>Magister Scientiae - MSc / 68Ge/68Ga generators rely on metal oxide, inorganic and organic sorbents in order to prepare radionuclides useful for clinical applications. The requirements for 68Ge/68Ga generators are that the 68Ga obtained from the 68Ge loaded column should be optimally suited for the routine synthesis of 68Ga-labelled radiopharmaceuticals, that the separation of the 68Ga daughter from the 68Ge parent should happen easily, with a high yield of separation, a low specific volume of 68Ga and should not contain trace elements owing to the solubility of the metal oxide sorbent. Beginning with a metal oxide preparation and continuing through recent developments, several approaches for processing generator derived 68Ga have altered the production of 68Ge/68Ga generators. Still, the effects of sorbent modification on the properties of 68Ge/68Ga radionuclide generator systems are not necessarily optimally designed for direct application in a medical context. The objective of this research was to analyze and document characteristics of Titanium Oxide (TiO2) sorbents relevant to processing of a 68Ge/68Ga generator that is able to produce 68Ga eluates that are adequate for clinical requirements. Interest was shown in TiO2 based 68Ge/68Ga generators by a number of overseas companies for tumour imaging using 68Ga-labelled 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-conjugated peptides. While a method involving production of the 68Ga radionuclide using TiO2 metal oxide had been published, problems with the production persisted. A method, using TiO2 metal oxide for ion exchange chromatography, was devised in this study to produce the 68Ga radionuclide, with the aim of being adopted for production purposes. The study focuses on the development of a dedicated procedure for the achievement of sufficient 68Ga yield along with low 68Ge breakthrough and low metallic impurities. Literature from 1970 to 2011 was reviewed to assess the radiochemical aspects of the 68Ga production and processing thereof. Various commercially available TiO2 metal oxides were characterized by subjecting the materials to x-ray diffraction (XRD), x-ray fluorescence (XRF) and scanning electron microscopy (SEM) for quantitative and qualitative analysis.

Magister Scientiae (Medical Bioscience) - MSc(MBS) / The blood brain barrier (BBB) is a signaling interface between the blood and the central nervous system (CNS), which prohibits the entry of harmful blood-borne substances into the brain micro-environment, thus maintaining brain homeostasis. The crucial role of the BBB is protecting the CNS, which may adversely be affected by alcohol. The central component of the BBB, endothelial cells (ECs), regulates BBB transport by regulating the permeability both transcellularly and through their paracellular junctions, by structures called tight junctions (TJs) that are composed of proteins. The aim of this study was to investigate the in vitro effects of ethanol (EtOH) and fermented rooibos (Rf) on a monolayer of bEnd5 mouse brain ECs, by determining the effects of EtOH and Rf on bEnd5 (i) cell viability (ii) cell proliferation (iii) rate of cell division (iv) cell toxicity (v) claudin-5 transcription (vi) permeability across a monolayer of bEnd5 ECs and (vii) morphology, for a selected experimental timeline of 24, 48, 72, and 96hrs. We then investigated if the simultaneous exposure of Rf and EtOH could reverse or alleviate the EtOHinduced effects on the bEnd5 ECs. EtOH metabolism induces oxidative stress and results in a range of adverse physiological effects. Aspathalus linearis (rooibos) contains many phenolic compounds, of which the main antioxidant activity is attributed to aspalathin. Our underlining hypothesis is that the antioxidants in an aqueous rooibos extract may therefore protect against the potential oxidant damaging effects of alcohol on the BBB. Cells were exposed for 24hrs to selected concentrations of EtOH (25mM and 100mM), a concentration of Rf containing equivalent of 1.9nM aspalathin, and the combinations of EtOH and Rf. Cell viability and cell toxicity was determined, while cell proliferation and rate of cell division was estimated using the trypan blue exclusion assay. Real time quantitative PCR was implemented to quantify claudin-5 transcription, normalized against housekeeping genes, GAPDH and HPRT. Transepithelial electrical resistance (TEER) was measured using the Ohm Millicell-electrical resistance system, while bEnd5 monolayer morphology was analysed using the Zeiss scanning electron microscope. Both concentrations of EtOH led to an overall decrease in cell viability, and a decreased number of live cells across 72hrs. Consistent with this, EtOH resulted in increased cell toxicity across the 96hr experimental timeframe and a diminished rate of cell division. The transcription of claudin-5 in bEnd5 ECs exposed to 25mM and 100mM EtOH varied dramatically across the 96hr timeframe. While 25mM EtOH resulted in an overall decrease in TEER, cells exposed to 100mM EtOH only decreased TEER between 48 and 96hrs. Morphologically, both concentrations of EtOH led to compromised paracellular spaces as endorsed by high definition SEM analysis. The administration of Rf on its own resulted in an initial decrease in viability, followed by recovery between 72 and 96hrs. Exposure to Rf diminished live cell numbers at 72 and 96hrs, accompanied by a compromised rate of cell division and an overall increase in cell toxicity. In addition, Rf down-regulated claudin-5 transcription across the course of the experiment, particularly between 24 and 48hrs. In alignment with this, Rf also led to an increase in BBB permeability from 24 to 96hrs. However, SEM studies were not able to discriminate any differences between control and Rf treated cells. Our study showed that the BBB could be protected against the adverse effects of EtOH, and this at the plasma concentration induced by 500ml’s of Rooibos tea. The simultaneous exposure of Rf and EtOH was able to negate the effects of EtOH on cell viability, cell proliferation, and cell toxicity but exacerbated the effects of EtOH on claudin-5 transcription and paracellular permeability. Morphologically, co-exposure with Rf only reversed the effects of 25mM EtOH while exacerbating the effects of 100mM EtOH at 96hrs. In conclusion, EtOH was shown to be detrimental to the integrity of bEnd5 ECs, and the addition of a minuscule quantity of t h e Rf extract was able to partially alleviate excess ROS-induced effects. / National Research Foundation (NRF)

>Magister Scientiae - MSc / Efavirenz is a non-nucleoside reverse transcriptase inhibitor used as an anti-retroviral for the treatment of human immunodeficiency virus (HIV) type I. It is classified as a class IΙ drug under the Biopharmaceutical Classification System (BCS) and exhibits a low solubility (aqueous solubility of 9.0 μg/ml) and high permeability (variable oral bioavailability). This study aims to choose a pre-formulation protocol with the best efavirenz derivative in literature between co-crystals and CD inclusion complexes. Upon selection of the efavirenz derivative, the complications of both small scale and large scale laboratory pre-formulation production is highlighted for formulation of a tablet dosage form. Numerous variables were selected for the pre-formulation protocol. Physical, chemical, pharmacological, pharmaceutical and economical variables were investigated. Citric acid monohydrate (CTRC) was chosen as the best co-former for a co-crystal while hydroxypropyl-beta-cyclodextrin (HP-β-CD) was selected as a host for an inclusion complex. Pharmaceutically, the angle of repose, Carr’s index, Hausner’s ratio, moisture content, disintegration time, hardness/resistance to crush, manufacturing process problems and particle size of the CTRC and HP-β-CD were all evaluated. The CTRC was ultimately selected for formulation of a tablet. The preparation of small laboratory scale of EFA/CTRC co-crystal was successfully achieved after several attempts. The large laboratory scale of EFA/CTRC was prepared under various environmental seasons which were indicated as batches 1-6 for purposes of this study. Characterization of the large laboratory scale EFA/CTRC co-crystals was performed by scanning electron microscopy (SEM), hot-stage microscopy (HSM), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and by physical inspection (i.e. season, texture, colour, shape and particle size) of the EFA/CTRC product. Batch 1 and 2 were prepared during the summer season. The SEM analysis showed that the particles were needle-like shaped. The thermal analysis values of batch 1 by HSM, DSC and TGA results were 123 °C, 119 °C and 1.68 % of mass loss, respectively. In batch 2, morphology results by SEM revealed spikes of irregular and agglomerated particles. Batch 2 melted at 123 °C and a small unmelted quantity was observed at 143 °C. The DSC and TGA (mass loss) analysis were 118 °C and 0.75 % respectively. The hardness test of EFA/CTRC tablet prepared in batch 2 was extremely hard hence failed the disintegration test. The EFA/CTRC prepared in batches 3, 4 and 5 was during the winter season which is associated with high humidity and wet weather conditions. The SEM, DSC, TGA results were significantly different from the previous batches. The SEM morphology was highly irregular particles for batch 3, clustered and randomly size particle for batch 4 and irregular, needle-like, spikes and spherical shaped particles for batch 5, respectively. The thermal results HSM, DSC and TGA confirmed the presence of moisture in the prepared EFA/CTRC products. The HSM melting point results of batches 3, 4 and 5 were 123 °C, 115 °C and 121 °C, respectively. The DSC results of 110 °C, 105 °C and 118 °C were observed for batches 3, 4 and 5 respectively. The mass loss i.e. TGA results for batches 3, 4 and 5 were 1.178%, 1.5 % and 2.235 % respectively. In batch 6, EFA/CTRC was prepared using a different commercial batch of EFA and CTRC. The SEM results indicated the formation of needle-like and clustered particles. The values obtained from HSM, DSC and TGA results were 124 °C, 114 °C and 0.54 % in mass loss. The physical appearance of EFA/CTRC prepared from batch 1 and 2 were white in colour while batch 3, 4, 5 and 6 of the prepared EFA/CTRC was pink in colour. The physical appearance of the individual batches differed but the identity of the sample remained intact implying the same pharmacological effects with differing pharmaceutical properties impacting the dosage form preparation.

Doctor Scientiae / Ti – based AB and AB₂ hydride forming materials have shown to be very promising hydrogen storage alloys due to their reasonable reversible hydrogen storage capacity at near ambient conditions, abundance and low cost. However, these materials are not used extensively due to their poor activation performances and poisoning tolerance, resulting insignificant impeding of hydrogen sorption. The overall goal of this project was to develop the knowledge base for solid-state hydrogen storage technology suitable for stationary and special vehicular applications focussing mainly on Ti – based metal hydrides. In order to accomplish this goal, the project had a dual focus which included the synthesis methodology of Ti – based AB and AB₂ materials and the development of new surface engineering solutions, based on electroless plating and chemical vapour deposition on the surface modification of Ti – based metal hydride forming materials using Pd-based catalytic layers. TiFe alloy was synthesised by sintering of the Ti and Fe powders and by arc-melting. Sintered samples revealed three phases: TiFe (major), Ti₄Fe₂O, and β-Ti. Hydrogen absorption showed that the sintered material was almost fully activated after the first vacuum heating (400 °C) when compared to the arc-melted sample requiring several activation cycles. The increase in the hydrogen absorption kinetics of the sintered sample was associated with the influence of the formed hydrogen transfer catalyst, viz. oxygen containing Ti₄Fe₂O₁₋ₓ and β-Ti, which was confirmed by the XRD data from the samples before and after hydrogenation. The introduction of oxygen impurity into TiFe alloy observed in the sintered sample significantly influenced on its PCT performances, due to formation of stable hydrides of the impurity phases, as well as destabilisation of both β-TiFeH and, especially, γ-TiFeH₂. This finally resulted in the decrease of the reversible hydrogen storage capacity of the oxygen-contaminated sample. TiFe alloy was also prepared via induction melting using graphite and alumo-silica crucibles. It was shown that the samples prepared via the graphite crucible produced TiFe alloy as the major phase, whereas the alumo-silica crucible produced Ti₄Fe₂O₁-x and TiFe₂ as the major phases, and TiFe alloy as the minor one. A new method for the production of TiFe – based materials by two-stage reduction of ilmenite (FeTiO₃) using H₂ and CaH₂ as reducing agents was developed. The reversible hydrogen absorption performance of the TiFe – based material prepared via reduction of ilmenite was 0.5 wt. % H, although hydrogen absorption capacity of TiFe reported in the literature should be about 1.8 wt. %. The main reason for this low hydrogen capacity is due to large amount of oxygen present in the as prepared TiFe alloy. Thus to improve the hydrogen absorption of the raw TiFe alloy, it was melted with Zr, Cr, Mn, Ni and Cu to yield an AB₂ alloy. For the as prepared AB₂ alloy, the reversible hydrogen sorption capacity was about 1.3 wt. % H at P=40 bar and >1.8 wt.% at P=150 bar, which is acceptable for stationary applications. Finally, the material was found to be superior as compared to known AB₂-type alloys, as regards to its poisoning tolerance: 10-minutes long exposure of the dehydrogenated material to air results in a slight decrease of the hydrogen absorption capacity, but almost does not reduce the rate of the hydrogenation. Hydrogen storage performance of the TiFe-based materials suffers from difficulties with hydrogenation and sensitivity towards impurities in hydrogen gas, reducing hydrogen uptake rates and decreasing the cycle stability. An efficient solution to this problem is in modification of the material surface by the deposition of metals (including Palladium) capable of catalysing the dissociative chemisorption of hydrogen molecules. In this work, the surface modification of TiFe alloy was performed using autocatalytic deposition using PdCl₂ as the Pd precursor and metal-organic chemical vapour deposition technique (MO CVD), by thermal decomposition of palladium (II) acetylacetonate (Pd[acac]₂) mixed with the powder of the parent alloy. After surface modification of TiFe – based metal hydride materials with Pd, the alloy activation performance improved resulting in the alloy absorbing hydrogen without any activation process. The material also showed to absorb hydrogen after exposure to air, which otherwise proved detrimental.

>Magister Scientiae - MSc / Polymer electrolyte membrane fuel cells (PEMFCs) among other fuel cells are considered the best candidate for commercialization of portable and transportation applications because of their high energy conversion and low pollutant emission. Recently, there has been significant interest in high temperature polymer electrolyte membrane fuel cells (HT-PEMFCs), due to certain advantages such as simplified system and better tolerance to CO poisoning. Cost, durability and the reliability are delaying the commercialization of PEM fuel cell technology. Above all durability is the most critical issue and it influences the other two issues. The main objective of this work is to study the durability of membrane electrode assemblies (MEAs) for HT-PEMFC. In this study the investigation of commercial MEAs was done by evaluating their performance through polarization studies on a single cell, including using pure hydrogen and hydrogen containing various concentrations of CO as fuel, and to study the performance of the MEAs at various operating temperatures. The durability of the MEAs was evaluated by carrying out long term studies with a fixed load, temperature cycling and open circuit voltage degradation. Among the parameters studied, significant loss in the performance of the MEAs was noted during temperature cycling. The effect of temperature cycling on the performance of the cell showed that the performance decreases with increasing no. of cycles. This could be due to leaching of acid from the cell or loss of electrochemically active surface area caused by Pt particle size growth. For example at 160°C, a performance loss of 3.5% was obtained after the first cycle, but after the fourth cycle a huge loss of 80.8% was obtained. The in-house MEAs with Pt-based binary catalysts as anodes were studied for CO tolerance, performance and durability. A comparison of polarization curves between commercial and in-house MEAs illustrated that commercial MEA gave better performance, obtaining 0.52 A/cm² at 0.5V and temperature of 160°C, with in-house giving 0.39A/cm² using same parameters as commercial. The CO tolerance of both commercial and in-house MEA was found to be similar. In order to increase the CO tolerance of the in-house MEAs, Pt based binary catalysts were employed as anodesand the performance was investigated In-house MEAs with Pt/C and Pt-based binary catalysts were compared and a better performance was observed for Pt/C than Pt-alloy catalysts with Pt-Co/C showing comparable performance. At 0.5 V the performance obtained was 0.39 A/cm2 for Pt/C, and 0.34A/cm²,0.28A/cm²,0.27A/cm² and 0.16A/cm² were obtained for Pt-Co/C, Pt-Fe/C, Pt-Cu/C and Pt-Ni respectively. When the binary catalysts were tested for CO tolerance, Pt-Co showed no significant loss in performance when hydrogen containing CO was used as anode fuel. Scanning electron microscopy (SEM) revealed delamination between the electrodes and membrane of the tested and untested MEA's. Membrane thinning was noted and carbon corrosion was observed from the tested micro-porous layer between the gas diffusion layer (GDL) and catalyst layer (CL).

Estudos morfológicos representam ferramentas básicas para ampliação do conhecimento sobre determinado táxon e abrem caminhos para o desenvolvimento de outros estudos em diversas áreas como taxonomia, filogenia, etologia e ecologia. Neste trabalho, buscamos aprimorar o conhecimento morfológico na família Pentatomidae, investigando as variações presentes nas pernas de percevejos e como elas se desenvolvem ao longo do ciclo de vida. Estudando comparativamente as expansões tibiais e o aparato tibial na subfamília Asopinae, e verificamos que ambas as estruturas possuem uma ampla variação morfológica. As expansões podem estar presentes nas superfícies dorsal e ventral da tíbia, ou apenas na dorsal. A estrutura pode apresentar diferentes tamanhos entre as espécies, podendo ser até duas vezes a largura do eixo central da tíbia como nos gêneros Cazira e Heteroscelis, ou em outros casos como Alceorrhynchus, que se apresenta de forma diminuta. Além disso, as expansões podem ocorrer ao longo de toda a tíbia, ou apenas apicalmente. Em relação ao aparato tibial, verificamos que o número de cerdas é bastante variável entre as espécies analisadas, e que possui relação com o tamanho geral do inseto. Ainda, verificamos que a região circundante à estrutura apresenta morfologia variável. Com base nas variações encontradas, dez caracteres morfológicos foram propostos. Além dos adultos, estudamos a morfologia e alometria da tíbia ao longo da ontogenia em nove espécies de Pentatomidae das subfamílias Asopinae, Pentatominae e Edessinae. Foi constatado que o número de cerdas aumenta em cada estágio de desenvolvimento nas espécies de Asopinae, enquanto nas demais o número se mantém praticamente o mesmo. A alometria do aparato tibial se demonstrou positiva para todas as espécies de Asopinae nos testes realizados, enquanto nas subfamílias Pentatominae e Edessinae a alometria da estrutura foi maioritariamente negativa ou isométrica. Foi demonstrado que assim como em outros insetos, as pernas apresentam um crescimento acelerado em comparação com outras estruturas do corpo, neste caso, da cabeça. Além disso, constatamos que a largura de tíbia apresenta uma queda de tamanho do quinto ínstar para o estágio Adulto, e hipotetizamos que este fenômeno possa ter relação com a perda de estruturas presentes nas pernas das ninfas responsáveis por dar maior suporte ao peso do inseto. Os estágios imaturos de Tynacantha marginata Dallas são descritos, estruturas corporais são exploradas e comparadas morfologicamente, e características diagnósticas que permitem a identificação dos ovos e ninfas da espécie são fornecidas. / Morphological studies are basic tools to increase knowledge about a given taxon, opening paths for the development of other studies in several areas such as taxonomy, phylogeny, ethology and ecology. In this work, we aimed to improve the morphological knowledge in the Pentatomidae family, examining the variations present in the legs of stink bugs and how they develop throughout the life-cycle. Comparatively studying the tibial expansions and the foretibial apparatus in the Asopinae subfamily, we found that both structures have a wide morphological variation. The expansions may be present on the dorsal and ventral surfaces of the foretibia, or only on the dorsal surface. The structure can present different sizes between the species, being able to be up to twice the width of the tibial axis as in the genus Cazira and Heteroscelis, or in other cases like Alceorrhynchus, appears in a diminutive form. Besides, the expansions may occur along the entire tibia, or only apically. In relation to the foretibial apparatus, we verified that the number of setae is quite variable among the analyzed species, and that it is related with the general size of the insect. Also, we verified that the region surrounding the structure presents variable morphology. Based on the variations found, ten morphological characters are proposed. In addition to the adults, we studied the morphology and allometry of the foretibia along the ontogeny in nine species of Pentatomidae of the subfamilies Asopinae, Pentatominae and Edessinae. It was observed that the number of setae increases at each stage of development in the Asopinae species, while in the others the number remains practically the same. Allometry of the tibial apparatus was shown to be positive for all Asopinae species in the performed tests, while in the Pentatominae and Edessinae subfamilies the allometry of the structure was mostly negative or isometric. It has demonstrated that, as in other insects, the legs exhibit accelerated growth compared to other structures of the body, in our case, the head. In addition, we verified that tibia width shows a decrease in size of the fifth instar for the adult stage, and hypothesized that this phenomenon is related to the loss of structures present in the legs of the nymphs responsible for giving support to the weight of the insect. The immature stages of Tynacantha marginata Dallas are described, body structures are explored and morphologically compared and diagnostic features that allow identification of the eggs and nymphs of the species are provided.

Magister Pharmaceuticae - MPharm / The aim of this study was to investigate the mechanism of fenfluramine-induced pulmonary phospholipidosis, by comparing the profile and levels of induced phospholipids in the rat and the mode of phospholipase inactivation, both relative to that produced by chlorphentermine. Wistar and BD9 rats were injected with fenfluramine (FF) and chlorphentermine (CP) intra-peritoneally daily over a six week period to induce phospholipidosis. The lungs isolated from such treated and untreated animals, were grouped into unlavaged lungs and lungs to be lavaged and from the latter group the alveolar macrophages were isolated. Small sections of the unlavaged lungs were microscopically examined to verify the induction of phospholipidosis. Further the levels of phosphatidyl choline (PC), spingomyelin (SPM), phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), phosphatidyl inositol (PI), phosphatidyl serine (PS) and phosphatidic acid (PA) were determined in both groups of lungs using a TLC method. To assess whether the drug-mediated inactivation of the phospholipases (PL) occurred via direct inhibition of the enzymes or via the drug-phospholipid complex, the hydrolysis of the above phospholipids by PL-A or PL-C were monitored using colorimetric methods. The feasibility of the phospholipid-drug complex-mediated mechanism was further explored, by assessing the effect the two drugs had on the phase transition temperature of the phospholipids. Electron microscopy revealed the presence of hypertrophied and elevated counts of alveolar macrophages in the treated-Wistar and -BD9 rats. In the FF- and CP treated Wistar and BD9 rats there were, compared to the saline-treated rats, a 200 % and 235 % increase in macrophage counts, respectively, for the FF-treated rats and a 700 % and 965 % increase in macrophage counts, respectively, for the CP treated rats. The levels of all the phospholipids in the unlavaged lungs of both rat strains were elevated, except that for PG, PS and PA. In both rat strains following the treatment with both drugs the PG levels were not elevated and the PS levels were not elevated following CP treatment. Following the treatment for both drugs, the PA levels were also not elevated in the BD9 rats. Relative to the levels found in the unlavaged lungs of the control rats, the increases ranged from a minimum of 9 to a maximum of 216 %. In general, Wistar rats appeared to be more susceptible to both FF and CP treatment. In both rat strains, lavaging of the lungs considerably reduced the levels of phospholipids remaining in the lung and the differences between the treated and untreated animals became less striking. The addition of FF or CP, whether directly to the enzyme, or in the form of the drug phospholipid complex, resulted in significant decreases in the PL-A-mediated or PL-C-mediated hydrolysis of virtualy all the test phospholipids. The average decrease ranged from 0.811 to 4.04 ,.,.FFAbbb ,.,.1-1sample min-I, for the PL-A activity and 0.023 to 0.827 ,.,.gIp'CC100 ,.,.1-1 sample min-I, for the PL-C activity. In the case of FF, the inhibition of PL-A activity could not be ascribed exclusively to either direct inhibition of the enzyme or reduced susceptibility of the phospholipid substrate-drug complex. The PL-C activity appeared to be inhibited to a greater extent via the phospholipid substrate-drug complex rather than by direct inhibition. On the other hand, CP induced a small, but significantly greater degree of inhibition of PL-A activity, more via direct inhibition, rather than by the phospholipid substrate-drug complex. The PL-C activity appeared to be inhibited to a greater extent via phospholipid substrate-drug complexation than by direct inhibition. From the above data, considered collectively, it was not possible to declare either of the two possible mechanisms as the more likely one for FF or CP-induced inhibition of the phospholipases. The feasibility of the indirect mode was further explored, by determining the phase transition temperatures for the phospholipid-drug complexes of each drug. The addition of each drug caused a depression of the phase transition temperature of all the phospholipids with a .1T'dd ranging from 0.52 to 15.73 °C. This appears to support the notion that both drugs bind to the phospholipids and the differences in the extent of the phase transition temperature depression of the individual phospholipids may indicate differences in the binding capacities of these drugs. The following major conclusions may be drawn from the results of this investigation. Fenfluramine induces a phospholipidosis syndrome in the lungs of Wistar and BD9 rats that are histologically similar to that induced by CP. It induces the elevation of essentially the same phospholipids as CP, primarily in the alveolar spaces and macrophages, and by implication, most likely via similar mechanisms. For both FF and CP, both direct inhibition and phospholipid-drug complex-mediated inhibition of phospholipases were found to be a viable mechanism for this syndrome. The mechanism for FF-induced pulmonary phospholipidosis thus appears to be similar to that of CP; small quantitative differences in essentially similar mechanisms, may explain the differences in the levels of induced phospholipidosis found in this study.

Biofilms of Actinobacillus succinogenes, grown in a biofilm reactor system, were investigated for structure and cell viability, through microscopic visualisation with a confocal scanning laser microscope (CSLM) and a scanning electron microscope (SEM). Biofilms were sampled and visualised at steady state conditions with the broth containing succinic acid titres between 15 and 21 g/L. All sampled biofilm was 6 days old. Six-day-old biofilms of A. succinogenes showed a heterogeneous biofilm architecture composed of cell micro-colony pillars which varied considerably in thickness, area and shape. Microcolony pillars consisted of a densely packed entanglement of sessile cells. Quantitative analysis revealed that the pillars were mostly large, with a mean pillar diameter of 170 m and a mean thickness of 92 m, although pillar diameter and thickness were variable as they ranged from 25 – 500 m and 30 – 300 m, respectively. In the regions close to the substratum surface, pillars were characterised by having defined borders with a network of channels ranging from 40 – 200 m in width separating them. However, towards the middle of the biofilm depth some of the pillars coalesced. For this reason low cross sectional area coverage of biofilm consistently occurred at the bottom portion of the biofilm whilst the highest coverage was in the middle portion of the biofilm. Regarding cell morphology, very large differences were observed. Planktonic cells were rod-shaped, whereas sessile cells expressed an elongated rod morphology and thus were much longer and thinner compared with planktonic cells. Planktonic cells were 1 – 2 m thick and 4 – 5 m long, while sessile cells were 0.5 – 1 m thick and 5 – 100 m long. Long sessile cells resulted in extensive tangling in microcolony pillars, which may have contributed to the structural stability of the pillars. Fibre-like connections of constant diameter were observed between cells, and between the cells and surface. The diameter of these connections was approximately 20 – 30 nm. Viability stains showed that in the bottom portion (from 0 - 20 m above the substratum surface) of the biofilm, most of the cells were dead. However, the portion of covered area attributed to living cells increased past the middle of the biofilm towards the top part of the biofilm. A high percentage of living cells was thus found towards the top part of the biofilm. Overall, 65% (with 2% standard deviation) of the entire biofilm was composed of dead cells. In this way, the results show that operation at high acid conditions comes at a cost of low overall biomass productivity due to decreased active biomass. / Dissertation (MEng)--University of Pretoria, 2017. / Chemical Engineering / MEng / Unrestricted

Despite the significant progress in medicine, infectious diseases are life-threatening thanks to an increasing number of multiresistant strains of microorganisms and late detection of pathological agents. An opportunistic fungus Aspergillus fumigatus cause respiratory system diseases called aspergillosis. The invasive pulmonary aspergillosis affects immunocompromised patients after inhalation of ubiquitous conidia of A. fumigatus and results in 450,000 deaths per year. The biofilm formation in the infected tissue protects A. fumigatus against antimicrobial drugs. Late therapy may not be effective. Infection of immunocompromised patients and biofilm formation is characteristic also for gram negative bacteria Pseudomonas aeruginosa, which is due to the production of many factors of virulence and multiresistance a dreaded opportunistic pathogen. Scanning electron microscopy (SEM) provides detail information about morphology of microorganisms with the resolution in range of tens of nanometers that allows to observe microorganisms in the infected tissue and its pathological changes. Mass spectrometry allows to detect infection and its course based on identification of characteristic microbial molecules. The aim of this study was to optimize sample preparation of tissues infected with A. fumigatus or P....

In this work phase transformations in metastable β (primarily Ti-15Mo) alloys were studied utilizing electrical resistance, dilatometry, transmission electron microscopy and X-ray and neutron diffraction. The materials Ti-15Mo, Ti-6.8Mo-4.5Fe-1.5Al (LCB), Ti-5Al-5V-5Mo-3Cr (Ti-5553), Ti-29Nb-1Fe-0.5Si (TNFS), Ti-15Mo-3Nb-3Al-0.2Si (Timetal 21S) and Ti-13Cr-1Fe-3Al (TCFA) (in wt. %) - were subjected to a solution treatment at a temperature above β transus and quenched into water. In this condition, the microstructure of the investigated materials consists of β matrix and ω particles. Samples quenched from important temperatures determined from in-situ electrical resistance and dilatometry measurements were studied by post-mortem TEM. In-situ X-ray and neutron diffraction provided direct observations of microstructure of Ti-15Mo alloy during linear heating and confirmed statements based on results of indirect methods, such as: the decrease of volume fraction of ω phase during heating at low temperatures (up to 250 ◦ C), complete dissolution of ω phase at 560 ◦ C and precipitation of α phase without ω particles serving as its direct precursors. X-ray diffraction experiment allowed to determine relative evolution of the size of ω particles while phase fraction evolution was derived from neutron diffraction. The...