Le taux de mortalité dû au cancer est en hausse d’année en année. Le cancer du pancréas est l’un des plus mortels. Avec un taux de survie inférieur à 20% un an suivant le diagnostic, il y a une urgence pour développer de nouvelles molécules pour cibler cette maladie. La metformine, un biguanide utilisé cliniquement en tant qu’agent antidiabétique, s’est avérée à avoir des propriétés anticancéreuses. Les patients souffrant de diabète de type 2 prenant la metformine comme traitement sont moins à risque de développer plusieurs cancers dont celui du pancréas. Cependant, la metformine n’étant pas biodisponible, les doses à administrer seraient trop élevées pour la considérer comme thérapie anticancéreuse. Le groupe de recherche Schmitzer a synthétisé de nouveaux analogues de type biguanide plus lipophiles dans le but d’améliorer leur biodisponibilité. Le phényléthynylbenzyle biguanide (PEBB) est un des analogues présentant des propriétés antiprolifératives environ 800 fois plus puissantes que la metformine contre des cellules du cancer du pancréas. L’hexylbiguanide s’est aussi démarqué par sa spécificité pour les cellules cancéreuses et sa faible toxicité pour les cellules saines. Étant de bons candidats, des études in vivo ont été faites sur des souris en leur administrant le PEBB et l’hexylbiguanide afin d’obtenir des informations sur l’absorption et la distribution des composés. Pour ce faire, une méthode par LC-MS en mode multiple reaction monitoring (MRM) a été développée afin de quantifier différents analogues de biguanides dans le plasma de souris. De plus, une méthode par MALDI-TOF a été développée afin de localiser et quantifier les analogues dans les tissus par imagerie couplée à la spectrométrie de masse (IMS). Les expériences réalisées ont permis de suivre les composés dans le plasma et d’établir une cinétique d’absorption révélant que le PEBB atteint sa concentration plasmatique maximale environ à 1h après l’administration et que le composé est éliminé de la circulation sanguine à 80% au bout de 4h. Dans le cas de l’hexylbiguanide, la concentration plasmatique maximale est atteinte environ 30 minutes après l’administration pour être éliminé à plus de 90% après 4h. De plus, les études d’IMS ont révélé que le PEBB se distribue principalement dans le foie et légèrement dans les tumeurs. Aucune accumulation à long terme dans le foie n’a été observée, ce qui signifie que les risques de dommages hépatiques sont faibles. Les deux méthodes développées sont des méthodes puissantes IV et reproductibles afin de quantifier les différents types de biguanides dans les liquides biologiques ainsi que dans les tissus. / The death rate of cancer is increasing every year. Pancreatic cancer is one of the deadliest. With a survival rate of less than 20% one year post diagnosis, there is an emergency to develop new molecules to target this disease. Metformin, a biguanide clinically used as an antidiabetic agent, has been shown to have anticancer properties as well. Patients with type 2 diabetes taking metformin are less likely to develop several cancers including pancreatic cancer. However, due to the poor bioavailability of metformin, the doses would be too high to consider it as an anticancer treatment. The Schmitzer group has synthesized new biguanide analogues that are more lipophilic and thus more bioavailable. Phenylethynylbenzyl biguanide (PEBB) is one of the analogues with about 800 times more effective antiproliferative properties than metformin against pancreatic cancer cells. Hexylbiguanide also stood out for its specificity for cancer cells and its low toxicity for normal cells. In vivo studies were performed on mice by administering PEBB and hexylbiguanide to study the absorption and distribution of the compounds. For this aim, a LC-MS method was developed using Multiple Reaction Monitoring (MRM) mode to quantify different biguanide analogues in mice plasma. Complementarily, a MALDI-TOF method was developed to localize and quantify the analogues in tissues by imaging coupled to mass spectrometry (IMS). The experiments performed allowed to follow the compounds in plasma to establish absorption kinetics. These experiments revealed that PEBB reaches its maximum plasma concentration at 1h after administration and the compound is eliminated from the bloodstream at 80% after 6h. For hexylbiguanide, the maximum plasma concentration is reached about 30 minutes after administration and more than 90% is eliminated after 4 hours. In addition, IMS studies have shown that PEBB is distributed mainly in the liver and slightly in tumors. No accumulation in the liver was observed, which suggests that the risk of liver damage is low. These two methods are powerful and reproducible methods to quantify the different types of biguanides in biological fluids and tissues.

Avec l’omniprésence au quotidien du numérique et de l’informatique, de plus en plus d’utilisateurs souhaitent avoir accès à Internet et à leurs applications via n’importe quel périphérique, de n’importe où et n’importe quand. Les appareils domestiques intelligents se développant, les besoins d’échanger des données au domicile même se font de plus en plus sentir. C’est dans ce contexte, celui des services à domicile avec besoin d’interconnexion que se situe notre étude. Ce type de service est qualifié de Home Service (HS) alors que le réseau à domicile est nommé Home Network (HN). La problématique pour les opérateurs est alors de concevoir des architectures appropriées à l’interconnexion des HN de manière sécurisée tout en permettant un déploiement facile et à grande échelle. Dans la première étape, nous considérons la livraison de services sécurisés à travers un réseau de nouvelle génération (NGN) : IMS (IP Multimedia Subsystem). IMS étant l’architecture de référence pour son caractère réseau NGN des opérateurs, diverses architectures peuvent être développées comme support aux HS. Nous avons choisi d'analyser et de mettre en place une architecture P2P centralisée et de le comparer à l’architecture de référence. Plusieurs mécanismes d'authentification sont mis en place autour du P2P centralisé afin de sécuriser la prestation de services. La modélisation et l’évaluation de notre proposition ont permis d’identifier sa relation à l’IMS mais aussi des problèmes inhérents aux solutions centralisées : la protection des données personnelles, l’impact de la taille sur réseau sur les performances, l’existence d’un point de faiblesse unique face aux attaques et la congestion au niveau du serveur centralisé. Par conséquent, nous nous sommes tournés vers les solutions distribuées pour résoudre ces problèmes. Dans la deuxième étape, nous considérons l’architecture P2P non-structurée, qualifiée de pur P2P. La cryptographie basée sur l'identité (IBC) est ajoutée au P2P pur afin d’authentifier les utilisateurs et de protéger leurs communications. Pour chacune des solutions une analyse du coût de signalisation est effectuée révélant une faiblesse en ce qui concerne l’étape de recherche. Dans un déploiement à grande échelle, le coût de cette phase est trop élevé. Aussi, nous examinons le P2P structuré basé sur les Dynamic Hash Tables, une autre solution distribuée. Cette architecture est étudiée par l'IETF en tant qu’une des dernières générations de P2P: REsource LOcation And Discovery (RELOAD) Base Protocol. Nous proposons son utilisation dans le cadre des HSs. Comme preuve du concept, cette solution a été implantée et déployée sur un petit réseau en utilisant TLS/SSL comme mécanisme de sécurité. Cette plateforme nous a permis d’étudier les délais et les coûts de cette solution. Pour terminer, un bilan est établi sur toutes les solutions proposées En outre, nous introduisons d’autres types de HS et leurs possibilités de déploiement futur. / With digital life enhancement, more users would like to get seamless Internet and information with any devices, at any time and from anywhere. More and more home devices need to exchange data or to control other devices. The type of services is labelled Home Service (HS) and it is deployed though a Home Network (HN). Some users need to use their HS outside their HN, some others need to interconnect other HN. Operators have to provide suitable network architectures to ensure this interconnection and to provide at the same time, scalability, remote access, easy deployment and security. Here is the topic of our work. In the fist step, we consider a practical illustration around the Next-Generation Network (NGN) and the secured services. It is the IMS (IP Multimedia Subsystem) approach for the management of services that is generally supported by the NGN network operators. However, various network operator architectures can be developed to support these services. An alternative way is the P2P architectures. We choose to analyze and implement a centralized P2P and we compare it with the IMS solution. Several authentication mechanisms are introduced to secure the centralized P2P. An evaluation of these architectures is conducted. Since the previous solutions present some issues due to their centralized feature, we consider distributed solutions in a second step. The non-structured P2P, called pure P2P, can also support HS. Identity Based Crytography (IBC) is added to these architectures in order to offer authentication and protection to user communications. The different solutions are compared through their signaling and transmission cost. The study shows that searching step in this architecture is really costly, facing a scalability problem. Thus, we propose to use a structured P2P (called Dynamic Hash Table) for delivering HS between HN. This type of architecture is studied by IETF with the REsource Location And Discovery (RELOAD) Base Protocol. This solution is implanted and deployed here to be a proof of the concept. This test-bed enables the study of delay and security overhead in a real system. Eventually, the presented solutions are recaptured in order to see their advantages/ disadvantages. In addition, we introduce other perspectives in terms of HSs and network interconnection.

Pour la première fois, une étude par spectroscopie infrarouge à transformée de Fourrier et par microsonde ionique des minéraux majeurs de la croûte inférieure et des péridotites mantelliques a été entreprise afin de mieux caractériser les mécanismes d’incorporation et les teneurs en eau de ces minéraux nominalement anhydres, et de déterminer leurs compostions isotopiques en H, O, et Li pour retracer les échanges latéraux et horizontaux de l’eau dans la lithosphère continentale profonde. Les résultats montrent que: (1) Les minéraux nominalement anhydres, comme les pyroxènes et plagioclase, dans les granulites de la croûte inférieure contiennent de l’eau en trace essentiellement sous forme hydroxyles et accessoirement sous forme moléculaire, avec des concentrations (exprimées en poids H2O) allant de 200 à 2330 ppm pour les clinopyroxènes, de 60 à 1875 ppm pour les orthopyroxènes, de 65 à 900 ppm pour les plagioclases. Les teneurs calculées pour chaque roche totale d’après sa composition minéralogique et la teneur en eau des minéraux varient de 155 à 1100 ppm. (2) Les teneurs en H2O des minéraux majeurs et en roche totale de la croûte continentale inférieure sont manifestement plus élevées que celles du manteau lithosphérique sous-jacent, suggérant des variations verticales de la quantité d’H2O dans la lithosphère continentale profonde. Un tel contraste peut affecter de façon notable le comportement rhéologique de la lithosphère continentale. (3) Les rapports isotopiques de l’oxygène des pyroxènes étudiés, et probablement les roches totales, exprimés en [delta]18OSMOW , vont de ~ 4,5 à 12,5‰. Ceci indique la contribution de matériaux recyclés de la croûte continentale durant la pétrogenèse des échantillons ayant un TM18O élevé. (4) Les minéraux de la granulites sont caractérisés par des valeurs de élevées TMD, avec des valeurs de -80~-10‰ exprimées en [delta]DSMOW; les minéraux sont le plus souvent en équilibre les uns avec les autres lorsque l’on considère leurs compositions isotopiques moyennes. (5) Les compositions isotopiques du Lithium mesurée dans les minéraux de nos échantillons de granulites, exprimées en [delta]7Li par rapport à Lsvec, varient de -13 à +4.7 ‰. Ces valeurs sont donc pour la plus part inférieures à celles mesurées sur les MORB (2 – 6‰). La dispersion des valeurs reflètent l’hétérogénéité de la source des granulites, et les valeurs bassent resultent probablement de la perte par diffusion de Li pendant la mise en place des liquides silicatées provenant du manteau dans la croûte inférieure. (6) La grande hétérogénéité des teneurs en eau et en Lithium, et des compositions isotopiques de H-O-Li indique l’absence de circulation de fluide pervasive au travers de la croûte inférieure, qui aurait pour effet de supprimer les hétérogénéités à petite échelle et de les diminuer fortement à grande échelle / For the first time, systematic investigations of water content and H-O-Li isotopic compositions of minerals in lower crustal granulites, as well as water content of minerals in mantle peridotites, from eastern China have been carried out by Fourier transform infrared spectrometer and ion microprobe. The results show that: (1) Nominally anhydrous minerals, such as pyroxenes and plagioclase, in the lower crust generally contain trace amounts of structural water, with their content (H2O by wt.) varying from 200 to 2330 ppm for clinopyroxene, 60 to 1875 ppm for orthopyroxene, 65 to 900 ppm for plagioclase and 155 to 1100 ppm for the estimated bulk compositions. (2) Water contents of minerals in lower crustal granulites from eastern China, and their bulk values, are significantly higher than those in the underlying upper mantle, implying vertical heterogeneities of water distribution in the deep continental lithosphere; the contrast in water content even affect the rheological strength of the lithosphere. (3) The O-isotopic compositions of pyroxenes in the lower crustal granulites from eastern China are highly variable between different localities (~ 4.5 to 12.5‰, expressed in [delta]18OSMOW values), indicating variable influences from recycled crustal materials on their protoliths. (4) The H-isotopic compositions of granulite minerals from eastern China, are mostly in the range of -80 to -10‰ expressed in [delta]DSMOW values, and these minerals are usually in equilibrium with their [delta]D values. The relatively high [delta]D of granulite minerals may be related with degassing loss of H during the genesis of granulites. (5) The Li-isotopic compositions of granulite minerals from eastern China are usually in the range of -13 to 4.7‰, mostly lower than those of MORB (2-6‰). They reflect the source heterogeneity and are probably results of high-T Li diffusion during the intrusion of their original melts into the preexisting lower crust. (6) The large dispersion of Li and water contents and of H-O-Li isotopic results indicate the absence of any pervasive fluids in the lower crust

Esta pesquisa avaliou o uso de ozônio para inativar cistos de Giardia spp. e oocistos de Cryptosporidium parvum presentes na água filtrada obtida após a utilização da tecnologia de ciclo completo com flotação (coagulação, floculação, flotação e filtração) em escala de bancada, empregando o cloreto de polialumínio – PAC – como coagulante. O método de floculação em carbonato de cálcio – FCCa, sem e com a etapa de separação imunomagnética – IMS – foi utilizado na quantificação dos protozoários. A recuperação nos ensaios de qualidade utilizando o kit Easyseed® de Giardia spp. foi de 8,4% ± 97,4% sem IMS e com IMS, com duas dissociações, o valor foi de 7,4% ± 39,7% e, com três dissociações, a recuperação alcançou 9,6% ± 34,7%, portanto, somente o ensaio com IMS e duas dissociações, não atendeu os padrões do Método 1623.1. A recuperação de Cryptosporidium parvum, obteve valor de 3,4% ± 100% em ensaio sem IMS e com IMS, o valor obtido foi de 1,0% ± 70,0% com duas dissociações e 1,8% ± 44,4% com três dissociações e nos três métodos apresentados, não houve conformidade com os critérios do Método 1623.1. Na etapa de desinfecção com ozônio, os ensaios realizados na Etapa 1, que se utilizou 5 mgO3L-1 e tempo de contato de 1 min sem IMS, as maiores inativações atingidas foram de 2,52 e 2,22 log para cistos de Giardia spp. e oocistos de Cryptosporidium parvum, respectivamente. Com o tempo de contato de 5 min, as maiores inativações foram de 2,52 e 2,92 log de cistos de Giardia spp. e oocistos de Cryptosporidium parvum, respectivamente. Na Etapa 2, com IMS, utilizando a mesma dosagem e tempo de contato de 1 min, obteve-se 2,27 e 0,21 log de inativação para cistos e oocistos, respectivamente. Comparando-se com o tempo de contato de 5 min, foram obtidos 2,9 e 2,3 log de inativação para cistos e oocistos, respectivamente. Na avaliação de custo, o método de FCCa sem IMS demonstrou ser o mais econômico em relação ao procedimento com IMS. A influência da inclusão da terceira dissociação ácida no método com IMS também foi analisada e este procedimento não resultou em diferenças estatísticas significativas nos resultados. / This research evaluated the use of ozone to inactivate Giardia spp. cysts and Cryptosporidium parvum oocysts present in the filtered water obtained after the use of the complete cycle of flotation technology (coagulation, flocculation, flotation and filtration) on a bench scale employing polyaluminium chloride - PAC as coagulant. The calcium carbonate flocculation method - FCCa, without and with immunomagnetic separation step - IMS has been used in the quantification of protozoa. The recovery in quality test using the kit Easyseed® for Giardia spp. was 8.4% ± 97.4% non-IMS and IMS with two dissociations, the value was 7.4% ±39.7% and with three dissociations, the recovery reached 9.6% ± 34,7%, so only the test with IMS and two dissociations, did not meet the standards of method 1623.1. Recovery of Cryptosporidium parvum obtained value of 3.4% ± 100% in non-IMS and IMS testing, the value obtained was 1.0% ± 70.0% with two dissociations and 1.8% ± 44.4% with three dissociations and the three methods presented, there was non-compliance with the criteria of Method 1623.1. In step disinfection with ozone, tests performed in Step 1 was used 5 mgO3L-1 and contact time of 1 min without IMS, the major inactivation achieved were 2.52 and 2.22 log for Giardia spp. cysts and Cryptosporidium parvum oocysts, respectively. With 5 minutes of contact time, the greater inactivation were 2.52 and 2.92 log for Giardia spp. cysts and Cryptosporidium parvum oocysts, respectively. In Step 2, with IMS, using the same dose and 1 min contact time, there was obtained 2.27 and 0.21 log inactivation for cysts and oocysts, respectively. Compared with the 5 min of contact time, were obtained 2.9 and 2.3 log inactivation for cysts and oocysts, respectively. In evaluating cost, the FCCa method without IMS proved to be the most economical in relation to the procedure with IMS. The influence of inclusion of the third acid dissociation method in IMS was also analyzed and this procedure did not result in statistically significant differences in the results.

Este trabalho teve como objetivo avaliar diversos métodos de detecção e recuperação de cistos de Giardia spp. e de oocistos de Cryptosporidium parvum em resíduos gerados no tratamento de águas de abastecimento com turbidez elevada tendo como padrão o Método 1623.1 da USEPA (2012 ). Para tanto, ensaios utilizando aparelho Jarteste (coagulação, floculação, decantação e filtração ) foram realizados utilizando o coagulante cloreto de polialumínio - PAC. Em todos os métodos avaliados foi utilizada a técnica de purificação por separação imunomagnética - IMS. A adaptação do método floculação em carbonato de cálcio FCCa elaborado por Vesey et al. (1993) e adaptado por Feng et al. (2011), repercutiu nos melhores resultados para a amostra de resíduo sedimentado, com recuperações de 68 ± 17 % para oocisto de C. parvum e de 42 ± 7 % para cisto de Giardia spp. Entretanto, as recuperações para a amostra de água de lavagem dos filtros - ALF foram inferiores à 1 %, não sendo possível determinar um método adequado. A presença dos patógenos indica que o reuso da ALF em ETA convencionais ou o descarte em mananciais sem um tratamento prévio, pode representar problemas de contaminação. A adaptação dos métodos de Boni de Oliveira (2012) e Keegan et al. (2008), também repercutiram em porcentagens de recuperação expressivas para a amostra de resíduo sedimentado, sendo de: 41 ± 35 % para oocisto de C. parvum e 11 ± 70 % para cisto de Giardia spp., e 38 ± 26 % para oocisto de C. parvum e 26 ± 13 % para cisto de Giardia spp., respectivamente. A análise estatística não resultou em diferença significativa entre estes dois métodos, entretanto, as elevadas recuperações indicam que estes métodos podem ser melhor avaliados em pesquisas futuras. / This dissertation addresses the evaluation of several methods for the detection of Giardia spp. cysts and Cryptosporidium parvum oocysts in wastes produced after a high-turbidity water treatment, according to Method 1623.1 from USEPA (2012). Coagulant polyaluminium chloride - PACl was used in jar test experiments (coagulation/flocculation, sedimentation and filtration ). The Immunomagnetic Separation - IMS technique was applied to all methods. The calcium carbonate flocculation (CCF) method, developed by Vesey et al. (1993) and adapted by Feng et al. (2011 ), was applied to sludge samples in this research and was the best method tested, with 68% ± 17 % and 42 % ± 7,00 % recoveries for C. parvum oocysts and Giardia spp. cysts, respectively. On the other hand, the percentage recovery of (oo)cysts for filter backwash water samples was lower than 1 % and no suitable method could be detected. The presence of pathogens represents contamination risks for water sources and the reuse of filter backwash water may be a problem to conventional water treatment plants. The application of Boni de Oliveira (2012) and Keegan et al. (2008) methods, adjusted to this study, also resulted in significant percentage recoveries for the sludge samples, with 41 ± 35 % for C. parvum oocyst and 11 ± 70% for cyst Giardia spp., and 38 ± 26% for oocyst C. parvum and 26 ± 13% for cyst Giardia spp., respectively. The statistical analysis showed no significant differences between the two methods, however, such high recoveries indicate they should be better evaluated in future research.

Esta pesquisa investiga a adequação da abordagem de Learning Design como fundamentação teórico-prática para a contextualização do design instrucional. Partindo da premissa de que o aprendizado eletrônico é um fenômeno multidimensional, a pesquisa analisa as dimensões pedagógica, semântica, tecnológica, do aluno e organizacional, que correspondem aos interesses das diferentes comunidades envolvidas - professores e especialistas em educação, pesquisadores, especialistas em tecnologia, alunos e gestores. Utilizando a abordagem qualitativa de pesquisa e a perspectiva teórico-descritiva, os fundamentos teórico-práticos do Learning Design são cotejados com o estudo de caso do STEA - Sistema Transversal de Ensino-Aprendizagem, desenvolvido no âmbito do Núcleo de Educação de Jovens e Adultos e Formação Permanente de Professores (NEA) da Faculdade de Educação da Universidade de São Paulo (FEUSP). O caso contempla todos os processos do sistema de aprendizado eletrônico - do planejamento, design, implementação, execução e avaliação, até a formação e a atualização continuada de professores em serviço, abrangendo também a produção de materiais didáticos e ações complementares de pesquisa acadêmica, sob a perspectiva contextualizada do design instrucional. A pesquisa resultou na confirmação do Learning Design como fundamentação teórica para o design instrucional contextualizado, destacando-se que sua implementação nas práticas pedagógicas nacionais depende do aprimoramento das ferramentas de autoria, instanciação e execução de atividades de aprendizagem, para incorporar características da Web 2.0 e possibilitar a difusão do E-learning 2.0 como inovação no aprendizado eletrônico. / This research investigates if Learning Design approach can be seen a theoreticalpractical framework for contextualized instructional design. Based on e-learning as a multidimensional phenomenon, the research analyzes the pedagogical, semantic, technological, student and organizational dimensions, in correspondence to the interests of different e-learning communities - teachers and educational specialists, researchers, technology specialists, students and managers. Using qualitative approach and theoretician-descriptive perspective, the theoretical and practical framework of Learning Design are confronted to STEA case study, a transversal system for teaching and learning, developed by Nucleus of Adult and Young Education (NEA) of the College of Education of the University of São Paulo (FEUSP). Case study contemplates all the processes involved in e-learning system - planning, design, implementation, execution and evaluation, teachers education in service, enclosing the production of didactic materials and complementary actions of academic research, under a contextual approach of design instrucional. The research confirms Learning Design as theoretical framework for contextualized instructional design and distinguishes that its implementation in national practices depends on the improvement of authorship, instantiation and run tools, in order to incorporate features of Web 2.0 and to make possible the diffusion of E-learning 2.0.

O objetivo deste trabalho foi avaliar o desempenho da remoção de cistos deGiardia spp. e oocistos de Cryptosporidium parvum, em águas de abastecimento com turbidez elevada, em experimentos em escala de bancada (coagulação, floculação, decantação e filtração). Para tanto, empregou-se o coagulante cloreto de polialumínio – PAC. O método de filtração em membranas foi adotado para a concentração de protozoários, seguido ou não da etapa de purificação por separação imunomagnética – IMS. Os métodos foram avaliados em experimentos de controle de qualidade analítica e o método sem IMS apresentou as seguintes porcentagens de recuperação, 80% ±16,32% para cistos de Giardia spp. e 5% ±10,00% para oocistos de C. parvum. O método com IMS apresentou 31,5%±7,55% de recuperação para cistos de Giardia spp. e 5,75%±3,20% de recuperação para oocistos de C. parvum. Os experimentos demonstraram que não houve melhora na remoção de ambos os protozoários na condição de maior dosagem de coagulante (65 mg.L-1 de PAC e pH 7,29). A condição de menor dosagem de coagulante (25 mg.L-1 de PAC e pH 6,76) apresentou um desempenho melhor, ao contrário de uma expectativa de que a maior dosagem de coagulante pudesse favorecer a remoção destes microrganismos. A condição de menor dosagem apresentou, na água filtrada, 50 e 75% de ausência de identificação de cistos de Giardia e oocistos de C. parvum, respectivamente. A condição de maior dosagem apresentou (oo)cistos na água filtrada de todas amostras analisadas. Estes resultados indicam a importância do controle da coagulação na remoção de protozoários. / The aim of this study was to evaluate the performance of Giardia spp. cysts and Cryptosporidium parvum oocysts removal in a bench scale experiment. The coagulant polyaluminium chloride – PACl was used in this research. The protozoa concentration tests were performed by applying the Membrane Filtration method, with and without Immunomagnetic Separation assay-IMS. The methods were evaluated using control experiments and the method without IMS had the following percentage recovery, 80% ± 16.32% and 5% ±10.00% for Giardia cysts and C. parvum oocysts, respectively. The method with IMS presented 31.5% ± 7.55% and 5.75% ± 3.20% of percentage recovery for Giardia cysts and C. parvum oocysts, respectively. Bench scale experimental results have clearly shown that there was no improvement in protozoa removal using the superior dosage of coagulant. The inferior dosage condition (25 mg.L-1 of PACl and pH 6,76) performed better, which was contrary to what was expected in which a superior dosage of coagulant could favour when removing microorganisms. The inferior dosage condition presented 50% and 75% of absence of Giardia cysts and C. parvum oocysts in the final water, respectively. The second coagulation condition (65 mg.L-1 of PACl and pH 7,29) presented protozoa (oo)cysts in the final water of all the samples examined. These results indicate the importance of coagulation control in protozoa removal.

Esta pesquisa investiga a adequação da abordagem de Learning Design como fundamentação teórico-prática para a contextualização do design instrucional. Partindo da premissa de que o aprendizado eletrônico é um fenômeno multidimensional, a pesquisa analisa as dimensões pedagógica, semântica, tecnológica, do aluno e organizacional, que correspondem aos interesses das diferentes comunidades envolvidas - professores e especialistas em educação, pesquisadores, especialistas em tecnologia, alunos e gestores. Utilizando a abordagem qualitativa de pesquisa e a perspectiva teórico-descritiva, os fundamentos teórico-práticos do Learning Design são cotejados com o estudo de caso do STEA - Sistema Transversal de Ensino-Aprendizagem, desenvolvido no âmbito do Núcleo de Educação de Jovens e Adultos e Formação Permanente de Professores (NEA) da Faculdade de Educação da Universidade de São Paulo (FEUSP). O caso contempla todos os processos do sistema de aprendizado eletrônico - do planejamento, design, implementação, execução e avaliação, até a formação e a atualização continuada de professores em serviço, abrangendo também a produção de materiais didáticos e ações complementares de pesquisa acadêmica, sob a perspectiva contextualizada do design instrucional. A pesquisa resultou na confirmação do Learning Design como fundamentação teórica para o design instrucional contextualizado, destacando-se que sua implementação nas práticas pedagógicas nacionais depende do aprimoramento das ferramentas de autoria, instanciação e execução de atividades de aprendizagem, para incorporar características da Web 2.0 e possibilitar a difusão do E-learning 2.0 como inovação no aprendizado eletrônico. / This research investigates if Learning Design approach can be seen a theoreticalpractical framework for contextualized instructional design. Based on e-learning as a multidimensional phenomenon, the research analyzes the pedagogical, semantic, technological, student and organizational dimensions, in correspondence to the interests of different e-learning communities - teachers and educational specialists, researchers, technology specialists, students and managers. Using qualitative approach and theoretician-descriptive perspective, the theoretical and practical framework of Learning Design are confronted to STEA case study, a transversal system for teaching and learning, developed by Nucleus of Adult and Young Education (NEA) of the College of Education of the University of São Paulo (FEUSP). Case study contemplates all the processes involved in e-learning system - planning, design, implementation, execution and evaluation, teachers education in service, enclosing the production of didactic materials and complementary actions of academic research, under a contextual approach of design instrucional. The research confirms Learning Design as theoretical framework for contextualized instructional design and distinguishes that its implementation in national practices depends on the improvement of authorship, instantiation and run tools, in order to incorporate features of Web 2.0 and to make possible the diffusion of E-learning 2.0.

Este trabalho teve como objetivo avaliar diversos métodos de detecção e recuperação de cistos de Giardia spp. e de oocistos de Cryptosporidium parvum em resíduos gerados no tratamento de águas de abastecimento com turbidez elevada tendo como padrão o Método 1623.1 da USEPA (2012 ). Para tanto, ensaios utilizando aparelho Jarteste (coagulação, floculação, decantação e filtração ) foram realizados utilizando o coagulante cloreto de polialumínio - PAC. Em todos os métodos avaliados foi utilizada a técnica de purificação por separação imunomagnética - IMS. A adaptação do método floculação em carbonato de cálcio FCCa elaborado por Vesey et al. (1993) e adaptado por Feng et al. (2011), repercutiu nos melhores resultados para a amostra de resíduo sedimentado, com recuperações de 68 ± 17 % para oocisto de C. parvum e de 42 ± 7 % para cisto de Giardia spp. Entretanto, as recuperações para a amostra de água de lavagem dos filtros - ALF foram inferiores à 1 %, não sendo possível determinar um método adequado. A presença dos patógenos indica que o reuso da ALF em ETA convencionais ou o descarte em mananciais sem um tratamento prévio, pode representar problemas de contaminação. A adaptação dos métodos de Boni de Oliveira (2012) e Keegan et al. (2008), também repercutiram em porcentagens de recuperação expressivas para a amostra de resíduo sedimentado, sendo de: 41 ± 35 % para oocisto de C. parvum e 11 ± 70 % para cisto de Giardia spp., e 38 ± 26 % para oocisto de C. parvum e 26 ± 13 % para cisto de Giardia spp., respectivamente. A análise estatística não resultou em diferença significativa entre estes dois métodos, entretanto, as elevadas recuperações indicam que estes métodos podem ser melhor avaliados em pesquisas futuras. / This dissertation addresses the evaluation of several methods for the detection of Giardia spp. cysts and Cryptosporidium parvum oocysts in wastes produced after a high-turbidity water treatment, according to Method 1623.1 from USEPA (2012). Coagulant polyaluminium chloride - PACl was used in jar test experiments (coagulation/flocculation, sedimentation and filtration ). The Immunomagnetic Separation - IMS technique was applied to all methods. The calcium carbonate flocculation (CCF) method, developed by Vesey et al. (1993) and adapted by Feng et al. (2011 ), was applied to sludge samples in this research and was the best method tested, with 68% ± 17 % and 42 % ± 7,00 % recoveries for C. parvum oocysts and Giardia spp. cysts, respectively. On the other hand, the percentage recovery of (oo)cysts for filter backwash water samples was lower than 1 % and no suitable method could be detected. The presence of pathogens represents contamination risks for water sources and the reuse of filter backwash water may be a problem to conventional water treatment plants. The application of Boni de Oliveira (2012) and Keegan et al. (2008) methods, adjusted to this study, also resulted in significant percentage recoveries for the sludge samples, with 41 ± 35 % for C. parvum oocyst and 11 ± 70% for cyst Giardia spp., and 38 ± 26% for oocyst C. parvum and 26 ± 13% for cyst Giardia spp., respectively. The statistical analysis showed no significant differences between the two methods, however, such high recoveries indicate they should be better evaluated in future research.

Esta pesquisa avaliou o uso de ozônio para inativar cistos de Giardia spp. e oocistos de Cryptosporidium parvum presentes na água filtrada obtida após a utilização da tecnologia de ciclo completo com flotação (coagulação, floculação, flotação e filtração) em escala de bancada, empregando o cloreto de polialumínio – PAC – como coagulante. O método de floculação em carbonato de cálcio – FCCa, sem e com a etapa de separação imunomagnética – IMS – foi utilizado na quantificação dos protozoários. A recuperação nos ensaios de qualidade utilizando o kit Easyseed® de Giardia spp. foi de 8,4% ± 97,4% sem IMS e com IMS, com duas dissociações, o valor foi de 7,4% ± 39,7% e, com três dissociações, a recuperação alcançou 9,6% ± 34,7%, portanto, somente o ensaio com IMS e duas dissociações, não atendeu os padrões do Método 1623.1. A recuperação de Cryptosporidium parvum, obteve valor de 3,4% ± 100% em ensaio sem IMS e com IMS, o valor obtido foi de 1,0% ± 70,0% com duas dissociações e 1,8% ± 44,4% com três dissociações e nos três métodos apresentados, não houve conformidade com os critérios do Método 1623.1. Na etapa de desinfecção com ozônio, os ensaios realizados na Etapa 1, que se utilizou 5 mgO3L-1 e tempo de contato de 1 min sem IMS, as maiores inativações atingidas foram de 2,52 e 2,22 log para cistos de Giardia spp. e oocistos de Cryptosporidium parvum, respectivamente. Com o tempo de contato de 5 min, as maiores inativações foram de 2,52 e 2,92 log de cistos de Giardia spp. e oocistos de Cryptosporidium parvum, respectivamente. Na Etapa 2, com IMS, utilizando a mesma dosagem e tempo de contato de 1 min, obteve-se 2,27 e 0,21 log de inativação para cistos e oocistos, respectivamente. Comparando-se com o tempo de contato de 5 min, foram obtidos 2,9 e 2,3 log de inativação para cistos e oocistos, respectivamente. Na avaliação de custo, o método de FCCa sem IMS demonstrou ser o mais econômico em relação ao procedimento com IMS. A influência da inclusão da terceira dissociação ácida no método com IMS também foi analisada e este procedimento não resultou em diferenças estatísticas significativas nos resultados. / This research evaluated the use of ozone to inactivate Giardia spp. cysts and Cryptosporidium parvum oocysts present in the filtered water obtained after the use of the complete cycle of flotation technology (coagulation, flocculation, flotation and filtration) on a bench scale employing polyaluminium chloride - PAC as coagulant. The calcium carbonate flocculation method - FCCa, without and with immunomagnetic separation step - IMS has been used in the quantification of protozoa. The recovery in quality test using the kit Easyseed® for Giardia spp. was 8.4% ± 97.4% non-IMS and IMS with two dissociations, the value was 7.4% ±39.7% and with three dissociations, the recovery reached 9.6% ± 34,7%, so only the test with IMS and two dissociations, did not meet the standards of method 1623.1. Recovery of Cryptosporidium parvum obtained value of 3.4% ± 100% in non-IMS and IMS testing, the value obtained was 1.0% ± 70.0% with two dissociations and 1.8% ± 44.4% with three dissociations and the three methods presented, there was non-compliance with the criteria of Method 1623.1. In step disinfection with ozone, tests performed in Step 1 was used 5 mgO3L-1 and contact time of 1 min without IMS, the major inactivation achieved were 2.52 and 2.22 log for Giardia spp. cysts and Cryptosporidium parvum oocysts, respectively. With 5 minutes of contact time, the greater inactivation were 2.52 and 2.92 log for Giardia spp. cysts and Cryptosporidium parvum oocysts, respectively. In Step 2, with IMS, using the same dose and 1 min contact time, there was obtained 2.27 and 0.21 log inactivation for cysts and oocysts, respectively. Compared with the 5 min of contact time, were obtained 2.9 and 2.3 log inactivation for cysts and oocysts, respectively. In evaluating cost, the FCCa method without IMS proved to be the most economical in relation to the procedure with IMS. The influence of inclusion of the third acid dissociation method in IMS was also analyzed and this procedure did not result in statistically significant differences in the results.