Synthetic heparinoids labelled with 125I and 35S

The labelling of a water-soluble synthetic polyelectrolyte, having anticoagulant activity, has been studied. The polyelectrolyte is derived from cis-1,4-polyisoprene and contains N-sulfate and carboxylate groups. [125I]-Iodination of the polyelectrolyte, using the Chloramine-T method and an electrolytic method, resulted in a [125I]-labelled polyelectrolyte from which release of the label occurred. Resulfation of a partially desulfated polyelectrolyte with a [35S]-sulfur trioxide trimethylamine complex resulted in a [35S]-labelled polyelectrolyte which showed no release of the label

Atomistic simulations of the human proteasome inhibited by a covalent ligand

The proteasome is a large biomolecular complex responsible for protein degradation. It is under intense research due to its fundamental role in cellular homeostasis, and tremendous potential for medicinal applications. Recent data from X-ray crystallography and cryo-electron microscopy have suggested that there is a large-scale structural change upon binding of an inhibitor. We carried out atomistic molecular dynamics simulations of the native and inhibited proteasomes to understand the molecular details of the inhibition. Here we describe the technical details of the simulations and assess the quality of the trajectories obtained. The biochemical aspects of the proteasome are under further investigation and will be published elsewhere. This work was a part of the GCS-Prot project at the HLRS, run on the Cray XC40 supercomputing system

Timeline mapping in qualitative interviews: a study of resilience with marginalized groups

Growing interest in visual timeline methods signals a need for critical engagement. Drawing on critical emancipatory epistemologies in our study exploring resilience among marginalized groups, we investigate how the creation of visual timelines informs verbal semistructured interviewing. We consider both how experiences of drawing timelines and how the role of the timeline in interviews varied for South Asian immigrant women who experienced domestic violence, and street-involved youth who experienced prior or recent violent victimization. Here we focus on three overarching themes developed through analysis of timelines: (a) rapport building, (b) participants as navigators, and (c) therapeutic moments and positive closure. In the discussion, we engage with the potential of visual timelines to supplement and situate semistructured interviewing, and illustrate how the framing of research is central to whether that research maintains a critical emancipatory orientation

Effects of Oil and Gas Development on Mule Deer Populations in Western North Dakota and Eastern Montana

Oil and gas production are becoming a significant part of the economy and landscape of western North Dakota and eastern Montana.  Much of the areas being developed overlap with mule deer ranges.  Our ongoing research aims to identify and quantify the direct and indirect effects of oil and gas energy development on mule deer abundance, survival, recruitment, movements and resource selection.  Since February, 2013, we have deployed 240 GPS collars in three main areas of breaks habitat: 1) in North Dakota along the Little Missouri River; 2) the east side of the Yellowstone River; and 3) just south of Culbertson, MT. These collars are being used to collect spatial data about mule deer distributions and monitor survival across areas of low, medium, high energy development.  We will also use digitized aerial survey data to estimate abundance and recruitment across various levels of development. To date we have collared 99 adult females and 110 fawns, gathering more than 300,000 deer locations, conducted 39 lab necropsies on full and partial carcasses, and conducted biannual aerial surveys in North Dakota (2 years) and Montana (1 year). Our research will address potential impacts to mule deer populations, but will also provide mitigation strategies to help minimize disturbances from further development

A Modified Direct Power Control Strategy Allowing the Connection of Three-Phase Inverters to the Grid Through LCL Filters

Abstract — This paper proposes a novel approach to adapt the conventional Direct Power Control (DPC) for high power applications with a third order LCL filter. The strong resonance present in the LCL filter is damped with additional effort in the system control. The application of DPC to the control of threephase Voltage Source Inverter (VSI) connected to the grid through a LCL filter has not yet been considered. An active damping strategy for the LCL filter together with harmonic rejection control is proposed over the conventional DPC. The steady state as well as the dynamic performance of the proposed system is presented by means of the simulation results and compared with the conventional approach

Use of genetic algorithms with multivariate regression for determination of gelatine in historic papers based on FT-IR and NIR spectral data

Quantitative non-destructive analysis of individual constituents of historic rag paper is crucial for its effective preservation. In this work, we examine the potentials of mid- and near-infrared spectroscopy, however, in order to fully utilise the selectivity inherent to spectroscopic multivariate measurements, genetic algorithms were used to select spectral data derived from information-rich FT-IR or UV-vis-NIR measurements to build multivariate calibration models based on partial least squares regression, relating spectra to gelatine content in paper. A selective but laborious chromatographic method for the quantification of hydroxyproline (HYP) has been developed to provide the reference data on gelatine content. We used 9-fluorenylmethyl chloroformate (FMOC) to derivatise HYP, which was subsequently determined using reverse-phase liquid chromatographic separation and fluorimetric detection. In this process, the sample is consumed, which is why the method can only be used as a reference method.The sampling flexibility afforded by small-size field-portable spectroscopic instrumentation combined with chemometric data analysis, represents an attractive addition to existing analytical techniques for cultural heritage materials. (C) 2010 Elsevier B.V. All rights reserved

Conforming finite element methods for the clamped plate problem

Finite element methods for solving biharmonic boundary value problems are considered. The particular problem discussed is that of a clamped thin plate. This problem is reformulated in a weak, form in the Sobolev space Techniques for setting up conforming trial Functions are utilized in a Galerkin technique to produce finite element solutions. The shortcomings of various trial function formulations are discussed, and a macro—element approach to local mesh refinement using rectangular elements is given

Importance of Bacillithiol in the Oxidative Stress Response of Staphylococcus aureus

In Staphylococcus aureus, the low-molecular-weight thiol called bacillithiol (BSH), together with cognate S-transferases, is believed to be the counterpart to the glutathione system of other organisms. To explore the physiological role of BSH in S. aureus, we constructed mutants with the deletion of bshA (sa1291), which encodes the glycosyltransferase that catalyzes the first step of BSH biosynthesis, and fosB (sa2124), which encodes a BSH-S-transferase that confers fosfomycin resistance, in several S. aureus strains, including clinical isolates. Mutation of fosB or bshA caused a 16- to 60-fold reduction in fosfomycin resistance in these S. aureus strains. High-pressure liquid chromatography analysis, which quantified thiol extracts, revealed some variability in the amounts of BSH present across S. aureus strains. Deletion of fosB led to a decrease in BSH levels. The fosB and bshA mutants of strain COL and a USA300 isolate, upon further characterization, were found to be sensitive to H2O2 and exhibited decreased NADPH levels compared with those in the isogenic parents. Microarray analyses of COL and the isogenic bshA mutant revealed increased expression of genes involved in staphyloxanthin synthesis in the bshA mutant relative to that in COL under thiol stress conditions. However, the bshA mutant of COL demonstrated decreased survival compared to that of the parent in human whole-blood survival assays; likewise, the naturally BSH-deficient strain SH1000 survived less well than its BSH-producing isogenic counterpart. Thus, the survival of S. aureus under oxidative stress is facilitated by BSH, possibly via a FosB-mediated mechanism, independently of its capability to produce staphyloxanthin

Folding of VemP into translation-arresting secondary structure is driven by the ribosome exit tunnel

The ribosome is a fundamental biomolecular complex that synthesizes proteins in cells. Nascent proteins emerge from the ribosome through a tunnel, where they may interact with the tunnel walls or small molecules such as antibiotics. These interactions can cause translational arrest with notable physiological consequences. Here, we studied the arrest caused by the regulatory peptide VemP, which is known to form alpha-helices inside the ribosome tunnel near the peptidyl transferase center under specific conditions. We used all-atom molecular dynamics simulations of the entire ribosome and circular dichroism spectroscopy to study the driving forces of helix formation and how VemP causes the translational arrest. To that aim, we compared VemP dynamics in the ribosome tunnel with its dynamics in solution. We show that the VemP peptide has a low helical propensity in water and that the propensity is higher in mixtures of water and trifluorethanol. We propose that helix formation within the ribosome is driven by the interactions of VemP with the tunnel and that a part of VemP acts as an anchor. This anchor might slow down VemP progression through the tunnel enabling alpha-helix formation, which causes the elongation arrest