Development of human central nervous system 3D in vitro models for preclinical research

Neurological disorders are a major concern in modern societies, with increasing prevalence mainly related with the higher life expectancy. Most of the current available therapeutic options can only control and ameliorate the patients’ symptoms, often be-coming refractory over time. Therapeutic breakthroughs and advances have been hampered by the lack of accurate central nervous system (CNS) models. The develop-ment of these models allows the study of the disease onset/progression mechanisms and the preclinical evaluation of novel therapeutics. This has traditionally relied on genetically engineered animal models that often diverge considerably from the human phenotype (developmentally, anatomically and physiologically) and 2D in vitro cell models, which fail to recapitulate the characteristics of the target tissue (cell-cell and cell-matrix interactions, cell polarity). The in vitro recapitulation of CNS phenotypic and functional features requires the implementation of advanced culture strategies that enable to mimic the in vivo struc-tural and molecular complexity. Models based on differentiation of human neural stem cells (hNSC) in 3D cultures have great potential as complementary tools in preclinical research, bridging the gap between human clinical studies and animal models. This thesis aimed at the development of novel human 3D in vitro CNS models by integrat-ing agitation-based culture systems and a wide array of characterization tools. Neural differentiation of hNSC as 3D neurospheres was explored in Chapter 2. Here, it was demonstrated that human midbrain-derived neural progenitor cells from fetal origin (hmNPC) can generate complex tissue-like structures containing functional dopaminergic neurons, as well as astrocytes and oligodendrocytes. Chapter 3 focused on the development of cellular characterization assays for cell aggregates based on light-sheet fluorescence imaging systems, which resulted in increased spatial resolu-tion both for fixed samples or live imaging. The applicability of the developed human 3D cell model for preclinical research was explored in Chapter 4, evaluating the poten-tial of a viral vector candidate for gene therapy. The efficacy and safety of helper-dependent CAV-2 (hd-CAV-2) for gene delivery in human neurons was evaluated, demonstrating increased neuronal tropism, efficient transgene expression and minimal toxicity. The potential of human 3D in vitro CNS models to mimic brain functions was further addressed in Chapter 5. Exploring the use of 13C-labeled substrates and Nucle-ar Magnetic Resonance (NMR) spectroscopy tools, neural metabolic signatures were evaluated showing lineage-specific metabolic specialization and establishment of neu-ron-astrocytic shuttles upon differentiation. Chapter 6 focused on transferring the knowledge and strategies described in the previous chapters for the implementation of a scalable and robust process for the 3D differentiation of hNSC derived from human induced pluripotent stem cells (hiPSC). Here, software-controlled perfusion stirred-tank bioreactors were used as technological system to sustain cell aggregation and dif-ferentiation. The work developed in this thesis provides practical and versatile new in vitro ap-proaches to model the human brain. Furthermore, the culture strategies described herein can be further extended to other sources of neural phenotypes, including pa-tient-derived hiPSC. The combination of this 3D culture strategy with the implemented characterization methods represents a powerful complementary tool applicable in the drug discovery, toxicology and disease modeling.

Aspectos fundamentais da experiência da vida

A tese que de seguida se esboça assenta sobre uma inquietação fundamental: o facto de cada um dar por si atirado na vida, de, quando cada um dá por si, dar por si a ser vida, etc. Acontece que, logo que se tenta focar mais precisamente de que é de que se trata quando se trata da “vida”, nota-se que esse fenómeno tem habitualmente a forma de um acontecimento anónimo: não se sabe bem a que é que corresponde, que conteúdos tem, que estruturas fundamentais a suportam, etc. Isto é: somos levados pela vida (passamos pela vida, atravessamo-la, estamos expostos a ela, etc.) sem saber exactamente a que é que estamos expostos, o que é que nos leva, sobre que pilares assenta a nossa vivência e a nossa compreensão dela, etc. A tese que se segue não tem a pretensão de deixar definitivamente respondidas estas perguntas; tudo o que faz, na verdade, é meramente proceder a um breve levantamento ou a um registo de algumas das estruturas fundamentais da vida a partir do ângulo da experiência da vida. E, como se espera deixar claro, procurar a resposta a partir do ângulo da experiência (do ângulo da experiência da vida) não é algo acidental ou fortuito. O que se procurará apurar é se não haverá tais laços de afinidade entre “vida” e “experiência” que todas as operações próprias da experiência têm lugar numa vinculação e estão subordinadas às estruturas fundamentais da vida (estruturas que ultrapassam o âmbito da “experiência”) e que, assim também, a vida tenha, de raiz, no modo como nela somos levados e conduzidos, a estrutura ou a forma da “experiência”.

Fenologia e produtividade do Infá-Cipó (Inga edulis) na Amazônia Central

O ingá-cipó (Inga edulis Martius) é uma leguminosa arbórea da sub-família Mimosoideae, nativa da América Tropical, e amplamente cultivada pela população local por fornecer fruto comestível, madeira boa para lenha, como árvore de sombra, e mais recentemente como componente agroflorestal. O estudo da fenologia do ingá-cipó ajudará a planejar a comercialização dos frutos e o manejo dos plantios. Observou-se quatro períodos de floração durante o ano, com picos em março, maio, agosto/setembro, outubro/janeiro; algumas árvores apresentaram cinco florações. Os picos de frutificação ocorreram em abril, junho, setembro/ outubro, novembro/fevereiro. Os ingá-cipós de 3-4 anos produziram de 20.000 a 100.000 flores (media de 50.000) e 200 a 800 frutos (media de 500). O vingamento dos frutos variou de 0,4 a 1,8%, com uma média geral de 1,1%. O peso dos frutos variou de 250 a 600 g (média de 470 g), contendo 22±4% de polpa comestível. A produção anual de frutos por árvore variou de 300 a 1.700 kg (média de 960 kg).

Prevention in underground construction with sequential excavation method

This paper aims to describe the Sequential Excavation Method, used for excava-tion in underground works, as well as the related risks and preventive measures. This method has characteristics that differentiate it from other tunnelling techniques: it uses a larger number of workers and equipment; it has a high concurrency of tasks with various workers and equip-ment quite exposed to hazards; and it uses many potentially aggressive chemicals. Firstly, it is given a broad overview of this issue. Afterwards, it will be presented the results of a survey to a sample of experienced technicians, aimed at gauging the relevance of a set of guidelines relat-ing to the design and work phases, applicable to the domestic market and prepared following technical visits to works abroad.

Poly-N-acetyl glucosamine expression in Staphylococcus epidermidis biofilm cells affects the immune response and promotes tissue pathology in infected hosts

Staphylococcus epidermidis is a biofilm - forming bacterium and a leading etiological agent of nosocomial infections. The ability to establish biofilms on indwelling medical devices is a key virulence factor for this bacterium. Still, the influence of poly - N - acetyl glucosamine (PNAG), the major component of the extracellular biofilm matrix, in the host immune response has been scarcely studied. Here, t h is influence was assessed in mice challenged i.p. with PNAG - p roducing (WT) and isogenic - mutant lacking PNAG (M10) bacteria grown in biofilm - inducing conditions. Faster bacterial clearance was observed in the mice infected with WT bacteria than in M10 - infected counterparts , which w as accompanied by earlier neutrophil recruitment and higher IL - 6 production. Interestingly, in the WT - infected mice, but not in those infected with M10 , elevated serum IL - 10 was detected . To further study the effe ct of PNAG in the immune response, mice were primed with WT or M10 biofilm bacteria and subsequently infected with WT biofilm - released cells. WT - primed mice presented a higher frequency of splenic IFN - γ + and IL - 17 + CD4 + T cells, and more severe liver patho logy than M10 - primed counterparts. Nevertheless, T reg cells obtained from the WT - primed mice presented a higher suppressive function than those obtained from M10 - primed mice. This effect was abrogated when IL - 10 - deficient mice were similarly primed and infected indicating that PNAG promotes the differentiati on of highly suppressive T reg cells by a mechanism dependent on IL - 10. Altogether, these results provide evidence help ing explain ing the coexistence of inflammation and bacterial persistence often observed in biofilm - originated S. epidermidis infections

Generating timed trajectories foran autonomous robot

Tese de Doutoramento Programa Doutoral em Engenharia Electrónica e Computadores

Processing and characterization of plates made from granulate of waste electrical cables

This article is intended to evaluate the density and the mechanical, acoustic and thermal properties of compression moulded plates composed of granulate from electrical cables wastes. Those cable wastes are the insulation part from the electric cables, and are composed of PVC, PE, EMP and PEX rubber. After these materiais lose their initial properties and cease to be useful as insulation material, due to safety requirements, it is possible to reuse them into new applications like industrial or playground floorings, as sound insulation material to be applied in walls or floors, or to dampen vibrations from equipments. Recovering electric cable waste has been a major concern to the European Commission due to its leveis of toxicity when incineration and land fill ing is the solution to dispose this material. Such as the European Commission's study for DG Xl[1] suggested that recycling may be the most favourable future waste management option.

“Safety and health” as a criterion in the choice of tunneling method

Risk management is of paramount importance in the success of tunnelling works and is linked to the tunnelling method and to the constraints of the works. Sequencial Excavation Method (SEM) and Tun-nel Boring Machine (TBM) method have been competing for years. This article, part of a wider study on the influence of the â Safety and Healthâ criterion in the choice of method, reviews the existing literature about the criteria usually employed to choose the tunnelling method and on the criterion â Safety and Healthâ . This crite-rion is particularly important, due to the financial impacts of work accidents and occupational diseases. This article is especially useful to the scientific and technical community, since it synthesizes the relevance of each one of the choice criteria used and it shows why â Safety and Healthâ must be a criterion in the decision mak-ing process to choose the tunnelling method.

OBSERVATORIO AMBIENTAL Y EPIDEMIOLOGICO DE POBLACIONES DE LA PROVINCIA DE CORDOBA EXPUESTAS A AGROQUÍMICOS

El cambio climático constituye una nueva e importante amenaza para la salud, y modifica la manera en que debemos considerar la protección de las poblaciones vulnerables. Por otra parte, las actividades antrópicas son las responsables de dicho cambio, por lo cual es necesario incidir en esas causas. Muchas de ellas están relacionadas con el actual modelo agrícola, que deteriora la capacidad productiva del suelo en particular y de los ecosistemas en general, erosionando la biodiversidad, destruyendo los procesos ecológicos esenciales, contaminando el suelo, el aire y el agua, generando enfermedades de origen toxicológico y siendo además el principal causante de los gases de efecto invernadero, responsable del 25% de las emisiones de bióxido de carbono y del 80% de óxido nitroso en el planeta. En Argentina las nuevas tecnologías agrícolas, han modificado profundamente el escenario socio ambiental de la república. Las poblaciones próximas a zonas sembradas están expuestas a múltiples vías de contaminación. Sin embargo, no existe a nivel regional un estudio sistemático de los parámetros ambientales que puedan alertar sobre los problemas derivados del cambio de uso del suelo, y de sus efectos sobre la salud y el ambiente. Con éste proyecto se espera desarrollar herramientas metodológicas que permitan ponderar de manera objetiva y sistemática los factores de riesgo ambiental, obtener un perfil epidemiológico de la salud de las poblaciones estudiadas que sea representativo de la región, e interpretar los resultados obtenidos de manera interdisciplinaria. El conocimiento de la situación ambiental permitirá diseñar estrategias de acción que contribuyan a preservar la calidad del ambiente, mejorar los mecanismos de protección frente al uso de agroquímicos, proponer alternativas de producción agrícola ambientalmente sustentables, y colaborar al diseño de políticas públicas en materia de salud y ambiente. La intervención en el tejido social de las poblaciones a través de campañas de concientización, talleres de formación y el perfeccionamiento de la legislación vigente redundará en el cuidado del ambiente, y contribuirá a revertir algunos de los factores que inciden negativamente sobre el cambio climático. Se propone: 1. identificar factores de riesgo ambiental en agua, aire, sedimentos mediante la determinación de parámetros físicos, químicos, bioquímicos y biológicos apropiados. 2. fectuar un biomonitoreo genotóxico de personas expuestas laboralmente a plaguicidas, 3. efectuar un monitoreo de organismos bioindicadores potencialmente expuestos a plaguicidas en ambientes rurales, 4. establecer patrones de calidad ambiental 5. efectuar un estudio epidemiológico multivariado, georeferenciado, para obtener información estadística fehaciente sobre la situación socio ambiental de la región en relación a las nuevas prácticas productivas, 6. efectuar una devolución de los resultados obtenidos a través de informes periódicos a las autoridades 7. realizar talleres de formación en el seno de las comunidades orientados a fortalecer la prevención primaria de la salud y el cuidado del ambiente, 8. construir un cuerpo normativo a partir de la legislación vigente en materia de gestión ambiental 9. promover alternativas productivas que mitiguen el impacto ambiental del actual modelo y recuperar formas productivas con sustentabilidad ambiental y social. El proyecto se desarrollará en localidades rurales del interior provincial, y será llevado a cabo por un equipo interdisciplinario de las Universidades Nacionales de Córdoba, Rio IV y Villa María trabajando en red con médicos de hospitales regionales y centros primarios de salud con la colaboración de profesionales que trabajan en organizaciones sociales locales, redes sociales y grupos de participación ciudadana pre-existentes. Los laboratorios participantes cuentan con el equipamiento y los recursos humanos necesarios para la consecución de los objetivos del proyecto.

Creencias de autoefcacia y emociones de logro en estudiantes universitarios. Vinculaciones con contextos de evaluación

El bajo rendimiento advertido en general en las aulas universitarias, sumado a la abulia y escasa motivación por aprender que parece caracterizar a muchos estudiantes, acentúan el desafío que enfrentan las universidades de intervenir para mejorar la calidad de la educación de los ciudadanos. Conocer más acerca de los recursos internos y externos con que cuentan nuestros estudiantes para aprender y analizar el modo en que repercuten en su motivación y aprendizaje es, sin duda, un modo de avanzar hacia contribuciones capaces de redundar en mejoras educativas. En este marco, proponemos brindar aportes teóricos y definir líneas de acción que contribuyan al diseño de contextos de evaluación óptimos para el surgimiento de creencias motivacionales y estados emocionales beneficiosos para los aprendizajes de estudiantes universitarios. Suponemos que un contexto de evaluación que propicie el uso del conocimiento por parte de los estudiantes, que favorezca el despliegue de estrategias de autorregulación y posibilite ricos procesos de feedback, favorecerá la emergencia de percepciones de autoeficacia positivas, de estados emocionales beneficiosos y la ampliación de las posibilidades de lograr éxito en los resultados obtenidos. Trabajaremos con estudiantes de la Universidad Nacional de Río Cuarto, con muestras accidentales compuestas por la totalidad de alumnos que cursen Tráfico de señales –asignatura de Ing. en Telecomunicaciones- y Psicología Educacional -materia de distintos profesorados de la Facultad de Ciencias Humanas y de la Facultad de Ciencias Exactas Físico-Químicas y Naturales. Los datos serán recabados mediante el Multidimensional Scales of Perceived Self-efficacy (Bandura, 1990); el Achievement Emotions Questionnaire (Pekrun et al., 2005); análisis de protocolos de evaluación, observación no participante de clases e instancias de evaluación; grabación en audio de las clases observadas y entrevistas semi-estructuradas a estudiantes. Los resultados obtenidos prometen aportes teóricos y orientaciones pedagógicas. Las contribuciones teóricas se vinculan con la profundización de los marcos teóricos y con los avances logrados en las interpretaciones realizadas. Las orientaciones pedagógicas, se vinculan con aportes al diseño de contextos de evaluación óptimos para el surgimiento de creencias motivacionales y estados emocionales beneficiosos para los aprendizajes. Este proyecto se estima importante y pertinente porque materializa uno de los desafíos que enfrentan actualmente los investigadores educacionales, esto es, conocer más acerca del modo en que los aspectos personales de los estudiantes interactúan con las características del contexto académico en situaciones genuinas de aprendizaje; porque considera un tópico de investigación que emerge con fuerza en la agenda de la Psicología Educacional, esto es, el estudio de las emociones académicas; porque integra tendencias metodológicas que marcan rumbos en el estudio de las variables consideradas.

Entwicklungsprojekt MCUI13 : Machbarkeitsstudie/ Konzept zum Bau und Betrieb einer „Power to Methanol“-Anlage

[s.c.]

Action of growth regulators on soybean cultivar Davis

The effects of growth regularots on soybean plant (Glycine max) under greenhouse conditions were studied. Before flower ing, Agrostemmin (1 g/10 ml/3 1), gibberellic acid (GA) 100 ppm, and (2-chloroethyl) trimethylammonium chloride (CCC) 2,000 ppm were applied. At the flower anthesis, 2,3,5 - triio dobenzoic acid (TIBA) 20 ppm was applied. Other two applications with TIBA, with intervals of four days, were realized. Treatment with GA increased plant height while CCC presented a tendency to reduce it. Numbers of leaves, internods, and stems were not affected by the growth regulators.