Comprehensive mapping of O-GlcNAc modification sites using a chemically cleavable tag

The post-translational modification of serine or threonine residues of proteins with a single N-acetylglucosamine monosaccharide (O-GlcNAcylation) is essential for cell survival and function. However, relatively few O-GlcNAc modification sites have been mapped due to the difficulty of enriching and detecting O-GlcNAcylated peptides from complex samples. Here we describe an improved approach to quantitatively label and enrich O-GlcNAcylated proteins for site identification. Chemoenzymatic labelling followed by copper(I)-catalysed azide–alkyne cycloaddition (CuAAC) installs a new mass spectrometry (MS)-compatible linker designed for facile purification of O-GlcNAcylated proteins from cell lysates. The linker also allows subsequent quantitative release of O-GlcNAcylated proteins for downstream MS analysis. We validate the approach by unambiguously identifying several established O-GlcNAc sites on the proteins α-crystallin and O-GlcNAc transferase (OGT), as well as discovering new, previously unreported sites on OGT. Notably, these novel sites on OGT lie in key functional domains of the protein, underscoring how this site identification method may reveal important biological insights into protein activity and regulation

Real-time RMS active damping augmentation: Heavy and very light payload evaluations

Controls-Structures Integration Technology has been applied to the Space Shuttle Remote Manipulator System (RMS) to improve on-orbit performance. The objective was to actively damp undesired oscillatory motions of the RMS following routine payload maneuvering and Shuttle attitude control thruster firings. Simulation of active damping was conducted in the real-time, man-in-the-loop Systems Engineering Simulator at NASA's Johnson Space Center. The simulator was used to obtain qualitative and quantitative data on active damping performance from astronaut operators. Using a simulated three-axis accelerometer mounted on the RMS, 'sensed' vibration motions were used to generate joint motor commands that reduced the unwanted oscillations. Active damping of the RMS with heavy and light attached payloads was demonstrated in this study. Five astronaut operators examined the performance of active damping following operator commanded RMS maneuvers and Shuttle thruster firings. Noticeable improvements in the damping response of the RMS with the heavy, Hubble Space Telescope payload and the very light, astronaut in Manipulator Foot Restraint payload were observed. The potential of active damping to aid in precisely maneuvering payloads was deemed significant

Effect of Opioid vs Nonopioid Medications on Pain-Related Function in Patients With Chronic Back Pain or Hip or Knee Osteoarthritis Pain

Importance: Limited evidence is available regarding long-term outcomes of opioids compared with nonopioid medications for chronic pain. Objective: To compare opioid vs nonopioid medications over 12 months on pain-related function, pain intensity, and adverse effects. Design, Setting, and Participants: Pragmatic, 12-month, randomized trial with masked outcome assessment. Patients were recruited from Veterans Affairs primary care clinics from June 2013 through December 2015; follow-up was completed December 2016. Eligible patients had moderate to severe chronic back pain or hip or knee osteoarthritis pain despite analgesic use. Of 265 patients enrolled, 25 withdrew prior to randomization and 240 were randomized. Interventions: Both interventions (opioid and nonopioid medication therapy) followed a treat-to-target strategy aiming for improved pain and function. Each intervention had its own prescribing strategy that included multiple medication options in 3 steps. In the opioid group, the first step was immediate-release morphine, oxycodone, or hydrocodone/acetaminophen. For the nonopioid group, the first step was acetaminophen (paracetamol) or a nonsteroidal anti-inflammatory drug. Medications were changed, added, or adjusted within the assigned treatment group according to individual patient response. Main Outcomes and Measures: The primary outcome was pain-related function (Brief Pain Inventory [BPI] interference scale) over 12 months and the main secondary outcome was pain intensity (BPI severity scale). For both BPI scales (range, 0-10; higher scores = worse function or pain intensity), a 1-point improvement was clinically important. The primary adverse outcome was medication-related symptoms (patient-reported checklist; range, 0-19). Results: Among 240 randomized patients (mean age, 58.3 years; women, 32 [13.0%]), 234 (97.5%) completed the trial. Groups did not significantly differ on pain-related function over 12 months (overall P = .58); mean 12-month BPI interference was 3.4 for the opioid group and 3.3 for the nonopioid group (difference, 0.1 [95% CI, -0.5 to 0.7]). Pain intensity was significantly better in the nonopioid group over 12 months (overall P = .03); mean 12-month BPI severity was 4.0 for the opioid group and 3.5 for the nonopioid group (difference, 0.5 [95% CI, 0.0 to 1.0]). Adverse medication-related symptoms were significantly more common in the opioid group over 12 months (overall P = .03); mean medication-related symptoms at 12 months were 1.8 in the opioid group and 0.9 in the nonopioid group (difference, 0.9 [95% CI, 0.3 to 1.5]). Conclusions and Relevance: Treatment with opioids was not superior to treatment with nonopioid medications for improving pain-related function over 12 months. Results do not support initiation of opioid therapy for moderate to severe chronic back pain or hip or knee osteoarthritis pain

Loss of O-GlcNAc glycosylation in forebrain excitatory neurons induces neurodegeneration

O-GlcNAc glycosylation (or O-GlcNAcylation) is a dynamic, inducible posttranslational modification found on proteins associated with neurodegenerative diseases such as α-synuclein, amyloid precursor protein, and tau. Deletion of the O-GlcNAc transferase (ogt) gene responsible for the modification causes early postnatal lethality in mice, complicating efforts to study O-GlcNAcylation in mature neurons and to understand its roles in disease. Here, we report that forebrain-specific loss of OGT in adult mice leads to progressive neurodegeneration, including widespread neuronal cell death, neuroinflammation, increased production of hyperphosphorylated tau and amyloidogenic Aβ-peptides, and memory deficits. Furthermore, we show that human cortical brain tissue from Alzheimer’s disease patients has significantly reduced levels of OGT protein expression compared with cortical tissue from control individuals. Together, these studies indicate that O-GlcNAcylation regulates pathways critical for the maintenance of neuronal health and suggest that dysfunctional O-GlcNAc signaling may be an important contributor to neurodegenerative diseases

From theory to 'measurement' in complex interventions: methodological lessons from the development of an e-health normalisation instrument

<b>Background</b> Although empirical and theoretical understanding of processes of implementation in health care is advancing, translation of theory into structured measures that capture the complex interplay between interventions, individuals and context remain limited. This paper aimed to (1) describe the process and outcome of a project to develop a theory-based instrument for measuring implementation processes relating to e-health interventions; and (2) identify key issues and methodological challenges for advancing work in this field.<p></p> <b>Methods</b> A 30-item instrument (Technology Adoption Readiness Scale (TARS)) for measuring normalisation processes in the context of e-health service interventions was developed on the basis on Normalization Process Theory (NPT). NPT focuses on how new practices become routinely embedded within social contexts. The instrument was pre-tested in two health care settings in which e-health (electronic facilitation of healthcare decision-making and practice) was used by health care professionals.<p></p> <b>Results</b> The developed instrument was pre-tested in two professional samples (N = 46; N = 231). Ratings of items representing normalisation 'processes' were significantly related to staff members' perceptions of whether or not e-health had become 'routine'. Key methodological challenges are discussed in relation to: translating multi-component theoretical constructs into simple questions; developing and choosing appropriate outcome measures; conducting multiple-stakeholder assessments; instrument and question framing; and more general issues for instrument development in practice contexts.<p></p> <b>Conclusions</b> To develop theory-derived measures of implementation process for progressing research in this field, four key recommendations are made relating to (1) greater attention to underlying theoretical assumptions and extent of translation work required; (2) the need for appropriate but flexible approaches to outcomes measurement; (3) representation of multiple perspectives and collaborative nature of work; and (4) emphasis on generic measurement approaches that can be flexibly tailored to particular contexts of study

Comparative genomics of Toll-like receptor signalling in five species

<p>Abstract</p> <p>Background</p> <p>Over the last decade, several studies have identified quantitative trait loci (QTL) affecting variation of immune related traits in mammals. Recent studies in humans and mice suggest that part of this variation may be caused by polymorphisms in genes involved in Toll-like receptor (TLR) signalling. In this project, we used a comparative approach to investigate the importance of TLR-related genes in comparison with other immunologically relevant genes for resistance traits in five species by associating their genomic location with previously published immune-related QTL regions.</p> <p>Results</p> <p>We report the genomic localisation of <it>TLR1-10 </it>and ten associated signalling molecules in sheep and pig using <it>in-silico </it>and/or radiation hybrid (RH) mapping techniques and compare their positions with their annotated homologues in the human, cattle and mouse whole genome sequences. We also report medium-density RH maps for porcine chromosomes 8 and 13. A comparative analysis of the positions of previously published relevant QTLs allowed the identification of homologous regions that are associated with similar health traits in several species and which contain TLR related and other immunologically relevant genes. Additional evidence was gathered by examining relevant gene expression and association studies.</p> <p>Conclusion</p> <p>This comparative genomic approach identified eight genes as potentially causative genes for variations of health related traits. These include susceptibility to clinical mastitis in dairy cattle, general disease resistance in sheep, cattle, humans and mice, and tolerance to protozoan infection in cattle and mice. Four TLR-related genes (<it>TLR1</it>, <it>6</it>, <it>MyD88</it>, <it>IRF3</it>) appear to be the most likely candidate genes underlying QTL regions which control the resistance to the same or similar pathogens in several species. Further studies are required to investigate the potential role of polymorphisms within these genes.</p

Abundances in Turn-off Stars in the Old, Metal-Rich Open Cluster, NGC 6791

Open clusters have long been used to illuminate both stellar evolution and Galactic evolution. The oldest clusters, though rather rare, can reveal the chemical and nucleosynthetic processes early in the history of the Galaxy. We have studied two turn-off stars in the old, metal-rich open cluster, NGC 6791. The Keck + HIRES spectra have a resolution of 45,000 and signal-to-noise ratios of 40 per pixel. We confirm the high value for [Fe/H] finding +0.30 $\pm$0.08, in agreement with earlier results from evolved stars in other parts of the HR diagram. We have also determined abundances for Na, Si, Ca, Ti, Cr, Ni, Y and Ba. These are compared to a sample of old, metal-rich field stars. With the probable exception of enhanced Ni in the cluster stars, the field and cluster stars show similar abundances of the elements. Model predictions show that the Ni enhancement could result from enrichment of the pre-cluster gas by SN Ia. Orbital evidence indicates that NGC 6791 could have originated near the inner regions of the Galaxy where the metallicity is generally higher than it is in the disk or halo. Subsequent perturbations and migrations may have resulted in its current heliocentric distance of 4 kpc and 1 kpc above the Galactic plane.Comment: 10 pages, 4 figures, 7 tables accepted by The Astronomical Journal for June, 200

Using interpretative phenomenological analysis to inform physiotherapy practice: An introduction with reference to the lived experience of cerebellar ataxia

The attached file is a pre-published version of the full and final paper which can be found at the link below.This article has been made available through the Brunel Open Access Publishing Fund.Qualitative research methods that focus on the lived experience of people with health conditions are relatively underutilised in physiotherapy research. This article aims to introduce interpretative phenomenological analysis (IPA), a research methodology oriented toward exploring and understanding the experience of a particular phenomenon (e.g., living with spinal cord injury or chronic pain, or being the carer of someone with a particular health condition). Researchers using IPA try to find out how people make sense of their experiences and the meanings they attach to them. The findings from IPA research are highly nuanced and offer a fine grained understanding that can be used to contextualise existing quantitative research, to inform understanding of novel or underresearched topics or, in their own right, to provoke a reappraisal of what is considered known about a specified phenomenon. We advocate IPA as a useful and accessible approach to qualitative research that can be used in the clinical setting to inform physiotherapy practice and the development of services from the perspective of individuals with particular health conditions.This article is available through the Brunel Open Access Publishing Fund

Data Generated during the 2018 LAPSE-RATE Campaign: An Introduction and Overview

Unmanned aircraft systems (UASs) offer innovative capabilities for providing new perspectives on the atmosphere, and therefore atmospheric scientists are rapidly expanding their use, particularly for studying the planetary boundary layer. In support of this expansion, from 14 to 20 July 2018 the International Society for Atmospheric Research using Remotely piloted Aircraft (ISARRA) hosted a community flight week, dubbed the Lower Atmospheric Profiling Studies at Elevation – a Remotely-piloted Aircraft Team Experiment (LAPSE-RATE; de Boer et al., 2020a). This field campaign spanned a 1-week deployment to Colorado\u27s San Luis Valley, involving over 100 students, scientists, engineers, pilots, and outreach coordinators. These groups conducted intensive field operations using unmanned aircraft and ground-based assets to develop comprehensive datasets spanning a variety of scientific objectives, including a total of nearly 1300 research flights totaling over 250 flight hours. This article introduces this campaign and lays the groundwork for a special issue on the LAPSE-RATE project. The remainder of the special issue provides detailed overviews of the datasets collected and the platforms used to collect them. All of the datasets covered by this special issue have been uploaded to a LAPSE-RATE community set up at the Zenodo data archive (https://zenodo.org/communities/lapse-rate/, last access: 3 December 2020)