<p> The properties of the levels of 21Ne and 29Si have been studied via γ-ray angular distribution and linear polarization measurements and γ-γ coincidence studies yielding a number of new spin-parity assignments to the states of both nuclei. Comparison of the results with the Nilsson model for odd nuclei indicate that for 21Ne, good agreement is in general obtained. Nevertheless, a number of interesting discrepancies exist with regard to the negative parity states of 21Ne and explanations have been proposed for some of these. The agreement is also quite good for 29Si with a calculation using a minimum of free parameters, confirming the oblate shape for this nucleus.</p> / Thesis / Doctor of Philosophy (PhD)

Techniques of beta- and gamma-ray scintillation spectroscopy have been applied to a study of the decay radiations from 92.6-min La142. Several new gamma-ray transitions have been discovered and a gamma-gamma coincidence matrix has been established. Nine beta groups have been identified and evidence is presented substantiating the assignment of a first-forbidden unique character to the ground-state beta transition. Angular correlation studies have been performed on some of the gamma cascades. A decay-scheme based on the experimental results is discussed and some spin assignments are made. Some features of the decay modes are interpreted within the context of contemporary ideas about nuclear structure. / Thesis / Doctor of Philosophy (PhD)

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Thesis (MSc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: The aim of this work was to study the 208Pb(p,2p)207Tl quasi-free knockout process. The experimental data were measured at the National Accelerator Centre using incident polarised protons of 200 MeV. The two scattered particles, from the knockout reaction, were detected in coincidence and their energies were determined using a magnetic spectrometer and a solid state detector telescope. Cross section and analysing power energy distributions were extracted from the experimental measurements and these were compared with theoretical values for the Distorted Wave Impulse Approximation. The theoretical cross-section calculations predict the experimental cross-section distribution well for all combinations of distorting potentials and bound states that were investigated, both with regard to shape, as well as absolute magnitude. However the theoretical analysing power distributions did not agree with the experimental quantities. Therefore it is not clear whether the analysing power is a useful tool to extract information on the specifics of the quasi-free reaction mechanism. The spectroscopic factors were found to be consistent with the results obtained in previous studies, thereby inspiring confidence that the problem with the analysing power distribution is not ascribable to a possible deficiency in the experimental techniques exploited in this work. / AFRIKAANSE OPSOMMING: Die doel van die studie was om die kwasi vrye 208pb(p,2p )207TI verstrooingsproses te ondersoek. Die eksperimentele data is ingewin by die Nasionale Versnellingsentrum deur gebruik te maak van 'n 200 MeV gepolariseerde proton bundel. Die twee verstrooide deeltjies is in koïnsidens gemeet. Vir die metings is 'n magnetiese spektrometer en 'n vastetoestand detektorteleskoop gebruik. Die kansvlak- en analiseervermoë-energieverdelings is uit die eksperimentele data verkry en is vergelyk met die berekenings van die Vervormde Golf Impuls Benadering. Die teoretiese kansvlak berekening het die eksperimetele data goed voorspel, vir die verskillende parametrisering van potensiaal en gebonde toestande. Die berekeninge het goed ooreengestem met betrekking tot beide vorm en absolute grootheid. Die berekende analiseervermoë het egter nie goed met die eksperimentele data ooreengestem nie. Dit is dus nie duidelik of die analiseervermoë 'n handige instrument is om inligting oor die betrokke kwasi-vrye reaksie meganisme te bekom nie. Die spektroskopiese faktore was in ooreenstemming met resultate wat in vorige studies verkry is. Dit versterk vertroue dat die probleem met die analiseervermoë nie toegeskryf kan word aan die eksperimentele tegniek wat gebruik is nie.

The eukaryotic interphase nucleus is well organised and the genome positioned non-randomly. Nuclear structure is an important regulator of genome behaviour and function. Genome organisation and nuclear structure are compromised in diseases such as cancer and laminopathies. This study was to find out and to determine if there is any functional relationship between nuclear structure and genome mis-organisation in cancer cells. I have assessed the presence and distribution of specific nuclear structural proteins (A-type, B-type lamins and its receptor LBR, many of their binding proteins such as MAN1, LAP2α, LAP2, and Emerin and other nuclear proteins (PML, Nucleolin, and Ki67) using indirect immunofluorescence. From this study, it is found that the nuclear structure of breast cancer cells is often altered. The most severely affected proteins are the nuclear lamins B1 and B2 and they found as large foci within the nucleoplasm with little LBR expression to localise the lamin B. I also assessed the chromosome positioning (HSA 7, 10, 11, 14 and 17) and gene positioning (AKT1, CCND1, HSP90AA1, EGFR, ERRBB2/HER2 and PTEN) in breast cancer cell lines (T-47D, GI-101, Sk-Br-3 and BT-474) and in normal breast cell lines (MCF-10A) using 2D-FISH technique. I also assessed the position of the genes in nuclei and correlated with gene expression using qRT-PCR. Breast cell lines have treated with a drug named lovastatin and it was found that the cells have restored LBR expression and localisation of lamin B, leading to altered gene positioning and changed expression of breast cancer genes. Since the drug (lovastatin, 12 μM/48 hours) affects the prenylation as a post-translation modification process and lamins B biosythensis, it is found that B-type lamins and its receptor expression and distribution were improved and increased in expression by 2-fold in expression levels in the most affected cells (T-47D, and BT-474) compared to the normal cells (MCF-10A) and these cells also showed abnormal nuclei and dead cells. When analysing the nuclear positioning of the genes (AKT1, HSP90AA1 and ERRBB2/HER2), it is found that AKT1 was positioned periphery in BT-474 and T-47D cells and interiorly in the normal cells (MCF-10A) before treatment whereas the same gene was positioned periphery in T-47D and MCF-10A cells and interiorly in BT-474 after treatment with lovastatin. It is also found that HSP90AA1 was positioned periphery in MCF-10A and T-47D cells and interiorly in BT-474 cells before and after treatment (no change). Moreover, ERRBB2/HER2 gene was positioned periphery in T-47D and BT-474 cells and interiorly in MCF-10A cells before treatment whereas the same gene was positioned periphery in MCF-10A and T-47D cells and interiorly in BT-474 after treatment with the same drug. Regarding LMNB1, LMNB2, and LBR genes, the study focussed only on their expression levels and no work has done on their chromosome positioning as well as gene position before and after treatment. These three genes were over expressed when assessed by measuring the relative and fold changes in expression. Therefore, it is suggestive that 2D-FISH experiment to assess their localisation and their specific chromosome territories is required. The results shown in this thesis demonstrate the importance and roles of nuclear architecture specifically nuclear lamins and the integral nuclear membrane proteins (B-type lamins and LBR) in mediating correct genome organisation and function. The breast normal (immortalised cells) and cancerous cell lines showed different nuclear structures as lamin B affect the position of specific target chromosomes and genes. These results will strength the finding that the nuclear lamina is a significant nuclear structure which associates, organises, and regulates numerous vital nuclear processes and the stability of the genome.

<p>It is well-established that the organization of nuclear components influences gene expression processes, yet little is known about the mechanisms that contribute to the spatial co-ordination of nuclear activities. The salivary gland cells of <i>Chironomus tentans</i> provide a suitable model system for studying gene expression<i> in situ</i>, as they allow for direct visualization of the synthesis, processing and export of a specific protein-coding transcript, the Balbiani ring (BR) pre-mRNA, in a nuclear environment in which chromatin and non-chromatin structures can easily be distinguished. The RNAbinding protein Hrp65 has been identified in this model system as a protein associated with non-chromatin nucleoplasmic fibers, referred to as connecting fibers (CFs). The CFs associate with BR RNP particles in the nucleoplasm, suggesting that Hrp65 is involved in mRNA biogenesis at the post-transcriptional level. However, the function of Hrp65 is not known, nor is the function or the composition of CFs. In the work described in this thesis, we have identified by yeast two-hybrid screening and characterized different proteins that bind to Hrp65. These proteins include a novel hnRNP protein in <i>C. tentans</i> named Hrp59, various isoforms of Hrp65, the splicing- and mRNA export factor HEL/UAP56, and a RING-domain protein of unknown function. Immuno-electron microscopy experiments showed that Hrp59 and HEL are present in CFs, and in larger structures in the nucleoplasm of <i>C. tentans</i> salivary gland cells.</p><p>Hrp59 is a <i>C. tentans</i> homologue of human hnRNP M, and it associates cotranscriptionally with a subset of pre-mRNAs, including its own transcript, in a manner that does not depend quantitatively on the amount of synthesized RNA. Hrp59 accompanies the BR pre-mRNA from the gene to the nuclear envelope, and is released from the BR mRNA at the nuclear pore complex. We have identified the preferred RNA targets of Hrp59 in <i>Drosophila</i> cells, and we have shown that Hrp59 binds preferentially to exonic splicing enhancer sequences.</p><p>Hrp65 self-associates through an evolutionarily conserved domain that can also mediate heterodimerization of Hrp65 homologues. Different isoforms of Hrp65 interact with each other in all possible combinations, and Hrp65 can oligomerize into complexes of at least six molecules. The interaction between different Hrp65 isoforms is crucial for their intracellular localization, and we have discovered a mechanism by which Hrp65-2 is imported into the nucleus through binding to Hrp65-1.</p><p>Hrp65 binds to HEL/UAP56 in <i>C. tentans</i> cells. We have analyzed the distribution of the two proteins on polytene chromosomes and in the nucleoplasm of salivary gland cells, and our results suggest that Hrp65 and HEL become associated during posttranscriptional gene expression events. HEL binds to the BR pre-mRNP cotranscriptionally, and incorporation of HEL into the pre-mRNP does not depend on the location of introns along the BR pre-mRNA. HEL accompanies the BR mRNP to the nuclear pore and is released from the BR mRNP during translocation into the cytoplasm.</p>

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