Tendo em vista o notável crescimento da aquicultura mundial e principalmente a brasileira, torna-se cada vez mais importante o desenvolvimento de tecnologias que favoreçam esta prática. No entanto, a intensificação da piscicultura faz aumentar as chances de ocorrência de epizootias nos sistemas de produção como resultado do maior estresse imposto aos animais (lotação, manejo, transporte etc). Este estudo determinou a eficácia na utilização de imunoestimulantes dietéticos (levamisol e vitamina E) e a farmacocinética do imunoestimulante levamisol em juvenis de surubim Pseudoplatystoma sp. O levamisol dietético não afetou as variáveis de desempenho, mas influenciou positivamente o sistema imunológico inespecífico do cachara, afetando a atividade da lisozima sérica, sendo o melhor resultado observado para a concentração de levamisol de 247 mg kg-1 de ração. No estudo com vitamina E, não foram registrados efeitos sobre as variáveis de desempenho; ainda, na dose dietética 166 mg kg-1 o acetato de DL-?-tocoferol promoveu aumento das globulinas séricas. O estudo farmacocinético revelou-se que o levamisol é eliminado do sangue do cachara em 12 horas, sendo a meia-vida de distribuição (t1/2?) de 0,10 horas, e para o intervalo de eliminação meia-vida (t1/2?) de 1,86 horas. / Continuous stress negatively influences the immune system of farmed fish, increasing diseases susceptibility. In such a context, preventive measures, such as the administration of immunostimulants, are better suited for diseases control than using drugs and chemicals as remedies. This study aimed to evaluate the effects of dietary immunostimulants (levamisole and vitamin E) as well as the pharmacokinetic study of levamisole in juvenile of Pseudoplatystoma sp. Dietary levamisole did not affect growth parameters. However, increased lysozyme activity and lysozyme concentration, the best effects were recorded for fish fed 247 mg levamisole per kg of feed. In the vitamin E assay, no differences in growth parameters were recorded. However, vitamin E levels positively influenced the innate immune system increasing serum globulins at dose of 166 mg kg-1 of DL-?-tocoferol acetate. Also, at pharmacokinetic study levamisole is shown to be completely eliminated from fish blood on a 12-h period, being the distribution half-life (t1/2?) of 0.10 hours, whilst for the elimination interval the half-life (t1/2?) was 1.86 hours.

Este trabalho descreve o projeto de um modulador Analógico-Digital (A/D) Sigma-Delta de 16 bits (98 dB de SNR) de baixo consumo em tecnologia CMOS para a aquisição de sinais de áudio. Para projetar o modulador foi utilizada a metodologia top down, a qual consiste em projetar desde o nível de sistema até os blocos básicos em nível de transistores. O sistema foi analizado e projetado utilizando equacões e modelos comportamentais para obter as especificações de cada bloco do modulador. Considerando um baixo consumo de potência foi escolhida a topologia CIFF (do inglês Chain of Integrator with FeedForward) de terceira ordem e quatro bits implementado com capacitores chaveados. O modulador projetado é composto por três integradores chaveados, um somador analógico, um weigthed DAC e um quantizador de quatro bits. A técnica de Chopper é incluida no modulador para diminuir o ruído Flicker na entrada do modulador. Os blocos de maior consumo dentro do modulador são as OTAs. Por esta razão eles são projetados utilizando a metodologia gm/ID reduzindo assim o consumo de potência. O projeto foi realizado na tecnologia IBM 0,18 \'mü\'m sendo utilizado o simulador spectre do Cadence. O modulador Sigma-Delta atinge um SNR de 98 dB para uma banda de 20 kHz e um consumo de potência de 2,4 mW para uma fonte de alimentação de 1,8 V. / This work describes the design of a 16 bits low power Sigma-Delta modulator (98 dB SNR) in a CMOS technology for the acquisition of audio signals. To design the modulator it was used the top-down methodology, which consists on the design from system level to the transistor-level basic blocks. The system was analyzed and designed using behavioral models and equations to obtain the specifications of each block of the modulator. Considering a low power consumption it was chosen a third-order four bits CIFF topology (Chain Integrator with feedforward) implemented with switched capacitors. The modulator is composed by three integrators, one analog adder, one weigthed DAC and one four bit quantizer. The Chopper technique is included in the modulator to reduce the Flicker noise at the input of the modulator. The blocks of higher consumption within the modulator are the OTAs. Hence, they was designed using the methodology gm/ID to reduce power consumption. It was designed on the 0.18 \'mü\'m IBM technology and using the Cadence Spectre simulator. The Sigma-Delta modulator achieves a SNR of 98 dB for a bandwidth of 20 kHz and a power consumption of 2.4 mW with a 1.8 V power supply.

Orientadores: Hugo Enrique Hernandez Figueroa , Júlio César Rodrigues Fernandes de Oliveira / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia Elétrica e de Computação / Made available in DSpace on 2018-08-26T03:17:30Z (GMT). No. of bitstreams: 1 Freitas_AlexandrePassos_M.pdf: 2935936 bytes, checksum: 2bf660b1a9d7a183bc552d3db9da1b97 (MD5) Previous issue date: 2014 / Resumo: O crescente aumento da demanda de tráfego de dados dos sistemas de comunicação ópticos em conjunto com a busca da integração e miniaturização cada vez maior dos componentes impulsionaram a fotônica integrada em silício como uma das tecnologias promissoras para a evolução das novas gerações de dispositivos ópticos. Esta tecnologia, além de possuir suas características de um alto contraste de índice de refração, capacidade de modulação óptica através de controle de temperatura ou por densidade de portadores, se utiliza da infra-estrutura de fabricação para a indústria de microeletrônica já desenvolvida nas últimas décadas. Neste cenário, este trabalho propõe o desenvolvimento de um modulador de fase fabricado com a tecnologia de fotônica integrada em silício para o formato de modulação DP-QPSK e que opere na banda C de comunicação óptica. Análises de simulações e experimentais foram realizadas para a validação do fluxo de desenvolvimento do circuito e de cada componentes utilizado individualmente / Abstract: The increasing demand for data in optical communication systems with a constant search for reduction of device dimensions boosted silicon photonics as a candidate technology to the following optical device generations. Besides having high refractive index contrast, modulation capabilities through thermal or by carrier density control, this technology takes advantage of the microelectronic infra-structure developed in the last decades to fabricate small optical components with high reliability. In this scenario, this dissertation proposes the design of a phase modulator in silicon photonic technology. This modulator is able to operate at C-band and make the DP-QPSK modulation. Simulation and experiment analysis were made to validate the design flow for the optical circuit and for each single component / Mestrado / Telecomunicações e Telemática / Mestre em Engenharia Elétrica

La surinfection par HDV/ HBV est la forme la plus grave d'hépatite virale chronique et affecte entre 15-20 millions de patients au niveau mondial. HDV n'est pas susceptible aux traitements anti-HBV et le taux de réponse à l'IFNα est <25%. Malgré une progression plus rapide de la maladie hépatique, la majorité des patients présente une suppression de la réplication du HBV. Les détails des interactions entre HDV, HBV et le système immunitaire inné des cellules infectées restent inconnus. Les objectifs de ces travaux de thèse ont été: i) l'étude de l'infection par HDV et son interaction avec la réponse innée cellulaire; ii) l'identification de nouvelles stratégies thérapeutiques anti-HDV; iii) l'exploration de l'interaction entre HDV et HBV. L'approche expérimentale a été basée sur l'infection de cellules dHepaRG, capables d´entretenir des cycles réplicatifs complets de HBV et HDV et ayant une réponse immunitaire innée physiologique. Nous avons observé que: i) l'infection par HDV est associée à un réplication forte dans un nombre limité de cellules, et à une induction de l'expression des ISGs; ii) le traitement des cellules infectées par HDV avec de l'IFNα ne conduit pas à une induction accrue des ISGs et a une faible activité antivirale. Quelques agonistes de PRR, notamment activant la voie NF-kB, induisent une forte diminution de la réplication de HDV; iii) malgré le faible nombre de cellules infectées, HDV et ses protéines induisent une diminution de la réplication de HBV. Ces travaux ouvrent des perspectives importantes concernant la caractérisation de la pathogénèse de l'hépatite delta et l'identification de nouvelles stratégies thérapeutiques immuno modulatrices / HDV/HBV superinfection is the most aggressive form of chronic viral hepatitis and is estimated to affect 15-20 million patients worldwide. HDV is not susceptible to available direct anti-HBV drugs and sustained response to IFNα therapy occurs in less than 1/4 of patients. Despite the faster progression of liver disease, most HDV/ HBV infected patients present a suppression of HBV replication. The details of the interactions between HDV, HBV and the host cell innate immune response remain largely unexplored and research efforts have been limited by the lack of infection models. The aims of this thesis work were: i) to study HDV infection and the interplay with the host innate immune response; ii) to identify novel therapeutic strategies for the inhibition of HDV; iii) to further explore HDV/ HBV interference. The experimental strategy was based on infection of dHepaRG cells, which are known to be permissive to both HBV and HDV full replicative cycles and to present physiological innate immune responses. We observed that: i) HDV infection is associated with a strong, yet transient replication, a potent induction of the expression of ISGs; ii) IFN-α treatment of HDVinfected cells does not induce a further increase of ISG expression and has a modest antiviral activity. Conversely, some PRR agonists, in particular those inducing the NFkB pathway, induce a strong decline in HDV replication; iii) despite the low number of coinfected cells, HDV as well as its encoded proteins exert a repressive effect on HBV replication. Our work opens an array of perspectives on the pathogenesis of hepatitis delta and the identification of novel immune modulatory therapeutic strategies

Aus der Einleitung: "In der Historie erforderte die komplette dreidimensionale Aufnahme eines Objektes mittels Streifenprojektion einen hohen Zeitaufwand aufgrund der Realisierung der zeitlichen Abfolge von Streifensequenzen mit analogen Mitteln, mehrere Sekunden sind der übliche Standard für eine 3D Aufnahme. Damit einher geht die Notwendigkeit des während der Aufnahmezeit ruhenden Objektes. Das bedeutet Einschränkungen für eine Vielzahl von Anwendungen insbesondere bei dynamischen Prozessen und lebenden Objekten. Neue Möglichkeiten eröffneten sich mit der Einführung digitaler Lichtmodulatoren."

The region between the infrared and microwave region in the electromagnetic spectrum, the Terahertz (THz) gap, provides an exciting opportunity for future wireless communications as this band has been under utilised. This doctoral work takes a two-pronged approach into closing the THz gap with low-dimensional materials. The first attempt addresses the need for a compact THz source that can operate at room temperature. The second approach addresses the need to build optical elements such as filters and modulators in the THz spectrum. Terahertz quantum cascade lasers (THz QCLs) are one of the most compact, powerful sources of coherent radiation that bridge the terahertz gap. However, their cryogenic requirements for operation limit the scope of the applications. This is because of the electron-electron scattering and heating of the 2-dimensional free electron gas which leads to significant optical phonon scattering of the hot electrons. Theoretical studies in laterally confined QCL structures have predicted enhanced lifetime of the upper state through suppression of the non-radiative intersubband relaxation of carriers, which leads to lower threshold, and higher temperature performance. Lithographically defined vertical nanopillar arrays with electrostatic radius less than tens of nm offer a possible route to achieve lateral confinement, which can be integrated into QCL structures. A typical gain medium in a QCL consists of at least 100 repeat periods, with a thickness of 6-14 micron. For practical implementation of the top-down approach, restrictions are imposed by aspect ratios that can be achieved in present dry-etching systems. Typically, for sub-200 nm radius pillars, the thickness ranges from 1-3.5 micron. It is therefore necessary to work with THz QCLs based on 3-4 quantum well active regions, so as to maximise the number of repeat periods (hence gain) within an ultra-thin active region. After an introductory chapter, Chapter 2 presents a theoretical treatise on the realistic electrostatic potential in a lithographically defined nanopillar by scaling from a single quantum well (resonant tunnelling diode) to a THz QCL. Chapter 2 also discusses, the effect of lateral confinement on the intersubband states and the plasmonic mode in a THz QCL. One of the key experimental challenges in scaling down from QCLs to quantum-dot cascade lasers is the electrical injection into the nanopillars. This involves insulation and planarisation of the high aspect-ratio nanopillar arrays. Furthermore, the choice of the planarising layer is critical since it determines the loss of any optical mode. This experimental challenge is solved in Chapter 3. Chapter 4 presents the electro-optic performance of low-repeat period QCLs with an active region thickness that is less than 3.5 micron. Another topic of recent interest in the THz optics community is plasmonics in graphene. This is because the bound electromagnetic modes (plasmons) are tightly confined to the surface and can also be tuned with carrier concentration. Plasmonic resonance at terahertz frequencies can be achieved by gating graphene grown via chemical vapour deposition (CVD) to a high carrier concentration. THz time domain spectroscopy of such gated monolayer graphene shows resonance features around 1.6 THz superimposed on the Drude-like frequency response of graphene which may be related to the inherent poly-crystallinity of CVD graphene. Chapter 5 discusses these results, as an understanding of these features is necessary for the development of future THz optical elements based on CVD graphene. Chapter 5 finally describes how the gate tunability of THz transmission through graphene can be exploited to indirectly modulate a THz QCL. Chapter 6 presents ideas from this doctoral work, which can be developed in future to address the issues of enhanced temperature performance of THz QCLs and to realise realistic THz devices based on graphene.

A citocinese bacteriana é controlada por diversas proteínas que se agrupam em um complexo chamado divisomo. O cerne do divisomo é constituído por FtsZ, uma proteína homóloga à tubulina eucariótica, que se auto-associa formando uma estrutura chamada anel Z. O anel Z serve como arcabouço e recruta diversas outras proteínas componentes do divisomo para o sítio onde o septo será sintetizado na célula. A formação do anel Z é modulada por proteínas que se ligam diretamente a FtsZ e regulam a sua auto-associação, tanto induzindo como inibindo a sua polimerização. Apesar de muitos destes moduladores de FtsZ já serem conhecidos, muito pouco se sabe sobre o mecanismo pelo qual eles controlam a estruturação do anel Z in vivo. O objetivo do presente trabalho foi estudar a interação entre FtsZ e um modulador de divisão, a proteína ZapA, da bactéria gram-positiva Bacillus subtilis. Para isso construímos uma biblioteca de mutantes de ftsZ por \"Error Prone PCR\", com aproximadamente 1 substituição por cópia de ftsZ e contendo um total de 1x105 clones. A partir dessa biblioteca, utilizamos duas triagens genéticas para identificar mutantes incapazes de interagir com ZapA. Na primeira estratégia, selecionamos 12 mutantes de FtsZ resistentes à superexpressão de uma forma tóxica de ZapA, que bloqueia a divisão, causando filamentação e morte das células. Surpreendentemente, apesar destes mutantes serem insensíveis ao efeito de ZapA, ensaios citológicos mostraram que nenhum deles perdeu a interação com ZapA. Como as mutações foram mapeadas nas vizinhanças do sítio catalítico e de polimerização de FtsZ, e como a maioria delas confere resistência cruzada aos efeitos de outros moduladores de FtsZ, suspeitamos que elas afetassem a estabilidade do polímero de FtsZ e, consequentemente, o comportamento do anel Z. Essas suspeitas foram confirmadas em ensaios de FRAP e cálculos de proporção de FtsZ no anel Z, indicando que os mutantes formam um anel Z mais estável que o normal. Como não obtivemos mutantes que perderam a interação com ZapA na primeira triagem, aplicamos a biblioteca em uma segunda estratégia de triagem genética, procurando um mutante de FtsZ que voltasse a interagir com um mutante de ZapA que não se liga mais a FtsZ (ZapAN62A). Esta estratégia de ganho de função identificou um candidato, FtsZE91V , que, tanto por critérios genéticos como citológicos, voltou a interagir com ZapAN62A. Apesar do mutante FtsZE91V mostrar-se capaz de restaurar a interação com ZapAN62A, ele não afetou a interação com ZapA selvagem, segundo nossos ensaios de microscopia de fluorescência e viabilidade. O mutante FtsZE91V, mapeia na hélice H3 de FtsZ. Esta hélice está exposta na superfície de FtsZ (compõe um dos lados da molécula de FtsZ) de uma maneira compatível com a idéia de que ela seria importante para interações laterais entre polímeros de FtsZ. Nossos resultados apontam, portanto, que a hélice H3 deve ser o sítio de interação para ZapA em FtsZ. / The bacterial cytokinesis is ruled by a number of proteins that constitute the divisome complex. FtsZ, a homologue of eukaryotic tubulin, is the main component of the divisome and self-associates in a structure named Z ring. The Z ring works as a scaffold and recruits the other components of divisome, establishing itself where the septum will be synthesized in the cell. Some of these proteins interact directly with FtsZ and control self-association, promoting polymerization or preventing it. Although there have been discovered many of FtsZ modulators, little is known about the mechanisms that control the formation of the Z ring in vivo. The aim of this work was study de interaction between FtsZ e one of its division modulators, ZapA protein, on Bacillus subtilis grampositive bacteria. We created a mutagenized ftsZ plasmid library by error prone PCR, which contained 1,0x105 transformants and exhibited a mutation rate of one substitution per ftsZ copy. The library was transformed into a modified Bacillus subtilis strain and we performed two genetic screenings to select cells with FtsZ mutants incapable of interacting with ZapA. In first strategy, we selected 12 resistant ftsZ mutants for a toxic ZapA overexpression, that blocked division and caused filamentation and cell death. Surprisingly, although these mutants were insensitive to ZapA effect, cytological assays showed that none of them lost interaction with ZapA. As the substitutions were mapped around the catalytic and interaction site of FtsZ structure and showed resistance to other modulators, we suspected that the mutations were affecting the polymer stability of FtsZ and, consequently, the behavior of Z ring. This hypothesis was confirmed by FRAP experiments and by calculations of FtsZ proportions in Z ring, pointing out that the mutants form more stable Z rings. As we didnt\' find mutants that lost their ZapA´s interaction, we applied our library in a second genetic screen, looking for mutants that return to interact with a ZapA mutant (ZapAN62A) that doesn´t bind to FtsZ anymore. This gain of function strategy identified one candidate, FtsZE91V, which returns to interact with ZapAN62A in our genetic and cytological assays. Although the mutant FtsZE91V showed itself capable to interact with ZapAN62A, that didn´t affect the interaction with wild type ZapA by our fluorescent microscopy and viability assays. The substitution E91V was mapped on H3 helix of FtsZ structure. This helix is exposed on FtsZ surfaces (on FtsZ´s lateral side), being compatible with the idea that lateral interaction is important in FtsZ polymers. So, we concluded that helix H3 is the binding site of ZapA in FtsZ.

La maladie d’Alzheimer (MA) est une pathologie neurodégénérative lente, progressive et irréversible. Elle est principalement caractérisée par deux lésions histopathologiques : les dégénérescences neurofibrillaires, causées par l’accumulation intraneuronale d'agrégats de protéines Tau hyper- et anormalement phosphorylées ; et les dépôts amyloïdes parenchymateux, constitués d’agrégats de peptides amyloïdes (Aβ) issus du clivage de la protéine précurseur du peptide amyloïde (APP). À ces deux lésions s'ajoute entre autres une dérégulation de l'homéostasie protéique et des systèmes de dégradation des protéines : protéasome, autophagie et voie endosome/lysosome qui sont les mécanismes principaux mis en jeu pour l'élimination des agrégats protéiques. Plusieurs études suggèrent que la surproduction et l'agrégation des peptides Aβ sont les causes principales de la MA. Ainsi, la réduction de leur production et/ou l’amélioration de leur clairance représentent des stratégies potentielles pour le développement de traitement contre la MA. Les objectifs de mes travaux de thèse furent d’étudier de façon plus approfondie l’implication respective des voies de dégradation de l’APP et de ses métabolites ; de déterminer le mode d’action biologique de dérivés chimiques de la chloroquine sur le métabolisme de l’APP en relation avec la Valosin-Containing protéine (VCP), supposée être la cible de ces molécules, et par la même occasion d’étudier la relation existante entre l’APP et VCP. Dans un premier temps, nous avons pu démontrer que l’APP et ses fragments carboxyterminaux (APP-CTFs) étaient principalement dégradés par deux voies : la g-sécrétase et la voie endolysosome. Ensuite, nous avons démontré que VCP régulait le trafic intracellulaire de l’APP. Enfin, nous avons découvert que l’action des molécules était indépendante de la protéine VCP et qu’il s’agissait de modulateurs indirects de l’activité β-sécrétase permettant ainsi de réduire la sécrétion de peptides Aβ. En conclusion, ces travaux ont permis de contribuer à une meilleure compréhension du métabolisme de l’APP et de ses processus de dégradation ainsi que de caractériser l’effet biologique de nouveaux inhibiteurs indirects de la β-sécrétase. / Alzheimer's disease (AD) is a slow, progressive and irreversible neurodegenerative disease. There are two histopathological hallmarks found in AD brains: neurofibrillary tangles, caused by the intraneuronal accumulation of Tau protein aggregates in a hyper- and abnormally phosphorylated form; and amyloid deposits in the brain parenchyma which are mainly composed of amyloid peptides (Aβ) aggregates derived from the cleavage of the amyloid precursor protein (APP). AD physiopathology also includes a deregulation of protein homeostasis and degradation systems: proteasome, autophagy and the endosome/lysosome pathway, which are the main processes involved in the elimination of protein aggregates. Many evidences suggest that overproduction and aggregation of Aβ peptides are the main causes of AD and that strategies aiming to reduce their production and/or improve their clearance represent attractive approaches for AD therapeutics. Thus, this thesis aimed to study the respective contribution of g-secretase, proteasome and lysosomes in APP degradation; to determine the biological mode of action of chloroquine derivatives on APP metabolism in relationship with the Valosin-Containing Protein (VCP), supposed to be the target of these molecules, and at the same time, to study the relationship between APP and VCP. First, we have demonstrated that APP and its carboxy-terminal fragments (APP-CTFs) were mainly degraded by two pathways: g-secretase and the endosome/lysosome pathway. Next, we showed that VCP was involved in APP trafficking and processing. Finally, we have discovered that the action of our molecules does not depend on VCP but that they are indirect modulators of the β-secretase activity, reducing Aβ peptides secretion. In conclusion, this work contributed to a better understanding of APP metabolism and its degradation processes but also to characterize the biological effects of new indirect β-secretase inhibitors.

Aus der Einleitung: "In der Historie erforderte die komplette dreidimensionale Aufnahme eines Objektes mittels Streifenprojektion einen hohen Zeitaufwand aufgrund der Realisierung der zeitlichen Abfolge von Streifensequenzen mit analogen Mitteln, mehrere Sekunden sind der übliche Standard für eine 3D Aufnahme. Damit einher geht die Notwendigkeit des während der Aufnahmezeit ruhenden Objektes. Das bedeutet Einschränkungen für eine Vielzahl von Anwendungen insbesondere bei dynamischen Prozessen und lebenden Objekten. Neue Möglichkeiten eröffneten sich mit der Einführung digitaler Lichtmodulatoren."

Optogenetic approaches allow the activation or inhibition of genetically prescribed populations of neurons by light. In principle, optogenetics offers not only the ability to elucidate the functions of neural circuitry, but also new approaches to a treatment of neurodegenerative diseases and recovery of vision and auditory perception. Optogenetics already has revolutionized research in neuroscience. However, new methods for delivering light to three-dimensionally distributed structures e.g. in the brain are necessary. A major hurdle for focusing light through biological tissue is the occurring scattering and scrambling of the light. We demonstrate the correction of the scrambling in a multimode fiber by digital optical phase conjugation with a perspective for ptogenetics.