The cellular prion protein traps Alzheimer's A beta in an oligomeric form and disassembles amyloid fibers

This work was supported by a Wellcome Trust project grant (093241/Z/10/Z) and UK Biotechnology and Biological Sciences Research Council Quota studentships. The authors thank Harold Toms (Queen Mary, University of London) and the UK National Institute for Medical Research for NMR support, and Graham McPhail for assistance with TEM

p62 overexpression induces TDP-43 cytoplasmic mislocalisation, aggregation and cleavage and neuronal death

Amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD) that exist on a spectrum of neurodegenerative disease. A hallmark of pathology is cytoplasmic TDP-43 aggregates within neurons, observed in 97% of ALS cases and ~ 50% of FTLD cases. This mislocalisation from the nucleus into the cytoplasm and TDP-43 cleavage are associated with pathology, however, the drivers of these changes are unknown. p62 is invariably also present within these aggregates. We show that p62 overexpression causes TDP-43 mislocalisation into cytoplasmic aggregates, and aberrant TDP-43 cleavage that was dependent on both the PB1 and ubiquitin-associated (UBA) domains of p62. We further show that p62 overexpression induces neuron death. We found that stressors (proteasome inhibition and arsenic) increased p62 expression and that this shifted the nuclear:cytoplasmic TDP-43 ratio. Overall, our study suggests that environmental factors that increase p62 may thereby contribute to TDP-43 pathology in ALS and FTLD

Alpha-2-Macroglobulin Is Acutely Sensitive to Freezing and Lyophilization: Implications for Structural and Functional Studies.

Alpha-2-macroglobulin is an abundant secreted protein that is of particular interest because of its diverse ligand binding profile and multifunctional nature, which includes roles as a protease inhibitor and as a molecular chaperone. The activities of alpha-2-macroglobulin are typically dependent on whether its conformation is native or transformed (i.e. adopts a more compact conformation after interactions with proteases or small nucleophiles), and are also influenced by dissociation of the native alpha-2-macroglobulin tetramer into stable dimers. Alpha-2-macroglobulin is predominately present as the native tetramer in vivo; once purified from human blood plasma, however, alpha-2-macroglobulin can undergo a number of conformational changes during storage, including transformation, aggregation or dissociation. We demonstrate that, particularly in the presence of sodium chloride or amine containing compounds, freezing and/or lyophilization of alpha-2-macroglobulin induces conformational changes with functional consequences. These conformational changes in alpha-2-macroglobulin are not always detected by standard native polyacrylamide gel electrophoresis, but can be measured using bisANS fluorescence assays. Increased surface hydrophobicity of alpha-2-macroglobulin, as assessed by bisANS fluorescence measurements, is accompanied by (i) reduced trypsin binding activity, (ii) increased chaperone activity, and (iii) increased binding to the surfaces of SH-SY5Y neurons, in part, via lipoprotein receptors. We show that sucrose (but not glycine) effectively protects native alpha-2-macroglobulin from denaturation during freezing and/or lyophilization, thereby providing a reproducible method for the handling and long-term storage of this protein.Early Career Fellowship from the National Health and Medical Research Council GNT1012521(A.R.W.); Wellcome Trust Programme Grant (J.R.K., C.M.D.) 094425/Z/10/Z; Samsung GRO Grant (M.R.W.)This is the final version of the article. It first appeared from PLoS via http://dx.doi.org/10.1371/journal.pone.013003

Evaluating protein cross-linking as a therapeutic strategy to stabilize SOD1 variants in a mouse model of familial ALS

Mutations in the gene encoding Cu-Zn superoxide dismutase 1 (SOD1) cause a subset of familial amyotrophic lateral sclerosis (fALS) cases. A shared effect of these mutations is that SOD1, which is normally a stable dimer, dissociates into toxic monomers that seed toxic aggregates. Considerable research effort has been devoted to developing compounds that stabilize the dimer of fALS SOD1 variants, but unfortunately, this has not yet resulted in a treatment. We hypothesized that cyclic thiosulfinate cross-linkers, which selectively target a rare, 2 cysteine-containing motif, can stabilize fALS-causing SOD1 variants in vivo. We created a library of chemically diverse cyclic thiosulfinates and determined structure-cross-linking-activity relationships. A pre-lead compound, “S-XL6,” was selected based upon its cross-linking rate and drug-like properties. Co-crystallographic structure clearly establishes the binding of S-XL6 at Cys 111 bridging the monomers and stabilizing the SOD1 dimer. Biophysical studies reveal that the degree of stabilization afforded by S-XL6 (up to 24°C) is unprecedented for fALS, and to our knowledge, for any protein target of any kinetic stabilizer. Gene silencing and protein degrading therapeutic approaches require careful dose titration to balance the benefit of diminished fALS SOD1 expression with the toxic loss-of-enzymatic function. We show that S-XL6 does not share this liability because it rescues the activity of fALS SOD1 variants. No pharmacological agent has been proven to bind to SOD1 in vivo. Here, using a fALS mouse model, we demonstrate oral bioavailability; rapid engagement of SOD1G93A by S-XL6 that increases SOD1G93A’s in vivo half-life; and that S-XL6 crosses the blood–brain barrier. S-XL6 demonstrated a degree of selectivity by avoiding off-target binding to plasma proteins. Taken together, our results indicate that cyclic thiosulfinate-mediated SOD1 stabilization should receive further attention as a potential therapeutic approach for fALS

Defining Terms Used for Animals Working in Support Roles for People with Support Needs

This is the final version. Available on open access from MDPI via the DOI in this recordData Availability Statement: De-identified qualitative data can be made available by contacting the lead author.The nomenclature used to describe animals working in roles supporting people can be confusing. The same term may be used to describe different roles, or two terms may mean the same thing. This confusion is evident among researchers, practitioners, and end users. Because certain animal roles are provided with legal protections and/or government-funding support in some jurisdictions, it is necessary to clearly define the existing terms to avoid confusion. The aim of this paper is to provide operationalized definitions for nine terms, which would be useful in many world regions: "assistance animal", "companion animal", "educational/school support animal", "emotional support animal", "facility animal", "service animal", "skilled companion animal", "therapy animal", and "visiting/visitation animal". At the International Society for Anthrozoology (ISAZ) conferences in 2018 and 2020, over 100 delegates participated in workshops to define these terms, many of whom co-authored this paper. Through an iterative process, we have defined the nine terms and explained how they differ from each other. We recommend phasing out two terms (i.e., "skilled companion animal" and "service animal") due to overlap with other terms that could potentially exacerbate confusion. The implications for several regions of the world are discussed

The spatial and temporal distribution of moisture within Sitka spruce standing trees and roundwood logs

Within the UK, levels of timber processing are set to increase as the plantation grown forest resource reaches financial maturity. Made up predominately of exotic coniferous species, this resource has been the focus of many recent studies as wood users look to improve their fundamental knowledge of the primary characteristics that will enable them to make more appropriate decisions and capitalise on future opportunities. As a further contribution to this knowledge base, it was the aim of this thesis to determine moisture content variation within Sitka spruce (Picea sitchensis (Bong.) Carr) standing trees and roundwood logs. Within standing trees, moisture content was found to vary significantly in both the longitudinal and radial directions (P<0.001). Longitudinally, the base and top of the trees contained higher quantities of moisture than did the lower and mid sections. Radially, the moisture content at the centre of each disk was very low and this extended outwards before rapidly increasing mid-radius and then reaching a plateau near to the cambium. A sigmoid regression model successfully explained two-thirds of the radial moisture content variation as well as explaining the longitudinal moisture content variation. The radial variation with tree height was described by a shortening of the low moisture content extending outwards from the pith and a softening of the steepness of the increase with the duration extending as tree height increased. Seasonal variation within standing spruce trees was assessed within a plantation forest in northern England. Removing samples from 81 trees throughout a period covering 42 calendar weeks of 2012 revealed no significant effect of season on the moisture content although a link was identified with short-term variation in temperature. However, further analysis on the radial sections confirmed that when the moisture content data were converted to a percentage of total lumen saturation, a significant effect of seasonal variation was observed within the outer sapwood region (P<0.001). Whilst the standing trees as a whole did not demonstrate seasonal variation in moisture content, variation due to seasonal weather did have an effect on the drying of roundwood within the forest. Analysis of c. 25,000 samples, collected over a 6-year period, confirmed that season (dormant or growing) had a significant effect on sample moisture content (P<0.001). Temperature was recorded as having the greatest effect on moisture variation within roundwood whilst the effect of rainfall was not significant, due principally to the unpredictable nature of this variable throughout the year. To identify the radial and longitudinal moisture content change of roundwood during drying, an experiment was carried out to accelerate the drying process on short logs using an industrial oven at a constant temperature (40°C). Total drying time varied between 7 and 28 days prior to post drying analysis of the logs to determine the longitudinal and radial moisture profiles. All logs lost a constant quantity of moisture each day with the logs dried for 28 days losing more than 50% of their starting weight. Longitudinally, moisture was lost from all points along the length of each log with the log ends remaining driest, although these dry areas only accounted for 40% of the total log length. Radially, moisture declined across the log with the greatest amount removed from the outer radius. In order to provide some relevance of the impact moisture content variation has on the forest industry value chain, data collected from a working timber harvesting operation confirmed that variation existed between volume to weight conversions of different products. Within this even-aged Sitka spruce clearfell, volume to weight conversion factors ranged from 1.02 to 1.19. Differences of this magnitude can not only lead to a lack of understanding of between tree variation within a single site, but can also result in an impact on the optimum value recovery expected by each of the parties engaged within the value chain. The results presented within this thesis will help guide timber growers, agents, managers and purchasers in their future decision making when considering the most suitable markets for standing trees or roundwood logs. Improving knowledge of spatial and temporal moisture content variation will potentially result in maximising the forestry value chain and enable the forestry sector in the UK to make more from wood