A new automated technique for the reconstitution of hydrophobic proteins into planar bilayer membranes. Studies of human recombinant uncoupling protein 1

AbstractElectrophysiological characterisation of the vast number of annotated channel and transport proteins in the postgenomic era would be greatly facilitated by the introduction of rapid and robust methods for the functional incorporation of membrane proteins into defined lipid bilayers. Here, we describe an automated technique for reconstitution of membrane proteins into lipid bilayer membranes, which substantially reduces both the reconstitution time and the amount of protein required for the membrane formation. The method allows the investigation of single protein channels as well as insertion of multiple copies (∼107) into a single bilayer. Despite a comparatively large membrane area (up to 300 μm diameter), the high stability of the membrane permits the application of transmembrane voltages up to 300 mV. This feature is especially important for studies of inner membrane mitochondrial proteins, since they act at potentials up to ∼200 mV under physiological conditions. It is a combination of these advantages that enables the detailed investigation of the minuscule single protein conductances typical for proton transporters. We have applied the new technique for the reconstitution and electrophysiological characterisation of human recombinant uncoupling protein 1, hUCP1, that has been overexpressed in E. coli and purified from inclusion bodies. We demonstrate that hUCP1 activity in the presence of fatty acids is comparable to the activity of UCP1 isolated from brown adipose tissue

Superconducting and normal-state properties of the noncentrosymmetric superconductor Re3Ta

The noncentrosymmetric superconductor, Re3Ta, has been characterized in detail with a combination of magnetization, heat capacity, and electrical resistivity measurements, as well as a microscopic investigation of the internal magnetic fields using muon spin spectroscopy (μSR). In low applied fields, we observe 100% flux expulsion at a temperature of Tc = 4.68 K, which is concomitant with a sudden decrease of the electrical resistivity to zero and a sharp discontinuity in the heat capacity, confirming bulk superconductivity in this material. We find that Re3Ta is a poor metal, with superconductivity occurring in the dirty limit, and in which the disorder in the structure dominates the physical properties. Zero-field μSR shows that the superconducting state preserves time-reversal symmetry, and transverse-field measurements of the superfluid density are well described by an isotropic s-wave model. A careful analysis of the internal field distribution reveals a high level of disorder in the vortex lattice. Furthermore, we have combined the experimental data and calculated the effective mass, carrier density, and electronic mean-free path in this material, and ultimately show that Re3Ta lies close to the unconventional region of the Uemura plot

Gene expression analysis of TIL rich HPV-driven head and neck tumors reveals a distinct B-cell signature when compared to HPV independent tumors

Human papilloma virus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) has a better prognosis than it's HPV negative (HPV(-)) counterpart. This may be due to the higher numbers of tumor-infiltrating lymphocytes (TILs) in HPV positive (HPV(+)) tumors. RNA-Sequencing (RNA-Seq) was used to evaluate whether the differences in clinical behaviour simply reflect a numerical difference in TILs or whether there is a fundamental behavioural difference between TILs in these two settings. Thirty-nine HNSCC tumors were scored for TIL density by immunohistochemistry. After the removal of 16 TILlow tumors, RNA-Seq analysis was performed on 23 TILhigh/med tumors (HPV(+) n=10 and HPV(-) n=13). Using EdgeR, differentially expressed genes (DEG) were identified. Immune subset analysis was performed using Functional Analysis of Individual RNA-Seq/ Microarray Expression (FAIME) and immune gene RNA transcript count analysis. In total, 1,634 DEGs were identified, with a dominant immune signature observed in HPV(+) tumors. After normalizing the expression profiles to account for differences in B- and T-cell number, 437 significantly DEGs remained. A B-cell associated signature distinguished HPV(+) from HPV(-) tumors, and included the DEGs CD200, GGA2, ADAM28, STAG3, SPIB, VCAM1, BCL2 and ICOSLG; the immune signal relative to T-cells was qualitatively similar between TILs of both tumor cohorts. Our findings were validated and confirmed in two independent cohorts using TCGA data and tumor-infiltrating B-cells from additional HPV(+) HNSCC patients. A B-cell associated signal segregated tumors relative to HPV status. Our data suggests that the role of B-cells in the adaptive immune response to HPV(+) HNSCC requires re-assessment

A new framework to enable equitable outcomes: resilience and nexus approaches combined

Managing integrated social-ecological systems to reduce risks to human and environmental well-being remains challenging in light of the rate and extent of undesirable changes that are occurring. Developing frameworks that are sufficiently integrative to guide research to deliver the necessary insights into all key system aspects is an important outstanding task. Among existing approaches, resilience and nexus framings both allow focus on unpacking relationships across scales and levels in a system and emphasize the involvement of different groups in decision making to different extents. They also suffer weaknesses and neither approach puts social justice considerations explicitly at its core. This has important implications for understanding who wins and loses out from different decisions and how social and ecological risks and trade-offs are shared and distributed, temporally and spatially. This paper conceptually integrates resilience and nexus approaches, developing a combined framework and indicating how it could effectively be operationalized in cases from mountain and mangrove social-ecological systems. In doing so, it advances understanding of complex social-ecological systems framings for risk-based decision making beyond that which could be achieved through use of either resilience or nexus approaches alone. Important next steps in testing the framework involve empirical and field operationalization, requiring interdisciplinary, mixed method approache

ACCESS II: A Complete Census of Star Formation in the Shapley Supercluster - UV and IR Luminosity Functions

We present panoramic Spitzer/MIPS mid- and far-infrared and GALEX ultraviolet imaging of the the most massive and dynamically active system in the local Universe, the Shapley supercluster at z=0.048, covering the 5 clusters which make up the supercluster core. We combine these data with existing spectroscopic data from 814 confirmed supercluster members to produce the first study of a local rich cluster including both ultraviolet and infrared luminosity functions (LFs). This joint analysis allows us to produce a complete census of star-formation (both obscured and unobscured), extending down to SFRs~0.02-0.05Msun/yr, and quantify the level of obscuration of star formation among cluster galaxies, providing a local benchmark for comparison to ongoing and future studies of cluster galaxies at higher redshifts with Spitzer and Herschel. The GALEX NUV and FUV LFs obtained have steeper faint-end slopes than the local field population, due largely to the contribution of massive, quiescent galaxies at M_FUV>-16. The 24um and 70um galaxy LFs for the Shapley supercluster instead have shapes fully consistent with those obtained for the Coma cluster and for the local field galaxy population. This apparent lack of environmental dependence for the shape of the FIR luminosity function suggests that the bulk of the star-forming galaxies that make up the observed cluster infrared LF have been recently accreted from the field and have yet to have their star formation activity significantly affected by the cluster environment. We estimate a global SFR of 327 Msun/yr over the whole supercluster core, of which just ~20% is visible directly in the UV continuum and ~80% is reprocessed by dust and emitted in the infrared. The level of obscuration (L_IR/L_FUV) in star-forming galaxies is seen to increase linearly with L_K over two orders of magnitude in stellar mass.Comment: 19 pages, 17 figures. Accepted for publication in MNRA

The <i>Herschel</i> view of the massive star-forming region NGC 6334

Aims: Fundamental to any theory of high-mass star formation are gravity and turbulence. Their relative importance, which probably changes during cloud evolution, is not known. By investigating the spatial and density structure of the high-mass star-forming complex NGC 6334 we aim to disentangle the contributions of turbulence and gravity. Methods: We used Herschel PACS and SPIRE imaging observations from the HOBYS key programme at wavelengths of 160, 250, 350, and 500 μm to construct dust temperature and column density maps. Using probability distribution functions (PDFs) of the column density determined for the whole complex and for four distinct sub-regions (distinguished on the basis of differences in the column density, temperature, and radiation field), we characterize the density structure of the complex. We investigate the spatial structure using the Δ-variance, which probes the relative amount of structure on different size scales and traces possible energy injection mechanisms into the molecular cloud. Results: The Δ-variance analysis suggests that the significant scales of a few parsec that were found are caused by energy injection due to expanding HII regions, which are numerous, and by the lengths of filaments seen everywhere in the complex. The column density PDFs have a lognormal shape at low densities and a clearly defined power law at high densities for all sub-regions whose slope is linked to the exponent α of an equivalent spherical density distribution. In particular with α = 2.37, the central sub-region is largly dominated by gravity, caused by individual collapsing dense cores and global collapse of a larger region. The collapse is faster than free-fall (which would lead only to α = 2) and thus requires a more dynamic scenario (external compression, flows). The column density PDFs suggest that the different sub-regions are at different evolutionary stages, especially the central sub-region, which seems to be in a more evolved stage

MALT-45: a 7 mm survey of the southern Galaxy - I. Techniques and spectral line data

We present the first results from the MALT-45 (Millimetre Astronomer's Legacy Team-45 GHz) Galactic Plane survey. We have observed 5 square degrees (l = 330°–335°, b = ±0 ∘ . 5) for spectral lines in the 7 mm band (42–44 and 48–49 GHz), including CS (1–0), class I CH3OH masers in the 7(0,7)–6(1,6) A+ transition and SiO (1–0) v = 0, 1, 2, 3. MALT-45 is the first unbiased, large-scale, sensitive spectral line survey in this frequency range. In this paper, we present data from the survey as well as a few intriguing results; rigorous analyses of these science cases are reserved for future publications. Across the survey region, we detected 77 class I CH3OH masers, of which 58 are new detections, along with many sites of thermal and maser SiO emission and thermal CS. We found that 35 class I CH3OH masers were associated with the published locations of class II CH3OH, H2O and OH masers but 42 have no known masers within 60 arcsec. We compared the MALT-45 CS with NH3 (1,1) to reveal regions of CS depletion and high opacity, as well as evolved star-forming regions with a high ratio of CS to NH3. All SiO masers are new detections, and appear to be associated with evolved stars from the Spitzer Galactic Legacy Infrared Mid-Plane Survey Extraordinaire (GLIMPSE). Generally, within SiO regions of multiple vibrational modes, the intensity decreases as v = 1, 2, 3, but there are a few exceptions where v = 2 is stronger than v = 1

International meta-analysis of PTSD genome-wide association studies identifies sex- and ancestry-specific genetic risk loci.

The risk of posttraumatic stress disorder (PTSD) following trauma is heritable, but robust common variants have yet to be identified. In a multi-ethnic cohort including over 30,000 PTSD cases and 170,000 controls we conduct a genome-wide association study of PTSD. We demonstrate SNP-based heritability estimates of 5-20%, varying by sex. Three genome-wide significant loci are identified, 2 in European and 1 in African-ancestry analyses. Analyses stratified by sex implicate 3 additional loci in men. Along with other novel genes and non-coding RNAs, a Parkinson's disease gene involved in dopamine regulation, PARK2, is associated with PTSD. Finally, we demonstrate that polygenic risk for PTSD is significantly predictive of re-experiencing symptoms in the Million Veteran Program dataset, although specific loci did not replicate. These results demonstrate the role of genetic variation in the biology of risk for PTSD and highlight the necessity of conducting sex-stratified analyses and expanding GWAS beyond European ancestry populations

The association between genetically determined ABO blood types and major depressive disorder

ABO blood types and their corresponding antigens have long been assumed to be related to different human diseases. So far, smaller studies on the relationship between mental disorders and blood types yielded contra-dicting results. In this study we analyzed the association between ABO blood types and lifetime major depressive disorder (MDD). We performed a pooled analysis with data from 26 cohorts that are part of the MDD working group of the Psychiatric Genomics Consortium (PGC). The dataset included 37,208 individuals of largely Eu-ropean ancestry of which 41.6% were diagnosed with lifetime MDD. ABO blood types were identified using three single nucleotide polymorphisms in the ABO gene: rs505922, rs8176746 and rs8176747. Regression analyses were performed to assess associations between the individual ABO blood types and MDD diagnosis as well as putative interaction effects with sex. The models were adjusted for sex, cohort and the first ten genetic principal components. The percentage of blood type A was slightly lower in cases than controls while blood type O was more prominent in cases. However, these differences were not statistically significant. Our analyses found no evidence of an association between ABO blood types and major depressive disorder

How is Perceived Community Cohesion and Membership in Community Groups Associated with Children’s Dietary Adequacy in Disadvantaged Communities? A Case of the Indian Sundarbans

Background: Membership in community groups and a sense of community cohesion may facilitate collective action in mobilizing resources towards better health outcomes. This paper explores the relationship of these factors, along with individual level socio-economic variables, to dietary adequacy among children below 6 years of age, a proximate determinant of child malnutrition. Methods: We conducted a cross-sectional survey in Patharpratima block of the Sundarbans in West Bengal, India, using a two-stage, 30 cluster random sampling design. In 1200 sampled households, we used a structured questionnaire to interview mothers of children below 6 years of age on their child’s nutritional intake. We also interviewed household heads to assess perceived community cohesion using a nine item scale, membership in any community self-help organization, and other socio-economic determinants. We used a logistic regression model to assess their association with a minimum acceptable diet among children between 6 months to 6 years. Results: Only 9.33 % children between 6 and 71 months of age received a minimum acceptable diet. With each increase in the perceived community cohesion score (scale 0-9), a child is 1.31 times more likely to have minimum acceptable diet (95 % CI 1.14, 1.50). The odds of minimum acceptable diet were also higher among children whose mothers had primary education (2.09, 95 % CI 1.03, 2.94) as compared to illiterate mothers and in households with surplus food resources (2.72, 95 % CI 1.32, 5.58) as compared to those without surplus or deficit. In contrast, registering at an Anganwadi (government early child development) centre (odds ratio 1.34 95 % CI 0.69, 2.60) and community membership (odds ratio 0.93, 95 % CI 0.59, 1.46) were not associated with minimum acceptable diet. Conclusion: The results are consistent with what is known about the importance of maternal education and access to food resources in ensuring that children have a minimum acceptable diet. Perceived community cohesion seems to play a positive role in children’s diets. Further research needs to clarify which community characteristics and services are the most relevant, how they can better support children’s diets, and how interventions can strengthen these community characteristics and services