Overall vs. local mechanical behaviour of ferritic austenitic steel joints made by laser welding

The present investigation addresses the overall and local mechanical performance of dissimilar joints of low carbon steel (CS) and stainless Steel (SS) thin sheets achieved by laser welding in case of heat source displacement from the weld gap centreline towards CS. Welding was performed on a Nd:YAG laser DY033 (3300 W) in a continuos wave (CW), keyhole mode. The tensile behavior of the joint different zones assessed by using a video-image based system (VIC-2D) reveals that the residual stress field, together with the positive difference in yield between the weld metal and the base materials protects the joint from being plastically deformed. The tensile loadings of flat transverse specimens generate the strain localization and failure in CS, far away from the weld.

Detección de marcas para realidad aumentada sobre dispositivos móviles

Este documento es una guía para el desarrollo de una aplicación para dispositivos móviles en Android. Dicha aplicación combina las técnicas de visión por computador para calibrar la cámara del dispositivo y localizar un elemento en el espacio en base a esos los parámetros calculados en la calibración. El diseño de la aplicación incluye las decisiones sobre la forma en que se reciben los inputs de la aplicación, que patrones se utilizan en la calibración y en la localización y como se muestran los resultados finales al usuario. También incluye un diagrama de flujo de información que representa el tránsito de esta entre los diferentes módulos. La implementación comienza con la configuración de un entorno para desarrollar aplicaciones con parte nativa en Android, después comenta el código de la aplicación paso por paso incluyendo comentarios sobre los archivos adicionales necesarios para la compilación y finalmente detalla los archivos dedicados a la interfaz. Los experimentos incluyen una breve descripción sobre cómo interpretar los resultados seguidos de una serie de imágenes tomadas de la aplicación con diferentes localizaciones del patrón. En la entrega se incluye también un video. En el capítulo de resultados y conclusiones podemos encontrar observaciones sobre el desarrollo de la práctica, opiniones sobre su utilidad, y posibles mejoras.---ABSTRACT---This document is a guide that describes the development of and application for mobile devices in Android OS. The application combines computer vision techniques to calibrate the device camera and locate an element in the real world based on the parameters of the calibration The design of the application includes the decisions over the way that the application receives its input data, the patterns used in the calibration and localization and how the results are shown to the user. It also includes a flow chart that describes how the information travels along the application modules. The development begins with the steps necessary to configure the environment to develop native Android applications, then it explains the code step by step, including commentaries on the additional files necessary to build the application and details the files of the user interface. The experiments chapter explains the way the results are shown in the experiments before showing samples of different pattern localizations. There is also a video attached. In the conclusions chapter we can find observations on the development of the TFG, opinions about its usefulness, and possibilities of improvement in the future.

Some advances in extensive bridge monitoring using low cost dynamic characterization

Dynamic measurements will become a standard for bridge monitoring in the near future. This fact will produce an important cost reduction for maintenance. US Administration has a long term intensive research program in order to diminish the estimated current maintenance cost of US$7 billion per year over 20 years. An optimal intervention maintenance program demands a historical dynamical record, as well as an updated mathematical model of the structure to be monitored. In case that a model of the structure is not actually available it is possible to produce it, however this possibility does not exist for missing measurement records from the past. Current acquisition systems to monitor structures can be made more efficient by introducing the following improvements, under development in the Spanish research Project “Low cost bridge health monitoring by ambient vibration tests using wireless sensors”: (a) a complete wireless system to acquire sensor data, (b) a wireless system that permits the localization and the hardware identification of the whole sensor system. The applied localization system has been object of a recent patent, and (c) automatization of the modal identification process, aimed to diminish human intervention. This system is assembled with cheap components and allows the simultaneous use of a large number of sensors at a low placement cost. The engineer’s intervention is limited to the selection of sensor positions, probably based on a preliminary FE analysis. In case of multiple setups, also the position of a number of fixed reference sensors has to be decided. The wireless localization system will obtain the exact coordinates of all these sensors positions. When the selection of optimal positions is difficult, for example because of the lack of a proper FE model, this can be compensated by using a higher number of measuring (also reference) points. The described low cost acquisition system allows the responsible bridge administration to obtain historical dynamic identification records at reasonable costs that will be used in future maintenance programs. Therefore, due to the importance of the baseline monitoring record of a new bridge, a monitoring test just after its construction should be highly recommended, if not compulsory.

El sistema de escuelas primarias de Juan O'Gorman, arquitecto : modernidad y eficiencia

El proceso de proyecto, llevado a cabo por Juan O’Gorman, para proyectar y construir más de veinticinco Escuelas Primarias en el Distrito Federal en menos de seis meses en 1932, necesariamente debió regirse por una metodología que permitiera conseguirlo. Esta investigación plantea la existencia de un método de proyecto que engloba toda la arquitectura de la etapa funcionalista de O’Gorman. Se considera la metodología, denominada en esta tesis Sistema Modular de las Escuelas Primarias, como un procedimiento proyectual evolutivo, conseguido a través de una investigación y experimentación continua. Inicia en 1929 con la realización de dos proyectos de vivienda particular, continúa con la génesis de un esquema de modulación que surge en unos proyectos de vivienda colectiva, se perfecciona en los proyectos de Escuelas Primarias del D.F. introduciendo criterios de estandarización y culmina con la adaptación a diversas circunstancias geométricas y climáticas que define la ubicación, en los casos concretos de la Escuela Vocacional Industrial de Tolsá y Tresguerras y la Escuela Primaria de Tampico en 1934. Este planteamiento destaca el valor de la obra funcionalista de aquel joven arquitecto, no sólo por lo novedoso que este nuevo lenguaje supuso en el México neo-colonial de los años 30, sino especialmente por la importancia que significó para la arquitectura mexicana contemporánea, la sistematización de procesos de proyecto -modulación y estandarización- para abordar una arquitectura inexistente en la época -la arquitectura social- de manera moderna y eficiente. La metodología empleada en la tesis implica la catalogación y mapeado de las Escuelas Primarias del D.F., lo cual aporta documentación original inédita. ABSTRACT The project process, carried out by Juan O’Gorman, to design and build more than twenty five elementary schools in less than six months in Mexico City in 1932, must have necessarily been governed by a thorough methodology in order to achieve this. This research posits the existence of a project method that encompasses all the architecture of O’Gorman’s functionalist stage. The methodology, known as “Modular System of Elementary Schools” in this thesis, is regarded as an evolutionary proyectual process, achieved through continuous research and experimentation. This methodology, which started in 1929 with the completion of two projects of private dwellings, continued with the creation of a modulation scheme arising from collective housing projects, later perfected in the Elementary School projects of Mexico City by introducing standardization criteria. Finally, it culminated in the adaptation to various geometric and climatic circumstances defined by the location, in the specific cases of the Industrial Vocational School of Tolsá and Tresguerras and Tampico’s Elementary School in 1934. This approach highlights the value of the young architect’s functionalist work, not only for the novelty of this new language in the neo-colonial Mexico of the 1930s, but especially because of the importance it meant to contemporary Mexican architecture, the systematization of project processes, -modulation and standardization- and addressing a nonexistent ‘social architecture’ at the time in a modern and efficient way. The methodology used in this thesis involves cataloging and mapping O’Gorman’s Elementary Schools in Mexico City, which provides unprecedented original documentation.

A third member of the synapsin gene family

Synapsins are a family of neuron-specific synaptic vesicle-associated phosphoproteins that have been implicated in synaptogenesis and in the modulation of neurotransmitter release. In mammals, distinct genes for synapsins I and II have been identified, each of which gives rise to two alternatively spliced isoforms. We have now cloned and characterized a third member of the synapsin gene family, synapsin III, from human DNA. Synapsin III gives rise to at least one protein isoform, designated synapsin IIIa, in several mammalian species. Synapsin IIIa is associated with synaptic vesicles, and its expression appears to be neuron-specific. The primary structure of synapsin IIIa conforms to the domain model previously described for the synapsin family, with domains A, C, and E exhibiting the highest degree of conservation. Synapsin IIIa contains a novel domain, termed domain J, located between domains C and E. The similarities among synapsins I, II, and III in domain organization, neuron-specific expression, and subcellular localization suggest a possible role for synapsin III in the regulation of neurotransmitter release and synaptogenesis. The human synapsin III gene is located on chromosome 22q12–13, which has been identified as a possible schizophrenia susceptibility locus. On the basis of this localization and the well established neurobiological roles of the synapsins, synapsin III represents a candidate gene for schizophrenia.

Neuronal coincidence detection by voltage-sensitive electrical synapses

Coincidence detection is important for functions as diverse as Hebbian learning, binaural localization, and visual attention. We show here that extremely precise coincidence detection is a natural consequence of the normal function of rectifying electrical synapses. Such synapses open to bidirectional current flow when presynaptic cells depolarize relative to their postsynaptic targets and remain open until well after completion of presynaptic spikes. When multiple input neurons fire simultaneously, the synaptic currents sum effectively and produce a large excitatory postsynaptic potential. However, when some inputs are delayed relative to the rest, their contributions are reduced because the early excitatory postsynaptic potential retards the opening of additional voltage-sensitive synapses, and the late synaptic currents are shunted by already opened junctions. These mechanisms account for the ability of the lateral giant neurons of crayfish to sum synchronous inputs, but not inputs separated by only 100 μsec. This coincidence detection enables crayfish to produce reflex escape responses only to very abrupt mechanical stimuli. In light of recent evidence that electrical synapses are common in the mammalian central nervous system, the mechanisms of coincidence detection described here may be widely used in many systems.

Postsynaptic clustering of γ-aminobutyric acid type A receptors by the γ3 subunit in vivo

Synaptic localization of γ-aminobutyric acid type A (GABAA) receptors is a prerequisite for synaptic inhibitory function, but the mechanism by which different receptor subtypes are localized to postsynaptic sites is poorly understood. The γ2 subunit and the postsynaptic clustering protein gephyrin are required for synaptic localization and function of major GABAA receptor subtypes. We now show that transgenic overexpression of the γ3 subunit in γ2 subunit-deficient mice restores benzodiazepine binding sites, benzodiazepine-modulated whole cell currents, and postsynaptic miniature currents, suggesting the formation of functional, postsynaptic receptors. Moreover, the γ3 subunit can substitute for γ2 in the formation of GABAA receptors that are synaptically clustered and colocalized with gephyrin in vivo. These clusters were formed even in brain regions devoid of endogenous γ3 subunit, indicating that the factors present for clustering of γ2 subunit-containing receptors are sufficient to cluster γ3 subunit-containing receptors. The GABAA receptor and gephyrin-clustering properties of the ectopic γ3 subunit were also observed for the endogenous γ3 subunit, but only in the absence of the γ2 subunit, suggesting that the γ3 subunit is at a competitive disadvantage with the γ2 subunit for clustering of postsynaptic GABAA receptors in wild-type mice.

Activity-dependent regulation of synaptic clustering in a hippocampal culture system

Currently, there is a limited understanding of the factors that influence the localization and density of individual synapses in the central nervous system. Here we have studied the effects of activity on synapse formation between hippocampal dentate granule cells and CA3 pyramidal neurons in culture, taking advantage of FM1–43 as a fluorescent marker of synaptic boutons. We observed an early tendency for synapses to group together, quickly followed by the appearance of synaptic clusters on dendritic processes. These events were strongly influenced by N-methyl-d-aspartic acid receptor- and cyclic AMP-dependent signaling. The microstructure and localization of the synaptic clusters resembled that found in hippocampus, at mossy fiber synapses of stratum lucidum. Activity-dependent clustering of synapses represents a means for synaptic targeting that might contribute to synaptic organization in the brain.

Direct isolation of human transcribed sequences from yeast artificial chromosomes through the application of RNA fingerprinting

The identification of cDNA clones from genomic regions known to contain human genes is usually the rate-limiting factor in positional cloning strategies. We demonstrate here that human genes present on yeast artificial chromosomes (YACs) are transcribed in yeast host cells. We have used the arbitrarily primed RNA (RAP) fingerprinting method to identify human-specific, transcribed sequences from YACs located in the 13q12 chromosome region. By comparing the RAP fingerprints generated using defined, arbitrary primers from various fragmented YACs, megaYACs, and host yeast, we were able to identify and map 20 products transcribed from the human YAC inserts. This method, therefore, permits the simultaneous isolation and mapping of novel expressed sequences directly from whole YACs.

Cell death induction by the acute promyelocytic leukemia-specific PML/RARα fusion protein

PML/RARα is the abnormal protein product generated by the acute promyelocytic leukemia-specific t(15;17). Expression of PML/RARα in hematopoietic precursor cell lines induces block of differentiation and promotes survival. We report here that PML/RARα has a potent growth inhibitory effect on all nonhematopoietic cell lines and on the majority of the hematopoietic cell lines tested. Inducible expression of PML/RARα in fibroblasts demonstrated that the basis for the growth suppression is induction of cell death. Deletion of relevant promyelocytic leukemia (PML) and retinoic acid receptor (RARα) domains within the fusion protein revealed that its growth inhibitory effect depends on the integrity of the PML aminoterminal region (RING, B1, B2, and coiled coil regions) and the RARα DNA binding region. Analysis of the nuclear localization of the same PML/RARα deletion mutants by immunofluorescence and cell fractionation revealed that the biological activity of the fusion protein correlates with its microspeckled localization and its association to the nuclear matrix. The PML aminoterminal region, but not the RARα zinc fingers, is required for the proper nuclear localization of PML/RARα. We propose that the matrix-associated microspeckles are the active sites of PML/RARα and that targeting of RARα sequences to this specific nuclear subdomain through PML sequences is crucial to the activity of the fusion protein on survival regulation.

Nuclear punctate distribution of ALL-1 is conferred by distinct elements at the N terminus of the protein

The ALL-1 gene positioned at 11q23 is directly involved in human acute leukemia either through a variety of chromosome translocations or by partial tandem duplications. ALL-1 is the human homologue of Drosophila trithorax which plays a critical role in maintaining proper spatial and temporal expression of the Antennapedia-bithorax homeotic genes determining the fruit fly’s body pattern. Utilizing specific antibodies, we found that the ALL-1 protein distributes in cultured cells in a nuclear punctate pattern. Several chimeric ALL-1 proteins encoded by products of the chromosome translocations and expressed in transfected cells showed similar speckles. Dissection of the ALL-1 protein identified within its ≈1,100 N-terminal residues three polypeptides directing nuclear localization and at least two main domains conferring distribution in dots. The latter spanned two short sequences conserved with TRITHORAX. Enforced nuclear expression of other domains of ALL-1, such as the PHD (zinc) fingers and the SET motif, resulted in uniform nonpunctate patterns. This indicates that positioning of the ALL-1 protein in subnuclear structures is mediated via interactions of ALL-1 N-terminal elements. We suggest that the speckles represent protein complexes which contain multiple copies of the ALL-1 protein and are positioned at ALL-1 target sites on the chromatin. Therefore, the role of the N-terminal portion of ALL-1 is to direct the protein to its target genes.

The Stem-Loop Binding Protein (SLBP1) Is Present in Coiled Bodies of the Xenopus Germinal Vesicle

The stem-loop binding protein (SLBP1) binds the 3′ stem-loop of histone pre-mRNA and is required for efficient processing of histone transcripts in the nucleus. We examined the localization of SLBP1 in the germinal vesicle of Xenopus laevis oocytes. In spread preparations of germinal vesicle contents, an anti-SLBP1 antibody stained coiled bodies and specific chromosomal loci, including terminal granules, axial granules, and some loops. After injection of myc-tagged SLBP1 transcripts into the oocyte cytoplasm, newly translated myc-SLBP1 protein was detectable in coiled bodies within 4 h and in terminal and axial granules by 8 h. To identify the region(s) of SLBP1 necessary for subnuclear localization, we subcloned various parts of the SLBP1 cDNA and injected transcripts of these into the cytoplasm of oocytes. We determined that 113 amino acids at the carboxy terminus of SLBP1 are sufficient for coiled body localization and that disruption of a previously defined RNA-binding domain did not alter this localization. Coiled bodies also contain the U7 small nuclear ribonucleoprotein particle (snRNP), which participates in cleavage of the 3′ end of histone pre-mRNA. The colocalization of SLBP1 and the U7 snRNP in the coiled body suggests coordinated control of their functions, perhaps through a larger histone-processing particle. Some coiled bodies are attached to the lampbrush chromosomes at the histone gene loci, consistent with the view that coiled bodies in the oocyte recruit histone-processing factors to the sites of histone pre-mRNA transcription. The non-histone chromosomal sites at which SLBP1 is found include the genes coding for 5 S rRNA, U1 snRNA, and U2 snRNA, suggesting a wider role for SLBP1 in the biosynthesis of small non-spliced RNAs.

Effect of Carbohydrate Position on Lysosomal Transport of Procathepsin L

To study the role of carbohydrate in lysosomal protein transport, we engineered two novel glycosylation signals (Asn-X-Ser/Thr) into the cDNA of human procathepsin L, a lysosomal acid protease. We constructed six mutant cDNAs encoding glycosylation signals at mutant sites Asn-138, Asn-175, or both sites together, in the presence or absence of the wild-type Asn-204 site. We stably transfected wild-type and mutant cDNAs into NIH3T3 mouse fibroblasts and then used species-specific antibodies to determine the glycosylation status, phosphorylation, localization, and transport kinetics of recombinant human procathepsin L containing one, two, or three glycosylation sites. Both novel glycosylation sites were capable of being glycosylated, although Asn-175 was utilized only 30–50% of the time. Like the wild-type glycosylation at Asn-204, carbohydrates at Asn-138 and Asn-175 were completely sensitive to endoglycosidase H, and they were phosphorylated. Mutant proteins containing two carbohydrates were capable of being delivered to lysosomes, but there was not a consistent relationship between the efficiency of lysosomal delivery and carbohydrate content of the protein. Pulse-chase labeling revealed a unique biosynthetic pattern for proteins carrying the Asn-175 glycosylation sequence. Whereas wild-type procathepsin L and mutants bearing carbohydrate at Asn-138 appeared in lysosomes by about 60 min, proteins with carbohydrate at Asn-175 were processed to a lysosome-like polypeptide within 15 min. Temperature shift, brefeldin A, and NH4Cl experiments suggested that the rapid processing did not occur in the endoplasmic reticulum and that Asn-175 mutants could interact with the mannose 6-phosphate receptor. Taken together, our results are consistent with the interpretation that Asn-175 carbohydrate confers rapid transport to lysosomes. We may have identified a recognition domain in procathepsin L that is important for its interactions with the cellular transport machinery.

A Role for the GSG Domain in Localizing Sam68 to Novel Nuclear Structures in Cancer Cell Lines

The GSG (GRP33, Sam68, GLD-1) domain is a protein module found in an expanding family of RNA-binding proteins. The numerous missense mutations identified genetically in the GSG domain support its physiological role. Although the exact function of the GSG domain is not known, it has been shown to be required for RNA binding and oligomerization. Here it is shown that the Sam68 GSG domain plays a role in protein localization. We show that Sam68 concentrates into novel nuclear structures that are predominantly found in transformed cells. These Sam68 nuclear bodies (SNBs) are distinct from coiled bodies, gems, and promyelocytic nuclear bodies. Electron microscopic studies show that SNBs are distinct structures that are enriched in phosphorus and nitrogen, indicating the presence of nucleic acids. A GFP-Sam68 fusion protein had a similar localization as endogenous Sam68 in HeLa cells, diffusely nuclear with two to five SNBs. Two other GSG proteins, the Sam68-like mammalian proteins SLM-1 and SLM-2, colocalized with endogenous Sam68 in SNBs. Different GSG domain missense mutations were investigated for Sam68 protein localization. Six separate classes of cellular patterns were obtained, including exclusive SNB localization and association with microtubules. These findings demonstrate that the GSG domain is involved in protein localization and define a new compartment for Sam68, SLM-1, and SLM-2 in cancer cell lines.

A potential role for the nuclear factor of activated T cells family of transcriptional regulatory proteins in adipogenesis

NFAT (nuclear factor of activated T cells) is a family of transcription factors implicated in the control of cytokine and early immune response gene expression. Recent studies have pointed to a role for NFAT proteins in gene regulation outside of the immune system. Herein we demonstrate that NFAT proteins are present in 3T3-L1 adipocytes and, upon fat cell differentiation, bind to and transactivate the promoter of the adipocyte-specific gene aP2. Further, fat cell differentiation is inhibited by cyclosporin A, a drug shown to prevent NFAT nuclear localization and hence function. Thus, these data suggest a role for NFAT transcription factors in the regulation of the aP2 gene and in the process of adipocyte differentiation.