El ozono no consigue la desinfección de los vehículos de emergencias de virus similares al SARS-CoV-2 / Ozone fails to disinfect emergency vehicles contaminated with viruses similar to SARS-CoV-2

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La inflamación inhibe la expresión vascular de la fibulina-5: implicación del factor de transcripción SOX9 / Inflammation inhibits vascular fibulin-5 expression: Involvement of transcription factor SOX9

Introducción: La fibulina-5 (FBLN5) es una proteína elastogénica implicada en el remodelado de la matriz extracelular (MEX), un proceso fundamental en el aneurisma de aorta abdominal (AAA). Sin embargo, no se ha determinado la posible contribución de la FBLN5 al AAA. Métodos: Se realizaron análisis por PCR a tiempo real, Western blot, transducción lentiviral, transfección transitoria e inmunoprecipitación de cromatina (ChIP) en aorta abdominal de pacientes con AAA o donantes y en células musculares lisas de aorta humana (CMLV). Resultados: La expresión vascular de la FBLN5 disminuye en la aorta abdominal de pacientes con AAA frente a donantes sanos. El nivel de ARNm y proteína de la FBLN5 y su secreción al espacio extracelular se redujeron en CMLV expuestas a estímulos inflamatorios. Este efecto se produce a través de un mecanismo transcripcional en el que está implicada una región proximal del promotor de la FBLN5 que contiene un elemento de respuesta a SOX. De hecho, la expresión de SOX9 se inhibe en CMLV tratadas con LPS y TNFα y disminuye en el AAA, en el que correlaciona con la de la FBLN5. Además, la sobreexpresión de SOX9 contrarrestó la disminución de la expresión y actividad transcripcional de la FBLN5 inducida por el TNFα. Finalmente, observamos que SOX9 interacciona con el promotor de la FBLN5 y que esta unión se reduce en respuesta a TNFα. Conclusiones: La inhibición de la FBLN5 en el AAA humano podría contribuir al remodelado destructivo de la matriz extracelular inducido por el componente inflamatorio de la patología

Introduction: Fibulin-5 (FBLN5) is an elastogenic protein critically involved in extracellular matrix (ECM) remodelling, a key process in abdominal aortic aneurysm (AAA). However, the possible contribution of FBLN5 to AAA development has not been addressed. Methods: Expression levels were determined by real-time PCR and Western blot in human abdominal aorta from patients with AAA or healthy donors, as well as in human aortic vascular smooth muscle cells (VSMC). Lentiviral transduction, transient transfections, and chromatin immunoprecipitation (ChIP) assays were also performed. Results: The expression of FBLN5 in human AAA was significantly lower than in healthy donors. FBLN5 mRNA and protein levels and their secretion to the extracellular environment were down-regulated in VSMC exposed to inflammatory stimuli. Interestingly, FBLN5 transcriptional activity was inhibited by TNFα and lipopolysaccharide (LPS), and depends on a SOX response element. In fact, SOX9 expression was reduced in VMSC induced by inflammatory mediators and in human AAA, and correlated with that of FBLN5. Furthermore, SOX9 over-expression limited the reduction of FBLN5 expression induced by cytokines in VSMC. Finally, it was observed that SOX9 interacts with FBLN5 promoter, and that this binding was reduced upon TNFα exposure. Conclusions: FBLN5 downregulation in human AAA could contribute to extracellular matrix remodelling induced by the inflammatory component of the disease

Generación de células mesenquimales con potencial osteogénico mejorado genéticamente para el tratamiento de enfermedades que cursan con defectos de masa ósea / Generation of human mesenchymal cells with genetically improved osteogenic potential for treatment of bone diseases

Objetivo: Generar células madre mesenquimales humanas (MSCs) modificadas para optimizar su potencial de diferenciación osteogénica, destinadas a su empleo en implantes cerámicos para regeneración ósea. Material y método: Se emplearon ratones inmunodeficientes NOD/SCID (3-6 ratones por condición experimental y ensayo) y se generaron vectores lentivirales basados en la recombinasa Cre, que sobreexpresan el factor regulador del proceso osteogénico Dlx5 (o la proteína fluorescente GFP como control) de forma autolimitada en el tiempo. Estos vectores se utilizaron para transducir hMSCs, y su potencial osteogénico se analizó in vitro e in vivo en un modelo de formación de hueso heterotópico en ratón. Resultados: Las hMSCs transducidas con los vectores que expresan Dlx5 de forma autolimitada fueron capaces de diferenciarse eficientemente a hueso de forma espontánea, de manera similar a las hMSCs control en presencia del factor osteoinductor BMP-2. Conclusión: Hemos desarrollado un sistema de modificación de hMSCs para aumentar su potencial osteogénico que consiste en un vector lentiviral que expresa el factor osteoinductor Dlx5 de forma autolimitada. Las hMSCs modificadas diferencian a hueso de manera eficiente, tanto in vitro como in vivo (AU)

Objective: This article proposes the generation of modified human mesenchymal stem cells (hMSCs) for optimizing their osteogenic differentiation potential, in order to be employed in ceramic implants for bone regeneration. Material and method: We have generated lentiviral vectors based on Cre recombinase, which lead to overexpression of a regulatory factor of osteogenic process, Dlx5 (or GFP fluorescent protein as a control), in a selflimited fashion. We have transduced hMSCs with these vectors, and we have analyzed their osteogenic potential both in vitro and in vivo in a model of heterotopic bone formation in mice. For this purpose we have used immunodeficient NOD/SCID mice (3-6 mice per condition and experiment). Results: hMSCs transduced with self-limited Dlx5-expressing vectors efficiently differentiate into bone in vitro and in vivo, similar to control hMSCs in the presence of osteoinductive factor BMP-2. Conclusion: We have developed a system to modify hMSCs in order to improve their osteogenic potential. This system consists of a lentiviral vector which expresses osteoinductive factor Dlx5 in a self-limited fashion. Modified hMSCs efficiently differentiate into bone, both in vitro and in vivo (AU)

Downregulation of KLF8 expression by shRNA induces inhibition of cell proliferation in CAL27 human oral cancer cells

Objectives KLF8 is a member of KLF transcription factors which play an important tolr in oncogenesis. It is barely expressed in normal human epithelial cells but highly overexpressed in several types of human cancer cell lines. In the present study, we investigate the role of KLF8 in oral cancer and the effects of KLF8 knockdown via lentivirus mediated siRNA infection in human adenosquamos carcinoma CAL 27 cells. Study Design We developed a vector-based siRNA expression system that can induce RNAi in CAL 27 oral cancer cells. Downregulation of KLF8 was confirmed by evaluating GFP expressions, RT-PCR and western blot analysis. Finally, the effects of KLF8 downregulation were analyzed by MTT assay and colony formation assays. Results The expression levels of KLF8 mRNA and proteins are reduced in CAL 27 cells that transfected with 21-nt siRNA against KLF8. Lentivirus-mediated silencing of KLF8 reduces cell proliferation and colonies number, thereby indicating the role of KLF8 in cell proliferation and tumorigenesis. Conclusions These results strongly suggest that KLF8 is essential for growth of CAL 27 cancer cells. A better understanding of KLF8 function and processing may provide novel insights into the clinical therapy of oral cancer (AU)

Terapia génica en la enfermedad de Parkinson / Gene therapy for Parkinson's disease

La enfermedad de Parkinson es un trastorno crónico y progresivo cuyo tratamiento no impide a medio plazo la aparición de complicaciones motoras y psíquicas invalidantes. Las técnicas de terapia génica de aplicación cada vez mayor en el campo de las enfermedades neurodegenerativas se suman a las posibilidades de tratamiento de esta patología. Entre las modalidades existentes, las estrategias in vivo que emplean potentes vectores virales son las que mejores resultados han obtenido en los distintos modelos existentes de la enfermedad. Este artículo pretende revisar la información referente al empleo de estas últimas técnicas, los ensayos terapéuticos que se han llevado a cabo y las ventajas e inconvenientes que tiene la utilización de los diferentes vectores

Parkinson's disease is a chronic and progressive disorder whose treatment does not prevent middle term appearance of invalidating motor and psychic complications. Gene therapy techniques which are increasingly applied in the field of neurodegenerative diseases are added to the possibility of treatment of this disease. Among the existing modalities, the in vivo strategies that use potent viral vectors are those which have obtained the best results in the different existing models of the disease. This article aims to review the information regarding the use of these latter techniques, the therapeutic trials that have been conducted and the advantages and disadvantages that the use of the different vectors have

Terapia génica de la hemofilia: recapitulación en los albores de un nuevo milenio / Gene therapy in hemophilia. Summary of the dawning of a new millenium

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