Thesis (Ph. D.)--University of Missouri-Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves 327-343). Also available on the Internet.

Thesis (Ph.D.)--Victoria University of Wellington, 2008. / Embargoed until 16 March 2011. Includes bibliographical references.

Thesis (Ph. D.)--University of Chicago, Dept. of Chemistry, March 1999. / Includes bibliographical references. Also available on the Internet.

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Orientador: Gisele Zoccal Mingoti / Banca: Fernanda da Cruz Landim / Banca: Joaquim Mansano Garcia / Resumo: O objetivo desse estudo foi avaliar os efeitos da suplementação do meio com diferentes fontes de macromoléculas, com bloqueadores da meiose e com antioxidantes durante o transporte de oócitos bovinos por 6 horas sobre: 1) progressão da maturação nuclear; 2) maturação citoplasmática e 3) competência no desenvolvimento e criotolerância dos embriões produzidos. Para tanto, o meio de transporte de oócitos foi suplementado com bloqueadores da meiose (forscolina e IBMX; Experimento 1) ou com diferentes tipos de macromoléculas (SFB ou BSA; Experimento 2), sendo que estes tratamentos ainda receberam ou não a suplementação com antioxidantes (mistura de cisteína, cisteamina e catalase). Os oócitos foram incubados em incubadora portátil (Minitub®) para simulação de transporte. Posteriormente, foram submetidos à maturação in vitro (MIV) em incubadora a 5% de CO2 em ar até completar 24h e, em seguida, foram fecundados e os prováveis zigotos foram cultivados in vitro durante 7 dias. Foi feito um grupo controle adicional no experimento I: MIV em incubadora com 10% de SFB por 24h. No experimento II foram feitos dois grupos controle adicionais MIV em incubadora com 10% de SFB por 24h sem e com antioxidantes (cisteína, cisteamina e catalase). Nos experimentos 1 e 2 foi avaliada a cinética da maturação nuclear e a maturação citoplasmática (através do posicionamento de mitocôndrias, do potencial de membrana mitocondrial e do conteúdo intracelular de espécies reativas do oxigênio) após o transpor... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this study was to evaluate the effects of supplementation of the medium with different sources of macromolecules with blockers of meiosis and antioxidants during transport of bovine oocytes for 6 hours on: 1) progression of nuclear maturation; 2) cytoplasmic maturation and 3) competence in the development and cryotolerance of embryos produced. Therefore, the medium of transport oocytes was supplemented with blocking of meiosis (forskolin and IBMX; Experiment 1) or with different types of macromolecules (FCS or BSA; Experiment 2), and these treatments yet received or not antioxidant supplementation (mixture of cysteine, cysteamine and catalase). Oocytes were incubated in a portable incubator (Minitub®) for transport simulation. Posteriorly were submitted in vitro maturation (IVM) in incubator at 5% CO2 in air until to complete 24 hours and then were fertilized and presumptive zygotes were cultured in vitro for 7 days. Has been made an additional control group in the experiment I: MIV incubator with 10% FCS for 24 hours (Control). In the second experiment were performed two additional control groups: IVM incubator with 10% FCS for 24 hours (Control); and IVM in an incubator with 10% FCS and antioxidants (cysteine, cysteamine and catalase) for 24 hours (Contr+Atx). In Experiments 1 and 2 were evaluated after nuclear maturation kinetics and cytoplasmic maturation (made by positioning mitochondria, the mitochondrial membrane potential and intracellular content of ... (Complete abstract click electronic access below) / Mestre

No description available.

The aim of this thesis is to investigate the structural and thermodynamic properties of biologically and technological relevant macromolecules when placed at soft interfaces. In particular two amphiphilic macromolecules characterized by different topologies have been investigated namely amphiphilic dendrimers and linear block copolymers. This goal is achieved using a multiscale approach which includes all-atom, united atom and coarse grained models by means of molecular dynamic simulations.Amphiphilic dendrimers have shown to be promising building blocks for a range of interfacial materials and can be used in applications such as surface-base sensors or surface nanopatterning. In this part of the thesis by means of all-atom molecular dynamics simulations, we investigated the structure and stability of alkyl-modified polyamido-amide (PAMAM) dendrimers at the air/water interface as a function of the number and the relative position of the modified end groups. We found that the PAMAM dendrimer with all terminal groups functionalized is more stable at the interface than the Janus dendrimer, where only half the amine groups are modified. These results indicate that monolayers of fully functionalized molecules could be as stable as (or more stable than) those self-assembled from Janus molecules.The second part of the thesis is devoted to model a particular family of amphiphilic triblock copolymer sold as Pluronics, consisting of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) arranged as PEO–PPO–PEO. There is evidence that this class of amphiphilic materials can be used for different biological applications. A fuller understanding of the molecular mechanisms underpinning their interactions with living cells is essential for ensuring the polymers safety and efficacy in biomedical applications. Using united-atom molecular dynamics simulations and membrane lysis assays, we investigated the relationship between the molecular conformations of a subset of the Pluronic copolymers (L31, L61, L62 and L64) and their haemolytic activity. Our computational studies suggest that the hydrophilic blocks in these copolymers interact with the polar head groups of lipid molecules, resulting in a predicted modification of the structure of the membranes. Parallel membrane lysis assays in human erythrocytes indicate differences in the rates of haemolysis, as a result of incubation with these polymers, which correlate well with the predicted interactions from the atomistic simulations. The computational data thus provide a putative mechanism to rationalize the available experimental data on membrane lysis by these copolymers. The data quantitatively agree with haemoglobin release endpoints measured when copolymers with the same molecular weight and structure as of those modelled are incubated with erythrocytes. The data further suggest some new structure– function relationships at the nanoscale that are likely to be of importance in determining the biological activity of these otherwise inert copolymers.In order to visualise the effect of Pluronics at a length and time scale closer to the experimental one, in the third part of the thesis we developed a coarse-grained model for the amphiphilic copolymers within the framework of the MARTINI forcefield (Marrink et al., J. Phys. Chem. B, 2007, 111, 7812). The MARTINI force field is usually parameterized targeting thermodynamic properties. In addition to this, we further parameterized it based on atomistic simulations validating the parameters against structural properties of the copolymers. The ability of the model to predict several structural and thermodynamic properties of the atomistic system have been explored. The aim of this work is to be able to simulate the polymer/lipid interface at polymer concentration similar to the experimental one.

Nanoparticulas magnéticas de óxido de ferro tem sido amplamente utilizadas em diversas áreas da biotecnologia e biomedicina, tais como no tratamento de câncer, marcação de célula e como agentes de contraste em imagem por ressonância magnética. O intuito deste trabalho foi sintetizar as nanopartículas magnéticas com magnetização de saturação intensificadas via processo do poliol modificado, e usando agentes de superfície para melhorar as propriedades de superfície. Carboximetildextrana, metilpolietilenoglicol (MPEG), quitosana, sílica e 3-aminopropiltrimetoxisilano (APTMS) foram utilizados para a modificação da superfície. Através da microscopia eletrônica de transmissão (TEM), foi obtido que as nanopartículas magnéticas de magnetita obtiveram um diâmetro médio de 5nm, em uma estreita distribuição de tamanho. A difração de raios-X (DRX) indicou a formação de magnetita em todos os sistemas em que o método do poliol modificado foi utilizado. As medidas de espectroscopia no infravermelho (FTIR) evidenciaram a presença de modos de vibração relacionados às macromoléculas e compostos inorgânicos utilizados na modificação de superfície das nanopartículas magnéticas. A TEM das diferentes modificações de superfície mostram a formação de aglomerados dependendo do modificador utilizado. As nanopartículas recobertas com APTMS foram funcionalizadas com ácido fólico, mostrando resultados satisfatórios, porém serão necessárias outras técnicas de caracterização. Para a funcionalização foi determinada a quantidade de amina livre na superfície da nanopartícula recoberta com APTMS e a técnica de UV-Vis determinou um bom resultado. A magnetometria de amostra vibrante (VSM) mostrou comportamentos semelhantes para todas as amostras recobertas em comparação a amostra sem recobrimento. Estes resultados evidenciam que a modificação de superfície foi realizada satisfatoriamente. Os métodos utilizados para realizar a mudança para hidrofóbica a superfície inicialmente hidrofílica se mostraram efetivos, porém a quantidade de agentes modificadores deve ser melhor estudada. Portanto, as nanopartículas magnéticas funcionalizadas com diferentes superfícies foram obtidas e possuem um alto potencial para serem utilizadas em aplicações em biomedicina. / Superparamagnetic iron oxides nanoparticles (SPION) have been highlighted in several areas of biotechnology and biomedicine, for example in cancer treatment, in labeling of cells and as contrast agent in magnetic resonance imaging (MRI). The purpose of this study was synthesizing SPION with intensified saturation magnetization by modified polyol process, and using surface agents to enhance the surface properties. Carboxymethildextran, metylpolietileneglycol, chitosan, silica and 3-aminopropyltrimethoxysilane (APTMS) were utilized as surface modifiers. By transmission electron microscopy (TEM), SPION showed narrow particle size distribution, with an average diameter around 5 nm. The X-ray diffraction studies indicated the formation of magnetite in all synthesized systems in which the modified polyol process was utilized. FTIR measurements showed the presence of vibration modes related to the macromolecules and inorganic compounds used to SPION surface modifications. TEM of the different surface modifications showed the agglomerate formation, which depends on the used surface modifier. SPION coated with APTMS was functionalized with folic acid, showing satisfactory results. However other characterization techniques will be necessary for study this modification. Quantity of free amine groups was determinate in the amount coated with APTMS for functionalization, and UV-Vis spectroscopy determinates a good result. Vibrating sample magnetometry (VSM) indicates similar behaviors in all cases against SPION without surface modifiers. These results suggest that the surface modifications were performed satisfactorily. Utilized methods for changing the hydrophobic to hydrophilic surface showed effectives, however, the quantity of surface modifiers should be better studied. Therefore, SPION functionalized with different hydrophilic surfaces were obtained, which possess high potential to be used as devices in biomedical applications.

Yes / Interpolymer complex formation of poly(acrylic acid) with other macromolecules can occur via several mechanisms that vary depending on the pH. At low pH the protonated acid functional group can form bonds with both donor and acceptor moieties, resulting in desolvated structures consisting of two polymers. Complexes were formed in dilute solutions of PAA, functionalised with acenaphthylene, with a range of other polymers including: poly(NIPAM); poly(ethylene oxide) (PEO); poly(dimethylacrylamide) (PDMA); poly(diethyl acrylamide) (PDEAM) poly(vinyl alcohol) (PVA) and poly(vinyl pyrolidinone) (PVP). Fluorescence anisotropy was used to demonstrate complex formation in each case by monitoring the reductions in segmental motion of the chain as the complexes formed. Considerations of the molecular structures of the complexing moieties suggest that solvation energies and pKas play an important role in complex formation.