Research report presented to the department Pharmacy and Pharmacology of the University of the Witwatersrand in partial fulfillment of the requirements of the Masters of Science in Pharmaceutical Affairs, University of the Witwatersrand, 20 February 2013 / The current study was a retrospective observational pilot study designed to assess compliance to bisphosphonates treatment by patients diagnosed with osteoporosis (between 01 January 2010 and 17 August 2011). In this study, the cost of pharmacological agents used in the chronic medical management of osteoporosis was also estimated. The total number of patients whose records were eligible for analysis was 286, with the mean age of 66.7 years. The majority of patient's records (91%) were for females

Thesis (M.S.)--University of Toronto, 1998. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.S.)--University of Toronto, 1998. / Includes bibliographical references.

Thesis (M.S.)--University of Michigan, 2004. / Includes bibliographical references.

Nitrogen-containing bisphosphonates (N-BPs) are the standard treatment in cancerassociated bone disease due to their ability to inhibit osteoclast-mediated resorption. However, beyond their anti-resorpitive activity there is increasing evidence from preclinical studies to demonstrate that N-BPs can also have an anti-tumour effect, although the mechanism is still unclear. In vitro studies suggest that anti-tumour effects may be due to direct effects on tumour cells (by inhibiting protein prenylation) or via indirect effects on other cell types. The studies described in this thesis sought to answer this question by developing novel approaches and molecular tools. N-BP resistant tumour cell lines were created by over-expressing FPP synthase (the molecular target of N- BPs) by lentiviral transduction of B16 melanoma cells. Although these cells were only modestly resistant to N-BP, such an approach might be a novel strategy for determining whether NBPs directly affect tumour cells in vivo. However, further studies carried out with mice bearing 4T1 mammary fat pad tumours demonstrated that macrophages and myeloid derived suppressor cells (MDSC) internalised a fluorescently-labelled N-BP. Since macrophages and MDSC are important for tumour growth and metastasis, these studies indicate that these myeloid cells may actually be responsible for the anti-tumour effects of N-BPs. The functional effects of N- BPs on MDSC and macrophages remain to be determined. However, an in vitro prenylation assay was developed as a novel and sensitive tool to study subtle changes in protein prenylation and should prove vital in future studies to determine conclusively whether N-BPs inhibit protein prenylation in these myeloid cell types in vivo. Finally, proteomic techniques were used to examine the levels of small GTPases in MDA-MB-231 breast cancer cells and showed that Rab7 appears to be elevated in a clone that preferentially metastasises to the skeleton. This highlights the need for further investigation of the role of Rab7 in skeletal metastasis and as a potential new therapeutic target.

A proportion of patients with osteoporosis, Paget’s disease and bone associated cancers show no reduction in disease-indicating markers after treatment with N-BPs, and resistance to bisphosphonates has been documented . We have examined whether polymorphic or isoforms variation in the FPPS gene could influence the ability of N-BPs to inhibit this enzyme and affect the clinical response to treatment. Sequencing revealed a novel (T→C) polymorphism in exon 11, resulting in an amino acid change from Val 364 to A1a (V364A), with an allele frequency of 0.08 for the C allele and 0.92 for the T allele. Kinetic values for isoprenoid substrates GPP and IPP were found to be similar to previously published values. Enzymes did not differ in the IC50 values for inhibition by the N-BPs risedronate, pamidronate, alenodonate, ibandronate and zoledronate. Splice variant analysis revealed the presence of two distinctly spliced and targeted isoforms of FPPS. Overexpression plasmids containing the different isoforms of FPPS were used to elucidate the intracellular localisation of the different isoforms. Furthermore, polyclonal antiserum was produced to use as a specific tool for detection of FPPS. Detection and co-localisation of the peroxisomally targeted isoform of FPPs with the peroxisomes was shown 48h post-transfection, while the mitochondrially targeted isoform of FPPS was undetectable. Overexpression of the cytosolic or peroxisomal isoform of FPPS was unable to induce resistance to N-BPs, even though it has been shown that overexpression of FPPS is in part responsible for the induction of resistance to N-BPs.

Thèse d'exercice : Chirurgie dentaire : Université de Nantes : 2007. / Bibliogr.

Thesis (Ph.D.)--Aberdeen University, 2009. / Title from web page (viewed on Jan. 19, 2010). Includes bibliographical references.

Thesis (doctoral)--University of Oulu, 1993. / Includes bibliographical references.

Thesis (M.Phil.)--Hong Kong University of Science and Technology, 2007. / Includes bibliographical references (leaves 93-105). Also available in electronic version.