The interaction of oxygen with small gold clusters

Presented in this work are the results of a quantum chemical study of oxygen adsorption on small Aun and Au−n (n=2,3) clusters. Density functional theory(DFT), second order perturbation theory (MP2), and singles and doubles coupled clustertheory with perturbative triples [CCSD(T)] methods have been used to determine the geometry and the binding energy of oxygen to Aun. The multireference character of the wave functions has been studied using the complete active space self-consistent field method. There is considerable disagreement between the oxygen binding energies provided by CCSD(T) calculations and those obtained with DFT. The disagreement is often qualitative, with DFT predicting strong bonds where CCSD(T) predicts no bonds or structures that are bonded but have energies that exceed those of the separated components. The CCSD(T) results are consistent with experimental measurements, while DFT calculations show, at best, a qualitative agreement. Finally, the lack of a regular pattern in the size and the sign of the errors [as compared to CCSD(T)] is a disappointing feature of the DFT results for the present system: it is not possible to give a simple rule for correcting the DFT predictions (e.g., a useful rule would be that DFT predicts stronger binding of O2 by about 0.3 eV). It is likely that the errors in DFT appear not because of gold, but because oxygen binding to a metal cluster is a particularly difficult problem.This work was supported by AFOSR through a DURINT grant

Changes in Health and Physical Fitness Parameters After 6 Months of High-intensity Group Exercise in Firefighters: Preliminary Data

The physical demands of firefighting require the men and women employed in this profession to be in optimal physical condition to perform their jobs proficiently, as well as to mitigate the risk of injury. Every year, the city of Addison, TX, budgets many thousands of dollars to the compensation plan for first responders. Most of the funding is used for rehabilitation services due to work related injuries. PURPOSE: While many of these injuries are unavoidable due to the inherent risks of the profession, ensuring proper physical fitness is one of the most effective methods to reduce many of these impairments. The purpose of this preliminary study was to characterize health and fitness parameters in 18 professional firefighters from the city of Addison, TX, prior to a 6-month training program. METHODS: Upon arrival, all participants underwent testing in the following order: body composition, range of motion, anaerobic power, muscular endurance, and cardiorespiratory fitness. All participants also completed a detailed health history questionnaire and answered questions specifically addressing chronic low back pain. RESULTS: The following values were attained from testing: total body fat: 30.1±9.7%; flexibility: 24.9±6.3 cm; peak power: 1068.7±272.9 W; mean power: 636.9±143.4 W; time to peak power 0.57±0.35 sec; pushup: 28.8±14.9; curl up: 22.1±15.8; VO2max: 34.1±5.1 ml/kg/min. CONCLUSION: Based off the comparison of reported means and ACSM’s fitness norms, it can be concluded that improvements are necessary in body composition, muscular endurance, range of motion and cardiorespiratory fitness. With improvements in these physiological variables, tactical performance may be optimally performed in a safer manner

A Spectroscopic Survey of Faint Quasars in the SDSS Deep Stripe: I. Preliminary Results from the Co-added Catalog

In this paper we present the first results of a deep spectroscopic survey of faint quasars in the Sloan Digital Sky Survey (SDSS) Southern Survey, a deep survey carried out by repeatedly imaging a 270 deg^2 area. Quasar candidates were selected from the deep data with good completeness over 0<z<5, and 2 to 3 magnitudes fainter than the SDSS main survey. Spectroscopic follow-up was carried out on the 6.5m MMT with Hectospec. The preliminary sample of this SDSS faint quasar survey (hereafter SFQS) covers ~ 3.9 deg^2, contains 414 quasars, and reaches g=22.5. The overall selection efficiency is ~ 66% (~ 80% at g<21.5); the efficiency in the most difficult redshift range (2<z<3) is better than 40%. We use the 1/V_{a} method to derive a binned estimate of the quasar luminosity function (QLF) and model the QLF using maximum likelihood analysis. The best model fits confirm previous results showing that the QLF has steep slopes at the bright end and much flatter slopes (-1.25 at z<2.0 and -1.55 at z>2.0) at the faint end, indicating a break in the QLF slope. Using a luminosity-dependent density evolution model, we find that the quasar density at M_{g}<-22.5 peaks at z~2, which is later in cosmic time than the peak of z~2.5 found from surveys of more luminous objects. The SFQS QLF is consistent with the results of the 2dF QSO Redshift Survey, the SDSS, and the 2dF-SDSS LRG and QSO Survey, but probes fainter quasars. We plan to obtain more quasars from future observations and establish a complete faint quasar sample with more than 1000 objects over 10 deg^2.Comment: 25 pages, 13 figures, accepted for publication in A

Advances in Understanding Carboxysome Assembly in \u3ci\u3eProchlorococcus\u3c/i\u3e and \u3ci\u3eSynechococcus\u3c/i\u3e Implicate CsoS2 as a Critical Component

The marine Synechococcus and Prochlorococcus are the numerically dominant cyanobacteria in the ocean and important in global carbon fixation. They have evolved a CO2-concentrating-mechanism, of which the central component is the carboxysome, a self-assembling proteinaceous organelle. Two types of carboxysome, α and β, encapsulating form IA and form IB d-ribulose-1,5-bisphosphate carboxylase/oxygenase, respectively, differ in gene organization and associated proteins. In contrast to the β-carboxysome, the assembly process of the α-carboxysome is enigmatic. Moreover, an absolutely conserved α-carboxysome protein, CsoS2, is of unknown function and has proven recalcitrant to crystallization. Here, we present studies on the CsoS2 protein in three model organisms and show that CsoS2 is vital for α-carboxysome biogenesis. The primary structure of CsoS2 appears tripartite, composed of an N-terminal, middle (M)-, and C-terminal region. Repetitive motifs can be identified in the N- and M-regions. Multiple lines of evidence suggest CsoS2 is highly flexible, possibly an intrinsically disordered protein. Based on our results from bioinformatic, biophysical, genetic and biochemical approaches, including peptide array scanning for protein-protein interactions, we propose a model for CsoS2 function and its spatial location in the α-carboxysome. Analogies between the pathway for β-carboxysome biogenesis and our model for α-carboxysome assembly are discussed

The cellular localization of human cytomegalovirus glycoprotein expression greatly influences the frequency and functional phenotype of specific CD4+ T cell responses

CMV infection is a significant cause of morbidity and mortality in immunocompromised individuals, and the development of a vaccine is of high priority. Glycoprotein B (gB) is a leading vaccine candidate but the glycoprotein H (gH) pentameric complex is now recognized as the major target for neutralizing Abs. However, little is known about the T cell immune response against gH and glycoprotein L (gL) and this is likely to be an important attribute for vaccine immunogenicity. In this study, we examine and contrast the magnitude and phenotype of the T cell immune response against gB, gH, and gL within healthy donors. gB-specific CD4(+) T cells were found in 95% of donors, and 29 epitopes were defined with gB-specific response sizes ranging from 0.02 to 2.88% of the CD4(+) T cell pool. In contrast, only 20% of donors exhibited a T cell response against gH or gL. Additionally, gB-specific CD4(+) T cells exhibited a more cytotoxic phenotype, with high levels of granzyme B expression. Glycoproteins were effectively presented following delivery to APCs but only gB-derived epitopes were presented following endogenous synthesis. gB expression was observed exclusively within vesicular structures colocalizing with HLA-DM whereas gH was distributed evenly throughout the cytoplasm. Grafting of the C-terminal domain from gB onto gH could not transfer this pattern of presentation. These results reveal that gB is a uniquely immunogenic CMV glycoprotein and this is likely to reflect its unique pattern of endogenous Ag presentation. Consideration may be required toward mechanisms that boost cellular immunity to gH and gL within future subunit vaccines

Ultra-Fast Bioorthogonal Spin-Labeling and Distance Measurements in Mammalian Cells Using Small, Genetically Encoded Tetrazine Amino Acids

Studying protein structures and dynamics directly in the cellular environments in which they function is essential to fully understand the molecular mechanisms underlying cellular processes. Site-directed spin-labeling (SDSL)—in combination with double electron–electron resonance (DEER) spectroscopy—has emerged as a powerful technique for determining both the structural states and the conformational equilibria of biomacromolecules. In-cell DEER spectroscopy on proteins in mammalian cells has thus far not been possible due to the notable challenges of spin-labeling in live cells. In-cell SDSL requires exquisite biorthogonality, high labeling reaction rates and low background signal from unreacted residual spin label. While the bioorthogonal reaction must be highly specific and proceed under physiological conditions, many spin labels display time-dependent instability in the reducing cellular environment. Additionally, high concentrations of spin label can be toxic. Thus, an exceptionally fast bioorthogonal reaction is required that can allow for complete labeling with low concentrations of spin-label prior to loss of signal. Here we utilized genetic code expansion to site-specifically encode a novel family of small, tetrazine-bearing non-canonical amino acids (Tet-v4.0) at multiple sites in green fluorescent protein (GFP) and maltose binding protein (MBP) expressed both in E. coli and in human HEK293T cells. We achieved specific and quantitative spin-labeling of Tet-v4.0-containing proteins by developing a series of strained trans-cyclooctene (sTCO)-functionalized nitroxides—including a gem-diethyl-substituted nitroxide with enhanced stability in cells—with rate constants that can exceed 106 M−1 s−1. The remarkable speed of the Tet-v4.0/sTCO reaction allowed efficient spin-labeling of proteins in live HEK293T cells within minutes, requiring only sub-micromolar concentrations of sTCO–nitroxide added directly to the culture medium. DEER recorded from intact cells revealed distance distributions in good agreement with those measured from proteins purified and labeled in vitro. Furthermore, DEER was able to resolve the maltose-dependent conformational change of Tet-v4.0-incorporated and spin-labeled MBP in vitro and successfully discerned the conformational state of MBP within HEK293T cells. We anticipate the exceptional reaction rates of this system, combined with the relatively short and rigid side chains of the resulting spin labels, will enable structure/function studies of proteins directly in cells, without any requirements for protein purification

A novel targeted/untargeted GC-Orbitrap metabolomics methodology applied to Candida albicans and Staphylococcus aureus biofilms

Introduction: Combined infections from Candida albicans and Staphylococcus aureus are a leading cause of death in the developed world. Evidence suggests that Candida enhances the virulence of Staphylococcus—hyphae penetrate through tissue barriers, while S. aureus tightly associates with the hyphae to obtain entry to the host organism. Indeed, in a biofilm state, C. albicans enhances the antimicrobial resistance characteristics of S. aureus. The association of these microorganisms is also associated with significantly increased morbidity and mortality. Due to this tight association we hypothesised that metabolic effects were also in evidence. Objectives: To explore the interaction, we used a novel GC-Orbitrap-based mass spectrometer, the Q Exactive GC, which combines the high peak capacity and chromatographic resolution of gas chromatography with the sub-ppm mass accuracy of an Orbitrap system. This allows the capability to leverage the widely available electron ionisation libraries for untargeted applications, along with expanding accurate mass libraries and targeted matches based around authentic standards. Methods: Optimised C. albicans and S. aureus mono- and co-cultured biofilms were analysed using the new instrument in addition to the fresh and spent bacterial growth media. Results: The targeted analysis experiment was based around 36 sugars and sugar phosphates, 22 amino acids and five organic acids. Untargeted analysis resulted in the detection of 465 features from fresh and spent medium and 405 from biofilm samples. Three significantly changing compounds that matched to high scoring library fragment patterns were chosen for validation. Conclusion: Evaluation of the results demonstrates that the Q Exactive GC is suitable for metabolomics analysis using a targeted/untargeted methodology. Many of the results were as expected: e.g. rapid consumption of glucose and fructose from the medium regardless of the cell type. Modulation of sugar-phosphate levels also suggest that the pentose phosphate pathway could be enhanced in the cells from co-cultured biofilms. Untargeted metabolomics results suggested significant production of cell-wall biosynthesis components and the consumption of non-proteinaceous amino-acids

Anterior Talofibular Ligament Repair With Suture Tape Augmentation

The most common injury sustained to the ankle ligaments is a result of inversion of the foot. This mechanism results in injury to the anterior talofibular ligament alone or in conjunction with the calcaneofibular ligament and posterior talofibular ligament. Patients experiencing recurrent ankle sprains despite nonoperative measures often require surgical management. Recent focus has been on augmentation procedures to improve the stability of a lateral ankle ligament repair by protecting it during the healing phase and supporting early mobilization. This article describes, with video illustration, anterior talofibular ligament repair with suture tape augmentation