歐樹軍. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 238-253) / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in Chinese and English. / Ou Shujun.

Le complexe de liaison de la coiffe des ARN (CBC) joue un rôle essentiel dans leur maturation et déclenche une variété de réactions biochimiques, via son interaction avec différents partenaires. Deux complexes, CBC-ARS2-PHAX (CBCAP), et CBC-ARS2-ZC3H18-NEXT (CBCN), ont récemment été montré comme important pour cibler les ARN vers l'export (CBCAP) ou la dégradation (CBCN). Cependant, les mécanismes par lesquels la sélection se fait pour l'une voie ou l'autre reste mystérieuse. Ainsi, une question majeure qui reste à résoudre est de savoir quand et comment ces complexes sont recrutés sur les ARN. Dans ce travail, j'ai utilisé la procédure du iCLIP (Cross-Linking and Immuno-Precipitation), afin d'identifier les cibles de ces complexes sur l'ensemble du transcriptome humain. J'ai réalisé un iCLIP sur cinq composants de CBCAP et CBCN, et j'ai comparé les résultats à ceux obtenus avec RBM7, un composant de NEXT précédemment étudié par iCLIP. Mes résultats indiquent que: (i) CBP20, ARS2, PHAX et ZC3H18 se lient près de la coiffe des ARN, tandis que RBM7 et MTR4 se lient partout; (ii) CBP20, ARS2, PHAX et ZC3H18 s'associent à un large ensemble d'ARN transcrits par l'ARN polymérase II et montrent une faible sélectivité; (iii) la liaison de ces protéines varie avec l'état de maturation des ARN, avec le CBC enrichi sur les ARN matures, tout comme ARS2/PHAX/ZC3H18 et MTR4 (bien que dans une moindre mesure), tandis que RBM7 est préférentiellement lié sur les pre-mRNAs non épissés; (iv) une liaison différentielle de RBM7 et MTR4 sur les ARN, avec RBM7 enrichi sur les introns et les PROMPTs, et MTR4 plus présent sur les ARN mature. Bien que des expériences additionnelles soient requises, nous proposons que le CBCAP et le CBCN se lient à un même ensemble d'ARN, ce qui indique à la fois une compétition entre ZC3H18 et PHAX pour la liaison à ces ARN, et l'absence de voies de routage bien déterminées qui ciblerait les ARN vers l'une ou l'autre de ces protéines. Le devenir des ARN pourrait ainsi être déterminé par d'autres caractéristiques des ARN, ou encore par des protéines additionnelles. Ces facteurs pourraient s'allier aux protéines liées à la coiffe afin de favoriser la formation du CBCAP ou du CBCN. Dans le but d’identifier des facteurs additionnels, j'ai réalisé un screen d'interaction par spectrométrie de masse après purification de ARS2 ou CBP80. Ceci a été fait dans des conditions natives ou après un cross-link des complexes à la formaldéhyde, afin de stabiliser les interactions transitoires. Ceci a permis d'identifier de nouveaux partenaires de ARS2 et de CBP80, dont la majorité sont impliqués dans l'épissage des ARN. Des expériences additionnelles seront nécessaires pour valider ces interactions. / The cap-binding complex (CBC) plays a pivotal role in post-transcriptional processing events and orchestrates a variety of metabolic pathways, through association with different interaction partners. Two CBC sub-complexes, the CBC-ARS2-PHAX (CBCAP) and the CBC-nuclear exosome targeting (NEXT) complex (CBCN), were recently shown to target capped RNA either toward export or degradation, but the mechanisms by which they can discriminate between different RNA families and route them toward different metabolic pathways still remain unclear. A major question to be answered is how and when the different CBC subcomplexes are recruited to the RNP. Here, we used an individual nucleotide-resolution UV cross-linking and immunoprecipitation (iCLIP) approach to identify the transcriptome-wide targets for 5 different components of the CBCAP and CBCN complexes, and compared results to the previously analysed NEXT-component RBM7. We report that: (i) CBP20, ARS2, PHAX and ZC3H18 bind close to the cap, while RBM7 and MTR4 bind throughout the mRNA body; (ii) CBP20, ARS2, PHAX and ZC3H18 associate with a broad set of RNA polymerase II (PolII)-derived RNAs and have only mild species preferences; (iii) binding varies with the RNA maturation stage, with the CBC being highly enriched on mature mRNA, ARS2/PHAX/ZC3H18/MTR4 less so, and RMB7 preferentially bound to pre-mRNAs; (iv) MTR4 and RBM7 show different specificities, with RBM7 being highly enriched on introns and promoter upstream transcripts (PROMPTs), while MTR4 is additionally present on mature RNAs. Although more experimental work is needed to fully support our model, we propose that CBCAP and CBCN bind overlapping sets of RNAs, indicating a competition between the proteins ZC3H18 and PHAX, and the lack of a strict RNA sorting mechanism. RNA fate may therefore be determined by additional RNA features and/or by other RNA-binding proteins, which may synergize with the cap and drive the formation of one specific CBC subcomplex instead of another. In an attempt to identify yet unknown factors that may interact with cap-bound CBCAP and CBCN, we performed a protein interaction screen leveraging affinity capture-mass spectrometry (ACMS), using ARS2 and CBP80 as bait proteins. As a complementary approach, we also employed a formaldehyde-based chemical cross-linking strategy, aimed at stabilizing weak/transient interactions. Although we failed to detect any transient interactions involving the CBC, we identified several potential CBC80 and ARS2 interactors, the majority of which are involved in pre-mRNA splicing. Additional quantitative experiments are required to validate our ACMS results and confirm the existence of such protein interactions in vivo.

When the nonconformities are independent, a multivariate control chart for nonconformities called a demerit control chart using a distribution approximation technique called an Edgeworth Expansion, is proposed. For a demerit control chart, an exact control limit can be obtained in special cases, but not in general. A proposed demerit control chart uses an Edgeworth Expansion to approximate the distribution of the demerit statistic and to compute the demerit control limits. A simulation study shows that the proposed method yields reasonably accurate results in determining the distribution of the demerit statistic and hence the control limits, even for small sample sizes. The simulation also shows that the performances of the demerit control chart constructed using the proposed method is very close to the advertised for all sample sizes. Since the demerit control chart statistic is a weighted sum of the nonconformities, naturally the performance of the demerit control chart will depend on the weights assigned to the nonconformities. The method of how to select weights that give the best performance for the demerit control chart has not yet been addressed in the literature. A methodology is proposed to select the weights for a one-sided demerit control chart with and upper control limit using an asymptotic technique. The asymptotic technique does not restrict the nature of the types and classification scheme for the nonconformities and provides an optimal and explicit solution for the weights. In the case presented so far, we assumed that the nonconformities are independent. When the nonconformities are correlated, a multivariate Poisson lognormal probability distribution is used to model the nonconformities. This distribution is able to model both positive and negative correlations among the nonconformities. A different type of multivariate control chart for correlated nonconformities is proposed. The proposed control chart can be applied to nonconformities that have any multivariate distributions whether they be discrete or continuous or something that has characteristics of both, e.g., non-Poisson correlated random variables. The proposed method evaluates the deviation of the observed sample means from pre-defined targets in terms of the density function value of the sample means. The distribution of the control chart test statistic is derived using an approximation technique called a multivariate Edgeworth expansion. For small sample sizes, results show that the proposed control chart is robust to inaccuracies in assumptions about the distribution of the correlated nonconformities. / Graduation date: 2004

published_or_final_version / Medical Sciences / Master / Master of Medical Sciences

M. Tech. Food Technology. / Aims to develop a standardised procedure for the production of soymilk that could be used to evaluate the food technological potential of the 30 different locally grown soybean cultivars. Such a procedure should have standardised soaking and cooking times and temperatures that could ensure the inactivation of lipoxygenase enzymes and other anti-nutritional factors that negatively affect the flavour or nutritional quality of the soymilk. A filtration technique would be needed to quantitatively and reproducibly separate the soymilk and okara fractions. The soymilk produced should have sensory properties that conform to the requirements of a soymilk that is acceptable to the consumer.

M. Tech. Industrial Engineering / The Reconfigurable Vibrating Screen is a newly designed beneficiation machine to be used for mineral particle separation in mines and quarries with adjustable screen capacity and structure in order to respond to changes in production and quantity demand required by the customers. In order to increase the durability, availability, reliability and maintainability of a reconfigurable vibrating screen, an effective method is required to functionally appraise and improve its working condition. The aim of this study is to develop a tool for functionally assessing reconfigurable vibrating screens and existing vibrating screens.

Dissertation submitted in partial compliance with the requirements of the Masters Degree in Technology: Chemistry, in the Faculty of Applied Sciences at the Durban University of Technology, 2008. / In this work, a holistic approach to validate analytical methods was assessed by virtue of Monte Carlo simulations. This approach involves a statement of the methodsâ s scope (i.e. analytes, matrices and concentration levels) and requisites (internal or external); selection of the methodâ s (fit-for-purpose) features; pre-validation and validation of the intermediate accuracy and its assessment by means of Monte Carlo simulations. Validation of the other methodâ s features and a validity statement in terms of a â fit-for-purposeâ decision making, harmonized validation-control-uncertainty statistics and short-term routine work with the aim of proposing virtually â ready-to-useâ methods. The protocol could be transferred to other methods. The main aim is to harmonize the work to be done by research teams and routine laboratories assuming that different aims, strategies and practical viewpoints exist. As a result, the recommended protocol should be seen as a starting point. It is necessary to propose definitive (harmonized) protocols that must be established by international normalisation/accreditation entities. The Quality Assurance (Method verification and Internal Quality Control, IQC) limits, as well as sample uncertainty were estimated consistently with the validated accuracy statistics i.e. E U (E) and RSDi + U (RSDi). Two case studies were used to assess Monte Carlo simulation as a tool for method validation in analytical laboratories, the first involves an indirect polarographic method for determining nitrate in waste water and the second involves a direct determination of heavy metals in sea water by differential pulse anodic stripping voltammetry, as an example of the application of the protocol. In this sense the uncertainty obtained could be used for decision making purposes as it is very tempting to use uncertainty as a commercial argument and in this work it has been shown that the smaller the uncertainty, the better the measurement of the instrument or the laboratoryâ s reputation.

Thesis (M.Tech.: Quality)-Dept. of Operations & Quality Management, Durban Institute of Technology, 2005 xiii, 68 leaves / The increasing awareness and demand of consumers for safe and high quality food have lead many companies to undertake a comprehensive evaluation and reorganisation of their food control systems in order to improve efficiency, rationalisation of human resources and to harmonise approaches. This evaluation in food control systems has resulted towards the necessity to shift from the traditional approach that relied heavily on end-product sampling and inspection and to move towards the implementation of a preventative safety and quality approach, based on risk analysis and on the principles of the hazard analysis critical control point (HACCP) system. Yoghurt is the most popular fermented milk world-wide; the estimated annual consumption in South Africa amounts to nearly 67 million litres. The aim of this study was to implement a HACCP program in a commercial yoghurt factory and then to evaluate the program during certain critical stages of the manufacturing process.

This dissertation reports on the research work of a new business improvement methodology called the design for Six Sigma (DFSS) conducted within a global resources company with specific reference to Manganese Metal Company (MMC), a subsidiary of BHP Billiton (BHPB). The aim of this research was by means of a case study, through action research, to investigate, analyse and evaluate the "Define, Measure, Analyse, Design and Verify" (DMADV) model proposed by Picard (2004) with specific reference to a new product design. The study was concerned with identifying the BHPB strategic business reasons, effects and analysing the financial impact of implementing a DFSS project pertaining to a new product design at MMC. The literature review highlighted that DFSS enhances revenue growth, quality and reliability for a new product. The key findings were that DFSS does not exist within MMC and BHPB, the DFSS methodology could improve and enhance the revenue for a new product design at MMC and none of the BHPB customer sector groups are reporting any operating excellence (OE) annualised cost improvement benefits for DFSS projects. The main recommendation emerging from this research is that an integrated DFSS process will enable BHPB to identify critical leverage points for improving the overall financial performance in achieving the FY09 OE objective of $1 billion in annualised cost improvement benefits thus achieving the world class benchmark standard set by General Electric Corporation.

The increase in trade between nations and relative decrease in international trade barriers has resulted in wider alternatives for customers all over the world. The resulting increase in global competition has caused organisations to use customer focused and internal strategies to remain competitive. One strategy that applies to both external and internal aspects of the organisation is the implementation and use of ISO 9000 quality systems. ISO 9000 is a set of quality guidelines to which an organisation must modify its own quality systems , in order to achieve ISO 9000 accreditation. ISO 9000 is a marketing tool, but there are additional benefits to using an ISO 9000 quality system. There is an opinion that ISO 9000 is not as beneficial as it should be. This exploratory survey of engineering manufacturing organisations in Durban, South Africa, is to prove that commitment and correct implementation of ISO 9000 will result in expected benefits to an organisation. Additional factors such as size of organisation, duration of ISO 9000 accreditation, degree of internationalisation, method of operation and perceived problems are also related to expected benefits of ISO 9000 accreditation. The results of the survey prove that there is a positive correlation between attitudes of commitment, implementation and degree of internationalisation with expected benefits from using ISO 9000 quality systems. It is further recommended that management commitment and training of all employees is required for ISO 9000 to be successful in an organisation. / Thesis (MBA)-University of Natal, Durban, 2001.