Graduate Theses & Dissertations

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Changes in Forms of Uranium in Anoxic Lake Sediments and Porewaters Near an Abandoned Uranium Mine, Bancroft, Ontario
Soluble uranium (U) has been observed continuously in the porewaters of Bentley Lake, a lake with semi-permanent anoxic sediments, despite the fact that reduced U(IV) is known to be insoluble. To be able to predict the fate and mobility of U that has been deposited in lake sediments, it is very important to understand the factors that determine soluble uranium in anoxic environments. Understanding soluble U species is crucial for predicting its behavior in natural systems as well as for the development of U remediation schemes. To explore the factors affecting soluble U in natural environments, anoxic lake sediments and porewaters were tested using two analytic methods, ICP-MS and ESI-HR-MS. Reduced uranium (U(IV)) can be precipitated as U(IV)-NdF3. Using this method revealed that most of the uranium in porewater is not able to be co-precipitated with NdF3. In addition, UO2+ was found using ESI-HR-MS, showing uranyl ions exist in reduced porewater. However, the UO2+ might be attached to some organic groups rather than present as free ions. Seasonal variation and air exposure experiments on the mobility of U between sediments and porewater were observed to test for changes of the redox state of U as a function of sample collection and storage. The results of this study will contribute to better remediation strategies for U tailings and will help U mining operations in the future. Author Keywords:
Changes in the Representations of Women from the 1980s Turkish Cinema to New Turkish Cinema
The thesis examines the changes in the representations of women in Turkish cinema from the 1980s through the 2000s in terms of semiotic codes. To demonstrate the shifts of the representations of women over three decades, four films directed by two representative filmmakers of the 1980s and the 2000s are analyzed within the context of gender codes: A Sip of Love (Atıf Yılmaz/1984), The Night, Angel and Our Gang (Atıf Yılmaz/1994), The Third Page (Zeki Demirkubuz/1999) and Envy (Zeki Demirkubuz/2009). Using gender as code in deconstructing women’s characteristics and their representations in the films, the research highlights the structural image of women in the semiotic sense. The stereotypical representations in Turkish cinema question social norms, patriarchy, and Islamic norms in Turkey. Author Keywords: Gender codes, Islam, Patriarchy, Semiotics, The representations of women , Turkish cinema
Characterisation of the Giardia Tata-Binding Protein - Preparation for an in vivo approach
The aim of this work was to identify the DNA sequences recognized by the Giardia TBP (gTBP) in vivo by using a chromatin immunoprecipitation assay (ChIP). Since a specific antibody for the protein of interest is required for this assay, a company was contracted to produce and purify a custom polyclonal antibody from the immunization of rabbits. Recombinant GST-gTBP was produced at a suitable yield and purity and used as the immunogen. The antibody was then tested for reactivity to the native protein in our laboratory. By Western blot analysis, it was possible to observe the enrichment of the gTBP within the nuclear fraction compared to a cytoplasmic fraction extracted from Giardia cells. However, the antibody could not be successfully used in an immunoprecipitation assay - suggesting that the antibody is unable to bind to the native structure of gTBP. Therefore, the focus of this work was changed to analyse gTBP via multiple sequence alignments, homology modelling and BLAST to identify any unique regions that may contribute to its unusual binding characteristics. These techniques were also used to identify specific regions of gTBP that may be used to generate synthetic peptides as immunogens for future antibody production. Author Keywords: ChIP, Giardia intestinalis, Homology modelling, Immunoprecipitation, TATA-binding protein, Western Blotting
Characteristics of Models for Representation of Mathematical Structure in Typesetting Applications and the Cognition of Digitally Transcribing Mathematics
The digital typesetting of mathematics can present many challenges to users, especially those of novice to intermediate experience levels. Through a series of experiments, we show that two models used to represent mathematical structure in these typesetting applications, the 1-dimensional structure based model and the 2-dimensional freeform model, cause interference with users' working memory during the process of transcribing mathematical content. This is a notable finding as a connection between working memory and mathematical performance has been established in the literature. Furthermore, we find that elements of these models allow them to handle various types of mathematical notation with different degrees of success. Notably, the 2-dimensional freeform model allows users to insert and manipulate exponents with increased efficiency and reduced cognitive load and working memory interference while the 1-dimensional structure based model allows for handling of the fraction structure with greater efficiency and decreased cognitive load. Author Keywords: mathematical cognition, mathematical software, user experience, working memory
Characterization of Synthetic and Natural Se8 and Related SenSm Compounds by Gas Chromatography-Mass Spectrometry
Elemental selenium has been extensively quantitatively measured in sediments; however, its physical composition is largely unknown, despite it being the dominant selenium species in some reducing environments. Here, for the first time, it is shown that small, cyclic selenium compounds can account for a quantitatively-relevant fraction of the total elemental selenium present. A new method was developed to analyze for cyclooctaselenium (Se8) in both synthetic samples and selenium-impacted sediments. Despite some analytical limitations, this gas chromatography-mass spectrometry (GC-MS) method is the first GC-MS method developed to identify and quantify Se8 in sediments. Once this method was established, it was then applied to more complex systems: first, the identification of compounds in mixed selenium-sulfur melt solutions, and then the determination of SenSm in selenium-impacted sediments. Despite complications arising from pronounced fragmentation in the ion source, assignment of definitive molecular formulae to chromatographically-resolved peaks was possible for five compounds. Developing a fully quantitative method to obtain elemental ratio information can aid in the assignment of molecular formulae to chromatographically-resolved SeS-containing chromatographic peaks. Coupling the existing gas chromatography method to an inductively coupled plasma-mass spectrometer (ICP-MS) system should accomplish this. However, due to a number of complications, this was not completed successfully during the duration of this thesis project. High detection limits for sulfur, retention time discrepancies, and inconsistent injection results between the GC-MS and GC-ICP-MS system led to difficulties in comparing results between both analytical methods. Despite these limitations, GC-ICP-MS remains the most promising method for the identification and quantification of SenSm compounds in synthetic melt mixtures and selenium impacted sediments. Author Keywords: gas chromatography-mass spectrometry, sediments, selenium
Characterization of a Zn(II)2Cys6 transcription factor in Ustilago maydis and its role in pathogenesis
Ustilago maydis (D.C.) Corda is a biotrophic pathogen that secretes effectors to establish and maintain a relationship with its host, Zea mays. In this pathosystem, the molecular function of effectors is well-studied, but the regulation of effector gene expression remains largely unknown. This study characterized Zfp1, a putative U. maydis Zn(II)2Cys6 transcription factor, as a modulator of effector gene expression. The amino acid sequence of Zfp1 indicated the presence of a GAL4-like zinc binuclear cluster as well as a fungal specific transcription factor domain. Nuclear localization was confirmed by tagging Zfp1 with enhanced green fluorescent protein. Deletion of zfp1 resulted in attenuated hyphal growth, reduced infection frequency, an arrest in pathogenic development, and decreased anthocyanin production. This phenotype can be attributed to the altered transcript levels of genes encoding predicted and confirmed U. maydis effectors in the zfp1 deletion strain during pathogenic growth. Complementation of zfp1 deletion strain with tin2, an effector involved in anthocyanin induction, suggested this effector is downstream of Zfp1 and its expression is influenced by this transcription factor during in planta growth. When wild-type zfp1 was ectopically inserted in the zfp1 deletion strain, pathogenesis and virulence were partially restored. This, coupled with zfp1 over-expression strains having a similar phenotype as the deletion strains, suggested Zfp1 may interact with other proteins for full function. These findings show that Zfp1, in conjunction with one or more binding partners, contributes to U. maydis pathogenesis, virulence, and anthocyanin production through the regulation of effector gene expression. Author Keywords: effector, pathogenesis, transcription factor, Ustilago maydis, Zea mays, zinc finger
Characterization of frog virus 3 and its binding partner LITAF
Iridoviruses are large (120-200nm) double stranded DNA viruses that contain an icosahedral capsid. The iridoviridae family is composed of five genera that infect a wide range of poikilothermic vertebrates (Lymphocystivirus, Ranavirus and Megalocyivirus) and invertebrate hosts (Iridovirus, Chloriridovirus). Frog virus 3 (FV3) is a member of the Ranavirus genus, and is commonly used as a model system to study iridoviruses. I was interested in understanding virus-host interaction in FV3. I studied two viral genes, FV3 97R and FV3 75L. Here I demonstrate that 97R localizes to the endoplasmic reticulum (ER) at 24 hours post-transfection. However, at 35 hours post-transfection 97R localizes to the ER but also begins to form concentrated pockets, continuous with the nuclear membrane This study found that 97R possess a unique phenotype and that its localization to the ER is mediated through its C-terminus transmembrane domain. FV3 75L encodes an 84 amino acids protein. I showed that FV3 75L localizes to the early endosomes, while its cellular binding partner, LITAF, localizes to late endosome/lysosome. Interestingly, when FV3 75L and LITAF are co-transfected into cells, LITAF can alter the subcellular localization of FV3 75L to late endosome/lysosomes. A physical interaction between LITAF and FV3 75L was demonstrated through a pull-down assay and that a highly conserved domain found in both proteins may mediate the interaction. LITAF has been proposed to function in protein degradation, but there is still uncertainty on LITAF's specific role. I was interested in further characterizing LITAF and its implications in protein degradation and a neurodegenerative disorder. At least 9 mutations of LITAF are associated with Charcot-Marie-Tooth disease type 1C (CMT1C), which belongs to the group of most common heritable neuromuscular disorders, affecting approximately one in 2500 people. We show that LITAF mutants G112S and W116G mislocalize from the late endosome/lysosome to the mitochondria while the T49M and P135T mutants show partial mislocalization with a portion of the protein present in the late endosome/lysosome and a portion of the protein localized to the mitochondria. Since LITAF is believed to play a role in protein degradation, it is possible that the specific characteristics of CMT1C may occur though impaired degradation of Schwann cell membrane proteins, such as PMP22. I was able to show that when WT LITAF is present, there is a decrease in the PMP22 intracellular levels, which suggest that LITAF plays an important role in protein degradation, and also in other types of CMT. Insight into how mutations in LITAF cause CMT1C may not only help better understand cellular pathways, but also further elucidate the role LITAF's viral homolog FV3 75L during viral infection. Author Keywords: 75L, Charcot-Marie-Tooth, CMTC1, ER, FV3, LITAF
Characterizing the demographic history and prion protein gene variation to infer susceptibility to chronic wasting disease in a naïve population of white-tailed deer (Odocoileus virginianus)
Assessments of the adaptive potential of natural populations are essential for understanding and predicting responses to environmental stressors like climate change and infectious disease. The range of stressors species face in a human-dominated landscape, often have contrasting effects. White-tailed deer (Odocoileus virginianus, deer) are expanding in the northern part of their range following decreasing winter severity and increasing forage availability, caused by climate change. Chronic wasting disease (CWD), a prion disease affecting cervids, is likewise expanding and represents a major threat to deer and other cervids We obtained tissue samples from free-ranging deer across their native range in Ontario, Canada which has yet to detect CWD in wild populations of cervids. High throughput sequencing was used to assess neutral genomic variation and variation in the gene responsible for the protein that misfolds into prions when deer contract CWD, known as the PRNP gene. Neutral variation revealed a high number of rare alleles and no population structure, consistent with an expanding population of deer. Functional genetic variation revealed that the frequencies of variants associated to CWD susceptibility and disease progression were evenly distributed across the landscape and the frequencies were consistent with deer populations not infected with CWD. These findings suggest that an observable shift in PRNP allele frequencies likely coincides with the start of a novel CWD epidemic. Sustained surveillance of genomic and genetic variation can be a useful tool for CWD-free regions where deer are managed for ecological and economic benefits. Author Keywords: Canadian wildlife, population genetics, prion, PRNP, RADseq, ungulate
Chemical characterization of dissolved organic matter in relation with hydrography in the Arctic Ocean
In this thesis, water mass distribution of dissolved organic matter (DOM) characteristics (i.e. molecular weight, fluorescent components, thiols and humic substances concentration) was observed in the Arctic Ocean. For the first time, DOM molecular weight (MW) in Beaufort Sea was assessed using asymmetrical flow field-flow fractionation, as well as the first monitoring of thiols and humic substances (HS) using cathodic stripping voltammetry (CSV) in the Arctic Ocean. Based on fluorescence property, DOM characterization was carried out using parallel factor analysis – excitation-emission matrices. Pacific winter waters in the Canada Basin showed higher MW DOM associated with higher fluorescence intensity. High HS was associated with the Arctic outflow waters in top 300 m of the Canadian Arctic Archipelago. Interestingly, maximum thiol concentration was associated with the subsurface chlorophyll-a maximum at most sites, but not universal along the study area. Comparable distributions of CSV-based HS and humic-like fluorescent components suggest similar sources/ processes in the Arctic Ocean. The findings in this thesis suggested DOM characteristics could be used as fingerprints in tracing water masses in the Arctic Ocean. Author Keywords: Asymmetrical flow field-flow fractionation, Cathodic stripping voltammetry, DOM, Metal-binding ligands, Molecular weight, PARAFAC-EEMs
Chew the Fat
Fatty acid analysis was performed on archaeological bone from various fauna from sites in the Canadian arctic to better understand the preservation of fatty acids and their potential applications to palaeoecological and palaeodietary studies. These data were complemented by analyses of modern bone and soft tissue samples from livestock and harp seals (Pagophilus groenlandicus). Results of modern analyses revealed that in terrestrial species, bone has inherently lower concentrations of most fatty acids relative to other soft tissues (adipose, marrow, and muscle). These analyses suggest that the distribution of fatty acids in bone is unique compared to other tissues, and the types and abundances of fatty acids in bone may be linked to dietary sources of lipids. Of the archaeological samples analyzed, terrestrial species (caribou [Rangifer tarandus]) generally exhibited higher concentrations of saturated fatty acids compared to marine species (ringed seals [Pusa hispida] and polar bears [Ursus maritimus]), whereas marine species had higher concentrations of monounsaturated fatty acids compared to terrestrial species. Results of analyses on both modern and archaeological samples provided insight into the degradation of fatty acids in bone, and the rapid loss of polyunsaturated fatty acids in particular. Because the abundances of fatty acids are likely altered in the burial environment, it is recommended that future analyses incorporate compound specific isotope analysis to focus on applications of fatty acids that are typically in the highest abundance and arguably have undergone the least amount of change, including palmitic (C16:0) and stearic acid (C18:0) Author Keywords: Archaeological Science, Bone Lipids, Fatty Acid Analysis, GC-MS, Lipid Preservation, Palaeoecology
Childhood Precursors of Adult Trait Incompleteness
Previous research has suggested that childhood sensory sensitivity may predict adult obsessive compulsive (OC) behaviours. To date, however, research has not addressed how the separate dimensions – harm avoidance and incompleteness - may influence this relationship or why it exists. The current study used a retrospective design to test a) if sensory sensitivity in childhood predicts trait incompleteness in adulthood, as well as b) if emotion regulation variables mediate this relationship. Questionnaires pertaining to OC dimensions and childhood anxieties were completed independently by 172 undergraduate participants and their primary childhood caregiver. Results showed a linear relationship between sensory sensitivity in childhood and incompleteness in adults. Emotion regulation variables failed to mediate this relationship, although a trend for mediation was present. Additionally, exploratory analysis found perfectionism in childhood to be a predictor of trait incompleteness but not harm avoidance, whereas physical anxieties predicted harm avoidance and not incompleteness. Results are discussed in the context of clinical and theoretical implications. Author Keywords: Distress Tolerance, Harm Avoidance, Incompleteness, Obsessive Compulsive Disorder, Sensory Sensitivity, Symmetry
Childhood diet and feeding practices at Apollonia
This study analyses deciduous dental pathology and stable carbon and nitrogen isotopes to investigate the relationship between dietary composition, feeding practices, and oral health in a subadult skeletal sample from the Greek colonial site of Apollonia Pontica, Bulgaria (5th to 3rd century BC). Stable isotope analysis of 74 bone collagen samples indicates that weaning began between the ages of 6 months and 1 year, and was complete by the age of 4. The stable isotope data are consistent with a diet of primarily terrestrial C3 resources. The deciduous dentitions of 85 individuals aged between 8.5 months and 10.5 years were examined for evidence of a number of pathological conditions. The presence of dental caries, calculus, occlusal tooth wear and an abscess indicate that foods introduced early in life affected the oral health of these individuals. Overall, the deciduous dental data correlate well with the stable isotope data and ancient textual sources regarding infant and childhood dietary composition and feeding practices. Author Keywords: breastfeeding, deciduous dentition, dental pathology, stable carbon and nitrogen isotopes, weaning

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Format: 2024/02/21