Comparison of Extensive Protein Fractionation and Repetitive LC-MS/MS Analyses on Depth of Analysis for Complex Proteomes

In-depth, reproducible coverage of complex proteomes is challenging because the complexity of tryptic digests subjected to LC-MS/MS analysis frequently exceeds mass spectrometer analytical capacity, which results in undersampling of data. In this study, we used cancer cell lysates to systematically compare the commonly used GeLC-MS/MS (1-D protein + 1-D peptide separation) method using four repetitive injections (2-D/repetitive) with a 3-D method that included solution isoelectric focusing and involved an equal number of LC-MS/MS runs. The 3-D method detected substantially more unique peptides and proteins, including higher numbers of unique peptides from low-abundance proteins, demonstrating that additional fractionation at the protein level is more effective than repetitive analyses at overcoming LC-MS/MS undersampling. Importantly, more than 90 % of the 2-D/repetitive protein identifications were found in the 3-D method data in a direct protein level comparison, and the reproducibility between data sets increased to greater than 96 % when factors such as database redundancy and use of rigid scoring thresholds were considered. Hence, high reproducibility of complex proteomes, such as human cancer cell lysates, readily can be achieved when using multidimensional separation methods with good depth of analysis

Process and impact evaluation of the Greater Christchurch Urban Development Strategy Health Impact Assessment

<p>Abstract</p> <p>Background</p> <p>despite health impact assessment (HIA) being increasingly widely used internationally, fundamental questions about its impact on decision-making, implementation and practices remain. In 2005 a collaboration between public health and local government authorities performed an HIA on the Christchurch Urban Development Strategy Options paper in New Zealand. The findings of this were incorporated into the Greater Christchurch Urban Development Strategy;</p> <p>Methods</p> <p>using multiple qualitative methodologies including key informant interviews, focus groups and questionnaires, this study performs process and impact evaluations of the Christchurch HIA including evaluation of costs and resource use;</p> <p>Results</p> <p>the evaluation found that the HIA had demonstrable direct impacts on planning and implementation of the final Urban Development Strategy as well as indirect impacts on understandings and ways of working within and between organisations. It also points out future directions and ways of working in this successful collaboration between public health and local government authorities. It summarises the modest resource use and discusses the important role HIA can play in urban planning with intersectoral collaboration and enhanced relationships as both catalysts and outcomes of the HIA process;</p> <p>Conclusion</p> <p>as one of the few evaluations of HIA that have been published to date, this paper makes a substantial contribution to the literature on the impact, utility and effectiveness of HIA.</p

WSES guidelines for management of Clostridium difficile infection in surgical patients

In the last two decades there have been dramatic changes in the epidemiology of Clostridium difficile infection (CDI), with increases in incidence and severity of disease in many countries worldwide. The incidence of CDI has also increased in surgical patients. Optimization of management of C difficile, has therefore become increasingly urgent. An international multidisciplinary panel of experts prepared evidenced-based World Society of Emergency Surgery (WSES) guidelines for management of CDI in surgical patients

WSES guidelines for management of Clostridium difficile infection in surgical patients

In the last two decades there have been dramatic changes in the epidemiology of Clostridium difficile infection (CDI), with increases in incidence and severity of disease in many countries worldwide. The incidence of CDI has also increased in surgical patients. Optimization of management of C difficile, has therefore become increasingly urgent. An international multidisciplinary panel of experts prepared evidenced-based World Society of Emergency Surgery (WSES) guidelines for management of CDI in surgical patients.Peer reviewe

An Isolated Stellar-Mass Black Hole Detected Through Astrometric Microlensing

We report the first unambiguous detection and mass measurement of an isolated stellar-mass black hole (BH). We used the Hubble Space Telescope (HST) to carry out precise astrometry of the source star of the long-duration (t_E ~ 270 days), high-magnification microlensing event MOA-2011-BLG-191/OGLE-2011-BLG-0462, in the direction of the Galactic bulge. HST imaging, conducted at eight epochs over an interval of six years, reveals a clear relativistic astrometric deflection of the background star's apparent position. Ground-based photometry shows a parallactic signature of the effect of the Earth's motion on the microlensing light curve. Combining the HST astrometry with the ground-based light curve and the derived parallax, we obtain a lens mass of 7.1 +/- 1.3 M_Sun and a distance of 1.58 +/- 0.18 kpc. We show that the lens emits no detectable light, which, along with having a mass higher than is possible for a white dwarf or neutron star, confirms its BH nature. Our analysis also provides an absolute proper motion for the BH. The proper motion is offset from the mean motion of Galactic-disk stars at similar distances by an amount corresponding to a transverse space velocity of ~45 km/s, suggesting that the BH received a modest natal 'kick' from its supernova explosion. Previous mass determinations for stellar-mass BHs have come from radial-velocity measurements of Galactic X-ray binaries, and from gravitational radiation emitted by merging BHs in binary systems in external galaxies. Our mass measurement is the first ever for an isolated stellar-mass BH using any technique

Interviewing adults with learning diasabilities about sexual abuse

DraftAvailable from British Library Document Supply Centre-DSC:q96/07819 / BLDSC - British Library Document Supply CentreSIGLEGBUnited Kingdo

Metabolism of bovine parathyroid hormone. Immunological and biological characteristics of fragments generated by liver perfusion.

The metabolism of bovine parathyroid hormone (PTH) by the perfused rat liver was studied. Labeled hormone, with or without cold hormone, was infused into the circulating perfusion medium containing various calcium concentrations. Pefusate samples at various time periods after the introduction of PTH into the system were chromatographed on Bio-gel P-10; radioactivity and/or immunoreactivity were measured in eluted fractions. Before the perfusion, all immuno- and radioactivity eluted in a single peak, with an apparent mol wt of 9,500 (peak I). After perfusion for 15 min, two other peaks with approximate mol wt of 7,000 (peak II) and 3,500 (peak III) were discernible. Peak I contained both NH2-terminal and COOH-terminal immunoreactivity and was biologically active at all time periods tested. The relative contribution of NH2-terminal and COOH-terminal immunoreactivity to the total immunoreactivity remained constant in this peak throughout the perfusion. In every respect, peak I had the characteristics of intact hormone. At all times, peak II consisted of only COOH-terminal immunoreactivity and was biologically inactive. At early time periods, peak III contained predominantly NH2-terminal immunoreactivity and was biologically active. With time, the relative contribution of NH2-terminal immunoreactivity decreased strikingly while that of COOH-terminal immunoreactivity increased. The three peaks identified in these experiments were analogous in size, biological activity, and immunological characteristics to those we have previously described for fractionated human hyperparathyroid serum. The rate of metabolism of PTH appeared to be regulated by the calcium concentration in the medium. At a high concentration of calcium (greater than 11 mg/100 ml), PTH metabolism was greatly retarded. At a low concentration of calcium (smaller than 5 mg/100 ml), the rate of metabolism was greatly increased. The physiological significance of our observations on the metabolism of PTH by isolated perfused rat liver is not known. However, since such metabolism results in a biologically active fragment, it is suggested that metabolism of intact hormone may be required before full biological expression is possible