Comparative study of the in vitro and in vivo properties of two bovine herpesvirus-5 reference strains

peer reviewe

MAGE Proteins and the Regulation of E2F Pathway

Melanoma Antigens Genes (MAGE) constitutes a mutagenic family divided in two subfamilies, MAGE-I and MAGE-II, according to its tissue pattern expression. While MAGE-I in adult humans are only expressed in testis and tumors tissues, those belonging to MAGE-II subfamily are ubiquitously expressed. During the last decade, functional characterization of MAGE proteins points to a role in transcription regulation. E2F1 is a member of the E2F family and is among the transcription factors reported to be modulated by MAGE proteins. In this article we will focus on reported cases of E2F1 modulation by members of MAGE-I and MAGE-II subfamilies and the resulting biological consequences observed in normal and tumor cells.Fil: Ladelfa, Maria Fatima. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; ArgentinaFil: Monte, Martín. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentin

Estudiantes y autonomía. Aportes para superar la enseñanza del derecho

This paper starts from the premise that learning in the 21st century must adapt to the demands of modern society. Faced with the question of what society expects, or rather, demands from future graduates of the Faculty of Law of the University of Buenos Aires, it advocates for the implementation of various teaching and assessment strategies aimed at providing students with the necessary tools for better academic performance. To this end, the implementation of activities typical of professional life from the beginning of the career is encouraged, so that students can approach the professional world from the outset. In essence, it proposes a significant change in teaching methodology, advocating for the progressive implementation of activities inherent to the profession, enabling students to recognize and refine the natural skills and abilities of the profession from the earliest subjectsFil: Ladelfa, Federico. Universidad de Buenos Aires. Facultad de Derecho. Cátedra Elementos de Derecho Penal y Procesal Penal. Buenos Aires, ArgentinaEste trabajo parte de la premisa de considerar que los aprendizajes que se imparten en el siglo XXI deben adaptarse a las demandas de la sociedad moderna. Con ese punto de partida y frente a la pregunta sobre qué espera o, mejor dicho, qué demanda la sociedad a los futuros egresados de la Facultad de Derecho de la Universidad de Buenas Aires, se propone la implementación de diversas estrategias de enseñanza y de evaluación orientadas a brindar al estudiante las herramientas necesarias para lograr un mejor desempeño académico. A tal fin, se impulsa la implementación de actividades propias de la vida profesional desde el inicio de la carrera, de manera tal que el estudiante pueda aproximarse al mundo profesional desde el comienzo. En pocas palabras, se plantea un cambio significativo en la forma de enseñanza y se propone la implementación, de forma progresiva, de actividades propias de la profesión, de manera tal que el alumno pueda, desde las primeras materias, reconocer y perfeccionar las habilidades y destrezas naturales de la profesió

In vitro growth properties of bovine herpesvirus type 5 (BoHV-5) A663 strain and study of the Us3 protein function

Este trabajo de tesis contribuyo a la caracterizacion de la cepa argentina A663 de Herpesvirus Bovino tipo 5 (BoHV-5) mediante el estudio de sus propiedades replicativas in vitro y del rol de la proteina Us3. En el primer caso se realizaron cineticas de crecimiento y se evaluaron los tamanos de las placas de lisis y de infeccion de las cepas A663 y N569 de BoHV-5. La produccion total de particulas virales infectivas en funcion del tiempo fue similar entre ambas cepas, sin embargo la cepa A663 resulto menos eficiente en la liberacion de particulas virales al medio extracelular y en la dispersion celula a celula, observandose para la cepa A663 una reduccion del 90% y del 80% en el tamano de la placa de lisis y de infeccion, respectivamente. Por otro lado, se secuencio la region codificante para la proteina Us3 de la cepa A663 de BoHV-5 y se observo un alto porcentaje de homologia con las secuencias correspondientes a otras cepas de BoHV-5. Mediante las tecnicas de Northern y Western blot se demostro que el gen us3 se expresa tempranamente (desde los 30 minutos post infeccion) y de manera prolongada (hasta las 48 horas post infeccion). Los ensayos in vitro se realizaron utilizando construcciones que expresaban la proteina Us3 wt o una version mutante de esta proteina que carece de actividad de proteina quinasa, fusionadas a la proteina fluorescente ECFP (US3-ECFP y Us3K261D-ECFP). Se observo que la proteina Us3 de BoHV-5 induce el redondeamiento celular y provoca el desensamblado de las fibras de estres, siendo estos efectos independientes de su funcion de quinasa. Ademas, la proteina mutante Us3K261D disminuyo la formacion de proyecciones de celulares y modifico la localizacion subcelular de la proteina Us3 de BoHV-5. En conjunto, los resultados obtenidos permitieron conocer las propiedades replicativas in vitro de la cepa A663 de BoHV-5 y constituyeron el primer reporte sobre el estudio de la funcion de la proteina Us3 de BoHV-5.This thesis contributed to the characterization of the Argentinean bovine herpesvirus type 5 (BoHV-5) A663 strain by studying its in vitro growth properties and the function of the Us3 protein. Firstly, growth kinetics, lysis and infection plaque size assays were performed for BoHV-5 A663 and N569 strains. The total amount of infection virus produced was similar between both strains at each time assayed; however, A663 strain was less efficient than N569 strain in releasing infective viral particles to the extracellular media and in cell-to-cell spread, as shown by a reduction of 90% and 80% in its lysis and infection plaque size, respectively. On the other hand, the region encoding the Us3 protein of BoHV-5 A663 strain was sequenced and high homology was observed between it and other sequences of BoHV-5 strains. Then, early (from 30 minutes post infection) and prolong (up to 48 hours post infection) expression of us3 gene was demonstrated by Northern and Western blots assays. Finally, in vitro studies were performed with genetic constructions to express wt Us3 protein or a mutated version of this protein lacking its kinase activity, fused to the fluorescent protein ECFP (Us3-ECFP and Us3K261D-ECFP). In transient transfection assays it was observed that Us3 BoHV-5 protein induced cell rounding and stress fiber breakdown independently of its kinase activity. Besides, the point mutation in the Us3K261D protein reduced cell projections formation and modified the subcellular localization of BoHV-5 Us3 protein. Taken together, these results allowed the assessment of the in vitro growth properties of BoHV-5 A663 strain and constituted the first report of the function of the BoHV-5 Us3 protein.Fil:Ladelfa, María Fátima. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

PCLIB : a personal computer librarian utility

This Master's project addresses the issue of the need for automated configuration management tools for personal computers. After studying some major tools that are mainframe hosted, a PC hosted tool, PCLIB, has been designed and implemented that provides software configuration management functions similar to those found in the commercial tools. PC LIB is an interactive, menu-driven configuration management program that controls, identifies, and tracks changes to elements of a software project. It acts as an automated librarian by storing successive versions of an element in the PCLIB library. Once an element is entered into the PCLIB library, it cannot be accessed without going through the PCLIB program. In this way, PCLIB can control and track any movement of files into and out of the library. Any changes that are made to a library entry will create a new version of that file. PCLIB maintains an audit trail of all file movement and can generate a history report for any library member. In addition, it provides a brief on-line status of any entry and a facility for identifying the current baseline of the configured software. The PCLIB system is currently implemented to run on IBM PC/XT compatible personal computers with Microsoft's MS-Dos as the host operating system. The source code for PCLIB is in Turbo Pascal and all of the MS-Dos service calls are made via Pascal. It is designed to be as portable as possible and can be installed on various system configurations for different applications.Includes bibliographical references (page 54)California State University, Northridge. Department of Computer Science

O papel do monitor acadêmico na disciplina de leitura e produção de textos em língua portuguesa I

Este Trabalho de Conclusão de Curso tem como objetivo geral averiguar o papel do monitor acadêmico na disciplina de Leitura e Produção de Textos em Língua Portuguesa I (LPT I) ofertada aos alunos do curso de bacharelado em Letras na UFRGS. Perseguindo esse objetivo, o trabalho se divide em cinco partes. A primeira parte apresenta o lugar de onde essa pesquisa começa. A segunda traz a análise da Instrução Normativa nº 03/2013 da Prograd/Sead e uma síntese de outras pesquisas servindo de ponto de partida para a reflexão sobre o papel do monitor na universidade. A terceira parte exemplifica como é o andamento da disciplina de LPT I bem como o seu objeto de estudo. A quarta parte descreve a concepção de língua que baseia as reflexões deste trabalho a partir da Linguística da Enunciação de Émile Benveniste. A quinta parte se destina a discutir sobre o papel do monitor em LPT I, concluindo de que é possível percebê-lo em quatro diferentes posições: o monitor um pouco professor, um pouco aluno; o monitor como um modelo de aluno; o monitor em um metalugar — ensina escrita ao mesmo tempo que aprende escrita —; e o monitor como interlocutor e mediador de letramento.This bachelor’s thesis aims to make a discussion about the role of the university monitor in the discipline Leitura e Produção de textos em Língua Portuguesa I (LPT I), which is offered to Letras’s Bachelor students at UFRGS. To reach this objective, this work is divided into four parts. The first one shows where this research starts. The second one brings an analysis of the Instrução Normativa nº 03/2013 from Prograd/Sead as well as a summary of other research, serving as a starting point for the following discussion. The third part exemplifies how the LPT I classes are handled and its purpose. The fourth part describes the concept of language through Emile Benveniste’s Linguistics of Enunciation on which the following reflections are going to be based. The fifth part intends to discuss the roles of the monitor in LPT I concluding that it is possible to divide them into four different positions: the monitor as a teacher and as a student; the monitor as a student model; the monitor in a meta place — teaches writing at the same time as learns writing; and the monitor as interlocutor and mediator

MageA2 restrains cellular senescence by targeting the function of PMLIV/p53 axis at the PML-NBs

MAGE-A genes are a subfamily of the melanoma antigen genes (MAGEs), whose expression is restricted to tumor cells of different origin and normal tissues of the human germline. Although the specific function of individual MAGE-A proteins is being currently explored, compelling evidence suggest their involvement in the regulation of different pathways during tumor progression. We have previously reported that MageA2 binds histone deacetylase (HDAC)3 and represses p53-dependent apoptosis in response to chemotherapeutic drugs. The promyelocytic leukemia (PML) tumor suppressor is a regulator of p53 acetylation and function in cellular senescence. Here, we demonstrate that MageA2 interferes with p53 acetylation at PML-nuclear bodies (NBs) and with PMLIV-dependent activation of p53. Moreover, a fraction of MageA2 colocalizes with PML-NBs through direct association with PML, and decreases PMLIV sumoylation through an HDAC-dependent mechanism. This reduction in PML post-translational modification promotes defects in PML-NBs formation. Remarkably, we show that in human fibroblasts expressing RasV12 oncogene, MageA2 expression decreases cellular senescence and increases proliferation. These results correlate with a reduction in NBs number and an impaired p53 response. All these data suggest that MageA2, in addition to its anti-apoptotic effect, could have a novel role in the early progression to malignancy by interfering with PML/p53 function, thereby blocking the senescence program, a critical barrier against cell transformation. © 2012 Macmillan Publishers Limited. All rights reserved.Fil:Ladelfa, M.F. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina.Fil:Monte, M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Dengue Non-structural Protein 5 Polymerase Complexes With Promyelocytic Leukemia Protein (PML) Isoforms III and IV to Disrupt PML-Nuclear Bodies in Infected Cells

Dengue virus (DENV) threatens almost 70% of the world's population, with no therapeutic currently available. The severe, potentially lethal forms of DENV disease (dengue hemorrhagic fever/dengue shock syndrome) are associated with the production of high level of cytokines, elicited as part of the host antiviral response, although the molecular mechanisms have not been fully elucidated. We previously showed that infection by DENV serotype 2 (DENV2) disrupts promyelocytic leukemia (PML) gene product nuclear bodies (PML-NBs) after viral protein translation in infected cells. Apart from playing a key role as the nucleating agent in forming PML-NBs, PML has antiviral activity against various viruses, including DENV. The present study builds on this work, showing for the first time that all four DENV serotypes elicit PML-NB breakdown. Importantly, we show for the first time that of the nuclear localizing proteins of DENV, DENV non-structural protein (NS) 5 polymerase alone is sufficient to elicit PML-NB disassembly, in part through complexing with PML isoforms III and IV, but not other PML isoforms or other PML-NB components. The results raise the possibility that PML-NB disruption by nuclear localized NS5 contributes to DENV's suppression of the host antiviral response.Fil: Giovannoni, Federico. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ladelfa, Maria Fatima. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Monte, Martin. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; ArgentinaFil: Jans, David A.. Monash University; AustraliaFil: Hemmerich, Peter. Leibniz Institute On Aging; AlemaniaFil: Garcia, Cybele. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica; Argentin