Rapid simultaneous identification and quantitation of Staphylococcus aureus and Pseudomonas aeruginosa directly from bronchoalveolar lavage specimens using automated microscopy

AbstractDiagnosis of ventilator-assisted pneumonia (VAP) requires pathogen quantitation of respiratory samples. Current quantitative culture methods require overnight growth, and pathogen identification requires an additional step. Automated microscopy can perform rapid simultaneous identification and quantitation of live, surface-immobilized bacteria extracted directly from patient specimens using image data collected over 3 h. Automated microscopy was compared to 1 μL loop culture and standard identification methods for Staphylococcus aureus and Pseudomonas spp. in 53 remnant bronchoalveolar lavage specimens. Microscopy identified 9/9 S. aureus and 7/7 P. aeruginosa in all specimens with content above the VAP diagnostic threshold. Concordance for specimens containing targets above the diagnostic threshold was 13/16, with concordance for sub-diagnostic content of 86/90. Results demonstrated that automated microscopy had higher precision than 1 μL loop culture (range ~0.55 log versus ≥1 log), with a dynamic range of ~4 logs (~103 to 106 CFU/mL)

Generation of Connective Tissue-Free Microvascular Fragment Isolates from Subcutaneous Fat Tissue of Obese Mice

BACKGROUND: Microvascular fragment (MVF) isolates are generated by short-term enzymatic digestion of adipose tissue and contain numerous vessel segments for the vascularization of tissue defects. Recent findings indicate that the functionality of these isolates is determined by the quality of the fat source. Therefore, we compared MVF isolates from subcutaneous adipose tissue of obese and lean mice. METHODS: MVF isolates were generated from subcutaneous adipose tissue of donor mice, which received a high fat or control diet for 12 weeks. The isolates were analyzed in vitro and in vivo. RESULTS: Feeding of mice with a high fat diet induced obesity with adipocyte hypertrophy, resulting in a significantly lower collagen fraction and microvessel density within the subcutaneous fat depots when compared to lean controls. Accordingly, MVF isolates from obese mice also contained a reduced number of MVF per mL adipose tissue. However, these MVF tended to be longer and, in contrast to MVF from lean mice, were not contaminated with collagen fibers. Hence, they could be freely seeded onto collagen-glycosaminoglycan scaffolds, whereas MVF from lean controls were trapped in between large amounts of collagen fibers that clogged the pores of the scaffolds. In line with these results, scaffolds seeded with MVF isolates from obese mice exhibited a significantly improved in vivo vascularization after implantation into full-thickness skin defects. CONCLUSION: Subcutaneous adipose tissue from obese mice facilitates the generation of connective tissue-free MVF isolates. Translated to clinical conditions, these findings suggest that particularly obese patients may benefit from MVFbased vascularization strategies

Ultrahigh-temperature granulite-facies metamorphism and exhumation of deep crust in a migmatite dome during late- to post-orogenic collapse and extension in the central Adirondack Highlands (New York, USA)

This study combines field observations, mineral and whole-rock geochemistry, phase equilibrium modeling, and U-Pb sensitive high-resolution ion microprobe (SHRIMP) zircon geochronology to investigate sillimanite-bearing felsic migmatites exposed on Ledge Mountain in the central Adirondack Highlands (New York, USA), part of an extensive belt of mid-crustal rocks comprising the hinterland of the Mesoproterozoic Grenville orogen. Phase equilibrium modeling suggests minimum peak metamorphic conditions of 960-1025 °C and 11-12.5 kbar during the Ottawan orogeny—sig-nificantly higher pressure-temperature conditions than previously determined—followed by a period of near-isothermal decompression, then isobaric cooling. Petrography reveals abundant melt-related microstructures, and pseudosection models show the presence of at least ~15%-30% melt during buoyancy-driven exhumation and decompression. New zircon data document late Ottawan (re)crys-tallization at ca. 1047 ± 5 to 1035 ± 2 Ma following ultrahigh-temperature (UHT) metamorphism and anatexis on the retrograde cooling path. Inherited zircon cores give a mean date of 1136 ± 5 Ma, which suggests derivation of these felsic granulites by partial melting of older igneous rocks. The ferroan, anhydrous character of the granulites is similar to that of the ca. 1050 Ma Lyon Mountain Granite and consistent with origin in a late- to post-Ottawan extensional environment. We present a model for development of a late Ottawan migmatitic gneiss dome in the central Adirondacks that exhumed deep crustal rocks including the Snowy Mountain and Oregon anorthosite massifs with UHT Ledge Mountain migmatites. Recognition of deep crustal meta-plutonic rocks recording UHT metamorphism in a migmatite gneiss dome has significant implications for crustal behavior in this formerly thickened orogen

Multimessenger Bayesian parameter inference of a binary neutron star merger

The combined detection of a binary neutron star merger in both gravitational waves (GWs) and electromagnetic (EM) radiation spanning the entire spectrum – GW170817/AT2017gfo/GRB170817A – marks a breakthrough in the field of multimessenger astronomy. Between the plethora of modelling and observations, the rich synergy that exists among the available data sets creates a unique opportunity to constrain the binary parameters, the equation of state of supranuclear density matter, and the physical processes at work during the kilonova and gamma-ray burst. We report, for the first time, Bayesian parameter estimation combining information from GW170817, AT2017gfo, and GRB170817 to obtain truly multimessenger constraints on the tidal deformability Λ~∈[302, 860], total binary mass M ∈ [2.722, 2.751] M⊙, the radius of a 1.4 solar mass neutron star R∈[11.3, 13.5]km (with additional 0.2 km systematic uncertainty), and an upper bound on the mass ratio of q ≤ 1.27, all at 90 per cent confidence. Our joint novel analysis uses new phenomenological descriptions of the dynamical ejecta, debris disc mass, and remnant black hole properties, all derived from a large suite of numerical relativity simulations

Multimessenger Bayesian parameter inference of a binary neutron star merger

The combined detection of a binary neutron star merger in both gravitational waves (GWs) and electromagnetic (EM) radiation spanning the entire spectrum – GW170817/AT2017gfo/GRB170817A – marks a breakthrough in the field of multimessenger astronomy. Between the plethora of modelling and observations, the rich synergy that exists among the available data sets creates a unique opportunity to constrain the binary parameters, the equation of state of supranuclear density matter, and the physical processes at work during the kilonova and gamma-ray burst. We report, for the first time, Bayesian parameter estimation combining information from GW170817, AT2017gfo, and GRB170817 to obtain truly multimessenger constraints on the tidal deformability Λ~∈[302, 860], total binary mass M ∈ [2.722, 2.751] M⊙, the radius of a 1.4 solar mass neutron star R∈[11.3, 13.5]km (with additional 0.2 km systematic uncertainty), and an upper bound on the mass ratio of q ≤ 1.27, all at 90 per cent confidence. Our joint novel analysis uses new phenomenological descriptions of the dynamical ejecta, debris disc mass, and remnant black hole properties, all derived from a large suite of numerical relativity simulations

Vascularization of Microvascular Fragment Isolates from Visceral and Subcutaneous Adipose Tissue of Mice

Background: Adipose tissue-derived microvascular fragments (MVF) represent effective vascularization units for tissue engineering. Most experimental studies in rodents exclusively use epididymal adipose tissue as a visceral fat source for MVF isolation. However, in future clinical practice, MVF may be rather isolated from liposuctioned subcutaneous fat tissue of patients. Therefore, we herein compared the vascularization characteristics of MVF isolates from visceral and subcutaneous fat tissue of murine origin. Methods: MVF isolates were generated from visceral and subcutaneous fat tissue of donor mice using two different enzymatic procedures. For in vivo analyses, the MVF isolates were seeded onto collagen-glycosaminoglycan scaffolds and implanted into full-thickness skin defects within dorsal skinfold chambers of recipient mice. Results: By means of the two isolation procedures, we isolated a higher number of MVF from visceral fat tissue when compared to subcutaneous fat tissue, while their length distribution, viability and cellular composition were comparable in both groups. Intravital fluorescence microscopy as well as histological and immunohistochemical analyses revealed a significantly reduced vascularization of implanted scaffolds seeded with subcutaneous MVF isolates when compared to implants seeded with visceral MVF isolates. Light and scanning electron microscopy showed that this was due to high amounts of undigested connective tissue within the subcutaneous MVF isolates, which clogged the scaffold pores and prevented the interconnection of individual MVF into new microvascular networks. Conclusion: These findings indicate the need for improved protocols to generate connective tissue-free MVF isolates from subcutaneous fat tissue for future translational studies

Co-Expression of Wild-Type P2X7R with Gln460Arg Variant Alters Receptor Function

The P2X7 receptor is a member of the P2X family of ligand-gated ion channels. A single-nucleotide polymorphism leading to a glutamine (Gln) by arginine (Arg) substitution at codon 460 of the purinergic P2X7 receptor (P2X7R) has been associated with mood disorders. No change in function (loss or gain) has been described for this SNP so far. Here we show that although the P2X7R-Gln460Arg variant per se is not compromised in its function, co-expression of wild-type P2X7R with P2X7R-Gln460Arg impairs receptor function with respect to calcium influx, channel currents and intracellular signaling in vitro. Moreover, co-immunoprecipitation and FRET studies show that the P2X7R-Gln460Arg variant physically interacts with P2X7R-WT. Specific silencing of either the normal or polymorphic variant rescues the heterozygous loss of function phenotype and restores normal function. The described loss of function due to co-expression, unique for mutations in the P2RX7 gene so far, explains the mechanism by which the P2X7R-Gln460Arg variant affects the normal function of the channel and may represent a mechanism of action for other mutations.Fil: Aprile García, Fernando. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Metzger, Michael W.. Max Planck Institute of Psychiatry; AlemaniaFil: Paez Pereda, Marcelo. Max Planck Institute of Psychiatry; AlemaniaFil: Stadler, Herbert. Affectis Pharmaceuticals; AlemaniaFil: Acuña, Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Liberman, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Senin, Sergio Ariel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Gerez, Juan Atilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; ArgentinaFil: Hoijman, Esteban. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Microscopías Avanzadas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Refojo, Damian. Max Planck Institute of Psychiatry; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mitkovski, Mišo. Max Planck Institute of Experimental Medicine; AlemaniaFil: Panhuysen, Markus. Affectis Pharmaceuticals; AlemaniaFil: Stühmer, Walter. Max Planck Institute of Experimental Medicine; AlemaniaFil: Holsboer, Florian. Max Planck Institute of Psychiatry; Alemania. HMNC Brain Health; AlemaniaFil: Deussing, Jan M.. Max Planck Institute of Psychiatry; AlemaniaFil: Arzt, Eduardo Simon. Max Planck Institute of Psychiatry; Alemania. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigación en Biomedicina de Buenos Aires - Instituto Partner de la Sociedad Max Planck; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentin

Carbon-Rich Mira Variables: Radial Velocities and Distances

Optical radial velocities have been measured for 38 C-type Mira variables. These data together with others in the literature are used to study the differences between optical and CO mm observations for C-Miras and the necessary corrections to the optical velocities are derived in order to obtain the true radial velocities of the variables. The difference between absorption and emission line velocities is also examined. A particularly large difference (+30 km\s) is found in the case of the H-alpha emission line. A catalogue is given of 177 C-Miras with estimated distances and radial velocities. The distances are based on bolometric magnitudes derived in Paper I using SAAO observations or (for 60 of the stars) using non-SAAO photometry. In the latter case the necessary transformations to the SAAO system are derived. These data will be used in paper III to study the kinematics of the C-Miras.Comment: 11 pages, accepted for publication in MNRA

Constraints on the neutron star equation of state from AT2017gfo using radiative transfer simulations

The detection of the binary neutron star GW170817 together with the observation of electromagnetic counterparts across the entire spectrum inaugurated a new era of multi-messenger astronomy. In this study we incorporate wavelength-dependent opacities and emissivities calculated from atomic-structure data enabling us to model both the measured lightcurves and spectra of the electromagnetic transient AT2017gfo. Best-fits of the observational data are obtained by Gaussian Process Regression, which allows us to present posterior samples for the kilonova and source properties connected to GW170817. Incorporating constraints obtained from the gravitational wave signal obtained by the LIGO-Virgo Scientific Collaboration, we present a $90\%$ upper bound on the mass ratio $q \lesssim 1.38$ and a lower bound on the tidal deformability of $\tilde{\Lambda} \gtrsim 197$, which rules out sufficiently soft equations of state. Our analysis is a path-finder for more realistic kilonova models and shows how the combination of gravitational wave and electromagnetic measurements allow for stringent constraints on the source parameters and the supranuclear equation of state