Role Of Peptidoglycan Modifications In Predation By Bdellovibrio Bacteriovorus 109j, Diane C. Szmiett

Electronic Thesis and Dissertation Repository

Bdellovibrio bacteriovorus is a prey dependent bacterium that traverses the outer membrane and peptidoglycan, and enters the periplasmic space of gram negative bacteria, from where it utilizes the cytoplasmic contents for growth. Soluble lytic transglycosylases could be responsible for creating the entry pore during predation. Bacteria can modify their peptidoglycan by the addition of acetate to the C-6 hydroxyl group of the N-acetylmuramic acid residues, which renders the peptidoglycan insensitive to cleavage by lytic transglycosylases. It was hypothesized that the degree of peptidoglycan O-acetylation of the prey cell affects predation efficiency. In this study it was shown that; …

Production And Applications Of Formaldehyde-Free Phenolic Resins Using 5-Hydroxymethylfurfural Derived From Glucose In-Situ, Yongsheng Zhang

Electronic Thesis and Dissertation Repository

The phenol-formaldehyde (PF) resin manufacturing industry is facing a growing challenge with respect to concerns over human health, due to the use of carcinogenic formaldehyde and sustainability due to the use of petroleum-based phenol in PF resin manufacture. Glucose and its derivative, 5-hydroxymethylfurfural (5-HMF), have proven to be potential substitutes for formaldehyde in the synthesis of phenolic novolac resins.

This thesis investigated a number of glucose and 5-HMF resin systems including the curing of phenol-glucose novolac resin (PG) with a bis-phenol-A type epoxy. The curing process was modeled according to the Sestak-Berggren equation (S, B) using Málek methods. This was …

Response Of Soil Nitrification To The Veterinary Pharmaceuticals Monensin, Ivermectin And Zinc Bacitracin, Magda A. Konopka

Electronic Thesis and Dissertation Repository

Pharmaceutical residues can reach agricultural land through amendment with animal or human waste. Since 2010, a series of replicated plots received annual applications of ivermectin, monensin and zinc bacitracin, either singly or in a mixture, at 0.1 mg/kg and 10 mg/kg concentrations. I collected soil samples before and after the fourth annual application of pharmaceuticals and assayed them for functional changes and amoA gene abundance, a gene needed for ammonia oxidation. In 2013, I exposed the soils to 100 mg/kg in a laboratory experiment which resulted in acceleration of nitrification. Under 10 mg/kg treatments in the field the abundance of …

The Role Of Islands In The Migration Of Bats Across Lake Erie And Lake Ontario: Lasiurus Borealis, Lasiurus Cinereus And Perimyotis Subflavus, Toby J. Thorne

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Large bodies of water, such as the Great Lakes in North America, are potential barriers to migrating bats. Islands may be used as stopover sites by bats crossing the Great Lakes as stopover sites, or included in crossing routes to reduce the distance flown over open water. I predicted that stopover use by migrating bats would result in higher activity on Great Lakes islands during late-season migration than at mainland sites. I monitored acoustic activity at 11 sites across four locations in Ontario: Amherst and Pelee islands, Kingston and Pinery Provincial Park during the summer (June and July), and during …

Characterizing Persistent Developmental Dyscalculia: A Cognitive Neuroscience Approach, Stephanie D. Bugden

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Developmental dyscalculia (DD) is a specific learning disorder of calculation abilities. In the present thesis I report a series behavioural and functional neuroimaging studies to further elucidate the core numerical deficits underlying DD. I recruited a sample of children with DD who demonstrated persistent impairments in arithmetic. In Chapter 2, to validate the selection criteria, I compared the performance of children with and without persistent DD on a test of numerical magnitude processing. The data showed that only children with persistent DD presented with deficits in numerical magnitude processing, while those with inconsistent DD perform at the level of age-matched …

Inheritance Of Virulence In The Root Rot Pathogen Phytophthora Sojae, Sirjana Devi Shrestha

Electronic Thesis and Dissertation Repository

The oomycete Phytophthora sojae causes stem and root rot of soybean plants. The interaction of pathogen avirulence (Avr) and host resistance (R)-genes determine the disease outcome. The Avr3a mRNA transcript level is variable among P. sojae strains and determines virulence towards the R-gene Rps3a. To study the inheritance of virulence, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 and P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F₅ generation. However, test-crosses of F₁ (Avr3a^ACR10/Avr3a …

Role Of Integrin-Linked Kinase In Epidermal Integrity And Barrier Function, Samar Sayedyahossein

Electronic Thesis and Dissertation Repository

Integrin-linked kinase (ILK) is a β integrin adaptor protein that translates extracellular stimuli to intracellular signaling events. ILK plays a role in actin cytoskeleton dynamics and cell adhesion. The structure and function of the epidermis is highly dependent on cell-cell adhesion and cell-basement membrane interactions. The mechanisms whereby ILK contributes to epidermal integrity are poorly understood. Using a mouse model of epidermis-restricted Ilk gene inactivation, I observed that ILK loss causes abnormal morphology and presence of intra-epidermal and epidermal-dermal microblisters in embryos as early as E17.5. ILK-deficient epidermis is also characterized by abnormal localization or/and absence of adherens junctions, tight …

A Comparison Of Community Composition Analyses For The Assessment Of Responses To Wood-Ash Soil Amendment By Free-Living Nematodes, Paul B.L. George

Electronic Thesis and Dissertation Repository

Land-use changes can have far-reaching consequences for resident communities and ecosystem functioning. Developing appropriate assessment methods to observe and quantify this change is an important application of community ecology. Here I compare four methods of community assessment for free-living soil nematodes under forest harvesting disturbance and wood ash application. Neither morphological assessment (richness, abundance, diversity) nor molecular assessment (morpho-richness using T-RFLP) was responsive to experimental treatments. Trait-based approaches (Maturity Index (MI) and Body Size Spectra (BSS)) were more sensitive to forest harvest and wood-ash amendment treatments. The efficacy of these methods was also qualitatively compared. Of all methods, the BSS …

Neuromuscular Adaptations To Voluntary Contraction Following Postactivation Potentiation, Cameron B. Smith

Electronic Thesis and Dissertation Repository

Muscle contractile properties are history-dependent, and following a conditioning contraction, muscle tissue may be fatigued (slower, weaker) or potentiated (faster, stronger). Postactivation potentiation of evoked contractions, such as the electrically stimulated twitch, has been thoroughly studied. However, the effects of potentiation on voluntary contraction are not well understood, and prior study is largely equivocal.

The following studies propose to determine the effects of potentiation during 1) submaximal contractions at different muscle lengths 2) ballistic contractions following tetanic and voluntary conditioning, and 3) motor evoked potentials following tetanic and voluntary conditioning contractions. Evoked twitch potentiation was assessed with all of the …

Hcpe, A Potential Immuno-Modulatory Protein From Helicobacter Pylori That Is Dependent On The Disulfide Bond Protein Dsbhp, Jeff Lester

Electronic Thesis and Dissertation Repository

H. pylori is a human gastric pathogen that colonizes ~ 50% of the world’s population. It can cause gastritis, gastric or duodenal ulcers and also gastric cancer. H. pylori produces Helicobacter cysteine rich protein HcpE, a secreted protein which may play a role in virulence. In this study we show that HcpE is secreted in the culture supernatant both as a soluble protein and in association with outer membrane vesicles, and may play a role in the modulation of H. pylori inflammatory responses. We identified that Dsb_HP is necessary for HcpE production and secretion in H. pylori, and …

Effect Of Photoperiod On Redox Regulation Of Phenotypic Plasticity And Cellular Growth In Chlorella Vulgaris, Lauren E. Hollis

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Photoautotrophs are predisposed to maintain a balance between light energy absorption with the capacity to consume this energy through metabolism. An imbalance in energy flow may be a consequence of increased light intensity and is sensed as modulation of excitation pressure (EP). Chlorella vulgaris acclimated to continuous high EP exhibits a yellow-green phenotype characterized by reduced chlorophyll content and high chlorophyll a/b ratio with reduced light-harvesting complex abundance relative to the dark green phenotype of low EP-acclimated cultures. Previous studies on acclimation to EP in green algae have been conducted under constant growth light. To determine the role of EP …

Thylakoid Phosphorylation And Cell Morphology In The Antarctic Psychrophile, Chlamydomonas Sp. Uwo241, Beth Szyszka-Mroz

Electronic Thesis and Dissertation Repository

The unicellular green microalga, Chlamydomonas sp. UWO 241, was isolated from Lake Bonney, Antarctica. A unique characteristic of this algal strain is its inability to undergo state transitions combined with an altered thylakoid protein phosphorylation profile, which suggests the absence of LHCII phosphorylation, and preferential phosphorylation of a set of novel proteins. Examination of the unique phosphoproteins revealed that they are associated with a large pigment-protein supercomplex, which contains components of both photosystem I and the cytochrome b₆/f complex and likely functions in cyclic electron flow (CEF).

The absence of phosphorylation of LHCII proteins, associated with state …

Validation Of The Multi-Segment Foot Model With Bi-Planar Fluoroscopy, Aïda Valevicius

Electronic Thesis and Dissertation Repository

A multi-segment foot model (MSFM) is a useful tool for measuring foot joint kinematics although soft-tissue artefact is often present. Quantifying this error is needed to evaluate the accuracy of this model. This study validated the MSFM against bi-planar radiostereometric analysis (RSA) fluoroscopy. Heel-strike, mid-stance, and toe-off events during the stance phase were compared between motion capture and fluoroscopy. Rise/drop of the medial longitudinal arch showed a significant difference (p < 0.05) during toe-off, but no significant difference during heel-strike or mid-stance. Hindfoot supination/pronation and internal/external rotation, and forefoot supination/pronation motions showed no significant difference between the two techniques. The lack of significant difference will allow the MSFM to be used as a sufficiently accurate technique for measuring foot joint motions.

Low Temperature Tolerance Of Adult Drosophila Suzukii (Diptera: Drosophilidae), Ruth Jakobs

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The crop pest Drosophila suzukii, native to Southeast Asia, has been found in Ontario since 2010. This raises concern with respect to it establishing permanent populations, however, little is known about its cold tolerance. I investigated the low-temperature tolerance, including phenotypic plasticity, of D. suzukii. While acclimation increased cold tolerance, there was no evidence of short-term cold-hardening. Chill coma occurs at -1.2 °C, which will limit winter activity. Cold shock decreased the reproductive output of females, but this negative effect may be mitigated by re-mating. Drosophila suzukii is chill-susceptible and 80 % of the flies die after exposure …

Novel Mechanisms In The Sorting Of Proglucagon To The Secretory Granules Of The Regulated Secretory Pathway, Leonard M. Guizzetti

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The prohormone proglucagon encodes for multiple peptide hormones, including glucagon, glucagon-like peptide-1 (GLP-1), and GLP-2, produced through tissue-specific processing by prohormone convertase (PC) 1/3 and PC2. In alpha cells, PC2 yields glucagon, the major counter-regulatory hormone to insulin, which together, control glucose homeostasis. In contrast, GLP-1 and GLP2 are mainly produced in intestinal L-cells by PC1/3. GLP-1 stimulates insulin secretion following a meal, and therefore has opposing function to glucagon regulating glucose homeostasis; in contrast, GLP-2 enhances gut nutrient absorption. Efficient sorting of proglucagon to secretory granules is required for nutrient-regulated secretion. The aim of this thesis is to discover …

Arabidopsis Chromatin Remodeler Brahma: Its Functional Interplay With Polycomb Proteins And The Ref6 Histone Demethylase, Chenlong Li

Electronic Thesis and Dissertation Repository

BRAHMA (BRM) is a SWI/SNF-type chromatin remodeling ATPase that plays an important role in regulation of gene expression. Tri-methylation of lysine 27 on histone H3 (H3K27me3) is a histone modification that is associated with transcriptionally repressed genes and catalyzed by Polycomb Group (PcG) proteins. BRM has been proposed to antagonize the function of PcG proteins but the underlying molecular mechanism is unclear. To understand how BRM regulates the function of PcG proteins during plant development, a genome-wide analysis of H3K27me3 in brm mutant was performed using chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq). Loss of BRM leads to increased …

Application Of Otolith Increment Analysis To The Study Of Maturation Timing In Female Kokanee Salmon, Yelin Xu

Electronic Thesis and Dissertation Repository

I investigated the influence of growth history on the expression of female reproductive tactics in kokanee salmon (Oncorhynchus nerka) from Meadow Creek Spawning Channel, British Columbia, Canada. Female kokanee either arrive at the spawning area with red nuptial coloration, or less commonly, sexually immature with silver coloration. Silver- and red-arriving females may reflect different reproductive strategies in the population. I used otolith increment measurements to determine fish growth. In contrast to earlier studies, silver- and red-arriving females in 2013 did not differ in age at maturity (mostly were age 3+) or size at maturity (length from eye to …

Pathway-Specific Signaling And Its Impact On Fertility: A Focus On The Kisspeptin Receptor, Kiss1r, Maryse R. Ahow

Electronic Thesis and Dissertation Repository

Hypothalamic GnRH-releasing hormone (Gnrh) is the master regulator of the neuroendocrine reproductive axis and its secretion is regulated by many Gnrh neuronal-based signaling systems. Among these are Gα_q/11-coupled receptors and their ligands.In most cases examined to date, activation of these receptors lead to Gnrh neuronal membrane depolarization and Gnrh secretion. The most potent trigger of Gnrh secretion is kisspeptin (Kp), a ligand for the Kiss1r Gα_q/11-coupled receptor. Studies have shown that Kiss1r signaling is essential for attaining and maintaining fertility. We recently demonstrated that in addition to signaling via Gα_q/11, Kiss1r also couples to …

Protein Body Biogenesis And Utility In Recombinant Protein Production In Nicotiana Benthamiana, Reza Saberianfar

Electronic Thesis and Dissertation Repository

Protein bodies (PBs) are endoplasmic reticulum (ER) derived organelles found in seeds whose function is to accumulate seed storage proteins. It was shown that PB formation is not limited to seeds, and green fluorescent protein (GFP) fused to either elastin-like polypeptide (ELP), hydrophobin-I (HFBI) or Zera^® fusion tags induces PBs in leaves of Nicotiana benthamiana. The mechanism by which fusion tags induce PBs is not well understood. To address how PBs form and develop in plant leaves, I studied the factors involved in their formation including recombinant protein concentration, effect of the fusion tags, PB sequestration patterns and …

Characterization Of Aldh Positive Cells In The Human Fetal Pancreas, Amanda Oakie

Electronic Thesis and Dissertation Repository

Aldehyde dehydrogenase (ALDH) is an enzyme that regulates differentiation during pancreatic development, but determining the role of ALDH during endocrine differentiation is not well defined. The purpose of this study was to characterize transcription factors (TFs) and putative stem cell surface markers in ALDHhi or ALDHlo populations of the human fetal pancreas (18-22 weeks of age), and examine their expansion and differentiation in vitro and in vivo. Cell surface marker analysis demonstrated high co-localization of the stem cell marker CD133 with ALDHhi cells. ALDHhi cells also contained higher expression of islet- associated TFs and insulin. Expansion of sorted ALDHhi cells …

The Role Of The Ku70 Vwa Domain In The Response To Dna Double Strand Breaks, Victoria L. Fell

Electronic Thesis and Dissertation Repository

Ku is an abundant, highly conserved DNA binding protein found in both prokaryotes and eukaryotes that plays essential roles in the maintenance of genome integrity. In eukaryotes, Ku is a heterodimer comprised of two subunits, Ku70 and Ku80, and is best characterized for its central role as the initial DNA end binding factor in the “classical” non-homologous end joining (C-NHEJ) pathway, the main DNA double-strand break (DSB) repair pathway in mammals. At the break, Ku directly and indirectly interacts with several C-NHEJ factors and processing enzymes, serving as the scaffold for the entire DNA repair complex. In this work we …

Acquisition And Reconstruction Techniques For Fat Quantification Using Magnetic Resonance Imaging, Abraam S. Soliman

Electronic Thesis and Dissertation Repository

Quantifying the tissue fat concentration is important for several diseases in various organs including liver, heart, skeletal muscle and kidney. Uniquely, MRI can separate the signal from water and fat in-vivo, rendering it the most suitable imaging modality for non-invasive fat quantification. Chemical-shift-encoded MRI is commonly used for quantitative fat measurement due to its unique ability to generate a separate image for water and fat. The tissue fat concentration can be consequently estimated from the two images. However, several confounding factors can hinder the water/fat separation process, leading to incorrect estimation of fat concentration.

The inhomogeneities of the main …

Changing The Substrate Specificity Of Arogenate Dehydratases (Adts) From Arabidopsis Thaliana., Megan Es Smith-Uffen

Electronic Thesis and Dissertation Repository

Phenylalanine (Phe), an essential aromatic amino acid, serves as a precursor for protein synthesis and a variety of secondary metabolites in plants. Two pathways are known for Phe biosynthesis. In the first, prephenate dehydratases (PDTs) convert prephenate to phenylpyruvate, which is transaminated to Phe. In the second, prephenate is transaminated to arogenate, which is converted to Phe by arogenate dehydratases (ADTs). ADTs and PDTs are structurally very similar, as are their substrates. Six ADTs (ADT1-ADT6) have been identified in Arabidopsis thaliana. ADT1 and ADT2 can recognize both prephenate and arogenate as substrates whereas ADT3-ADT6 are solely arogenate-accepting. Twenty ADT domain-swapping …

Social Immunity And The Expression Of Immune-Related Genes In The Eastern Subterranean Termite, Qi Gao

Electronic Thesis and Dissertation Repository

Individuals living within social groups may benefit from the efficiencies of division of labour, but on the other hand render themselves vulnerable to socially transmitted disease. This cost to social living should promote cooperative barriers to disease transmission, especially in eusocial taxa where spatial and genetic proximity to nestmates are characteristically pronounced. Though studies of the immunity at multiple levels in social species are becoming more common, little is known about how their sociality is deployed to resist contagion. By exposing the Eastern subterranean termite Reticulitermes flavipes to entomopathogenic fungi, I studied their immune responses at multiple levels. At the …

The Role Of Superantigens During Staphylococcus Aureus Nasal Colonization And Infection, Stacey Xu

Electronic Thesis and Dissertation Repository

Superantigens (SAgs) are potent toxins produced by bacteria such as Staphylococcus aureus that function to overactivate T cells resulting in massive cytokine production and immune activation. Despite decades of research on the structure and function of these proteins, as well as their role in severe diseases such as toxic shock syndrome, the question as to why strains of S. aureus produce SAgs and the role that they play in the life cycle of these bacteria remains unanswered. The contribution of SAgs towards pathogenicity and bacterial survival in vivo were assessed using isogenic SAg deletion knockouts in conjunction with SAg-sensitive humanized …

Regulation Of Igfbp-1 Phosphorylation In Hypoxia Via Mtor Signaling, Ian Damerill

Electronic Thesis and Dissertation Repository

In this study, we provide novel evidence for a role of fetal liver mTOR signaling in regulating IGF-I bioavailability by modulating IGFBP-1 phosphorylation due to hypoxia – a key factor in the development of reduced fetal growth in utero. We utilized HepG2 cells in vitro and demonstrated a link between mTOR inhibition and hypoxia-induced IGFBP-1 phosphorylation. Using a biological assay for IGF-I receptor autophosphorylation, we demonstrated a functional significance for hypoxia-induced IGFBP-1 phosphorylation in reducing IGF-I bioactivity in vitro. Further, we have implicated a mechanistic link to increased CK2 activity within this regulation. We demonstrate that mTOR inhibition …

Post-Translational Control Of Retinoblastoma Protein Phosphorylation, Paul M. Stafford

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The retinoblastoma tumor suppressor protein (pRB) functions through multiple mechanisms to serve as a tumor suppressor. pRB has been well characterized to be inactivated through phosphorylation by CDKs. pRB dephosphorylation and activation is a much less characterized aspect of pRB function. In this thesis, I detail work to study the post translational control of pRB phosphorylation. Here I present work detailing efforts to generate a gene targeted mouse which disrupts PP1 binding to the C-terminus of pRB, allowing for detailed study of the mechanisms of pRB dephosphorylation. This work also details an examination of acetylation in the C-terminus of pRB, …

A Rhog/Ilk/Elmo2/Rac1 Pathways Modulates Microtubule Stability, Bradley C. Jackson

Electronic Thesis and Dissertation Repository

Integrin-linked kinase (ILK) is a widely expressed scaffold protein important for cell adhesion, migration, and phagocytosis. ILK and its interacting partner, engulfment and cell motility protein 2 (ELMO2), may also modulate microtubule dynamics. Here, I show that sustained microtubule growth and acetylated tubulin levels are reduced in the absence of ILK and, conversely, that exogenous ILK exerts a stabilizing effect on microtubules. Further, exogenous ELMO2, ras homology growth related protein (RHOG) and ras-related C3 botulinum toxin substrate 1 (RAC1) stabilize microtubules in an ILK-dependent manner. Potential downstream effectors of the effect of ILK on microtubule stability include glycogen synthase kinase- …

The Role Of The Rna-Binding Protein Rho Guanine Nucleotide Exchange Factor In The Cellular Stress Response, Kevin Wh Cheung

Electronic Thesis and Dissertation Repository

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease for which the pathological mechanism is heterogeneous and a cure has been elusive. Recent developments have linked specific proteins found in pathological neuronal cytoplasmic inclusions (NCIs) of ALS motor neurons to familial variants of the disease. These proteins, including TAR DNA-binding protein of 43 kDa (TDP-43), fused in sarcoma/translocated in liposarcoma (FUS), and Rho guanine nucleotide exchange factor (RGNEF) share the common characteristic of being RNA-binding proteins that colocalize within NCIs. RGNEF is unique however in also possessing RhoA activation capacity, suggesting a role in the cell stress response. My thesis …

Pregestational Diabetes And Congenital Heart Defects: Role Of Reactive Oxygen Species, Hoda Moazzen

Electronic Thesis and Dissertation Repository

Women with pregestational diabetes are at a high risk of having babies with congenital heart defects (CHDs). The mechanisms of CHD induction in the offspring of women with pregestational diabetes remain elusive. With an ever-increasing rate of diabetes in young adults, there is a pressing need to understand the underlying mechanisms and initiate effective preventative strategies. The aim of this thesis is to study the role of reactive oxygen species (ROS) signalling in heart morphogenesis, unravel molecular mechanisms of CHDs induced by pregestational diabetes and provide new insights on potential therapeutic strategies. To this end, a mouse model of pregestational …