>Magister Scientiae - MSc / Human Immunodeficiency Virus (HIV), which is the cause of Acquired Immunodefiency Syndrome (AIDS) is a major global public health issue affecting over 37 million people worldwide and is responsible for claiming over 32 million lives since the discovery of the disease in 1981. Through effective diagnosis, treatment and prevention HIV is a manageable disease. Today, advanced antiretrovirals, known as HAART, serve as effective, first-line drug regimens, consisting of a variety of viral inhibitors, and have successfully helped viral suppression. However, issues arise with antiretrovirals due to patient non-adherence and the development of drug resistant mutations. Coupled with dormant HIV reservoirs, viral extinction is attenuated. It is therefore essential that effective alternative treatments are investigated. The exploration of nanomedicine for targeted drug delivery has shown an ability to prolong the drug circulation time, target drugs to specific sites in the body, and enhance drug effectiveness. A previous study demonstrated a novel therapeutic strategy that was based on a mutant version of the caspase-3 enzyme that can induce apoptosis in HIV infected cells. This therapeutic strategy has the potential to wipe out reservoirs of HIV infection. However, the therapeutic strategy lacked selectivity because the delivery mechanism was based on protein transduction technology which will result in the nonselective delivery of the drug. In this study, preliminary work towards the development of a targeted nanoparticle delivery system for this mutant caspase-3 enzyme is described. The study describes the synthesis of green fluorescent protein loaded alginate/chitosan nanoparticles that were functionalized with a DNA aptamer intended to target the nanoparticles to CD4 expressing cells, that are also targeted by HIV. The THP-1 cell line was used due to the ability of the cells to express CD4 receptors on the cell surface. The nanoparticles were synthesized through ionotropic gelation. The size, polydispersity, zeta potential and morphology were investigated by Dynamic Light Scattering and Scanning Electron Microscopy, respectively. The strongly negative zeta potential studies revealed stability of the nanoparticles in suspension and Scanning Electron Microscopy results showed an indicative collapse of the polymer network for the empty nanoparticles (i.e. nanoparticles not loaded with GFP), whereas solid, cuboid nanoparticles were shown for the GFP-loaded nanoparticles. Image-based fluorescence cytometry demonstrated that the GFP-loaded nanoparticles bind to the THP-1 cells that express the CD4 receptor. The results obtained are indicative of a potential drug delivery system for HIV treatment however, adjustments would need to be made to the current study to further develop this nanocarrier.

Dreiwertige Actinide (An(III)) und Lanthanide (Ln(III)) stellen im Falle ihrer Inkorporation eine ernste Gefahr für die Gesundheit des Menschen dar. An(III) sind künstlich erzeugte, stark radioaktive Elemente, die insbesondere bei der nuklearen Energiegewinnung in Kernkraftwerken entstehen. Durch Störfälle oder nicht fachgerechte Lagerung radioaktiven Abfalls können sie in die Umwelt und die Nahrungskette des Menschen gelangen. Ln(III) sind hingegen nicht radioaktive Elemente, die natürlicherweise vorkommen und für vielfältige Anwendungen in Technik und Medizin abgebaut werden. Folglich kann der Mensch sowohl mit An(III) als auch Ln(III) in Kontakt kommen bzw. sie inkorporieren. Es ist daher von enormer Wichtigkeit, das Verhalten dieser Elemente im menschlichen Körper aufzuklären. Während makroskopische Vorgänge wie Verteilung, Anreicherung und Ausscheidung bereits sehr gut untersucht sind, ist das Wissen hinsichtlich der chemischen Bindungsform (Speziation) von An(III) und Ln(III) in Körperflüssigkeiten noch sehr lückenhaft. In der vorliegenden Arbeit wurde daher erstmals die chemische Bindungsform von Cm(III) und Eu(III) in natürlichem menschlichem Urin (in vitro) spektroskopisch aufgeklärt und die gebildeten Komplexe identifiziert. Hierzu wurden auch grundlegende Untersuchungen zur Komplexierung von Cm(III) und Eu(III) in synthetischem Modellurin sowie mit den urinrelevanten organischen Modellliganden Harnstoff, Alanin, Phenylalanin, Threonin und Citrat durchgeführt und die noch unbekannten Komplexbildungskonstanten bestimmt. Abschließend wurden alle experimentellen Ergebnisse mit Literaturdaten und Vorherberechnungen mittels thermodynamischer Modellierung verglichen. Auf Grund der hervorragenden Lumineszenzeigenschaften von Cm(III) und Eu(III) konnte insbesondere auch die Eignung der zeitaufgelösten laserinduzierten Fluoreszenzspektroskopie (TRLFS) als Methode zur Untersuchung dieser Metallionen in unbehandelten, komplexen biologischen Flüssigkeiten demonstriert werden. Die Ergebnisse dieser Arbeit liefern damit neue Erkenntnisse zu den biochemischen Reaktionen von An(III) und Ln(III) in Körperflüssigkeiten auf molekularer Ebene und tragen zu einem besseren Verständnis der bekannten, makroskopischen Effekte dieser Elemente bei. Darüber hinaus sind sie die Grundlage weiterführender in-vivo-Untersuchungen.

Numerous metal release data have been published by the Div. Surface and Corrosion Science and the AISI 316L stainless steel particles’ behavior in artificial lysosomal fluid (ALF). This study aims to evaluate the effect of chemical components in ALF on metal release from stainless steel particles with a bottom-up methodology. Two sizes of 316L stainless steel particles were used to assess the particle size influence on the metal release in detail. The results show that organic complexing agents e.g. lactate, tartrate and citrate are responsible for the high metal release rate in ALF. Correlations between the metal release rate and the number of carboxyl groups of the organic ligand were observed. Moreover, metal release data in this study indicates no synergistic effects in ALF solution, and continued research is on-going to study the synergistic effects further. No quantitative rules of iron, chromium and nickel release from the same stainless particles could be found which may indicate that these metals are released through different pathways e.g. diffusion or chemical dissolution. In different solutions with different chemical components, one or more metal release mechanisms dominate over others and make the metal release rate unpredictable.

Cyclodextrin mediation offers a unique mechanism with the potential for interesting control of reaction parameters. Cyclodextrin mediation of hydrophobic monomers may offer desirable kinetics over conventional free radical polymerizations, and it has been shown in this work that cyclodextrin mediation facilitates polymerization of hydrophobic monomers in aqueous solution and in ethylene glycol. It also may be a facile method for controlling relative reactivity of comonomer mixtures. In addition, complexation of cyclodextrin with guest molecules has been utilized in selective synthesis where the host cyclodextrin has been utilized to sterically hinder the attack of certain reactive sites contained within the host cavity. This aspect of inclusion complexation could also be utilized in free radical polymerizations of monomers with multiple reactive double bonds to preferentially reduce the reactivity of the hindered reactive sites. This thesis involves the use of methylated(1.8)-beta-cyclodextrin (MeCD) as a mediator for polymerizations in solvents that would not facilitate polymerization of the pure monomer in the absence of cyclodextrin. This study focuses on the carbohydrate mediation of a series of methacrylic monomers. t-Butyl methacrylate, n-butyl methacrylate, cyclohexyl methacrylate, and 2-ethylhexyl methacrylate were complexed with methylated(1.8)-beta-cyclodextrin and subsequently dissolved in either water or ethylene glycol. The complexes were studied by 1H and 13C NMR spectroscopy, thin layer chromatography, CPK modeling, and thermogravimetric analysis, and were found to have molar ratios of cyclodextrin to monomer as high as 1.0 to 0.72. These complexes were then free radically polymerized in either water or ethylene glycol and resulted in high molecular weight polymers that precipitated out of solution, allowing for facile polymer isolation through filtration. Isolated yields were found to be as high as 86 %. The majority of the cyclodextrin remained in solution after polymerization. It was also recovered and found to be recyclable. Heterogeneous polymerizations were also performed with 2-ethylhexyl methacrylate in which linear dextrin and methylated (1.8)-beta-cyclodextrin were used in emulsifier quantities. It was found that linear dextrin, at concentrations of 3.0 wt% produced a stable latex product with high molecular weight and an isolated yield of >90%. MeCD on the other hand failed to produce a stable emulsion at concentrations between 0.9-3.0 wt%, but remarkably MeCD at 3.0 wt% gave high molecular weight coagulated polymer with a yield of >90%. It is proposed that a heterogeneous mechanism inconsistent with the four major types discussed by Arshady is taking place. Unlike typical suspension or emulsion polymerizations, the cyclodextrin mediated polymerizations are completely homogeneous at the onset, making them more like a dispersion or precipitation polymerization. However, in dispersion and precipitation polymerizations the pure monomer is soluble in the reaction media. In the absence of cyclodextrin, the monomers utilized in this study possessed no appreciable solubility in the reaction media. Therefore, it is proposed that cyclodextrin acts as a phase transfer agent, effectively solublizing the hydrophobic monomer and allowing for the aqueous dispersion or precipitation type polymerization to occur, depending on the relative solubility of the components. Bulk polymerizations of t-butyl methacrylate, cyclohexyl methacrylate, and 2-ethylhexyl methacrylate and their subsequent use in the preparation of carbohydrate/poly(alkyl methacrylate) blends was also performed in this project. Bulk polymers were utilized as references for physical properties for the polymers produced through polymerization of the MeCD/monomer complexes in either aqueous solution or in ethylene glycol. 1H NMR analysis of the polymers from both the cyclodextrin mediation and bulk polymerizations indicated that the tacticity of the polymers produced in both cases were identical. The bulk polymers were also used in the preparation of carbohydrate/methacrylic blends with potential applications in the areas of selective barriers, biodegradable films. Inclusion of drug molecules or antioxidants into these cyclodextrin containing films also may have potential in drug delivery, or food packaging applications. In addition, the side chain liquid crystalline monomer, 6-(4-hexyloxy-biphenyl-4-yloxy)hexyl methacrylate was synthesized in high purity via a three-step procedure and confirmed by a combination of mass spectrometry, thin layer chromatography, and 1H and 13C NMR. This hydrophobic liquid crystalline monomer was subsequently complexed with 1.0-3.0 equivalents of methylated(1.8)-beta-cyclodextrin in an attempt to alter the water solubility of the monomer. Complexes of this side-chain liquid crystalline monomer have not been studied previously and it is proposed that complexation with cyclodextrin will lead not only to novel polymerizations routes for this monomer, but also to novel smectic phases for this thermotropic liquid crystalline polymer. / Master of Science

Convergent and divergent strategies for the synthesis of viologen dendrimers with 1,3,5-tri-methylene branching units are presented. The synthesis of 3,5-bis(hydroxymethyl) benzyl bromide was optimized. The analysis of the crystal structure of 1-[3,5-bis(hydroxymethyl)benzyl]-4-(pyridin-4-yl) pyridinium hexafluorophosphate together with PM3 calculations opens an avenue to judge the structure and conformation of benzylic viologen dendrimers. In order to study chemical trigger induced conformational changes, viologen dendrimers were spin-labeled via a divergent approach. 1-(2,4-dinitrophenyl)-4-(pyridin-4-yl)pyridinium hexafluorophosphate was used as the end group to yield an activated dendrimer of the respective generation. The corresponding dendrimers were spin-labeled by reacting the active functionality with 4-amino TEMPO. The products were characterized by ESR (spin-label efficiency) and conventional cyclic voltammetry. Dynamic ESR studies are planned. New trimethylene-dipyridinium dendrimers were synthesized via a divergent approach using 4-tert-butylbenzyl group as the peripheral group. These dendrimers are well soluble in DMF or DMSO as PF6 salts and they act as a host for anthraquinone-2,6-disulfonate (AQDS). They can be stoichiometrically titrated with AQDS as shown by 1H-NMR, DOSY and cyclic voltammetry. Upon loading them with AQDS, the dendrimers undergo first a contraction, they reach a minimum hydrodynamic radius for complete charge compensation and they re-open when overcharging takes place. The contraction is supported by MM+ calculations. Upon stepwise loading of G2 (42 positive charges) with AQDS (2 negative charges), the first 3 molar equivalents (6 neg. charges) occupy the innermost dendrimer shell (consisting of 6 pos. charges), the next 6 equivalents (12 neg. charges) occupy the middle shell (12 pos. charges) and the last 12 equivalents AQDS (24 neg. charges) occupy the outermost shell of the dendrimer (24 pos. charges), as supported by 1H-NMR titrations yielding the magic equivalent numbers of 3, 9=3+6, and 21=3+6+12. Such stepwise radial complexations again in DMSO were further demonstrated using other molecular guests (mono-, di- and trianionic) as well as with on purpose synthesized viologen dendrimers. α,ω-dibromoalkanes were bifunctionalized in two steps to yield alkyl phosphonates with pyridinium, trimethylenedipyridinium, bipyridinium or a sulfonate at their ω end. These compounds were used as surface modifiers to build biomimetic membranes on the pore walls of mesoporous TiO2. Host-guest interaction studies with on purpose synthesized viologen compounds have been performed in collaboration.

Copper oxide nanoparticles (NPs) are used in an expanding range of industries including a potential for agricultural applications as a fungicide. Accidental spills or misapplication of CuO NPs may lead to soil contamination. Plant roots exude a wide range of organic chemicals for bioprotection and to enhance bioavailability of nutrients. Many of these chemicals are metal chelators that may increase the solubility of CuO NPs, thus enhancing the impact of these NPs on plants. This work was directed towards understanding which plant exudates force increased solubility of CuO NPs and to determine if the level of NP in the growth matrix drives a feedback effect, regarding composition and quantity of exudates. Wheat seedlings (Triticum aestivum cv Deloris) were grown in a sand matrix for 10 days after 3 days of germination. The sand was amended with sublethal doses of CuO NPs from 0 to 300 mg Cu/kg dry sand. Sand was selected as the solid growth matrix as a proxy for soil in terms of plant root morphology, mechanical impedance and water stress, while providing a low background of dissolved organic carbon for the isolation of root exudates. After plant growth, the pore water was collected from the sand by vacuum filtration and analyzed. By coupling analytic techniques including Triple Quad Mass Spectroscopy and ion chromatography with geochemical modeling, we have identified citrate and the phytosiderophore, deoxymugineic acid (DMA) as chelators that drove the majority of dissolution of CuO NPs, especially DMA at higher CuO NP doses. Altered biogeochemistry within the rhizosphere was correlated with increased plant uptake of Cu and bio-response via exudate type, quantity and metal uptake. Exposure of wheat to CuO NPs lead to dose-dependent reduction in Fe, Ca, Mg, Mn and K in roots and shoots. This work is relevant to growth of commercially important crop wheat in the presence of CuO NPs as a fertilizer, fungicide or a pollutant.

The purpose of this dissertation is to investigate the thermochemical properties of nonelectrolyte solutes dissolved in ternary solvent mixtures, and to develop mathematical expressions for predicting and describing behavior in the solvent mixtures. Forty-five ternary solvent systems were studied containing an ether (Methyl tert-butyl ether, Dibutyl ether, or 1,4-Dioxane), an alcohol (1-Propanol, 2-Propanol, 1-Butanol, 2-Butanol, or 2-Methyl-1-propanol), and an alkane (Cyclohexane, Heptane, or 2,2,4-Trimethylpentane) cosolvents. The Combined NIBS (Nearly Ideal Binary Solvent)/Redlich-Kister equation was used to assess the experimental data. The average percent deviation between predicted and observed values was less than ± 2 per cent error, documenting that this model provides a fairly accurate description of the observed solubility behavior. In addition, Mobile Order theory, the Kretschmer-Wiebe model, and the Mecke-Kempter model were extended to ternary solvent mixtures containing an alcohol (or an alkoxyalcohol) and alkane cosolvents. Expressions derived from Mobile Order theory predicted the experimental mole fraction solubility of anthracene in ternary alcohol + alkane + alkane mixtures to within ± 5.8%, in ternary alcohol + alcohol + alkane mixtures to within ± 4.0%, and in ternary alcohol + alcohol + alcohol mixtures to within ± 3.6%. In comparison, expressions derived from the Kretschmer-Wiebe model and the Mecke-Kempter model predicted the anthracene solubility in ternary alcohol + alkane + alkane mixtures to within ± 8.2% and ± 8.8%, respectively. The Kretschmer-Wiebe model and the Mecke-Kempter model could not be extended easily to systems containing two or more alcohol cosolvents.

Several recent studies that have proposed MUSIC models for gibbsite surfaces have purported to achieve a very good fit with potentiometric titration data. However, in order to achieve such results, several significant parameters, such as the number of surface sites, site densities, and pKa values were sometimes re-introduced in the model as fitted parameters, and physically unrealistic modeling assumptions were sometimes used. In addition, recent evidence supports the idea that some of the gibbsite potentiometric titration data from these studies may be unreliable. In order to re-interpret the potentiometric titration data, we used several recently published methods. In order to detect possible problems with estimates of gibbsite basal and edge surface area, we synthesized two gibbsite samples with different aspect ratios and characterized their surface areas using BET, AFM, and computerized image analysis routines. We also estimated pKa values for acid/base reactions at gibbsite surfaces by applying a new bond-valence method to gibbsite (001)-type and (100)-type surface structures based on ab initio calculations. The resulting pKa estimates are not to be taken as precise values due to difficulties and assumptions associated with calculating reasonable ab initio surface structures. Instead, we believe they represent a more reasonable range than has been previously estimated. Using these estimates, we propose an improved MUSIC model for gibbsite, which seems to predict surface adsorption, not perfectly, but within a reasonable range for a number of titration data sets without re-introducing any of our estimated parameters as adjustable parameters. Discrepancies that exist between model predictions and various potentiometric titration data sets are likely due to error associated with potentiometric titrations and pKa predictions.

Embargoed until July 2016. / Active Pharmaceutical Ingredients (APIs) are frequently prepared for delivery to the lung for local topical treatment of diseases such as Chronic Obstructive Pulmonary Disease (COPD) and asthma, or for systemic delivery. One of the most commonly used devices for this purpose is the pressurised metered dose inhaler (pMDI) whereby drugs are formulated in a volatile propellant held under pressure. The compound is aerosolised to a respirably sized dose on actuation, subsequently breathed in by the user. The use of hydrofluoroalkanes (HFAs) in pMDIs since the Montreal Protocol initiated a move away from chlorofluorocarbon (CFC) based devices has resulted in better performing products, with increased lung deposition and a concomitant reduction in oropharyngeal deposition. The physical properties of HFA propellants are however poorly understood and their capacity for solubilising inhaled pharmaceutical products (IPPs) and excipients used historically in CFCs differ significantly. There is therefore a drive to establish methodologies to study these systems in-situ and post actuation to adequately direct formulation strategies for the production of stable and efficacious suspension and solution based products. Characterisation methods have been applied to pMDI dosage systems to gain insight into solubility in HFAs and to determine forms of solid deposits after actuation. A novel quantitative nuclear magnetic resonance method to investigate the physical chemistry of IPPs in these preparations has formed the centrepiece to these studies, accessing solubility data in-situ and at pressure for the first time in HFA propellants. Variable temperature NMR has provided thermodynamic data through van’t Hoff approaches. The methods have been developed and validated using budesonide to provide limits of determination as low as 1 μg/mL and extended to 11 IPPs chosen to represent currently prescribed inhaled corticosteroids (ICS), β2-adrenoagonists and antimuscarinic bronchodilators, and have highlighted solubility variations between the classes of compounds with lipophilic ICSs showing the highest, and hydrophilic β2- agonist / antimuscarinics showing the lowest solubilities from the compounds under study. To determine solid forms on deposition, a series of methods are also described using modified impaction methods in combination with analytical approaches including spectroscopy (μ-Raman), X-ray diffraction, SEM, chromatography and thermal analysis. Their application has ascertained (i) physical form / morphology data on commercial pMDI formulations of the ICS beclomethasone dipropionate (QVAR® / Sanasthmax®, Chiesi) and (ii) distribution assessment in-vitro of ICS / β2-agonist compounds from combination pMDIs confirming co-deposition (Seretide® / Symbicort®, GlaxoSmithKline / AstraZeneca). In combination, these methods provide a platform for development of new formulations based on HFA propellants. The methods have been applied to a number of ‘real’ systems incorporating derivatised cyclodextrins and the co-solvent ethanol, and provide a basis for a comprehensive study of solubilisation of the ICS budesonide in HFA134a using two approaches: mixed solvents and complexation. These new systems provide a novel approach to deliver to the lung, with reduced aerodynamic particle size distribution (APSD) potentially accessing areas suitable for delivery to peripheral areas of the lung (ICS) or to promote systemic delivery.

Canada is in the process of implementing a Deep Geologic Repository (DGR) to dispose of used nuclear waste. Adsorption behaviour of both Se(-II) and Tc(IV) onto granite, shale, limestone, illite, and MX-80 bentonite has been elucidated. Se(-II) adsorption onto granite and MX-80 bentonite displays a decrease in Rd with an increase in solution pH. Se(-II) adsorption onto granite decreases with an increase in solution ionic strength. Se(-II) adsorption onto MX-80 bentonite does not return evidence which supports an apparent effect due to the ionic strength. Tc(IV) adsorption onto shale, limestone, illite, and MX-80 bentonite remains constant as the solution pH increases. Ionic strength does not affect the magnitude of Tc(IV) adsorption across the adsorbents, however an increase in ionic strength accelerates Tc(IV) adsorption. Se(-II) surface complexation models are best simulated with the following surface complexes: ≡Feldspar_sSe-, ≡Biotite_sOH2HSe, ≡Albite_sSe-, ≡Montmorillonite_sSe-, and ≡Montmorillonite_sOH2HSe. Tc(IV) adsorption is best simulated with: ≡Biotite_sOTcO(OH), ≡Quartz_sOTcO(OH), (≡Feldspar_sOH)2TcO(OH)-, ≡Montmorillonite_sOTcO(OH), (≡Albite_sOH)2TcO(OH)-, ≡Illite_sOTcO(OH), and ≡Chlorite_sOTcO(OH). Se(-II) adsorption onto granite and MX-80 bentonite in CR-10 solution returns Rd values of (1.80 ± 0.10) m3∙kg-1 and (0.47 ± 0.38) m3∙kg-1, respectively. Tc(IV) adsorption onto granite and MX-80 bentonite in CR-10 solution returned Rd values of (1.47 ± 0.25) m3∙kg-1 and (2.19 ± 0.33) m3∙kg-1, respectively. Tc(IV) adsorption onto shale, limestone, illite, and MX-80 bentonite in SR-270-PW solution returned Rd values of (0.16 ± 0.10) m3∙kg-1, (0.44 ± 0.21) m3∙kg-1, (1.86 ± 0.44) m3∙kg-1, and (0.23 ± 0.10) m3∙kg-1, respectively. This thesis will further deepen the understanding of Se(-II) and Tc(IV) adsorption. / Thesis / Doctor of Philosophy (PhD) / Determining the adsorption of Se(-II) and Tc(IV) onto granite, shale, limestone, illite, and MX-80 bentonite is beneficial to choosing a location within Canada to locate a used nuclear fuel repository. This thesis aims to quantify the adsorption behaviour of Se(-II) and Tc(IV) in Ca-Na-Cl and Na-Ca-Cl solutions with respect to a varying solution ionic strength and pH. Quantification of the adsorption was accomplished with adsorption experiments used in conjunction with geochemical simulations. New simulated surfaces specific to granite, shale, and MX-80 bentonite have been developed to complete these simulations. A final achievement was quantifying the adsorption of Se(-II) and Tc(IV) in groundwater representative solutions specific to locations considered for the used nuclear fuel repository.