Apesar de extensa investigação das modificações oxidativas irreversíveis sofridas pelas proteínas in vitro e in vivo, os produtos formados pela oxidação de resíduos de triptofano ainda permanecem apenas parcialmente conhecidos. Recentemente, nosso grupo caracterizou uma ligação cruzada de ditriptofano produzida pela recombinação de radicais hSOD1-triptofanila gerados pelo ataque do radical carbonato produzido durante a atividade peroxidásica da enzima superóxido dismutase humana (hSOD1). Neste trabalho, examinamos se a ligação ditriptofano pode ser formada em outras proteínas, além da hSOD1 e por outros oxidantes, além do radical carbonato. A lisozima da clara do ovo e a beta cristalino bovina foram utilizadas como alvos de oxidação. A lisozima foi utilizada por ser uma enzima pequena (129 aminoácidos) e de estrutura bem conhecida, contendo seis resíduos de Trp. Os resultados mostraram que o radical carbonato, gerado enzimatica ou fotoliticamente, promove a oxidação, dimerização e inativação da lisozima. Os principais produtos de oxidação caracterizados por análise de nano-ESI-Q-TOF-MS/MS foram hidroxi-triptofano e N-formilquinurenina juntamente com um dímero de lisozima (lisozima-Trp28-Trp28-lisozima) e um hetero dímero lisozima-hSOD1 (lisozima-Trp28-Trp32-hSOD1), ambos ligados por uma ligação ditriptofano. Também demonstramos que a irradiação da lisozima com luz UVC leva à formação do dímero lisozima-Trp28-Trp28-lisozima. Em consequência, resolvemos tratar a beta cristalino bovina com radical carbonato gerado fotoliticamente ou com luz UVC, e a proteína também sofreu oxidação, dimerização e agregação. Os principais produtos de oxidação caracterizados por nano-ESI-Q-TOF-MS/MS foram hidroxi-triptofano, N-formilquinurenina, DOPA e um dímero de beta cristalino (βB2-Trp151-Trp151-βB2). A irradiação com luz UVC também levou à formação de um dímero intra-cadeia, caracterizado como βA2-Trp78-Trp81. Quando a beta cristalino foi irradiada com um simulador de luz solar (UVA e UVB) também foi possível observar um dímero, caracterizado como βA2-Trp150-Trp150-βA2. A presença de produtos de oxidação de resíduos de Trp, dentre eles a ligação cruzada ditritpofano, também foi avaliada in vivo, utilizando o cristalino de pacientes que foram submetidos a cirurgia para remoção de catarata. Beta, alfa e gama cristalino foram as principais proteínas identificadas nas frações solúvel e insolúvel do cristalino. A principal modificação pós-traducionais identificada foi deamidação. Um alto conteúdo de resíduos de metionina e triptofano oxidados foram identificados nas proteínas presentes na fração insolúvel. Os principais produtos de oxidação de Trp identificados por nano-ESI-Q-TOF-MS/MS foram quinurenina e N-formilquinurenina. A presença de dímeros covalentes no cristalino com catarata foi confirmada por análises de massas. A completa caracterização desses dímeros (βB1-Trp127-Trp127-βB1 e βB1-Trp193-Trp193-βB1) confirmou que as cadeias polipeptídicas foram ligadas por uma ligação ditriptofano. Em síntese, nossos dados demonstraram que o radical carbonato e a luz UV podem produzir dímeros de ditriptofano em diferentes proteínas. Também, a presença da ligação cruzada de ditriptofano in vivo (catarata humana) foi pela primeira vez detectada. / Despite extensive investigation of irreversible oxidative modifications suffered by proteins in vitro and in vivo, the products formed by oxidation of tryptophan residues remain partially characterized. Our group recently described a ditryptophan cross-link produced by recombination of hSOD1-tryptophanyl radicals generated by attack of the carbonate radical produced during the peroxidase activity of the human superoxide dismutase (hSOD1) enzyme. Here, we examine whether the ditryptophan cross-link can be produced in others proteins besides the hSOD1 and by other oxidants, in addition to the carbonate radical. The egg white lysozyme and bovine beta crystalline were used as targets. Lysozyme was used because it is a small enzyme (129 amino acids) with a well-known structure, containing six Trp residues. The results showed that the carbonate radical, generated enzymatically or photolytically, promotes lysozyme oxidation, inactivation and dimerization. The major oxidation products characterized by nano-ESI-Q-TOF-MS/MS analysis were hydroxy-tryptophan and N-formylkynurenine together with a dimer of lysozyme (lysozyme-Trp28-Trp28-lysozyme) and a hetero dimer hSOD1-lysozyme (lysozyme-Trp28-Trp32-hSOD1), both bound by a ditryptophan cross-link. Also, it was demonstrated that lysozyme irradiation with UVC light leads to the formation of the dimer lysozyme-Trp28-Trp28-lysozyme. In view of these results, we decided to treat beta crystalline bovine with photolytically generated carbonate radical and UVC. Beta crystalline also suffered oxidation, dimerization and aggregation. The major oxidation products characterized were hydroxy-tryptophan, N-formylkynurenine, DOPA and a beta crystalline dimer (βB2-Trp151-Trp151-βB2) by nano-ESI-Q-TOF-MS/MS. Irradiation with UVC light also led to the formation of an intra-chain dimer, which was characterized as βA2-Trp78-Trp81. When beta crystalline was irradiated with a solar simulator (UVA and UVB), it was also possible to observe a dimer which was characterized as βA2-Trp150-Trp150-βA2. The presence of oxidized tryptophan products, including the ditryptophan cross-link, was also evaluated in vivo in the lenses of patients submitted to cataract removal. Beta, alpha and gamma crystalline were the main proteins identified in soluble and insoluble fractions of the lenses. The main post translational modification identified was deamidation. A high content of oxidized methionine and tryptophan residues were identified in proteins present in the insoluble fraction. The main tryptophan oxidation products identified by nano-ESI-Q-TOF-MS/MS were kynurenine and N-formylkynurenine. The presence of covalent dimers in the lenses with cataract was demonstrated by mass analysis. Full MS/MS characterization of the dimers βB1-Trp127-Trp127-βB1 and βB1-Trp193-Trp193-βB1 confirmed that they were linked by a ditryptophan bond. In summary, our data demonstrate that the carbonate radical and UV light can produce ditryptophan dimers in different proteins. Also, the presence of the ditryptophan cross-link was first detected in vivo (human cataract).

Agrotóxicos são frequentemente relatados na literatura em contaminações de ambientes aquáticos, sendo provenientes do lixiviamento de solos, do descarte inadequado de embalagens agrícolas, dentre outros. Nesse contexto, os processos oxidativos avançados (POA) têm sido estudados como alternativa para o tratamento desses compostos em meio aquoso. Os POA consistem na oxidação de compostos orgânicos pela formação de radicais livres com alto poder oxidante. Diante desses aspectos, o presente trabalho teve como proposta o estudo da degradação dos herbicidas hexazinona e diuron, aliada ao planejamento experimental do tipo composto central, a fim de otimizar o processo de degradação. Para tanto, realizaram-se as degradações através dos processos H2O2/UV e foto-Fenton em concentrações iniciais próximas a 7 e 20 mg L-1 para a hexazinona e diuron, respectivamente. Os experimentos foram conduzidos em um reator fotoquímico (200 mL a 25 °C) com aplicação de irradiação UV, proveniente de uma lâmpada de Hg (degradação H2O2/UV) e de luz negra (degradação foto-Fenton). No processo via H2O2/UV foram avaliadas a concentração de H2O2 (0,65 a 13,34 mmol L-1) e pH (2,77 a 11,23), e para o processo foto-Feton avaliou-se a concentração de H2O2 (0,09 a 29,1 mmol L-1) e Fe (II) (0,01 a 0,92 mmol L-1). Como variável independente, utilizou-se a remoção de Carbono Orgânico Total (COT) nos dois casos. A eficiência na degradação dos herbicidas foi determinada através das técnicas de Espectroscopia UV-Vis, Cromatografia Líquida de Alta Eficiência (HPLC/UV), remoção de Carbono Orgânico Total e Cromatografia de Íons. A análise dos modelos matemáticos, obtidos a partir dos planejamentos, possibilitou a avaliação da influência das variáveis, determinando as melhores condições. No processo H2O2/UV observou-se que a influência da variável H2O2 é maior comparada ao pH. A melhor condição de degradação apresentou concentrações de H2O2 iguais a 7 mmol L-1 e valores de pH de 2,8. Como resultado, obteve-se 96 % de remoção do COT e a não detecção de ambos os herbicidas após 2 minutos de reação. No processo foto-Fenton, a realização de três planejamentos foi necessária para a otimização do sistema. Primeiro e segundo planejamentos indicaram a melhor condição, enquanto que o terceiro planejamento revelou que concentrações elevadas de H2O2 e Fe2+ não garantem maior eficiência. As condições ideais encontradas foram iguais a 0,291 mmol L-1 para o Fe2+ e 2,91 mmol L-1 para o H2O2, obtendo-se 76 % de remoção do COT e não detecção dos herbicidas nos primeiros minutos de degradação. Em ambas as degradações não foi possível observar diferenças em relação a cinética de degradação para os herbicidas. A degradação via H2O2/UV mostrou melhor desempenho, pois apresentou maior taxa de mineralização além de não necessitar o emprego do íon Fe(II). / Pesticides are often reported in the literature on aquatic environments contamination, being a result of soil leaching, improper disposal of agricultural packages, among others. In this context, advanced oxidation processes (AOP) have been studied as an alternative to treat these compounds in aqueous medium. The AOP consist in the oxidation of organic compounds through the formation of free radicals with a high oxidizing power. Considering these aspects, the present work aimed to study degradation of the herbicides diuron and hexazinone, coupled with an experimental setup as central compost, in order to optimize the degradation process. For this purpose, the degradation was carried out by H2O2/UV and photo-Fenton processes at initial concentrations close to 7 and 20 mg L-1 to hexazinone and diuron, respectively. The experiments were conducted in a photochemical reactor (200 mL at 25 ° C) with application of UV irradiation from Hg lamp (H2O2/UV degradation) and black-light (photo-Fenton degradation). In the H2O2/UV process, the concentrations of H2O2 (0.65 to 13.34 mmol L-1) and pH (2.77 to 11.23) were evaluated, and in the photo-Fenton process, the concentrations of H2O2 (0.09 to 29.1 mmol L-1) and Fe (II) (0.01 to 0.92 mmol L-1) were analyzed. In both processes, the removal of Total Organic Carbon (TOC) was used as an independent variable. The efficiency of the herbicide degradation was determined by UV/Vis spectroscopy, high performance liquid chromatography (HPLC/UV), removal of the Total Organic Carbon and ion chromatography. The analysis of the mathematical models obtained from the experimental setup allowed the evaluation of the variables influence ,determining the best conditions. The H2O2/UV process showed that the influence of the H2O2 is higher compared to that of the pH. The best degradation condition presented H2O2 concentrations of 7 mmol L-1 and a pH value of 2.8. As a result, 96% TOC removal was observed, and none of the herbicides were detected after 2 minutes of reaction. In the photo-Fenton process, the conduction of three setups was necessary to confirm the best condition. The first and second setups indicated the best condition, while the third one showed that high concentrations of H2O2 and Fe2+ do not guarantee a greater degradation efficiency. Ideal conditions were equal to 0.291 mmol L-1 for Fe2+ and 2.91 mmol L-1 for H2O2, where 76% of TOC removal was obtained and also no herbicides detection was seen in the early minutes of degradation. In both processes, it was not possible to observe differences in the degradation kinetics of the herbicides. The H2O2/UV degradation showed better performance because of a greater mineralization rate besides not requiring the ion Fe (II) use.

Patients of Hutchinson-Gilford Progeria Syndrome (HGPS) display a rate of aging up to ten times that of normal human populations. It might be expected that HGPS cells would have a decreased ability to repair DNA damage through the cell cycle as compared to normal cells such as those of the BJ cell line since DNA damage accumulation is a hallmark phenotype of aging. On earth, we are exposed to far more ultraviolet-B (UV-B, 280-315 nm) and UV-A (315-400 nm) than UV-C (100-280 nm) radiation, since the latter is filtered-out by the atmospheric ozone layer. The relative sensitivity of prematurely aging HGPS cells to UV-B irradiation is unknown. It was hypothesized that the normal fibroblast cell line (BJ) would exhibit a higher rate of DNA repair and a higher level of cell viability after exposure to ultraviolet radiation than would be observed with the HGPS cells, and that these differences would be greater as the HGPS cells age in culture. A Cell-Titer Blue Viability Assay (Promega) was used to determine the effect of UV-B and UV-C on metabolic activity, an indicator for cell viability, in HGPS, BJ, and A549 (a human lung carcinoma) cells. A translesion DNA synthesis protein, pol-η, and several other DNA transcription and repair-related proteins also were hypothesized to be altered in the HGPS cell line, both before and after UV-induced DNA damage, as compared to the BJ cell line. Western blotting was used to monitor these proteins in BJ and HGPS cells following UV-C exposure. No differences in short-term viability were observed between BJ and HGPS cells, reflecting similarities in their repair abilities on the cellular level; however, there were significant differences in long-term viability. Enzyme Linked Immunosorbant Assays (ELISA) revealed a significant difference in DNA repair at the molecular level. Moreover, Western blotting revealed differences in the amounts of several repair-related proteins following UV exposure, including pol-η, an important trans-lesion synthesis protein. Although the difference in DNA repair did not appear at the cellular level, it is apparent that HGPS cells show a greater sensitivity to both UV-B and UV-C irradiation as compared to normal BJ fibroblasts and A549 carcinoma cells.

Cette thèse a été réalisée entre 2014 et 2017 en vue de la préparation d'une mission probatoire du projet Imageur de Fresnel permettant de réaliser des observations astronomiques dans le domaine ultraviolet (UV). Il s'agit d'un modèle de télescope spatial diffractif imaginé et développé principalement par Laurent Koechlin depuis 2004. Son optique primaire est une grille de Fresnel, optique très légère (de l'ordre du kilogramme) parfaitement adaptée pour servir à une mission d'observation spatiale de très grande envergure, utilisant le principe des réseaux zonés de Fresnel. Afin de prouver la faisabilité d'une mission probatoire sur la Station Spatiale Internationale (ISS) fonctionnant dans l'ultraviolet, l'instrument doit d'abord faire la preuve de ses performances au sol. Le premier objectif a donc été de concevoir un nouveau prototype sol adapté à l'observation dans l'ultraviolet. Cela a nécessité la définition d'une nouvelle configuration de l'ensemble du système imageur, ainsi que la réalisation de nouvelles optiques, et en particulier celle du correcteur du chromatisme induit par la grille d'un nouveau genre. Il s'agit d'un miroir de Fresnel blazé concave, qui a été réalisé pour la première fois avec succès. Le second a été de perfectionner le modèle de grille de Fresnel, afin d'en améliorer ses qualités de haut contraste en conservant sa résolution maximale. Cela s'est fait par la modification des barreaux de maintien des anneaux, ainsi que par l'application d'une apodisation adaptée à cette optique particulière. La nouvelle grille du prototype d'une largeur de 65 mm seulement, permet théoriquement d'atteindre une dynamique de 10^6 à 15 resels (éléments de résolution), probablement meilleure avec la grille qui sera utilisée pour la mission probatoire, et bien supérieure encore avec les grilles de grandes dimensions si un jour elles sont utilisées pour l'astrophysique UV. / This Ph.D. thesis work was made between 2014 and 2017, in order to prepare a probatory mission of the Fresnel Imager for astronomical observations in the ultraviolet (UV) domain. The Fresnel Imager is a concept of diffractive telescope created and developed mainly by Laurent Koechlin since 2004. Its primary optics is a Fresnel grid: a very light weight optics (in the range of kilograms) using the principle of Fresnel zone plates. Fresnel arrays are adapted to large apertures in space. In order to prove the feasibility of a probatory mission on the International Space Station (ISS), the instrument has first to prove its performances on the ground. The initial goal of my thesis was to conceive a new prototype adapted to UV. This required a new configuration of the entire imaging system, as well as the realization of new optics, particularly those correcting the chromatism induced by that new kind of diffractive imaging. This chromatic corrector is a concave blazed Fresnel mirror, which has been successfully realized for the first time. The second goal was to improve the design of the Fresnel grid, in order to enhance its high contrast performances while preserving its diffraction-limited resolution. This was obtained by modifying the setup which holds the rings in place, as well as by the application of an apodization adapted to this particular optics. The new Fresnel grid in the prototype is 65 mm x 65 mm only, but its point spread function reaches a dynamic range of 10^6 at 15 resels (resolution element) from center. This will be further improved when a slightly larger array planned for the probatory mission on the ISS. Later, space missions could feature Fresnel grids several meters in size, yielding diffraction-limited images in the UV.

Ces travaux de thèse se positionnent au cœur des problématiques du patrimoine et notamment de la conservation préventive des œuvres d'art. Pour apporter aux musées une solution à faible coût capable de répondre aux exigences fixées dans le cadre de la protection du patrimoine culturel, nous avons développé de nouveaux vitrages polymères permettant de protéger les œuvres sans en altérer leur perception par le public. Ces matériaux pourront, à terme, tenter de remplacer les vitrages de type poly(méthacrylate de méthyle) (PMMA), très largement utilisés, mais dont le principal inconvénient est sa grande sensibilité à la rayure et aux solvants de nettoyage.Ces matériaux ont été synthétisés en combinant, en différentes proportions, deux polymères, le PMMA et le polycarbonate de CR39® (PCR39®) dans une architecture de réseaux interpénétrés de polymères (RIP). Cette association a permis d'obtenir des nouveaux matériaux transparents (T%>95%) dont la résistance à la rayure et aux solvants est considérablement améliorée comparativement à celle du PMMA. Leurs vieillissements lors de leur exposition à la lumière (UV) artificielle ou naturelle ont été étudiés. Afin d'améliorer leurs propriétés d'anti-rayure, nous avons réticulé sélectivement la phase PMMA avec des nanoparticules de silice fonctionnalisées, ce qui conduit à la réalisation d'un nouveau RIP hybride. Enfin, dans l'optique d'obtenir un matériau intelligent, nous avons étudié la possibilité d'intégrer au sein même de ces vitrages, un capteur de rayonnements UV afin de pouvoir alerter de leur présence. Le PCR39® ayant les propriétés en adéquation avec le développement d'un tel capteur, nous avons ensuite cherché à augmenter la sensibilité du matériau. Pour cela, des composés fluorescents, organique, d'une part et inorganiques d'autre part (nanoparticules d'oxyde de gadolinium dopées soit au terbium soit à l'europium) ont été intégrés au sein des RIP. / This work position itself on a primary issue of heritage and more precisely on the preventive conservation of artworks. To provide a low cost solution to museums, capable of meeting the requirements set for the cultural heritage protection, we have developed new polymer glazing to protect works without altering their perception by the public. These materials will eventually be able to replace poly(methyl methacrylate) (PMMA) glazing, which are widely used but have high sensitivity to scratching and cleaning solvents as mains drawbacks.These materials were synthesized by combining, with various proportions, two polymers, PMMA and polycarbonate of CR39® (PCR39®), into an interpenetrating polymer network (IPN) architecture. This association generated new transparent materials (T% > 95%) of which resistance toward scratching and solvents is considerably improved compared to PMMA's one. Their ageing have been studied when exposed to artificial or natural light (UV). To improve their scratch resistance, we selectively crosslinked the PMMA phase with functionalized silica nanoparticles, leading to the formation of a new hybrid IPN. Finally, in order to obtain a smart material, we studied the possibility of integrating within these glazing, a UV sensor to warn of their presence. The PCR39® having properties consistent with the development of such a sensor, we tried to increase the sensitivity of the material. For that purpose, first an organic fluorescent compounds, and secondly an inorganic one (gadolinium oxide nanoparticles doped with terbium or europium) were incorporated in the IPN.

Interferentes endócrinos são substâncias que podem causar danos ao meio ambiente e à saúde humana. Estes compostos são usados na produção de vários produtos de uso diário, porém não são completamente eliminados dos efluentes industriais, contribuindo para a contaminação de diversas matrizes ambientais. Nesse contexto, os processos eletroquímicos se destacam com boas eficácias no tratamento de diversos tipos de efluentes. No entanto, uma melhor compreensão do processo é importante para estabelecer tratamentos de efluentes que possam ser mais amplamente aplicados. Esta tese aborda a degradação do metil parabeno (MeP), em diferentes efluentes sintéticos, via eletrólise com ânodos de misturas de óxidos metálicos comercial (MOM-Cl2) e diamante dopado com boro (DDB). Ainda mais importante, o MeP é utilizado como uma molécula modelo para o compreender os efeitos da irradiação de luz UV e de ultrassom (US) durante a eletrólise. É importante lembrar que a composição da matriz deve afetar a eficiência do processo e, assim, também se utiliza o MeP como modelo para entender os possíveis efeitos interferentes de outros compostos orgânicos na sua oxidação. Finalmente, se estuda um efluente simulado da indústria cosmética para melhor compreender os efeitos da eletrólise e da irradiação de US em uma matriz complexa. Foram observadas grandes diferenças entre as eficiências dos tratamentos com MOM-Cl2 e DDB, principalmente com relação ao mecanismo de degradação do MeP. Nos estudos com MOM-Cl2, muitas vezes o efluente final era mais complexo que o inicial, devido à formação de produto sólido, reações de polimerização e alto efeito de interferência quando dois compostos são tratados simultaneamente. Com o DDB, o processo de mineralização é muito mais favorecido, independentemente da complexidade do efluente. O efeito de interferência ganha significância na presença de altas concentrações de um segundo composto orgânico, mas a completa mineralização ainda foi atingida. A irradiação de UV e US representou melhoras no processo de remoção de matéria orgânica, principalmente com relação aos produtos do MeP. Foi observado que surfactantes podem interferir negativamente na degradação de compostos orgânicos devido a formação de micelas. Diante de alto conteúdo micelar a eficiência do processo diminui, porém 90% de mineralização do efluente foi alcançada. A irradiação de US ajuda significativamente no aspecto físico do efluente tratado, sendo possível obter a transformação de emulsões, com alta turbidez e material particulado, em soluções completamente límpidas e mineralizadas. / Endocrine disrupting chemicals (EDCs) are responsible for causing adverse effects on the environment and on human health. Those substances are extensively used in several costumer products, such as pharmaceuticals and cosmetics. However, several EDCs are not completely removed from industrial wastewaters by the most common treatments, contributing on the environmental contamination. In this context, electrochemical processes can be highlighted due to its efficacy presented on the treatment of numerous types of wastewater. Nevertheless, better understanding of the process is important in order to design wastewater treatments that can be widely applied. Hence, this thesis focus on the degradation of methyl paraben (MeP), in different synthetic wastewater, by electrolysis with mixed metal oxides commercial (MMO-Cl2) and boron-doped anode (BDD) anodes. More importantly, MeP is used as a model molecule to understand the effects of the irradiation of UV light and of ultrasound (US) during the electrolysis. It is important to consider that matrix composition must affect the process efficiency. Hence, MeP is also used as model to shed a light on the possible interfering effects of other organics on its oxidation. Finally, a simulated wastewater from cosmetic industry is studied in order to better comprehend the effects of electrolysis and of ultrasound on complex matrix. Great differences were observed on the efficiency of treatments using MMO-Cl2 and BDD, mostly regarding the mechanism of MeP oxidation. With MMO-Cl2, the treated wastewater was often more complex than the initial one, due to the formation of solid product, polymerization reaction and great interfering effect when two compounds are simultaneously treated. Conversely, using BDD, the mineralization process is favored, regardless the complexity of the matrix. Interfering effect on MeP oxidation is relevant under the presence of huge concentrations of a second organic compound, though the complete mineralization is attained. Irradiation of UV and of US resulted in better removals of organic matter, mainly with respect to MeP products. It was observed that surfactants may interfere on the degradation of organic compounds due to the formation of micelles. The efficiency of the process decreases under high micelles content, however 90% of mineralization was achieved. Irradiation of ultrasound notably improves the physical aspect of the treated wastewater, assisting on the transformation of emulsions, with great turbidity and amount of particles, in clear non-organic solutions.

In this work approaches for new detection system development for an Analytical Ultracentrifuge (AUC) were explored. Unlike its counterpart in chromatography fractionation techniques, the use of a Multidetection system for AUC has not yet been implemented to full extent despite its potential benefit. In this study we tried to couple existing fundamental spectroscopic and scattering techniques that are used in day to day science as tool for extracting analyte information. Trials were performed for adapting Raman, Light scattering and UV/Vis (with possibility to work with the whole range of wavelengths) to AUC. Conclusions were drawn for Raman and Light scattering to be a possible detection system for AUC, while the development for a fast fiber optics based multiwavelength detector was completed. The multiwavelength detector demonstrated the capability of data generation matching the literature and reference measurement data and faster data collection than that of the commercial instrument.<br><br> It became obvious that with the generation of data in 3-D space in the UV/Vis detection system, the user can select the wavelength for the evaluation of experimental results as the data set contains the whole range of information from UV/Vis wavelength. The detector showed the data generation with much faster speed unlike the commercial instruments. The advantage of fast data generation was exemplified with the evaluation of data for a mixture of three colloids. These data were in conformity with measurement results from normal radial experiments and without significant diffusion broadening. Thus conclusions were drawn that with our designed Multiwavelength detector, meaningful data in 3-D space can be collected with much faster speed of data generation. / In der vorliegenden Arbeit wurde die Entwicklung neuer Detektoren für die Analytische Ultrazentrifugation (AUZ) untersucht und vorangetrieben. Im Gegensatz zu chromatgraphischen. Fraktionierungsmethoden werden Multidetektionssysteme bis heute nicht in der AUZ eingesetzt. Hier wird die Möglichkeit geprüft, bekannte spektroskopische sowie Streumethoden simultan zur Probenanalyse in der AUZ einzusetzen mit dem Ziel, simultan komplimentäre Informationen über die Probe zu erhalten. So wurde versucht, Raman- und UV/VIS-Spektroskopie (letztere mit der Möglichkeit, das gesamte Wellenlängenspektrum auszunutzen) und statische Lichtstreuung zu kombinieren, um das Analytverhalten während des Ultrazentrifugationsexperimentes zu untersuchen. Es wurden zum einen die Ramanspektroskopie als Detektionssystem für chemische Funktionalität in der AUZ geprüft und zum anderen gezeigt, daß die statische Kleinwinkel Lichtstreuung als direkter Molmassendetektor für den Einsatz in der AUZ geeignet erscheint. Zum anderen wurde die Entwicklung eines Multi-Wellenlängen-UV/VIS-Detektors abgeschlossen, um seine Eignung für den Einsatz in der AUZ und die damit verbundene Möglichkeiten der schnelleren und umfassenderen Datenerzeugung gegenüber kommerziellen Geräten zu zeigen. Dieser Multiwellendetektor liefert anstelle eines Absorptionswertes für jede radiale Position in der Messzelle direkt ein ganzes UV-Vis Spektrum und erzeugt eine zusaetzliche Dimension der Messdaten, was die Möglichkeiten der Analyse von komplexen Systemen mit multiplen Chromophoren, Teilchengrößenbestimmung über Wellenlängenabhängigkeit der Trübung oder auch der Datenmittelung enorm vergrößert. Desweiteren erlaubt der Detektor die Anwendung von Geschwindigkeitsprofilen zur Analyse extrem polydisperser Systeme. Die Entwicklung des Detektors beruht auf einem auf Linsen basierenden System mit modularem Aufbau. Dabei war die sorgfältige Ausrichtung des optischen Systems ein essentieller Punkt, um seine Eignung zu überprüfen zu können. An einer Mischung von drei Kolloiden, Halbleiternanopartikeln sowie Proteinen und deren Mischungen ist es hier gelungen, die erfolgreiche Entwicklung des UV/VIS-Detektors zu demonstrieren: Die Daten konnten schneller und mit wesentlich mehr Informationsgehalt, als auf allen kommerziellen Geräten generiert werden. Die Sedimentationskoeffizientenverteilungen stimmen dabei mit denen aus herkömmlichen Sedimentationsgeschwindigkeitsexperimenten überein, unterliegen jedoch nicht einer signifikanten diffusionsbedingten Verbreiterung. Es ist in dieser Arbeit somit gelungen, zum einen die Lichtstreuung als aussichtsreiche Methode für ein Detektorsystem in der AUZ aufzuzeigen, und zum anderen einen Multi-Wellenlängen-UV/VIS-Detektor zu entwickeln, der eine Datenerzeugung von bislang noch nicht erreichter Schnelligkeit im dreidimensionalen Raum ermöglicht.

Aluminum Gallium Nitride (AlGaN) / Gallium Nitride (GaN) based deep ultraviolet (DUV) light emitting didoes (LEDs) with emission wavelengths between 200-280 nm enable key emerging technologies such as water/air purification and sterilization, covert communications and portable bio-agent detection/identification systems for homeland security, and surface and medical device sterilization. These devices produce a large amount of undesired heat due to low quantum efficiencies in converting electrical input to optical output. These low efficiencies are attributed to difficulties in the growth&doping of AlₓGa₁₋ₓN materials and UV absorbing substrates leading to excessive joule heating, which leads to device degradation and a spectral shift in the emission wavelength. With this regard, effective thermal management in these devices depends on the removal of this heat and reduction of the junction temperature. This is achieved by decreasing the package thermal resistance from junction-to-air with cost-effective solutions. The use of heat sinks, thermal interface materials, and high conductivity heat spreaders is instrumental in the reduction of the overall junction-to-air thermal resistance. This thesis work focuses on thermal modeling of flip-chip packaged deep UV LEDs to gain a better understanding of the heat propagation through these devices as well as the package parameters that have the biggest contributions to reducing the overall thermal resistance. A parametric study focusing on components of a lead frame package is presented to ascertain the thermal impacts of various package layers including contact metallizations, thermal spreading sub-mounts, and thermal interface materials. In addition the use of alternative thermal interface materials such as phase change materials and liquid metals is investigated experimentally.

Um das Verhalten von Actiniden im Menschen (Stoffwechsel), in geologischen und in biologischen Systemen vorherzusagen, ist es erforderlich deren Speziation genau zu kennen. Zur Bestimmung dieser wird das chemische Verhalten des Urans hinsichtlich Komplexbildungsreaktionen und Redoxreaktionen in Modellsystemen untersucht. Anhand der gewonnenen thermodynamischen Konstanten und dem Redoxverhalten können Risikoabschätzungen für das jeweilige untersuchte System getroffen werden. Das umweltrelevante Uran(IV)-Uran(VI)-Redoxsystem besitzt mit der metastabilen fünfwertigen Oxidationsstufe einen zumeist kurzlebigen Zwischenzustand. Innerhalb dieser Arbeit gelang es erstmalig die Uran(V)-Fluoreszenz mittels laserspektroskopischer Methoden nach zu weisen. Beispielsweise konnte das Bandenmaximum von aquatischem Uranyl(V) im perchlorhaltigem Medium (λex = 255 nm) mit 440 nm, bei einer Fluoreszenzlebensdauer von 1,10 ± 0,02 µs bestimmt werden. Die fluoreszenzspektroskopische Untersuchung eines aquatischen [U(V)O2(CO3)3]5--Komplexes (λex= 255 nm und 408 nm) zeigte bei Raumtemperatur keine Fluoreszenz. Durch Anwendung der Tieftemperaturtechnik wurden bekannte Quencheffekte des Carbonats unterdrückt, so dass bei beiden Anregungswellenlängen ein für Uran(V) typisches Fluoreszenzspektrum im Bereich von 375 nm bis 450 nm, mit Bandenmaxima bei 401,5 nm (λex = 255 nm) und 413,0 nm (λex = 408 nm) detektiert werden konnte. Darüber hinaus konnte bei 153 K (λex = 255 nm) eine Fluoreszenzlebensdauer von 120 ± 0,1 µs bestimmt werden. Untersetzt wurden diese fluoreszenzspektroskopischen Nachweise durch mikroskopische Studien verschiedener Uran(IV)-Festphasen (Uraninit…UO2, Uran(IV) Tetrachlorid…UCl4) und einer sulfathaltigen Uran(IV)-Lösung (UIVSO4). Diese wurden durch kontinuierliche Sauerstoffzufuhr zu Uran(VI) oxidiert. Die ablaufende Oxidation wurde mit dem konfokalen Laser Scanning Mikroskop (CLSM) verfolgt, wobei die Proben mit einer Wellenlänge von 408 nm zur Fluoreszenz angeregt wurden. Die auftretenden Bandenmaxima bei 445,5 nm (UO2), bei 445,5 nm (UCl4) und bei 440,0 nm (UIVSO4) konnten eindeutig der Uran(V)-Fluoreszenz zugeordnet werden. Zur Bestimmung thermodynamischer Konstanten mit Hilfe der Tieftemperaturfluoreszenz wurde zunächst der Einfluss der Temperatur auf das Fluoreszenzverhalten des freien Uranyl(VI)-Ions näher betrachtet. Es zeigte sich, dass mit Erwärmung der Probe (T>298 K) die Fluoreszenzlebensdauer von 1,88 µs (298 K) deutlich absinkt. Die Fluoreszenzintensität verringerte sich dabei um 2,3 % pro 1 K zwischen 273 K und 313 K. Im Gegensatz dazu, steigt die Fluoreszenzlebensdauer um das 150-fache auf 257,9 µs bei einer Verminderung der Temperatur (T <298 K) auf 153 K. Das weitere Absenken der Temperatur (T <153 K) zeigte keinen Einfluss auf die Fluoreszenzlebensdauer. Die Lage der Hauptemissionsbanden des freien Uranyl(VI)-Ions (488,0 nm, 509,4 nm, 532,4 nm, 558,0 nm, 586,0 nm) zeigte bei diesen Untersuchungen keine temperaturabhängige Verschiebung. Die Validierung der Tieftemperaturtechnik zur Bestimmung thermodynamischer Konstanten mittels zeitaufgelöster laserinduzierten Fluoreszenzspektroskopie erfolgte anhand des Uran(VI)-Citrat-Systems. Im Gegensatz zu bisherigen fluoreszenzspektroskopischen Betrachtungen bei Raumtemperatur wurde das Fluoreszenzsignal bei tiefen Temperaturen mit einsetzender Komplexierung nicht gequencht, woraus die Ausprägung einer gut interpretierbaren Fluoreszenz resultierte. Die Analyse der spektralen Daten mit SPECFIT ergaben mit log β101 = 7,24 ± 0,16 für den [UO2(Cit)]--Komplex und log β202 = 18,90 ± 0,26 für den [(UO2)2(Cit)2]2 -Komplex exakt die in der Literatur angegebenen Stabilitätskonstanten. Zudem konnten Einzelkomponentenspektren mit Bandenmaxima bei 475,3 nm, 591,8 nm, 513,5 nm, 537,0 nm und 561,9 nm für den 1:0:1-Komplex und 483,6 nm, 502,7 nm, 524,5 nm, 548,1 nm und 574,0 nm für den 2:0:2-Komplex und Fluoreszenzlebensdauern von 79 ± 15 µs (1:0:1) und 10 ± 3 µs (2:0:2) bestimmt werden. Zur Modellkomplexierung des Uran-Citrat-Systems wurde in dieser Arbeit auch das Komplexbildungsverhalten von U(IV) in Gegenwart von Citronensäure untersucht. Hierbei wurden über den gesamten pH-Wertbereich gelöste Uran-Citrat-Spezies spektroskopisch nachgewiesen und die Stabilitätskonstanten sowie die Einzelkomponentenspektren für die neu gebildeten Uran(IV) und (VI)-Spezies bestimmt. Für die neu gebildeten Citrat-Komplexe des sechswertigen Urans wurden Komplexbildungskonstanten von log β203 = 22,67 ± 0,34 ([(UO2)2(Cit)3]5-) und log β103 = 12,35 ± 0,22 ([UO2(Cit)3]7-) und für die Komplexe des vierwertigen Urans von log β1-21 = -9,74 ± 0,23 ([U(OH)2Cit]-) und log β1 31 = -20,36 ± 0,22 ([U(OH)3Cit]2-) bestimmt. Untersuchungen zum Redoxverhalten von Uran in Gegenwart von Citronensäure zeigten unter aeroben und anaeroben Versuchsbedingungen eine photochemische Reduktion vom U(VI) zu U(IV), welche spektroskopisch nachgewiesen werden konnte. Dabei zeigt speziell die Reaktion unter oxidierenden Bedingungen, welchen großen Einfluss vor allem organischen Liganden auf das chemische Verhalten des Urans haben können. Sowohl die Reduktion unter O2- als auch die unter N2-Atmosphäre, weisen ein Maximum bei einem pH Wert von 3,5 bis 4 auf. Unter anaeroben Bedingungen reduziert die Citronensäure mit ca. 66 %, 14 % mehr Uran(VI) zu Uran(IV) als unter anaeroben Bedingungen mit ca. 52 %. Ab einem pH-Wert von 7 konnte eine Reduktion nur unter sauerstofffreien Bedingungen festgestellt werden. Die Wechselwirkung von U(VI) in Gegenwart von Glucose wurde hinsichtlich Reduktion und Komplexierung des Uran(VI) betrachtet. Mit Hilfe der zeitaufgelösten laserinduzierten Fluoreszenzspektroskopie bei tiefen Temperaturen wurde dabei ein Uranyl(VI)-Glucose-Komplex nachgewiesen. Die Komplexierung wurde lediglich bei pH 5 beobachtet und weist eine Komplexbildungskonstante von log βI=0,1 M = 15,25 ± 0,96 für den [UO2(C6H12O6)]2+-Komplex auf. Mit einer Fluoreszenzlebensdauer von 20,9 ± 2,9 µs und den Hauptemissionsbanden bei 499,0nm, 512,1 nm, 525,2 nm, 541,7 nm und 559,3 nm konnte der Uranyl(VI)-Glucose-Komplex fluoreszenzspektroskopisch charakterisiert werden. Unter reduzierenden Bedingungen wurde, ab pH-Wert 4 eine auftretende Umwandlung vom sechswertigen zum vierwertigen Uran durch Glucose in Gegenwart von Licht beobachtet. Der Anteil an gebildetem Uran(IV) steigt asymptotischen bis zu einem pH-Wert von 9, wo das Maximum mit 16 % bestimmt wurde. Als Reaktionsprodukt der Redoxreaktion wurde eine Uran(VI)-Uran(IV)-Mischphase mit der Summenformel [UIV(UVIO2)5(OH)2]12+ identifiziert. Mit Hilfe der cryo-TRLFS wurde, durch Verminderung von Quencheffekten die Uranspeziation in natürlichen Medien (Urin, Mineralwasser) direkt bestimmt. Proben mit Uran Konzentrationen von < 0,1 µg/L konnten dadurch analysiert werden. In handelsüblichen Mineralwässern wurde die zu erwartende Komplexierung durch Carbonat nachgewiesen. Im Urin zeigte sich in Abhängigkeit vom pH-Wert eine unterschiedliche Uranspeziation. Die fluoreszenzspektroskopische Untersuchung wies bei niedrigerem pH Wert (pH<6) eine Mischung aus Citrat- und Phosphat-Komplexierung des U(VI) und bei höheren pH-Wert (pH>6) eine deutliche Beteilung von Carbonat an der Komplexierung auf. Diese Ergebnisse stehen in sehr guter Übereinstimmung mit theoretischen Modellrechnungen zur Uranspeziation im Urin. Die in dieser Arbeit gewonnenen Ergebnisse zeigen, dass für eine zuverlässigere Prognose des Urantransportes in Geo- und Biosphäre in Zukunft nicht nur Betrachtungen zur Komplexchemie, sondern auch zum Redoxverhalten des Urans nötig sind, um die Mobilität in der Natur richtig abschätzen zu können.

The presence of ultraviolet patterns on body, as well as perception of ultraviolet spectrum by special photoreceptors, is part of sensory ecology of many animal species, including reptiles. Most current research discusses the importance of ultraviolet signs in coloration of diurnal species. The aim of our study was to find out what character have the reflective signs in ultraviolet spectrum in overall coloration of common leopard gecko (Eubplepharis macularius) through a digital photography. The reflective pattern is present in both adults and juveniles and passes as well as the rest of the coloration by significant ontogenetic changes. Another aim was to evaluate the role of ultraviolet reflecting signs in the biology of this crepuscular-nocturnal species. We expect that the pattern contributes to their antipredatory strategies during their first few months of life, and also a white reflecting surface is preserved in adulthood, especially on their tail, which is differently coloured than the rest of the body.