Thesis (M.Com. (Industrial Psychology))--North-West University, Potchefstroom Campus, 2008.

Kochia and biennial wormwood are two weeds sometimes found growing in sunflower fields that may be difficult to control. Weed management in sunflowers is usually conducted using a combination of herbicides and mechanical weed control methods. Some farmers are growing sunflowers in reduced tillage systems, which may rely solely on herbicides to manage weeds; however, the spectrum of broadleaf weeds that can be controlled with herbicides is limited. Field experiments were conducted across southern Manitoba to determine the effect of kochia and biennial wormwood density and relative time of weed seedling recruitment on sunflower growth and development, yield and seed quality and to determine action thresholds for each weed. Early emerging kochia (plants that emerged at about the same time as the sunflowers) reduced sunflower yield by as much as 82%, which was greater than early emerging biennial wormwood plants, which reduced yield by as much as 27%. At low weed densities, each kochia plant reduced sunflower yield by 0.52% and each biennial wormwood plant reduced sunflower yield by 0.17%. As the density of early emerging kochia plants increased, sunflower height, stem diameter, leaf counts and head diameter were reduced in some of the experiments. Increasing densities of early emerging biennial wormwood plants had minimal effect on sunflower growth and development. Early emerging kochia and biennial wormwood plants both had the potential to reduce sunflower seed size and seed weight, while late emerging kochia and biennial wormwood (plants that emerged after the 4-leaf stage of the sunflowers) did not affect sunflower seed quality. The action threshold (5% sunflower yield loss) for early emerging kochia was 10 plants per metre square and the action threshold for early emerging biennial wormwood was 36 plants per square metre in the combined site-year analysis. Kochia or biennial wormwood plants that recruited after the 4-leaf stage of the sunflower crop did not affect sunflower yield, or seed quality.

Thesis (M.Com. (Industrial Psychology))--North-West University, Potchefstroom Campus, 2008.

This thesis is concerned with the measurement of the positions of points and bodies moving in trajectories in three dimensions, and the use of a new technique of optical interference which allows such measurements to be made dynamically. A variety of existing techniques for both static and dynamic three-dimensional position measurement are discussed, and the design of the new interferometer is introduced. The geometry of points, curves and surfaces in three dimensions is examined, with emphasis on the intersection of the point loci represented by the coordinate output of measuring instruments. The coordinates output by the interferometer represent surface loci which are quadric surfaces. A method of calculating the position and orientation of a body using three quadric surface intersection curves is presented. Diffraction of monochromatic light at an aperture is considered and it is shown that an interferometer working by division of wavefront can be used to obtain continuous information about the movement of the source of radiation, with that source free to move in up to three dimensions. A lens may be used to produce a compact instrument based on these principles. The diffraction integral equations are modified to incorporate the effect of a lens in the diffraction field. It is shown that even complex lenses can be represented by a few parameters in the diffraction equations. From the evaluation of these diffraction integrals, it is shown how the movement of interference fringes provides a coordinate output and how this is related to the locus of the radiation source. A method of obtaining very high resolution measurements of interference fringe pattern movement is presented. The interferometer was built and tested and the above theory verified in practice in a series of optical bench tests. The implementation of a system which uses this interferometer to measure the dynamic performance of industrial robots is considered. The optimum positions for the instruments are derived, and the method of designing the interferometer to give the required resolution is presented.

The ancient South American birds called tinamous (Tinamidae) have the smallest hearts known among birds and their cardiomyocytes have previously been shown to express significantly lower levels of the mitogen-activated protein kinase ERK compared to the more modern chicken (Gallus gallus). ERK is a well-known mediator of growth signalling in the heart, especially in hypertrophy. The aim of this project was to assess the effect of ERK knockdown on proliferation in cultured chicken cardiomyocytes. By transfecting these cells with a lipoplexed siRNA, ERK mRNA levels were knocked down to approximately half (45%, SD: 27%) compared to cells transfected with a negative control siRNA. The knockdown was coupled with a decreased proliferative response to insulin-like growth factor 1 (IGF-1) and foetal bovine serum (FBS). In conclusion, the ERK pathway was confirmed to be instrumental also in proliferative signalling. The results also support the notion that ERK itself is the rate-limiting step of this MAPK cascade. The low native expression of ERK in tinamou cardiomyocytes is expected to impose a strict limit on proliferative growth in response to various stimuli in these hearts. The genetic changes leading to higher expression levels, and with it the potential for larger hearts, in modern birds would have led to greatly increased evolutionary fitness by way of an increased aerobic scope and the ability to sustain flight.

Vesicular stomatitis virus (VSV) is considered a prototype for studying non-segmented negative-stranded ribonucleic acid (RNA) viruses. Livestock are naturally infected by VSV, causing severe economic impact due to lack of any effective treatment. RNA interference (RNAi)-based therapeutics are promising alternatives to control viral infections. Lentiviral vector systems deliver artificial short hairpin RNA (shRNA) into the genome of cells to activate the RNAi pathway. In this study, an RNAi-based approach to generate cell lines with reduced susceptibility to VSV (Indiana) infection was tested. First, eight shRNAs targeting either the nucleocapsid (N), phosphoprotein (P), or the polymerase (L) viral genes were designed and introduced into cell systems. To test the potency of the shRNAs for silencing the target viral transcripts, semi-quantitative polymerase chain reaction (PCR) analysis of viral N, P, and L transcripts was performed. Then, supernatants from infected groups were evaluated by microtitration and immunoblot. Finally, the effect of VSV genomic variability in the target region of shRNAs was predicted by partial sequencing field and laboratory-adapted strains. Viral transcripts were significantly reduced in cells stably expressing shRNAs targeting the N viral gene (nucleotides 67-97 or 1312-1332; p<0.05) or P gene (nucleotides 1772-1792; p&lt;0.05). Reduction in viral transcripts was not observed by other VSV-shRNAs tested. Reduction of viral transcripts by the N-shRNA (sh-1312) was accompanied by a decrease in viral protein. Also, a reduction in the viral particles shed from cells expressing N-shRNAs (nucleotides 67-97, p&lt;0.05) was noted. The results also showed complementarity of target gene sequences for shRNAs in the sequence from the laboratory-adapted strain and single base substitutions in the corresponding regions from VSV field isolates. However, these mismatches did not occur within the seed region of the shRNAs. In conclusion, partial silencing of viral transcripts by a single shRNA does not block VSIV replication; however, partial impairment of VSIV replication was observed in N-shRNAs expressing cells. During infection, the naturally high level of N gene transcription may have modulated the sh-RNA effect. The combination of the most potent shRNAs identified here into a multiple shRNA vector may result in further reduction of viral replication. These data contribute to ongoing development of effective RNAi-based technologies to combat viral diseases.

The ability of small interfering RNA (siRNA) to silence specific target genes offers not only a tool to study gene function but also represents a novel approach for the treatment of various human diseases, including cancers. The clinical use of siRNA, however, has been severely hampered by the inefficient delivery of these molecules to target cell populations due to their instability, inefficient cell entry, and poor pharmacokinetic profile. Much effort has therefore been devoted to the development of efficient in vivo siRNA delivery systems, with liposomes being the most widely employed vector. The traditional methods of packaging siRNA into liposomes, however, are often quite complex and labour-intensive, with the resulting products also being unstable at room temperature which limits their wide spread application in the clinic. The main aim of this research was to develop a simple, yet efficient, formulation technique to prepare stable siRNA-loaded liposomes which could be utilized as an efficient therapy for cancer treatment. Throughout this study, cervical cancer was used as the model system to assess the efficiency of various delivery systems. It is an ideal disease for siRNA therapy due to the cancer’s reliance on the expression of a single messenger RNA sequence which encodes two essential viral oncogenes, E6 and E7. Previous research has shown that targeting E6 and E7 by siRNA in cervical cancer cells in vitro results in either cell senescence or apoptosis. This thesis investigates the feasibility of applying E6/7 siRNA both intravaginally and intravenously to model the treatment of early-stage and end-stage cervical cancer, respectively. The practicability of applying E6/7 siRNA intravaginally for the treatment of localised cervical cancer tumours was firstly evaluated by administrating liposome-complexed siRNAs directly into the vaginal cavity of transgenic E7 mice. As no knockdown of E7 in cervical epithelium was observed for mice which received repeated treatments of E6/7 siRNA, the vaginal delivery efficiency of liposomes was further examined using fluorescently-labelled oligonucleotides. Contrary to previous reports, no delivery of lipoplexes into cervicovaginal tissues was detected irrespective of the dosage, type of lipid vector used, or the mouse estrus state at the time of administration. This lack of delivery was likely due to the poor retention of lipoplexes in the vaginal cavity as well as the inefficient penetration of lipoplexes across the mucosal layer lining the cervicovaginal epithelium. Overall, these findings indicated the necessity of developing more suitable and clinically acceptable vaginal siRNA delivery systems to enable this treatment strategy to become a reality. Despite the challenges of using liposomes to deliver siRNA via vaginal administration, their successful use in delivering siRNA intravenously to tumours was demonstrated in a subcutaneous cervical cancer mouse model. These experiments were carried out using PEGylated siRNA-loaded liposomes which were formulated using a novel Hydration-of-Freeze-Dried-Matrix (HFDM) technique. Compared to the existing formulation strategies, this method of preparation is less labour-intensive and the end product is also freeze-dried, ensuring product stability. It was found that the liposomes prepared using the HFDM method were stable in the presence of serum and they also possessed high siRNA entrapment and gene-silencing efficiencies. Following intravenous administration to mice, these particles were also found to accumulate in subcutaneous tumours to a similar degree compared to formulations prepared using a previously established technique. Importantly, these HFDM-formulated preparations showed superior stability over ones prepared using the traditional formulation method, with the particles still retaining 100% of their gene-silencing ability after storage for one month at room temperature. Using HFDM-formulated liposomes loaded with siRNA against Green Fluorescence Protein (GFP), a 50% knockdown of the GFP expression was achieved in tumours following intravenous administration. Additionally, the use of E6/7-targeted siRNA also resulted in a 50% reduction in tumour size when the siRNAs were delivered using HFDM-formulated liposomes. Importantly, this level of tumour growth suppression was comparable to that achieved from cisplatin, a clinically used chemotherapeutic for cervical cancer, at the clinically used dose. Overall, this research demonstrated that while there are still some challenges to overcome for siRNA to be used vaginally for cervical cancer treatment, HFDM-formulated PEGylated liposomes showed promise in bringing E6/7 siRNA forward as a treatment option for end-stage cervical cancer. In addition, the simplicity of preparation procedure along with superior product stability obtained from the HFDM method developed in this thesis will likely facilitate the translation of siRNA technology from laboratory to clinics for a range of other medical applications.

Thesis (Ph. D.)--University of Hong Kong, 2008. / Also available in print.

Thesis (M.S.)--Ohio University, August, 2001. / Title from PDF t.p.

Thesis (M.S.)--California Polytechnic State University, 2009. / Title from PDF title page; viewed on March 12, 2009. "February 2009." "In partial fulfillment of the requirements for the degree [of] Master of Science in Kinesiology." "Presented to the faculty of California Polytechnic State University, San Luis Obispo." Major professor: Kellie Green Hall, Ph.D. Includes bibliographical references (p. 43-45). Also available on microfiche.