Submitted a requirements for the degree of Master of Medicine in the branch of Community Health (Occupational Medicine). It has not been used, either wholly or partially, for any other degree or examination at this or any other university. / Introduction This is the first study in South Africa to look for an association between breast cancer and occupations in black women. Breast cancer is one of the commonest forms of malignancy experienced by women in South Africa and its incidence is increasing (1). Approximately six million women work in South Africa, some of these women are likely to be exposed to carcinogenic agents during their daily working lives. Many occupational carcinogens to the breast have been described and there is some evidence suggesting that many more synthetic chemicals used in different industries may also have carcinogenic properties that have not fully been explored as yet (2). This study plans to identify occupations that place black women at risk of breast cancer in the South African context. Thereafter, it will assess if there is an association between shift work (a known carcinogen to the breast) and breast cancer in black women in South Africa. The findings of the study could be of particular significance to the local context, given that women are entering the workforce in increasing numbers possibly putting more women at risk. Aims The aims of the study are to identify occupations that possibly increase the risk of breast cancer in South Africa, and to specifically assess if there is an association between shift work and breast cancer in black South African women. Objectives To determine whether there are associations between different occupations and breast cancer in black women by calculating the odds ratios (ORs) for breast cancer in different occupations in South Africa. To examine the association between shift work and breast cancer in black women adjusting for variables that may confound the association. Methods The study is an unmatched case-control study using secondary data from the existing Johannesburg Cancer Case Control Study (JCCS) database. The JCCS study is a large ongoing study that recruits black male and female cancer patients with all types of cancers receiving treatment at Charlotte Maxeke Johannesburg Academic Hospital (CMJAH), a tertiary hospital in Johannesburg. All women patients recruited from 1 January 2001 to 31 December 2009 were included in the analysis. This included 1 903 cases and 3 990 controls. An expert group was set up to estimate the likelihood of occupational engagement in shift work for the existing occupational groupings present in the JCCS database. These occupations were classified into: a) high probability of shift work; b) possibility of shift work and c) unlikely to do shift work. ORs were estimated using logistic regression. Those who had never worked were the reference category. Bivariate analysis was then carried out to estimate ORs between individual occupations and breast cancer and later to estimate ORs between the likelihood of involvement in shift work and breast cancer. Multivariate logistic regression followed a forward stepwise approach and all the possible confounders present in the JCCS database were considered. These included age, smoking, drinking alcohol, age at first menarche, parity, age at menopause, use of oral contraception and retroviral status. Results With regards to occupation, the bivariate analyses showed significant ORs for breast cancer in the occupational categories of health, education, social services, retail, hospitality, construction, food, textile and manufacturing, with the highest OR in education (OR 2.33, 95% CI: 1.44 – 3.78) and social service (OR 2.39, 95% CI: 1.24 – 4.58) followed by office workers (OR 2.17, 95% CI: 1.47-3.20) and health workers (OR 2.01, 95% CI: 1.33-3.04). Agriculture (OR 0.55, 95% CI: 0.32 – 0.94) and domestic workers (OR 0.96, 95% CI: 0.75 – 1.22) had ORs under one. Following the adjustments for possible confounders, no statistically significant ORs were found between specific occupations and breast cancer. However, manufacturing had the highest OR (OR 1.44, 95% CI: 0.42- 4.94), followed by office workers (OR 1.44 95% CI: 0.31- 5.94) and health workers (OR 1.31, 95% CI: 0.36-4.76) as compared to the never worked group. In the bivariate analyses there was an association between a possibility of doing shift workers and breast cancer (OR 1.66, 95% CI: 1.41-1.97). Similarly, women who had a high likelihood of participating in shift work had an elevated odds ratio for the disease (OR 1.76, 95% CI: 1.44-2.15). After adjusting for confounders in the multivariate model the ORs for both shift work groups were increased with women who possibly did shift work having an OR of 2.18 (95% CI: 1.34- 3.56) and those who had a high likelihood of carrying out shift work having an OR of 2.13 (95% CI: 1.26- 3.61). Conclusions The bivariate analyses identified elevated ORs for breast cancer in some occupations but in multivariate analyses no statistically significant OR were found. Nevertheless ORs>1 were found for the occupational categories of manufacturing, office and health workers. This study reports a strong association of breast cancer and shift work, which is supported by the literature. However shift work still remains necessary for the functioning of many industries. Many gaps still exist, however, and this study has tried to address one of the neglected areas of occupational risks for breast cancer development.

A dissertation submitted to the Faculty of Science, University of Witwatersrand, Johannesburg, in partial fulfilment of the requirements for the degree of Master of Science. Gauteng, Johannesburg, 2013. / Retinoblastoma Binding Protein 6 (RBBP6) is a multi-domain protein that uses its ring finger domain to interact with p53 and pRb tumour suppressor genes. The mechanism by which RBBP6 uses to degrade p53 is still unknown. Nonetheless, it is well known that RBBP6 promotes cell proliferation in several cancers by negatively regulating p53 via its E3 ubiquitin ligase activity (Ntwasa, 2008). Degradation of p53 by RBBP6 may compromise p53-mediated apoptosis in breast cancer. This study is intended to investigate the potential applications of RNA interference (RNAi) to block RBBP6 expression as well as its subsequent effect on cell growth and apoptosis. To achieve these methodologies, the following techniques were used: RT-PCR, western blotting, xCELLigence system and flow cytometry. Our studies indicate that the knockdown of RBBP6 expression by siRNA modulates p53 gene involved in cell death pathways and apoptosis, showing statistically significant gene expression differences. RBBP6siRNA significantly reduced cell index (CI) compared to the control samples and we observed an inhibition of cellular proliferation in the interval of between 24 and 48 h, as shown in the data obtained by dynamic evaluation using the xCELLigence System. These results were further confirmed by flow cytometry which showed some apoptotic activity. About 20.7% increase in apoptosis was observed in cells co-treated with RBBP6 siRNA and camptothecin when compared to camptothecin-only whereas in siRBBP6 and Staurosporine treated there was only 8.8% increase in apoptosis. These findings suggest that silencing RBBP6 may be a novel strategy to promote staurosporine- and camptothecin-induced apoptosis in breast cancer cells. Keywords: Retinoblastoma Binding Protein 6, staurosporine, camptothecin

MSc (Med), Faculty of Health Sciences, University of the Witwatersrand / The HER2 gene is amplified in 20-30% of breast cancers, a common cancer amongst South African women. HER2 amplification is associated with a poor prognosis and predicts response to treatments such as Herceptin. The gold standard for HER2 testing is Fluorescent in situ Hybridisation (FISH) with dual colour probes for the HER2 gene and chromosome 17 centromere (CEP17) internal control. According to international guidelines, a HER2/CEP17 ratio >2.2 is considered positive. The HER2 FISH test is complicated by the emergence of ambiguous cases with increased CEP17 signals that cannot be accounted for by chromosome 17 polysomy (> 6 copies of CEP17) and that may hide true HER2 gene amplification. The aims of this study were to characterise the HER2 amplicon, in particular the copy number of genes in the vicinity of the HER2 gene, and to design an alternative control probe that could clarify the HER2 gene status in ambiguous cases. In addition, results on 1558 breast cancer specimens sent for routine testing were analysed to determine the trends of HER2 amplification amongst South African women. The rate of HER2 gene amplification was significantly higher (p < 0.05) in African patients (52%) than in Caucasian patients (43%). In Caucasian women, the rate of HER2 amplification in the younger group (68%) was significantly higher (p < 0.05) than in the general Caucasian group (43%), while the same was not seen in the African cohort. Nineteen ambiguous cases with more than 9 copies of CEP17 were further investigated. FISH assays with four different probe kits (PathVysion HER-2: Poseidon Repeat free TOP2A, HER2, CEP17: and Vysis PML-RARA respectively) were performed to determine the copy number of the HER2, TOP2A, RARA genes and CEP17. An in-house dual colour probe kit was designed using the ACTG1 gene as a control for HER2. Of the 19 ambiguous cases, 16 had centromeric amplification, showing that CEP17 is no longer an adequate internal control in FISH HER2 testing. The TOP2A gene was only amplified in HER2 positive cases and the RARA gene was only amplified when the TOP2A gene was also amplified. FISH with ACTG1 as v a control clearly revealed HER2 amplification in ambiguous cases on image analysis and gave HER2/ACTG1 ratios significantly higher than HER2/CEP17 ratios. However, screening of an additional 40 unambiguous cases showed an increased copy number, although limited ( 8), of the ACTG1 gene in four patients; this warrants further testing to assess the value of this gene as a control. Interestingly, a trend was observed for ACTG1 increased copy number in HER2 negative cases, this may point to the presence of a driver gene whose amplification tends to be mutually exclusive from HER2 amplification.

MSc (Med), Community Paediatrics, Faculty of Health Sciences, University of the Witwatersrand / Background: Although breastfeeding is a key child survival strategy, breastfeeding practices in South African health institutions can generally be categorised as being poor. The global Baby-Friendly Hospital Initiative (BFHI), led by UNICEF and the WHO, aims to create a health care environment that promotes breastfeeding as the norm. This study aimed to document practices around breastfeeding support and compliance with the BFHI’s “10 steps to successful breastfeeding” in both baby-friendly accredited and non-accredited facilities. Methods: This was a cross-sectional study, conducted in nine facilities in Gauteng, including tertiary, secondary and districts hospitals, and midwife obstetric units. Convenience sampling was used. Study questionnaires were based on the generic BFHI assessment tool, but were modified to include more detailed investigation of HIV related factors. The questionnaire was verbally administered on-site to 165 mothers of well infants, and 65 nursing staff. Results: Suboptimal practices were identified in both baby-friendly and non-baby-friendly accredited facilities, but more so in the latter. None of the facilities passed all of the 10 BFHI steps. None of the baby-friendly certified institutions achieved a score sufficient to be still deemed baby-friendly. Steps that required advice and support from nursing staff, viz. step 5 (showing mothers how to breastfeed), step 8 (encouraging breastfeeding on demand), and step 10 (breastfeeding support after discharge from the facility), were particularly poorly done in the majority of facilities. Baby-friendly certified institutions were significantly better than non-accredited facilities for steps 2 (training), 4 (initiating breastfeeding within 1 hour), and 7 (rooming-in). Conclusion: Although baby-friendly accredited facilities generally performed better than non-accredited facilities, their performance failed to justify maintenance of their accredited status based on this assessment. However, some positive practices were sustained over time. There is a dire need for greater attention to be directed to the promotion of good breastfeeding practices by health professionals and institutions.

by Yu Xiongwen. / Thesis (M.Sc.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 74-84). / Abstract also in Chinese. / Acknowledgments --- p.i / Abstract --- p.ii / List of Tables --- p.iii / List of Figures --- p.iv / Chapter Chapter 1 --- Introduction --- p.1 / Chapter 1.1 --- Tumor Marker --- p.1 / Chapter 1.1.1 --- General concept of tumor marker --- p.1 / Chapter 1.1.2 --- Application of tumor marker --- p.2 / Chapter 1.1.3 --- Limitation of tumor markers --- p.5 / Chapter 1.2 --- Breast Cancer --- p.6 / Chapter 1.2.1 --- Incidence in Hong Kong Chinese --- p.6 / Chapter 1.2.2 --- Need for early diagnosis and prognosis --- p.8 / Chapter 1.3 --- Markers for Breast Cancer --- p.9 / Chapter 1.3.1 --- Usefulness of tumor marker for breast cancer --- p.9 / Chapter 1.3.2 --- Some tumor marker for breast cancer --- p.12 / Chapter 1.4 --- Selective Markers for Breast Cancer in this Study --- p.16 / Chapter 1.4.1 --- New TPA --- p.16 / Chapter 1.4.2 --- CA 15-3 --- p.19 / Chapter 1.4.3 --- Apolipoprotein(a) --- p.22 / Chapter 1.5 --- Objectives --- p.24 / Chapter Chapter 2 --- Materials and Methods --- p.25 / Chapter 2.1 --- Materials --- p.25 / Chapter 2.1.1 --- Patients and control subjects --- p.25 / Chapter 2.1.2 --- Sampling --- p.25 / Chapter 2.2 --- Methods --- p.26 / Chapter 2.2.1 --- CA 15-3: Cancer Antige 15-3 --- p.26 / Chapter 2.2.2 --- New TP A --- p.27 / Chapter 2.2.3 --- Apolipoprotein(a) --- p.28 / Chapter 2.3 --- Statistical Methods --- p.29 / Chapter Chapter 3 --- Results --- p.32 / Chapter 3.1. --- Precision Studies --- p.32 / Chapter 3.1.1 --- CA 15-3 --- p.32 / Chapter 3.1.2 --- TPA --- p.32 / Chapter 3.1.3 --- Apolipoprotein(a) --- p.32 / Chapter 3.2 --- CA 15-3 --- p.37 / Chapter 3.2.1 --- "CA 15-3 levels in healthy women, patients with benign breast disease and patients with breast cancer" --- p.37 / Chapter 3.2.2 --- "Sensitivity, specificity, and total accuracy of preoperative CA15-3 determination by cutoff value" --- p.42 / Chapter 3.3 --- TPA --- p.45 / Chapter 3.3.1 --- TPA levels in healthy women，patients with benign breast disease and patients with breast cancer --- p.45 / Chapter 3.3.2 --- "Sensitivity, specificity，and total accuracy of preoperative CA 15-3 determination by cutoff value" --- p.50 / Chapter 3.4 --- Apolipoprotein (a) --- p.53 / Chapter 3.4.1 --- Apo(a) levels in healthy women，patients with benign breast disease and patients with breast cancer --- p.53 / Chapter 3.5 --- Combination Test --- p.59 / Chapter 3.6 --- Study in Pairs --- p.64 / Chapter 3.6.1 --- Results of the pairs investigation --- p.64 / Chapter 3.6.2 --- Changes in post-operation compared with the pre- operation levels --- p.64 / Chapter Chapter 4 --- Discussion --- p.69 / Chapter Chapter 5 --- Conclusion --- p.73 / References --- p.74

Matrix Metallopeptidase 1 (MMP-1) expression has repeatedly been correlated to tumorigenesis and metastasis. Yet, MMP-1 regulation in a metastatic context remains largely unknown. Here we confirm differential MMP-1 expression in mammary carcinoma cells with varied metastatic potentials and identify a mechanism differentially regulating MMP-1. We show that MMP-1 expression is regulated by an AP-1 element in its promoter in highly metastatic MDA-MB-231 mammary carcinoma cell derivatives. Fra-1, an AP-1 family transcription factor, differentially binds this element in highly metastatic derivatives compared to low-metastatic cells and is required for MMP1 expression. Fra-1 mRNA levels are unchanged in the cell variants, however its protein levels are higher in the metastatic cells. There was no change in protein degradation rates, while protein synthesis rates of Fra-1 increased. These results suggest that protein translation of Fra-1 is differentially regulated in these cells. Consistent with the importance of Fra-1 for tumor growth, we found that Fra-1 overexpression is sufficient to increase cell motility and anchorage independent growth. These results suggest that Fra-1 regulation is critical for regulation of MMP-1 and metastasis.

No description available.

Breast cancer is a heterogeneous disease, and can be classified according to histological subtypes based on cellular morphology. Invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) are the most common histological subtypes, accounting for approximately 80% and 12% of cases respectively. ILC exhibits a number of distinct clinico-pathological features in comparison with IDC, and is understudied as a breast cancer subtype. ILC tumours are typically oestrogen receptor positive, HER2 negative, and frequently demonstrate early loss of Ecadherin expression, which is a hallmark of the lobular phenotype. ILC is presently treated in a similar manner to IDC, with treatment generally directed against hormone receptors. Upon acquisition of hormone resistance, limited secondary options are available; patients are rarely candidates for agents targeting HER2, and are recognised to be poorly responsive to chemotherapeutics. We therefore need to advance our understanding of lobular tumour biology, in order to identify suitable biomarkers that will guide the development of targeted therapies for ILC patients. As protein expression levels determine cellular phenotype, a protein-based approach has the potential to provide biologically relevant insight into the mechanisms driving ILC. A range of protein analysis platforms, including reverse phase protein array, label-free mass spectrometry and immunohistochemistry, were therefore used to elucidate biological mechanisms active in the ILC subtype. Such experiments led to the identification of activated PI3K-Akt signalling in mouse and human ILC, suggesting that inhibition of this pathway may be an effective treatment strategy in lobular breast cancer. Preliminary evidence of differences in cytoskeletal and extracellular matrix (ECM) proteins was also acquired, providing an interesting basis for future research. A further major strand of this project was the development of in vitro and in vivo tools, to facilitate further interrogation of lobular biology. This included determination of a representative mouse model of ILC, and generation of primary cancer cells and cancer-associated fibroblasts (CAFs) from patient-derived material. Analysis of CAFS showed differential expression of ECM-associated genes, consistent with proteomic analyses. In addition, a tissue micro-array (TMA) comprising primary ILC and IDC tumours, with associated clinical data, was developed. Immunohistochemical staining of the TMA identified a potential role for IGF-1 pathway signalling in ILC, with increased expression of IGF-1 ligand associating with increased tumour size and metastasis in ILC patients. Taken together, the generation and validation of a range of useful tools in the course of this work has provided useful insight into the unique biology of ILC.

No description available.

Breast tumors are not only main incidence of tumor based load of women population, but they are still the most common cause of death in the area of tumor diseases. 1950 women dies on breast tumors every year, which is 37 out of 100 thousand women. Overall are breast tumors cause of deaths in 3.5% women in population. Despite still growing incidence of breast tumors, death rate caused by this type of tumor stagnate on long tearm basis, which clearly shows improvement in treatment success mostly thanks to higher finding in earlier stages. In our country is in 28 years evident shift of incidence into younger age groups, which starts to be obvious in age group of 40-44 years already. Today's prevention possibilities are on high level. It depend's only on patients themselves, if they will obey these rules. At early finding illness of breast carcinoma is prognosis of recovery high. Because of this disease topicality I've decided to process this topic and expand my knowledge.