Definición, diseño e implementación de métricas de usuario final sobre componentes web en el lado Front-end, mediante Google Analytics

La web ha evolucionado hacia la participación en la creación de contenido tanto por desarrolladores expertos como por usuarios finales sin un gran conocimiento en esta área. A pesar de que su uso es igual de válido y funcional, las diferencias entre la calidad de los productos desarrollados por ambos puede llegar a ser considerable. Esta característica se observa con mayor claridad cuando se analizan los web components. El trabajo consiste en el desarrollo de un entorno capaz de recoger las métricas de calidad de los componentes, basadas en la interacción con ellos por parte de los usuarios. A partir de las métricas obtenidas, se determinará su calidad para realizar una mejora de la misma, en función de las características valoradas. La selección de las métricas se realiza mediante un estudio de las características que definen a un componente, y permiten ser analizadas. Para poder llevar a cabo la construcción del portal, se ha descrito un prototipo capaz de proporcionar un sistema para permitir que los componentes intercambien información entre ellos. El modelo ha sido integrado en los componentes que se han de evaluar para obtener nuevas métricas sobre esta característica. Se ha desarrollado un dashboard que permite la interacción sin limitaciones de los usuarios con los componentes, facilitándoles un sistema para conectar componentes, utilizando para ello el sistema previamente descrito. Como conclusión del trabajo, se puede observar la necesidad de integrar los componentes web en un entorno real para poder determinar su calidad. Debido a que la calidad está determinada por los usuarios que consumen los componentes, se ha de contar con su opinión en la cuantificación de la misma.---ABSTRACT---Recently, the web has evolved to the collaboration between professional developers and end users with limited knowledge to create web content. Although both solutions are correct and functional, the differences in the quality between them can be appreciable. This feature is shown clearly when the web components are analyzed. The work is composed of the development of a virtual environment which is able to pick the quality measures of the components, based on the interaction between these components and the user. The measures are the starting point to decide the quality, and improve them with the rated measures. The measures selection is done through a study of the main features of a component. This selection can be analyzed. In order to create the website, a prototype has been specified to provide a system in which the components can be trade information between them. The interconnection model has been integrated in the components to evaluate. A dashboard has been developed to allow users interacting with the components without rules, making them possible connecting components through the model. The main conclusion of the work is the necessity of integrating web components in a real environment to decide their quality. Due to the fact that the quality is measured in terms of the rate of the users, it is a must to give them the main roles in the establishment of that quality.

Optimum sizing of bare-tape tethers for de-orbiting satellites at end of mission

De-orbiting satellites at end of mission would prevent generation of new space debris. A proposed de-orbit technology involves a bare conductive tape-tether, which uses neither propellant nor power supply while generating power for on-board use during de-orbiting. The present work shows how to select tape dimensions for a generic mission so as to satisfy requirements of very small tether-to-satellite mass ratio mt/MS and probability Nf of tether cut by small debris, while keeping de-orbit time tf short and product tf ×× tether length low to reduce maneuvers in avoiding collisions with large debris. Design is here discussed for particular missions (initial orbit of 720 km altitude and 63° and 92° inclinations, and 3 disparate MS values, 37.5, 375, and 3750 kg), proving it scalable. At mid-inclination and a mass-ratio of a few percent, de-orbit time takes about 2 weeks and Nf is a small fraction of 1%, with tape dimensions ranging from 1 to 6 cm, 10 to 54 μμm, and 2.8 to 8.6 km. Performance drop from middle to high inclination proved moderate: if allowing for twice as large mt/MS, increases are reduced to a factor of 4 in tf and a slight one in Nf, except for multi-ton satellites, somewhat more requiring because efficient orbital-motion-limited electron collection restricts tape-width values, resulting in tape length (slightly) increasing too.

Mejoras y extensiones en el Front-End de un modelo de predicción de acciones de un estudiante en un laboratorio virtual 3D

Este documento presenta las mejoras y las extensiones introducidas en la herramienta de visualización del modelo predictivo del comportamiento del estudiante o Student Behavior Predictor Viewer (SBPV), implementada en un trabajo anterior. El modelo predictivo del comportamiento del estudiante es parte de un sistema inteligente de tutoría, y se construye a partir de los registros de actividad de los estudiantes en un laboratorio virtual 3D, como el Laboratorio Virtual de Biotecnología Agroforestal, implementado en un trabajo anterior, y cuyos registros de actividad de los estudiantes se han utilizado para validar este trabajo fin de grado. El SBPV es una herramienta para visualizar una representación gráfica 2D del grafo extendido asociado con cualquiera de los clusters del modelo predictivo del estudiante. Además de la visualización del grafo extendido, el SBPV controla la navegación a través del grafo por medio del navegador web. Más concretamente, el SBPV permite al usuario moverse a través del grafo, ampliar o reducir el zoom del gráfico o buscar un determinado estado. Además, el SBPV también permite al usuario modificar el diseño predeterminado del grafo en la pantalla al cambiar la posición de los estados con el ratón. Como parte de este trabajo fin de grado, se han corregido errores existentes en la versión anterior y se han introducido una serie de mejoras en el rendimiento y la usabilidad. En este sentido, se han implementado nuevas funcionalidades, tales como la visualización del modelo de comportamiento de cada estudiante individualmente o la posibilidad de elegir el método de clustering para crear el modelo predictivo del estudiante; así como ha sido necesario rediseñar la interfaz de usuario cambiando el tipo de estructuras gráficas con que se muestran los elementos del modelo y mejorando la visualización del grafo al interaccionar el usuario con él. Todas estas mejoras se explican detenidamente en el presente documento.---ABSTRACT---This document presents the improvements and extensions made to the visualization tool Student Behavior Predictor Viewer (SBPV), implemented in a previous job. The student behavior predictive model is part of an intelligent tutoring system, and is built from the records of students activity in a 3D virtual laboratory, like the “Virtual Laboratory of Agroforestry Biotechnology” implemented in a previous work, and whose records of students activity have been used to validate this final degree work. The SBPV is a tool for visualizing a 2D graphical representation of the extended graph associated with any of the clusters of the student predictive model. Apart from visualizing the extended graph, the SBPV supports the navigation across the graph by means of desktop devices. More precisely, the SBPV allows user to move through the graph, to zoom in/out the graphic or to locate a given state. In addition, the SBPV also allows user to modify the default layout of the graph on the screen by changing the position of the states by means of the mouse. As part of this work, some bugs of the previous version have been fixed and some enhancements have been implemented to improve the performance and the usability. In this sense, we have implemented new features, such as the display of the model behavior of only one student or the possibility of selecting the clustering method to create the student predictive model; as well as it was necessary to redesign the user interface changing the type of graphic structures that show model elements and improving the rendering of the graph when the user interacts with it. All these improvements are explained in detail in the next sections.

Functional copies of a human gene can be directly isolated by transformation-associated recombination cloning with a small 3′ end target sequence

Unique, small sequences (sequence tag sites) have been identified at the 3′ ends of most human genes that serve as landmarks in genome mapping. We investigated whether a single copy gene could be isolated directly from total human DNA by transformation-associated recombination (TAR) cloning in yeast using a short, 3′ unique target. A TAR cloning vector was constructed that, when linearized, contained a small amount (381 bp) of 3′ hypoxanthine phosphoribosyltransferase (HPRT) sequence at one end and an 189-bp Alu repeat at the other end. Transformation with this vector along with human DNA led to selective isolations of the entire HPRT gene as yeast artificial chromosomes (YACs) that extended from the 3′ end sequence to various Alu positions as much as 600 kb upstream. These YACs were retrofitted with a NeoR and a bacterial artificial chromosome (BAC) sequence to transfer the YACs to bacteria and subsequently the BACs to mouse cells by using a Neo selection. Most of the HPRT isolates were functional, demonstrating that TAR cloning retains the functional integrity of the isolated material. Thus, this modified version of TAR cloning, which we refer to as radial TAR cloning, can be used to isolate large segments of the human genome accurately and directly with only a small amount of sequence information.

Breakers of advanced glycation end products restore large artery properties in experimental diabetes

Glucose and other reducing sugars react with proteins by a nonenzymatic, posttranslational modification process called nonenzymatic glycation. The formation of advanced glycation end products (AGEs) on connective tissue and matrix components accounts largely for the increase in collagen crosslinking that accompanies normal aging and which occurs at an accelerated rate in diabetes, leading to an increase in arterial stiffness. A new class of AGE crosslink “breakers” reacts with and cleaves these covalent, AGE-derived protein crosslinks. Treatment of rats with streptozotocin-induced diabetes with the AGE-breaker ALT-711 for 1–3 weeks reversed the diabetes-induced increase of large artery stiffness as measured by systemic arterial compliance, aortic impedance, and carotid artery compliance and distensibility. These findings will have considerable implications for the treatment of patients with diabetes-related complications and aging.

Epitopes close to the apolipoprotein B low density lipoprotein receptor-binding site are modified by advanced glycation end products

Advanced glycation end products (AGEs) are thought to contribute to the abnormal lipoprotein profiles and increased risk of cardiovascular disease of patients with diabetes and renal failure, in part by preventing apolipoprotein B (apoB)-mediated cellular uptake of low density lipoproteins (LDL) by LDL receptors (LDLr). It has been proposed that AGE modification at one site in apoB, almost 1,800 residues from the putative apoB LDLr-binding domain, may be sufficient to induce an apoB conformational change that prevents binding to the LDLr. To further explore this hypothesis, we used 29 anti-human apoB mAbs to identify other potential sites on apoB that may be modified by in vitro advanced glycation of LDL. Glycation of LDL caused a time-dependent decrease in its ability to bind to the LDLr and in the immunoreactivity of six distinct apoB epitopes, including two that flank the apoB LDLr-binding domain. ApoB appears to be modified at multiple sites by these criteria, as the loss of glycation-sensitive epitopes was detected on both native glycated LDL and denatured, delipidated glycated apoB. Moreover, residues directly within the putative apoB LDLr-binding site are not apparently modified in glycated LDL. We propose that the inability of LDL modified by AGEs to bind to the LDLr is caused by modification of residues adjacent to the putative LDLr-binding site that were undetected by previous immunochemical studies. AGE modification either eliminates the direct participation of the residues in LDLr binding or indirectly alters the conformation of the apoB LDLr-binding site.

The “Spot 14” gene resides on the telomeric end of the 11q13 amplicon and is expressed in lipogenic breast cancers: Implications for control of tumor metabolism

Enhanced long chain fatty acid synthesis may occur in breast cancer, where it is necessary for tumor growth and predicts a poor prognosis. “Spot 14” (S14) is a carbohydrate- and thyroid hormone-inducible nuclear protein specific to liver, adipose, and lactating mammary tissues that functions to activate genes encoding the enzymes of fatty acid synthesis. Amplification of chromosome region 11q13, where the S14 gene (THRSP) resides, also predicts a poor prognosis in breast tumors. We localized the S14 gene between markers D11S906 and D11S937, at the telomeric end of the amplified region at 11q13, and found that it was amplified and expressed in breast cancer-derived cell lines. Moreover, concordant expression of S14 and a key lipogenic enzyme (acetyl-CoA carboxylase) in a panel of primary breast cancer specimens strongly supported a role for S14 as a determinant of tumor lipid metabolism. S14 expression provides a pathophysiological link between two prognostic indicators in breast cancer: enhanced lipogenesis and 11q13 amplification.

Mammalian capping enzyme binds RNA and uses protein tyrosine phosphatase mechanism

Mammalian capping enzymes are bifunctional proteins with both RNA 5′-triphosphatase and guanylyltransferase activities. The N-terminal 237-aa triphosphatase domain contains (I/V)HCXXGXXR(S/T)G, a sequence corresponding to the conserved active-site motif in protein tyrosine phosphatases (PTPs). Analysis of point mutants of mouse RNA 5′-triphosphatase identified the motif Cys and Arg residues and an upstream Asp as required for activity. Like PTPs, this enzyme was inhibited by iodoacetate and VO43− and independent of Mg2+, providing additional evidence for phosphate removal from RNA 5′ ends by a PTP-like mechanism. The full-length, 597-aa mouse capping enzyme and the C-terminal guanylyltransferase fragment (residues 211–597), unlike the triphosphatase domain, bound poly (U) and were nuclear in transfected cells. RNA binding was increased by GTP, and a guanylylation-defective, active-site mutant was not affected. Ala substitution at positions required for the formation of the enzyme-GMP capping intermediate (R315, R530, K533, or N537) also eliminated poly (U) binding, while proteins with conservative substitutions at these sites retained binding but not guanylyltransferase activity. These results demonstrate that the guanylyltransferase domain of mammalian capping enzyme specifies nuclear localization and RNA binding. Association of capping enzyme with nascent transcripts may act in synergy with RNA polymerase II binding to ensure 5′ cap formation.

Rap1 protein regulates telomere turnover in yeast

Telomere length is maintained through a dynamic balance between addition and loss of the terminal telomeric DNA. Normal telomere length regulation requires telomerase as well as a telomeric protein–DNA complex. Previous work has provided evidence that in the budding yeasts Kluyveromyces lactis and Saccharomyces cerevisiae, the telomeric double-stranded DNA binding protein Rap1p negatively regulates telomere length, in part by nucleating, by its C-terminal tail, a higher-order DNA binding protein complex that presumably limits access of telomerase to the chromosome end. Here we show that in K. lactis, truncating the Rap1p C-terminal tail (Rap1p-ΔC mutant) accelerates telomeric repeat turnover in the distal region of the telomere. In addition, combining the rap1-ΔC mutation with a telomerase template mutation (ter1-kpn), which directs the addition of mutated telomeric DNA repeats to telomeres, synergistically caused an immediate loss of telomere length regulation. Capping of the unregulated telomeres of these double mutants with functionally wild-type repeats restored telomere length control. We propose that the rate of terminal telomere turnover is controlled by Rap1p specifically through its interactions with the most distal telomeric repeats.

Major histocompatibility complex class I-restricted T cells are required for all but the end stages of diabetes development in nonobese diabetic mice and use a prevalent T cell receptor α chain gene rearrangement

Nonobese diabetic (NOD) mice develop insulin-dependent diabetes mellitus due to autoimmune T lymphocyte-mediated destruction of pancreatic β cells. Although both major histocompatibility complex class I-restricted CD8+ and class II-restricted CD4+ T cell subsets are required, the specific role each subset plays in the pathogenic process is still unclear. Here we show that class I-dependent T cells are required for all but the terminal stages of autoimmune diabetes development. To characterize the diabetogenic CD8+ T cells responsible, we isolated and propagated in vitro CD8+ T cells from the earliest insulitic lesions of NOD mice. They were cytotoxic to NOD islet cells, restricted to H-2Kd, and showed a diverse T cell receptor β chain repertoire. In contrast, their α chain repertoire was more restricted, with a recurrent amino acid sequence motif in the complementarity-determining region 3 loop and a prevalence of Vα17 family members frequently joined to the Jα42 gene segment. These results suggest that a number of the CD8+ T cells participating in the initial phase of autoimmune β cell destruction recognize a common structural component of Kd/peptide complexes on pancreatic β cells, possibly a single peptide.

Rates of ubiquitin conjugation increase when muscles atrophy, largely through activation of the N-end rule pathway

The rapid loss of muscle mass that accompanies many disease states, such as cancer or sepsis, is primarily a result of increased protein breakdown in muscle, and several observations have suggested an activation of the ubiquitin–proteasome system. Accordingly, in extracts of atrophying muscles from tumor-bearing or septic rats, rates of 125I-ubiquitin conjugation to endogenous proteins were found to be higher than in control extracts. On the other hand, in extracts of muscles from hypothyroid rats, where overall proteolysis is reduced below normal, the conjugation of 125I-ubiquitin to soluble proteins decreased by 50%, and treatment with triiodothyronine (T3) restored ubiquitination to control levels. Surprisingly, the N-end rule pathway, which selectively degrades proteins with basic or large hydrophobic N-terminal residues, was found to be responsible for most of these changes in ubiquitin conjugation. Competitive inhibitors of this pathway that specifically block the ubiquitin ligase, E3α, suppressed most of the increased ubiquitin conjugation in the muscle extracts from tumor-bearing and septic rats. These inhibitors also suppressed ubiquitination in normal extracts toward levels in hypothyroid extracts, which showed little E3α-dependent ubiquitination. Thus, the inhibitors eliminated most of the differences in ubiquitination under these different pathological conditions. Moreover, 125I-lysozyme, a model N-end rule substrate, was ubiquitinated more rapidly in extracts from tumor-bearing and septic rats, and more slowly in those from hypothyroid rats, than in controls. Thus, the rate of ubiquitin conjugation increases in atrophying muscles, and these hormone- and cytokine-dependent responses are in large part due to activation of the N-end rule pathway.

Measurements of attractive forces between proteins and end-grafted poly(ethylene glycol) chains

The surface force apparatus was used to measure directly the molecular forces between streptavidin and lipid bilayers displaying grafted Mr 2,000 poly(ethylene glycol) (PEG). These measurements provide direct evidence for the formation of relatively strong attractive forces between PEG and protein. At low compressive loads, the forces were repulsive, but they became attractive when the proteins were pressed into the polymer layer at higher loads. The adhesion was sufficiently robust that separation of the streptavidin and PEG uprooted anchored polymer from the supporting membrane. These interactions altered the properties of the grafted chains. After the onset of the attraction, the polymer continued to bind protein for several hours. The changes were not due to protein denaturation. These data demonstrate directly that the biological activity of PEG is not due solely to properties of simple polymers such as the excluded volume. It is also coupled to the competitive interactions between solvent and other materials such as proteins for the chain segments and to the ability of this material to adopt higher order intrachain structures.

Phylogeny of mRNA capping enzymes

The m7GpppN cap structure of eukaryotic mRNA is formed cotranscriptionally by the sequential action of three enzymes: RNA triphosphatase, RNA guanylyltransferase, and RNA (guanine-7)-methyltransferase. A multifunctional polypeptide containing all three active sites is encoded by vaccinia virus. In contrast, fungi and Chlorella virus encode monofunctional guanylyltransferase polypeptides that lack triphosphatase and methyltransferase activities. Transguanylylation is a two-stage reaction involving a covalent enzyme-GMP intermediate. The active site is composed of six protein motifs that are conserved in order and spacing among yeast and DNA virus capping enzymes. We performed a structure–function analysis of the six motifs by targeted mutagenesis of Ceg1, the Saccharomyces cerevisiae guanylyltransferase. Essential acidic, basic, and aromatic functional groups were identified. The structural basis for covalent catalysis was illuminated by comparing the mutational results with the crystal structure of the Chlorella virus capping enzyme. The results also allowed us to identify the capping enzyme of Caenorhabditis elegans. The 573-amino acid nematode protein consists of a C-terminal guanylyltransferase domain, which is homologous to Ceg1 and is strictly conserved with respect to all 16 amino acids that are essential for Ceg1 function, and an N-terminal phosphatase domain that bears no resemblance to the vaccinia triphosphatase domain but, instead, has strong similarity to the superfamily of protein phosphatases that act via a covalent phosphocysteine intermediate.

Dictyostelium myosin II null mutant can still cap Con A receptors

Cross-linked antigens on the surface of a motile cell cap at the trailing end of the cell. In Dictyostelium discoideum, myosin II null mutants have previously been reported to be unable to cap Con A receptors, although they are able to locomote. This finding implicated myosin II as an essential component of the capping mechanism, although not of the machinery for locomotion. Here we show that myosin II null mutants do cap Con A receptors, albeit less efficiently than does wild type. This shows that cap formation is not absolutely dependent on myosin II and that a close mechanistic relationship between capping, particle movement, and cell migration may still exist.

Light-induced exposure of the cytoplasmic end of transmembrane helix seven in rhodopsin

A key step in signal transduction in the visual cell is the light-induced conformational change of rhodopsin that triggers the binding and activation of the guanine nucleotide-binding protein. Site-directed mAbs against bovine rhodopsin were produced and used to detect and characterize these conformational changes upon light activation. Among several antibodies that bound exclusively to the light-activated state, an antibody (IgG subclass) with the highest affinity (Ka ≈ 6 × 10−9 M) was further purified and characterized. The epitope of this antibody was mapped to the amino acid sequence 304–311. This epitope extends from the central region to the cytoplasmic end of the seventh transmembrane helix and incorporates a part of a highly conserved NPXXY motif, a critical region for signaling and agonist-induced internalization of several biogenic amine and peptide receptors. In the dark state, no binding of the antibody to rhodopsin was detected. Accessibility of the epitope to the antibody correlated with formation of the metarhodopsin II photointermediate and was reduced significantly at the metarhodopsin III intermediate. Further, incubation of the antigen–antibody complex with 11-cis-retinal failed to regenerate the native rhodopsin chromophore. These results suggest significant and reversible conformational changes in close proximity to the cytoplasmic end of the seventh transmembrane helix of rhodopsin that might be important for folding and signaling.