Das Influenza A Virus (IAV) stellt weltweit eine ernstzunehmende Bedrohung für Gesundheit und Wirtschaft der Menschheit dar. Ein universeller und langanhaltender Impfstoff konnte noch nicht entwickelt werden und klinisch zugelassene Medikamente verlieren durch die rasante Entstehung von resistenten Stämmen zunehmend ihre Wirkung. Aus diesem Grund gewinnt die Erforschung neuer antiviraler Strategien zur Bekämpfung des Influenzavirus an Bedeutung zum Schutze unserer Gesellschaft. Eine vielversprechende Zielstruktur für die Entwicklung neuer antiviraler Medikamente stellt das virale Hämagglutinin (HA) dar. Das HA liegt in hoher Dichte auf Influenzaviren vor und ermöglicht die Bindung an Sialinsäuren (SA) auf Wirtszellen und die Verschmelzung mit deren Lipidmembran. HA-bindende Moleküle entfalten eine hemmende Wirkung bereits bei dem ersten Kontakt mit Zellen, sodass eine Infektion erst gar nicht stattfinden kann. Aufgrund einer hohen HA-Dichte auf der Virusoberfläche eignen sich besonders multivalente SA tragende Nanopartikel für die Hemmung einer viralen Infektion. Aufbauend auf diesen Erkenntnissen, wurden in der vorliegenden Arbeit neue multivalente Binder gegenüber dem viralen Hämagglutinin (HA) entwickelt und studiert. Im Gegensatz zu bereits bekannten multivalenten Sialosiden, die in einer undefinierten räumlichen Orientierung auf Polymergerüsten präsentiert wurden, konnten in der vorliegenden Arbeit strukturelle Aspekte identifiziert werden, um Gerüstsysteme mit optimaler Rezeptorpräsentation gegenüber der Influenza A Virusoberfläche zu generieren. Neben SA-basierten Polymersystemen wurde auch ein gegen HA gerichtetes Peptid aus einem Antikörper identifiziert, welches sich auch für eine multivalente Interaktion mit IAV eignet. Diese Arbeit ermöglicht neue Einblicke in die Auswahl geeigneter Trägersysteme, eines optimalen Rezeptorabstandes und der Verwendung alternativer Rezeptoren mit dem Ziel einer Infektionshemmung von IAV. / Influenza A virus (IAV) still poses a serious threat to global health and economy of mankind. So far, a universal, long-lasting vaccine could not be developed, and clinically approved drugs are prone to lose activity due to the fast development of resistant strains. Because of this, research on new antiviral compounds and strategies to combat influenza viruses is of great importance for the protection of our society. A promising candidate for the development of novel antiviral drugs is the viral hemagglutinin (HA) protein. HA is present at high density on the viral envelope, which allows binding to sialic acid (SA) molecules on host cells and fusion with their membrane. Following, HA binding molecules have an inhibitory effect at the very first step of the infection cycle, leading to the inability of an infection. Based on a high HA density on the viral surface, SA carrying nanoparticles qualify for the inhibition of a viral infection. Based on this knowledge the study at hand demonstrates the development of new multivalent binders against viral HA and discusses them critically. In contrast to published multivalent sialosides, which are displayed in an undefined fashion on polymer scaffolds, the results of this thesis support the identification of structural requirements for the design of new scaffold systems with an optimal match to the viral surface. Beside sialoside based polymer systems, completely new peptide based systems, based on an HA binding antibody, were developed. Similar to polyglycerolsialosides, such multivalent peptide-decorated polymers were able to achieve nanomolar binding inhibition constants, too. In summary, this thesis enables new insights into the choice of a suitable carrier system, the optimal receptor spacing, and the use of alternative receptors with the ultimate goal of virus neutralization.

Micro RNA sind nicht-kodierende dsRNA ~22 Nukleotiden lang, die eine wichtige Rolle in der Entwicklung und Regulation in beinahe allen Eukaryoten spielen. MiRNAs binden target mRNA, was zu einer Blockierung der Proteintranslation führt. Viele Krankheiten sind bekannt, die durch veränderte miRNA Expressionsmuster entweder beeinflusst oder verursacht werden. Demnach könnte eine Manipulation der miRNA-Bildung einen therapeutischen Ansatz darstellen. MiRNA werden im Zytoplasma von längeren haarnadelförmigen prekursor RNA (pre-miRNA) durch das Enzym Dicer freigsetzt. Inhibition dieser Spaltung könnte durch spezifische pre-miRNA-bindende Moleküle erfolgen. Selektive Binder der pre-miRNA als Inhibitoren der miRNA-Reifung können durch testen von Substanzbibliotheken mit Hochdurchsatzscreening (HTS) gefunden werden. Diese Arbeit beschreibt den ersten homogenen Assay der miRNA-Reifung. Eine Fluoreszenzsonde in Form einer pre-miRNA wurde benutzt, die einen 5´-Fluorophor (FAM, Cy3, oder TMR) und einen 3´-Quencher (DABCYL) aufweist. Durch die nahe Nachbarschaft von Fluorophor und Quencher in der nativen Haarnadelstruktur erfolgt Fluoreszenzlöschung. Dicer spaltet diese Struktur effizient, was zur Dissoziation von Fluorophor und Quencher und somit zu einem Fluoreszenzanstieg führt. Der Assay wurde für HTS optimiert. Die ersten Verbindungen wurden auf deren Inhibition der miRNA-Reifung getestet. Mit einem Duplexassay, wobei zwei unterschiedliche pre-miRNA Sonden mit verschiedenen Fluorophoren eingesetzt wurden, konnte etwas spezifische Inhibition gezeigt werden. Der Assay wurde in Zellen durchgeführt und der Fluoreszenzanstieg mit Fluoreszenzmikroskopie detektiert. Somit ist ein Zell-basiertes Screening von Inhibitoren möglich. Eine einfachere Synthese der Sonde mittels in vitro Transkription und anschließender enzymatischen Ligation wurde entwickelt. Verwendung des Assays um hoch selective Inhibitoren der miRNA-Reifung zu entdecken könnte zu therapeutischen Ansätzen führen. / Micro RNAs (miRNAs) are non-coding dsRNAs of ~22 nucleotides that play a vital role in development and regulation in nearly all eukaryotes. MiRNAs bind target mRNA, thus blocking protein translation. Many diseases have been found to be influenced or caused by aberrant expression of miRNAs. A manipulation of miRNA formation may have therapeutic potential. MiRNAs are cleaved from longer hairpin precursor RNA (pre-miRNA) in the cytoplasm by the enzyme Dicer. It might be possible to inhibit this cleavage through specific pre-miRNA binding molecules. Selective binders of pre-miRNA as inhibitors of miRNA maturation can be found by testing large libraries of substances through high throughput screening (HTS) using an appropriate assay. This work describes the first homogenous assay of miRNA maturation. A fluorescent probe in the form of a pre-miRNA containing a 5´-fluorophore (FAM, Cy3, or TMR) and a 3´-quencher (DABCYL) was used. This ‘beacon’ in its native hairpin formation brings the fluorophore and quencher moieties into close proximity, resulting in fluorescence quenching. Dicer efficiently cleaves this structure, leading to dissociation of fluorophore and quencher and thus a fluorescence increase. In the presence of an RNA ligand that blocks cleavage, a lower fluorescence increase is seen. The assay was optimized for HTS. The first compounds were tested for their inhibition of miRNA maturation. Using a duplex assay, with two different pre-miRNA probes each containing a different fluorogenic group, some specific inhibition was shown. The assay was performed in cells using fluorescence microscopy to measure the fluorescence. This would allow for a cell-based screening of inhibitors. A simpler approach of beacon synthesis using in vitro transcription followed by enzymatic ligation was also established. Use of this assay to discover highly selective inhibitors of miRNA maturation may lead to disease therapeutics.

Die hydrolysestabile C-C-Bindung von Nukleosiden, deren Nukleobase über ein aromatisches oder methylen-verbrücktes Kohlenstoffatom an Ribose oder 2-Desoxyribose gebunden ist, ermöglicht die Synthese von neuartigen Strukturen und Eigenschaften, die bei N-Nukleosiden nicht stabil oder nicht gegeben wären. In dieser Arbeit wurde die die Cuprat-vermittelte Glycosylierung und die Friedel-Crafts-Alkylierung als Methoden zur Darstellung von Desoxyribose-basierenden C-Nukleosiden weiterentwickelt. Die Cuprat-vermittelte Glycosilierung ermöglichte die Synthese von C-Nukleosiden in bis zu 93% Ausbeute, wenn Grignard-basierende Normant-Cuprate verwendet wurden. Die Verwendung Organolithium-basierender Gilman-Cuprate war ebenfalls möglich. In Gegenwart von Sauerstoff wurden O-Glycoside isoliert in über 80 % Ausbeute isoliert. Mit den C-Nukleosiden wurden modifizierte Oligonukleotide, die als potentiell verbesserte Binder an M.TaqI und E.coli Dam dienen, dargestellt. Nach ihrer Charakterisierung über Schmelzwerte und Fluoreszenzeigenschaften wurde diese an die Arbeitsgruppe von Prof. Elmar Weinhold weitergereicht und dort erfolgreich als optimierte Binder an an M.TaqI und E.coli Dam getestet. Oligonukleotide, die ein oder mehrere 1,1-Binaphthyl-Chromophore als einen neuen Typus eines torsionsflexiblen Farbstoffes enthalten, wurden untersucht. Die Einführung mehrerer aufeinander folgender Binaphthyle führte zu einer thermodynamischen Stabilisierung von Duplex-Oligonukleotiden. Die geringe Neigung Binaphthyls zur Selbstlöschung bewirkte dabei einen starken Anstieg der Fluoreszenz. / The stable C-C-bond of ncleosides, whose nucleobase is attached to the ribose or 2-deoxyribose via an aromatic or methylen-bridged carbon atom, is stable to hydrolyses. This allows the synthesis of new structures and properties, which would not be available in N-nucleosides. In this work, a cuprate-mediated glycosilation and the Friedel-Crafts-alkylation as methods for the preparation of doxyribose-based C-nucleosides were developed. The cuprate-mediated glycosilation allowed the synthesis of C-nucleosides in up to 93 % yield, when Grignard-based Normant-Cuprates were used. The use of Organolithium-based Gilman-Cuprates was also possible. In the presence of oxygen O-glycosides were isolated in over 80 % yield. With the C-nucleosides modified oligonucleotides, which serve as potentially improved binders of the DNA-methyltransferases M.TaqI und E.coli, were prepared. After their charakterisation via melting point measurements and fluorescence properties, the oligonucleotides were given to the working group of Prof. Elmar Weinhold and successfully tested as improved binders of the DNA-methyltransferases M.TaqI und E.coli. Oligonucleotides, which contain one or multiple 1,1-binaphthyles as a new type of a torsionally flexible chromophore, were charakterised. The incorporation of several successive binaphthyls led to a thermodynamical stabilisation of the duplex-oligonucleotides. The low tendency of the Binaphthyl for self-quenching caused a remarkable increase of the fluorescence.

Trata-se de uma investigação sobre a influência de alguns fatores no comportamento mecânico de misturas asfálticas densas quanto à estabilidade e à flexibilidade. Foram testados três tipos de ligantes, asfalto convencional, modificado com 4,5% de SBS e modificado com 20% de borracha reciclada de pneu; duas distribuições granulométricas do agregado, centros das faixas B e C do DNER (1997); quatro teores de ligante, escolhidos com base nos valores de volumes de vazios e espessuras de película almejados; três condições de envelhecimento a longo prazo, mistura não envelhecida, envelhecida em estufa ventilada a 85ºC por 5 dias e envelhecida por exposição ao tempo por 4 meses; e duas condições de envelhecimento a curto prazo, mistura não envelhecida e envelhecida em estufa ventilada a 135ºC por 4 horas. A técnica de planejamento e análise de experimentos fatoriais fracionários assimétricos foi utilizada para a consideração simultânea dos fatores citados. O comportamento mecânico das misturas foi avaliado através dos ensaios de resistência à tração, módulo de resiliência e fluência por compressão uniaxial estática e dinâmica. A análise de variância dos resultados permitiu a identificação dos fatores com influência significativa nas respostas dos ensaios. O modo como cada fator interferiu nas propriedades apresentadas pelas misturas foi estabelecido através da construção de modelos estatísticos de comportamento. Os resultados mostraram que a adição de modificadores ao asfalto melhora a resistência das misturas à fadiga e à deformação permanente. Os processos de envelhecimento aumentaram os módulos de resiliência das misturas mas diminuíram sua capacidade de recuperação elástica, o que significa queda de resistência à fadiga. Por outro lado, a resistência a deformações permanentes das misturas, inclusive daquelas compostas por asfaltos modificados, aumentou com o envelhecimento. / This work deals with the influence of some factors in the mechanical behavior of asphalt mixtures stability and flexibility. Three types of binders were tested, conventional one, modified with 4,5% of SBS and modified with 20% of recycled tire rubber. Two aggregate gradations were tested, center of B and C gradations of DNER (1997). Four binder contents were chosen based on the air voids and film thickness. Three types of long-term aging were tested, not aged, aged in a forced-draft oven for 5 days at 85ºC and aged through weather exposition for the period of 4 months. Two types of short-term aging were tested; not aged and aged in a forced-draft oven for 4 hours at 135ºC. The technique of design and analysis of asymmetric fractional factorial experiments was used for the simultaneous analysis of the factors. The mechanical behavior of the mixtures was evaluated based on indirect tensile strength test, resilient modulus test and static and dynamic creep tests. The results of the analysis of variance allowed the identification of factors with significant influence in the answers. The influence of the factors in the mixtures properties was established through statistical models of behavior. The results showed that modified binders improves the mixtures resistance in relation to fatigue and to permanent deformation. The aging processes increased the resilient modulus of the mixtures, but also decreased its capacity of elastic return, resulting in a lost of fatigue resistance. On the other hand, the permanent deformation resistance of the aging mixtures increased, including the mixtures with modified binders cases.

O ensaio de fluência e recuperação sob tensão múltipla (MSCR) é resultado do aperfeiçoamento do ensaio de fluência repetida e recuperação (RCRT) pela Administração Rodoviária Federal dos Estados Unidos (FHWA) e tem por objetivo avaliar o percentual de recuperação, a compliância não-recuperável e a dependência dos ligantes asfálticos, especialmente os modificados, quanto ao nível de tensão. O MSCR foi utilizado neste estudo para caracterizar o comportamento fluência-recuperação de diversos ligantes asfálticos modificados virgens e envelhecidos a curto prazo. Foi empregado o procedimento da norma ASTM D7405, que prescreve os tempos de fluência e recuperação de 1 e 9 s, e avaliada a influência do aumento dos tempos de fluência e recuperação de 1 e 9 s para 2 e 18 s nas propriedades dos ligantes asfálticos envelhecidos a curto prazo. Além dos resultados destes ensaios, foram utilizados os valores de penetração, ponto de amolecimento, viscosidade rotacional, penetração retida, incremento do ponto de amolecimento, incremento de viscosidade e perda de massa para verificar quais ligantes asfálticos apresentam melhor comportamento à luz das propriedades e parâmetros considerados. Foram preparados doze materiais modificados e de classificação PG 76-XX tendo como base um CAP 50/70 de classificação PG 64-XX: CAP+PPA, CAP+Elvaloy+PPA, CAP+borracha, CAP+borracha+PPA, CAP+SBS, CAP+SBS+PPA, CAP+EVA, CAP+EVA+PPA, CAP+PE, CAP+PE+PPA, CAP+SBR e CAP+SBR+PPA. O CAP+Elvaloy+PPA apresenta, em um contexto geral, o melhor desempenho nas propriedades e parâmetros analisados em virtude dos bons resultados do percentual de recuperação e da compliância não-recuperável e das menores sensibilidades à tensão. O CAP+PE apresenta o pior desempenho geral dentre os materiais estudados, na medida em que os resultados do percentual de recuperação e da compliância não-recuperável deste ligante asfáltico são ruins e as sensibilidades ao aumento nos tempos de fluência e recuperação de 1 e 9 s para 2 e 18 s são elevadas. Ao ordenar os ligantes asfálticos do melhor para o pior à luz das propriedades e parâmetros considerados, o resultado é: CAP+Elvaloy+PPA, CAP+EVA, CAP+EVA+PPA, CAP+PPA, CAP+borracha+PPA, CAP+SBR+PPA, CAP+SBS+PPA, CAP+SBS, CAP+PE+PPA, CAP+borracha, CAP+SBR, CAP 50/70 e CAP+PE. / The multiple stress creep and recovery (MSCR) test is the result of a refinement of the repeated creep and recovery test (RCRT). This refinement was performed by the United States Federal Highway Administration (FHWA) with the aim of evaluating the percent recovery, the non-recoverable compliance and the stress dependency of asphalt binders, especially the modified ones. The MSCR test was applied in this study in order to characterize the creep-recovery behavior of unaged and short-term aged modified asphalt binders. The test procedure prescribed in ASTM D7405 specification was adopted and, to evaluate the influence of a greater creep-recovery time in the behavior of asphalt binders, the creep and recovery times of 2 and 18 s were also adopted in additional tests. The results of these tests, as well as the penetration, softening point, rotational viscosity, retained penetration, softening point increase, viscosity increase and mass loss data and mixing and compaction temperatures, were used with the objectives of ranking the asphalt binders and checking the ones with the best results in the parameters and properties considered. By using a PG 64-XX base asphalt-binder of 50/70 penetration grade, the following PG 76-XX odified asphalt binders were prepared: AC+PPA, AC+Elvaloy+PPA, AC+crumb rubber, AC+crumb rubber+PPA, AC+SBS, AC+SBS+PPA, AC+EVA, AC+EVA+PPA, AC+PE, AC+PE+PPA, AC+SBR and AC+SBR+PPA. In general, AC+Elvaloy+PPA has the best results in the parameters and properties considered in this study, especially due to the good results in the percent recovery and non-recoverable compliance and the lower stress sensitivity. In general, AC+PE has the worst results among the asphalt binders considered in this work, due to the facts that the percent recovery and non-recoverable compliance results are not good and the sensitivities to the creep-recovery times are high. By taking the results of this study and ranking the asphalt binders from the best to the worst, the following sequence is obtained: AC+Elvaloy+PPA, AC+EVA, AC+EVA+PPA, AC+PPA, AC+crumb rubber+PPA, AC+SBR+PPA, AC+SBS+PPA, AC+SBS, AC+PE+PPA, AC+crumb rubber, AC+SBR, neat AC and AC+PE.

O envelhecimento dos ligantes asfálticos possui uma grande influência sobre as propriedades mecânicas das misturas asfálticas. As especificações atuais limitam determinados índices ou parâmetros de envelhecimento, visando contornar o endurecimento excessivo do ligante asfáltico, porém, não levam em conta o efeito da radiação ultravioleta (UV). O objetivo deste trabalho é avaliar o efeito dos envelhecimentos termo-oxidativo e foto-oxidativo sobre as propriedades reológicas de um CAP puro e 11 ligantes asfálticos modificados, incluindo polímeros elastoméricos e plastoméricos, borracha moída de pneus e ácido polifosfórico, incluindo também combinações destes aditivos. Os ligantes asfálticos, com classificação por grau de desempenho PG 76-XX, preparados com base em um CAP 50-70 com classificação PG 64-22, foram previamente envelhecidos a curto prazo na estufa RTFO e posteriormente submetidos ao envelhecimento foto-oxidativo em uma estufa UV sob dois níveis de radiação, 0,68 W/m² e 1,00 W/m², por um período de 240 h. Para efeitos de comparação dos resultados, as mesmas amostras foram submetidas ao envelhecimento termo-oxidativo na estufa PAV. As amostras virgens e envelhecidas a curto prazo foram caracterizadas por meio de ensaios empíricos. Os resíduos obtidos nos envelhecimentos foram submetidos a ensaios no reômetro de cisalhamento dinâmico (DSR). Propriedades reológicas dos ligantes asfálticos foram analisadas por meio de curvas-mestre e do ensaio de fadiga LAS. Algumas amostras foram caracterizadas quimicamente por meio de espectroscopia no infravermelho (FTIR), cromatografia por permeação em gel (GPC) e do fracionamento SARA. Para a maioria das amostras os procedimentos PAV e UV a 0,68 W/m² apresentam certa equivalência, em termos da capacidade de alterar as propriedades reológicas analisadas. O ligante asfáltico que mostrou ser menos sensível ao envelhecimento PAV foi o CAP+SBR, o que demonstrou ser menos sensível ao envelhecimento UV a 0,68 W/m² foi o CAP+EVA, e o que demonstrou menor sensibilidade ao envelhecimento UV a 1,00 W/m² foi o CAP+SBS. O CAP+SBS+PPA foi a amostra que apresentou maior sensibilidade ao envelhecimento PAV e UV a 1,00 W/m². / Aging of the asphalt binders has a great influence on the mechanical properties of asphalt mixtures. The current specifications establish limits for some indexes or aging parameters in an attempt to avoid the excessive hardening of the asphalt binder; however, they do not take into account the effects of ultraviolet (UV) irradiation. The objective of this study is to analyze the impact of thermo-oxidative and UV aging methods on the rheological properties of a base asphalt cement (AC) and 11 formulations prepared with additives, i. e., elastomeric and plastomeric polymers, ground tire rubber and polyphosphoric acid. Combinations these additives were also included in the study. The modified asphalt binders are graded as PG-76 XX in the Superpave specification, whereas the 50-70 base AC is graded as PG 64-22 in the same specification. The materials were first aged in the rolling thin-film oven (RTFO), and then submitted to photo-oxidative aging in an UV oven at two irradiation levels (0.68 W/m² and 1.00 W/m²) for 240 h. For comparison purposes, these formulations were also subjected to thermo-oxidative aging in the pressurized aging vessel (PAV). The unaged and short-term aged samples were characterized by means of empirical tests. The residues obtained in the RTFOT, PAV e UV aging methods were tested in the dynamic shear rheometer (DSR). The rheological properties of asphalt binders were analyzed by means of master curves and the linear amplitude sweep (LAS) fatigue test. Some samples were chemically characterized by the Fourier transform infrared spectroscopy (FTIR), the gel permeation chromatography (GPC) and the SARA fractionation techniques. The PAV and the UV procedures at 0.68 W/m² show some similarities for many formulations in terms of the ability to change their rheological properties. The AC+SBR was found to be less sensitive to PAV aging than the other studied binders, whereas the AC+EVA was the less sensitive material to UV aging at 0.68 W/m² and the AC+SBS showed the lowest sensitivity to UV aging at 1.00 W/m². The AC+SBS+PPA was the most sensitive formulation to PAV and UV aging at 1.00 W/m².

A adição de modificadores no ligante asfáltico visando melhorar o seu comportamento é uma prática comum atualmente. Existem vários tipos de modificadores, entre eles o mais utilizado nas rodovias federais brasileiras é o copolímero SBS. Além dos polímeros, o ácido polifosfórico (PPA) é outro tipo de modificador que produz boas características quando adicionado ao ligante asfáltico. Apesar do efeito positivo, sabe-se que este material é importado, o que dificulta sua aquisição e pode tornar o produto final muito caro. Tendo em vista a possibilidade de se trabalhar com um produto de mesma base, mas de origem nacional, o que o tornaria o modificador mais acessível, optou-se por verificar o comportamento da adição de ácido fosfórico (PA85) ao ligante asfáltico. Sendo assim, o propósito desta tese é analisar o efeito da adição de ácidos fosfóricos no comportamento reológico de ligantes asfálticos puros e modificados com o copolímero SBS. Para tanto, realizou-se ensaios empíricos e reológicos com o intuito de verificar o comportamento destes materiais. Na primeira parte da pesquisa, dois ligantes asfálticos de base, CAP50/70 de classificação PG64-XX, (Replan e Lubnor), foram modificados somente com a adição dos dois ácidos (PPA e PA85), enquanto na segunda parte esses ligantes asfálticos de base foram modificados com o copolímero SBS com ou sem a presença dos ácidos. Os ensaios avaliados foram penetração, ponto de amolecimento, viscosidade rotacional, ensaio de fluência e recuperação sob tensão múltipla (MSCR), LAS modificado, e estabilidade à estocagem. De maneira geral, a adição dos modificadores melhora o comportamento dos ligantes asfálticos, tendo em vista que melhoram a recuperação, ficam menos sensíveis à deformação permanente e são mais tolerantes à fadiga. Pode-se constatar que o tipo de ligante asfáltico de base influencia diretamente o comportamento dos materiais, independente dos modificadores adicionados. A adição dos modificadores foi mais expressiva no ligante asfáltico proveniente da Replan. Em relação aos ácidos, conclui-se que a adição do PA85 foi satisfatória, uma vez que apresentou comportamento semelhante ao do ácido PPA, porém com efeito um pouco menos expressivo. A adição dos ácidos melhorou a estabilidade à es tocagem dos ligantes asfálticos modificados, e pode-se constatar que a adição de um destes modificadores permite a redução do teor do copolímero SBS. Quanto à capacidade de suporte de tráfego, a adição de altos teores desses modificadores não é viável, devido ao fato de apresentarem suporte similar a ligantes asfálticos modificados com teores baixos e intermediários. / The addition of modifiers in asphalt binder to improve its behavior is a common practice nowadays. There are many types of modifiers, among them the most used in the Brazilian federal highways is the SBS copolymer. In addition to polymers, polyphosphoric acid (PPA) is another type of modifier that produces good characteristics when added to asphalt binder. Despite the positive effects, it is known that this material is imported, which makes its purchase as well as the final product very expensive. Considering the opportunity to work with a product with the same basic component, but of national origin, which would make it the most affordable modifier, the present study decided to verify the behavior of the addition of phosphoric acid (PA85) to the asphalt binder. Therefore, the purpose of this thesis is to analyze the effect of the addition of phosphoric acid to the rheological behavior of pure and modified asphalt binders with the SBS copolymer. For both, empirical and rheological tests were conducted in order to verify the behavior of these materials/substances. In the first part of the research, two asphalt binders base CAP50/70 asphalt binder graded PG64-XX (Replan and Lubnor) were modified only with the addition of the two acids (PPA and PA85), while in the second part of these asphalt binders base were modified with SBS copolymer with or without the presence of acids. The tests evaluated penetration, softening point, rotational viscosity, multiple stress creep and recovery (MSCR), LAS modified, and storage stability. In general, the addition of the modifiers enhances the behavior of the asphalt binders, for they improve recovery, are less susceptible to rutting and are more resistant to fatigue. It was found that the type of asphalt binder base directly influences the behavior of materials, regardless of added modifiers. The addition of modifiers was greater in the asphalt binder from the Replan. Compared to acids, it is concluded that the addition of PA85 was satisfactory, since the performance was similar to the PPA acid behavior, but with slightly less significant effect. The addition of acid improved the storage stability of the modified asphalt binders, and it was found that the addition of such modifiers allows reducing the content of the SBS copolymer. Regarding the ability to support traffic, the addition of high levels of these modifiers is not feasible because they show similar support for modified asphalt binders with low and intermediate levels.

Trata-se de uma investigação sobre a influência de alguns fatores no comportamento mecânico de misturas asfálticas densas quanto à estabilidade e à flexibilidade. Foram testados três tipos de ligantes, asfalto convencional, modificado com 4,5% de SBS e modificado com 20% de borracha reciclada de pneu; duas distribuições granulométricas do agregado, centros das faixas B e C do DNER (1997); quatro teores de ligante, escolhidos com base nos valores de volumes de vazios e espessuras de película almejados; três condições de envelhecimento a longo prazo, mistura não envelhecida, envelhecida em estufa ventilada a 85ºC por 5 dias e envelhecida por exposição ao tempo por 4 meses; e duas condições de envelhecimento a curto prazo, mistura não envelhecida e envelhecida em estufa ventilada a 135ºC por 4 horas. A técnica de planejamento e análise de experimentos fatoriais fracionários assimétricos foi utilizada para a consideração simultânea dos fatores citados. O comportamento mecânico das misturas foi avaliado através dos ensaios de resistência à tração, módulo de resiliência e fluência por compressão uniaxial estática e dinâmica. A análise de variância dos resultados permitiu a identificação dos fatores com influência significativa nas respostas dos ensaios. O modo como cada fator interferiu nas propriedades apresentadas pelas misturas foi estabelecido através da construção de modelos estatísticos de comportamento. Os resultados mostraram que a adição de modificadores ao asfalto melhora a resistência das misturas à fadiga e à deformação permanente. Os processos de envelhecimento aumentaram os módulos de resiliência das misturas mas diminuíram sua capacidade de recuperação elástica, o que significa queda de resistência à fadiga. Por outro lado, a resistência a deformações permanentes das misturas, inclusive daquelas compostas por asfaltos modificados, aumentou com o envelhecimento. / This work deals with the influence of some factors in the mechanical behavior of asphalt mixtures stability and flexibility. Three types of binders were tested, conventional one, modified with 4,5% of SBS and modified with 20% of recycled tire rubber. Two aggregate gradations were tested, center of B and C gradations of DNER (1997). Four binder contents were chosen based on the air voids and film thickness. Three types of long-term aging were tested, not aged, aged in a forced-draft oven for 5 days at 85ºC and aged through weather exposition for the period of 4 months. Two types of short-term aging were tested; not aged and aged in a forced-draft oven for 4 hours at 135ºC. The technique of design and analysis of asymmetric fractional factorial experiments was used for the simultaneous analysis of the factors. The mechanical behavior of the mixtures was evaluated based on indirect tensile strength test, resilient modulus test and static and dynamic creep tests. The results of the analysis of variance allowed the identification of factors with significant influence in the answers. The influence of the factors in the mixtures properties was established through statistical models of behavior. The results showed that modified binders improves the mixtures resistance in relation to fatigue and to permanent deformation. The aging processes increased the resilient modulus of the mixtures, but also decreased its capacity of elastic return, resulting in a lost of fatigue resistance. On the other hand, the permanent deformation resistance of the aging mixtures increased, including the mixtures with modified binders cases.

Les structures avec des lieurs sont très communes en informatique. Les langages de programmation et les systèmes logiques sont des exemples de structures avec des lieurs. La manipulation de lieurs est délicate, de sorte que l’écriture de programmes qui ma- nipulent ces structures tirerait profit d’un soutien spécifique pour les lieurs. L’environ- nement de programmation Beluga est un exemple d’un tel système. Nous développons et présentons ici un compilateur pour ce système. Parmi les programmes pour lesquels Beluga est spécialement bien adapté, plusieurs peuvent bénéficier d’un compilateur. Par exemple, les programmes pour valider les types (les "type-checkers"), les compilateurs et les interpréteurs tirent profit du soutien spécifique des lieurs et des types dépendants présents dans le langage. Ils nécessitent tous également une exécution efficace, que l’on propose d’obtenir par le biais d’un compilateur. Le but de ce travail est de présenter un nouveau compilateur pour Beluga, qui emploie une représentation interne polyvalente et permet de partager du code entre plusieurs back-ends. Une contribution notable est la compilation du filtrage de Beluga, qui est particulièrement puissante dans ce langage. / In computer science, structures with variable binders are very common. Program- ming languages and logical frameworks are examples of structures with binders. Thus writing programs that deal with these kinds of data benefits with explicit support for data binding. The Beluga programming environment is an example of such a system. In this work we develop and present a compiler for the system. Many of the programs that Beluga is specially well suited for writing can benefit from a compiler. For example, some of the kinds programs that would benefit more are type-checkers, compilers and interpreters that take advantage of the binder support and dependent types present in the language, and also require a reasonably fast run-time. Our goal in this work, is to present a compiler for the Beluga system, that uses a very versatile internal representation that helps with the development of the system, and allows a sharing of code between several back-ends. Furthermore, we present a way of compiling the uniquely powerful pattern language supported by Beluga.

La maladie cœliaque ou sprue cœliaque est une intolérance au gluten. Il s’agit d’une maladie inflammatoire de l’intestin liée à l’ingestion de gluten chez des personnes génétiquement susceptibles. Ce désordre présente une forte prévalence puisqu’il touche 1 % de la population mondiale. En l’état actuel des choses, il n’existe aucun outil pharmacologique pour traiter ou pallier à cette maladie. Cependant, grâce aux avancées dans la compréhension de sa pathogenèse, de nouvelles cibles thérapeutiques ont été identifiées. À l’heure actuelle, le seul traitement efficace consiste à suspendre la consommation de l’agent pathogène, à savoir le gluten. Le gluten est un ensemble de protéines de stockage des céréales contenu dans le blé, l’orge et le seigle. Le gluten du blé se subdivise en gluténines et gliadines. Ce sont ces dernières qui semblent les plus impliquées dans la maladie cœliaque. Les gliadines et ses protéines apparentées (i.e. sécalines et hordéines, respectivement dans le seigle et l’orge) sont riches en prolines et en glutamines, les rendant résistantes à la dégradation par les enzymes digestives et celles de la bordure en brosse. Les peptides résultant de cette digestion incomplète peuvent induire des réponses immunitaires acquises et innées. L’objectif principal de cette thèse était de tester un nouveau traitement d’appoint de la maladie cœliaque utile lors de voyages ou d’évènements ponctuels. Dans les années 80, une observation italienne montra l’inhibition de certains effets induits par des gliadines digérées sur des cultures cellulaires grâce à la co-incubation en présence de mannane: un polyoside naturel composé de mannoses. Malheureusement, ce traitement n’était pas applicable in vivo à cause de la dégradation par les enzymes du tractus gastro-intestinales du polymère, de par sa nature osidique. Les polymères de synthèse, grâce à la diversité et au contrôle de leurs propriétés physico-chimiques, se révèlent être une alternative attrayante à ce polymère naturel. L’objectif de cette recherche était d’obtenir un polymère liant la gliadine, capable d’interférer dans la genèse de la maladie au niveau du tube digestif, afin d’abolir les effets délétères induits par la protéine. Tout d’abord, des copolymères de type poly (hydroxyéthylméthacrylate)-co-(styrène sulfonate) (P(HEMA-co-SS)) ont été synthétisés par polymérisation radicalaire contrôlée par transfert d’atome (ATRP). Une petite bibliothèque de polymères a été préparée en faisant varier la masse molaire, ainsi que les proportions de chacun des monomères. Ces polymères ont ensuite été testés quant à leur capacité de complexer la gliadine aux pH stomacal et intestinal et les meilleurs candidats ont été retenus pour des essais cellulaires. Les travaux ont permis de montrer que le copolymère P(HEMA-co-SS) (45:55 mol%, 40 kDa) permettait une séquestration sélective de la gliadine et qu’il abolissait les effets induits par la gliadine sur différents types cellulaires. De plus, ce composé interférait avec la digestion de la gliadine, suggérant une diminution de peptides immunogènes impliqués dans la maladie. Ce candidat a été testé in vivo, sur un modèle murin sensible au gluten, quant à son efficacité vis-à-vis de la gliadine pure et d’un mélange contenant du gluten avec d’autres composants alimentaires. Le P(HEMA-co-SS) a permis de diminuer les effets sur les paramètres de perméabilité et d’inflammation, ainsi que de moduler la réponse immunitaire engendrée par l’administration de gliadine et celle du gluten. Des études de toxicité et de biodistribution en administration aigüe et chronique ont été réalisées afin de démontrer que ce dernier était bien toléré et peu absorbé suite à son administration par la voie orale. Enfin des études sur des échantillons de tissus de patients souffrants de maladie cœliaque ont montré un bénéfice therapeutique du polymère. L’ensemble des travaux présentés dans cette thèse a permis de mettre en évidence le potentiel thérapeutique du P(HEMA-co-SS) pour prévenir les désordres reliés à l’ingestion de gluten, indiquant que ce type de polymère pourrait être exploité dans un avenir proche. / Celiac Disease or celiac sprue is identified as a gluten intolerance. It is an inflammatory disorder of the intestine triggered by the ingestion of gluten in genetically susceptible individuals. This condition is highly prevalent because it affects up to 1% of the worldwide population. Nowadays, there is no pharmacological treatment available to treat or off set to the disease. Due to the huge progress in the understanding of the pathogenesis, new therapeutic targets have been discovered. At the present time, the only effective treatment remains the strict lifelong abandonment of the pathogen agent, gluten. Gluten encompasses the storage proteins in wheat, rye and barley. The wheat gluten is divided into glutenins and gliadins. The latter seem to be the most important trigger in the celiac disease. Gliadins and the related proteins (i.e. secalins and hordeins, respectively in rye and barley) are rich in prolin and glutamin residues, conferring them to be resistant by enzymatic digestion. The resulting peptides of the incomplete process may set off the inflammatory reaction. The main objective of this thesis was to test a new supportive therapy in treating celiac disease, useful in punctual occasion (i.e. during travel or social event when the gluten-free property cannot be ascertained). In the 1980’s, inhibition of some gliadin-induced effects on cell cultures were observed owing to mannan co-incubation. However, this compound, due to his osidic nature, may be cleaved by digestive enzymes in vivo. Synthetic polymers prove to be an attracting alternative owing to the tunability of their physical and chemical properties. The goal of this study was to obtain a polymeric gliadin-binder, interferring with the pathogenesis of the celiac disease in the gastro-intestinal tract, to abrogate the gliadin induced effects. Atom transfer radical polymerization was used to synthesize copolymers of the type poly (hydroxyethylméthacrylate)-co-(styren sulfonate) (P(HEMA-co-SS)). A small library of polymers varying in their molecular weight and in their constituting monomers ratio was prepared. The ability of these polymers to sequester gliadin was assessed at stomacal and intestinal pH. The best candidates were further evaluated in cell cultures. Our results revealed that a selective binding was obtained with the P(HEMA-co-SS) (45:55 mol%, 40 kDa). This compound abolished gliadin-induced effects on various cell lines. In addition, gliadin digestion was hindered, suggesting a decrease in the formation of immunogenic peptides known to trigger the diseases. In vivo experiments were additionally carried out on a murine model of gluten-sensitivity with this polymeric candidate. Its efficacy towards pure gliadin and gluten containing food was tested. P(HEMA-co-SS decreased gliadin–induced effect on permeability and inflammatory parameters and modulated the immune response due to gliadin/gluten gavage. Toxicity and biodistribution studies following acute and chronic administration were performed on murine to demonstrate that the polymer was well tolerated and not absorbed after oral administration. Finally, biopsies of patients suffering from CD disease exhibited therapeutic benefit of the polymer. Altogether, the results presented in this thesis evidenced the potential of P(HEMA-co-SS) to prevent the gluten-induced disorder, indicating that this type of polymer may be useful in a near future.