Controlling protein association and subcellular localization with a synthetic ligand that induces heterodimerization of proteins.

Extracellular growth and differentiation factors induce changes in gene expression in the nucleus by initiating a series of protein associations that alter the subcellular localization of intracellular signaling proteins. Initial events involve receptor homo- or heterodimerization and subsequent recruitment of cytosolic signaling proteins to the inner leaflet of the plasma membrane. Intermediate events involve the translocation of proteins into the nucleus. Late events involve the recruitment of transcriptional activators to the vicinity of specific genes in the nucleus, resulting in increased gene transcription. The ability to induce signals at each of these three phases of signaling pathways is illustrated by the use of a heterodimeric chemical inducer of dimerization that causes a proximal relationship between two different target proteins.

cDNA cloning and gene mapping of a candidate human cell cycle checkpoint protein.

A family of proteins involved in cell cycle progression, DNA recombination, and the detection of DNA damage has been recently identified. One of the members of this family, human ATM, is defective in the cells of patients with ataxia telangiectasia and is involved in detection and response of cells to damaged DNA. Other members include Mei-41 (Drosophila melanogaster), Mec1p (Saccharomyces cerevisiae), and Rad3 (Schizosaccharomyces pombe), which are required for the S and G2/M checkpoints, as well as FRAP (Homo sapiens) and Torl/2p (S. cerevisiae), which are involved in a rapamycin-sensitive pathway leading to G1 cell cycle progression. We report here the cloning of a human cDNA encoding a protein with significant homology to members of this family. Three overlapping clones isolated from a Jurkat T-cell cDNA library revealed a 7.9-kb open reading frame encoding a protein that we have named FRP1 (FRAP-related protein) with 2644 amino acids and a predicted molecular mass of 301 kDa. Using fluorescence in situ hybridization and a full-length cDNA FRP1 clone, the FRP1 gene has been mapped to the chromosomal locus 3q22-q24. FRP1 is most closely related to three of the PIK-related kinase family members involved in checkpoint function--Mei-41, Mec1p, and Rad3--and as such may be the functional human counterpart of these proteins.

The basic motif-leucine zipper transcription factor Nrl can positively regulate rhodopsin gene expression.

The retinal protein Nrl belongs to a distinct subfamily of basic motif-leucine zipper DNA-binding proteins and has been shown to bind extended AP-1-like sequence elements as a homo- or heterodimer. Here, we demonstrate that Nrl can positively regulate the expression of the photoreceptor cell-specific gene rhodopsin. Electrophoretic mobility-shift analysis reveals that a protein(s) in nuclear extracts from bovine retina and the Y79 human retinoblastoma cell line binds to a conserved Nrl response element (NRE) in the upstream promoter region of the rhodopsin gene. Nrl or an antigenically similar protein is shown to be part of the bound protein complex by supershift experiments using Nrl-specific antiserum. Cotransfection studies using an Nrl-expression plasmid and a luciferase reporter gene demonstrate that interaction of the Nrl protein with the -61 to -84 region of the rhodopsin promoter (which includes the NRE) stimulates expression of the reporter gene in CV-1 monkey kidney cells. This Nrl-mediated transactivation is specifically inhibited by coexpression of a naturally occurring truncated form of Nrl (dominant negative effect). Involvement of Nrl in photoreceptor gene regulation and its continued high levels of expression in the adult retina suggest that Nrl plays a significant role in controlling retinal function.

Specificity of dimer formation in tropomyosins: influence of alternatively spliced exons on homodimer and heterodimer assembly.

Tropomyosins consist of nearly 100% alpha-helix and assemble into parallel and in-register coiled-coil dimers. In vitro it has been established that nonmuscle as well as native muscle tropomyosins can form homodimers. However, a mixture of muscle alpha and beta tropomyosin subunits results in the formation of the thermodynamically more stable alpha/beta heterodimer. Although the assembly preference of the muscle tropomyosin heterodimer can be understood thermodynamically, the presence of multiple tropomyosin isoforms expressed in nonmuscle cells points toward a more complex principle for determining dimer formation. We have investigated the dimerization of rat tropomyosins in living cells by the use of epitope tagging with a 16-aa sequence of the influenza hemagglutinin. Employing transfection and immunoprecipitation techniques, we have analyzed the dimers formed by muscle and nonmuscle tropomyosins in rat fibroblasts. We demonstrate that the information for homo- versus heterodimerization is contained within the tropomyosin molecule itself and that the information for the selectivity is conferred by the alternatively spliced exons. These results have important implications for models of the regulation of cytoskeletal dynamics.

Retinoid X receptor alpha forms tetramers in solution.

Protein-protein interactions allow the retinoid X receptor (RXR) to bind to cognate DNA as a homo- or a heterodimer and to participate in mediating the effects of a variety of hormones on gene transcription. Here we report a systematic study of the oligomeric state of RXR in the absence of a DNA template. We have used electrophoresis under nondenaturing conditions and chemical crosslinking to show that in solution, RXR alpha forms homodimers as well as homotetramers. The dissociation constants governing dimer and tetramer formation were estimated by fluorescence anisotropy studies. The results indicate that RXR tetramers are formed with a high affinity and that at protein concentrations higher than about 70 nM, tetramers will constitute the predominant species. Tetramer formation may provide an additional level of the regulation of gene transcription mediated by RXRs.

Hippocampal long-term potentiation is impaired in mice lacking brain-derived neurotrophic factor.

Brain-derived neurotrophic factor (BDNF), a member of the nerve growth factor (NGF) gene family, has been shown to influence the survival and differentiation of specific classes of neurons in vitro and in vivo. The possibility that neurotrophins are also involved in processes of neuronal plasticity has only recently begun to receive attention. To determine whether BDNF has a function in processes such as long-term potentiation (LTP), we produced a strain of mice with a deletion in the coding sequence of the BDNF gene. We then used hippocampal slices from these mice to investigate whether LTP was affected by this mutation. Homo- and heterozygous mutant mice showed significantly reduced LTP in the CA1 region of the hippocampus. The magnitude of the potentiation, as well as the percentage of cases in which LTP could be induced successfully, was clearly affected. According to the criteria tested, important pharmacological, anatomical, and morphological parameters in the hippocampus of these animals appear to be normal. These results suggest that BDNF might have a functional role in the expression of LTP in the hippocampus.

Photoconductivity and Electro-Optical Properties of Wide Band Gap Metal Oxides

The PhD activity described in this Thesis was focused on the study of metal-oxide wide-bandgap materials, aiming at fabricating new optoelectronic devices such as solar-blind UV photodetectors, high power electronics, and gas sensors. Photocurrent spectroscopy and DC photocurrent time evolution were used to investigate the performance of prototypes under different atmospheres, temperatures and excitation wavelengths (or dark conditions). Cathodoluminescence, absorption spectroscopy, XRD and SEM were used to assess structural, morphologic, electrical and optical properties of materials. This thesis is divided into two main sections, each describing the work done on a different metal-oxide semiconductor. 1) MOVPE-grown Ga2O3 thin films for UV solar-blind photodetectors and high power devices The semiconducting oxides, among them Ga2O3, have been employed for several decades as transparent conducting oxide (TCO) electrodes for fabrication of solar cells, displays, electronic, and opto-electronic devices. The interest was mainly confined to such applications, as these materials tend to grow intrinsically n-type, and attempts to get an effective p-type doping has consistently failed. The key requirements of TCO electrodes are indeed high electrical conductivity and good transparency, while crystallographic perfection is a minor issue. Furthermore, for a long period no high-quality substrates and epi-layers were available, which in turn impeded the development of a truly full-oxide electronics. Recently, Ga2O3 has attracted renewed interest, as large single crystals and high-quality homo- and hetero-epitaxial layers became available, which paved the way to novel application areas. Our research group spent the last two years in developing a low temperature (500-700°C) MOVPE growth procedure to obtain thin films of Ga2O3 on different substrates (Dept. of Physics and IMEM-CNR at UNIPR). We obtained a significant result growing on oriented sapphire epitaxial films of high crystalline, undoped, pure phase -Ga2O3 (hexagonal). The crystallographic properties of this phase were investigated by XRD, in order to clarify the lattice parameters of the hexagonal cell. First design and development of solar blind UV photodetectors based on -phase was carried out and the optoelectronic performance is evaluated by means of photocurrent spectroscopy. The UV-response is adequately fast and reliable to render this unusual phase a subject of great interest for future applications. The availability of a hexagonal phase of Ga2O3 stable up to 700°C, belonging to the same space group of gallium nitride, with high crystallinity and tunable electrical properties, is intriguing in view of the development of nitride-based devices, by taking advantage of the more favorable symmetry and epitaxial relationships with respect to the monoclinic β-phase. In addition, annealing at temperatures higher than 700°C demonstrate that the hexagonal phase converts totally in the monoclinic one. 2) ZnO nano-tetrapods: charge transport mechanisms and time-response in optoelectronic devices and sensors Size and morphology of ZnO at the nanometer scale play a key role in tailoring its physical and chemical properties. Thanks to the possibility of growing zinc oxide in a variety of different nanostructures, there is a great variety of applications, among which gas sensors, light emitting diodes, transparent conducting oxides, solar cells. Even if the operation of ZnO nanostructure-based devices has been recently demonstrated, the mechanisms of charge transport in these assembly is still under debate. The candidate performed an accurate investigation by photocurrent spectroscopy and DC-photocurrent time evolution of electrical response of both single-tetrapod and tetrapod-assembly devices. During the research done for this thesis, a thermal activation energy enables the performance of samples at high temperatures (above about 300°C). The energy barrier is related to the leg-to-leg interconnection in the assembly of nanotetrapods. Percolation mechanisms are responsible for both the very slow photo-response (minutes to hours or days) and the significant persistent photocurrent. Below the bandgap energy, electronic states were investigated but their contribution to the photocurrent are two-three order of magnitude lower than the band edge. Such devices are suitable for employ in photodetectors as well as in gas sensors, provided that the mechanism by which the photo-current is generated and gas adsorption on the surface modify the conductivity of the material are known.

Modelos de desarrollo y sistemas de gestión pública: el SNIP peruano, 2000-2014

Esta Tesis tiene dos partes. La Primera Parte es Teórica y Metodológica y trata de la actual crisis de paradigma en las Ciencias Sociales, y de cómo se puede remontar con la Teoría del Pensamiento Complejo, siempre que sus propuestas se centren en modelos empíricos de Análisis de Redes Sociales debidamente matematizados y estadísticamente refrendados. La propuesta del tesista propone enriquecer el actual homo economicus, incorporando la importancia de las relaciones con el grupo (coactivas, coercitivas o motivacionales), a través de un nuevo objeto de estudio: los Proyectos. Es mediante los Proyectos, donde los individuos y los grupos en los que interactúan, transan y organizan sus esfuerzos. El problema reside en que, no existe hasta la fecha, una sistematización y modelización de los Proyectos como objeto de estudio en las Ciencias Sociales. Sin embargo, hay una amplia experiencia de análisis y sistematización de Proyectos tanto en la Economía de la Empresa (Management, Business Administration), como en la Economía Pública. En esta Tesis se estudia todo lo publicado recientemente sobre los Proyectos de Inversión Pública (PIPs) y su eficiencia en Latinoamérica. En la Segunda Parte, centrada en un Trabajo Empírico y su modelización, el tesista crea una Base de Datos (BdD) primaria, a partir del Banco de Proyectos (BdP) del Ministerio de Economía y Finanzas (MEF) del Perú (2001-2014), que recoge todos los Proyectos de Inversión Pública (PIP), cerca de 400.000 PIPs Iniciales, los tabula en 48 categorías y posteriormente, “deja hablar a los datos” jugando a relacionar, correlacionar, inducir hipótesis y verificarlas mediante un sistema que se centra en la operativa tipo “Big Data”. A esto le denomina “triangular” porque mezcla en el esfuerzo, herramientas de Estadística Descriptiva, Estadística Inferencial y Econometría para poder refrendar el conocimiento inducido, que siempre en ciencia, es una mera certeza probabilística. El tesista concluye que en el caso del Sistema Nacional de Inversión Pública del Perú (SNIP) y más específicamente, de los procesos administrativos que emplea -denominados “Ciclo PIP”-, queda claro que se está trabajando con “fenómenos emergentes” cuyo comportamiento no se adapta a una Distribución Normal. Y que dicho comportamiento errático se debe a que la Inversión Pública es cíclica (Ecuación Evolutiva de Price) y a que el “Ciclo PIP” opera a todo nivel (GN, GR, GL) en función de las relaciones entre los miembros que componen su red. Ergo, es un tema a Analizar con Social Network Analysis (Análisis Social de Redes, ARS). El tesista concluye que las redes de “Ciclo PIP” en el Perú fallan principalmente por problemas de escasez de personal técnico multisectorial debidamente cualificado. A manera de conclusión, propone la creación de una Plataforma Web 3.0 (metadatos), que utilice un Sistema de Razonamiento Basado en Casos (SRBC) para aprovechar el conocimiento que dimana de los éxitos y fracasos de los propios PIPs, con el fin de facilitar las gestiones de los miembros de la red que formulan, evalúan y ejecutan los PIPs en el Perú, tanto a nivel Municipal (GP) como Regional (GR) y Nacional (GN).

One of Us: Joseph Conrad's Under Western Eyes and A Personal Record

Author: Torgeir Ehler Title: One of Us: Joseph Conrad's Under Western Eyes and A Personal Record Advisor: Jan Gorak Degree Date: June 2009 Abstract This present work explores the relationship of Joseph Conrad's status as a Polish exile to his creative and biographical work. Its main focus is on the tandem publications of the novel Under Western Eyes and his autobiographical volume A Personal Record, both published within a year of each other and written contemporaneously. The first chapter is a short biographical survey of Conrad's life and addresses some later biographical works by his wife, among others. An overview of critical works that deal with Under Western Eyes is presented in the second chapter. An investigation into narrative structure and its use in creating a heteroglossic text is investigated in the third chapter. How this strategy reflects Conrad's personal stake in the novel and how the novel and its creation affected the author's ability to cope with his own homo-duplex geographies is also addressed herein. The fourth chapter then concerns itself with Conrad's attempt to create a truly heteroglossic, autobiographically based persona for public consumption in Britain, while keeping true to his function as a `cultural bridge'. An early effort at communicating the exile's predicament and failure to bridge the cultural divide in the story `Amy Foster' is taken up in the fifth and final chapter. The legacy of Conrad's effort is also discussed herein as relevant to the work of Milan Kundera and Erich Maria Remarque.

Caractérisation pédologique et édaphique des sols argileux et analyse de leur occupation dans la région de Tessala (Algérie occidentale)

Les sols argileux restent dominants dans la région de Tessala (Sidi Bel Abbes) connue pour son relief érodé et accidenté où les pentes supérieures à 12% dominent. L’utilisation encore traditionnelle et irréfléchie de ces types de sols contribue à l’accélération des phénomènes d’érosion et de dessèchement agissant directement sur la végétation. Une connaissance des principales propriétés de ces sols permettra de maîtriser leurs caractéristiques physiques sources de contraintes dans leur exploitation et mise en valeur. Une maîtrise de ces éléments permettrait d’identifier des zones homo-écologiques. Il sera alors possible de travailler sur des unités physico-écologiques de mise en valeur et d’exploitation intégrée des différents types de sols. La végétation et les paramètres physiques analysés permettront de confirmer ou d’infirmer les unités de gestion préalablement déterminées. La superposition des ces zones climatique, biologique et édaphique dégagera des entités de mise en valeur claires. Une proposition d’occupation des sols selon leurs potentialités axées sur des techniques d’amélioration des caractéristiques qui constituent des entraves.

First molar size and wear within and among modern hunter-gatherers and agricultural populations

Apart from reflecting modern human dental variation, differences in dental size among populations provide a means for studying continuous evolutionary processes and their mechanisms. Dental wear, on the other hand, has been widely used to infer dietary adaptations and variability among or within diverse ancient human populations. Few such studies have focused on modern foragers and farmers, however, and diverse methods have been used. This research aimed to apply a single, standardized, and systematic quantitative procedure to measure dental size and dentin exposure in order to analyze differences among several hunter-gatherer and agricultural populations from various environments and geographic origins. In particular, we focused on sexual dimorphism and intergroup differences in the upper and lower first molars. Results indicated no sexual dimorphism in molar size and wear within the studied populations. Despite the great ethnographic variation in subsistence strategies among these populations, our findings suggest that differences in sexual division of labor do not affect dietary wear patterns.

On the forbidden gap of finite graphene nanoribbons

The electronic structure of isolated finite graphene nanoribbons is investigated by solving, at the Hartree-Fock (HF) level, the Pariser, Parr and Pople (PPP) many-body Hamiltonian. The study is mainly focused on 7-AGNR and 13-AGNR (Armchair Graphene Nano-Ribbons), whose electronic structures have been recently experimentally investigated. Only paramagnetic solutions are considered. The characteristics of the forbidden gap are studied as a function of the ribbon length. For a 7-AGNR, the gap monotonically decreases from a maximum value of ~6.5 eV for short nanoribbons to a very small value of ~0.12 eV for the longer calculated systems. Gap edges are defined by molecular orbitals that are spatially localized near the nanoribbon extremes, that is, near both zig-zag edges. On the other hand, two delocalized orbitals define a much larger gap of about 5 eV. Conductance measurements report a somewhat smaller gap of ~3 eV. The small real gap lies in the middle of the one given by extended states and has been observed by STM and reproduced by DFT calculations. On the other hand, the length dependence of the gap is not monotonous for a 13-AGNR. It decreases initially but sharply increases for lengths beyond 30 Å remaining almost constant thereafter at a value of ~2.1 eV. Two additional states localized at the nanoribbon extremes show up at energies 0.31 eV below the HOMO (Highest Occupied Molecular Orbital) and above the LUMO (Lowest Unoccupied Molecular Orbital). These numbers compare favorably with those recently obtained by means of STS for a 13-AGNR sustained by a gold surface, namely 1.4 eV for the energy gap and 0.4 eV for the position of localized band edges. We show that the important differences between 7- and 13-AGNR should be ascribed to the charge rearrangement near the zig-zag edges obtained in our calculations for ribbons longer than 30 Å, a feature that does not show up for a 7-AGNR no matter its length.

Perfeccionismo socialmente prescrito como predictor del alto rechazo a la escuela

El creciente interés en el estudio del comportamiento de rechazo a la escuela se debe a las consecuencias negativas asociadas a esta problemática. Conocer aquellas variables psicoeducativas que predicen esta conducta resulta de interés para la comunidad científica en la actualidad. El objetivo de esta investigación fue analizar la capacidad predictiva el Perfeccionismo Socialmente Prescrito sobre las altas puntuaciones en rechazo escolar. En este estudio participaron 537 estudiantes españoles entre 8 y 12 años. Los instrumentos utilizados fueron la School Refusal Assessment Scale Revised for Children (SRAS-R-C), para medir los cuatro factores que subyacen al comportamiento de rechazo a la escuela (I. Evitar la Afectividad Negativa; II. Escapar de la Aversión Social o Evaluación; III. Búsqueda de la Atención de otras Personas Significativas; IV. Búsqueda de Refuerzos Tangibles Fuera del Ámbito Escolar), y la Child and Adolescent Perfectionism Scale (CAPS), para evaluar la subescala de Perfeccionismo Socialmente Prescrito. A partir de los resultados obtenidos se confirma que la subescala de perfeccionismo evaluada actúa como un predictor positivo de altas puntuaciones en rechazo escolar para los factores II y III de la SRAS-R-C. Estos resultados deben ser valorados por psicólogos escolares y especialistas al evaluar la problemática del rechazo escolar siendo necesario proponer estrategias de prevención e intervención.

Estudio correlacional entre el afecto negativo y el rechazo escolar

El estudio del afecto y su relación con el comportamiento de rechazo a la escuela es un campo de investigación con escasos trabajos previos. El objetivo de estudio fue analizar la relación entre el Afecto Negativo y los distintos factores que justifican el rechazo escolar en estudiantes de Educación Primaria. La muestra estuvo compuesta por 476 estudiantes españoles con un rango de edad que osciló entre los 8 y los 12 años (M = 10.08; DE = 1.28). Las variables de este estudio fueron evaluadas mediante el 10-Item Positive and Negative Affect Schedule for Children (PANAS-C), para medir la subescala de Afecto Negativo, y la School Refusal Assessment Scale-Revised for Children (SRAS-R-C), para evaluar los cuatro factores que justifican el comportamiento de rechazo a la escuela (I. Evitar la Afectividad Negativa; II. Escapar de la Aversión Social o Evaluación; III. Búsqueda de la Atención de otras Personas Significativas; IV. Búsqueda de Refuerzos Tangibles Fuera del Ámbito Escolar). Los resultados revelaron que existían correlaciones positivas y significativas entre el Afecto Negativo y los tres primeros factores de la SRAS-R-C, mientras que no resultaron significativas para el cuarto factor. Los resultados obtenidos confirman la relación entre sentimientos de afectividad negativa y el rechazo escolar, aspecto que debe ser considerado en el estudio de estos casos.

AFLORAMENTO RACIONAL DO MONOTEÍSMO TRINITÁRIO

O lema sola ratione, que preside a todo o pensamento de Santo Anselmo, gera vários momentos de audácia racional na sua obra, o primeiro dos quais emerge no Monologion onde, a par de uma procura racional acerca da existência e atributos de Deus, deparamos com uma abordagem da perspectiva trinitária sem recorrer à Revelação, tal como os monges lhe tinham solicitado. Esta é, portanto, a primeira audácia da sua obra, a preceder e como que a preparar a maior delas acerca da existência de Deus no Proslogion, Liber Apologeticus e, finalmente, Cur Deus Homo. Unam quidem et unitatem propter unam essentiam, trinam vero et trinitatem propter tres nescio quid (Mon., cap. 79): à luz desta expressão de inefabilidade não poderá deixar de ler-se o que a precede e o que se lhe segue, quer nesta primeira grande obra, quer naquelas em que o tema do mistério trinitário foi tratado ou discutido.