Crystallographic Study Of The Phosphoethanolamine Transferase EptC required For Polymyxin Resistance And Motility In Campylobacter jejuni

The foodborne enteric pathogen Campylobacter jejuni decorates a variety of its cell-surface structures with phosphoethanolamine (pEtN). Modifying lipid A with pEtN promotes cationic antimicrobial peptide resistance, whereas post-translationally modifying the flagellar rod protein FlgG with pEtN promotes flagellar assembly and motility, which are processes that are important for intestinal colonization. EptC, the pEtN transferase required for all known pEtN cell-surface modifications in C. jejuni, is a predicted inner-membrane metalloenzyme with a five-helix N-terminal transmembrane domain followed by a soluble sulfatase-like catalytic domain in the periplasm. The atomic structure of the catalytic domain of EptC (cEptC) was crystallized and solved to a resolution of 2.40 angstrom. cEptC adopts the alpha/beta/alpha fold of the sulfatase protein family and harbors a zinc-binding site. A phosphorylated Thr266 residue was observed that was hypothesized to mimic a covalent pEtN-enzyme intermediate. The requirement for Thr266 as well as the nearby residues Asn308, Ser309, His358 and His440 was ascertained via in vivo activity assays on mutant strains. The results establish a basis for the design of pEtN transferase inhibitors.National Institutes of Health (grants AI064184, AI076322, GM106112Army Research Office (grantW911NF-12-1-0390)College of Natural SciencesOffice of the Executive Vice President and ProvostInstitute for Cellular and Molecular Biology at the University of Texas at AustinUS DOE DE-AC02-06CH11357National Institute of General Medical SciencesHoward Hughes Medical InstituteOffice of Science, Office of Basic Energy Sciences of the US Department of Energy DE-AC02-05CH11231Maria Person and the Proteomics Facility at the University of Texas at Austin ES007784 (CRED) and RP110782 (CPRIT)Molecular Bioscience

Molecular weight of hydroxyethyl starch: is there an effect on blood coagulation and pharmacokinetics?

Background. The development of hydroxyethyl starches (HES) with low impact on blood coagulation but higher volume effect compared with the currently used HES solutions is of clinical interest. We hypothesized that high molecular weight, low-substituted HES might possess these properties. Methods. Thirty pigs were infused with three different HES solutions (20 ml kg−1) with the same degree of molar substitution (0.42) but different molecular weights (130, 500 and 900 kDa). Serial blood samples were taken over 24 h and blood coagulation was assessed by Thromboelastograph® analysis and analysis of plasma coagulation. In addition, plasma concentration and in vivo molecular weight were determined and pharmacokinetic data were computed based on a two-compartment model. Results. Thromboelastograph analysis and plasma coagulation tests did not reveal a more pronounced alteration of blood coagulation with HES 500 and HES 900 compared with HES 130. In contrast, HES 500 and HES 900 had a greater area under the plasma concentration-time curve [1542 (142) g min litre−1, P<0.001, 1701 (321) g min litre−1, P<0.001] than HES 130 [1156 (223) g min litre−1] and alpha half life (\batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} $\mathrm{t}_{{\alpha}}^{{\frac{1}{2}}}$ \end{document}) was longer for HES 500 [53.8 (8.6) min, P<0.01] and HES 900 [57.1 (12.3) min, P<0.01] than for HES 130 [39.9 (10.7) min]. Beta half life (\batchmode \documentclass[fleqn,10pt,legalpaper]{article} \usepackage{amssymb} \usepackage{amsfonts} \usepackage{amsmath} \pagestyle{empty} \begin{document} $\mathrm{t}_{{\beta}}^{{\frac{1}{2}}}$ \end{document}), however, was similar for all three types of HES [from 332 (100) to 381 (63) min]. Conclusions. In low-substituted HES, molecular weight is not a key factor in compromising blood coagulation. The longer initial intravascular persistence of high molecular weight low-substituted HES might result in a longer lasting volume effec

Blind Normalization of Speech From Different Channels

We show how to construct a channel-independent representation of speech that has propagated through a noisy reverberant channel. This is done by blindly rescaling the cepstral time series by a non-linear function, with the form of this scale function being determined by previously encountered cepstra from that channel. The rescaled form of the time series is an invariant property of it in the following sense: it is unaffected if the time series is transformed by any time-independent invertible distortion. Because a linear channel with stationary noise and impulse response transforms cepstra in this way, the new technique can be used to remove the channel dependence of a cepstral time series. In experiments, the method achieved greater channel-independence than cepstral mean normalization, and it was comparable to the combination of cepstral mean normalization and spectral subtraction, despite the fact that no measurements of channel noise or reverberations were required (unlike spectral subtraction).Comment: 25 pages, 7 figure

Noise Reduction Using Wavelet Thresholding of Multitaper Estimators and Geometric Approach to Spectral Subtraction for Speech Coding Strategy

ObjectivesNoise reduction using wavelet thresholding of multitaper estimators (WTME) and geometric approach to spectral subtraction (GASS) can improve speech quality of noisy sound for speech coding strategy. This study used Perceptual Evaluation of Speech Quality (PESQ) to assess the performance of the WTME and GASS for speech coding strategy.MethodsThis study included 25 Mandarin sentences as test materials. Environmental noises including the air-conditioner, cafeteria and multi-talker were artificially added to test materials at signal to noise ratio (SNR) of -5, 0, 5, and 10 dB. HiRes 120 vocoder WTME and GASS noise reduction process were used in this study to generate sound outputs. The sound outputs were measured by the PESQ to evaluate sound quality.ResultsTwo figures and three tables were used to assess the speech quality of the sound output of the WTME and GASS.ConclusionThere is no significant difference between the overall performance of sound quality in both methods, but the geometric approach to spectral subtraction method is slightly better than the wavelet thresholding of multitaper estimators

High temperature strength retention of Cu/Nb nanolaminates through dynamic strain ageing

The mechanical properties of Cu/Nb metallic nanolaminates with different layer thickness (7, 16, 34 and 63 nm) were studied by means of micropillar compression tests from room temperature to 400 C. Both strain-rate jump and constant strain rate tests were carried out and they showed evidence of dynamic strain ageing in the nanolaminates with 7, 16 and 34 nm layer thickness deformed at 200 C. Dynamic strain ageing was accompanied by a reduction of the strain rate sensitivity to 0, high strength retention at 200 C and the development of shear localization of the deformation at low strains (5%-6%) that took place along the Nb layers in the nanolaminates. Atom probe tomography of the deformed specimens revealed the presence of O in solid solution in the Nb layers but not in the Cu layers. Thus, diffusion of O atoms to the mobile dislocations in Nb was found to be the origin of the dynamic strain ageing in the Cu/Nb nanolaminates around 200 C. This mechanism was not found at higher temperatures (400 C) because deformation was mainly controlled by stress-assisted diffusion in the Cu layers. This discovery shows a novel strategy to enhance the strength retention at high temperature of metallic nanolaminates through dynamic strain ageing of one the phases

Time-Resolved Measurement of Interatomic Coulombic Decay in Ne_2

The lifetime of interatomic Coulombic decay (ICD) [L. S. Cederbaum et al., Phys. Rev. Lett. 79, 4778 (1997)] in Ne_2 is determined via an extreme ultraviolet pump-probe experiment at the Free-Electron Laser in Hamburg. The pump pulse creates a 2s inner-shell vacancy in one of the two Ne atoms, whereupon the ionized dimer undergoes ICD resulting in a repulsive Ne^{+}(2p^{-1}) - Ne^{+}(2p^{-1}) state, which is probed with a second pulse, removing a further electron. The yield of coincident Ne^{+} - Ne^{2+} pairs is recorded as a function of the pump-probe delay, allowing us to deduce the ICD lifetime of the Ne_{2}^{+}(2s^{-1}) state to be (150 +/- 50) fs in agreement with quantum calculations.Comment: 5 pages, 3 figures, accepted by PRL on July 11th, 201

Multimedia Vocabularies on the Semantic Web

This document gives an overview on the state-of-the-art of multimedia metadata formats. Initially, practical relevant vocabularies for developers of Semantic Web applications are listed according to their modality scope. In the second part of this document, the focus is set on the integration of the multimedia vocabularies into the Semantic Web, that is to say, formal representations of the vocabularies are discussed