Expression of active human sialyltransferase ST6GalNAcI in Escherichia coli

Georgios Skretas, Sean Carroll, and George Georgiou are with the Department of Chemical Engineering, University of Texas at Austin, Austin, TX 78712, USA -- George Georgiou is with the Department of Biomedical Engineering, University of Texas at Austin and the Section of Microbiology and Molecular Genetics, University of Texas at Austin, Austin, TX 78712, USA -- Georgios Skretas and George Georgiou are with the Institute for Cellular and Molecular Biology, University of Texas at Austin, Austin, TX 78712, USA -- Shawn DeFrees, Karl F. Johnson, and Marc F. Schwartz are with Neose Technologies Inc, 102 Rock Road, Horsham, PA, 19044, USABackground: The presence of terminal, surface-exposed sialic acid moieties can greatly enhance the in vivo half-life of glycosylated biopharmaceuticals and improve their therapeutic efficacy. Complete and homogeneous sialylation of glycoproteins can be efficiently performed enzymically in vitro but this process requires large amounts of catalytically active sialyltransferases. Furthermore, standard microbial hosts used for large-scale production of recombinant enzymes can only produce small quantities of glycosyltransferases of animal origin, which lack catalytic activity. Results and conclusion: In this work, we have expressed the human sialyltransferase ST6GalNAc I (ST6), an enzyme that sialylates O-linked glycoproteins, in Escherichia coli cells. We observed that wild-type bacterial cells are able to produce only very small amounts of soluble ST6 enzyme. We have found, however, that engineered bacterial strains which possess certain types of oxidative cytoplasm or which co-express the molecular chaperones/co-chaperones trigger factor, DnaK/DnaJ, GroEL/GroES, and Skp, can produce greatly enhanced amounts of soluble ST6. Furthermore, we have developed a novel high-throughput assay for the detection of sialyltransferase activity and used it to demonstrate that the bacterially expressed ST6 enzyme is active and able to transfer sialic acid onto a desialylated O-glycoprotein, bovine submaxillary mucin. To the best of our knowledge, this is the first example of expression of active human sialyltransferase in bacteria. This system may be used as a starting point for the evolution of sialyltransferases with better expression characteristics or altered donor/acceptor specificities.Chemical EngineeringBiomedical EngineeringInstitute for Cellular and Molecular [email protected]

Estimating the sensitivity of wide-parameter-space searches for gravitational-wave pulsars

This paper presents an in-depth study of how to estimate the sensitivity of searches for gravitational-wave pulsars -- rapidly-rotating neutron stars which emit quasi-sinusoidal gravitational waves. It is particularly concerned with searches over a wide range of possible source parameters, such as searches over the entire sky and broad frequency bands. Traditional approaches to estimating the sensitivity of such searches use either computationally-expensive Monte Carlo simulations, or analytic methods which sacrifice accuracy by making an unphysical assumption about the population of sources being searched for. This paper develops a new, analytic method of estimating search sensitivity which does not rely upon this unphysical assumption. Unlike previous analytic methods, the new method accurately predicts the sensitivity obtained using Monte Carlo simulations, while avoiding their computational expense. The change in estimated sensitivity due to properties of the search template bank, and the geographic configuration of the gravitational wave detector network, are also investigated.Comment: 16 figures, 2 tables, REVTeX 4.1; minor typos corrected from v2, updated reference

The Small Magellanic Cloud Investigation of Dust and Gas Evolution (SMIDGE): The Dust Extinction Curve from Red Clump Stars

We use Hubble Space Telescope (HST) observations of red clump stars taken as part of the Small Magellanic Cloud Investigation of Dust and Gas Evolution (SMIDGE) program to measure the average dust extinction curve in a ~ 200 pc x 100 pc region in the southwest bar of the Small Magellanic Cloud (SMC). The rich information provided by our 8-band ultra-violet through near-infrared photometry allows us to model the color-magnitude diagram of the red clump accounting for the extinction curve shape, a log-normal distribution of $A_{V}$, and the depth of the stellar distribution along the line of sight. We measure an extinction curve with $R_{475} = A_{475}/(A_{475}-A_{814})$ = 2.65 $\pm$ 0.11. This measurement is significantly larger than the equivalent values of published Milky Way $R_{V}$ = 3.1 ($R_{475} = 1.83$) and SMC Bar $R_{V}$ = 2.74 ($R_{475} = 1.86$) extinction curves. Similar extinction curve offsets in the Large Magellanic Cloud (LMC) have been interpreted as the effect of large dust grains. We demonstrate that the line-of-sight depth of the SMC (and LMC) introduces an apparent "gray" contribution to the extinction curve inferred from the morphology of the red clump. We show that no gray dust component is needed to explain extinction curve measurements when a full-width half-max depth of 10 $\pm$ 2 kpc in the stellar distribution of the SMC (5 $\pm$ 1 kpc for the LMC) is considered, which agrees with recent studies of Magellanic Cloud stellar structure. The results of our work demonstrate the power of broad-band HST imaging for simultaneously constraining dust and galactic structure outside the Milky Way.Comment: 16 pages, 12 figures, 5 tables. Accepted for publication in Ap

Toward Agent-based Modeling of the U.S. Department of Defense Acquisition System

The systems development, procurement and sustainment of a nation\u27s military equipment is vital to its national interests, but the process is complex, constantly changing and highly adaptive, as well as time consuming and costly. The U.S. Department of Defense (DoD) expends both large amounts of capital and manpower to equip its armed forces. This research seeks to identify opportunities to gain better insight into the functioning of the defense acquisition system, building on previous simulations. A case is made that the DoD Requirements, Planning Acquisition, Technology and Logistics System is a complex adaptive system that has characteristics appropriate for exploration with agent-based modeling. This paper reviews relevant literature, existing models and presents preliminary analysis on an agent-based simulation of the DoD acquisition system. This research finds that agent-based modeling can provide insights to inform new acquisition theory

Wavelength Tunability of Ion-bombardment Induced Ripples on Sapphire

A study of ripple formation on sapphire surfaces by 300-2000 eV Ar+ ion bombardment is presented. Surface characterization by in-situ synchrotron grazing incidence small angle x-ray scattering and ex-situ atomic force microscopy is performed in order to study the wavelength of ripples formed on sapphire (0001) surfaces. We find that the wavelength can be varied over a remarkably wide range-nearly two orders of magnitude-by changing the ion incidence angle. Within the linear theory regime, the ion induced viscous flow smoothing mechanism explains the general trends of the ripple wavelength at low temperature and incidence angles larger than 30. In this model, relaxation is confined to a few-nm thick damaged surface layer. The behavior at high temperature suggests relaxation by surface diffusion. However, strong smoothing is inferred from the observed ripple wavelength near normal incidence, which is not consistent with either surface diffusion or viscous flow relaxation.Comment: Revtex4, 19 pages, 10 figures with JPEG forma

Extinction Corrected Star Formation Rates Empirically Derived from Ultraviolet-Optical Colors

Using a sample of galaxies from the Sloan Digital Sky Survey spectroscopic catalog with measured star-formation rates (SFRs) and ultraviolet (UV) photometry from the GALEX Medium Imaging Survey, we derived empirical linear correlations between the SFR to UV luminosity ratio and the UV-optical colors of blue sequence galaxies. The relations provide a simple prescription to correct UV data for dust attenuation that best reconciles the SFRs derived from UV and emission line data. The method breaks down for the red sequence population as well as for very blue galaxies such as the local ``supercompact'' UV luminous galaxies and the majority of high redshift Lyman Break Galaxies which form a low attenuation sequence of their own.Comment: 20 pages, 11 figures, accepted for publication in the ApJS GALEX special issu

The Star Formation and Extinction Co-Evolution of UV-Selected Galaxies over 0.05<z<1.2

We use a new stacking technique to obtain mean mid IR and far IR to far UV flux ratios over the rest near-UV/near-IR color-magnitude diagram. We employ COMBO-17 redshifts and COMBO-17 optical, GALEX far and near UV, Spitzer IRAC and MIPS Mid IR photometry. This technique permits us to probe infrared excess (IRX), the ratio of far IR to far UV luminosity, and specific star formation rate (SSFR) and their co-evolution over two orders of magnitude of stellar mass and redshift 0.1<z<1.2. We find that the SSFR and the characteristic mass (M_0) above which the SSFR drops increase with redshift (downsizing). At any given epoch, IRX is an increasing function of mass up to M_0. Above this mass IRX falls, suggesting gas exhaustion. In a given mass bin below M_0 IRX increases with time in a fashion consistent with enrichment. We interpret these trends using a simple model with a Schmidt-Kennicutt law and extinction that tracks gas density and enrichment. We find that the average IRX and SSFR follows a galaxy age parameter which is determined mainly by the galaxy mass and time since formation. We conclude that blue sequence galaxies have properties which show simple, systematic trends with mass and time such as the steady build-up of heavy elements in the interstellar media of evolving galaxies and the exhaustion of gas in galaxies that are evolving off the blue sequence. The IRX represents a tool for selecting galaxies at various stages of evolution.Comment: Accepted for publication in GALEX Special Ap.J.Suppl., December, 200

Ultraviolet through Infrared Spectral Energy Distributions from 1000 SDSS Galaxies: Dust Attenuation

The meaningful comparison of models of galaxy evolution to observations is critically dependent on the accurate treatment of dust attenuation. To investigate dust absorption and emission in galaxies we have assembled a sample of ~1000 galaxies with ultraviolet (UV) through infrared (IR) photometry from GALEX, SDSS, and Spitzer and optical spectroscopy from SDSS. The ratio of IR to UV emission (IRX) is used to constrain the dust attenuation in galaxies. We use the 4000A break as a robust and useful, although coarse, indicator of star formation history (SFH). We examine the relationship between IRX and the UV spectral slope (a common attenuation indicator at high-redshift) and find little dependence of the scatter on 4000A break strength. We construct average UV through far-IR spectral energy distributions (SEDs) for different ranges of IRX, 4000A break strength, and stellar mass (M_*) to show the variation of the entire SED with these parameters. When binned simultaneously by IRX, 4000A break strength, and M_* these SEDs allow us to determine a low resolution average attenuation curve for different ranges of M_*. The attenuation curves thus derived are consistent with a lambda^{-0.7} attenuation law, and we find no significant variations with M_*. Finally, we show the relationship between IRX and the global stellar mass surface density and gas-phase-metallicity. Among star forming galaxies we find a strong correlation between IRX and stellar mass surface density, even at constant metallicity, a result that is closely linked to the well-known correlation between IRX and star-formation rate.Comment: 12 pages, 8 figures, 2 tables, appearing in the Dec 2007 GALEX special issue of ApJ Supp (29 papers