The mitochondria are the primary energy providers for most eukaryotic cells. The substrate and products of the mitochondria need to be translocated across the semi-permeable mitochondrial outer membrane (MOM). Mitochondrial porin is an aqueous channel in the MOM thought to provide the primary pathway for metabolite translocation. Porin is a nuclear encoded protein and therefore needs to be transported to the mitochondria, translocated across and assembled within the MOM. Of all the recognition signals required for successful transport, import and assembly, only the β-sorting signal used in assembly is known. In addition, this protein possesses the ability to gate, and in doing so can preferentially allow the passage of anions in the open state and cations in the closed state. However, the precise mechanism by which gating of porin occurs and a complete understanding of porin’s function in vivo remains elusive. The essentiality of porin was examined by constructing a strain of Neurospora crassa deficient for porin. This strain, denoted as WS004, exists as evidence that porin is non-essential for the survival of Neurospora crassa. However, the loss of porin results in a reduction in growth rate due to the dysfunction of the cytochrome mediated respiratory pathway, which was made evident by the reduction of cytochrome b and almost complete lack of cytochrome aa3. WS004 survives by inducing the expression of alternative oxidase, which funnels the electrons from the Q pool directly to oxygen, bypassing the cytochrome b and aa3 containing complexes III and IV respectively. Additional phenotypic differences observed included loss in ability to produce aerial hyphae, reduced amount of conidia produced and strains that were female sterile. It was determined, that additional genetic factors influenced the resulting phenotype due to the loss of porin. LC-MS/MS, in combination with iTRAQ labelling, was utilized to examine changes in the proteome profiles of porin containing and porin lacking mitochondria and showed several different proteins as significantly up- or down-regulated which lend to an explanation to some of the phenotypes observed. Taken together, these results demonstrate the central role of porin in regulating both mitochondrial and cellular processes.

The thesis describes development and application of coupled resonant coil sensors, which is of growing interest for remote monitoring applications. An interrogation technique, which improves the accuracy and interrogation range of coupled resonant coil sensors, is introduced. The method uses time-domain gating to produce measurements that are dominated by the response of the sensor coil and are immune to surrounding object interference. For application in structural health monitoring a low cost embeddable coupled coil sensor, which is able to monitor the corrosion potential of reinforcement steel is presented. Results of an accelerated corrosion test using the sensor indicate that corrosion potential can be monitored with a resolution less than 10 mV and a sensitivity of 0.76 kHz/mV. The last part describes a coupled-coil pH sensor based on pH electrode potential measurement. A linear response over a 4 to 10 pH dynamic range and 50 kHz/pH sensitivity are achieved with a 0.1 pH resolution and 30 s response time.

Hyperthermophilic proteins are of great interest in both the academic and industrial world in understanding how these proteins are capable of retaining their biological activity under such harsh environmental conditions. This thesis studies a tetrabrachion stalk domain from Staphylothermus marinus, know as Right Handed Coiled Coil (RHCC). This protein is of interest due to its extreme thermostability and its affinity for heavy metals. We aim to better understand the reason for the extreme thermal stability of the protein and to take advantage of the proteins affinity for heavy metals with a view to developing a novel approach to bioremediate Hg2+, a major environmental pollutant. Our results clearly indicated that the protein is more thermostable in alkaline conditions in comparison to acidic conditions. This observation can be explained by careful inspection of the high resolution structure. Our data also clearly show that RHCC is able to bind ionic mercury compounds such as mercury nitrate and dipotassium mercury iodide.

Die zum Fremdsprachenerwerb wesentliche Vermittlung interkultureller Kommunikationskompetenzen wird in den verfügbaren Lehrplänen in Manitoba für Deutsch als Fremdsprache vorausgesetzt. Eine Umfrage unter den Deutschlehrern sowie repräsentative Interviews zeigen, dass interkulturelle Kommunikationskompetenzen in der Praxis oft nicht ihrem Wichtigkeitsgrad entsprechend in den Unterricht integriert werden. Der Erfolg des DaF-Programms in Manitoba hängt dementsprechend von der Sensibilisierung der Lehrer sowie einer Reform und Modernisierung von erforderlichen Prüfungsmöglichkeiten und Qualitätssicherung der Vermittlungsinhalte ab.

DLX transcriptional targets have been implicated during retinal development. The Crx (Cone-Rod homeobox) gene is required for the differentiation and maintenance of cone and rod photoreceptors. Otx2 (Orthodenticle homeobox 2) is a key regulator of photoreceptor cell fate. The Dlx1/Dlx2 mutant mouse retina has a significant reduction of retinal ganglion cells with aberrant Crx expression in the neuroblastic layer and increased retinal Otx2 expression. We hypothesized that the Dlx homeobox genes directly repress Crx and Otx2 expression during retinal development. Expression of CRX demonstrates increased transcript and protein expression in the Dlx1/Dlx2 double knockout retina at E18.5, suggesting that these DLX transcription factors may repress CRX expression. OTX2 expression is increased in the Dlx1/Dlx2 knockout retina at E16.5 suggesting that DLX2 negatively regulates OTX2 expression. The Dlx1/Dlx2 knockout has aberrant and ectopic expression of CRX in the retina along with increased OTX2 expression. Our data suggests that both CRX and OTX2 are transcriptional targets directly repressed by the DLX1 and DLX2.

Eating disorders are a significant health concern due to their high rates of comorbidity, mortality, and the physical and mental distress they cause. While many people are aware of the negative effects eating disorders have on physical and mental health, few realize that suicide is a potential outcome. Although the relationships between eating disorders and suicidality have been examined in clinical populations with anorexia nervosa and bulimia nervosa, these relationships have not been examined in the general population, nor with binge eating disorder. This study aimed to investigate these relationships in the Collaborative Psychiatric Epidemiologic Surveys (CPES, N=20,013), a large, nationally representative sample of adults in the United States. Logistic regression analyses revealed that individuals with a history of each eating disorder reported higher rates of suicide ideation and suicide attempts. Rates of suicide experiences among this population are similar to individuals with a history of major depression and a comorbid anxiety disorder. It is clear from this study that suicide remains a significant concern for individuals suffering from an eating disorder in the general population. It is essential that clinicians screen for suicide experiences in individuals suspected of suffering from an eating disorder.

The purpose of this research was to develop a model for the implementation and evaluation of citizen-user involvement in mental health policymaking. The study explored the pathways through which the experiential knowledge of citizen-users enters policy processes, how the outcomes of citizen-user involvement are conceptualized by policy actors, and the contextual factors that influence the implementation and outcomes of involvement. Qualitative instrumental case study methodology was used to focus on the policy field of mental health and social housing. Data were collected through key informant interviews with a purposive sample of 21 people recruited from four policy actor groups: citizen-users, representatives of advocacy organizations, government officials (elected representatives and bureaucrats), and service providers. A review of policy documents as well as forum, committee and task force reports provided additional data for the study. The research built on a previous study that developed a model of the important processes that are used to engage citizen-users in decisions about services and policies. Findings from the current study built on this model by exploring the outcomes, pathways and contexts of involvement from the perspectives of policy actors. Four categories of outcomes were identified: substantive, instrumental, normative and personal. Benefits and risks of involvement were identified within each category. Participants described direct and indirect pathways through which the voices of citizen-users have gained access to policymaking. Direct pathways were those through which individual and collective voices of citizen-users have communicated directly with decision makers. Indirect pathways represented the ways in which the voices of citizen-users were mediated by other policy actors. The findings also highlighted five contextual factors that have influenced citizen-user involvement: the socio-political environments, institutional characteristics, participant characteristics, opportunities to be involved and other influences on policymaking. This research has added to knowledge about the important components of citizen-user involvement in policymaking. The results provide guidance to policy actors about ways to enhance involvement. People with mental health and social housing needs have important experiential and other knowledge to contribute to policymaking. The challenge is the ongoing search for the means to ensure that their voices are heard and carry weight.

Reliable determinations of ecosystem scale fluxes of net carbon (C) and greenhouse gases for northern peatland ecosystems are of great value to determine the impact of soil warming and altered precipitation on emissions. Additionally, few studies have been performed which measure the C fluxes, particularly methane flux (FCH4), during the spring melt and fall freeze up periods, therefore making it difficult to provide adequate annual C estimates from northern peatland ecosystems. This study aimed to determine ecosystem scale FCH4 from a eutrophic Subarctic fen at Churchill, Manitoba (58°45'N 94°4'W), to understand (a) seasonal trends over two consecutive growing seasons,(b) if over-winter stored CH4 was released as a pulse during the spring-melt period, and (c) soil temperature - FCH4 relations for modelling FCH4 over the spring-melt period. An ecosystem scale methane (CH4) and carbon dioxide (CO2) flux measurement system using the eddy covariance (EC) technique was used from late-June to mid-October of 2008 and early-June to late-September of 2009, with focus on the spring-melt period of late-May to mid-July of 2009. The EC flux measurement system consisted of a closed-path RMT-200 Fast Methane Analyzer (Los Gatos Research Inc.) along with a LI-7500 open-path CO2/H2O gas analyzer (LI-COR Biosci.) and a CSAT3 3-dimensional sonic anemometer (Campbell Sci.). The system was powered by a combination of wind, solar, and gas electric generation. The EC flux measurement system provided seasonal FCH4 values of 0 – 90 nmol CH4 m-2 s-1, similar to previous studies in Subarctic and Arctic peatlands which incorporated the EC technique. A melt period CH4 emission burst was not observed, rather a gradual increase in emission over the spring period. Modelled FCH4 using a temperature-response curve relationship with soil temperature at 5 cm depth over the spring-melt period (May 30 – July 19, 2009) showed the fen to be a net source of CH4, of 1.4 mmol m-2 CO2 equivalent.

Chronic pain is estimated to affect approximately 29% of the Canadian population and causes far-reaching lifestyle changes and difficulties that have personal, social, and economic impact. Managing pain is a complex task that recently has been recognized as requiring a multidisciplinary approach. Significant amounts of research have been conducted to understand the biological and cognitive bases of pain; and yet, spiritual aspects of chronic pain have received less attention. Despite this, spirituality remains an important coping strategy as reported by individuals managing chronic health conditions. Some interventions in use today have spiritual roots; however, western medical treatments of pain often do not recognize or utilize spirituality on a regular basis. Utilizing one such intervention, namely mindfulness meditation (MM), this study attempted to understand the role that MM plays in the management of chronic pain and the development of spirituality. This study measured self-reported pain, anxiety, depression, and spirituality in individuals engaged in an 8-week group process in which individuals were provided chronic pain related psycho-education either with MM or without. A repeated measures model was utilized to measure changes that occurred in reported pain experiences (using the McGill Pain Questionnaire) and in spirituality (using the Spiritual Assessment Scale ) to determine if changes could be associated with the teaching of MM. Differences in Affective pain ratings were observed and correlations between pain ratings and spirituality discussed. Reasons for the lack of significant findings, including sample size, power, and measures are discussed.

This thesis describes the production of adenovirus-based vaccines containing codon-optimized genes from Nipah virus and Crimean-Congo Hemorrhagic Fever virus. Genes encoding envelope proteins from Crimean-Congo Hemorrhagic Fever Virus and Nipah Virus were codon-optimized for translation in human cells and constructed using a modified method of non-gapped gene synthesis, while the entire M segment encoding the glycoprotein precursor for Crimean-Congo Hemorrhagic Fever Virus was commercially synthesized. Genes were cloned into recombinant human adenovirus serotype 5 and the resulting viral particles were amplified, titred and analyzed for in vivo efficacy. Results show that a modified method of non-gapped gene synthesis is an effective and efficient method of producing antigen-encoded DNA and at a fraction of the cost and time required for commercial synthesis. Furthermore, adenovirus-based vaccines induce both cellular and humoral immune responses providing for a highly efficacious vaccine during potential disease outbreaks, where time to completion is of utmost importance. This study has shown that recombinant adenoviral vaccines for Crimean-Congo Hemorrhagic Fever virus and Nipah virus can be produced rapidly and efficiently from virtual DNA sequence to optimized recombinant vaccines in just eight months.