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Albumin Adsorption: Inferences of Protein Interactions Measured by Sedimentation both Between Species and Induced by DenaturingMcKeon, Kristin Dianne 20 May 2008 (has links)
Biological development and progression are managed by a diverse macromolecular group called proteins. Protein structure results from a complex folding process that leads to a final active form. This protein state is susceptible to changes in the surrounding environment and an incorrect structure can be produced. Changes in the protein conformation can lead to the formation of protein aggregates. Adsorption of proteins onto surfaces is utilized in many research analyses, but is capable of irreversibly changing the protein structure and causing aggregation. Albumin is a plasma protein that adsorbs on many different surfaces because the structure easily rearranges. The structure of albumin once adsorbed has been shown to deteriorate; however, outcomes of both stabilization and aggregation have been found.
A dynamic laser light scattering instrument will be utilized to measure the differences in size and determine the amount of aggregation. Our lab has developed a z-axis translating laser light scattering device (ZATLLS) that has been used to measure the sedimentation velocity of several different materials in solution. In this case, bovine serum albumin (BSA) will be adsorbed onto polystyrene particles and the particle settling velocity determined. The settling solution viscosity and density will also be ascertained, so Stoke's law can infer the average aggregate size of each experiment. BSA-coated polystyrene particles displayed a more controlled settling behavior compared to non-coated polystyrene particles. Although the BSA-coated particles had a smaller sedimentation velocity, a larger aggregate size was found due to the greater solution viscosity. Therefore, the ZATLLS instrument can be employed to measure sedimentation velocities of multiple interactions and the aggregation level inferred.
Although most albumin molecules are remarkably similar, there are subtle differences in amino acid residues, length, and charge. Sedimentation velocities for human serum albumin (HSA) coated polystyrene particles and BSA-coated polystyrene particles only had a small difference. However an almost 50% higher solution viscosity was measured in BSA experiment solutions, and resulted in the slower settling of the larger aggregates compared to HSA-coated particles. Viscosity calibration curves for each albumin species were used to determine the amount of protein desorbed from the particles during the settling process. The larger solution viscosity for BSA-coated particle experiments led to a much larger degree of desorption. HSA was shown to be the more stable albumin species when adsorbed onto polystyrene particles.
Temperature denaturing was performed to aid in the determination of the stability of BSA. Reversible and irreversible conformational changes in BSA were produced at 46ºC and 76ºC respectively. The solutions were cooled to room temperature before adsorption ontopolystyrene particles and the sedimentation velocities measured. A 50% difference in average viscosity between the reversibly and irreversibly changed BSA was found. This caused the larger aggregates formed in the 76ºC BSA experiments to have an almost equivalent sedimentation velocity to those in the reversibly denatured BSA experiments. Average aggregate size for reversibly denatured BSA was well within the ranges found for non-denatured BSA. In conclusion, irreversibly denatured BSA formed larger aggregates and was more likely to desorb from the polystyrene particles than reversibly changed BSA. / Master of Science
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Estruturas tridimensionais fabricadas a partir de esferas quitosana/hidroxiapatita para regeneração óssea.DANTAS, Maria Jucélia Lima. 15 June 2018 (has links)
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Previous issue date: 2016-02-29 / Capes / Estruturas tridimensionais são ferramentas bastante atrativas para a
engenharia de tecidos, mimetizado fisicamente a matrix extracelular natural que
atuam como suportes para o desenvolvimento celular. O objetivo do trabalho foi
produzir estruturas tridimensionais biodegradáveis de quitosana-hidroxiapatitagelatina (CS/HA/G) com diferentes quantidades de HA e avaliar suas propriedades e comportamento in vitro. As estruturas tridimensionais foram produzidas em duas etapas. Numa primeira etapa foram obtidas esferas de CS contendo diferentes quantidades de HA de baixa cristalinidade (20, 50 e 70 % m/m). A HA de baixa cristalinidade foi gerada in situ no interior das esferas de CS. Para isso foi precipitado CaHPO4 em uma dissolução de CS e a suspensão resultante foi conformada na forma de esferas mediante gotejamento em dissolução de Na5P3O10 (TPP) com pH 8-9. As esferas precipitadas foram mantidas sob agitação nesta solução para conseguir a reticulação da CS e a transformação do CaHPO4 em HA, e finalmente
liofilizadas. Para a obtenção das estruturas tridimensionais, as esferas de CS HA foram aglutinadas mediante impregnação com dissolução aquecida (40°C) de 5 % de G, arrefecimento até -18°C e liofilização. As esferas foram caracterizadas mediante microscopia ótica (MO), microscopia eletrônica de varredura (MEV), difração de raios X (DRX), análises termogravimétricas (TG) e espectroscopia de infravermelho (IV). E as estruturas tridimensionais por Porosidade, Grau de Intumescimento (GI), propriedade mecânica de resistência à comrpressão via seco e úmido. O diâmetro médio das esferas segundo os resultados de MO foi de 2,6 ± 0,22mm e 2,8± 0,28mm para CS-HA 20% e CS-HA50%,respectivamente. Já as partículas de CS-HA70%
apresentaram diâmetro médio de 3,9± 0,37mm tendo sua morfologia e
porosidade superficial variou com o conteúdo de HA. A presença de HA de
baixa cristalinidade no interior das esferas foi confirmada pelos resultados DRX
e IV. Nas imagens de MEV foi possível observar que os cristais de HA estão
homogeneamente dispersos no interior das esferas. Os resultados de TG
revelaram boa concordância entre as quantidades de HA projetadas e as
realmente obtidas nas esferas. Os resultados das estruturas tridimensionais
indicam influência pelas diferentes concentrações de hidroxiapatita. Com o
aumento na fração cerâmica observa-se a densificação da superfície, uma
pequena diminuição da porosidade e no grau de intumescimento, como
também um aumento no modulo de elasticidade via seca e uma pequena
diminuição nas propriedades mecânicas via úmido, causado pelo intumescimento da estrutura. O compósito que apresentou resultados mais satisfatórios foi CS-HA20%, exibindo um melhor perfil à porosidade, grau de intumescimento e prorpriedades mecânicas. O estudo sugere que as estruturas tridimensionais necessitam de reajustes para serem aplicadas em regeneração óssea, entretanto podem ser indicadas para aplicações em engenharia de tecidos, em situações preferencialmente ex-vivo, como suportes temporários de células. / Three-dimensional structures are very attractive tools for tissue engineering,
physically mimicked the natural extracellular matrix, these structures act as supports for cell development. The goal was to produce biodegradable threedimensional structures of chitosan-hydroxyapatite-gelatin (CS / HA / L) with different HA contents and evaluate their properties and behavior in vitro. The three-dimensional structures were produced in two steps. In a first step CS were obtained spheres containing different amounts of HA low crystallinity (20, 50 and 70% w / w). The poorly crystalline HA was generated in situ within the CS spheres. To this precipitate was dissolved in CS CaHPO4 and the resulting suspension was shaped in the form of spheres by dripping in dissolution Na5P3O10 (TPP) at pH 8-9. The precipitated spheres were kept in this solution under stirring to obtain the halftone processing and the CS HA CaHPO4
, and finally lyophilized. To obtain the three-dimensional structures, CS-HA spheres
were bonded by impregnation with warm solution (40 ° C) 5% C, cooling to -18 °C and lyophilization. The spheres were characterized by optical microscopy (OM), scanning electron microscopy (SEM), X-ray diffraction (XRD), thermogravimetric analysis (TG) and infrared spectroscopy (IR). And the three dimensional structures by porosity, Degree of Swelling (GI), mechanical property of resistance comrpressão via dry and wet. The average diameter of the spheres according to the results of MO was 2.6 ± 0.22 mm and 2.8 ± 0,28mm CS-20% CSHA50% respectively. Already the particles of CS-HA70% had a mean diameter of 3.9 ± 0,37mm having their morphology and surface porosity varied with the HA content. The presence of low crystallinity with in the
HA spheres was confirmed by the XRD results and IV. In the SEM images was
observed that HA crystals are homogeneously dispersed within the sphere. The
TG data showed good agreement between the HA and the amounts actually
projected on the spheres obtained. The results indicate influences of threedimensional structures for various concentrations of hydroxyapatite. With the
increase in the ceramic fraction is observed densification of the surface, a slight
reduction of the porosity and the degree of swelling, as well as an increase in the modulus of elasticity via a small decrease in the mechanical properties wet route, caused by swelling of the structure. The composite that presented better results was CS-HA20%, showing a better profile to the porosity, degree of swelling and mechanical prorpriedades. The study suggests that threedimensional structures require adjustments to be applied in bone regeneration, but can be suitable for applications in tissue engineering, in situations preferably ex vivo, as temporary supports cells.
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From Particle Condensation to Polymer Aggregation: Phase Transitions and Structural Phases in Mesoscopic Systems: From Particle Condensation to Polymer Aggregation:Phase Transitions and Structural Phases in Mesoscopic SystemsZierenberg, Johannes 17 December 2015 (has links)
Die vorliegende Arbeit befasst sich mit den Gleichgewichtseigenschaften und Phasenübergängen in verdünnten Teilchen- und Polymersystemen, mit einem Fokus auf Teilchenkondensation und Polymeraggregation. Dazu werden sowohl analytische Argumente als auch hochentwickelte Monte Carlo Simulationen verwendet. Um die in dieser Arbeit erreichten Systemgrößen zu simulieren, wurde eine parallele Version der multikanonischen Methode entwickelt. Die Leistungsfähigkeit dieser Erweiterung wird an mehreren relevanten Beispielen demonstriert.
Um Teilchenkondensation und Polymeraggregation in finiten Systemen und in geometrisch beschränkten Strukturen besser zu verstehen, wird der Einfluss von verschiedenen Parametern auf die jeweiligen Übergange untersucht. Dies beinhaltet unter anderem die Systemgröße und Dichte, sowie im Speziellen für semiflexible Polymere deren Steifigkeit. Betrachtet werden sowohl kanonische Observablen (Energie, Tropfen- bzw. Aggregatgröße, etc.) mit der dazugehörigen Übergangstemperatur und -breite, als auch eine mikrokanonische Analyse sowie die Barrieren der Freien Energie. Für semiflexible Polymere wird insbesondere der Einfluss von Steifigkeit auf die resultierende Struktur der Aggregate untersucht, die von amorphen Kugeln für flexible Polymere bis hin zu verdrehten Bündeln für steifere Polymere reichen.
Ein weiterer Fokus liegt auf der Untersuchung von Übereinstimmungen zwischen den generischen Mechanismen in Kondensation und Aggregation: dem Übergang zwischen einer homogenen Phase und einer inhomogenen (gemischten) Phase. Auf diesem Niveau kann man Polymeraggregation als Kondensation von ausgedehnten Objekten verstehen. Dies zeigt sich vor allem in dem Skalierungsverhalten von kanonischen und mikrokanonischen Observablen, insbesondere an einem unerwarteten aber konsistenten Bereich für mittelgroße (mesoskopische) Systemgrößen.
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Biomass to Biofuel : Syngas Cleaning and Biomass FeedstockSadegh-Vaziri, Ramiar January 2017 (has links)
This thesis builds around the idea of a biofuel production process that is comprised of biomass production, biomass gasification, gas cleaning, and fuel production. In this work, we specifically looked into H2S removal as a part of cleaning the producer gas and flocculation of microalgae which is involved in the harvesting of microalgae after biomass production. One of the impurities to remove from the producer gas is hydrogen sulfide which can be removed by using a packed bed of zinc oxide. Despite the regular use, it was only recently shown that during reaction with H2S, nano-size particles of ZnO exhibit void formation and outward growth. In this work, a micro-scale model was introduced to describe the void formation and outward growth. On the macro-scale, the simulations captured pore clogging of pellets due to the outward growth. The pore clogging prevents the full conversion of pellets and consequently leads to shorter breakthrough times of beds. The second problem investigated here deals with the flocculation of microalgae. Microalgae is produced in relatively low concentrations in the incubator liquid medium and during the harvesting, the concentration is increased to an acceptable level. The harvesting process includes a flocculation followed by a filtration or centrifuge unit. During flocculation, microalgae are stimulated to aggregate and form clusters. The experiments showed that the mean size of clusters formed during flocculation increases with time to a maximum and then starts decreasing, resulting in an overshoot in the mean size profile. The size of clusters influence the efficiency of the afterward filtration or centrifuge, thus it is of interest to carefully track the size evolution of clusters, making the studying of overshoot a crucial research topic. In this work, the possible mechanisms behind this overshoot were investigated. / <p>QC 20170330</p>
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