophysics Data System (ADS)

    Whelan, David G.; Johnson, K. E.; Barcos-Munoz, L. D.; Beaton, R. L.; Borish, J.; Corby, J. F.; Dorsey, G.; Gugliucci, N. E.; Prager, B. J.; Ries, P. A.; Romero, C. E.; Sokal, K. R.; Tang, X.; Walker, L. M.; Yang, A. J.; Zasowski, G.

    2012-01-01

    Dark Skies, Bright Kids! (DSBK) is a program that brings astronomy education to elementary schools throughout central Virginia. In a relaxed, out-of-classroom atmosphere, we are able to foster the innate curiosity that young students have about science and the world around them. We target schools that are under-served due to their rural locale or special needs students, demonstrating that science is a fun and creative process to a segment of the population that might not otherwise be exposed to astronomy. Families are included in the learning experience during semi-annual `star parties'. Since last January, we have expanded the breadth and depth of our educational capabilities. We have developed new programs for use in our digital planetarium. We held the first Central Virginia Star Party, providing an atmosphere where local children from multiple schools were able to share their love for astronomy. Local government and University officials were also invited so that they could experience our focused science outreach. Most recently, we have become part of Ivy Creek School's Club Day activities, bringing our program to a new segment of the elementary school system in Albemarle County: those that have `low-incidence' disabilities, requiring special attention. We continue to develop a curriculum for after-school programs that functions as either a series of one-time activities or several months of focused outreach at one school. Many of these activities are provided on our website, http://www.astro.virginia.edu/dsbk/, for the wider astronomical community, including the new planetarium work. We have extended our book project to include two bilingual astronomy books called `Snapshots of the Universe,' one in Spanish and English, the other in French and English. These books introduce young people to some of the many wonders of the Universe through art and captions developed by DSBK volunteers.

  16. Easy way to estimate meteor brightness on TV frames

    NASA Astrophysics Data System (ADS)

    Leonov, V. A.; Bagrov, A. V.

    2016-01-01

    The traditional method of the meteor brightness measurements claims that the meteor brightness is equal to the stellar magnitude of a star that looks like a meteor in the brightest point of its track. This rule was convenient for the comparison of meteor observations by different observers and for the analysis of the brightness distributions of meteors from observed showers. This traditional method suffers from systematic errors, particularly those that arise from using stellar brightness measured in specific spectral wave bands different from the observer's ones, but mainly due to neglecting the influence of the meteor angular velocity on the real meteor brightness. To get a proper estimate of the meteor brightness that is a measure of the ground meteor illumination in the non-systematic units, an observer must take into account that the effective exposition of a meteor image in any resolution element of its track is a few times shorter than the corresponding exposition of a star image in the same frame. We propose a very simple method for improved estimations of meteor brightness by applying a correction to the meteor stellar magnitude obtained within the traditional framework.

  17. Bright Solitary Matter Waves: Formation, Stability and Interactions

    NASA Astrophysics Data System (ADS)

    Billam, T. P.; Marchant, A. L.; Cornish, S. L.; Gardiner, S. A.; Parker, N. G.

    In recent years, bright soliton-like structures composed of gaseous Bose-Einstein condensates have been generated at ultracold temperature. The experimental capacity to precisely engineer the nonlinearity and potential landscape experienced by these solitary waves offers an attractive platform for fundamental study of solitonic structures. The presence of three spatial dimensions and trapping implies that these are strictly distinct objects to the true soliton solutions. Working within the zero-temperature mean-field description, we explore the solutions and stability of bright solitary waves, as well as their interactions. Emphasis is placed on elucidating their similarities and differences to the true bright soliton. The rich behaviour introduced in the bright solitary waves includes the collapse instability and asymmetric collisions. We review the experimental formation and observation of bright solitary matter waves to date, and compare to theoretical predictions. Finally we discuss some topical aspects, including beyond-mean-field descriptions, symmetry breaking, exotic bright solitary waves, and proposals to exploit bright solitary waves in interferometry and as surface probes.

  18. Photometric Properties of Ceres and the Occator Bright Spots

    NASA Astrophysics Data System (ADS)

    Li, Jian-Yang; Le Corre, Lucille; Reddy, Vishnu; Sykes, Mark V.; Nathues, Andreas; Pieters, Carle M.; Ciarniello, Mauro; Turrini, Diego; McFadden, Lucy A.; Raymond, Carol A.; Russell, Christopher T.

    2015-11-01

    Dawn discovered several extremely bright spots on Ceres, the most prominent of which is located inside the Occator crater that is at least 4-5 times brighter than the average Ceres. Interestingly, these bright spots are located in relatively young craters that are at the longitudes corresponding to the maximum water vapor observed by the Herschel Space Observatory, suggesting possible correlation with water sublimation on Ceres. We used the multi-color imaging data collected by the Dawn Framing Camera to analyze the global photometric properties of Ceres and the bright spots, especially those located inside the Occator crater. Our objectives are to determine the albedo and other light scattering properties of the bright spots on Ceres in the visible wavelengths, in order to characterize their physical properties and find clues about their composition and possible formation mechanisms and the correlation with water sublimation. The overall geometric albedo of Ceres’ global surface is 0.09-0.10, consistent with previous studies. The Hapke roughness parameter is about 20°, close to many other asteroids, rather than 44° as reported earlier. Correspondingly, the phase function of Ceres is less backscattering than previously modeled. In contrast, the geometric albedo of the bright spots inside the Occator crater is 0.4-0.5, and the single scattering albedo is 0.7-0.8, brighter than Vesta’s global albedo but much darker than many icy satellites in the outer solar system. The Hapke roughness of the bright spots is much higher than Ceres average, suggesting relatively loose deposit of materials rather than more coherent or tightly packed materials. The phase function of bright spots material is relatively more forward scattering than average Ceres, possibly correlated to stronger multiple scattering due to high albedo resulting from more transparent materials. The highest resolution images as of late-August 2015 show fine structures within the Occator bright spots. We

  19. Development and clinical evaluation of a highly accurate dengue NS1 rapid test: from the preparation of a soluble NS1 antigen to the construction of an RDT.

    PubMed

    Lee, Jihoo; Kim, Hak-Yong; Chong, Chom-Kyu; Song, Hyun-Ok

    2015-06-01

    Early diagnosis of dengue virus (DENV) is important. There are numerous products on the market claiming to detect DENV NS1, but these are not always reliable. In this study, a highly sensitive and accurate rapid diagnostic test (RDT) was developed using anti-dengue NS1 monoclonal antibodies. A recombinant NS1 protein was produced with high antigenicity and purity. Monoclonal antibodies were raised against this purified NS1 antigen. The RDT was constructed using a capturing (4A6A10, Kd=7.512±0.419×10(-9)) and a conjugating antibody (3E12E6, Kd=7.032±0.322×10(-9)). The diagnostic performance was evaluated with NS1-positive clinical samples collected from various dengue endemic countries and compared to SD BioLine Dengue NS1 Ag kit. The constructed RDT exhibited higher sensitivity (92.9%) with more obvious diagnostic performance than the commercial kit (83.3%). The specificity of constructed RDT was 100%. The constructed RDT could offer a reliable point-of-care testing tool for the early detection of dengue infections in remote areas and contribute to the control of dengue-related diseases. PMID:25824725

  20. Development and clinical evaluation of a highly accurate dengue NS1 rapid test: from the preparation of a soluble NS1 antigen to the construction of an RDT.

    PubMed

    Lee, Jihoo; Kim, Hak-Yong; Chong, Chom-Kyu; Song, Hyun-Ok

    2015-06-01

    Early diagnosis of dengue virus (DENV) is important. There are numerous products on the market claiming to detect DENV NS1, but these are not always reliable. In this study, a highly sensitive and accurate rapid diagnostic test (RDT) was developed using anti-dengue NS1 monoclonal antibodies. A recombinant NS1 protein was produced with high antigenicity and purity. Monoclonal antibodies were raised against this purified NS1 antigen. The RDT was constructed using a capturing (4A6A10, Kd=7.512±0.419×10(-9)) and a conjugating antibody (3E12E6, Kd=7.032±0.322×10(-9)). The diagnostic performance was evaluated with NS1-positive clinical samples collected from various dengue endemic countries and compared to SD BioLine Dengue NS1 Ag kit. The constructed RDT exhibited higher sensitivity (92.9%) with more obvious diagnostic performance than the commercial kit (83.3%). The specificity of constructed RDT was 100%. The constructed RDT could offer a reliable point-of-care testing tool for the early detection of dengue infections in remote areas and contribute to the control of dengue-related diseases.

  1. Antiviral activities of 15 dengue NS2B-NS3 protease inhibitors using a human cell-based viral quantification assay.

    PubMed

    Chu, Justin Jang Hann; Lee, Regina Ching Hua; Ang, Melgious Jin Yan; Wang, Wei-Ling; Lim, Huichang Annie; Wee, John Liang Kuan; Joy, Joma; Hill, Jeffrey; Brian Chia, C S

    2015-06-01

    The dengue virus is a mosquito-borne pathogen responsible for an estimated 50-100 million human dengue infections annually. There are currently no approved drugs against this disease, resulting in a major unmet clinical need. The dengue viral NS2B-NS3 protease has been identified as a plausible drug target due to its involvement in viral replication in mammalian host cells. In the past decade, at least 20 dengue NS2B-NS3 protease inhibitors have been reported in the literature with a range of inhibitory activities in protease assays. However, such assays do not shed light on an inhibitor's ability to penetrate human cell membranes where the viral protease resides. In this study, we investigated the antiviral activities of 15 small-molecule and peptide-based NS2B-NS3 inhibitors on dengue serotype 2-infected HuH-7 human hepatocarcinoma cells. Experimental results revealed anthraquinone ARDP0006 (compound 5) to be the most potent inhibitor which reduced dengue viral titer by more than 1 log PFU/mL at 1 μM in our cell-based assays involving HuH-7 and K562 cell lines, suggesting that its scaffold could serve as a lead for further medicinal chemistry studies. Compound 5 was also found to be non-cytotoxic at 1 μM over 3 days incubation on HuH-7 cells using the Alamar Blue cellular toxicity assay.

  2. Degradation of Dye Wastewater by ns-Pulse DBD Plasma

    NASA Astrophysics Data System (ADS)

    Gao, Jin; Gu, Pingdao; Yuan, Li; Zhong, Fangchuan

    2013-09-01

    Two plasma reactors have been developed and used to degrade dye wastewater agents. The configuration of one plasma reactor is a comb-like extendable unit module consisting of 5 electrodes covered with a quartz tube and the other one is an array reactor which is extended from the unit module. The decomposition of wastewater by ns pulse dielectric barrier discharge (DBD) plasma have been carried out by atomizing the dyeing solutions into the reactors. During experiments, the indigo carmine has been treated as the waste agent. The measurements of UV-VIS absorption spectroscopy and the chemical oxygen demand (COD) are carried out to demonstrate the decomposition effect on the wastewater. It shows that the decoloration rate of 99% and the COD degradation rate of 65% are achieved with 15 min treatment in the unit reactor. The effect of electrical parameters on degradation has been studied in detail. Results from the array reactor indicate that it has a better degradation effect than the unit one. It can not only totally remove the chromogenic bond of the indigo carmine solution, but also effectively degrade unsaturated bonds. The decoloration rate reaches 99% after 10 min treatment, the decomposition rate of the unsaturated bond reaches 83% after 60 min treatment, and the COD degradation rate is nearly 74%.

  3. Development of parallel incompressible NS solver on stretched grids

    NASA Astrophysics Data System (ADS)

    Jothiprasad, G.; Caughey, D.; Pope, S. B.

    2003-11-01

    Development of a parallel NS solver for studying DNS and LES of temporal mixing layers is discussed. The equations are cast in strong conservation form on a uniform computational mesh, transformed from a stretched mesh in the physical domain. Variables are defined on a collocated grid, and the transformed equations are solved using a fractional step method. Convective and dissipative terms are treated using explicit Adams-Bashforth and implicit Crank-Nicolson, respectively. Fourth order spatial accuracy is maintained except for hyperviscous subgrid model terms, which are only 2nd order accurate. The block LU analysis of J. B. Perot, extended to fractional step methods on collocated grids, shows that an O(Δ t^2) term involving the pressure gradient must be added to the momentum equations to maintain 2nd order accuracy in time. Using a smaller stencil for the pressure gradients largely simplifies the pressure Poisson equation while still ensuring that discrete continuity is satisfied to appropriate order. Implementation on distributed-memory multiprocessors is achieved using MPI, with care taken to minimize communication overhead.

  4. Comments on D-brane dynamics near NS5-branes

    NASA Astrophysics Data System (ADS)

    Sahakyan, David A.

    2004-10-01

    We study the properties of a D-brane in the presence of k NS5 branes. The Dirac-Born-Infeld action describing the dynamics of this D-brane is very similar to that of a non-BPS D-brane in ten dimensions. As the D-brane approaches the fivebranes, its equation of state approaches that of a pressureless fluid. In non-BPS D-brane case this is considered as an evidence for the decay of the D-brane into ``tachyon matter''. We show that in our case similar behavior is the consequence of the motion of the D-brane. In particular in the rest frame of the moving D-brane the equation of state is that of a usual D-brane, for which the pressure is equal to the energy density. We also compute the total cross-section for the decay of the D-brane into closed string modes and show that the emitted energy has a power like divergence for D0, D1 and D2 branes, while converges for higher dimensional D-branes. We also speculate on the possibility that the infalling D-brane describes a decaying defect in six dimensional Little String Theory.

  5. Nitrogen oxide removal dynamic process through 15 Ns DBD technique

    NASA Astrophysics Data System (ADS)

    Wang, Xiaojun; Zhang, Lianshui; Lai, Weidong; Liu, Fengliang

    2015-05-01

    Nitrogen oxides exhaust gas assumes the important responsibility on air pollution by forming acid rain. This paper discusses the NO removal mechanism in 15 ns pulse dielectric barrier discharge (DBD) plasma through experimental and simulating method. Emission spectra collected from plasma are evaluated as sourced from N+ and O(3P). The corresponding zero-dimensional model is established and verified through comparing the simulated concentration evolution and the experimental time-resolved spectra of N+. The electron impact ionization plays major role on NO removal and the produced NO+ are further decomposed into N+ and O(3P) through electron impact dissociative excitation rather than the usual reported dissociative recombination process. Simulation also indicates that the removal process can be accelerated by NO inputted at lower initial concentration or electrons streamed at higher concentration, due to the heightened electron impact probability on NO molecules. The repetitive pulse discharge is a benefit for improving the NO removal efficiency by effectively utilizing the radicals generated from the previous pulse under the condition that the pulse period should be shorter enough to ignore the spatial diffusion of radicals. Finally, slight attenuation on NO removal has been experimentally and simulatively observed after N2 mixed, due to the competitive consumption of electrons.

  6. A new panel of NS1 antibodies for easy detection and titration of influenza A virus.

    PubMed

    Tan, Zhihao; Akerstrom, Sara; Wee, Boon Yu; Lal, Sunil K; Mirazimi, Ali; Tan, Yee-Joo

    2010-03-01

    The non-structural protein NS1 of the influenza A virus is a good target for the development of diagnostic assays. In this study, three NS1 monoclonal antibodies (mAbs) were generated by using recombinant NS1 protein of H5N1 virus and found to bind both the native and denatured forms of NS1. Two of the mAbs, 6A4 and 2H6, bind NS1 of three different strains of influenza A virus, namely H1N1, H3N2, and H5N1. Epitope mapping revealed that residues 42-53 of H5N1 NS1 are essential for the interaction with both mAbs. Between the three strains, there is only one amino acid difference in this domain, which is consistent with the observed cross-reactivities. On the other hand, mAb 1G1 binds to residues 206-215 of H5N1 NS1 and does not bind NS1 of H1N1 or H3N2. Furthermore, all three mAbs detected NS1 proteins expressed in virus infected MDCK cells and indirect immunofluorescence staining with mAbs 6A4 and 2H6 provided an alternative method for viral titer determination. Quantifying the numbers of fluorescent foci units yielded viral titers for three different isolates of H5N1 virus that are highly comparable to that obtained by observing cytopathic effect induced by virus infection. Importantly, this alternative method yields results at 1 day post-infection while the conventional method using cytopathic effect yields results at 3 days post-infection. The results showed that this new panel of NS1 antibodies can detect NS1 protein expressed during viral infection and can be used for fast and easy titration of influenza A virus. J. Med. Virol. 82:467-475, 2010. (c) 2010 Wiley-Liss, Inc.

  7. Brightness field distributions of microlens arrays using micro molding.

    PubMed

    Cheng, Hsin-Chung; Huang, Chiung-Fang; Lin, Yi; Shen, Yung-Kang

    2010-12-20

    This study describes the brightness field distributions of microlens arrays fabricated by micro injection molding (μIM) and micro injection-compression molding (μICM). The process for fabricating microlens arrays used room-temperature imprint lithography, photoresist reflow, electroforming, μIM, μICM, and optical properties measurement. Analytical results indicate that the brightness field distribution of the molded microlens arrays generated by μICM is better than those made using μIM. Our results further demonstrate that mold temperature is the most important processing parameter for brightness field distribution of molded microlens arrays made by μIM or μICM.

  8. HUBBLE FINDS MANY BRIGHT CLOUDS ON URANUS

    NASA Technical Reports Server (NTRS)

    2002-01-01

    A recent Hubble Space Telescope view reveals Uranus surrounded by its four major rings and by 10 of its 17 known satellites. This false-color image was generated by Erich Karkoschka using data taken on August 8, 1998, with Hubble's Near Infrared Camera and Multi-Object Spectrometer. Hubble recently found about 20 clouds - nearly as many clouds on Uranus as the previous total in the history of modern observations. The orange-colored clouds near the prominent bright band circle the planet at more than 300 mph (500 km/h), according to team member Heidi Hammel (MIT). One of the clouds on the right-hand side is brighter than any other cloud ever seen on Uranus. The colors in the image indicate altitude. Team member Mark Marley (New Mexico State University) reports that green and blue regions show where the atmosphere is clear and sunlight can penetrate deep into Uranus. In yellow and grey regions the sunlight reflects from a higher haze or cloud layer. Orange and red colors indicate very high clouds, such as cirrus clouds on Earth. The Hubble image is one of the first images revealing the precession of the brightest ring with respect to a previous image [LINK to PRC97-36a]. Precession makes the fainter part of the ring (currently on the upper right-hand side) slide around Uranus once every nine months. The fading is caused by ring particles crowding and hiding each other on one side of their eight-hour orbit around Uranus. The blue, green and red components of this false-color image correspond to exposures taken at near-infrared wavelengths of 0.9, 1.1, and 1.7 micrometers. Thus, regions on Uranus appearing blue, for example, reflect more sunlight at 0.9 micrometer than at the longer wavelengths. Apparent colors on Uranus are caused by absorption of methane gas in its atmosphere, an effect comparable to absorption in our atmosphere which can make distant clouds appear red. Credit: Erich Karkoschka (University of Arizona) and NASA

  9. Experimental evidence and molecular modeling of the interaction between hRSV-NS1 and quercetin.

    PubMed

    Gomes, Deriane Elias; Caruso, Ícaro Putinhon; de Araujo, Gabriela Campos; de Lourenço, Isabella Otenio; de Melo, Fernando Alves; Cornélio, Marinônio Lopes; Fossey, Marcelo Andrés; de Souza, Fátima Pereira

    2016-04-01

    Human Respiratory Syncytial Virus is one of the major causes of acute respiratory infections in children, causing bronchiolitis and pneumonia. Non-Structural Protein 1 (NS1) is involved in immune system evasion, a process that contributes to the success of hRSV replication. This protein can act by inhibiting or neutralizing several steps of interferon pathway, as well as by silencing the hRSV ribonucleoproteic complex. There is evidence that quercetin can reduce the infection and/or replication of several viruses, including RSV. The aims of this study include the expression and purification of the NS1 protein besides experimental and computational assays of the NS1-quercetin interaction. CD analysis showed that NS1 secondary structure composition is 30% alpha-helix, 21% beta-sheet, 23% turn and 26% random coils. The melting temperature obtained through DSC analysis was around 56°C. FRET analysis showed a distance of approximately 19Å between the NS1 and quercetin. Fluorescence titration results showed that the dissociation constant of the NS1-quercetin interaction was around 10(-6)M. In thermodynamic analysis, the enthalpy and entropy balanced forces indicated that the NS1-quercetin interaction presented both hydrophobic and electrostatic contributions. The computational results from the molecular modeling for NS1 structure and molecular docking regarding its interaction with quercetin corroborate the experimental data.

  10. Identification and characterization of coumestans as novel HCV NS5B polymerase inhibitors

    PubMed Central

    Kaushik-Basu, Neerja; Bopda-Waffo, Alain; Talele, Tanaji T.; Basu, Amartya; Costa, Paulo R. R.; da Silva, Alcides J. M.; Sarafianos, Stefan G.; Noël, François

    2008-01-01

    The hepatitis C virus (HCV) NS5B is essential for viral RNA replication and is therefore a prime target for development of HCV replication inhibitors. Here, we report the identification of a new class of HCV NS5B inhibitors belonging to the coumestan family of phytoestrogens. Based on the in vitro NS5B RNA-dependent RNA polymerase (RdRp) inhibition in the low micromolar range by wedelolactone, a naturally occurring coumestan, we evaluated the anti-NS5B activity of four synthetic coumestan analogues bearing different patterns of substitutions in their A and D rings, and observed a good structure-activity correlation. Kinetic characterization of coumestans revealed a noncompetitive mode of inhibition with respect to nucleoside triphosphate (rNTP) substrate and a mixed mode of inhibition towards the nucleic acid template, with a major competitive component. The modified order of addition experiments with coumestans and nucleic acid substrates affected the potencies of the coumestan inhibitors. Coumestan interference at the step of NS5B–RNA binary complex formation was confirmed by cross-linking experiments. Molecular docking of coumestans within the allosteric site of NS5B yielded significant correlation between their calculated binding energies and IC50 values. Coumestans thus add to the diversifying pool of anti-NS5B agents and provide a novel scaffold for structural refinement and development of potent NS5B inhibitors. PMID:18203743

  11. Canine parvovirus NS1 protein exhibits anti-tumor activity in a mouse mammary tumor model.

    PubMed

    Gupta, Shishir Kumar; Yadav, Pavan Kumar; Gandham, Ravi Kumar; Sahoo, A P; Harish, D R; Singh, Arvind Kumar; Tiwari, A K

    2016-02-01

    Many viral proteins have the ability to kill tumor cells specifically without harming the normal cells. These proteins, on ectopic expression, cause lysis or induction of apoptosis in the target tumor cells. Parvovirus NS1 is one of such proteins, which is known to kill high proliferating tumor cells. In the present study, we assessed the apoptosis inducing ability of canine parvovirus type 2 NS1 protein (CPV2.NS1) in vitro in 4T1 cells, and found it to cause significant cell death due to induction of apoptosis through intrinsic or mitochondrial pathway. Further, we also evaluated the oncolytic activity of CPV2.NS1 protein in a mouse mammary tumor model. The results suggested that CPV2.NS1 was able to inhibit the growth of 4T1 induced mouse mammary tumor as indicated by significantly reduced tumor volume, mitotic, AgNOR and PCNA indices. Further, inhibition of tumor growth was found to be because of induction of apoptosis in the tumor cells, which was evident by a significant increase in the number of TUNEL positive cells. Further, CPV2.NS1 was also able to stimulate the immune cells against the tumor antigens as indicated by the increased CD4+ and CD8+ counts in the blood of CVP2.NS1 treated mice. Further optimization of the delivery of NS1 protein and use of an adjuvant may further enhance its anti-tumor activity.

  12. NS1-mediated delay of type I interferon induction contributes to influenza A virulence in ferrets.

    PubMed

    Meunier, Isabelle; von Messling, Veronika

    2011-07-01

    Interference of the influenza A virus non-structural protein NS1 with type I interferon (IFN) signalling has been characterized extensively in vitro. To assess the contribution of NS1 to the virulence of a specific strain, we generated recombinant USSR/90/77 viruses bearing the NS1 proteins of the attenuated strain PR/8/34 or the highly pathogenic strain 1918 'Spanish flu', all belonging to the H1N1 subtype. In vitro, the extent of interference with type I IFN production exerted by the different NS1 proteins correlated with the reported virulence of the respective strain. Infection of ferrets with the recombinant viruses revealed that the presence of the 1918 NS1 resulted in a slightly more severe disease with generally higher clinical scores and increased lung pathology. Analysis of mRNA from nasal wash cells revealed that viruses carrying the 1918 and, to a lesser extent, USSR/90/77 NS1 proteins caused a delay in upregulation of type I IFNs compared with the NS1 PR/8/34-expressing virus, demonstrating the importance of NS1 for early host-response control and virulence.

  13. Influenza virus non-structural protein NS1: interferon antagonism and beyond.

    PubMed

    Marc, Daniel

    2014-12-01

    Most viruses express one or several proteins that counter the antiviral defences of the host cell. This is the task of non-structural protein NS1 in influenza viruses. Absent in the viral particle, but highly expressed in the infected cell, NS1 dramatically inhibits cellular gene expression and prevents the activation of key players in the IFN system. In addition, NS1 selectively enhances the translation of viral mRNAs and may regulate the synthesis of viral RNAs. Our knowledge of the virus and of NS1 has increased dramatically during the last 15 years. The atomic structure of NS1 has been determined, many cellular partners have been identified and its multiple activities have been studied in depth. This review presents our current knowledge, and attempts to establish relationships between the RNA sequence, the structure of the protein, its ligands, its activities and the pathogenicity of the virus. A better understanding of NS1 could help in elaborating novel antiviral strategies, based on either live vaccines with altered NS1 or on small-compound inhibitors of NS1.

  14. A Second RNA-Binding Site in the NS1 Protein of Influenza B Virus.

    PubMed

    Ma, Li-Chung; Guan, Rongjin; Hamilton, Keith; Aramini, James M; Mao, Lei; Wang, Shanshan; Krug, Robert M; Montelione, Gaetano T

    2016-09-01

    Influenza viruses cause a highly contagious respiratory disease in humans. The NS1 proteins of influenza A and B viruses (NS1A and NS1B proteins, respectively) are composed of two domains, a dimeric N-terminal domain and a C-terminal domain, connected by a flexible polypeptide linker. Here we report the 2.0-Å X-ray crystal structure and nuclear magnetic resonance studies of the NS1B C-terminal domain, which reveal a novel and unexpected basic RNA-binding site that is not present in the NS1A protein. We demonstrate that single-site alanine replacements of basic residues in this site lead to reduced RNA-binding activity, and that recombinant influenza B viruses expressing these mutant NS1B proteins are severely attenuated in replication. This novel RNA-binding site of NS1B is required for optimal influenza B virus replication. Most importantly, this study reveals an unexpected RNA-binding function in the C-terminal domain of NS1B, a novel function that distinguishes influenza B viruses from influenza A viruses.

  15. A Second RNA-Binding Site in the NS1 Protein of Influenza B Virus.

    PubMed

    Ma, Li-Chung; Guan, Rongjin; Hamilton, Keith; Aramini, James M; Mao, Lei; Wang, Shanshan; Krug, Robert M; Montelione, Gaetano T

    2016-09-01

    Influenza viruses cause a highly contagious respiratory disease in humans. The NS1 proteins of influenza A and B viruses (NS1A and NS1B proteins, respectively) are composed of two domains, a dimeric N-terminal domain and a C-terminal domain, connected by a flexible polypeptide linker. Here we report the 2.0-Å X-ray crystal structure and nuclear magnetic resonance studies of the NS1B C-terminal domain, which reveal a novel and unexpected basic RNA-binding site that is not present in the NS1A protein. We demonstrate that single-site alanine replacements of basic residues in this site lead to reduced RNA-binding activity, and that recombinant influenza B viruses expressing these mutant NS1B proteins are severely attenuated in replication. This novel RNA-binding site of NS1B is required for optimal influenza B virus replication. Most importantly, this study reveals an unexpected RNA-binding function in the C-terminal domain of NS1B, a novel function that distinguishes influenza B viruses from influenza A viruses. PMID:27545620

  16. Reverting cholesterol auxotrophy of NS0 cells by altering epigenetic gene silencing.

    PubMed

    Seth, Gargi; Ozturk, Mustafa; Hu, Wei-Shou

    2006-03-01

    NS0 is a cholesterol-requiring mouse myeloma cell line widely used in the production of recombinant antibodies. We have previously reported that the deficiency of 17beta-hydroxysteroid dehydrogenase type7 (Hsd17b7) is responsible for the cholesterol auxotrophy of NS0 cells. Here we demonstrate DNA methylation to be the mechanism underlying transcriptional suppression of Hsd17b7 in cholesterol dependent NS0 cells. Analysis of the DNA methylation pattern revealed methylation of the CpG-rich region upstream of the Hsd17b7 transcription start site in NS0 cells. This is in contrast to the unmethylated status of this sequence in a naturally isolated cholesterol independent revertant cell population (NS0_r). This transcriptional repression was relieved after treating cells with the demethylating drug, 5-azacytidine. Drug treatment also gave rise to high frequency cholesterol-independent variants. Characterization of revertants revealed substantially elevated transcript level of 17beta-hydroxysteroid dehydrogenase type7 (Hsd17b7) gene along with hypomethylation of the CpG-rich region. These results affirm that deficiency of Hsd17b7 causes cholesterol dependence of NS0 cells. Furthermore, induction of cholesterol independence by altering DNA methylation pattern alludes to the role of epigenetics in the metabolic adaptation of NS0 cells. With the widespread use of NS0 cells, this finding will have a significant impact on the optimization of recombinant antibody production processes. PMID:16189819

  17. Identification of an NTPase motif in classical swine fever virus NS4B protein

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Classical swine fever (CSF) is a highly contagious and often fatal disease of swine caused by CSF virus (CSFV), a positive sense single-stranded RNA virus in the genus Pestivirus of the Flaviviridae family. Here, we have identified, within CSFV non-structural (NS) protein NS4B, conserved sequence el...

  18. Mutations in the classical swine fever virus NS4B protein affects virulence in swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    NS4B is one of the non-structural proteins of Classical Swine Fever Virus (CSFV), the etiological agent of a severe, highly lethal disease of swine. Protein domain analysis of the predicted amino acid sequence of the NS4B protein of highly pathogenic CSFV strain Brescia (BICv) identified a Toll/Inte...

  19. Visible Color and Photometry of Bright Materials on Vesta

    NASA Technical Reports Server (NTRS)

    Schroder, S. E.; Li, J. Y.; Mittlefehldt, D. W.; Pieters, C. M.; De Sanctis, M. C.; Hiesinger, H.; Blewett, D. T.; Russell, C. T.; Raymond, C. A.; Keller, H. U.

    2012-01-01

    The Dawn Framing Camera (FC) collected images of the surface of Vesta at a pixel scale of 70 m in the High Altitude Mapping Orbit (HAMO) phase through its clear and seven color filters spanning from 430 nm to 980 nm. The surface of Vesta displays a large diversity in its brightness and colors, evidently related to the diverse geology [1] and mineralogy [2]. Here we report a detailed investigation of the visible colors and photometric properties of the apparently bright materials on Vesta in order to study their origin. The global distribution and the spectroscopy of bright materials are discussed in companion papers [3, 4], and the synthesis results about the origin of Vestan bright materials are reported in [5].

  20. Detail of Bright Angel stone vault, containing condenser, Hoffman condensation ...

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

    Detail of Bright Angel stone vault, containing condenser, Hoffman condensation pump, Jennings vacuum heating pump, and misc. pipes and valves. - Grand Canyon Village Utilities, Grand Canyon National Park, Grand Canyon Village, Coconino County, AZ

  1. IR brightness and eclipse cooling of Saturn's rings

    NASA Technical Reports Server (NTRS)

    Nolt, I. G.; Caldwell, J.; Radostitz, J. V.; Tokunaga, A. T.; Barrett, E. W.; Gillett, F. C.; Murphy, R. E.

    1980-01-01

    Equatorial scans of Saturn at 20 microns wavelength, obtained with the Mayall 4-m telescope in 1978-79, show a continuing decrease in the specific brightness of the A and B rings as the ring plane projection approaches and edge-on apparition. The decreased brightness of the previously dominant B ring reveals more clearly a large difference in brightness between the east and west ansae portions of the C ring, in contrast to a barely discernible difference for the other ring ansae. The amount of eclipse cooling is compatible with a C ring particle size of about 1 cm. It is proposed here that the B ring brightness variation could partially result from a decrease of absorbed insolation by a modest amount of visible scattering.

  2. Direct LED backlight for large area LCD TVs: brightness analysis

    NASA Astrophysics Data System (ADS)

    Sun, Ching-Cherng; Moreno, Ivan; Chung, Shih-Hsun; Chien, Wei-Ting; Hsieh, Chih-To; Yang, Tsung-Hsun

    2007-09-01

    A direct or bottom LED backlight is a key concept in large area LCD displays because it does not use a light guide, is flat, and is easy to assemble. In this paper, a method of luminance management for a bottom LED backlight is proposed and demonstrated. We analytically calculate both the power consumption and brightness uniformity in function of: screen brightness, screen size, backlight thickness, transmittance of the LCD panel, reflective cavity efficiency, gain and cone angle of enhancement films, LED array configuration, and the average luminous flux and radiation pattern of a single LED. Moreover, a 42-inch LCD television with this backlight design approach is made and demonstrated. The bottom backlight incorporates an array of RGGB 4-in-1 multi-chip LEDs within a highly reflective box behind a diffuser and a dual brightness enhancement film. We predict with an accuracy of 94% the brightness uniformity and with 96% the luminance level.

  3. Yellow steam and electrical pipes across from Bright Angel Lodge. ...

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

    Yellow steam and electrical pipes across from Bright Angel Lodge. Note control valve to right of control box, view E. - Grand Canyon Village Utilities, Grand Canyon National Park, Grand Canyon Village, Coconino County, AZ

  4. Perspective of Bright Angel stone vault, view south, with HAER ...

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

    Perspective of Bright Angel stone vault, view south, with HAER field team measuring (Michael Lee and Dominic Duran foreground, Christopher Marston rear). - Grand Canyon Village Utilities, Grand Canyon National Park, Grand Canyon Village, Coconino County, AZ

  5. Bright Angel stone vault, with HAER field team members Dominic ...

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

    Bright Angel stone vault, with HAER field team members Dominic Duran, Christopher Marston, and Michael Lee (l to r). - Grand Canyon Village Utilities, Grand Canyon National Park, Grand Canyon Village, Coconino County, AZ

  6. South and west elevations of Bright Angel boiler house. Red ...

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

    South and west elevations of Bright Angel boiler house. Red Horse log cabin visible in background. - Grand Canyon Village Utilities, Grand Canyon National Park, Grand Canyon Village, Coconino County, AZ

  7. Phosphorylation of influenza A virus NS1 protein at threonine 49 suppresses its interferon antagonistic activity

    PubMed Central

    Kathum, Omer Abid; Schräder, Tobias; Anhlan, Darisuren; Nordhoff, Carolin; Liedmann, Swantje; Pande, Amit; Mellmann, Alexander; Ehrhardt, Christina; Wixler, Viktor

    2016-01-01

    Summary Phosphorylation and dephosphorylation acts as a fundamental molecular switch that alters protein function and thereby regulates many cellular processes. The non‐structural protein 1 (NS1) of influenza A virus is an important factor regulating virulence by counteracting cellular immune responses against viral infection. NS1 was shown to be phosphorylated at several sites; however, so far, no function has been conclusively assigned to these post‐translational events yet. Here, we show that the newly identified phospho‐site threonine 49 of NS1 is differentially phosphorylated in the viral replication cycle. Phosphorylation impairs binding of NS1 to double‐stranded RNA and TRIM25 as well as complex formation with RIG‐I, thereby switching off its interferon antagonistic activity. Because phosphorylation was shown to occur at later stages of infection, we hypothesize that at this stage other functions of the multifunctional NS1 beyond its interferon‐antagonistic activity are needed. PMID:26687707

  8. Monoclonal antibodies against NS4B protein of japanese encephalitis virus.

    PubMed

    Ruan, Xindi; Huang, Shaomei; Shao, Lin; Ye, Jing; Chen, Zheng; Chen, Huanchun; Cao, Shengbo

    2013-12-01

    Japanese encephalitis (JE) is one of the most prevalent global viral encephalitis viruses. The functions of JEV (virus) NS4B protein are still under investigation. In our study, NS4B was expressed in Escherichia coli and purified by dialysis. Two clones of monoclonal antibodies (MAbs 1B1 and 1C3) against NS4B protein were generated and their characterizations were investigated. IFA, Western blot, and ELISA results showed that the MAbs were specific against JEV NS4B protein. The epitope of the MAbs was further identified using pairs of synthesized overlapping peptides. These MAbs may provide valuable tools for further exploration of the functions of NS4B and the pathogenesis of Japanese encephalitis virus. PMID:24328740

  9. Phosphorylation of influenza A virus NS1 protein at threonine 49 suppresses its interferon antagonistic activity.

    PubMed

    Kathum, Omer Abid; Schräder, Tobias; Anhlan, Darisuren; Nordhoff, Carolin; Liedmann, Swantje; Pande, Amit; Mellmann, Alexander; Ehrhardt, Christina; Wixler, Viktor; Ludwig, Stephan

    2016-06-01

    Phosphorylation and dephosphorylation acts as a fundamental molecular switch that alters protein function and thereby regulates many cellular processes. The non-structural protein 1 (NS1) of influenza A virus is an important factor regulating virulence by counteracting cellular immune responses against viral infection. NS1 was shown to be phosphorylated at several sites; however, so far, no function has been conclusively assigned to these post-translational events yet. Here, we show that the newly identified phospho-site threonine 49 of NS1 is differentially phosphorylated in the viral replication cycle. Phosphorylation impairs binding of NS1 to double-stranded RNA and TRIM25 as well as complex formation with RIG-I, thereby switching off its interferon antagonistic activity. Because phosphorylation was shown to occur at later stages of infection, we hypothesize that at this stage other functions of the multifunctional NS1 beyond its interferon-antagonistic activity are needed.

  10. Structure of NS1A effector domain from the influenza A/Udorn/72 virus.

    PubMed

    Xia, Shuangluo; Monzingo, Arthur F; Robertus, Jon D

    2009-01-01

    The nonstructural protein NS1A from influenza virus is a multifunctional virulence factor and a potent inhibitor of host immunity. It has two functional domains: an N-terminal 73-amino-acid RNA-binding domain and a C-terminal effector domain. Here, the crystallographic structure of the NS1A effector domain of influenza A/Udorn/72 virus is presented. Structure comparison with the NS1 effector domain from mouse-adapted influenza A/Puerto Rico/8/34 (PR8) virus strain reveals a similar monomer conformation but a different dimer interface. Further analysis and evaluation shows that the dimer interface observed in the structure of the PR8 NS1 effector domain is likely to be a crystallographic packing effect. A hypothetical model of the intact NS1 dimer is presented.

  11. BOREAS Level-2 NS001 TMS Imagery: Reflectance and Temperature in BSQ Format

    NASA Technical Reports Server (NTRS)

    Lobitz, Brad; Spanner, Michael; Hall, Forrest G. (Editor); Newcomer, Jeffrey A. (Editor); Strub, Richard

    2000-01-01

    For BOREAS, the NS001 TMS images, along with the other remotely sensed data, were collected to provide spatially extensive information over the primary study areas. This information includes detailed land cover and biophysical parameter maps such as fPAR and LAI. Collection of the NS001 images occurred over the study areas during the 1994 field campaigns. The level-2 NS001 data are atmospherically corrected versions of some of the best original NS001 imagery and cover the dates of 19-Apr-1994, 07-Jun-1994, 21-Jul-1994, 08-Aug-1994, and 16-Sep-1994. The data are not geographically/geometrically corrected; however, files of relative X and Y coordinates for each image pixel were derived by using the C130 INS data in an NS001 scan model. The data are provided in binary image format files.

  12. Structure and Function of Flavivirus NS5 Methyltransferase

    SciTech Connect

    Zhou,Y.; Ray, D.; Zhao, Y.; Dong, H.; Ren, S.; Li, Z.; Guo, Y.; Bernard, K.; Shi, P.; Li, H.

    2007-01-01

    The plus-strand RNA genome of flavivirus contains a 5' terminal cap 1 structure (m{sup 7}GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2'-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA {yields} m{sup 7}GpppA {yields} m{sup 7}GpppAm. The 2'-O methylation can be uncoupled from the N-7 methylation, since m{sup 7}GpppA-RNA can be readily methylated to m{sup 7}GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 {angstrom} resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2'-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltransferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2'-O cap methylations require distinct buffer conditions and different side chains within the K{sub 61}-D{sub 146}-K{sub 182}-E{sub 218} motif, suggesting that the two reactions use different mechanisms. In the context of complete virus, defects in both methylations are lethal to WNV; however, viruses defective solely in 2'-O methylation are attenuated and can protect mice from later wild-type WNV challenge. The results demonstrate that the N-7 methylation activity is essential for the WNV life cycle and, thus, methyltransferase represents a novel target for flavivirus therapy.

  13. Hepatitis C Virus NS3 Mutations in Hemophiliacs

    PubMed Central

    Lin, Ming Valerie; Charlton, Ashley N.; Rouster, Susan D.; Zamor, Philippe J.; Sherman, Kenneth E.

    2014-01-01

    Introduction Hemophiliacs have high HCV exposure risk from blood products that did not undergo heat inactivation or disease-specific screening prior to 1987. Repeated exposure to infected factor concentrates predisposes hemophiliacs to higher likelihood of HCV from multiple sources. HIV coinfection could result in impaired clearance of less fit variants resulting in enrichment of quasispecies carrying resistance mutations. Aim We postulated that hemophiliacs demonstrate increased prevalence of baseline signature mutations in the HCV NS3/4 serine protease coding domain. Methods We examined the prevalence of putative HCV protease inhibitor mutations, mutations, sub-classified into dominant mutations if changes conferred resistance, and minor variants not associated with drug resistance, in patients with hemophilia A or B, infected with HCV or HCV/HIV, prior to HCV PI exposure. Results 151 subjects were evaluated, including 22 hemophiliacs and 129 non-hemophilic controls. Of 58 mutations detected, 55 (95%) were resistance mutations and 3 (5%) were minor variants. Dominant mutations were detected in 10 (45.5%) hemophiliacs and in 43 (33.3%) controls (OR 1.67, 95% CI 0.67–4.16). There was no statistical difference in proportion of dominant mutations (p=0.27) or minor variants (p=0.47) between groups, despite adjustment for HIV status (p=0.44). Conclusion No significant differences in dominant or minor resistance mutations between hemophiliacs and non-hemophiliacs were observed. HIV presence or prior HAART exposure did not affect baseline distribution. We conclude that hemophiliacs are not at higher risk for pre-existing HCV PI mutations, and prospective studies of response to PI-based regimens with HCV activity are indicated. PMID:24697920

  14. Species specificity of the NS1 protein of influenza B virus: NS1 binds only human and non-human primate ubiquitin-like ISG15 proteins.

    PubMed

    Sridharan, Haripriya; Zhao, Chen; Krug, Robert M

    2010-03-12

    Influenza B viruses, which cause a highly contagious respiratory disease every year, are restricted to humans, but the basis for this restriction had not been determined. Here we provide one explanation for this restriction: the species specificity exhibited by the NS1 protein of influenza B virus (NS1B protein). This viral protein combats a major host antiviral response by binding the interferon-alpha/beta-induced, ubiquitin-like ISG15 protein and inhibiting its conjugation to an array of proteins. We demonstrate that the NS1B protein exhibits species-specific binding; it binds human and non-human primate ISG15 but not mouse or canine ISG15. In both transfection assays and virus-infected cells, the NS1B protein binds and relocalizes only human and non-human primate ISG15 from the cytoplasm to nuclear speckles. Human and non-human primate ISG15 proteins consist of two ubiquitin-like domains separated by a short hinge linker of five amino acids. Remarkably, this short hinge plays a large role in the species-specific binding by the NS1B protein. The hinge of human and non-human primate ISG15, which has a sequence that differs from that of other mammalian ISG15 proteins, including mouse and canine ISG15, is absolutely required for binding the NS1B protein. Consequently, the ISG15 proteins of humans and non-human primates are the only mammalian ISG15 proteins that would bind NS1B.

  15. Dengue Virus NS1 Disrupts the Endothelial Glycocalyx, Leading to Hyperpermeability.

    PubMed

    Puerta-Guardo, Henry; Glasner, Dustin R; Harris, Eva

    2016-07-01

    Dengue is the most prevalent arboviral disease in humans and a major public health problem worldwide. Systemic plasma leakage, leading to hypovolemic shock and potentially fatal complications, is a critical determinant of dengue severity. Recently, we and others described a novel pathogenic effect of secreted dengue virus (DENV) non-structural protein 1 (NS1) in triggering hyperpermeability of human endothelial cells in vitro and systemic vascular leakage in vivo. NS1 was shown to activate toll-like receptor 4 signaling in primary human myeloid cells, leading to secretion of pro-inflammatory cytokines and vascular leakage. However, distinct endothelial cell-intrinsic mechanisms of NS1-induced hyperpermeability remained to be defined. The endothelial glycocalyx layer (EGL) is a network of membrane-bound proteoglycans and glycoproteins lining the vascular endothelium that plays a key role in regulating endothelial barrier function. Here, we demonstrate that DENV NS1 disrupts the EGL on human pulmonary microvascular endothelial cells, inducing degradation of sialic acid and shedding of heparan sulfate proteoglycans. This effect is mediated by NS1-induced expression of sialidases and heparanase, respectively. NS1 also activates cathepsin L, a lysosomal cysteine proteinase, in endothelial cells, which activates heparanase via enzymatic cleavage. Specific inhibitors of sialidases, heparanase, and cathepsin L prevent DENV NS1-induced EGL disruption and endothelial hyperpermeability. All of these effects are specific to NS1 from DENV1-4 and are not induced by NS1 from West Nile virus, a related flavivirus. Together, our data suggest an important role for EGL disruption in DENV NS1-mediated endothelial dysfunction during severe dengue disease. PMID:27416066

  16. Dengue Virus NS1 Disrupts the Endothelial Glycocalyx, Leading to Hyperpermeability

    PubMed Central

    Puerta-Guardo, Henry; Glasner, Dustin R.; Harris, Eva

    2016-01-01

    Dengue is the most prevalent arboviral disease in humans and a major public health problem worldwide. Systemic plasma leakage, leading to hypovolemic shock and potentially fatal complications, is a critical determinant of dengue severity. Recently, we and others described a novel pathogenic effect of secreted dengue virus (DENV) non-structural protein 1 (NS1) in triggering hyperpermeability of human endothelial cells in vitro and systemic vascular leakage in vivo. NS1 was shown to activate toll-like receptor 4 signaling in primary human myeloid cells, leading to secretion of pro-inflammatory cytokines and vascular leakage. However, distinct endothelial cell-intrinsic mechanisms of NS1-induced hyperpermeability remained to be defined. The endothelial glycocalyx layer (EGL) is a network of membrane-bound proteoglycans and glycoproteins lining the vascular endothelium that plays a key role in regulating endothelial barrier function. Here, we demonstrate that DENV NS1 disrupts the EGL on human pulmonary microvascular endothelial cells, inducing degradation of sialic acid and shedding of heparan sulfate proteoglycans. This effect is mediated by NS1-induced expression of sialidases and heparanase, respectively. NS1 also activates cathepsin L, a lysosomal cysteine proteinase, in endothelial cells, which activates heparanase via enzymatic cleavage. Specific inhibitors of sialidases, heparanase, and cathepsin L prevent DENV NS1-induced EGL disruption and endothelial hyperpermeability. All of these effects are specific to NS1 from DENV1-4 and are not induced by NS1 from West Nile virus, a related flavivirus. Together, our data suggest an important role for EGL disruption in DENV NS1-mediated endothelial dysfunction during severe dengue disease. PMID:27416066

  17. Raman spectroscopy based discrimination of NS1 positive and negative dengue virus infected serum

    NASA Astrophysics Data System (ADS)

    Bilal, M.; Saleem, M.; Bilal, Maria; Khurram, M.; Khan, Saranjam; Ullah, Rahat; Ali, Hina; Ahmed, M.

    2016-09-01

    This study is intended to develop a multivariate statistical model for the prediction of nonstructural protein 1 (NS1) in dengue virus (DENV) infected blood serum in humans. The model has been developed on the basis of partial least squares regression using the Raman spectra of NS1 positive and NS1 negative samples. Human blood sera of 218 subjects is included in this study, of which 95 were NS1 positive and 123 were NS1 negative, which was confirmed with the enzyme linked immunosorbent assay method. For model development, 80 NS1 positive and 98 NS1 negative samples were used, while 40 DENV suspected samples were used for double blind testing of the model. This selection of samples was performed by the code in an automatic manner to avoid biasing. A laser at 785 nm was used as the excitation source to acquire Raman spectra of samples with an integration time of 15 s. The multivariate model yields coefficients of regression at corresponding Raman shifts. These coefficients represent changes in the molecular structures associated with NS1 positive and negative samples. The analysis of the regression coefficients which differentiate NS1 positive and NS1 negative groups shows an increasing trend for phosphatidylinositol, ceramide, and amide-III, and a decreasing trend for thiocyanate in the DENV infected serum. The R-squared value of the model was found to be 0.91, which is clinically acceptable. The blind testing of 40 suspected samples yields an accuracy, sensitivity, and specificity of about 100% each.

  18. An observational correlation between stellar brightness variations and surface gravity.

    PubMed

    Bastien, Fabienne A; Stassun, Keivan G; Basri, Gibor; Pepper, Joshua

    2013-08-22

    Surface gravity is a basic stellar property, but it is difficult to measure accurately, with typical uncertainties of 25 to 50 per cent if measured spectroscopically and 90 to 150 per cent if measured photometrically. Asteroseismology measures gravity with an uncertainty of about 2 per cent but is restricted to relatively small samples of bright stars, most of which are giants. The availability of high-precision measurements of brightness variations for more than 150,000 stars provides an opportunity to investigate whether the variations can be used to determine surface gravities. The Fourier power of granulation on a star's surface correlates physically with surface gravity: if brightness variations on timescales of hours arise from granulation, then such variations should correlate with surface gravity. Here we report an analysis of archival data that reveals an observational correlation between surface gravity and root mean squared brightness variations on timescales of less than eight hours for stars with temperatures of 4,500 to 6,750 kelvin, log surface gravities of 2.5 to 4.5 (cgs units) and overall brightness variations of less than three parts per thousand. A straightforward observation of optical brightness variations therefore allows a determination of the surface gravity with a precision of better than 25 per cent for inactive Sun-like stars at main-sequence to giant stages of evolution. PMID:23969460

  19. The Influence of Microphysical Cloud Parameterization on Microwave Brightness Temperatures

    NASA Technical Reports Server (NTRS)

    Skofronick-Jackson, Gail M.; Gasiewski, Albin J.; Wang, James R.; Zukor, Dorothy J. (Technical Monitor)

    2000-01-01

    The microphysical parameterization of clouds and rain-cells plays a central role in atmospheric forward radiative transfer models used in calculating passive microwave brightness temperatures. The absorption and scattering properties of a hydrometeor-laden atmosphere are governed by particle phase, size distribution, aggregate density., shape, and dielectric constant. This study identifies the sensitivity of brightness temperatures with respect to the microphysical cloud parameterization. Cloud parameterizations for wideband (6-410 GHz observations of baseline brightness temperatures were studied for four evolutionary stages of an oceanic convective storm using a five-phase hydrometeor model in a planar-stratified scattering-based radiative transfer model. Five other microphysical cloud parameterizations were compared to the baseline calculations to evaluate brightness temperature sensitivity to gross changes in the hydrometeor size distributions and the ice-air-water ratios in the frozen or partly frozen phase. The comparison shows that, enlarging the rain drop size or adding water to the partly Frozen hydrometeor mix warms brightness temperatures by up to .55 K at 6 GHz. The cooling signature caused by ice scattering intensifies with increasing ice concentrations and at higher frequencies. An additional comparison to measured Convection and Moisture LA Experiment (CAMEX 3) brightness temperatures shows that in general all but, two parameterizations produce calculated T(sub B)'s that fall within the observed clear-air minima and maxima. The exceptions are for parameterizations that, enhance the scattering characteristics of frozen hydrometeors.

  20. An observational correlation between stellar brightness variations and surface gravity.

    PubMed

    Bastien, Fabienne A; Stassun, Keivan G; Basri, Gibor; Pepper, Joshua

    2013-08-22

    Surface gravity is a basic stellar property, but it is difficult to measure accurately, with typical uncertainties of 25 to 50 per cent if measured spectroscopically and 90 to 150 per cent if measured photometrically. Asteroseismology measures gravity with an uncertainty of about 2 per cent but is restricted to relatively small samples of bright stars, most of which are giants. The availability of high-precision measurements of brightness variations for more than 150,000 stars provides an opportunity to investigate whether the variations can be used to determine surface gravities. The Fourier power of granulation on a star's surface correlates physically with surface gravity: if brightness variations on timescales of hours arise from granulation, then such variations should correlate with surface gravity. Here we report an analysis of archival data that reveals an observational correlation between surface gravity and root mean squared brightness variations on timescales of less than eight hours for stars with temperatures of 4,500 to 6,750 kelvin, log surface gravities of 2.5 to 4.5 (cgs units) and overall brightness variations of less than three parts per thousand. A straightforward observation of optical brightness variations therefore allows a determination of the surface gravity with a precision of better than 25 per cent for inactive Sun-like stars at main-sequence to giant stages of evolution.

  1. Global View of the Bright Material on Vesta

    NASA Technical Reports Server (NTRS)

    Zambon, F.; DeSanctis, C.; Schroeder, S.; Tosi, F.; Li, J.-Y.; Longobardo, A.; Ammannito, E.; Blewett, D. T.; Palomba, E.; Capaccioni, F.; Frigeri, A.; Capria, M. T.; Fonte, S.; Mittlefehldt, D. W.; Nathues, A.; Pieters, C.; Russell, C. T.; Raymond, C. A.

    2014-01-01

    At 525 km in mean diameter, Vesta is the second-most massive and one of the brightest asteroids of the main-belt. Here we give a global view of the bright material (BM) units on Vesta. We classified the BMs according to the normal visual albedo. The global albedo map of Vesta allows to be divided the surface into three principal types of terrains: bright regions, dark regions and intermediate regions. The distribution of bright regions is not uniform. The mid-southern latitudes contain the most bright areas, while the northern hemisphere is poor in bright regions. The analysis of the spectral parameters and the normal visual albedo show a dependence between albedo and the strength (depth) of ferrous iron absorption bands, strong bands correspond with high albedo units. Vesta's average albedo is 0.38, but there are bright material whose albedo can exceed 0.50. Only the E-Type asteroids have albedos comparable to those of the BMs on Vesta. The Dawn mission observed a large fraction of Vesta's surface at high spatial resolution, allowing a detailed study of the morphology and mineralogy of it. In particular, reflectance spectra provided by the Visible and InfraRed spectrometer (VIR), confirmed that Vesta's mineralogy is dominated by pyroxenes. All Vesta spectra show two strong absorption bands at approx 0.9 and 1.9 micron, typical of the pyroxenes and associated with the howardite, eucrite and diogenite (HED) meteorites.

  2. Synthesizing SMOS Zero-Baselines with Aquarius Brightness Temperature Simulator

    NASA Technical Reports Server (NTRS)

    Colliander, A.; Dinnat, E.; Le Vine, D.; Kainulainen, J.

    2012-01-01

    SMOS [1] and Aquarius [2] are ESA and NASA missions, respectively, to make L-band measurements from the Low Earth Orbit. SMOS makes passive measurements whereas Aquarius measures both passive and active. SMOS was launched in November 2009 and Aquarius in June 2011.The scientific objectives of the missions are overlapping: both missions aim at mapping the global Sea Surface Salinity (SSS). Additionally, SMOS mission produces soil moisture product (however, Aquarius data will eventually be used for retrieving soil moisture too). The consistency of the brightness temperature observations made by the two instruments is essential for long-term studies of SSS and soil moisture. For resolving the consistency, the calibration of the instruments is the key. The basis of the SMOS brightness temperature level is the measurements performed with the so-called zero-baselines [3]; SMOS employs an interferometric measurement technique which forms a brightness temperature image from several baselines constructed by combination of multiple receivers in an array; zero-length baseline defines the overall brightness temperature level. The basis of the Aquarius brightness temperature level is resolved from the brightness temperature simulator combined with ancillary data such as antenna patterns and environmental models [4]. Consistency between the SMOS zero-baseline measurements and the simulator output would provide a robust basis for establishing the overall comparability of the missions.

  3. NMR and MD Studies Reveal That the Isolated Dengue NS3 Protease Is an Intrinsically Disordered Chymotrypsin Fold Which Absolutely Requests NS2B for Correct Folding and Functional Dynamics

    PubMed Central

    Gupta, Garvita; Lim, Liangzhong; Song, Jianxing

    2015-01-01

    Dengue genome encodes a two component protease complex (NS2B-NS3pro) essential for the viral maturation/infectivity, thus representing a key drug target. Previously, due to its “complete insolubility”, the isolated NS3pro could not be experimentally studied and it remains elusive what structure it adopts without NS2B and why NS2B is indispensable. Here as facilitated by our previous discovery, the isolated NS3pro has been surprisingly deciphered by NMR to be the first intrinsically-disordered chymotrypsin-like fold, which exists in a loosely-packed state with non-native long-range interactions as revealed by paramagnetic relaxation enhancement (PRE). The disordered NS3pro appears to be needed for binding a human host factor to trigger the membrane remodeling. Moreover, we have in vitro refolded the NS3pro in complex with either NS2B (48–100) or the full-length NS2B (1–130) anchored into the LMPC micelle, and the two complexes have similar activities but different dynamics. We also performed molecular dynamics (MD) simulations and the results revealed that NS2B shows the highest structural fluctuations in the complex, thus providing the dynamic basis for the observation on its conformational exchange between open and closed states. Remarkably, the NS2B cofactor plays a central role in maintaining the correlated motion network required for the catalysis as we previously decoded for the SARS 3CL protease. Indeed, a truncated NS2B (48–100;Δ77–84) with the flexible loop deleted is able to trap the NS2B-NS3pro complex in a highly dynamic and catalytically-impotent state. Taken together, our study implies potential strategies to perturb the NS2B-NS3pro interface for design of inhibitors for treating dengue infection. PMID:26258523

  4. High peak- and average-power pulse shaped fiber laser in the ns-regime applying step-index XLMA gain fibers

    NASA Astrophysics Data System (ADS)

    Dinger, R.; Grundmann, F.-P.; Hapke, C.; Ruppik, S.

    2014-03-01

    Pulsed fiber lasers and continuous-wave (cw) fiber lasers have become the tool of choice in more and more laser based industrial applications like metal cutting and welding mainly because of their robustness, compactness, high brightness, high efficiency and reasonable costs. However, to further increase the productivity with those laser types there is a great demand for even higher laser power specifications. In this context we demonstrate a pulsed high peak- and averagepower fiber laser in a Master Oscillator Power Amplifier (MOPA) configuration with selectable pulse durations between 1 ns and several hundred nanoseconds. To overcome fiber nonlinearities such as stimulated Raman scattering (SRS) and self-phase-modulation (SPM) flexible Ytterbium doped extra-large mode area (XLMA) step index fibers, prepared by novel powder-sinter technology, have been used as gain fibers. As an example, for 12 ns pulses with a repetition rate of 10 kHz, a pump power limited average laser output power of more than 400 W in combination with peak powers of more than 3.5 MW (close to self-focusing-threshold) has been achieved in stable operation. The potentials of this laser system have been further explored towards longer pulse durations in order to achieve even higher pulse energies by means of pulse shaping techniques. In addition, investigations have been conducted with reduced pulse energies and repetition rates up to 500 kHz and average powers of more than 500 W at nearly diffraction limited beam quality.

  5. Epitope mapping and functional analysis of sigma A and sigma NS proteins of avian reovirus

    SciTech Connect

    Huang, Pi H.; Li, Ying J.; Su, Yu P.; Lee, Long H.; Liu, Hung J. . E-mail: hjliu@mail.npust.edu.tw

    2005-02-20

    We have previously shown that avian reovirus (ARV) {sigma}A and {sigma}NS proteins possess dsRNA and ssRNA binding activity and suggested that there are two epitopes on {sigma}A (I and II) and three epitopes (A, B, and C) on {sigma}NS. To further define the location of epitopes on {sigma}A and {sigma}NS proteins and to further elucidate the biological functions of these epitopes by using monoclonal antibodies (MAbs) 62, 1F9, H1E1, and 4A123 against the ARV S1133 strain, the full-length and deletion fragments of S2 and S4 genes of ARV generated by polymerase chain reaction (PCR) were cloned into pET32 expression vectors and the fusion proteins were overexpressed in Escherichia coli BL21 strain. Epitope mapping using MAbs and E. coli-expressed deletion fragments of {sigma}A and {sigma}NS of the ARV S1133 strain, synthetic peptides, and the cross reactivity of MAbs to heterologous ARV strains demonstrated that epitope II on {sigma}A was located at amino acid residues {sup 340}QWVMAGLVSAA{sup 350} and epitope B on {sigma}NS at amino acid residues {sup 180}MLDMVDGRP{sup 188}. The MAbs (62, 1F9, and H1E1) directed against epitopes II and B did not require the native conformation of {sigma}A and {sigma}NS, suggesting that their binding activities were conformation-independent. On the other hand, MAb 4A123 only reacted with complete {sigma}NS but not with truncated {sigma}NS fusion proteins in Western blot, suggesting that the binding activity of MAb to epitope A on {sigma}NS was conformation-dependent. Amino acid sequence analysis and the binding assays of MAb 62 to heterologous ARV strains suggested that epitope II on {sigma}A was highly conserved among ARV strains and that this epitope is suitable as a serological marker for the detection of ARV antibodies following natural infection in chickens. On the contrary, an amino acid substitution at position 183 (M to V) in epitope B of ARV could hinder the reactivity of the {sigma}NS with MAb 1F9. The {sigma}NS of ARV with ss

  6. Construction and analysis of a plant non-specific lipid transfer protein database (nsLTPDB)

    PubMed Central

    2012-01-01

    Background Plant non-specific lipid transfer proteins (nsLTPs) are small and basic proteins. Recently, nsLTPs have been reported involved in many physiological functions such as mediating phospholipid transfer, participating in plant defence activity against bacterial and fungal pathogens, and enhancing cell wall extension in tobacco. However, the lipid transfer mechanism of nsLTPs is still unclear, and comprehensive information of nsLTPs is difficult to obtain. Methods In this study, we identified 595 nsLTPs from 121 different species and constructed an nsLTPs database -- nsLTPDB -- which comprises the sequence information, structures, relevant literatures, and biological data of all plant nsLTPs http://nsltpdb.life.nthu.edu.tw/. Results Meanwhile, bioinformatics and statistics methods were implemented to develop a classification method for nsLTPs based on the patterns of the eight highly-conserved cysteine residues, and to suggest strict Prosite-styled patterns for Type I and Type II nsLTPs. The pattern of Type I is C X2 V X5-7 C [V, L, I] × Y [L, A, V] X8-13 CC × G X12 D × [Q, K, R] X2 CXC X16-21 P X2 C X13-15C, and that of Type II is C X4 L X2 C X9-11 P [S, T] X2 CC X5 Q X2-4 C[L, F]C X2 [A, L, I] × [D, N] P X10-12 [K, R] X4-5 C X3-4 P X0-2 C. Moreover, we referred the Prosite-styled patterns to the experimental mutagenesis data that previously established by our group, and found that the residues with higher conservation played an important role in the structural stability or lipid binding ability of nsLTPs. Conclusions Taken together, this research has suggested potential residues that might be essential to modulate the structural and functional properties of plant nsLTPs. Finally, we proposed some biologically important sites of the nsLTPs, which are described by using a new Prosite-styled pattern that we defined. PMID:22369214

  7. X-Ray Structure of the Pestivirus NS3 Helicase and Its Conformation in Solution

    PubMed Central

    Duquerroy, Stéphane; Kwok, Jane; Vonrhein, Clemens; Perez, Javier; Lamp, Benjamin; Bricogne, Gerard; Rümenapf, Till; Vachette, Patrice

    2015-01-01

    ABSTRACT Pestiviruses form a genus in the Flaviviridae family of small enveloped viruses with a positive-sense single-stranded RNA genome. Viral replication in this family requires the activity of a superfamily 2 RNA helicase contained in the C-terminal domain of nonstructural protein 3 (NS3). NS3 features two conserved RecA-like domains (D1 and D2) with ATPase activity, plus a third domain (D3) that is important for unwinding nucleic acid duplexes. We report here the X-ray structure of the pestivirus NS3 helicase domain (pNS3h) at a 2.5-Å resolution. The structure deviates significantly from that of NS3 of other genera in the Flaviviridae family in D3, as it contains two important insertions that result in a narrower nucleic acid binding groove. We also show that mutations in pNS3h that rescue viruses from which the core protein is deleted map to D3, suggesting that this domain may be involved in interactions that facilitate particle assembly. Finally, structural comparisons of the enzyme in different crystalline environments, together with the findings of small-angle X-ray-scattering studies in solution, show that D2 is mobile with respect to the rest of the enzyme, oscillating between closed and open conformations. Binding of a nonhydrolyzable ATP analog locks pNS3h in a conformation that is more compact than the closest apo-form in our crystals. Together, our results provide new insight and bring up new questions about pNS3h function during pestivirus replication. IMPORTANCE Although pestivirus infections impose an important toll on the livestock industry worldwide, little information is available about the nonstructural proteins essential for viral replication, such as the NS3 helicase. We provide here a comparative structural and functional analysis of pNS3h with respect to its orthologs in other viruses of the same family, the flaviviruses and hepatitis C virus. Our studies reveal differences in the nucleic acid binding groove that could have implications

  8. Population pharmacokinetic modelling of NS2330 (tesofensine) and its major metabolite in patients with Alzheimer's disease

    PubMed Central

    Lehr, Thorsten; Staab, Alexander; Tillmann, Christiane; Trommeshauser, Dirk; Raschig, Andreas; Schaefer, Hans Guenter; Kloft, Charlotte

    2007-01-01

    What is already known about this subject Several studies in predominantly healthy subjects have investigated the pharmacokinetics of NS2330 and its major metabolite M1. However, its pharmacokinetics have not been characterized in Alzheimer's disease patients, the target population for NS2330. In addition, no covariates have previously been found to influence the plasma concentration-time profiles of NS2330 and/or M1. What this study adds A descriptive and predictive population pharmacokinetic model for NS2330 and its metabolite was successfully developed in a population of patients with Alzheimer's disease. A covariate analysis elucidated sex and creatinine clearance as having an influence on the plasma concentration-time profiles of NS2330 after long-term treatment. Aims To develop a population pharmacokinetic model for NS2330 and its major metabolite M1 based on data from a 14 week proof of concept study in patients with Alzheimer's disease, and to identify covariates that might influence the pharmacokinetic characteristics of the drug and/or its metabolite. Methods Plasma data from 320 subjects undergoing multiple oral dosing, and consisting of 1969 NS2330 and 1714 metabolite concentrations were fitted simultaneously using NONMEM. Results Plasma concentration-time profiles of NS2330 and M1 were best described by one-compartment models with first-order elimination for both compounds. Absorption of NS2330 was best modelled by a first-order process. Low apparent clearances together with large apparent volumes of distribution resulted in long half-lives of 234 h (NS2330) and 374 h (M1). The covariate analysis identified weight, sex, CLCR, BMI and age as influencing the pharmacokinetics of NS2330 and/or M1. However, simulations performed revealed that only CLCR and sex had a significant effect on the steady-state plasma concentration-time profiles. Females with a creatinine clearance of 35.6 ml min−1 showed a 62% increased exposure compared with males without renal

  9. Influenza virus NS1 protein inhibits pre-mRNA splicing and blocks mRNA nucleocytoplasmic transport.

    PubMed

    Fortes, P; Beloso, A; Ortín, J

    1994-02-01

    The influenza virus RNA segment 8 encodes two proteins, NS1 and NS2, by differential splicing. The collinear transcript acts as mRNA for NS1 protein, while the spliced mRNA encodes NS2 protein. The splicing of NS1 mRNA was studied in cells transfected with a recombinant plasmid that has the cDNA of RNA segment 8 cloned under the SV40 late promoter and polyadenylation signals. As described for influenza virus-infected cells, NS1 mRNA was poorly spliced to yield NS2 mRNA. However, inactivation of the NS1 gene, but not the NS2 gene, led to a substantial increase in the splicing efficiency, as shown by the relative accumulations of NS1 and NS2 mRNAs. This effect was not specific for NS1 mRNA, since the splicing of the endogenous SV40 early transcript was altered in such a way that t-Ag mRNA was almost eliminated. These changes in the splicing pattern coincided with a strong inhibition of the mRNA nucleocytoplasmic transport. Both NS1 and NS2 mRNAs were retained in the nucleus of cells expressing NS1 protein, but no effect was observed when only NS2 protein was expressed. Furthermore, other mRNAs tested, such as T-Ag mRNA and the non-spliceable nucleoprotein transcript, were also retained in the nucleus upon expression of NS1 protein, suggesting that it induced a generalized block of mRNA export from the nucleus.

  10. The N-Terminal of Aquareovirus NS80 Is Required for Interacting with Viral Proteins and Viral Replication

    PubMed Central

    Zhang, Jie; Guo, Hong; Chen, Qingxiu; Zhang, Fuxian; Fang, Qin

    2016-01-01

    Reovirus replication and assembly occurs within viral inclusion bodies that formed in specific intracellular compartments of cytoplasm in infected cells. Previous study indicated that aquareovirus NS80 is able to form inclusion bodies, and also can retain viral proteins within its inclusions. To better understand how NS80 performed in viral replication and assembly, the functional regions of NS80 associated with other viral proteins in aquareovirus replication were investigated in this study. Deletion mutational analysis and rotavirus NSP5-based protein association platform were used to detect association regions. Immunofluorescence images indicated that different N-terminal regions of NS80 could associate with viral proteins VP1, VP4, VP6 and NS38. Further co-immunoprecipitation analysis confirmed the interaction between VP1, VP4, VP6 or NS38 with different regions covering the N-terminal amino acid (aa, 1–471) of NS80, respectively. Moreover, removal of NS80 N-terminal sequences required for interaction with proteins VP1, VP4, VP6 or NS38 not only prevented the capacity of NS80 to support viral replication in NS80 shRNA-based replication complementation assays, but also inhibited the expression of aquareovirus proteins, suggesting that N-terminal regions of NS80 are necessary for viral replication. These results provided a foundational basis for further understanding the role of NS80 in viral replication and assembly during aquareovirus infection. PMID:26871941

  11. Molecular Modeling and Docking Study to Elucidate Novel Chikungunya Virus nsP2 Protease Inhibitors

    PubMed Central

    Agarwal, T.; Asthana, Somya; Bissoyi, A.

    2015-01-01

    Chikungunya is one of the tropical viral infections that severely affect the Asian and African countries. Absence of any suitable drugs or vaccines against Chikungunya virus till date makes it essential to identify and develop novel leads for the same. Recently, nsP2 cysteine protease has been classified as a crucial drug target to combat infections caused by Alphaviruses including Chikungunya virus due to its involvement viral replication. Here in, we investigated the structural aspects of the nsP2 protease through homology modeling based on nsP2 protease from Venezuelan equine encephalitis virus. Further, the ligands were virtually screened based on various pharmacological, ADME/Tox filters and subjected to docking with the modeled Chikungunya nsP2 protease using AutoDock4.2. The interaction profiling of ligand with the protein was carried out using LigPlot+. The results demonstrated that the ligand with PubChem Id (CID_5808891) possessed highest binding affinity towards Chikungunya nsP2 protease with a good interaction profile with the active site residues. We hereby propose that these compounds could inhibit the nsP2 protease by binding to its active site. Moreover, they may provide structural scaffold for the design of novel leads with better efficacy and specificity for the nsP2 protease. PMID:26664062

  12. Increased Bending Rigidity of Single DNA Molecules by H-NS, a Temperature and Osmolarity Sensor

    PubMed Central

    Amit, Roee; Oppenheim, Amos B.; Stavans, Joel

    2003-01-01

    Histonelike nucleoid structuring protein (H-NS) is an abundant prokaryotic protein participating in nucleoid structure, gene regulation, and silencing. It plays a key role in cell response to changes in temperature and osmolarity. Force-extension measurements of single, twist-relaxed λ-DNA-H-NS complexes show that these adopt more extended configurations compared to the naked DNA substrates. Crosslinking indicates that H-NS can decorate DNA molecules at one H-NS dimer per 15–20 bp. These results suggest that H-NS polymerizes along DNA, forming a complex of higher bending rigidity. These effects are not observed above 32°C or at high osmolarity, supporting the hypothesis that a direct H-NS-DNA interaction plays a key role in gene silencing. Thus, we propose that H-NS plays a unique structural role, different from that of HU and IHF, and functions as one of the environmental sensors of the cell. PMID:12668454

  13. The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing.

    PubMed

    De Maio, Federico A; Risso, Guillermo; Iglesias, Nestor G; Shah, Priya; Pozzi, Berta; Gebhard, Leopoldo G; Mammi, Pablo; Mancini, Estefania; Yanovsky, Marcelo J; Andino, Raul; Krogan, Nevan; Srebrow, Anabella; Gamarnik, Andrea V

    2016-08-01

    Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of infected cells together with functional studies, we found that NS5 binds spliceosome complexes and modulates endogenous splicing as well as minigene-derived alternative splicing patterns. In particular, we show that NS5 alone, or in the context of viral infection, interacts with core components of the U5 snRNP particle, CD2BP2 and DDX23, alters the inclusion/exclusion ratio of alternative splicing events, and changes mRNA isoform abundance of known antiviral factors. Interestingly, a genome wide transcriptome analysis, using recently developed bioinformatics tools, revealed an increase of intron retention upon dengue virus infection, and viral replication was improved by silencing specific U5 components. Different mechanistic studies indicate that binding of NS5 to the spliceosome reduces the efficiency of pre-mRNA processing, independently of NS5 enzymatic activities. We propose that NS5 binding to U5 snRNP proteins hijacks the splicing machinery resulting in a less restrictive environment for viral replication. PMID:27575636

  14. Raising the avermectins production in Streptomyces avermitilis by utilizing nanosecond pulsed electric fields (nsPEFs)

    NASA Astrophysics Data System (ADS)

    Guo, Jinsong; Ma, Ruonan; Su, Bo; Li, Yinglong; Zhang, Jue; Fang, Jing

    2016-05-01

    Avermectins, a group of anthelmintic and insecticidal agents produced from Streptomyces avermitilis, are widely used in agricultural, veterinary, and medical fields. This study presents the first report on the potential of using nanosecond pulsed electric fields (nsPEFs) to improve avermectin production in S. avermitilis. The results of colony forming units showed that 20 pulses of nsPEFs at 10 kV/cm and 20 kV/cm had a significant effect on proliferation, while 100 pulses of nsPEFs at 30 kV/cm exhibited an obvious effect on inhibition of agents. Ultraviolet spectrophotometry assay revealed that 20 pulses of nsPEFs at 15 kV/cm increased avermectin production by 42% and reduced the time for reaching a plateau in fermentation process from 7 days to 5 days. In addition, the decreased oxidation reduction potential (ORP) and increased temperature of nsPEFs-treated liquid were evidenced to be closely associated with the improved cell growth and fermentation efficiency of avermectins in S. avermitilis. More importantly, the real-time RT-PCR analysis showed that nsPEFs could remarkably enhance the expression of aveR and malE in S. avermitilis during fermentation, which are positive regulator for avermectin biosynthesis. Therefore, the nsPEFs technology presents an alternative strategy to be developed to increase avermectin output in fermentation industry.

  15. The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing.

    PubMed

    De Maio, Federico A; Risso, Guillermo; Iglesias, Nestor G; Shah, Priya; Pozzi, Berta; Gebhard, Leopoldo G; Mammi, Pablo; Mancini, Estefania; Yanovsky, Marcelo J; Andino, Raul; Krogan, Nevan; Srebrow, Anabella; Gamarnik, Andrea V

    2016-08-01

    Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of infected cells together with functional studies, we found that NS5 binds spliceosome complexes and modulates endogenous splicing as well as minigene-derived alternative splicing patterns. In particular, we show that NS5 alone, or in the context of viral infection, interacts with core components of the U5 snRNP particle, CD2BP2 and DDX23, alters the inclusion/exclusion ratio of alternative splicing events, and changes mRNA isoform abundance of known antiviral factors. Interestingly, a genome wide transcriptome analysis, using recently developed bioinformatics tools, revealed an increase of intron retention upon dengue virus infection, and viral replication was improved by silencing specific U5 components. Different mechanistic studies indicate that binding of NS5 to the spliceosome reduces the efficiency of pre-mRNA processing, independently of NS5 enzymatic activities. We propose that NS5 binding to U5 snRNP proteins hijacks the splicing machinery resulting in a less restrictive environment for viral replication.

  16. Thermodynamics of zinc binding to hepatitis C virus NS3 protease: a folding by binding event.

    PubMed

    Abian, Olga; Neira, Jose Luis; Velazquez-Campoy, Adrian

    2009-11-15

    The hepatitis C virus (HCV) nonstructural protein 3 (NS3) protease is responsible for the processing of the non-structural region of the viral precursor polyprotein in infected hepatic cells. HCV NS3 is a zinc-dependent serine protease. The zinc ion, which is bound far away from the active site and considered to have a structural role, is essential for the structural integrity of the protein; furthermore, the ion is required for the hydrolytic activity. Consequently, the NS3 zinc binding site has been considered for a long time as a possible target for drug discovery. As a first step towards this goal, the energetics of the NS3-zinc interaction and its effect on the NS3 conformation must be established and discussed. The thermodynamic characterization of zinc binding to NS3 protease by isothermal titration calorimetry and spectroscopy is presented here. Spectroscopic and calorimetric results suggest that a considerable conformational change in the protein is coupled to zinc binding. The energetics of the conformational change is comparable to that of the folding of a protein of similar size. Therefore, zinc binding to NS3 protease can be considered as a "folding by binding" event.

  17. The Dengue Virus NS5 Protein Intrudes in the Cellular Spliceosome and Modulates Splicing

    PubMed Central

    Shah, Priya; Pozzi, Berta; Gebhard, Leopoldo G.; Mammi, Pablo; Yanovsky, Marcelo J.; Andino, Raul; Krogan, Nevan; Srebrow, Anabella; Gamarnik, Andrea V.

    2016-01-01

    Dengue virus NS5 protein plays multiple functions in the cytoplasm of infected cells, enabling viral RNA replication and counteracting host antiviral responses. Here, we demonstrate a novel function of NS5 in the nucleus where it interferes with cellular splicing. Using global proteomic analysis of infected cells together with functional studies, we found that NS5 binds spliceosome complexes and modulates endogenous splicing as well as minigene-derived alternative splicing patterns. In particular, we show that NS5 alone, or in the context of viral infection, interacts with core components of the U5 snRNP particle, CD2BP2 and DDX23, alters the inclusion/exclusion ratio of alternative splicing events, and changes mRNA isoform abundance of known antiviral factors. Interestingly, a genome wide transcriptome analysis, using recently developed bioinformatics tools, revealed an increase of intron retention upon dengue virus infection, and viral replication was improved by silencing specific U5 components. Different mechanistic studies indicate that binding of NS5 to the spliceosome reduces the efficiency of pre-mRNA processing, independently of NS5 enzymatic activities. We propose that NS5 binding to U5 snRNP proteins hijacks the splicing machinery resulting in a less restrictive environment for viral replication. PMID:27575636

  18. Identification of Hydroxyanthraquinones as Novel Inhibitors of Hepatitis C Virus NS3 Helicase

    PubMed Central

    Furuta, Atsushi; Tsubuki, Masayoshi; Endoh, Miduki; Miyamoto, Tatsuki; Tanaka, Junichi; Abdus Salam, Kazi; Akimitsu, Nobuyoshi; Tani, Hidenori; Yamashita, Atsuya; Moriishi, Kohji; Nakakoshi, Masamichi; Sekiguchi, Yuji; Tsuneda, Satoshi; Noda, Naohiro

    2015-01-01

    Hepatitis C virus (HCV) is an important etiological agent of severe liver diseases, including cirrhosis and hepatocellular carcinoma. The HCV genome encodes nonstructural protein 3 (NS3) helicase, which is a potential anti-HCV drug target because its enzymatic activity is essential for viral replication. Some anthracyclines are known to be NS3 helicase inhibitors and have a hydroxyanthraquinone moiety in their structures; mitoxantrone, a hydroxyanthraquinone analogue, is also known to inhibit NS3 helicase. Therefore, we hypothesized that the hydroxyanthraquinone moiety alone could also inhibit NS3 helicase. Here, we performed a structure–activity relationship study on a series of hydroxyanthraquinones by using a fluorescence-based helicase assay. Hydroxyanthraquinones inhibited NS3 helicase with IC50 values in the micromolar range. The inhibitory activity varied depending on the number and position of the phenolic hydroxyl groups, and among different hydroxyanthraquinones examined, 1,4,5,8-tetrahydroxyanthraquinone strongly inhibited NS3 helicase with an IC50 value of 6 µM. Furthermore, hypericin and sennidin A, which both have two hydroxyanthraquinone-like moieties, were found to exert even stronger inhibition with IC50 values of 3 and 0.8 µM, respectively. These results indicate that the hydroxyanthraquinone moiety can inhibit NS3 helicase and suggest that several key chemical structures are important for the inhibition. PMID:26262613

  19. Raising the avermectins production in Streptomyces avermitilis by utilizing nanosecond pulsed electric fields (nsPEFs)

    PubMed Central

    Guo, Jinsong; Ma, Ruonan; Su, Bo; Li, Yinglong; Zhang, Jue; Fang, Jing

    2016-01-01

    Avermectins, a group of anthelmintic and insecticidal agents produced from Streptomyces avermitilis, are widely used in agricultural, veterinary, and medical fields. This study presents the first report on the potential of using nanosecond pulsed electric fields (nsPEFs) to improve avermectin production in S. avermitilis. The results of colony forming units showed that 20 pulses of nsPEFs at 10 kV/cm and 20 kV/cm had a significant effect on proliferation, while 100 pulses of nsPEFs at 30 kV/cm exhibited an obvious effect on inhibition of agents. Ultraviolet spectrophotometry assay revealed that 20 pulses of nsPEFs at 15 kV/cm increased avermectin production by 42% and reduced the time for reaching a plateau in fermentation process from 7 days to 5 days. In addition, the decreased oxidation reduction potential (ORP) and increased temperature of nsPEFs-treated liquid were evidenced to be closely associated with the improved cell growth and fermentation efficiency of avermectins in S. avermitilis. More importantly, the real-time RT-PCR analysis showed that nsPEFs could remarkably enhance the expression of aveR and malE in S. avermitilis during fermentation, which are positive regulator for avermectin biosynthesis. Therefore, the nsPEFs technology presents an alternative strategy to be developed to increase avermectin output in fermentation industry. PMID:27181521

  20. Antagonism of the complement component C4 by flavivirus nonstructural protein NS1

    PubMed Central

    Avirutnan, Panisadee; Fuchs, Anja; Hauhart, Richard E.; Somnuke, Pawit; Youn, Soonjeon

    2010-01-01

    The complement system plays an essential protective role in the initial defense against many microorganisms. Flavivirus NS1 is a secreted nonstructural glycoprotein that accumulates in blood, is displayed on the surface of infected cells, and has been hypothesized to have immune evasion functions. Herein, we demonstrate that dengue virus (DENV), West Nile virus (WNV), and yellow fever virus (YFV) NS1 attenuate classical and lectin pathway activation by directly interacting with C4. Binding of NS1 to C4 reduced C4b deposition and C3 convertase (C4b2a) activity. Although NS1 bound C4b, it lacked intrinsic cofactor activity to degrade C4b, and did not block C3 convertase formation or accelerate decay of the C3 and C5 convertases. Instead, NS1 enhanced C4 cleavage by recruiting and activating the complement-specific protease C1s. By binding C1s and C4 in a complex, NS1 promotes efficient degradation of C4 to C4b. Through this mechanism, NS1 protects DENV from complement-dependent neutralization in solution. These studies define a novel immune evasion mechanism for restricting complement control of microbial infection. PMID:20308361

  1. Structure of the catalytic domain of the hepatitis C virus NS2-3 protease

    SciTech Connect

    Lorenz,I.; Marcotrigiano, J.; Dentzer, T.; Rice, C.

    2006-01-01

    Hepatitis C virus is a major global health problem affecting an estimated 170 million people worldwide. Chronic infection is common and can lead to cirrhosis and liver cancer. There is no vaccine available and current therapies have met with limited success. The viral RNA genome encodes a polyprotein that includes two proteases essential for virus replication. The NS2-3 protease mediates a single cleavage at the NS2/NS3 junction, whereas the NS3-4A protease cleaves at four downstream sites in the polyprotein. NS3-4A is characterized as a serine protease with a chymotrypsin-like fold, but the enzymatic mechanism of the NS2-3 protease remains unresolved. Here we report the crystal structure of the catalytic domain of the NS2-3 protease at 2.3 Angstroms resolution. The structure reveals a dimeric cysteine protease with two composite active sites. For each active site, the catalytic histidine and glutamate residues are contributed by one monomer, and the nucleophilic cysteine by the other. The carboxy-terminal residues remain coordinated in the two active sites, predicting an inactive post-cleavage form. Proteolysis through formation of a composite active site occurs in the context of the viral polyprotein expressed in mammalian cells. These features offer unexpected insights into polyprotein processing by hepatitis C virus and new opportunities for antiviral drug design.

  2. Heterologous production, purification and characterization of enzymatically active Sindbis virus nonstructural protein nsP1.

    PubMed

    Tomar, Shailly; Narwal, Manju; Harms, Etti; Smith, Janet L; Kuhn, Richard J

    2011-10-01

    Alphavirus nonstructural protein nsP1 possesses distinct methyltransferase (MTase) and guanylyltransferase (GTase) activities involved in the capping of viral RNAs. In alphaviruses, the methylation of GTP occurs before RNA transguanylation and nsP1 forms a covalent complex with m(7)GMP unlike the host mRNA guanylyltransferase which forms GMP-enzyme complex. In this study, full length SINV nsP1 was expressed in a soluble form with an N-terminal histidine tag in Escherichia coli and purified to homogeneity. The purified protein is enzymatically active and contains both MTase and GTase activity indicating that SINV nsP1 does not require membrane association for its enzymatic function. Biochemical analysis shows that detergents abolish nsP1 GTase activity, whereas nonionic detergents do not affect MTase activity. Furthermore, SINV nsP1 contains the metal-ion dependent GTase, whereas MTase does not require a metal ion. Circular dichroism spectroscopic analysis of purified protein indicate that nsP1 has a mixed α/β structure and is in the folded native conformation. PMID:21693190

  3. Selection of RNA aptamers that bind specifically to the NS3 protease of hepatitis C virus.

    PubMed

    Urvil, P T; Kakiuchi, N; Zhou, D M; Shimotohno, K; Kumar, P K; Nishikawa, S

    1997-08-15

    The RNA genome of human hepatitis C virus (HCV) is translated into a large precursor polyprotein. The NS3 protease of HCV has a crucial role in the processing of the polyprotein into functional viral proteins. We have used an in vitro genetic-selection strategy to isolate high-affinity RNA aptamers that bind to the NS3 protein, especially to its protease domain. Starting from a RNA pool that had a random sequence core of 12-18 nucleotides, aptamers that bind specifically to the NS3 protein were selected after 10 rounds of selection and amplification. A single aptamer, 10G-1, was found predominantly (71%) in the selected pool. This aptamer could bind to the NS3 protein with a binding constant of 650 nM and inhibit the proteolytic activity in vitro. By phosphate-modification-interference analysis we showed that the phosphate residues that are critical for the binding of 10G-1 to NS3 lie within the selected regions of the aptamer and that binding involves electrostatic contacts with the phosphates of regions G28-U34 and A47-A55. The NS3-binding region in 10G-1 can serve as a basis for designing more potential inhibitors of the NS3 protein.

  4. [Preparation of reactive bright blue praseodymium dyestuff and its spectral properties].

    PubMed

    Liu, Xiao-Zhen; Yuan, Ya-Qin; Cai, Yu; Zhu, Xian; Wang, Yan-Hong

    2004-08-01

    Reactive bright blue praseodymium dyestuff was prepared by using reactive bright blue and praseodymium oxide. The spectra of reactive bright blue praseodymium and dyed silk cloth by reactive bright blue praseodymium dyestuff were studied by UV-Vis and IR spectra respectively. In the range of 200-800 nm, reactive bright blue has four absorption peaks, and lambda(max) is 259 nm; reactive bright blue praseodymium has three absorption peaks, while lambda(max), is 264.00 nm. In the range of 420-760 nm, reactive bright blue has two absorption peaks at 661.50 and 625.50 nm, respectively, and lambda(max) is 661.50 nm; reactive bright blue praseodymium has only one absorption peak at 618.00 nm. Coordinate bond links reactive bright blue to praseodymium ion. Reactive bright blue praseodymium increases linking radicals as compared with reactive bright blue.

  5. Dynamic Imaging of the Hepatitis C Virus NS5A Protein during a Productive Infection

    PubMed Central

    Eyre, Nicholas S.; Fiches, Guillaume N.; Aloia, Amanda L.; Helbig, Karla J.; McCartney, Erin M.; McErlean, Christopher S. P.; Li, Kui; Aggarwal, Anupriya; Turville, Stuart G.

    2014-01-01

    ABSTRACT Hepatitis C virus (HCV) NS5A is essential for viral genome replication within cytoplasmic replication complexes and virus assembly at the lipid droplet (LD) surface, although its definitive functions are poorly understood. We developed approaches to investigate NS5A dynamics during a productive infection. We report here that NS5A motility and efficient HCV RNA replication require the microtubule network and the cytoplasmic motor dynein and demonstrate that both motile and relatively static NS5A-positive foci are enriched with host factors VAP-A and Rab5A. Pulse-chase imaging revealed that newly synthesized NS5A foci are small and distinct from aged foci, while further studies using a unique dual fluorescently tagged infectious HCV chimera showed a relatively stable association of NS5A foci with core-capped LDs. These results reveal new details about the dynamics and maturation of NS5A and the nature of potential sites of convergence of HCV replication and assembly pathways. IMPORTANCE Hepatitis C virus (HCV) is a major cause of serious liver disease worldwide. An improved understanding of the HCV replication cycle will enable development of novel and improved antiviral strategies. Here we have developed complementary fluorescent labeling and imaging approaches to investigate the localization, traffic and interactions of the HCV NS5A protein in living, virus-producing cells. These studies reveal new details as to the traffic, composition and biogenesis of NS5A foci and the nature of their association with putative sites of virus assembly. PMID:24429364

  6. Identification and characterization of two cleavage fragments from the Aquareovirus nonstructural protein NS80.

    PubMed

    Chen, Qingxiu; Zhang, Jie; Zhang, Fuxian; Guo, Hong; Fang, Qin

    2016-08-01

    Aquareovirus species vary with respect to pathogenicity, and the nonstructural protein NS80 of aquareoviruses has been implicated in the regulation of viral replication and assembly, which can form viral inclusion bodies (VIBs) and recruit viral proteins to its VIBs in infected cells. NS80 consists of 742 amino acids with a molecular weight of approximately 80 kDa. Interestingly, a short specific fragment of NS80 has also been detected in infected cells. In this study, an approximately 58-kDa product of NS80 was confirmed in various infected and transfected cells by immunoblotting analyses using α-NS80C. Mutational analysis and time course expression assays indicated that the accumulation of the 58-kDa fragment was related to time and infection dose, suggesting that the fragment is not a transient intermediate of protein degradation. Moreover, another smaller fragment with a molecular mass of approximately 22 kDa was observed in transfected and infected cells by immunoblotting with a specific anti-FLAG monoclonal antibody or α-NS80N, indicating that the 58- kDa polypeptide is derived from a specific cleavage site near the amino terminus of NS80. Additionally, different subcellular localization patterns were observed for the 22-kDa and 58-kDa fragments in an immunofluorescence analysis, implying that the two cleavage fragments of NS80 function differently in the viral life cycle. These results provide a basis for additional studies of the role of NS80 played in replication and particle assembly of the Aquareovirus. PMID:27279144

  7. Plasma Membrane Permeabilization by 60- and 600-ns Electric Pulses Is Determined by the Absorbed Dose

    PubMed Central

    Ibey, Bennett L.; Xiao, Shu; Schoenbach, Karl H.; Murphy, Michael R.; Pakhomov, Andrei G.

    2008-01-01

    We explored how the effect of plasma membrane permeabilization by nanosecond-duration electric pulses (nsEP) depends on the physical characteristics of exposure. The resting membrane resistance (Rm) and membrane potential (MP) were measured in cultured GH3 and CHO cells by conventional whole-cell patch-clamp technique. Intact cells were exposed to a single nsEP (60 or 600 ns duration, 0-22 kV/cm), followed by patch-clamp measurements after a 2-3 min delay. Consistent with earlier findings, nsEP caused long-lasting Rm decrease, accompanied by the loss of MP. The threshold for these effects was about 6 kV/cm for 60 ns pulses, and about 1 kV/cm for 600 ns pulses. Further analysis established that it was neither pulse duration nor the E-field amplitude per se, but the absorbed dose that determined the magnitude of the biological effect. In other words, exposure to nsEP at either pulse duration caused equal effects if the absorbed doses were equal. The threshold absorbed dose to produce plasma membrane effects in either GH3 or CHO cells at either pulse duration was found to be at or below 10 mJ/g. Despite being determined by the dose, the nsEP effect clearly is not thermal, as the maximum heating at the threshold dose is less than 0.01 °C. The use of the absorbed dose as a universal exposure metric may help to compare and quantify nsEP sensitivity of different cell types and of cells in different physiological conditions. The absorbed dose may also prove to be a more useful metric than the incident E-field in determining safety limits for high peak, lowaverage power EMF emissions. PMID:18839412

  8. Induction of Cell Death Mechanisms and Apoptosis by Nanosecond Pulsed Electric Fields (nsPEFs)

    PubMed Central

    Beebe, Stephen J.; Sain, Nova M.; Ren, Wei

    2013-01-01

    Pulse power technology using nanosecond pulsed electric fields (nsPEFs) offers a new stimulus to modulate cell functions or induce cell death for cancer cell ablation. New data and a literature review demonstrate fundamental and basic cellular mechanisms when nsPEFs interact with cellular targets. NsPEFs supra-electroporate cells creating large numbers of nanopores in all cell membranes. While nsPEFs have multiple cellular targets, these studies show that nsPEF-induced dissipation of ΔΨm closely parallels deterioration in cell viability. Increases in intracellular Ca2+ alone were not sufficient for cell death; however, cell death depended of the presence of Ca2+. When both events occur, cell death ensues. Further, direct evidence supports the hypothesis that pulse rise-fall times or high frequency components of nsPEFs are important for decreasing ΔΨm and cell viability. Evidence indicates in Jurkat cells that cytochrome c release from mitochondria is caspase-independent indicating an absence of extrinsic apoptosis and that cell death can be caspase-dependent and –independent. The Ca2+ dependence of nsPEF-induced dissipation of ΔΨm suggests that nanoporation of inner mitochondria membranes is less likely and effects on a Ca2+-dependent protein(s) or the membrane in which it is embedded are more likely a target for nsPEF-induced cell death. The mitochondria permeability transition pore (mPTP) complex is a likely candidate. Data demonstrate that nsPEFs can bypass cancer mutations that evade apoptosis through mechanisms at either the DISC or the apoptosome. PMID:24709649

  9. Fast Hepatitis C Virus RNA Elimination and NS5A Redistribution by NS5A Inhibitors Studied by a Multiplex Assay Approach

    PubMed Central

    Liu, Dandan; Ji, Juan; Ndongwe, Tanya P.; Michailidis, Eleftherios; Rice, Charles M.; Ralston, Robert

    2015-01-01

    While earlier therapeutic strategies for the treatment of hepatitis C virus (HCV) infection relied exclusively on interferon (IFN) and ribavirin (RBV), four direct-acting antiviral agents (DAAs) have now been approved, aiming for an interferon-free strategy with a short treatment duration and fewer side effects. To facilitate studies on the mechanism of action (MOA) and efficacy of DAAs, we established a multiplex assay approach, which employs flow cytometry, a Gaussia luciferase reporter system, Western blot analysis, reverse transcription-quantitative PCR (RT-qPCR), a limited dilution assay (50% tissue culture infectious dose [TCID50]), and an image profiling assay that follows the NS5A redistribution in response to drug treatment. We used this approach to compare the relative potency of various DAAs and the kinetics of their antiviral effects as a potential preclinical measure of their potential clinical utility. We evaluated the NS5A inhibitors ledipasvir (LDV) and daclatasvir (DCV), the NS3/4A inhibitor danoprevir (DNV), and the NS5B inhibitor sofosbuvir (SOF). In terms of kinetics, our data demonstrate that the NS5A inhibitor LDV, followed closely by DCV, has the fastest effect on suppression of viral proteins and RNA and on redistribution of NS5A. In terms of MOA, LDV has a more pronounced effect than DCV on the viral replication, assembly, and infectivity of released virus. Our approach can be used to facilitate the study of the biological processes involved in HCV replication and help identify optimal drug combinations. PMID:25845863

  10. Fast hepatitis C virus RNA elimination and NS5A redistribution by NS5A inhibitors studied by a multiplex assay approach.

    PubMed

    Liu, Dandan; Ji, Juan; Ndongwe, Tanya P; Michailidis, Eleftherios; Rice, Charles M; Ralston, Robert; Sarafianos, Stefan G

    2015-01-01

    While earlier therapeutic strategies for the treatment of hepatitis C virus (HCV) infection relied exclusively on interferon (IFN) and ribavirin (RBV), four direct-acting antiviral agents (DAAs) have now been approved, aiming for an interferon-free strategy with a short treatment duration and fewer side effects. To facilitate studies on the mechanism of action (MOA) and efficacy of DAAs, we established a multiplex assay approach, which employs flow cytometry, a Gaussia luciferase reporter system, Western blot analysis, reverse transcription-quantitative PCR (RT-qPCR), a limited dilution assay (50% tissue culture infectious dose [TCID50]), and an image profiling assay that follows the NS5A redistribution in response to drug treatment. We used this approach to compare the relative potency of various DAAs and the kinetics of their antiviral effects as a potential preclinical measure of their potential clinical utility. We evaluated the NS5A inhibitors ledipasvir (LDV) and daclatasvir (DCV), the NS3/4A inhibitor danoprevir (DNV), and the NS5B inhibitor sofosbuvir (SOF). In terms of kinetics, our data demonstrate that the NS5A inhibitor LDV, followed closely by DCV, has the fastest effect on suppression of viral proteins and RNA and on redistribution of NS5A. In terms of MOA, LDV has a more pronounced effect than DCV on the viral replication, assembly, and infectivity of released virus. Our approach can be used to facilitate the study of the biological processes involved in HCV replication and help identify optimal drug combinations. PMID:25845863

  11. Bluetongue Virus NS4 Protein Is an Interferon Antagonist and a Determinant of Virus Virulence

    PubMed Central

    Ratinier, Maxime; Shaw, Andrew E.; Barry, Gerald; Gu, Quan; Di Gialleonardo, Luigina; Janowicz, Anna; Varela, Mariana; Randall, Richard E.; Caporale, Marco

    2016-01-01

    ABSTRACT Bluetongue virus (BTV) is the causative agent of bluetongue, a major infectious disease of ruminants with serious consequences to both animal health and the economy. The clinical outcome of BTV infection is highly variable and dependent on a variety of factors related to both the virus and the host. In this study, we show that the BTV nonstructural protein NS4 favors viral replication in sheep, the animal species most affected by bluetongue. In addition, NS4 confers a replication advantage on the virus in interferon (IFN)-competent primary sheep endothelial cells and immortalized cell lines. We determined that in cells infected with an NS4 deletion mutant (BTV8ΔNS4), there is increased synthesis of type I IFN compared to cells infected with wild-type BTV-8. In addition, using RNA sequencing (RNA-seq), we show that NS4 modulates the host IFN response and downregulates mRNA levels of type I IFN and interferon-stimulated genes. Moreover, using reporter assays and protein synthesis assays, we show that NS4 downregulates the activities of a variety of promoters, such as the cytomegalovirus immediate-early promoter, the IFN-β promoter, and a promoter containing interferon-stimulated response elements (ISRE). We also show that the NS4 inhibitory activity on gene expression is related to its nucleolar localization. Furthermore, NS4 does not affect mRNA splicing or cellular translation. The data obtained in this study strongly suggest that BTV NS4 is an IFN antagonist and a key determinant of viral virulence. IMPORTANCE Bluetongue is one of the main infectious diseases of ruminants and is caused by bluetongue virus (BTV), an arthropod-borne virus transmitted from infected to susceptible animals by Culicoides biting midges. Bluetongue has a variable clinical outcome that can be related to both virus and host factors. It is therefore critical to understand the interplay between BTV and the host immune responses. In this study, we show that a nonstructural protein

  12. Flexibility of NS5 Methyltransferase-Polymerase Linker Region Is Essential for Dengue Virus Replication

    PubMed Central

    Zhao, Yongqian; Soh, Tingjin Sherryl; Chan, Kitti Wing Ki; Fung, Sarah Suet Yin; Swaminathan, Kunchithapadam; Lim, Siew Pheng; Shi, Pei-Yong; Huber, Thomas; Lescar, Julien

    2015-01-01

    We examined the function of the conserved Val/Ile residue within the dengue virus NS5 interdomain linker (residues 263 to 272) by site-directed mutagenesis. Gly substitution or Gly/Pro insertion after the conserved residue increased the linker flexibility and created slightly attenuated viruses. In contrast, Pro substitution abolished virus replication by imposing rigidity in the linker and restricting NS5's conformational plasticity. Our biochemical and reverse genetics experiments demonstrate that NS5 utilizes conformational regulation to achieve optimum viral replication. PMID:26269182

  13. Evidence for the existence of an Ns-type regulatory protein in Trypanosoma cruzi membranes.

    PubMed Central

    Eisenschlos, C D; Paladini, A A; Molina y Vedia, L; Torres, H N; Flawiá, M M

    1986-01-01

    The existence of a GTP-binding protein of the Ns type in Trypanosoma cruzi was explored. Epimastigote membranes were labelled by cholera toxin in the presence of [adenine-14C]NAD+. After SDS/polyacrylamide-gel electrophoresis of extracted membrane proteins, a single labelled polypeptide band of apparent Mr approx. 45,000 was detected. Epimastigote cells were treated with N-ethylmaleimide and electrofused to lymphoma S49 cells lacking the Ns protein. Evidence indicates that in such electrofusion-generated cell hybrids a heterologous adenylate cyclase system was reconstituted with the Ns protein provided by T. cruzi epimastigotes. Images Fig. 2. PMID:3099761

  14. A Neurodynamical Model of Brightness Induction in V1

    PubMed Central

    Penacchio, Olivier; Otazu, Xavier; Dempere-Marco, Laura

    2013-01-01

    Brightness induction is the modulation of the perceived intensity of an area by the luminance of surrounding areas. Recent neurophysiological evidence suggests that brightness information might be explicitly represented in V1, in contrast to the more common assumption that the striate cortex is an area mostly responsive to sensory information. Here we investigate possible neural mechanisms that offer a plausible explanation for such phenomenon. To this end, a neurodynamical model which is based on neurophysiological evidence and focuses on the part of V1 responsible for contextual influences is presented. The proposed computational model successfully accounts for well known psychophysical effects for static contexts and also for brightness induction in dynamic contexts defined by modulating the luminance of surrounding areas. This work suggests that intra-cortical interactions in V1 could, at least partially, explain brightness induction effects and reveals how a common general architecture may account for several different fundamental processes, such as visual saliency and brightness induction, which emerge early in the visual processing pathway. PMID:23717536

  15. PROFFIT: Analysis of X-ray surface-brightness profiles

    NASA Astrophysics Data System (ADS)

    Eckert, Dominique

    2016-08-01

    PROFFIT analyzes X-ray surface-brightness profiles for data from any X-ray instrument. It can extract surface-brightness profiles in circular or elliptical annuli, using constant or logarithmic bin size, from the image centroid, the surface-brightness peak, or any user-given center, and provides surface-brightness profiles in any circular or elliptical sectors. It offers background map support to extract background profiles, can excise areas using SAO DS9-compatible (ascl:0003.002) region files to exclude point sources, provides fitting with a number of built-in models, including the popular beta model, double beta, cusp beta, power law, and projected broken power law, uses chi-squared or C statistic, and can fit on the surface-brightness or counts data. It has a command-line interface similar to HEASOFT’s XSPEC (ascl:9910.005) package, provides interactive help with a description of all the commands, and results can be saved in FITS, ROOT or TXT format.

  16. Counting unstained, confluent cells by modified bright-field microscopy

    PubMed Central

    Drey, L. Louis; Graber, Michael C.; Bieschke, Jan

    2013-01-01

    We present a very simple procedure yielding high-contrast images of adherent, confluent cells such as human neuroblastoma (SH-EP) cells by ordinary bright-field microscopy. Cells are illuminated through a color filter and a pinhole aperture placed between the condenser and the cell culture surface. Refraction by each cell body generates a sharp, bright spot when the image is defocused. The technique allows robust, automatic cell counting from a single bright-field image in a wide range of focal positions; it does this via free, readily available image-analysis tools. Contrast may be enhanced by swelling cell bodies by brief incubation in PBS. The procedure was benchmarked against manual counting and automated counting of fluorescently labeled cell nuclei.. Counts from day-old and freshly seeded plates were compared in a range of densities, from sparse to densely overgrown. On average bright-field images produced the same counts as fluorescent images, with less than 5% error. This method will allow routine cell counting using a plain bright-field microscope, absent cell-line modification or cell staining. PMID:23834382

  17. Oxygen abundances in low surface-brightness galaxies

    NASA Technical Reports Server (NTRS)

    Roennback, Jari

    1993-01-01

    Recent theories predict that some protogalaxies, in low-density environments of the field, are contracting and interacting so slowly that global star formation can be delayed until today. These systems should be gas rich and have low surface-brightness. Blue compact galaxies (BCG's) and other compact HII region galaxies currently experiencing a burst of star formation are good candidates of truly young galaxies (in the sense that global star formation recently has been initiated). If they really are young, they ought to have a recent phase when their brightness was much lower than in the bursting phase. No claims of observations of such proto-BCG's exist. Observations of galaxies in their juvenile phases would undoubtedly be of great interest, e.g. the determination of the primordial helium abundance would improve. A proper place to search for young nearby galaxies could be among blue low surface-brightness galaxies (BLSBG's) in the local field. The study of low surface-brightness galaxies (LSBG's) as a group began relatively recently. They are galaxies with extraordinary properties both as individuals and as a group. A few years ago we started an optical study of a sample of BLSBG's selected from the ESO/Uppsala catalogue. Results of spectroscopic observations obtained on a subsample - 8 galaxies - of our selection are reported. The HII region oxygen chemical abundances and its relation to the blue absolute magnitude and surface-brightness is investigated.

  18. Bright and Not-So-Bright Prospects for Women in Physics in China-Beijing

    NASA Astrophysics Data System (ADS)

    Wu, Ling-An; Yang, Zhongqin; Ma, Wanyun

    2009-04-01

    Science in China-Beijing is enjoying a healthy increase in funding year by year, so the prospects for physicists are also bright. However, employment discrimination against women, formerly unthinkable, is becoming more and more explicit as the country evolves toward a market economy. Some recruitment notices bluntly state that only men will be considered, or impose restrictions upon potential female candidates. Female associate professors in many institutions are forced to retire at age 55, compared with 60 for men. This double-pinching discrimination against both younger and older women threatens to lead to a "pincer" effect, more serious than the "scissors" effect. Indeed, the ratio of senior-level women physicists in general has dropped significantly in recent years in China. Ironically, the number of female students applying for graduate studies is on the rise, as it is becoming increasingly difficult for them to compete with men in the job market with just an undergraduate degree. The Chinese Physical Society has made certain efforts to promote the image of women physicists, but it will take time and effort to reverse the trend.

  19. [The current state of research in bright light therapy].

    PubMed

    Bassa, Daniela; Canazei, Markus; Hinterhuber, Hartmann; Weiss, Elisabeth M

    2013-01-01

    The significance of light for the human organism and especially for the mental health is well-established for a long time. Therefore, the impact of light on mood and the use of bright light as a treatment-option for affective disorders have been studied extensively by scientists. Today bright light therapy is the treatment of choice for saisonal affective disorders. In the last years several clinical trials could demonstrate the therapeutic efficacy of bright light therapy for different neurological and psychiatric disorders such as sleep disorders, non-seasonal affective disorders or dementia. This article will give an overview about the neurobiological basis for light therapy and discuss different disorders responsive to light therapy. Finally a short overview about technical aspects of light therapy and new developments in light engineering will be presented.

  20. The effect of cirrus clouds on 118-GHz brightness temperatures

    NASA Technical Reports Server (NTRS)

    Weinman, J. A.

    1988-01-01

    A microwave radiative transfer model that describes the effect of scattering by cirrus clouds on the brightness temperatures that may be measured by a geostationary temperature-sounding radiometer has been developed. The model assumes that cirrus clouds are situated at an altitude where the temperature is about 230 K above completely absorbing land surfaces. It is shown that the brightness temperature at 118.75 + or - 3.9 GHz is depressed by 0.3-0.4 K per g per sq m of ice. The channels that operate at frequencies closer to 118.75 GHz are less affected by cirrus clouds. The brightness temperature reduction is most pronounced in warm temperate and tropical conditions.

  1. New low surface brightness dwarf galaxies detected around nearby spirals

    NASA Astrophysics Data System (ADS)

    Karachentsev, I. D.; Riepe, P.; Zilch, T.; Blauensteiner, M.; Elvov, M.; Hochleitner, P.; Hubl, B.; Kerschhuber, G.; Küppers, S.; Neyer, F.; Pölzl, R.; Remmel, P.; Schneider, O.; Sparenberg, R.; Trulson, U.; Willems, G.; Ziegler, H.

    2015-10-01

    We conduct a survey of low surface brightness (LSB) satellite galaxies around the Local Volume massive spirals using long exposures with small amateur telescopes. We identified 27 low and very low surface brightness objects around the galaxies NGC672, 891, 1156, 2683, 3344, 4258, 4618, 4631, and 5457 situated within 10 Mpc from us, and found nothing new around NGC2903, 3239, 4214, and 5585. Assuming that the dwarf candidates are the satellites of the neighboring luminous galaxies, their absolute magnitudes are in the range of -8.6 > M B > -13.3, their effective diameters are 0.4-4.7 kpc, and the average surface brightness is 26ṃ1/□″. The mean linear projected separation of the satellite candidates from the host galaxies is 73 kpc. Our spectroscopic observations of two LSB dwarfs with the Russian 6-meter telescope confirm their physical connection to the host galaxies NGC891 and NGC2683.

  2. A catalog of low surface brightness galaxies - List II

    NASA Technical Reports Server (NTRS)

    Schombert, James M.; Bothun, Gregory D.; Schneider, Stephen E.; Mcgaugh, Stacy S.

    1992-01-01

    A list of galaxies characterized by low surface brightness (LSB) is presented which facilitates the recognition of galaxies with brightnesses close to that of the sky. A total of 198 objects and 140 objects are listed in the primary and secondary catalogs respectively, and LSB galaxies are examined by means of H I redshift distributions. LSB disk galaxies are shown to have similar sizes and masses as the high-surface-brightness counterparts, and ellipticals and SOs are rarely encountered. Many LSB spirals have stellarlike nuclei, and most of the galaxies in the present catalog are late-type galaxies in the Sc, Sm, and Im classes. The LSB region of observational parameter space is shown to encompass a spectrum of types as full as that of the Hubble sequence. It is suggested that studies of LSB galaxies can provide important data regarding the formation and star-formation history of all galaxies.

  3. Bright Stuff on Ceres = Sulfates and Carbonates on CI Chondrites

    NASA Technical Reports Server (NTRS)

    Zolensky, Michael; Chan, Queenie H. S.; Gounelle, Matthieu; Fries, Marc

    2016-01-01

    Recent reports of the DAWN spacecraft's observations of the surface of Ceres indicate that there are bright areas, which can be explained by large amounts of the Mg sulfate hexahydrate (MgSO4•6(H2O)), although the identification appears tenuous. There are preliminary indications that water is being evolved from these bright areas, and some have inferred that these might be sites of contemporary hydro-volcanism. A heat source for such modern activity is not obvious, given the small size of Ceres, lack of any tidal forces from nearby giant planets, probable age and presumed bulk composition. We contend that observations of chondritic materials in the lab shed light on the nature of the bright spots on Ceres

  4. Dengue Virus NS Proteins Inhibit RIG-I/MAVS Signaling by Blocking TBK1/IRF3 Phosphorylation: Dengue Virus Serotype 1 NS4A Is a Unique Interferon-Regulating Virulence Determinant

    PubMed Central

    Dalrymple, Nadine A.; Cimica, Velasco

    2015-01-01

    ABSTRACT Dengue virus (DENV) replication is inhibited by the prior addition of type I interferon or by RIG-I agonists that elicit RIG-I/MAVS/TBK1/IRF3-dependent protective responses. DENV infection of primary human endothelial cells (ECs) results in a rapid increase in viral titer, which suggests that DENV inhibits replication-restrictive RIG-I/interferon beta (IFN-β) induction pathways within ECs. Our findings demonstrate that DENV serotype 4 (DENV4) nonstructural (NS) proteins NS2A and NS4B inhibited RIG-I-, MDA5-, MAVS-, and TBK1/IKKε-directed IFN-β transcription (>80%) but failed to inhibit IFN-β induction directed by STING or constitutively active IRF3-5D. Expression of NS2A and NS4B dose dependently inhibited the phosphorylation of TBK1 and IRF3, which suggests that they function at the level of TBK1 complex activation. NS2A and NS4B from DENV1/2/4, as well as the West Nile virus NS4B protein, commonly inhibited TBK1 phosphorylation and IFN-β induction. A comparative analysis of NS4A proteins across DENVs demonstrated that DENV1, but not DENV2 or DENV4, NS4A proteins uniquely inhibited TBK1. These findings indicate that DENVs contain conserved (NS2A/NS4B) and DENV1-specific (NS4A) mechanisms for inhibiting RIG-I/TBK1-directed IFN responses. Collectively, our results define DENV NS proteins that restrict IRF3 and IFN responses and thereby facilitate DENV replication and virulence. Unique DENV1-specific NS4A regulation of IFN induction has the potential to be a virulence determinant that contributes to the increased severity of DENV1 infections and the immunodominance of DENV1 responses during tetravalent DENV1-4 vaccination. PMID:25968648

  5. [Characterization of a panel of monoclonal antibodies to hepatitis C NS3 recombinant protein ].

    PubMed

    Abdulmedzhidova, A G; Masalova, O V; Atanadze, S N; Ulanova, T I; Burkov, A N; Khudiakov, Iu E; Fields, H; Kushch, A A

    2002-01-01

    Recombinant protein rNS3 imitating helicase region (1356-1459 amino acid residues) of hepatitis C virus (HCV) was expressed in E. coli cells and used for BALB/c mice immunization. Seven hybrydoma clones producing monoclonal antibodies (MAbs) to rHS3 were obtained. All MAbs reacted in ELISA with NS3 protein from Murex anti-HCV Version III and in immunoblotting from RIBA 3. These MAbs detect 5 individual epitopes, 4 of which were conformational and 1 discontinuous. All MAbs could compete for rNS3 binding with serum antibodies from patients with chronic hepatitis C, which suggests that these MAbs can recognize the natural HCV NS3 protein.

  6. The Influenza NS1 Protein: What Do We Know in Equine Influenza Virus Pathogenesis?

    PubMed

    Barba, Marta; Daly, Janet M

    2016-08-31

    Equine influenza virus remains a serious health and potential economic problem throughout most parts of the world, despite intensive vaccination programs in some horse populations. The influenza non-structural protein 1 (NS1) has multiple functions involved in the regulation of several cellular and viral processes during influenza infection. We review the strategies that NS1 uses to facilitate virus replication and inhibit antiviral responses in the host, including sequestering of double-stranded RNA, direct modulation of protein kinase R activity and inhibition of transcription and translation of host antiviral response genes such as type I interferon. Details are provided regarding what it is known about NS1 in equine influenza, especially concerning C-terminal truncation. Further research is needed to determine the role of NS1 in equine influenza infection, which will help to understand the pathophysiology of complicated cases related to cytokine imbalance and secondary bacterial infection, and to investigate new therapeutic and vaccination strategies.

  7. The Influenza NS1 Protein: What Do We Know in Equine Influenza Virus Pathogenesis?

    PubMed Central

    Barba, Marta; Daly, Janet M.

    2016-01-01

    Equine influenza virus remains a serious health and potential economic problem throughout most parts of the world, despite intensive vaccination programs in some horse populations. The influenza non-structural protein 1 (NS1) has multiple functions involved in the regulation of several cellular and viral processes during influenza infection. We review the strategies that NS1 uses to facilitate virus replication and inhibit antiviral responses in the host, including sequestering of double-stranded RNA, direct modulation of protein kinase R activity and inhibition of transcription and translation of host antiviral response genes such as type I interferon. Details are provided regarding what it is known about NS1 in equine influenza, especially concerning C-terminal truncation. Further research is needed to determine the role of NS1 in equine influenza infection, which will help to understand the pathophysiology of complicated cases related to cytokine imbalance and secondary bacterial infection, and to investigate new therapeutic and vaccination strategies. PMID:27589809

  8. Influenza virus NS1 protein interacts with viral transcription-replication complexes in vivo.

    PubMed

    Marión, R M; Zürcher, T; de la Luna, S; Ortín, J

    1997-10-01

    The interaction of influenza virus NS1 protein with other viral products in the infected cell was analysed by co-immunoprecipitation studies. The three subunits of the polymerase and the nucleoprotein, but not M1 protein, were co-immunoprecipitated by NS1-specific serum but not when control serum was used. Such co-immunoprecipitation was not sensitive to RNase treatment of the immunoprecipitates. Co-immunoprecipitation was also obtained when the viral transcription-replication system was reconstituted in vivo by transfection of cDNAs and model vRNA template into vaccinia virus-T7-infected cells. Analysis of the RNA pulled-down in the NS1-specific precipitates indicated the presence of both vRNA and mRNA. These results are discussed in the context of the phenotype of virus temperature-sensitive mutants affected in the NS1 gene.

  9. Study of anti-dengue NS1 antibody by western blot.

    PubMed

    Kuno, G; Vorndam, A V; Gubler, D J; Gómez, I

    1990-10-01

    The presence of anti-nonstructural protein (NS1) antibody in natural dengue infections has been suspected to be associated with development of dengue haemorrhagic fever (DHF). The Western blot technique was used to study the dynamics of the immune response to NS1 and to determine the frequency of anti-NS1 antibody among confirmed dengue patients in Indonesia, where DHF is common, and in Puerto Rico, where DHF is less frequently observed. Anti-NS1 antibody was rarely found in those with primary infections in either group. The antibody occurred at a significantly higher frequency in acute-phase serum samples from secondary infections in Indonesia than in those from Puerto Rico. No difference was observed, however, in Indonesian patients with secondary infection who had dengue fever or DHF.

  10. The NS3 protein of rice hoja blanca virus suppresses RNA silencing in mammalian cells.

    PubMed

    Schnettler, Esther; Hemmes, Hans; Goldbach, Rob; Prins, Marcel

    2008-01-01

    The NS3 protein of the tenuivirus rice hoja blanca virus (RHBV) has previously been shown to represent the viral RNA interference (RNAi) suppressor and is active in both plant and insect cells by binding short interfering RNAs (siRNAs) in vitro. Using a firefly luciferase-based silencing assay it is described here that NS3 is also active in mammalian cells. This activity is independent of the inducer molecule used. Using either synthetic siRNAs or a short hairpin RNA construct, NS3 was able to significantly suppress the RNAi-mediated silencing of luciferase expression in both monkey (Vero) and human (HEK293) cells. These results support the proposed mode of action of NS3 to act by sequestering siRNAs, the key molecules of the RNAi pathway conserved in all eukaryotes. The possible applications of this protein in modulating RNAi and investigating the proposed antiviral RNAi response in mammalian cell systems are discussed. PMID:18089758

  11. Detection of dengue NS1 antigen using long-range surface plasmon waveguides.

    PubMed

    Wong, Wei Ru; Sekaran, Shamala Devi; Adikan, Faisal Rafiq Mahamd; Berini, Pierre

    2016-04-15

    The non-structural 1 (NS1) protein of the dengue virus circulates in infected patients' blood samples and can be used for early diagnosis of dengue infection. In this paper, we present the detection of naturally-occurring dengue NS1 antigen in infected patient blood plasma using straight long-range surface plasmon waveguides. Three commercially-available anti-NS1 monoclonal antibodies were used for recognition and their performance was compared and discussed. A similar figure of merit to the one used in conventional dengue NS1 capture using an enzyme-linked immunosorbent assay (ELISA) was applied to our results. In general, the positive patient samples can be clearly differentiated from the negative ones and the results agree with those obtained using ELISA. The largest signal-to-noise ratio observed during the experiments was 356 and the best detection limit observed is estimated as 5.73 pg/mm(2).

  12. Influenza A Virus Attenuation by Codon Deoptimization of the NS Gene for Vaccine Development

    PubMed Central

    Nogales, Aitor; Baker, Steven F.; Ortiz-Riaño, Emilio; Dewhurst, Stephen; Topham, David J.

    2014-01-01

    ABSTRACT Influenza viral infection represents a serious public health problem that causes contagious respiratory disease, which is most effectively prevented through vaccination to reduce transmission and future infection. The nonstructural (NS) gene of influenza A virus encodes an mRNA transcript that is alternatively spliced to express two viral proteins, the nonstructural protein 1 (NS1) and the nuclear export protein (NEP). The importance of the NS gene of influenza A virus for viral replication and virulence has been well described and represents an attractive target to generate live attenuated influenza viruses with vaccine potential. Considering that most amino acids can be synthesized from several synonymous codons, this study employed the use of misrepresented mammalian codons (codon deoptimization) for the de novo synthesis of a viral NS RNA segment based on influenza A/Puerto Rico/8/1934 (H1N1) (PR8) virus. We generated three different recombinant influenza PR8 viruses containing codon-deoptimized synonymous mutations in coding regions comprising the entire NS gene or the mRNA corresponding to the individual viral protein NS1 or NEP, without modifying the respective splicing and packaging signals of the viral segment. The fitness of these synthetic viruses was attenuated in vivo, while they retained immunogenicity, conferring both homologous and heterologous protection against influenza A virus challenges. These results indicate that influenza viruses can be effectively attenuated by synonymous codon deoptimization of the NS gene and open the possibility of their use as a safe vaccine to prevent infections with these important human pathogens. IMPORTANCE Vaccination serves as the best therapeutic option to protect humans against influenza viral infections. However, the efficacy of current influenza vaccines is suboptimal, and novel approaches are necessary for the prevention of disease cause by this important human respiratory pathogen. The nonstructural

  13. Brightness discrimination in the South African fur seal (Arctocephalus pusillus).

    PubMed

    Scholtyssek, C; Dehnhardt, G

    2013-05-24

    Underwater, the contrast between object and background is much larger reduced with increasing distance between object and observer than in air. For marine predators, such as pinnipeds, it would therefore be advantageous to possess a high sensitivity for brightness differences, since this would increase the distance at which prey can be detected visually. Few studies have examined the brightness discrimination thresholds of pinnipeds. Two studies with phocid seals have confirmed low brightness discrimination thresholds in pinnipeds whereas the threshold obtained for the South African fur seal seems to be twice as high as that of the phocids. However, the experiments with the South African fur seal have been conducted under inadequate conditions which likely resulted in an underestimation of the brightness discrimination ability of this species. The study at hand reinvestigated the brightness discrimination threshold of the South African fur seal under well controlled conditions. In a two alternative forced choice task, one fur seal was trained to indicate the position of the brighter of two gray discs presented on a black background on a monitor. The thresholds were determined for 11 standard intensities each tested against 8 lower comparison intensities. It was found that the fur seal was able to perceive brightness differences of 8-10%, which is better than the phocid species tested so far. For low standard intensities, however, the threshold increased which could to be due to a relative slow dark adaptation rate of the fur seal. The results are discussed in terms of the relevance of visual information for pinnipeds during foraging dives and are directly compared to the results obtained for the harbor seal which has been tested under the same conditions as the fur seal in a previous study.

  14. The possible origin of facular brightness in the solar atmosphere

    NASA Astrophysics Data System (ADS)

    Kostik, R.; Khomenko, E.

    2016-05-01

    This paper studies the dependence of the Ca ii H line core brightness on the strength and inclination of the photospheric magnetic field, and on the parameters of convective and wave motions in a facular region at the center of the solar disc. We use three simultaneous data sets that were obtained at the German Vacuum Tower Telescope (Observatorio del Teide, Tenerife): (1) spectra of Ba ii 4554 Å line, registered with the instrument TESOS to measure the variations of intensity and velocity through the photosphere up to the temperature minimum; (2) spectropolarimetric data in Fe i 1.56 μm lines (registered with the instrument TIP II) to measure photospheric magnetic fields; (3) filtergrams in Ca ii H that give information about brightness fluctuations in the chromosphere. The results show that the Ca ii H brightness in the facula strongly depends on the power of waves with periods in the 5-min range, which propagate upwards, and also on the phase shift between velocity oscillations at the bottom photosphere and around the temperature minimum height that is measured from Ba ii line. The Ca ii H brightness is maximum at locations where the phase shift between temperature and velocity oscillations lies within 0°-100°. There is an indirect influence of convective motions on the Ca ii H brightness. The higher the amplitude of convective velocities is and the greater the height is where they change their direction of motion, the brighter the facula. In summary, our results lead to conclusions that facular regions appear bright not only because of the Wilson depression in magnetic structures, but also owing to real heating.

  15. Bright and dark excitons in semiconductor carbon nanotubes

    SciTech Connect

    Tretiak, Sergei

    2008-01-01

    We report electronic structure calculations of finite-length semiconducting carbon nanotubes using the time dependent density functional theory (TD-DFT) and the time dependent Hartree Fock (TD-HF) approach coupled with semiempirical AM1 and ZINDO Hamiltonians. We specifically focus on the energy splitting, relative ordering, and localization properties of the optically active (bright) and optically forbidden (dark) states from the lowest excitonic band of the nanotubes. These excitonic states are very important in competing radiative and non-radiative processes in these systems. Our analysis of excitonic transition density matrices demonstrates that pure DFT functionals overdelocalize excitons making an electron-hole pair unbound; consequently, excitonic features are not presented in this method. In contrast, the pure HF and A111 calculations overbind excitons inaccurately predicting the lowest energy state as a bright exciton. Changing AM1 with ZINDO Hamiltonian in TD-HF calculations, predicts the bright exciton as the second state after the dark one. However, in contrast to AM1 calculations, the diameter dependence of the excitation energies obtained by ZINDO does not follow the experimental trends. Finally, the TD-DFT approach incorporating hybrid functions with a moderate portion of the long-range HF exchange, such as B3LYP, has the most generality and predictive capacity providing a sufficiently accurate description of excitonic structure in finite-size nanotubes. These methods characterize four important lower exciton bands. The lowest state is dark, the upper band is bright, and the two other dark and nearly degenerate excitons lie in-between. Although the calculated energy splittings between the lowest dark and the bright excitons are relatively large ({approx}0.1 eV), the dense excitonic manifold below the bright exciton allows for fast non-radiative relaxation leasing to the fast population of the lowest dark exciton. This rationalizes the low

  16. Effects of bright light treatment on psychomotor speed in athletes

    PubMed Central

    Tulppo, Mikko P.; Jurvelin, Heidi; Roivainen, Eka; Nissilä, Juuso; Hautala, Arto J.; Kiviniemi, Antti M.; Kiviniemi, Vesa J.; Takala, Timo

    2014-01-01

    Purpose: A recent study suggests that transcranial brain targeted light treatment via ear canals may have physiological effects on brain function studied by functional magnetic resonance imaging (fMRI) techniques in humans. We tested the hypothesis that bright light treatment could improve psychomotor speed in professional ice hockey players. Methods: Psychomotor speed tests with audio and visual warning signals were administered to a Finnish National Ice Hockey League team before and after 24 days of transcranial bright light or sham treatment. The treatments were given during seasonal darkness in the Oulu region (latitude 65 degrees north) when the strain on the players was also very high (10 matches during 24 days). A daily 12-min dose of bright light or sham (n = 11 for both) treatment was given every morning between 8 and 12 am at home with a transcranial bright light device. Mean reaction time and motor time were analyzed separately for both psychomotor tests. Analysis of variance for repeated measures adjusted for age was performed. Results: Time × group interaction for motor time with a visual warning signal was p = 0.024 after adjustment for age. In Bonferroni post-hoc analysis, motor time with a visual warning signal decreased in the bright light treatment group from 127 ± 43 to 94 ± 26 ms (p = 0.024) but did not change significantly in the sham group 121 ± 23 vs. 110 ± 32 ms (p = 0.308). Reaction time with a visual signal did not change in either group. Reaction or motor time with an audio warning signal did not change in either the treatment or sham group. Conclusion: Psychomotor speed, particularly motor time with a visual warning signal, improves after transcranial bright light treatment in professional ice-hockey players during the competition season in the dark time of the year. PMID:24860513

  17. Extended surface for membrane association in Zika virus NS1 structure.

    PubMed

    Brown, W Clay; Akey, David L; Konwerski, Jamie R; Tarrasch, Jeffrey T; Skiniotis, Georgios; Kuhn, Richard J; Smith, Janet L

    2016-09-01

    The Zika virus, which has been implicated in an increase in neonatal microcephaly and Guillain-Barré syndrome, has spread rapidly through tropical regions of the world. The virulence protein NS1 functions in genome replication and host immune-system modulation. Here, we report the crystal structure of full-length Zika virus NS1, revealing an elongated hydrophobic surface for membrane association and a polar surface that varies substantially among flaviviruses. PMID:27455458

  18. Ebselen Inhibits Hepatitis C Virus NS3 Helicase Binding to Nucleic Acid and Prevents Viral Replication

    PubMed Central

    2015-01-01

    The hepatitis C virus (HCV) nonstructural protein 3 (NS3) is both a protease, which cleaves viral and host proteins, and a helicase that separates nucleic acid strands, using ATP hydrolysis to fuel the reaction. Many antiviral drugs, and compounds in clinical trials, target the NS3 protease, but few helicase inhibitors that function as antivirals have been reported. This study focuses on the analysis of the mechanism by which ebselen (2-phenyl-1,2-benzisoselenazol-3-one), a compound previously shown to be a HCV antiviral agent, inhibits the NS3 helicase. Ebselen inhibited the abilities of NS3 to unwind nucleic acids, to bind nucleic acids, and to hydrolyze ATP, and about 1 μM ebselen was sufficient to inhibit each of these activities by 50%. However, ebselen had no effect on the activity of the NS3 protease, even at 100 times higher ebselen concentrations. At concentrations below 10 μM, the ability of ebselen to inhibit HCV helicase was reversible, but prolonged incubation of HCV helicase with higher ebselen concentrations led to irreversible inhibition and the formation of covalent adducts between ebselen and all 14 cysteines present in HCV helicase. Ebselen analogues with sulfur replacing the selenium were just as potent HCV helicase inhibitors as ebselen, but the length of the linker between the phenyl and benzisoselenazol rings was critical. Modifications of the phenyl ring also affected compound potency over 30-fold, and ebselen was a far more potent helicase inhibitor than other, structurally unrelated, thiol-modifying agents. Ebselen analogues were also more effective antiviral agents, and they were less toxic to hepatocytes than ebselen. Although the above structure–activity relationship studies suggest that ebselen targets a specific site on NS3, we were unable to confirm binding to either the NS3 ATP binding site or nucleic acid binding cleft by examining the effects of ebselen on NS3 proteins lacking key cysteines. PMID:25126694

  19. Zika virus NS1 structure reveals diversity of electrostatic surfaces among flaviviruses.

    PubMed

    Song, Hao; Qi, Jianxun; Haywood, Joel; Shi, Yi; Gao, George F

    2016-05-01

    The association of Zika virus (ZIKV) infections with microcephaly has resulted in an ongoing public-health emergency. Here we report the crystal structure of a C-terminal fragment of ZIKV nonstructural protein 1 (NS1), a major host-interaction molecule that functions in flaviviral replication, pathogenesis and immune evasion. Comparison with West Nile and dengue virus NS1 structures reveals conserved features but diverse electrostatic characteristics at host-interaction interfaces, thus possibly implying different modes of flavivirus pathogenesis.

  20. Extended surface for membrane association in Zika virus NS1 structure.

    PubMed

    Brown, W Clay; Akey, David L; Konwerski, Jamie R; Tarrasch, Jeffrey T; Skiniotis, Georgios; Kuhn, Richard J; Smith, Janet L

    2016-09-01

    The Zika virus, which has been implicated in an increase in neonatal microcephaly and Guillain-Barré syndrome, has spread rapidly through tropical regions of the world. The virulence protein NS1 functions in genome replication and host immune-system modulation. Here, we report the crystal structure of full-length Zika virus NS1, revealing an elongated hydrophobic surface for membrane association and a polar surface that varies substantially among flaviviruses.

  1. New binding site conformations of the dengue virus NS3 protease accessed by molecular dynamics simulation.

    PubMed

    de Almeida, Hugo; Bastos, Izabela M D; Ribeiro, Bergmann M; Maigret, Bernard; Santana, Jaime M

    2013-01-01

    Dengue fever is caused by four distinct serotypes of the dengue virus (DENV1-4), and is estimated to affect over 500 million people every year. Presently, there are no vaccines or antiviral treatments for this disease. Among the possible targets to fight dengue fever is the viral NS3 protease (NS3PRO), which is in part responsible for viral processing and replication. It is now widely recognized that virtual screening campaigns should consider the flexibility of target protein by using multiple active conformational states. The flexibility of the DENV NS3PRO could explain the relatively low success of previous virtual screening studies. In this first work, we explore the DENV NS3PRO conformational states obtained from molecular dynamics (MD) simulations to take into account protease flexibility during the virtual screening/docking process. To do so, we built a full NS3PRO model by multiple template homology modeling. The model comprised the NS2B cofactor (essential to the NS3PRO activation), a glycine flexible link and the proteolytic domain. MD simulations had the purpose to sample, as closely as possible, the ligand binding site conformational landscape prior to inhibitor binding. The obtained conformational MD sample was clustered into four families that, together with principal component analysis of the trajectory, demonstrated protein flexibility. These results allowed the description of multiple binding modes for the Bz-Nle-Lys-Arg-Arg-H inhibitor, as verified by binding plots and pair interaction analysis. This study allowed us to tackle protein flexibility in our virtual screening campaign against the dengue virus NS3 protease.

  2. Networks of Host Factors that Interact with NS1 Protein of Influenza A Virus

    PubMed Central

    Thulasi Raman, Sathya N.; Zhou, Yan

    2016-01-01

    Pigs are an important host of influenza A viruses due to their ability to generate reassortant viruses with pandemic potential. NS1 protein of influenza A viruses is a key virulence factor and a major antagonist of innate immune responses. It is also involved in enhancing viral mRNA translation and regulation of virus replication. Being a protein with pleiotropic functions, NS1 has a variety of cellular interaction partners. Hence, studies on swine influenza viruses (SIV) and identification of swine influenza NS1-interacting host proteins is of great interest. Here, we constructed a recombinant SIV carrying a Strep-tag in the NS1 protein and infected primary swine respiratory epithelial cells (SRECs) with this virus. The Strep-tag sequence in the NS1 protein enabled us to purify intact, the NS1 protein and its interacting protein complex specifically. We identified cellular proteins present in the purified complex by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and generated a dataset of these proteins. 445 proteins were identified by LC-MS/MS and among them 192 proteins were selected by setting up a threshold based on MS parameters. The selected proteins were analyzed by bioinformatics and were categorized as belonging to different functional groups including translation, RNA processing, cytoskeleton, innate immunity, and apoptosis. Protein interaction networks were derived using these data and the NS1 interactions with some of the specific host factors were verified by immunoprecipitation. The novel proteins and the networks revealed in our study will be the potential candidates for targeted study of the molecular interaction of NS1 with host proteins, which will provide insights into the identification of new therapeutic targets to control influenza infection and disease pathogenesis. PMID:27199973

  3. Novel influenza virus NS1 antagonists block replication and restore innate immune function.

    PubMed

    Basu, Dipanwita; Walkiewicz, Marcin P; Frieman, Matthew; Baric, Ralph S; Auble, David T; Engel, Daniel A

    2009-02-01

    The innate immune system guards against virus infection through a variety of mechanisms including mobilization of the host interferon system, which attacks viral products mainly at a posttranscriptional level. The influenza virus NS1 protein is a multifunctional facilitator of virus replication, one of whose actions is to antagonize the interferon response. Since NS1 is required for efficient virus replication, it was reasoned that chemical inhibitors of this protein could be used to further understand virus-host interactions and also serve as potential new antiviral agents. A yeast-based assay was developed to identify compounds that phenotypically suppress NS1 function. Several such compounds exhibited significant activity specifically against influenza A virus in cell culture but had no effect on the replication of another RNA virus, respiratory syncytial virus. Interestingly, cells lacking an interferon response were drug resistant, suggesting that the compounds block interactions between NS1 and the interferon system. Accordingly, the compounds reversed the inhibition of beta interferon mRNA induction during infection, which is known to be caused by NS1. In addition, the compounds blocked the ability of NS1 protein to inhibit double-stranded RNA-dependent activation of a transfected beta interferon promoter construct. The effects of the compounds were specific to NS1, because they had no effect on the ability of the severe acute respiratory syndrome coronavirus papainlike protease protein to block beta interferon promoter activation. These data demonstrate that the function of NS1 can be modulated by chemical inhibitors and that such inhibitors will be useful as probes of biological function and as starting points for clinical drug development.

  4. Hepatitis C Virus NS5A Inhibits Mixed Lineage Kinase 3 to Block Apoptosis*

    PubMed Central

    Amako, Yutaka; Igloi, Zsofia; Mankouri, Jamel; Kazlauskas, Arunas; Saksela, Kalle; Dallas, Mark; Peers, Chris; Harris, Mark

    2013-01-01

    Hepatitis C virus (HCV) infection results in the activation of numerous stress responses including oxidative stress, with the potential to induce an apoptotic state. Previously we have shown that HCV attenuates the stress-induced, p38MAPK-mediated up-regulation of the K+ channel Kv2.1, to maintain the survival of infected cells in the face of cellular stress. We demonstrated that this effect was mediated by HCV non-structural 5A (NS5A) protein, which impaired p38MAPK activity through a polyproline motif-dependent interaction, resulting in reduction of phosphorylation activation of Kv2.1. In this study, we investigated the host cell proteins targeted by NS5A to mediate Kv2.1 inhibition. We screened a phage-display library expressing the entire complement of human SH3 domains for novel NS5A-host cell interactions. This analysis identified mixed lineage kinase 3 (MLK3) as a putative NS5A interacting partner. MLK3 is a serine/threonine protein kinase that is a member of the MAPK kinase kinase (MAP3K) family and activates p38MAPK. An NS5A-MLK3 interaction was confirmed by co-immunoprecipitation and Western blot analysis. We further demonstrate a novel role of MLK3 in the modulation of Kv2.1 activity, whereby MLK3 overexpression leads to the up-regulation of channel activity. Accordingly, coexpression of NS5A suppressed this stimulation. Additionally we demonstrate that overexpression of MLK3 induced apoptosis, which was also counteracted by NS5A. We conclude that NS5A targets MLK3 with multiple downstream consequences for both apoptosis and K+ homeostasis. PMID:23857585

  5. Laser ion source for high brightness heavy ion beam

    NASA Astrophysics Data System (ADS)

    Okamura, M.

    2016-09-01

    A laser ion source is known as a high current high charge state heavy ion source. However we place great emphasis on the capability to realize a high brightness ion source. A laser ion source has a pinpoint small volume where materials are ionized and can achieve quite uniform low temperature ion beam. Those features may enable us to realize very small emittance beams. In 2014, a low charge state high brightness laser ion source was successfully commissioned in Brookhaven National Laboratory. Now most of all the solid based heavy ions are being provided from the laser ion source for regular operation.

  6. Dark dyes-bright complexes: fluorogenic protein labeling.

    PubMed

    Bruchez, Marcel P

    2015-08-01

    Complexes formed between organic dyes and genetically encoded proteins combine the advantages of stable and tunable fluorescent molecules and targetable, biologically integrated labels. To overcome the challenges imposed by labeling with bright fluorescent dyes, a number of approaches now exploit chemical or environmental changes to control the properties of a bound dye, converting dyes from a weakly fluorescent state to a bright, easily detectable complex. Optimized, such approaches avoid the need for removal of unbound dyes, facilitate rapid and simple assays in cultured cells and enable hybrid labeling to function more robustly in living model organisms.

  7. The brightness variations of Comet Halley at large heliocentric distances

    NASA Technical Reports Server (NTRS)

    Flammer, K. R.; Jackson, B.; Houpis, H. L. F.; Mendis, D. A.

    1986-01-01

    The reasons for the intrinsic brightness variations of up to 500 percent on time scales as short as a few hours detected by Sekanina (1984) in Comet Halley between October 1982 and February 1984 are discussed. It is shown that solar wind-modulated electrostatic dust blowoff from the night side of the comet is consistent with the observed brightness variations. The variations coincide with the encounter of high-speed streams with the comet. The stream's propagation time to the comet and the sun's rotation during this transit were used to locate the stream origin on the coronal surface, and the results are shown.

  8. Implications of Titan's north-south brightness asymmetry

    NASA Technical Reports Server (NTRS)

    Sromovsky, L. A.; Suomi, V. E.; Krauss, R. J.; Limaye, S. S.; Revercomb, H. E.; Pollack, J. B.; Owen, T.; Sagan, C.

    1981-01-01

    Voyager 1 images of Titan, when normalized to remove limb darkening, reveal an axially symmetric brightness pattern with significant north-south asymmetry. This interhemispheric contrast seems to be a response to seasonal solar heating variations resulting from Titan's inclined spin axis. The contrast significantly lags the solar forcing, indicating that its production involves the atmosphere well below the unit optical depth level. The contrast has a significant effect on Titan's disk-integrated brightness as seen from earth, and probably accounts for most of the observed long term variation, with solar UV variations accounting for the remainder.

  9. DUST-SCATTERED ULTRAVIOLET HALOS AROUND BRIGHT STARS

    SciTech Connect

    Murthy, Jayant; Henry, Richard Conn

    2011-06-10

    We have discovered ultraviolet (UV) halos extending as far as 5 deg. around four (of six) bright UV stars using data from the Galaxy Evolution Explorer satellite. These halos are due to scattering of the starlight from nearby thin, foreground dust clouds. We have placed limits of 0.58 {+-} 0.12 and 0.72 {+-} 0.06 on the phase function asymmetry factor (g) in the FUV (1521 A) and NUV (2320 A) bands, respectively. We suggest that these halos are a common feature around bright stars and may be used to explore the scattering function of interstellar grains at small angles.

  10. High-brightness displays in integrated weapon sight systems

    NASA Astrophysics Data System (ADS)

    Edwards, Tim; Hogan, Tim

    2014-06-01

    In the past several years Kopin has demonstrated the ability to provide ultra-high brightness, low power display solutions in VGA, SVGA, SXGA and 2k x 2k display formats. This paper will review various approaches for integrating high brightness overlay displays with existing direct view rifle sights and augmenting their precision aiming and targeting capability. Examples of overlay display systems solutions will be presented and discussed. This paper will review significant capability enhancements that are possible when augmenting the real-world as seen through a rifle sight with other soldier system equipment including laser range finders, ballistic computers and sensor systems.

  11. A bright PPKTP waveguide source of polarization entangled photons

    SciTech Connect

    Fanto, Michael; Tison, Christoper C.; Holwand, Gregory A; Preble, Dr. Stefan F; Alsing, Paul; Smith IV, Amos M

    2015-01-01

    The need for bright efficient sources of entangled photons has been a subject of tremendous research over the last decade. Researchers have been working to increase the brightness and purity to help overcome the spontaneous nature of the sources. Periodic poling has been implemented to allow for the use of crystals that would not normally satisfy the phase matching conditions. Utilizing periodic poling and single mode waveguide confinement of the pump field has yielded extremely large effective nonlinearities in sources easily producing millions of photon pairs. Here we will demonstrate these large nonlinearity effects in a periodically poled potassium titanyl phosphate (PPKTP) waveguide as well as characterizing the source purity.

  12. Are solar brightness variations faculae- or spot-dominated?

    NASA Astrophysics Data System (ADS)

    Shapiro, A. I.; Solanki, S. K.; Krivova, N. A.; Yeo, K. L.; Schmutz, W. K.

    2016-05-01

    Context. Regular spaceborne measurements have revealed that solar brightness varies on multiple timescales, variations on timescales greater than a day being attributed to a surface magnetic field. Independently, ground-based and spaceborne measurements suggest that Sun-like stars show a similar, but significantly broader pattern of photometric variability. Aims: To understand whether the broader pattern of stellar variations is consistent with the solar paradigm, we assess relative contributions of faculae and spots to solar magnetically-driven brightness variability. We investigate how the solar brightness variability and its facular and spot contributions depend on the wavelength, timescale of variability, and position of the observer relative to the ecliptic plane. Methods: We performed calculations with the SATIRE model, which returns solar brightness with daily cadence from solar disc area coverages of various magnetic features. We took coverages as seen by an Earth-based observer from full-disc SoHO/MDI and SDO/HMI data and projected them to mimic out-of-ecliptic viewing by an appropriate transformation. Results: Moving the observer away from the ecliptic plane increases the amplitude of 11-year variability as it would be seen in Strömgren (b + y)/2 photometry, but decreases the amplitude of the rotational brightness variations as it would appear in Kepler and CoRoT passbands. The spot and facular contributions to the 11-year solar variability in the Strömgren (b + y)/2 photometry almost fully compensate each other so that the Sun appears anomalously quiet with respect to its stellar cohort. Such a compensation does not occur on the rotational timescale. Conclusions: The rotational solar brightness variability as it would appear in the Kepler and CoRoT passbands from the ecliptic plane is spot-dominated, but the relative contribution of faculae increases for out-of-ecliptic viewing so that the apparent brightness variations are faculae-dominated for

  13. An updated evolutionary study of Flaviviridae NS3 helicase and NS5 RNA-dependent RNA polymerase reveals novel invariable motifs as potential pharmacological targets.

    PubMed

    Papageorgiou, Louis; Loukatou, Styliani; Sofia, Kossida; Maroulis, Dimitrios; Vlachakis, Dimitrios

    2016-06-21

    The rate of Flaviviridae family virus infections worldwide has increased dramatically in the last few years. In addition, infections caused by arthropod vector viruses including Hepatitis C, West Nile, Dengue fever, Yellow fever and Japanese encephalitis are emerging throughout the world. Based on a recent taxon update, the Flaviviridae family comprises four main genera; Flavivirus, Hepacivirus, Pestivirus and a recent genus Pegivirus. Although the new scientific classification plays a key role in providing useful information about the relationships between viruses, many new documented viruses remain unclassified. Furthermore, based on the different results of several studies the classification is unclear. In an effort to provide more insights into the classification of viruses, a holistic evolutionary study of the two viral enzymes NS3 helicase and NS5 RNA-dependent RNA polymerase (RdRp) has been conducted in this study. These two viral enzymes are very crucial for the inhibition of viruses due to the fact that they are involved in the survival, proliferation and transmission of viruses. The main goal of this study is the presentation of two novel updated phylogenetic trees of the enzymes NS3 helicase and NS5 RdRp as a reliable phylogeny "map" to correlate the information of the closely related viruses and identify new possible targets for the Flaviviridae family virus inhibition. Despite the earliest trials for drugs against Flaviviridae related viruses, no antiviral drug vaccine has been available to date. Therefore there is an urgent need for research towards the development of efficient antiviral agents.

  14. Discovery of Multitarget Antivirals Acting on Both the Dengue Virus NS5-NS3 Interaction and the Host Src/Fyn Kinases.

    PubMed

    Vincetti, Paolo; Caporuscio, Fabiana; Kaptein, Suzanne; Gioiello, Antimo; Mancino, Valentina; Suzuki, Youichi; Yamamoto, Naoki; Crespan, Emmanuele; Lossani, Andrea; Maga, Giovanni; Rastelli, Giulio; Castagnolo, Daniele; Neyts, Johan; Leyssen, Pieter; Costantino, Gabriele; Radi, Marco

    2015-06-25

    This study describes the discovery of novel dengue virus inhibitors targeting both a crucial viral protein-protein interaction and an essential host cell factor as a strategy to reduce the emergence of drug resistance. Starting from known c-Src inhibitors, a virtual screening was performed to identify molecules able to interact with a recently discovered allosteric pocket on the dengue virus NS5 polymerase. The selection of cheap-to-produce scaffolds and the exploration of the biologically relevant chemical space around them suggested promising candidates for chemical synthesis. A series of purines emerged as the most interesting candidates able to inhibit virus replication at low micromolar concentrations with no significant toxicity to the host cell. Among the identified antivirals, compound 16i proved to be 10 times more potent than ribavirin, showed a better selectivity index and represents the first-in-class DENV-NS5 allosteric inhibitor able to target both the virus NS5-NS3 interaction and the host kinases c-Src/Fyn.

  15. Electrochemical lateral flow immunosensor for detection and quantification of dengue NS1 protein.

    PubMed

    Sinawang, Prima Dewi; Rai, Varun; Ionescu, Rodica E; Marks, Robert S

    2016-03-15

    An Electrochemical Lateral Flow Immunosensor (ELFI) is developed combining screen-printed gold electrodes (SPGE) enabling quantification together with the convenience of a lateral flow test strip. A cellulose glassy fiber paper conjugate pad retains the marker immunoelectroactive nanobeads which will bind to the target analyte of interest. The specific immunorecognition event continues to occur along the lateral flow bed until reaching the SPGE-capture antibodies at the end of the cellulosic lateral flow strip. The rationale of the immunoassay consists in the analyte antigen NS1 protein being captured selectively and specifically by the dengue NS1 antibody conjugated onto the immunonanobeads thus forming an immunocomplex. With the aid of a running buffer, the immunocomplexes flow and reach the immuno-conjugated electrode surface and form specific sandwich-type detection due to specific, molecular recognition, while unbound beads move along past the electrodes. The successful sandwich immunocomplex formation is then recorded electrochemically. Specific detection of NS1 is translated into an electrochemical signal contributed by a redox label present on the bead-immobilized detection dengue NS1 antibody while a proportional increase of faradic current is observed with increase in analyte NS1 protein concentration. The first generation ELFI prototype is simply assembled in a cassette and successfully demonstrates wide linear range over a concentration range of 1-25 ng/mL with an ultrasensitive detection limit of 0.5 ng/mL for the qualitative and quantitative detection of analyte dengue NS1 protein.

  16. Characterization of Pressure Transients Generated by Nanosecond Electrical Pulse (nsEP) Exposure

    PubMed Central

    Roth, Caleb C.; Barnes Jr., Ronald A.; Ibey, Bennett L.; Beier, Hope T.; Christopher Mimun, L.; Maswadi, Saher M.; Shadaram, Mehdi; Glickman, Randolph D.

    2015-01-01

    The mechanism(s) responsible for the breakdown (nanoporation) of cell plasma membranes after nanosecond pulse (nsEP) exposure remains poorly understood. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. However, the delivery of a high-voltage nsEP to cells by tungsten electrodes creates a multitude of biophysical phenomena, including electrohydraulic cavitation, electrochemical interactions, thermoelastic expansion, and others. To date, very limited research has investigated non-electric phenomena occurring during nsEP exposures and their potential effect on cell nanoporation. Of primary interest is the production of acoustic shock waves during nsEP exposure, as it is known that acoustic shock waves can cause membrane poration (sonoporation). Based on these observations, our group characterized the acoustic pressure transients generated by nsEP and determined if such transients played any role in nanoporation. In this paper, we show that nsEP exposures, equivalent to those used in cellular studies, are capable of generating high-frequency (2.5 MHz), high-intensity (>13 kPa) pressure transients. Using confocal microscopy to measure cell uptake of YO-PRO®-1 (indicator of nanoporation of the plasma membrane) and changing the electrode geometry, we determined that acoustic waves alone are not responsible for poration of the membrane. PMID:26450165

  17. Amino Terminal Region of Dengue Virus NS4A Cytosolic Domain Binds to Highly Curved Liposomes

    PubMed Central

    Hung, Yu-Fu; Schwarten, Melanie; Hoffmann, Silke; Willbold, Dieter; Sklan, Ella H.; Koenig, Bernd W.

    2015-01-01

    Dengue virus (DENV) is an important human pathogen causing millions of disease cases and thousands of deaths worldwide. Non-structural protein 4A (NS4A) is a vital component of the viral replication complex (RC) and plays a major role in the formation of host cell membrane-derived structures that provide a scaffold for replication. The N-terminal cytoplasmic region of NS4A(1–48) is known to preferentially interact with highly curved membranes. Here, we provide experimental evidence for the stable binding of NS4A(1–48) to small liposomes using a liposome floatation assay and identify the lipid binding sequence by NMR spectroscopy. Mutations L6E;M10E were previously shown to inhibit DENV replication and to interfere with the binding of NS4A(1–48) to small liposomes. Our results provide new details on the interaction of the N-terminal region of NS4A with membranes and will prompt studies of the functional relevance of the curvature sensitive membrane anchor at the N-terminus of NS4A. PMID:26197333

  18. Characterization of Pressure Transients Generated by Nanosecond Electrical Pulse (nsEP) Exposure.

    PubMed

    Roth, Caleb C; Barnes, Ronald A; Ibey, Bennett L; Beier, Hope T; Christopher Mimun, L; Maswadi, Saher M; Shadaram, Mehdi; Glickman, Randolph D

    2015-10-09

    The mechanism(s) responsible for the breakdown (nanoporation) of cell plasma membranes after nanosecond pulse (nsEP) exposure remains poorly understood. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. However, the delivery of a high-voltage nsEP to cells by tungsten electrodes creates a multitude of biophysical phenomena, including electrohydraulic cavitation, electrochemical interactions, thermoelastic expansion, and others. To date, very limited research has investigated non-electric phenomena occurring during nsEP exposures and their potential effect on cell nanoporation. Of primary interest is the production of acoustic shock waves during nsEP exposure, as it is known that acoustic shock waves can cause membrane poration (sonoporation). Based on these observations, our group characterized the acoustic pressure transients generated by nsEP and determined if such transients played any role in nanoporation. In this paper, we show that nsEP exposures, equivalent to those used in cellular studies, are capable of generating high-frequency (2.5 MHz), high-intensity (>13 kPa) pressure transients. Using confocal microscopy to measure cell uptake of YO-PRO®-1 (indicator of nanoporation of the plasma membrane) and changing the electrode geometry, we determined that acoustic waves alone are not responsible for poration of the membrane.

  19. Avian and mammalian reoviruses use different molecular mechanisms to synthesize their {micro}NS isoforms.

    PubMed

    Busch, Lisa K; Rodríguez-Grille, Javier; Casal, J Ignacio; Martínez-Costas, José; Benavente, Javier

    2011-11-01

    Previous reports revealed that the M3 gene of both avian and mammalian reoviruses express two isoforms of the non-structural protein μNS in infected cells. The larger isoforms initiate translation at the AUG codon closest to the 5' end of their respective m3 mRNAs, and were therefore designated μNS. In this study we have performed experiments to identify the molecular mechanisms by which the smaller μNS isoforms are generated. The results of this study confirmed the previous findings indicating that the smaller mammalian reovirus μNS isoform is a primary translation product, the translation of which is initiated at the internal AUG-41 codon of mammalian reovirus m3 mRNA. Our results further revealed that the smaller avian reovirus μNS isoform originates from a specific post-translational cleavage site near the amino terminus of μNS. This cleavage produces a 55 kDa carboxy-terminal protein, termed μNSC, and a 17 kDa amino-terminal polypeptide, designated μNSN. These results allowed us to extend the known avian reovirus protein-encoding capacity to 18 proteins, 12 of which are structural proteins and six of which are non-structural proteins. Our finding that avian and mammalian reoviruses use different mechanisms to express their μNSC isoforms suggests that these isoforms are important for reovirus replication.

  20. Murine coronavirus nonstructural protein ns2 is not essential for virus replication in transformed cells.

    PubMed Central

    Schwarz, B; Routledge, E; Siddell, S G

    1990-01-01

    Two isolates of the murine hepatitis virus (MHV) strain JHM, which differed in their ability to express the nonstructural gene product ns2, were characterized. The MHV Wb3 isolate encodes a 30,000-molecular-weight ns2 protein that can be readily detected in infected cells by using a specific monoclonal antibody, MAb 2A. The MHV Wb1 isolate is a deletion mutant that lacks a functional ns2 gene and the transcriptional signals required for the synthesis of an ns2 mRNA. However, there are no obviously significant differences in the growth of the MHV Wb1 and MHV Wb3 isolates in continuous cell lines or in the synthesis of viral mRNAs or proteins in infected cells. These results demonstrate that the ns2 gene product is not essential for MHV replication in transformed murine cells and suggests that the function of the ns2 gene may only be manifest in vivo. Images PMID:2168966

  1. Characterization of Pressure Transients Generated by Nanosecond Electrical Pulse (nsEP) Exposure

    NASA Astrophysics Data System (ADS)

    Roth, Caleb C.; Barnes, Ronald A., Jr.; Ibey, Bennett L.; Beier, Hope T.; Christopher Mimun, L.; Maswadi, Saher M.; Shadaram, Mehdi; Glickman, Randolph D.

    2015-10-01

    The mechanism(s) responsible for the breakdown (nanoporation) of cell plasma membranes after nanosecond pulse (nsEP) exposure remains poorly understood. Current theories focus exclusively on the electrical field, citing electrostriction, water dipole alignment and/or electrodeformation as the primary mechanisms for pore formation. However, the delivery of a high-voltage nsEP to cells by tungsten electrodes creates a multitude of biophysical phenomena, including electrohydraulic cavitation, electrochemical interactions, thermoelastic expansion, and others. To date, very limited research has investigated non-electric phenomena occurring during nsEP exposures and their potential effect on cell nanoporation. Of primary interest is the production of acoustic shock waves during nsEP exposure, as it is known that acoustic shock waves can cause membrane poration (sonoporation). Based on these observations, our group characterized the acoustic pressure transients generated by nsEP and determined if such transients played any role in nanoporation. In this paper, we show that nsEP exposures, equivalent to those used in cellular studies, are capable of generating high-frequency (2.5 MHz), high-intensity (>13 kPa) pressure transients. Using confocal microscopy to measure cell uptake of YO-PRO®-1 (indicator of nanoporation of the plasma membrane) and changing the electrode geometry, we determined that acoustic waves alone are not responsible for poration of the membrane.

  2. Influenza A Virus NS1 Protein Inhibits the NLRP3 Inflammasome

    PubMed Central

    Cheong, Woo-Chang; Kang, Hye-Ri; Yoon, Hyunyee; Kang, Suk-Jo; Ting, Jenny P.-Y.; Song, Moon Jung

    2015-01-01

    The inflammasome is a molecular platform that stimulates the activation of caspase-1 and the processing of pro-interleukin (IL)-1β and pro-IL-18 for secretion. The NOD-like receptor family, pyrin domain containing 3 (NLRP3) protein is activated by diverse molecules and pathogens, leading to the formation of the NLRP3 inflammasome. Recent studies showed that the NLRP3 inflammasome mediates innate immunity against influenza A virus (IAV) infection. In this study, we investigated the function of the IAV non-structural protein 1 (NS1) in the modulation of NLRP3 inflammasome. We found that NS1 proteins derived from both highly pathogenic and low pathogenic strains efficiently decreased secretion of IL-1β and IL-18 from THP-1 cells treated with LPS and ATP. NS1 overexpression significantly impaired the transcription of proinflammatory cytokines by inhibiting transactivation of the nuclear factor-κB (NF-κB), a major transcription activator. Furthermore, NS1 physically interacted with endogenous NLRP3 and activation of the NLRP3 inflammasome was abrogated in NS1-expressing THP-1 cells. These findings suggest that NS1 downregulates NLRP3 inflammasome activation by targeting NLRP3 as well as NF-κB, leading to a reduction in the levels of inflammatory cytokines as a viral immune evasion strategy. PMID:25978411

  3. Nuclear Translocation of Crk Adaptor Proteins by the Influenza A Virus NS1 Protein

    PubMed Central

    Ylösmäki, Leena; Fagerlund, Riku; Kuisma, Inka; Julkunen, Ilkka; Saksela, Kalle

    2016-01-01

    The non-structural protein-1 (NS1) of many influenza A strains, especially those of avian origin, contains an SH3 ligand motif, which binds tightly to the cellular adaptor proteins Crk (Chicken tumor virus number 10 (CT10) regulator of kinase) and Crk-like adapter protein (CrkL). This interaction has been shown to potentiate NS1-induced activation of the phosphatidylinositol 3-kinase (PI3K), but additional effects on the host cell physiology may exist. Here we show that NS1 can induce an efficient translocation of Crk proteins from the cytoplasm into the nucleus, which results in an altered pattern of nuclear protein tyrosine phosphorylation. This was not observed using NS1 proteins deficient in SH3 binding or engineered to be exclusively cytoplasmic, indicating a physical role for NS1 as a carrier in the nuclear translocation of Crk. These data further emphasize the role of Crk proteins as host cell interaction partners of NS1, and highlight the potential for host cell manipulation gained by a viral protein simply via acquiring a short SH3 binding motif. PMID:27092521

  4. Activation of interferon regulatory factor 3 is inhibited by the influenza A virus NS1 protein.

    PubMed

    Talon, J; Horvath, C M; Polley, R; Basler, C F; Muster, T; Palese, P; García-Sastre, A

    2000-09-01

    We present a novel mechanism by which viruses may inhibit the alpha/beta interferon (IFN-alpha/beta) cascade. The double-stranded RNA (dsRNA) binding protein NS1 of influenza virus is shown to prevent the potent antiviral interferon response by inhibiting the activation of interferon regulatory factor 3 (IRF-3), a key regulator of IFN-alpha/beta gene expression. IRF-3 activation and, as a consequence, IFN-beta mRNA induction are inhibited in wild-type (PR8) influenza virus-infected cells but not in cells infected with an isogenic virus lacking the NS1 gene (delNS1 virus). Furthermore, NS1 is shown to be a general inhibitor of the interferon signaling pathway. Inhibition of IRF-3 activation can be achieved by the expression of wild-type NS1 in trans, not only in delNS1 virus-infected cells but also in cells infected with a heterologous RNA virus (Newcastle disease virus). We propose that inhibition of IRF-3 activation by a dsRNA binding protein significantly contributes to the virulence of influenza A viruses and possibly to that of other viruses.

  5. Flavivirus NS4A-induced autophagy protects cells against death and enhances virus replication.

    PubMed

    McLean, Jeffrey E; Wudzinska, Aleksandra; Datan, Emmanuel; Quaglino, Daniela; Zakeri, Zahra

    2011-06-24

    Flaviviruses include the most prevalent and medically challenging viruses. Persistent infection with flaviviruses of epithelial cells and hepatocytes that do not undergo cell death is common. Here, we report that, in epithelial cells, up-regulation of autophagy following flavivirus infection markedly enhances virus replication and that one flavivirus gene, NS4A, uniquely determines the up-regulation of autophagy. Dengue-2 and Modoc (a murine flavivirus) kill primary murine macrophages but protect epithelial cells and fibroblasts against death provoked by several insults. The flavivirus-induced protection derives from the up-regulation of autophagy, as up-regulation of autophagy by starvation or inactivation of mammalian target of rapamycin also protects the cells against insult, whereas inhibition of autophagy via inactivation of PI3K nullifies the protection conferred by flavivirus. Inhibition of autophagy also limits replication of both Dengue-2 and Modoc virus in epithelial cells. Expression of flavivirus NS4A is sufficient to induce PI3K-dependent autophagy and to protect cells against death; expression of other viral genes, including NS2A and NS4B, fails to protect cells against several stressors. Flavivirus NS4A protein induces autophagy in epithelial cells and thus protects them from death during infection. As autophagy is vital to flavivirus replication in these cells, NS4A is therefore also identified as a critical determinant of flavivirus replication. PMID:21511946

  6. Electrochemical lateral flow immunosensor for detection and quantification of dengue NS1 protein.

    PubMed

    Sinawang, Prima Dewi; Rai, Varun; Ionescu, Rodica E; Marks, Robert S

    2016-03-15

    An Electrochemical Lateral Flow Immunosensor (ELFI) is developed combining screen-printed gold electrodes (SPGE) enabling quantification together with the convenience of a lateral flow test strip. A cellulose glassy fiber paper conjugate pad retains the marker immunoelectroactive nanobeads which will bind to the target analyte of interest. The specific immunorecognition event continues to occur along the lateral flow bed until reaching the SPGE-capture antibodies at the end of the cellulosic lateral flow strip. The rationale of the immunoassay consists in the analyte antigen NS1 protein being captured selectively and specifically by the dengue NS1 antibody conjugated onto the immunonanobeads thus forming an immunocomplex. With the aid of a running buffer, the immunocomplexes flow and reach the immuno-conjugated electrode surface and form specific sandwich-type detection due to specific, molecular recognition, while unbound beads move along past the electrodes. The successful sandwich immunocomplex formation is then recorded electrochemically. Specific detection of NS1 is translated into an electrochemical signal contributed by a redox label present on the bead-immobilized detection dengue NS1 antibody while a proportional increase of faradic current is observed with increase in analyte NS1 protein concentration. The first generation ELFI prototype is simply assembled in a cassette and successfully demonstrates wide linear range over a concentration range of 1-25 ng/mL with an ultrasensitive detection limit of 0.5 ng/mL for the qualitative and quantitative detection of analyte dengue NS1 protein. PMID:26433352

  7. Nonstructural protein 1 characteristic peak from NS1-saliva mixture with Surface-Enhanced Raman spectroscopy.

    PubMed

    Radzol, A R M; Lee, Khuan Y; Mansor, W

    2013-01-01

    Surface Enhanced Raman spectroscopy (SERS) is an enhanced technique of Raman spectroscopy, which amplifies the intensity of Raman scattering to a practical range with adsorption of analyte onto nano-size plasmonic material such as gold, silver or copper. This feature of SERS has given it a niche in tracing molecular structure, especially useful for marking diseases specific biomarker. NS1 protein has been clinically accepted as an alternative biomarker for diseases caused by flavivirus. Detection of Nonstructural Protein 1 (NS1) will allow early diagnosis of the diseases. Its presence in the blood serum has been reported as early as first day of infection. With gold substrate, our work here intends to explore if SERS is suitable to detect NS1 from saliva, with saliva becoming the most favored alternative to blood as diagnostic fluid due to its advantages in sample collection. Our experimental results find both gold coated slide (GS) and saliva being Raman inactive, but the molecular fingerprint of NS1 protein at Raman shift 1012 cm(-1), which has never been reported before. The distinct peak is discovered to be attributed by breathing vibration of the benzene ring structure of NS1 side chain molecule. The characteristic peak is also found to vary in direct proportion to concentration of the NS1-saliva mixture, with a correlation coefficient of +0.96118 and a standard error estimation of 0.11382.

  8. Specific binding of Bluetongue virus NS2 to different viral plus-strand RNAs.

    PubMed

    Lymperopoulos, Kostas; Noad, Rob; Tosi, Sara; Nethisinghe, Suran; Brierley, Ian; Roy, Polly

    2006-09-15

    The Reoviridae have double-stranded RNA genomes of 10-12 segments, each in a single copy in the mature virion. The basis of genome segment sorting during virus assembly that ensures each virus particle contains the complete viral genome is unresolved. Bluetongue virus (BTV) NS2 is a single-stranded RNA-binding protein that forms inclusion bodies in infected cells. Here, we demonstrate that the specific interaction between NS2 and a stem-loop structure present in BTV S10 RNA, and phylogenetically conserved in other BTV serotypes, is abolished by mutations predicted to disrupt the structure. Subsequently, we mapped RNA regions in three other genomic segments of BTV that are bound preferentially by NS2. However, structure probing of these RNAs did not reveal secondary structure motifs that obviously resembled the stem-loop implicated in the NS2-S10 interaction. In addition, the specific binding by NS2 to two different viral RNAs was found to occur independently. Together, these data support the hypothesis that the recognition by NS2 of different RNA structures may be the basis for discrimination between viral RNAs during virus assembly.

  9. Influenza virus subpopulations: exchange of lethal H5N1 virus NS for H1N1 virus NS triggers de novo generation of defective-interfering particles and enhances interferon-inducing particle efficiency.

    PubMed

    Ngunjiri, John M; Buchek, Gregory M; Mohni, Kareem N; Sekellick, Margaret J; Marcus, Philip I

    2013-03-01

    Reassortment of influenza A viruses is known to affect viability, replication efficiency, antigenicity, host range, and virulence, and can generate pandemic strains. In this study, we demonstrated that the specific exchange of the NS gene segment from highly pathogenic A/HK/156/97 (H5N1) [E92 or E92D NS1] virus for the cognate NS gene segment of A/PR/834(H1N1) [D92 NS1] virus did not cause a significant change in the sizes of infectious particle subpopulations. However, it resulted in 2 new phenotypic changes: (1) de novo generation of large subpopulations of defective-interfering particles (DIPs); and (2) enhancement of interferon (IFN)-inducing particle efficiency leading to an order of magnitude or higher quantum (peak) yield of IFN in both avian and mammalian cells. These changes were attributed to loss of function of the H5N1-NS gene products. Most notably, the NS exchange obliterated the usual IFN-induction-suppressing capacity associated with expression of full-size NS1 proteins, and hence functionally mimicked deletions in the NS1 gene. The loss of NS1-mediated suppression of IFN induction, de novo generation of DIPs, and the concomitant enhancement of IFN-inducing particle efficiency suggest that in an attenuated background, the H5N1-NS could be used to formulate a self-adjuvanting live attenuated influenza vaccine similar to viruses with deletions in the NS1 gene.

  10. Extremely Low Passive Microwave Brightness Temperatures Due to Thunderstorms

    NASA Technical Reports Server (NTRS)

    Cecil, Daniel J.

    2015-01-01

    Extreme events by their nature fall outside the bounds of routine experience. With imperfect or ambiguous measuring systems, it is appropriate to question whether an unusual measurement represents an extreme event or is the result of instrument errors or other sources of noise. About three weeks after the Tropical Rainfall Measuring Mission (TRMM) satellite began collecting data in Dec 1997, a thunderstorm was observed over northern Argentina with 85 GHz brightness temperatures below 50 K and 37 GHz brightness temperatures below 70 K (Zipser et al. 2006). These values are well below what had previously been observed from satellite sensors with lower resolution. The 37 GHz brightness temperatures are also well below those measured by TRMM for any other storm in the subsequent 16 years. Without corroborating evidence, it would be natural to suspect a problem with the instrument, or perhaps an irregularity with the platform during the first weeks of the satellite mission. Automated quality control flags or other procedures in retrieval algorithms could treat these measurements as errors, because they fall outside the expected bounds. But the TRMM satellite also carries a radar and a lightning sensor, both confirming the presence of an intense thunderstorm. The radar recorded 40+ dBZ reflectivity up to about 19 km altitude. More than 200 lightning flashes per minute were recorded. That same storm's 19 GHz brightness temperatures below 150 K would normally be interpreted as the result of a low-emissivity water surface (e.g., a lake, or flood waters) if not for the simultaneous measurements of such intense convection. This paper will examine records from TRMM and related satellite sensors including SSMI, AMSR-E, and the new GMI to find the strongest signatures resulting from thunderstorms, and distinguishing those from sources of noise. The lowest brightness temperatures resulting from thunderstorms as seen by TRMM have been in Argentina in November and December. For

  11. Direct Binding of Ledipasvir to HCV NS5A: Mechanism of Resistance to an HCV Antiviral Agent

    PubMed Central

    Kwon, Hyock Joo; Xing, Weimei; Chan, Katie; Niedziela-Majka, Anita; Brendza, Katherine M.; Kirschberg, Thorsten; Kato, Darryl; Link, John O.; Cheng, Guofeng; Liu, Xiaohong; Sakowicz, Roman

    2015-01-01

    Ledipasvir, a direct acting antiviral agent (DAA) targeting the Hepatitis C Virus NS5A protein, exhibits picomolar activity in replicon cells. While its mechanism of action is unclear, mutations that confer resistance to ledipasvir in HCV replicon cells are located in NS5A, suggesting that NS5A is the direct target of ledipasvir. To date co-precipitation and cross-linking experiments in replicon or NS5A transfected cells have not conclusively shown a direct, specific interaction between NS5A and ledipasvir. Using recombinant, full length NS5A, we show that ledipasvir binds directly, with high affinity and specificity, to NS5A. Ledipasvir binding to recombinant NS5A is saturable with a dissociation constant in the low nanomolar range. A mutant form of NS5A (Y93H) that confers resistance to ledipasvir shows diminished binding to ledipasvir. The current study shows that ledipasvir inhibits NS5A through direct binding and that resistance to ledipasvir is the result of a reduction in binding affinity to NS5A mutants. PMID:25856426

  12. Thermal neutron image intensifier tube provides brightly visible radiographic pattern

    NASA Technical Reports Server (NTRS)

    Berger, H.; Kraska, I.; Niklas, W.; Schmidt, A.

    1967-01-01

    Vacuum-type neutron image intensifier tube improves image detection in thermal neutron radiographic inspection. This system converts images to an electron image, and with electron acceleration and demagnification between the input target and output screen, produces a bright image viewed through a closed circuit television system.

  13. High-brightness high-duty cycle electron injectors

    NASA Astrophysics Data System (ADS)

    Sannibale, Fernando

    2014-03-01

    High brightness electron sources have been one of the driving forces behind the spectacular results achieved in the last decade by accelerator-based applications. Indeed, X-Ray FELs, with their 10-fold orders of magnitude increase in peak brightness, probably represent the best example of it. New ambitious proposals for X-ray FELs and ERLs, as well as inverse Compton sources for X- or gamma-ray production, are now requiring operation at MHz-GHz repetition rates. In response to that, a number of groups around the world have been actively working in developing high-brightness high-duty cycle electron injectors capable of driving such machines. The high repetition rate requirement cannot be met by the existing low-repetition rate high-brightness injector schemes, and new technologies are under investigation. This paper includes a description of the requirements for such injectors, an overview of the pursued technologies and schemes, a description of the main beam dynamics issues associated with the different choices, and a number of examples of the results obtained so far by the groups active in the field.

  14. RESPONSES OF BRIGHT, NORMAL, AND RETARDED CHILDREN TO LEARNING TASKS.

    ERIC Educational Resources Information Center

    CARRIER, NEIL A.; AND OTHERS

    THE RELATIONSHIPS AMONG THE VARIABLES OF INTELLIGENCE, LEARNING TASK PERFORMANCE, EMOTIONAL TENSION, AND TASK MOTIVATION WERE STUDIED. ABOUT 120 BRIGHT, NORMAL, AND RETARDED CHILDREN PERFORMED SIX TRIALS OF NUMBER LEARNING, CONCEPT FORMATION, PROBLEM SOLVING, PERCEPTUAL-MOTOR COORDINATION, AND VERBAL LEARNING TASKS. DURING THE LEARNING SESSIONS,…

  15. Apollo experiment S-211 low brightness, astronomical photography

    NASA Technical Reports Server (NTRS)

    Mercer, R. D.

    1974-01-01

    The low brightness image analysis for the Apollo flights is reported. The objectives of Experiment S-21 are discussed along with the photographic equipment, and the data processing and analysis flow are presented. Photographic data for Apollo 14, 15, 16, and 17 are included.

  16. Low surface brightness galaxies and tidally triggered star formation

    NASA Technical Reports Server (NTRS)

    Zaritsky, Dennis; Lorrimer, Stephen J.

    1993-01-01

    Counts of companions to low surface brightness (LSB) galaxies are presented and these are compared to counts of companions to normal galaxies obtained with the same techniques and criteria. Our results are consistent with LSB's having no clustered companions and support the hypothesis that LSB galaxies have low star-formation rates because they lack external tidal triggering.

  17. The "Brightness Rules" Alternative Conception for Light Bulb Circuits

    ERIC Educational Resources Information Center

    Bryan, Joel A.; Stuessy, Carol

    2006-01-01

    An alternative conception for the observed differences in light bulb brightness was revealed during an unguided inquiry investigation in which prospective elementary teachers placed identical bulbs in series, parallel, and combination direct current circuits. Classroom observations, document analyses, and video and audio transcriptions led to the…

  18. HANDS-ON, Career Exploration for Bright Students.

    ERIC Educational Resources Information Center

    Stovall, Betty J.; And Others

    Produced as part of a 5 week workshop on career explorations for 51 bright, middle grade students and 20 teachers, the curriculum guide discusses career education, outlines the workshop experiences, considers the inquiry process, and outlines 60 units on non baccalaureate careers in 15 career clusters. A lack of career education programs with…

  19. Femtosecond, High-Brightness Electron Beam Generation and Advanced Diagnosis

    SciTech Connect

    Anderson, S G; Brown, W J; Tremaine, A M; Kuba, J; Hartemann, F V; Fittinghoff, D N

    2005-02-02

    This document serves as the final report for LDRD project number 04-LW-031, in which we created subpicosecond length, kilo-amp peak current electron beams with the 100 MeV electron/positron linac, using a novel technique designed to produce ultra-short bunch lengths while maintaining the high brightness produced by the S-band photoinjector. In addition, a diagnostic to measure the temporal distribution of the beam was investigated, as conventional pulse length measurement techniques do not apply to extremely short pulses. The creation and diagnosis of beams with both femtosecond length and high transverse brightness is of major concern to next generation acceleration and radiation production experiments. This work leveraged the previous investment in the PLEIADES facility and it's ability to produce high brightness electron beams. In addition, the ultra-short electron pulses generated by this work have been used in conjunction with the PLEIADES X-ray source to produce sub-picosecond, high-brightness X-ray pulses.

  20. High Precision Photometry of Bright Transiting Exoplanet Hosts

    NASA Astrophysics Data System (ADS)

    Wilson, Maurice; Eastman, Jason; Johnson, John A.

    2016-01-01

    Within the past two decades, the successful search for exoplanets and the characterization of their physical properties have shown the immense progress that has been made towards finding planets with characteristics similar to Earth. For most exoplanets with a radius about the size of Earth, evaluating their physical properties, such as the mass, radius and equilibrium temperature, cannot be determined with satisfactory precision. The MINiature Exoplanet Radial Velocity Array (MINERVA) was recently built to obtain spectroscopic and photometric measurements to find, confirm, and characterize Earth-like exoplanets. MINERVA's spectroscopic survey targets the brightest, nearby stars which are well-suited to the array's capabilities, while its primary photometric goal is to search for transits around these bright targets. Typically, it is difficult to find satisfactory comparison stars within a telescope's field of view when the primary target is very bright. This issue is resolved by using one of MINERVA's telescopes to observe the primary bright star while the other telescopes observe a distinct field of view that contains satisfactory bright comparison stars. We describe the code used to identify nearby comparison stars, schedule the four telescopes, produce differential photometry from multiple telescopes, and show the first results from this effort.This work has been funded by the Ronald E. McNair Post-Baccalaureate Achievement Program, the ERAU Honors Program, the ERAU Undergraduate Research Spark Fund, and the Banneker Institute at the Harvard-Smithsonian Center for Astrophysics.

  1. Exploring the Multi-Wavelength, Low Surface Brightness Universe

    NASA Astrophysics Data System (ADS)

    Brunner, R. J.; Djorgovski, S. G.; Gal, R. R.; Odewahn, S. C.

    1999-12-01

    Our current understanding of the low surface brightness universe is quite incomplete, not only in the optical, but also in other wavelength regimes. We have, therefore, begun an exploration of the multi-wavelength, low surface brightness universe. This project currently involves data from DPOSS (Digitized Palomar Optical Sky Survey), 2MASS (Two Micron All Sky Survey), IRAS (Infrared Astronomical Satellite), and several neutral hydrogen surveys. We present some initial results of this work as well as discuss the implications of this work on future virtual observatories. Our main scientific goals have been the search for low surface brightness galaxies, including local group dwarf spheroidals, and also optical counterparts to high velocity clouds. Our techniques are complimentary to normal data reduction pipeline techniques in that we focus on the diffuse emission that is ignored or removed by more traditional algorithms. This requires, of course, a spatial filtering which must account for objects of interest, in addition to observational artifacts (e.g.,\\ bright stellar halos). Finally, with this work, we are exploring the intersection of the catalog and image domains in order to maximize the scientific information we can extract from the federation of large survey data.

  2. Impact Evaluation of Burkina Faso's BRIGHT Program. Final Report

    ERIC Educational Resources Information Center

    Levy, Dan; Sloan, Matt; Linden, Leigh; Kazianga, Harounan

    2009-01-01

    The BRIGHT program was designed to improve the educational outcomes of children in Burkina Faso. It focused on girls in particular and was implemented in 132 rural villages throughout the 10 provinces of the country in which girls' enrollment rates were lowest. It consisted of constructing primary schools with three classrooms and implementing a…

  3. Aviation -- Where Career Opportunities are Bright, Counselor's Guide.

    ERIC Educational Resources Information Center

    Zaharevitz, Walter; Marshall, Jane N.

    This aviation occupations guide is designed for use as a unit as well as in conjunction with an aviation careers package of material that contains a film strip and recording. Chapter One contains the script of the film strip, Aviation--Where Career Opportunities are Bright, and includes all photographs used in the film strip plus numerous…

  4. Reducing Color/Brightness Interaction in Color Television

    NASA Technical Reports Server (NTRS)

    Marchman, Robert H.

    1987-01-01

    Proposed digitally sampled scan-conversion scheme for color television reduces unwanted interactions between chrominance and luminance signals. New scheme reduces luminance and chrominance bandwidth to increase frequency separation between signals. To avoid proportionally reducing horizontal brightness resolution and horizontal color resolution, horizontal interlace of luminance signal and two color-difference signals used.

  5. Finding Bright-Spot Coordinates in Television Images

    NASA Technical Reports Server (NTRS)

    Richardson, T. E.; Tietz, J. C.

    1985-01-01

    Circuit provides data for computer to calculate coordinates of bright spot of light in video image. Calculation performed while image being scanned, and results available immediately at end of video frame. Video-processing circuit has variety of potential uses in commerce and industry. For example, locates tagged-parts on factory assembly line or track airplane landing lights.

  6. Apparatus Would Position Bright Spot On Projection Screen

    NASA Technical Reports Server (NTRS)

    Rayman, Marc D.

    1996-01-01

    Proposed apparatus aims beam of visible light at wavelength lambda(2) to create bright spot at desired position in image on projection screen. Intended to replace handheld laser and flashlight pointers lecturers sometimes use to indicate features in projected images. Beam of light cannot be inadvertently aimed toward audience.

  7. Variations in the Fe mineralogy of bright Martian soil

    NASA Technical Reports Server (NTRS)

    Murchie, Scott; Mustard, John; Erard, Stephane; Geissler, Paul; Singer, Robert

    1993-01-01

    Bright regions on Mars are interpreted as 'soil' derived by chemical alteration of crustal rocks, whose main pigmentary component is ferric oxide or oxyhydroxide. The mineralogy and mineralogic variability of ferric iron are important evidence for the evolution of Martian soil: mineralogy of ferric phases is sensitive to chemical conditions in their genetic environments, and the spatial distributions of different ferric phases would record a history of both chemical environments and physical mixing. Reflectance spectroscopic studies provide several types of evidence that discriminate possible pigmentary phases, including the position of a crystal field absorption near 0.9 microns and position and strengths of absorptions in the UV-visible wavelength region. Recent telescopic spectra and laboratory measurements of Mars soil analogs suggest that spectral features of bright soil can be explained based on a single pigmentary phase, hematite (alpha-Fe2O3), occurring in both 'nanophase' and more crystalline forms. Here we report on a systematic investigation of Martian bright regions using ISM imaging spectrometer data, in which we examined spatial variations in the position and shape of the approximately 0.9 microns absorption. We found both local and regional heterogeneities that indicate differences in Fe mineralogy. These results demonstrate that bright soils do not represent a single lithology that has been homogenized by eolian mixing, and suggest that weathering of soils in different geologic settings has followed different physical and chemical pathways.

  8. Soft-X-Ray Prefilter for Hot, Bright Objects

    NASA Technical Reports Server (NTRS)

    Davis, J. M.; Ortendahl, J. A.

    1985-01-01

    Prefilters consisting of beryllium foil supported on conductive silver mesh transmit soft x-rays but are nearly opaque to visible and infrared light. New Be/AG filters protect imaging X-ray detectors from damage by visible and longer wavelength radiation when viewing such hot, bright emitters as Sun or possibly certain industrial processes.

  9. Properties of lateral interaction in color and brightness induction.

    PubMed

    Bachy, Romain; Zaidi, Qasim

    2016-03-01

    In a visual scene, when objects are surrounded by other components, neural mechanisms increase the perceived color and brightness difference between an object and its surround, potentially enhancing an observer's ability to segment objects. Despite almost two centuries of empirical investigations, the nature of induction mechanisms remains elusive. To elucidate the nature of these mechanisms, we introduce a new method for measuring color and brightness induction that allows separate manipulation of lateral interactions and adaptation, and controls for eye-movement-related effects. We use the method to examine the function relating induction magnitude to contrast change in the surround, the symmetry of induction in complementary directions for the three cardinal color axes, and the effect of blur between the test and the surround. On average, brightness induction was more linear than chromatic induction. The induction magnitude was similar for surrounds of complementary colors on average and for many conditions, and when individual observers deviated from symmetry it could be on either side. Edge blur did not change the induction magnitude. For slower presentations, light/dark induction increased to further reduce asymmetry, suggesting that previously found light/dark induction asymmetry is not due to lateral interactions or prolonged adaptation. Lateral interactions underlying induction are thus mostly symmetric for color and brightness axes and involve spatially opponent filters of modest widths, rather than edge extraction.

  10. Bright Sneezes and Dark Coughs, Loud Sunlight and Soft Moonlight.

    ERIC Educational Resources Information Center

    Marks, Lawrence E.

    1982-01-01

    In a series of four experiments, subjects used scales of loudness, pitch, and brightness to evaluate the meanings of a variety of synesthetic metaphors--expressions in which words or phrases describing experiences proper to one sense modality transfer their meaning to another modality. (Author/PN)

  11. The 1997 Reference of Diffuse Night Sky Brightness

    NASA Technical Reports Server (NTRS)

    Leinert, C.; Bowyer, S.; Haikala, L. K.; Hanner, M. S.; Hauser, M. G.; Levasseur-Regourd, A. C.; Mann, I.; Mattila, K.; Reach, W. T.; Schlosser, W.; Staude, J. J.; Toller, G. N.; Weiland, J. L.; Weinberg, J. L.; Witt, A. N.

    1997-01-01

    In the following we present material in tabular and graphical form, with the aim to allow the non specialist to obtain a realistic estimate of the diffuse night sky brightness over a wide range of wavelengths from the far UV longward of Ly to the far-infrared.

  12. Soil Moisture Active/Passive (SMAP) Forward Brightness Temperature Simulator

    NASA Technical Reports Server (NTRS)

    Peng, Jinzheng; Peipmeier, Jeffrey; Kim, Edward

    2012-01-01

    The SMAP is one of four first-tier missions recommended by the US National Research Council's Committee on Earth Science and Applications from Space (Earth Science and Applications from Space: National Imperatives for the Next Decade and Beyond, Space Studies Board, National Academies Press, 2007) [1]. It is to measure the global soil moisture and freeze/thaw from space. One of the spaceborne instruments is an L-band radiometer with a shared single feedhorn and parabolic mesh reflector. While the radiometer measures the emission over a footprint of interest, unwanted emissions are also received by the antenna through the antenna sidelobes from the cosmic background and other error sources such as the Sun, the Moon and the galaxy. Their effects need to be considered accurately, and the analysis of the overall performance of the radiometer requires end-to-end performance simulation from Earth emission to antenna brightness temperature, such as the global simulation of L-band brightness temperature simulation over land and sea [2]. To assist with the SMAP radiometer level 1B algorithm development, the SMAP forward brightness temperature simulator is developed by adapting the Aquarius simulator [2] with necessary modifications. This poster presents the current status of the SMAP forward brightness simulator s development including incorporating the land microwave emission model and its input datasets, and a simplified atmospheric radiative transfer model. The latest simulation results are also presented to demonstrate the ability of supporting the SMAP L1B algorithm development.

  13. Book Bait V: Using Books to Lure Bright Students.

    ERIC Educational Resources Information Center

    Schlichter, Carol L.

    1992-01-01

    This article provides bibliographic information and annotations for 13 folk tales, fables, and spoofs of folk tales. The items are felt to appeal to the quick wit of bright young students, and the adaptations of tales to other times and places acquaint students with history and culture. (JDD)

  14. Protective immunity to Japanese encephalitis virus associated with anti-NS1 antibodies in a mouse model

    PubMed Central

    2012-01-01

    Background Japanese encephalitis virus (JEV) is a major mosquito-borne pathogen that causes viral encephalitis throughout Asia. Vaccination with an inactive JEV particle or attenuated virus is an efficient preventative measure for controlling infection. Flavivirus NS1 protein is a glycoprotein secreted during viral replication that plays multiple roles in the viral life cycle and pathogenesis. Utilizing JEV NS1 as an antigen in viral vectors induces a limited protective immune response against infection. Previous studies using E. coli-expressed JEV NS1 to immunize mice induced protection against lethal challenge; however, the protection mechanism through cellular and humoral immune responses was not described. Results JEV NS1 was expressed in and purified from Drosophila S2 cells in a native glycosylated multimeric form, which induced T-cell and antibody responses in immunized C3H/HeN mice. Mice vaccinated with 1 μg NS1 with or without water-in-oil adjuvant were partially protected against viral challenge and higher protection was observed in mice with higher antibody titers. IgG1 was preferentially elicited by an adjuvanted NS1 protein, whereas a larger load of IFN-γ was produced in splenocytes from mice immunized with aqueous NS1. Mice that passively received anti-NS1 mouse polyclonal immune sera were protected, and this phenomenon was dose-dependent, whereas protection was low or delayed after the passive transfer of anti-NS1 MAbs. Conclusion The purified NS1 subunit induced protective immunity in relation with anti-NS1 IgG1 antibodies. NS1 protein efficiently stimulated Th1-cell proliferation and IFN-γ production. Protection against lethal challenge was elicited by passive transfer of anti-NS1 antisera, suggesting that anti-NS1 antibodies play a substantial role in anti-viral immunity PMID:22828206

  15. Hepatitis C virus NS3 helicase forms oligomeric structures that exhibit optimal DNA unwinding activity in vitro.

    PubMed

    Sikora, Bartek; Chen, Yingfeng; Lichti, Cheryl F; Harrison, Melody K; Jennings, Thomas A; Tang, Yong; Tackett, Alan J; Jordan, John B; Sakon, Joshua; Cameron, Craig E; Raney, Kevin D

    2008-04-25

    HCV NS3 helicase exhibits activity toward DNA and RNA substrates. The DNA helicase activity of NS3 has been proposed to be optimal when multiple NS3 molecules are bound to the same substrate molecule. NS3 catalyzes little or no measurable DNA unwinding under single cycle conditions in which the concentration of substrate exceeds the concentration of enzyme by 5-fold. However, when NS3 (100 nm) is equimolar with the substrate, a small burst amplitude of approximately 8 nm is observed. The burst amplitude increases as the enzyme concentration increases, consistent with the idea that multiple molecules are needed for optimal unwinding. Protein-protein interactions may facilitate optimal activity, so the oligomeric properties of the enzyme were investigated. Chemical cross-linking indicates that full-length NS3 forms higher order oligomers much more readily than the NS3 helicase domain. Dynamic light scattering indicates that full-length NS3 exists as an oligomer, whereas NS3 helicase domain exists in a monomeric form in solution. Size exclusion chromatography also indicates that full-length NS3 behaves as an oligomer in solution, whereas the NS3 helicase domain behaves as a monomer. When NS3 was passed through a small pore filter capable of removing protein aggregates, greater than 95% of the protein and the DNA unwinding activity was removed from solution. In contrast, only approximately 10% of NS3 helicase domain and approximately 20% of the associated DNA unwinding activity was removed from solution after passage through the small pore filter. The results indicate that the optimally active form of full-length NS3 is part of an oligomeric species in vitro.

  16. Actionable Recommendations in the Bright Futures Child Health Supervision Guidelines

    PubMed Central

    Stanton, J.L.; Downs, S.M.

    2014-01-01

    Summary Background With the growing use of electronic health record systems, there is a demand for an electronic version of the leading American pediatric preventive care guideline, Bright Futures. As computer implementation requires actionable recommendations, it is important to assess to what degree Bright Futures meets criteria for actionability. Objectives We aimed to 1) determine the number of actionable recommendations in the current edition of Bright Futures and 2) to recommend a specific format for representing an important class of guidelines in a way that better facilitates computer implementation. Methods We consolidated all action statements in Bright Futures into recommendations. We then used two dimensions (decidability and executability) in the Guideline Implementability Appraisal v 2.0 (GLIA) to determine the actionability of the recommendations. Decidability means the recommendation states precisely under what conditions to perform those actions. Executability means actions are stated specifically, unambiguously and in sufficient detail. The results were presented in a figure titled Service Interval Diagram (SID), describing actionable recommendations, age intervals during which they are applicable, and how frequently they should occur in that interval. Results We consolidated 2161 action items into 245 recommendations and identified 52 that were actionable (21%). Almost exclusively, these recommendations addressed screening, such as newborn metabolic screening, or child safety, such as car seat use. A limited number (n=13) of recommendations for other areas of anticipatory guidance were also actionable. No recommendations on child discipline, family function or mental health met our criteria for actionability. The SID representing these recommendations is presented in a figure. Conclusion Only a portion of the Bright Futures Guidelines meets criteria for actionability. Substantial work lies ahead to develop most recommendations for anticipatory

  17. Dynamic resetting of the human circadian pacemaker by intermittent bright light

    NASA Technical Reports Server (NTRS)

    Rimmer, D. W.; Boivin, D. B.; Shanahan, T. L.; Kronauer, R. E.; Duffy, J. F.; Czeisler, C. A.

    2000-01-01

    In humans, experimental studies of circadian resetting typically have been limited to lengthy episodes of exposure to continuous bright light. To evaluate the time course of the human endogenous circadian pacemaker's resetting response to brief episodes of intermittent bright light, we studied 16 subjects assigned to one of two intermittent lighting conditions in which the subjects were presented with intermittent episodes of bright-light exposure at 25- or 90-min intervals. The effective duration of bright-light exposure was 31% or 63% compared with a continuous 5-h bright-light stimulus. Exposure to intermittent bright light elicited almost as great a resetting response compared with 5 h of continuous bright light. We conclude that exposure to intermittent bright light produces robust phase shifts of the endogenous circadian pacemaker. Furthermore, these results demonstrate that humans, like other species, exhibit an enhanced sensitivity to the initial minutes of bright-light exposure.

  18. Intra-fiber mode combining schemes, demonstrating high power brightness preservation and coherent-coupling brightness enhancement

    NASA Astrophysics Data System (ADS)

    Shamir, Yariv; Zuitlin, Roey; Glick, Yaakov; Aviel, Matitya; Shafir, Noam; Feldman, Revital; Dahan, Asaf; Urbach, Benayahu; Levy, Daniel; Shekel, Eyal; Sintov, Yoav

    2014-02-01

    We demonstrate intra-fiber couplers performance that is close to complete brightness preservation up to 3kW. Furthermore, when mutually coherent sources were used, the same couplers were able to achieve brightness enhancement with almost no beam quality (BQ) deterioration. The couplers are based on an adiabatic, all-fiber, mode coupling device preserving the lowest spatial mode orders. Brightness levels that approach the theoretical limits were achieved by compressing the participating modes into a tight cross section. Incoherent combination is shown for 2×1, 3×1 and 7×1 combined elements. Additionally, we present a solution for preserving the beam propagation factor of the coupler by using a specialty engineered core delivery fiber. The fabricated components are fully fiber- integrated, hence, without free-space limitations. An overall transmission of <90% was obtained, while the coupler-delivery connection is responsible for less than 0.5% loss. Consequently, relatively low temperatures were observed in the combiner package. Alternatively, utilizing two mutually coherent sources, a quadratic brightness factor improvement was demonstrated. The scheme does not require polarization preserving fibers, and achieved rugged 'in-phase' mode-locking. This allows for a significantly simplified scheme, compared to common coherent combining methods. Prospect on future trends relating to nonlinearities and thermal load management are discussed.

  19. Virulence, Speciation and Antibiotic Susceptibility of Ocular Coagualase Negative Staphylococci (CoNS)

    PubMed Central

    Priya, Ravindran; Mythili, Arumugam; Singh, Yendremban Randhir Babu; Sreekumar, Haridas; Manikandan, Palanisamy; Panneerselvam, Kanesan

    2014-01-01

    Background: Coagulase negative Staphylococci (CoNS) are common inhabitants of human skin and mucous membranes. With the emergence of these organisms as prominent pathogens in patients with ocular infections, investigation has intensified in an effort to identify important virulence factors and to inform new approaches to treatment and prevention. Aim: To isolate CoNS from ocular specimens; to study the possible virulence factors; speciation of coagulase negative staphylococci (CoNS) which were isolated from ocular complications; antibiotic susceptibility testing of ocular CoNS. Materials and Methods: The specimens were collected from the target patients who attended the Microbiology Laboratory of a tertiary care eye hospital in Coimbatore, Tamilnadu state, India. The isolates were subjected to tube and slide coagulase tests for the identification of CoNS. All the isolates were subjected to screening for lipase and protease activities. Screening for other virulence factors viz., slime production on Congo red agar medium and haemagglutination assay with use of 96-well microtitre plates. These isolates were identified upto species level by performing biochemical tests such as phosphatase test, arginine test, maltose and trehalose fermentation tests and novobiocin sensitivity test. The isolates were subjected to antibiotic susceptibility studies, based on the revised standards of Clinical and Laboratory Standards Institutes (CLSI). Results: During the one year of study, among the total 260 individuals who were screened, 100 isolates of CoNS were obtained. Lipolytic activity was seen in all the isolates, whereas 38 isolates showed a positive result for protease. A total of 63 isolates showed slime production. Of 100 isolates, 30 isolates were analyzed for haemagglutination, where 4 isolates showed the capacity to agglutinate the erythrocytes. The results of the biochemical analysis revealed that of the 100 isolates of CoNS, 43% were Staphylococcus epidermidis. The other

  20. Quantification of NS1 dengue biomarker in serum via optomagnetic nanocluster detection

    PubMed Central

    Antunes, Paula; Watterson, Daniel; Parmvi, Mattias; Burger, Robert; Boisen, Anja; Young, Paul; Cooper, Matthew A.; Hansen, Mikkel F.; Ranzoni, Andrea; Donolato, Marco

    2015-01-01

    Dengue is a tropical vector-borne disease without cure or vaccine that progressively spreads into regions with temperate climates. Diagnostic tools amenable to resource-limited settings would be highly valuable for epidemiologic control and containment during outbreaks. Here, we present a novel low-cost automated biosensing platform for detection of dengue fever biomarker NS1 and demonstrate it on NS1 spiked in human serum. Magnetic nanoparticles (MNPs) are coated with high-affinity monoclonal antibodies against NS1 via bio-orthogonal Cu-free ‘click’ chemistry on an anti-fouling surface molecular architecture. The presence of the target antigen NS1 triggers MNP agglutination and the formation of nanoclusters with rapid kinetics enhanced by external magnetic actuation. The amount and size of the nanoclusters correlate with the target concentration and can be quantified using an optomagnetic readout method. The resulting automated dengue fever assay takes just 8 minutes, requires 6 μL of serum sample and shows a limit of detection of 25 ng/mL with an upper detection range of 20000 ng/mL. The technology holds a great potential to be applied to NS1 detection in patient samples. As the assay is implemented on a low-cost microfluidic disc the platform is suited for further expansion to multiplexed detection of a wide panel of biomarkers. PMID:26536916

  1. Quantification of NS1 dengue biomarker in serum via optomagnetic nanocluster detection

    NASA Astrophysics Data System (ADS)

    Antunes, Paula; Watterson, Daniel; Parmvi, Mattias; Burger, Robert; Boisen, Anja; Young, Paul; Cooper, Matthew A.; Hansen, Mikkel F.; Ranzoni, Andrea; Donolato, Marco

    2015-11-01

    Dengue is a tropical vector-borne disease without cure or vaccine that progressively spreads into regions with temperate climates. Diagnostic tools amenable to resource-limited settings would be highly valuable for epidemiologic control and containment during outbreaks. Here, we present a novel low-cost automated biosensing platform for detection of dengue fever biomarker NS1 and demonstrate it on NS1 spiked in human serum. Magnetic nanoparticles (MNPs) are coated with high-affinity monoclonal antibodies against NS1 via bio-orthogonal Cu-free ‘click’ chemistry on an anti-fouling surface molecular architecture. The presence of the target antigen NS1 triggers MNP agglutination and the formation of nanoclusters with rapid kinetics enhanced by external magnetic actuation. The amount and size of the nanoclusters correlate with the target concentration and can be quantified using an optomagnetic readout method. The resulting automated dengue fever assay takes just 8 minutes, requires 6 μL of serum sample and shows a limit of detection of 25 ng/mL with an upper detection range of 20000 ng/mL. The technology holds a great potential to be applied to NS1 detection in patient samples. As the assay is implemented on a low-cost microfluidic disc the platform is suited for further expansion to multiplexed detection of a wide panel of biomarkers.

  2. Quantification of NS1 dengue biomarker in serum via optomagnetic nanocluster detection.

    PubMed

    Antunes, Paula; Watterson, Daniel; Parmvi, Mattias; Burger, Robert; Boisen, Anja; Young, Paul; Cooper, Matthew A; Hansen, Mikkel F; Ranzoni, Andrea; Donolato, Marco

    2015-01-01

    Dengue is a tropical vector-borne disease without cure or vaccine that progressively spreads into regions with temperate climates. Diagnostic tools amenable to resource-limited settings would be highly valuable for epidemiologic control and containment during outbreaks. Here, we present a novel low-cost automated biosensing platform for detection of dengue fever biomarker NS1 and demonstrate it on NS1 spiked in human serum. Magnetic nanoparticles (MNPs) are coated with high-affinity monoclonal antibodies against NS1 via bio-orthogonal Cu-free 'click' chemistry on an anti-fouling surface molecular architecture. The presence of the target antigen NS1 triggers MNP agglutination and the formation of nanoclusters with rapid kinetics enhanced by external magnetic actuation. The amount and size of the nanoclusters correlate with the target concentration and can be quantified using an optomagnetic readout method. The resulting automated dengue fever assay takes just 8 minutes, requires 6 μL of serum sample and shows a limit of detection of 25 ng/mL with an upper detection range of 20000 ng/mL. The technology holds a great potential to be applied to NS1 detection in patient samples. As the assay is implemented on a low-cost microfluidic disc the platform is suited for further expansion to multiplexed detection of a wide panel of biomarkers. PMID:26536916

  3. Characterization of methicillin-resistant coagulase-negative staphylococci (MRCoNS) in retail meat.

    PubMed

    Bhargava, Kanika; Zhang, Yifan

    2014-09-01

    This study was to understand the extent of methicillin-resistant coagulase-negative staphylococci (MRCoNS) serving as a mecA reservoir in retail meat. MRCoNS were isolated from retail meat (beef, chicken, and turkey) in Detroit and characterized by sodA gene sequencing for species identification, staphylococcal cassette chromosome mec (SCCmec) typing, and pulsed-field gel electrophoresis (PFGE). Unique MRCoNS isolates recovered from 25 meat samples were comprised of Staphylococcus sciuri (n = 13), Staphylococcus fleuretti (n = 4), Staphylococcus lentus (n = 3), Staphylococcus epidermidis (n = 2), Staphylococcus vitulinus (n = 1), Staphylococcus saprophyticus (n = 1) and Staphylococcus pasteuri (n = 1). Heterogeneous and composite SCCmec types, including I, III, IV, V, I + V and III + V were identified in 16 isolates. Same SCCmec types were recovered in different staphylococcal species and meat sources. Indistinguishable PFGE patterns were also observed in S. sciuri isolated from beef, chicken, and turkey, and with different SCCmec types. In conclusion, multiple CoNS species can serve as reservoirs for mecA. In addition to the clonal transmission of MRCoNS in meat, horizontal occurrence of SCCmec is observed in staphylococcal species.

  4. RSV-encoded NS2 promotes epithelial cell shedding and distal airway obstruction

    PubMed Central

    Liesman, Rachael M.; Buchholz, Ursula J.; Luongo, Cindy L.; Yang, Lijuan; Proia, Alan D.; DeVincenzo, John P.; Collins, Peter L.; Pickles, Raymond J.

    2014-01-01

    Respiratory syncytial virus (RSV) infection is the major cause of bronchiolitis in young children. The factors that contribute to the increased propensity of RSV-induced distal airway disease compared with other commonly encountered respiratory viruses remain unclear. Here, we identified the RSV-encoded nonstructural 2 (NS2) protein as a viral genetic determinant for initiating RSV-induced distal airway obstruction. Infection of human cartilaginous airway epithelium (HAE) and a hamster model of disease with recombinant respiratory viruses revealed that NS2 promotes shedding of infected epithelial cells, resulting in two consequences of virus infection. First, epithelial cell shedding accelerated the reduction of virus titers, presumably by clearing virus-infected cells from airway mucosa. Second, epithelial cells shedding into the narrow-diameter bronchiolar airway lumens resulted in rapid accumulation of detached, pleomorphic epithelial cells, leading to acute distal airway obstruction. Together, these data indicate that RSV infection of the airway epithelium, via the action of NS2, promotes epithelial cell shedding, which not only accelerates viral clearance but also contributes to acute obstruction of the distal airways. Our results identify RSV NS2 as a contributing factor for the enhanced propensity of RSV to cause severe airway disease in young children and suggest NS2 as a potential therapeutic target for reducing the severity of distal airway disease. PMID:24713657

  5. Analysis of the PDZ binding specificities of Influenza A virus NS1 proteins.

    PubMed

    Thomas, Miranda; Kranjec, Christian; Nagasaka, Kazunori; Matlashewski, Greg; Banks, Lawrence

    2011-01-01

    The Influenza A virus non-structural protein 1 (NS1) is a multifunctional virulence factor with several protein-protein interaction domains, involved in preventing apoptosis of the infected cell and in evading the interferon response. In addition, the majority of influenza A virus NS1 proteins have a class I PDZ-binding motif at the C-terminus, and this itself has been shown to be a virulence determinant.In the majority of human influenza NS1 proteins the consensus motif is RSxV: in avian NS1 it is ESxV. Of the few human strains that have the avian motif, all were from very high mortality outbreaks of the disease. Previous work has shown that minor differences in PDZ-binding motifs can have major effects on the spectrum of cellular proteins targeted. In this study we analyse the effect of these differences upon the binding of Influenza A virus NS1 protein to a range of cellular proteins involved in polarity and signal transduction.

  6. Flavivirus NS1 protein in infected host sera enhances viral acquisition by mosquitoes.

    PubMed

    Liu, Jianying; Liu, Yang; Nie, Kaixiao; Du, Senyan; Qiu, Jingjun; Pang, Xiaojing; Wang, Penghua; Cheng, Gong

    2016-01-01

    The arbovirus life cycle involves viral transfer between a vertebrate host and an arthropod vector, and acquisition of virus from an infected mammalian host by a vector is an essential step in this process. Here, we report that flavivirus nonstructural protein-1 (NS1), which is abundantly secreted into the serum of an infected host, plays a critical role in flavivirus acquisition by mosquitoes. The presence of dengue virus (DENV) and Japanese encephalitis virus NS1s in the blood of infected interferon-α and γ receptor-deficient mice (AG6) facilitated virus acquisition by their native mosquito vectors because the protein enabled the virus to overcome the immune barrier of the mosquito midgut. Active immunization of AG6 mice with a modified DENV NS1 reduced DENV acquisition by mosquitoes and protected mice against a lethal DENV challenge, suggesting that immunization with NS1 could reduce the number of virus-carrying mosquitoes as well as the incidence of flaviviral diseases. Our study demonstrates that flaviviruses utilize NS1 proteins produced during their vertebrate phases to enhance their acquisition by vectors, which might be a result of flavivirus evolution to adapt to multiple host environments. PMID:27562253

  7. Identification of Damaging nsSNVs in HumanERCC2 Gene.

    PubMed

    Fang, Shuo; Zhang, Yuntong; Xu, Miao; Xue, Chunyu; He, Lin; Cai, Lei; Xing, Xin

    2016-09-01

    The hERCC2 gene is an important DNA repair molecule for initiating Cutaneous melanoma (CM). Therefore, it is advisable to study the possible functional SNVs in hERCC2. To achieve this goal, we collected total 2, 253 SNVs in hERCC2from the EMBL website, of which 303 are non-synonymous single nucleotide variants (nsSNVs). Then, SIFT and PolyPhen were used to predict the damaging nsSNVs, and four nsSNVs (rs368866996, rs377739017, rs370819591, and rs121913022) were suggested to be damaging mutations. Since I-Mutant2.0 showed a decrease in stability for the mutants containing each of the four nsSNVs, a 3D protein structure was modeled. Based on the comparison of the energy after minimization, RMSD and stabilizing residues between the native and mutant proteins' structure, rs121913022 was proposed to be the most damaging variant among the nsSNVs in hERCC2 gene by decreasing the stability of protein. The mutant G713R of hERCC2 protein caused by rs121913022 was found to have less expression level than native hERCC2 protein in melanoma cells. These results suggest that rs121913022 may have potentially important clinical and drug target implications.

  8. In-Silico Computing of the Most Deleterious nsSNPs in HBA1 Gene

    PubMed Central

    AbdulAzeez, Sayed; Borgio, J. Francis

    2016-01-01

    Background α-Thalassemia (α-thal) is a genetic disorder caused by the substitution of single amino acid or large deletions in the HBA1 and/or HBA2 genes. Method Using modern bioinformatics tools as a systematic in-silico approach to predict the deleterious SNPs in the HBA1 gene and its significant pathogenic impact on the functions and structure of HBA1 protein was predicted. Results and Discussion A total of 389 SNPs in HBA1 were retrieved from dbSNP database, which includes: 201 non-coding synonymous (nsSNPs), 43 human active SNPs, 16 intronic SNPs, 11 mRNA 3′ UTR SNPs, 9 coding synonymous SNPs, 9 5′ UTR SNPs and other types. Structural homology-based method (PolyPhen) and sequence homology-based tool (SIFT), SNPs&Go, PROVEAN and PANTHER revealed that 2.4% of the nsSNPs are pathogenic. Conclusions A total of 5 nsSNPs (G60V, K17M, K17T, L92F and W15R) were predicted to be responsible for the structural and functional modifications of HBA1 protein. It is evident from the deep comprehensive in-silico analysis that, two nsSNPs such as G60Vand W15R in HBA1 are highly deleterious. These “2 pathogenic nsSNPs” can be considered for wet-lab confirmatory analysis. PMID:26824843

  9. Analysis of the PDZ binding specificities of Influenza A Virus NS1 proteins

    PubMed Central

    2011-01-01

    The Influenza A virus non-structural protein 1 (NS1) is a multifunctional virulence factor with several protein-protein interaction domains, involved in preventing apoptosis of the infected cell and in evading the interferon response. In addition, the majority of influenza A virus NS1 proteins have a class I PDZ-binding motif at the C-terminus, and this itself has been shown to be a virulence determinant. In the majority of human influenza NS1 proteins the consensus motif is RSxV: in avian NS1 it is ESxV. Of the few human strains that have the avian motif, all were from very high mortality outbreaks of the disease. Previous work has shown that minor differences in PDZ-binding motifs can have major effects on the spectrum of cellular proteins targeted. In this study we analyse the effect of these differences upon the binding of Influenza A virus NS1 protein to a range of cellular proteins involved in polarity and signal transduction. PMID:21247458

  10. Topography of the simian rotavirus nonstructural glycoprotein (NS28) in the endoplasmic reticulum membrane.

    PubMed

    Chan, W K; Au, K S; Estes, M K

    1988-06-01

    The simian rotavirus SA11 genome segment 10 codes for a nonstructural glycoprotein, NS28, that has been hypothesized to be involved in budding of viral particles into the endoplasmic reticulum (ER) membrane. Previous studies had suggested that NS28 is an integral membrane protein of the ER, possibly a transmembrane protein. We have examined the topography of NS28 inserted in microsomal membranes following cell-free translation of genome segment 10 transcripts. These transcripts were obtained either by hybrid selection of mRNA synthesized by the endogenous viral RNA polymerase or by in vitro transcription of genome segment 10 cDNA using SP6 polymerase. Full-length and truncated gene 10 transcripts were translated in a cell-free system supplemented with dog pancreatic microsomes. The existence of a cytoplasmic domain of the translation product was demonstrated by protease protection experiments. An 18,000 (18K) mol wt glycosylated polypeptide was protected from digestion with proteinase K and trypsin, whereas chymotrypsin digestion yielded a 23K mol wt glycosylated polypeptide. Correlation of these biochemical data with the known sequence of NS28 suggests that a 10K mol wt hydrophilic, carboxy-terminal fragment (from amino acid number 86 to amino acid number 175) of this glycoprotein is exposed on the cytoplasmic side of the ER membrane. A model of how NS28 folds in the ER membrane is proposed. PMID:2835861

  11. Characterization of methicillin-resistant coagulase-negative staphylococci (MRCoNS) in retail meat.

    PubMed

    Bhargava, Kanika; Zhang, Yifan

    2014-09-01

    This study was to understand the extent of methicillin-resistant coagulase-negative staphylococci (MRCoNS) serving as a mecA reservoir in retail meat. MRCoNS were isolated from retail meat (beef, chicken, and turkey) in Detroit and characterized by sodA gene sequencing for species identification, staphylococcal cassette chromosome mec (SCCmec) typing, and pulsed-field gel electrophoresis (PFGE). Unique MRCoNS isolates recovered from 25 meat samples were comprised of Staphylococcus sciuri (n = 13), Staphylococcus fleuretti (n = 4), Staphylococcus lentus (n = 3), Staphylococcus epidermidis (n = 2), Staphylococcus vitulinus (n = 1), Staphylococcus saprophyticus (n = 1) and Staphylococcus pasteuri (n = 1). Heterogeneous and composite SCCmec types, including I, III, IV, V, I + V and III + V were identified in 16 isolates. Same SCCmec types were recovered in different staphylococcal species and meat sources. Indistinguishable PFGE patterns were also observed in S. sciuri isolated from beef, chicken, and turkey, and with different SCCmec types. In conclusion, multiple CoNS species can serve as reservoirs for mecA. In addition to the clonal transmission of MRCoNS in meat, horizontal occurrence of SCCmec is observed in staphylococcal species. PMID:24929717

  12. Structure-guided Discovery of a Novel Non-peptide Inhibitor of Dengue Virus NS2B-NS3 Protease.

    PubMed

    Li, Linfeng; Basavannacharya, Chandrakala; Chan, Kitti Wing Ki; Shang, Luqing; Vasudevan, Subhash G; Yin, Zheng

    2015-09-01

    Dengue fever is a fast emerging epidemic-prone viral disease caused by dengue virus serotypes 1-4. NS2B-NS3 protease of dengue virus is a validated target to develop antiviral agents. A major limitation in developing dengue virus protease inhibitors has been the lack of or poor cellular activity. In this work, we extracted and refined a pharmacophore model based on X-ray crystal structure and predicted binding patterns, followed by a three-dimensional flexible database filtration. These output molecules were screened according to a docking-based protocol, leading to the discovery of a compound with novel scaffold and good cell-based bioactivity that has potential to be further optimized. The discovery of this novel scaffold by combination of in silico methods suggests that structure-guided drug discovery can lead to the development of potent dengue virus protease inhibitors.

  13. Mutations within the conserved NS1 nuclear export signal lead to inhibition of influenza A virus replication

    PubMed Central

    2014-01-01

    Background The influenza A virus NS1 protein is a virulence factor and an antagonist of host cell innate immune responses. During virus infection NS1 protein has several functions both in the nucleus and in the cytoplasm and its intracellular localization is regulated by one or two nuclear localization signals (NLS) and a nuclear export signal (NES). Methods In order to investigate the role of NS1 NES in intracellular localization, virus life cycle and host interferon responses, we generated recombinant A/Udorn/72 viruses harboring point mutations in the NES sequence. Results NS1 NES was found to be inactivated by several of the mutations resulting in nuclear retention of NS1 at late stages of infection confirming that this sequence is a bona fide functional NES. Some of the mutant viruses showed reduced growth properties in cell culture, inability to antagonize host cell interferon production and increased p-IRF3 levels, but no clear correlation between these phenotypes and NS1 localization could be made. Impaired activation of Akt phosphorylation by the replication-deficient viruses indicates possible disruption of NS1-p85β interaction by mutations in the NES region. Conclusion We conclude that mutations within the NS1 NES result in impairment of several NS1 functions which extends further from the NES site being only involved in regulating the nuclear-cytoplasmic trafficking of NS1. PMID:25023993

  14. Structural insight and flexible features of NS5 proteins from all four serotypes of Dengue virus in solution.

    PubMed

    Saw, Wuan Geok; Tria, Giancarlo; Grüber, Ardina; Subramanian Manimekalai, Malathy Sony; Zhao, Yongqian; Chandramohan, Arun; Srinivasan Anand, Ganesh; Matsui, Tsutomu; Weiss, Thomas M; Vasudevan, Subhash G; Grüber, Gerhard

    2015-11-01

    Infection by the four serotypes of Dengue virus (DENV-1 to DENV-4) causes an important arthropod-borne viral disease in humans. The multifunctional DENV nonstructural protein 5 (NS5) is essential for capping and replication of the viral RNA and harbours a methyltransferase (MTase) domain and an RNA-dependent RNA polymerase (RdRp) domain. In this study, insights into the overall structure and flexibility of the entire NS5 of all four Dengue virus serotypes in solution are presented for the first time. The solution models derived revealed an arrangement of the full-length NS5 (NS5FL) proteins with the MTase domain positioned at the top of the RdRP domain. The DENV-1 to DENV-4 NS5 forms are elongated and flexible in solution, with DENV-4 NS5 being more compact relative to NS5 from DENV-1, DENV-2 and DENV-3. Solution studies of the individual MTase and RdRp domains show the compactness of the RdRp domain as well as the contribution of the MTase domain and the ten-residue linker region to the flexibility of the entire NS5. Swapping the ten-residue linker between DENV-4 NS5FL and DENV-3 NS5FL demonstrated its importance in MTase-RdRp communication and in concerted interaction with viral and host proteins, as probed by amide hydrogen/deuterium mass spectrometry. Conformational alterations owing to RNA binding are presented.

  15. The methyltransferase domain of dengue virus protein NS5 ensures efficient RNA synthesis initiation and elongation by the polymerase domain.

    PubMed

    Potisopon, Supanee; Priet, Stéphane; Collet, Axelle; Decroly, Etienne; Canard, Bruno; Selisko, Barbara

    2014-10-01

    Viral RNA-dependent RNA polymerases (RdRps) responsible for the replication of single-strand RNA virus genomes exert their function in the context of complex replication machineries. Within these replication complexes the polymerase activity is often highly regulated by RNA elements, proteins or other domains of multi-domain polymerases. Here, we present data of the influence of the methyltransferase domain (NS5-MTase) of dengue virus (DENV) protein NS5 on the RdRp activity of the polymerase domain (NS5-Pol). The steady-state polymerase activities of DENV-2 recombinant NS5 and NS5-Pol are compared using different biochemical assays allowing the dissection of the de novo initiation, transition and elongation steps of RNA synthesis. We show that NS5-MTase ensures efficient RdRp activity by stimulating the de novo initiation and the elongation phase. This stimulation is related to a higher affinity of NS5 toward the single-strand RNA template indicating NS5-MTase either completes a high-affinity RNA binding site and/or promotes the correct formation of the template tunnel. Furthermore, the NS5-MTase increases the affinity of the priming nucleotide ATP upon de novo initiation and causes a higher catalytic efficiency of the polymerase upon elongation. The complex stimulation pattern is discussed under the perspective that NS5 adopts several conformations during RNA synthesis. PMID:25209234

  16. The methyltransferase domain of dengue virus protein NS5 ensures efficient RNA synthesis initiation and elongation by the polymerase domain

    PubMed Central

    Potisopon, Supanee; Priet, Stéphane; Collet, Axelle; Decroly, Etienne; Canard, Bruno; Selisko, Barbara

    2014-01-01

    Viral RNA-dependent RNA polymerases (RdRps) responsible for the replication of single-strand RNA virus genomes exert their function in the context of complex replication machineries. Within these replication complexes the polymerase activity is often highly regulated by RNA elements, proteins or other domains of multi-domain polymerases. Here, we present data of the influence of the methyltransferase domain (NS5-MTase) of dengue virus (DENV) protein NS5 on the RdRp activity of the polymerase domain (NS5-Pol). The steady-state polymerase activities of DENV-2 recombinant NS5 and NS5-Pol are compared using different biochemical assays allowing the dissection of the de novo initiation, transition and elongation steps of RNA synthesis. We show that NS5-MTase ensures efficient RdRp activity by stimulating the de novo initiation and the elongation phase. This stimulation is related to a higher affinity of NS5 toward the single-strand RNA template indicating NS5-MTase either completes a high-affinity RNA binding site and/or promotes the correct formation of the template tunnel. Furthermore, the NS5-MTase increases the affinity of the priming nucleotide ATP upon de novo initiation and causes a higher catalytic efficiency of the polymerase upon elongation. The complex stimulation pattern is discussed under the perspective that NS5 adopts several conformations during RNA synthesis. PMID:25209234

  17. Hepatitis C virus nonstructural protein NS3 transforms NIH 3T3 cells.

    PubMed Central

    Sakamuro, D; Furukawa, T; Takegami, T

    1995-01-01

    Clinical evidence suggests that hepatitis C virus (HCV) is etiologically involved in hepatic cancer and liver cirrhosis. To investigate whether the HCV nonstructural protein NS3 has oncogenic activity, NIH 3T3 cells were transfected with an expression vector containing cDNA for the 5'- or 3'-half sequence of the HCV genome segment encoding NS3. Only cells transfected with the 5'-half cDNA rapidly proliferated, lost contact inhibition, grew anchorage independently in soft agar, and formed tumors in nude mice. PCR analysis confirmed the presence of the 5'-half DNA in the transfectants. These results suggest that the 5' region of the HCV genome segment encoding NS3 is involved in cell transformation. PMID:7745741

  18. Antiviral phytochemicals identification from Azadirachta indica leaves against HCV NS3 protease: an in silico approach.

    PubMed

    Ashfaq, Usman Ali; Jalil, Asma; Ul Qamar, Muhammad Tahir

    2016-08-01

    Hepatitis C virus (HCV) is a major health problem across the world affecting the people of all age groups. It is the main cause of hepatitis and at chronic stage causes liver cirrhosis and hepatocellular carcinoma. Various therapeutics are made against HCV but still there is a need to find out potential therapeutics to combat the virus. The goal of this study is to identify the phytochemicals of Azadirachta indica leaves having antiviral activity against HCV NS3 protease through molecular docking and simulation approach. Results show that the compound 3-Deacetyl-3-cinnamoyl-azadirachtin possesses good binding properties with HCV NS3/4A protease. It can be concluded from this study that Deacetyl-3-cinnamoyl-azadirachtin may serve as a potential inhibitor against NS3/4A protease. PMID:26274064

  19. Producing human ceramide-NS by metabolic engineering using yeast Saccharomyces cerevisiae.

    PubMed

    Murakami, Suguru; Shimamoto, Toshi; Nagano, Hideaki; Tsuruno, Masahiro; Okuhara, Hiroaki; Hatanaka, Haruyo; Tojo, Hiromasa; Kodama, Yukiko; Funato, Kouichi

    2015-11-17

    Ceramide is one of the most important intercellular components responsible for the barrier and moisture retention functions of the skin. Because of the risks involved with using products of animal origin and the low productivity of plants, the availability of ceramides is currently limited. In this study, we successfully developed a system that produces sphingosine-containing human ceramide-NS in the yeast Saccharomyces cerevisiae by eliminating the genes for yeast sphingolipid hydroxylases (encoded by SUR2 and SCS7) and introducing the gene for a human sphingolipid desaturase (encoded by DES1). The inactivation of the ceramidase gene YDC1, overexpression of the inositol phosphosphingolipid phospholipase C gene ISC1, and endoplasmic reticulum localization of the DES1 gene product resulted in enhanced production of ceramide-NS. The engineered yeast strains can serve as hosts not only for providing a sustainable source of ceramide-NS but also for developing further systems to produce sphingosine-containing sphingolipids.

  20. TRIM52 inhibits Japanese Encephalitis Virus replication by degrading the viral NS2A

    PubMed Central

    Fan, Wenchun; Wu, Mengge; Qian, Suhong; Zhou, Yun; Chen, Huanchun; Li, Xiangmin; Qian, Ping

    2016-01-01

    The members of tripartite-motif containing (TRIM) protein participate in various cellular processes and play an important role in host antiviral function. TRIM proteins exert their antiviral activity either directly by degrading viral proteins through their E3 ligase activity, or indirectly by promoting host innate immunity. This study demonstrated for the first time that TRIM52 is a novel antiviral TRIM protein against Japanese encephalitis virus (JEV) infection. Overexpression of TRIM52 restricted JEV replication in BHK-21 and 293T cells. In addition, JEV nonstructural protein 2A (NS2A) is a protein that interacts with TRIM52. Their interaction degraded NS2A in a proteasome-dependent manner via the E3 ligase activity of TRIM52. Thus, TRIM52 is a novel antiviral TRIM protein, and it exerted antiviral activity against JEV infection by targeting and degrading viral NS2A. PMID:27667714

  1. H-NS Regulates Gene Expression and Compacts the Nucleoid: Insights from Single-Molecule Experiments

    PubMed Central

    Winardhi, Ricksen S.; Yan, Jie; Kenney, Linda J.

    2015-01-01

    A set of abundant nucleoid-associated proteins (NAPs) play key functions in organizing the bacterial chromosome and regulating gene transcription globally. Histone-like nucleoid structuring protein (H-NS) is representative of a family of NAPs that are widespread across bacterial species. They have drawn extensive attention due to their crucial function in gene silencing in bacterial pathogens. Recent rapid progress in single-molecule manipulation and imaging technologies has made it possible to directly probe DNA binding by H-NS, its impact on DNA conformation and topology, and its competition with other DNA-binding proteins at the single-DNA-molecule level. Here, we review recent findings from such studies, and provide our views on how these findings yield new insights into the understanding of the roles of H-NS family members in DNA organization and gene silencing. PMID:26445432

  2. Hepatitis C virus NS2 protein activates cellular cyclic AMP-dependent pathways

    SciTech Connect

    Kim, Kyoung Mi; Kwon, Shi-Nae; Kang, Ju-Il; Lee, Song Hee; Jang, Sung Key; Ahn, Byung-Yoon; Kim, Yoon Ki . E-mail: yk-kim@korea.ac.kr

    2007-05-18

    Chronic infection of the hepatitis C virus (HCV) leads to liver cirrhosis and cancer. The mechanism leading to viral persistence and hepatocellular carcinoma, however, has not been fully understood. In this study, we show that the HCV infection activates cellular cAMP-dependent pathways. Expression of a luciferase reporter gene controlled by a basic promoter with the cAMP response element (CRE) was significantly elevated in human hepatoma Huh-7 cells infected with the HCV JFH1. Analysis with viral subgenomic replicons indicated that the HCV NS2 protein is responsible for the effect. Furthermore, the level of cellular transcripts whose stability is known to be regulated by cAMP was specifically reduced in cells harboring NS2-expressing replicons. These results allude to the HCV NS2 protein having a novel function of regulating cellular gene expression and proliferation through the cAMP-dependent pathway.

  3. Potent inhibitors of HCV-NS3 protease derived from boronic acids

    SciTech Connect

    Venkatraman, Srikanth; Wu, Wanli; Prongay, Andrew; Girijavallabhan, Viyyoor; Njoroge, F. George

    2009-07-23

    Chronic hepatitis C infection is the leading causes for cirrhosis of the liver and hepatocellular carcinoma, leading to liver failure and liver transplantation. The etiological agent, HCV virus produces a single positive strand of RNA that is processed with the help of serine protease NS3 to produce mature virus. Inhibition of NS3 protease can be potentially used to develop effective drugs for HCV infections. Numerous efforts are now underway to develop potent inhibitors of HCV protease that contain ketoamides as serine traps. Herein we report the synthesis of a series of potent inhibitors that contain a boronic acid as a serine trap. The activity of these compounds were optimized to 200 pM. X-ray structure of compound 17 bound to NS3 protease is also discussed.

  4. Producing human ceramide-NS by metabolic engineering using yeast Saccharomyces cerevisiae.

    PubMed

    Murakami, Suguru; Shimamoto, Toshi; Nagano, Hideaki; Tsuruno, Masahiro; Okuhara, Hiroaki; Hatanaka, Haruyo; Tojo, Hiromasa; Kodama, Yukiko; Funato, Kouichi

    2015-01-01

    Ceramide is one of the most important intercellular components responsible for the barrier and moisture retention functions of the skin. Because of the risks involved with using products of animal origin and the low productivity of plants, the availability of ceramides is currently limited. In this study, we successfully developed a system that produces sphingosine-containing human ceramide-NS in the yeast Saccharomyces cerevisiae by eliminating the genes for yeast sphingolipid hydroxylases (encoded by SUR2 and SCS7) and introducing the gene for a human sphingolipid desaturase (encoded by DES1). The inactivation of the ceramidase gene YDC1, overexpression of the inositol phosphosphingolipid phospholipase C gene ISC1, and endoplasmic reticulum localization of the DES1 gene product resulted in enhanced production of ceramide-NS. The engineered yeast strains can serve as hosts not only for providing a sustainable source of ceramide-NS but also for developing further systems to produce sphingosine-containing sphingolipids. PMID:26573460

  5. Alkylated flavanones from the bark of Cryptocarya chartacea as dengue virus NS5 polymerase inhibitors.

    PubMed

    Allard, Pierre-Marie; Dau, Elise Tran Huu; Eydoux, Cécilia; Guillemot, Jean-Claude; Dumontet, Vincent; Poullain, Cyril; Canard, Bruno; Guéritte, Françoise; Litaudon, Marc

    2011-11-28

    An in vitro screening of New Caledonian plants allowed the selection of several species with a significant dengue virus NS5 RNA-dependent RNA polymerase (RdRp) inhibiting activity. The chemical investigation of Cryptocarya chartacea led to the isolation of a series of new mono- and dialkylated flavanones named chartaceones A-F (1-6), along with pinocembrin. They were isolated as racemic mixtures and characterized using extensive one- and two-dimensional NMR spectroscopy. Four diastereomers of chartaceone A (1) were separated using chiral HPLC, and their absolute configurations were established by comparison of their experimental and calculated ECD spectra. The dialkylated flavanones, chartaceones C-F (3-6), exhibited the most significant NS5 RdRp inhibiting activity, with IC(50) ranging from 1.8 to 4.2 μM. Chartaceones represent a new class of non-nucleosidic inhibitors of the DENV NS5 RdRp. PMID:22050318

  6. Producing human ceramide-NS by metabolic engineering using yeast Saccharomyces cerevisiae

    PubMed Central

    Murakami, Suguru; Shimamoto, Toshi; Nagano, Hideaki; Tsuruno, Masahiro; Okuhara, Hiroaki; Hatanaka, Haruyo; Tojo, Hiromasa; Kodama, Yukiko; Funato, Kouichi

    2015-01-01

    Ceramide is one of the most important intercellular components responsible for the barrier and moisture retention functions of the skin. Because of the risks involved with using products of animal origin and the low productivity of plants, the availability of ceramides is currently limited. In this study, we successfully developed a system that produces sphingosine-containing human ceramide-NS in the yeast Saccharomyces cerevisiae by eliminating the genes for yeast sphingolipid hydroxylases (encoded by SUR2 and SCS7) and introducing the gene for a human sphingolipid desaturase (encoded by DES1). The inactivation of the ceramidase gene YDC1, overexpression of the inositol phosphosphingolipid phospholipase C gene ISC1, and endoplasmic reticulum localization of the DES1 gene product resulted in enhanced production of ceramide-NS. The engineered yeast strains can serve as hosts not only for providing a sustainable source of ceramide-NS but also for developing further systems to produce sphingosine-containing sphingolipids. PMID:26573460

  7. Bright-White Beetle Scales Optimise Multiple Scattering of Light

    NASA Astrophysics Data System (ADS)

    Burresi, Matteo; Cortese, Lorenzo; Pattelli, Lorenzo; Kolle, Mathias; Vukusic, Peter; Wiersma, Diederik S.; Steiner, Ullrich; Vignolini, Silvia

    2014-08-01

    Whiteness arises from diffuse and broadband reflection of light typically achieved through optical scattering in randomly structured media. In contrast to structural colour due to coherent scattering, white appearance generally requires a relatively thick system comprising randomly positioned high refractive-index scattering centres. Here, we show that the exceptionally bright white appearance of Cyphochilus and Lepidiota stigma beetles arises from a remarkably optimised anisotropy of intra-scale chitin networks, which act as a dense scattering media. Using time-resolved measurements, we show that light propagating in the scales of the beetles undergoes pronounced multiple scattering that is associated with the lowest transport mean free path reported to date for low-refractive-index systems. Our light transport investigation unveil high level of optimisation that achieves high-brightness white in a thin low-mass-per-unit-area anisotropic disordered nanostructure.

  8. Space station contamination study: Assessment of contaminant spectral brightness

    NASA Technical Reports Server (NTRS)

    Torr, D. G.

    1988-01-01

    The assessment of spectral brightness resulting from the ambient-contaminant interaction requires a knowledge of the details of cross sections and excitation mechanisms. The approach adopted was to utilize the spectral brightness measurements made on Spacelab 1 and on the S3-4 spacecraft to identify source mechanisms, key cross sections and hence, the abundance of contaminant species. These inferred abundances were then used to update the composition comprising the total column concentrations predicted by the Science and Engineering Associates' configuration contamination model for the Space Station and to scale the irradiances to four altitudes: 300, 350, 400, and 463 km. The concentration irradiances are compared with zodiacal natural background levels. The results demonstrate that emissive contamination is significantly more severe than anticipated. It is shown that spectral emissions can become competitive with the zodiacal background up to altitudes as high as 400 km for the vacuum ultraviolet and visible emissions.

  9. Sunspot bright rings and the thermal diffusivity of solar convection

    NASA Technical Reports Server (NTRS)

    Duvall, T., Jr.; Fowler, L. A.; Foukal, P.

    1983-01-01

    Raster-scan observations of 10 sunspots, made in 1980 and 1981 with the 512-channel diode array and vacuum telescope at the Kitt Peak National Observatory, are reported. Data from several 10-min scans of 0.25-A passbands of clean continuum were summed to give an rms noise level of 0.25 percent, corrected by applying a limb-darkening curve, and analyzed to determine the average intensity for each of eight segments of a series of concentric rings around each sunspot. Faculae and pores were identified and discarded in constructing radial intensity profiles. Marginally significant bright symmetric rings (peak amplitude 0.1-0.3 percent) not attributable to residual facular signal or instrumental effects were observed around 6 of 10 sunspots. No evidence of more intense bright rings was found. These findings are discussed in terms of thermal-diffusion models proposed to explain the fate of the radiative flux blocked by sunspots.

  10. Autostereoscopic display with high brightness and power efficiency

    NASA Astrophysics Data System (ADS)

    Eichenlaub, Jesse B.

    1994-04-01

    Dimension Technologies Inc. has experimentally demonstrated an optical system that produces autostereoscopic images and also allows very high brightness and power efficiency to be achieved using off the shelf color LCDs. This capability is important in applications such as cockpit displays or mobile, portable, or laptop systems where brightness must be maximized but power conserved as much as possible. The effects are achieved through the creation of light line illumination, by means of which autostereoscopic images are produced, and by simultaneously concentrating the light emitted by the display toward the area the viewer's head is. By turning different illumination sources on and off, it is possible to aim both the concentration area and the 3D viewing area at the observer's head as the observer moves. A variation on the system allows two or more persons to be tracked independently. Cross talk (ghosting) can be reduced to the point that imperceptibility can be achieved.

  11. Night-sky brightness at observatories and sites

    NASA Astrophysics Data System (ADS)

    Garstang, R. H.

    1989-03-01

    A model previously constructed for night-sky brightness calculations has been modified to allow for the curvature of the earth. The model has been applied to calculate the brightness at the following observatories: Mount Wilson, Lick, Mount Palomar, Kitt Peak, Sacramento Peak, Mauna Kea, McDonald, San Pedro Martir, Mount Hopkins, Mount Lemmon, Lowell (Mars Hill), Lowell (Anderson Mesa), Fick, Iowa, Van Vleck, David Dunlap, Anglo-Australian, Haute Provence, and Cerro Tololo. Calculations have also been carried out for the following prospective observatory sites: Junipero Serra, Mount Graham, Charleston Peak, Wheeler Peak, Miller Peak, San Benito Mountain, Lowell (Hutch Mountain), Lowell (Saddle Mountain), and South Baldy (New Mexico). The model is extended to calculate magnitudes in the B photometric band.

  12. Bright-white beetle scales optimise multiple scattering of light.

    PubMed

    Burresi, Matteo; Cortese, Lorenzo; Pattelli, Lorenzo; Kolle, Mathias; Vukusic, Peter; Wiersma, Diederik S; Steiner, Ullrich; Vignolini, Silvia

    2014-01-01

    Whiteness arises from diffuse and broadband reflection of light typically achieved through optical scattering in randomly structured media. In contrast to structural colour due to coherent scattering, white appearance generally requires a relatively thick system comprising randomly positioned high refractive-index scattering centres. Here, we show that the exceptionally bright white appearance of Cyphochilus and Lepidiota stigma beetles arises from a remarkably optimised anisotropy of intra-scale chitin networks, which act as a dense scattering media. Using time-resolved measurements, we show that light propagating in the scales of the beetles undergoes pronounced multiple scattering that is associated with the lowest transport mean free path reported to date for low-refractive-index systems. Our light transport investigation unveil high level of optimisation that achieves high-brightness white in a thin low-mass-per-unit-area anisotropic disordered nanostructure. PMID:25123449

  13. Schmidt modes in the angular spectrum of bright squeezed vacuum

    NASA Astrophysics Data System (ADS)

    Sharapova, P.; Pérez, A. M.; Tikhonova, O. V.; Chekhova, M. V.

    2015-04-01

    We investigate both theoretically and experimentally strong correlations in macroscopic (bright) quantum states of light generated via unseeded parametric down-conversion and four-wave mixing. The states generated this way contain only quantum noise, without a classical component, and are referred to as bright squeezed vacuum (BSV). Their important advantage is the multimode structure, which offers a larger capacity for the encoding of quantum information. For the theoretical description of these states and their correlation features we introduce a generalized fully analytical approach, based on the concept of independent collective (Schmidt) modes and valid for the cases of both weak and strong nonlinear interaction. In experiment, we generate states of macroscopic BSV with up to 1010 photons per mode and examine large photon-number spatial correlations that are found to be very well described by our theoretical approach.

  14. Fast microtomography using bright monochromatic x-rays

    SciTech Connect

    Jung, J. W.; Lee, J. S.; Park, S. J.; Chang, S.; Pyo, J.; Kwon, N.; Kim, J.; Kohmura, Y.; Nishino, Y.; Yamamoto, M.; Ishikawa, T.

    2012-09-15

    A fast microtomography system for high-resolution high-speed imaging has been developed using bright monochromatic x-rays at the BL29XU beamline of SPring-8. The shortest scan time for microtomography we attained was 0.25 s in 1.25 {mu}m effective pixel size by combining the bright monochromatic x-rays, a fast rotating sample stage, and a high performance x-ray imaging detector. The feasibility of the tomography system was successfully demonstrated by visualization of rising bubbles in a viscous liquid, an interesting issue in multiphase flow physics. This system also provides a high spatial (a measurable feature size of 300 nm) or a very high temporal (9.8 {mu}s) resolution in radiographs.

  15. Operational Performance Improvements to BRIght Target Explorer Constellation

    NASA Astrophysics Data System (ADS)

    Choi, Seung Yun

    The BRIght Target Explorer (BRITE)-Constellation is composed of six nano-satellites funded by Austria, Canada, and Poland, and each of them is equipped with an optical telescope that observes stars with visual magnitude +3.5 or brighter. BRITE-Constellation has provided numerous images of bright stars from Low Earth Orbit, which will eventually lead to investigation of origin of the Universe. This thesis presents the contribution of the author to BRITE mission, especially in BRITE Operations. The author performed antenna steering experiments on UniBRITE and BRITE-Toronto, to improve data downlink. To improve scientific data collection from BRITE satellites, the author computed available observation time for multiple targets every orbit, which resulted in collection of twice the amount of scientific data. Also, the author increased the available observation time for each target from 32 minutes to 48 minutes by improving the performance of the star tracker on-board BRITE-Toronto.

  16. Bright-White Beetle Scales Optimise Multiple Scattering of Light

    PubMed Central

    Burresi, Matteo; Cortese, Lorenzo; Pattelli, Lorenzo; Kolle, Mathias; Vukusic, Peter; Wiersma, Diederik S.; Steiner, Ullrich; Vignolini, Silvia

    2014-01-01

    Whiteness arises from diffuse and broadband reflection of light typically achieved through optical scattering in randomly structured media. In contrast to structural colour due to coherent scattering, white appearance generally requires a relatively thick system comprising randomly positioned high refractive-index scattering centres. Here, we show that the exceptionally bright white appearance of Cyphochilus and Lepidiota stigma beetles arises from a remarkably optimised anisotropy of intra-scale chitin networks, which act as a dense scattering media. Using time-resolved measurements, we show that light propagating in the scales of the beetles undergoes pronounced multiple scattering that is associated with the lowest transport mean free path reported to date for low-refractive-index systems. Our light transport investigation unveil high level of optimisation that achieves high-brightness white in a thin low-mass-per-unit-area anisotropic disordered nanostructure. PMID:25123449

  17. Ultra-bright pulsed electron beam with low longitudinal emittance

    DOEpatents

    Zolotorev, Max

    2010-07-13

    A high-brightness pulsed electron source, which has the potential for many useful applications in electron microscopy, inverse photo-emission, low energy electron scattering experiments, and electron holography has been described. The source makes use of Cs atoms in an atomic beam. The source is cycled beginning with a laser pulse that excites a single Cs atom on average to a band of high-lying Rydberg nP states. The resulting valence electron Rydberg wave packet evolves in a nearly classical Kepler orbit. When the electron reaches apogee, an electric field pulse is applied that ionizes the atom and accelerates the electron away from its parent ion. The collection of electron wave packets thus generated in a series of cycles can occupy a phase volume near the quantum limit and it can possess very high brightness. Each wave packet can exhibit a considerable degree of coherence.

  18. Three Conformational Snapshots of the Hepatitis Virus NS3 Helicase Reveal a Ratchet Translocation Mechanism

    SciTech Connect

    Gu, M.; Rice, C

    2010-01-01

    A virally encoded superfamily-2 (SF2) helicase (NS3h) is essential for the replication of hepatitis C virus, a leading cause of liver disease worldwide. Efforts to elucidate the function of NS3h and to develop inhibitors against it, however, have been hampered by limited understanding of its molecular mechanism. Here we show x-ray crystal structures for a set of NS3h complexes, including ground-state and transition-state ternary complexes captured with ATP mimics (ADP {center_dot} BeF{sub 3} and ADP {center_dot} AlF{sub 4}{sup -}). These structures provide, for the first time, three conformational snapshots demonstrating the molecular basis of action for a SF2 helicase. Upon nucleotide binding, overall domain rotation along with structural transitions in motif V and the bound DNA leads to the release of one base from the substrate base-stacking row and the loss of several interactions between NS3h and the 3{prime} DNA segment. As nucleotide hydrolysis proceeds into the transition state, stretching of a 'spring' helix and another overall conformational change couples rearrangement of the (d)NTPase active site to additional hydrogen-bonding between NS3h and DNA. Together with biochemistry, these results demonstrate a 'ratchet' mechanism involved in the unidirectional translocation and define the step size of NS3h as one base per nucleotide hydrolysis cycle. These findings suggest feasible strategies for developing specific inhibitors to block the action of this attractive, yet largely unexplored drug target.

  19. Generation of replication-proficient influenza virus NS1 point mutants with interferon-hyperinducer phenotype.

    PubMed

    Pérez-Cidoncha, Maite; Killip, Marian J; Asensio, Víctor J; Fernández, Yolanda; Bengoechea, José A; Randall, Richard E; Ortín, Juan

    2014-01-01

    The NS1 protein of influenza A viruses is the dedicated viral interferon (IFN)-antagonist. Viruses lacking NS1 protein expression cannot multiply in normal cells but are viable in cells deficient in their ability to produce or respond to IFN. Here we report an unbiased mutagenesis approach to identify positions in the influenza A NS1 protein that modulate the IFN response upon infection. A random library of virus ribonucleoproteins containing circa 40 000 point mutants in NS1 were transferred to infectious virus and amplified in MDCK cells unable to respond to interferon. Viruses that activated the interferon (IFN) response were subsequently selected by their ability to induce expression of green-fluorescent protein (GFP) following infection of A549 cells bearing an IFN promoter-dependent GFP gene. Using this approach we isolated individual mutant viruses that replicate to high titers in IFN-compromised cells but, compared to wild type viruses, induced higher levels of IFN in IFN-competent cells and had a reduced capacity to counteract exogenous IFN. Most of these viruses contained not previously reported NS1 mutations within either the RNA-binding domain, the effector domain or the linker region between them. These results indicate that subtle alterations in NS1 can reduce its effectiveness as an IFN antagonist without affecting the intrinsic capacity of the virus to multiply. The general approach reported here may facilitate the generation of replication-proficient, IFN-inducing virus mutants, that potentially could be developed as attenuated vaccines against a variety of viruses.

  20. Generation of Replication-Proficient Influenza Virus NS1 Point Mutants with Interferon-Hyperinducer Phenotype

    PubMed Central

    Pérez-Cidoncha, Maite; Killip, Marian J.; Asensio, Víctor J.; Fernández, Yolanda; Bengoechea, José A.; Randall, Richard E.; Ortín, Juan

    2014-01-01

    The NS1 protein of influenza A viruses is the dedicated viral interferon (IFN)-antagonist. Viruses lacking NS1 protein expression cannot multiply in normal cells but are viable in cells deficient in their ability to produce or respond to IFN. Here we report an unbiased mutagenesis approach to identify positions in the influenza A NS1 protein that modulate the IFN response upon infection. A random library of virus ribonucleoproteins containing circa 40 000 point mutants in NS1 were transferred to infectious virus and amplified in MDCK cells unable to respond to interferon. Viruses that activated the interferon (IFN) response were subsequently selected by their ability to induce expression of green-fluorescent protein (GFP) following infection of A549 cells bearing an IFN promoter-dependent GFP gene. Using this approach we isolated individual mutant viruses that replicate to high titers in IFN-compromised cells but, compared to wild type viruses, induced higher levels of IFN in IFN-competent cells and had a reduced capacity to counteract exogenous IFN. Most of these viruses contained not previously reported NS1 mutations within either the RNA-binding domain, the effector domain or the linker region between them. These results indicate that subtle alterations in NS1 can reduce its effectiveness as an IFN antagonist without affecting the intrinsic capacity of the virus to multiply. The general approach reported here may facilitate the generation of replication-proficient, IFN-inducing virus mutants, that potentially could be developed as attenuated vaccines against a variety of viruses. PMID:24887174