Tuberculosis, caused by the intracellular pathogen Mycobacterium tuberculosis, is one of the most prevalent infectious diseases in our world today. In order to survive within the host the bacteria need to sense and respond to changes in the environment; however, signal transduction in this bacterium is poorly understood. PknB is a serine/threonine kinase essential for the in vitro survival of M. tuberculosis and therefore a potential drug target against the bacteria. It is the goal of the current study to elucidate downstream substrates of PknB. We have found that PknB shares in vitro substrates with another serine/threonine kinase, PknH, implying the potential complexity of the signaling pathways in the bacteria. We have also provided the first description of the coupling between serine/threonine kinases PknB and PknH with a two-component system response regulator DevR, and further proposed Ser/Thr phosphorylation as the negative regulator of DevR transcription activator activity based on LC-MS/MS analysis. Finally, we have identified a previously unknown phosphoprotein glyceraldehyde 3-phosphate dehydrogenase encoded by the ORF Rv1436, which demonstrates autophosphorylation activity and which phosphorylation is independent of PknB. Overall, the current study has contributed to advance our understanding of the signal transduction pathways and phosphoproteome in Mycobacterium tuberculosis.

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Asymmetric hydrophosphination, the addition of a P-H bond across a C-C double bond, is an attractive potential route to chiral phosphines, which have important applications in many other types of asymmetric catalysis. However, a highly active and stereoselective catalyst for this reaction has yet to be identified. The ruthenium indenyl complex [RuCl(η5-indenyl)(PPh3)2] (1) was investigated as a potential catalyst for hydrophosphination through an exploration of the steps involved in this process: substrate coordination, P-H bond activation, and P-C bond formation. Substitution of triphenylphosphine ligands at the metal centre of 1 by alkyl- and aryl-substituted secondary phosphines (PR2H: R = Cy (a), Pri (b), Et (c), Ph (d) or Tolp (e)) gave predominantly the monosubstituted secondary phosphine complexes [RuCl(η5-indenyl)(PR2H)(PPh3)] (3a-e). Hydride ([RuH(η5-indenyl)(PR2H)(PPh3)] (6a,d)) and cationic nitrile ([Ru(η5-indenyl)(NCR')(PR2H)(PPh3)][PF6] (7a,d: R' = CH=CH2; 8a-b,d: R = CH3)) derivatives of 3 were prepared and in all cases the potentially reactive P-H bond of the secondary phosphine ligand did not interfere with the chemical transformation. Deprotonation of the P-H bond of the bulky dialkylphosphine-substituted chloro complexes 3a-b with KOBut gave five-coordinate, planar terminal phosphido complexes [Ru(η5-indenyl)(PR2)(PPh3)] (10a-b) that contain a unique Ru-PR2 π-bond. The analogous phosphido complexes 10d-e, containing less bulky aryl substituents at phosphorus, were found to be unstable at room temperature and were observed only by low temperature 31P{1H} NMR spectroscopy. Phosphido complexes 10a-b were found to be highly P-basic, capable of deprotonating the C-H bond of acetonitrile (pKa = 24) to give the metallated acetonitrile complex [Ru(CH2CN)(η5-indenyl)(PR2H)(PPh3)] (9a-b), and to be very P-nucleophilic, reacting with iodomethane (MeI) to give a new P-C bond in [RuI(η5-indenyl)(PCy2Me)(PPh3)] (17a). As might be expected, the addition of donor ligands to low-coordinate 10a-b was found to disrupt the Ru-PR2 π-bond to give six-coordinate terminal phosphido complexes [Ru(η5-indenyl)(L)(PR2H)(PPh3)], with pyramidal, instead of planar, geometry at phosphorus. These additions are irreversible in the case of CO (19a-b) or PCy2H (21a), while pyridine (23a-b) or NCPh (24a-b) adducts were shown by 31P{1H} NMR spectroscopy to be in equilibrium with 10a-b and the uncoordinated ligand. The addition of known substrates for transition metal-mediated hydrophosphination, phenylacetylene and acrylonitrile, to 10a-b resulted in a [2+2] cycloaddition of the unsaturated C-C bond at the Ru-PR2 π-bond to give metallacyclic complexes [Ru(η5-indenyl)(κ2-PhC=CHPR2)(PPh3)] (27a-b) and [Ru(η5-indenyl)(κ2-NCCHCH2PR2)(PPh3)] (32a-b) respectively. Surprisingly the addition of simple non-activated olefins (i.e. ethylene, 1-hexene, or norbornene), which were not previously known to be active substrates for this reaction, also gave [2+2] cycloaddition products. These cycloaddition reactions were found to be 100% regioselective, and are also stereoselective in the case of substituted alkenes (>96%). Experimental evidence suggests that these P-C bond forming reactions proceed via a concerted [2+2] cycloaddition pathway.

The preparation and characterisation of chelating sorbents suitable for the high efficiency separation of trace metals in complex samples, using a single column and isocratic elution, is described. Hydrophobic, neutral polystyrene divinylbenzene resins were either impregnated with chelating dyes or dynamically modified with heterocyclic organic acids, using physical adsorption and chemisorption processes respectively. A hydrophilic silica substrate was covalently bonded with a chelating aminomethylphosphonic acid group, to assess the chelating potential of this molecule. These substrates were characterised in terms of metal retention capability (selectivity coefficients and capacity factors), separation performance, column efficiency and suitability for analytical applications. Chelating molecules with different ligand groups were found to have unique selectivity patterns dependant upon the conditional stability constants of the chelate. Other factors, including mobile phase constituents - complexing agents, ionic strength and pH, column length and column capacity were additionally investigated to examine their effect upon the separation profiles achieved. The promising metal separation abilities illustrated by a number of these chelating columns were exploited for the determination of trace toxic metals in complex sample matrices using High Performance Chelation Ion Chromatography (HPCIC). This included the determination of beryllium in a certified stream sediment, uranium in seawater and a certified stream sediment, and cadmium, lead and copper in a certified rice flour. The results for each analysis fell within the certified limits, and reproducibility was good. The optimisation of post column detection systems using chromogenic ligands additionally gave good detection limits for the metals in each separation system.

With the industry striving for smaller devices, new technologies are developed to further miniaturize electronics devices. To this end, realization of 3D/non-planar interconnects, which aim at miniaturizing the interconnects formed between components on the same device, has attracted many researchers. This thesis focuses on a feasibility analysis for developing non-planar interconnects on various flexible substrates using laser assisted maskless microdeposition (LAMM), which is a pressure-less process. There are two types of flexible substrates that are used: double-sided copper substrates separated by a layer of polyethylene terephthalate (PET) as well as a polyethylene terephthalate flexible substrate with surface-mounted resistors. For both substrates, multiple types of experiments were conducted to discover procedures which result in the highest rate of success for forming conductive interconnects. Optimal process parameters and deposition techniques were determined after multiple experiments. After experiments were completed, the resultant substrates were subject to various characterization methodologies including optical and scanning electron microscopy, energy-dispersive X-ray spectroscopy, X-ray diffraction and profilometery. The results of these methodologies are documented in this thesis. After many types of experiments involving substrate manipulation of the double-sided copper substrates, it was shown that the silver nano-particles were more likely to form a conductive interconnect when a polished slant was fabricated on the substrate. Many deposition patterns were used for the flexible substrates with surface-mounted resistors. Of these patterns, the two patterns, the ‘zigzag’ and ‘dot solder’ patterns, proved to have a much higher success rate for creating conductive interconnects compared to the other patterns. During this study, the results of the experiments using the LAMM process show that this technology has great potential for creating non-planar interconnects on flexible substrates. The experiments however suggest that the process is very sensitive to the material composition and process parameters. As such, with a small change in parameters, the 3D interconnects can fail to be produced. It was also observed that the possibility of silver interconnect fractures is higher where dissimilar materials with different thermal expansion rates are used for the underlying substrates.

Tuberculosis, caused by the intracellular pathogen Mycobacterium tuberculosis, is one of the most prevalent infectious diseases in our world today. In order to survive within the host the bacteria need to sense and respond to changes in the environment; however, signal transduction in this bacterium is poorly understood. PknB is a serine/threonine kinase essential for the in vitro survival of M. tuberculosis and therefore a potential drug target against the bacteria. It is the goal of the current study to elucidate downstream substrates of PknB. We have found that PknB shares in vitro substrates with another serine/threonine kinase, PknH, implying the potential complexity of the signaling pathways in the bacteria. We have also provided the first description of the coupling between serine/threonine kinases PknB and PknH with a two-component system response regulator DevR, and further proposed Ser/Thr phosphorylation as the negative regulator of DevR transcription activator activity based on LC-MS/MS analysis. Finally, we have identified a previously unknown phosphoprotein glyceraldehyde 3-phosphate dehydrogenase encoded by the ORF Rv1436, which demonstrates autophosphorylation activity and which phosphorylation is independent of PknB. Overall, the current study has contributed to advance our understanding of the signal transduction pathways and phosphoproteome in Mycobacterium tuberculosis.

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abstract: HgCdTe is currently the dominant material for infrared sensing and imaging, and is usually grown on lattice-matched bulk CdZnTe (CZT) substrates. There have been significant recent efforts to identify alternative substrates to CZT as well as alternative detector materials to HgCdTe. In this dissertation research, a wide range of transmission electron microscopy (TEM) imaging and analytical techniques was used in the characterization of epitaxial HgCdTe and related materials and substrates for third generation IR detectors. ZnTe layers grown on Si substrates are considered to be promising candidates for lattice-matched, large-area, and low-cost composite substrates for deposition of II-VI and III-V compound semiconductors with lattice constants near 6.1 Å. After optimizing MBE growth conditions including substrate pretreatment prior to film growth, as well as nucleation and growth temperatures, thick ZnTe/Si films with high crystallinity, low defect density, and excellent surface morphology were achieved. Changes in the Zn/Te flux ratio used during growth were also investigated. Small-probe microanalysis confirmed that a small amount of As was present at the ZnTe/Si interface. A microstructural study of HgCdTe/CdTe/GaAs (211)B and CdTe/GaAs (211)B heterostructures grown using MBE was carried out. High quality MBE-grown CdTe on GaAs(211)B substrates was demonstrated to be a viable composite substrate platform for HgCdTe growth. In addition, analysis of interfacial misfit dislocations and residual strain showed that the CdTe/GaAs interface was fully relaxed. In the case of HgCdTe/CdTe/ GaAs(211)B, thin HgTe buffer layers between HgCdTe and CdTe were also investigated for improving the HgCdTe crystal quality. A set of ZnTe layers epitaxially grown on GaSb(211)B substrates using MBE was studied using high resolution X-ray diffraction (HRXRD) measurements and TEM characterization in order to investigate conditions for defect-free growth. HRXRD results gave critical thickness estimates between 350 nm and 375 nm, in good agreement with theoretical predictions. Moreover, TEM results confirmed that ZnTe layers with thicknesses of 350 nm had highly coherent interfaces and very low dislocation densities, unlike samples with the thicker ZnTe layers. / Dissertation/Thesis / Ph.D. Materials Science and Engineering 2012

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Tanto no meio rural como urbano as novas tendências são de buscar uma melhoria de técnicas e de uso de materiais alternativos buscando-se uma maior sustentabilidade. A utilização de plantas nas paredes e telhados é um dos mais recentes campos da pesquisa ambiental e busca encontrar uma solução ecológica e sustentável para melhorar a qualidade de vida urbana e rural. Uma cobertura verde consiste de um substrato leve e de uma vegetação apropriada, plantada sobre uma base impermeável. Podem conter também camadas adicionais, tais como, um sistema de drenagem e irrigação e uma barreira anti-raízes. Os efeitos positivos da vegetação sobre o ambiente urbano já são bem conhecidos e neste caso da cobertura, diminuem as enxurradas, filtro biológico, redução da poluição do ar, amenizam o calor nas edificações durante o verão e o conservam durante o inverno. Há também benefícios para a fauna, com retorno de espécies que mantêm o equilíbrio biológico local. Essas coberturas podem ter muitas aplicações, como em indústrias, residências, escritórios e outras propriedades comerciais e rurais. Uma criteriosa seleção de plantas permite que a cobertura verde tenha sucesso em condições adversas. Por outro lado, são necessárias pesquisas no sentido de assegurar um meio de cultura que retenha água e ao mesmo tempo proporcione uma boa drenagem para que as raízes não apodreçam, permitindo uma maior gama de espécies de plantas na cobertura. Neste sentido, realizou-se um estudo dos substratos mais abundantes na região, buscando tecnologias apropriadas para a confecção dos telhados verdes e identificando as melhores técnicas de aplicação, de acordo com a necessidade de cada ambiente. Para a realização deste trabalho foram utilizados dez tipos de substratos que foram pré-selecionados de acordo com a disponibilidade no mercado, de mais fácil obtenção e de melhor custo beneficio ambiental. As análises estatísticas utilizam o delineamento experimental inteiramente ao acaso, em esquema fatorial com três repetições e utilizando o programa estatístico SAS para auxiliar na análise e confecção dos gráficos e discussões. Na avaliação do conjunto de características, a turfa marrom (TM) apresentou o menor valor de densidade seca, maior porosidade total, maiores valores de água facilmente disponível, água disponível e capacidade de retenção de água, e mesmo valor de espaço de aeração daquele sugerido como referência sendo um bom material para o uso em telhados verdes, formando compostos com outros materiais e buscando as melhores características e sustentabilidade do sistema. Dentre os substratos comerciais estudados, o substrato para espécies hortícolas (SH) apresentou os maiores valores de capacidade de retenção de água e água disponível para as plantas, possibilitando um maior intervalo entre irrigações ou chuvas, características importantes para o uso em telhados verdes. No entanto, tornam-se necessários mais estudos com a finalidade identificar mais materiais e substratos apropriados para o uso em telhados verdes, levando-se em conta as diferentes características de cada local, tipo de vegetação escolhida, capacidade de suporte da edificação, dentre outro fatores. / Rural and urban new trends seek an improvement in techniques and use of alternative materials for greater sustainability. The use of plants on walls and roofs is one of the newest fields of environmental research, looking for a green and sustainable solution to improve the quality of urban and rural life. A green cover consists of a substrate and appropriate vegetation planted on an impermeable base. It can also contain additional layers, such as a system of drainage and irrigation with an anti-root barrier. The positive effects of roof vegetation on urban environment are well known, as reducing the runoff,acting as a biological filter, reducing air pollution, minimizing heat inside the buildings during summer and maximizing during the winter. There are also benefits to fauna, with the return of various species, maintaining the local biological balance. The coverage can be adapted to industries, homes, offices and other commercial and rural properties. Careful selection of plants allows the green roof to succeed in adverse conditions. In addition, research is necessary to ensure a culture medium that retains water that the same time provides good drainage to prevent root rot, allowing a wider range of plant species in the cover. In this sense, a study was conducted involving the most abundant substrates of our region, seeking appropriate technologies for the manufacture of green roofs and identifying the best application techniques, according to the needs of each environment. For this work ten types of preselected substrates were used according to market availability, easier to obtain and with a better environmental cost/benefit ratio. Statistics of the experimental design was completely randomized in a factorial format with three replications and using the SAS statistical software to assist in the analysis and construction of graphs and discussions. In evaluating the feature of the cover set, brown peat (TM) presented the lowest value of dry density, higher porosity, higher values of easily available water, available water capacity and water retention, and even the amount of aeration space, suggested TM as being a good reference material for use in green roofs. Among the studied commercial substrates, the substrate for horticultural species (SH) showed the highest values of water holding capacity and water available to plants, allowing a longer interval between irrigations or rainfall, an important characteristic for its use on green roofs. However, more studies are needed in order to identify the most suitable materials and for use in green roof substrates, taking into account the different characteristics of each site, vegetation type, bearing capacity of the building, among other factors.