Aim: To establish whether Chinese Herbal Medicine (CHM) has any positive effect on human endometrial receptivity to embryo implantation. Design: A prospective interventional clinical study. The measurements used in this study were: serum follicle stimulating hormone (FSH), serum progesterone, endometrial thickness (ET), sub endometrial 3D-PDA ultrasound indices; vascularisation index (VI), flow index (FI), vascularisation flow index (VFI) and pregnancy outcome. Setting: A private London natural female heath practice specialising in treating infertility with Naturopathic and Traditional Chinese Medicine (TCM). The study period was between November 2005 and April 2008. Patient(s): Fifty primary or secondary infertile women who had failed at least two In Vitro Fertilisation (IVF) treatment cycles (the treatment group) with no pathological cause after investigation by infertility specialists and diagnosed as suffering with unexplained infertility. In addition, a reference group of fifty normal healthy volunteer women (the reference group) randomised and double blinded who received either placebo or treatment. Interventions: Both the treatment group and the reference group received CHM for three menstrual cycles. The reference group on a randomised basis received either CHM or placebo. All staff at the clinic and the reference group volunteers were double blinded to who received placebo or treatment until the end of the study. ii Results: The treatment group results indicated that there was a statistically significant positive difference between pre and post treatment for all variables examined (p < 0.001). The reference group results were not as significant but never the less, still indicated that the improvement was statistically significant between the two arms (treatment and placebo) for all measures, with the exception of sub endometrial FI, where there was no significant difference between groups. The number of pregnancies achieved in the treatment group at 6 months post commencement of the last patient was 17 pregnancies, 10 live births, 2 miscarriages and 5 ongoing pregnancies (34% overall pregnancy rate). Conclusions: The study demonstrated that within the treatment group of 50 failed IVF patients CHM did improve all the 6 physiological parameters known to affect endometrial receptivity by a statistically significant amount and provided a 34% pregnancy outcome. The findings of this study show that administration of CHM did have a positive effect on the endometrial receptivity measures used in this study and that the pregnancy rate achieved in the treatment group was encouraging, however without a control arm it is impossible to attribute the pregnancy rate outcome to treatment or reduced stress from the placebo effect of receiving treatment. The study also demonstrated statistically significant improvement in treatment arm of the reference group whilst the placebo arm showed no significant improvement in the same parameters. None of the participants in the study experienced any adverse reactions or side effects from treatment.

There is great interest beginning to be shown in conducting polymers, such as polypyrrole (PPy), for use in the area of drug delivery systems. The main aim of the project was to produce PPy containing antischistosomal compounds and to study their release under electrochemical control. Schistosomiasis is the second most prevalent typical disease in the world, affecting millions of people, the majority of whom are young children. Once affected, parasites live inside the body, causing several terrible symptomatic diseases, such as hepatic fibrosis, ascites and granuloma formation. Antischistosomal compounds investigated for incorporation were niclosamide, niclosamide ethanolamine salt (NES), praziquantel and trichlorfon. All these, apart from NES, were found be to electrochemically inactive in the potential window used to prepare PPy. The conducting polymer grown in the presence of niclosamide was found to have quite different topography to that electrodeposited in its absence, as indicated by light microscopy and scanning electron microscopy (SEM). X-ray photoelectron spectroscopy (XPS) studies, however, did not show any peaks associated with niclosamide. This, together with the very poor solubility of niclosamide in a range of electrolytes (e.g., aqueous Na2SO4 (0.1 mol dm-3); tetrabutylammonium tetrafluoroborate, TBABF4 (0.01 mol dm-3) / acetonitrile; TBABF4 (0.01 mol dm-3) / methanol) suggested that niclosamide was not a suitable candidate for incorporation into this conducting polymer. Nucleation and growth behaviour (chronoamperometry) showed that the presence of praziquantel and trichlorfon in the electrolyte did not,alter PPy formation. Light microscopy and SEM investigations showed that no changes in film topography occured when these two drugs were added to the electrolyte. PPy was found to be electrodeposited at the surface of reticulated vitreous carbon (RVC) electrodes and did not penetrate far into the bulk of the material. Thicker films (7 – 11 μm) and slightly greater penetration was achieved by stepping the potential to +800 mV vs. SCE for 15 min, rather than using potential cycling. Subsequent studies were focused on trichlorfon due to its solubility in aqueous systems and hence greater promise for use in delivery devices to be used in aquatic systems. The presence of trichlorfon in PPy was not evidenced from Fourier-transform infra-red spectroscopy (FTIR) investigations, although this was probably due to the low concentration of the compound, since it was detected by XPS. These studies showed that one trichlorfon molecule was incorporated for every 765 pyrrole units when RVC was used as the substrate, and for every 103 pyrrole units when ITO glass was used. The former substrate has a much larger surface area, however, and so more trichlorfon overall will be incorporated than for PPy on ITO. XPS also showed that one sulphate anion was detected for every 4.6 to 6.4 pyrrole units. Trichlorfon release from PPy was successfully followed by GCMS, where ca. twice as much trichlorfon was released when the PPy/trichlorfon/RVC electrode was held at -300 mV vs. SCE compared to open circuit conditions (no applied potential). At -400 mV vs. SCE, a similar amount of trichlorfon (6.43 μmol) was released after just 60 min compared to that released at -300 mV vs. SCE after 24 h (6.93 μmol). When the potential was too negative (-500 mV vs. SCE), a reduced amount of trichlorfon (4.72 μmol) was released after 60 min compared to the film at -400 mV vs. SCE (6.43 μmol), although it was greater than that released at -300 mV vs. SCE (2.87 μmol) and with no applied potential (1.09 μmol). Thus, trichlorfon was successfully incorporated into PPy and could be released in a controlled fashion by varying the potential. The PPy/trichlorfon/RVC system showed promise for the construction of delivery devices for controlled release of trichlorfon, potentially for use in vivo or in aquatic environments. The last part of the work used computational chemistry techniques to investigate growing conducting polymer chains. Density functional theory (DFT) was used to calculate the unpaired π-electron spin density distribution of oligopyrrole and oligothiophene radical cations using VWN, BLYP and B3LYP functionals. For the oligomers investigated, α,α'-linkages were maintained, which preserve conjugation and hence conductivity. Pendant monomer units, however, joined via α,β'-linkages along the linear chain, were also predicted. The frequency of these pendent groups was dependent on the functional used, with more accurate B3LYP calculations suggesting a higher frequency than those performed using VWN. The time required to perform DFT calculations on long-chain oligomers (ca. 10 monomer units) was still practicable using the high-level B3LYP functional owing to the use of parallel processing. Since the frequency of pendant monomer units along the chain was dependent on the functional used, calculations using the B3LYP functional are recommended over more rapid computations using the VWN functional.

Duchenne muscular dystrophy (DMD) is the second most commonly inherited disorder in man, the phenotype of which displays pathological characteristics of altered skeletal muscle function including, amongst others, abnormal Ca2+ homeostasis and cell signaling. This study used the mdx mouse model of DMD to analyse purinergic responses in dystrophic muscle cells in vitro and skeletal muscles in situ. Initial investigations excluded reduction in ATPe hydrolysing potential from explaining previous observations of heightened nucleotide sensitivities in dystrophic myoblasts. Instead, this study demonstrates for the first time that significant P2X7 receptor abnormalities exist in dystrophic myoblasts and skeletal muscles of the adult mdx mouse; significantly elevated levels of P2X7 receptor mRNA and protein expression were found in primary myoblast cultures, myoblast lines and muscles in situ at 4 months of age and this was extended to analysis of changes in individual P2X7 splice variants. These abnormalities were shown to extend to functional responses in cultured myoblast lines, where heightened P2X7 receptor-specific sensitivity was shown to be associated with significantly higher basal and induced levels of, and altered time course of, extracellular-signal regulated kinase (ERK1/2)phosphorylation in dystrophic myoblasts. ERK activation responses were shown to be inducible by ATP and BzATP stimulation, inhibited by P2X7 antagonists, and unresponsive to ivermectin, thus confirming P2X7 receptor involvement. Similar up-regulation of P2X7 receptor expression coinciding with ERK phosphorylation was demonstrated in mdx muscles in vivo. Additionally, NAD was identified as a mediator of P2X7 responses in dystrophic myoblasts. This study has also employed a mass spectrometry based approach to investigate the immediate downstream effects of P2X7 activation in cultured myoblasts; allowing identification of multiple potential signaling relays for future study that are discussed. The potential for a link between P2X7 receptor and dystrophin expression has been suggested here through the demonstration that abnormalities in P2X7 receptor expression and function are corrected by micro-dystrophin expression in dystrophic myoblasts. In vivo pharmacological P2X7 receptor inhibition using CBBG significantly reduced the number of revertant fibres in dystrophic muscle, indicating a reduction in degenerative/regenerative activity. The data presented in this thesis highlight a novel role for P2X7 receptor signaling in dystrophic myoblasts and muscles in situ; proposing the potential for beneficial therapeutic strategies aimed at manipulating P2X7 signaling responses in vivo, with a view to slowing the progression of what is at present an incurable and invariably fatal disease.

Advances in endoscopic practice are challenging traditional views on how neoplasia should be managed in the gastrointestinal system. Identifying pre-malignant changes in the gut is central to successful early management of most cancers. There has been rapid growth in the equipment and technology available for evaluating potentially neoplastic lesions in the gastrointestinal tract. However, the evidence base for how they should be used is limited. This thesis investigates the use of chromoendoscopy and ‘virtual computed chromoendoscopy’ in the gastrointestinal tract for the identification, assessment and management of early gastrointestinal neoplasia. The first part of the thesis reviews data on the health burden of gastrointestinal neoplasia. It reviews the background research into advanced endoscopic techniques in the colon and oesophagus and outlines the deficiencies in the published literature. Chapters 7-9 describe the development and validation of a new tool (N.A.C.) for invivo histology prediction of colonic polyps <10mm in size using Flexible Spectral Imaging Colour Enhancement (FICE) and indigo carmine (IC) chromoendoscopy without optical magnification. Chapter 7 describes a study to determine the optimum FICE setting for assessment of polyps. Setting 4 came out as the best setting for this purpose. Chapters 8 and 9 describe two studies to define the key surface features associated with neoplastic and non-neoplastic polyps using first a picture based study and then in an in-vivo study. N.A.C. was then used in a prospective cohort study of polyp assessment in a Bowel cancer Screening Population, described in Chapter 10. A prospective study of 232 polyps <10mm in size was performed. FICE and IC significantly improved the accuracy of the in-vivo diagnosis of polyps as compared to WLI endoscopy. In-vivo prediction of polyp histology allowed the endoscopist to set the correct surveillance interval in 83% of cases using WLI alone and in 97% of cases using either FICE or IC based on BSG guidelines. Chapter 11 describes a study which attempts to identify the role of high resolution (HD) endoscopes in lesion characterization. HD endoscopes significantly improved the ability of the endoscopist to make an in vivo diagnosis using FICE compared to standard resolution endoscopes with FICE but had no effect on the WLI or IC predicted diagnosis. Chapter 12 describes the use of acetic acid in a cohort study for the identification of neoplasia within Barrett’s oesophagus to establish the sensitivity and specificity of the technique. Acetic acid chromoendoscopy had a sensitivity of 95.5% and specificity of 80% for the detection of neoplasia. There was a correlation between lesions predicted to be neoplasic by acetic acid and those predicted by histological analysis (r=0.98). There was a significant improvement in the detection of neoplasia using acetic acid compared with white light endoscopy (P=0.001). This method is then refined in a second study by attempting to exploit the aceto-whitening reaction. This is described in Chapter 13. Data from 146 areas of Barrett’s was collected from 121 patients. 84% were male. Mean age 69. In total 72/86 metaplasia, 6/14 LGD, 26/27 HGD, 15/15 IMC and 12/12 areas of invasive cancer were recognised correctly by the endoscopist. A ROC curve was produced for the identification of high risk neoplasia (HGD, IMC and invasive cancer) using the aceto-whitening timings. The area under the curve was 0.93 (95% C.I.0.89-0.97), demonstrating a low probability that a randomly chosen positive case will exceed the value for a randomly chosen negative case. Using a cut off threshold of 142 seconds a sensitivity for neoplasia of 98% (95% C.I. 89-100) and specificity of 84% (95% C.I. 74-91) was achieved. The final chapter of the thesis discusses the clinical impact of the research and how it could be introduced to clinical practice. It reflects on where deficiencies in the published literature still exist where the future research needs are.

Each year, NHS clinical laboratories carry out more than 700 million laboratory tests, of which 50 million are microbiology investigations. Several studies have shown that between 25% and 40% of all tests sent to the laboratory are unnecessary, and up to 46% of ordered microbiology tests are inappropriate. In light of these accounts, the present study was undertaken to evaluate the process of microbiology specimen management in order to assess microbiology test utilisation and the appropriateness of the test ordering processes. The study focussed on respiratory tract specimens using sputum microbiology as a model for the microbiology service inappropriate test utilisation. The overall main aim of this study was to determine the appropriateness of clinical microbiology test utilisation, its clinical relevance and cost-effectiveness, hence recommend better utilisation strategies. A total of 15,941 respiratory tract samples from Barts and The London NHS Trust were randomly selected from the years 2004/05 and analysed retrospectively. Seven hundred microbiology laboratory request forms from patients for whom respiratory tract cultures were requested over a three month period were examined in detail. These requests were derived from 511 sputum specimens, 100 throat swabs, 63 ear swabs and 76 samples from other respiratory tract sites. 641 (91%) of microbiology test requisition forms were completed, provided all requested details by the service users and were therefore considered as appropriate microbiology test requisitions. 660 (94%) of those examined stated the patient’s clinical diagnosis and only in 65 (13%) of these patients was the stated diagnosis as respiratory tract infection. Sixty percent of sputum specimens examined were considered as poor quality. Forty percent of respiratory specimens were reported as culture positive, based on the local hospital criteria of microbiology test reporting. In sputum culture, 39% was reported as culture positive; however, less than 18% were positive with recognised respiratory pathogens, whilst 27% of throat swabs were reported as culture positive, of which 67% had throat pathogens. From the beginning of this study and before, there were no microbiology test comments and interpretation of test results provided with the test result reporting. The test turnaround time of respiratory microbiology results reported within three days in 2004/2005 was only 20%. The total inappropriate respiratory specimens processed locally were 9,575. Extrapolating from our results, this suggests that 2,153,977 nationally were inappropriate in NHS hospitals in 2004/2005. The total cost of inappropriate respiratory microbiology test use was approximately £152,000 in local NHS hospitals. Extrapolating from our results, this suggests that £23,900, 000 nationally was the total cost of inappropriate tests in the NHS hospitals. Following implementation of this study, follow up studies in 2006 and onwards indicated that there has been an improvement in the quality of the microbiology service. The number of good quality sputum specimens was 69% compared to 40% in 2004/2005. While the total microbiology test turnaround time that was reported within three days in 2009/2010 was more than 94%. From mid 2006 onwards, test interpretation comments have been used in all microbiology test result reporting. The total workload of respiratory tract microbiology activity decreased from 18,915/year to 16,651/year over the years 2004/2005 to 2007/2008, which is down nearly 8%. Analysis of the findings showed that the usefulness of culture results was limited by the collection of inappropriate specimens, and lack of clinical information on the microbiology request form. The crucial importance of the role of clinical and nursing staff is stressed if the clinical relevance of sputum culture is to be maximised. The increasing introduction of electronic pathology test requests gives new opportunities to restrict the collection of inappropriate specimens and make substantial savings in resources, both in the ward and the laboratory. This type of study and audit can give invaluable information about the rationale behind testing, and the appropriateness of sampling and transport time. Appropriate measures for corrective actions can be identified.

Introduction: Cancer of the same origin show considerable heterogeneity in sensitivity to chemotherapy both clinically and in vitro, and show rapid adaptation to chemotherapy based on gene expression. This study tested the hypothesis that anticancer drug exposure could render tumour-derived cells more susceptible to second agents, particularly those with specific molecular targets in survival pathways and with the knowledge of cellular pathways, determine new more effective molecularly designed regimens. Materials and Methods: Single agent, combinational and sequential chemosensitivity of a series of cell lines and tumours was assessed using the ATP-based tumour chemosensitivity assay (ATP-TCA). Sensitivity data was correlated with gene expression, measured by quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) in a TaqMan Array following extraction of mRNA from cell samples and standardisation to the housekeeping gene (PBGD). Mutation analysis kits utilising Amplification Refractory Mutation System (ARMS) and scorpion technology were used to establish the presence of activating mutations in EGFR, KRAS, BRAF and PI3K. Results: Heterogeneity in cellular sensitivity to cytotoxic and targeted agents was observed. While gene expression was seen to show some correlation with sensitivity to signalling pathway combination targets, the complexity associated with cellular adaptation prevented the prediction of response to second agent sequential treatment. Discussion: This study has identified potential novel combinations for use in ovarian cancer. This combination of EGFR and PI3K inhibitors has shown greater sensitivity in cell based assays compared with single agent activity and could become the focus of future clinical trials. Successful application of 384 well ATP-based chemosensitivity assays has shown to be a valuable tool for future cell based research. The quantity of viable tumour derived cellular material is diminishing; therefore, methods developed here will continue to provide the means to complete these types of studies in the future.

Rationale: The prevalence of low back pain and associated costs to society are high. Despite this, the number of studies investigating observational data on the quality and costs of care in routine health care services, such as chiropractic, is relatively small in comparison to the clinical trial evidence available on the effectiveness and cost-effectiveness of manual therapies for low back pain. Objective: To document the quality and cost of care in low back pain patients undergoing routine chiropractic care in the United Kingdom. Design: Prospective single cohort multi-centre study. Participants: A sample of 120 chiropractors and 421 patients. Methods: Following the development of a data collection instrument and a pilot study, patients suffering from low back pain were recruited by chiropractic clinics in the United Kingdom. Information was recorded using a patient self-report questionnaire at baseline prior to the initial consultation, and participants were mailed a follow-up questionnaire at three months. Health outcomes, patient experiences of the process and safety of care, and related costs in the intervening three month period were documented. Results: Four hundred and twenty-one patients formed the baseline sample, and 238 (57%) of these returned the follow-up questionnaire at three months. Statistically significant change scores (p = 0.0001) were seen for the health status measures including the Roland-Morris Disability Questionnaire, Bournemouth Questionnaire, EuroQol-5D and bothersomeness scale. One hundred and sixty-eight of 238 (70%) patients reported a clinically significant improvement on the Perceived Global Effect scale, and 73 (31%) of these were considered recovered anytime during the study period using definitions of recovery (i.e. acceptable quality of life, no disability and no pain for a whole month). One hundred and twenty-nine (54%) of patients at follow-up rated chiropractic care for their low back condition as ‘very helpful’. The number of patients rating the process of care (i.e. time and explanations given by chiropractor as well involvement in decisions about care) as ‘very good’ ranged from 157 to 168 (66% to 71% respectively of the patients at follow-up). One hundred and twenty-five (52%) of patients at follow-up reported adverse events of care (i.e. worsening of their back pain, stiffness, soreness and/or general discomfort immediately or shortly after the chiropractic treatment visits); however, only 13 (5%) of these reported that they were unable to carry on with their usual activities and/or work as a result of these events. On average, the total cost of care was £481.83 (95% CI = 333.17 to 639.42) per patient. Lost productivity resulting from time away from work was the most important contributor to these costs (59.6%). The cost of chiropractic visits was the second most important contributor, which accounted for nearly one-third of total costs (32.8%). Other health care usage including general practitioner visits, medical procedures and diagnostic imaging were responsible for a small proportion of total costs ranging from 0.4% to 1.6%. Conclusions: This programme of research is the first prospective study conducted in routine chiropractic practice simultaneously documenting information about health outcomes and patient experiences and costs of care. Patients improved markedly within the first three months of care and expressed high satisfaction with the chiropractic treatment and consultation they received. Chiropractic care was relatively safe, with common yet benign adverse events that had little influence on activities of daily living. Taken overall, patients receiving chiropractic care reported improvement at arguably reasonable cost, suggesting this approach to the health care of patients with low back pain be considered in the wider context of health care delivery in the United Kingdom.

Introduction: Early diagnosis represents the best opportunity for cure of gastro-intestinal cancers; however current gastro-intestinal cancer screening programmes have low test sensitivity and low patient acceptability. It is hoped that a better understanding of carcinogenesis and the development of new biomarkers will provide answers to these clinical problems. Hypothesis and aims: This thesis examines the current understanding of gastro-intestinal carcinogenesis focusing on colorectal and oesophageal cancers. It reviews the circulating gastro-intestinal cancer biomarker literature reporting the strengths, weaknesses and successes of current approaches. The study tests the hypothesis that control patients and patients with colorectal and oesophageal neoplasia have differing plasma levels and fragmentation patterns of cell free nuclear and mitochondrial DNA, and that endoscopic removal of these lesions returns the levels and patterns to normal. We aim to show how new techniques of DNA processing can improve results, and how the application of these results could form part of a diagnostic approach in an at risk population. We compare our results to and including carcinoembryonic antigen levels. Results: Cell free DNA was isolated from 164 patients, including 71 patients with oesophageal neoplasia, 50 patients with colorectal neoplasia, 35 patients without endoscopic abnormality and 8 patients with Barrett’s oesophagus. This is the first report of statistically significant differences in circulating DNA quantities and patterns in patients with early oesophageal and colorectal neoplasia (p≤ 0.005). Logistic regression of the best DNA marker for colorectal neoplasia demonstrated a ROC of 0.888, with a sensitivity of 81.1% and specificity of 75.8%. Logistic regression of the best DNA marker for oesophageal neoplasia demonstrated a ROC of 0.778 with a sensitivity of 81.3% and specificity of 60.5%. Carcinoembryonic antigen performed poorly with a ROC of 0.547 and did not add diagnostically. There were no significant changes in markers from patients resected at endoscopy. Conclusions: These circulating markers in combination with other markers offer the prospect of a simple blood test as a possible screen for colorectal and oesophageal dysplasia and cancers in an at risk population.

Rhodiola rosea, Eleutherococcus senticosus, Epimedium and Panax species have been used in complementary and alternative medicine for thousands of years and are used worldwide for their range of curative effects. In order for any herb preparation to be considered medicinally effective, it must be given at a sufficient dosage level, as well, for therapeutic purposes, the correct plant species is most important. Due to the lack of quality control, many herb-containing products that are made available to consumers on the market today may contain misidentified plant species, counterfeit ingredients, an insufficient quantity of the known active compounds and spiking with marker compounds. Therefore, better regulation and effective control of commercial herbal products are needed. There are three major aims for this research programme: 1) Development of analytical methods for the detection of major biological marker compounds of Rhodiola rosea, Eleutherococcus senticosus, Epimedium and Panax species and establishment of their HPLC profiles. 2) The developed methods from above to be used to evaluate raw materials and commercial products containing the Rhodiola rosea, Eleutherococcus senticosus, Epimedium and Panax species. 3) Establish Rapid Resolution Liquid Chromatography methods for each single herb and analyze multiple chemical constituents simultaneously in formulated products containing Rhodiola rosea, Eleutherococcus senticosus, Epimedium and Panax species. 4) To determine if the proposed developed methods are practical for use, and furthermore, with respect to selected commercial products to clarify that their content matches the claim on their label. Traditionally, single marker compounds are used for the evaluation and determination of herbs. This presents a restriction and consequently many counterfeit products are found on the market. Therefore HPLC profile analysis for the multiple chemical constituent is needed for a more accurate identification of herb species and provides significance for the analysis of formulated products. R. rosea, E. senticosus, Epimedium and Panax species were included in this study as these four herbs are often used in different mixture combinations to make the so called “energy formula”. This includes the simultaneous analysis of up to twenty active ingredients from these four herbs, namely, salidroside, tyrosol, eleutheroside B, E, rosarin, rosavin, rosin, rosiridin, Rg1, Re, epimedin A, B, C, icariin, Rb1, Rc, Rb2 and Rd. A rapid and effective method for the determination of phenylpropanoids, phenylethanol and monoterpenoids from R. rosea, eleutherosides B and E from E. senticosus, flavonol glycosides from Epimedium species and ginsenosides from Panax species in one injection using the reverse-phase Rapid Resolution Liquid Chromatography was successfully developed and used to evaluate the botanical combination. This breakthrough example will lead the industry to optimum quality control methods, especially for formulated botanical products. As well, the results will provide herbalists with more confidence in accepting the so called “standardized botanical extracts”. This developed method can simultaneously analyse Rhodiola, Eleutherococcus, Epimedium and Panax containing products to ensure the correct species.

The central nervous system (CNS) is made up mainly of neurones, the signalling cells, and glia. The main types of glia are astrocytes and oligodendrocytes. Astrocytes are multifunctional cells with vital homeostatic roles, whereas oligodendrocytes are highly specialised to form axonal myelin sheaths, which are essential for rapid neuronal communication. Although glia are involved in every neurodegenerative disease, they have been less well studied as drug targets than neurones. Lithium has been used for decades as a mood stabilising agent and may have major therapeutic potential as a treatment in diverse neurodegenerative diseases. The mode of action of lithium is generally attributed to its potent inhibitory effect on glycogen synthase kinase 3 (GSK3), an enzyme that regulates the Wnt/-catenin pathway. The aim of this study was to examine the effects of lithium and Wnt signalling in glial cells of the adult CNS. First, I developed an organotypic culture model of the adult mouse optic nerve that maintains cellular, axon and myelin integrity in vitro. This model was used to examine the effects of lithium and Wnt on adult glial cells, using confocal microscopy, immunohistochemistry, microarray, western blot and qRT-PCR. A key finding was that lithium acts through the canonical Wnt/β -catenin pathway to stimulate gliogenesis in the adult optic nerve, acting via diverse cell-cycle and cytoskeletal remodelling mechanisms. Lithium profoundly increased oligodendrocytes by downregulating inhibitors of differentiation, such as Wnt, bone morphometric protein (BMP) and inhibitor of differentiation (ID)4. Furthermore, lithium also acted through non-GSK3 targets, including Cxcl1 and leukemia inhibitory factor (LIF), which promote oligodendrocyte differentiation. An important finding was that lithium, and to a lesser extent Wnt, induced the generation of a novel astrocyte morphology, with highly polarised morphology and axon growth promoting phenotype. Notably, I identified a number of novel glial genes in the optic nerve and provide evidence that lithium induces astrocyte polarisation through the inhibition lysyl oxidase and upregulation of the cell-cell adhesion molecule corneodesmosin. In summary, this PhD has demonstrated for the first time that Wnt regulates gliogenesis in the adult CNS and indicates that glial cells are a potential target of lithium in neurodegenerative diseases.