Lipid Vesicles Loaded with an HIV-1 Fusion Inhibitor Peptide as a Potential Microbicide

The effective use of fusion inhibitor peptides against cervical and colorectal infections requires the development of sustained release formulations. In this work we comparatively study two different formulations based on polymeric nanoparticles and lipid vesicles to propose a suitable delivery nanosystem for releasing an HIV-1 fusion inhibitor peptide in vaginal mucosa. Polymeric nanoparticles of poly-d,l-lactic-co-glycolic acid (PLGA) and lipid large unilamellar vesicles loaded with the inhibitor peptide were prepared. Both formulations showed average sizes and polydispersity index values corresponding to monodisperse systems appropriate for vaginal permeation. High entrapment efficiency of the inhibitor peptide was achieved in lipid vesicles, which was probably due to the peptide's hydrophobic nature. In addition, both nanocarriers remained stable after two weeks stored at 4 °C. While PLGA nanoparticles (NPs) did not show any delay in peptide release, lipid vesicles demonstrated favorably prolonged release of the peptide. Lipid vesicles were shown to improve the retention of the peptide on ex vivo vaginal tissue in a concentration sufficient to exert its pharmacological effect. Thus, the small size of lipid vesicles, their lipid-based composition as well as their ability to enhance peptide penetration on vaginal tissue led us to consider this formulation as a better nanosystem than polymeric nanoparticles for the sustained delivery of the HIV-1 fusion inhibitor peptide in vaginal tissues

The EGR2 gene is involved in axonal Charcot-Marie-Tooth disease

[EN] Background and purpose: A three-generation family affected by axonal Charcot-Marie-Tooth disease (CMT) was investigated with the aim of discovering genetic defects and to further characterize the phenotype. Methods: The clinical, nerve conduction studies and muscle magnetic resonance images of the patients were reviewed. A whole exome sequencing was performed and the changes were investigated by genetic studies, in silico analysis and luciferase reporter assays. Results: A novel c.1226G>A change (p.R409Q) in the EGR2 gene was identified. Patients presented with a typical, late-onset axonal CMT phenotype with variable severity that was confirmed in the ancillary tests. The in silico studies showed that the residue R409 is an evolutionary conserved amino acid. The p.R409Q mutation, which is predicted as probably damaging, would alter the conformation of the protein slightly and would cause a decrease of gene expression. Conclusions: This is the first report of an EGR2 mutation presenting as an axonal CMT phenotype with variable severity. This study broadens the phenotype of the EGR2-related neuropathies and suggests that the genetic testing of patients suffering from axonal CMT should include the EGR2 gene.This collaborative joint project is awarded by IRDiRC and funded by the Instituto de Salud Carlos III (ISCIII) - Subdireccion General de Evaluacion y Fomento de la Investigacion within the framework of the National R+D+I Plan (Grants IR11/TREAT-CMT, PI12/00946 and PI12/00453), co-funded with FEDER funds. C.E. has a "Miguel Servet' contract funded by the ISCIII and Centro de Investigacion Principe Felipe (CIPF) (Grant no. CPII14/00002). We are also grateful to Itziar Llopis for sample management.Sevilla, T.; Sivera, R.; Martínez-Rubio, D.; Lupo, V.; Chumillas, M.; Calpena-Corpas, E.; Dopazo, J.... (2015). The EGR2 gene is involved in axonal Charcot-Marie-Tooth disease. European Journal of Neurology. 22(12):1548-1555. https://doi.org/10.1111/ene.1278215481555221

MEGARA focal plane subsystems

MEGARA (Multi-Espectrografo en GTC de Alta Resolucion para Astronomia) is the future optical Integral-Field Unit (IFU) and Multi-Object Spectrograph (MOS) for GTC. The Fiber Units are placed at one Folded Cassegrain focus and feed the spectrograph located on a Nasmyth-type platform. This paper summarizes the status of the design of the MEGARA Folded Cassegrain Subsystems after the PDR (held on March 2012), as well as the prototyping that has been carried out during this phase. The MEGARA Fiber Unit has two IFUs: a Large Compact Bundle covering 12.5 arcsec x 11.3 arcsec on sky (100 microns fiber-core), and a Small Compact Bundle, of 8.5 arcsec x 6.7 arcsec (70 microns fiber-core), plus a Fiber MOS positioner, able to place up to 100 mini-bundles 7 fibers each (100 microns fiber-core) in MOS configuration within a 3.5arcmin x 3.5arcmin FOV. A field lens provides a telecentric focal plane where the fibers are located. Microlens arrays couple the telescope beam to the collimator focal ratio at the entrance of the fibers (providing the f/17 to f/3 focal ratio reduction to enter into the fibers). Finally, the fibers, organized in bundles, end in the pseudo-slit plate, which will be placed at the entrance focal plane of the MEGARA spectrographs

MEGARA. Large pupil element tests and performance

MEGARA is a third generation spectrograph for the Spanish 10.4m telescope (GTC) providing two observing modes: a large central Integral Field Unit (IFU), called the Large Compact Bundle (LCB), covering a FOV of 12.5 × 11.3 arcsec^(2) , and a Multi-Object Spectrograph (MOS) with a FOV of 3.5 × 3.5 arcmin^(2) . MEGARA will observe the whole visible range from 3650A to 10000A allowing different spectral resolutions (low, medium and high) with R = 6000, 11000 and 18000 respectively. The dispersive elements are placed at the spectrograph pupil position in the path of the collimated beam and they are composed of a set of volume phase hologram gratings (VPHs) sandwiched between two flat windows and coupled in addition to two prisms in the case of the medium- and high-resolution units. We will describe the tests and setups developed to check the requirements of all units, as well as the obtained performance at laboratory

MEGARA-GTC stellar spectral library: I

MEGARA (Multi Espectrografo en GTC de Alta Resolucion para Astronomia) is an optical (3650-9750 Å), fibre-fed, medium-high spectral resolution (R = 6000, 12 000 and 20 000) instrument for the Gran Telescopio CANARIAS (GTC) 10.4-m telescope, commissioned in the summer of 2017, and currently in operation. The scientific exploitation of MEGARA requires a stellar spectra library to interpret galaxy data and to estimate the contribution of the stellar populations. In this paper, we introduce the MEGARA-GTC spectral library, detailing the rationale behind the building of this catalogue. We present the spectra of 97 stars (21 individual stars and 56 members of the globular cluster M15, which are both subsamples taken during the commissioning runs, and 20 stars from our ongoing GTC Open-Time programme). The spectra have R = 20 000 in the HR-R and HR-I set-ups, centred at 6563 and 8633 Å, respectively. We describe the procedures to reduce and analyse the data. Then, we determine the best-fitting theoretical models to each spectrum through a χ^(2) minimization technique, to derive the stellar physical parameters, and we discuss the results. We have also measured some absorption lines and indices. Finally, we introduce our project to complete the library and the data base in order to make the spectra available to the community

Mapping the ionized gas of the metal-poor HII galaxy PHL 293B with MEGARA

Here we report the first spatially resolved spectroscopic study for the galaxy PHL293B using the high-resolution GTC/MEGARA IFU. PHL293B is a local, extremely metal-poor, high ionization galaxy. This makes PHL 293B an excellent analogue for galaxies in the early Universe. The MEGARA aperture (~12.5''x 11.3'') covers the entire PHL 293B main body and its far-reaching ionized gas. We created and discussed maps of all relevant emission lines, line ratios and physical-chemical properties of the ionized ISM. The narrow emission gas appears to be ionized mainly by massive stars according to the observed diganostic line ratios, regardless of the position across the MEGARA aperture. We detected low intensity broad emission components and blueshifted absorptions in the Balmer lines (H$\alpha$,H$\beta$) which are located in the brightest zone of the galaxy ISM. A chemically homogeneity, across hundreds of parsecs, is observed in O/H. We take the oxygen abundance 12+log(O/H)=7.64 $\pm$ 0.06 derived from the PHL293B integrated spectrum as the representative metallicity for the galaxy. Our IFU data reveal for the first time that the nebular HeII4686 emission from PHL 293B is spatially extended and coincident with the ionizing stellar cluster, and allow us to compute its absolute HeII ionizing photon flux. Wolf-Rayet bumps are not detected excluding therefore Wolf-Rayet stars as the main HeII excitation source. The origin of the nebular HeII4686 is discussed.Comment: 14 pages, 9 Figures, 3 Tables; Accepted for publication in MNRA

Gel-Dispersed Nanostructures Lipid Carriers Loading Thymol Designed for Dermal Pathologies

Purpose: Acne vulgaris is one of the most prevalent dermal disorders affecting skin health and appearance. To date, there is no effective cure for this pathology, and the majority of marketed formulations eliminate both healthy and pathological microbiota. Therefore, hereby we propose the encapsulation of an antimicrobial natural compound (thymol) loaded into lipid nanostructured systems to be topically used against acne.Methods: To address this issue, nanostructured lipid carriers (NLC) capable of encapsulating thymol, a natural compound used for the treatment of acne vulgaris, were developed either using ultrasonication probe or high-pressure homogenization and optimized using 22-star factorial design by analyzing the effect of NLC composition on their physicochemical parameters. These NLC were optimized using a design of experiments approach and were characterized using different physicochemical techniques. Moreover, short-term stability and cell viability using HaCat cells were assessed. Antimicrobial efficacy of the developed NLC was assessed in vitro and ex vivo.Results: NLC encapsulating thymol were developed and optimized and demonstrated a prolonged thymol release. The formulation was dispersed in gels and a screening of several gels was carried out by studying their rheological properties and their skin retention abilities. From them, carbomer demonstrated the capacity to be highly retained in skin tissues, specifically in the epidermis and dermis layers. Moreover, antimicrobial assays against healthy and pathological skin pathogens demonstrated the therapeutic efficacy of thymol-loaded NLC gelling systems since NLC are more efficient in slowly reducing C. acnes viability, but they possess lower antimicrobial activity against S. epidermidis, compared to free thymol.Conclusion: Thymol was successfully loaded into NLC and dispersed in gelling systems, demonstrating that it is a suitable candidate for topical administration against acne vulgaris by eradicating pathogenic bacteria while preserving the healthy skin microbiome.</p

MEGADES: MEGARA Galaxy Discs Evolution Survey. Data Release I: central fields

The main interest of the Science Team for the exploitation of the MEGARA instrument at the 10.4m Gran Telescopio Canarias (GTC hereafter) is devoted to the study of nearby galaxies, with focus on the research of the history of star formation, and chemical and kinematical properties of disc systems. We refer to this project as MEGADES: MEGARA Galaxy Discs Evolution Survey. The initial goal of MEGADES is to provide a detailed study of the inner regions of nearby disc galaxies, both in terms of their spectrophotometric and chemical evolution, and their dynamical characterisation, by disentangling the contribution of in-situ and ex-situ processes to the history of star formation and effective chemical enrichment of these regions. In addition, the dynamical analysis of these inner regions naturally includes the identification and characterization of galactic winds potentially present in these regions. At a later stage, we will extend this study further out in galactocentric distance. The first stage of this project encompasses the analysis of the central regions of a total of 43 nearby galaxies observed with the MEGARA Integral Field Unit for 114 hours, including both Guaranteed Time and Open Time observations. In this paper we provide a set of all the processed data products available to the community and early results from the analysis of these data regarding stellar continuum, ionized and neutral gas features.Comment: Accepted for publication in Astronomy & Astrophysic

SMAD6 variants in craniosynostosis : genotype and phenotype evaluation

PURPOSE: Enrichment of heterozygous missense and truncating SMAD6 variants was previously reported in nonsyndromic sagittal and metopic synostosis, and interaction of SMAD6 variants with a common polymorphism near BMP2 (rs1884302) was proposed to contribute to inconsistent penetrance. We determined the occurrence of SMAD6 variants in all types of craniosynostosis, evaluated the impact of different missense variants on SMAD6 function, and tested independently whether rs1884302 genotype significantly modifies the phenotype. METHODS: We performed resequencing of SMAD6 in 795 unsolved patients with any type of craniosynostosis and genotyped rs1884302 in SMAD6-positive individuals and relatives. We examined the inhibitory activity and stability of SMAD6 missense variants. RESULTS: We found 18 (2.3%) different rare damaging SMAD6 variants, with the highest prevalence in metopic synostosis (5.8%) and an 18.3-fold enrichment of loss-of-function variants comparedwith gnomAD data (P < 10-7). Combined with eight additional variants, ≥20/26 were transmitted from an unaffected parent but rs1884302 genotype did not predict phenotype. CONCLUSION: Pathogenic SMAD6 variants substantially increase the risk of both nonsyndromic and syndromic presentations of craniosynostosis, especially metopic synostosis. Functional analysis is important to evaluate missense variants. Genotyping of rs1884302 is not clinically useful. Mechanisms to explain the remarkable diversity of phenotypes associated with SMAD6 variants remain obscure

A de novo substitution in BCL11B leads to loss of interaction with transcriptional complexes and craniosynostosis

Craniosynostosis, the premature ossification of cranial sutures, is a developmental disorder of the skull vault, occurring in approximately 1 in 2250 births. The causes are heterogeneous, with a monogenic basis identified in ~25% of patients. Using whole-genome sequencing, we identified a novel, de novo variant in BCL11B, c.7C>A, encoding an R3S substitution (p.R3S), in a male patient with coronal suture synostosis. BCL11B is a transcription factor that interacts directly with the nucleosome remodelling and deacetylation complex (NuRD) and polycomb-related complex 2 (PRC2) through the invariant proteins RBBP4 and RBBP7. The p.R3S substitution occurs within a conserved amino-terminal motif (RRKQxxP) of BCL11B and reduces interaction with both transcriptional complexes. Equilibrium binding studies and molecular dynamics simulations show that the p.R3S substitution disrupts ionic coordination between BCL11B and the RBBP4-MTA1 complex, a subassembly of the NuRD complex, and increases the conformational flexibility of Arg-4, Lys-5 and Gln-6 of BCL11B. These alterations collectively reduce the affinity of BCL11B p.R3S for the RBBP4-MTA1 complex by nearly an order of magnitude. We generated a mouse model of the BCL11B p.R3S substitution using a CRISPR-Cas9-based approach, and we report herein that these mice exhibit craniosynostosis of the coronal suture, as well as other cranial sutures. This finding provides strong evidence that the BCL11B p.R3S substitution is causally associated with craniosynostosis and confirms an important role for BCL11B in the maintenance of cranial suture patency