Glioma, an infrequent form of cancer, continues to confer poor prognosis despite advances in current therapeutic techniques. This research identified the presence of hsp90a in glioma and investigated its potential as a diagnostic or therapeutic marker. l-Isp90ct is emerging as an encouraging target for cancer therapy attributed to the integral role it plays in the stabilisation and folding of proteins crucial for normal cellular homeostasis and its subsequent involvement in tumorigenesis. Modulation of Hsp90 as a drug target offers the possibility of simultaneously impeding numerous signaling pathways implicated in the development and progression of glioma and has thus far shown evidence of clinical activity in patients with melanoma, breast and prostate cancer. With this in mind, the primary focus of this thesis was to evaluate the association between hsp90a expression in glioma versus the normal brain cell lines and tissues. Results from this study showed that hsp90a was transcribed at a higher level in glioma cell lines and tissues and absent in normal brain cell lines and tissues. This suggests that the expression level of hsp90a may be a distinguishing marker between glioma and normal brain tissue and furthermore, in the subset of glioma cell lines examined, an increase in expression of hsp90cz appeared to correlate with a decrease in malignancy. As modulation of a single target is likely to be insufficient due to the development of resistance, emphasis of research has focused on a combinational attack for optimal therapy. Simultaneous silencing of hsp90a, at transcriptional (siRNA) or post-translational level (RA), and subsequent subjection to a chemotherapeutic drug appeared to be more effective than chemotherapy alone. This increase in chemosensitivity could be attributed to the downregulation of hTERT caused by inhibition of l-Isp90c&. The research has led to two novel findings; 1) The identification of hsp90a in glioma 2) Silencing hsp90a led to approximately 13-fold reduction of TMZ in tissues for the achievement of the same effect with TMZ alone 3) Has led to the following two publications: a). Shervington, A., Cruickshanks, N., Wright, H., Atkinson-Dell, R., Lea, R., Roberts, (3 and Shervington, L. (2006) Glioma: What is the role of c-Myc, Hsp90 and telomerase? MoL celL Biochem. 283, 1-9. b). Shervington, A., Cruickshanks, N., Lea, R., Roberts, (3., Dawson, T and Shervington, L. (2008) Can the lack of Hsp90a protein in brain normal tissue and cell lines, rationalise it as a possible therapeutic target for gtiomas. Cancer Invest. 16, 900-904. As silencing of hsp90a and the subsequent downregulation of /ITERT exhibited a correlation with chemosensitivity, it has further emphasised the need for individual characterisation of tumours to highlight the most effective therapeutic option and improve patient survival rates for glioma.

Following failure to respond to antibiotic therapy, 56 patients (mean age 22yrs; range 15-46yrs) with recorded active Acne vulgaris were treated with a standard course of isotretinoin (1mg/kg body weight for 16 weeks). This study investigated the recovery and analysis of skin surface organisms from several skin sites at the start of treatment, 8 weeks into treatment, at the end of the course and 1 month post treatment. The number of anaerobic isolates - presumptive Propionibacterium acnes and also aerobic isolates recovered by appropriate media from each of three specific sites per patient were compared with age-matched controls of healthy volunteers. Patients and controls exhibited similar total numbers of organisms on the cheek, nares and toe web before treatment. Following treatment with isotretinoin the majority of patients had a significant reduction in bacterial numbers (1-2 orders of magnitude) on the cheek. Numbers in the nares and toe web did not show this reduction. Isotretinoin has no antibacterial activity in vitro or in vivo so the observed reduction would strongly suggest that the effect is mediated through alteration of the skin nutritional micro-environment. The clinic-based patients demonstrated high levels of isolates with antibiotic resistance from the outset; the resistance status of each patients’ skin microbiota was determined before, during and after treatment and the dynamics of the population from these sites was investigated. Previous studies of the nature of skin organisms and of the complex multifactorial mechanisms that lead to the eruption of acne have shown that P. acnes play a significant but as yet poorly explained role in the pathogenesis of the disease although it is traditionally the target organism for anti-acne therapy. The clinical samples were cultured using conventional methods to confirm the presence of P. acnes and staphylococci. Novel modifications of PCR amplification methodology were developed to enhance the detection of specific organisms. New protocols for comprehensive and detailed RAPD-PCR analysis of whole cell PCR were developed and employed to profile recovered isolates from individual patients before and after clinical treatment. RAPD-PCR proved to be a useful tool to survey changing bacterial profiles within the cohort. Isolates from some patients were highly similar to each other at the outset, but displayed increased diversity by the end of therapy. Interestingly, other studies have linked clonal populations of bacteria with high pathogenicity. Some patients appear to have acquired a strain which was not evident at their original sampling, but was present in other patients attending the same clinic. P. acnes persist in the environment and may be found on many inanimate objects, from where they could become a source for transmission. As there is some evidence of acquisition of strains from the environment, this is of considerable significance to the management of dermatology clinics throughout the country.