M.Tech.

Philosophiae Doctor - PhD / The classical approach to gene discovery has been to culture micro-organisms demonstrating a specific enzyme activity and then to recover the gene of interest through shotgun cloning. The realization that these standard microbiological methods provide limited access to the true microbial biodiversity and therefore the available microbial genetic diversity (collectively termed the Metagenome) has resulted in the development of environmental nucleic acid extraction technologies designed to access this wealth of genetic information, thereby avoiding the limitations of culture dependent genetic exploitation. In this work several gene discovery technologies was employed in an attempt to recover novel bacterial laccase genes (EC 1.10.3.2), a group of enzymes in which considerable biotechnological interest has been expressed. Metagenomic DNA extracted from two organic rich environmental samples was used as the source material for the construction of two genomic DNA libraries. The small insert plasmid based library derived from compost DNA consisted of approximately 106 clones at an average insert size of 2.7Kbp, equivalent to 2.6 Gbp of cloned environmental DNA. A Fosmid based large insert library derived from grape waste DNA consisted of approximately 44000 cfu at an average insert size of 25Kbp (1.1 Gbp cloned DNA). Both libraries were screened for laccase activity but failed to produce novel laccase genes. As an alternative approach, a multicopper oxidase specific PCR detection assay was developed using a laccase positive Streptomyces strain as a model organism. The newly designed primers were used to detect the presence of bacterial multicopper oxidases in environmental samples. This resulted in the identification of nine novel gene fragments showing identity ranging from 37 to 94% to published putative bacterial multicopper oxidase gene sequences. Three clones pMCO6, pMCO8 and pMCO9 were significantly smaller than those typically reported for bacterial laccases and were assigned to a recently described clade of Streptomyces bacterial multicopper oxidases. Two PCR based techniques were employed to attempt the recovery of flanking regions for two of these genes (pMCO7 and pMCO8). The use of TAIL-PCR resulted in the recovery of 90% of the pMCO7 ORF. As an alternative approach the Vectorette™ system was employed to recover the 3’ downstream region of pMCO8. The complexity of the DNA sample proved to be a considerable technical challenge for the implementation of both these techniques. The feasibility of both these approaches were however demonstrated in principle. Finally, in an attempt to expedite the recovery of fulllength copies of these genes a subtractive hybridization magnetic bead capture technique was adapted and employed to recover a full – length putative multicopper oxidase gene from a Streptomyces strain in a proof of concept experiment. The StrepA06pMCO gene fragment was used as a ‘driver’ against fragmented Streptomyces genomic DNA (‘tester’) and resulted in the recovery of a 1215 bp open reading frame. Unexpectedly, this ORF showed only 80% identity to the StrepA06pMCO gene sequence at nucleotide level, and 48% amino acid identity to a putative mco gene derived from a Norcardioides sp JS614. / South Africa

Elementary teachers' spontaneous (unaided) and cued identifications of key concepts in narrative prose were examined. Measures of the influences of exposure to research and attitudes toward the importance of prior knowledge on their cued identifications were investigated. Data were analyzed to determine the degree to which elementary teachers identified cued key concepts and primary teachers' identifications were compared to those of intermediate teachers. Separate and combined measures of teachers' exposure to reading research and attitudes were compared to their cued key concept identifications. A post hoc exploratory content analysis of the spontaneous key concept identifications was undertaken to discover possible patterns or phenomena in the data. Results of the analyses of cued concept identifications indicated: a) teachers were unable to successfully identify key concepts in narrative prose; b) there were no significant differences between primary and intermediate teachers' identifications; and c) exposure to reading research and attitudes towards the importance and use of prior knowledge and concept development influenced teachers' ability to identify key concepts. An examination of spontaneous key concept identifications showed that: a) there was a lack of teacher consensus as to definition of a key concept; and b) teachers were unable to identify passage-relevant key concepts when left to their own resources. / Education, Faculty of / Language and Literacy Education (LLED), Department of / Graduate

M.Tech. (Chiropractic) / The most accepted method of identifying the second sacral spinous process is by using an accepted landmark known as the Posterior Superior Iliac Spines (PSIS). The PSIS lies about 4cm lateral to the second spinous tubercle of the sacrum (Williams and Warwick, 1980). However, in a search of published work, no evidence could be found of any study which established these generally accepted statements (Stonelake, 1983). This was an inter-examiner reliability study aimed to determine whether palpation of the PSIS was a reliable and accurate method of determining the location of the second sacral spinous process (S2) in participants experiencing lumbosacral joint dysfunction. Thirty participants were marked by three examiners in the prone position using the PSIS to locate S2. An x-ray was taken of each participant in the prone position, after being marked by all three examiners. Measurements were then made on the x-rays to assess the location of the markers on the sacral anatomy. The reliability. of palpation of anatomical landmarks needs to be validated further in future studies, as motion palpation and the palpatory assessment of effectiveness of treatment, applied to correct motion restrictions, are predicted- upon accurate localisation of anatomical landmarks (O'Haire and Gibbons, 2000).

No description available.

La mesure des moments magnétiques nucléaires est d'une grande importance pour bien comprendre la structure nucléaire. Le moment magnétique est sensible à la nature de la particule indépendante. Le développement des faisceaux radioactifs permet aujourd'hui d'étudier les spins nucléaires et les moments de noyaux exotiques éloignés de la ligne de stabilité. Cependant, la mesure des moments magnétiques des noyaux radioactifs nécessite le développement des nouvelles méthodes fiables. Le développement réussi de telles méthodes ouvrirait la possibilité de découvrir de nouveaux phénomènes de structure nucléaire. L'étude présentée dans cette thèse est formée de deux expériences. La première expérience a été réalisée à ALTO à Orsay, en France. L'une des principales exigences pour mesurer un moment magnétique nucléaire est de produire un ensemble avec des spins orientés. Ce dernier peut être produit par un mécanisme de réaction approprié et une interaction de spin nucléaire avec le milieu environnant. Le degré d'orientation dépend du processus de formation et du mécanisme de réaction. Le but de cette première expérience était d’étudier le niveau d’orientation du spin nucléaire dans un mécanisme de réaction de fusion incomplète. Le résultat de l'expérience démontre la possibilité d'obtenir un alignement du spin dans une réaction de fusion incomplète d'un ordre de 20%. Ce mécanisme de réaction, avec une telle quantité d’alignement de spin, a le potentiel d’étudier la région riche en neutrons avec des faisceaux radioactifs. La deuxième expérience, et l'essentiel de la thèse, a été réalisée à HIE-ISOLDE au CERN. Cette expérience visait à obtenir des informations de haute précision sur le facteur g d’un état de courte durée. Une nouvelle méthode TDRIV (Time-Differential Recoil In Vacuum) a été appliquée pour la première fois avec des faisceaux radioactifs post-accélérés. La mesure du facteur g était réalisée pour le premier état excité dans un noyau de ²⁸Mg (Eₓ = 1474 (1) keV, T₁/₂ = 1,2 (1) ps). / Knowledge of the nuclear magnetic moments is of great importance to get a clear understanding of nuclear structure. The magnetic moment is sensitive to the single-particle nature of the valence nucleons. The development of radioactive beam facilities allows nowadays studying nuclear spins and moments of exotic nuclei which are far from the stability line. However, the measurement of magnetic moments of exotic nuclei produced as radioactive beams requires the development of reliable methods. Successful development of such methods would open up the possibility to discover new nuclear structure phenomena. The study outlined in this thesis is formed by two experiments. The first experiment was performed at ALTO facility in Orsay, France. One of the main requirements in order to measure a nuclear magnetic moment is to produce a spin-oriented ensemble. The latter can be produced by suitable reaction mechanism and nuclear spin interaction with the surrounding environment. The degree of the orientation depends on the formation process and reaction mechanism. The aim of this first experiment was investigating the level of nuclear spin orientation in incomplete fusion reaction mechanism. Two reaction channels were studied, the isomeric states in ⁶⁵mNi (I = 9/2⁺, Eₓ = 1017 keV, T₁/₂ = 26 ns), and ⁶⁶mCu (I = 6⁻, Eₓ = 563 keV, T₁/₂= 600 ns) with Time-Dependent Perturbed Angular Distribution (TDPAD) method. The result of the experiment demonstrates the possibility of obtaining spin alignment in incomplete fusion reaction of an order of 20%. This reaction mechanism, with such an important amount of spin alignment has potential near radioactive beam facilities to study the neutron-rich region with inverse kinematics reactions. The second experiment, and the main part of the thesis was performed at HIE-ISOLDE at CERN. This experiment aimed to obtain high precision g-factor information on a short-lived picosecond state. A new Time Differential Recoil-In-Vacuum (TDRIV) method was applied for the first time using post-accelerated radioactive beams. The g-factor measurement was performed for the first-excited state in ²⁸Mg nucleus (Eₓ = 1474(1) keV, T₁/₂ = 1.2(1) ps). Since the lifetime of the state is of the order of picoseconds, its g-factor can be measured only via the spin precession of the nucleus in an extremely strong magnetic field (kT). Such fields can only be produced at the nucleus by hyperfine interactions. In order to obtain a high precision on a g-factor measurement, a TDRIV calibration experiment was performed with a stable ²²Ne beam. This run allowed testing the system under the same conditions as with radioactive²⁸Mg beam. In addition, using the known g-factor of the first-excited state in ²²Ne allows to determine the absolute target-to-degrader distance so that to decrease the uncertainty and obtain a high precision g-factor measurement. The obtained calibration parameters from the ²²Ne data will be used in the determination of g-factor of ²⁸Mg.

A thesis submitted in partial fulfillment of the requirements for the degree of Master of Fine Arts in Art: Painting.

No description available.

No description available.

No description available.