Genetic association study of QT interval highlights role for calcium signaling pathways in myocardial repolarization

The QT interval, an electrocardiographic measure reflecting myocardial repolarization, is a heritable trait. QT prolongation is a risk factor for ventricular arrhythmias and sudden cardiac death (SCD) and could indicate the presence of the potentially lethal mendelian long-QT syndrome (LQTS). Using a genome-wide association and replication study in up to 100,000 individuals, we identified 35 common variant loci associated with QT interval that collectively explain ∼8-10% of QT-interval variation and highlight the importance of calcium regulation in myocardial repolarization. Rare variant analysis of 6 new QT interval-associated loci in 298 unrelated probands with LQTS identified coding variants not found in controls but of uncertain causality and therefore requiring validation. Several newly identified loci encode proteins that physically interact with other recognized repolarization proteins. Our integration of common variant association, expression and orthogonal protein-protein interaction screens provides new insights into cardiac electrophysiology and identifies new candidate genes for ventricular arrhythmias, LQTS and SCD.

REV7 counteracts DNA double-strand break resection and affects PARP inhibition

Error-free repair of DNA double-strand breaks (DSBs) is achieved by homologous recombination (HR), and BRCA1 is an important factor for this repair pathway. In the absence of BRCA1-mediated HR, the administration of PARP inhibitors induces synthetic lethality of tumour cells of patients with breast or ovarian cancers. Despite the benefit of this tailored therapy, drug resistance can occur by HR restoration. Genetic reversion of BRCA1-inactivating mutations can be the underlying mechanism of drug resistance, but this does not explain resistance in all cases. In particular, little is known about BRCA1-independent restoration of HR. Here we show that loss of REV7 (also known as MAD2L2) in mouse and human cell lines re-establishes CTIP-dependent end resection of DSBs in BRCA1-deficient cells, leading to HR restoration and PARP inhibitor resistance, which is reversed by ATM kinase inhibition. REV7 is recruited to DSBs in a manner dependent on the H2AX-MDC1-RNF8-RNF168-53BP1 chromatin pathway, and seems to block HR and promote end joining in addition to its regulatory role in DNA damage tolerance. Finally, we establish that REV7 blocks DSB resection to promote non-homologous end-joining during immunoglobulin class switch recombination. Our results reveal an unexpected crucial function of REV7 downstream of 53BP1 in coordinating pathological DSB repair pathway choices in BRCA1-deficient cells.

El problema isostático inverso de Vening Meinesz. Teoría y desarrollo. Aplicación práctica para la determinación de la profundidad de la discontinuidad de Mohorovičić en la Península Ibérica

La discontinuidad de Mohorovičić, más conocida simplemente como “Moho” constituye la superficie de separación entre los materiales rocosos menos densos de la corteza y los materiales rocosos más densos del manto, suponiendo estas capas de densidad constante del orden de 2.67 y 3.27 g/cm3, y es un contorno básico para cualquier estudio geofísico de la corteza terrestre. Los estudios sísmicos y gravimétricos realizados demuestran que la profundidad del Moho es del orden de 30-40 km por debajo de la Península Ibérica y 5-15 km bajo las zonas marinas. Además las distintas técnicas existentes muestran gran correlación en los resultados. Haciendo la suposición de que el campo de gravedad de la Península Ibérica (como le ocurre al 90% de la Tierra) está isostáticamente compensado por la variable profundidad del Moho, suponiendo un contraste de densidad constante entre la corteza y el manto y siguiendo el modelo isostático de Vening Meinesz (1931), se formula el problema isostático inverso para obtener tal profundidad a partir de la anomalía Bouguer de la gravedad calculada gracias a la gravedad observada en la superficie terrestre. La particularidad de este modelo es la compensación isostática regional de la que parte la teoría, que se asemeja a la realidad en mayor medida que otros modelos existentes, como el de Airy-Heiskanen, que ha sido históricamente el más utilizado en trabajos semejantes. Además, su solución está relacionada con el campo de gravedad global para toda la Tierra, por lo que los actuales modelos gravitacionales, la mayoría derivados de observaciones satelitales, deberían ser importantes fuentes de información para nuestra solución. El objetivo de esta tesis es el estudio con detalle de este método, desarrollado por Helmut Moritz en 1990, que desde entonces ha tenido poca evolución y seguidores y que nunca se ha puesto en práctica en la Península Ibérica. Después de tratar su teoría, desarrollo y aspectos computacionales, se está en posición de obtener un modelo digital del Moho para esta zona a fin de poder utilizarse para el estudio de la distribución de masas bajo la superficie terrestre. A partir de los datos del Moho obtenidos por métodos alternativos se hará una comparación. La precisión de ninguno de estos métodos es extremadamente alta (+5 km aproximadamente). No obstante, en aquellas zonas donde exista una discrepancia de datos significaría un área descompensada, con posibles movimientos tectónicos o alto grado de riesgo sísmico, lo que le da a este estudio un valor añadido. ABSTRACT The Mohorovičić discontinuity, simply known as “Moho” constitutes the division between the rocky and less thick materials of the mantle and the heavier ones in the crust, assuming densities of the orders of 2.67 y 3.27 g/cm3 respectively. It is also a basic contour for every geophysical kind of studies about the terrestrial crust. The seismic and previous gravimetric observations done in the study area show that the Moho depth is of the order of 30-40 km beneath the ground and 5-15 km under the ocean basin. Besides, the different techniques show a good correlation in their results. Assuming that the Iberian Peninsula gravity field (as it happens for the 90% of the Earth) is isostatically compensated according to the variable Moho depth, supposing a constant density contrast between crust and mantle, and following the isostatic Vening Meinesz model (1931), the inverse isostatic problem can be formulated from Bouguer gravity anomaly data obtained thanks to the observed gravity at the surface of the Earth. The main difference between this model and other existing ones, such as Airy- Heiskanen’s (pure local compensation and mostly used in these kinds of works) is the approaching to a regional isostatic compensation, much more in accordance with reality. Besides, its solution is related to the global gravity field, and the current gravitational models -mostly satellite derived- should be important data sources in such solution. The aim of this thesis is to study with detail this method, developed by Helmut Moritz in 1990, which hardly ever has it put into practice. Moreover, it has never been used in Iberia. After studying its theory, development and computational aspects, we are able to get a Digital Moho Model of the Iberian Peninsula, in order to study the masses distribution beneath the Earth’s surface. With the depth Moho information obtained from alternative methods, a comparison will be done. Both methods give results with the same order of accuracy, which is not quite high (+ 5 km approximately). Nevertheless, the areas in which a higher difference is observed would mean a disturbance of the compensation, which could show an unbalanced area with possible tectonic movements or potential seismic risk. It will give us an important additive value, which could be used in, at first, non related fields, such as density discrepancies or natural disasters contingency plans.

Dose-response resistance to HIV-1/MuLV pseudotype virus ex vivo in a hairpin ribozyme transgenic mouse model

We have investigated the efficacy of a hairpin ribozyme targeting the 5′ leader sequence of HIV-1 RNA in a transgenic model system. Primary spleen cells derived from transgenic or control mice were infected with HIV-1/MuLV pseudotype virus. A significantly reduced susceptibility to infection in ribozyme-expressing transgenic spleen cells (P = 0.01) was shown. Variation of transgene-expression levels between littermates revealed a dose response between ribozyme expression and viral resistance, with an estimated cut off value below 0.2 copies of hairpin ribozyme per cell. These findings open up possibilities for studies on ribozyme efficacy and anti-HIV-1 gene therapy.

Activation of Androgen Receptor Function by a Novel Nuclear Protein Kinase

Androgen receptor (AR) belongs to the nuclear receptor superfamily and mediates the biological actions of male sex steroids. In this work, we have characterized a novel 130-kDa Ser/Thr protein kinase ANPK that interacts with the zinc finger region of AR in vivo and in vitro. The catalytic kinase domain of ANPK shares considerable sequence similarity with the minibrain gene product, a protein kinase suggested to contribute to learning defects associated with Down syndrome. However, the rest of ANPK sequence, including the AR-interacting interface, exhibits no apparent homology with other proteins. ANPK is a nuclear protein that is widely expressed in mammalian tissues. Its overexpression enhances AR-dependent transcription in various cell lines. In addition to the zinc finger region, ligand-binding domain and activation function AF1 of AR are needed, as the activity of AR mutants devoid of these domains was not influenced by ANPK. The receptor protein does not appear to be a substrate for ANPK in vitro, and overexpression of ANPK does not increase the extent of AR phosphorylation in vivo. In view of this, it is likely that ANPK-mediated activation of AR function is exerted through modification of AR-associated proteins, such as coregulatory factors, and/or through stabilization of the receptor protein against degradation.

Inherited susceptibility to uterine leiomyomas and renal cell cancer

Herein we report the clinical, histopathological, and molecular features of a cancer syndrome with predisposition to uterine leiomyomas and papillary renal cell carcinoma. The studied kindred included 11 family members with uterine leiomyomas and two with uterine leiomyosarcoma. Seven individuals had a history of cutaneous nodules, two of which were confirmed to be cutaneous leiomyomatosis. The four kidney cancer cases occurred in young (33- to 48-year-old) females and displayed a unique natural history. All these kidney cancers displayed a distinct papillary histology and presented as unilateral solitary lesions that had metastasized at the time of diagnosis. Genetic-marker analysis mapped the predisposition gene to chromosome 1q. Losses of the normal chromosome 1q were observed in tumors that had occurred in the kindred, including a uterine leiomyoma. Moreover, the observed histological features were used as a tool to diagnose a second kindred displaying the phenotype. We have shown that predisposition to uterine leiomyomas and papillary renal cell cancer can be inherited dominantly through the hereditary leiomyomatosis and renal cell cancer (HLRCC) gene. The HLRCC gene maps to chromosome 1q and is likely to be a tumor suppressor. Clinical, histopathological, and molecular tools are now available for accurate detection and diagnosis of this cancer syndrome.

Comprehensive copy number and gene expression profiling of the 17q23 amplicon in human breast cancer

The biological significance of DNA amplification in cancer is thought to be due to the selection of increased expression of a single or few important genes. However, systematic surveys of the copy number and expression of all genes within an amplified region of the genome have not been performed. Here we have used a combination of molecular, genomic, and microarray technologies to identify target genes for 17q23, a common region of amplification in breast cancers with poor prognosis. Construction of a 4-Mb genomic contig made it possible to define two common regions of amplification in breast cancer cell lines. Analysis of 184 primary breast tumors by fluorescence in situ hybridization on tissue microarrays validated these results with the highest amplification frequency (12.5%) observed for the distal region. Based on GeneMap'99 information, 17 known genes and 26 expressed sequence tags were localized to the contig. Analysis of genomic sequence identified 77 additional transcripts. A comprehensive analysis of expression levels of these transcripts in six breast cancer cell lines was carried out by using complementary DNA microarrays. The expression patterns varied from one cell line to another, and several overexpressed genes were identified. Of these, RPS6KB1, MUL, APPBP2, and TRAP240 as well as one uncharacterized expressed sequence tag were located in the two common amplified regions. In summary, comprehensive analysis of the 17q23 amplicon revealed a limited number of highly expressed genes that may contribute to the more aggressive clinical course observed in breast cancer patients with 17q23-amplified tumors.

Percepción de los trabajadores acerca de la seguridad en el trabajo en una empresa del sector de gas natural en Casanare

Introducción: La percepción puede ser considerada principalmente como un grupo de procesos internos en las personas que genera en los observadores una representación mental del entorno. Percibir es adelantarse al suceso, a lo que puede llegar a suceder, determinando lo que puede ser un peligro a nuestra seguridad y salud. Objetivo: El presente estudio tiene como objetivo determinar la percepción de los trabajadores acerca de la seguridad en el trabajo en áreas de procesos automatizados y mecánicas en una empresa del sector de gas natural en el departamento de Casanare, Colombia 2016. Metodología: Se llevó a cabo un estudio de corte transversal en 50 trabajadores de una empresa del sector de gas natural en Casanare. Se uttilizó el Cuestionario nórdico NOSACQ-50-Spanish, instrumento validado para evaluar la percepción acerca de la seguridad y salud en el lugar de trabajo. Se incluyeron variables sociodemográficas, laborales y las relacionadas con la percepción de la seguridad en el lugar del trabajo. Para el análsiis estadistico se calcularon medidas de tendencia central y de dispersión. El estudio de la percepción de seguridad tuvo como puntos de corte: < 2,5 mala percepción y > 2,5 buena percepción. Se emplearon pruebas de asociación X2 o test exacto de Fisher (valores esperados <5) y medidas de asociación OR con sus intervalos de confianza del 95% y se usaron modelos de regresión lineal. Resultados: El total de trabajadores fue de 50 personas, el 76% correspondió al sexo masculino y la mayor distribución se presentó en áreas mecánicas con 52% frente al 48% que se encontró en áreas automatizadas. Se halló asociación estadísticamente significativa entre las variables sociodemográficas área/sexo (Pr = 0,016), indicando que a los trabajadores de sexo femenino se les ubican en el área mecánica mientras que los de sexo masculino, laboran en todas las áreas sin darle relevancia al género. También se encontró asociación con las variables área/año de nacimiento (Pr =0.022), indicando que en el área automatizada se dejan profesionales con promedio de edad de 32 años, por el tipo de requisitos en competencias que exigen para ejecutar las labores. Para las demás variables ocupacionales que evaluaron la percepción de seguridad frente al trabajo, no se encontró asociación significativa. Conclusión: Los resultados permiten realizar acciones en pro de mejorar la percepción de los trabajadores dentro de la organización. Se podrán desarrollar programas de seguridad y salud en el trabajo, que respondan de manera efectiva a los peligros laborales detectados.