A system was developed to study membrane transport in isolated human fibroblast plasma membrane vesicles, avoiding potential complications of intracellular binding and metabolism in the intact cell. Ten-fold enrichment of plasma membranes after subcellular fractionation was confirmed with appropriate markers. Transport competence was established by the following criteria: osmotic sensitivity, stereospecificity, temperature dependence, sodium gradient stimulation, response to ions and ionophores, saturability, and exchange properties. Methotrexate uptake was osmotically sensitive, temperature sensitive, saturable, and inhibited by folinic acid and phosphate. Measurement of lysine uptake was complicated by binding and lack of sodium dependence, but was stimulated by an interior-negative membrane potential, and intravesicular lysine or arginine. Exchange properties of the lysine carrier were exploited to assess its function in fibroblasts from patients with the Mendelian phenotype, lysinuric protein intolerance (LPI). LPI vesicles were not different from controls in their lysine transport phenotype.

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This study assesses the geological constraints on fluid flow within the main crater of Whakaari volcano (White Island) which is located in the Bay of Plenty, New Zealand. A review of the volcanological and morphological history, field mapping, and permeability experiments were used to propose a model for single-state (gas or liquid water) fluid flow in the volcano. Three structural scales were of most importance: (a) the elongate main crater (1.2 km by 0.5 km); which contains (b) three subcraters (~300-500 m in diameter); and (c) >14 historic eruption craters and crater complexes (30-300 m in diameter). A large (2.1x10⁸ m³) sector collapse formed the basic morphology and structure of the amphitheatre-like main crater ≤3.4 ka. Hot fluids are released from magma at ~1–2 km depth and circulated within a conduit-hosted volcano-hydrothermal system. The collapse event was likely to have removed low permeability cone lavas, significantly increasing meteoric water collection and lateral seawater infiltration within high permeability main crater fill above the magma conduit. It is proposed that this caused a susceptibility to ‘wet’ (i.e. phreatic and phreatomagmatic) eruptions which possibly formed three prehistoric subcraters and has been demonstrated in the last ~200 years of available historic record. The permeability of the remaining in-situ cone lavas is controlled by micro- (<1 mm) and macro- (>1 mm) cracks but despite these cracks, the cone lavas’ permeability is still sufficiently low to focus rising magmatic fluid flow through main crater fill. Low-to-high permeability lithified tuffs are inferred to fill the main crater at depth. Low permeability fine ash tuffs generally restrict vertical fluid flow put permit it when vertical trains of vesicles are present. Atmospheric steam and gas pluming is accommodated by a permeable zone of repeated and overlapping historic eruption crater-related discontinuities that extend to >250 m depth through highly permeable unlithified main crater fill in the west. It is likely to be this material into which the seawater infiltrates from the east. Throughout the main crater, fluid flow is focussed at subcrater margins due to steeply-dipping discontinuities between low permeability lava and low-to-high permeability crater fill deposits. The variable permeabilities of crater fill deposits are due to age-related factors of hydrothermal alteration, reworking/sorting, consolidation, and pore mineralisation. At shallow levels (<100 m depth), vertical fluid flow is diverted to historic eruption crater margins by very low permeability clay (reworked and altered tephra). High permeability coarse ash tuffs, Fe-rich lapilli tuffs, and surficial solfatara deposits do not appear to have much effect on the overall fluid flow system. The results of this study show that, within active volcanic craters, the spatial distributions of variably permeable lithologies are often related to discontinuous cratering structures. Together, these are significant geological constraints on fluid flow. Morphological changes to crater structure can directly impact the groundwater regime above the magma conduit and may strongly influence the occurrence of wet versus dry eruptions. This process is possibly a significant control on eruptive behaviour at volcanoes with similar fluid flow systems worldwide.

Azithromycin, an azalide and member of the macrolide group, is a broad spectrum antimicrobial, representing one of the bestselling antimicrobials worldwide. It is derived from erythromycin and exhibits improved acidic stability as a result of its structural modifications. The stable solid form of azithromycin is its dihydrate, although it also naturally occurs in its metastable forms, i.e. the monohydrate and anhydrate. Because azithromycin is poorly soluble in water, its absorption from the gastro-intestinal tract is negatively influenced, which ultimately affects its bioavailability following oral administration (37 %). Polymorphic (monohydrates and dihydrates) and anhydrous forms of azithromycin were screened and investigated. One anhydrous form also proved to be amorphous, which shifted the focus of this study from polymorphism to amorphism. An amorphous glassy azithromycin was subsequently prepared and fully characterised to present its solid state profile. The stability of this amorphous glassy form was established at a high temperature and relative humidity over a period of four weeks. Exposure to increased relative humidity (up to 95 %) and increased water content (up to 50 %) also served as stability indicating tests. Its solubility in various aqueous media was determined. A solid dosage form (tablet), containing the azithromycin glass, was prepared, whereafter these tablets were subjected to dissolution studies in different aqueous media. The stability of azithromycin glass in tablet form was determined over a period of three months. The permeability of azithromycin glass across excised pig intestinal tissue was further established at various pH values. This amorphous glassy form of azithromycin (AZM-G) proved to be very stable at high temperature and relative humidity, whilst also remaining stable after prolonged exposure to 95 % of relative humidity, as it only adsorbed moisture onto its surface. Water content (up to 50 %) had no plasticising effect on azithromycin glass. It demonstrated a significantly higher water solubility (339 % improvement) in comparison with the commercially available azithromycin dihydrate and was it also 39 % more soluble in phosphate buffer (pH 6.8) than its dihydrate counterpart. The prepared azithromycin glass tablets showed a promising dissolution profile in water, due to the improved water solubility of this glass form. The transport of azithromycin glass at higher pH values (6.8 and 7.2) across the membrane proved to be significantly higher than that of azithromycin dihydrate, thus also illustrating its pH dependence for its transport across pig intestinal tissue. The improved water solubility of the azithromycin glass, together with its faster dissolution rate, its superior stability and its increased permeability, may ultimately result in a higher azithromycin bioavailability following oral administration. These research outcomes hence give rise to the need for investigating the effect of administering lower dosages of azithromycin and to determine whether the same antimicrobial efficacy would possibly be achieved, due to maintaining the same tissue concentration levels at these lower dosages. / Thesis (PhD (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013

Azithromycin, an azalide and member of the macrolide group, is a broad spectrum antimicrobial, representing one of the bestselling antimicrobials worldwide. It is derived from erythromycin and exhibits improved acidic stability as a result of its structural modifications. The stable solid form of azithromycin is its dihydrate, although it also naturally occurs in its metastable forms, i.e. the monohydrate and anhydrate. Because azithromycin is poorly soluble in water, its absorption from the gastro-intestinal tract is negatively influenced, which ultimately affects its bioavailability following oral administration (37 %). Polymorphic (monohydrates and dihydrates) and anhydrous forms of azithromycin were screened and investigated. One anhydrous form also proved to be amorphous, which shifted the focus of this study from polymorphism to amorphism. An amorphous glassy azithromycin was subsequently prepared and fully characterised to present its solid state profile. The stability of this amorphous glassy form was established at a high temperature and relative humidity over a period of four weeks. Exposure to increased relative humidity (up to 95 %) and increased water content (up to 50 %) also served as stability indicating tests. Its solubility in various aqueous media was determined. A solid dosage form (tablet), containing the azithromycin glass, was prepared, whereafter these tablets were subjected to dissolution studies in different aqueous media. The stability of azithromycin glass in tablet form was determined over a period of three months. The permeability of azithromycin glass across excised pig intestinal tissue was further established at various pH values. This amorphous glassy form of azithromycin (AZM-G) proved to be very stable at high temperature and relative humidity, whilst also remaining stable after prolonged exposure to 95 % of relative humidity, as it only adsorbed moisture onto its surface. Water content (up to 50 %) had no plasticising effect on azithromycin glass. It demonstrated a significantly higher water solubility (339 % improvement) in comparison with the commercially available azithromycin dihydrate and was it also 39 % more soluble in phosphate buffer (pH 6.8) than its dihydrate counterpart. The prepared azithromycin glass tablets showed a promising dissolution profile in water, due to the improved water solubility of this glass form. The transport of azithromycin glass at higher pH values (6.8 and 7.2) across the membrane proved to be significantly higher than that of azithromycin dihydrate, thus also illustrating its pH dependence for its transport across pig intestinal tissue. The improved water solubility of the azithromycin glass, together with its faster dissolution rate, its superior stability and its increased permeability, may ultimately result in a higher azithromycin bioavailability following oral administration. These research outcomes hence give rise to the need for investigating the effect of administering lower dosages of azithromycin and to determine whether the same antimicrobial efficacy would possibly be achieved, due to maintaining the same tissue concentration levels at these lower dosages. / Thesis (PhD (Pharmaceutics))--North-West University, Potchefstroom Campus, 2013

Cracks in reinforced concrete structures can occur as a result of many phenomena such as fresh concrete bleeding, restrained shrinkage, thermal gradients, freeze-thaw cycles, alkali-aggregate reactions, and can also be induced by external loading. Thus, concrete becomes more vulnerable to the processes of deterioration by corrosion of reinforcement. The corrosion rate of cracked reinforced concrete in different exposure conditions has been studied by some researchers. However, it is not clear how the presence of cracks affects the corrosion-determining factors, which control the corrosion pattern at the crack. The objective of this project was to develop an understanding of the effects of cracking on the transport characteristics under wetting and drying cycles. In this project, flexural loading induced natural cracks, and parallel-wall artificial cracks were studied. The infiltration properties of those cracks were evaluated by the tension infiltrometry technique. The saturation conditions around the crack were monitored with the Time Domain Reflectometry (TDR) technique. A numerical simulation was carried out to model the evolution of saturation in the cracked beams; in the model two crack modeling approaches were employed and compared. The infiltration test showed that the presence of both artificial and natural cracks (0.3 mm and 1.0 mm) dramatically increased the permeability of concrete. The value of hydraulic conductivity was increased by up to 5 orders of magnitude at the location of the crack. The evolution of water saturation of the cracked concrete under wetting and drying conditions was analyzed as colour-scaled images and the water saturation contours were compared for different crack openings. For the artificial crack samples, a deviation from the expected “perfectly symmetric” flow regime around a straight crack was observed. This was probably caused by the micro cracks induced during the shim pull-out process or a non-uniform compaction around the shim insertion. For the natural cracks, in the drying phase, smaller cracks seemed to have better water storage. Hence, the water saturation decreased at a slightly slower rate. The crack behaved like an open surface that was exposed to the environment. Application of the same material properties to the open surface and the crack surface did not bring a large error for the water flow simulations. A hysteresis phenomenon has been found during the identification of the Van Genuchten material parameters using an inverse modelling approach, with Ks=5×10-10 m/s, α =4.33×10-4, for the wetting phase, n=1.32 and for the drying phase, n=2.0. The simulation results suggest that for the simple flexural crack, the 1D crack line averaged from the front and back crack lines is capable of representing the crack in the wetting and drying scenario. The crack could be modelled as “free surface” or “equivalent porous medium”.