The repair of bone defects and fracture non-unions remain a challenge for clinicians. Current materials have limitations regarding quality, availability and there are long-term complications. Alternative graft materials, often in combination with growth factors have been investigated, but, poor retention of growth factors has lead to the clinical use of high doses and/or frequent injections, putting the patient at risk of adverse effects. The aim was to develop and evaluate a controlled bone morphogenetic protein-2 (BMP-2) delivery system based on poly(lactide-co-glycolide) (PLGA) that would improve current therapies by controlling the dose and localisation of BMP-2 at the treatment site as well as providing a biodegradable scaffold to support the newly formed tissue. Reproducible procedures to manufacture spherical PLGA microparticles of defined sizes within a 1-100 µm range were developed. Lysozyme was used to model rhBMP-2 and it was successfully encapsulated with human serum albumin into PLGA microparticles with high entrapment efficiency and retained its activity upon subsequent release. Release rate was manipulated by the use of a PLGA-PEG-PLGA triblock copolymer and resulted in a number of distinctly different but reproducible cumulative release curves which were applicable to the delivery of different growth factors to mimic in-vivo kinetics. A sustained release profile over one month was chosen for rhBMP-2 delivery and it was shown to remain active upon release demonstrated by up-regulation of alkaline phosphate expression in murine C2C12 myoblast cells and human mesenchymal stem cells. It also induced murine primary calvarial cells to produce a mineralised matrix and caused localised ossification in the epiphysis of an embryonic chick femur. This work has developed a robust, novel method of tailoring protein release and applied it to rhBMP-2. The technology could be transferred to different growth factors and the regeneration of different tissues.

Guanylyl cyclase - cyclic guanosine monophosphate (cGMP) signalling has been demonstrated to play an important role in the endogenous cardioprotective signalling of the myocardium during early reperfusion. It is proposed that infarct limitation is afforded by elevating cGMP and activating protein kinase G and its distal targets. It was hypothesised that increasing the activity of soluble guanylyl cyclase (sGC) would limit myocardial ischaemia-reperfusion injury. Primarily using the rat isolated perfused heart method, the experiments reported in this thesis investigate the role of exogenous targeting of sGC during early reperfusion, specifically exploring targeting different redox states of the enzyme and their effects on myocardial infarct size. The novel sGC stimulator BAY 41-2272 and activator BAY 60-2770 were selected to investigate this hypothesis. Both administration of BAY 41-2272 and BAY 60-2770 during early reperfusion significantly limited infarct size compared to controls. This was associated with elevated total tissue cGMP levels. Inhibition of nitric oxide could not completely abrogate this protection, but exogenous perfusion of nitric oxide along with BAY 41-2272 showed synergistic action. Oxidation of the prosthetic haem group by ODQ abrogated the protection afforded by BAY 41-2272 but potentiated the protection afforded by BAY 60-2770. Targeting both the reduced and oxidised forms of sGC together did not afford additive protection, in fact it reduced the protection afforded compared to the individual treatments. Preliminary data also suggest that targeting the particulate form of guanylyl cyclase increases activity of Akt signalling during early reperfusion suggesting common signalling between soluble and particulate guanylyl cyclase. These data suggest that targeting sGC during early reperfusion can afford cardioprotection by limiting infarct size. The relationship between cGMP elevation and infarct size needs to be investigated further. Nevertheless, these studies suggest that sGC may be a tractable target for the therapeutic management of acute myocardial infarction.

Biodegradable microneedles (MNs) are currently being developed to painlessly facilitate the effective permeation of therapeutic substances across the skin barrier. As sugar glasses are utilised in nature to protect proteins and other delicate structures upon dehydration, such materials may be an appropriate substrate for the preparation of biodegradable MNs. The aim of this work was to investigate for the first time the feasibility of preparing biodegradable MNs from sugar glasses and to test their potential utility for drug delivery applications. Solid sugar products were fabricated from 32 different solutions containing a range of individual sugars and binary sugar combinations, utilising a low temperature dehydration methodology. Subsequently, a novel vacuum-forming micromoulding methodology was developed and optimised to produce sugar glass microneedle (SGMN) arrays from silicon master structures. The sugar materials and MN structures were characterised using a variety of microscopic, thermal and x-ray diffraction analyses. The ability of SGMNs to puncture human skin was assessed in an in vitro skin model, whilst SGMN facilitated drug delivery was investigated using modified static Franz-type diffusion cells. A range of model substances including methylene blue (MB) dye, ibuprofen sodium (IBU), sulforhodamine B (SRB), FITC-BSA and β-galactosidase (β-gal) were incorporated within SGMN arrays. Furthermore, novel SGMN adhesive patches containing SRB within the backing only were fabricated using silicone and acrylate adhesives. Long-term stability of SGMN arrays was assessed under a range of differing storage conditions. Initial characterisation studies suggested that non-crystalline sugar material was formed from anhydrous trehalose and sucrose (75:25 %w/w) sugar solutions. This finding was critical to future SGMN fabrication and incorporation of model substances within the material. Process optimisation led to fabrication of SGMNs with strong morphological fidelity to master structures, which reliably penetrated human skin to facilitate diffusion of MB dye. Furthermore, SGMNs were shown to dissolve rapidly and completely in human skin and deliver MB, IBU, SRB and FITC-BSA to the deeper skin layers. Diffusion study data suggested that SGMN arrays incorporating a range of model substances facilitated permeation across skin in a bolus delivery manner. Additionally, it was found that SGMN adhesive patches were able to control permeation of SRB, a model hydrophilic compound. Sugar glasses containing β-gal were shown to stabilise enzyme functionality at approximately 40 % of initial activity over a 3 month period when stored under desiccation. Elevated humidity and temperature storage was detrimental to SGMN morphology, with 10 % relative humidity at 20 °C being optimal for MN preservation. Overall, this study suggests the utility of SGMNs for the stable incorporation and effective intra- or trans-dermal delivery of a range of model substances, including hydrophilic and macromolecular molecules. Furthermore, it was shown that a novel SGMN adhesive patch may provide the capability to control drug release across skin. Sugar glasses demonstrated a stabilising effect upon a functional protein cargo, although it appeared that storage conditions had a strong influence upon physical SGMN stability.

The aim of the study was to evaluate a community pharmacy-based cardiovascular risk assessment (VRA) service introduced into two pharmacies in south Wales. A longitudinal methodology was adopted where participants had an initial assessment with a follow-up after 12 months. Body mass index, waist circumference, blood pressure and total/HDL cholesterol levels were measured and the Framingham 10-year cardiovascular risk was estimated and communicated to patients. Demographic details and lifestyle information (smoking, alcohol, diet and exercise) were obtained via self-complete questionnaires at each consultation. A total of 172 individuals accessed the service and had either a brief assessment (n=26) without the calculation of the Framingham score or a full VRA (n=146). Mean age was 60 years (±10.3), 59% were female and 25% (37/146) were at high risk (>20%) of developing cardiovascular disease. High satisfaction with the VRA was obtained via an anonymous questionnaire provided immediately after the initial consultation (74% response rate). The short-term outcomes of the service (including recall of advice, lifestyle improvement and/or making the visit to their GP if they were referred) were reported through a semi-structured telephone interview two weeks after the initial assessment. In total 105/172 (61%) who attended the twelve-month follow-up had results of the two assessments compared using paired Student’s t-test. There was a statistically significant increase in mean HDL 0.08 mmol/L (95% CI 0.02 to 0.14) and a statically significant reduction in mean systolic BP -8.5 mmHg (95% CI -11.0 to -5.9), diastolic BP -7.7 mmHg (95% CI -10.4 to -5.0) and Framingham score -1.07 (95% CI -1.9 to -0.2). A comparison between Framingham and QRISK2 algorithms showed the importance of using the most accurate tool available in estimating cardiovascular risk. This is the first study to investigate short- and longer-term outcomes of a community pharmacy-based VRA service in Wales and provides a basis for future research.

Las plantas de uso medicinal son una respuesta del conocimiento ancestral para el tratamiento y/o cura de diversas enfermedades, que debe ser contrastado científicamente. Dicha investigación debe partir de la identificación botánica de los ejemplares utilizados por la población; asimismo, el conocimiento de sus metabolitos nos ayuda a explicar el o los principios activos implicados en la actividad atribuida a la planta. El material empleado en este estudio son tres helechos comercializados bajo el nombre común de “cuti cuti” en el mercado de plantas medicinales del distrito de La Victoria en Lima-Perú, cuya identidad taxonómica corresponde a las especies: Argyrochosma nivea (Poir.) Windham “cuti cuti hembra”, Cheilanthes pruinata Kaulf. “cuti cuti macho” y Cheilanthes scariosa (Sw.) C. Presl “cuti cuti”, los cuales son empleados en la medicina tradicional para tratar la diabetes. Se ha podido establecer características descriptivas que conlleva a la presentación de una clave que permite diferenciarlos. Se demuestra que los helechos llamados “cuti cuti” son de tres especies distintas. Se procedió a realizar ensayos de colorimetría y precipitación, a partir de extractos hidroalcohólicos resolubilizados en agua al 2%, encontrándose similitudes en los tres helechos respecto a la presencia de metabolitos.

Mini-dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy, Durban Institute of Technology, 2002. / The first objective was to investigate the effect of Sutherlandia frutescens 30CH on healthy provers and to record the signs and symptoms produced, so that it could be prescribed according to the Law of Similars, as required by homoeopathy. The second objective was to analyze the symptoms obtained from this proving according to the doctrine of signatures. It was hypothesized that Sutherlandia frutescens 30CH would produce observable signs and symptoms in healthy volunteers which would correlate to a doctrine of signatures analysis of the plant. A double blind proving of Sutherlandia frutescens 30CH was conducted. 24 provers were selected and randomly divided into two groups, the first consisted of 18 provers who received medicated powders and the second consisted of the remaining six who received placebo. The subjects were unaware of the nature of the substance that they took nor the potency thereof as an added control measure. Provers were examined and kept journals before, during and after administration of the remedy so as to serve as their own control. This information, along with data collected by the researchers from case histories and physical examination, was then assessed on completion of the proving. The researchers then translated the symptoms elicited into Materia Medica and repertory language and formulated a homoeopathic picture of this remedy. Data was analyzed by qualitative methods, as the data was not amenable to standard / M

Submitted in fulfillment of the requirements for the Degree of Masters: Homoeopathy, Durban Institute of Technology, 2007. / The aim of this study was to evaluate the therapeutic potential of an indigenous South African substance and the traditional uses of that crude substance. The substance under evaluation was Erythrina lysistemon which was prepared homoeopathically to the thirtieth centesimal (30CH) potency. / M

Mini-dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy, Durban Institute of Technology, 2002. / The purpose of this investigation was to determine the effect of Sutherlandia frutescens 30CH on healthy provers, and to record the signs and symptoms produced, so that it may be prescribed according to the Law of Similars. A further aim of the investigation was to compare those remedies yielding the highest numerical value and total number of rubrics on repertorisation of the proving symptoms to Sutherlandia frutescens. It was hypothesised that the 30CH potency of Sutherlandia frutescens would produce clearly observable signs and symptoms in healthy provers, and that the comparison of Sutherlandia frutescens to those remedies yielding the highest numerical value and total number of rubrics on repertorisation of the proving symptoms would highlight differences and similarities between the remedy symptoms so that confusion as to indication is erased. It was hypothesised that a fuller understanding of Sutherlandia frutescens and its relationship to other remedies would be gained following this comparison. III A double blind, placebo controlled proving of Sutherlandia frutescens 30CH was conducted on twenty-four healthy volunteers who met the inclusion criteria. Six of the twenty-four provers randomly received placebo, with neither prover nor researcher knowing who was on placebo. Provers had a homoeopathic case history taken and physical examination performed on them before commencement of the proving. The provers recorded their signs and symptoms before, during and after administration of the remedy, by means of a journal. On completion of the proving, the information was correlated and assessed by the four researchers, the symptoms elicited translated into Materia Medica and Repertory language, and a homoeopathic picture of the remedy formulated. Data from the case histories, physical examinations and group discussion were also considered. / M

Mini-dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy, Durban Institute of Technology, 2004. / The first objective was to investigate the effect of Sutherlandia frutescens 30CH on healthy provers and to record the signs and symptoms produced, so that it could be prescribed according to the Law of Similars, as required by homoeopathy. The second objective was to analyze the symptoms obtained from this proving in a direct comparison to the effects of the major pharmacologically active compounds present in Sutherlandia frutescens. A double blind proving of Sutherlandia frutescens 30CH was conducted. Twenty-four provers were selected and randomly divided into two groups, those receiving medicated powders (18 subjects) and those receiving the placebo powders (6 subjects). As an added control measure, the subjects were also ignorant to both the nature of the proving substance and the administered potency. Prior to taking the remedy, each patient provided their own case history and received a physical examination so as to establish each individual's baseline. Each prover was then required to keep a daily journal, in which all symptoms were recorded in accordance with a suggested guide. The researchers then collated the data and translated the symptoms produced into Materia Medica and repertory language. Finally a homoeopathic picture of the remedy emerged in which marked themes exist. These themes and central characteristics of the remedy were then compared to the toxicology of the major chemical constituents of Sutherlandia frutescens. Data was analyzed by / M

Mini-dissertation submitted in partial compliance with the requirements for the Master's Degree in Technology: Homoeopathy, Durban Institute of Technology, Durban, 2002. / The purpose of this study was to determine the effects of Sutherlandiafrutescens 30 CH on healthy individuals so that it may be prescribed according to the law of similars. It was also the purpose of this study to compare all existing indications of use of Sutherlandia frutescens to the proving symptomatology. This would allow a greater understanding of the plants spheres of action and would contribute to the formation of a comprehensive remedy picture by highlighting similarities and differences between the homoeopathic and traditional indications of use. The comparison also served to investigate whether Sutherlandia frutescens acts, in accordance with the fundamental law of homoeopathy, the law of similars, in traditional application. It was the intention of the study to shed light on the possible 'homoeopathicity', or not, of current traditional use of Sutherlandia frutescens, as opposed to overt pharmacology of phytochemicals. It was hypothesized that Sutherlandiafrutescens 30CH would produce clearly observable signs and symptoms in healthy provers. The second hypothesis was that the proving symptomatology of Sutherlandia frutescens 30CH would be similar to the existing indications of use. The homoeopathic proving was a double blind placebo controlled study conducted by four Master's in Technology: Homoeopathy students. A total of 24 subjects formed the proving group, 25% of whom (6 of the 24) were randomly assigned to the placebo group. The subjects were unaware of the substance they were proving and the potency of the substance to be proven. The provers also served as intra-individual controls by recording their state prior to the administration of the remedy, to provide a baseline for comparison after the administration of the remedy. Provers took one powder three times daily until proving symptoms appeared, but for no longer than 2 days i.e.: a maximum of 6 doses. Provers recorded their symptoms daily in a journal and were in / M