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A major driving force for development of new catalyst systems is the need for more efficient synthesis of chemical compounds essential to modern life. Catalysts having superior performance offer significant environmental and economic advantages, but their discovery is not trivial. Well-defined, homogeneous catalysts can offer unparalleled understanding of ligand effects, which proves invaluable in directing redesign strategies. This thesis work focuses on the design of ruthenium complexes for applications in dinitrogen activation and olefin metathesis. The complexes developed create new directions in small-molecule activation and asymmetric catalysis by late-metal complexes. Also examined are the dual challenges, ubiquitous in catalysis, of adequate interrogation of catalyst structure and performance. Insight into both is essential to enable correlation of ligand properties with catalyst activity and/or selectivity. Improved methods for accelerated assessment of catalyst performance are described, which expand high-throughput catalyst screening to encompass parallel acquisition of kinetic data. A final aspect focuses on direct examination of metal complexes, both as isolated species, and under catalytic conditions. Applications of charge-transfer MALDI mass spectrometry to structural elucidation in organometallic chemistry is described, and the technique is employed to gain insight into catalyst decomposition pathways under operating conditions.

The human body is made of billions of cells and nearly all have the same genome. However, there is a high diversity of cells, resulted from what part of the genome the cells use, i.e. which RNA molecules are expressed. Rapid advances within the field of sequencing allow us to determine the RNA molecules expressed in a specific cell at a certain time. The use of the new technologies has expanded our view of the human transcriptome and increased our understanding of when, where, and how each RNA molecule is expressed. The work presented in this thesis focuses on analysis of the human transcriptome. In Paper I, we describe an automated approach for sample preparation. This protocol was compared with the standard manual protocol, and we demonstrated that the automated version outperformed the manual process in terms of sample throughput while maintaining high reproducibility. Paper II addresses the impact of nuclear transcripts on gene expression. We compared total RNA from whole cells and from cytoplasm, showing that transcripts with long, structured 3’- and 5’-untranslated regions and transcripts with long protein coding sequences tended to be retained in the nucleus. This resulted in increased complexity of the total RNA fraction and fewer reads per unique transcript. Papers III and IV describe dynamics of the human muscle transcriptome. For Paper III, we systematically investigated the transcriptome and found remarkably high tissue homogeneity, however a large number of genes and isoforms were differentially expressed between genders. Paper IV describes transcriptome differences in response to repeated training. No transcriptome-based memory was observed, however a large number of isoforms and genes were affected by training. Paper V describes a transcript profiling protocol based on the method Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification. We designed the method for a few selected transcripts whose expression patterns are important for detecting breast cancer cells, and optimized the method for single cell analysis. We successfully detected cells in human blood samples and applied the method to single cells, confirming the heterogeneity of a cell population. / Människokroppen är uppbyggd av miljarder celler och nästan alla innehåller samma arvsmassa. Trots detta finns det många olika celler med olika funktioner vilket är en följd av vilken del av arvsmassan som cellerna använder, dvs vilka RNA-molekyler som finns i varje cell. Den snabba utvecklingen av sekvenseringstekniker har gjort det möjligt att studera när, var och hur varje RNA-molekyl är uttryckt och att få en djupare förståelse för hur människans celler fungerar. Arbetet som presenteras i denna avhandling fokuserar på analys av RNA-molekyler i människans celler. I artikel I beskriver vi en automatiserad metod för att förbereda cellprov för RNA-sekvensering. Det automatiserade protokollet jämfördes med det manuella protokollet, och vi visade att det automatiserade protokollet överträffade det manuella när det gällde provkapacitet samtidigt som en höga reproducerbarheten behölls. I artikel II undersökte vi effekterna som RNA-molekyler från en del av cellen (cellkärnan) har på den totala mängden uttryckta RNA-molekyler. Vi jämförde RNA från hela cellen och från en del av cellen (cytoplasman) och visade att RNA-molekyler med långa och strukturerade 3'- och 5'-otranslaterade regioner och RNA-molekyler med långa proteinkodande sekvenser tenderade att hållas kvar i cellkärnan till en högre grad. Detta resulterade i en ökad komplexitet av RNA-molekylerna i hela cellen, medan vi i cytoplasma-fraktionen lättare kunde hitta de korta och svagt uttryckta RNA-molekyler. I Artikel III och IV studerar vi RNA-molekyler i människans skelettmuskler. I artikel III visar vi att andelen RNA-molekyler uttryckta i skelettmuskler är väldigt lika mellan muskler och mellan olika personer, men att ett stort antal RNA-molekyler var uttryckta i olika nivåer hos kvinnor och män. Artikel IV beskriver RNA-nivåer som svar på upprepade perioder av uthållighetsträning. Artikel V beskriver en metod för att studera ett fåtal utvalda RNA-molekyler. Vi valde RNA-molekyler vars uttryck är viktigt vid analys av bröstcancerceller, och optimerade metoden för analys av enskilda celler. Vi analyserade cancerceller från blodprov och använde metoden för att titta på RNA-nivåer i enskilda celler från en grupp av celler och visade på skillnader i RNA-nivåer inom gruppen. / <p>QC 20150115</p>

This thesis describes the development of microfluidic (MF) platforms for the study of biological and chemical processes. In particular the thesis is divided into two distinct parts: (i) development of a MF methodology to generate tunable cell-laden microenvironments for detailed studies of cell behavior, and (ii) the design and fabrication of MF reactors for studies of chemical reactions. First, this thesis presented the generation of biopolymer microenvironments for cell studies. In the first project we demonstrated a high-throughput MF system for generating cell-laden agarose microgels with a controllable ratio of two different types of cells. The MF co-encapsulation system was shown to be a robust method for identifying autocrine and/or paracrine dependence of specific cell subpopulations. In the second project we studied the effect of the mechanical properties on the behavior of acute myeloid leukemia (AML2) cancer cells. Cell-laden macroscopic agarose gels were prepared at varying agarose concentrations. A modest range of the elastic modulus of the agarose gels were achieved, ranging from 0.62 kPa to 20.21 kPa at room temperature. We observed a pronounced decrease in cell proliferation in stiffer gels when compared to the gels with lower elastic moduli. The second part of the thesis focuses on the development of MF platforms for studying chemical reactions. In the third project presented in this thesis, we exploited the temperature dependent solubility of CO2 in order to: (i) study the temperature mediated CO2 transfer between the gas and the various liquid phases on short time scales, and (ii) to generate bubbles with a dense layer of colloid particles (armoured bubbles). The fourth project involved the fabrication of a multi-modal MF device with integrated analytical probes. The MF device comprised a pH, temperature, and ATR-FTIR probes for in-situ analysis of chemical reactions in real-time. Furthermore, the MF reactor featured a temperature controlled feedback system capable of maintaining on-chip temperatures at flow rates up to 50 mL/hr.

The initial research of this thesis involves the evaluation of different strategies for developing diverse chemistries of highly stable coatings for the automated 96-blade (thin-film) solid phase microextraction (SPME) system. Thin-film geometry increases the volume of extractive phase, and consequently improves the sensitivity of the analysis. Sol-gel technology was used for the preparation of octadecyl (C18)-silica gel thin-film coating. The evaluation of the C18-silica gel SPME extractive phase resulted in stable physical and chemical characteristics and long-term reusability with a high degree of reproducibility. Biocompatible polyacrylonitrile (PAN) polymer was used for the preparation of particle-based extractive phases in order to improve the biocompatible characteristics of SPME coatings for the extraction from biological samples. Three different immobilization strategies were evaluated for developing highly stable coatings for the automated 96-blade SPME system. The spraying was found to be the optimal method in terms of stability and reusability for long-term use. The optimized C18-PAN coating demonstrated improved biocompatibility, stability, and reusability for the extraction of benzodiazepines from human plasma in comparison with those of C18-silica gel coating. To improve the biocompatible properties of the C18-PAN SPME coating for long-term direct analysis from whole blood, different modification strategies were studied and evaluated. The modification of the coating with an extra layer of biocompatible polyacrylonitrile resulted in significant improvement in the blood compatibility in long-term use. ‘Extracted blood spot’ (EBS) sampling was introduced as a novel approach to overcome the limitations of dried blood spot sampling. EBS includes the application of a biocompatible SPME coating for spot sampling of blood or other biofluids. The compatibility of EBS sampling with different analytical methods was demonstrated. The utilization of EBS as a fast sampling and sample preparation method resulted in a significant reduction of matrix effects through efficient sample clean-up. Modified polystyrene-divinylbenzene (PS-DVB)-PAN and phenylboronic acid (PBA)-PAN 96-blade SPME coatings were developed and evaluated for the extraction of analytes in a wide range of polarity. These coatings demonstrated efficient extraction recovery for both polar and non-polar groups of compounds, and presented chemical and mechanical stabilities and reproducible extraction efficiencies for more than 100 usages in biological sample.

Breast cancers with overexpression or amplification of the HER2 tyrosine kinase receptor are more aggressive, resistant to chemotherapy, and associated with a worse prognosis. Currently, these breast cancers are treated with the monoclonal antibody trastuzumab (Herceptin®). Unfortunately, not all patients respond to trastuzumab drug therapy; some patients show de novo resistance, while others acquire resistance during treatment. This thesis describes our RNAi studies to identify novel regulators of the HER2 signaling pathway in breast cancer. Three kinome-wide siRNA screens were performed on five HER2 amplified and seven HER2 non-amplified breast cancer cell lines, two normal breast cell lines, as well as two HER2-positive breast cancer cell lines with acquired trastuzumab resistance and their isogenic trastuzumab-sensitive controls. To understand the main kinase drivers of HER2 signaling, we performed a comprehensive screen that selected against growth inhibitors of the non-HER2 amplified breast cancer cell lines. This screen identified the loss of the HER2/HER3 heterodimer as the most prominent selective inhibitor of HER2-amplified breast cancers. In a trastuzumab sensitization screen on five trastuzumab-treated breast cancer cell lines, we identified several siRNA against the PI3K pathway as well as various other signaling pathways that inhibited proliferation. Finally, in a screen for acquired trastuzumab resistance, PKCη and its downstream targets were identified. Loss of PKCη resulted in a decrease in G1/S transition and upregulation of the cyclin dependent kinase inhibitor p27. Initial data suggest that PKCη promotes p27 ubiquitination and degradation. Taken together, these studies provide novel insight into the complex signaling of HER2-positive breast cancers and the mechanisms of resistance to trastuzumab therapy. This work describes how various kinases can modulate cell proliferation, and points to possible novel drug targets for the treatment of HER2-positive breast cancers.

The polymerase chain reaction (PCR) is an extremely powerful tool for viral detection and screening because it can detect specific infectious agents with great sensitivity and specificity. It works by exponentially amplifying a target viral DNA sequence to high enough concentrations through the use of specific reagents and thermal cycling. It has surpassed culture based methods as the gold standard for viral detection because of the increased speed and sensitivity. Microfluidic approaches to PCR have focused on decreasing the time to thermally cycle, the volumes used for reactions, and they have also added upstream and downstream processes that are of benefit for on-chip viral detection. While these improvements have made great strides over commercially available products in terms of speed, cost, and integration, a major limitation that has yet to be explored is the throughput associated with running PCR. Since each PCR reaction relies on primers with a unique annealing temperature to detect specific viral DNA, only a single virus can be screened for at a time. The device presented here uses two infrared laser diodes that are driven identically by the same laser driver to independently thermally cycle two chambers on the same microfluidic chip. Different temperatures are achieved in the two chambers by modulating the radiation reaching one of those chambers with an optical shutter. Closed loop temperature feedback in both chambers is done with a Labview program and thermocouples embedded in the polymer chip. This allows for accurate temperature measurement without inhibiting the reaction. To demonstrate the capabilities of this device, two different reactions were simultaneously amplified successfully on the same device that have annealing temperatures that differ by 15°C.

Antibiotics are defined as chemical substances that inhibit or limit the growth of microorganisms. Since the second world war, antibiotics have been widely used to reduce the morbidity and mortality associated with serious bacterial infections caused by organisms such as Staphylococcus aureus. However, it has become increasingly difficult to treat bacterial infections due to the emergence of antibiotic resistant strains. The first clinical case of drug resistant bacteria was observed in S. aureus in 1947, just four years after the mass production of penicillin. Since then, resistance has been reported to every antibiotic ever employed. According to the Centres for Disease Control and Prevention of the United States, more than 70% of hospital-acquired infections show resistance to at least one commonly used antibiotic. Coupled with the paucity of therapeutic agents in the pipeline, there is now an urgent demand for new antibiotics. One of the strategies employed to combat drug resistant bacteria requires new chemical entities that work through novel drug targets for which there is no pre-existing resistance. This thesis focuses on the essential metabolic enzyme biotin protein ligase (BPL) as one such new drug target. BPL is the enzyme responsible for covalently attaching the cofactor biotin prosthetic group onto the biotin-dependent enzymes such as the carboxylases, decarboxylases and transcarboxylases. Enzymatic biotinylation proceeds via a two-step reaction whereby biotinyl-5'-AMP is synthesized from biotin and ATP before the biotin moiety is transferred onto the side chain of one specific lysine present in the active site of the biotin-dependent enzyme. One example of an important biotin-dependent enzyme is acetyl CoA carboxylase (ACC). ACC catalyzes the first committed step in fatty acid biosynthesis. Through genetic studies, it has been demonstrated that BPL activity is essential for bacterial survival. The aim for this project was to develop a convenient, high-throughput assay to measure BPL activity. This assay would permit 1) quantitative kinetic analysis of ligands and inhibitors and 2) screening of compound libraries for new BPL inhibitors. We propose that BPL inhibitors can be developed into new antibiotic agents. The novel BPL assay was developed employing fluorescence polarization (FP). FP is a light based technique which uses plane polarized light for the detection of tumbling motion of fluorescent molecules in solution. As polarization of the emitted light is relative to the apparent molecular mass of the fluorophore, this technique can be use for quantitation of changes in molecular mass of target molecules. This enabled 1) rapid kinetic analysis, 2) a minimal number of handling steps, 3) no washing steps and 4) automation by robotics. A first generation assay was developed for Escherichia coli BPL using peptide 85-11 that has been shown to be a convenient substrate. Following the BPL reaction, biotinylated peptides will form large molecular mass complexes with avidin. The amount of product could then be quantitated using FP. Here, kinetic analysis of MgATP (Km 0.25 ± 0.01 mM) and biotin (Km 1.45 ± 0.15 μM) binding produced results consistent with published data. We validated this assay with inhibition studies with end products of the BPL reaction, AMP and pyrophosphate, and a compound, biotinol-5'-AMP. Statistical analysis, performed upon both intraassay and interassay results (n = 30), showed the coefficient of variance to be <10% across all data sets. Furthermore, the Z' factors between 0.5 and 0.8 demonstrated the utility of this technology in high-throughput applications. However, the use of peptide 85-11, a substrate specific to E. coli BPL, does limit the application of this methodology to E. coli. In the second generation FP assay, I adapted this technology for S. aureus BPL by employing the biotin domain of S. aureus pyruvate carboxylase. Insertion of a fluorescein label was achieved by first engineering a cysteine residue into the domain by site directed mutagenesis then incubation with fluorescein-5'-maleimide. A series of mutants was created to investigate optimal positioning of the label into the substrate. Furthermore, the minimal size of the functional domain was determined. Our data showed that the placement of the fluorescein label is an important aspect of this project. Using this approach, I identified that a 90 amino acid domain with the label at position 1134 was optimal. Kinetic analysis of ligand binding showed SaBPL had a Km for biotin at 3.29 ± 0.37 μM and Km for MgATP at 66 ± 16.08 μM. This was in good agreement with data obtained from our previous assay measuring ³H-biotin incorporation. Inhibitor studies with pyrophosphate and analogues of biotin and biotinyl-5'-AMP further validated the assay. Various studies have shown cross-species biotinylation activities by a diverse range of BPLs. Therefore, using this methodology with a biotin domain as the substrate potentially provides a convenient assay for all BPLs. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1374330 / Thesis (M.Sc.) -- University of Adelaide, School of Molecular and Biomedical Science, 2009

Antibiotics are defined as chemical substances that inhibit or limit the growth of microorganisms. Since the second world war, antibiotics have been widely used to reduce the morbidity and mortality associated with serious bacterial infections caused by organisms such as Staphylococcus aureus. However, it has become increasingly difficult to treat bacterial infections due to the emergence of antibiotic resistant strains. The first clinical case of drug resistant bacteria was observed in S. aureus in 1947, just four years after the mass production of penicillin. Since then, resistance has been reported to every antibiotic ever employed. According to the Centres for Disease Control and Prevention of the United States, more than 70% of hospital-acquired infections show resistance to at least one commonly used antibiotic. Coupled with the paucity of therapeutic agents in the pipeline, there is now an urgent demand for new antibiotics. One of the strategies employed to combat drug resistant bacteria requires new chemical entities that work through novel drug targets for which there is no pre-existing resistance. This thesis focuses on the essential metabolic enzyme biotin protein ligase (BPL) as one such new drug target. BPL is the enzyme responsible for covalently attaching the cofactor biotin prosthetic group onto the biotin-dependent enzymes such as the carboxylases, decarboxylases and transcarboxylases. Enzymatic biotinylation proceeds via a two-step reaction whereby biotinyl-5'-AMP is synthesized from biotin and ATP before the biotin moiety is transferred onto the side chain of one specific lysine present in the active site of the biotin-dependent enzyme. One example of an important biotin-dependent enzyme is acetyl CoA carboxylase (ACC). ACC catalyzes the first committed step in fatty acid biosynthesis. Through genetic studies, it has been demonstrated that BPL activity is essential for bacterial survival. The aim for this project was to develop a convenient, high-throughput assay to measure BPL activity. This assay would permit 1) quantitative kinetic analysis of ligands and inhibitors and 2) screening of compound libraries for new BPL inhibitors. We propose that BPL inhibitors can be developed into new antibiotic agents. The novel BPL assay was developed employing fluorescence polarization (FP). FP is a light based technique which uses plane polarized light for the detection of tumbling motion of fluorescent molecules in solution. As polarization of the emitted light is relative to the apparent molecular mass of the fluorophore, this technique can be use for quantitation of changes in molecular mass of target molecules. This enabled 1) rapid kinetic analysis, 2) a minimal number of handling steps, 3) no washing steps and 4) automation by robotics. A first generation assay was developed for Escherichia coli BPL using peptide 85-11 that has been shown to be a convenient substrate. Following the BPL reaction, biotinylated peptides will form large molecular mass complexes with avidin. The amount of product could then be quantitated using FP. Here, kinetic analysis of MgATP (Km 0.25 ± 0.01 mM) and biotin (Km 1.45 ± 0.15 μM) binding produced results consistent with published data. We validated this assay with inhibition studies with end products of the BPL reaction, AMP and pyrophosphate, and a compound, biotinol-5'-AMP. Statistical analysis, performed upon both intraassay and interassay results (n = 30), showed the coefficient of variance to be <10% across all data sets. Furthermore, the Z' factors between 0.5 and 0.8 demonstrated the utility of this technology in high-throughput applications. However, the use of peptide 85-11, a substrate specific to E. coli BPL, does limit the application of this methodology to E. coli. In the second generation FP assay, I adapted this technology for S. aureus BPL by employing the biotin domain of S. aureus pyruvate carboxylase. Insertion of a fluorescein label was achieved by first engineering a cysteine residue into the domain by site directed mutagenesis then incubation with fluorescein-5'-maleimide. A series of mutants was created to investigate optimal positioning of the label into the substrate. Furthermore, the minimal size of the functional domain was determined. Our data showed that the placement of the fluorescein label is an important aspect of this project. Using this approach, I identified that a 90 amino acid domain with the label at position 1134 was optimal. Kinetic analysis of ligand binding showed SaBPL had a Km for biotin at 3.29 ± 0.37 μM and Km for MgATP at 66 ± 16.08 μM. This was in good agreement with data obtained from our previous assay measuring ³H-biotin incorporation. Inhibitor studies with pyrophosphate and analogues of biotin and biotinyl-5'-AMP further validated the assay. Various studies have shown cross-species biotinylation activities by a diverse range of BPLs. Therefore, using this methodology with a biotin domain as the substrate potentially provides a convenient assay for all BPLs. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1374330 / Thesis (M.Sc.) -- University of Adelaide, School of Molecular and Biomedical Science, 2009

Antibiotics are defined as chemical substances that inhibit or limit the growth of microorganisms. Since the second world war, antibiotics have been widely used to reduce the morbidity and mortality associated with serious bacterial infections caused by organisms such as Staphylococcus aureus. However, it has become increasingly difficult to treat bacterial infections due to the emergence of antibiotic resistant strains. The first clinical case of drug resistant bacteria was observed in S. aureus in 1947, just four years after the mass production of penicillin. Since then, resistance has been reported to every antibiotic ever employed. According to the Centres for Disease Control and Prevention of the United States, more than 70% of hospital-acquired infections show resistance to at least one commonly used antibiotic. Coupled with the paucity of therapeutic agents in the pipeline, there is now an urgent demand for new antibiotics. One of the strategies employed to combat drug resistant bacteria requires new chemical entities that work through novel drug targets for which there is no pre-existing resistance. This thesis focuses on the essential metabolic enzyme biotin protein ligase (BPL) as one such new drug target. BPL is the enzyme responsible for covalently attaching the cofactor biotin prosthetic group onto the biotin-dependent enzymes such as the carboxylases, decarboxylases and transcarboxylases. Enzymatic biotinylation proceeds via a two-step reaction whereby biotinyl-5'-AMP is synthesized from biotin and ATP before the biotin moiety is transferred onto the side chain of one specific lysine present in the active site of the biotin-dependent enzyme. One example of an important biotin-dependent enzyme is acetyl CoA carboxylase (ACC). ACC catalyzes the first committed step in fatty acid biosynthesis. Through genetic studies, it has been demonstrated that BPL activity is essential for bacterial survival. The aim for this project was to develop a convenient, high-throughput assay to measure BPL activity. This assay would permit 1) quantitative kinetic analysis of ligands and inhibitors and 2) screening of compound libraries for new BPL inhibitors. We propose that BPL inhibitors can be developed into new antibiotic agents. The novel BPL assay was developed employing fluorescence polarization (FP). FP is a light based technique which uses plane polarized light for the detection of tumbling motion of fluorescent molecules in solution. As polarization of the emitted light is relative to the apparent molecular mass of the fluorophore, this technique can be use for quantitation of changes in molecular mass of target molecules. This enabled 1) rapid kinetic analysis, 2) a minimal number of handling steps, 3) no washing steps and 4) automation by robotics. A first generation assay was developed for Escherichia coli BPL using peptide 85-11 that has been shown to be a convenient substrate. Following the BPL reaction, biotinylated peptides will form large molecular mass complexes with avidin. The amount of product could then be quantitated using FP. Here, kinetic analysis of MgATP (Km 0.25 ± 0.01 mM) and biotin (Km 1.45 ± 0.15 μM) binding produced results consistent with published data. We validated this assay with inhibition studies with end products of the BPL reaction, AMP and pyrophosphate, and a compound, biotinol-5'-AMP. Statistical analysis, performed upon both intraassay and interassay results (n = 30), showed the coefficient of variance to be <10% across all data sets. Furthermore, the Z' factors between 0.5 and 0.8 demonstrated the utility of this technology in high-throughput applications. However, the use of peptide 85-11, a substrate specific to E. coli BPL, does limit the application of this methodology to E. coli. In the second generation FP assay, I adapted this technology for S. aureus BPL by employing the biotin domain of S. aureus pyruvate carboxylase. Insertion of a fluorescein label was achieved by first engineering a cysteine residue into the domain by site directed mutagenesis then incubation with fluorescein-5'-maleimide. A series of mutants was created to investigate optimal positioning of the label into the substrate. Furthermore, the minimal size of the functional domain was determined. Our data showed that the placement of the fluorescein label is an important aspect of this project. Using this approach, I identified that a 90 amino acid domain with the label at position 1134 was optimal. Kinetic analysis of ligand binding showed SaBPL had a Km for biotin at 3.29 ± 0.37 μM and Km for MgATP at 66 ± 16.08 μM. This was in good agreement with data obtained from our previous assay measuring ³H-biotin incorporation. Inhibitor studies with pyrophosphate and analogues of biotin and biotinyl-5'-AMP further validated the assay. Various studies have shown cross-species biotinylation activities by a diverse range of BPLs. Therefore, using this methodology with a biotin domain as the substrate potentially provides a convenient assay for all BPLs. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1374330 / Thesis (M.Sc.) -- University of Adelaide, School of Molecular and Biomedical Science, 2009