D.Phil. (Chemistry) / Polycaprolactone (PCL), a linear, biodegradable polymer, and ethylene vinyl acetate (EVA), a branched copolymer, were used to prepare PCNs via the melt-blending method. Organoclay of the type Cloisite® 20A (C20A) and bentonite clay were used as fillers. The results show that the structure of a polymer matrix plays a significant role towards compatibilisation with the silicate layers of the clay. Scanning electron microscopy (SEM) and X-ray diffraction (XRD) analyses revealed an exfoliated-intercalated mixed morphology for the PCL matrix. However, for the EVA matrix, silicate layers agglomerated to form tactoids and resulted primarily in an intercalated morphology. Fourier transform infrared (FT-IR) spectroscopy was used to determine the nature of the interactions between the polymer and the filler. The thermal properties were investigated using thermogravimetric analysis (TGA) and indicated that, with an increase in clay loading, the thermal stability was reduced for both matrices, notwithstanding the type of polymer or clay used. Using EVA and C20A, this study revealed that more exfoliated nanocomposite structures can be obtained by using a modified solution-blending technique. This technique is a hybrid of the melt-blending and solution-blending methods. When compared to the melt-blending method, the modified solution method was found to be an efficient method for producing nanocomposite strips with uniform dispersion of the clay at organoclay loading of 8% and crystallinity by extrusion. However, the melt-blending method produced nanocomposites with high porosity, intercalation and thermal stability whereas the modified solution-blending technique resulted in more intercalated-exfoliated morphology, but less porosity and thermal stability. Despite the positives drawn from the modified solution method, the melt-blending method was used throughout for nanocomposites intended for application in water treatment. This was done because the solution used in the modified solution method could not be completely removed from the nanocomposite. Organic solvents can have a negative effect on the environment and human life.

Ph.D. (Chemistry) / The ultrafiltration membranes presented in this study were synthesized using the phase inversion method by casting on a nonwoven fabric. The polymer solutions were mixed with synthetic and bio additives in order to improve the resultant membrane performance. Synthetic additives (polyethylene glycol (PEG) and polyvinyl pyrrolidone (PVP)) were compared with a novel and cheap bio additive, lignin. Based on the knowledge that the additives must be soluble in water in order to increase the pore sizes of the membranes, sodium hydroxide was used to elute residual additives that remain in the membrane during coagulation. In order to trace the residual additives remaining, ATR-FTIR was used. Contact-angle analysis and water-absorption experiments were used to elucidate the hydrophilic properties of the prepared membranes. Membranes modified with lignin (Lig) were found to absorb more water (94% water uptake) when compared to the other membranes. In general, the contact angles were found to be low for membranes that were treated with NaOH. Membrane permeability followed the trend, Lig_PSf>PVP_PSf>PEG_PSf which is similar to the trend followed during water uptake. Pore size and pore distribution analysis showed that membranes modified with lignin and PVP had a narrow range (had pore sizes ranging from 10 to 24 nm) compared to that of PEG-PSf membrane (which ranged from 2.5 to 22 nm). A Robeson plot showed that Lig_PSf membranes had high separation factors regardless of the size of the solute being rejected. This study shows the possibility of using cheap and readily available additives to increase the performance of membranes......

No description available.

The enantioanalysis of compounds of clinical and pharmaceutical became increasingly important because the enantiomers of the same substance may be markers for different disease or are having a different pathway in the body. The utilization of enantioselective, potentiometric membrane electrodes made the assay of a single enantiomer easier and faster. Also the reliability of the analytical information is higher than that obtained using chromatographic techniques. The proposed electrodes are made by mixing graphite powder with paraffin oil to give carbon paste, which is modified by the addition of a chiral selector (e.g., cylodextrins, maltodextrins, macrocyclic antibiotics and fullerenes). This design is reliable. The high sensitivity, selectivity, enantioselectivity, accuracy and precision made them suitable to be used for the enantioanalysis of different compounds of clinical and pharmaceutical importance (e.g., L-histidine, L-cysteine and R-clenbuterol) in pharmaceutical tablets, and/or serum and urine samples. Copyright / Dissertation (MSc)--University of Pretoria, 2010. / Chemistry / unrestricted

No description available.

Aujourd'hui, la nanofiltration (NF) est perçue comme un procédé de séparation efficace et propre permettant d’extraire ou de concentrer des solutés ioniques ou neutres. Son développement à l’échelle industrielle n’est cependant pas encore optimal car les différents mécanismes de séparation sont encore mal compris. Cette thèse a porté d’une part sur l’étude des propriétés charge de surface des membranes de NF/OI et d’autre part sur les mécanismes de transport à travers des membranes de NF. Il a tout d’abord été montré, grâce à des mesures électrocinétiques réalisées sous atmosphère contrôlée, que certaines membranes commerciales subissent un post-traitement afin d’améliorer leurs performances de filtration. La détermination du potentiel zêta, grandeur caractéristique de la charge de surface, s’est révélée être une méthode adaptée pour mettre en évidence la présence d’une couche de post-traitement à la surface d’une membrane. La suite de l’étude a été consacrée à la compréhension des mécanismes de transport lors de la NF de mélanges ternaires contenant deux cations différents (Na+ et Ca2+) et un anion commun (Cl-). L’utilisation du modèle de transport SEDE a permis de montrer que les performances des membranes de NF résultent de la combinaison d’effets stériques, électriques et diélectriques. La dernière partie de ce travail a été consacrée à la NF d’un coproduit de l’industrie agro-alimentaire, le produit fermenté de soja (PFS). Il a été montré que la NF est potentiellement adaptée au dessalement du PFS (réduction de la teneur en cations monovalents) sans perte des sucres d’intérêt. / Today, nanofiltration (NF) is acknowledged as an efficient and environmentally-friendly separation process suited to extract or to concentrate neutral or ionic solutes. However, its development at industrial scale is not yet optimal because separation mechanisms are still poorly understood. This thesis focused on studying surface charge properties of NF/RO membranes and transport mechanisms through NF membranes as well. Firstly it was shown through electrokinetic measurements performed under controlled atmosphere that some commercial membranes undergo a post-treatment to improve their filtration performance. Determining the zeta potential, a physical quantity representative of the membrane surface charge, was shown to be a reliable method to highlight the presence of a post-treatment layer on the membrane surface. The second part of the study dealt with the understanding of transport mechanisms involved in NF of ternary mixtures containing two different cations (Na+ and Ca2+) and a common anion (Cl-). Using the SEDE transport model it was shown that the performance of NF membranes results from the combination of steric, electric and dielectric effects. The last part of this work was concerned with nanofiltration of a food industry co-product, the fermented soy product (PFS). It was shown that NF is potentially suitable for PFS desalination (reduction in the monovalent cation concentration) without loss of the sugars of interest.

The cystic fibrosis transmembrane conductance regulator (CFTR) is an ABC transporter family protein that acts as an ion channel. Mutations in CFTR cause the most common genetic disease in Caucasian populations, cystic fibrosis (CF). The high-resolution X-ray crystal structure of CFTR is now needed to aid the design of CFTR-targeted drugs for CF treatment and also to elucidate the molecular mechanisms behind its unique function as an ATP-ligand gated ion channel. However, until now, such structural studies have been severely limited by the lack of sufficient quantities of purified full-length CFTR protein. This thesis reports the novel over-expression and purification of milligram quantities of the chicken orthologue of CFTR protein from a Saccharomyces cerevisiae (yeast) expression system. A green fluorescent protein (GFP) tag fused to the CFTR C-terminus allowed rapid detection of the protein throughout the purification procedure. CFTR was expressed under an inducible promoter and appeared localised at, or near to, the plasma membrane, where it represented around 1 % of total protein after isolation in yeast microsomes. CFTR was solubilised from microsomes and purified using the detergents dodecylmaltoside (DDM) and lyso-phosphatidyl glycerol (LPG), by nickel affinity and size exclusion chromatography (SEC) to yield 1-2 mg of CFTR protein per 18 L fermentation culture. CFTR thermal stability was probed using fluorescent measurements to reveal a two-state cooperative unfolding transition around 40 °C for the DDM-purified protein, but no such transition was observed for the LPG-purified material. Light scattering and electron microscopy techniques revealed that, in LPG, CFTR was a homogenous population of monomeric particles around 60-Å in length that were soluble up to 8 mg/ml protein concentration. In DDM, CFTR was only soluble below 0.4 mg/ml protein concentration where is existed as a very heterogenous population of different sized amorphous particles, including dimeric particles around 180-Å in length. The DDM-purified CFTR protein could be crystallised as monomers in two-dimensional (2D) crystals with similar lattice parameters to 2D crystals of CFTR purified from mammalian cells. The ATPase activity of DDM-purified and reconstituted CFTR was similar to already published rates, at around 13 nmol Pi/min/mg integrated over a reaction time of 60 min, with an apparent affinity Km for ATP of 0.14 mM. Such a low ATPase rate compared to other ABC transporters may be due to the observed rapid run-down of activity with time and correlation with published CFTR channel gating kinetics. CFTR showed reduced ATPase activity after purification in LPG, suggesting a structural destabilisation in this detergent. The protocols presented here can now be used to provide sufficient quantities of purified CFTR protein for novel biochemical and biophysical studies. The tendency of CFTR to aggregate in a mild detergent remains a major obstacle towards 3D crystallisation trials and a high-resolution structure.

This dissertation the syntheses of Pt-based binary and ternary alloy electrocatalysts using the transition metals of Co and Ni are presented. These electrocatalysts were synthesised by an impregnation-reduction procedure at high temperature whereby Pt supported on carbon, (Pt/C (40 percent), was impregnated with the various metal and mixtures thereof and reduced at high temperatures in a H2 atmosphere. The procedure was also designed in such a way so as to prevent the oxidation of the support material (carbon black) during the alloy formation. The resultant nanoparticles (9-12 nm) of Pt3Co/C, Pt3Ni/C and Pt3Co0.5Ni0.5/C were also subjected to a post treatment procedure by acid washing (denoted AW) to produce electrocatalysts of Pt3Co/C-AW, Pt3Ni/C-AW and Pt3Co0.5Ni0.5/C-AW to study the effect of acid treatment on these electrocatalysts. The synthesised electrocatalysts were then characterised by a number of physical and electrochemical techniques and compared to that of commercial Pt/C (Pt/C-JM, HiSpec 4000) as well as Pt/C catalysts (Pt/C-900 and Pt/C-900-AW) treated under the same conditions used for the alloy synthesis. The electrocatalysts were then used to fabricate MEAs that were loaded into commercial single test cells and characterised by means of polarisation curves and Electrochemical Impedance Spectroscopy (EIS). The extensive physical characterisation included Powder X-Ray Diffraction (PXRD) analysis, Transmission Electron Microscopy (TEM), elemental analysis by Energy Dispersive Spectroscopy (EDS) and metal loading by Thermo-Gravimetric Analysis (TGA). These studies showed that Pt-based alloy electrocatalysts were successfully synthesised with particle sizes ranging from 9 - 12 nm, within their respective atomic ratios and whereby no significant loss of carbon support occurred. This indicated that significant sintering or electrocatalyst particles occurred when compared to that of the starting Pt/C catalyst (3 – 4 nm). From the combined results of the physical characterisation procedures, it was also shown that leaching as a result of acid washing was catalyst dependent with Ni containing catalysts showing a significant degree of leaching compared to that of Co containing catalysts. Electrochemical characterisation in terms of Electrochemical Active Surface Area (ECSA) by Cyclic Voltammetry (CV) and ORR activity by Rotating Disc Electrode (RDE) analysis revealed that a significant decrease in the ECSA resulted from the increase in particle size and this had a major influence on the ORR activity. Furthermore it was found that a significant improvement in the ORR activity was achieved by the synthesis of Pt-based alloys. It was also found that catalytic properties of the acid washed electrocatalysts were substantially different from that of non-acid washed electrocatalysts. The experimental data confirmed that it was possibly to achieve better catalytic performance as compared to that of Pt/C at a lower material cost when Pt is alloyed with base transition metals. The trend observed from the ORR activity studies by RDE was successfully repeated in the in-situ fuel cell testing in terms of mass activity of the electrocatalysts. Of the electrocatalysts studied under „real‟ fuel cell conditions Pt/C-JM had the best performance compared to the others, with the ternary Pt3Co0.5Ni0.5/C showing better catalytic performance compared to the Pt3Co/C electrocatalyst. This was found to be due to a higher charge transfer resistance observed in Pt3Co/C as compared to that of Pt3Co0.5Ni0.5/C which was similar than that of the commercial Pt/C-JM catalyst with both Pt3Co/C and Pt3Co0.5Ni0.5/C-AW having similar but higher ohmic resistances than that of Pt/C-JM as determined by electrochemical impedance spectroscopy. The results showed that a great potential exist to improve the catalytic performance of low temperature PEM fuel electrocatalysts at a reduced cost as compared to that of pure Pt provided a method of controlling the particle size was established.

Membrane proteins roughly constitute 30% of open reading frames in a genome and form 70% of current drug targets. They are classified as integral, peripheral membrane proteins and polypeptide toxins. α-helices and β -strands are the principal secondary structures observed in integral membrane proteins. This thesis presents the results of studies on analysis and correlation of sequence and structure of helices constituting integral helical membrane proteins. The aim of this work is to understand the helix stabilization, distortion as well as packing in terms of amino acid sequences and the correlated structures they adopt. To this end, analyses of datasets of X-ray crystal structures of integral helical membrane proteins and their comparison with a dataset of representative folds of globular proteins was carried out. Initial analysis was carried out using a non-redundant dataset of 75 membrane proteins to understand sequence and structural preferences for stabilization of helix termini. The subsequent analysis of helix distortions in membrane proteins was carried out using an updated dataset of 90 membrane proteins. Chapter 1 of the thesis reviews experimental as well as theoretical studies that have provided insights into understanding the structure of helical membrane proteins. Chapter 2 details the methods used during the course of the present investigations. These include the protocol used for creation of the non-redundant database of membrane and globular proteins. Various statistical methods used to test significance of the position-wise representation of amino acids in helical regions and the differences in globular and membrane protein datasets have been listed. Based on the tests of significance, a methodology to identify differences in propensity values that are statistically significant among two datasets has been devised. Programs used for secondary structure identification of membrane proteins namely Structure Identification (STRIDE) and Assignment of Secondary Structure in Proteins (ASSP) as well as those used for characterization of helical geometry (Helanal-Plus) have also been enlisted. In Chapter 3, datasets of 865 α-helices in 75 membrane proteins and 2680 α- helices from 626 representative folds in globular proteins defined by the STRIDE program have been analyzed to study the sequence determinants at fifteen positions within and around the α-helix. The amino acid propensities have been studied for positions that are important for the process of helix initiation, propagation, stabilization and termination. Each of the 15 positions has unique sequence characteristics reflecting their role and contribution towards the stability of the α-helix. A comparison of the sequence preferences in membrane and globular proteins revealed common residue preferences in both these datasets confirming the importance of these positions and the strict residue preferences therein. However, short/medium length α-helices that initiated/terminated within the membrane showed distinct amino acid preferences at the N-terminus (Ncap, N1, N2) as well as the C-terminus ( Ccap, Ct) when compared to α-helices belonging to membrane and globular proteins. The sequence preferences in membrane proteins were governed by the helix initiating and terminating property of the amino acids as well as the external environment of the helix. Results from our analysis also conformed well with experimentally tested amino acid preferences in a position-specific amino acid preference library of the rat neurotensin receptor (Schlinkmann et al (2012) Proc Natl Acad Sci USA 109(25):1890-5) as well as crystal structures of GPCR proteins. In the light of the environment dependent amino acid preferences found at α- helix termini, a survey was carried out to find various helix capping motifs adopted at both termini of α-helices in globular and membrane proteins to stabilize these helix termini. The results from these findings have been reported in Chapter 4. A sequence dependent structural preference is found for capping motifs at helix termini embedded inside and protruding outside the membrane. The N-terminus of α-helices was capped by hydrogen bonds involving free main chain amide groups of the first helical turn as donors and amino acid side chains as acceptors, as against the C-terminus which showed position-dependent characteristic backbone conformations to cap the helix. Overall helix termini inside the membrane did not show a very high number of capping motifs; instead these termini were stabilized by helix- helix interactions contributed by the neighboring helices of the helical bundle. In Chapter 5, we examine transmembrane helical (TMH) regions to identify as well as characterize the various types of helix perturbations in membrane proteins using ASSP and Helanal-Plus. A survey of literature shows that the term ‘helix kink’ has been used rather loosely when in fact helical regions show significant amounts of variation and transitions in helical parameters. Hence a systematic analysis of TMH regions was undertaken to quantify different types of helix perturbations, based on geometric parameters such as helical twist, rise per residue and local bending angle. Results from this analysis indicated that helices are not only kinked but undergo transitions to form interspersed stretches of 310 helices and π-bulges within the bilayer. These interspersed 310 and π-helices showed unique sequence preferences within and around their helical body, and also assisted in main- taining the helical structure within the bilayer. We found that Proline not only kinked the helical regions in a characteristic manner but also caused a tightening or unwinding in a helical region to form 310 and π-helix fragments respectively. The helix distortions also resulted in backbone hydrogen bonds to be missed which were stabilized by hydrogen bonds from neighboring residues mediated by their side chain atoms. Furthermore, a packing analysis showed that helical regions with distortions were able to establish inter-helical interactions with more number of transmembrane segments in the helical bundle. The study on helix perturbations presented in the previous chapter, brought to light a previously unreported 19 amino acid π-helix fragment interspersed between α-helices in the functionally important transmembrane helix 2 (TM2) belonging to Mitochondrial cytochrome-c-oxidase (1v55). Chapter 6 describes a case study of the structurally similar but functionally different members within the Heme-Copper- Superoxidases (HCO) superfamily that were considered for a comparative analysis of TM2. An analysis of 7 family members revealed that the π-helix shortens, fragments in two shorter π-helices or was even absent in some family members. The long π-helix significantly decreased the total twist and rise of the entire helical fragment thus accommodating more hydrophobic amino acids within the bilayer to avoid hydrophobic mismatch with the bilayer. The increased radius of the TM2 helical fragment also assisted in helix packing interactions by increasing the number of residues involved in helix-helix interactions and hydrogen bonds. Chapter 7 documents the conclusions from the different analyses presented in each of the above chapters. Overall, it is found that membrane proteins optimize the biophysical and chemical constraints of the external environment to strategically place select amino acids at helix termini to ‘start’ and ‘stop’ α-helices. The stabilization of these helix termini is a consequence of sequence dependent structural preferences to form helix capping motifs. The studies on helix transitions and distortions highlight that membrane proteins are not only packed as α-helices but also accomodate 310- and π-helical fragments. These transitions and distortions help in harboring more hydrophobic amino acids and aiding inter-helical interactions important for maintaining the fold of the membrane protein. Appendix A describes a comparison of α-helix assignments in globular and membrane proteins by two algorithms, one based on Cα trace (ASSP) and the other using a combination of hydrogen bond pattern along with backbone torsion angles φ and ψ (STRIDE).

Amorphous silica membranes have molecular sieving properties for the separation of hydrogen from gas mixtures at high temperature. Consequently, they are considered to be applied in separation of a shifted syngas coming out of a water-gas-shift-reactor into the syngas and hydrogen. This separation is a key to an Integrated Gasification Combined Cycle (IGCC) plant, which would allow reducing the carbon footprint in power generation industry. The main objective of this thesis was to carry out a preliminary assessment of suitability of currently available amorphous silica membranes for this separation. However, the separation properties of amorphous silica membranes reported in the open literature vary by orders of magnitude. Therefore, in the first part of this thesis the separation properties of hypothetical silica membrane with different pore size distributions were predicted from first principles. Considering different possible gas transport mechanisms, it was concluded that gas transport in amorphous silica membranes is dominated by the activated and non-activated Knudsen diffusion. The activation energy for transport of different species was predicted using the concept of suction energy. Then, with arbitrary pore size distributions gas permeance of hypothetical silica membrane was predicted for different gas species. Since the pore size distribution of amorphous silica membrane cannot be known a priori, the developed model was used to determine the pore size distribution based on experimentally measured single gas permeances of three different species (kindly provided by Natural Resources Canada, CANMET Energy Technology Center (CETC) laboratory in Ottawa) by minimizing the error of the calculated permeance ratios with respect to the experimental values. The results indicate that, depending on how the objective function is defined, more than one pore size distribution can be found to satisfy the experimental permeance ratios. It is speculated that by increasing the number of experimentally determined permeances, a more unique pore size distribution for the tested silica membrane can be obtained. However, even at this early stage, the developed model provides a rational explanation for the effect of membrane densification on the properties of silica membranes. More specifically, a simultaneous decrease in membrane permeance and selectivity due to membrane densification, reported in the literature, is explained by shrinking the size of pores beyond a certain critical value, which depends on the kinetic diameter of gas molecules that are being separated. Comparing theoretically determined permeances, which match experimentally observed permeance ratios, revealed that the experimental permeances are considerably smaller than the theoretical values. The ratio of the two provided the basis for a scaling factor, a new concept that was introduced in this thesis. To simulate membrane module performance, a novel approach was introduced. More specifically, co- and counter-current flow configurations as well as cross-flow configuration were modeled by assuming no change in feed composition over an infinitesimally small element of membrane area. This led to a system of linear, rather than differential equations, which was readily solved numerically.