HLA-A Typing in formalin-fixed paraffin embedded tissue samples : towards potential retrospective analysis

El Antígeno Leucocitario Humano (HLA en inglés) ha sido descrito en muchos casos como factor de pronóstico para cáncer. La característica principal de los genes de HLA, localizados en el cromosoma 6 (6p21.3), son sus numerosos polimorfismos. Los análisis de secuencia de nucleótidos muestran que la variación está restringida predominantemente a los exones que codifican los dominios de unión a péptidos de la proteína. Por lo tanto, el polimorfismo del HLA define el repertorio de péptidos que se unen a los alotipos de HLA y este hecho define la habilidad de un individuo para responder a la exposición a muchos agentes infecciosos durante su vida. La tipificación de HLA se ha convertido en un análisis importante en clínica. Muestras de tejido embebidas en parafina y fijadas con formalina (FFPE en inglés) son recolectadas rutinariamente en oncología. Este procedimiento podría ser utilizado como una buena fuente de ADN, dado que en estudios en el pasado los ensayos de recolección de ADN no eran normalmente llevados a cabo de casi ningún tejido o muestra en procedimientos clínicos regulares. Teniendo en cuenta que el problema más importante con el ADN de muestras FFPE es la fragmentación, nosotros propusimos un nuevo método para la tipificación del alelo HLA-A desde muestras FFPE basado en las secuencias del exón 2, 3 y 4. Nosotros diseñamos un juego de 12 cebadores: cuatro para el exón 2 de HLA-A, tres para el exón 3 de HLA-A y cinco para el exón 4 de HLA-A, cada uno de acuerdo las secuencias flanqueantes de su respectivo exón y la variación en la secuencia entre diferentes alelos. 17 muestran FFPE colectadas en el Hospital Universitario de Karolinska en Estocolmo Suecia fueron sometidas a PCR y los productos fueron secuenciados. Finalmente todas las secuencias obtenidas fueron analizadas y comparadas con la base de datos del IMGT-HLA. Las muestras FFPE habían sido previamente tipificadas para HLA y los resultados fueron comparados con los de este método. De acuerdo con nuestros resultados, las muestras pudieron ser correctamente secuenciadas. Con este procedimiento, podemos concluir que nuestro estudio es el primer método de tipificación basado en secuencia que permite analizar muestras viejas de ADN de las cuales no se tiene otra fuente. Este estudio abre la posibilidad de desarrollar análisis para establecer nuevas relaciones entre HLA y diferentes enfermedades como el cáncer también.

La estructura de oportunidad política en la aparición del movimiento estudiantil mexicano de 1968

Este documento presenta un estudio de cómo los cambios en las dimensiones pertenecientes a la estructura de oportunidad política, fueron determinantes en la conformación identitaria de un movimiento, la estructuración de sus demandas y sus métodos de actuación

Territorialización de la inseguridad ciudadana en Bogotá: La Calle del Bronx

La tesis devela la connotación sistemática y multicausal de lo que a través de la investigación se denomina como "Procesos de Territorialización de la Inseguridad Ciudadana". Mediante un estudio de caso, se pone en evidencia la apropiación y captura sostenida en el tiempo de fenómenos como la inseguridad y la criminalidad, sobre determinadas zonas o barrios urbanos que por sus características socioeconómicas, políticas, geográficas, culturales, laborales y de mercado de quienes los habitan o frecuentan, se consideran como sectores "críticos y/o vulnerables".

Los intereses estratégicos de EEUU, Rusia, China y Japón como determinantes de la estrategia de mantenimiento de la Estructura en la Península Coreana. 2003-2013.

El propósito principal de esta monografía es ofrecer una perspectiva crítica sobre el conflicto latente en la Península Coreana, haciendo un acercamiento al mismo desde un marco teórico asentado en el realismo estructural de Kenneth Waltz. De este modo, se busca responder a cuestiones sobre los intereses estatales como fundamento básico de las estrategias de mantenimiento de la Estructura en regiones geopolíticamente sensibles. Al final, se llega a la conclusión afirmando que la Estructura ejerce una serie de funciones para garantizar su preservación mediante una acción de convergencia en la conducta de los Estados. Esta realidad ha mantenido a la Península Coreana sin un conflicto bélico en los últimos 50 años, muy a pesar de estar al borde del mismo en varias ocasiones, ya que de llegarse a presentar se rompería la estabilidad de la región, y por ende el Equilibrio de Poderes estaría en grave riesgo.

Desequilibrio entre lenguas y rendimiento escolar : el lenguaje como mediador del éxito académico en contextos bilingües.

Analizar el nivel de éxito-fracaso escolar entre alumnos de Aragón y de Asturias que reciben enseñanza exclusivamente en lengua castellana y alumnos a los que se les imparten clases optativas en catalán y en asturiano. En el caso aragonés, los objetivos son: evaluar el conocimiento matemático desarrollado por los escolares catalanoparlantes de Aragón, tomando como referencia el correspondiente a sus coetáneos que habitan en territorios aragoneses monolingües; establecer, en el caso de que hubiese diferencias, los factores (individuales, sociales o del currículum) explicativos de los mismos; orientar, en función de los análisis precedentes y ante perspectivas de cambios a corto plazo, derivados del traspaso de competencias educativas y la implantación de la LOGSE, hacia líneas de actuación educativa más acordes con la naturaleza social bilingüe de la Franja aragonesa. Sus hipótesis son: 1. El alumnado catalanoparlante de Aragón que asiste a clases de catalán, evidencia un conocimiento matemático superior al de sus coetáneos que no asisten a dichas clases. 2. Los escolares del grupo control obtienen resultado aproximados, en cuanto a conocimiento matemático, que los escolares catanoparlantes de Aragón que asisten a clases de lengua catalana. 3. Los escolares del grupo control obtienen resultados superiores, en cuanto a conocimiento matemático, que los escolares catalanoparlantes de Aragón que no asisten a clases de lengua catalana. En el caso asturiano, las hipótesis son: 1. El alumnado asturiano que asiste a clases de lengua asturiana, evidencia un conocimiento de lengua castellana superior al de sus coetáneos que no asisten a dichas clases. 2. El alumnado asturiano que asiste a clases de lengua asturiana, evidencia un conocimiento matemático superior al de sus coetáneos que no asisten a dichas clases. 3. Existirá una correlación positiva entre el conocimeinto lingüístico en castellano y el conocimiento matemático del alumnado. 240 alumnos y alumnas de centros públicos de enseñanza primaria y de segundo de ESO de municipios de la comarca del Bajo Cinca, en el caso aragonés; 231 alumnos y alumnas de 6 centros de enseñanza primaria que imparten el primer ciclo de Secundaria y 8 Institutos de Enseñanza Secundaria, en el caso asturiano. Las variables estudiadas en el caso aragonés fueron: condición lingüística familiar (CLF), opcionalidad (OPC), situación socioprofesional (SSP), cociente intelectual (CI), sector y conocimiento matemático (IAEP); en el caso asturiano, las variables fueron: condición lingüística familiar (CLF), situación socioprofesional (SSP), opcionalidad (OPC), cociente intelectual (CI) y centro. Entre otros, los instrumentos utilizados fueron la evaluación del conocimiento matemático elaborada por la IAEP, y en el caso asturiano además, la evaluación de conocimiento lingüístico castellano. Statview 1.04 y técnicas descriptivas. Los resultados finales muestran que los escolares catalanoparlantes de Aragón que asisten a clases de lengua catalana, al estructurar su propia lengua, obtienen un beneficio añadido en conocimiento de lengua castellana que incide directamente en un mejor rendimiento matemático comparado con aquellos que no asisten a dichas clases. Además, no aparecen diferencias significativas con sus coetáneos que habitan territorios aragoneses monolingües, mientras que sí aparecen entre éstos últimos y los escolares catalanoparlantes que no asisten a clases de lengua catalana. Respecto a la muestra asturiana, los datos ponen de manifiesto una mejora en el conocimiento lingüístico castellano en aquel alumnado que asiste a clases de asturiano. Se observa también una correlación positiva entre conocimiento lingüístico castellano y conocimiento matemático. En cuanto al conocimiento matemático, aparecen diferencias notables entre el alumnado que cursa sus estudios en centros de Primaria y centros de Secundaria, lo que se interpreta en función del pobre estatus del asturiano y la poca tradición en su incorporación como materia del currículum en los IES.

Global transcript analysis of rice leaf and seed using SAGE technology.

We have compiled two comprehensive gene expression profiles from mature leaf and immature seed tissue of rice (Oryza sativa ssp. japonica cultivar Nipponbare) using Serial Analysis of Gene Expression (SAGE) technology. Analysis revealed a total of 50 519 SAGE tags, corresponding to 15 131 unique transcripts. Of these, the large majority (approximately 70%) occur only once in both libraries. Unexpectedly, the most abundant transcript (approximately 3% of the total) in the leaf library was derived from a type 3 metallothionein gene. The overall frequency profiles of the abundant tag species from both tissues differ greatly and reveal seed tissue as exhibiting a non-typical pattern of gene expression characterized by an over abundance of a small number of transcripts coding for storage proteins. A high proportion ( approximately 80%) of the abundant tags (> or = 9) matched entries in our reference rice EST database, with many fewer matches for low abundant tags. Singleton transcripts that are common to both tissues were collated to generate a summary of low abundant transcripts that are expressed constitutively in rice tissues. Finally and most surprisingly, a significant number of tags were found to code for antisense transcripts, a finding that suggests a novel mechanism of gene regulation, and may have implications for the use of antisense constructs in transgenic technology.

Biological control of black vine weevil (Otiorhynchus sulcatus, Coleoptera: Curculionidae) by entomopathogenic bacteria and their cell-free toxic metabolites

Biocontrol agents such as Xeiwrhabduf, nemalophilci and X. nematophila ssp. bovienii and their cell-free protein toxin complexes were lethal to larvae of O. sulcatus when applied to potting compost in the absence of plants. Similarly, strawberry plants infected with 0. sulcaitfi larvae were protected from damage by applications of both cell suspensions of the bacteria and solutions of their cell-free toxic metabolites, indicating that it is the protein toxins, which are responsible for the lethal effects observed. These toxic metabolites were found more effective against 0. sulccitus larvae when treated in soil microflora. Insect mortality is increased by increasing temperature and bacterial concentration. The toxins remained pathogenic for several months when stored in potting soil either at 15 or 20°C, however, bacterial cells were not as persistent as the toxins. It is therefore suggested that these bacteria and their toxic metabolites can he applied in soil for insect pest control.

Mapping the landscape of the lymphocytic choriomeningitis virus stable signal peptide reveals novel functional domains

The stable signal peptide (SSP) of the lymphocytic choriomeningitis virus surface glycoprotein precursor has several unique characteristics. The SSP is unusually long, at 58 amino acids, and contains two hydrophobic domains, and its sequence is highly conserved among both Old and New World arenaviruses. To better understand the functions of the SSP, a panel of point and deletion mutants was created by in vitro mutagenesis to target the highly conserved elements within the SSP. We were also able to confirm critical residues required for separate SSP functions by trans-complementation. Using these approaches, it was possible to resolve functional domains of the SSP. In characterizing our SSP mutants, we discovered that the SSP is involved in several distinct functions within the viral life cycle, beyond translocation of the viral surface glycoprotein precursor into the endoplasmic reticulum lumen. The SSP is required for efficient glycoprotein expression, posttranslational maturation cleavage of GP1 and GP2 by SKI-1/S1P protease, glycoprotein transport to the cell surface plasma membrane, formation of infectious virus particles, and acid pH-dependent glycoprotein-mediated cell fusion.

Grey-box radial basis function modelling: the art of incorporating prior knowledge

A basic principle in data modelling is to incorporate available a priori information regarding the underlying data generating mechanism into the modelling process. We adopt this principle and consider grey-box radial basis function (RBF) modelling capable of incorporating prior knowledge. Specifically, we show how to explicitly incorporate the two types of prior knowledge: the underlying data generating mechanism exhibits known symmetric property and the underlying process obeys a set of given boundary value constraints. The class of orthogonal least squares regression algorithms can readily be applied to construct parsimonious grey-box RBF models with enhanced generalisation capability.

Subspace-based blind adaptive multiuser detection for multirate DS/CDMA signals

The existing dual-rate blind linear detectors, which operate at either the low-rate (LR) or the high-rate (HR) mode, are not strictly blind at the HR mode and lack theoretical analysis. This paper proposes the subspace-based LR and HR blind linear detectors, i.e., bad decorrelating detectors (BDD) and blind MMSE detectors (BMMSED), for synchronous DS/CDMA systems. To detect an LR data bit at the HR mode, an effective weighting strategy is proposed. The theoretical analyses on the performance of the proposed detectors are carried out. It has been proved that the bit-error-rate of the LR-BDD is superior to that of the HR-BDD and the near-far resistance of the LR blind linear detectors outperforms that of its HR counterparts. The extension to asynchronous systems is also described. Simulation results show that the adaptive dual-rate BMMSED outperform the corresponding non-blind dual-rate decorrelators proposed by Saquib, Yates and Mandayam (see Wireless Personal Communications, vol. 9, p.197-216, 1998).

Flexible MMSE DFE using length and LAG adaptation

This paper investigates how to choose the optimum tap-length and decision delay for the decision feedback equalizer (DFE). Although the feedback filter length can be set as the channel memory, there is no closed-form expression for the feedforward filter length and decision delay. In this paper, first we analytically show that the two dimensional search for the optimum feedforward filter length and decision delay can be simplified to a one dimensional search, and then describe a new adaptive DFE where the optimum structural parameters can be self-adapted.

Inner plasma structure of the low-latitude reconnection layer

We report a clear transition through a reconnection layer at the low-latitude magnetopause which shows a complete traversal across all reconnected field lines during northwestward interplanetary magnetic field (IMF) conditions. The associated plasma populations confirm details of the electron and ion mixing and the time history and acceleration through the current layer. This case has low magnetic shear with a strong guide field and the reconnection layer contains a single density depletion layer on the magnetosheath side which we suggest results from nearly field-aligned magnetosheath flows. Within the reconnection boundary layer, there are two plasma boundaries, close to the inferred separatrices on the magnetosphere and magnetosheath sides (Ssp and Ssh) and two boundaries associated with the Alfvén waves (or Rotational Discontinuities, RDsp and RDsh). The data are consistent with these being launched from the reconnection site and the plasma distributions are well ordered and suggestive of the time elapsed since reconnection of the field lines observed. In each sub-layer between the boundaries the plasma distribution is different and is centered around the current sheet, responsible for magnetosheath acceleration. We show evidence for a velocity dispersion effect in the electron anisotropy that is consistent with the time elapsed since reconnection. In addition, new evidence is presented for the occurrence of partial reflection of magnetosheath electrons at the magnetopause current layer.

Genetic structure and linkage disequilibrium in landrace populations of barley in Sardinia

Multilocus digenic linkage disequilibria (LD) and their population structure were investigated in eleven landrace populations of barley (Hordeum vulgare ssp. vulgare L.) in Sardinia, using 134 dominant simple-sequence amplified polymorphism markers. The analysis of molecular variance for these markers indicated that the populations were partially differentiated (F ST = 0.18), and clustered into three geographic areas. Consistent with this population pattern, STRUCTURE analysis allocated individuals from a bulk of all populations into four genetic groups, and these groups also showed geographic patterns. In agreement with other molecular studies in barley, the general level of LD was low (13 % of locus pairs, with P < 0.01) in the bulk of 337 lines, and decayed steeply with map distance between markers. The partitioning of multilocus associations into various components indicated that genetic drift and founder effects played a major role in determining the overall genetic makeup of the diversity in these landrace populations, but that epistatic homogenising or diversifying selection was also present. Notably, the variance of the disequilibrium component was relatively high, which implies caution in the pooling of barley lines for association studies. Finally, we compared the analyses of multilocus structure in barley landrace populations with parallel analyses in both composite crosses of barley on the one hand and in natural populations of wild barley on the other. Neither of these serves as suitable mimics of landraces in barley, which require their own study. Overall, the results suggest that these populations can be exploited for LD mapping if population structure is controlled.

PCR-based markers diagnostic for spring and winter seasonal growth habit in barley

Toward the ultimate goal of replacing field-based evaluation of seasonal growth habit, we describe the design and validation of a multiplex polymerase chain reaction assay diagnostic for allelic status at the barley (Hordeum vulgare ssp. vulgare L.) vernalization locus, VRN-H1 By assaying for the presence of all known insertion–deletion polymorphisms thought to be responsible for the difference between spring and winter alleles, this assay directly tests for the presence of functional polymorphism at VRN-H1 Four of the nine previously recognized VRN-H1 haplotypes (including both winter alleles) give unique profiles using this assay. The remaining five spring haplotypes share a single profile, indicative of function-altering deletions spanning, or adjacent to, the putative “vernalization critical” region of intron 1. When used in conjunction with a previously published PCR-based assay diagnostic for alleles at VRN-H2, it was possible to predict growth habit in all the 100 contemporary UK spring and winter lines analyzed in this study. This assay is likely to find application in instances when seasonal growth habit needs to be determined without the time and cost of phenotypic assessment and during marker-assisted selection using conventional and multicross population analysis.

Distribution of calcium (Ca) and magnesium (Mg) in the leaves of Brassica rapa under varying exogenous Ca and Mg supply

Background and Aims Leafy vegetable Brassica crops are an important source of dietary calcium (Ca) and magnesium (Mg) and represent potential targets for increasing leaf Ca and Mg concentrations through agronomy or breeding. Although the internal distribution of Ca and Mg within leaves affects the accumulation of these elements, such data are not available for Brassica. The aim of this study was to characterize the internal distribution of Ca and Mg in the leaves of a vegetable Brassica and to determine the effects of altered exogenous Ca and Mg supply on this distribution. Methods Brassica rapa ssp. trilocularis ‘R-o-18’ was grown at four different Ca:Mg treatments for 21 d in a controlled environment. Concentrations of Ca and Mg were determined in fully expanded leaves using inductively coupled plasma-mass spectrometry (ICP-MS). Internal distributions of Ca and Mg were determined in transverse leaf sections at the base and apex of leaves using energy-dispersive X-ray spectroscopy (EDS) with cryo-scanning electron microscopy (cryo-SEM). Key Results Leaf Ca and Mg concentrations were greatest in palisade and spongy mesophyll cells, respectively, although this was dependent on exogenous supply. Calcium accumulation in palisade mesophyll cells was enhanced slightly under high Mg supply; in contrast, Mg accumulation in spongy mesophyll cells was not affected by Ca supply. Conclusions The results are consistent with Arabidopsis thaliana and other Brassicaceae, providing phenotypic evidence that conserved mechanisms regulate leaf Ca and Mg distribution at a cellular scale. The future study of Arabidopsis gene orthologues in mutants of this reference B. rapa genotype will improve our understanding of Ca and Mg homeostasis in plants and may provide a model-to-crop translation pathway for targeted breeding.