Optimization Of Detergent-Mediated Reconstitution Of Influenza A M2 Protein Into Proteoliposomes

We report the optimization of detergent-mediated reconstitution of an integral membrane-bound protein, full-length influenza M2 protein, by direct insertion into detergent-saturated liposomes. Detergent-mediated reconstitution is an important method for preparing proteoliposomes for studying membrane proteins, and must be optimized for each combination of protein and membrane constituents used. The purpose of the reconstitution was to prepare samples for site-directed spin-labeling electron paramagnetic resonance (SDSL-EPR) studies. Our goals in optimizing the protocol were to minimize the amount of detergent used, reduce overall proteoliposome preparation time, and confirm the removal of all detergent. The liposomes were comprised of (1-palmitoyl-2-oleyl-sn-glycero-phosphocholine (POPC) and 1-palmitoyl-2-oleyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (POPG), and the detergent octylglucoside (OG) was used for reconstitution. Rigorous physical characterization was applied to optimize each step of the reconstitution process. We used dynamic light scattering (DLS) to determine the amount of OG needed to saturate the preformed liposomes. During detergent removal by absorption with Bio-Beads, we quantified the detergent concentration by means of a colorimetric assay, thereby determining the number of Bio-Bead additions needed to remove all detergent from the final proteoliposomes. We found that the overnight Bio-Bead incubation used in previously published protocols can be omitted, reducing the time needed for reconstitution. We also monitored the size distribution of the proteoliposomes with DLS, confirming that the size distribution remains essentially constant throughout the reconstitution process

Toward a Dependability Case Language and Workflow for a Radiation Therapy System

We present a near-future research agenda for bringing a suite of modern programming-languages verification tools - specifically interactive theorem proving, solver-aided languages, and formally defined domain-specific languages - to the development of a specific safety-critical system, a radiotherapy medical device. We sketch how we believe recent programming-languages research advances can merge with existing best practices for safety-critical systems to increase system assurance and developer productivity. We motivate hypotheses central to our agenda: That we should start with a single specific system and that we need to integrate a variety of complementary verification and synthesis tools into system development

A gravitational lensing explanation for the excess of strong Mg-II absorbers in GRB afterglow spectra

GRB afterglows offer a probe of the intergalactic medium out to high redshift which complements observations along more abundant quasar lines-of-sight. Although both quasars and GRB afterglows should provide a-priori random sight-lines through the intervening IGM, it has been observed that strong Mg-II absorbers are twice as likely to be found along sight-lines toward GRBs. Several proposals to reconcile this discrepancy have been put forward, but none has been found sufficient to explain the magnitude of the effect. In this paper we estimate the effect of gravitational lensing by galaxies and their surrounding mass distributions on the statistics of Mg-II absorption. We find that the multi-band magnification bias could be very strong in the spectroscopic GRB afterglow population and that gravitational lensing can explain the discrepancy in density of absorbers, for plausibly steep luminosity functions. The model makes the prediction that approximately 20%-60% of the spectroscopic afterglow sample (i.e. ~ 5-15 of 26 sources) would have been multiply imaged, and hence result in repeating bursts. We show that despite this large lensing fraction it is likely that none would yet have been identified by chance owing to the finite sky coverage of GRB searches. We predict that continued optical monitoring of the bright GRB afterglow locations in the months and years following the initial decay would lead to identification of lensed GRB afterglows. A confirmation of the lensing hypothesis would allow us to constrain the GRB luminosity function down to otherwise inaccessibly faint levels, with potential consequences for GRB models.Comment: 8 pages, 3 figures. Submitted to MNRAS

Medically unexplained symptoms and attachment theory: The BodyMind Approach

© 2019 Payne and Brooks. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.This article discusses how The BodyMind Approach ® (TBMA) addresses insecure attachment styles in medically unexplained symptoms (MUS). Insecure attachment styles are associated with adverse childhood experiences (ACEs) and MUS (Adshead and Guthrie, 2015) and affect sufferers' capacity to self-manage. The article goes on to make a new hypothesis to account for TBMA's effectiveness (Payne and Brooks, 2017), that is, it addresses insecure attachment styles, which may be present in some MUS sufferers, leading to their capacity to self-manage. Three insecure attachment styles (dismissive, pre-occupied and fearful) associated with MUS are discussed. TBMA is described and explanations provided of how TBMA has been specifically designed to support people's insecure attachment styles. Three key concepts to support insecure attachment styles involved in the content of TBMA are identified and debated: (a) emotional regulation; (b) safety; and (c) bodymindfulness. There is a rationale for the design of TBMA as opposed to psychological interventions for this population. The programme's structure, facilitation and content, takes account of the three insecure attachment styles above. Examples of how TBMA works with their specific characteristics are presented. TBMA has been tested and found to be effective during delivery in the United Kingdom National Health Service (NHS). Improved self-management has potential to reduce costs for the NHS and in General Practitioner time and resources.Peer reviewe