Call number: LD2668 .T4 1985 S565 / Master of Science

Intracellular transport of membrane-bound vesicles and organelles is a process fundamental for many cellular functions including cell morphogenesis and signaling. The transport is mediated by ensembles of motor proteins, such as kinesins, walking on microtubule tracks. When transporting membrane-bound cargo inside a cell, the motors are linked to diffusive lipid bilayers either directly or via adaptor molecules. The fluidity of the lipid bilayers induces loose inter-motor coupling which is likely to impact the collective motor dynamics and may induce cooperativity. Here, we investigate the influence of loose coupling of kinesin motors on its transport characteristics. In the first part of this thesis, we used truncated kinesin-1 motors with a streptavidin-binding-peptide (SBP) tag and performed gliding motility assays on streptavidin-loaded biotinylated supported lipid bilayers (SLBs), so called ‘membrane-anchored’ gliding motility assays. We show that the membrane-anchored motors act cooperatively; the microtubule gliding velocity increases with increasing motor density. This is in contrast to the transport behavior of multiple motors rigidly bound to a substrate. There, the motility is either insensitive to the motor density or shows negative interference at higher motor density, depending on the structure of the motors. The cooperativity in transport driven by membrane-anchored motors can be explained as following: while stepping on a microtubule, membrane-anchored motors slip backwards in the viscous membrane, thus propelling the microtubule in the solution at a velocity, given by the difference of the motor stepping velocity and the slipping velocity. The motor stepping on the microtubule occurs at maximal stepping velocity because the load on the membrane-anchored motors is minute. Thus, the slipping velocity of membrane-anchored motors determines the microtubule gliding velocity. At steady state, the drag force on the microtubule in the solution is equal to the collective drag force on the membrane-anchored motors slipping in the viscous membrane. As a consequence, at low motor density, membrane-anchored motors slip back faster to balance the drag force of the microtubule in the solution. This results in a microtubule gliding velocity significantly lower than the maximal stepping velocity of the individual motors. In contrast, at high motor density, the microtubules are propelled faster with velocities equal to the maximal stepping velocity of individual motors. Because, in this case, the collective drag force on the motors even at very low slipping velocity, is large enough to balance the microtubule drag in the solution. The theoretical model developed based on this explanation is in good agreement with the experimental data of gliding velocities at different motor densities. The model gives information about the distance that the diffusing motors can isotropically reach to bind to a microtubule, which for membrane-anchored kinesin-1 is ~0.3 µm, an order of magnitude higher as compared to rigidly bound motors, owing to the lateral mobility of motors on the membrane. In addition, the model can be used to predict the number of motors involved in transport of a microtubule based on its gliding velocity. In the second part of the thesis, we investigated the effect of loose inter-motor coupling on the transport behavior of KIF16B, a recently discovered kinesin motor with an inherent lipid-binding domain. Recent studies based on cell biological and cell extract experiments, have postulated that cargo binding of KIF16B is required to activate and dimerize the motor, making it a superprocessive motor. Here, we demonstrate that recombinant full-length KIF16B is a dimer even in the absence of cargo or additional proteins. The KIF16B dimers are active and processive, which demonstrates that the motors are not auto-inhibited in our experiments. Thus, in cells and cell extracts Kif16B may be inhibited by additional factors, which are removed upon cargo binding. Single molecule analysis of KIF16B-GFP reveals that the motors are not superprocessive but exhibit a processivity similar to kinesin-1 indicating that additional factors are most likely necessary to achieve superprocessivity. Transport on membrane-anchored KIF16B motors exhibited a similar cooperative behavior as membrane-anchored kinesin-1 where the microtubule gliding velocity increased with increasing motor density. Taken together, our results demonstrate that the loose coupling of motors via lipid bilayers provides flexibility to cytoskeletal transport systems and induces cooperativity in multi-motor transport. Moreover, our ‘membrane-anchored’ gliding motility assays can be used to study the effects of lipid diffusivity (e.g. the presence of lipid micro-domains and rafts), lipid composition, and adaptor proteins on the collective dynamics of different motors.

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Thesis (MScEng (Mechanical and Mechatronic Engineering))--University of Stellenbosch, 2006. / The quest for higher efficiency of the internal combustion engine will always be pursued. Increasingly stringent emission regulations are forcing manufacturers to downsize on engine displacement and increase specific power. By adding a turbocharger, the airflow throught he engine and hence the specific power can be increased.

Thesis (MScEng (Mechanical and Mechatronic Engineering))--University of Stellenbosch, 2005. / This study involved the modeling and verification of the finger follower valve train arrangement with the focus on the non-linear behaviour of high speed valve springs. The project was divided into three phases namely; the measuring of valve train dynamics, modeling of the valve train and the verification of the dynamic models by comparing the results from the two aforementioned two phases. Acceleration and force were measured on a running engine. A force transducer was specially developed for this purpose. Digital signal processing was used in the analysis and implementation of all measured data. The spring model was developed systematically from a solid model, into a finite element model, and finally into a dynamic model. All development steps were continually checked with experiments and calculations. The primary concept used in the spring modeling lends itself to modal analysis theory in conjunction with the superimposing of non-linearities onto a linear model. The dynamic model was verified and good correlations were found, especially at high engine speeds where valve train dynamics play an important role. Parameter adjustments could be made in the dynamic model and the effect that some engine mechanisms, such as engine oil aeration, had on the valve train dynamics were identified. The project was concluded with a case study of a cam profile optimisation project. Dynamic problems were found that would normally not have been identified without the dynamic model.

Thesis (MScEng)--University of Stellenbosch, 2001. / ENGLISH ABSTRACT: This thesis is concerned with unsteady, one-dimensional flow, which closely mimics those found in the manifolds of internal combustion engines. The physical equations describing problems of this nature are presented and some of the important concepts introduced. These equations and concepts were verified by comparison to published results. The Method of Characteristics (MaC) for unsteady one-dimensional flow with friction and heat transfer was used to analyse the gas flow through the inlet and exhaust systems of an engine. The theoretical derivation of unsteady gas dynamic boundary conditions is presented and the integration with the unsteady pipe flow explained. A simulation flow model was developed to analyse the flow by using the Mae. Thisflow model was then incorporated into an engine simulation program, ESA,to simulate internal combustion engines and to predict the performance of a specific engine. A cam-profile model and an in-cylinder thermodynamic model are used to complete the ESAsoftware. Experimental work was done on a modified Nissan Z24/NA20 engine to evaluate the simulation model. The manifolds of the Nissan Z24/NA20 were modified to isolate one of the cylinders for a proper single cylinder model. More experimental work was done on a Volkswagen 1.6£ 8-valve and a 1.6£ 20-valve engine to obtain performance data on two inlet manifolds developed using the ESAsoftware. Performance data and pressure traces in the inlet manifold of the Nissan Z24/NA20 were recorded for comparison with the ESA software. Good correspondence was found between tested and modelled data and the differences varied between ±5% on engine performance data and pressure wave frequency predictions, and ± 10% on pressure pulse amplitudes. / AFRIKAANSE OPSOMMING: Hierdie tesis handeloor bestendige, eendimensionele vloei, wat die gasvloei in spruitstukke van binnebrandenjins naboots. Die nodige vergelykings wat hierdie tipe probleme beskryf asook van die belangrikste konsepte, word bespreek. Hierdie vergelykings en konsepte is met behulp van gepubliseerde data geverifieer. Die Metode van Karakteristieke (MVK) vir bestendige, eendimensionle vloei met wrywing en warmte oordrag, is gebruik om die gasvloei deur inlaat en uitlaat sisteme van 'n enjin te analiseer. Die teoretiese afleiding van bestendige gasdinamiese randvoorwaardes asook hul integrasie met die bestendige pypvloei, word verduidelik. 'n Simulasie vloeimodel is ontwikkelom die vloei met behulp van die metode van karakteristieke te analiseer. Hierdie vloeimodel is deel van 'n omvattende enjinsimulasie program, ESA. Dit word gebruik om binnebrandenjins te simuleer en enjinwerkverrigting te voorspel. 'n Nokprofielmodel en 'n termodinamiese ontbrandingsmodel word gebruik om die enjinsimulasie program af te rond. Eksperimentele toetse op 'n gemodifiseerde Nissan Z24/NA20 enjin is gebruik om die simulasie model te evalueer. Die spruitstukke van die Nissan Z24/NA20 is aangepas om een van die silinders te isoleer om so 'n geskikte enkelsilindermodel te skep. Verdere eksperimentele toetse is gedoen op Volkswagen 1.6£8- klep en 1.6£ 20-klep enjins. Werkverrigtingsdata is verkry op twee nuwe inlaatspruitstukke wat met behulp van die ESAsagteware ontwerp is. Werkverrigtingsdata en drukverdelingsdata in die inlaatspruitstuk van die Nissan Z24/NA20is aangeteken om te vergelyk met die resultate van die ESAsagteware. Goeie ooreenstemming is verkry tussen toets- en gemoduleerde data. Die verskille varieer tussen ±5% op enjin werkverrigtingsdata en drukpulsfrekwensie voorspellings, en ± 10%op drukpuls-amplitudes.

This thesis describes an investigation into the spatial species conversion profiles of a Cu-zeolite SCR under engine conditions at low exhaust gas temperatures; this was then compared with a CFD model that models the catalyst via a porous medium measuring 5 x 5 x 91 cells assuming a uniform cross-sectional flow distribution. Species conversion rates were sampled at fixed points in the axial direction. The analysis of the spatial conversion profiles is a more rigorous method in assessing the ability of a mathematical model to predict the experimental data. It can also assist in the optimisation of the catalyst size, minimising packaging requirements and manufacturing costs. The experiments were undertaken on a light-duty diesel engine at a speed of 1500rpm, and at a load of 6bar BMEP; this provided exhaust gas temeraqtures between 200 and 220°C. NO2:NOx ratios were controlled by changing the size and position of the diesel oxidation catalyst, the inlet NH3: NOx ratio was also also varied, ammonia gas was used instead of urea for the purposes of simlicity. The advantage of testing on an actual engine over lab-babed studies is that the conditions such as exhaust gas composition are more realistic. A 1D CFD model was constructed using the ‘porous medium approach’ with kinetics obtained from open literature. Results from the simulations were then compared with the experimental data for the same engine conditions. It was observed that the majority of the NOx conversion took place in the first half of the brick for all NH3: NOx ratios investigated, and that the formation of N2O via NO2 and ammonia had the same influence as the ‘fast’ SCR reaction just after the inlet, which the CFD model failed to predict for the base case analyses. The influence of the inlet ammonia on the model was also noticed to be greater than in the experiments. Simple transient analyses were also undertaken on the short SCR bricks for NO2: NOx ratios of 0.6 and 0.07, and it was observed that the response time to steady-state was noticeably higher in the experiments than in the model. Modifications made to the model, including decreasing the influence of the ‘fast’ SCR reaction, and the addition of an empirical term onto the ammonia adsorption provided a noticeably better agreement for different NH3: NOx injection ratios. The desorption kinetics in the model were also altered by increasing the strength of the bonding of the ammonia onto the adsorption sites. This improved the transient agreement between the model and the experiments, but reduced the steady-state concentrations at the exit of the brick for all NH3:NOx ratios investigated.

Thesis (PhD (Electric and Electronic Engineering))--University of Stellenbosch, 2006. / The focus of this thesis is on the critical evaluation of the torque performance of the optimum designed reluctance DC machine (RDCM). The thesis focuses on multi-phase RDCM drives with normal laminated salient pole rotors allowing for high power and high-speed applications. An RDCM is a normal reluctance synchronous machine (RSM) but with direct control of flux and torque just as in brush DC machines. Flux and torque of the RDCM are controlled directly by the use of special phase current waveforms. Specific attention is given in the thesis for the selection of the best current waveform for the RDCM allowing for smooth rotating airgap MMF and less ripple torque. The absolute optimum designed RDCM can best be obtained by the use of the finite element (FE) method in the design optimisation process. In this thesis a multi-dimensional FE based design optimisation method for the optimum design of the current controlled RDCM is implemented. To compare the torque performance of the RDCM with other RSMs the torque performances of optimum designed 3-phase, 5-phase and 5-phase with the injection of third harmonic current RSMs are performed under the same copper losses and stack volume. The torque performances of RSMs are done with both salient pole rotor and the round rotor with internal flux barriers. The armature reaction effect of 6-phase RDCMs is also investigated in detail by considering three different rotor structures. These rotor structures are the standard salient pole rotor, the salient pole rotor with slitted poles and the salient pole rotor with chamfered poles. It was shown that the RDCM with the salient pole rotor has a severe armature reaction effect, which can be reduced by slitted or chamfered salient pole rotors. A per-phase equivalent circuit model of the 6-phase RDCM is also proposed in this thesis. The torque of the machine is calculated based on the per-phase equivalent model and compared with the torque calculated by the FE Maxwell stress tensor method. There is a good agreement between these calculated torques. This thesis shows that the implemented FE based optimisation method can be applied with success to optimally design current controlled RDCMs. It was found, amongst other things, that the torque performance of the optimum designed 6-phase RDCM is slightly higher than that of the optimum designed 5-phase RSM with the injection of 3rd harmonic currents and with the same copper losses and stack volume. The analytical and FE calculated results are confirmed by measured results on a 35 kW 6-phase RDCM drive.

Thesis (PhD (Electrical and Electronic Engineering))--University of Stellenbosch, 2009.

Thesis (PhD (Mechanical and Mechatronic Engineering))--University of Stellenbosch, 2006. / The research described in this dissertation examined the effects that fuel formulation can have on the regulated exhaust emissions produced by spark ignition engines in a South African context. Typical South African engine technology, and fuels representative of available fuels were investigated. To broaden the scope and provide information on as many fuel parameters as possible, fuel formulations other than typical retail fuels were also investigated. In order to gain insight into the mechanisms taking place, combustion analysis was performed on measured cylinder pressure traces