Engineered Salmonella allows real-time heterologous gene expression monitoring within infected zebrafish embryos

Short communication.Microbial host–pathogen interactions have been traditionally well studied at genetic and physiological levels, but cell-resolution analyses have been particularly scarce. This has been especially remarkable for intracellular parasites for two major reasons: first, the inherent loss of bacteria traceability once infects its hosts; second and more important, the limited availability of genetic tools that allow a tight regulated expression of bacterial virulence genes once inside the host tissues. Here we present novel data supporting the use of zebrafish embryos to monitor Salmonella enterica serovar Thyphimurium infection. Intravenous infection of Salmonella can be easily monitored using in vivo fluorescence that allows the visualization of free-swimming bacteria through the circulatory system. Moreover, we have engineered Salmonella to voluntarily activate heterologous gene expression at any point during infection once inside the zebrafish macrophages using a salicylate-based expression system. This approach allows real-time cell-resolution in vivo monitoring of the infection. All together, this approach paves the road to cell-based resolution experiments that would be harder to mimic in other vertebrate infection models.his work was funded by grants BFU2010-14839, CSD2007-00008, CSD2007-00005 and Proyectos de Excelencia CVI-3488 and P07-CVI-02518 from the Spanish and Andalusian Governments, respectively. Jose Luis Royo holds a JAE DOC contract from the Spanish National Research Council (CSIC).Peer reviewe

Identification and Analysis of Conserved cis-Regulatory Regions of the MEIS1 Gene

Meis1, a conserved transcription factor of the TALE-homeodomain class, is expressed in a wide variety of tissues during development. Its complex expression pattern is likely to be controlled by an equally complex regulatory landscape. Here we have scanned the Meis1 locus for regulatory elements and found 13 non-coding regions, highly conserved between humans and teleost fishes, that have enhancer activity in stable transgenic zebrafish lines. All these regions are syntenic in most vertebrates. The composite expression of all these enhancer elements recapitulate most of Meis1 expression during early embryogenesis, indicating they comprise a basic set of regulatory elements of the Meis1 gene. Using bioinformatic tools, we identify a number of potential binding sites for transcription factors that are compatible with the regulation of these enhancers. Specifically, HHc2:066650, which is expressed in the developing retina and optic tectum, harbors several predicted Pax6 sites. Biochemical, functional and transgenic assays indicate that pax6 genes directly regulate HHc2:066650 activity.This work was funded through grants BFU2009-07044 (MICINN) and Proyecto de Excelencia CVI 2658 (Junta de Andalucía) to FC and BFU2010-14839 (MICINN), CSD2007-00008 and Proyecto de Excelencia CVI-3488 to JLGS. JLR is a recipient of a JAE-DOC contract from the Spanish National Research Council (CSIC)

Los derechos on-line en el ordenamiento laboral español : estado de la cuestión

Los derechos on-line son una categoría jurídica de reciente creación, que incluye diversos derechos relacionados con el uso de TIC en la empresa. Entre éstos pueden encontrarse tanto derechos de acceso como de privacidad. Una de las prioridades de los sindicatos europeos es el reconocimiento y la protección legal de estos derechos, para lo que han tomado una serie de iniciativas de distinto tipo. La situación de estos derechos en el ordenamiento laboral español es muy deficientes, y ni el Derecho positivo ni la jurisprudencia laboral han sido capaces hasta ahora de darles un tratamiento válido. Sólo la negociación colectiva ha sido capaz de dar algunas respuestas a los problemas de acceso y privacidad, si bien de una manera limitada, y en un número pequeño de convenios.Online rights are a legal category of recent creation, that includes diverse rights related to the use of TIC in the company. Among these, rights of access and privacy can be found. One of the trade unions priorities is the recognition and legal protection of these rights, for which they have taken a series of different types of initiatives. These right’s situation in the Spanish legal order are quite poor, and neither the positive law nor the labour jurisprudence have been able to provide them with a valid treatment. Only the collective negotiation has been able to give some answers to the accessibility and privacy issues, although in a limited way and in a small number of agreements

Exploitation of prokaryotic expression systems based on the salicylate-dependent control circuit encompassing nahR/Psal ::xylS2 for biotechnological applications

This is an open-access article licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported License.Expression vectors appear to be an indispensable tool for both biological studies and biotechnological applications. Controlling gene overexpression becomes a critical issue when protein production is desired. In addition to several aspects regarding toxicity or plasmid instability, tight control of gene expression is an essential factor in biotechnological processes. Thus, the search for better-controlled circuits is an important issue among biotechnologists. Traditionally, expression systems involve a single regulatory protein operating over a target promoter. However, these circuits are limited on their induction ratios (e.g., by their restriction in the maximal expression capacity, by their leakiness under non-induced conditions). Due to these limitations, regulatory cascades, which are far more efficient, are necessary for biotechnological applications. Thus, regulatory circuits with two modules operating in cascade offer a significant advantage. In this review, we describe the regulatory cascade based on two salicylate-responsive transcriptional regulators of Pseudomonas putida (nahR/P sal::xylS2), its properties, and contribution to a tighter control over heterologous gene expression in different applications. Nowadays, heterologous expression has been proven to be an indispensable tool for tackling basic biological questions, as well as for developing biotechnological applications. As the nature of the protein of interest becomes more complex, biotechnologists find that a tight control of gene expression is a key factor which conditions the success of the downstream purification process, as well as the interpretation of the results in other type of studies. Fortunately, different expression systems can be found in the market, each of them with their own pros and cons. In this review we discuss the exploitation of prokaryotic expression systems based on a promising expression system, the salicylate-dependent control circuit encompassing nahR/P sal::xylS2, as well as some of the improvements that have been done on this system to exploit it more efficiently in the context of both biotechnological applications and basic research.Peer Reviewe

Evaluación de la capacidad de cubrición de los moruecos

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Relación entre la condición física cardiovascular y la distribución de grasa en niños y adolescentes

Fundamentos: El objetivo de este estudio es determinar la relación entre el grado de condición física cardiovascular y la acumulación y distribución de masa grasa en niños y niñas de 7 a 17 años. Material y métodos: La muestra se compone de 1.625 niños y 1.244 niñas, a los que se determinó la potencia aeróbica máxima mediante el test de carrera de la Course Navette. Previamente, se realizó una evaluación antropométrica con el fin de poder establecer la cantidad total de masa grasa subcutánea acumulada (suma de 6 pliegues corporales), así como la cantidad de masa grasa subcutánea acumulada en la región del tronco (suma de 3 pliegues del tronco). Se formaron 2 grupos en función de la condición física cardiovascular. Los sujetos que obtuvieron menores valores de VO2 máx (dos últimos quintiles) formaron parte del grupo de baja condición física cardiovascular, mientras que aquellos cuyos valores de VO2 máx se encontraron entre los dos quintiles superiores formaron parte del grupo de alta condición física cardiovascular. Resultados: Una vez ajustados por las diferencias de edad, talla y masa corporal, los resultados de este estudio demuestran que tanto en niños como en niñas un grado superior de condición física cardiovascular se asocia con cantidades significativamente menores de grasa subcutánea no sólo en el cuerpo entero, sino particularmente en la zona del tronco. Conclusiones: En los niños y niñas con una mejor condición física cardiovascular se observa una composición corporal más saludable y un menor riesgo de accidente cardiovascular. Es importante incorporar el grado de condición física en la valoración del riesgo de salud en las poblaciones estudiadas

Information and communication technologies for the education of people with cognitive disabilities

Este artículo resume los trabajos de investigación y las conclusiones obtenidas relacionadas con el uso de las Tecnologías de la Información y de la Comunicación (TIC) en personas con discapacidad cognitiva. Los resultados de los estudios realizados por nuestro grupo de investigación se centraron en la problemática de la usabilidad y accesibilidad dentro de la generación de material didáctico y formativo audiovisual interactivo, factores imprescindibles para un diseño formal y funcional óptimo de herramientas telemáticas, dirigidas a permitir que, en un principio, cualquier tipo de usuario pueda acceder a ellas, independientemente de si padecen una discapacidad o no. Estos estudios intentaban valorar, por un lado, el grado de satisfacción de un grupo de pacientes con una enfermedad mental con deterioro cognitivo de carácter crónico al enfrentarse al manejo de herramientas digitales multimedia para su formación educacional, laboral y terapéutica, y por otro, valorar el grado de usabilidad y accesibilidad empleados en aplicaciones informáticas para evaluar y analizar las limitaciones concretas de estos colectivos, con la finalidad de componer un corpus teórico con criterios de usabilidad y accesibilidad para personas con una enfermedad mental que complementen las pautas generales de diseño y programación utilizados actualmente. Si bien es cierto que existen numerosas investigaciones en el ámbito médico y sanitario sobre la adecuación de las TIC a las necesidades de colectivos con alguna discapacidad física, existe una gran carencia de estudios científicos que valoren y analicen la accesibilidad real en la utilización de las nuevas tecnologías en personas con algún tipo de discapacidad intelectual de carácter cognitivo, centrado exclusivamente, en personas con algún grado de enfermedad mental. La evaluación sistemática de la usabilidad permite a diseñadores y desarrolladores informáticos valorar y mejorar, de forma continuada, los diseños y desarrollos obtenidos a través de la experiencia con el usuario, para optimizar el diseño y conseguir, de esta manera, un producto más competitivo, especialmente en el caso del acceso a Internet y de la educación a distancia.This article summarizes the research and conclusions related to the use of Information and Communication Technology (ICT) for people with cognitive disabilities. The results of studies conducted by our research group focused on issues of usability and accessibility within the generation of audio-visual materials and interactive training the essential factors for optimal format and function design of telematic tools, in order to allow all users access initially, whether a disable or not. These studies set out to assess, firstly, the degree of satisfaction of a group of patients with mental disabilities to manage digital media tools for their education, work and therapy, and secondly, the degree of usability and accessibility in software used to evaluate and analyze the constraints of these groups, in order to compose a corpus theoretical criteria of usability and accessibility for people with a mental illness that compliments general design guidelines and programming used today. While there are numerous investigations swap round medical and health fields on the appropriateness of ITCs to meet the needs of groups with physical disabilities, there is a serious lack of scientific studies that evaluate and analyze the real accessibility of the use of new technologies for people with intellectual disabilities, focusing exclusively on people with some degree of mental illness. The systematic assessment of usability allows software designers and developers to continuously evaluate, improve and optimize the desing and development using the experience gained with the user to optimize the design and achieve, in this way, a competitive product is, therefore, achievable, especially in the case of Internet access and distance.peerReviewe