• The Molecular Consequences of CK2-mediated Phosphorylation of the TGA2 Transcription Factor within Systemic Acquired Resistance of Arabidopsis thaliana

      Bosak, Jan; Centre for Biotechnology (Brock University, 2014-08-05)
      During infection, the model plant Arabidopsis thaliana is capable of activating long lasting defence responses both in tissue directly affected by the pathogen and in more distal tissue. Systemic acquired resistance (SAR) is a type of systemic defence response deployed against biotrophic pathogens resulting in altered plant gene expression and production of antimicrobial compounds. One such gene involved in plant defence is called pathogenesis-related 1 (PR1) and is under the control of several protein regulators. TGA II-clade transcription factors (namely TGA2) repress PR1 activity prior to infection by forming large oligomeric complexes effectively blocking gene transcription. After pathogen detection, these complexes are dispersed by a mechanism unknown until now and free TGA molecules interact with the non-expressor of pathogenesis-related gene 1 (NPR1) protein forming an activating complex enabling PR1 transcription. This study elucidates the TGA2 dissociation mechanism by introducing protein kinase CK2 into this process. This enzyme efficiently phosphorylates TGA2 resulting in two crucial events. Firstly, the DNA-binding ability of this transcription factor is completely abolished explaining how the large TGA2 complexes are quickly evicted from the PR1 promoter. Secondly, a portion of TGA2 molecules dissociate from the complexes after phosphorylation which likely makes them available for the formation of the TGA2-NPR1 activating complex. We also show that phosphorylation of a multiserine motif found within TGA2’s N terminus is responsible for the change of affinity to DNA, while modification of a single threonine in the leucine zipper domain seems to be responsible for deoligomerization. Despite the substantial changes caused by phosphorylation, TGA2 is still capable of interacting with NPR1 and these proteins together form a complex on DNA promoting PR1 transcription. Therefore, we propose a change in the current model of how PR1 is regulated by adding CK2 which targets TGA2 displacing it’s complexes from the promoter and providing solitary TGA2 molecules for assembly of the activating complex. Amino acid sequences of regions targeted by CK2 in Arabidopsis TGA2 are similar to those found in TGA2 homologs in rice and tobacco. Therefore, the molecular mechanism that we have identified may be conserved among various plants, including important crop species, adding to the significance of our findings.
    • Molecular ecology and social evolution of the eastern carpenter bee, Xylocopa virginica

      Vickruck, Jessica L; Department of Biological Sciences
      Bees are extremely valuable models in both ecology and evolutionary biology. Their link to agriculture and sensitivity to climate change make them an excellent group to examine how anthropogenic disturbance can affect how genes flow through populations. In addition, many bees demonstrate behavioural flexibility, making certain species valuable models with which to study the evolution of social groups. This thesis studies the molecular ecology and social evolution of one such bee, the eastern carpenter bee, Xylocopa virginica. As a generalist native pollinator that nests almost exclusively in milled lumber, anthropogenic disturbance and climate change have the power to drastically alter how genes flow through eastern carpenter bee populations. In addition, X. virginica is facultatively social and is an excellent organism to examine how species evolve from solitary to group living. Across their range of eastern North America, X. virginica appears to be structured into three main subpopulations: a northern group, a western group and a core group. Population genetic analyses suggest that the northern and potentially the western group represent recent range expansions. Climate data also suggest that summer and winter temperatures describe a significant amount of the genetic differentiation seen across their range. Taken together, this suggests that climate warming may have allowed eastern carpenter bees to expand their range northward. Despite nesting predominantly in disturbed areas, eastern carpenter bees have adapted to newly available habitat and appear to be thriving. This is in marked contrast to many other bee species, particularly in the genus Bombus, who appear unable to shift their ranges along with climate change. Facultatively social organisms are interesting species to study the evolution of social groups, and the remaining chapters address questions of sociality in X. virginica. I used observation nests and genetic relatedness to examined how females behave towards one another in the spring prior to the establishment of dominance hierarchies in social nests. In spring, females directed fewer aggressive behaviours and more cooperative behaviours towards familiar rather than related individuals, indicating that females use nestmate recognition rather that kin recognition when interacting with conspecifics. Overwintering groups often contain both related and unrelated individuals, indicating that many bees interacting with one another in the fall prior to overwintering may be unrelated, emphasizing the importance of recognizing nestmates. Within social carpenter bee nests three different types of female have been described: primary, secondary and tertiary. Primary females are the dominant foragers and egg layers in the nest while secondary and tertiary females appear to join a reproductive queue behind the primary. To understand the nature and flexibility of this reproductive queue I performed removal experiments across three different years. This study showed that secondary females always assumed the role of replacement primary, while tertiary females rarely opted to forage and reproduce even if they were the only female in the nest. Removal experiments demonstrated that social groups in X. virginica are complex and comprise two different reproductive strategies (breed in the current year or delay reproduction) as well as form dominance hierarchies among primary and secondary females. Several tertiary females were able to become primary or solitary females in their second summer, providing evidence for how each type of female may have evolved in social nests. Finally, I examined how competition influences the evolution and maintenance of social groups in eastern carpenter bees. In conditions of high population density significantly more social nests were present in the population, indicating that competition for limiting nesting resources drives individuals together into social groups. Within social groups relatedness was low, and siblings actually dispersed away from one another to other nests in the population, reducing competition among kin. Eastern carpenter bees appear to demonstrate an interesting evolutionary route to sociality, where very high levels of competition among kin lead to dispersal, while limited nesting substrate forces individuals back into unrelated social groups. While predicted by kin selection, social groups of this nature are previously undescribed in the Hymenoptera, and further study of eastern carpenter bees can provide novel insights into alternate routes to sociality.
    • Molybdenum (IV) imido silylamido and hydride complexes : stoichiometric and catalytic reactivity, mechanistic aspects of hydrosilation reactions

      Khalimon, Andrey Y.; Department of Chemistry (Brock University, 2011-05-17)
      This thesis describes the synthesis, structural studies, and stoichiometric and catalytic reactivity of novel Mo(IV) imido silylamide (R'N)Mo(R2)(173_RIN-SiR32-H)(PMe3)n (1: Rl = tBu, Ar', Ar; R2 = Cl; R32 = Me2, MePh, MeCl, Ph2, HPh; n = 2; 2: R' = Ar, R2 = SiH2Ph, n = 1) and hydride complexes (ArN)Mo(H)(R)(PMe3)3 (R = Cl (3), SiH2Ph (4». Compounds of type 1 were generated from (R'N)Mo(PMe3)n(L) (5: R' = tBu, Ar', Ar; L = PMe3, r/- C2H4) and chlorohydrosilanes by the imido/silane coupling approach, recently discovered in our group. The mechanism of the reaction of 5 with HSiCh to give (ArN)MoClz(PMe3)3 (8) was studied by VT NMR, which revealed the intermediacy of (ArN)MCh(172 -ArN=SiHCl)(PMe3)z (9). The imido/silyl coupling methodology was transferred to the reactions of 5 with chlorine-free hydrosilanes. This approach allowed for the isolation of a novel ,B-agostic compound (ArN)Mo(SiHzPh)(173 -NAr-SiHPhH)(PMe3) (10). The latter was found to be active in a variety of hydrosilation processes, including the rare monoaddition of PhSiH3 to benzonitrile. Stoichiometric reactions of 11 with unsaturated compounds appear to proceed via the silanimine intermediate (ArN)M(17z-ArN=SiHPh)(PMe3) (12) and, in the case of olefins and nitriles, give products of Si-C coupling, such as (ArN)Mo(R)(173 -NAr-SiHPh-CH=CHR')(PMe3) (13: R = Et, R' = H; 14: R = H, R' = Ph) and (ArN)Mo(172-NAr-SiHPh-CHR=N)(PMe3) (15). Compound 13 was also subjected to catalysis showing much improved activity in the hydrosilation of carbonyls and alkenes. Hydride complexes 3 and 4 were prepared starting from (ArN)MoCh(PMe3)3 (8). Both hydride species catalyze a diversity of hydrosilation processes that proceed via initial substrate activation but not silane addition. The proposed mechanism is supported by stoichiometric reactions of 3 and 4, kinetic NMR studies, and DFf calculations for the hydrosilation of benzaldehyde and acetone mediated by 4.
    • A Multi-Scale Molecular Dynamic Approach to the Study of the Outer Membrane of the Bacteria Psudomonas Aeruginosa PA01 and the Biocide Chlorhexidine

      Van Oosten, Brad; Department of Physics
      The introductory chapters of this thesis contains an explanation to the methods and basic theory of the molecular dynamics approach. Together with the appendix section, in which a step by step tutorial how to set up and run basic simulations using the gromacs software is presented, this thesis can serve as an introductory aid in performing molecular dynamics simulations. In the research portion of this thesis, I provide several uses for the molecular dynamics approach applied to the biocide chlorhexidine as well as the study of membranes, including a mimic of the bacteria membrane of Pseudomonas Aeruginosa PA01. The motivation for this research was previous work done in our lab which determined that chlorhexidine has a high affinity for DMPC and found the depth at which it resides in a model DMPC membrane. From this information, an all-atom representation of chlorhexidine was made, which was proven to reproduce the experimental results. While we learned much about chlorhexidine in a model DMPC membrane, this study lacked the destruction of the membrane as well as the study of chlorhexidine in a biologically relevant membrane. For these reasons coarse grained versions of the all-atom chlorhexidine models as well as a new lipopolysaccharide molecule was created. With the coarse grained model of chlorhexidine and the ability to create a bacterial membrane mimic, the study of chlorhexidine and other antibacterial agents can be further studied.
    • A Narrative Study of Patient Encounter Accounts of Physicians, Nurses, and Medical Receptionists after Two Decades of a Paradigm of Patient-Centered Care

      Akseer, Riaz; Applied Health Sciences Program (Brock University, 2014-12-02)
      Despite recent well-known advancements in patient care in the medical fields, such as patient-centeredness and evidence-based medicine and practice, there is rather less known about their effects on the particulars of clinician-patient encounters. The emphasis in clinical encounters remains mostly on treatment and diagnosis and less on communicative competency or engagement for medical professionals. The purpose of this narrative study was to explore interactive competencies in diagnostic and therapeutic encounters and intake protocols within the context of the physicians’, nurses’, and medical receptionists’ perspectives and experiences. Literature on narrative medicine, phenomenology and medicine, therapeutic relationships, cultural and communication competency, and non-Western perspectives on human communication provided the guiding theoretical frameworks for the study. Three data sets including 13 participant interviews (5 physicians, 4 nurses, and 4 medical receptionists), policy documents (physicians, nurses, and medical receptionists) and a website (Communication and Cultural Competency) were used. The researcher then engaged in triangulated analyses, including N-Vivo, manifest and latent, Mishler’s (1984, 1995) narrative elements and Charon’s (2005, 2006a, 2006b, 2013) narrative themes, in recursive, overlapping, comparative and intersected analysis strategies. A common factor affecting physicians’ relationships with their clients was limitation of time, including limited time (a) to listen, (b) to come up with a proper diagnosis, and (c) to engage in decision making in critical conditions and limited time for patients’ visits. For almost all nurse participants in the study establishing therapeutic relationships meant being compassionate and empathetic. The goals of intake protocols for the medical receptionists were about being empathetic to patients, being an attentive listener, developing rapport, and being conventionally polite to patients. Participants with the least iv amount of training and preparation (medical receptionists) appeared to be more committed to working narratively in connecting with patients and establishing human relationships as well as in listening to patients’ stories and providing support to narrow down the reason for their visit. The diagnostic and intake “success stories” regarding patient clinical encounters for other study participants were focused on a timely securing of patient information, with some acknowledgement of rapport and emapathy. Patient-centeredness emerged as a discourse practice, with ambiguous or nebulous enactment of its premises in most clinical settings.
    • Negotiating a Gendered Neo-Calvinist Pillar: Immigrant Loss, Transformation, and Lifelong Learning

      VanderVliet, Catharina F.; Department of Graduate and Undergraduate Studies in Education
      Employing a critical feminist perspective, I conducted a sociocultural analysis of the lifelong learning of Dutch neo-Calvinist women who immigrated to Canada shortly after World War II. The purpose of the research was a critique of the institutional ruling relations (schooling, religion, family, workplace) that shaped and influenced the trajectory of these women’s lifelong learning. More specifically, the inquiry included an interrogation of their Canadian schooling experience, in the context of an immigrant family life, their pillarized Dutch culture, and Calvinist religiosity. In choosing a life history methodology, the scope of the research broadened where one’s life story was juxtaposed to a theory of context. Applying this methodology, I critically analyzed structures, operations, and contestations of power in lifelong learning institutions through an exploration of the multiple contexts that shaped the lives of immigrant women. It is within that relationship that the critical feminist was possible. The life histories were not a description of the mainstream but rather were positioned to dialectically interrogate the meaning and significance of the past as it influenced the present and future. Applying a dialectic method to the participants’ life histories, 7 tensions were raised that made visible ruling relations relevant to the participants’ everyday experiences and brought awareness to the underlying contextual and ideological assumptions related to their trajectory of lifelong learning. Employing a critical feminist perspective, I examined how 3 neo-Calvinist immigrant women interpreted and negotiated the ambiguity created by cultural contradictions experienced in a Canadian context. As a researcher who herself has been shaped by this specific immigrant experience, a key attribute of life history methodology was its capacity for the researcher self to be visible in the research.
    • Neural and Behavioural Consequences of Chronic Inflammation following Spinal Cord Injury

      Allison, David; Applied Health Sciences Program
      This thesis investigated the influence of chronic inflammation on several neural/behavioural disorders following spinal cord injury (SCI) including depression, cognitive impairment, neuropathic pain, and somatic nerve deficits. Ample evidence exists to suggest that the immune system communicates with, and influences the nervous system both centrally and peripherally. Pro-inflammatory cytokines have been shown to influence the nervous system directly by altering ion channel kinetics, as well as indirectly by altering enzyme function thereby resulting in changes in critical neuroactive compounds. Proinflammatory mediators have been shown to up-regulate the enzyme indoleamine 2,3 dioxygenase (IDO) resulting in the accelerated degredation of serotonin precursor tryptophan (TRP) and increased production of TRP metabolites such as kynurenine (KYN). They have also been shown to upregulate the enzyme cyclooxygenase (COX) resulting in the increased production of pain inducing eicosanoids such as prostaglandin E2 (PGE2). Immune dysfunction in the form of chronic inflammation may therefore contribute to the severity of behavioural disorders such as depression and cognitive impairment, as well as neural disorders such neuropathic pain and somatic nerve deficits. SCI is typically associated with not only a state of chronic inflammation but also a drastically higher prevalence of each of the aforementioned neural and behavioural disorders. This makes SCI an ideal population to study the interaction between the immune and nervous systems, and assess the potential efficacy of novel treatment strategies which target the immune system for the management of such disorders. A 3-month anti-inflammatory diet was utilized as a treatment intervention for the purpose of reducing chronically elevated levels of pro-inflammatory mediators. This intervention allowed for the assessment of each of the outcome variables of interest at baseline (under an elevated inflammatory status) as well as at 1-month and 3-months during the intervention (under a reduced inflammatory state). Changes in inflammation were assessed by the quantification of serum pro (IL-1β, IL-2, IL-6, IFN-У, TNF-α, CRP) and anti-inflammatory (IL-4, IL-10, IL-1RA) cytokines. Cytokine-induced alterations in enzyme function and corresponding changes in neuroactive compounds were assessed by tryptophan (TRP), the competing amino acids phenylalanine (PHE), tyrosine (TYR), leucine (Leu), isoleucine (Ile), and valine (Val), the tryptophan metabolite kynurenine (KYN), and the pain-inducing eicosanoids prostaglandin E2 (PGE2) and leukotriene B4 (LTB4). In addition to such molecular indices, actual changes in each of the outcome variables of interest were assessed. Levels of depression were assessed by questionnaire via the Center for Epidemiological Studies Depression Scale (CES-D). Cognitive function (in the form of verbal learning) and memory was assessed via the California Verbal Learning Test (CVLT). Neuropathic pain was assessed via the Neuropathic Pain Questionnaire (NPQ). Somatic nerve function was assessed by EMG, including the assessment of nerve conduction velocity and signal amplitude in both motor and sensory nerves. The intervention significantly reduced serum concentrations of pro-inflammatory mediators in the treatment group (n=12) by 28%, while no significant change was found in the control group (n=8). Among other changes in amino acids, the most notable was that the change in the KYN/TRP ratio (an indicator of IDO activity) and the TRP/LNAA ratio (an indicator of TRP availability for serotonin synthesis) was significantly different between groups. The treatment group showed a significant reduction in scores of depression, as well as a significant reduction in sensory neuropathic pain scores. No significant changes were observed in regards to somatic nerve conduction and most indices of cognitive function (with the exception of the ability to avoid incorrect responses on the CVLT). These results may suggest a substantial role for chronic inflammation in depression and neuropathic pain following SCI and provide a potential alternative treatment strategy for the management of such intractable disorders.
    • New Pyrazole-Based Ligands and Their Complexes for Application in Transfer Hydrogenation and Hydrosilylation

      Alshakova, Iryna; Department of Chemistry
      A series of bidentate and tridentate ligands bearing pyrazolyl moiety in combination with phosphine, oxazoline, amine, and sulfide were synthesized. These ligands were applied for the synthesis of ruthenium complexes, that would be efficient in catalyzing transfer hydrogenation reaction in alcohol. From a number of obtained complexes, a mixture of two isomeric ruthenium complexes was found to be the most efficient in reduction of acetophenone and N-benzylideneaniline, as model substrates, with 2-propanol. These ruthenium complexes were successfully applied in transfer hydrogenation of nitriles, heterocyclic compounds, olefins, and alkynes. Activated esters were reduced under similar catalytic conditions when ethanol was used as a hydrogen source. These isomeric ruthenium complexes were also applied in the synthesis of secondary amines via hydrogen borrowing methodology. A number of primary amines and anilines were combined with primary alcohols under the conditions, optimized for transfer hydrogenation of nitriles, resulting in corresponding secondary amines. Furthermore, ammonium formate was used as a nitrogen source for alcohol amination. Thus, secondary and tertiary amines were obtained from primary alcohols. Another project was focused on transfer hydrogenation of carbonyl compounds with lithium isopropoxide. Addition of various ligands and small molecules was found to improve the reaction efficiency for aromatic substrates. Further studies revealed that lithium cation forms stable adduct with aromatic alcohols, while different additives help to break this interaction, thus resulting in significant improvement of the conversion to alcohols. Another strategy that was applied to improve the reaction yields was the addition of a cheap source of lithium cations, such as LiCl. Finally, a new zinc complex was synthesized and applied in the catalytic hydrosilylation of carbonyl compounds. The optimization of reaction conditions reviled that the presence of substoichiometric amounts of methanol in the system significantly accelerates the process. The reaction can proceed at very low catalyst load (down to 0.1mol%) under relatively mild reaction conditions. The substrate scope analysis showed the tolerance to carbon-carbon double bond. Thus, this procedure is efficient for the synthesis of allylic alcohols from α,β-unsaturated aldehydes and ketones.
    • New Synthetic Approaches and Structural Models of the Oxygen-Evolving Complex in Photosystem II from the Use of Oximato-Based Ligands

      Alaimo, Alysha; Department of Chemistry
      The employment of the chelating/bridging ligands salicylhydroxime (shiH3), quinoline-2-aldoxime (qaoH) and 2,6-diacetylpyridine dioxime (dapdoH2) in heterometallic Mn‒Ca chemistry has afforded various compounds with diverse topologies, metal stoichiometries and Mn oxidation state descriptions. Chapter 1 provides a general introduction to the oxygen-evolving complex (OEC) of Photosystem II (PSII) including discussions of fundamental aspects such as composition, structural proposals, mechanism of O‒O bond formation and synthetic approaches. My research results are reported in Chapter 2, 3 and 4. In the first project (Chapter 2), one-pot reactions between Mn(ClO4)2∙6H2O, Ca(ClO4)2∙4H2O and the potentially tetradentate chelating/bridging ligand salicylhydroxime (shiH3), resulting from the in situ metal ion-assisted amide-iminol tautomerism of salicylhydroxamic acid (shaH2), in the presence of various fluorescence carboxylate groups (2-naphthoic acid = L1-H; 9-anthracenecarboxylic acid = L2-H; 1-pyrenecarboxylic acid = L3-H) and base NEt3 has led to a family of structurally similar {MnIII4Ca} clusters (1‒4¬) with distorted square pyramidal topologies. The combined results demonstrate the ability of shiH3 and fluorescence carboxylates to yield new heterometallic Mn‒Ca clusters with (i) the same Mn‒Ca ratio as the OEC of PSII, (ii) structural stability in solution, (iii) a pronounced redox and optical activity and (iv) predominant antiferromagnetic exchange interactions with S = 0 spin ground states. These complexes may be relevant to lower oxidation level species of the catalytic cycle of the OEC. The second project of this thesis, discussed in Chapter 3, involved one-pot reactions between the [Mn3O(O2CPh)6(py)x]+/0 triangular precursors and either CaBr2∙xH2O or CaCl2∙6H2O in the presence of shaH2. This afforded the heterometallic complexes [MnIII4Ca2(O2CPh)4(shi)4(H2O)3(Me2CO)] (5) and (pyH)[MnII2MnIII4Ca2Cl2(O2CPh)7(shi)4(py)4] (6), respectively, in good yields. Further reactions but using a more flexible synthetic scheme comprising the Mn(NO3)2∙4H2O/Ca(NO3)2∙4H2O and Mn(O2CPh)2∙2H2O/Ca(ClO4)2∙4H2O “metal blends” and shaH2 in the presence of external base NEt3, led to the new complexes (NHEt3)[MnIII4MnIV4Ca2(OEt)2(shi)10(EtOH)2] (7) and (NHEt3)4[MnIII8Ca2(CO3)4(shi)8] (8), respectively. Solid-state dc magnetic susceptibility studies of 5‒8 revealed the presence of predominant antiferromagnetic exchange interactions between the Mn centers, leading to S = 0 spin ground state values. From a bioinorganic chemistry perspective, these compounds may demonstrate some relevance to both the high-valent scheme (7) and lower oxidation level species (5, 6 and 8) of the catalytic cycle of the OEC. In the last chapter of this thesis (Chapter 4), the ligands quinoline-2-aldoxime (qaoH) and 2,6-diacetylpyridine dioxime (dapdoH2) were introduced for a first time in heterometallic Mn‒Ca chemistry. This afforded a mixed-valence {MnII/III22Ca2} (9) cluster containing several {Mn4CaOx} subunits and a butterfly-like {MnIV2Ca2} (10) complex, respectively. These compounds demonstrate structural and magnetic relevance to both the low- and high-valent states of the OEC. All research-based Chapters (Chapter 2‒4) are divided into subsections in order to facilitate the understanding of the research concepts by the familiar and non-familiar readers and contextualize the messages, goals and conclusions of each individual project. I felt it was appropriate to begin each Chapter with a short preface of the work that summarizes the most important aspects of the specific project, followed by the complete experimental work and discussion of the results, and end with conclusions and some future perspectives.
    • Nonlinear dynamics of granular assemblies

      Przedborski, Michelle; Department of Physics
      In this work we investigate granular chains, which are one-dimensional systems of discrete macroscopic particles interacting via the intrinsically nonlinear Hertz law. Such systems support the propagation of solitary waves (SWs), which are non-dispersive, mobile bundles of energy. A comprehensive analysis into the dynamical behaviour of these systems and the properties of SW propagation is presented, and several interesting new results are obtained. First, we find that the transition to the quasi-equilibrium (QEQ) phase in granular chains can be manipulated by altering the material properties of the system. We further use these results to develop a novel shock absorption device with a predictable and tunable frequency response, making it useful also for energy harvesting applications. Second, we show for the first time that granular chains with various nonlinearities of the contact potential can achieve thermal equilibrium at sufficiently long times, and thus QEQ is an intermediate phase of these systems. We characterize the equilibrium phase by deriving approximate distribution functions for grain velocity and kinetic energy and system kinetic energy in a microcanonical ensemble of interacting particles. As a by-product, we derive the equilibrium specific heat, and a size-dependent correction term, for such systems. We also show how these ideas extend to heterogeneous systems such as diatomic, tapered, and random-mass chains. Furthermore, we look closely at the transition to equilibrium by using statistical tests to show that the long-term dynamics is ergodic, and by examining the behaviour of various correlation functions close to the onset of the transition. Third, we solve a highly nonlinear, fourth-order wave equation that models the continuum theory of long-wavelength pulses in weakly compressed, homogeneous granular chains with a general power-law contact interaction, to characterize all travelling wave solutions admitted by the equation. This involves deriving conservation laws admitted by the wave equation, followed by a modified energy analysis. We find that the wave equation admits various types of travelling wave solutions, including SW solutions as well as nonlinear periodic wave solutions. Not only have the SW solutions not appeared before in the literature on granular chains, but they are also a new addition to the literature on SWs in general.
    • Nonsuicidal Self-Injury and Suicidal Risk: An Examination among Young Adults

      Chloe, Hamza; Department of Psychology
      Nonsuicidal self-injury (NSSI), which refers to the direct and deliberate destruction of bodily tissue in the absence of suicidal intent, is a serious and widespread mental health concern. Although NSSI has been differentiated from suicidal behavior on the basis of non-lethal intent, research has shown that these two behaviors commonly co-occur. Despite increased research on the link between NSSI and suicidal behavior, however, little attention has been given as to why these two behaviors are associated. My doctoral dissertation specifically addressed this gap in the literature by examining the link between NSSI and several measures of suicidal risk (e.g., suicidal ideation, suicidal attempts, pain tolerance) among a large sample of young adults. The primary goal of my doctoral research was to identify individuals who engaged in NSSI at risk for suicidal ideation and attempts, in an effort to elucidate the processes through which psychosocial risk, NSSI, and suicidal risk may be associated. Participants were drawn from a larger sample of 1153 undergraduate students (70.3% female) at a mid-sized Canadian University. In study one, I examined whether increases in psychosocial risk and suicidal ideation were associated with changes in NSSI engagement over a one year period. Analyses revealed that beginners, relapsed injurers, and persistent injurers were differentiated from recovered injurers and desisters by increases in psychsocial risk and suicidal ideation over time. In study two, I examined whether several NSSI characteristics (e.g., frequency, number of methods) were associated with suicidal risk using latent class analysis. Three subgroups of individuals were identified: 1) an infrequent NSSI/not high risk for suicidal behavior group, 2) a frequent NSSI/not high risk for suicidal behavior group, and 3) a frequent NSSI/high risk for suicidal behavior group. Follow-up analyses indicated that individuals in the frequent NSSI/high risk for suicidal behavior group met the clinical cutoff score for high suicidal risk and reported significantly greater levels of suicidal ideation, attempts, and risk for future suicidal behavior as compared to the other two classes. Class 3 was also differentiated by higher levels of psychosocial risk (e.g., depressive symptoms, social anxiety) relative to the other two classes, as well as a comparison group of non-injuring young adults. Finally, in study three, I examined whether NSSI was associated with pain tolerance in a lab-based task, as tolerance to pain has been shown to be a strong predictor of suicidal risk. Individuals who engaged in NSSI to regulate the need to self-punish, tolerated pain longer than individuals who engaged in NSSI but not to self-punish and a non-injuring comparison group. My findings offer new insight into the associations among psychosocial risk, NSSI, and suicidal risk, and can serve to inform intervention efforts aimed at individuals at high risk for suicidal behavior. More specifically, my findings provide clinicians with several NSSI-specific risk factors (e.g., frequent self-injury, self-injuring alone, self-injuring to self-punish) that may serve as important markers of suicidal risk among individuals engaging in NSSI.
    • Novel glutathione disulfide transferase function of CC-glutaredoxins involved in disease resistance and flower development

      Slavickova, Nina; Centre for Biotechnology (Brock University, 2014-08-05)
      Glutaredoxins are oxidoreductases capable of reducing protein disulfide bridges and glutathione mixed disulfides through the process of deglutathionylation and glutathionylation. Lately, redox-mediated modifications of functional cysteine residues of TGA1 and TGA8 transcription factors have been postulated. Namely, GRX480 and ROXY1 glutaredoxins have been previously shown to interact with TGA proteins and have been suggested to regulate redox state of these proteins. TGA1, together with TGA2, is involved in systemic acquired resistance (SAR) establishment in the plant Arabidopsis thaliana through PR1 (Pathogenesis related 1) gene activation. They both form an enhanceosome complex with the NPR1 protein (non-expressor of pathogenesis related gene 1) which leads to PR1 transcription. Although TGA1 is capable of activating PR1 transcription, the ability of the TGA1 NPR1 enhanceosome complex to assembly is based on the redox status of TGA1. We identified GRX480 as a glutathionylating enzyme that catalyzes the TGA1 glutathione disulfide transferase reaction with a Km of around 20μM GSSG (oxidized glutathione). Out of four cysteine residues found within TGA1, C172 and C266 were found to be glutathionylated by this enzyme. We also confirmed TGA1 glutathionylation in vivo and showed that this modification takes place while TGA1 is associated with the PR1 promoter enzymatically via GRX480. Furthermore, we show that glutathionylation via GRX480 abolishes TGA1's interaction with NPR1 and consequently prevents the TGA1-NPR1 transcription activation of PR1. When glutathionylated, TGA1 is recruited to the PR1 promoter and acts as a repressor. Therefore, glutathionylation is a mechanism that prevents TGA1 NPR1 interaction, allowing TGA1 to function as a repressor of PR1 transcription. Surprisingly, GRX480 was not able to deglutathionylate proteins demonstrating the irreversible nature of the reaction. Moreover, we demonstrate that other members of CC-class glutaredoxins, namely ROXY1 and ROXY2, can also catalyze protein glutathionylation. The TGA8 protein was previously shown to interact with NPR1 analogs, BOP1 and BOP2 proteins. However, unlike the case of TGA1 NPR1 interaction, here we demonstrate that TGA8-BOP1 interaction is not redox regulated and that TGA8 glutathionylation by ROXY1 and ROXY2 enzymes does not abolish this interaction in vitro. However, TGA8 glutathionylation results in TGA8 oligomer disassembly into smaller complexes and monomers. Our results suggest that CC-Grxs are unable to reduce mixed disulfides, instead they efficiently catalyze the opposite reaction which distinguishes them from traditional glutaredoxins. Therefore, they should not be classified as glutaredoxins but as protein glutathione disulfide transferases.
    • Novel Magnetic Materials Based on Macrocyclic Ligands: Towards High Relaxivity Contrast Agents and Mononuclear Single-Molecule Magnets

      Stares, Emma; Department of Chemistry
      The preparation and characterization of coordination complexes of Schiff-base and crown ether macrocycles is presented, for application as contrast agents for magnetic resonance imaging, Project 1; and single-molecule magnets (SMMs), Projects 2 and 3. In Project 1, a family of eight Mn(II) and Gd(III) complexes of N3X2 (X = NH, O) and N3O3 Schiff-base macrocycles were synthesized, characterized, and evaluated as potential contrast agents for MRI. In vitro and in vivo (rodent) studies indicate that the studied complexes display efficient contrast behaviour, negligible toxicity, and rapid excretion. In Project 2, DyIII complexes of Schiff-base macrocycles were prepared with a view to developing a new family of mononuclear Ln-SMMs with pseudo-D5h geometries. Each complex displayed slow relaxation of magnetization, with magnetically-derived energy barriers in the range Ueff = 4 – 24 K. In Project 3, coordination complexes of selected later lanthanides with various crown ether ligands were synthesized. Two families of complexes were structurally and magnetically analyzed: ‘axial’ or sandwich-type complexes based on 12-crown-4 and 15-crown-5; and ‘equatorial’ complexes based on 18-crown-6. Magnetic data are supported by ab initio calculations and luminescence measurements. Significantly, the first mononuclear Ln-SMM prepared from a crown ether ligand is described.
    • Novel plant DNA binding protein: Non-expresser of pathogenesis related 1 gene (NPR1) involved in disease resistance

      Pirnia, Saba; Centre for Biotechnology
      For the first time, through validation of the modified Chromatin Immunoprecipitation (ChIP) method (in vitro ChIP), the direct binding of NPR1 to the PR1 promoter was demonstrated. This is a novel advancement on plant systemic acquired resistance (SAR)-mediated disease responses. The NPR1 protein (nonexpressor of pathogenesis related gene 1) is a transcriptional co-activator and positive regulator of SAR, a long-lasting mobile defense signal found in plants. The pathogenesis-Related gene 1 (PR1) is particularly induced during defense response, and as such, is typically used as a marker for establishment of SAR in plants. Salicylic acid (SA) is a phytohormone required for SAR-mediated defense responses against pathogens. Recently, the role of NPR1 as a SA receptor was demonstrated; SA has been shown to directly bind to NPR1 through Cysteine 521 and 529 on the Cterminus region of NPR1 via the transition metal copper. The binding of SA to NPR1 results in disruption of the interaction between BTB/POZ and the C-terminus domains of NPR1. Upon SA-NPR1 binding, the C-terminus transactivation domain is released from the auto-inhibitory BTB/POZ domain, resulting in activation of the NPR1 transcription co-activator function, followed by PR1 transcription in Arabidopsis thaliana. Arabidopsis thaliana has an inducible defense system and is considered a model plant for studying disease resistance responses. In the current research, NPR1 was demonstrated to bind to the PR1 promoter at two distinct regions, in the presence and absence of SA. In the presence of SA, the binding site of NPR1 was determined to be localized at the -636 to -646 base pair sequence; however, in the absence of SA, NPR1 was found to bind around the -790 to -833 base pair sequence. In addition, two distinct DNA binding domains were identified within NPR1, localized on the C and N-terminus regions. In the absence of SA, the DNA binding domain within the N-terminus region, located between amino acids 110-190, was shown to facilitate the binding of NPR1 to the PR1 promoter through the amino acid cysteine 150 (Cys150) via transition metal. The DNA binding domain on the C-terminus region, located between amino acids 513-535, was demonstrated to allow the binding of NPR1 to the promoter of PR1 in the presence of SA. Two amino acids, cysteine 521 and 529 (Cys521/529), were shown to be essential for SA binding to NPR1 and subsequent NPR1 binding to the PR1 promoter. Furthermore, 4hydroxy benzoic acid (4-OH-BA), the inactive analogue of SA, has been demonstrated to be a potent inhibitor of NPR1-PR1 promoter interaction, both in vivo and in vitro, by competing with SA for NPR1 binding. Moreover, we demonstrated that other analogues of the NPR1 protein, NPR2, NPR3, and NPR4, are also recruited to the PR1 promoter. NPR4 showed a similar binding profile to NPR1, both in the presence and absence of SA. NPR2 and NPR3 were observed to only interact with the PR1 promoter in the absence of SA. Both NPR5 and NPR6 were shown to forgo binding to the PR1 promoter, further confirming their role in plant developmental processes other than defense. In addition, the binding of NPR1 to the PR1 promoter was demonstrated to be conserved among other plant species, including rice and maize. Both rice and maize NPR1 proteins were observed to bind to the PR1 promoter in the presence of SA and a metal co-factor, similar to Arabidopsis NPR1. Our results expand our understanding of how NPR1 interacts with the PR1 promoter to regulate gene expression during SAR establishment. This study also revealed that NPR1-mediated SAR defense signaling is conserved among other crop species, which can potentially facilitate the identification of novel plant-priming compounds through high-throughput chemical screening procedures alongside the application of the validated in vitro ChIP technique as a primary screening method.
    • Novel ways to measure future-oriented cognition: Using parent-report measures and open-ended responses to explore young children’s future thinking development

      Mazachowsky, Tessa; Department of Psychology
      Future-oriented cognition encompasses a set of key abilities that children must develop for successful functioning in daily life including, saving, prospective memory, episodic foresight, planning, and delay of gratification. These future thinking abilities are supported by memory systems (e.g., semantic, episodic), as well as constructive processes, self-projection, and executive functions. Research primarily measures young children’s future-oriented abilities through behavioural tasks, which have various limitations and may not engage future thinking. The current studies introduce new methods to overcome some of these limitations: developing a parent-report questionnaire and examining children’s open-ended responses. In Study 1 (N = 101; Mazachowsky & Mahy, 2020), 3-to 7-year-old’s future thinking was examined to establish the psychometric properties of a new parent-report measure, The Children’s Future Thinking Questionnaire (CFTQ). The CFTQ detected development of children’s future thinking and is a reliable and valid measure. Study 2 (N = 48; Mazachowsky et al., 2020) examined 3-to 5-year-old children’s episodic foresight using a novel, open-ended version of the Picture-book task. Results showed that children were able to generate items for future use and were more successful with age. Children’s explanations for their generated items were typically present-focused and included both episodic and semantic details. Expanding on Study 2, Study 3 (N = 158; Mazachowsky et al., revisions requested) explored 3-to 5-year-old’s explanations for their item choices on two episodic foresight tasks to determine the degree to which these tasks engaged children’s episodic and future-oriented processes. Children provided more future-oriented explanations on the Picture-book task compared to the Spoon task, but episodicity did not differ between tasks. Further, children’s Picture-book task explanations included more first-person personal pronouns compared to the Spoon task, but explanations did not differ in other pronoun use. Together, these studies show that use of a parent-report measure and examination of children’s open-ended responses offer unique insight into the development of young children’s future thinking and engagement in future-oriented processes.
    • Novices' learning from the Internet : an exploration of navigation behaviours, learner-related factors, and mental effort

      Desjarlais, Malinda; Department of Psychology (Brock University, 2011-03-08)
      The current study was an exploration of why some novices are more successful than their peers when learning from the Internet by examining the relations among time spent with relevant information and changes in invested mental effort during Internet navigations as well as achievement. Navigation behaviours and learner characteristics were investigated as predictors of time spent with relevant information and changes in mental effort. Undergraduates (N = 85, Mage = 20 years, 5 months) searched the Internet for information corresponding to a low knowledge topic for 20 min while their eye gaze and pupil size were recorded. Pupil diameter was used as an objective, continuous measure of mental effort. Participants also completed questionnaires or computer tasks pertaining to s e l f-regulated learning characteristics (general intrinsic goal orientation and effort regulation) and cognitive factors (working memory control, distractibility and cognitive style). All analyses controlled for general mental ability, reading comprehension, topic and Internet knowledge, and overall motivation. A greater proportion of time spent with relevant information predicted higher scores on an achievement test. Interestingly, time spent with relevant information partially mediated the positive relation between the frequency of increases in invested mental effort and achievement. Surprisingly, intrinsic goal orientation was negatively related to time spent with relevant information and effort regulation was negatively related to the frequency of increases in invested mental effort. These findings have implications for supports when novices guide their own learning, especially when using the Internet.
    • De novo sequencing, annotation, and characterization of the genome of Lavandula angustifolia (Lavender)

      Nattamai Malli Pooranachandhiran, Radesh; Centre for Biotechnology
      Lavender (Lavandula angustifolia) is a perennial plant native to the Mediterranean region, best known for its essential oil (EOs) that have numerous applications in the pharmaceutical, cosmetic and perfume industries. We performed sequencing of the L. angustifolia genome and report a detailed analysis of the assembled genome, focusing on genome size, ploidy, and repeat content. The lavender genome was estimated to be around 870 Mbp (1C=0.96 pg) using a quantitative PCR method. Genome size was further validated through analysis of raw genome sequences using Kmergenie, providing a conclusive end to the lavender genome size dispute. The repeat element composition of the genome was analyzed using de novo (RepeatModeler) and library-based methods (RepeatMasker) and was estimated to be around 45% of the full genome or ~57% of the non-gap genome sequences. Further characterization revealed Long Terminal Repeat (LTRs) retrotransposons as the major repeat type, which contribute to ~18% of the genome, followed by DNA transposons at ~8.5% of the genome. Interestingly, unlike most other plant genomes, the lavender genome has many more Copia than Gypsy elements, both showing a trend of recent increasing activity. Furthermore, these LTRs, especially Copia elements, have shown active participation in gene function including genes for essential oil production, with Copia elements contributing to ~30 % of the coding DNA sequence (CDS) regions, in addition to promoter, intron and untranslated (UTR) regions. The lavender genome also has an unusually high number of miniature inverted-repeat transposable elements (MITEs) compared to other model plant genomes, with the number being ~88,000, which is close to that (~90,000) of the much larger maize genome. Analysis also revealed the lavender genome with a high proportion at polyploidy level, which is strongly biased towards regions containing essential oil genes, with polyploidization events in the lavender genome occurred between 16 to 41 Mya. In conclusion, our results reveal the lavender genome to be highly duplicated and with past and ongoing active retrotransposition, making the genome optimized for EO production.
    • Ojibwe Elders' Experiences of Peace: To Teach Our Well-Being With the Earth

      Lafleur, Gail Sarah; Department of Graduate and Undergraduate Studies in Education (Brock University, 2013-11-15)
      This research focuses on exploring the Anishinaabek/Ojibwe worldview founded upon the spiritual relationship with Mother Earth as the Anishinaabek view of peace to teach our well-being with earth. This research explores the experiences of four 21st century traditional Anishinaabek elders through describing their ways of knowing and of being as it relates to the Anishinaabek worldview of respect and peace with nature. This respect for Mother Earth and respecting earth’s way−akii-bimaadizi is articulated and shared regarding elders’ experiences of teaching our well-being with earth−Akinomaage mino akii-ayaa and is based upon Anishinaabek spirituality. This research details the Anishinaabek worldview from the elders’ shared experiences of earth as teacher and elder. Ten themes emerged from the data. These themes included (a) going back to our original gifts and instructions/building your sacred bundle/sharing your sacred bundle, (b) wisdom−nbwaakaawin: connecting the dots/original instructions/medicine−mshkiki/environmental consciousness, (c) sacred teachings/learning from the elders, (d) relationships/honoring elders/eldership, (e) political experiences and awareness, (f) a way of being in Anishinaabek research, (g) survival, (h) peace is our worldview demonstrated, (i) be aware of colonialistic thinking, (j) Akinomaage: earth as context. The researcher also shares her reflections as a researcher and as an Anishinaabekwe: Ojibwe woman.
    • On the Ball Implementation of Canada Basketball’s Athlete Development Model

      Whitaker-Campbell, Tammy; Applied Health Sciences Program
      The purpose of this dissertation was to explore the perceived benefits and challenges of Canada Basketball’s athlete development model (ADM)/long-term athlete development (LTAD) by administrators, learning facilitators, and coaches at Canada Basketball to better understand the barriers to and enablers of this process. The methodological approach used for the study was an exploratory case study. Methods were established that were consistent with the iterative nature of case study. In total, 5 participants who identified as administrator/learning facilitator/coach, 6 participants who identified as /learning facilitator/coach, and 1 participant who identified as a coach participated in the study. In-depth semi-structured interviews were conducted with each participant that provided new insight into participants’ perceptions of and experiences with ADM/LTAD relative to their positions. Analysis revealed themes related to perceived (a) benefits while using ADM/LTAD; and (b) and challenges with using ADM/LTAD. These findings provide a preliminary assessment of one sport specific athlete development model and may inform research of other sport-specific athlete development programs. Several implications of the study findings are discussed and suggestions are posed for future research.
    • Opening a can of worms : perceptions and practices of teachers in Newfoundland and Labrador incorporating the role of a therapist

      Maich, Kimberly.; Department of Graduate and Undergraduate Studies in Education (Brock University, 2009-05-28)
      Educational trends of inclusion and collaboration have led to changing roles of teachers, including an emphasis on personal support. To provide for social, emotional, and behavioural needs, teachers may adopt a therapeutic role. Many models for such support are proposed, with most models including the importance of student-teacher relationships, a focus on social, emotional, and behavioural development, and direct instruction of related skills. This study includes 20 interview participants. In addition, 4 of the 20 interview participants also took part in a case study. It examines whether participants adopt a therapeutic role, their beliefs about student-teacher relationships, whether they provide interventions in personal issues, and instructed social, emotional, and behaviour skills. Findings show that teachers adopt an academic role as well as a therapeutic role, believe student-teacher relationships are important, are approached about personal issues, and instruct social, emotional, and behavioural skills. Talking and listening are commonly used to provide support, typically exclusive of formal curricular goals. The challenges in providing front-line support issues that may be shared within an established student-teacher relationship are considered. Support in turn for teachers who choose to provide support for personal issues in the classroom within a therapeutic role are suggested, including recommendations for support and referral related to specific social, emotional, or behavioural scenarios that may arise in the school community.