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48 records · Page 3

Tubulinlike protein from Spirochaeta bajacaliforniensis

Tubulin proteins are the fundamental subunits of all polymeric microtubule-based eukaryotic structures. Long, hollow structures each composed of 13 protofilaments as revealed by electron microscopy, microtubules (240 angstroms in diameter) are nearly ubiquitous in eukaryotes. These proteins have been the subject of intense biochemical and biophyiscal interest since the early 1970s and are of evolutionary interest as well. If tubulin-based structures (i.e., neurotubules, mitotic spindle tubules, centrioles, kinetosomes, axonemes, etc.) evolved from spirochetes by way of motility symbioses, tubulin homologies with spirochete proteins should be detectable. Tubulin proteins are widely thought to be limited to eukaryotes. Yet both azotobacters and spirochetes have shown immunological cross-reactivity with antitubulin antibodies. In neither of these studies was tubulin isolated nor any specific antigen identified as responsible for the immunoreactivity. Furthermore, although far less uniform in structure than eukaryotic microtubules, various cytoplasmic fibers and tubules (as seen by electron microscopy) have been reported in several types of prokaryotes (e.g., Spirochaeta; large termite spirochetes; treponemes; cyanobacteria; and Azotobacter. This work forms a part of our long-range study of the possible prokaryotic origin of tubulin and microtubules. Spirochetes are helically shaped gram-negative motile prokaryotes. They differ from all other bacterial in that the position of their flagella is periplasmic: their flagella lie between the inner and outer membranes of the gram-negative cell wall. Some of the largest spirochetes have longitudinally aligned 240 angstrom microtubules. Unfortunately, in spite of many attempts, all of the larger spirochetes (family Pillotaceae) with well-defined cytoplasmic tubules and antitubulin immunoreactivity are not cultivable. However, a newly described spirochete species (Spirochaeta bajacaliforniensis) possessing cytoplasmic fibers displays antitubulin immunoreactivity in whole-cell preparations. Since preliminary observations suggested that Spirochaeta bajacaliforniensis proteins may be related to eukaryotic tubulins, their characterization was undertaken. Brain tubulin can be purified by utilizing its ability to polymerize at warm temperatures and to depolymerize in the cold. After several cycles of sedimentation and redissolution the microtubule fraction is comprised of 75% tubulin and 20% high molecular mass microtubule-associated proteins (MAPs). In this paper we report that components of cell lysates, prepared from a spirochete that contains cytoplasmic fibers (Spirochaeta bajacaliforniensis), also exhibit the property of temperature-dependent cyclical sedimentation. Additionally we report the identification and characterization of the polypeptide responsible for cross-reactivity with antitubulin antiserum.

Non-NASA Center↗

Localization of the ANG II type 2 receptor in the microcirculation of skeletal muscle

Only functional studies have suggested the presence of the ANG II type 2 (AT2) receptor in the microcirculation. To determine the distribution of this receptor in the rat skeletal muscle microcirculation, a polyclonal rabbit anti-rat antiserum was developed and used for immunohistochemistry and Western blot analysis. The antiserum was prepared against a highly specific and antigenic AT2-receptor synthetic peptide and was validated by competition and sensitivity assays. Western blot analysis demonstrated a prominent, single band at approximately 40 kDa in cremaster and soleus muscle. Immunohistochemical analysis revealed a wide distribution of AT2 receptors throughout the skeletal muscle microcirculation in large and small microvessels. Microanatomic studies displayed an endothelial localization of the AT2 receptor, whereas dual labeling with smooth muscle alpha-actin also showed colocalization of the AT2 receptor with vascular smooth muscle cells. Other cells associated with the microvessels also stained positive for AT2 receptors. Briefly, this study confirms previous functional data and localizes the AT2 receptor to the microcirculation. These studies demonstrate that the AT2 receptor is present on a variety of vascular cell types and that it is situated in a fashion that would allow it to directly oppose ANG II type 1 receptor actions.

NASA Discipline Cardiopulmonary↗

Post-embedding tem signal-to-noise ratio of S-100

We assessed the reactivity of purified S-100 antiserum in immuno-electron microscopy by counting the number of gold particles per microns 2 over inner ear tissues embedded in different media. Sections containing predominantly Schwann's cell cytoplasm and nucleus, afferent fiber axoplasm and myelin sheath of chick cochleae were reacted with anti-S-100 IgG, an antibody to a calcium binding protein of neuronal tissues, then labeled with anti-IgG-gold conjugate. This investigation was conducted because previously published procedures, unmodified, did not yield acceptable results. Preparation of all specimens was identical. Only the medium (PolyBed 812, Araldite or Spurr epoxies; and LR White, LR Gold or Lowicryl plastics) was changed. The medium was made the changing variable because antigens available in post-embedding immuno-electron microscopy are decreased by heat, either used and/or released during polymerization of the embedding medium. The results indicate that: (a) none of the embedding media above provided optimal signal-to-noise ratio for all parts of the nerve stained in the same section; (b) aggregation of gold particles over cells was highest in embedding media with high background labeling over areas devoid of tissue (noise); (c) aggregation occurred randomly throughout both cellular and acellular regions; and (d) particles aggregated less and were distributed more evenly in tissues from media yielding good ultrastructural integrity.

Non-NASA Center↗

Catching Up on State Transitions in Cygnus X-1

In 2005 February we observed Cygnus X-1 over a period of 10 days quasi-continuously with the Rossi X-ray Timing Explorer and the Ryle telescope. We present the results of the spectral and timing analysis on a timescale of 90 min and show that the behavior of Cyg X-1 is similar to that found during our years long monitoring campaign. As a highlight we present evidence for a full transition from the hard to the soft state that happened during less than three hours. The observation provided a more complete picture of a state transition than before, especially concerning the evolution of the time lags, due to unique transition coverage and analysis with high time resolution.

Boeck, Moritz↗

Modelling a Simultaneous Radio/X-Ray Flare from Cyg X-1

The long-term monitoring campaign of Cyg X-1 has provided the detection of the first simultaneous radio/X-ray flare seen from that source. We investigate the physical characteristics of the event in terms of emission from a homogeneous, expanding blob of pair-plasma, superimposed on a baseline flux in both bands. We find that while the radio flare can be reconstructed under various configurations of a cooling blob, continuous (re)acceleration of particles inside the jet is necessary to sustain X-ray emission at the levels implied by the data, for the observed duration. We present major results of the modelling and discuss implications for the role of microquasar jets.

Leventis, Konstantinos↗

Probing the Accretion Disk and Central Engine Structure of the NGC 4258 with Suzaku and XMM-Newton Observations

We present an X-ray study of the low-luminosity active galactic nucleus (AGN) in NGC 4258 using data from Suzaku, XMM-Newton, and the Swift/Burst Alert Telescope survey. We find that signatures of X-ray reprocessing by cold gas are very weak in the spectrum of this Seyfert-2 galaxy; a weak, narrow fluorescent K(alpha) emission line of cod iron is robustly detected in both the Suzaku and XMM-Newton spectra but at a level much below that of most other Seyfert-2 galaxies. We conclude that the circumnuclear environment of this AGN is very "clean" and lacks the Compton-thick obscuring torus of unified Seyfert schemes. From the narrowness of the iron line, together with evidence of line flux variability between the Suzaku and XMM-Newton observations, we constrain the line emitting region to be between 3 x 10(exp 3)r(sub g) and 4 x 10(exp 4)r(sub g), from the black hole. We show that the observed properties of the iron line can be explained if the line originates from the surface layers of a warped accretion disk. In particular, we present explicit calculations of the expected iron line from a disk warped by Lens-Thirring precession from a misaligned central black hole. Finally, the Suzaku data reveal clear evidence of large amplitude 2-10 keV variability on timescales of 50 ksec and smaller amplitude flares on timescales as short as 5-10 ksec. If associated with accretion disk processes, such rapid variability requires an origin in the innermost regions of the disk (r approx. equals 10(r(sub g) or less). Analysis of the difference spectrum between a high- and low-flux states suggests that the variable component of the X-ray emission is steeper and more absorbed than the average AGN emission, suggesting that the primary X-ray source and absorbing screen have a spatial structure on comparable scales. We note the remarkable similarity between the circumnuclear environment of NGC 4258 and another well studied low-luminosity AGN, M81*.

Reynolds, Christopher S.↗

Broadband X-Ray Spectra of GX 339-4 and the Geometry of Accreting Black Holes in the Hard State

A major question in the study of black hole binaries involves our understanding of the accretion geometry when the sources are in the "hard" state, with an X-ray energy spectrum dominated by a hard power-law component and radio emission coming from a steady "compact" jet. Although the common hard state picture is that the accretion disk is truncated, perhaps at hundreds of gravitational radii (Rg) from the black hole, recent results for the recurrent transient GX 339-4 by Miller and coworkers show evidence for disk material very close to the black hole's innermost stable circular orbit. That work studied GX 339-4 at a luminosity of approximately 5% of the Eddington limit (L(sub Edd) and used parameters from a relativistic reflection model and the presence of a thermal component as diagnostics. Here we use similar diagnostics but extend the study to lower luminosities (2.3% and 0.8% L(sub Edd)) using Swift and RXTE observations of GX 339-4. We detect a thermal component with an inner disk temperature of approximately 0.2 keV at 2.3% L (sub Edd). At both luminosities, we detect broad features due to iron K-alpha that are likely related to reflection of hard X-rays off disk material. If these features are broadened by relativistic effects, they indicate that the material resides within 10 Rg, and the measurements are consistent with the disk's inner radius remaining at approximately 4 Rg down to 0.8% L(sub Edd). However, we also discuss an alternative model for the broadening, and we note that the evolution of the thermal component is not entirely consistent with the constant inner radius interpretation. Finally, we discuss the results in terms of recent theoretical work by Liu and co-workers on the possibility that material may condense out of an Advection-Dominated Accretion Flow to maintain an inner optically thick disk.

Tomsick, John A.↗

Corona, Jet, and Relativistic Line Models for Suzaku/RXTE/Chandra-HETG Observations of the Cygnus X-1 Hard State

Using Suzaku and the Rossi X-ray Timing Explorer (RXTE), we have conducted a series of four simultaneous observations of the galactic black hole candidate Cyg X-1 in what were historically faint and spectrally hard "low states". Additionally, all of these observations occurred near superior conjunction with our line of sight to the X-ray source passing through the dense phases of the "focused wind" from the mass donating secondary. One of our observations was also simultaneous with observations by the Chandra-High Energy Transmission Grating (HETG). These latter spectra are crucial for revealing the ionized absorption due to the secondary s focused wind. Such absorption is present and must be accounted for in all four spectra. These simultaneous data give an unprecedented view of the 0.8-300 keV spectrum of Cyg X-1, and hence bear upon both corona and X-ray emitting jet models of black hole hard states. Three models fit the spectra well: coronae with thermal or mixed thermal/non-thermal electron populations, and jets. All three models require a soft component that we fit with a low temperature disk spectrum with an inner radius of only a few tens of GM/c2. All three models also agree that the known spectral break at 10 keV is not solely due to the presence of reflection, but each gives a different underlying explanation for the augmentation of this break. Thus whereas all three models require that there is a relativistically broadened Fe line, the strength and inner radius of such a line is dependent upon the specific model, thus making premature line-based estimates of the black hole spin in the Cyg X-1 system. We look at the relativistic line in detail, accounting for the narrow Fe emission and ionized absorption detected by HETG. Although the specific relativistic parameters of the line are continuum-dependent, none of the broad line fits allow for an inner disk radius that is > 40 GM/c(sup 2).

Nowak, Michael A.↗

The Event Horizon General Relativistic Magnetohydrodynamic Code Comparison Project

Recent developments in compact object astrophysics, especially the discovery of merging neutron stars by LIGO, the imaging of the black hole in M87 by the Event Horizon Telescope, and high- precision astrometry of the Galactic Center at close to the event horizon scale by the GRAVITY experiment motivate the development of numerical source models that solve the equations of general relativistic magnetohydrodynamics (GRMHD). Here we compare GRMHD solutions for the evolution of a magnetized accretion flow where turbulence is promoted by the magnetorotational instability from a set of nine GRMHD codes: Athena++, BHAC, Cosmos++, ECHO, H-AMR, iharm3D, HARM-Noble, IllinoisGRMHD, and KORAL. Agreement among the codes improves as resolution increases, as measured by a consistently applied, specially developed set of code performance metrics. We conclude that the community of GRMHD codes is mature, capable, and consistent on these test problems.

relativistic processes↗

The high energy Universe at ultra-high resolution: the power and promise of X-ray interferometry

We propose the development of X-ray interferometry (XRI), to reveal the Universe at high energies with ultra-high spatial resolution. With baselines which can be accommodated on a single spacecraft, XRI can reach 100 μ as resolution at 10 Å (1.2 keV) and 20 μ as at 2 Å (6 keV), enabling imaging and imaging-spectroscopy of (for example) X-ray coronae of nearby accreting supermassive black holes (SMBH) and the SMBH ‘shadow’; SMBH accretion flows and outflows; X-ray binary winds and orbits; stellar coronae within ∼100 pc and many exoplanets which transit across them. For sufficiently luminous sources XRI will resolve sub-pc scales across the entire observable Universe, revealing accreting binary SMBHs and enabling trigonometric measurements of the Hubble constant with X-ray light echoes from quasars or explosive transients. A multi-spacecraft ‘constellation’ interferometer would resolve well below 1 μ as, enabling SMBH event horizons to be resolved in many active galaxies and the detailed study of the effects of strong field gravity on the dynamics and emission from accreting gas close to the black hole.

Phil Uttley↗

An X-ray Interferometry Concept for the ESA Voyage 2050 Programme

We have proposed the development of X-ray interferometry as part of ESA’s Voyage 2050 programme, to reveal the universe at high energies with ultra-high spatial resolution. With only a 1 m baseline, which could be accommodated on a single spacecraft, X-ray interferometry can reach 100μas resolution at 10 ̊A (1.24 keV) and exceed that of the Event Horizon Telescope at 2 ̊A (6.2 keV). A multi-spacecraft ‘constellation’ interferometer would resolve well below 1μas. Here we focus on the single-spacecraft interferometer design and discuss the process of fringe detection and image reconstruction from multiple baselines, showing simulated images of testcases from our Voyage 2050 White Paper. We also discuss the challenges and feasibility of reaching the technical requirements needed for a single-spacecraft interferometer. Most key requirements are already feasible or within easy reach. Besides a ground-based testbed, covered elsewhere in these proceedings, the most important areas for development include large format, small-pixel X-ray detectors and pointing which is stable or can be reconstructed to tens of μas precision.

Phil Uttley↗