Life Sciences Commons^™

Atpase Regulation In The Maltose Transporter, Alister D. Gould

Electronic Thesis and Dissertation Repository

This thesis investigates the mechanism of activity-coupling in the maltose transporter of Escherichia coli (MalFGK₂); the way ATP hydrolysis is prevented in the absence of maltose, and then enabled to drive maltose transport. Like other ATP binding cassette importers, MalFGK₂ requires substrate to be presented by a peripheral substrate-binding protein, in this case the maltose binding protein (MBP). MBP predominantly adopts an ‘open’ resting state, but undergoes a rotation of its two domains to a ‘closed’ state after maltose binding. In the closed state MBP is able to activate MalFGK₂ to stimulate ATP hydrolysis and maltose …

Septal Modulation Of The Hippocampus, Siew Kian Tai

Electronic Thesis and Dissertation Repository

The medial septum (MS) is the main source of acetylcholine to the hippocampus, a structure involved in memory and Alzheimer’s disease (AD). Learning and memory involve long-term changes in synaptic strengths, and are suggested to be facilitated by a brain wave, theta rhythm in the hippocampus. Since medial septal neurons influence hippocampal neural activity, lesion of two neuronal populations in the MS, cholinergic and GABAergic, was performed by intraseptal infusion of 192 IgG-saporin and orexin-saporin, respectively. I hypothesized that 1) activation of cholinergic cells by vestibular stimulation induces an atropine-sensitive theta rhythm, modulates synaptic transmission and enhances long-term potentiation (LTP), …

Stable Isotope Analyses Of Bat Fur: Applications For Investigating North American Bat Migration, Erin E. Fraser

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Many aspects of North American bat migration are not well documented. Stable isotope analyses of animal tissues can elucidate migratory origin, but this technique has not been widely applied to bats. This dissertation i) uses fur stable isotope analyses to investigate North American bat migration and ii) highlights some of the strengths and weaknesses of this analytical technique when applied to bat systems. I conducted stable hydrogen, carbon, and nitrogen isotope analyses on fur from five bat species.

I documented δD_fur heterogeneity in summer resident populations of Myotis lucifugus, Lasiurus borealis, Lasiurus cinereus, and Lasionycteris noctivagans. Stable …

Planning, Implementation, And Formative Evaluation Of A Food Literacy Program, Heather M. Thomas

Electronic Thesis and Dissertation Repository

The overall purpose of this dissertation was to examine the planning, implementation, and formative evaluation of a community-based food literacy program for youth. Article 1 provided a summary of the community-based cooking program for at-risk youth. Objectives included the provision of applied food literacy and cooking skills education augmented with fieldtrips to local farms. Eight at-risk youth (five girls and three boys, mean age = 14.6) completed the intervention. Post intervention, five of eight participants completed in-depth interviews about their experiences.

Article 2 was a formative evaluation of the cooking program focused on gaining an understanding of participants’ (i.e., at-risk …

Overwintering Energetics Of Lepidoptera: The Effects Of Winter Warming And Thermal Variability., Caroline M. Williams

Electronic Thesis and Dissertation Repository

Winter temperatures are changing rapidly, and although winter warming reduces cold stress for overwintering ectotherms, temperature-mediated increases in metabolic rate can decrease fitness in dormant insects by increasing consumption of energy reserves. Increases in thermal variability also increase energetic demands, due to non-linear thermal response curves. My objective was to quantify the negative effects of winter warming and increases in thermal variability on a range of Lepidopteran species. As overwintering insects rely on lipid catabolism, accurate lipid measurement is central to my dissertation; so I first compared four methods of lipid quantification; concluding thin layer chromatography was the only method …

The Intracellular Behaviour Of Burkholderia Cenocepacia In Murine Macrophages, Jennifer S. Tolman

Electronic Thesis and Dissertation Repository

Burkholderia cenocepacia is an opportunistic pathogen causing life-threatening infections in cystic fibrosis and other immunocompromised patients. The bacterium survives within macrophages by interfering with typical endocytic trafficking, resulting in delayed maturation of a B. cenocepacia-containing phagosome. We hypothesize that B. cenocepacia alters gene expression after internalization by macrophages, inducing genes involved in intracellular survival and host adaptation. Furthermore, we hypothesize that specialized bacterial secretion systems are involved in the interactions between intracellular bacteria and macrophages. In this work, we characterize later-stage infection of macrophages by B. cenocepacia, showing replication within an acidified endosomal compartment suggestive of a phagolysosome. …

Morpho-Physiological Analysis Of Interneuronal Populations In The Rat Piriform Cortex Before And After Kindling Induced Epilepsy, Cezar Gavrilovici

Electronic Thesis and Dissertation Repository

The piriform cortex (PC) is involved in olfactory sensory processing, associative learning tasks and is highly seizurogenic. Understanding how interneurons participate in these behaviours, especially their contribution in epileptogenic mechanisms, is hampered by an incomplete understanding of their functional and morphological diversity. The hypothesis in this work is that kindling-induced epilepsy alters the firing properties of PC interneuronal populations. Altered/impaired interneuronal firing could lead to abnormal processing in the PC and epileptogenesis. Therefore it was important to first identify and describe interneuronal morpho-functional properties in the unkindled brain and then to assess the electrophysiological parameters following kindling.

Based on interneuronal …

Structure And Dynamics Of The Membrane Protein Bacteriorhodopsin Studied By Mass Spectrometry, Yan Pan

Electronic Thesis and Dissertation Repository

Membrane proteins continue to represent a major challenge for most analytical techniques. Using bacteriorhodopsin (BR) as model system, this work aims to develop mass spectrometry (MS)-based approaches for exploring the structure, dynamics and folding of membrane proteins.

As the first step, BR in its native lipid environment was exposed to hydroxyl radicals, which were produced by laser photolysis of hydrogen peroxide. It was found that the resulting methionine (Met) labeling pattern was consistent with the known BR structure. This finding demonstrates that laser-induced oxidative Met labeling can provide structural information on membrane proteins. In subsequent experiments, the effects of different …

Brassinosteroid-Mediated Stress Tolerance In Arabidopsis Thaliana, Tawhidur Rahman

Electronic Thesis and Dissertation Repository

Brassinosteroids (BRs) are a group of steroidal plant hormones that are essential for proper plant development and also promote stress tolerance. Without BRs, plants are dwarfs and infertile. To understand the molecular mechanisms underlying BR-mediated stress tolerance, global gene expression analysis of untreated and 24-epibrassinolide (EBR)-treated Arabidopsis thaliana seedlings under non-stress and heat stress (HS) conditions was carried out. Microarray data analysis indicated that stress-related genes were predominant within the EBR up-regulated gene data set. Furthermore, several of these genes were abscisic acid (ABA) and jasmonic acid (JA) related. Measurements of endogenous hormones showed significant increases in the levels ABA …

Effects Of Methamphetamine On Sexual Behavior, Karla S. Frohmader

Electronic Thesis and Dissertation Repository

Methamphetamine (Meth) is a highly addictive psychostimulant associated with enhanced sexual desire, arousal, and sexual pleasure. Moreover, Meth abuse is frequently linked with the practice of sexual risk behavior and increased prevalence of Human Immunodeficiency Virus (HIV). Currently, the neurobiological basis for this drug-sex nexus is unknown. Moreover, there is a lack of studies investigating the effects of Meth on sexual behavior and more importantly, compulsive sex-seeking behavior, under controlled experimental settings in animal models. First, using immuhistochemistry for mating- and Meth-induced neural activation it was demonstrated that Meth administration in male rats activates neurons in brain regions of the …

Structural Characterization Of Hip2 Enzyme Interactions In Ubiquitination, Benjamin W. Cook

Electronic Thesis and Dissertation Repository

The ubiquitin proteolysis pathway utilizes three enzymes, an E1 activating enzyme, an E2 conjugating enzyme and an E3 ligating enzyme, to respectively activate, transfer and ligate ubiquitin (Ub) onto a substrate protein. The creation of a K48-linked poly-Ub chain on a substrate will target this protein to be degraded by the 26S proteosome. E2 conjugating enzymes are central proteins in this pathway and interact with the E1 and E3 enzymes to perform Ub transfer. The mechanism by which Ub molecules are interconnected remains poorly understood. The E2 enzymes HIP2 and Ubc1 have been shown to create poly-Ub chains in the …

Critical Stages In Viral Replication - Entry, Gene Regulation And Egress, Matthew S. Miller

Electronic Thesis and Dissertation Repository

Despite the amazing biological diversity exhibited by viruses, their very existence relies upon their ability to overcome a set of common barriers. The nature of these barriers reflects the nature of viruses themselves. During their extracellular phase, viruses are metabolically inert obligate parasites. Upon encountering a host cell, productive infection necessitates that the virus successfully enter the cell, regulate the expression of its genes, and after assembling new progeny particles, egress such that the cycle of infection can continue. These three basic processes are not only attractive candidates for therapeutic intervention, but also reveal much about virus biology in the …

Structure-Function Analysis Of Enzymes Of The Polyisoprenyl-Phosphate Hexose-1-Phosphate Transferase Family, Kinnari B. Patel

Electronic Thesis and Dissertation Repository

Enzymes of the polyisoprenyl-phosphate hexose-1-phosphate transferase (PHPT) family are integral membrane proteins that initiate the synthesis of glycans by catalyzing the transfer of a hexose-1-phosphate sugar from UDP-hexose to the lipid carrier undecaprenyl phosphate (Und-P). These glycans such as O antigen and exopolysaccharide (EPS) provide bacteria with protection and adaptation to the environment and host immune factors. The role of PHPT proteins in initiation and the absence of any eukaryotic homologues make them an attractive target for novel antimicrobials; however study of these proteins is difficult due to the presence of multiple transmembrane helices. A requirement of the C-terminal domain …

Trace Metal Limitation And Its Role In Oxidative Stress Of Coral Algal Symbionts; Implications For Thermally Induced Coral Bleaching Events., Katrina Lynn Iglic

Electronic Thesis and Dissertation Repository

Coral bleaching, the process in which corals expel their photosynthetic symbionts (Genus Symbiodinium), is caused by high temperature and irradiance stress. The synergistic effect of each stressor is the generation of damaging reactive oxygen and nitrogen species (ROS and RNS), which are normally mitigated physiologically by antioxidant enzyme activity. Antioxidant enzymes require iron, copper, manganese and zinc in their structure and the limited nature of these trace metals in reef environments may enhance ROS production under elevated temperature and irradiance. The hypothesis tested within this thesis was that a limited availability of the trace metals Fe, Mn, Cu and …

A Model System In Landscape Genetics – The Insect Inhabitants Of Pitcher Plants, Gordana Gojko Rasic

Electronic Thesis and Dissertation Repository

Landscape genetics is a rapidly growing field that investigates how landscape and environmental features interact with microevolutionary processes to give rise to spatial genetic variation in populations. The ability to predict landscape effects on genetic patterns has been limited by the lack of studies conducted on more than one species, over multiple spatial scales and in replicated landscapes. A system that allows for such studies are the insect inhabitants of the purple pitcher plant (Sarracenia purpurea): the pitcher plant flesh fly (Fletcherimyia fletcheri), the midge (Metriocnemus knabi) and the mosquito (Wyeomyia smithii). In this thesis, I worked towards developing this …

Purine Transport And Metabolism In Microvascular Endothelial Cells, Derek B J Bone

Electronic Thesis and Dissertation Repository

The microvascular endothelium serves as the barrier between the blood and perfused tissues. Proper function of the endothelium is dependent on the ability of the endothelial cells to produce nitric oxide and form tight junctions between themselves. Dysfunction occurs when stresses overwhelm the endothelial cell, with oxidative stress being a leading cause. Intracellular metabolism of purine nucleosides and nucleobases has been implicated in the production of oxidative stress. Nucleosides (e.g. adenosine) and nucleobases (e.g. hypoxanthine) are moved across cell membranes by a specialized family of proteins called equilibrative nucleoside transporters (ENTs).

We characterized primary human cardiac microvascular endothelial cells (MVECs) …

Exploring Bacterial Nanowires: From Properties To Functions And Implications, Kar Man Leung

Electronic Thesis and Dissertation Repository

The discovery of electrically conductive bacterial nanowires from a broad range of microbes provides completely new insights into microbial physiology. Shewanella oneidensis strain MR-1, a dissimilatory metal-reducing bacterium, produces extracellular bacterial nanowires up to tens of micrometers long, with a lateral dimension of ~10 nm. The Shewanella bacterial nanowires are efficient electrical conductors as revealed by scanning probe techniques such as CP-AFM and STM.

Direct electrical transport measurements along Shewanella nanowires reveal a measured nanowire resistivity on the order of 1 Ω∙cm. With electron transport rates up to 10⁹/s at 100 mV, bacterial nanowires can serve as a …

Decoding Motor Intentions From Human Brain Activity, Jason P. Gallivan

Electronic Thesis and Dissertation Repository

“You read my mind.” Although this simple everyday expression implies ‘knowledge or understanding’ of another’s thinking, true ‘mind-reading’ capabilities implicitly seem constrained to the domains of Hollywood and science-fiction. In the field of sensorimotor neuroscience, however, significant progress in this area has come from mapping characteristic changes in brain activity that occur prior to an action being initiated. For instance, invasive neural recordings in non-human primates have significantly increased our understanding of how highly cognitive and abstract processes like intentions and decisions are represented in the brain by showing that it is possible to decode or ‘predict’ upcoming sensorimotor …

Experiences Of Embodiment: Analysis Of Muslim Women's Participation In Physical Activity, Natalie V.K Szudy

Electronic Thesis and Dissertation Repository

This dissertation examines the relationship the „lived body‟ experiences of veiled Muslim females in sport and physical activity. When considering the relationship between religious requirements and participation in sport and physical activity, the discussion of the conflicts that have occurred with some veiled Muslim female athletes can support the examination of the articulation of sport and religion. In this dissertation, I will explore the application of logical and philosophical discussion as an analytic framework in the evaluation of social, cultural and religious discourse. The application of this framework will contribute to the evaluation of epistemological premises that have contributed to …

Mitochondrial Metabolic Suppression And Reactive Oxygen Species Production During Hypometabolism In Mammals, Jason Cl Brown

Electronic Thesis and Dissertation Repository

During hibernation, daily torpor, and fasting, mammals reduce metabolic rate (MR) up to 99%, 95%, and 30%, respectively, compared to resting levels. Mitochondrial metabolic suppression likely contributes to this MR reduction, and the first objective of this study was to determine the relative contributions of active, regulated inhibition and passive thermal effects as body temperature (T_b) falls, to mitochondrial metabolic suppression, and to examine the mechanisms involved using top-down elasticity analysis and novel statistical approach. The second objective of this study was to determine how mitochondrial metabolic suppression affects mitochondrial reactive oxygen species (ROS) production, a topic which …

Regulation Of G Protein Signaling By Goloco Motif Containing Proteins, Peishen Zhao

Electronic Thesis and Dissertation Repository

Signal transduction via heterotrimeric G proteins in response to transmembrane G protein-coupled receptors plays a central aspect in how cells integrate extracellular stimuli and produce biological responses. In addition to receptor-mediated activation of heterotrimeric G proteins, during the last few decades, accessory proteins have been found to regulate G protein activity via different mechanisms. Several proteins have been identified that contain multiple G protein regulatory domains. Using various molecular and biochemical approaches, we have characterized the effects of two such proteins, G18 and RGS14, on G protein activity. Both proteins contain a second G protein binding domain in addition to …

Metastatic Disease: Interactions Between Tumor Cells And Host Environment During Cancer Cell Spread, Jennifer M. Maclean

Electronic Thesis and Dissertation Repository

Tumor and metastasis formation are not cell autonomous phenomena, but rather an evolution of disease within and responding to the host environment. Metastatic spread from a primary tumor occurs as a result of a complex interplay between tumor cells and the host, wherein tumor cells must escape the primary tumor, enter the host vasculature, travel to and arrest in a distant tissue and survive and grow in that new organ. It is known that cells that progress through these stages must both escape and exploit host systems, yet the mechanisms used are not fully understood. Therefore, the goal of this …

The Regulation Of Cell Division And Neurogenesis By The Chromatin Remodeling Protein Atrx, Kieran L. Ritchie

Electronic Thesis and Dissertation Repository

Mutations in the ATRX gene cause alpha thalassemia mental retardation X-linked syndrome (ATR-X). Individuals with ATR-X syndrome show a broad range of developmental abnormalities including severe cognitive, behavioural, and learning deficits, seizures, and microcephaly, indicating that ATRX is essential for normal development of the central nervous system. Conditional inactivation of Atrx in the developing mouse forebrain leads to a reduction in cerebral cortical size, elevated levels of p53-dependent neuronal apoptosis, and dysgenesis of certain subcortical structures, including the hippocampus and dentate gyrus, confirming a requirement for Atrx in mammalian brain development. The mammalian ATRX gene encodes a member of the …

Role Of Rac1 In Myocardial Tnf-Alpha Expression In Sepsis, Ting Zhang

Electronic Thesis and Dissertation Repository

Tumor necrosis factor-a (TNF-alpha) is a pro-inflammatory cytokine and its high levels of expression in the heart leads to cardiac dysfunction in sepsis. However, the underlying molecular mechanisms of regulating myocardial TNF-alpha expression are not fully understood. The aim of this thesis was to investigate the role of Rac1 in myocardial TNF-alpha expression and cardiac dysfunction during sepsis. Studies were performed using cultured neonatal cardiomyocytes and a mouse model of endotoxemia.

PI3K-mediated Rac1 activation is required for induction of TNF-alpha mRNA and protein expression in cardiomyocytes and cardiac dysfunction during endotoxemia. Rac1 promotes TNF-alpha mRNA expression via NADPH oxidase/ERK1/2 and …

The Effects Of Superovulation And Embryo Culture On Genomic Imprinting In A Mouse Model System, Brenna A. M. Velker

Electronic Thesis and Dissertation Repository

Genomic imprinting is a specialized transcriptional mechanism resulting in the unequal expression of alleles based on their parent-of-origin. Imprinted genes are critical for embryonic and fetal development and their dysregulation is linked to a group of human diseases called imprinting disorders, including Beckwith-Wiedemann Syndrome, Angelman Syndrome and Silver-Russell Syndrome. Two critical phases of genomic imprinting exist. The acquisition phase occurs in developing germ cells, asynchronously for different imprinted loci, while the maintenance phase takes place during preimplantation development, while the rest of the genome is undergoing demethylation. Increased frequencies of human imprinting disorders are observed in children following the use …

Dissecting The Molecular Role Of Distinct Binding Interfaces On The Retinoblastoma Tumor Suppressor In Growth Control And Tumorigenesis., Matthew J. Cecchini

Electronic Thesis and Dissertation Repository

The retinoblastoma tumor suppressor protein (pRB) functions to maintain proliferative control and act as a barrier to tumorigenesis. pRB is capable of regulating E2F transcription factors to mediate control of proliferation through transcriptional regulation of S-phase target gene expression. In addition, pRB can stabilize the CDK inhibitor p27 through an interaction with two ubiquitin ligase complexes. Further, pRB is capable of forming a unique interaction with E2F1 termed the ‘specific’ interaction that is capable of blocking E2F1 induced apoptosis. These functions of pRB are mediated by distinct binding interfaces and their contributions to the overall functionality of pRB are not …

Extent, Characteristics And Downstream Effects Of Stream Enclosure In Southwestern Ontario, Katie L. Stammler

Electronic Thesis and Dissertation Repository

In a 16000 km² area of southwestern Ontario, almost 15% of all streams have been enclosed (buried) largely for agricultural purposes. ArcGIS was used to characterize the natural features of catchments and to calculate enclosedness (proportion of stream network enclosed; x̅ = 16.5%, n = 10106). Catchments with the highest enclosedness received >990 mm of precipitation annually and had high drainage density (>1.9 km/km²), while catchments with the lowest enclosedness receivedannually, were characterized by clay and undrumlinized till plains and had relatively shallow water tables (m). These natural features influence where enclosure is likely to occur, …

Regional Specification Of The Xenopus Lateral Plate Mesoderm, Steven J. Deimling

Electronic Thesis and Dissertation Repository

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Successful patterning of the embryo, from establishing the three primary axes to the regional specification of tissue progenitors is essential to generating a viable embryo. The three germ layers in the early embryo undergo patterning through slightly different mechanisms. The tissue of interest to this study is the lateral plate mesoderm (LPM), which will give rise to the lineages of the cardiovascular system and is essential for regional specification of adjacent germ layers. …

Transactivation By Human Adenovirus Early Region 1a-Conserved Region Three, Jailal Ng Ablack

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One of the critical functions of human adenovirus (hAd) early region 1A (E1A) protein is to activate transcription of the early viral genes. The largest isoform of E1A contains a unique region termed conserved region 3 (CR3), which includes a Cysteine-4 (C4) zinc finger domain. This region activates viral gene expression by interacting with and recruiting cellular transcription machinery to the regulatory regions of early viral genes. Although this process has been studied at length with hAd type 5 E1A, far less is known about how the E1A proteins from other hAd types activate transcription. There are dramatic differences in …

Serum Iron Uptake And Virulence In Staphylococcus Aureus, Federico C. Beasley

Electronic Thesis and Dissertation Repository

The high affinity iron scavenging glycoprotein transferrin sequesters trace amounts of serum Fe³⁺ to concentrations below what is required to sustain microbial life. Iron may be liberated from this important innate immune factor after interaction with molecules that chelate or reduce Fe³⁺. Organisms with cognate transport systems for these iron coordinating molecules can survive in the bloodstream using transferrin iron.

Staphylococcus aureus is an opportunistic bacterial pathogen. S. aureus executes numerous strategies for overcoming the innate immune barrier of iron deprivation in the bloodstream. In addition to specialized mechanisms for hemoglobin iron extraction, S. aureus can proliferate …