Background: The Royal Stoke Pharmacy Workforce Calculator (RSPWC) was developed to meet a local need of identifying clinical pharmacy staffing levels, not described nationally. This study demonstrates the validity and transferability of the RSPWC and its application to other settings. Methods: A two-round Delphi consensus study was conducted to confirm the activity standard (tasks, times and frequencies) for clinical pharmacy services. An operator evaluation was undertaken to demonstrate the reliability of the tool by multiple operators and a series of qualitative interviews explored the utility of the tool in different settings. These research strands ran concurrently from April 2016 to December 2016. Results: Participants from 21 sites across the UK were recruited, including district general hospitals (nine), teaching hospitals (eleven) and one mental health trust. A wide range of staffing levels, across all staff groups, was reported. Consensus was achieved for 68% of components of the algorithm that drives the RSPWC. For a further 21% of components, ‘nationally-representative’ figures were identified from the data. Eleven percent of the components, those relating to elements dependent on individual patient responses to medicines, failed to achieve agreement. A pragmatic approach was taken in the derivation, from study data, of these activity frequencies for typical patient groups. Content validity of the tool was demonstrated. The ‘operator evaluation’ demonstrated reliability in its use by different operators. The application of this tool to a variety of settings was identified through the qualitative data. Discussion: The results of the study demonstrate the validity, transferability and utility of the RSPWC. They capture, for the first time, a consensus on the required service components for the delivery of pharmaceutical care, across multiple hospital sites nationally in the UK. Through this study a clinical pharmacy workforce calculator for acute hospital settings has been developed and validated.

Background: Anticoagulation therapy for patients with atrial fibrillation (AF) remains a national challenge. A decision support tool (DST) was developed to assist healthcare professionals (HCPs) in the appropriate prescribing of anticoagulants in patients with AF. This thesis aimed to evaluate the utility of the DST and associated patient decision aid (PDA) for anticoagulant decision making in clinical practice. Methods: This study involved a series of sequential stages in the evaluation of the DST. Semi-structured interviews were conducted with forty-seven HCPs to explore their perceptions of anticoagulation prescribing decision. Using a vignette, the perspective of HCPs on the potential utility of the DST and associated PDA were explored using both semi-structured interviews and questionnaires. Second interviews were conducted approximately eight weeks from the initial contact to explore HCPs’ perspectives on the actual utility from implementing the DST and associated PDA in routine clinical practice. The perspectives of a group of AF patients’ who had experienced the DA during consultation were explored using semi-structured interviews and questionnaires. Results: Qualitative themes elicited during initial contact revealed that anticoagulants prescribing decision can be suboptimal. Findings from the pre-intervention evaluation showed that the DST has potential to improve the quality of anticoagulants decision process. Findings from post-intervention evaluation demonstrated improvements in anticoagulants decision-making in clinical practice. Findings from fourteen patients revealed that the DA was effective in facilitating a quality decision that was informed and consistent with personal values and expectations. Conclusions: This study demonstrated the positive impact the DST can have on the quality of anticoagulants decision-making in clinical practice and provides a unique contribution to the existing CDSS research. The ever-increasing demand for a quality decision-making process in clinical practice is a fertile environment for clinicians and policymakers to consider the potential impact that the DST and associated PDA can offer.

Virtual patients (VPs) are routinely used in the training of medicine and nursing professionals but uptake into pharmacy has been slower. The pharmacy pre-registration training year takes place in the workplace and a disparity in the perceptions of support provided and the pre-registration examination pass rates has been established between the training sectors. This programme of work aimed to evaluate the effectiveness of virtual patients (VPs) at supporting pre-registration training when compared to a non-interactive (NI) learning tool. Following institutional ethical approval, a mixed methods approach was adopted to evaluate the VP technology. A purposive sample of 165 pre-registration trainees (2014-2015) who were completing their training in a UK-based community or hospital pharmacy were recruited. Participants were randomly stratified to receive three VP or NI case studies. Knowledge surrounding the case studies was assessed using a quasi-experimental evaluation and thoughts on the two learning tools were obtained and compared via questionnaires and semi-structured telephone interviews. Quantitative data was analysed using descriptive and inferential statistics and qualitative data was analysed using content analysis (questionnaire) and framework analysis (interviews).No significant differences in knowledge improvement between pre-registration trainees in the VP and NI groups were obtained. Significant improvements in knowledge were found between the sectors of training for the three case studies. Pre-registration trainees reported that the VP enabled them to apply their learning and engage in experiential learning. The VP case studies were associated with greater satisfaction and were reported to provide a more realistic, interactive and enjoyable learning experience. Pre-registration trainee’s perspectives of the VP technology as a learning tool were more favourable regarding the development of real-life complex skills and aspects of learning, which provides a remit for further evaluation of the technology in undergraduate and postgraduate pharmacy training.

Hepatocellular carcinoma accounts for 85% of primary liver cancer that are usually characterised by defective or ineffective apoptosis which is considered to be the main cause of cancer progression. In this study, Cytochrome-C which is a pro-apoptotic protein is combined with hybrid (iron oxide-gold) nanoparticles and triggers mitochondrial downstream apoptosis pathway in the tumour cells. The nanoparticle complex enables delivery of this difficult protein through the cell membrane. In this research, five different anticancer drugs (doxorubicin, oxaliplatin, paclitaxel, vincristine and vinblastine) were used against liver cancer and U937 cell lines to assess their IC50 values alone and to check their toxicity after their co-administration with cytochrome C hybrid formulation. These combinations resulted in an increase in the cytotoxicity of the used chemotherapeutic drugs and remarkable decrease in the amount needed to kill hepatic cancer cells. For that reason, Iron-gold hybrid nanoparticles offer a promising tool for cytochrome-c delivery into tumour cells and enhance the specific targeting of therapeutic particles to their site of action. The preliminary results reflected the increasing killing abilities of chemotherapeutic therapies when co-administered with cytochrome C hybrid formulation by targeting the natural killing mechanism inside the cells and activating its pathways. Subsequent to these results, further work was done in formulation of one platform therapeutic device with Polymeric amphiphiles hydrophilic poly(allylamine) polymer (PAA) grafting with 5-(4-chlorophenyl)-1,3,4-oxadiazole-2-thiol(oxadiazole, Ox). Paclitaxel (PTX) was selected as a hydrophobic drug model to check its water solubility behaviour after loading into PAA-HNP-C platform. These new devices showed a significant increase in drug uptake level and increase in PTX cytotoxicity against liver cancer cell lines. The data from this work showed a significant increase in the apoptosis activities of combining treatment anticancer agents (doxorubicin, paclitaxel, oxaliplatin, vinblastine and vincristine) and the hybrid formulation of the cytochrome-C within the liver cell lines, which leads to cellular death. Therefore, this combined method may give promise step for the future of liver cancer treatment regimes. In addition to, formulating the HNP-CYT-C and PTX into as active single platform for increasing the PTX cytotoxicity. More laboratory investigation is needed to check the activity of this formulation as a preparing step to further in vivo studies.

A estabilidade de agregados é influenciada pelos sistemas de manejo do solo, com reflexo na estabilização da matéria orgânica e estoques de C e N no solo. O efeito de longo duração (30 anos) de dois sistemas de preparo [plantio direto (PD) e preparo convencional (PC)] e de três sistemas de culturas: [aveia/milho (A/M), ervilhaca/milho (V/M) e aveia+ ervilhaca/milho+ caupi (A+V/M+C)] sobre os índices de agregação, conteúdo e frações de C e N de um Argissolo Vermelho distrófico tipico, foram avaliados. Amostras de solo foram coletadas nas camadas de 0-5, 5-10 e 10-20 cm e separadas em seis classes de agregados (>4,76; 4,76-2,00; 2,00-0,50; 0,50-0,250, 0,250-0,053 e <0,053 mm) por via úmida e seca, e avaliadas em relação ao teor de C e N e submetidas ao fracionamento granulométrico da matéria orgânica do solo (MOS). As concentrações de C e N no solo foram maiores no solo em PD e na camada de 0-5 cm. Os estoques de C e N foram 23 e 32% maiores respectivamente no sistema de culturas (0-20 cm) com elevado aporte de resíduos (A+V/M+C), em relação ao sistema com gramíneas (A/M). O diâmetro médio ponderado (DMP) e o índice de estabilidade dos agregados do solo [ (DMPu/DMPs) x100], foram superiores no solo em PD em relação ao solo em PC, devido ao predomínio de macroagregados (≥ 4,76-2,00 mm). Nos estoques das frações da MOS, o carbono orgânico particulado (COP) e o nitrogênio particulado (NP) foram maiores nos sistemas de culturas com leguminosas no solo em PD, em relação ao PC, enquanto que o C e o N associado aos minerais foram influenciados somente pelos sistemas de culturas. As taxas anuais de acúmulo de C na camada de 0-20 cm nos sistemas de culturas variaram de 35,3 a 67,6 kg C ha-1 ano-1, o acúmulo de N variou de 4,7 a 16,7 kg N ha-1 ano-1 sendo maiores no sistema A+V/M+C. Os estoques de C e N no solo foram influenciados em maior grau pelo sistema de culturas do que pelo sistema de preparo de solo. Esses resultados demonstram que em sistemas conservacionistas o aumento nos estoques de C e N do solo e a estabilidade dos agregados estão associados tanto com a não mobilização do solo quanto com o aporte de resíduos vegetais pelos sistemas de culturas. / The aggregate stability is influenced by soil management systems, with reflex on soil organic matter stabilization and C and N stocks. We evaluated the long term (30-yr) effect of two tillage systems [no-tillage (NT) and conventional tillage (CT) ] and three cropping systems [oat/maize (O/M), vetch/maize (V/M), and oat + vetch/maize + cowpea (O+V/M+C)] on stability of soil aggregates, C and N content in total soil organic matter (OM) and particulate and mineral-associated fractions of a sandy clay loam Typic Paleudult. Soil samples were collected at 0-5, 5-10 and 10-20 cm. Disturbed soil samples were collected for C and N analysis and OM fractionation; undisturbed soil samples were collected for soil aggregates analysis. Aggregates were separated in six classes (> 4.76; 4.76-2.00; 2.00-0.50; 0.50-0.250, 0.250- 0.053 and <0.053 mm) by dry (d) and wet (w) methods, the mean weighted diameter (MWD) of aggregates and the stability index [(MWDw/MWDd)x100] were calculated. ). The concentrations of C and N in soil were higher in NT in 0-5 cm layer. The C and N stocks were 23 and 32% higher in cropping system (0-20 cm) with a high crop residue input (O+V/M+C) than of grasses only based cropping system (O/M). The average diameter and stability index of soil aggregates were higher in NT in relation to CT, mainly due to predominance of macroaggregates (≥ 4.76 to 2.00 mm). In SOM fractions, particulate organic carbon (POC) and particulate nitrogen (NP) were higher in legume-based cropping systems in NT, related to CT, while the C and N in mineral-associated soil organic matter was affected by only cropping systems. The annual rates of C accumulation at 0-20 cm in cropping systems ranged from 35.3 to 67.6 kg ha-1 yr-1, N accumulation ranged from 4.7 to 16.7 kg ha-1 yr-1 and it was higher in the system O+V/M+C. Soil C and N stocks were influenced mainly by cropping systems than the tillage systems. Our results highlight that increase of soil C and N stocks are related both by no soil disturbance plus higher C input under conservationist soil management systems.

Thermo-responsive polymers are a class of smart polymers that respond to change in temperature. This property makes this type of polymers useful materials in a wide range of applications especially, in the field of drug delivery system. Polymers, which have primary amino groups, such as N-(3-aminopropyl) methacrylamide hydrochloride (APMA), have been used for post-polymerization modification reactions for the development of new materials for biomedical applications. Here I aim to fabricate an amphiphilic monomers composed of APMA by substituting (palmitoyl, dansyl, cholesteryl and oxadiazole) groups onto the primary amine in the APMA monomer. These monomers were then copolymerized with N-(2-hydroxypropyl)methacrylamide (HPMA) in six different monomer feed ratios in order to characterise the effect of hydrophobic pendants on the copolymer properties including lower critical solution temperature (LCST), solubility, drug loading and drug release. In this study drug loading and release properties of HPMA-co-APMA copolymers were studied by using four model hydrophobic drugs, propofol, griseofulvin, prednisolone and paclitaxel. High performance liquid chromatography (HPLC) measurements were performed in order to compare drug loading properties of the copolymer formulas. The most potent carrier candidate was loaded in order to carry out thermo-responsive release study. The results showed that all the copolymer formulations in this study possessed the ability to encapsulate practically poor-soluble drugs within their hydrophobic core. HPLC measurement has demonstrated that HPMA-co-(APMA-Oxadiazole 1%) (O-1) and HPMA-co-(APMA-Dansyl 2%) (D-2) have a higher drug solubilisation capacity than other copolymer formulations. In vitro release profiles of different model drugs of the optimal formulation of copolymers were investigated at four different temperatures. Unfortunately, these copolymers showed uncontrolled release of all of the loaded drugs. To set-up thermo-responsive copolymers for controlled drug release, approaches based on introducing poly ethylene glycol (PEG) block as a part of O-1 and D-2 amphiphilic copolymers. A significant enhancement in response to the change in temperature as the drug release across the membrane was seen when PEG was present for the three hydrophobic drugs models. Then hybrid nanoparticles (HNPs) were prepared and attached to O-1 as localised nano-heaters to accelerate drug release in responded to temperature change. Finally, the optimal formulation of (O-1)-b-(PEG) and (O-1)-b-(PEG)-HNPs loaded with paclitaxel (PTX) were tested for their cytotoxicity in vitro on BxPC-3 cells. In-vitro MTT assay results established the ability of (O-1)-b-(PEG)-PTX and (O-1)-b-(PEG)-HNPs-PTX novel formulations to accumulate and kill pancreatic cancer cells more effectively, compared to the free PTX.

Background: Biological medicines are effective but expensive options for treating patients with chronic and life-threatening diseases unresponsive to conventional small molecule medicines. The importance of biosimilars as an alternative to expensive originator biologics in treatment regimens is increasing year by year. This thesis aimed to examine the prescribing pattern of current marketed biosimilars and understand the potential factors influencing their prescribing in UK. Methods: Examples of generic medicines entry were analysed and compared and contrasted with the entry of biosimilars in both primary and secondary care settings. Web-surveys were conducted with 234 healthcare professionals (HCPs) and 182 service users to investigate their knowledge and attitudes towards biosimilars. Face-to-face semi-structured interviews were then conducted with 26 HCPs and service users to further elucidate interviewee’s perceptions of biosimilars. Results: Findings from the market analyses showed that the penetration of a generic and/or a biosimilar is governed by the cost, the number of products (competitors) and chronicity of use when the delivery devices are the same. When delivery device or another factor influencing prescribers’ perceptions of patient benefit differed, biosimilar uptake was inhibited. The findings from web surveys and interviews showed that both HCPs and service users had a good knowledge and understanding of biosimilars and their importance for cost savings. Concerns about safety and efficacy of biosimilars during the process of switching to biosimilars were expressed by HCPs and services users, although the extent of this concern varied with the clinical discipline. Conclusion: The introduction of biosimilars is associated with considerable cost savings to the NHS. There are subtle differences between specialists’ views on biosimilars and different uptake patterns. Factors influencing prescribing biosimilar medicines were similar but more complicated than generic medicines. Cost is a key driver for the uptake of biosimilars only when patients related factors were justified or equal.

The bone graft procedure is the gold standard therapy for large bone defects and usually involves implanting autologous bone into the defect site. The need to find more patient-friendly alternatives to the bone graft procedure is a driving force behind recent advances in bone tissue engineering. Different synthetic and natural biomaterials are being investigated for their use in the repair of large bone defects. However, there is still a need for scaffold materials that are able to sinter in situ with the capability of delivering growth factors that promote the bone repair process with minimal invasion. In this study, our main aim was to develop a porous, protein-loaded microsphere delivery system with the capability of sintering in situ and with injectable potential. Poly (lactic-co-glycolic acid) (PLGA) was used for this formulation due to its biodegradability, biocompatibility, controllable mechanical properties and good processing capabilities. An optimised procedure for formulating porous and non-porous (protein-loaded) microspheres was established and the microspheres were extensively characterised. There was an inverse relationship between the molecular weight of the PLGA and both the protein release and compressive strength. An intermediate molecular weight (53 kDa) variety of PLGA was chosen for further work based on balancing the need for retaining sufficient compressive strength and a slower protein release profile. The burst release of protein was addressed by investigating various coatings. The combination of chitosan and PLGA to form composite PLGA/chitosan microspheres resulted in the desired reduction in the burst release. The protein-loaded and porous microspheres were combined as a paste and found to sinter at body temperature (37°C) into scaffolds. Whilst previous investigations have focused primarily on a single type of PLGA microsphere (with or without an additional biomaterial), in this study we combined both porous and protein-loaded microspheres into a single delivery system. Furthermore, successful sintering was confirmed when a suspension of the microspheres was injected through a 19 G needle. The biocompatibility and osteogenic potential of the scaffolds were investigated. The composite PLGA/chitosan scaffolds supported the growth of MG-63 osteosarcoma cells and a primary human stem cell line. Furthermore, the scaffolds also supported the osteogenesis of the stem cells, as demonstrated by the presence of the late protein marker of osteogenesis, osteocalcin, and positive von Kossa staining, which is indicative of mineralization. The composite PLGA/chitosan and porous microspheres combine both porosity and the ability to load and sustain the release of protein into one system. Moreover, their ability to sinter at body temperature when injected or applied as a paste demonstrates the dual functionality of the system. This represents a novel approach to delivering protein for tissue regeneration as presently, there has been no report of the combination of PLGA/chitosan microspheres with porous PLGA microspheres as a system that is able to sinter, both post-injection and when packed as a paste, at 37°C. Therefore, the formulation presented in this thesis shows potential for further in vitro and in vivo testing to determine its suitability for bone tissue engineering applications.

Nanoparticles derived from proteins offer a smart material for the design of a new generation of anticancer therapies. In this thesis we describe a variety of novel photodynamic therapy (PDT), drug delivery, and imaging agents that have been combined to produce theranostic systems. The initial research focus was to produce a protein nanocage of Listeria innocua DNA binding protein from starved cells (LiDps) presenting the Gaussia princeps luciferase enzyme (GLuc) on its exterior (GLuc-LiDps) together with the Zn (II)-Protoporphyrin IX (ZnPP) photosensitiser that has been covalently attached to the protein surface. This system operates as a PDT based on Bioluminescence Resonance Energy Transfer (BRET). The cytotoxic effect of GLuc-LiDps-ZnPP nanoparticles in the presence of its substrate (coelenterazine) was tested against two types of breast cancer cell lines: SKBR3, MDA-MB-231, and also the MRC5 non-cancerous cell line, by means of an MTT assay. The results indicated that GLuc-LiDps-ZnPP nanoparticles plus coelenterazine could inhibit the growth and the migration of SKBR3 cells, out of those studied. Downregulation of the Bcl-2 and Mcl-1 anti-apoptotic proteins was also observed after treatment with GLuc-LiDps ZnPP-mediated PDT, suggesting the SKBR3 cells may be undergoing apoptosis. Furthermore, both flow cytometry analysis and confocal microscopy images demonstrated that the GLuc-LiDps-ZnPP appeared to be preferentially internalised in SKBR3 and MDA-MB-231 cell lines without uptake in the MRC5 cell line. Reactive Oxygen Species (ROS) levels were significantly increased in SKBR3 cells compared to MDA-MB-231 cells in the presence of this PDT agent. Similarly, a C-terminal mini Singlet Oxygen Generator (miniSOG) photosensitiser was conjugated to LiDps and human apoferritin (HuAft). Moreover HuAft was fused with the ZHER2:342 Affibody (Afb) targeting peptide to form Afb-HuAft. This 6.7 kDa Afb protein has a high binding affinity for the Human Epidermal Growth Factor receptor (HER2), which is overexpressed on the surface of a number of tumour cells especially breast cancers. Both miniSOG-LiDps and miniSOG-HuAft fusion proteins were successfully expressed and purified and their subunits were self-assembled to form GLuc-LiDps:miniSOG-LiDps and Afb-HuAft:miniSOG-HuAft hybrid chimeric cages. Additionally GLuc was directly fused with miniSOG supplemented with LTVSPWY targeting peptide to produce a novel (GLuc-miniSOG-LTVSPWY) for targeting of HER2 overexpressed cancer cells, potentially applicable for PDT. Further study is required in order to thoroughly characterise the GLuc-miniSOG-LTVSPWY and the hybrid cages as well as determine their respective cytotoxicity. In the second objective, lead sulfide quantum dots (PbS QDs) were utilised as a drug delivery system and an imaging agent. The PbS QDs were capped with a cancer cell targeting agent (mutated Afb (Afb2C)) to form Afb2C-PbS QDs. This construct was further modified by conjugation with ZnPP to produce ZnPP-Afb2C-PbS QDs. The cytotoxic effect of Afb2C-PbS and ZnPP-Afb2C-PbS QDs were studied in vitro using SKBR3 (HER2 positive) and MDA-MB-231 (HER2 negative) cell lines. The results indicate that both types of PbS QDs display anti-proliferative activity against SKBR3 cells through inducement of cancer cell apoptosis and/or necrosis. This was observed from Pre-G1 phase arrest and an increase in cell population in late apoptosis and necrosis. These results may contribute to the development of cancer treatment using of nanoparticles derived from protein.

The pharmaceutical business is driven by innovation and new technologies. In order to improve the overall efficiency, the modern R&D organisations nowadays have integrated problem-solving techniques in their innovation process. This thesis aims to explore and analyse the application of TRIZ technique in the problem-solving process in the medical device sector of the pharmaceutical industry. The findings of the literature review indicate that TRIZ can effectively guide the problem-finding process with a tool kit that can recognise patterns and regularities based on the past solutions in a knowledgebase. The results suggest that such systematic approach is more effective than the conventional methods of trial-and-error. This study conducted a survey amongst the innovative medical device departments of various pharmaceutical companies in the Rhine-Main region in Germany and provided contemporary data on the application of problem-solving tools, especially TRIZ, in those institutions. As a result, the survey data also delivered some possible criteria for technical solutions of medical devices which were subsequently discussed and finalised with a group of experienced experts (expert panel). The next step of the study was organised as a 2x2 experiment. During the experiments, two groups of experienced practitioners were asked to improve the design of two sample medical devices, alternatively using TRIZ and brainstorming. The efficacy of TRIZ application was analysed both in terms of the quality of the technical solutions and that of the group work. The SYMLOG Adjective Rating Form method initiated by Bales was used for the assessment of the group work. The results of the experiment indicate that the impact of the problem-solving tools is influenced by the type of innovation problem. For the analysis of such influences, this research makes a contribution to knowledge by proposing a 2-dimensional framework to capture the problem types. In addition, a TRIZ procedure for the technical innovations of medical devices was developed based on the model of Su et al. Due to the sensitive protection of intellectual property in the pharmaceutical industry, field studies of R&D processes in large pharmaceutical firms are limited in the public literature. This work provides valuable insights into this business sector, especially in respect of application of problem-solving tools and how those tools may potentially improve the outcomes of the R&D activities in the pharmaceutical industry.