The tropical rainforest of Malaysia is considered as one of the most evolved and complex ecosystems in the world that serves a vast untapped biodiversity of natural resources. Exploitation of medicinal plants for bioactive compounds is of great potential and could be an imperative source of providing new vistas for novel drug discovery and development. The study was undertaken to evaluate the in vitro pharmacological properties of Artabotrys crassifolius including antibacterial, antifungal, anticancer and antioxidant activities of the plant. The leaves and bark of Artabotrys crassifolius were extracted sequentially using hexane, chloroform and ethanol. The prepared crude extracts were subjected to phytochemical screenings for the presence of alkaloids, cardiac glycosides, flavonoids, phenolic compounds, saponins, tannins and terpenoids. Kirby-Bauer disc diffusion assay was conducted to examine the antibacterial and antifungal activities of crude extracts against ATCC and clinical strains. The anticancer effect of crude extracts was investigated against human breast and colorectal carcinoma cell lines using MTT assay whereas the antioxidant potential of crude extracts was assessed using TPC, TFC, ABTS, DPPH and FRAP assays. Among the crude extracts studied, hexane and chloroform extracts of bark exhibited pronounced antibacterial activities against ATCC and clinical strains with zones of inhibition ranging from 8.23±0.25 mm to 13.70±0.26 mm and 7.75±0.25 mm to 13.68±0.28 mm respectively. However, all the crude extracts were found to be devoid of antifungal activity except for hexane extract of bark which was able to inhibit the growth of the tested Candida species with zones of inhibition ranging from 7.81±0.27 mm to 9.77±0.25 mm. In addition, chloroform extract of bark was highly active against all of the tested carcinoma cell lines with GI50 values ranging from 4.23 µg/mL to 9.45 µg/mL, while hexane extract of bark potently inhibited the growth of MDA-468 breast and HCT-116 colorectal carcinoma cell lines with respective GI50 values of 6.10 µg/mL and 16.45 µg/mL. Furthermore, ethanol extract of bark that possessed the highest total phenolic and flavonoid contents (268.29±12.36 mg GAE/g and 179.54±4.98 mg CE/g) was shown to demonstrate prominent scavenging activities against ABTS cation and DPPH radicals with IC50 values of 16.50 µg/mL and 16.54 µg/mL respectively, as well as exceptionally high antioxidant power with FRAP value of 1884.35±83.78 µmol Fe(II)/g. The chromatographic separation of chloroform extract of bark led to the isolation of four alkaloids, namely artabotrine, liridine, atherospermidine and lysicamine. Among the compounds isolated, artabotrine displayed high antibacterial properties with respective MIC and MBC values ranging from 1.25 µg/mL to 5 µg/mL and 1.25 µg/mL to 20 µg/mL against all of the tested ATCC and clinical bacterial strains, with the exception of Actinobacillus sp. and Klebsiella sp. Moreover, artabotrine was highly active in HCT-116 colorectal and MCF-7 breast carcinoma cell lines with GI50 values of 3.34 μM and 3.49 μM respectively. In conclusion, exploration of the in vitro pharmacological properties of Artabotrys crassifolius revealed that artabotrine with dual antibacterial and anticancer activities may represent a new generation of potential drug candidates for the treatment of bacterial infections and cancer. Hence, further in vivo studies and clinical trials are required to ascertain the efficacy, safety and mechanisms of action of artabotrine prior to application in the pharmaceutical industry as natural therapeutic agents.

Pharmacy as a profession is changing rapidly in the UK. Over recent years, the increased utilization of ready-prepared drugs has led to a decline in the need for the traditional skills of formulation, while computerization has resulted in a situation where much of the routine dispensing work can be undertaken by less qualified personnel. The decline in the traditional aspects of pharmacy has been matched by the emergence of a much greater advisory role. Pharmacy practice researchers have been drawn to support these developments by investigating related areas, but the common factor linking this research is its focus on clinical as opposed to communication issues. Rather than investigating the nature of face-to-face interaction between pharmacists and clients as a topic in itself, researchers instead have been largely concerned with patient/health care system mteractions as a function of drug therapy. Those few studies that have focused exclusively on communication have done so from a quantitative, social psychology framework, thus ignoring the two way, reactive nature of the interaction process. This study, using data collected from patients' and carers' consultations with pharmacists in a hospital paediatric oncology outpatient clinic, uses the sociological methodology of Conversation Analysis (CA) in order to analyze the encounters which take place. In so doing, it aims to shed some light upon what is actually involved in the process of "patient counselling" in this setting. The body of CA literature which considers advice-giving in health care settings provides the starting point for a consideration of the ways in which pharmacists give advice in this setting, and how this is responded to. The aims are thus twofold: to enlarge the methodological resources of PPR, and also to begin an examination of the communicative competencies required of pharmacists in this setting. NB. This ethesis has been created by scanning the typescript original and may contain inaccuracies. In case of difficulty, please refer to the original text.

The cannabinoid receptors CB1 and CB2, which belong to the rhodopsin family of GPCRs, are implicated in the pathology of various disease states. As drugs targeting these receptors remain limited, novel cannabinoid receptor ligands represent an unmet need with substantial therapeutic potential. We present here the construction and application of homology models of the human CB1 and CB2 cannabinoid receptors based on the crystal structure of the human adenosine A2A receptor for the structure-based design of novel ligands based on the fenofibrate scaffold. Models were refined through molecular dynamic simulations in a lipid bilayer, and were validated via the prediction of known ligand binding affinities, enrichment studies and assessment of predicted ligand binding modes. These validated models were subsequently used in predicting the binding mode of fenofibrate derivatives to the cannabinoid receptors. The predicted binding mode of these fenofibrate derivatives to the CB2 receptor showed good agreement with known mutagenesis data, indicating the binding of these compounds to be stabilized primarily by hydrogen bonds with W5.43 and C7.42, aromatic stacking with F2.57, F3.36 and W6.48, and hydrophobic contact with F2.64, V3.32 and I5.47. A series of novel ligands was derived based on these findings, docked into our model, synthesized and pharmacologically evaluated at the CB2 receptor. The pharmacology of these ligands validated our modelling predictions and binding mode hypothesis, with several of these ligands showing unique pharmacology by binding in an allosteric manner. These findings may be used to guide the design of further derivatives and highlight the promise of the fenofibrate scaffold in the development of novel CB2 receptor ligands.

Media promotion of the ideal body as slimness for women and muscularity for men, has led to increasing numbers of both genders reporting dissatisfaction with their bodies and trying to change using weight control products. It has been suggested that pharmacists can play a key role in promoting healthy lifestyles and weight management. The main aim of the research study was to examine the impact of media on body image perception and to investigate the role of pharmacists in weight management. This thesis consists of three studies: an evaluation of weight control websites, a body image survey of young adults and interviews with pharmacists. The results from the evaluation study showed that the quality of most weight loss (64%) and weight gain product advertisements (80%) was generally poor, principally due to the use of misleading claims and a lack of useful information. These data complement the survey that university students reported rarely being influenced by weight control product adverts and hardly considered using these products as a method for changing weight. The survey also showed that most participants of both genders (71%) felt satisfied with their body image. However, the findings in this study indicated that there was a relationship between a high level of body image concern and self-perception of being overweight and the attempt to lose weight. Interviews with pharmacists showed that they were unlikely to be actively involved in reducing obesity problems and promoting healthy lifestyles in the community. Although the prevalence of body dissatisfaction and the use of weight control products among these young adults may not be as high as has been reported elsewhere, healthcare professionals, including pharmacists, need to be aware of this problem when providing advice or consultations for young adults. Healthcare professionals should promote the benefits of healthy lifestyles as well as providing information about potential problems of harmful weight control strategies. The effect of ethnicity and culture on body image concern warrants further. The availability of weight control products and quality of information provided should also be further examined.

The aim of this project was to formulate a solid core drug delivery system for oral delivery of proteins using silica as core material, haemoglobin as model drug and supercritical fluid processing as encapsulation technique. Silica particles of different morphology were used as a carrier material for protein immobilisation and fatty acids coating was performed using supercritical carbon dioxide (CO2) as a processing media. The melting behaviour of saturated fatty acids (lauric, myristic, palmitic and stearic acid) and pluronics (F-38, F-68, F-77, F-127 and F-108) were studied under pressurised CO2 to identify the coating parameters. These excipients showed a melting point depression in the range of 10 to 20 °C in pressurised CO2. In the case of fatty acids the decrease in melting point was inversely proportional to the carbon chain length and directly related to the polarity of carbonyl group. Whereas, melting point depression for all pluronics was similar and was attributed to the high cohesive energy density of these polymers. This phenomenon was used to encapsulate the thermolabile protein molecules in pressurised CO2 at low temperatures. The stability of bovine haemoglobin (bHb) was studied by ultraviolet-visible and circular dichroism spectroscopy. Thermal, storage and agitation stability were studied to identify the processing parameters for the storage, adsorption and desorption processes. Three morphologically different silica particles were studied as potential inorganic carrier for the protein. The particles were characterised by nitrogen adsorption and scanning electron microscopy and the maximum protein adsorption and kinetics was determined at pH 6. The bHb adsorption on silica was found to be irreversible, hence application of pluronics as a displacer was also studied. Finally, the bHb adsorbed particles were coated with fatty acids by supercritical fluid processing and solvent evaporation methods. The highest bHb release was obtained from SFP (Syloid FP-244) silica in comparison to other silica particles in pH 6.8 phosphate buffer. SFP based formulation also showed a trend in the protein release which was dependent on the solubility of fatty acids in the release media. The highest release was obtained from myristic acid coated solid core drug delivery system (SCDDS) followed by palmitic and stearic acid. Lauric acid coated SFP formulations led to changes in protein conformations, hence omitted from these studies. The release studies of myristic acid based SCDDS in simulated gastric and intestinal fluids showed that fatty acid coating provided enteric properties to the formulation. It can be concluded from these studies that SCDDS prepared using mesoporous silica as core and fatty acids as coating material can be an effective drug delivery system for the oral delivery of biomolecules.

En el mundo tan complejo en el que nos movemos, sería muy fácil reducir las cuestiones éticas de un médico a grandes temas como el aborto o la eutanasia. Sin embargo cada acto médico, cada interacción con un paciente tiene un gran componente ético. Esa ética del día a día que conlleva no solo la empatía e incluso la compasión sino tambien una buena comunicación. En la charla Ética Médica a Colores la Dra. Lalanda hace una llamada sobre las cosas del día a día en un centro sanitario, todas esas que no llegan a las portadas de los periódicos pero pueden hacer de un buen médico un médico bueno.

This work describes the creation of a three-dimensional model of the children’s brain tumour medulloblastoma using primarily human cells. This in vitro cell culture model was created as a platform for testing novel drug delivery systems for local delivery in the brain. The aim of the local delivery strategy was to reduce radiotherapy through the use of nanoparticle-based chemotherapy. The nanoparticles would be delivered after surgery in the cavity left by the excised tumour tissue. The model was intended to evaluate the selective cytotoxicity of advanced drug delivery systems towards tumour tissue and the benefit of nanoparticle therapy compared to free drug. Normal tissue was modelled using human foetal brain tissue and tumour tissue was represented by a variety of medulloblastoma cell lines. Both were cultured as three-dimensional spheroids free of artificial matrix in ultra-low attachment plates. The tumour and normal cells could be cultured either separately or together and the viability for each cell population determined using a battery of methods. Co-cultures of both cell types had the additional benefit of mimicking the interaction between normal and tumour tissue. The use of physiologically relevant single and co-culture in vitro models could provide information on the relative safety and efficacy of novel brain tumour treatments. The high-throughput platforms used, the algorithms and the validation of a battery of tests in 3D may be extrapolated to other cancer models as well. Moreover the universal marking procedure employed can be employed to label, culture and analyse any two cell types, while preserving tissue heterogeneity and viability. The key benefit from this thesis is the framework for designing in vitro models of tumours that include normal tissue as an internal control. This is an important contribution that can substantiate IC50 values by putting them in the context of drug safety and efficacy. It also highlights the minimum checks and feasibility experiments that need to be done before an in vitro assay is accepted for 3D spheroids.

Clusianone isolated from Garcinia parvifolia has been studied for its anticancer properties and cytotoxicity to human respiratory cells. The clusianone was first isolated using solvent extraction, column chromatography, thin layer chromatography and finally recrystallization method through solvent evaporation including seed crystal to induce crystal growth of clusianone. The clusianone was characterized using X-ray crystallography, ESI-MS, NMR and melting point. During the course of this research, clusianone was chemically modified and eight different derivatives abbreviated as CMet, CHyd, CMxA, CMeA, CEtA, CPryl, CGeryl and CDMet were obtained. Some of the chemical methods employed were hydrogenation, methylation, demethylation, ketone reduction via addition of amine derivatives and O-alkylation to install additional prenyl and geranyl chain into clusianone. Further studies of the role of the clusianone derivatives were presented in its in vitro anticancer activity. The anticancer test and cytotoxicity effect were tested using MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)assay. Specific normal human and cancer cell lines were used in the assay being MRC5 (lung fibroblast), A549 (lung adenocarcinoma), NP69 (nasopharyngeal epithelial cell) and HK1 (squamous carcinoma of the nasopharynx). Preliminary study of clusianone and derivatives showed cytotoxicity effect in dose dependent manner. Based on the results, clusianone and compound CMeA demonstrated good anticancer activity showing IC50 values below 5 μM against A549 and HK1 cancer cells and at the same time affecting less of the MRC5 and NP69 normal cells. Western blot method was employed to further elucidate the downregulation of the protein expression of β-tubulin including cell cycle regulators Cdk1 and cyclin B1 for clusianone and CMeA derivative treated carcinoma cells. Clusianone and compound CMeA demonstrated potential antimicrotubule agent characteristic since expression of β-tubulin and cell cycle regulators Cdk1/cyclin B1 were downregulated in A549 cells. As for HK1 cells, clusianone downregulated β-tubulin protein without affecting Cdk1 and cyclin B1 expression. In contrast, compound CMeA showed prominent downregulation of β-tubulin, CDK1 and cyclin B1 proteins especially after 48 hours treatment in HK1 cells.

Apoptin, a potential anticancer candidate, selectively kills tumour or transformed cells but remains harmless to normal and non-transformed cells. Besides, apoptin-induced apoptosis is independent of p53 apoptosis pathway, which is always mutated in cancer cells during tumorigenesis or after radiotherapy. This has made apoptin becoming more pharmacological valuable. In this study, the general aim was to develop a plant-derived apoptin which offers a safer and more cost-effective treatment for cancer patients. The scope of the study focused on the production of recombinant apoptin in a plant-based system and preliminary bioactivity evaluation of the purified plant-derived apoptin on human lung cancer adenocarcinoma A549 cells. Recombinant apoptin was expressed in Nicotiana benthamiana as apoptin alone, in fusion to green fluorescent protein (GFP) as well as in fusion to lichenase (Lic) to increase the expression of recombinant protein in soluble fraction. Recombinant apoptin was also in fusion to H22 single chain antibody and epidermal growth factor (EGF) in order to target recombinant H22-apoptin and EGF-apoptin to cancer cells overexpressed with immunoglobulin G (IgG) receptor (CD64) and EGF receptors. Expression of recombinant apoptin was detected in N. benthamiana successfully, however, high amount of soluble protein was obtained in plants infiltrated with recombinant GFP-apoptin (gene casette: PR-GFP-VP3-HK) and EGF-apoptin (gene casette: PR-EGF-CatAd-VP3-HK) that targeted the recombinant proteins to endoplasmic reticulum (ER). Protein purification using immobilised metal affinity chromatography (IMAC) recovered recombinant GFP-apoptin (GFP-VP3-HK) and EGF-apoptin (EGF-CatAd-VP3-HK) at a low purity when recombinant proteins were purified in a non-denaturing condition. Host cell protein contamination was not able to be removed when second chromatography and acid precipitation method were used. However, recombinant GFP-apoptin (GFP-VP3-H) and lichenase-apoptin (Lic-VP3-H) without targeted to specific cellular compartment were purified at a good purity using IMAC. Recombinant GFP-VP3-H extracted in a denaturing condition was successfully refolded without an addition of chemical additives while recombinant Lic-VP3-H required triton to refold the protein. In cell-based study, enzyme-linked immunosorbent assay (ELISA) showed that apoptin interacted with EGF receptors as well as A549 cells which finding is the first of its kind in report but with an unverified speculation. On the other hand, nuclear localisation activity and a few apoptosis-associated morphological features were observed in cells microinjected with recombinant GFP-VP3-H. Meanwhile, recombinant EGF-VP3-HK showed a dose-dependent growth inhibitory effect at higher concentration and caused the loss of mitochondrial membrane potential (MMP) in treated A549 cells. However, internalisation of recombinant EGF-VP3-HK requires a further study for confirmation. Considering the findings on MMP and caspase 3/7 assays were not convincing enough, further evaluations are necessitated in order to verify the apoptosis events induced by the recombinant GFP-VP3-H and EGF-VP3-HK. Nonetheless, this study has heralded a new milestone for apoptin research by which its protein has been successfully produced in plants and some preliminary biological characteristics have been explored at a certain extent.

Computerised clinical decision support systems (CDSS) are information technology tools used to optimise the use of antibiotics. A systematic review and meta-analysis was conducted to review the level of evidence of the impact of CDSS on antibiotic prescribing using specific outcome measures. Overall, CDSS interventions were associated with an increase in adequacy of antibiotic coverage based on a random effects model [OR = 2.11, 95% CI, 1.67 to 2.66, p ˂ 0.00001]. Results showed that CDSS had a marginal statistically significant effect on mortality based on a random effects model. [OR = 0.85, 95% CI, 0.75 to 0.96, p = 0.01]. Medical and non-medical healthcare professionals in University Hospitals Birmingham Foundation Trust were surveyed about their perceptions and attitudes towards CDSS. 85% participants showed a positive attitude towards CDSS. A quantitative retrospective before-and-after study in the University Hospitals Birmingham Foundation Trust was conducted from June 2012 to June 2016 to measure the impact of a CDSS tool known as Structured Prescribing on the volume of antibiotic use. From June 2012 to June 2016, the total antibiotic usage increased by 13.1% from 1436.3 to 1624.85 DDD/1000 bed-days. CDSS show demonstrable potential in optimising the use of antibiotics and containing antimicrobial resistance.