Recombinant cationic biopolymers for nucleic acid delivery

Delivery of nucleic acids in an efficient and safe manner remains an unmet need in the biological sciences. Whether this means using gene therapy to introduce functional genes to cells, or using RNA interference mechanisms to silence misregulated proteins, the main hindrance in these approaches to disease mitigation, is the lack of a suitable delivery platform. The first chapter is a literature review of the currently available technology for nucleic acid delivery including viral vectors as well as cationic lipids and polymers. Herein the advantages and disadvantages of both viral and non-viral delivery are discussed. In Chapter II, a biopolymer featuring a lysine-histidine (KH) condensing region fused to a targeting motif is discussed. The purpose of this study was to examine the effect of architecture on transfection efficiency of plasmid DNA through a series of in vitro biological and biochemical assays. Chapter III describes a recombinant biopolymer which features an arginine-histidine (RH) DNA condensing region, a targeting motif for HER2, and endosomal release fusogenic peptide, and a nuclear localization signal. Each domain was tested for functionality through in vitro assays. The multifunctional biopolymer demonstrated selective and efficient delivery of plasmid DNA and gene expression in SK-OV-3 cells. In Chapter IV the RH based biopolymers were characterized and evaluated in terms of their ability to deliver nucleic acids to either the cytoplasm or cell nucleus. Past delivery vectors have often emphasized extracellular barriers to entry and success meant internalization of the particle. Intracellular trafficking, however is also a key compenent in successful delivery. The objective of this study was to design a biopolymer that can be programmed via its amino acid sequence to deliver siRNA specifically to cytoplasm. By modifying the amino acid sequence, the same biopolymer can also be programmed to deliver pDNA to the cell nucleus. Intracellular trafficking was observed through fluorescent microscopy and assays were conducted to demonstrate either siRNA knockdown or delivery of a suicide gene/prodrug combination. In the research presented, we describe a non-viral biopolymer system that is able to overcome the extracellular and intracellular barriers to nucleic acid delivery

The control of blood flow in skeletal muscle arterioles

"December 2010.""A Thesis presented to the Faculty of the Graduate School at the University of Missouri--Columbia In Partial Fulfillment of the Requirements for the Degree Master of Science."Thesis supervisor: Dr. Steven Segal.Skeletal muscle arterioles dilate in response to application of acetylcholine (ACh), eliciting a conducted vasodilation (CVD) that travels along unbranched segments without decrement. CVD is known to entail cell-to-cell transmission of hyperpolarization along the endothelium via gap junction channels, a purely passive mechanism. In the present thesis I study CVD in bifurcating arteriolar networks, where the pathway for hyperpolarizing current expands compared to unbranched arterioles, to test for an active component to CVD. In a separate subset of arterioles, the effect of augmenting vasomotor tone on CVD was tested using elevated O2 or phenylephrine (PE) in the superfusion solution vs. control. Male C57BL/6 mice (n=13; 10-13 weeks old) were anesthetized with pentobarbital sodium (50mg/kg, intraperitoneal injection) and maintained at 37 [degrees]C. The cremaster was carefully exteriorized and spread onto a transparent Sylgard pedestal. The tissue was maintained at 34 [degrees]C with continuous superfusion of physiologic saline solution. Microiontophoresis of ACh evoked non-decremental CVD in both unbranched and bifurcating arterioles, supporting the role of an active component in CVD. Further, augmenting vasomotor tone with PE attenuated non-decremental CVD, whereas increasing vasomotor tone with elevated O2 did not alter CVD. In summary, arteriolar networks in the mouse cremaster muscle exhibit robust dilation to ACh, which conducts along arterioles and across branch points without decrement, suggesting the contribution of both active and passive mechanisms. In preparations exhibiting poor spontaneous vasomotor tone, elevated O2 can be used to improve resting tone without impacting CVD.Includes bibliographical references (pages 51-58)

Canine Haiku: Yellow Ball

Poetry by Julie Andreyev and Tom

An Evaluation of High School Chorus Materials

One of the most important problems of a chorus director is how to select suitable materials which will be effectively used with a particular group. This problem is worthy of careful consideration. It is the purpose of this thesis to determine the factors necessary in choosing good material for the high school chorus and to offer a list of the best available selections as an aid to the director in choosing the most appropriate music

The Place of Music in the Religious Education of Children

The purpose of this thesis is to point out the importance of music in the religious education of children and to encourage the church to become aware of its task and its possibilities with its music so that music becomes a more effective aid in religious training

Pengaruh Kontaminasi Hasil Korosi Logam dari Alat Ortodontik Cekat terhadap Alergi pada Rongga Mulut

The aim of this orthodontic literature survey is to know how far the Influence of metal corrosion from the orthodontic fixed appliances toward the allergic in the oral cavity. The.conclusion of this survey are :1). The metal of fixed appliances that are used in the mouth should be had of high resistance corrosion. Stainless steel is suitable for it, besides that it is cheaper than other metal alloy, 2). Orthodontic treatment that uses stainless steel alloy from nickel chromium is better than nickel titanium toward the possibility of allergic incidences . 3). There is a relationship between personal history of metal allergic reaction and uses metal object on the skin with hypersensitivity of using nickel in the mouth. 4). Severe gingivitis, ulcers in oral mucous of the patients of orthodontic treatment needs to have more attention the possibility of hypersensitivity reaction to nickel or chromium toward the component of fixed appliances. Key words : Corrosion, stainless steel, nickel, chromium, hypersensitivity, allergi

Classification of Intervertebral Disc Disease

Intervertebral disc disease (IVDD) has been recognized in dogs since the 1800s, when the first descriptions of extruded disc material within the vertebral canal were published. In the intervening time our understanding of intervertebral disc pathology in dogs and cats has increased dramatically, with many variations of IVDD described. Whilst the volume of literature and collective understanding of IVDD has expanded, there has also been scope for confusion as the definition of intervertebral disc disease, with its myriad different manifestations, becomes more complicated. A large volume of literature has aimed to combine the use of histopathology, diagnostic imaging and clinical findings to better understand the various ways in which IVDD can be classified. Much of this research has focused on the classification of mechanisms of intervertebral disc degeneration, centering around the differences between, and overlaps in, IVDD in chondrodystrophic and non-chondrodystrophic dog breeds. However, with the increasing availability of advanced imaging modalities allowing more accurate antemortem diagnosis, the concept of IVDD has expanded to include other clinical presentations that may not fit into traditional models of classification of IVDD. This review aims to provide an up to date overview of both historical and current systems of IVDD classification, highlighting the important findings and controversies underpinning them

Faulkner's theory of relativity : relative clauses in Absalom, Absalom!

The purpose of this study is twofold: first, to show that William Faulkner used relative clauses (RC's) to shape the style of Absalom, Absalom! to fit the meanings he wanted to communicate, and in doing so built in a level of meaning beyond the "story;" second, that linguistic analysis of literature can illustrate precisely how style and meaning are interrelated, and in doing so can provide data about linguistic performance, in this case about the uses of RC's. Faulkner's own comment that "the theme, the story, invents its own style," justifies this study of Absalom, Absalom! as a speech act, or communicative event involving the interaction of writer, reader, text, and unspoken "rules" governing language use and interpretation. Within this framework, it is assumed that Faulkner chose to write this novel in the particular style he did for some meaningful purpose that readers would infer from that form. Critics have noted that the structure of the book is that of a story re-told from multiple perspectives; that the themes include the impossibility of discovering definitive "truth," the difficulty of communication, and the effects of events over time that relate otherwise unrelated people; and that the complex syntax of the novel, with its accumulations of modifying clauses, attempts to simulate the flow of speech or thought