Desarrollo de un Laboratorio Virtual para la realización de e-comparisons en los ensayos de aislamiento acústico de fachadas según UNE-EN ISO 140-5

Actualmente son una práctica común los procesos de normalización de métodos de ensayo y acreditación de laboratorios, ya que permiten una evaluación de los procedimientos llevados a cabo por profesionales de un sector tecnológico y además permiten asegurar unos mínimos de calidad en los resultados finales. En el caso de los laboratorios de acústica, para conseguir y mantener la acreditación de un laboratorio es necesario participar activamente en ejercicios de intercomparación, utilizados para asegurar la calidad de los métodos empleados. El inconveniente de estos ensayos es el gran coste que suponen para los laboratorios, siendo en ocasiones inasumible por estos teniendo que renunciar a la acreditación. Este Proyecto Fin de Grado se centrará en el desarrollo de un Laboratorio Virtual implementado mediante una herramienta software que servirá para realizar ejercicios de intercomparación no presenciales, ampliando de ese modo el concepto e-comparison y abriendo las bases a que en un futuro este tipo de ejercicios no presenciales puedan llegar a sustituir a los llevados a cabo actualmente. En el informe primero se hará una pequeña introducción, donde se expondrá la evolución y la importancia de los procedimientos de calidad acústica en la sociedad actual. A continuación se comentará las normativas internacionales en las que se soportará el proyecto, la norma ISO 145-5, así como los métodos matemáticos utilizados en su implementación, los métodos estadísticos de propagación de incertidumbres especificados por la JCGM (Joint Committee for Guides in Metrology). Después, se hablará sobre la estructura del proyecto, tanto del tipo de programación utilizada en su desarrollo como la metodología de cálculo utilizada para conseguir que todas las funcionalidades requeridas en este tipo de ensayo estén correctamente implementadas. Posteriormente se llevará a cabo una validación estadística basada en la comparación de unos datos generados por el programa, procesados utilizando la simulación de Montecarlo, y unos cálculos analíticos, que permita comprobar que el programa funciona tal y como se ha previsto en la fase de estudio teórico. También se realizará una prueba del programa, similar a la que efectuaría un técnico de laboratorio, en la que se evaluará la incertidumbre de la medida calculándola mediante el método tradicional, pudiendo comparar los datos obtenidos con los que deberían obtenerse. Por último, se comentarán las conclusiones obtenidas con el desarrollo y pruebas del Laboratorio Virtual, y se propondrán nuevas líneas de investigación futuras relacionadas con el concepto e-comparison y la implementación de mejoras al Laboratorio Virtual. ABSTRACT. Nowadays it is common practise to make procedures to normalise trials methods standards and laboratory accreditations, as they allow for the evaluation of the procedures made by professionals from a particular technological sector in addition to ensuring a minimum quality in the results. In order for an acoustics laboratory to achieve and maintain the accreditation it is necessary to actively participate in the intercomparison exercises, since these are used to assure the quality of the methods used by the technicians. Unfortunately, the high cost of these trials is unaffordable for many laboratories, which then have to renounce to having the accreditation. This Final Project is focused on the development of a Virtual Laboratory implemented by a software tool that it will be used for making non-attendance intercomparison trials, widening the concept of e-comparison and opening the possibility for using this type of non-attendance trials instead of the current ones. First, as a short introduction, I show the evolution and the importance today of acoustic quality procedures. Second, I will discuss the international standards, such as ISO 145-5, as well the mathematic and statistical methods of uncertainty propagation specified by the Joint Committee for Guides in Metrology, that are used in the Project. Third, I speak about the structure of the Project, as well as the programming language structure and the methodology used to get the different features needed in this acoustic trial. Later, a statistical validation will be carried out, based on comparison of data generated by the program, processed using a Montecarlo simulation, and analytical calculations to verify that the program works as planned in the theoretical study. There will also be a test of the program, similar to one that a laboratory technician would carry out, by which the uncertainty in the measurement will be compared to a traditional calculation method so as to compare the results. Finally, the conclusions obtained with the development and testing of the Virtual Laboratory will be discussed, new research paths related to e-comparison definition and the improvements for the Laboratory will be proposed.

La Traca : semanari pa la gent de tro: Año IX Número 140 - 3 enero 1892

Recurso electrónico. Valencia: BVNP, 2014

The Mr 140,000 Intermediate Chain of Chlamydomonas Flagellar Inner Arm Dynein Is a WD-Repeat Protein Implicated in Dynein Arm Anchoring

Previous structural and biochemical studies have revealed that the inner arm dynein I1 is targeted and anchored to a unique site located proximal to the first radial spoke in each 96-nm axoneme repeat on flagellar doublet microtubules. To determine whether intermediate chains mediate the positioning and docking of dynein complexes, we cloned and characterized the 140-kDa intermediate chain (IC140) of the I1 complex. Sequence and secondary structural analysis, with particular emphasis on β-sheet organization, predicted that IC140 contains seven WD repeats. Reexamination of other members of the dynein intermediate chain family of WD proteins indicated that these polypeptides also bear seven WD/β-sheet repeats arranged in the same pattern along each intermediate chain protein. A polyclonal antibody was raised against a 53-kDa fusion protein derived from the C-terminal third of IC140. The antibody is highly specific for IC140 and does not bind to other dynein intermediate chains or proteins in Chlamydomonas flagella. Immunofluorescent microscopy of Chlamydomonas cells confirmed that IC140 is distributed along the length of both flagellar axonemes. In vitro reconstitution experiments demonstrated that the 53-kDa C-terminal fusion protein binds specifically to axonemes lacking the I1 complex. Chemical cross-linking indicated that IC140 is closely associated with a second intermediate chain in the I1 complex. These data suggest that IC140 contains domains responsible for the assembly and docking of the I1 complex to the doublet microtubule cargo.

The Chlamydomonas IDA7 Locus Encodes a 140-kDa Dynein Intermediate Chain Required to Assemble the I1 Inner Arm Complex

To identify new loci that are involved in the assembly and targeting of dynein complexes, we have screened a collection of motility mutants that were generated by insertional mutagenesis. One such mutant, 5B10, lacks the inner arm isoform known as the I1 complex. This isoform is located proximal to the first radial spoke in each 96-nm axoneme repeat and is an important target for the regulation of flagellar motility. Complementation tests reveal that 5B10 represents a new I1 locus, IDA7. Biochemical analyses confirm that ida7 axonemes lack at least five I1 complex subunits. Southern blots probed with a clone containing the gene encoding the 140-kDa intermediate chain (IC) indicate that the ida7 mutation is the result of plasmid insertion into the IC140 gene. Transformation with a wild-type copy of the IC140 gene completely rescues the mutant defects. Surprisingly, transformation with a construct of the IC140 gene lacking the first four exons of the coding sequence also rescues the mutant phenotype. These studies indicate that IC140 is essential for assembly of the I1 complex, but unlike other dynein ICs, the N-terminal region is not critical for its activity.

A truncated mutant (residues 58-140) of the hepatitis B virus X protein retains transactivation function.

The hepatitis B virus X protein (HBx) sequence (154 aa) has been divided into six regions (A-F) based on its sequence homology with X proteins of other mammalian hepadnaviruses. Regions A, C, and E are more conserved and include all the four conserved cysteines (C7, C61, C69, and C137). To localize the regions of HBx important for transactivation, a panel of 10 deletion mutants (X5-X14) and 4 single point mutants (X1-X4), each corresponding to a conserved cysteine residue, was constructed by site-directed mutagenesis. A HBx-specific monoclonal antibody was developed and used to confirm the expression of mutants by Western blot. Transactivation property of the HBx mutants was studied on Rous sarcoma virus-long terminal repeat (RSV-LTR) in transient transfection assays. We observed that deletion of the most conserved region A or substitution of the N-terminal cysteine (C7) had no effect on transactivation. Deletion of the nonconserved regions B or F also had no deleterious effects. Deletions of regions C and D resulted in a significant loss of function. Substitution of both C61 and C69 present in region C, caused almost 90% loss of activity that could be partially overcome by transfecting more expression plasmid. The fully conserved 9 amino acid segment (residues 132 to 140) within region E including C137 appeared to be crucial for its activity. Finally, a truncated mutant X15 incorporating only regions C to E (amino acids 58-140) was able to stimulate the RSV-LTR quite efficiently, suggesting a crucial role played by this domain in transactivation function.