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It is widely believed that we might be able produce life out of nonliving substances if we possessed the relevant knowledge. Thus synthetic biology is said to be on the way towards artificial life. But this is nonsense: “artificial life” cannot be thought. The idea that biological organisms could be produced reflects a misunderstandig of the concept “life”. Life is formally characterized by the fact that that which in the case of artifacts is three distinct activities – being something, producing it, and using it – is one. For a living being, to be is to be the source of its own activity through its own activity. Hence, if there is an activity of producing distinct from being what is produced – as would have to be the case in artificial life – what is thus produced is not life.

Various combinations of multi-stage thermophilic and/or mesophilic anaerobic sludge digestion systems were studied to evaluate their solids reduction, odor generation after centrifugal dewatering and indicator organism reduction in comparison to single-stage thermophilic and/or mesophilic anaerobic digestion systems. Pre-aeration of sludge in a thermophilic temperature was also tested followed by single or multi-stage anaerobic digestion systems. It was found that multi stage systems were capable of greater solids removal and placing thermophilic system in multi stage system enhanced indicator organism destruction below EPA Class A biosolids requirement. However, all the digestion systems in the study showed less than 3 log reduction of indicator organism DNA/g solids, which was much smaller than indicator organism reduction measured by standard culturing method. It was also found that the thermophilic anaerobic digestion system could increase organic sulfur-based odors from dewatered biosolids while placing a mesophilic digester reduced odors. It was exclusively observed from sludges containing high sulfate such as ones in this study. A combined anaerobic and aerobic sludge digestion system was also studied to evaluate their solids and nitrogen reduction efficiencies. The aerobic digester was continuously aerated to maintain dissolved oxygen level below 1 ppm and intermittently aerated. It was found that 90 % or more nitrogen removal was possible at the aerobic SRT greater than 3 days and the optimum aeration ratio could be determined. / Ph. D.

Cette thèse porte sur deux aspects complémentaires, le développement et l’implémentation de nouvelles approches de cristallisation et de cristallographie sérielle ainsi que leur mise en œuvre dans l’étude structurale de complexes enzymes : ARNt. La cristallographie est la méthode la plus employée en biologie structurale, mais elle présente encore des points délicats. Plusieurs méthodes avancées ont été déployées dans ce travail pour y pallier qui ont conduit à la résolution de la structure de l’ARNt nucléotidyltransférase du psychrophile Planococcus halocryophilus et à l’étude de son adaptation structurale au froid ; des puces microfluidiques de cristallisation qui ont servi à la résolution de plusieurs structures à température ambiante par cristallographie sérielle ; enfin le Xtal Controller utilisé pour l’étude d’évènements de nucléation et de croissance cristalline dans un but de préparation d’échantillons pour analyse sous rayonnement XFEL. Entre autres systèmes biologiques, cette thèse présente la caractérisation de deux familles d’inhibiteurs visant les aspartyl-ARNt synthétases, notamment du pathogène Pseudomonas aeruginosa. / This thesis focuses on two complementary aspects, the development and implementation of new approaches of crystallization and of serial crystallography as well as their use in the structural study of enzymes/tRNA complexes. Crystallography is the most used method in structural biology, but it presents delicate points. Different methods were implemented in this work to overcome these points, which led to the resolution of the structure of the CCA-adding enzyme of the psychrophilic organism Planococcus halocryophilus and to the study of its structural adaptation to the cold; novel microfluidic crystallization chips that have been used for the resolution of several structures by serial crystallography at room-temperature; finally the Xtal Controller used for the study of nucleation and crystal growth events with the purpose of preparing samples for analysis under XFEL radiation. Among other biological systems, this thesis presents the study and characterization of two families of inhibitors targeting aspartyl-tRNA synthetases, including the one of the pathogenic organism Pseudomonas aeruginosa.

Uppsatsen behandlar Sven-Harrys (Konstmuseum) utifrån ett poststrukturalistiskt perspektiv, där institutionens olika delar ställs i relation till varandra samt förankras i dess olika samhälleliga kontexter och ursprung. Institutionen Sven-Harrys jämförs med en organism, i syftet att förklara dessa relationer. Resultatet av undersökningen visar att Sven-Harrys verksamhet huvudsakligen syftar till att upplösa gränserna mellan konsten och bostaden genom att kombinera olika verksamheter som sammanknyter dessa.

Holography has the ability to render three-dimensional information of a recorded scene by capturing both the amplitude and phase of light incident on the recording medium. The application of digital camera technology and high-speed computing means digital holograms can be analysed numerically and novel applications can be found for this technology. This thesis explores the potential for both inline and off-axis digital holographic microscopy to study the three-dimensional swimming behaviour of bacteria. A high-magnification (225x) digital holographic microscope was designed and constructed with the ability to switch easily between inline and off-axis imaging modalities. Hardware aspects, in particular the illumination source, the choice of camera and data transfer rates, were considered. Novel strategies for off-axis holography combining dark field microscopy were designed and implemented. The localisation accuracy of the inline imaging modality was assessed by studying samples of polystyrene microspheres. The microscope is sensitive to stage drift on the order of angstroms per second and can successfully localise microspheres in dilute suspensions at least 100μm from the objective specimen plane. As a simple test of the capabilities of the microscope, the diffusion coefficient of a 0.5μm microsphere was found to be isotropic and consistent with the theoretical value. Amplitude and phase image reconstructions from the off-axis modality are demonstrated. High-magnification dark field off-axis holographic microscopy is shown to be superior to inline microscopy in localising 100nm gold nanoparticles. An artifact from our method of dark-field imaging, however, restricts the depth range to 15μm. A lower-magnification (45x) configuration of the microscope was used to study the 3D swimming behaviour of wild type Escherichia coli as a qualitative demonstration of the potential for this instrument in microbiological applications.

Nos últimos anos, a preparação de substâncias enantiomericamente puras tornou-se uma preocupação constante e um desafio para a indústria. Tal fato é consequência da relação tênue entre a estrutura espacial das substâncias e os diferentes sabores, odores, processos farmacocinéticos e farmacodinâmicos e toxicidade que elas desempenham no organismo. Nesse contexto, a síntese assimétrica de compostos carbonílicos ?-alquil substituídos opticamente ativos tem recebido grande atenção já que estes são importantes blocos de construção na síntese de fármacos quirais e produtos naturais. Assim, o presente trabalho buscou o desenvolvimento de metodologias que permitissem a preparação desses compostos enantiomericamente puros através do controle na formação de centros estereogênicos usando a biocatálise. Para isso, foram preparadas diversas olefinas ativadas por grupos retiradores de elétrons (carbonila) utilizando dois procedimentos, a saber, reações de condensação aldólica e esterificação. Esses compostos carbonílicos ?,?-insaturados foram obtidos com bons rendimentos (48-94%) e foram submetidos à reduções biocatalíticas mediadas por células íntegras de micro-organismos. Inicialmente, utilizou-se a olefina (E)-4-fenil-3-metil-3-buten-2-ona (51a) como substrato modelo para a otimização das condições experimentais e determinação daquela que fornecesse o produto biorreduzido com a melhor conversão e pureza enantiomérica. Nesses testes, avaliaram-se diversos parâmetros reacionais, tais como, composição do meio reacional (aquoso, bifásico e tamponante), presença de aditivos (glicose e catalisador de transferência de fase), diferentes micro-organismos e efeito do uso de resina adsorvente Amberlite XAD-7. Dessa forma, a biorredução em meio aquoso, sem aditivos, mediada pela levedura LSC-II e com a utilização da resina Amberlite XAD-7 foi a condição reacional que apresentou o melhor excesso enantiomérico (81%) e foi a selecionada para um estudo metodológico que avaliou os efeitos estérico e eletrônico dos substituintes. Observou-se que a posição do substituinte no ?-aril, o alongamento do ?-substituinte e a adição de um grupamento éster apresentaram um importante papel na estereosseletividade da reação. Por fim, avaliou-se a aplicação da metodologia na síntese enantiosseletiva da muguesia, uma fragrância floral. Os resultados obtidos revelaram o grande potencial da metodologia na biorredução quimiosseletiva de compostos carbonílicos ?,?-insaturados e preparação de substâncias opticamente ativas. / In recent years, the preparation of enantiomerically pure compounds has become a concern and a challenge to the industry. This fact is a consequence of the tenuous relationship between the spatial structure of substances and the different flavors, odors, pharmacokinetic and pharmacodynamic processes and toxicity that they play in the body. In this context, the asymmetric synthesis of ?-alkyl substituted optically active carbonyl compounds has received great attention because they are important building blocks in the synthesis of chiral drugs and natural products. Thus, this study aimed to develop methodologies that allow the preparation of this enantiomerically pure compounds by controlling the formation of stereogenic centers using biocatalysis. To this end, various activated olefins substituted with electron withdrawing groups (carbonyl) were synthesized using two procedures, aldol condensation and esterification reactions. These ?,?-unsaturated carbonyl compounds were obtained in good yields (48-94%) and subjected to the biocatalytic reduction mediated by whole cells of microorganisms. Initially, we used the olefin (E)-3-methyl-4-phenyl-3-buten-2-one (51a) as a model substrate for the optimization of the experimental conditions and determine wich one provide the product with better conversion and enantiomeric purity. In these tests, we evaluated various reaction parameters such as the composition of the reaction medium (aqueous, biphasic and buffer), the presence of additives (glucose and phase transfer catalyst), different microorganisms and adsorbing effect of using Amberlite XAD-7. Thus, the biorreduction in aqueous medium, without additives, mediated by LSC-II yeast and with the application of Amberlite XAD-7 was the reaction condition that presented the best enantiomeric excess (81%) and was selected for a methodological study that evaluated the steric and electronic effects of the substituents. It was observed that the substituent in the ?-position of the aryl group, the size increase of the ?-substituent and the presence of an ester group had an important role in the stereoselectivity of the reaction. Finally, application of the methodology on muguesia enantioselective synthesis, a floral fragrance, was evaluated. The results obtained showed the high potential of the methodology in chemoselective bioreduction of ?,?-unsaturated carbonyl compounds and preparation of optically active substances.

Com o advento da agricultura ampliou-se a produção alimentar e os bens de consumo, no entanto, os riscos ambientais também foram maximizados em função da adoção de técnicas produtivas baseadas no uso intensivo de insumos agrícolas. Esta problemática é mundial, embora mais evidenciada nos países em desenvolvimento e que tem, na produção agrícola, a base de sua economia. O Brasil enquadra-se nesta situação e desde 2009 é considerado o maior consumidor de agrotóxicos do mundo, criando um cenário de risco ambiental e de saúde humana. Os efeitos ambientais, base deste estudo, estão relacionados não somente à perda de espécies não-alvo, uma vez que os agrotóxicos não são seletivos, mas também as alterações em nível ecossistêmico, a qual se relaciona com as perdas das funções e dos serviços gerados pelos sistemas naturais. Adiciona-se a esta complexidade, a forma de ação de cada agrotóxico, a distribuição dos mesmos nos diferentes compartimentos (ar, solo e água), o período de permanência de cada um, as relações sinérgicas decorrentes das interações entre diferentes produtos, a formação de subprodutos no processo de degradação, entre outros fatores, como as diferenças existentes entre o ingrediente ativo e a formulação comercial, na qual existem os chamados ingredientes inertes em sua composição, os quais podem ser muito mais tóxicos para espécies e ecossistemas. Considerando esta abordagem, a presente pesquisa foi desenvolvida com base na realidade de um local de referência, o município de Bom Repouso (MG/BR), no qual a intensificação da produção de morango e batata tem trazido uma série de riscos sociais e ambientais. Semelhante a outras regiões produtivas do país, o uso de agrotóxicos é recorrente, amplo e irrestrito, com destaque para as formulações comerciais Kraft®36EC e Score®250EC, as quais, juntamente com seus respectivos ingredientes ativos (abamectina e difenoconazol), foram avaliadas por meio de testes de toxicidade com espécies de diferentes níveis tróficos representativas de um ecossistema aquático, gerando informações que foram avaliadas em nível de espécie e de ecossistema, simulando o cenário de aplicação dos produtos no local de referência. Os resultados obtidos permitiram concluir sobre as diferenças de sensibilidade das espécies e quais seriam as mais indicadas para se avaliar os efeitos tóxicos de ambos os agrotóxicos; os efeitos diferenciados entre a formulação comercial e os ingredientes ativos; bem como as respostas em termos de espécies e de ecossistemas, demonstrando a necessidade de que ambas as análises sejam consideradas na avaliação de risco ecológico. / With the advent of agriculture, food production and consumer goods has expanded, however, environmental risks were also maximized due to the adoption of production techniques based on intensive use of agricultural inputs. This worldwide problem is more evident in developing countries which have agricultural production as the basis of its economy. Brazil is into this situation and since 2009 it is considered the largest consumer of the world\'s pesticides, creating a scenario of environmental risk and human health. The environmental effects from this study are related not only to loss of non-targeted species, since pesticides are nonselective, but also changes in the ecosystem level, which relates to the loss of functions and services generated by natural systems. Add to this complexity, the mode of action of each pesticide, their distribution in the different compartments (air, soil and water), the period of stay of each, the synergistic relationships resulting from interactions between different products, the formation of by-products in the process of degradation, among other factors, such as differences between the active ingredient and the commercial formulation in which there are so-called ingredients inert in the composition, which may be more toxic to species and ecosystems. Considering this approach, this research was developed based on the reality of a reference site, the municipality of Bom Repouso (MG/BR), in which the intensification of production of strawberry and potato has brought a series of social and environmental risks. Similar to other productive regions of the country, the use of pesticides is recurrent, widespread and unrestrained, highlighting the commercial formulations Kraft®36EC and Score®250EC, which, along with their active ingredients (abamectin and difenoconazole) have been evaluated by toxicity tests with representative species of different trophic levels of an aquatic ecosystem, generating information that have been evaluated at the species level and ecosystem, simulating the application scenario of the products on the reference location. The results showed differences on the sensitivity of species and what are the most appropriate to evaluate the toxic effects of both pesticides; the differential effects between the commercial formulation and the active ingredients; as well as the responses in terms of species and ecosystems, demonstrating the need for both to be considered in ecological risk assessment.

Pour ce travail nous analysons le Pérou comme État d'origine et la France État d'accueil. Nous développons dans un Chapitre préliminaire l’importance de la Convention de La Haye du 29 mai 1993. En Première Partie Le chemin vers l’adoption d’un enfant péruvien par des Français, où existe deux conditions d’un côté établir que l'enfant péruvien est soumis à une vérification de sa situation légale dans le but de déclarer son adoptabilité et de l’autre qu'il y a des candidats français aptes et qualifiés évalués par des autorités compétentes françaises afin de déclarer leur qualification et leur aptitude à adopter. En Deuxième partie La réalisation de l’adoption au Pérou par des Français, le Secrétariat National d’Adoptions est l’autorité centrale chargée de s’assurer que les obligations assumées par l’Etat péruvien soient accomplies dans le cadre de la Convention de La Haye. Les adoptants français doivent transmettre leurs sollicitations au Secrétariat National des Adoptions qui va les apprécier et conclure l’adoption par résolution administrative. Puis en Troisième partie L’après adoption, les parents adoptifs doivent demander la reconnaissance par les autorités françaises de l’adoption réalisée au Pérou. L’adoption plénière provoque la rupture des liens avec la famille d’origine. Celle-ci n’empêchera pas les enfants adoptés s’ils atteignent l’âge de la majorité ou même s’ils sont encore mineurs de désirer connaître leur famille biologique. En guise de conclusion nous essayons de répondre à la question: L’adoption est-elle une véritable alternative à l’enfance abandonnée? / For this work we consider Peru as the state of origin and France as the receiving State. We develop in the preliminary Chapter the importance of the Hague’s Convention of 29 th May 1993. In the first Part: The way to the peruvian child adoption by french candidates where two conditions are considered: On one side the peruvian child who is subject to verification of his legal status with the intention to declare his adoptability, and on the other hand, the french candidates able to qualify, being evaluated by the french competent authorities in order to determine their suitability and their capability to adopt. In the Second Part: The accomplishment of the adoption in Peru by French Citizens, the Secretary National of Adoption is the central authority that makes sure that obligations assumed by the Peruvian State are enforced as part of the Hague’s Convention. The French candidates have to send their application to Secretary National of Adoption who, in turn will perform its function and, finally the adoption is completed by an administrative decision. After that in the Third part: After adoption, the adoptive parents have to demand the recognition of the French authority of the adoption realized in Peru. The plenary adoption causes the child to lose all ties with the biological family. This rupture does not prevent the adopted child from learning about his biological family, once he has reached adulthood. Conclussion: we set forth to answer the question: Is adoption a true alternative for abandoned children?