MMed (Haematology), Faculty of Health Sciences, University of the Witwatersrand / Multiple Myeloma (MM) is a malignancy of plasma cells. The incidence worldwide has been reported to be 3-4/100 000 of the population. The exact aetiology is not known, but several factors have been implicated in the aetio-pathogenesis of the disease. Chromosomal abnormalities are well documented in MM. Their detection is important, as some of the cytogenetic abnormalities such as the 13q deletion are associated with a poor prognosis. Knowledge of the prognostic factors guides the clinician with respect to the appropriate management of the patient. Prior to the use of fluorescence in situ hybridisation (FISH) as a technique for detecting cytogenetic abnormalities in MM, progress was slow in this field because of the difficulty of obtaining analysable metaphases in view of the low proliferative activity of plasma cells. FISH has significantly improved the detection rate over conventional cytogenetics. Objective: The present study set out to determine the proportion of patients with MM who have a detectable chromosome 13q deletion using conventional cytogenetic and FISH analysis. The FISH technique was specifically studied to see if the detection rate of the 13q deletion is improved compared to conventional cytogenetics. Furthermore, the cytogenetic abnormalities detected were correlated with the course of the disease, as well as other parameters of prognostic significance. vi Methods: Bone marrow aspiration specimens were obtained from thirty (30) patients with MM. Both newly and previously diagnosed patients were included. The sample size was however reduced to twenty (20) because of the need to optimise the technique and improve signal detection. Conventional cytogenetic and FISH analysis was performed using the LSI D13S319 DNA probe as the test probe, and the centromeric alpha 11 and 18 as control probes. The analysis was carried out by two observers. Results: In the current study, the detection of chromosomal aberrations was much better with FISH analysis compared to conventional cytogenetics i.e. 25% versus 5%. Of all the patients with chromosomal aberrations, 25% (5/20) had the specific deletion 13q14 (D13S319). Most of our patients (70%) presented with stage III disease. 60% of those were positive for deletion 13q14 (D13S319), i.e 3/5 patients had stage III disease. However, there was no correlation between disease stage and chromosome status, as the majority of the patients presented with advanced stage disease, irrespective of their chromosomal status. Other factors of prognostic significance such as the haemoglobin level, beta-2 microglobulin and creatinine levels were not found to correlate with the presence of the chromosomal aberration but with disease stage. Furthermore, median survival did not correlate with the presence of the chromosomal abnormality. Conclusion: FISH analysis improves the detection rate of chromosomal abnormalities in MM compared to conventional cytogenetics. The prevalence of 13q14 deletion in our patient population is lower than that reported in the

No / Spatial genome organization in the cell nucleus plays a crucial role in the control of genome functions. Our knowledge about spatial genome organization is relying on the advances in gene imaging technologies and the biochemical approaches based on the spatial dependent ligation of the genomic regions. Fluorescent in situ hybridization using specific fluorescent DNA and RNA probes in cells and tissues with the spatially preserved nuclear and genome architecture (3D-FISH) provides a powerful tool for the further advancement of our knowledge about genome structure and functions. Here we describe the 3D-FISH protocols allowing for such an analysis in mammalian tissue in situ including in the skin. These protocols include DNA probe amplification and labeling; tissue fixation; preservation and preparation for hybridization; hybridization of the DNA probes with genomic DNA in the tissue; and post-hybridization tissue sample processing.

O consumo do pescado é importante para a saúde humana pelo fato desse alimento ser fonte de proteína de alta qualidade, vitaminas, nutrientes essenciais e dois ácidos poli-insaturados ômega-3. Entretanto, o pescado também pode apresentar risco à saúde humana se estiver contaminado por substâncias tóxicas. Visando avaliar o teor de elementos essenciais e tóxicos, foram utilizado as técnicas de Análise por Ativação Neutrônica e Espectrometria de Absorção Atômica na determinação dos elementos As, Br, Cd, Cl, Co, Cr, Cs, Fe, Hg, K, Mg, Mn, Na, Pb, Rb, Sb, Se e Zn nos tecidos comestíveis (filés) de seis pescados dentre os mais consumidos na cidade de São Paulo: corvina - Micropogonias furnieri (Desmarest, 1823); pescada - Cynoscion leiarchus (Cuvier, 1830); robalo - Centropomus undecimalis (Bloch, 1792); sardinha - Sardinella brasiliensis (Steindachner, 1879); anchova - Pomatomus saltatrix (Linnaeus, 1766) e tainha - Mugil liza (Valenciennes, 1836). Para validar os métodos analíticos utilizados em relação à exatidão, foram utilizados o índice z e o valor de HorRat em materiais de referência certificados analisados sob as mesmas condições das amostras. Os elementos Br, K e Na foram determinados por duas modalidades de irradiação e o teste-t de Student foi utilizado para verificar se havia ou não diferença significativa entre as técnicas. Na determinação dos elementos essenciais e potencialmente tóxicos, foi possível a construção de banco de dados para os elementos Br, Cl, Co, Cs, Fe, K, Mg, Mn, Na, Rb, Se e Zn para potenciais futuras discussões. Para avaliar se havia ou não risco à segurança alimentar, foram comparado os teores de As, Cd, Hg e Pb com a legislação vigente no Brasil e com valores internacionais. Em relação à segurança alimentar, para os contaminantes Cd e Pb foram obtidos teores abaixo de todos os limites técnicos usados neste trabalho. Para As, a maior parte deve estar na forma orgânica, portanto, a determinação de Hg se mostrou determinante. De forma geral, não haveria perigo no consumo da sardinha, pescada, tainha, corvina e robalo, enquanto que para a anchova foi inconclusivo. / Fish consumption is important for human health because this food is a source of high-quality protein, vitamins, essential nutrients and two kinds of omega-3 polyunsaturated acids. However, fish consumption may present risk to human health if contaminated by toxic substances. With the aim of evaluating the content of essential and toxic elements, the techniques of Instrumental Neutron Activation Analysis and Atomic Absorption Spectrometry were used to determine As, Br, Cd, Cl, Co, Cr, Cs, Fe, Hg, K, Mg, Mn, Na, Pb, Rb, Sb, Se and Zn in the edible tissues (fillet) of six of the most consumed fishes at São Paulo city: whitemouth croaker - Micropogonias furnieri (Desmarest, 1823), smooth weakfish - Cynoscion leiarchus (Cuvier, 1830), common snook - Centropomus undecimalis (Bloch, 1792), Brazilian sardine - Sardinella brasiliensis (Steindachner, 1879), bluefish - Pomatomus saltatrix (Linnaeus, 1766) and lebranche mullet - Mugil liza (Valenciennes, 1836). To validate the analytical methods for accuracy, z-score and HorRat values were applied to certified reference materials analyzed under the same conditions of samples. The elements Br, K and Na were determined by two irradiation modalities, and the Student\'s t-test was used to verify whether there were significant differences between the modalities. In the analysis of essential and potentially toxic elements, it was possible to create a database for Br, Cl, Co, Cs, Fe, K, Mg, Mn, Na, Rb, Se and Zn for potential future discussions. In order to assess food safety risks, the levels of As, Cd, Hg and Pb were compared with the Brazilians law and international values. Regarding food safety, Cd and Pb contents were bellow all technical limits used in this study. As for As, most part of this element may be in organic form, therefore the Hg analysis was determinant. In general, there would be no risk in the consumption of Brazilian sardine, smooth weakfish, lebranche mullet, whitemouth croaker and common snook, while for bluefish it was inconclusive.

This thesis deals with extraction of data from cell images and their visualisation. Cell images are processed by FISH method. It discusses theory of diagnosis evaluation automation and cell features visualization. That concerns image processing, cell nuclei segmentation, feature extraction and data visualization.