A condition-specific codon optimization approach for improved heterologous gene expression in Saccharomyces cerevisiae

All authors are with the Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St. Stop C0400, Austin, TX 78712, USA -- Hal S. Alper is with the Institute for Cellular and Molecular Biology, The University of Texas at Austin, 2500 Speedway Avenue, Austin, TX 78712, USA -- Amanda M. Lanza Current Address: Bristol-Myers Squibb, Biologics Development, 35 South Street, Hopkinton, MA 01748, USABackground: Heterologous gene expression is an important tool for synthetic biology that enables metabolic engineering and the production of non-natural biologics in a variety of host organisms. The translational efficiency of heterologous genes can often be improved by optimizing synonymous codon usage to better match the host organism. However, traditional approaches for optimization neglect to take into account many factors known to influence synonymous codon distributions. Results: Here we define an alternative approach for codon optimization that utilizes systems level information and codon context for the condition under which heterologous genes are being expressed. Furthermore, we utilize a probabilistic algorithm to generate multiple variants of a given gene. We demonstrate improved translational efficiency using this condition-specific codon optimization approach with two heterologous genes, the fluorescent protein-encoding eGFP and the catechol 1,2-dioxygenase gene CatA, expressed in S. cerevisiae. For the latter case, optimization for stationary phase production resulted in nearly 2.9-fold improvements over commercial gene optimization algorithms. Conclusions: Codon optimization is now often a standard tool for protein expression, and while a variety of tools and approaches have been developed, they do not guarantee improved performance for all hosts of applications. Here, we suggest an alternative method for condition-specific codon optimization and demonstrate its utility in Saccharomyces cerevisiae as a proof of concept. However, this technique should be applicable to any organism for which gene expression data can be generated and is thus of potential interest for a variety of applications in metabolic and cellular engineering.Chemical EngineeringInstitute for Cellular and Molecular [email protected]

Rap as a feminist work strategy

El feminismo es un tema que debería trabajarse de forma activa en la escuela, tanto primaria como secundaria. Es un tema fundamental para la formación de ciudadanas y ciudadanos responsables y comprometidos con la justicia social. Por ello presentamos una propuesta pedagógica de trabajo del feminismo en la secundaria obligatoria a través de la asignatura de música. Buscamos motivar a las y los estudiantes a reflexionar sobre las diferentes temáticas del feminismo a través del rap creado por mujeres. A través de cinco sesiones de trabajo con las y los estudiantes de 4º de la ESO analizaremos algunas letras y a partir de las ideas que surjan de ellas, los alumnos crearán sus propias letras, que expondrán ante sus compañeros, formando parte de un concurso al estilo “Slam poetry”Feminism is a topic that shoud be adressed in high schools in an active way, in primary as in secundary education. This is a key topic for citizenship education to develop students engage with social justice. That´s why we submit this pedagogical paper of working with feminism at high school through music subjetc. We want to motivate the students to reflect about the different themes that feminism studies through rap sung by woman. During the 5 sessions working with 4º ESO´s students, we will analyse a whole song lyrics and fragments of other songs. Thereafter the ideas that arise from them, they will create their own rap, that they will show their classmates, being part of a contest in a way of “slam poetry

Women Staff In Business Schools: An Exploratory Study

This paper presents the findings of a study which aimed to identify equity issues in the working lives of women staff in a New Zealand university business school. A participatory action research methodology was adopted which includes several stages. This paper reports on the first two: focus groups and a survey. The findings suggest that over-work and lack of community are key issues. Some women feel disadvantaged in relation to male colleagues, while others reject this notion. The paper links challenges in carrying out the research to the findings themselves. Distinctive features of the project are the emphasis on overall quality of work life and the inclusion of all women staff (general/professional and academic)

Panorama actual del consumo de sodio en Colombia y los retos que presentan los ingenieros de alimentos para mitigar el desarrollo de enfermedades cardiovasculares generadas por una elevada ingesta de este componente.

En nuestra población colombiana se presenta un gran consumo de alimentos procesados aumentando el riesgo de padecer enfermedades crónicas no trasmisibles. Siendo estos alimentos de fácil acceso para el consumidor. el Modelo de perfil de nutrientes se dedica a valorar los productos procesados que son los que normalmente contienen cantidades elevadas de sodio, los efectos de la ingesta excesiva del sodio en nuestros alimentos y los principales riesgos en la salud de los consumidores por su alto contenido, causando algunas enfermedades en la población. En los últimos años atreves de anuncios por medios de comunicación se ha difundido información a la comunidad acerca de la seguridad en la elección de los alimentos industrializados, el cual tiene como objetivo fomentar una visión crítica sobre la prevención de la salud y el mejoramiento de los proceso de algunos alimentos que tiene en gran cantidad de sodio entre los cual encontramos productos lácteos, cereales y cárnicos, entre otros, dando como resultado afectaciones en la salud y la aparición de diversas enfermedades crónicas en todo el mundo, especialmente relacionadas con nutrición tales como la diabetes, la obesidad, el cáncer y enfermedades cardiovasculares, se ha resaltado la importancia de campañas de comunicación masiva en los medios de comunicación acerca de la prevención y comportamiento de las enfermedades cardiovasculares. La organización Mundial de la Salud, ha realizado investigaciones acerca de la ingesta alimentaria de sodio, en los cuales se ve afectado los niveles de tensión arterial y realiza recomendaciones en el consumo con la finalidad de reducir el riesgo de cardiopatías coronaria y accidentes cerebrovasculares isquémico y hemorrágico, dando a conocer la importancia de unas ingestas diarias no superiores a 2 g de sodio pueden ser benéficas para reducir los niveles de tensión arterial, lo cual equivale a restringir el consumo diario de sodio a menos de 5 g diarios la organización mundial ha definido estrategias siendo la más efectivas la reducción que define los límites de contenido de sodio en alimentos Estas alternativas buscan en los productos reducir el contenido de sodio en los alimentos que tengan un alto contenido, sin embargo, en la población colombiana, a través de la normatividad exige límites y plazos en la reducción de contenido de sodio, teniendo en cuenta aspectos importantes tales como tener criterios claros en la selección de categoría y factibilidad tecnológica de reducción del sodio relacionado con los aspectos de inocuidad y que los mismos sean sostenibles.In our Colombian population there is a high consumption of processed foods increasing the risk of suffering from chronic non-transmissible diseases. These foods are easily accessible to the consumer. the Nutrient Profile Model is dedicated to assessing processed products that are those that normally contain high amounts of sodium, the effects of excessive sodium intake on our food and the main health risks to consumers due to its high content, causing some diseases in the population. In recent years, information has been disseminated to the community about the safety of industrialized food choices, which aims to promote a critical vie

The role of entrepreneurial leadership of principals in high performing schools

This dissertation builds on and contributes to work in the field of entrepreneurial leadership in the educational environment. Entrepreneurial leadership is a distinctive type of leadership behaviour dealing with challenges and crises in an organisation. It focuses on entrepreneurial customs such as being a visionary thinker, innovativeness, motivation, risk-taking and being pro-active. Although several studies have examined entrepreneurial leadership in schools, limited research exists on the impact of the school principal’s entrepreneurial leadership and management practice and the performance of the school. The purpose of this dissertation is to examine the role that entrepreneurial leadership practices play in the leadership and management practices of high performing schools as best practice study. It aims to highlight those practices in the leadership and management of a school principal which may contribute positively to the quality of education in South Africa. The data for this qualitative study were collected through open-ended, semi-structured interviews with the principals of the top 10 high performing schools in Gauteng based on their Grade 12 academic results in 2015 and 2016. A deductive case study design was considered as most suited for this research as it aimed to gaining better insight and understanding of the leadership practices of the school principals of high performing schools in real-life context. For this study, content analysis was employed. The sources of raw data were the transcripts of the individual semi-structured interviews conducted with the principals in high performing schools. The research results indicated that all principals participating in the study showed attributes of applying entrepreneurial leadership practices and incidentally used entrepreneurial leadership skills fruitfully in leading and managing the schools. The dissertation draws strongly on the work of Pihie, Bagheri and Asimiran. They are some of the first researchers who focused on entrepreneurial leadership practice and school innovativeness which empower educational leaders to meet the diverse needs of learners as well as the ever-changing demands of the school environment.Dissertation (MEd)--University of Pretoria, 2018.Education Management and Policy StudiesMEdUnrestricte

Long-Term Fructose Intake Increases Adipogenic Potential: Evidence of Direct Effects of Fructose on Adipocyte Precursor Cells

We have previously addressed that fructose rich diet (FRD) intake for three weeks increases the adipogenic potential of stromal vascular fraction cells from the retroperitoneal adipose tissue (RPAT). We have now evaluated the effect of prolonged FRD intake (eight weeks) on metabolic parameters, number of adipocyte precursor cells (APCs) and in vitro adipogenic potential from control (CTR) and FRD adult male rats. Additionally, we have examined the direct fructose effects on the adipogenic capacity of normal APCs. FRD fed rats had increased plasma levels of insulin, triglyceride and leptin, and RPAT mass and adipocyte size. FACS studies showed higher APCs number and adipogenic potential in FRD RPAT pads; data is supported by high mRNA levels of competency markers: PPARγ2 and Zfp423. Complementary in vitro experiments indicate that fructose-exposed normal APCs displayed an overall increased adipogenic capacity. We conclude that the RPAT mass expansion observed in eight week-FRD fed rats depends on combined accelerated adipogenesis and adipocyte hypertrophy, partially due to a direct effect of fructose on APCs.Facultad de Ciencias Médica

M1 macrophage subtypes activation and adipocyte dysfunction worsen during prolonged consumption of a fructose-rich diet

Fructose-rich diet (FRD) has been associated with obesity development, which is characterized by adipocytes hypertrophy and chronic low-grade inflammation. Interaction of adipocytes and immune cells plays a key role in adipose tissue (AT) alterations in obesity. We assessed the metabolic and immune impairments in AT in a murine obesity model induced by FRD at different periods. Adult Swiss mice were divided into groups of 6 and 10 weeks of fructose (FRD 6wk, FRD 10wk) or water intake (CTR 6wk, CTR 10wk). FRD induced increased in body weight, epidydimal AT mass, and plasmatic and liver Tg, and impaired insulin sensitivity. Also, hypertrophic adipocytes from FRD 6wk-10wk mice showed higher IL-6 when stimulated with LPS and leptin secretion. Several of these alterations worsened in FRD 10wk. Regarding AT inflammation, FRD mice have increased TNFα, IL-6 and IL1β, and decrease in IL-10 and CD206 mRNA levels. Using CD11b, LY6C, CD11c and CD206 as macrophages markers, we identified for first time in AT M1 (M1a: Ly6C+/−CD11c+CD206− and M1b: Ly6C+/−CD11c+CD206+) and M2 subtypes (Ly6C+/−CD11c−CD206+). M1a phenotype increased from 6 weeks onward, while Ly6C+/− M1b phenotype increased only after 10 weeks. Finally, co-culture of RAW264.7 (monocytes cell line) and CTR or FRD adipocytes showed that FRD 10wk adipocytes increased IL-6 expression in non- or LPS-stimulated monocytes. Our results showed that AT dysfunction got worse as the period of fructose consumption was longer. Inflammatory macrophage subtypes increased depending on the period of FRD intake, and hypertrophic adipocytes were able to create an environment that favored M1 phenotype in vitro.Fil: Gambaro, Sabrina Eliana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Zubiría, María Guillermina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentina. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas; ArgentinaFil: Portales, Andrea Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Rey, Maria Amanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; ArgentinaFil: Rumbo, Martín. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Estudios Inmunológicos y Fisiopatológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Estudios Inmunológicos y Fisiopatológicos; ArgentinaFil: Giovambattista, Andres. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Departamento de Ciencias Biológicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto Multidisciplinario de Biología Celular. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Instituto Multidisciplinario de Biología Celular. Universidad Nacional de La Plata. Instituto Multidisciplinario de Biología Celular; Argentin

Deleterious Metabolic Effects of High Fructose Intake: The Preventive Effect of Lactobacillus kefiri Administration

Modern lifestyle and diets have been associated with metabolic disorders and an imbalance in the normal gut microbiota. Probiotics are widely known for their health beneficial properties targeting the gut microbial ecosystem. The aim of our study was to evaluate the preventive effect of Lactobacillus kefiri (L. kefiri) administration in a fructose-rich diet (FRD) mice model. Mice were provided with tap water or fructose-added (20% w/v) drinking water supplemented or not with L. kefiri. Results showed that probiotic administration prevented weight gain and epidydimal adipose tissue (EAT) expansion, with partial reversion of the adipocyte hypertrophy developed by FRD. Moreover, the probiotic prevented the increase of plasma triglycerides and leptin, together with the liver triglyceride content. Leptin adipocyte secretion was also improved by L. kefiri, being able to respond to an insulin stimulus. Glucose intolerance was partially prevented by L. kefiri treatment (GTT) and local inflammation (TNFα; IL1β; IL6 and INFγ) was completely inhibited in EAT. L. kefiri supplementation generated an impact on gut microbiota composition, changing Bacteroidetes and Firmicutes profiles. Overall, our results indicate that the administration of probiotics prevents the deleterious effects of FRD intake and should therefore be promoted to improve metabolic disorders.Facultad de Ciencias Exacta

Hypogammaglobulinaemia secondary to cow-milk allergy in children under 2 years of age

Symptomatic hypogammaglobulinaemia in children younger than 2 years of age was studied to rule out a primary immunodeficiency. Thirty-four patients were referred to the Immunology Service to study the hypogammaglobulinaemia- associated clinical picture. Food allergy was documented in 10 patients by personal and familial history, presence of specific immunoglobulin E (IgE) and elevated total serum IgE levels. Coeliac disease and human immunodeficiency virus infection were also ruled out. Protein loss through stools was assessed by clearance of α1-antitrypsin (AAT). Serum immunoglobulin levels were determined by nephelometry and functional antibodies were studied by enzyme-linked immunosorbent assay. The cellular immune response was assessed by in vitro lymphocyte proliferation in response to mitogens and cell subsets were analysed by flow cytometry. In five patients of the 10 patients we suspected a protein loss through the mucosa. Four of these five patients showed an increased AAT and the other showed an extensive cutaneous lesion. Immunological studies revealed normal antibody function, in vitro lymphoproliferative responses and cell numbers in four of the 5 patients. One patient showed abnormally low numbers of CD4+ T cells as well as a defective proliferative response to mitogens. After diagnosis of cow milk allergy, milk was replaced with infant milk formula containing hydrolysed proteins. Recovery of immunoglobulin values and clinical resolution were achieved. Hypogammaglobulinaemia during early childhood in some children may be secondary to cow milk allergy, and immunoglobulins and cells may leak through the inflamed mucosa. Resolution of symptoms as well as normalization of immunoglobulin values may be easily achieved by avoidance of the offending allergen.Laboratorio de Investigaciones del Sistema Inmun

Intervención en diferentes instancias de la secuencia didáctica a través de experiencias de laboratorio como herramientas para la comprensión

Se intervino en 3 instancias de la secuencia didáctica con experiencias de laboratorio en distintos cursos con el propósito de mejorar la comprensión del “concepto de densidad”, el cual resulta de difícil comprensión por parte de los estudiantes de 2º año de química. Se analizaron los distintos grados de comprensión que estimularon los trabajos prácticos introducidos en distintos momentos del aprendizaje. La ponderación de los niveles de desempeño alcanzados por los distintos cursos fue realizada a través de una actividad de integración. La intervención del trabajo práctico como herramienta para la comprensión al final de la secuencia didáctica mostró mejores resultados con respecto al objetivo planteado.Facultad de Humanidades y Ciencias de la Educació