Thesis (PhD)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Synthetic polymers are complex compounds that have multiple distributions with regard to molar mass, chemical composition, functionality and molecular architecture. Therefore, the molecular complexity of these compounds can only be analysed using a combination of analytical techniques. Well-defined complex polymers can be prepared by different types of living radical polymerisation, including reversible addition–fragmentation chain transfer polymerisation (RAFT) and reverse iodine transfer polymerisation (RITP). Using these techniques, several different homopolymers and copolymers have been prepared. However, there is still space for some more extended research. Many different types of multifunctional RAFT agents have been reported in literature. A tetrafunctional RAFT agent was prepared in our laboratory and used for the first time in the polymerisation of styrene. The polymerisation reaction was followed using in situ 1H nuclear magnetic resonance (NMR) and the molar masses of the resultant polymers were determined using size exclusion chromatography (SEC). The molar masses of the star-shaped polystyrenes (PS) were found to be less than the theoretical molar masses. This was due to the fact that SEC was calibrated with linear PS standards, while the samples under investigation are branched. Linear and branched polymers have different hydrodynamic volumes at similar molar masses. In order to prove that the star-shaped polymers were in fact four-armed, the samples were cleaved by aminolysis to yield the linear PS arms. The molar masses of the arms were in agreement with the theoretical arm molar masses based in the fourarmed structure. RITP is a relatively new living radical polymerisation technique. Various monomers have been prepared using RITP, including acrylates, methacrylates and styrene. The polymers formed using this technique have been characterised by techniques such as SEC, NMR and mass spectrometry (MS). However, very little advanced characterisation work has been done on polymers synthesised via RITP. Polystyrene-block-poly(n-butyl acrylate) (PS-b-PBA) block copolymers were prepared via RITP and the microstructure analysed by in situ NMR and other advanced analytical techniques. The chromatograms from gradient HPLC of the PS-b-PBA block copolymers showed a separation based on chemical composition. The preparation of deuterated polymers via RITP has not been reported in literature. Hydrogenous-polystyrene-block-deuterated-polystyrene (hPS-b-dPS) was synthesised via RITP and analysed using liquid chromatography at critical conditions. An isotopic separation was achieved when critical conditions were established for hydrogenous PS (h-PS). A separation of the block copolymer from the first block was also achieved under chromatographic conditions where the block copolymer eluted in SEC mode while the first block eluted in LAC mode. The separation according to the block structure was confirmed by two-dimensional liquid chromatography. / AFRIKAANSE OPSOMMING: Sintetiese polimere is komplekse verbindings wat meervoudige verspreidings ten opsigte van molêre massa, chemiese samestelling, funksionaliteit en molekulêre argitektuur. Daarom kan die molekulêre kompleksiteit van hierdie verbindings net ontleed word met behulp van 'n kombinasie van analitiese tegnieke. Goed-gedefinieerde komplekse polimere kan voorberei word deur verskillende soorte lewende radikaal polimerisasie, insluitend omkeerbare addisie-fragmentasie kettingoordrag polimerisasie (OAFO) en omgekeerde jodium oordrag polimerisasie (OJOP). Met behulp van hierdie tegnieke, was verskeie homopolimere en kopolimeer opgestel. Maar daar is nog plek vir nog uitgebreide navorsing. Baie verskillende tipes multifunksionele OAFO agente is aangemeld in die letterkunde. Ons het 'n nuwe vier-armige OAFO agent in ons laboratorium voorberei en dit was vir die eerste keer in die polimerisasie van stireen gebruik. Die polimerisasie reaksie is gevolg met behulp van in situ 1H kernmagnetieseresonans (KMR) en die molêre massas van die gevolglike polimere was bepaal deur grootteuitsluitings chromatografie (SEC). Die molêre massas van die ster-polistireen (PS) is bevind as minder as teoretiese molêre massas. Dit is omdat SEC instrumente gekalibreer word met lineêre PS standaarde, terwyl die monsters wat tans ondersoek word vertakte polimere is. Lineêre en vertakte polimere het verskillende hidrodinamiese volumes by soortgelyke molêre massas. Ten einde te bewys dat die ster polimere in werklikheid vier-armig is, is die monsters gesny deur ‘n aminolisasiereaksie om die lineêre PS arms te lewer. Die molêre massas van die arms was ooreenkomstig met die teoretiese arm molêre massas gebaseer op die vier-armige struktuur. OJOP is 'n relatiewe nuuts lewende radikaal polimerisasie tegniek. Verskeie monomere is opgestel deur OJOP, insluitend akrilate, metakrilate en stireen. Die polimere wat gevorm is deur middel van die tegniek is al gekenmerk deur tegnieke soos SEC, KMR en massaspektrometrie (MS). Tog is daar baie min gevorderde karakterisering werk gedoen oor polimere gesintetiseer deur middel van OJOP. Polistireen-blok-poli(n-butylacrylaat) (PS-b-PBA) blokkopolimere was voorberei deur middel van OJOP en die mikrostruktuur ontleed met behulp van gevorderde analitiese tegnieke. Die chromatogramme van gradiënt HPLC van die PS-b-PBA blokkopolimere het 'n skeiding ondergaan gebaseer op die chemiese samestelling. Die voorbereiding van gedeutereerde polimere deur middel van OJOP word nie in die letterkunde gevind nie. Gehidrogeneerde-polistireen-blok-gedeutereerde-polistireen (hPS-b-dPS) is gesintetiseer deur middel van OJOP en ontleed met behulp van vloeistofchromatografie onder kritiese kondisies. 'n Isotopiese skeiding was bereik wanneer kritiese kondisies gestig is vir gehidrogeneerde PS (h-PS).'n Skeiding van die blok kopolimeer van die eerste blok was ook bereik onder chromatografiese omstandighede waar die blok kopolimeer elueer in SEC terwyl die eerste blok elueer in LAC. Die skeiding volgens die blok struktuur was bevestig deur twee-dimensionele vloeistofchromatografie.

Thesis (MEng)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: FRC (Fibre-reinforced concrete) has become a common form of secondary and even primary reinforcing in some applications throughout the world. In South Africa, the structural applications are limited primarily to steel fibres while cheaper, lighter and more durable synthetic fibres have been side-lined due to low stiffnesses. The purpose of this research project is to investigate the behaviour of synthetic fibre-reinforced concrete (SynFRC) using fibres which are locally available in South Africa, and to propose a performance-based specification and test method for the use of these fibres. In order to achieve this, single fibre pull-out tests were performed on four locally available polypropylene fibres. It was found that the average bond stresses of the fibres are influenced primarily by the fibre cross sectional shape, longitudinal geometry and surface treatment, and secondarily by the aspect ratio. The W/C ratio had little effect on the single fibre performance of non-treated fibres, but appeared to have a slight effect on the single fibre performance of the surface treated fibre. From the experimental results, the highest fibre bond stress will be generated by using a fibre with an X-shaped cross section, longitudinal crimping and applying a surface treatment to this fibre. It also appears that the bond stress distribution for flat fibres is close to uniform, while the bond stress distribution for non-flat crimped fibres has a high mechanical interlock component at the surface end. Macro-mechanical performance tests were performed by means of the BS EN 14651 (2007) three point beam bending test and the ASTM C1550 (2012) Round Determinate Panel Test (RDPT). These tests were selected following a thorough literature review. The RDPT was found to be more consistent and able to identify trends which the three point beam bending test could not. In addition, the three point beam bending test’s most popular output, the Re,3 value tended to be misleading with varying W/C ratios, and it is recommended that the equivalent flexural tensile strength be used instead if the three point beam bending test is used. The macro-mechanical testing showed that increasing the fibre dosage did increase post-cracking performance. The flat fibres’ performance was significantly better than that of the non-flat fibres, and also increased at a faster rate with increasing fibre dosage. The post-cracking performance decreased with increasing W/C ratios and increasing aggregate sizes. The macro-mechanical performance was inversely proportionate to the single fibre performance. The macro-mechanical performance decreased with increasing fibre bond stress, and increased with increasing equivalent diameter, which equates to fewer fibres in a set volume of fibres. Finally, basic principles were developed from the data. These principles were used to predict the RDPT and three point beam bending test performance parameters based on fibre dosage, single fibre properties (bond stress and equivalent diameter), W/C ratio and aggregate size from the available data. The principles can be further refined with more experimental data. / AFRIKAANSE OPSOMMING: Vesel-gewapende beton word regoor die wêreld as ’n algemene vorm van sekondêre en selfs primêre versterking gebruik. In Suid-Afrika is die strukturele toepassings hoofsaaklik tot staal vesels beperk, terwyl goedkoper, ligter en meer duursame sintetiese vesels vermy word as gevolg van lae styfhede. Die doel van hierdie navorsingsprojek is om die gedrag van sintetiese-veselversterktebeton (SynFRC) te ondersoek deur gebruik te maak van vesels wat in Suid-Afrika beskikbaar is, en 'n prestasie-gebaseerdespesifikasie en toetsmetode vir die gebruik van sintetiese vesels voor te stel. Enkelveseluittrektoetse is op vier plaaslik beskikbare polipropileen vesels uitgevoer. Daar is gevind dat die gemiddelde verbandspanning van die vesel hoofsaaklik deur die vesel deursnee vorm, lengte meetkunde en oppervlak behandeling beïnvloed word, en tweedens deur die aspek verhouding beïnvloed. Die W/C-verhouding het min effek op die enkelveselprestasie van nie-behandelde vesels, maar het 'n effek op die enkeleveselprestasie van die oppervlak-behandelde vesel gehad. Die eksperimentele resultate wys dat die hoogste vesel verbandspanning deur 'n vesel met 'n X-vormige deursnit, lengte krimping en toepassing van 'n oppervlak behandeling gegenereer sal word. Dit blyk ook dat die verbandspanningverspreiding vir ’n plat vesel naby aan uniform is, terwyl die verbandspanningverspreiding vir ’n nie-plat gekrimpde vesel 'n hoë meganiese grendeling komponent op die oppervlak ente het. Makro-meganiese prestasietoetse is uitgevoer deur middel van die BS EN 14651 (2007) driepuntbalkbuigtoets en die ASTM C1550 (2012) RDPT. Hierdie toetse is ná ’n deeglike literatuuroorsig gekies. Die RDPT is meer konsekwent en is in staat om neigings te identifiseer wat die driepuntbalkbuigingtoets nie kan nie. Daarbenewens, met wisselende W/C verhoudings, is die driepuntbalkbuigtoets se gewildste resultaat, die Re,3-waarde geneig om misleidend te wees. Dit word aanbeveel dat die ekwivalentebuigtreksterkte in plaas van die Re,3-waarde as die drie punt balk buig toets resultaat gebruik word. Die makro-meganiesetoets het getoon dat die verhoging van die veseldosis ’n toename in na-krakingprestasie veroorsaak. Die plat vesels se prestasie was aansienlik beter as die van nie-plat vesels, en het met 'n toenemende veseldosis teen 'n vinniger koers verhoog. Die na-krakingprestasie het met toenemende W/C en die verhoging van die klip grootte afgeneem. Die makro-meganieseprestasie was omgekeerd eweredig aan die enkelveselprestasie. Die makro-meganieseprestasie het met toenemende vesel band stres verminder, en het met 'n toenemende gelykstaande deursnee (wat gelykstaande is aan minder vesel in 'n stel volume van vesel) vergroot. Ten slotte is basiese beginsels uit die data ontwikkel. Hierdie beginsels is gebruik om die RDPT en driepuntbalkbuigtoets prestasieparameters gebaseer op veseldosis, enkelveseleienskappe (verbandspanning en ekwivalentediameter), W/C-verhouding en klip grootte van die beskikbare data te voorspel. Die beginsels kan met meer eksperimentele data verder verfyn word.

In this dissertation, I design a processing approach, implement and test several solutions to combining spatial and spectral processing of multisource data. The measured spectral information is assumed to come from a multispectral or hyperspectral imaging system with low spatial resolution. Thematic content from a higher spatial resolution sensor is used to spatially localize different materials by their spectral signature. This approach results in both spectralunmixing and sharpening, a spatial-spectral fusion. The main real imagery example, fusion of polarimetric synthetic aperture radar (SAR) with hyperspectral imagery, poses a unique challenge due to the phenomenological differences between the sensors.Theoretical models for electro-optical image formation and scene reflectivity are shown to lead naturally to a set of pixel mixing equations. Several solutions for the spatial unmixing form of these equations are examined, based on the method of least squares. In particular, a method for introducing thematic content into the solution for spatial unmixing is defined using weighted least squares. Finally, and most significantly, a spatial-spectral fusion algorithm based on the theory of projection onto convex sets (POCS) is presented. Theoretical aspects of POCS are briefly discussed, showing how the use of constraints in the form of closed convex sets drives the solution. Then, constraints are derived that are intimately tied to the underlying theoretical models. Simulated imagery is used to characterize the different constraintcombinations that can be used in a POCS-based fusion algorithm.The fusion algorithms are applied to real imagery from two data sets, a Landsat ETM+ scene over Tucson, AZ and an AVIRIS/AirSAR scene over Tombstone, AZ. The results of the fusion are analyzed using scattergrams and correlation statistics. The POCS-based fusion algorithm is shown to produce a reasonable fusion of the AVIRIS/AirSAR data, with some sharpening of spatial-spectral features.

Magnetic resonance imaging (MRI) is an imaging technique that is used in hospitals worldwide. The images are acquired through the use of an MRI scanner and the clinical information is provided through the image contrast, which is based on the magnetic properties in biological tissue. By altering the scanner settings, images with different contrast properties can be obtained. Conventional MRI is a qualitative imaging technique and no absolute measurements are performed. At Center for Medical Imaging and Visualization (CMIV) researchers are developing a new MRI technique named synthetic MRI (SyMRI). SyMRI is based on quantitative measurements of data and absolute values of the magnetic properties of the biological tissue can be obtained. The purpose of this master thesis has been to take the development of SyMRI a step further by developing and implementing a visualization studio for SyMRI imaging of the human brain. The software, SyMRI Brain Studio, is intended to be used in clinical routine. Input from radiologists was used to evaluate the imaging technique and the software. Additionally, the requirements of the radiologists were converted into technical specifications for the imaging technique and SyMRI Brain Studio. Additionally, validation of the potential in terms of replacing conventional MRI with SyMRI Brain Studio was performed. The work resulted in visualization software that provides a solid formation for the future development of SyMRI Brain Studio into a clinical tool that can be used for validation and research purposes. A list of suggestions for the future developments is also presented. Future clinical evaluation, technical improvements and research are required in order to estimate the potential of SyMRI and to introduce the technique as a generally used clinical tool.

This thesis considers the use of Interferometric Synthetic Aperture Radar (InSAR) for surveying temporal fluctuations in the velocity of glaciers in the Arctic region. The aim of this thesis is to gain a broader understanding of the manner in which the flow of both land- and marine-terminating glaciers varies over time, and to asses the ability of InSAR to resolve flow changes over timescales which provide useful information about the physical processes that control them. InSAR makes use of the electromagnetic phase difference between successive SAR images to produce interference patterns (interferograms) which contain information on the topography and motion of the Earth's surface in the direction of the radar line-of-sight. We apply established InSAR techniques (Goldstein et al., 1993) to (i) the 925 km2 LangjÖkull Ice Cap (LIC) in Iceland, which terminates on land (ii) the 8 500 km2 Flade Isblink Icecap (FIIC) in Northeast Greenland which has both land- and marine-terminating glaciers and (iii) to a 7 000 km2 land-terminating sector of the Western Greenland Ice Sheet (GrIS). It is found that these three regions exhibit velocity variations over contrasting timescales. At the LIC, we use an existing ice surface elevation model and dual-look SAR data acquired by the European Remote Sensing (ERS) satellite to estimate ice velocity (Joughin et al., 1998) during late-February in 1994. A comparison with direct velocity measurements determined by global positioning system (GPS) sensors during the summer of 2001 shows agreement (r2 = 0.86), suggesting that the LIC exhibits moderate seasonal and inter-annual variations in ice flow. At the FIIC, we difference pairs of interferograms (Kwok and Fahnestock, 1996) formed using ERS SAR data acquired between 15th August 1995 and 3rd February 1996 to estimate ice velocity on four separate days. We observe that the flow of 5 of the 8 outlet glaciers varies in latesummer compared with winter, although flow speeds vary by up to 20 % over a 10 day period in August 1995. At the GrIS, we use InSAR (Joughin et al., 1996) and ERS SAR data to reveal a detailed pattern of seasonal velocity variations, with ice speeds in latesummer up to three times greater than wintertime rates. We show that the degree of seasonal speedup is spatially variable and correlated with modeled runoff, suggesting that seasonal velocity changes are controlled by the routing of water melted at the ice sheet surface. The overall conclusion of this work is that the technique of InSAR can provide useful information on fluctuations in ice speed across a range of timescales. Although some ice masses exhibit little or no temporal flow variability, others show marked inter-annual, seasonal and even daily variations in speed. We observe variations in seasonality in ice flow over distances of ~ 10 km and over time periods of ~10 days during late-summer. With the aid of ancillary meteorological data, we are able to establish that rates of flow in western Greenland are strongly moderated by the degree of surface melting, which varies seasonally and secularly. Although the sampling of our data is insufficiently frequent and spans too brief a period for us to derive a general relationship between climate and seasonality of flow, we show that production of meltwater at the ice surface and its delivery to the ice bed play an important role in the modulation of horizontal flow speeds. We suggest that a similarly detailed investigation of other ice masses is required to reduce the uncertainty in predictions of the future Arctic land-ice contribution to sea level in a warming world.

Membrane bioreactors (MBRs) show great promise for productivity improvement and energy conservation in conventional bioprocesses for wastewater reclamation. In order to attain high productivity in a bioprocess, it is crucial to retain the microorganisms in the bioreactors by preventing wash out. This enables recycling of the microorganisms, and is consequently saving energy. The main feature of MBRs is their permeable membranes, acting as a limitative interface between the medium and the microorganisms. Permeation of nutrients and metabolites through the membranes is thus dependent on the membrane characteristics, i.e. porosity, hydrophilicity,and polarity. The present thesis introduces membranes for MBRs to be used in a continuous feeding process, designed in the form of robust, durable, and semi-hydrophilic films that constitute an effective barrier for the microorganisms, while permitting passage of nutrients and metabolites. Polyamide 46 (polytetramethylene adipamide), a robust synthetic polymer, holds the desired capabilities, with the exception of porosity and hydrophilicity. In order to achieve adequate porosity and hydrophilicity, bulk functionalization of polyamide 46 with different reagents was performed. These procedures changed the configuration from dense planar to spherical, resulting in increased porosity. Hydroxyethylation of the changed membranes increased the surface tension from 11.2 to 44.6 mJ/m2. The enhanced hydrophilicity of PA 46 resulted in high productivity of biogas formation in a compact MBR, due to diminished biofouling. Copolymerization of hydrophilized polyamide 46 with hydroxymethyl 3,4-ethylenedioxythiophene revealed electroconductivity and hydrophilic properties, adequate for use in MBRs. To find either the maximal pH stability or the surface charge of the membranes having undergone carboxymethylation, polarity and the isoelectric point (pI) of the treated membranes were studied by means of a Zeta analyzer. The hydroxylated PA 46 was finally employed in a multilayer membrane bioreactor and compared with hydrophobic polyamide and PVDF membranes. The resulting biogas production showed that the hydroxylated PA 46 membrane was, after 18 days without regeneration, fully comparable with PVDF membranes.

In this thesis, the synthesis, reactivity and enantioselectivity of novel chiral Selectfluor Analogues were investigated. Chapter 1: Discussed is a general introduction to Selectfluor as an achiral electrophilic fluorinating reagent and its role as an oxidant in a wide variety of transformations and summarises the aims and objectives of the project. Chapter 2: The synthesis of chiral 2-monosubstituted, 2,3-disusbstituted and 2,5-disubstituted DABCO derivatives is described. We selected two strategies to prepare these chiral analogues of DABCO. Strategy A takes advantage of a large range of commercially available natural and unnatural amino acids. 2,3-Disubstituted DABCO motifs were prepared via a protocol of Sharpless in strategy B. Eleven chiral DABCO systems have been synthesized, six of which are novel compounds. Chapter 3 describes the synthesis of novel chiral non-racemic Selectfluor analogues using F₂. Mono-fluorinated chiral DABCOs were found to be unstable at low temperature, however, quaternized derivatives of these chiral reagents induced stability1. The fluorination proved challenging with chiral DABCO derivatives having sterically demanding substituents. Moreover, we have developed a practical and general process for the synthesis of novel chiral non-racemic Selectfluor using N-fluoropentachloropyridinium triflate 221. This methodology alleviates the need to handle F₂ gas. In Chapter 4: An investigation into how chiral Selectfluor analogues compare to N-F derived cinchona alkaloids in term of reactivity and enantioselectivity. The reactivity of the chiral Selectfluor analogues was challenged with both activated and less activated substrates. The novel chiral Selectfluor analogues showed superior reactivity than the cinchona alkaloids. Moderate to excellent yields of the desired products were obtained for all of the substrates studied.

The aim of this project was to utilize chiral cation-directed catalysis in the asymmetric synthesis of novel hererocycles. This goal was initially realized by the synthesis of azaindolines in high yields and enantioselectivities (Chapter 2). Extension of this methodology to substrates bearing two stereogenic centres was successful, although control over both diastereoselectivity and enantioselectivity in this process was modest. Finally the synthesis of heterocycles utilizing cation-directed rearrangement processes was examined, with proof of concept obtained for a novel asymmetric cyclization to form xanthenes.

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in Canada each year. Currently, most therapeutic approaches target rapidly dividing cancer cells by inhibition of normal cellular processes, however these therapies are not selective for cancer cells and unwanted side effects occur. Accordingly, novel cancer-targeted therapeutic strategies and drug targets are urgently needed to diminish the morbidity and mortality rates associated with CRC. Synthetic lethality is a new therapeutic approach that is designed to better target and kill cancer cells by exploiting a cancer-associated mutation (i.e. RAD54B-deficiency) thereby minimizing adverse side effects. We hypothesize that RAD54B-deficient CRC cells will be selectively killed via a synthetic lethal (SL) interaction with PARP1. We have identified and validated a novel drug target, PARP1, within CRC cells harboring RAD54B-deficiencies through a SL paradigm. This study represents the first steps necessary to identify and develop precision medicine based therapeutic strategies to combat CRC. / October 2016

By 2030, the global colorectal cancer burden is projected to approximately double. This highlights the immediate need to expand our understanding of the etiological origins of colorectal cancer, so that novel therapeutic strategies can be identified and validated. The putative tumor suppressor gene RNF20 encodes a histone H2B mono-ubiquitin ligase and has been found altered/mutated in colorectal and numerous other cancer types. Several studies suggest that RNF20, and by extension mono-ubiquitinated histone H2B (H2Bub1), play important roles in maintaining genome stability in human cells. Indeed, hypomorphic RNF20 expression and/or function have been shown to underlie several phenotypes consistent with genome instability, making aberrant RNF20 biology a potential driver in oncogenesis. Through an evolutionarily conserved trans-histone pathway, RNF20 and H2Bub1 have been shown to modulate downstream di-methylation events at lysines 4 (H3K4me2) and 79 (H3K79me2) of histone H3. Accordingly, understanding the biology associated with RNF20, H2Bub1, H3K4me2, and H3K79me2 is an essential preliminary step towards understanding the etiological origins of cancer-associated RNF20 alterations and identifying a novel therapeutic strategy to selectively kill RNF20-deficient cancers. In this thesis, I employ single-cell imaging, and multiple biochemical techniques to investigate the spatial and temporal patterning and characterize the biology of RNF20, H2Bub1, H3K4me2 and H3K79me2 throughout the cell cycle. In addition, I employ the CRISPR-Cas9 genome editing system to generate RNF20-deficient HCT116 cells. Finally, I employ synthetic lethal strategies to selectively kill RNF20-depleted cells. In conclusion, the research chapters contained within this thesis have characterized putative drivers in cancer (Chapter 3), generated a valuable research reagent for CRISPR-Cas9 ii genome editing experiments (Chapter 4), and identified a novel therapeutic strategy to selectively kill certain cancer cells (Chapter 5). This thesis has increased our understanding of the etiological origins of cancer and generated novel reagents and treatments strategies that after further validation and clinical studies, could be employed to reduce morbidity and mortality rates associated with cancer. / October 2016