The 90-KDa heat shock protein (Hsp90) is part of the molecular chaperone family, and as such it is involved in the regulation of protein homeostasis within cells. Specifically, Hsp90 aids in the folding of nascent proteins and re-folding of denatured proteins. It also plays an important role in the prevention of protein aggregation. Hsp90’s functionality is attributed to its several staged, multi-conformational ATPase cycle, in which associated client proteins are bound and released. Hsp90 is known to be associated with a wide array of client proteins, some of which are thought to be involved in multiple oncogenic processes. Indeed Hsp90 is known to be directly involved in perpetuating the stability and function of multiple mutated, chimeric and over-expressed signalling proteins that are known to promote the growth and survival of cancer cells. Hsp90 inhibitors are thus thought to be promising therapeutic agents for cancer treatment. A lack of a 3D structure of human Hsp90 however has restricted Hsp90 inhibitor development in large to in vivo investigations. This study, aims to investigate and calculate hypothetical homology models of the full human Hsp90 protein, and to probe these structural models for novel drug target sites using several in silico techniques. A multi-template homology modelling methodology was developed and in conjunction with protein-protein docking techniques, two functionally important human Hsp90 structural models were calculated; the nucleotide free “v-like” open and nucleotide bound closed conformations. Based on the conservation of ligand binding, virtual screening experiments conducted on both models using 316 natural compounds indigenous to South Africa, revealed three novel putative target sites. Two binding pockets in close association with important Hsp90-Hop interaction residues and a single binding pocket on the dimerization interface in the C-terminal domain. Targeted molecular docking experiments at these sites revealed two compounds (721395-11-5 and 264624-39-7) as putative inhibitors, both showing strong binding affinities for at least one of the three investigated target sites. Furthermore both compounds were found to only violate one Lipinski’s rules, suggesting their potential as candidates for further drug development. The combined work described here provides a putative platform for the development of next generation inhibitors of human Hsp90.

On the basis of clinical studies, some researchers have advocated that the neurohormone and antioxidant melatonin, shown to possess intrinsic anticancer properties, be used as co-therapy in cancer patients being treated with the antineoplastic agent 5-fluorouracil, as increased patient survival times and enhanced quality of life have been observed. The focus of this research was thus to investigate the mechanisms of this seemingly beneficial drug interaction between 5-fluorouracil and melatonin. Metabolism studies were undertaken, in which it was established that there is no hepatic metabolic drug interaction between these agents by cytochrome P450, and that neither agent alters the activity of this enzyme system. Co-therapy with melatonin is thus unlikely to alter plasma levels of 5-fluorouracil by this mechanism. Novel mechanisms by which 5-fluorouracil is toxic were elucidated, such as the induction of lipid peroxidation, due to the formation of reactive oxygen species; decreases in brain serotonin, dopamine and norepinephrine levels, possibly leading to depression; hippocampal shrinkage and morphological alterations and lysis of hippocampal cells, which may underlie cognitive impairment; and a reduction in the nociceptive threshold when administered acutely. All these deleterious effects are attenuated by the co-administration of melatonin, suggesting that the agent exhibits antidepressive and analgesic properties, in addition to its known antioxidative and free radical-scavenging abilities. This suggests that melatonin cotherapy can significantly decrease 5-fluorouracil-induced toxicity, but this may also exert a protective effect on cancer cells and thus compromise the anticancer efficacy of 5-fluorouracil. It was, furthermore, found that stimulation of indoleamine 2,3-dioxygenase activity, mediated by increases in superoxide anion and interferon-γ levels, may underlie resistance to 5-fluorouracil therapy. Melatonin was shown to increase superoxide anion levels in vivo, and this is believed to be by conversion to the metabolite and known oxidant 6- hydroxymelatonin. This highlights that the possible deleterious effects of melatonin metabolites should be studied further. Serum corticosterone levels and cytokine profiles are unaltered by both 5-FU and melatonin, suggesting that these agents may be used by HIV infected individuals without promoting the progression to AIDS. It can thus be concluded that melatonin co-therapy is potentially useful in countering 5-fluorouracil toxicity.

Breast cancer is a leading cause of cancer death in Africa. Hsp90 has been identified as a target for anti-cancer treatments as its inhibition results in the disruption and ubiquitin–proteasome degradation of activated oncoproteins. Currently, there are no US Food and Drug Administration approved Hsp90 inhibitor drugs and existing Hsp90 inhibitors such as geldanamycin and novobiocin are hepatotoxic and display a low affinity for Hsp90, respectively. Therefore, there is a need for the development of Hsp90 inhibitors with improved inhibitory properties. In this study twelve natural compounds bearing a quinone nucleus were screened and characterised for the modulation of Hsp90. The compounds analysed formed three series; the sargaquinoic acid (SQA), naphthoquinone, and pyrroloiminoquinone alkaloid series. Certain compounds exhibited half maximal inhibitory concentrations of between 3.32 μM and 12.4 μM, while others showed no antiproliferative activity at concentrations of up to 500 μM in the MDA-MB-231 breast adenocarcinoma cell line. Immunofluorescence and Western analyses indicated that the modulation of Hsp90 and partner proteins by SQA was more similar to that of novobiocin. Isothermal titration calorimetry analyses suggested that SQA interacted with Hsp90β with a low affinity, and saturation-transfer difference nuclear magnetic resonance confirmed that this interaction with Hsp90β occurred through the methyl moiety bound to 1, 4 benzoquinone of SQA. Pulldown assays indicated SQA disrupted the association between Hsp90 and Hop dose-dependently, more similarly to novobiocin. Immunofluorescence and Western analyses performed on naphthoquinone and pyrroloiminoquinone alkaloid compounds indicated modulation of Hsp90 and Hsp90 partner proteins by the compounds. Naphthoquinone compounds were prioritised for analysis for binding to Hsp90β over the pyrroloiminoquinone alkaloid compounds. Lapachol interacted with Hsp90β with a low affinity however; this interaction was thought to be too weak to disrupt the association of Hsp90 and Hop. The remaining naphthoquinone compounds showed no interaction with Hsp90β, thus allowing the determination of a preliminary structure-activity relationship for these compounds. To the best of our knowledge, this is the first study to describe a systematic subcellular analysis of the effects of geldanamycin and novobiocin in comparison to sargaquinoic acid and compounds of the naphthoquinone and pyrroloquinoline scaffold on Hsp90 and its partner proteins. / Microsoft� Word 2010 / Adobe Acrobat 9.54 Paper Capture Plug-in

Breast cancer, the most common malignancy diagnosed in women, is one of the leading causes of death in women worldwide. In South Africa only 32% of women diagnosed with advanced breast cancer survive more than five years. The search for new chemotherapeutic agents capable of effectively treating breast cancer is therefore essential. Recent evidence supporting the cancer stem cell theory of cancer development for breast cancer challenges the current theories of cancer development and hence treatment. Cancer stem cells are a small subpopulation of tumour cells that possess properties of both cancer cells and stem cells and are believed to be the tumour-initiating population of many cancers. Cancer stem cells are inherently resistant to many chemotherapeutic agents and in this way have been associated with repopulation of tumours after chemotherapy. This phenomenon is proposed as a possible mechanism for cancer relapse after treatment. Cancer stem cells have also been implicated in metastasis, the major cause of mortality in cancer patients. Therefore, any treatment that is capable of targeting and removing breast cancer stem cells may have the theoretical potential to effectively treat breast cancer. However, there are currently no such treatments available for clinical use. We were provided access to a library of novel indigenous small molecules isolated from red and brown algae found off the Eastern Cape of South Africa. The aim of this project was to analyse the anti-cancer and anti-cancer stem cell properties of the compounds in this library and to identify „hit‟ compounds which could form the basis for future development into new anti-cancer drugs. Ten novel compounds of algal origin were tested for cytotoxicity, by determining their ability to inhibit the growth of MCF12A breast epithelial cells and MCF7 breast cancer cells using the colorimetric MTT [(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] cell proliferation assay. All but one of the compounds tested exhibited cytotoxicity towards the MCF7 cancer cell line, with IC50 values (the concentration of the compound that leads to a 50% inhibition in cell growth) of between 3 μM and 90 μM. The chemotherapeutic drug paclitaxel was used as a positive control. Four of the compounds (RUMB-001, RUMB-002, RUMB-007 and RUMB-010/saragaquinoic acid) were significantly more toxic to the MCF7 cancer cell line, than the „normal‟ MCF12A breast cells and were selected as priority compounds for further analyses. In addition, two other compounds were selected as priority compounds, one highly cytotoxic towards both MCF12A and MCF7 cell lines (RUMB-015) and one which was non toxic to either cell line (RUMB-017/018). Preliminary studies into the mechanism of cytotoxicity using Western blot analysis for poly (ADP-ribose) polymerase (PARP) cleavage and Hoechst 33342 immunostaining in MCF-7 cells were largely unsuccessful. The Hoechst 33342 immunostaining assay did provide tentative evidence that selected priority compounds were capable of inducing apoptosis, although these assays will need to be repeated using a less subjective assay to confirm the results. The priority compounds were subsequently investigated for their cytotoxic effect on the cancer stem cell-enriched side population in MCF7 cells. The ability of the priority compounds to selectively target the cancer stem cell containing side population was assessed using two complementary flow cytometry-based techniques – namely the Hoechst 33342-exclusion assay, and fluorescent immunostaining for the expression of the putative cancer stem cell marker, ABCG2+. The ABCG2+ staining assay was a novel technique developed during the course of this study. It remains to be fully validated, but it may provide a new and reliable way to identify and analyse cancer stem cell containing side population cells. The MCF7 cells were treated with the compounds and the proportion of putative cancer stem cells compared with the size of the population in untreated cells was assessed. Three compounds (RUMB-010, RUMB-015 and RUMB-017/018) capable of reducing the proportion of side population cells within the MCF7 cell line were identified. Taking these data together, we identified two potential „hit‟ compounds which should be prioritised for future research. These are compounds RUMB-010/sargaquinoic acid and RUMB-017/018. RUMB-010 is of interest as it was shown to target the putative cancer stem cell population, in addition to the bulk MCF7 tumour line, but was relatively less toxic to the „normal‟ MCF12A cell line. RUMB-017/018 is of interest due to the ability to selectively target the cancer stem cell enriched side population, while having little effect on the normal (MCF12A) or bulk tumour (MCF7) cell lines tested. These compounds will be important as „hit‟ compounds for drug development and as tool compounds to study cancer and cancer stem cell biology.

The effect of the inclusion of sexual behaviour, particularly three sexual orientation classes, on the transmission dynamics of HPV and cervical cancer incidence was investigated. A comprehensive literature review of mathematical models of HPV transmission and the natural history of cervical cancer was concluded. A mathematical model using ordinary differential equations was developed, which incorporated the three sexual orientation classes, and a sexual mixing algorithm for modelling the transmission dynamics. Reproduction numbers, determined through a simplified version of the developed model, indicated that the bisexual population could form a bridge between the heterosexual and homosexual population. The level of interaction is determined by the selection preferences of a bisexual individual to form a partnership with an individual of the same or opposite sex. The model was simulated, with parameters based on a South African population and HPV type 16/18, to investigate the effects of HIV status, sexual orientation and various vaccination strategies on HPV transmission and cervical cancer incidence. The results indicated that HIV status is a significant factor when determining cervical cancer incidence. The results regarding vaccination strategies agreed with results from the literature review with a two sex before sexual debut and catch up program the most effective, noting that with increased vaccination coverage of females the marginal impact on cervical cancer incidence of this approach diminished.

Cancer is a disease that imposes a heavy burden on public health and poses a challenge to science. The World Health Organization estimates that 80 percent of people in developing countries of the world rely on traditional medicine for their primary health needs, and about 85 percent of traditional medicine involves the use of plant extracts. This is particularly true in Africa where a large percentage of the population depends upon medicinal plants for health care. Therefore, detailed screening and evaluation of bioactive substances for chemotherapeutic purposes of African plants are urgently warranted. Furthermore, this will serve to validate the efficacy and safety of African traditional medicine. The current study investigated the in vitro cytotoxic effects of 17 ethanolic extracts of the following 16 plants used in traditional anticancer medicine in Nigeria: Sapium ellipticum leaves, Sapium ellipticum stembark, Combretum paniculatum, Celosia trigyna, Pupalia lappacea, Justica extensa, Hedranthera barteri leaves, Alternanthera sessilis, Ethulia conyzoides leaves, Lannea nigritana stembark, Combretum zenkeri root, Combretum molle leaves, Adenanthera parvoniana, Lannea acida, Cyathula achyranthoides, Drymaria cordata, Cyathula prostrata, against HeLa cancer cells. Five of the most promising extracts (Sapium ellipticum leaves, Combretum paniculatum, Celosia trigyna, Drymaria cordata, Cyathula prostrata) were selected for further screening against HT29 and MCF-7 cancer cells. Of the five, the first two were investigated further based on their activities in the screening phase. The S. ellipticum leaf extract yielded IC50 values of 88.60 ± 0.03 and 93.03 ± 0.03 μg/ml against HeLa and MCF-7, respectively. The toxicity was also evaluated on normal cells and an IC50 of 77.66 μg/ml was obtained for peripheral blood mononuclear cells (PBMCs). The IC50 values for proliferating and confluent Chang liver cells were both >125 μg/ml. These results suggest that the extract may be selective for specific cell types. Bio-assay guided fractionation of the S. ellipticum ethanolic extract yielded two active fractions; chloroform and ethyl acetate. Two compounds isolated from the chloroform extract were screened against the three cancer cell lines and found to be inactive. Three compounds were isolated from the ethyl acetate fraction and revealed IC50 values < 62.5 and < 31 μg/ml against MCF-7. Unfortunately these two compounds soon lost activity before any further work could be done on them and work was continued with the crude extract.

As breast cancer incidence and survival rates increase, there is an urgent need to make appropriate psychosocial support available to all women with breast cancer. In this qualitative study, narrative inquiry was used to examine how women with breast cancer used visual creative expression (art therapy and/or independent art making) to address psychosocial needs that arose for them after their diagnoses. Seventeen women, aged 37-82, participated in this investigation. Data analysis of in-depth interviews with these women focused on narratives they constructed about why they turned to art therapy and/or independent art making and how it helped to be involved in these activities. Particular attention was given to the issue of meaning making. Four storylines emerged from the analysis. "Art and art therapy as a haven" came from narratives about using art making or art therapy for comfort and affirmation. The narratives that comprised "getting a clearer view" were about using visual creative expression to create a clear picture of emotional experience. "Clearing the way emotionally" came from narratives about self-expression and about processing difficult emotions. The narratives that yielded "expanding and enlivening the self were about the women fortifying and energizing themselves through visual creative expression. Two minor themes related to the role of the art therapist and negative experiences with art therapy also emerged. In their narratives, the women portrayed visual creative expression as flexible, compelling, and powerful means of addressing multiple psychosocial needs simultaneously. Above all, the storylines show that the women valued visual creative expression as a way to reduce the feeling of threat to existence, to affirm present existence, and to promote the ongoing existence of both their psyches and their bodies. The results of this study contribute to the field of psycho-oncology by extending understandings of meaning making in relation to breast cancer, supplying detailed explanations from the perspectives of women with breast cancer of how visual creative expression can be helpful, and providing valuable insight into how psychosocial support services based on visual creative expression might meet needs of women with breast cancer that would not be met through other types of services. / Graduate and Postdoctoral Studies / Graduate

The traditional model of leadership in medicine and health care generally centres around a hierarchical structure of power and influence, resting in the hands of a select few administrators, with limited input from employees. A newly developed Cancer treatment centre in the Province of British Columbia, Canada has attempted to institute a unique, team-based system of shared leadership and decision-making. In order to accomplish this task, the Senior Administrator of the centre hired professional group development experts to facilitate the formation of the newly established Leadership Team. A team of nine individuals participated in a group-process based model of team leadership development, consisting of a series of intensive weekend workshops. This study evaluates the impact of those intensive workshops on the members of the Cancer centre Leadership Team. Qualitative case-study methodology, combined with the use of indepth interviews, illuminated eight categories of shared experience among seven of the nine team members, as a result of having participated in the workshop series. / Education, Faculty of / Educational and Counselling Psychology, and Special Education (ECPS), Department of / Graduate

A novel biocompatible and biodegradable polymer, termed poly(Glycerol malate co-dodecanedioate) (PGMD), was prepared by thermal condensation method and used for fabrication of nanoparticles (NPs). PGMD NPs were prepared using the single oil emulsion technique and loaded with an imaging/hyperthermia agent (IR820) and a chemotherapeutic agent (doxorubicin, DOX). The size of the void PGMD NPs, IR820-PGMD NPs and DOX-IR820-PGMD NPs were approximately 90 nm, 110 nm, and 125 nm respectively. An acidic environment (pH=5.0) induced higher DOX and IR820 release compared to pH=7.4. DOX release was also enhanced by exposure to laser, which increased the temperature to 42°C. Cytotoxicity of DOX-IR820-PGMD NPs was comparable in MES-SA but was higher in Dx5 cells compared to free DOX plus IR820 (pIn vivomouse studies showed that NP formulation significantly improved the plasma half-life of IR820 after tail vein injection. Significant lower IR820 content was observed in kidney in DOX-IR820-PGMD NP treatment as compared to free IR820 treatment in our biodistribution studies (p

Bakgrund: Cancerbehandlingar kan medföra konsekvenser hos kvinnor så som skador på äggstocksvävnaden vilket kan leda till medicinsk infertilitet. Fertilitetskonservering är en metod för att bevara könsceller vid risk för infertilitet som bör erbjudas till kvinnor vid ett cancerbesked beroende på cancertyp, cancerbehandling och relationsstatus. Syfte: Litteraturstudiens syfte var att belysa kvinnors upplevelse av information om fertilitet och risk för infertilitet relaterat till sin cancerbehandling. Metod: Litteraturstudien genomfördes och grundades på åtta resultatartiklar, fem kvalitativa och tre kvantitativa. Resultat: Det finns en tydlig röd tråd i samtliga resultatartiklar att kvinnor upplevde att informationen relaterat till hur deras fertilitet kunde komma att påverkas av cancerbehandlingen varit bristande eller helt uteblivit. Vid information var upplevelsen ofta negativ och kvinnorna lämnades ovetandes med funderingar och tankar. Resultatet har delats in i två teman: Bristande information och Ojämlik information. Konklusion: Då vi i dagens samhälle har större chanser att överleva cancer än tidigare bör hälso- och sjukvårdspersonal prioritera individen och fokusera på livet efter cancern. Informationen som ges till patienter bör individanpassas för att ge patienter den bästa förutsättningen till familjebildning efter cancerbehandlingen. / Background: Cancer treatments can have many consequences in women where damage to their ovarian tissue can lead to medical infertility. Fertilitypreservation is a method that should be offered to women undergoing cancer treatment to preserve gametes in women in risk of becoming infertile depending on cancer diagnosis, cancer treatment and relationship status. Aim: The aim of the literature study was to illuminate women's experiences receiving information regarding fertility and risk of infertility during a cancer treatment. Method: The literature study was based on five qualitative and three quantitative studies. Result: There's a distinct red line in women's experiences of lack off or no information at all related to how fertility could be affected by a cancer treatment. When informed the experience was mostly negative and women were left unknowing with reflections and thoughts. Out of the eight studies two main themes emerged: Lack of information and Non equal information. Conclusion: Because of the chances of cancer survival in todays society are higher than before the healthcare community needs to prioritize individuals and focus on life after cancer. Information given should be individualized for patients to enable the best conditions for family planning after a cancer treatment.