Coupled two-dimensional HPLC systems were developed for the separation of complex sample matrices. Low molecular weight polystyrene oligomers were used as model compounds for the development of these systems since the sample dimensionality of oligostyrenes could be defined and classified according to two sample characteristics. That is, oligostyrenes could be classed as a two-dimensional sample, where one sample dimension is the variation in the number of monomers that make up oligomeric units determining the weight of the polymer, with a second sample dimension being the variation in stereoisomerism of each oligomer. During this study it was found that by combining two separation dimensions consisting of C18 and carbon clad zirconia (CCZ) phases separation of oligostyrenes according to molecular weight and stereochemistry was possible in coupled two-dimensional systems. The C18 phase separated the sample according to molecular weight, while the CCZ phase separated the sample according to the stereochemistry. The stereoisomer separations of the oligostyrenes reported on the CCZ surface were far superior to those previously reported in the literature. The efficiency of the separation process in a coupled two-dimensional system was then evaluated by studying the variation in band variance of a test probe. The results of the study indicated that the performance of a two-dimensional separation process was highly dependent upon the mobile phase compatibility, even when solvents were completely miscible and of similar polarities. Extracts from 17 species of Australian native plants were examined for xanthine oxidase inhibitory activity, the enzyme responsible for the formation of the disease gout. Chromatographic separation was conducted on plant extracts found to possess significant inhibitory activity against xanthine oxidase, with an extract from the species Clerodendrum floribundum R. Br. found to possess the greatest activity of the species examined. A two-dimensional separation of a crude extract from Clerodendrum floribundum R. Br. was conducted using one of the developed HPLC systems, to illustrate the use of such a system for the separation of a non-model complex sample mixture / Doctor of Philosophy (PhD)

<p>G protein-coupled receptors (GPCR) are integral membrane proteins with seven α-helices that translate a remarkable diversity of signals into cellular responses. The superfamily of GPCRs is among the largest and most diverse protein families in vertebrates. </p><p>We have searched the human genome for GPCRs and show that the family includes approximately 800 proteins, which can divided into five main families; <i>Glutamate</i>, <i>Rhodopsin</i>, <i>Adhesion</i>, <i>Frizzled/Taste2</i> and <i>Secretin</i>. This study represents one of the first overall road maps of the GPCR family in a mammalian genome. Moreover, we identified eight novel members of the human <i>Adhesion</i> family which are characterized by long N-termini with various domains. We also investigated the GPCR repertoire of the chicken genome, where we manually verified a total of 557 chicken GPCRs. We detected several specific expansions and deletions that may reflect some of the functional differences between human and chicken.</p><p>Substantial effort has been made over the years to find compounds that can bind and activate the melanocortin 4 receptor (MC4R). This receptor is involved in food intake and is thus an important target for antiobesity drugs. We used site-directed mutagenesis to insert micromolar affinity binding sites for zinc between transmembrane (TM) regions 2 and 3. We generated a molecular model of the human MC4R which suggests that a rotation of TM3 is important for activation of the MC4R. </p><p>Furthermore, we present seven new vertebrate prolactin releasing hormone receptors (PRLHRs) and show that they form two separate subtypes, PRLHR1 and PRLHR2. We performed a pharmacological characterization of the human PRLHR which showed that the receptor can bind neuropeptide Y (NPY) related ligands. We propose that an ancestral PRLH peptide has coevolved with a redundant NPY binding receptor, which then became PRLHR. This suggests how gene duplication events can lead to novel peptide ligand/receptor interactions and hence spur the evolution of new physiological functions. </p>

<p>G protein-coupled receptors (GPCRs) are signal mediators that have a prominent role in the regulation of physiological processes and they make up the targets for 30-45% of all drugs. </p><p>Papers I and II describe the discovery of new human GPCRs belonging to the Rhodopsin family, a family which contains many common drug targets. The new receptors have only weak relationships to previously known GPCRs. However, they have been evolutionary conserved in several species and most of them display distinct expression patterns.</p><p>In paper III we identified new human GPCRs belonging to the Adhesion family, which is characterised by very long N-termini containing conserved domains. The different compositions of conserved domains as well as the expression patterns suggest that the Adhesions can have several different functions.</p><p>In paper IV we revealed remarkable species variations in the repertoires of Trace Amine-Associated Receptors (TAARs), which are relatives of the biogenic amine receptors. The human, mouse and rat TAAR genes are located in only one locus and are therefore most likely the result of gene tandem duplications. 47 of the 57 zebrafish TAARs were mapped to nine different loci on six chromosomes containing from 1 to 27 genes each. This study suggests that the TAARs arose through several different mechanisms involving tetraploidisation, block duplications, and local duplication events. </p><p>Papers V and VI are overall analyses of the repertoires of GPCRs in humans, mice and rats; which contain approximately 800, 1800 and 1900 members, respectively. The repertoires were compared to distinguish between species-specific and common (orthologous) members, something which is important for example when predicting drug effects from experiments in rodents. The Glutamate, Adhesion, Frizzled and Secretin families show no or very little variation between human and rodents, whereas the repertoires of olfactory, vomeronasal and Taste2 receptors display large differences between all three species. </p>

<p>A new approach to analysis of protein-ligand interactions, termed proteochemometrics, has been developed. Contrary to traditional quantitative structure-activity relationship (QSAR) methods that aim to correlate a description of ligands to their interactions with one particular target protein, proteochemometrics considers many targets simultaneously.</p><p>Proteochemometrics thus analyzes the experimentally determined protein-ligand interaction activity data by correlating the data to a complex description of all interaction partners and; in a more general case even to interaction environment and assaying conditions, as well. In this way, a proteochemometric model analyzes an “interaction space,” from which only one cross-section would be contemplated by any one QSAR model.</p><p>Proteochemometric models reveal the physicochemical and structural properties that are essential for protein-ligand complementarity and determine specificity of molecular interactions. From a drug design perspective, models may find use in the design of drugs with improved selectivity and in the design of drugs for multiple targets, such as mutated proteins (e.g., drug resistant mutations of pathogens).</p><p>In this thesis, a general concept for creating of proteochemometric models and approaches for validation and interpretation of models are presented. Different types of physicochemical and structural description of ligands and macromolecules are evaluated; mathematical algorithms for proteochemometric modeling, in particular for analysis of large-scale data sets, are developed. Artificial chimeric proteins constructed according to principles of statistical design are used to derive high-resolution models for small classes of proteins.</p><p>The studies of this thesis use data sets comprising ligand interactions with several families of G protein-coupled receptors. The presented approach is, however, general and can be applied to study molecular recognition mechanisms of any class of drug targets.</p>

Glass forming ability (GFA) in the Pr-rich Pr-(Cu, Ni)-Al alloys at or near the eutectic points was systematically studied. It was found that the GFA in the pseudo-ternary alloys of Pr-(Cu, Ni)-Al is higher than that of the ternary alloys of Pr-Cu-Al. Two eutectic compositions in Pr-(Cu, Ni)-Al alloys were found by DSC, namely, Pr₆₈(Cu₀.₅Ni₀.₅)₂₅Al₇ and Pr₅₂(Cu₀.₅Ni₀.₅)₂₅Al₂₃ (at %). The later one shows better GFA than the first one. However, the best GFA was obtained at an off-eutectic composition of Pr₅₄(Cu₀.₅Ni₀.₅)₃₀Al₁₆, which can be formed in fully amorphous rod with diameter of 1.5 mm by copper mould casting. The deviation of the best GFA composition from the eutectic point [Pr₆₈(Cu₀.₅Ni₀.₅)₂₅Al₇] was explained in terms of the asymmetric coupled eutectic zone and the higher glass transition temperature Tg on the hypereutectic side. / Singapore-MIT Alliance (SMA)

Coalbeds are an extremely complicated porous medium with characteristics of heterogeneity, dual porosity and stress sensitivity. In the past decades great achievements have been made to the simulation models of pressure depletion coalbed methane (CBM) recovery process and CO2 sequestration and enhanced coalbed methane (ECBM) recovery process. However, some important mechanisms are still not or not properly included. Among them, the influence of geomechanics is probably the most important one. Because of its influence coalbed permeability, the key parameter for the success of recovery processes, changes drastically with alterations of in situ stresses and strains during these processes. In present reservoir simulators, the change of coalbed permeability is estimated with analytical models. Due to the assumptions and over simplifications analytical models have limitations or problems in application. In this research to properly estimate the changes of permeability and porosity in the simulation of CO2 sequestration and ECBM recovery process, comprehensive permeability and porosity models have been developed with minimum assumptions and simulation methods established. Firstly, a set of continuum medium porosity and permeability coupling models is built up and a simulation procedure to apply these models in reservoir and geomechanical coupled simulations proposed. Using the models and simulation procedure a sensitivity study, mainly on the parameters related to coalbed permeability change and deformation, has been made for the CBM recovery process. Then based on the understanding, a set of discontinuum medium porosity and permeability coupling models is developed and a procedure to apply these models in reservoir and geomechanical coupled simulations presented. The new models are more comprehensive and adaptable, and can accommodate a wide range of coalbeds and in situ conditions. The proposed equivalent continuum deformation model for coal mass is validated by simulating a set of lab tests including a uniaxial compression test in vacuum and a CO2 swelling test under axial constraint in the longitudinal (vertical) direction. At last the discontinuum medium porosity and permeability coupling models and the simulation procedure are successfully applied to simulate part of a series of micro-pilot tests of ECBM and CO2 sequestration at Fenn Big Valley of Alberta, Canada. / Geotechnical Eengineering

A multilevel Space Vector PWM (SVPWM) technique is developed for a 3-level 3-phase PWM Voltage Source Inverter using a 3-phase coupled inductor to ensure high performance operation. The selection of a suitable PWM switching scheme for the Coupled Inductor Inverter (CII) topology should be based on the dual requirements for a high-quality multilevel PWM output voltage together with the need to minimize high frequency currents and associated losses in the coupled inductor and the inverter switches. Compared to carrier-based multilevel PWM schemes, the space vector techniques provide a wider variety of choices of the available switching states and sequences. The precise identification of pulse placements in the SVPWM method is used to improve the CII performance. The successful operation of the CII topology over the full modulation range relies on selecting switching states where the coupled inductor presents a low winding current ripple and a high effective inductance between the upper and lower switches in each inverter leg. In addition to these requirements, the CII operation is affected by the imbalance inductor common mode dc current. When used efficiently, SVPWM allows for an appropriate balance between the need to properly manage the inductor winding currents and to achieve harmonic performance gains. A number of SVPWM strategies are developed, and suitable switching states are selected for these methods. Employing the interleaved PWM technique by using overlapping switching states, the interleaved Discontinuous SVPWM (DSVPWM) method, compared to other proposed SVPWM methods, doubles the effective switching frequency of the inverter outputs and, as a result, offers superior performance for the CII topology by reducing the inductor losses and switching losses. The inverter operation is examined by means of simulation and experimental testing. The experimental performance comparison is obtained for different PWM switching patterns. The inverter performance is affected by high-frequency inductor current ripple; the excessive inductor losses are reduced by the DSVPWM method. Additional experimental test results are carried out to obtain the inverter performance as a variable frequency drive when operated in steady-state and during transient conditions. The CII topology is shown to have great potential for variable speed drives. / Power Engineering and Power Electronics

For coupled systems of differential equations on networks, a graph-theoretic approach to the construction of Lyapunov functions is systematically developed in this thesis. Kirchhoffs Matrix-Tree Theorem in graph theory plays an essential role in the approachs development. The approach is successfully applied to several coupled systems well-known in the literature to demonstrate its applicability and effectiveness. / Applied Mathematics

A systematic procedure is designed to evaluate the role of stochastic forcing (SF) in El Nino ? Southern Oscillation (ENSO) exhibited by coupled ocean-atmosphere general circulation models (CGCMs). The procedure is applied on a 163-year run of a CGCM which has realistic representation of ENSO and SF. The realism of ENSO in the CGCM is determined by statistical comparison with a 29-year global reanalysis product. SF is extracted from both the CGCM and reanalysis as residual atmospheric variability uncoupled to the ocean. Further, the Madden-Julian Oscillation (MJO) and non-MJO components are isolated from SF. The CGCM stochastic components are compared to those from the reanalysis to validate their representation. A coupled ocean-atmosphere model of intermediate complexity is first forced with stochastic components from the reanalysis. The resulting ENSO is examined for realism to evaluate the strengths and weaknesses of the intermediate coupled model, which is then forced with the stochastic components from the CGCM. Results are diagnosed to investigate the role of SF. It is found that the SF can play an important role in ENSO in the CGCM, especially in its warm events. The role is similar to reanalysis SF in generating ENSO period and spring predictability barrier. However, unlike in case of the reanalysis, the seasonal dependence of ENSO variance in the CGCM does not seem to be originating from its SF. The contribution to statistics appears to be higher from the MJO component of SF compared to the non-MJO component. The intermediate model simulations also suggest that both in CGCM and nature, the SF operates on a weakly stable coupled system to produce ENSO variability.

Nonlinear dynamical systems theory has inspired a new set of useful tools to be applied in climate studies. In this work we presented specific examples where information has been gained by the application of methods from nonlinear dynamical systems theory. The main goal is to understand the relative importance of stochastic forcing versus deterministic coupling within the context of Coupled General Circulation Models. This work address this important subject by approaching this goal through the development of a hierarchy of models with increasing complexity that we assert contain the essential dynamics of ENSO. We examined the effect of noise in a low order model and found that it is not restricted to blurring the attractor trajectories in phase space, but includes important changes in the dynamics of the system. The main results indicate that the presence of noise in a nonlinear system has two different effects. The presence of noise acts to increase the maximum Lyapunov exponent and can result in noise induced chaos if the system was originally stable. However, the same arguments are not valid if the original system is already in the chaotic regime, where the noise inclusion acts to decrease the maximum Lyapunov exponent, therefore increasing the system stability. The system of interest includes coupled ocean-atmosphere interactions and here we mimic this interaction by coupling two low order models with very different dominant time scales. These subsystems interact in a complex, nonlinear way and the behavior of the whole system cannot be explained by a linear summation of dynamics of the system parts. We used information theory concepts to detect the influence of the slow system dynamics in synchronizing the fast system in coupled models. We introduced a fast-slow coupled system, where both the slowness of the ocean model and the intensity of the boundary forcing anomalies contribute to the asymmetry and phase locking of both subsystems. The mechanisms controlling the fast modelspread were uncovered revealing uncertainty dynamics depending on the location of ensemble members in the model’s phase space. As an intermediate step between low order models and CGCMs we study the effect of noise on an intermediate complexity model. The addition of gaussian noise to the Zebiak-Cane model in order to understand the effects of noise on its attractor led to a way of estimating the noise level based on the effects of noise on the correlation dimension curves. We investigate the intrinsic predictability of the coupled models used here, and the different time scales associated with fast and slow modes were detected using the Finite Size Lyapunov Exponents. We found new estimates for the prediction horizon of ENSO for the Zebiak-Cane model as well as for the NCAR CCSM3 model and observations. The whole analysis of observations and CCSM3 was possible after applying noise reduction techniques. We also improved our understanding of three different noise reduction techniques by comparing the Local Projective Noise Reduction, the Interactive Ensemble strategy, and a Random Interactive Ensemble applied to CCSM3. The main difference between these two noise reduction techniques is when the process is applied. The Local Projective Noise Reduction can be applied to both model and observations, and it is done a posteriori in phase space, therefore the trajectories to be adjusted already posses the physical mechanisms embedded in them. The Interactive Ensemble approach can only be applied to model simulations and has shown to be a very useful technique for noise reduction since its done a priori while the system evolves instead of a posteriori, besides the fact that it allows to retrieve the spatial distribution of the noise level in physical space.