On the use of ultracentrifugal devices for routine sample preparation in biomolecular magic-angle-spinning NMR

A number of recent advances in the field of magic-angle-spinning (MAS) solid-state NMR have enabled its application to a range of biological systems of ever increasing complexity. To retain biological relevance, these samples are increasingly studied in a hydrated state. At the same time, experimental feasibility requires the sample preparation process to attain a high sample concentration within the final MAS rotor. We discuss these considerations, and how they have led to a number of different approaches to MAS NMR sample preparation. We describe our experience of how custom-made (or commercially available) ultracentrifugal devices can facilitate a simple, fast and reliable sample preparation process. A number of groups have since adopted such tools, in some cases to prepare samples for sedimentation-style MAS NMR experiments. Here we argue for a more widespread adoption of their use for routine MAS NMR sample preparation

Mitochondrial phylogeography and demographic history of the Vicuña: implications for conservation

The vicuña (Vicugna vicugna; Miller, 1924) is a conservation success story, having recovered from near extinction in the 1960s to current population levels estimated at 275 000. However, lack of information about its demographic history and genetic diversity has limited both our understanding of its recovery and the development of science-based conservation measures. To examine the evolution and recent demographic history of the vicuña across its current range and to assess its genetic variation and population structure, we sequenced mitochondrial DNA from the control region (CR) for 261 individuals from 29 populations across Peru, Chile and Argentina. Our results suggest that populations currently designated as Vicugna vicugna vicugna and Vicugna vicugna mensalis comprise separate mitochondrial lineages. The current population distribution appears to be the result of a recent demographic expansion associated with the last major glacial event of the Pleistocene in the northern (18 to 22°S) dry Andes 14–12 000 years ago and the establishment of an extremely arid belt known as the 'Dry Diagonal' to 29°S. Within the Dry Diagonal, small populations of V. v. vicugna appear to have survived showing the genetic signature of demographic isolation, whereas to the north V. v. mensalis populations underwent a rapid demographic expansion before recent anthropogenic impacts

Spectropolarimetry of Supernovae

Overwhelming evidence has accumulated in recent years that supernova explosions are intrinsically 3-dimensional phenomena with significant departures from spherical symmetry. We review the evidence derived from spectropolarimetry that has established several key results: virtually all supernovae are significantly aspherical near maximum light; core-collapse supernovae behave differently than thermonuclear (Type Ia) supernovae; the asphericity of core-collapse supernovae is stronger in the inner layers showing that the explosion process itself is strongly aspherical; core-collapse supernovae tend to establish a preferred direction of asymmetry; the asphericity is stronger in the outer layers of thermonuclear supernovae providing constraints on the burning process. We emphasize the utility of the Q/U plane as a diagnostic tool and revisit SN 1987A and SN 1993J in a contemporary context. An axially-symmetric geometry can explain many basic features of core-collapse supernovae, but significant departures from axial symmetry are needed to explain most events. We introduce a spectropolarimetry type to classify the range of behavior observed in polarized supernovae. Understanding asymmetries in supernovae is important for phenomena as diverse as the origins of gamma-ray bursts and the cosmological applications of Type Ia supernovae in studies of the dark energy content of the universe.Comment: Draft of Annual Review article prior to final copy editing; 85 pages, 13 figures, 1 tabl

Is remaining indoors an effective way of reducing exposure to fine particulate matter during biomass burning events?

Bushfires, prescribed burns, and residential wood burning are significant sources of fine particles (aerodynamic diameter 2.5) affecting the health and well-being of many communities. Despite the lack of evidence, a common public health recommendation is to remain indoors, assuming that the home provides a protective barrier against ambient PM2.5. The study aimed to assess to what extent houses provide protection against peak concentrations of outdoor PM2.5 and whether remaining indoors is an effective way of reducing exposure to PM2.5. The effectiveness of this strategy was evaluated by conducting simultaneous week-long indoor and outdoor measurements of PM2.5 at 21 residences in regional areas of Victoria, Australia. During smoke plume events, remaining indoors protected residents from peak outdoor PM2.5 concentrations, but the level of protection was highly variable, ranging from 12% to 76%. Housing stock (e.g., age of the house) and ventilation (e.g., having windows/doors open or closed) played a significant role in the infiltration of outdoor PM2.5 indoors. The results also showed that leaving windows and doors closed once the smoke plume abates trapped PM2.5 indoors and increased indoor exposure to PM2.5. Furthermore, for approximately 50% of households, indoor sources such as cooking activities, smoking, and burning candles or incense contributed significantly to indoor PM2.5. Implications: Smoke from biomass burning sources can significantly impact on communities. Remaining indoors with windows and doors closed is a common recommendation by health authorities to minimize exposures to peak concentrations of fine particles during smoke plume events. Findings from this study have shown that the protection from fine particles in biomass burning smoke is highly variable among houses, with information on housing age and ventilation status providing an approximate assessment on the protection of a house. Leaving windows closed once a smoke plume abates traps particles indoors and increases exposures

RINGO3 polarimetry of the Type I superluminous SN 2017egm

The origin of the luminosity of superluminous supernovae (SLSNe) is an unresolved mystery, and a number of very different physical scenarios (including energy injection from magnetars, collision with a dense circumstellar medium, and pair instability-induced explosions) have been invoked. The application of polarimetry to normal SNe has been shown to probe the three-dimensional structure of exploding stars, providing clues to the nature of the explosion mechanism. We report imaging linear polarimetry observations of the Type I SLSN 2017egm, in the galaxy NGC 3191, conducted with the Liverpool Telescope and the RINGO3 instrument. Observations were acquired at four epochs, spanning 4–19 d after light curve maximum, however, polarization was not detected at a level of >3σ. At +7 and + 15 d, and in the average over all epochs, we find a possible polarization signal, detected at a significance of ≈2σ in the ‘blue’ channel. This signal is seen, primarily, in the Stokes q parameter, with a corresponding polarization angle consistent with the orientation of the spiral arm in proximity to the position of SN 2017egm. We interpret this as indicating that any polarization, if present, originates from dust in the host galaxy rather than being intrinsic to the SN itself. Despite its apparent peculiarities, compared to other Type I SLSNe, the polarization characteristics of SN 2017egm are consistent with the previously reported low polarization of other SLSNe of this variety

Defining functional interactions during biogenesis of epithelial junctions

In spite of extensive recent progress, a comprehensive understanding of how actin cytoskeleton remodelling supports stable junctions remains to be established. Here we design a platform that integrates actin functions with optimized phenotypic clustering and identify new cytoskeletal proteins, their functional hierarchy and pathways that modulate E-cadherin adhesion. Depletion of EEF1A, an actin bundling protein, increases E-cadherin levels at junctions without a corresponding reinforcement of cell-cell contacts. This unexpected result reflects a more dynamic and mobile junctional actin in EEF1A-depleted cells. A partner for EEF1A in cadherin contact maintenance is the formin DIAPH2, which interacts with EEF1A. In contrast, depletion of either the endocytic regulator TRIP10 or the Rho GTPase activator VAV2 reduces E-cadherin levels at junctions. TRIP10 binds to and requires VAV2 function for its junctional localization. Overall, we present new conceptual insights on junction stabilization, which integrate known and novel pathways with impact for epithelial morphogenesis, homeostasis and diseases

Celecoxib exerts protective effects in the vascular endothelium via COX-2-independent activation of AMPK-CREB-Nrf2 signalling

Although concern remains about the athero-thrombotic risk posed by cyclo-oxygenase (COX)-2-selective inhibitors, recent data implicates rofecoxib, while celecoxib appears equivalent to NSAIDs naproxen and ibuprofen. We investigated the hypothesis that celecoxib activates AMP kinase (AMPK) signalling to enhance vascular endothelial protection. In human arterial and venous endothelial cells (EC), and in contrast to ibuprofen and naproxen, celecoxib induced the protective protein heme oxygenase-1 (HO-1). Celecoxib derivative 2,5-dimethyl-celecoxib (DMC) which lacks COX-2 inhibition also upregulated HO-1, implicating a COX-2-independent mechanism. Celecoxib activated AMPKα(Thr172) and CREB-1(Ser133) phosphorylation leading to Nrf2 nuclear translocation. Importantly, these responses were not reproduced by ibuprofen or naproxen, while AMPKα silencing abrogated celecoxib-mediated CREB and Nrf2 activation. Moreover, celecoxib induced H-ferritin via the same pathway, and increased HO-1 and H-ferritin in the aortic endothelium of mice fed celecoxib (1000 ppm) or control chow. Functionally, celecoxib inhibited TNF-α-induced NF-κB p65(Ser536) phosphorylation by activating AMPK. This attenuated VCAM-1 upregulation via induction of HO-1, a response reproduced by DMC but not ibuprofen or naproxen. Similarly, celecoxib prevented IL-1β-mediated induction of IL-6. Celecoxib enhances vascular protection via AMPK-CREB-Nrf2 signalling, a mechanism which may mitigate cardiovascular risk in patients prescribed celecoxib. Understanding NSAID heterogeneity and COX-2-independent signalling will ultimately lead to safer anti-inflammatory drugs

Individual variation in levels of haptoglobin-related protein in children from Gabon

Background: Haptoglobin related protein (Hpr) is a key component of trypanosome lytic factors (TLF), a subset of highdensity lipoproteins (HDL) that form the first line of human defence against African trypanosomes. Hpr, like haptoglobin (Hp) can bind to hemoglobin (Hb) and it is the Hpr-Hb complexes which bind to these parasites allowing uptake of TLF. This unique form of innate immunity is primate-specific. To date, there have been no population studies of plasma levels of Hpr, particularly in relation to hemolysis and a high prevalence of ahaptoglobinemia as found in malaria endemic areas. Methods and Principal Findings: We developed a specific enzyme-linked immunosorbent assay to measure levels of plasma Hpr in Gabonese children sampled during a period of seasonal malaria transmission when acute phase responses (APR), malaria infection and associated hemolysis were prevalent. Median Hpr concentration was 0.28 mg/ml (range 0.03-1.1). This was 5-fold higher than that found in Caucasian children (0.049 mg/ml, range 0.002-0.26) with no evidence of an APR. A general linear model was used to investigate associations between Hpr levels, host polymorphisms, parasitological factors and the acute phase proteins, Hp, C-reactive protein (CRP) and albumin. Levels of Hpr were associated with Hp genotype, decreased with age and were higher in females. Hpr concentration was strongly correlated with that of Hp, but not CRP

The longitudinal NIHR ARC North West Coast Household Health Survey: exploring health inequalities in disadvantaged communities.

BACKGROUND: The Household Health Survey (HHS) was developed to understand the socioeconomic determinants of mental and physical health, and health inequalities in health and social care. This paper aims to provide a detailed rationale of the development and implementation of the survey and explore socio-economic variations in physical and mental health and health care. METHODS: This comprehensive longitudinal public health survey was designed and piloted in a disadvantaged area of England, comprising questions on housing, physical health, mental health, lifestyle, social issues, environment, work, and finances. After piloting, the HHS was implemented across 28 neighbourhoods - 10 disadvantaged neighbourhoods for learning (NfLs), 10 disadvantaged comparator sites, and eight relatively advantaged areas, in 2015 and 2018. Participants were recruited via random sampling of households in pre-selected neighbourhoods based on their areas of deprivation. RESULTS: 7731 residents participated in Wave 1 (N = 4319) and 2 (n = 3412) of the survey, with 871 residents having participated in both. Mental health, physical health, employment, and housing quality were poorer in disadvantaged neighbourhoods than in relatively advantaged areas. CONCLUSIONS: This survey provides important insights into socio-economic variations in physical and mental health, with findings having implications for improved care provision to enable residents from any geographical or socio-economic background to access suitable care

Hubble Space Telescope and ground-based observations of SN 1993J and SN 1998S: CNO processing in the progenitors

Ground-based and Hubble Space Telescope observations are presented for SN 1993J and SN 1998S. SN 1998S shows strong, relatively narrow circumstellar emission lines of N III-V and C III-IV, as well as broad lines from the ejecta. Both the broad ultraviolet and optical lines in SN 1998S indicate an expansion velocity of ∼7000 km s -1. The broad emission components of Lyα and Mg II are strongly asymmetrical after day 72 past the explosion and differ in shape from Hα. Different models based on dust extinction from dust in the ejecta or shock region, in combination with Hα from a circumstellar torus, are discussed. It is concluded, however, that the double-peaked line profiles are more likely to arise as a result of optical depth effects in the narrow, cool, dense shell behind the reverse shock than in a torus-like region. The ultraviolet lines of SN 1993J are broad, with a boxlike shape, coming from the ejecta and a cool, dense shell. The shapes of the lines are well fitted by a shell with inner velocity ∼7000 km s -1 and outer velocity ∼10,000 km s -1. For both SN 1993J and SN 1998S a strong nitrogen enrichment is found, with N/C ≈ 12.4 in SN 1993J and N/C ≈ 6.0 in SN 1998S. From a compilation of all supernovae with determined CNO ratios, we discuss the implications of these observations for the structure of the progenitors of Type II supernovae. © 2005. The American Astronomical Society. All rights reserved,.published_or_final_versio