Alzheimer's disease-associated P460L variant of EphA1 dysregulates receptor activity and blood-brain barrier function

INTRODUCTION: Genome-wide association studies link susceptibility to late-onset Alzheimer's disease (LOAD) with EphA1. Sequencing identified a non-synonymous substitution P460L as a LOAD risk variant. Other Ephs regulate vascular permeability and immune cell recruitment. We hypothesized that P460L dysregulates EphA1 receptor activity and impacts neuroinflammation. METHODS: EphA1/P460L receptor activity was assayed in isogenic Human Embryonic Kidney (HEK) cells. Soluble EphA1/P460L (sEphA1/sP460L) reverse signaling in brain endothelial cells was assessed by T-cell recruitment and barrier function assays. RESULTS: EphA1 and P460L were expressed in HEK cells, but membrane and soluble P460L were significantly reduced. Ligand engagement induced Y781 phosphorylation of EphA1 but not P460L. sEphA1 primed brain endothelial cells for increased T-cell recruitment; however, sP460L was less effective. sEphA1 decreased the integrity of the brain endothelial barrier, while sP460L had no effect. DISCUSSION: These findings suggest that P460L alters EphA1-dependent forward and reverse signaling, which may impact blood-brain barrier function in LOAD. Highlights: EphA1-dependent reverse signaling controls recruitment of T cells by brain endothelial cells. EphA1-dependent reverse signaling remodels brain endothelial cell contacts. LOAD-associated P460L variant of EphA1 shows reduced membrane expression and reduced ligand responses. LOAD-associated P460L variant of EphA1 fails to reverse signal to brain endothelial cells.Neuroscience and Mental Health Research Institute (NMHRI), Cardiff University and Sir Geraint Evans Cardiovascular Fund, Cardiff University (grant to A.A. and A.J.R.), L.E.T. by UK Dementia Research Institute, Cardiff University, O.R.M. by School of Medicine, Cardiff University, S.S. and C.L.T. by Eli Lilly (grant to J.W. and A.A.), and C.C. by School of Cellular and Molecular Medicine, University of Bristol. The Bioflux 200 was funded by the Leukemia Research Appeal for Wales, and iBright 1500 was funded by the Ser Cymru II programme (grant CU209 to V.K. and Manon Pritch, which was partially funded by Cardiff University and the European Regional Development Fund through the Welsh Government (Grant CU209 to V.K. and Manon Pritchard)

A search for diffuse bands in the circumstellar envelopes of post-AGB stars

In this work we present the results of a systematic search for diffuse bands (DBs, hereafter) in the circumstellar envelopes of a carefully selected sample of post-AGB stars. We concentrated on the analysis of 9 of the DBs most commonly found in the interstellar medium. The strength of these features is determined using high resolution optical spectroscopy and the results obtained are compared with literature data on field stars affected only by interstellar reddening. Based on the weak features observed in the subsample of post-AGB stars dominated by circumstellar reddening we conclude that the carrier(s) of these DBs must not be present in the circumstellar environment of these sources, or at least not under the excitation conditions in which DBs are formed. The conclusion is applicable to all the post-AGB stars studied, irrespective of the dominant chemistry or the spectral type of the star considered. A detailed radial velocity analysis of the features observed in individual sources confirms this result, as the Doppler shifts measured are found to be consistent with an interstellar origin.Comment: Accepted for A&

Comparative study of CH+ and SH+ absorption lines observed towards distant star-forming regions

Aims. The HIFI instrument onboard Herschel has allowed high spectral resolution and sensitive observations of ground-state transi- tions of three molecular ions: the methylidyne cation CH+, its isotopologue 13CH+, and sulfanylium SH+. Because of their unique chemical properties, a comparative analysis of these cations provides essential clues to the link between the chemistry and dynamics of the diffuse interstellar medium. Methods. The CH+, 13CH+, and SH+ lines are observed in absorption towards the distant high-mass star-forming regions (SFRs) DR21(OH), G34.3+0.1, W31C, W33A, W49N, and W51, and towards two sources close to the Galactic centre, SgrB2(N) and SgrA*+50. All sight lines sample the diffuse interstellar matter along pathlengths of several kiloparsecs across the Galactic Plane. In order to compare the velocity structure of each species, the observed line profiles were deconvolved from the hyperfine structure of the SH+ transition and the CH+, 13CH+, and SH+ spectra were independently decomposed into Gaussian velocity components. To analyse the chemical composition of the foreground gas, all spectra were divided, in a second step, into velocity intervals over which the CH+, 13CH+, and SH+ column densities and abundances were derived. Results. SH+ is detected along all observed lines of sight, with a velocity structure close to that of CH+ and 13CH+. The linewidth distributions of the CH+, SH+, and 13CH+ Gaussian components are found to be similar. These distributions have the same mean ( ~ 4.2 km s-1) and standard deviation (\sigma(\delta\u{psion}) ~ 1.5 km s-1). This mean value is also close to that of the linewidth distribution of the CH+ visible transitions detected in the solar neighbourhood. We show that the lack of absorption components narrower than 2 km s-1 is not an artefact caused by noise: the CH+, 13CH+, and SH+ line profiles are therefore statistically broader than those of most species detected in absorption in diffuse interstellar gas (e. g. HCO+, CH, or CN). The SH+/CH+ column density ratio observed in the components located away from the Galactic centre spans two orders of magnitude and correlates with the CH+ abundance. Conversely, the ratio observed in the components close to the Galactic centre varies over less than one order of magnitude with no apparent correlation with the CH+ abundance. The observed dynamical and chemical properties of SH+ and CH+ are proposed to trace the ubiquitous process of turbulent dissipation, in shocks or shears, in the diffuse ISM and the specific environment of the Galactic centre regions

Abundances and Behavior of 12CO, 13CO, and C2 in Translucent Sight Lines

Using UV spectra obtained with FUSE, HST, and/or IUE, we determine interstellar column densities of 12CO, 13CO, and/or C_2 for ten Galactic sight lines with 0.37<E(B-V)<0.72. The N(CO)/N(H_2) ratio varies over a factor of 100 in this sample, due primarily to differences in N(CO). For a given N(H_2), published models of diffuse and translucent clouds predict less CO than is observed. The J=1-3 rotational levels of 12CO are sub-thermally populated in these sight lines, with T_ex typically between 3 and 7 K. In general, there is no significant difference between the excitation temperatures of 12CO and 13CO. Fits to the higher resolution CO line profiles suggest that CO (like CN) is concentrated in relatively cold, dense gas. We obtain C_2 column densities from the F-X (1-0) and (0-0) bands (1314 and 1341 A), the D-X (0-0) band (2313 A), and the A-X (3-0) and (2-0) bands (7719 and 8757 A). Comparisons among those N(C_2) yield a set of mutually consistent f-values for the UV and optical C_2 bands, but also reveal some apparent anomalies within the F-X (0-0) band. Both the kinetic temperature inferred from the C_2 rotational populations (up to J=18) and the excitation temperature T_02(C_2) are generally smaller than the corresponding T_01(H_2). Incorporating additional data for K I, HD, CH, C_2, C_3, CN, and CO from the literature (for a total sample of 74 sight lines), we find that (1) CO is most tightly correlated with CN; (2) the ratios 12CO/H_2 and 13CO/H_2 both are fairly tightly correlated with the density indicator CN/CH (but C_2/H_2 is not); and (3) the ratio 12CO/13CO is somewhat anti-correlated with both CN/CH and N(CO). Sight lines with 12CO/13CO below the average local Galactic value of 12C/13C appear to sample colder, denser gas in which isotope exchange reactions have enhanced 13CO, relative to 12CO.Comment: 78 pages, 24 figures, accepted to ApJ

Measurement of the Bottom-Strange Meson Mixing Phase in the Full CDF Data Set

We report a measurement of the bottom-strange meson mixing phase \beta_s using the time evolution of B0_s -> J/\psi (->\mu+\mu-) \phi (-> K+ K-) decays in which the quark-flavor content of the bottom-strange meson is identified at production. This measurement uses the full data set of proton-antiproton collisions at sqrt(s)= 1.96 TeV collected by the Collider Detector experiment at the Fermilab Tevatron, corresponding to 9.6 fb-1 of integrated luminosity. We report confidence regions in the two-dimensional space of \beta_s and the B0_s decay-width difference \Delta\Gamma_s, and measure \beta_s in [-\pi/2, -1.51] U [-0.06, 0.30] U [1.26, \pi/2] at the 68% confidence level, in agreement with the standard model expectation. Assuming the standard model value of \beta_s, we also determine \Delta\Gamma_s = 0.068 +- 0.026 (stat) +- 0.009 (syst) ps-1 and the mean B0_s lifetime, \tau_s = 1.528 +- 0.019 (stat) +- 0.009 (syst) ps, which are consistent and competitive with determinations by other experiments.Comment: 8 pages, 2 figures, Phys. Rev. Lett 109, 171802 (2012

A microfluidic device with fluorimetric detection for intracellular components analysis

An integrated microfluidic system that coupled lysis of two cell lines: L929 fibroblasts and A549 epithelial cells, with fluorescence-based enzyme assay was developed to determine β-glucocerebrosidase activity. The microdevice fabricated in poly(dimethylsiloxane) consists of three main parts: a chemical cell lysis zone based on the sheath flow geometry, a micromeander and an optical fibers detection zone. Unlike many methods described in literature that are designed to analyse intracellular components, the presented system enables to perform enzyme assays just after cell lysis process. It reduces the effect of proteases released in lysis process on determined enzymes. Glucocerebrosidase activity, the diagnostic marker for Gaucher’s disease, is the most commonly measured in leukocytes and fibroblasts using 4-methylumbelliferyl-β-D-glucopyranoside as synthetic β-glucoside. The enzyme cleavage releases the fluorescent product, i.e. 4-methylumbelliferone, and its fluorescence is measured as a function of time. The method of enzyme activity determination described in this paper was adapted for flow measurements in the microdevice. The curve of the enzymatic reaction advancement was prepared for three reaction times obtained from application of different flow rates of solutions introduced to the microsystem. Afterwards, determined β-glucocerebrosidase activity was recalculated with regard to 105 cells present in samples used for the tests. The obtained results were compared with a cuvette-based measurements. The lysosomal β-glucosidase activities determined in the microsystem were in good correlation with the values determined during macro-scale measurements

Improved Heterosis Prediction by Combining Information on DNA- and Metabolic Markers

Background: Hybrids represent a cornerstone in the success story of breeding programs. The fundamental principle underlying this success is the phenomenon of hybrid vigour, or heterosis. It describes an advantage of the offspring as compared to the two parental lines with respect to parameters such as growth and resistance against abiotic or biotic stress. Dominance, overdominance or epistasis based models are commonly used explanations. Conclusion/Significance: The heterosis level is clearly a function of the combination of the parents used for offspring production. This results in a major challenge for plant breeders, as usually several thousand combinations of parents have to be tested for identifying the best combinations. Thus, any approach to reliably predict heterosis levels based on properties of the parental lines would be highly beneficial for plant breeding. Methodology/Principal Findings: Recently, genetic data have been used to predict heterosis. Here we show that a combination of parental genetic and metabolic markers, identified via feature selection and minimum-description-length based regression methods, significantly improves the prediction of biomass heterosis in resulting offspring. These findings will help furthering our understanding of the molecular basis of heterosis, revealing, for instance, the presence of nonlinear genotype-phenotype relationships. In addition, we describe a possible approach for accelerated selection in plant breeding

Biallelic Variants in PYROXD2 Cause a Severe Infantile Metabolic Disorder Affecting Mitochondrial Function

Pyridine Nucleotide-Disulfide Oxidoreductase Domain 2 (PYROXD2; previously called YueF) is a mitochondrial inner membrane/matrix-residing protein and is reported to regulate mitochondrial function. The clinical importance of PYROXD2 has been unclear, and little is known of the protein’s precise biological function. In the present paper, we report biallelic variants in PYROXD2 identified by genome sequencing in a patient with suspected mitochondrial disease. The child presented with acute neurological deterioration, unresponsive episodes, and extreme metabolic acidosis, and received rapid genomic testing. He died shortly after. Magnetic resonance imaging (MRI) brain imaging showed changes resembling Leigh syndrome, one of the more common childhood mitochondrial neurological diseases. Functional studies in patient fibroblasts showed a heightened sensitivity to mitochondrial metabolic stress and increased mitochondrial superoxide levels. Quantitative proteomic analysis demonstrated decreased levels of subunits of the mitochondrial respiratory chain complex I, and both the small and large subunits of the mitochondrial ribosome, suggesting a mitoribosomal defect. Our findings support the critical role of PYROXD2 in human cells, and suggest that the biallelic PYROXD2 variants are associated with mitochondrial dysfunction, and can plausibly explain the child’s clinical presentation