Title from PDF of title page (University of Missouri--Columbia, viewed on Feb 24, 2010). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Dissertation advisor: Dr. Gary Stacey. Vita. Includes bibliographical references.

Thesis (Ph.D.) - University of St Andrews, May 2008.

Thesis (Ph. D.)--Drexel University, 2007. / Includes abstract and vita. Includes bibliographical references (leaves 103-115).

Thesis (Ph.D.)--University of Delaware, 2007. / Principal faculty advisor: Jeremy S. Edwards, Dept. of Chemical Engineering. Includes bibliographical references.

No description available.

Philosophiae Doctor - PhD / In this thesis, we have firstly cloned and expressed the domains that harbours the putative catalytic GC domain in these receptor molecules and demonstrate that these molecules can convert GTP to cGMP in vitro. Secondly, we show that exogenous application of both Phytosulfokine and Brassinosteroid increase changes of intracellular cGMP levels in Arabidopsis mesophyll protoplast demonstrating that these molecules have GC activity in vivo and therefore provide a link as second messenger between the hormones and down-stream responses. In order to elucidate a relationship between the kinase and GC domains of the PSK receptor, we have used the AtPSKR1 receptor as a model and show that it has Serine/Threonine kinase activity using the Ser/Thr peptide 1 as a substrate. In addition, we show that the receptor`s ability to phosphorylate a substrate is affected by the product (cGMP) of its co-domain (GC) and that the receptor autophosphorylates on serine residues and this step was also observed to be affected by cGMP. When Arabidopsis plants are treated with a cell permeable analogue of cGMP, we note that this can affect changes in the phosphoproteome in Arabidopsis and conclude therefore that the cGMP plays a role in kinase-dependent downstream signalling. The obtained results suggest that the receptor molecules investigated here belong to a novel class of GCs that contains both a cytosolic kinase and GC domains, and thus have a domain organisation that is not dissimilar to that of atrial natriuretic peptide receptors NPR1 and NPR2. The findings also strongly suggest that cGMP has a role as a second messenger in both Brassinosteroid and Phytosulfokine signalling. We speculate that other proteins with similar domain organisations may also have dual catalytic activities and that a significant number of GCs, both in plants and animals, remain to be discovered and characterised. / South Africa

No abstract available / Dissertation (MSc (Bioinformatics))--University of Pretoria, 2007. / Biochemistry / MSc / unrestricted

Les chloroéthènes sont les polluants majeurs des eaux souterraines et des nappes phréatiques. De par leur toxicité et leur effet cancérigène, ils représentent une préoccupation majeure pour les autorités publiques et sanitaires. La restauration des sites contaminés est possible par des techniques de dépollution biologique impliquant les microorganismes (bioremédiation microbienne). Cependant, la réussite des traitements dépend à la fois des conditions physicochimiques du site pollué et des capacités de dégradation de la microflore indigène. Ainsi, pour optimiser les processus de décontamination, l’identification et le suivi des différentes populations microbiennes sont indispensables avant et pendant le traitement. Les biopuces ADN fonctionnelles (FGA, Functional Gene Array), outils moléculaires haut débit, sont particulièrement bien adaptées pour des applications en bioremédiation. Leur élaboration nécessite de disposer de logiciels performants pour le design de sondes qui combinent à la fois une forte sensibilité, une très bonne spécificité et un caractère exploratoire, ce dernier étant indispensable pour la détection des séquences connues mais surtout de celles encore jamais décrites au sein d’échantillons environnementaux. Un nouveau logiciel, autorisant la sélection de sondes combinant tous ces critères, a été développé et nommé HiSpOD. Son utilisation pour la construction d’une FGA dédiée aux voies de biodégradation des chloroéthènes a permis d’évaluer l’effet de traitements de biostimulation sur la microflore indigène pour plusieurs sites industriels contaminés. Les données révèlent différentes associations entre microorganismes déhalorespirants qui sont fonction des paramètres environnementaux. / Chlorinated solvents are among the most frequent contaminants found in groundwater and subsurface ecosystems. Because of their high toxicity and carcinogenicity, they represent a serious risk for human health and the environment. Thus, such polluted sites need a rehabilitation treatment. Among remediation solutions, microbial bioremediation represents a less invasive and expensive alternative than physico-chemical treatments. However, the process efficiency greatly depends on the environmental conditions and the microbial populations’ biodegradation capacities. Therefore, bioremediation treatment optimization requires the identification and monitoring of such capacities before and during the treatment. Functional Gene Arrays (FGA), by profiling environmental communities in a flexible and easy-to-use manner, are well adapted for an application in bioremediation. But, constructing efficient microarrays dedicated to microbial ecology requires a probe design step allowing the selection of highly sensitive, specific and explorative oligonucleotides. After a detailed state of the art on probe design strategies suitable for microbial ecology studies, we present new software, called HiSpOD, generating efficient explorative probes for FGA dedicated to environmental applications. Finally, this bioinformatics tool was used to construct a FGA targeting most genes involved in chloroethenes biodegradation pathways which allowed the evaluation of biostimulation treatments conducted on indigenous bacterial populations for several industrial contaminated sites.

Toda medida, desde que feita por um instrumento real, tem uma imprecisão associada. Neste trabalho, abordamos a questão das imprecisões em experimentos de microarranjos de cDNA de dois canais, uma tecnologia que tem sido muito explorada nos últimos anos e que ainda é um importante auxiliar nos estudos de expressões gênicas. Dezenas de milhares de representantes de genes são impressos em uma lâmina de vidro e hibridizados simultaneamente com RNA mensageiro de duas amostras diferentes de células. Essas amostras são marcadas com corantes fluorescentes diferentes e a lâmina, após a hibridização, é digitalizada, obtendo-se duas imagens. As imagens são analisadas com programas especiais que segmentam os locais que estavam os genes e extraem estatísticas dos píxeis de cada local. Por exemplo, a média, a mediana e a variância das intensidades do conjunto de píxeis de cada local (o mesmo é feito normalmente para uma área em volta de cada local, chamada de fundo). Estimadores estatísticos como o da variância nos dão uma estimativa de quão precisa é uma certa medida. Uma vez de posse das estimativas das intensidades de cada local, para se obter a efetiva expressão de um gene, algumas transformações são feitas nos dados de forma a eliminar variabilidades sistemáticas. Neste trabalho, mostramos como podem ser feitas as análises a partir de uma medida de expressão gênica com um erro estimado. Mostramos como estimar essa imprecisão e estudamos, em termos de propagação da imprecisão, os efeitos de algumas transformações realizadas nos dados, por exemplo, a remoção do viés estimado pelo método de regressão local robusta, mais conhecido como \\textit{lowess}. Uma vez obtidas as estimativas das imprecisões propagadas, mostramos também como utilizá-las na determinação dos genes diferencialmente expressos entre as amostras estudadas. Por fim, comparamos os resultados com os obtidos por formas clássicas de análise, em que são desconsideradas as imprecisões das medidas. Concluímos que a modelagem das imprecisões das medidas pode favorecer as análises, já que os resultados obtidos em uma aplicação com dados reais de expressões gênicas foram condizentes com os que encontramos na literatura. / Any measurement, since it is made for a real instrument, has an uncertainty associated with it. In the present paper, we address this issue of uncertainty in two-channel cDNA Microarray experiments, a technology that has been widely used in recent years and is still an important tool for gene expression studies. Tens of thousands of gene representatives are printed onto a glass slide and hybridized simultaneously with mRNA from two different cell samples. Different fluorescent dyes are used for labeling both samples. After hybridization, the glass slide is scanned yielding two images. Image processing and analysis programs are used for spot segmentation and pixel statistics computation, for instance, the mean, median and variance of pixel intensities for each spot. The same statistics are computed for the pixel intensities in the background region. Statistical estimators such as the variance gives us an estimate of the accuracy of a measurement. Based on the intensity estimates for each spot, some data transformations are applied in order to eliminate systematic variability so we can obtain the effective gene expression. This paper shows how to analyze gene expression measurements with an estimated error. We presented an estimate of this uncertainty and we studied, in terms of error propagation, the effects of some data transformations. An example of data transformation is the correction of the bias estimated by a robust local regression method, also known as \\textit{lowess}. With the propagated errors obtained, we also showed how to use them for detecting differentially expressed genes between different conditions. Finally, we compared the results with those obtained by classical analysis methods, in which the measurement errors are disregarded. We conclude that modeling the measurements uncertainties can improve the analysis, since the results obtained in a real gene expressions data base were consistent with the literature.

No description available.