Nou avenç en teràpia gènica

Un grup d'investigadors del CBATEG de la UAB, dirigit pel doctor Miguel Chillón, ha descrit el desenvolupament d'un sistema nou de producció d'adenovirus d'última generació (utilitzats com a vectors per a tractaments de teràpia gènica) que permet reduir, significativament, els inconvenients que existeixen fins ara per realitzar assaigs en humans. La UAB ha patentat recentment aquesta tècnica.Un grupo de investigadores del CBATEG de la UAB, dirigido por el doctor Miguel Chillón, ha descrito el desarrollo de un nuevo sistema de producción de adenovirus de última generación (utilizados como vectores para tratamientos de terapia génica) que permite reducir significativamente los inconvenientes existentes hasta ahora para realizar ensayos en humanos. La UAB ha patentado recientemente esta técnica.A group of CTABEG researchers, directed by Dr Miguel Chillon, describe the development of a new method for producing last-generation adenoviruses (used as vectors in gene therapy) which significantly reduce the inconveniences that existed until now in clinical trials carried out with humans. This method has been recently patented by the UAB

Functional characterization of the human mariner transposon Hsmar2

DNA transposons are mobile elements with the ability to mobilize and transport genetic information between different chromosomal loci. Unfortunately, most transposons copies are currently inactivated, little is known about mariner elements in humans despite their role in the evolution of the human genome, even though the Hsmar2 transposon is associated to hotspots for homologous recombination involved in human genetic disorders as Charcot-Marie-Tooth, Prader-Willi/Angelman, and Williams syndromes. This manuscript describes the functional characterization of the human HSMAR2 transposase generated from fossil sequences and shows that the native HSMAR2 is active in human cells, but also in bacteria, with an efficiency similar to other mariner elements. We observe that the sub-cellular localization of HSMAR2 is dependent on the host cell type, and is cytotoxic when overexpressed in HeLa cells. Finally, we also demonstrate that the binding of HSMAR2 to its own ITRs is specific, and that the excision reaction leaves non-canonical footprints both in bacteria and eukaryotic cells

Diffusion of culture on the Internet: world map of the online platforms

La cultura, como activo definitorio e identitario de una sociedad, necesita transmitirse para mantenerse activa y difundirse para ser (re)conocida y valorada. A través de la localización y el análisis de las ‘plataformas’ de difusión cultural en Internet, este artículo presenta un mapa mundial de las más relevantes atendiendo a cuatro parámetros de análisis: la ubicación geográfica, que permite una aproximación a la visión cartográfica de las plataformas; el uso de la lengua como soporte de manifestación y como vehículo de difusión en ediciones multilingües la titularidad de la iniciativa como agente de dinamización cultural desde la gestión pública o desde entidades privadas; y la concepción de la cultura como negocio a través de la construcción de la plataforma como herramienta de difusión o como escenario de negocio.The culture as a defining and identitary asset of a society must be transmitted to stay active and spread to be (re)known and appreciated. Through location and analysis of the "platforms" of cultural diffusion on the Internet, this paper presents a global map of the most important basis of four parameters of analysis: the geographic location, which allows to approach the cartographic vision of platforms, the use of language as support for expression and a vehicle for diffusion in multilingual editions the ownership of the initiative as an agent of cultural revitalization from public administration or from private entities; and the concept of culture as a business through the construction of the platform as a tool for diffusion or business scenario

Adeno-associated Virus Enhances Wild-type And Oncolytic Adenovirus Spread

The contamination of adenovirus (Ad) stocks with adeno-associated viruses (AAV) is usually unnoticed, and it has been associated with lower Ad yields upon large-scale production. During Ad propagation, AAV contamination needs to be detected routinely by polymerase chain reaction without symptomatic suspicion. In this study, we describe that the coinfection of either Ad wild type 5 or oncolytic Ad with AAV results in a large-plaque phenotype associated with an accelerated release of Ad from coinfected cells. This accelerated release was accompanied with the expected decrease in Ad yields in two out of three cell lines tested. Despite this lower Ad yield, coinfection with AAV accelerated cell death and enhanced the cytotoxicity mediated by Ad propagation. Intratumoral coinjection of Ad and AAV in two xenograft tumor models improved antitumor activity and mouse survival. Therefore, we conclude that accidental or intentional AAV coinfection has important implications for Ad-mediated virotherapy

PolyPurine Reverse Hoogsteen Hairpins Work as RNA Species for Gene Silencing

PolyPurine Reverse Hoogsteen Hairpins (PPRHs) are gene-silencing DNA-oligonucleotides developed in our laboratory that are formed by two antiparallel polypurine mirror repeat domains bound intramolecularly by Hoogsteen bonds. The aim of this work was to explore the feasibility of using viral vectors to deliver PPRHs as a gene therapy tool. After treatment with synthetic RNA, plasmid transfection, or viral infection targeting the survivin gene, viability was determined by the MTT assay, mRNA was determined by RT-qPCR, and protein levels were determined by Western blot. We showed that the RNA-PPRH induced a decrease in cell viability in a dose-dependent manner and an increase in apoptosis in PC-3 and HeLa cells. Both synthetic RNA-PPRH and RNA-PPRH intracellularly generated upon the transfection of a plasmid vector were able to reduce survivin mRNA and protein levels in PC-3 cells. An adenovirus type-5 vector encoding the PPRH against survivin was also able to decrease survivin mRNA and protein levels, leading to a reduction in HeLa cell viability. In this work, we demonstrated that PPRHs can also work as RNA species, either chemically synthesized, transcribed from a plasmid construct, or transcribed from viral vectors. Therefore, all these results are the proof of principle that viral vectors could be considered as a delivery system for PPRHs

bservation of the Physical Education teacher: a health classroom research

El objetivo es diseñar, validar y demostrar la aplicabilidad y eficacia de un instrumento observacional sobre la información transmitida por el profesor en el marco educativo del aula de Educación Física. Se diseñó el instrumento “Hoja de observación de vídeo: intervención didáctica” y se aplicó a la observación de cinco sesiones grabadas de Educación Física orientadas a contenidos de salud. El instrumento demostró validez y fiabilidad para analizar la información emitida por el profesor, que contenía contenidos referentes a salud y a otro tipo de contenidos. Se aporta un instrumento válido y fiable para analizar la información del profesorado en contenidos de salud y, además, se corrobora su aplicabilidad en el aula de Educación Física para la evaluación del profesor.The aim is to design, valid and show both feasibility and efficacy of an observational instrument on the information provided by the teacher in the Physical Education classroom. The instrument “Hoja de observación de vídeo: intervención didáctica” was designed and five recorded Physical Education lessons oriented to health content were observed. The instrument showed validity and reliability about the information provided by the teacher which compiled content related to health and other content. A valid and reliable instrument for analyzing the information from the teacher related to health content is provided and moreover, the instrument´s feasibility into the Physical Education classroom is confirmed for teacher evaluation.La autora Chillón Garzón disfrutó de una licencia por estudios financiada por la Consejería de Educación de la Junta de Andalucía

El gènere com a factor de variabilitat en les actituds envers la pràctica d’activitat físicoesportiva

L’orientació de les actituds que els adolescents manifesten envers l’activitat físicoesportiva representa un aspecte de gran importància a l’hora de promocionar-ne la pràctica com a hàbit de vida saludable, funció fonamental en la tasca educativa i, en concret, en l’Educació Física. Hem estudiat l’orientació de les actituds envers la pràctica físicoesportiva en 700 adolescents granadins d’entre 13 i 18 anys, en funció  del sexe, a partir del “qüestionari CAAFS”. Els resultats mostren que els nois tenen nivells actitudinals significativament superiors (p < 0.05) als de les noies, en les actituds orientades tant cap al procés (3,69 i 3,45, respectivament) com cap al resultat (2,45 ells i 2,33 elles). La “millora de l’aparença” (2,71) és la dimensió de l’actitud cap a la que les adolescents granadines manifesten més tendència, i la dimensió “victòria” ho és en el cas dels barons (2,65). Per tant, es pot concloure indicant la necessitat de millorar les actituds envers la pràctica físicoesportiva en les noies i considerar la necessitat de diversificar la pràctica d’acord amb la seva orientació en funció del gènere

El género como factor de variabilidad en las actitudes hacia la práctica de actividad físico-deportiva

La orientación de las actitudes que los adolescentes manifiestan hacia la actividad físico-deportiva representa un aspecto de gran importancia a la hora de promocionar su práctica como hábito de vida saludable, función fundamental en la labor educativa y, en concreto, en la Educación Física. En 700 adolescentes granadinos de entre 13 y 18 años, se ha estudiado la orientación de las actitudes hacia la práctica físico-deportiva en función del sexo a partir del “cuestionario CAAFS”. Los resultados muestran que los chicos tienen niveles actitudinales significativamente superiores (p < 0.05) a los de las chicas en las actitudes orientadas tanto hacia el proceso (3,69 y 3,45, respectivamente) como hacia el resultado (2,45 ellos y 2,33 ellas). La “mejora de la apariencia” (2,71) es la dimensión actitudinal hacia la que las adolescentes granadinas manifiestan una mayor tendencia, y la dimensión “victoria” lo es en el caso de los varones (2,65). Por tanto se puede concluir indicando la necesidad de mejorar las actitudes hacia la práctica físico-deportiva en las chicas y considerar la necesidad de diversificar la práctica en función de la orientación de las mismas en función del género

Cerebellar astrocyte transduction as gene therapy for megalencephalic leukoencephalopathy

Megalencephalic leukoencephalopathy with subcortical cysts (MLC) is a rare genetic disorder belonging to the group of vacuolating leukodystrophies. It is characterized by megalencephaly, loss of motor functions, epilepsy, and mild mental decline. In brain biopsies of MLC patients, vacuoles were observed in myelin and in astrocytes surrounding blood vessels. There is no therapy for MLC patients, only supportive treatment. We show here a preclinical gene therapy approach for MLC using the Mlc1 knock-out mouse. An adeno-associated virus coding for human MLC1 under the control of the glial fibrillary acidic protein promoter was injected in the cerebellar subarachnoid space of Mlc1 knock-out and wild-type animals at 2 months of age, before the onset of the disease, as a preventive approach. We also tested a therapeutic strategy by injecting the animals at 5 months, once the histopathological abnormalities are starting, or at 15 months, when they have progressed to a more severe pathology. MLC1 expression in the cerebellum restored the adhesion molecule GlialCAM and the chloride channel ClC-2 localization in Bergmann glia, which both are mislocalized in Mlc1 knock-out model. More importantly, myelin vacuolation was extremely reduced in treated mice at all ages and correlated with the amount of expressed MLC1 in Bergmann glia, indicating not only the preventive potential of this strategy but also its therapeutic capacity. In summary, here we provide the first therapeutic approach for patients affected with MLC. This work may have also implications to treat other diseases affecting motor function such as ataxias

Differential toxicity profile of secreted and processed α-Klotho expression over mineral metabolism and bone microstructure

The aging-protective gene α-Klotho (KL) produces two main transcripts. The full-length mRNA generates a transmembrane protein that after proteolytic ectodomain shedding can be detected in serum as processed Klotho (p-KL), and a shorter transcript which codes for a putatively secreted protein (s-KL). Both isoforms exhibit potent pleiotropic beneficial properties, although previous reports showed negative side effects on mineral homeostasis after increasing p-KL concentration exogenously. Here, we expressed independently both isoforms using gene transfer vectors, to assess s-KL effects on mineral metabolism. While mice treated with p-KL presented altered expression of several kidney ion channels, as well as altered levels of Pi and Ca2+ in blood, s-KL treated mice had levels comparable to Null-treated control mice. Besides, bone gene expression of Fgf23 showed a fourfold increase after p-KL treatment, effects not observed with the s-KL isoform. Similarly, bone microstructure parameters of p-KL-treated mice were significantly worse than in control animals, while this was not observed for s-KL, which showed an unexpected increase in trabecular thickness and cortical mineral density. As a conclusion, s-KL (but not p-KL) is a safe therapeutic strategy to exploit KL anti-aging protective effects, presenting no apparent negative effects over mineral metabolism and bone microstructure