Fotogrametría de UAV de ala fija y comparación con topografía clásica.

El objetivo de este proyecto es realizar un estudio comparativo utilizando dos procedimientos diferentes para la obtención de mapas y modelos digitales del terreno. Por un lado, realizando un levantamiento de una zona determinada mediante GPS y por otro lado mediante fotogrametría aérea utilizando un vehículo aéreo no tripulado. Dicho estudio surge de la necesidad de obtener cartografía de los elementos que nos rodean y en nuestro caso, como primera toma de contacto, en el empleo de una nueva tecnología. Puesto que este requisito ha existido siempre, es evidente que en cada época el avance de las distintas metodologías y tecnologías para la producción cartográfica es mayor. En la actualidad, una de las técnicas más empleadas es la Fotogrametría. La idea de poder sobrevolar una extensión determinada para obtener imágenes y posteriormente poder recomponer el modelo para obtener la geometría y los elementos de dicha extensión hace que esta técnica sea muy eficaz. Hoy en día, las técnicas siguen avanzando y encontramos en el levantamiento mediante la utilización de vehículos aéreos no tripulados una de las mejores alternativas. Se trata de una herramienta muy potente en la realización de trabajos a grandes escalas (1:1000, 1:500). Comparado con métodos tradicionales, la fotogrametría aérea mediante vehículos aéreos no tripulados ofrece una gran cantidad de ventajas: · Mayor precisión: la alta resolución de las fotos tomadas permite obtener modelos digitales del terreno con mayor nivel de detalle. · Ahorro de tiempo: el levantamiento topográfico se puede completar en cuestión de horas, en comparación a métodos de topografía terrestre y fotogrametría tradicional. · Reducción de costos operativos: ya que levantamiento topográfico toma menos tiempo y menos recursos. · Mejoras en seguridad: el uso de una plataforma UAV puede eliminar o reducir la necesidad de acceder a zonas peligrosas. · Manejo de mayor información: fotografías verticales u oblicuas combinadas, levantamientos topográficos precisos, modelos digitales del terreno y análisis volumétrico. · Facilita la toma de decisiones: basada en información relevante y actualizada.

Structural basis of an embryonically lethal single Ala → Thr mutation in the vnd/NK-2 homeodomain

The structural and DNA binding behavior is described for an analog of the vnd/NK-2 homeodomain, which contains a single amino acid residue alanine to threonine replacement in position 35 of the homeodomain. Multidimensional nuclear magnetic resonance, circular dichroism, and electrophoretic gel retardation assays were carried out on recombinant 80-aa residue proteins that encompass the wild-type and mutant homeodomains. The mutant A35T vnd/NK-2 homeodomain is unable to adopt a folded conformation free in solution at temperatures down to −5°C in contrast to the behavior of the corresponding wild-type vnd/NK-2 homeodomain, which is folded into a functional three-dimensional structure below 25°C. The A35T vnd/NK-2 binds specifically to the vnd/NK-2 target DNA sequence, but with an affinity that is 50-fold lower than that of the wild-type homeodomain. Although the three-dimensional structure of the mutant A35T vnd/NK-2 in the DNA bound state shows characteristic helix–turn–helix behavior similar to that of the wild-type homeodomain, a notable structural deviation in the mutant A35T analog is observed for the amide proton of leucine-40. The wild-type homeodomain forms an unusual i,i-5 hydrogen bond with the backbone amide oxygen of residue 35. In the A35T mutant this amide proton resonance is shifted upfield by 1.27 ppm relative to the resonance frequency for the wild-type analog, thereby indicating a significant alteration of this i,i-5 hydrogen bond.

Bacterial resistance to vancomycin: Overproduction, purification, and characterization of VanC2 from Enterococcus casseliflavus as a d-Ala-d-Ser ligase

The VanC phenotype for clinical resistance of enterococci to vancomycin is exhibited by Enterococcus gallinarum and Enterococcus casseliflavus. Based on the detection of the cell precursor UDP-N-acetylmuramic acid pentapeptide intermediate terminating in d-Ala-d-Ser instead of d-Ala-d-Ala, it has been predicted that the VanC ligase would be a d-Ala-d-Ser rather than a d-Ala-d-Ala ligase. Overproduction of the E. casseliflavus ATCC 25788 vanC2 gene in Escherichia coli and its purification to homogeneity allowed demonstration of ATP-dependent d-Ala-d-Ser ligase activity. The kcat/Km2 (Km2 = Km for d-Ser or C-terminal d-Ala) ratio for d-Ala-d-Ser/d-Ala-d-Ala dipeptide formation is 270/0.69 for a 400-fold selection against d-Ala in the C-terminal position. VanC2 also has substantial d-Ala-d-Asn ligase activity (kcat/Km2 = 74 mM−1min−1).

Inhibition of ubiquitin/proteasome-dependent protein degradation by the Gly-Ala repeat domain of the Epstein–Barr virus nuclear antigen 1

The Epstein–Barr virus (EBV) encoded nuclear antigen (EBNA) 1 is expressed in latently infected B lymphocytes that persist for life in healthy virus carriers and is the only viral protein regularly detected in all EBV associated malignancies. The Gly-Ala repeat domain of EBNA1 was shown to inhibit in cis the presentation of major histocompatibility complex (MHC) class I restricted cytotoxic T cell epitopes from EBNA4. It appears that the majority of antigens presented via the MHC I pathway are subject to ATP-dependent ubiquitination and degradation by the proteasome. We have investigated the influence of the repeat on this process by comparing the degradation of EBNA1, EBNA4, and Gly-Ala containing EBNA4 chimeras in a cell-free system. EBNA4 was efficiently degraded in an ATP/ubiquitin/proteasome-dependent fashion whereas EBNA1 was resistant to degradation. Processing of EBNA1 was restored by deletion of the Gly-Ala domain whereas insertion of Gly-Ala repeats of various lengths and in different positions prevented the degradation of EBNA4 without appreciable effect on ubiquitination. Inhibition was also achieved by insertion of a Pro-Ala coding sequence. The results suggest that the repeat may affect MHC I restricted responses by inhibiting antigen processing via the ubiquitin/proteasome pathway. The presence of regularly interspersed Ala residues appears to be important for the effect.