Role of extracellular vesicles in retinitis pigmentosa

Retinitis pigmentosa (RP), an inherited degenerative retinal disease, is associated with progressive photoreceptor degeneration, which leads eventually to blindness. To date, the precise mechanisms leading to cell death remain unknown and no adequate treatment is available. Poly ADP ribose polymerase (PARP) over activity is involved in photoreceptor degeneration and, in mice models, its pharmacological inhibition protects the retina. Additionally, retinal cell survival depends of adequate reception and processing of the information and appropriate cellular communication. Initially, the extracellular vesicles (EVs) were recognized as a mechanism for discharging useless cellular components. Growing evidence has elucidated their roles in cell–cell communication by carrying nucleic acids, proteins, and lipids that can, in turn, regulate behavior of target cells. Nevertheless, the role of EVs in blinding diseases, such a RP, is far from being understood. The present project aims to investigate the EVs implication in retinal degeneration, including their release and cargo, their influence in neighboring cells, and the relationship with PARP activity in the retina. Rd1 and Rd10 mice - two well-known animal model for RP, which hold a mutation in the beta subunit of the phosphodiesterase 6 gene (PDE6) – helped advanced the understanding of the retinal degeneration. Section from rd1 and rd10 mice and organotypic retinal explants from rd10 were used to investigate cellular communication by EVs. CD9 and CD81 tetraspanins were studied to investigate EVs activity at tissue level by immunostaining. Inhibition of PARP activity was performed using Olaparib. Immunohistochemistry was carried out to evaluate PARylated proteins and immunostaining was performed to determinate rhodopsin (rho) expression, Müller glia cell activity, and cyclic guanosine monophosphate (cGMP) levels after olaparib treatment. Also, inmunofluorescence was used to study EVs and their colocalization with cilia in rd10 retinae after PARP inhibition. EVs were isolated using ultrafiltration and size exclusion chromatography or a commercial isolation kit, depending on downstream applications. Nanosight analysis, electron microscope, Fluorescence-Activated Cell Sorting (FACs), dot blot, and proteomics were used to characterize the EVs. Moreover, rd10 retinas were treated with EV from wt and vice-versa. Inmunostaining assays against CD9, CD81, rho, and IBA-1 (microglia marker) were carried out after EVs treatments. TUNEL assay was used to evaluate cell viability, thickness, and row photoreceptor number in the outer nuclear layer (ONL) after Olaparib and EVs treatments. EVs release changes with the age in wt mice and also under retinal degeneration in rd1 and rd10 in different retinal layers. PARP inhibition by Olaparib rescues photoreceptors and also modify the EVs release and cargo in rd10 mice. The EVs release was increased in rd10 retinae and the protein cargo was modified under retinal degeneration. Moreover, EVs from rd10 retinae had the ability to damage wt retinas and something similar was produced after treated rd10 retinae with EVs from wt. This data strongly suggests the implication of EVs in retina development and degeneration.Retinitis pigmentosa (RP) es una enfermedad hereditaria de la retina que produce degeneración progresiva de los fotorreceptores, generando ceguera. Hasta la fecha, los mecanismos precisos que conducen a la muerte celular son desconocidos y no existe un tratamiento adecuado. La sobreactividad de la Poli ADP-ribosa polimerasa (PARP) se encuentra implicada en la degeneración de los fotorreceptores y, en modelos animales, se ha observado que inhibición de PARP protege la retina. Además, la supervivencia celular de la retina depende de una adecuada recepción y procesamiento de la información y de una apropiada comunicación celular. Inicialmente, las vesículas extracelulares (VEs) fueron descritas como un mecanismo para eliminar componentes celulares inútiles. La creciente evidencia ha aclarado su papel en la comunicación celular, transportando ácidos nucleicos, proteínas y lípidos que pueden, a su vez, regular el comportamiento de las células diana. Sin embargo, el papel de las VEs en las enfermedades que cursan con ceguera ceguera, como la RP, está lejos de ser entendido. El presente proyecto tiene como objetivo investigar la implicación de las VEs en la degeneración de la retina, incluida su liberación y carga, su influencia en las células vecinas y la relación con la actividad de PARP. Los ratones Rd1 y Rd10, dos modelos animales de RP, que contienen una mutación en la subunidad beta del gen de la fosfodiesterasa 6 (PDE6), han sido de gran ayuda para entender como degenera la retina en la RP. Cortes de ratones rd1 y rd10 y cultivos organotípicos de retina de ratones rd10 se usaron para investigar la comunicación celular mediante VEs. Se estudió a expresión de las tetraspaninas CD9 y CD81 para investigar la actividad de las VEs a nivel retiniano. Para inhibir la actividad de PARP se realizó usando Olaparib. La inmunohistoquímica se llevó a cabo para evaluar las proteínas PARiladas y la inmunotinción se empleó para determinar la expresión de rodopsina (rho), la actividad de las células de Müller y el nivel de guanosín monofosfato cíclico (GMPc) después del tratamiento con olaparib. Además, se utilizó inmunofluorescencia para estudiar las VEs y su colocalización con cilios en retinas rd10, tras la inhibición de PARP. Las VEs fueron aisladas mediante ultrafiltración y cromatografía de exclusión por tamaño o con un kit de aislamiento comercial, dependiendo de las aplicaciones posteriores. Para caracterizar las VEs se utilizó la técnica Nanosight, microscopía electrónica, citometría de flujo (FACs), dot blot y proteómica. Además, las retinas rd10 se trataron con VEs de wt y viceversa. La inmunotinción frente a CD9, CD81, rho e IBA-1 (marcador de microglia) se llevó a cabo después de los tratamientos con VEs. El ensayo TUNEL se utilizó para evaluar la viabilidad celular, el grosor y el número de fotorreceptores en la capa nuclear externa después de los tratamientos con Olaparib y VEs. La liberación de VEs cambia con la edad en ratones wt y también bajo degeneración retiniana en rd1 y rd10 en diferentes capas retinianas. La inhibición de PARP por Olaparib rescata fotorreceptores y también modifica la liberación y carga de las VEs en ratones rd10. La liberación de VEs aumentó en las retinas rd10 y el cargo proteico se modificó con la degeneración de la retina. Además, las VEs de las retinas rd10 presentaron la capacidad de dañar las retinas wt y se produjo algo similar después de que las retinas rd10 fueran tratadas con las VEs de wt. Estos datos sugieren la implicación de las VEs en el desarrollo y la degeneración de la retina.MedicinaCiencias de la Salu

Co-term and social networks analysis for the generation of subject maps

La mejora de las capacidades de búsqueda y de las interfaces de los opacs sigue siendo uno de los principales desafíos para las bibliotecas, especialmente en lo que respecta al acceso por materias. Las interfaces visuales pueden facilitar la recuperación. El objetivo del presente trabajo es explorar si la combinación de técnicas de análisis de co-términos y de redes sociales resulta ser una metodología válida para la generación de mapas temáticos de la colección. La principal conclusión es que el método es válido, y que los mapas obtenidos podrían servir como interfaz visual para el acceso por materias. También es útil para detectar problemas en los criterios de indización y contribuir a la mejora de la calidad de la descripción temática del conjunto documentalImprovement of the interfaces and search capacities of the opacs continues to be one of the main challenges for libraries, especially with regard to searching by subject. The visual interfaces can help with retrieval. This paper explores the combination of co-term analysis techniques with social networks for the generation of subject maps of collections. The main conclusion is that the methodology applied is valid, and that the maps could be used for browsing and subject retrieval. It would also be useful for improving the quality of indexingFil: Miguel, Sandra. Universidad Nacional de La Plata. Facultad de Humanidades y Ciencias de la Educación; Argentina

Zeolite/iron oxide composite as sorbent for magnetic solid-phase extraction of benzene, toluene, ethylbenzene and xylenes from water samples prior to gas chromatography-mass spectrometry

This study reports a new composite based on ZSM-5 zeolite decorated with iron oxide magnetic nanoparticles as a valuable sorbent for magnetic solid-phase extraction (MSPE). A proposal is made to determine benzene, toluene, ethylbenzene and xylenes (BTEX) as model analytes in water samples using gas chromatography-mass spectrometry. A two-step multivariate optimization strategy, using Plackett⬜Burman and circumscribed central composite designs, was employed to optimize experimental parameters affecting MSPE. The method was evaluated under optimized extraction conditions (i.e., amount of sorbent, 138 mg; extraction time, 11 min; sample pH, pH of water (i.e., 5.5⬜6.5); eluent solvent volume, 0.5 mL; and elution time, 5 min), obtaining a linear response from 1 to 100 μg L↙1 for benzene; from 10 to 100 μg L↙1 for toluene, ethylbenzene and o-xylene; and from 10 to 75 μg L↙1 for m,p-xylene. The repeatability of the proposed method was evaluated at a 40 μg L↙1 spiking level and coefficients of variation ranged between 8 and 11% (n = 5). Limits of detection were found to be 0.3 μg L↙1 for benzene and 3 μg L↙1 for the other analytes. These values satisfy the current normative of the Environmental Protection Agency and European Union for BTEX content in waters for human consumption. Finally, drinking water, wastewater and river water were selected as real water samples to assess the applicability of the method. Relative recoveries varied between 85% and 114% showing negligible matrix effects.The authors would like to thank the Spanish Ministry of Science and Innovation (project no. CTQ2011-23968), Generalitat Valenciana (Spain) (projects nos. GVA/2014/096 and PROMETEO/2013/038) for the financial support. E. Fernández also thanks Ministry of Education for her FPU grant (FPU13/03125)

Determination of siloxanes in water samples employing graphene oxide/Fe3O4 nanocomposite as sorbent for magnetic solid‐phase extraction prior to GC–MS

A new, fast, simple, and environmentally friendly analytical method has been developed to determine six siloxanes in water samples: octamethyltrisiloxane, octamethylcyclotetrasiloxane, decamethyltetrasiloxane, decamethylcyclopentasiloxane, dodecamethylpentasiloxane and dodecamethylcyclohexasiloxane. The analytical method consists of magnetic solid‐phase extraction employing graphene oxide/Fe3O4 as sorbent for the separation and preconcentration of siloxanes prior to GC–MS determination. The extraction procedure was optimized by means of a Plackett‐Burman design. Under the optimized extraction conditions (graphene oxide/Fe3O4, 20 mg; extraction time, 10 min; eluent volume, 0.5 mL ACN; elution time, 2.5 min; sample volume, 20 mL), the method rendered repeatability levels with a relative standard deviation between 9 and 20% (n = 6, 10 μg/L). Methodological limits of detection ranged from 0.003 to 0.1 μg/L. The linearity of the method was studied between the methodological limit of quantification and 100 μg/L, obtaining correlation coefficient values between 0.990 and 0.999. The applicability of the method was assessed by analyzing drinking, river and wastewater samples. Relative recovery values ranged between 70 and 120% (1 and 60 μg/L spiking level) showing that the matrix had a negligible effect on extraction. Finally, the greenness of this method was confirmed by the semiquantitative Eco‐Scale metrics.The authors would like to thank “Vicerrectorado de Investigación y Transferencia de Conocimiento” of the University of Alicante (UAUSTI16-04), the Ministry of Science and Innovation of Spain (project no. CTQ2011-23968), the Ministry of Economy, Industry and Competitiveness of Spain (project no. CTQ2016-79991-R, AEI/FEDER, UE) for the financial support. L. Costa thanks the Capes Foundation within the Ministry of Education in Brazil (Process 99999.012013/2013-07)

Factors associated with early childhood caries in Chile

AbstractObjectiveTo determine the prevalence of caries and identify the associated factors among children aged 2 and 4 years in southern Chile.MethodsA cross-sectional study was conducted in a random sample of 2,987 children. Dental examinations were performed in all participating children and a self-administered survey was administered to their primary caregivers. The statistical analysis included a zero inflated negative binomial regression model.ResultsThe prevalence of caries was 20.3% for 2-year-olds and 52.7% for 4-year-olds. The factors associated with caries included the following: living in the Bio-Bío region, PR2year-old: 1.65 (1.10-2.47), PR4year-old: 1.44 (1.18-1.75) and having a low socioeconomic position, PR2year-old: 5.39 (2.14-13.57), PR4year-old: 3.71 (2.68-5.13) or a medium socioeconomic position, PR2year-old: 2.79 (1.06-7.33), PR4year-old: 2.70 (1.92-3.79). In 4-year-olds, a high frequency of consuming sugary drinks at bedtime (PR: 1.30; 1.06-1.59), not brushing teeth daily (PR: 1.37; 1.18-1.60), and the presence of gingivitis (PR: 1.64; 1.28-2.09) were also associated with caries.ConclusionsThe high prevalence of caries in this young group is concerning. The factors associated with caries identified in this study could help in the design of preventive interventions at early ages

Determination of four bisphenols in water and urine samples by magnetic dispersive solid‐phase extraction using a modified zeolite/iron oxide composite prior to liquid chromatography diode array detection

A novel approach is presented to determine four bisphenols in water and urine samples, employing magnetic dispersive solid‐phase extraction combined with liquid chromatography and diode array detection. A modified zeolite‐based magnetic composite was used as an efficient sorbent, combining the advantages of magnetic materials with the remarkable properties of zeolites. A multivariate optimization design was employed to optimize some experimental factors affecting magnetic dispersive solid‐phase extraction. The method was evaluated under optimized conditions (i.e., amount of sorbent, 50 mg; sample pH, unadjusted; NaCl concentration, 1.25%; extraction and elution time, 2 min; eluent solvent, ethanol; eluent solvent volume, 400 µL), obtaining good linearity with correlation coefficients ranging between 0.995 and 0.999 (N = 5) (from 2 to 250 µg/L for bisphenol A, bisphenol AP, and bisphenol P and from 5 to 250 µg/L for bisphenol AF). Method repeatability was assessed obtaining coefficients of variation between 3 and 11% (n = 6). Finally, the method was applied to spiked real samples, obtaining for water samples relative recoveries between 83 and 105%, and for urine samples between 81 and 108% for bisphenol A, bisphenol AP, and bisphenol AF, and between 47 and 59% for bisphenol P.The authors would like to thank the Spanish Ministry of Economy and Competitiveness and European Union (FEDER funds) (project no. CTQ2016-79991-R) and Generalitat Valenciana (Spain) (project no. PROMETEO/2018/087) for the financial support. P. Baile also thanks Ministry of Education, Culture and Sports for her FPU grant (FPU14/04589)

Prevención de úlceras por presión relacionadas con la posición prono en pacientes con COVID-19 en la unidad de cuidados intensivos. Una revisión integrativa

Objetivo: Determinar qué actividades y/o intervenciones de enfermería pueden llevarse a cabo para prevenir la aparición de úlceras por presión relacionadas con la posición prono en pacientes con Covid-19 en la Unidad de Cuidados Intensivos. Metodología: Se ha realizado una búsqueda bibliográfica en las bases de datos MEDLINE (a través de PubMed), Cochrane, LILACS, Web Of Science (WOS) y SCOPUS. Para la selección de los artículos incluidos en la revisión se han tenido en cuenta los criterios de selección determinados en la revisión. Se han realizado 4 estrategias de búsqueda diferentes desde lo más concreto a lo más general para así obtener estudios relevantes centrados en el tema de interés de nuestra revisión. Resultados: Finalmente se han seleccionado 11 artículos entre ellos revisiones integrativas, revisiones literarias, estudios de cohortes, estudios quasi-experimentales antes-después, planes de cuidados y hoja de recomendaciones. Se ha evaluado la calidad de los artículos a través de diferentes check-list dependiendo del diseño de cada estudio localizados en Equator Network. Conclusión: Las medidas que se llevarán a cabo para prevenir UPP son: la valoración del riesgo mediante escalas validadas y evaluando regularmente la integridad cutánea, realizar cuidados de la piel así como higiene adecuada de la misma, aplicar apósitos profilácticos en prominencias y zonas de riesgo, manejo de la redistribución de la presión, colocar al paciente en posición del nadador así como la formación continuada de las enfermeras de la UCI y contar con una enfermera especialista en cuidados de las heridas

Edificio de 7 viviendas, garaje y trasteros

[ES] En el presente proyecto se describe, analiza y evalúa el seguimiento de la ejecución de un edificio de 7 viviendas garaje y trasteros en Albacete. Para poder llevarlo a cabo, se han hecho visitas diarias a la obra, mantenido contacto con la empresa promotora y he trabajado con la empresa constructora. Toda la documentación aportada y estudiada en el TGF ha sido facilitada por la empresa constructora donde realizaba las prácticas y desde donde realicé el estudio del proyecto de ejecución de la obra. En este proyecto se trabajan los distintos ámbitos que podemos encontrar en el oficio de arquitecto técnico: el estudio y análisis del proyecto, donde se han comparado todos los documentos que lo componen y corroborando que cumple el CTE; el diario de obra, realizando visitas a obra; el seguimiento de calidad, cuidando la correcta ejecución; control y seguimiento de la seguridad y salud, se comprueba el uso de las medidas de seguridad; y el seguimiento económico, en el cual se comprueba el seguimiento de la programación. Con todo ello se consigue una visión general de la gestión integral de una construcción.[EN] This project describes, analyzes and evaluates the monitoring of the execution of a building with 7 garage dwellings and storage rooms in Albacete. In order to carry it out, daily visits to the site have been made, I was in contact with the promoter company and I worked with the construction company. All the documentation contributed and studied in the TGF has been facilitated by the construction company where I carried out the internship and from where I made the study of the execution project of the work. In this project the different areas that we can find in the technical architect office are studied: the study and analysis of the project, where all the documents that compose it have been compared and corroborating that the CTE complies; the daily work, making visits to work; Quality monitoring, taking care of the correct execution; control and monitoring of safety and health, the use of safety measures is checked; and the economic follow-up, in which the monitoring of the programming is checked. With all this we get an overview of the integral management of a construction.González Vidal, L. (2018). Edificio de 7 viviendas, garaje y trasteros. http://hdl.handle.net/10251/106611TFG

Posibilidades de la aplicación de la Arqueología de la Arquitectura en Santa Eulalia de Bóveda (Lugo)

This study attempts to show the possibilities offered by the application of Archaeology of Architecture in the Lugo building of Santa Eulalia de Bóveda. The present synthesis shows what we have identified as different architectonic stages which the building has seen since its construction. Each element has been isolated, whether decorative or architectonic elements present in the modern day construction. These are organized into five possible construction stages according to what has happened to the building from the Roman era to contemporary alterations.El trabajo que presentamos pretende mostrar las posibilidades que ofrece la aplicación de la Arqueología de la Arquitectura en el edificio lucense de Santa Eulalia de Bóveda. La presente síntesis muestra lo que hemos identificado como las diferentes etapas arquitectónicas por las que ha pasado el edificio desde su construcción. Se ha aislado cada uno de los elementos, tanto decorativos como arquitectónicos presentes en la actual construcción, organizándolos en las cinco posibles etapas constructivas por las que ha pasado el edificio, desde la etapa romana hasta la reforma contemporánea