Papillomavirus E1 helicase assembly maintains an asymmetric state in the absence of DNA and nucleotide cofactors.

This is a freely-available open access publication. Please cite the published version which is available via the DOI link in this record.Concerted, stochastic and sequential mechanisms of action have been proposed for different hexameric AAA+ molecular motors. Here we report the crystal structure of the E1 helicase from bovine papillomavirus, where asymmetric assembly is for the first time observed in the absence of nucleotide cofactors and DNA. Surprisingly, the ATP-binding sites adopt specific conformations linked to positional changes in the DNA-binding hairpins, which follow a wave-like trajectory, as observed previously in the E1/DNA/ADP complex. The protein's assembly thus maintains such an asymmetric state in the absence of DNA and nucleotide cofactors, allowing consideration of the E1 helicase action as the propagation of a conformational wave around the protein ring. The data imply that the wave's propagation within the AAA+ domains is not necessarily coupled with a strictly sequential hydrolysis of ATP. Since a single ATP hydrolysis event would affect the whole hexamer, such events may simply serve to rectify the direction of the wave's motion.Wellcome TrustYorkshire Cancer ResearchBBSRCEPSRCMR

Exploring the awareness, attitudes, and actions (AAA) of UK adults at high risk of severe illness from COVID-19

Background: People at high risk of severe illness from COVID-19 have experienced greater restrictions during the pandemic, yet there is a paucity of research exploring their lived experience. / Objectives: This study explored the impact of COVID-19 on people identified as at high risk of severe illness by UK Government, and in particular, the impact of the first lockdown on access to healthcare, medications and use of technological platforms. / Methods: 1038 UK adults who identified as at high risk of severe illness from COVID-19 in line with UK Government guidance or self-identified with acute or other chronic health conditions, completed the Awareness, Attitudes and Actions survey which explored the impact of COVID-19 on access to healthcare, management of long-term health condition, mental health, and health behaviours. / Results: Most participants reported feelings of vulnerability, anxiety and isolation, noticed that other people changed their behaviour towards them including a feeling of being stigmatised by people not categorised as high risk. Participants described the largely negative impact that the COVID-19 lockdown had on to health-related behaviours and access to healthcare, which had resulted in large declines in mental health and wellbeing. Participants also indicated disappointment at the UK Governments response and handling of the COVID-19 lockdown. / Implications: This study provides novel evidence of the lived experience of the first COVID-19 lockdown for people identified as at high risk of severe illness. In the context of behavioural health interventions, the ubiquity of digital technologies and their adoption into day-to-day life translates into greater potential reach than traditional interventions, and consequently, greater potential for positive public health impact. Findings should be considered by policymakers and healthcare professionals to support people now and as we transition through the recovery phase with a particular emphasis on supporting mental health and changes to the management of long-term health conditions

The alignment of the second velocity moment tensor in galaxies

We show that provided the principal axes of the second velocity moment tensor of a stellar population are generally unequal and are oriented perpendicular to a set of orthogonal surfaces at each point, then those surfaces must be confocal quadric surfaces and the potential must be separable or Stäckel. This is true under the mild assumption that the even part of the distribution function (DF) is invariant under time reversal vi → −vi of each velocity component. In particular, if the second velocity moment tensor is everywhere exactly aligned in spherical polar coordinates, then the potential must be of separable or Stäckel form (excepting degenerate cases where two or more of the semiaxes of ellipsoid are everywhere the same). The theorem also has restrictive consequences for alignment in cylindrical polar coordinates, which is used in the popular Jeans Anisotropic Models (JAM) of Cappellari. We analyse data on the radial velocities and proper motions of a sample of ∼7300 stars in the stellar halo of the Milky Way. We provide the distributions of the tilt angles or misalignments from both the spherical polar coordinate systems. We show that in this sample the misalignment is always small (usually within 3°) for Galactocentric radii between ∼6 and ∼11 kpc. The velocity anisotropy is very radially biased (β ≈ 0.7), and almost invariant across the volume in our study. Finally, we construct a triaxial stellar halo in a triaxial NFW dark matter halo using a made-to-measure method. Despite the triaxiality of the potential, the velocity ellipsoid of the stellar halo is nearly spherically aligned within ∼6° for large regions of space, particularly outside the scale radius of the stellar halo. We conclude that the second velocity moment ellipsoid can be close to spherically aligned for a much wider class of potentials than the strong constraints that arise from exact alignment might suggest

Using a simple expert system to assist a powered wheelchair user

A simple expert system is described that helps wheelchair users to drive their wheelchairs. The expert system takes data in from sensors and a joystick, identifies obstacles and then recommends a safe route. Wheelchair users were timed while driving around a variety of routes and using a joystick controlling their wheelchair via the simple expert system. Ultrasonic sensors are used to detect the obstacles. The simple expert system performed better than other recently published systems. In more difficult situations, wheelchair drivers did better when there was help from a sensor system. Wheelchair users completed routes with the sensors and expert system and results are compared with the same users driving without any assistance. The new systems show a significant improvement

NMR studies of p7 protein from hepatitis C virus

The p7 protein of hepatitis C virus (HCV) plays an important role in the viral lifecycle. Like other members of the viroporin family of small membrane proteins, the amino acid sequence of p7 is largely conserved over the entire range of genotypes, and it forms ion channels that can be blocked by a number of established channel-blocking compounds. Its characteristics as a membrane protein make it difficult to study by most structural techniques, since it requires the presence of lipids to fold and function properly. Purified p7 can be incorporated into phospholipid bilayers and micelles. Initial solid-state nuclear magnetic resonance (NMR) studies of p7 in 14-O-PC/6-O-PC bicelles indicate that the protein contains helical segments that are tilted approximately 10° and 25° relative to the bilayer normal. A truncated construct corresponding to the second transmembrane domain of p7 is shown to have properties similar to those of the full-length protein, and was used to determine that the helix segment tilted at 10° is in the C-terminal portion of the protein. The addition of the channel blocker amantadine to the full-length protein resulted in selective chemical shift changes, demonstrating that NMR has a potential role in the development of drugs targeted to p7

Methanomethylovorans are the dominant dimethylsulfide-degrading methanogens in gravel and sandy river sediment microcosms

Abstract Background Rivers and streams are important components of the global carbon cycle and methane budget. However, our understanding of the microbial diversity and the metabolic pathways underpinning methylotrophic methane production in river sediments is limited. Dimethylsulfide is an important methylated compound, found in freshwater sediments. Yet, the magnitude of DMS-dependent methanogenesis nor the methanogens carrying out this process in river sediments have been explored before. This study addressed this knowledge gap in DMS-dependent methanogenesis in gravel and sandy river sediments. Results Significant methane production via DMS degradation was found in all sediment  microcosms. Sandy, less permeable river sediments had higher methane yields (83 and 92%) than gravel, permeable sediments (40 and 48%). There was no significant difference between the methanogen diversity in DMS-amended gravel and sandy sediment microcosms, which Methanomethylovorans dominated. Metagenomics data analysis also showed the dominance of Methanomethylovorans and Methanosarcina. DMS-specific methyltransferase genes (mts) were found in very low relative abundances whilst the methanol-, trimethylamine- and dimethylamine-specific methyltransferase genes (mtaA, mttB and mtbB) had the highest relative abundances, suggesting their involvement in DMS-dependent methanogenesis. Conclusions This is the first study demonstrating a significant potential for DMS-dependent methanogenesis in river sediments with contrasting geologies. Methanomethylovorans were the dominant methylotrophic methanogen in all river sediment microcosms. Methyltransferases specific to methylotrophic substrates other than DMS are likely key enzymes in DMS-dependent methanogenesis, highlighting their versatility and importance in the methane cycle in freshwater sediments, which would warrant further study. </jats:sec

Regional Heterogeneity in Murine Lung Fibroblasts from Normal Mice or Mice Exposed Once to Cigarette Smoke

Chronic obstructive lung disease (COPD) is characterized by matrix deposition in the small airways but matrix loss from the parenchyma, phenomena which must depend on the ability of local fibroblasts to produce matrix after smoke exposure. To investigate this idea, we exposed C57Bl/6 mice once to cigarette smoke or to air (control) and prepared primary cultures of lung fibroblasts by microdissecting large airways (trachea, LAF), medium size airways (major bronchi, MAF) and parenchyma (PF). Control PF showed the lowest rate of wound closure and wound closure was depressed in all lines by a single in vivo smoke exposure. Gene expression of matrix proteins differed considerably among the sites; decorin, which may sequester TGFβ, was markedly higher in PF. PF showed higher intrinsic ratios of pSmad2/Smad2. Smoke caused much greater increases in secreted and matrix deposited collagens 1 and 3 in PF than in LAF or MAF. Expression of Thy-1, a gene that suppresses myofibroblast differentiation, was increased by smoke in PF. We conclude that there is considerable regional heterogeneity in murine lung fibroblasts in terms of matrix production, either basally or after in vivo smoke exposure; that PF have lower ability to repair wounds and higher intrinsic TGFβ signaling; and that a single exposure to smoke produces lasting changes in the pattern of matrix production and wound repair, changes that may be mediated in part by smoke-induced release of TGFβ. However, PF still retain the ability to repair by producing new matrix after a single in vivo smoke exposure

Violent and non-violent crimes against sex workers : the influence of the sex market on reporting practices in the United Kingdom

Previous research has shown that sex workers experience extremely high rates of victimization but are often reluctant to report their experiences to the police. This paper explores how the markets in which sex workers operate in the United Kingdom impact upon the violent and non-violent crimes they report to a national support organization and their willingness to report victimization to the police. We use a secondary quantitative data analysis of 2,056 crime reports submitted to the UK National Ugly Mugs (NUM) scheme between 2012 and 2016. The findings indicate that although violence is the most common crime type reported to NUM, sex workers operating in different markets report varying relative proportions of different types of victimization. We also argue that there is some variation in the level of willingness to share reports with the police across the different sex markets, even when the type crime, presence of violence, and other variables are taken into account. Our finding that street sex workers are most likely to report victimization directly to the police challenges previously held assumptions that criminalization is the key factor preventing sex workers from engaging with the police. Key words: sex work; violence; policing; reported victimizatio

Spin Caloritronics

This is a brief overview of the state of the art of spin caloritronics, the science and technology of controlling heat currents by the electron spin degree of freedom (and vice versa).Comment: To be published in "Spin Current", edited by S. Maekawa, E. Saitoh, S. Valenzuela and Y. Kimura, Oxford University Pres