Examining mechanisms underlying the selective vulnerability of motor units in a mouse model of Spinal Muscular Atrophy

Spinal Muscular Atrophy (SMA) is a childhood form of motor neuron disease that causes a progressive paralysis that, in its most severe form, results in death before two years of age. There is currently no cure or treatment for SMA. SMA is caused by a reduction in levels of Survival Motor Neuron (SMN) protein, which results in the degeneration of lower motor neurons. This degeneration is first observed at the neuromuscular junction (NMJ), where pre-synaptic nerve terminals belonging to the motor neuron become dysfunctional and degenerate during the early stages of disease. Several previous studies have shown that the some populations of motor neurons appear to have a resistance to SMA pathology, while other neighbouring populations are vulnerable. In this study, we attempted to elucidate the cause of this vulnerability spectrum. Initially, we characterised the relative vulnerability of ten different motor unit pools in an established mouse model of severe SMA and attempted to correlate these vulnerabilities with quantified aspects of motor unit morphology. From this study, no significant correlation could be found with any aspect of motor unit morphology examined, suggesting that morphological parameters of motor neurons do no influence their relative susceptibility. We then attempted to identify changes in basal gene expression between protected and vulnerable pools of motor units using microarray analysis. Motor unit pools were labelled using a retrograde tracer injected into muscles that had previously been identified as having highly vulnerable or resistant motor units. Labelled motor neuron cell bodies were then isolated from the spinal cord using laser capture micro-dissection and RNA was extracted for microarray analysis. From this study, we identified several molecular pathways and individual genes whose expression levels compared the gene expression profiles of vulnerable and resistant motor units. Thus, molecular differences between motor neuron pools likely underlie their relative vulnerability to degeneration in SMA. Lastly, we attempted to identify a novel peptide that could be used to label synapses, including neuromuscular junctions, in vivo. This would allow us to non-invasively visualise degenerating NMJs and other synapses in human patients without the need for a biopsy. Such a tool would be extremely valuable in assessing the effectiveness of drug trials, both in human patients and animal models, and may also contribute to earlier diagnosis of motor neuron disorders. To identify a potentially suitable peptide, we used a phage display library and panned for peptides that specifically bound to the outer surface of synapses using synaptosome preparations. From this panning we successfully enriched two peptides, the sequences of which were used to manufacture fluorescently tagged peptides

“About sixty per cent I want to do it” : Health researchers’ attitudes to, and experiences of, patient and public involvement (PPI)—A qualitative interview study

This research was funded through a personal fellowship awarded to Louise Locock by the NIHR Oxford Biomedical Research Centre. Anne‐Marie Boylan received support from the NIHR Collaboration for Leadership in Applied Research and Care Oxford. Sophie Staniszewska is part‐funded by the NIHR Collaboration for Leadership in Applied Research and Care West Midlands. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. ACKNOWLEDGEMENTS We are grateful to the participants for sharing their experiences to inform this research. This research was supported by a panel comprising patients, members of the public, AND health, clinical and medical researchers. We are grateful for their support.Peer reviewedPublisher PD

Enhanced Tissue Integration During Cartilage RepairIn VitroCan Be Achieved by Inhibiting Chondrocyte Death at the Wound Edge

Objective: Experimental wounding of articular cartilage results in cell death at the lesion edge. The objective of this study was to investigate whether inhibition of this cell death results in enhanced integrative cartilage repair. Methods: Bovine articular cartilage discs (6mm) were incubated in media containing inhibitors of necrosis (Necrostatin-1, Nec-1) or apoptosis (Z-VAD-FMK, ZVF) before cutting a 3mm inner core. This core was left in situ to create disc/ring composites, cultured for up to 6 weeks with the inhibitors, and analyzed for cell death, sulfated glycosaminoglycan release, and tissue integration. Results: Creating the disc/ring composites resulted in a significant increase in necrosis. ZVF significantly reduced necrosis and apoptosis at the wound edge. Nec-1 reduced necrosis. Both inhibitors reduced the level of wound-induced sulfated glycosaminoglycan loss. Toluidine blue staining and electron microscopy of cartilage revealed significant integration of the wound edges in disc/ring composites treated with ZVF. Nec-1 improved integration, but to a lesser extent. Push-out testing revealed that ZVF increased adhesive strength compared to control composites. Conclusions: This study shows that treatment of articular cartilage with cell death inhibitors during wound repair increases the number of viable cells at the wound edge, prevents matrix loss, and results in a significant improvement in cartilage-cartilage integration

Gut mucosal microbiome signatures of colorectal cancer differ according to BMI status

ACKNOWLEDGMENTS We gratefully acknowledge the NHS Grampian Biorepository for providing access to CRC patient samples and data. We thank members of the GI Research Team for discussions and advice. The authors thank Brennan Martin and the Centre for Genome Enabled Biology and Medicine for useful discussions.Peer reviewedPublisher PD

Lost in the Process?: The impact of devolution on abortion law in the United Kingdom

Using the case study of abortion policy across the United Kingdom, this article takes a feminist institutionalist approach to advance our understanding of state architecture and party competition within decentralised political systems. Despite increasing divergences across the United Kingdom in relation to abortion policy, contemporary debates around abortion access have rarely become politicised. Moreover, as this article demonstrates, when they have, the subject has been framed by politicians as a constitutional matter, relating to legislative competencies, rather than considered in terms of women’s rights. This framing, we argue, is linked to the specific constitutional arrangements of the post-devolution UK and the political strategies of the parties operating within them. Drawing upon parliamentary debates and interviews with political representatives to map the circumstances driving changes to abortion policy in the United Kingdom, this article introduces important comparative lessons for other cases of political decentralisation on the discussions and policies concerning women’s right

Nationwide Stepwise Emergence and Evolution of Multidrug-Resistant Campylobacter jejuni Sequence Type 5136, United Kingdom

We thank Food Standards Scotland (Contracts S14054, FSS00017) and the Scottish Government’s Rural and Environment Science and Analytical Services (RG13588-10) for funding this work. Part of this work was presented at the Campylobacter, Helicobacter and Related Organisms (CHRO) 2017 Congress, Nantes, France.Peer reviewedPublisher PD

Iterative focused screening with biological fingerprints identifies selective Asc-1 inhibitors distinct from traditional high throughput screening

N-methyl-d-aspartate receptors (NMDARs) mediate glutamatergic signaling that is critical to cognitive processes in the central nervous system, and NMDAR hypofunction is thought to contribute to cognitive impairment observed in both schizophrenia and Alzheimer’s disease. One approach to enhance the function of NMDAR is to increase the concentration of an NMDAR coagonist, such as glycine or d-serine, in the synaptic cleft. Inhibition of alanine–serine–cysteine transporter-1 (Asc-1), the primary transporter of d-serine, is attractive because the transporter is localized to neurons in brain regions critical to cognitive function, including the hippocampus and cortical layers III and IV, and is colocalized with d-serine and NMDARs. To identify novel Asc-1 inhibitors, two different screening approaches were performed with whole-cell amino acid uptake in heterologous cells stably expressing human Asc-1: (1) a high-throughput screen (HTS) of 3 M compounds measuring 35S l-cysteine uptake into cells attached to scintillation proximity assay beads in a 1536 well format and (2) an iterative focused screen (IFS) of a 45 000 compound diversity set using a 3H d-serine uptake assay with a liquid scintillation plate reader in a 384 well format. Critically important for both screening approaches was the implementation of counter screens to remove nonspecific inhibitors of radioactive amino acid uptake. Furthermore, a 15 000 compound expansion step incorporating both on- and off-target data into chemical and biological fingerprint-based models for selection of additional hits enabled the identification of novel Asc-1-selective chemical matter from the IFS that was not identified in the full-collection HTS

Art Now: An inquiry into the state of art and design teaching in early years foundation stage, primary and secondary education

The Art Now Inquiry explores the current state of art and design education across the four nations; however, the focus is primarily on England where there was more survey data to draw on. It was commissioned by the All-Party Parliamentary Group for Art, Craft and Design in Education in response to concerns about the reduction in opportunities for children and young people to access high-quality art and design education. The Inquiry spans early years, primary and secondary phases of schooling. The Inquiry ran between Spring 2020 and Spring 2023 and this report includes a rapid evidence review of the benefits of art and design education, a literature review of art teaching and teacher education, a national survey of 1,860 art and design teachers and testimonies from two APPG evidence sessions. Drawing on each of these sources, the Art Now Inquiry report makes the case for art and design education, and the critical importance of investing in a diverse subject-specialist workforce. It starts with an examination of teacher education in art and design which is essential for equipping teachers with the necessary skills, knowledge, and confidence to deliver the curriculum. The Inquiry goes behind the numbers to explore the working conditions, wellbeing and career intentions of art and design teachers. The findings provide a health check on the training and retention of art and design teachers, and highlight the time and resources needed to support access to high-quality provision of art, craft and design education. Ofsted defines a high-quality curriculum in art and design as one that provides the conditions for pupils to develop a love of the subject that is both intellectually challenging and creatively demanding

Morphological characteristics of motor neurons do not determine their relative susceptibility to degeneration in a mouse model of severe spinal muscular atrophy

Spinal muscular atrophy (SMA) is a leading genetic cause of infant mortality, resulting primarily from the degeneration and loss of lower motor neurons. Studies using mouse models of SMA have revealed widespread heterogeneity in the susceptibility of individual motor neurons to neurodegeneration, but the underlying reasons remain unclear. Data from related motor neuron diseases, such as amyotrophic lateral sclerosis (ALS), suggest that morphological properties of motor neurons may regulate susceptibility: in ALS larger motor units innervating fast-twitch muscles degenerate first. We therefore set out to determine whether intrinsic morphological characteristics of motor neurons influenced their relative vulnerability to SMA. Motor neuron vulnerability was mapped across 10 muscle groups in SMA mice. Neither the position of the muscle in the body, nor the fibre type of the muscle innervated, influenced susceptibility. Morphological properties of vulnerable and disease-resistant motor neurons were then determined from single motor units reconstructed in Thy.1-YFP-H mice. None of the parameters we investigated in healthy young adult mice - including motor unit size, motor unit arbor length, branching patterns, motor endplate size, developmental pruning and numbers of terminal Schwann cells at neuromuscular junctions - correlated with vulnerability. We conclude that morphological characteristics of motor neurons are not a major determinant of disease-susceptibility in SMA, in stark contrast to related forms of motor neuron disease such as ALS. This suggests that subtle molecular differences between motor neurons, or extrinsic factors arising from other cell types, are more likely to determine relative susceptibility in SMA

Comparison of 4- and 5-beam acoustic Doppler current profiler configurations for measurement of turbulent kinetic energy

Acoustic Doppler current profilers (ADCPs) are commonly used to assess mean currents and turbulence at energetic sites. Since 2014, five-beam ADCP configurations have become more common, but conventional analysis of turbulence properties is still based on the four-beam Janus configuration. We use measurements from a single site to investigate improved estimates of turbulent kinetic energy (TKE) that are made possible by the addition of a fifth vertical beam. We conclude that four-beam estimates of TKE are suitable in most cases, and exhibit lower variance than five-beam estimates, but are more prone to contamination by wave activity