Metabolic and Kinetic analyses of influenza production in perfusion HEK293 cell culture

<p>Abstract</p> <p>Background</p> <p>Cell culture-based production of influenza vaccine remains an attractive alternative to egg-based production. Short response time and high production yields are the key success factors for the broader adoption of cell culture technology for industrial manufacturing of pandemic and seasonal influenza vaccines. Recently, HEK293SF cells have been successfully used to produce influenza viruses, achieving hemagglutinin (HA) and infectious viral particle (IVP) titers in the highest ranges reported to date. In the same study, it was suggested that beyond 4 × 10⁶cells/mL, viral production was limited by a lack of nutrients or an accumulation of toxic products.</p> <p>Results</p> <p>To further improve viral titers at high cell densities, perfusion culture mode was evaluated. Productivities of both perfusion and batch culture modes were compared at an infection cell density of 6 × 10⁶cells/mL. The metabolism, including glycolysis, glutaminolysis and amino acids utilization as well as physiological indicators such as viability and apoptosis were extensively documented for the two modes of culture before and after viral infection to identify potential metabolic limitations. A 3 L bioreactor with a perfusion rate of 0.5 vol/day allowed us to reach maximal titers of 3.3 × 10¹¹IVP/mL and 4.0 logHA units/mL, corresponding to a total production of 1.0 × 10¹⁵IVP and 7.8 logHA units after 3 days post-infection. Overall, perfusion mode titers were higher by almost one order of magnitude over the batch culture mode of production. This improvement was associated with an activation of the cell metabolism as seen by a 1.5-fold and 4-fold higher consumption rates of glucose and glutamine respectively. A shift in the viral production kinetics was also observed leading to an accumulation of more viable cells with a higher specific production and causing an increase in the total volumetric production of infectious influenza particles.</p> <p>Conclusions</p> <p>These results confirm that the HEK293SF cell is an excellent substrate for high yield production of influenza virus. Furthermore, there is great potential in further improving the production yields through better control of the cell culture environment and viral production kinetics. Once accomplished, this cell line can be promoted as an industrial platform for cost-effective manufacturing of the influenza seasonal vaccine as well as for periods of peak demand during pandemics.</p

Anti-CD25 antibody-mediated depletion of effector T cell populations enhances susceptibility of mice to acute but not chronic Toxoplasma gondii infection.

Natural regulatory T cells (Tregs) constitutively express the IL-2R alpha-chain (CD25) on their surface. Consequently, administration of anti-CD25 Abs is a commonly used technique to deplete Treg populations in vivo. However, activated effector T cells may also transiently express CD25, and are thus also potential targets for anti-CD25 Abs. In this study using Toxoplasma gondii as a model proinflammatory infection, we have examined the capacity of anti-CD25 Abs to target effector T cell populations during an inflammatory episode, to determine to what extent that this action may modulate the outcome of disease. Anti-CD25 Ab-treated C57BL/6 mice displayed significantly reduced CD4(+) T cell IFN-gamma production during acute T. gondii infection and exhibited reduced weight loss and liver pathology during early acute infection; aspects of infection previously associated with effector CD4(+) T cell responses. In agreement, anti-CD25 Ab administration impaired parasite control and caused mice to succumb to infection during late acute/early chronic stages of infection with elevated tissue parasite burdens. In contrast, anti-CD25 Ab treatment of mice with established chronic infections did not markedly affect brain parasite burdens, suggesting that protective T cell populations do not express CD25 during chronic stages of T. gondii infection. In summary, we have demonstrated that anti-CD25 Abs may directly abrogate effector T cell responses during an inflammatory episode, highlighting important limitations of the use of anti-CD25 Ab administration to examine Treg function during inflammatory settings

Predictors of Growth and Distress Following Childhood Parentification: A Retrospective Exploratory Study

Abstract While most of the existing literature has focused on the risks associated with parentification, we examined the potential benefits (i.e., posttraumatic growth) associated with parentification. Predictors of posttraumatic growth explored in our study included: attachment, differentiation of self, parentification, and resiliency. Partial support was found for the predictor variables leading to posttraumatic growth, with resilience emerging as the strongest predictor; resilience explained 14% of the variance in posttraumatic growth. These findings suggest that future research might explore additional resiliency factors that explain positive psychological outcomes related to childhood parentification

Predictors of Growth and Distress Following Childhood Parentification: A Retrospective Exploratory Study

Abstract While most of the existing literature has focused on the risks associated with parentification, we examined the potential benefits (i.e., posttraumatic growth) associated with parentification. Predictors of posttraumatic growth explored in our study included: attachment, differentiation of self, parentification, and resiliency. Partial support was found for the predictor variables leading to posttraumatic growth, with resilience emerging as the strongest predictor; resilience explained 14% of the variance in posttraumatic growth. These findings suggest that future research might explore additional resiliency factors that explain positive psychological outcomes related to childhood parentification

Reflexiones de estudiantes universitarios, docentes de Agroecología (U.N.Cuyo) y agentes de extensión del INTA en prácticas de intervención socioproductivas en las zonas de Luján de Cuyo y Valle de Uco, Mendoza

En el marco de la asignatura Agroecología y Ambientes Rurales de la carrera en Ingeniería en Recursos Naturales Renovables (FCA- UNCuyo), se han realizado durante 10 años académicos consecutivos, dos actividades consistentes en visitar a diferentes tipos sociales agrarios ubicados en Luján de Cuyo y el Valle de Uco. En las nombradas actividades, los estudiantes realizan una visita extendida a productores que integran diferentes cadenas productivas, con el objetivo de caracterizar el ecosistema, sociosistema y tecnosistema en cada caso, además de diferenciar manejos convencionales y agroecológicos. Con la información recabada y reflexionada, los estudiantes realizan un trabajo integrador, con carácter de condición obligatoria para rendir el examen final de la asignatura, que rescata una problemática detectada y propone una solución socio tecnológica con base agroecológica, ajustada a las características de tipo social agrario abordado. El nombrado material queda a disposición de los actores involucrados para ser utilizado como una herramienta de comprensión de la dinámica socio-productiva y territorial. En el presente relato realizamos una breve síntesis sobre las experiencias de cada visita a campo y se presentan las reflexiones de los actores involucrados acerca de esta experiencia.In the framework of agroecology and rural environments race course in Engineering in Renewable Natural Resources (FCA UNCuyo) were performed for 10 consecutive school years, two consistent activities visit different agricultural social types are located in Lujan de Cuyo and Uco Valley. In the named activities, students made an extended visit to producers that integrate different production chains, in order to characterize the ecosystem, sociosistema and Technosystem in each case, in addition to conventional agro differentiate and handling. With the information gathered and reflected, students take an integrative work, as an obligatory condition to take the final exam of the subject, who rescues a detected problem and proposes a solution partner agroecologically technological basis and adjusted to the characteristics of a social nature agricultural addressed. The named material is available to the actors involved to be used as a tool for understanding the socio-productive and territorial dynamics. In this story is a brief summary of experiences of each field visit and reflections of stakeholders about this experience are presented.Eje: A2: Paisajes, Territorios y AgroecologíaFacultad de Ciencias Agrarias y Forestale

Reflexiones de estudiantes universitarios, docentes de Agroecología (U.N.Cuyo) y agentes de extensión del INTA en prácticas de intervención socioproductivas en las zonas de Luján de Cuyo y Valle de Uco, Mendoza

En el marco de la asignatura Agroecología y Ambientes Rurales de la carrera en Ingeniería en Recursos Naturales Renovables (FCA- UNCuyo), se han realizado durante 10 años académicos consecutivos, dos actividades consistentes en visitar a diferentes tipos sociales agrarios ubicados en Luján de Cuyo y el Valle de Uco. En las nombradas actividades, los estudiantes realizan una visita extendida a productores que integran diferentes cadenas productivas, con el objetivo de caracterizar el ecosistema, sociosistema y tecnosistema en cada caso, además de diferenciar manejos convencionales y agroecológicos. Con la información recabada y reflexionada, los estudiantes realizan un trabajo integrador, con carácter de condición obligatoria para rendir el examen final de la asignatura, que rescata una problemática detectada y propone una solución socio tecnológica con base agroecológica, ajustada a las características de tipo social agrario abordado. El nombrado material queda a disposición de los actores involucrados para ser utilizado como una herramienta de comprensión de la dinámica socio-productiva y territorial. En el presente relato realizamos una breve síntesis sobre las experiencias de cada visita a campo y se presentan las reflexiones de los actores involucrados acerca de esta experiencia.In the framework of agroecology and rural environments race course in Engineering in Renewable Natural Resources (FCA UNCuyo) were performed for 10 consecutive school years, two consistent activities visit different agricultural social types are located in Lujan de Cuyo and Uco Valley. In the named activities, students made an extended visit to producers that integrate different production chains, in order to characterize the ecosystem, sociosistema and Technosystem in each case, in addition to conventional agro differentiate and handling. With the information gathered and reflected, students take an integrative work, as an obligatory condition to take the final exam of the subject, who rescues a detected problem and proposes a solution partner agroecologically technological basis and adjusted to the characteristics of a social nature agricultural addressed. The named material is available to the actors involved to be used as a tool for understanding the socio-productive and territorial dynamics. In this story is a brief summary of experiences of each field visit and reflections of stakeholders about this experience are presented.Eje: A2: Paisajes, Territorios y AgroecologíaFacultad de Ciencias Agrarias y Forestale

Reflexiones de estudiantes universitarios, docentes de Agroecología (U.N.Cuyo) y agentes de extensión del INTA en prácticas de intervención socioproductivas en las zonas de Luján de Cuyo y Valle de Uco, Mendoza

En el marco de la asignatura Agroecología y Ambientes Rurales de la carrera en Ingeniería en Recursos Naturales Renovables (FCA- UNCuyo), se han realizado durante 10 años académicos consecutivos, dos actividades consistentes en visitar a diferentes tipos sociales agrarios ubicados en Luján de Cuyo y el Valle de Uco. En las nombradas actividades, los estudiantes realizan una visita extendida a productores que integran diferentes cadenas productivas, con el objetivo de caracterizar el ecosistema, sociosistema y tecnosistema en cada caso, además de diferenciar manejos convencionales y agroecológicos. Con la información recabada y reflexionada, los estudiantes realizan un trabajo integrador, con carácter de condición obligatoria para rendir el examen final de la asignatura, que rescata una problemática detectada y propone una solución socio tecnológica con base agroecológica, ajustada a las características de tipo social agrario abordado. El nombrado material queda a disposición de los actores involucrados para ser utilizado como una herramienta de comprensión de la dinámica socio-productiva y territorial. En el presente relato realizamos una breve síntesis sobre las experiencias de cada visita a campo y se presentan las reflexiones de los actores involucrados acerca de esta experiencia.In the framework of agroecology and rural environments race course in Engineering in Renewable Natural Resources (FCA UNCuyo) were performed for 10 consecutive school years, two consistent activities visit different agricultural social types are located in Lujan de Cuyo and Uco Valley. In the named activities, students made an extended visit to producers that integrate different production chains, in order to characterize the ecosystem, sociosistema and Technosystem in each case, in addition to conventional agro differentiate and handling. With the information gathered and reflected, students take an integrative work, as an obligatory condition to take the final exam of the subject, who rescues a detected problem and proposes a solution partner agroecologically technological basis and adjusted to the characteristics of a social nature agricultural addressed. The named material is available to the actors involved to be used as a tool for understanding the socio-productive and territorial dynamics. In this story is a brief summary of experiences of each field visit and reflections of stakeholders about this experience are presented.Eje: A2: Paisajes, Territorios y AgroecologíaFacultad de Ciencias Agrarias y Forestale

Sustentabilidad vitivinícola para diferentes tipos de agricultores vitivinícolas

Un Desarrollo Rural Sustentable requiere de sistemas agrícolas sustentables, que sean ecológicamente adecuados, económicamente viables y socio culturalmente aceptables Abbona et al, (2004). Sarandon, (2010), define la agricultura sustentable como aquella que es: suficientemente productiva, económicamente viable, ecológicamente adecuada y cultural y socialmente aceptable. En el contexto de cambio de la vitivinicultura, se hace necesario conocer la sustentabilidad de los mismos. En este caso se usó la tipología que propuesta por Martín, (2009) que plantea tres tipos sociales: productores descapitalizados pluriactivos, productores familiares capitalizados y empresarios. El objetivo de este trabajo fue: analizar, mediante el método propuesto por Sarandón (2002) y el uso de indicadores, la sustentabilidad predial de tres modelos productivos vitivinícolas: productores descapitalizados pluriactivos, productores familiares capitalizados y empresarios, de la 1º zona de Mendoza. La propuesta de metodológica estuvo basada en la metodología de Sarandón (2002) y estaba propuesto realizar tres etapas, de 2 años cada una. En una primera etapa se desarrollaron dos acciones. Lo primero fué buscar y analizar antecedentes e información regional sobre el tema y la segunda acción fue buscar y seleccionar 2 fincas del departamento de Lujan de Cuyo que fueran representativas, para cada tipo social, factibilidad de acceso y predisposición de los agricultores. En una segunda etapa se caracterizaría, a través del enfoque sistémico, el funcionamiento de los agroecosistemas en tres dimensiones: ambiental-ecológica, tecnológica, y socioeconómica. Y en una tercera etapa: se construirán indicadores para evaluar la sustentabilidad de los sistemas agrícolas, Se logró la primera etapa y se pudo: conceptualizar y profundizar el concepto de sustentabilidad integral y su aplicación en la agronomía, proponer variables con potencial de indicadores de sustentabilidad y seleccionar las fincas.A Sustainable Rural Development requires sustainable agricultural systems that are environmentally suitable, economically viable and culturally acceptable (Abbona et al, 2004). Sarandon (2010) defines sustainable agriculture as one that is: productive enough, affordable, ecologically appropriate and culturally and socially acceptable. In the context of the viticulture changes, it is necessary to study their sustainability. In this case we take the typology proposed by Martin, 2009 that summarizes it in three social types: pluriactive undercapitalized producers, family farmers and entrepreneurs capitalized. The objective of this work was to analyze, through the method proposed by Sarandón (2002) and the use of indicators, the property sustainability of three vitivinícolas production models: pluriactively de-capitalized producers, capitalized family producers and entrepreneurs, from the first zone of Mendoza. The methodological proposal was based on the methodology of Sarandon (2002) and was proposed to perform three stages, each of 2 years. In the first stage two actions were developed. The first was to seek and analyze regional background and information on the subject; And the second action was to search and select 2 farms in the department of Lujan de Cuyo that were representative for each social type, feasibility of access and predisposition of farmers. In a second stage, the systemic approach would characterize the functioning of agroecosystems in three dimensions: environmental-ecological, technological, and socioeconomic. And in a third stage: indicators will be constructed to evaluate the sustainability of agricultural systems, The first stage was achieved and it was possible to: conceptualize and deepen the concept of integral sustainability and its application in agronomy, propose variables with potential for sustainability indicators and successful selection of farms.Trabajo publicado en Filippini, María Flavia (comp.). Congreso Internacional Aguas, Ambiente y Energía de la Asociación de Universidades Grupo Montevideo: Resúmenes de trabajos. Mendoza: Universidad Nacional de Cuyo, 2017.Facultad de Ciencias Agrarias y Forestale

Sustentabilidad vitivinícola para diferentes tipos de agricultores vitivinícolas

Un Desarrollo Rural Sustentable requiere de sistemas agrícolas sustentables, que sean ecológicamente adecuados, económicamente viables y socio culturalmente aceptables Abbona et al, (2004). Sarandon, (2010), define la agricultura sustentable como aquella que es: suficientemente productiva, económicamente viable, ecológicamente adecuada y cultural y socialmente aceptable. En el contexto de cambio de la vitivinicultura, se hace necesario conocer la sustentabilidad de los mismos. En este caso se usó la tipología que propuesta por Martín, (2009) que plantea tres tipos sociales: productores descapitalizados pluriactivos, productores familiares capitalizados y empresarios. El objetivo de este trabajo fue: analizar, mediante el método propuesto por Sarandón (2002) y el uso de indicadores, la sustentabilidad predial de tres modelos productivos vitivinícolas: productores descapitalizados pluriactivos, productores familiares capitalizados y empresarios, de la 1º zona de Mendoza. La propuesta de metodológica estuvo basada en la metodología de Sarandón (2002) y estaba propuesto realizar tres etapas, de 2 años cada una. En una primera etapa se desarrollaron dos acciones. Lo primero fué buscar y analizar antecedentes e información regional sobre el tema y la segunda acción fue buscar y seleccionar 2 fincas del departamento de Lujan de Cuyo que fueran representativas, para cada tipo social, factibilidad de acceso y predisposición de los agricultores. En una segunda etapa se caracterizaría, a través del enfoque sistémico, el funcionamiento de los agroecosistemas en tres dimensiones: ambiental-ecológica, tecnológica, y socioeconómica. Y en una tercera etapa: se construirán indicadores para evaluar la sustentabilidad de los sistemas agrícolas, Se logró la primera etapa y se pudo: conceptualizar y profundizar el concepto de sustentabilidad integral y su aplicación en la agronomía, proponer variables con potencial de indicadores de sustentabilidad y seleccionar las fincas.A Sustainable Rural Development requires sustainable agricultural systems that are environmentally suitable, economically viable and culturally acceptable (Abbona et al, 2004). Sarandon (2010) defines sustainable agriculture as one that is: productive enough, affordable, ecologically appropriate and culturally and socially acceptable. In the context of the viticulture changes, it is necessary to study their sustainability. In this case we take the typology proposed by Martin, 2009 that summarizes it in three social types: pluriactive undercapitalized producers, family farmers and entrepreneurs capitalized. The objective of this work was to analyze, through the method proposed by Sarandón (2002) and the use of indicators, the property sustainability of three vitivinícolas production models: pluriactively de-capitalized producers, capitalized family producers and entrepreneurs, from the first zone of Mendoza. The methodological proposal was based on the methodology of Sarandon (2002) and was proposed to perform three stages, each of 2 years. In the first stage two actions were developed. The first was to seek and analyze regional background and information on the subject; And the second action was to search and select 2 farms in the department of Lujan de Cuyo that were representative for each social type, feasibility of access and predisposition of farmers. In a second stage, the systemic approach would characterize the functioning of agroecosystems in three dimensions: environmental-ecological, technological, and socioeconomic. And in a third stage: indicators will be constructed to evaluate the sustainability of agricultural systems, The first stage was achieved and it was possible to: conceptualize and deepen the concept of integral sustainability and its application in agronomy, propose variables with potential for sustainability indicators and successful selection of farms.Trabajo publicado en Filippini, María Flavia (comp.). Congreso Internacional Aguas, Ambiente y Energía de la Asociación de Universidades Grupo Montevideo: Resúmenes de trabajos. Mendoza: Universidad Nacional de Cuyo, 2017.Facultad de Ciencias Agrarias y Forestale