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

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

On the Reliability of Cross Correlation Function Lag Determinations in Active Galactic Nuclei

Many AGN exhibit a highly variable luminosity. Some AGN also show a pronounced time delay between variations seen in their optical continuum and in their emission lines. In effect, the emission lines are light echoes of the continuum. This light travel-time delay provides a characteristic radius of the region producing the emission lines. The cross correlation function (CCF) is the standard tool used to measure the time lag between the continuum and line variations. For the few well-sampled AGN, the lag ranges from 1-100 days, depending upon which line is used and the luminosity of the AGN. In the best sampled AGN, NGC 5548, the H_beta lag shows year-to-year changes, ranging from about 8.7 days to about 22.9 days over a span of 8 years. In this paper it is demonstrated that, in the context of AGN variability studies, the lag estimate using the CCF is biased too low and subject to a large variance. Thus the year-to-year changes of the measured lag in NGC 5548 do not necessarily imply changes in the AGN structure. The bias and large variance are consequences of finite duration sampling and the dominance of long timescale trends in the light curves, not due to noise or irregular sampling. Lag estimates can be substantially improved by removing low frequency power from the light curves prior to computing the CCF.Comment: To appear in the PASP, vol 111, 1999 Nov; 37 pages; 10 figure

Effects of Osteopathic Visceral Treatment in Patients with Gastroesophageal Reflux: A Randomized Controlled Trial

Osteopathic manual treatment has been recommended as a non-pharmacological therapy for Gastroesophageal Reflux Disease (GERD). However, to date, no study has supported the effectiveness of this intervention with respect to the symptoms of the disease. Our goal was to assess the effect of an osteopathic manual technique for the lower esophageal sphincter on GERD symptoms, cervical mobility and on the C4 spinous process pressure pain threshold (PPTs). Methods: A randomized, double-blind placebo-controlled trial was performed. Sixty subjects suffering from GERD participated in this study and were randomly assigned to either an experimental group (EG) (n = 29), who received the osteopathic technique for the lower esophageal sphincter, or to a control group (CG) (n = 31), who received a manual contact, which mimicked the osteopathic technique without exerting any therapeutic force. Randomization was computer-generated, with allocation concealed by sequentially numbered, opaque, sealed envelopes. The GerdQ questionnaire was used to assess symptom changes the week after intervention. Cervical Range of Motion (CROM) and algometer were used to evaluate cervical mobility and PPTs before and after both treatments. Before-after between groups comparison (t-test) was used for statistical analysis of the outcome, with two measurement points (GerdQ), while repeated-measures ANOVA was used for those outcomes with four measurement points (CROM and PPT). Results: The application of the osteopathic manual treatment in subjects with GERD produced a significant improvement in symptoms one week after the intervention (p = 0.005) with a between-groups difference of 1.49 points in GerdQ score (95% CI: 0.47-2.49). PPT C4 improved in the EG after the treatment (p = 0.034; eta(2) = 0.048) (between-groups difference 8.78 Newton/cm(2); 95% CI: 0.48-17.09). CROM also increased in the EG compared to the CG (p < 0.001; eta(2) = 0.108) (between-groups difference 33.89 degrees; 95% CI: 15.17-52.61). Conclusions: The manual osteopathic technique produces an improvement in GERD symptoms one week after treatment, cervical mobility, and PPTs. This may mean that osteopathic treatment is useful for improving symptoms of GERD

Dynamics of the Lyman alpha and C IV emitting gas in 3C 273

In this paper we study the variability properties of the Lyman alpha and C IV emission lines in 3C273 using archival IUE observations. Our data show for the first time the existence of variability on time scales of several years. We study the spatial distribution and the velocity field of the emitting gas by performing detailed analyses on the line variability using correlations, 1D and 2D response functions, and principal component analysis. In both lines we find evidence for two components, one which has the dynamic properties of gas in Keplerian motion around a black hole with a mass of the order of 10^9 Mo, and one which is characterized by high, blue-shifted velocities at large lag. There is no indication of the presence of optically thick emission medium neither in the Lya, nor in the Civ response functions. The component characterized by blue-shifted velocities, which is comparatively much stronger in Civ than in Lya, is more or less compatible with being the result of gas falling towards the central black hole with free-fall acceleration. We propose however that the line emission at high, blue-shifted velocities is better explained in terms of entrainment of gas clouds by the jet. This gas is therefore probably collisionally excited as a result of heating due to the intense infrared radiation from the jet, which would explain the strength of this component in Civ relative to Lya. This phenomenon might be a signature of disk-jet interaction.Comment: 16 pages, 10 figures. Accepted for publication in ApJ. Uses aaste

Efficacy and safety of intermittent intravenous outpatient administration of levosimendan in patients with advanced heart failure: the LION-HEART multicentre randomised trial

Aims: The LION-HEART study was a multicentre, double-blind, randomised, parallel-group, placebo-controlled trial evaluating the efficacy and safety of intravenous administration of intermittent doses of levosimendan in outpatients with advanced chronic heart failure. Methods and results: Sixty-nine patients from 12 centres were randomly assigned at a 2: 1 ratio to levosimendan or placebo groups, receiving treatment by a 6-hour intravenous infusion (0.2 mu g/kg/min without bolus) every 2weeks for 12weeks. The primary endpoint was the effect on serum concentrations of N-terminal pro-B-type natriuretic peptide (NT-proBNP) throughout the treatment period in comparison with placebo. Secondary endpoints included evaluation of safety, clinical events and health-related quality of life (HRQoL). The area under the curve (AUC, pg.day/mL) of the levels of NT-proBNP over time for patients who received levosimendan was significantly lower than for the placebo group {344 x 10(3) [95% confidence interval (CI) 283 x 10(3)-404 x 10(3)] vs. 535 x 10(3) [443 x 10(3)-626 x 10(3)], P = 0.003}. In comparison with the placebo group, the patients on levosimendan experienced a reduction in the rate of heart failure hospitalisation (hazard ratio 0.25; 95% CI 0.11-0.56; P = 0.001). Patients on levosimendan were less likely to experience a clinically significant decline in HRQoL over time (P = 0.022). Adverse event rates were similar in the two treatment groups. Conclusions: In this small pilot study, intermittent administration of levosimendan to ambulatory patients with advanced systolic heart failure reduced plasma concentrations of NT-proBNP, worsening of HRQoL and hospitalisation for heart failure. The efficacy and safety of this intervention should be confirmed in larger trials