6 resultados para Quantitative Real-time Pcr

em Archivo Digital para la Docencia y la Investigación - Repositorio Institucional de la Universidad del País Vasco


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Background: Staphyloccocal nuclease domain-containing protein 1 (SND1) is involved in the regulation of gene expression and RNA protection. While numerous studies have established that SND1 protein expression is modulated by cellular stresses associated with tumor growth, hypoxia, inflammation, heat- shock and oxidative conditions, little is known about the factors responsible for SND1 expression. Here, we have approached this question by analyzing the transcriptional response of human SND1 gene to pharmacological endoplasmic reticulum (ER) stress in liver cancer cells. Results: We provide first evidence that SND1 promoter activity is increased in human liver cancer cells upon exposure to thapsigargin or tunicamycin or by ectopic expression of ATF6, a crucial transcription factor in the unfolded protein response triggered by ER stress. Deletion analysis of the 5'-flanking region of SND1 promoter identified maximal activation in fragment (-934, +221), which contains most of the predicted ER stress response elements in proximal promoter. Quantitative real- time PCR revealed a near 3 fold increase in SND1 mRNA expression by either of the stress- inducers; whereas SND1 protein was maximally upregulated (3.4-fold) in cells exposed to tunicamycin, a protein glycosylation inhibitor. Conclusion: Promoter activity of the cell growth- and RNA-protection associated SND1 gene is up-regulated by ER stress in human hepatoma cells.

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The presence of endogenous opioid peptides in different testicular cell types has been extensively characterized and provides evidence for the participation of the opioid system in the regulation of testicular function. However, the exact role of the opioid system during the spermatogenesis has remained controversial since the presence of the mu-, delta-and kappa-opioid receptors in spermatogenic cells was yet to be demonstrated. Through a combination of quantitative real-time PCR, immunofluorescence, immunohistochemistry and flow cytometry approaches, we report for the first time the presence of active mu-, deltaand kappa-opioid receptors in mouse male germ cells. They show an exposition time-dependent response to opioid agonist, hence suggesting their active involvement in spermatogenesis. Our results contribute to understanding the role of the opioid receptors in the spermatogenesis and could help to develop new strategies to employ the opioid system as a biochemical tool for the diagnosis and treatment of male infertility.

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A sliding mode position control for high-performance real-time applications of induction motors in developed in this work. The design also incorporates a simple flux estimator in order to avoid the flux sensors. Then, the proposed control scheme presents a low computational cost and therefore can be implemented easily in a real-time applications using a low cost DSP-processor. The stability analysis of the controller under parameter uncertainties and load disturbances in provided using Lyapunov stability theory. Finally, simulated and experimental results show that the proposed controller with the proposed observer provides a good trajectory tracking and that this scheme is robust with respect to plant parameter variations and external load disturbances.

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A lo largo de este documento, se va a explicar la implantación del proyecto que he realizado basado en la localización de vehículos en la fábrica de Mercedes Benz España situada en Vitoria-Gasteiz. Durante la realización de este proyecto, se han llevado a cabo diversos estudios con el fin de conseguir la correcta implantación de las tecnologías empleadas. Se han realizado diferentes alternativas de posicionamiento de los componentes y diversas pruebas para comprobar el correcto funcionamiento de la solución. La solución del proyecto se realizará en distintas fases. La primera de ellas tratará sobre el estudio en una determinada zona de la fábrica, más concretamente la denominada “Área Técnica”, en esta zona se encuentran los vehículos que sufren algún retoque una vez están montados, esta zona se utilizará como piloto para una vez finalizado y comprobado su éxito ampliar la solución al resto de zonas. Previamente a mi incorporación se realizó un estudio para la colocación de los elementos necesarios en esta zona y se ha visto las posibilidades y beneficios que aportaría el control de los vehículos dentro de la fábrica. La siguiente fase será implantar la solución en el resto de las áreas que se encuentran dentro de la fábrica de Vitoria-Gasteiz así como la instalación de unos dispositivos que estarán ubicados en las puertas. Estos ayudarán a mejorar la ubicación de los vehículos ya que podremos conocer si los vehículos se encuentran dentro o fuera de la fábrica. Finalmente se ha realizado la integración de la solución en los sistemas actuales que utilizan en la fábrica para la gestión de los vehículos durante su ciclo de vida.

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In this paper, a real time sliding mode control scheme for a variable speed wind turbine that incorporates a doubly feed induction generator is described. In this design, the so-called vector control theory is applied, in order to simplify the system electrical equations. The proposed control scheme involves a low computational cost and therefore can be implemented in real-time applications using a low cost Digital Signal Processor (DSP). The stability analysis of the proposed sliding mode controller under disturbances and parameter uncertainties is provided using the Lyapunov stability theory. A new experimental platform has been designed and constructed in order to analyze the real-time performance of the proposed controller in a real system. Finally, the experimental validation carried out in the experimental platform shows; on the one hand that the proposed controller provides high-performance dynamic characteristics, and on the other hand that this scheme is robust with respect to the uncertainties that usually appear in the real systems.

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The potential of the 18S rRNA V9 metabarcoding approach for diet assessment was explored using MiSeq paired-end (PE; 2 9 150 bp) technology. To critically evaluate the method's performance with degraded/digested DNA, the diets of two zooplanktivorous fish species from the Bay of Biscay, European sardine (Sardina pilchardus) and European sprat (Sprattus sprattus), were analysed. The taxonomic resolution and quantitative potential of the 18S V9 metabarcoding was first assessed both in silico and with mock and field plankton samples. Our method was capable of discriminating species within the reference database in a reliable way providing there was at least one variable position in the 18S V9 region. Furthermore, it successfully discriminated diet between both fish species, including habitat and diel differences among sardines, overcoming some of the limitations of traditional visual-based diet analysis methods. The high sensitivity and semi-quantitative nature of the 18S V9 metabarcoding approach was supported by both visual microscopy and qPCR-based results. This molecular approach provides an alternative cost and time effective tool for food-web analysis.