http://sgc.anlis.gob.ar/handle/123456789/1668 2026-04-11T05:53:10Z http://sgc.anlis.gob.ar/handle/123456789/2416 Title: Bioseguridad en bioterio. Buenas prácticas de manejo, cuidado y uso de animales de laboratorio Authors: Vázquez, Luciana; Mele, Daniela; Nuñez, Héctor; Figallo, Luciano; Edelstein, Alexis; Vilardo, Ariel Esteban; Nusblat, Leonora Abstract: Este documento busca contribuir con el conocimiento de aquellos aspectos sustantivos relacionados con el trabajo en el Bioterio como la Bioética y la Bioseguridad, las Buenas Prácticas y recomendaciones en el cuidado, manejo y uso del animal de laboratorio. El objetivo propuesto es contar con un instrumento marco tanto para el personal que trabaja en bioterios de producción y experimentación animal, como para el investigador que los utiliza; del mismo modo, orienta y favorece la adopción de buenas prácticas de laboratorio, promueve la difusión de conocimientos y la asunción de responsabilidades. 2011-01-01T00:00:00Z http://sgc.anlis.gob.ar/handle/123456789/2248 Title: Optimizing the transient transfection process of HEK-293 suspension cells for protein production by nucleotide ratio monitoring Authors: Bassani Molinas, María; Beer, Christiane; Hesse, Friedemann; Wirth, Manfed; Wagner, Roland Abstract: Large scale, transient gene expression (TGE) is highly dependent of the physiological status of a cell line. Therefore, intracellular nucleotide pools and ratios were used for identifying and monitoring the optimal status of a suspension cell line used for TGE. The transfection efficiency upon polyethyleneimine (PEI)-mediated transient gene delivery into HEK-293 cells cultured in suspension was investigated to understand the effect of different culture and transfection conditions as well as the significance of the culture age and the quality of the cell line used. Based on two different bicistronic model plasmids expressing the human erythropoietin gene (rHuEPO) in the first position and green fluorescent protein as reporter gene in the second position and vice versa, a completely serum-free transient transfection process was established. The process makes use of a 1:1 mixture of a special calcium-free DMEM and the FreeStyle™ 293 Expression Medium. Maximum transfectability was achieved by adjusting the ratio for complex formation to one mass part of DNA and three parts of PEI corresponding to an N/P (nitrogen residues/DNA phosphates) ratio of 23 representing a minimum amount of DNA for the polycation-mediated gene delivery. Applying this method, maximum transfectabilities between 70 and 96 % and a rHuEPO concentration of 1.6 μg mL(-1) 72 h post transfection were reached, when rHuEPO gene was expressed from the first position of the bicistronic mRNA. This corresponded to 10 % of the total protein concentration in the cell-free supernatant of the cultures in protein-free medium. Up to 30 % higher transfectabilities were found for cells of early passages compared to those from late passages under protein-free culture conditions. In contrast, when the same cells were propagated in serum-containing medium, higher transfectabilities were found for late-passage cells, while up to 40 % lower transfectabilities were observed for early-passage cells. Nucleotide pools were measured during all cell cultivations and the nucleoside triphosphate/uridine ratios were calculated. These 'nucleotide ratios' changed in an age-dependent manner and could be used to distinguish early- from late-passage cells. The observed effects were also dependent on the presence of serum in the culture. Nucleotide ratios were shown being applied to investigate the optimal passage number of cultured cell lines for achieving a maximum productivity in cultures used for transient gene expression. Furthermore, these nucleotide ratios proved to be different for transfected and untransfected cells, providing a high potential tool to monitor the status of transfection under various culture conditions. Description: Fil: Bassani Molinas, María de los Milagros. ANLIS Dr.C.G.Malbrán. Unidad Operativa Centro de Contención Biológica; Argentina.; Fil: Beer, Christiane. Aarhus University. Institute for Molecular Biology; Dinamarca.; Fil: Hesse, Friedemann. Biberach University of Applied Sciences. Cell Culture Technology; Alemania.; Fil: Wirth, Manfed. Helmholtz Centre for Infection Research. Epigenetic Regulation; Alemania.; Fil: Wagner, Roland. Rentschler Biotechnologie GmbH. Bioprocess Development; Alemania. 2014-05-01T00:00:00Z http://sgc.anlis.gob.ar/handle/123456789/1804 Title: Aesculus hippocastanum L. seed extract shows virucidal and antiviral activities against respiratory syncytial virus (RSV) and reduces lung inflammation in vivo Authors: Salinas, Franco Maximiliano; Vázquez, Luciana; Gentilini, María Virginia; O Donohoe, Ailin; Regueira, Eleonora; Nabaes Jodar, Mercedes Soledad; Viegas, Mariana; Michelini, Flavia Mariana; Hermida, Gladys; Alché, Laura Edith; Bueno, Carlos Alberto Abstract: Respiratory syncytial virus (RSV) is a leading cause of lower respiratory tract disease and bronchiolitis in children worldwide. No vaccine or specific, effective treatment is currently available. β-escin is one of the main bioactive constituents of Aesculus hippocastanum L. (Hippocastanaceae) seed extract (AH), and both β-escin and AH have demonstrated a beneficial role in clinical therapy because of their anti-edematous, anti-inflammatory and antioxidative effects. Besides, we have reported that β-escin and AH show virucidal, antiviral and immunomodulatory activities against the enveloped viruses HSV-1, VSV and Dengue virus in vitro. In this study, we demonstrate that β-escin and AH have virucidal and antiviral activities against RSV, as well as NF-κB, AP-1 and cytokine modulating activities in RSV infected epithelial and macrophage cell lines in vitro. Besides, in a murine model of pulmonary RSV infection, AH treatment improves the course of acute disease, evidenced by decreased weight loss, reduced RSV lung titers, and attenuated airway inflammation. In contrast, even though β-escin showed, similarly to AH, antiviral and immunomodulatory properties in vitro, it neither reduces viral titers nor attenuates lung injury in vivo. Thus, our data demonstrate that AH restrains RSV disease through antiviral and immunomodulatory effect. Description: Fil: Salinas, Franco Maximiliano. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de virología; Argentina.; Fil: Vázquez, Luciana. ANLIS Dr.C.G.Malbrán. Unidad Operativa Centro de Contención Biológica (UOCCB); Argentina.; Fil: Gentilini, María Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET). Instituto de Medicina Traslacional, Trasplante y Bioingeniería (IMETTYB); Argentina.; Fil: O Donohoe, Ailin. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Laboratorio de Biología de Anfibios-Histologia Animal; Argentina.; Fil: Regueira, Eleonora. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Laboratorio de Biología de Anfibios-Histologia Animal; Argentina.; Fil: Nabaes Jodar, Mercedes Soledad. Hospital de Niños "Ricardo Gutierrez". Laboratorio de virología; Argentina.; Fil: Viegas, Mariana. Hospital de Niños "Ricardo Gutierrez". Laboratorio de virología; Argentina.; Fil: Michelini, Flavia Mariana. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de virología; Argentina.; Fil: Hermida, Gladys. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Laboratorio de Biología de Anfibios-Histologia Animal; Argentina.; Fil: Alché, Laura Edith. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de virología; Argentina.; Fil: Bueno, Carlos Alberto. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Química Biológica. Laboratorio de virología; Argentina. 2019-01-01T00:00:00Z