Nesse trabalho obtiveram-se membranas de poli(imida) ramifricada com tamanho de poro ajustável usando melamina como agente de ramificação e indutor da formação de poros. Estas poli(imidas) foram sulfonadas usando-se ácido sulfúrico concentrado eficazmente. Finalmente, obtivemos membranas de carbono molecular por pirólise sob atmosfera inerte das poli(imidas) ramificadas. Estas membranas apresentam canais micrométricos paralelos à superfície e uma estrutura assimétrica constituída de uma camada densa filtrante e uma camada com canais micrométricos paralelos à superfície altamente ordenados. A membrana apresentou boa estabilidade química frente ao ataque por radicais hidroxila gerados via reação de Fenton. / In this work was obtained branched poly(imides) with tuned pore size using melamine as branching and pore induction agent. The poly(imides) with were efficiently sulfonated with concentrated sulfuric acid. Finally, the molecular sieve carbon membranes were obtained by pyrolysis under inert atmosphere of the branched poly(imides). These membranes have an asymmetric structure with a dense filtering layer and a porous layer with highly ordered channels standing parallel to the surface. These membranes display good chemical stability toward hydroxyl radical attack produced by Fenton reaction.

INTRODUÇÃO: O destreinamento físico está relacionado com alterações moleculares associadas à perda de massa muscular, rápido acréscimo da massa adiposa, ganho de peso e resistência à insulina. Estudos apontam que a restrição calórica reduz a gordura corporal, contudo, associada com a inatividade física, altera o metabolismo proteico acelerando o catabolismo muscular. Nesse sentido, estudos com suplementação de aminoácidos essenciais, em especial a leucina, observam aumento na síntese proteica e redução da degradação proteica em situações de restrição ou recuperação nutricional. Dessa forma sugere-se que a restrição calórica associada à suplementação com leucina poderia atenuar os efeitos desencadeados pelo destreinamento físico. OBJETIVO: Investigar a influência da suplementação crônica de leucina na via de sinalização da síntese proteica e degradação proteica no tecido muscular a partir de parâmetros moleculares em ratos destreinados, submetidos à restrição calórica. MÉTODOS: Foram utilizados 64 ratos Sprague-Dawley machos e adultos, inicialmente distribuídos em 2 grupos: Controle (CON) (n = 16) representados pelos animais sedentários, e Treinamento (TREIN) (n = 48) que foram submetidos ao treinamento em esteira ergométrica durante oito semanas. Após esse período, os animais foram redistribuído em 6 grupos: Sedentário (SED), Treinamento (TREIN), Destreinamento (DT), Destreinamento + Leucina (LEU), Destreinamento + Restrição Calórica (DTRC) e Destreinamento + Restrição Calórica + Leucina (DTRC + LEU). Foram analisados massa corporal, consumo da ração, composição corporal, sensibilidade a insulina bem como os marcadores de inflamação (IL-6; IL-10; MCP-1; TNF-α; 1L-1α; PAI-1; leptina; adponectina) e parâmetros moleculares como genes e proteínas envolvidas na via de sinalização da síntese protéica (mTOR, P-4EBP1, P-s6K1 e eIF4E); degradação proteica (MAFBx e MURF) além de transportadores de aminoácidos (LAT-1 e SNAT 2 e CD98). ESTATÍSTICA: Os valores foram expressos em média e desvio padrão. As comparações entre os grupos após o período de destreinamento físico foram avaliadas por meio de análise de variância (ANOVA), seguida do teste de Tukey. Em todas as análises foi considerado nível de significância de 5%. A análise estatística foi realizada no software SPSS versão 17.0. RESULTADOS: Em relação à composição corporal, foi observada diferença estatisticamente significativa na gordura corporal e massa livre de gordura entre os grupos DTRC e DTRC+LEU, em relação aos demais grupos experimentais. Porém não houve diferença estatística entre o DTRC e DTRC+LEU. No entanto não foi observada diferença estatisticamente significativa quando avaliado a proteína da carcaça. Em relação aos parâmetros moleculares, não foi observada diferença estatisticamente significativa entre os grupos, quando avaliada a expressão de proteínas relacionadas com a via de sinalização de síntese proteica (mTOR, P-4EBP1, P-s6K1 e eIF4E) e transportadores de leucina (LAT- 1;SNAT-2;CD(98). Quanto avaliada a expressão gênica da via de degradação, foi observada uma menor expressão do gene MURF quando suplementado com leucina, porém sem diferença estatisticamente significativa. CONCLUSÃO: A restrição calórica associada com a suplementação com leucina foi efetiva na redução da gordura corporal, e aumento da massa livre de gordura, porém não houve diferença estatisticamente significante entre os dois grupos DTRC e DTRC+LEU, tampouco quando avaliada a proteína da carcaça desses animais. Dessa forma, pode-se concluir que a suplementação crônica com leucina não reverteu os efeitos desencadeados pelo destreinamento físico, e, além disso, não foi suficiente para alterar os parâmetros moleculares envolvidos na via de sinalização de síntese e degradação proteica desses animais. / INTRODUCTION: Physical detraining is related to molecular changes associated with loss of muscle mass, rapid increase in fat mass, weight gain and insulin resistance. Studies show that caloric restriction reduces body fat; however, associated with physical inactivity, it alters protein metabolism accelerating muscle catabolism. Accordingly, studies with supplementation of essential amino acids, particularly leucine, observed increase in protein synthesis and reduced protein degradation in situation of nutritional restriction or recovery. Thus, it is suggested that caloric restriction associated with leucine supplementation could attenuate the effects triggered by physical detraining. OBJECTIVE: To investigate the influence of chronic leucine supplementation in the signaling pathway of protein synthesis and degradation in muscle tissue from molecular parameters in detrained rats, subjected to caloric restriction. METHODS: Sixty-four adult male and female Sprague-Dawley rats were used, initially divided into 2 groups: Control (CON) (n = 16) represented by sedentary animals, and Trained (TRAIN) (n = 48) who underwent treadmill training for eight weeks. After this period, the animals were re-distributed into 6 groups: Sedentary (SED), Trained (TRAIN), Detrained (DT), Detrained + Leucine (LEU), Detrained + Caloric Restriction (DTRC) and Detrained + Caloric Restriction + Leucine (DTRC + LEU). Body mass, food consumption, body composition, insulin sensitivity were analyzed, as well as inflammation markers (IL-6; IL-10; MCP-1; TNF-α; 1L-1α; PAI-1; leptin; adiponectin) and molecular parameters, such as genes and proteins involved in signaling pathways of protein synthesis (mTOR, P-4EBP1, P-s6K1 and eIF4E); protein degradation (MAFBx and MURF) and also amino acid transporters (LAT-1, SNAT 2 and CD98). STATISTICAL ANALYSIS: Values were expressed as mean and standard deviation. Analysis of variance (ANOVA) was used for comparisons between groups after physical detraining, followed by Tukey\'s test. A 5% significance level was considered in all analyses. Statistical analysis was performed using SPSS software, version 17.0. RESULTS: In relation to body composition, a statistically significant difference was observed in body fat and fat free mass between groups DTRC and DTRC+LEU, compared with other experimental groups. However, there was no statistical difference between groups DTRC and DTRC+LEU. Nevertheless, no statistically significant difference was found when carcass protein was assessed. In relation to molecular parameters, no statistically significant difference was observed between groups, when protein expression related to the signaling pathway of protein synthesis (mTOR, P-4EBP1, P-s6K1 and eIF4E) and leucine transporters (LAT-1;SNAT-2;CD(98) was assessed. When gene expression of the degradation pathway was investigated, a lower expression of gene MURF was found with leucine supplementation; however, this was not statistically different. CONCLUSION: Caloric restriction associated with leucine supplementation was effective in reducing body fat, and increasing fat free mass; however, no statistically significant difference was found between groups DTRC and DTRC+LEU, nor when carcass protein of these animals was assessed. Therefore, it was concluded that chronic leucine supplementation did not reverse the effects triggered by physical detraining and, in addition, it was not sufficient to change the molecular parameters involved in the signaling pathway of protein synthesis and degradation of these animals.

Le polypropylène isotactique est un thermoplastique semi-cristallin de grande consommation présentant de nombreux avantages tels que sa résistance chimique, sa haute température de fusion ou sa rigidité. Néanmoins, sa faible résistance à l'état fondu le rend inadapté à certains procédés de mis en forme. La présence branchements longs au sein de la structure polymérique permet toutefois de renforcer significativement les propriétés d'un polymère dans son état fondu. De ce fait, de nombreuses approches ont été développées afin de générer du polypropylène branché (irradiation, addition de peroxides, greffages etc.). La catalyse de polymérisation par les métallocènes est d'une grande flexibilité dès qu'il s'agit de concevoir des polymères de spécialité. De plus, la rhéologie moléculaire est un outil indispensable à la compréhension des liens entre les propriétés viscoélastiques et la structure macroscopique des polymères (distribution des masses, topologie des branchements, etc.). Grâce à l'alliance de ces deux expertises, les travaux de cette thèse se sont focalisés sur la caractérisation structurelle, thermique et rhéologique de matériaux de type LCB-iPP obtenus par catalyse des zirconocènes {Cp'CR2Flu}ZrCl2. L'objectif premier était de jouer sur la génération des branchements longs en modifiant les conditions opératoires de polymérisation du propylène. En parallèle, un gros travail analytique a été fourni afin de développer des protocoles rhéologiques fiables et adaptés. Suite à ces résultats, nous avons démontré qu'un mécanisme via génération/incorporation de macro-α-oléfines était à l'origine de la génération de LCB-iPP, ce qui a permis la synthèse de polypropylènes hautement branchés et de vérifier l'effet positif des branchements longs sur le comportement viscoélastique des matériaux finaux. / Because of its high melting point, high tensile strength, stiffness and chemical resistance, isotactic polypropylene has one of the leading and fast growing thermoplastic polymers in the world. However, commercial PPs usually have relatively low melt strength, which limits their use in applications such as blow molding. Since long-chain branching (LCB) is known to enhance the melt properties of a polymer, several approaches have been developed to make branched polypropylenes (electron beam irradiation, peroxide curing, grafting etc.). Development of metallocene technology provides unprecedented flexibility in polymer design. Many structural features, including LCB, can now be introduced into polymers. In this work, long chain branched isotactic polypropylene (LCB-iPP) was synthesized using {Cp'CR2Flu}ZrCl2 metallocene catalyst and fully characterized (rheological, spectroscopic and thermal analysis). The branching (macro-α-olefins having predominantly vinyl-terminated chain end) was in situ generated and further incorporated by the same catalyst system to obtain LCB-PP/Linear PP blends. The LCB content was increased using a novel tandem catalysts system for converting propylene alone to isotactic polypropylene with long branches which exhibit enhanced melt properties.

<p>A series of novel methods for detection of known sequence variants in DNA, in particular single nucleotide polymorphism, using padlock probes and rolling circle replication are presented. DNA probes that can be circularized – padlock probes – are ideal for rolling circle replication. Circularized, but not unreacted probes, can generate powerful signal amplification by allowing the reacted probes to template a rolling circle replication (RCR) reaction. However, when hybridized and ligated to a target DNA molecule with no nearby ends, the probes are bound to the target sequence, inhibiting the RCR reaction is. This problem can be solved by generating a branched DNA probe with two 3’ arms such that the probes may be circularized while leaving the second 3’ arm as a primer for the RCR reaction. We describe how T4 DNA ligase can be used for efficient construction of DNA molecules having one 5’ end but two distinct 3’ ends that extend from the 2’ and 3’ carbons of an internal nucleotide. An even stronger approach to circumvent the topological problem that can inhibit RCR is to restriction digest the template downstream of the padlock recognition site. By using Phi 29 DNA polymerase with efficient 3’ exonuclease and strand displacement activity, the template strand can then be used to prime the RCR reaction. The amplified molecule is contiguous with the target DNA, generating an anchored localized signal. The kinetics of the reaction was investigated by following the reaction in real-time using molecular beacon probes. Localized RCR signal were obtained on DNA arrays, allowing detection of as little as 104-105 spotted molecules, of either single- or double-stranded M13 DNA, in a model experiment. We have also established a serial rolling circle amplification procedure. By converting rolling circle products to a second and even third generation of padlock probes the signal was amplified thousand-fold per generation. This procedure provides sufficient sensitivity for detection of single-copy gene sequences in 50 ng of human genomic DNA, and large numbers of probes were amplified in parallel with excellent quantitative resolution.</p>

A series of novel methods for detection of known sequence variants in DNA, in particular single nucleotide polymorphism, using padlock probes and rolling circle replication are presented. DNA probes that can be circularized – padlock probes – are ideal for rolling circle replication. Circularized, but not unreacted probes, can generate powerful signal amplification by allowing the reacted probes to template a rolling circle replication (RCR) reaction. However, when hybridized and ligated to a target DNA molecule with no nearby ends, the probes are bound to the target sequence, inhibiting the RCR reaction is. This problem can be solved by generating a branched DNA probe with two 3’ arms such that the probes may be circularized while leaving the second 3’ arm as a primer for the RCR reaction. We describe how T4 DNA ligase can be used for efficient construction of DNA molecules having one 5’ end but two distinct 3’ ends that extend from the 2’ and 3’ carbons of an internal nucleotide. An even stronger approach to circumvent the topological problem that can inhibit RCR is to restriction digest the template downstream of the padlock recognition site. By using Phi 29 DNA polymerase with efficient 3’ exonuclease and strand displacement activity, the template strand can then be used to prime the RCR reaction. The amplified molecule is contiguous with the target DNA, generating an anchored localized signal. The kinetics of the reaction was investigated by following the reaction in real-time using molecular beacon probes. Localized RCR signal were obtained on DNA arrays, allowing detection of as little as 104-105 spotted molecules, of either single- or double-stranded M13 DNA, in a model experiment. We have also established a serial rolling circle amplification procedure. By converting rolling circle products to a second and even third generation of padlock probes the signal was amplified thousand-fold per generation. This procedure provides sufficient sensitivity for detection of single-copy gene sequences in 50 ng of human genomic DNA, and large numbers of probes were amplified in parallel with excellent quantitative resolution.

The dibenzo[a,c]phenazine dimers with the branched alkoxy chain of disc were synthesized simplely. We now show on the synthesis of this series of compounds which were found to exhibit mesomorphism as determined by polarized optical microscopy (POM), differential scanning calorimetry (DSC) and X-ray diffraction (XRD). These materials showed polymeso- morphism and glass formation. In our investigations, we also observed that the effects of branched chains on dibenzo[a,c]phenazine enhanced columnar liquid crystal phase stability indeed.

Utilization of methylaluminoxane (MAO) activated metallocene and constrained geometry (CGC) olefin polymerization catalysts containing fluorenyl or octamethyloctahydrodibenzofluorenyl (Oct) moieties has yielded three series of syndiotactic copolymers of propylene with higher a-olefins. The melting temperatures of these polymers were analyzed, and found to correspond directly with the mole percent incorporation of comonomer, as well as with the frequency of stereoerrors in the polymers. Further analysis indicated that rmrr stereoerrors, a result of site epimerization, occur in close proximity to the incorporated comonomers. The MAO-activated fluorenyl/Oct-containing metallocene and CGC catalysts were further utilized to produce syndiotactic samples of poly(1-butene) (s-PB) and poly(1- pentene) (s-PPe). The syndiotacticity of the samples was quantified by 13C NMR and the melting temperatures determined by DSC. The samples of s-PB and s-PPe produced by Me2Si( h1-C29H36)( h1-N-tBu)ZrCl2·OEt2 (Oct-CGC) were found to melt at higher temperatures (55.9 and 43.1 °C, respectively) than any previously reported samples. The MAO-activated Oct-CGC was also used to produce polyethylene samples at a variety of polymerization temperatures and pressures. All of the samples were found to contain an unprecedented degree of branching (13-65 total branches per 1000 carbon atoms) for an early transition metal single-site catalyst. The branches were found to be almost exclusively of two or greater than five carbon atoms in length, and the levels of the longer branches could be controlled by varying the polymerization conditions. The number of ethyl branches was roughly 5 per 1000 carbon atoms for all samples. Finally, a binary catalyst system comprising the Oct-CGC and a chromium-based ethylene trimerization catalyst, ((tBuSCH2CH2)2NH)CrCl3, was developed. This MAOactivated catalyst system could be tuned to produce polyethylene samples with 17-49 total branches per 1000 carbon atoms. Between 4 and 16 of these branches were found to arise from incorporation of 1-hexene produced by the chromium oligomerization catalyst. Adjusting the ratios of oligomerization catalyst, polymerization catalyst, and activator was found to allow rational control over the branch content of the polymers. The branching levels could also be varied by altering the time between injection of the oligomerization and polymerization catalysts into the system.

published_or_final_version / Zoology / Master / Master of Philosophy

Obesity is an ever-growing concern in the developed world that carries with it a plethora of health issues. For example, obesity increases an individual’s risk for Type 2 Diabetes and cardiovascular disease. Pregnancy is a vital time for a woman to maintain optimal health, both for her own benefit as well as that of her offspring, and yet almost half of women in the United States who are of age to bear children are overweight or obese. In mice, we found that offspring born to dams fed a high fat diet did not have impaired glucose tolerance, contrary to our hypothesis. In addition, we challenged the offspring with a high fat diet, and found no difference in glucose tolerance as a result of maternal diet. Exercise is at the opposite end of the wellness spectrum – individuals who exercise experience many health benefits. Even overweight or obese individuals who exercise without losing weight have improved insulin sensitivity, for example. Studies have previously used voluntary running and found that offspring born to exercised dams have improved glucose tolerance. With the goal of controlling variable running times and distances, we developed a novel model of controlled exercise and have shown that it is a safe intervention that warrants further study. In addition, many individuals choose to take dietary supplements for various reasons. Branched chain amino acids (BCAAs) are a common dietary supplement that have been shown to increase lean mass, and may be implicated in glucose metabolism. We supplemented female mice with BCAAs for 16 weeks and found that exercise plus BCAAs improved body composition compared to sedentary control-diet fed animals, when exercise alone did not. In summary, we herein explore a number of health behaviors in female mice, both negative treatments such as consumption of a high fat diet and positive interventions such as exercise and BCAA supplementation, and the impact that they may have on the female animal and/or her offspring.

We will discuss what is known about the construction of contact structures via branched covers, emphasizing the search for universal transverse knots. Recall that a topological knot is called universal if all 3-manifold can be obtained as a cover of the 3-sphere branched over that knot. Analogously one can ask if there is a transverse knot in the standard contact structure on S³ from which all contact 3-manifold can be obtained as a branched cover over this transverse knot. It is not known if such a transverse knot exists.