Enhancement of collagen deposition and cross-linking by coupling lysyl oxidase with bone morphogenetic protein-1 and its application in tissue engineering

Cultured cell-derived extracellular matrices (ECM)-based biomaterials exploit the inherent capacity of cells to create highly sophisticated supramolecular assemblies. However, standard cell culture conditions are far from ideal given the fact that the diluted microenvironment does not favor the production of ECM components, a circumstance particularly relevant for collagen. An incomplete conversion of procollagen by C-proteinase/bone morphogenetic protein 1 (BMP1) has been proposed to severely limit in vitro collagen deposition. BMP1 also catalyzes the proteolytic activation of the precursor of the collagen cross-linking enzyme, lysyl oxidase (LOX) to yield the active form, suggesting a deficit in cross-linking activity under standard conditions. We hypothesized that the implementation of fibroblast cultures with LOX and BMP1 may be an effective way to increase collagen deposition. To test it, we have generated stable cell lines overexpressing LOX and BMP1 and studied the effect of supernatants enriched in LOX and BMP1 on collagen synthesis and deposition from fibroblasts. Herein, we demonstrate that the supplementation with LOX and BMP1 strongly increased the deposition of collagen onto the insoluble matrix at the expense of the soluble fraction in the extracellular medium. Using decellularization protocols, we also show that fibroblast-derived matrices regulate adipogenic and osteogenic differentiation of human mesenchymal stem cells (MSC), and this effect was modulated by LOX/BMP1. Collectively, these data demonstrate that we have developed a convenient protocol to enhance the capacity of in vitro cell cultures to deposit collagen in the ECM, representing this approach a promising technology for application in tissue engineeringTis work was supported by grants from Ministerio de Economía y Competitividad (Plan Nacional de I+D+I: SAF2012-34916, and SAF2015-65679-R to F.R-P

Discovery of a deep Seyfert-2 galaxy at z = 0.222 behind NGC 300

We report on the unveiling of the nature of the unidentified X-ray source 3XMM J005450.3-373849 as a Seyfert-2 galaxy located behind the spiral galaxy NGC 300 using Hubble Space Telescope data, new spectroscopic Gemini observations and available XMM-Newton and Chandra data. We show that the X-ray source is positionally coincident with an extended optical source, composed by a marginally resolved nucleus/bulge, surrounded by an elliptical disc-like feature and two symmetrical outer rings. The optical spectrum is typical of a Seyfert-2 galaxy redshifted to z=0.222 +/- 0.001, which confirms that the source is not physically related to NGC 300. At this redshift the source would be located at 909+/-4 Mpc (comoving distance in the standard model). The X-ray spectra of the source are well-fitted by an absorbed power-law model. By tying $N_\mathrm{H}$ between the six available spectra, we found a variable index $\Gamma$ running from ~2 in 2000-2001 years, to 1.4-1.6 in the 2005-2014 period. Alternatively, by tying $\Gamma$, we found variable absorption columns of N_H ~ 0.34 x $10^{-22}$ cm$^{-2}$ in 2000-2001 years, and 0.54-0.75 x $10^{-22}$ cm$^{-2}$ in the 2005-2014 period. Although we cannot distinguish between an spectral or absorption origin, from the derived unabsorbed X-ray fluxes, we are able to assure the presence of long-term X-ray variability. Furthermore, the unabsorbed X-ray luminosities of 0.8-2 x 10$^{43}$ erg s$^{-1}$ derived in the X-ray band are in agreement with a weakly obscured Seyfert-2 AGN at $z \approx 0.22$.Comment: MNRAS, accepte

Analyzing community responses to HIV and AIDS : operational framework and typology

This paper presents a framework for analyzing the community response to HIV and AIDS. On the basis of a review of the literature, six criteria are proposed for characterizing such community responses: (1) the types of organizations and structures implementing the response, (2) the types of activities or services implemented and the beneficiaries of these, (3) the actors involved in and driving community responses, (4) the contextual factors that influence community responses, (5) the extent of community involvement in the response, and (6) the extent to which community responses involve wider partnerships and collaboration.Disability,Civil Society,Community Development and Empowerment,HIV AIDS,Health Monitoring&Evaluation

Thermalization and Cooling of Plasmon-Exciton Polaritons: Towards Quantum Condensation

We present indications of thermalization and cooling of quasi-particles, a precursor for quantum condensation, in a plasmonic nanoparticle array. We investigate a periodic array of metallic nanorods covered by a polymer layer doped with an organic dye at room temperature. Surface lattice resonances of the array---hybridized plasmonic/photonic modes---couple strongly to excitons in the dye, and bosonic quasi-particles which we call plasmon-exciton-polaritons (PEPs) are formed. By increasing the PEP density through optical pumping, we observe thermalization and cooling of the strongly coupled PEP band in the light emission dispersion diagram. For increased pumping, we observe saturation of the strong coupling and emission in a new weakly coupled band, which again shows signatures of thermalization and cooling.Comment: 8 pages, 5 figures including supplemental material. The newest version includes new measurements and corrections to the interpretation of the result

Nigrosine staining of wheat endosperm proteolipid patterns on starch gels

We have previously characterized a group of proteolipids from wheat endosperm, designated CM proteins, which are soluble in chloroformmethanol (2:1, v/v) and have a molecular weight lower than 25,000 daltons (1-3). These have been also studied by Redman and Ewart (4). The CM proteins are suitably fractionated into several components by starch gel electrophoresis at pH 3.2 (1). A sensitive staining procedure was required in connection with genetic studies of these proteins because phenotypes had to be ascertained in small endosperm fractions dissected without impairing normal germination and plant development. We report here on Nigrosine staining conditions for CM proteins under which high sensitivity and selectivity are achieve

Characterization of slow and fast phase nystagmus

A current literature review of the analog and digital process of vestibular and optical kinetic nystagmus reveals little agreement in the methods used by various labs. The strategies for detection of saccade (fast phase velocity component of nystagmus) vary between labs, and most of the process have not been evaluated and validated with a standard database. A survey was made of major vestibular labs in the U.S. that perform computer analyses of vestibular and optokinetic reflexes to stimuli, and a baseline was established from which to standardize data acquisition and analysis programs. The concept of an Error Index was employed as the criterium for evaluating the performance of the vestibular analysis software programs. The performance criterium is based on the detection of saccades and is the average of the percentages of missed detections and false detections. Evaluation of the programs produced results for lateral gaze with saccadic amplitude of one, two, three, five, and ten degrees with various signal-to-noise ratios. In addition, results were obtained for sinusoidal pursuit of 0.05, 0.10, and 0.50 Hz with saccades from one to ten degrees at various signal-to-noise ratios. Selection of the best program was made from the performance in the lateral gaze with three degrees of saccadic amplitude and in the 0.10 Hz sinusoid with three degrees of saccadic amplitude