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Highest Resolutions (highest + resolution)
Selected AbstractsProtein separations using polyelectrolyte multilayer coatings with molecular micelles in open tubular capillary electrochromatographyELECTROPHORESIS, Issue 4 2008Candace A. Luces Abstract Novel polyelectrolyte multilayer (PEM) coatings for enhanced protein separations in open tubular CEC (OT-CEC) are reported. Use of four cationic polymers (poly- L -lysine, poly- L -ornithine, poly- L -lysine-serine, and poly- L -glutamic acid-lysine), and three anionic molecular micelles, sodium poly(N -undecanoyl- L -leucyl-alaninate) (poly- L -SULA), sodium poly(N -undecanoyl- L -leucyl-valinate) (poly- L -SULV), and sodium poly(undecylenic sulfate) (poly-SUS) were investigated in PEM coatings for protein separations. The simultaneous effects of cationic polymer concentration, number of bilayers, temperature, applied voltage, and pH of the BGE on the separation of four basic proteins (,-chymotrypsinogen A, lysozyme, ribonuclease A, and cytochrome c) were analyzed using a Box Behnken experimental design. The influence of NaCl on the run-to-run reproducibility was investigated for PEM coatings containing each cationic polymer. All coatings exhibited excellent reproducibilities with a %RSD of the EOF less than 1% in the presence of NaCl. Optimal conditions were dependent on both the cationic and anionic polymers used in the PEM coatings. Poly- L -glutamic acid-lysine produced the highest resolution and longest migration time. The use of molecular micelles to form PEM coatings resulted in better separations than single cationic coatings. Chiral poly- L -SULA and poly- L -SULV resulted in higher protein resolutions as compared to the achiral, poly-SUS. Furthermore, the use of poly- L -SULV reversed the elution order of lysozyme and cytochrome c when compared to poly- L -SULA and poly-SUS. [source] Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assayENVIRONMENTAL TOXICOLOGY, Issue 6 2008Alaa G. M. Osman Abstract Increasing lead contamination in Egyptian ecosystems and high lead concentrations in food items have raised concern for human health and stimulated studies on monitoring ecotoxicological impact of lead-caused genotoxicity. In this work, the alkaline comet assay was modified for monitoring DNA strand breakage in sensitive early life stages of the African catfish Clarias gariepinus. Following exposure to 100, 300, and 500 ,g/L lead nitrate, DNA strand breakage was quantified in embryos at 30, 48, 96, 144, and 168 h post-fertilization (PFS). For quantitative analysis, four commonly used parameters (tail % DNA, %TDNA; head % DNA, %HDNA; tail length, TL; tail moment, TM) were analyzed in 96 nuclei (in triplicates) at each sampling point. The parameter %TDNA revealed highest resolution and lowest variation. A strong correlation between lead concentration, time of exposure, and DNA strand breakage was observed. Here, genotoxicity detected by comet assay preceded the manifested malformations assessed with conventional histology. Qualitative evaluation was carried out using five categories are as follows: undamaged (%TDNA , 10%), low damaged (10% < %TDNA , 25%), median damaged (25 < %TDNA , 50%), highly damaged (50 < %TDNA , 75%), and extremely damaged (%TDNA > 75%) nuclei confirming a dose and time-dependent shift towards increased frequencies of highly and extremely damaged nuclei. A protective capacity provided by a hardened chorion is a an interesting finding in this study as DNA damage in the prehatching stages 30 h-PFS and 48 h-PFS was low in all treatments (qualitative and quantitative analyses). These results clearly show that the comet assay is a sensitive tool for the detection of genotoxicity in vulnerable early life stages of the African catfish and is a method more sensitive than histological parameters for monitoring genotoxic effects. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2008. [source] Spatially Resolved Very Large Array 74 MHz Observations Toward the Galactic CenterASTRONOMISCHE NACHRICHTEN, Issue S1 2003C. L. Brogan Abstract We present the highest resolution and sensitivity low frequency image (<300 MHz) of the Galactic center to date using the Very Large Array at 74 MHz in its A, B, C, & D configurations. The resulting images have a resolution of 2.1, × 1.2, and a dynamic range of ,400 From this data we have been able to identify a region of enhanced 74 MHz emission about 5° in extent that is coincident with the high density molecular gas surrounding the Galactic center known as the Central Molecular Zone. In addition to giving an unprecedented view of the extended nonthermal emission surrounding the Galactic center, the 74 MHz image shows deep free-free absorption across the Galactic center itself, as well as, part of the Galactic center radio lobe, and a number of H II regions in the field. This absorption is due to ionized thermal gas in front of, or in some cases embedded in, the nonthermal Galactic center (GC) emission. Such absorption allows us to unambiguously place some of the H II regions in the direction of the GC along the line of sight for the first time. The morphology, nature, and relationship to the Galactic center of the 74 MHz absorption and emission is discussed. [source] The structure of PhaZ7 at atomic (1.2,Å) resolution reveals details of the active site and suggests a substrate-binding modeACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 6 2010Sachin Wakadkar Poly-(R)-hydroxyalkanoates (PHAs) are bacterial polyesters that are degraded by a group of enzymes known as PHA depolymerases. Paucimonas lemoignei PhaZ7 depolymerase is the only extracellular depolymerase that has been described as being active towards amorphous PHAs. A previously determined crystal structure of PhaZ7 revealed an ,/,-hydrolase fold and a Ser-His-Asp catalytic triad. In order to address questions regarding the catalytic mechanism and substrate binding, the atomic resolution structure of PhaZ7 was determined after cocrystallization with the protease inhibitor PMSF. The reported structure has the highest resolution (1.2,Å) of currently known depolymerase structures and shows a sulfur dioxide molecule covalently attached to the active-site residue Ser136. Structural comparison with the free PhaZ7 structure (1.45,Å resolution) revealed no major changes in the active site, suggesting a preformed catalytic triad. The oxyanion hole was found to be formed by the amide groups of Met137 and Asn49. Nine well ordered water molecules were located in the active site. Manual docking of a substrate trimer showed that the positions of these water molecules coincide well with the substrate atoms. It is proposed that these water molecules are displaced upon binding of the substrate. Furthermore, conformational changes were identified after comparison with a previously determined PhaZ7 dimer structure in a different space group. The changes were located in surface loops involved in dimer formation, indicating some flexibility of these loops and their possible involvement in polyester binding. [source] Structure of Helicobacter pyloril -asparaginase at 1.4,Å resolutionACTA CRYSTALLOGRAPHICA SECTION D, Issue 12 2009Prathusha Dhavala Bacterial l -asparaginases have been used in the treatment of childhood acute lymphoblastic leukaemia for over 30,years. Their therapeutic effect is based on their ability to catalyze the conversion of l -asparagine, an essential amino acid in certain tumours, to l -aspartic acid and ammonia. Two l -asparaginases, one from Escherichia coli and the other from Erwinia chrysanthemi, have been widely employed in clinical practice as anti-leukaemia drugs. However, l -asparaginases are also able to cause severe side effects owing to their intrinsic glutaminase activity. Helicobacter pyloril -asparaginase (HpA) has been reported to have negligible glutaminase activity. To gain insight into the properties of HpA, its crystal structure in the presence of l -aspartate was determined to 1.4,Å resolution, which is one of the highest resolutions obtained for an l -asparaginase structure. The final structure has an Rcryst of 12.6% (Rfree = 16.9%) with good stereochemistry. A detailed analysis of the active site showed major differences in the active-site flexible loop and in the 286,297 loop from the second subunit, which is involved in active-site formation. Accordingly, Glu289, Asn255 and Gln63 are suggested to play roles in modulating the accessibility of the active site. Overall, the structural comparison revealed that HpA has greater structural similarity to E. colil -asparaginase than to any other l -asparaginase, including Er. carotovoral -asparaginase, despite the fact that the latter is also characterized by low glutaminase activity. [source] High-fidelity spectroscopy at the highest resolutionsASTRONOMISCHE NACHRICHTEN, Issue 5 2010D. Dravins Abstract High-fidelity spectroscopy presents challenges for both observations and in designing instruments. High-resolution and high-accuracy spectra are required for verifying hydrodynamic stellar atmospheres and for resolving intergalactic absorption-line structures in quasars. Even with great photon fluxes from large telescopes with matching spectrometers, precise measurements of line profiles and wavelength positions encounter various physical, observational, and instrumental limits. The analysis may be limited by astrophysical and telluric blends, lack of suitable lines, imprecise laboratory wavelengths, or instrumental imperfections. To some extent, such limits can be pushed by forming averages over many similar spectral lines, thus averaging away small random blends and wavelength errors. In situations where theoretical predictions of lineshapes and shifts can be accurately made (e.g., hydrodynamic models of solar-type stars), the consistency between noisy observations and theoretical predictions may be verified; however this is not feasible for, e.g., the complex of intergalactic metal lines in spectra of distant quasars, where the primary data must come from observations. To more fully resolve lineshapes and interpret wavelength shifts in stars and quasars alike, spectral resolutions on order R = 300 000 or more are required; a level that is becoming (but is not yet) available. A grand challenge remains to design efficient spectrometers with resolutions approaching R = 1 000 000 for the forthcoming generation of extremely large telescopes (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] A high-resolution modelling case study of a severe weather event over New ZealandATMOSPHERIC SCIENCE LETTERS, Issue 3 2008Stuart Webster Abstract In this article, the ability of the Met Office Unified Model to simulate the severe weather over the South Island of New Zealand, on 8 January 2004 is investigated. Simulations were run at horizontal resolutions of between 60 and 1 km. The modelled broad-scale rainfall and wind features, most notably the area-averaged accumulated rainfall, were found to converge with resolution. At the highest resolutions, all the observed rainfall and wind features of this event were captured well by the model. Even the 12-km-resolution model is able to resolve the broad elongated ridge-like structure of the Southern Alps and qualitatively capture the main features of the rainfall and wind fields. Copyright © 2008 Royal Meteorological Society and Crown Copyright 2008, published by John Wiley & Sons, Ltd. [source] High-quality crystals of human haematopoietic prostaglandin D synthase with novel inhibitorsACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2010Sachiko Takahashi Human haematopoietic prostaglandin D synthase (H-PGDS; EC 5.3.99.2) produces prostaglandin D2, an allergic and inflammatory mediator, in mast cells and Th2 cells. H-PGDS has been crystallized with novel inhibitors with half-maximal inhibitory concentrations (IC50) in the low nanomolar range by the counter-diffusion method onboard the Russian Service Module on the International Space Station. The X-ray diffraction of a microgravity-grown crystal of H-PGDS complexed with an inhibitor with an IC50 value of 50,nM extended to 1.1,Å resolution at 100,K using SPring-8 synchrotron radiation, which is one of the highest resolutions obtained to date for this protein. [source] Cloning, expression, purification, crystallization and preliminary structure determination of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphateACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 9 2006D. Aragão The cloning, expression, purification, crystallization and preliminary crystallographic analysis of glucose-1-phosphate uridylyltransferase (UgpG) from Sphingomonas elodea ATCC 31461 bound to glucose-1-phosphate are reported. Diffraction data sets were obtained from seven crystal forms in five different space groups, with highest resolutions ranging from 4.20 to 2.65,Å. The phase problem was solved for a P21 crystal form using multiple isomorphous replacement with anomalous scattering from an osmium derivative and a SeMet derivative. The best native crystal in space group P21 has unit-cell parameters a = 105.5, b = 85.7, c = 151.8,Å, , = 105.2°. Model building and refinement are currently under way. 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