CRISPR/Cas9-Mediated Gene Editing of ARG1 in Cell Lines and Primary Cells

Arginase 1 deficiency, a urea cycle disorder resulting from an inability of the body to convert arginine into urea, results in hyperargininemia and sporadic episodes of hyperammonemia. Arginase 1 deficiency can lead to a range of developmental disorders and progressive spastic diplegia in children, and current therapeutic options are limited. Clustered regularly interspaced short palindromic repeat (CRISPR) /CRISPR associated protein (Cas) 9 gene editing systems serve as a novel means of treating genetic disorders such as Arginase 1 (ARG1) deficiency, and must be thoroughly examined to determine their curative capabilities. In these experiments numerous guide RNAs and CRISPR/Cas9 systems targeting the ARG1 gene were designed and observed by heteroduplex assay for their targeting capabilities and cleavage efficiencies in multiple cell lines. The CRISPR/Cas9 system utilized in these experiments, along with a panel of guide RNAs targeting various locations in the arginase 1 gene, successfully produced targeted cleavage in HEK293, MCF7, A549, K562, HeLa, and HepG2 cells; however, targeted cleavage in human dermal fibroblasts, blood outgrowth endothelial cells, and induced pluripotent stem cells was not observed. Additionally, a CRISPR/Cas system involving partially inactivated Cas9 was capable of producing targeted DNA cleavage in intron 1 of ARG1, while a Cas protein termed Cpf1 was incapable of producing targeted cleavage. These results indicate a complex set of variables determining the CRISPR/Cas9 systems’ capabilities in the cell lines and primary cells tested. By examining epigenetic factors and alternative CRISPR/Cas9 gene targeting systems, the CRISPR/Cas9 system can be more thoroughly considered in its ability to act as a means towards editing the genome of arginase 1-deficient individuals.

Queen's University Journal (1897-1898)

The Journal has been Queen's main student newspaper since it was founded in 1873. It appears twice a week on campus with a mix of news, sports, and entertainment stories, editorials, letters to the editor, and photographs. The paper is students' most important source of news and general information and has been a training ground for scores of Canadian journalists.

Queen's University Journal (1898-1899)

The Journal has been Queen's main student newspaper since it was founded in 1873. It appears twice a week on campus with a mix of news, sports, and entertainment stories, editorials, letters to the editor, and photographs. The paper is students' most important source of news and general information and has been a training ground for scores of Canadian journalists.

Queen's University Journal (1899-1900)

The Journal has been Queen's main student newspaper since it was founded in 1873. It appears twice a week on campus with a mix of news, sports, and entertainment stories, editorials, letters to the editor, and photographs. The paper is students' most important source of news and general information and has been a training ground for scores of Canadian journalists.

EDUCATING THE PUBLIC THROUGH NEWS MEDIA: CASE STUDIES OF NEWS LITERACY IN THE NEW MEDIA LANDSCAPE

Two decades of unprecedented changes in the media landscape have increased the complexity of informing the public through news media. With significant changes to the way the news industry does business and the way news consumers access this information, a new set of skills is being proposed as essential for today’s news consumer. News literacy is the use of critical thinking skills to assess the reliability and source of the information that people consume on a daily basis, as well as fostering self-awareness of personal news consumption habits and how it can create audience bias. The purpose of this study was to examine how adults experience the news in their everyday lives and to describe the nature of the news literacy skills people employ in their daily news consumption. This study purposefully selected four adults who have completed high school, and who regularly consume news information across a number of platforms, both traditional and digital. Two of the participants, one man and one woman, were over 50 years old. One other male participant was in his 30’s and the final participant, a young woman, was in her 20’s. They all utilized both traditional and digital media on a regular basis and all had differing skill levels when using social media for information. Their news experiences were documented by in-depth interviews and the completion of seven daily news logs. In their daily logs the participants differentiated news information from other information available on-line but the interviews revealed a contradiction between their intentions and their news consumption practices. All four participants had trouble distinguishing between news and opinion pieces in the news information realm. In addition all but one seemed unaware of their personal bias and any possible effect it was having on their news consumption. Further research should explore the benefits of an adult-centered news literacy curriculum on news consumers similar to the participants, and should examine the development of audience bias and its relationship to the daily exposure people have to the torrent of information that is available to them on a daily basis.

DEVELOPMENT OF AN ADJUSTABLE SUTURE TECHNIQUE FOR TRABECULECTOMY

This research presents a new design of an adjustable suture that could provide a better intraocular pressure (IOP) control in the post treatment of trabeculectomy surgery and limit associated complication with the current suturing techniques. A better control in tension suture brings a great deal of advantages to this surgical technique compared with the traditional adjustable suture. A length adjustment can be added in advance to a 10-0 nylon suture which enables suture tension to be released during the postoperative period of trabeculectomy surgery. This adjustment has a D-ring geometry made of 10-0 nylon suture adhered to a 10-0 nylon surgical suture which is used to close the scalar flap. The D ring was adhered with about 180 microdroplet of Loctite 4311that was found to form a strong joint to connect the D ring to the main 10-0 nylon suture and strong enough to carry the added tension instead after cutting the central suture between the two joints of the D ring. The geometry of adjustment is the key factor of maintaining the IOP at the normal range and keeping the scleral flap tight enough and secure so that aqueous humor continues to percolate under the subconjunctiva. It has been found that a 365, and 450 µm length extensions can release suture tension postoperatively and relieve the intraocular pressure within the eye by 33, and 66% respectively. The fabrication process of the new adjustable suture was divided into two steps: fabrication of micro jig and forming microdroplets. A micro jig was fabricated in order to form and bond a precise length extension to the new design of the adjustable suture. In addition, a new liquid separation technique has been followed in this study in order to generate micro adhesive droplets as small as 50µm for bonding the new adjustable suture structure.

Queen's University Journal (1900-1901)

The Journal has been Queen's main student newspaper since it was founded in 1873. It appears twice a week on campus with a mix of news, sports, and entertainment stories, editorials, letters to the editor, and photographs. The paper is students' most important source of news and general information and has been a training ground for scores of Canadian journalists.

SINGLE STAGE LED DRIVER TECHNOLOGY WITH LOW FREQUENCY RIPPLE CANCELLATION

Because of high efficacy, long lifespan, and environment-friendly operation, LED lighting devices become more and more popular in every part of our life, such as ornament/interior lighting, outdoor lightings and flood lighting. The LED driver is the most critical part of the LED lighting fixture. It heavily affects the purchasing cost, operation cost as well as the light quality. Design a high efficiency, low component cost and flicker-free LED driver is the goal. The conventional single-stage LED driver can achieve low cost and high efficiency. However, it inevitably produces significant twice-line-frequency lighting flicker, which adversely affects our health. The conventional two-stage LED driver can achieve flicker-free LED driving at the expenses of significantly adding component cost, design complexity and low the efficiency. The basic ripple cancellation LED driving method has been proposed in chapter three. It achieves a high efficiency and a low component cost as the single-stage LED driver while also obtaining flicker-free LED driving performance. The basic ripple cancellation LED driver is the foundation of the entire thesis. As the research evolving, another two ripple cancellation LED drivers has been developed to improve different aspects of the basic ripple cancellation LED driver design. The primary side controlled ripple cancellation LED driver has been proposed in chapter four to further reduce cost on the control circuit. It eliminates secondary side compensation circuit and an opto-coupler in design while at the same time maintaining flicker-free LED driving. A potential integrated primary side controller can be designed based on the proposed LED driving method. The energy channeling ripple cancellation LED driver has been proposed in chapter five to further reduce cost on the power stage circuit. In previous two ripple cancellation LED drivers, an additional DC-DC converter is needed to achieve ripple cancellation. A power transistor has been used in the energy channeling ripple cancellation LED driving design to successfully replace a separate DC-DC converter and therefore achieved lower cost. The detailed analysis supports the theory of the proposed ripple cancellation LED drivers. Simulation and experiment have also been included to verify the proposed ripple cancellation LED drivers.

CERAMICS IN BRITAIN (1840-90): MEANINGS AND METAPHORS

The intention of this thesis, “Ceramics in Britain (1840–90): Meanings and Metaphors” is to present new approaches for interpreting ceramics in nineteenth-century Britain by situating, problematizing, and contextualizing pottery and porcelain in the popular debates of the day within the methodologies of material culture, design, cultural and art histories. I ask how did ceramics—portable, functional, and often decorative objects—contribute to shaping modes of experiences? Crockery, tableware and blue-white-porcelain, admittedly largely mediated in texts and paintings, are at the centre of this research to examine how they imposed symbolism and influenced the engagement of their subjects beyond their intended meanings and functions. This thesis tracks a common rhetoric shared by writers and artists across genres and understood by readers and viewers: crockery in the cupboard, on the mantel, the table or the floor were popular motifs exemplifying class, gender, character, etiquette, and taste. This thesis also seeks to map ceramics’ relations with other objects and people depicted. Their meanings and metaphors changed, depending on their exchange with other objects in the room and who uses them. The conventions of representing ceramics dictated a particular grammar that writers and artists used, critiqued, discarded or personalized. The examination of ceramics mediated in text and image especially in comparison with extant objects invites a deeper probing of both material culture and artistic practice, which helps to situate the agency of the ceramic objects themselves. Also this thesis, in attempt to explore new methodological approaches for ceramic studies, examines the social life of the mid-Victorian relief-moulded “Minster” Jug in the Gardiner Museum in Toronto. The product originating in Staffordshire in 1843 and exported to the colonies holds significance due to its multiple life histories. Viewing the “Minster” through the lenses of curator, collector, consumer, and critic its layered lives unfold to reveal the protocols of museum praxis as well as important aspects of mid-nineteenth-century British society related to design reform, gender, imperialism and consumption patterns. This thesis contends that the British experienced ceramics in sometimes unexpected ways, unrelated to their original purpose, such as tools of violence or containers of solace, and transformative fantasy.

Nonlinear soil-structure interaction for buried pressure pipes under differential ground motion

Pipelines extend thousands of kilometers across wide geographic areas as a network to provide essential services for modern life. It is inevitable that pipelines must pass through unfavorable ground conditions, which are susceptible to natural disasters. This thesis investigates the behaviour of buried pressure pipelines experiencing ground distortions induced by normal faulting. A recent large database of physical modelling observations on buried pipes of different stiffness relative to the surrounding soil subjected to normal faults provided a unique opportunity to calibrate numerical tools. Three-dimensional finite element models were developed to enable the complex soil-structure interaction phenomena to be further understood, especially on the subjects of gap formation beneath the pipe and the trench effect associated with the interaction between backfill and native soils. Benchmarked numerical tools were then used to perform parametric analysis regarding project geometry, backfill material, relative pipe-soil stiffness and pipe diameter. Seismic loading produces a soil displacement profile that can be expressed by isoil, the distance between the peak curvature and the point of contraflexure. A simplified design framework based on this length scale (i.e., the Kappa method) was developed, which features estimates of longitudinal bending moments of buried pipes using a characteristic length, ipipe, the distance from peak to zero curvature. Recent studies indicated that empirical soil springs that were calibrated against rigid pipes are not suitable for analyzing flexible pipes, since they lead to excessive conservatism (for design). A large-scale split-box normal fault simulator was therefore assembled to produce experimental data for flexible PVC pipe responses to a normal fault. Digital image correlation (DIC) was employed to analyze the soil displacement field, and both optical fibres and conventional strain gauges were used to measure pipe strains. A refinement to the Kappa method was introduced to enable the calculation of axial strains as a function of pipe elongation induced by flexure and an approximation of the longitudinal ground deformations. A closed-form Winkler solution of flexural response was also derived to account for the distributed normal fault pattern. Finally, these two analytical solutions were evaluated against the pipe responses observed in the large-scale laboratory tests.

A Poetics of Annotation: Alexander Pope's Footnotes

Studies of Alexander Pope's poetry tend to examine only the footnotes to his Dunciads, if they examine his footnotes at all. This dissertation will address this deficit in our understanding of Pope's poetics through an examination of Pope's use of footnotes in support of his verse throughout his career. With Gerard Genette's taxonomy of footnotes as variously paratext and text and Hugh Kenner's idea of the technological space of the printed page as frameworks within which Pope's footnotes operate, this dissertation will show that – over the course of his career – Pope developed a poetics of annotation that deployed footnotes rhetorically as appeals to ethos and pathos that both built up Pope's own authorial ethos for his audience in the literary market place of early eighteenth-century London and for posterity and used that authorial ethos in support of his literary and political friends.

The Flotation of Niobium Oxide Minerals from Carbonatite Ores

Global niobium production is presently dominated by three operations, Araxá and Catalão (Brazil), and Niobec (Canada). Although Brazil accounts for over 90% of the world’s niobium production, a number of high grade niobium deposits exist worldwide. The advancement of these deposits depends largely on the development of operable beneficiation flowsheets. Pyrochlore, as the primary niobium mineral, is typically upgraded by flotation with amine collectors at acidic pH following a complicated flowsheet with significant losses of niobium. This research compares the typical two stage flotation flowsheet to a direct flotation process (i.e. elimination of gangue pre-flotation) with the objective of circuit simplification. In addition, the use of a chelating reagent (benzohydroxamic acid, BHA) was studied as an alternative collector for fine grained, highly disseminated pyrochlore. For the amine based reagent system, results showed that while comparable at the laboratory scale, when scaled up to the pilot level the direct flotation process suffered from circuit instability because of high quantities of dissolved calcium in the process water due to stream recirculation and fine calcite dissolution, which ultimately depressed pyrochlore. This scale up issue was not observed in pilot plant operation of the two stage flotation process as a portion of the highly reactive carbonate minerals was removed prior to acid addition. A statistical model was developed for batch flotation using BHA on carbonatite ore (0.25% Nb2O5) that could not be effectively upgraded using the conventional amine reagent scheme. Results showed that it was possible to produce a concentrate containing 1.54% Nb2O5 with 93% Nb recovery in ~15% of the original mass. Fundamental studies undertaken included FT-IR and XPS, which showed the adsorption of both the protonized amine and the neutral amine onto the surface of the pyrochlore (possibly at niobium sites as indicated by detected shifts in the Nb3d binding energy). The results suggest that the preferential flotation of pyrochlore over quartz with amines at low pH levels can be attributed to a difference in critical hemimicelle concentration (CHC) values for the two minerals. BHA was found to be absorbed on pyrochlore surfaces by a similar mechanism to alkyl hydroxamic acid. It is hoped that this work will assist in improving operability of existing pyrochlore flotation circuits and help promote the development of niobium deposits globally. Future studies should focus on investigation into specific gangue mineral depressants and inadvertent activation phenomenon related to BHA flotation of gangue minerals.

British Romantic Criticism and the Fine Arts: A Study in Philosophical Theories of Literary Unity

This dissertation is an exploration of how a small but important group of Romantic critics, finding fault in the ideal of three unities developed by neoclassical Academicians and wrongly attributed to Aristotle, turned to the terminology and practices of the fine arts to emphasize their conception of organic unity in literature. The Romantic analogy to painting in particular enables a philosophical criticism of literature to present the aesthetic semblance of painting, the comprehension of a multitude of details in a harmonious whole that is a natural unity to its medium, as a paradigm of modern-romantic poetry and its aspirations to similar complexity, particularity, and imaginative colour. Further, in extension of the French Querelle des anciens et des modernes of the seventeenth century, the division of ancient and romantic art by Romantic critics like August Schlegel, Samuel Taylor Coleridge, and William Hazlitt not only establishes an ethnological and historical difference between the artistic productions of these two cultural periods but also allows, unlike the neoclassical unities, a non-anachronistic philosophical vocabulary of whole and parts or of the general and particular in the criticism of poetry, which involution provides a “rule” more consonant with the laws of the imagination rather than with the rhetorical and absolutist dicta that were thither available in the literary canon.

Stitching Settler Identities: Canadian Quilts and Their Makers, 1800-1880

Stitching Settler Identities: Canadian Quilts and their Makers, 1800-1880 explores the making, use, and circulation of handmade settler quilts as representation of nineteenth-century women’s social, cultural and economic histories in Canada. An important part of Canadian settlement history is the making and use of handmade quilts in the settler homestead. Handmade coverlets that provided both physical and emotional warmth in the home were a measure of a settler-woman’s careful management of resources and a display of her innovation and creativity. Few settler women recorded their daily experiences; however, most women could sew and quilts offered a method of expression that allowed them to reflect and portray their identities. Thus far, the few studies of quilts have been limited to exhibition catalogues or research that considers a quilt’s aesthetics or its historic significance. While several scholars have called for a reclassification of textile production and needle arts to advance the way in which settler women were viewed as social beings – creating, producing, communicating, and circulating cultural values, most studies on quilts have overlooked a coverlet’s materiality. This study aims to expand the research on quilts as material culture within the context of art history by also considering a quilt’s materiality and when possible, its maker's biography.

WAKING (TO) THINKING BEING: A STUDY OF EDUCATION, METAPHYSICS, MIND, AND LANGUAGE

This dissertation articulates the basic aims and achievements of education. It recognizes language as central to thinking, and philosophy and education as belonging profoundly to one another. The first step is to show that although philosophy can no longer claim to dictate the foundations of knowledge or of disciplines of inquiry, it still offers an exceptionally general level of self-understanding. Education is equally general and faces a similar crisis of self-identity, of coming to terms with reality. Language is the medium of thought and the repository of historical mind; so a child’s acquisition of language is her acquisition of rational freedom. This marks a metaphysical change: no longer merely an animal, she comes to exercise her powers of rationality, transcending her environment by seeking and expressing reasons for thinking and doing. She can think about herself in relation to the universe, hence philosophize and educate others in turn. The discussion then turns to the historical nature of language. The thinking already embedded in language always anticipates further questioning. Etymology serves as a model for philosophical understanding, and demonstrates how philosophy can continue to yield insights that are fundamental, but not foundational, to human life. The etymologies of some basic educational concepts disclose education as a leading out and into the midst of Being. The philosophical approach developed in previous chapters applies to the very idea of an educational aim. Discussion concerning the substantiality of educational ideals results in an impasse: one side recommends an open-­ended understanding of education’s aims; the other insists on a definitive account. However, educational ideals exhibit a conceptual duality: the fundamental achievements of education, such as rational freedom, are real; but how we should understand them remains an open question. The penultimate chapter investigates philosophical thinking as the fulfillment of rational freedom, whose creative insights can profoundly transform our everyday activities. That this transformative self-understanding is without end suggests the basic aims of education are unheimlich. The dissertation concludes with speculative reflection on the shape and nature of language, and with the suggestion that through education reality awakens to itself.