Modelo CCa, plantilla de Word para el montaje de documentos de tesis

El Centro de Investigaciones y Referencias de Aterosclerosis de la Habana desarrolla la Carrera Certificativa de Investigaciones en Aterosclerosis que incluye: Diplomado, Maestría y Doctorado. La elaboración de documentos de tesis presente, ha sido un problema para los implicados en el proceso. Propósito: Crear una plantilla de Word que sirva de guía a maestrantes y doctorantes en la elaboración de sus documentos de tesis. Método: se realizó trabajo búsqueda de lo legislado sobre la creación de los documentos de tesis, se investigaron las deficiencias ya presentadas, se elaboró la plantilla de Word y un manual, se distribuyó para su validación. Resultados: Se creó el “Modelo CCa”, plantilla de Word totalmente modificable que contiene la estructura de la tesis e incluye las orientaciones metodológicas en el mismo lugar donde se inserta el contenido. Se publicó en el sitio web del centro. Conclusiones: La plantilla facilitó el desarrollo del proceso educativo, ayudó al perfeccionamiento científico de los alumnos y creó condiciones para un uso más práctico de la tecnología.</p

Sitio web CIRAH. Nuevo espacio de trabajo para el centro

Introducción: el CIRAH es una institución de postgrado que contribuye a la actualización permanente del personal de la salud en sus conocimientos sobre la enfermedad aterosclerótica, sus factores de riesgo y sus consecuencias orgánicas. Desarrolla la Carrera Certificativa de Investigación en Aterosclerosis que oferta a todo el país dividida en tres etapas: Diplomado, Maestría y Doctorado. Coordinar este proceso es complejo y el centro cuenta con muy poco personal para ello. Objetivo: crear un sitio web para el CIRAH que facilite la gestión de información y el proceso docente-investigativo de la Carrera Certificativa de Investigaciones de Aterosclerosis. Material y Método: se comenzó por realizar un trabajo de mesa para definir los contenidos del sitio analizando la misión y visión de la institución. Se revisaron sitios web temáticos, de centros de investigación y educacionales. Se seleccionó la información del centro, se hizo el diseño estructural y gráfico del sitio, se definió la plataforma a utilizar y las herramientas a emplear. Por último, se montaron todos los contenidos. Resultados: se publicó el sitio institucional CIRAH. Mostró la estructura, contenidos y resultados del trabajo desarrollado. Se colocaron variadas secciones para facilitar el acceso a la información bibliográfica. Ofreció tutoriales y herramientas útiles para el trabajo. Respondió a las dos líneas fundamentales de trabajo: docencia-investigación sobre aterosclerosis. Centralizó el acceso a la información del centro. Conclusiones: el sitio web CIRAH resultó una nueva vía de organización y gestión de información para el centro. Su lanzamiento inició una nueva forma de trabajo colaborativo para sus trabajadores. Palabras clave: CIRAH, aterosclerosis, sitio web temático, sitio web educativo, sitio institucional, sitio web de especialidad. </p

Upregulation of ASCL1 and inhibition of Notch signaling pathway characterize progressive astrocytoma

Astrocytoma is the most common type of brain cancer constituting more than half of all brain tumors. With an aim to identify markers describing astrocytoma progression, we have carried out microarray analysis of astrocytoma samples of different grades using cDNA microarray containing 1152 cancer-specific genes. Data analysis identified several differentially regulated genes between normal brain tissue and astrocytoma as well as between grades II/III astrocytoma and glioblastoma multiforme (GBM; grade IV). We found several genes known to be involved in malignancy including Achaete-scute complex-like 1 (Drosophila) (ASCL1; Hash 1). As ASCL has been implicated in neuroendocrine, medullary thyroid and small-cell lung cancers, we chose to examine the role of ASCL1 in the astrocytoma development. Our data revealed that ASCL1 is overexpressed in progressive astrocytoma as evidenced by increased levels of ASCL1 transcripts in 85.71% (6/7) of grade II diffuse astrocytoma (DA), 90% (9/10) of grade III anaplastic astrocytoma (AA) and 87.5% (7/8) of secondary GBMs, while the majority of primary de novo GBMs expressed similar to or less than normal brain levels (66.67%; 8/12). ASCL1 upregulation in progressive astrocytoma is accompanied by inhibition of Notch signaling as seen by uninduced levels of HES1, a transcriptional target of Notch1, increased levels of HES6, a dominant-negative inhibitor of HES1-mediated repression of ASCL1, and increased levels of Notch ligand Delta1, which is capable of inhibiting Notch signaling by forming intracellular Notch ligand autonomous complexes. Our results imply that inhibition of Notch signaling may be an important early event in the development of grade II DA and subsequent progression to grade III AA and secondary GBM. Furthermore, ASCL1 appears to be a putative marker to distinguish primary GBM from secondary GBM

Novel glioblastoma markers with diagnostic and prognostic value identified through transcriptome analysis

Purpose: Current methods of classification of astrocytoma based on histopathologic methods are often subjective and less accurate. Although patients with glioblastoma have grave prognosis, significant variability in patient outcome is observed. Therefore, the aim of this study was to identify glioblastoma diagnostic and prognostic markers through microarray analysis. Experimental Design: We carried out transcriptome analysis of 25 diffusely infiltrating astrocytoma samples [WHO grade II - diffuse astrocytoma, grade III - anaplastic astrocytoma, and grade IV - glioblastoma (GBM)] using cDNA microarrays containing 18,981 genes. Several of the markers identified were also validated by real-time reverse transcription quantitative PCR and immunohistochemical analysis on an independent set of tumor samples (n = 100). Survival analysis was carried out for two markers on another independent set of retrospective cases (n = 51). Results: We identified several differentially regulated grade-specific genes. Independent validation by real-time reverse transcription quantitative PCR analysis found growth arrest and DNA-damage-inducible α (GADD45α) and follistatin-like 1 (FSTL1) to be up-regulated in most GBMs (both primary and secondary), whereas superoxide dismutase 2 and adipocyte enhancer binding protein 1 were up-regulated in the majority of primary GBM. Further, identification of the grade-specific expression of GADD45α and FSTL1 by immunohistochemical staining reinforced our findings. Analysis of retrospective GBM cases with known survival data revealed that cytoplasmic overexpression of GADD45α conferred better survival while the coexpression of FSTL1 with p53 was associated with poor survival. Conclusions: Our study reveals that GADD45α and FSTLI are GBM-specific whereas superoxide dismutase 2 and adipocyte enhancer binding protein 1 are primary GBM-specific diagnostic markers. Whereas GADD45α overexpression confers a favorable prognosis, FSTL1 overexpression is a hallmark of poor prognosis in GBM patients

Identification de gènes importants pour la spermatogenèse (une approche de priorisation de gènes inter-espèces)

Recent advances in genomics and high-throughput technologies have significantly contributed to our understanding of spermatogenesis by identifying numerous genes expressed during the development of male gametes. These approaches do not, however, pinpoint the essential genes and several strategies can be adopted for the selection of candidate genes for functional genomic studies. The first part of this thesis describes a screen based on conserved and differential expression in the testis from three mammalian species. Homologs of candidate genes were inactivated in the C. elegans model and worms were assayed for infertility phenotypes. The second part describes the construction of a tool for 'prioritizing' candidate genes based on phenotype and other data of relevance to the subject. We implemented a system capable of ranking genes for spermatogenesis based on multiple types of genomic information. The resulting Gene Prioritization System (GPSy) is currently the most comprehensive approach available and utilizes information available for some species to prioritize genes in a wide range of organisms. In the third part of the thesis, I applied this system to the set of candidate genes targeted by RNAi in the worm and demonstrate GPSy s ability to efficiently discriminate positive candidates from those showing no phenotype. I also tested the use of this tool in the mammalian context by prioritizing a list of genes identified by expression profiling of infertile patients. The justification of several high- and low-ranking genes, both in the human and the worm datasets, suggests that the system is scientifically sound and stresses the importance of a cross-species approach. Integrative genomics approaches, such as those presented in this manuscript, will play a decisive role in the identification of genes important for spermatogenesis and reproductive disorders.Les progrès récents dans les technologies de génomique et à haut débit ont considérablement contribué à une meilleure compréhension de la spermatogenèse par l'identification de nombreux gènes exprimés au cours de la différenciation des cellules de la lignée germinale. Néanmoins, ces approches ne permettent pas d'identifier directement des gènes essentiels pour ce processus et nécessitent l'intégration de plusieurs stratégies pour la sélection des gènes candidats avant d'envisager des études fonctionnelles. La première partie de ma thèse a consisté en un crible de données d'expression basé sur la conservation du transcriptome testiculaire entre trois espèces de mammifères. Par la suite, les homologues des gènes candidats sélectionnés ont été inactivés dans l'organisme modèle C. elegans afin d'examiner d'éventuels phénotypes liés à une infertilité. La deuxième partie détaille la construction d'un outil pour la priorisation de gènes candidats, GPSy (Gene Prioritization System), basée sur l'exploitation de multiples données, parmi lesquels des données phénotypiques. Le système implémenté a pour objectif d'ordonner les gènes en fonction de leur importance relative au regard du processus de la spermatogenèse par l'intégration de plusieurs types d'informations génomiques. GPSy est actuellement l'approche la plus exhaustive disponible basée sur la transversalité des informations et données disponibles au travers d'un large éventail d'organismes (inter-espèces). Dans la troisième partie de la thèse, j'ai appliqué ce système à l'ensemble des gènes candidats sélectionnés pour le criblage par RNAi chez C. elegans. J'ai ainsi pu démontrer la capacité de GPSy à discriminer efficacement les candidats positifs de ceux ne montrant aucun phénotype. L'utilité de cet outil a par la suite été testée chez l'homme afin de prioriser une liste de gènes montrant un différentiel d'expression dans des échantillons de biopsies testiculaires de patients infertiles. L'ensemble des résultats et des discussions associées, à la fois chez le ver et l'homme, soulignent la robustesse du système développé et l'importance d'une approche inter-espèces. A l'avenir, les approches de génomique intégrative, telles que celles présentées dans ce manuscrit, seront un atout décisif pour l'identification de gènes importants pour la spermatogenèse et la compréhension fine de ce processus complexe.RENNES1-BU Sciences Philo (352382102) / SudocSudocFranceF

Apoptosis induction by activator protein 2α involves transcriptional repression of Bcl-2

Activator protein 2α (AP-2α) induces cytotoxicity by inducing cell cycle arrest and apoptosis. In this study we investigated the mechanism of apoptosis induction by AP-2α. We found that AP-2α induced apoptosis efficiently in cells treated with benzyloxycar-bonyl-IETD-fluoromethyl ketone or FADD-silenced cells but failed to do so in benzyloxycarbonyl-LEHD-fluoromethyl ketone-treated or apoptosis protease activation factor-1 (Apaf1)-silenced cells, suggesting the central role of mitochondria in AP-2α-induced apoptosis. In good correlation, cells overexpressing AP-2α showed a reduction in mitochondrial membrane potential (Δψ<SUB>m</SUB>), cytochrome c and Smac/DIABLO release into cytosol, and Bax translocation into mitochondria. We found that the pro-apoptotic protein Bax is important for AP-2α-induced apoptosis as adenovirus AP2 failed to induce apoptosis in HCT116 Bax<SUP>−/−</SUP> cells. However, we found the IAP (inhibitor of apoptosis) inhibitor Smac/DIABLO may have a limited role in AP-2α-induced apoptosis as we found the IAP member Survivin down-regulated by AP-2α. Although the total Bax level remains unaltered, we found a time-dependent increase in the activated form of Bax in adenovirus AP2-infected cells. In addition, we show that AP-2α transcriptionally represses Bcl-2 by binding to its promoter both in vitro and in vivo and that this is essential for AP-2α-induced apoptosis as ectopic expression of Bcl-2 efficiently inhibited apoptosis induced by AP-2α. Furthermore, we show that chemotherapy-induced endogenous AP-2α down-regulates Bcl-2 and induces apoptosis in an AP-2α-dependent manner. Moreover, we demonstrate that inhibition of okadaic acid or staurosporine-sensitive pathways in AP-2α overexpressing breast cancer cells resulted in AP-2α-dependent apoptosis induction. These results suggest that AP-2α induces apoptosis by down-regulating Bcl-2 and utilizing a bax/cytochrome c/Apaf1/caspase 9-dependent mitochondrial pathway

Apoptosis induction by activator Protein 2\alpha involves transcriptional repression of Bcl-2

Activator protein 2 alpha ( AP-2 alpha) induces cytotoxicity by inducing cell cycle arrest and apoptosis. In this study we investigated the mechanism of apoptosis induction by AP-2 alpha. We found that AP-2 alpha induced apoptosis efficiently in cells treated with benzyloxycarbonyl-IETD-fluoromethyl ketone or FADD silenced cells but failed to do so in benzyloxycarbonyl- EHD-fluoromethyl ketone-treated or apoptosis protease activation factor-1 ( Apaf1)-silenced cells, suggesting the central role of mitochondria in AP-2 alpha-induced apoptosis. In good correlation, cells overexpressing AP-2 alpha showed a reduction in mitochondrial membrane potential ( Delta Psi(m)), cytochrome c and Smac/DIABLOm release into cytosol, and Bax translocation into mitochondria. We found that the pro-apoptotic protein Bax is important for AP-2 alpha-induced apoptosis as adenovirus AP2 failed to induce apoptosis in HCT116 Bax(-/-) cells. However, we found the IAP ( inhibitor of apoptosis) inhibitor Smac/DIABLO may have a limited role in AP-2 alpha-induced apoptosis as we found the IAP member Survivin down-regulated by AP-2 alpha. Although the total Bax level remains unaltered, we found a time-dependent increase in the activated form of Bax in adenovirus AP2-infected cells. In addition, we show that AP-2 alpha transcriptionally represses Bcl-2 by binding to its promoter both in vitro and in vivo and that this is essential for AP-2 alpha- nduced apoptosis as ectopic expression of Bcl-2 efficiently inhibited apoptosis induced by AP-2 alpha. Furthermore, we show that chemotherapy-induced endogenous AP-2 alpha down-regulates Bcl-2 and induces apoptosis in an AP-2 alpha-dependent manner. Moreover, we demonstrate that inhibition of okadaic acid or staurosporine-sensitive pathways in AP-2 alpha overexpressing breast cancer cells resulted in AP-2 alpha-dependent apoptosis induction. These results suggest that AP-2 alpha induces apoptosis by down-regulating Bcl-2 and utilizing a bax/cytochrome c/Apaf1/caspase 9-dependent mitochondrial pathway

Cytotoxicity of half sandwich ruthenium(II) complexes with strong hydrogen bond acceptor ligands and their mechanism of action

Neutral and cationic organometallic ruthenium(II) piano stool complexes of the type [(η6-cymene)RuCl(X)(Y)] (complexes R1-R8) has been synthesized and characterized. In cationic complexes, X, Y is either a η2 phosphorus ligand such as 1,1-bis(diphenylphosphino)methane (DPPM) and 1,2-bis(diphenylphosphino)ethane (DPPE) or partially oxidized ligands such as 1,2-bis(diphenylphosphino)methane monooxide (DPPMO) and 1,2-bis(diphenylphosphino)ethane monooxide (DPPEO) which are strong hydrogen bond acceptors. In neutral complexes, X is chloride and Y is a monodentate phosphorous donor. Complexes with DPPM and DPPMO ligands ([(η6-cymene)Ru(η2-DPPM)Cl]PF6 (R2), [(η6-cymene)Ru(η2-DPPMO)Cl]PF6 (R3), [(η6-cymene)Ru(η1-DPPM)Cl2] (R5) and [(η6-cymene)Ru(η1-DPPMO)Cl2] (R6) show good cytotoxicity. Growth inhibition study of several human cancer cell lines by these complexes has been carried out. Mechanistic studies for R5 and R6 show that inhibition of cancer cell growth involves both cell cycle arrest and apoptosis induction. Using an apoptosis PCR array, we identified the sets of anti-apoptotic genes that were down regulated and pro-apoptotic genes that were up regulated. These complexes were also found to be potent metastasis inhibitors as they prevented cell invasion through matrigel. The complexes were shown to bind DNA in a non intercalative fashion and cause unwinding of plasmid DNA in cell-free medium by competitive ethidium bromide binding, viscosity measurements, thermal denaturation and gel mobility shift assays

GPSy: a cross-species gene prioritization system for conserved biological processes--application in male gamete development.

International audienceWe present gene prioritization system (GPSy), a cross-species gene prioritization system that facilitates the arduous but critical task of prioritizing genes for follow-up functional analyses. GPSy's modular design with regard to species, data sets and scoring strategies enables users to formulate queries in a highly flexible manner. Currently, the system encompasses 20 topics related to conserved biological processes including male gamete development discussed in this article. The web server-based tool is freely available at http://gpsy.genouest.org