Many chemotherapy drugs act by inducing DNA damage leading to cell death, and the platinum analogue class of anticancer drugs are the most commonly used DNA damaging chemotherapeutic drugs. Despite extensive analysis of platinum-DNA interactions, particularly characterising the individual adducts and their effects on DNA replication, transcription and cell survival, measurement of these adducts in cells with higher sensitivity and precision is necessary. Previous work studying platinum-DNA adduct formation has been performed using DNA damage assays such as immuno-slot-blots to detect whole genome DNA damage, or with combinations of chromatography and mass spectrometry to characterise each adduct individually. The ability to measure platinum-induced DNA damage genome-wide with high resolution in human cells could have profound implications for basic mechanistic research, as well as clinical translational research and treatment stratification, by providing a tool with the potential for predicting clinical response to these agents. The achievement of this PhD was developing an assay to measure platinuminduced DNA damage induction at high resolution, density and precision within the genome of human cells. This was achieved using DNA immunoprecipitation coupled with analysis using DNA microarrays, allowing measurements of platinum-induced DNA damage to be made at high resolution throughout the human genome for the first time. This assay was initially developed to measure cisplatin and oxaliplatin induced DNA damage in the genome of the yeast model organism Saccharomyces cerevisiae and experimental profiles of cisplatin and oxaliplatin-induced DNA damage were validated by demonstrating close correlation with mathematically generated predicted profiles for platinum-induced DNA damage. The assay was then applied and validated to measure cisplatin, oxaliplatin and ultraviolet-induced CPD formation in human fibroblast cells, and again, experimental profiles of cisplatin and oxaliplatin-induced DNA damage and UV-induced CPD formation were shown to correlate well with predicted profiles of DNA damage. Novel comparative analytical approaches for studying microarray data from these genome-wide DNA damage datasets are demonstrated and further validation of the assay is provided by demonstrating the contrast between platinum-induced DNA damage and UV-induced CPD formation genome-wide and in the context of repeat sequences of DNA. Finally, cisplatin and oxaliplatin-induced histone H3 acetylation changes are examined and histone H3 K14 acetylation is demonstrated to be a prominent histone modification following exposure to platinum analogues. Novel analysis is performed to investigate the influence of chromatin on platinum adduct formation, and greater platinum-induced DNA damage is demonstrated in DNA samples treated in vitro compared with DNA samples taken from cells treated in culture. Comparisons were also performed between histone H3 acetylation in yeast cells following exposure to cisplatin or UV irradiation and this comparison revealed very similar patterns of histone acetylation following exposure to these two different genotoxins.

Parkinson’s disease (PD) is the most common neurodegenerative movement disorder, currently affecting 6.3 million people worldwide. Although it is associated, in the longterm, with severe complications (dyskinesias), L-DOPA remains the gold standardtreatment. An alternative approach to the treatment of PD is the replacement of the lost striatal dopaminergic innervation by transplantation of foetal ventral mesencephalon (VM) dopaminergic precursor cells. Opened trials have provided the proof of concept that intrastriatal VM transplant can survive, integrate and in some cases, restore motor functions. Nevertheless, later double blind studies reported inconsistent benefit of the therapy and the development of dyskinesias remaining after withdrawal of L-DOPA medication. The failure of the animal models in predicting these problems raises concern about their reliability. Therefore, the global aim of this PhD work was to identify some of the critical factors that can influence the functional outcome of cell therapy for PD, and on the basis of this, to develop an improved 6-OHDA unilaterally lesioned rat model for transplantation. The first step was to determine the most reliable method to assess dyskinesias in rats. The second part of this thesis was set out to determine the effect that chronic L-DOPA treatment, administered at different time could had on the survival and function of immunologically incompatible foetal VM transplant. The results demonstrated that L-DOPA administered chronically post-grafting increases the host immune response around the xenogeneic transplant. Therefore, the last set of experiments were designed to create a model of mixed donors graft to better reproduce the patient situation, where each transplant required up to 8 donors from unknown immunological background. All of these experiments come together to help to develop a rat model that more accurately represents all aspects of patients undergoing transplantation for PD.

In Chapter 1, I introduce ischaemic stroke, thrombolytic therapy, thrombolysis trials and then discuss the rationale for exclusion criteria in stroke thrombolysis guidelines.In Chapter 2, I describe methods for examining outcomes in patients that are currently recommended for exclusions from receiving alteplase for acute ischaemic stroke. In Chapter 3, I examine Virtual International Stroke Trials Archive (VISTA) data to test whether current European recommendation suggesting exclusion of elderly patients (older than 80 years) from thrombolysis for acute ischaemic stroke is justified. Employing non-randomised controlled comparison of outcomes, I show better outcomes amongst all patients (P < 0.0001; OR, 1.39; 95% CI, 1.26 to 1.54), young patients (P < 0.0001; OR, 1.42; 95% CI, 1.26 to 1.59) and the elderly patients (P = 0.002; OR, 1.34; 95% CI, 1.05 to 1.70). Odds Ratios are consistent across all age deciles > 30 years. Outcomes assessed by National Institutes of Health Scale (NIHSS) score and dichotomised modified Rankin Scale score are consistently similar. In Chapter 4, I compare thrombolysed patients in Safe Implementation of Thrombolysis in Stroke International Stroke Thrombolysis Register (SITS-ISTR) with VISTA non-thrombolysed patients ("comparators" or "controls") and test exactly similar question as in Chapter 3. Distribution of scores on modified Rankin scale are better amongst all thrombolysis patients than controls (odds ratio 1.6, 95% confidence interval 1.5 to 1.7; Cochran-Mantel-Haenszel P<0.001). Association occurs independently amongst patients aged ≤80 (0R 1.6, 95%CI1.5 to 1.7; P<0.001; n=25,789) and in those aged >80 (OR 1.4, 95% CI 1.3 to 1.6; P<0.001; n=3439). Odds ratios are consistent across all 10 year age ranges above 30, and benefit is significant from age 41 to 90; dichotomised outcomes (score on modified Rankin scale 0-1 v 2-6; 0-2 v 3-6; and 6 (death) versus rest) are consistent with the results of ordinal analysis. These findings are consistent with results from VISTA reported in Chapter 3. Age alone should not be a criterion for excluding patients from receiving thrombolytic therapy.In Chapter 5, I employ VISTA data to examine whether patients having diabetes and previous stroke have improved outcomes from use of alteplase in acute ischaemic stroke. Employing a non-randomised controlled comparison, I show that the functional outcomes are better for thrombolysed patients versus nonthrombolysed comparators amongst non-diabetic (P < 0.0001; OR 1.4 [95% CI 1.3-1.6]) and diabetic (P = 0.1; OR 1.3 [95% CI1.05-1.6]) patients. Similarly, outcomes are better for thrombolysed versus nonthrombolysed patients who have not had a prior stroke (P < 0.0001; OR 1.4 [95% CI1.2-1.6]) and those who have (P = 0.02; OR 1.3 [95% CI1.04-1.6]). There is no interaction of diabetes and prior stroke with treatment (P = 0.8). Neurological outcomes (NIHSS) are consistent with functional outcomes (mRS). In Chapter 6, I undertake a non-randomised controlled comparison of SITS-ISTR data with VISTA controls and examine whether patients having diabetes and previous stroke have improved outcomes from use of alteplase in acute ischaemic stroke. I show that adjusted mRS outcomes are better for thrombolysed versus non-thrombolysed comparators amongst patients with diabetes mellitus (OR 1.45[95% CI1.30-1.62], N=5354), previous stroke (OR 1.55[95% CI1.40-1.72], N=4986), or concomitant diabetes mellitus and previous stroke (OR 1.23 [95% CI 0.996-1.52], P=0.05, N=1136), all CMH p<0.0001. These are comparable to outcomes between thrombolysed and non-thrombolysed comparators amongst patients suffering neither diabetes mellitus nor previous stroke: OR=1.53(95%CI 1.42-1.63), p<0.0001, N=19339. There are no interaction between diabetes mellitus and previous stroke with alteplase treatment (t-PA*DM*PS, p=0.5). Present data supports results obtained from the analyses of VISTA data in chapter 5. There is no statistical evidence to recommend exclusion of patients with diabetes and previous stroke from receiving alteplase.In Chapter 7, I examine VISTA data to test whether exclusion of patients having a mild or severe stroke at baseline would be justified. Stratifying baseline stroke severity for quintiles of NIHSS scores, I observe that there are significant associations of use of alteplase with improved outcomes for baseline NIHSS levels from 5 to 24 (p<0.05). This association lose significance for baseline NIHSS categories 1 to 4 (P = 0.8; OR, 1.1; 95% CI, 0.3-4.4; N = 8/161) or ≥ 25 (P = 0.08; OR, 1.1; 95% CI, 0.7-1.9; N = 64/179) when sample sizes are small and confidence interval wide. These findings fail to provide robust evidence to support the use of alteplase in the mild or severe stroke patients, though potential for benefit appears likely.In Chapter 8, I present a meta-analysis of trials that investigated mismatch criteria for patients’ selection to examine whether present evidence supports delayed thrombolysis amongst patients selected according to mismatch criteria. I collate outcome data for patients who were enrolled after 3 hours of stroke onset in thrombolysis trials and had mismatch on pre-treatment imaging. I compare favourable outcome, reperfusion and/or recanalisation, mortality, and symptomatic intracerebral haemorrhage between the thrombolysed and non-thrombolysed groups of patients and the probability of a favourable outcome among patients with successful reperfusion and clinical findings for 3 to 6 versus 6 to 9 hours from post stroke onset. I identify articles describing the DIAS, DIAS II, DEDAS, DEFUSE, and EPITHET trials, giving a total of 502 mismatch patients thrombolysed beyond 3 hours. The combined adjusted odds ratios (a-ORs) for favourable outcomes are greater for patients who had successful reperfusion (a-OR=5.2; 95% CI, 3 to 9; I2=0%). Favourable clinical outcomes are not significantly improved by thrombolysis (a-OR=1.3; 95% CI, 0.8 to 2.0; I2=20.9%). Odds for reperfusion/recanalisation are increased amongst patients who received thrombolytic therapy (a-OR=3.0; 95% CI, 1.6 to 5.8; I2=25.7%). The combined data show a significant increase in mortality after thrombolysis (a-OR=2.4; 95% CI, 1.2 to 4.9; I2=0%), but this is not confirmed when I exclude data from desmoteplase doses that are abandoned in clinical development (a-OR=1.6; 95% CI, 0.7 to 3.7; I2=0%). Symptomatic intracerebral haemorrhage is significantly increased after thrombolysis (a-OR=6.5; 95% CI, 1.2 to 35.4; I2=0%) but not significant after exclusion of abandoned doses of desmoteplase (a-OR=5.4; 95% CI, 0.9 to 31.8; I2=0%). Delayed thrombolysis amongst patients selected according to mismatch imaging is associated with increased reperfusion/recanalisation. Recanalisation/reperfusion is associated with improved outcomes. However, delayed thrombolysis in mismatch patients was not confirmed to improve clinical outcome, although a useful clinical benefit remains possible. Thrombolysis carries a significant risk of symptomatic intracerebral haemorrhage and possibly increased mortality. Criteria to diagnose mismatch are still evolving. Validation of the mismatch selection paradigm is required with a phase III trial. Pending these results, delayed treatment, even according to mismatch selection, cannot be recommended as part of routine care.In Chapter 9, I summarise the findings of my research, discuss its impact on the research community, and discuss weaknesses inherent in registry data and limitation of statistical methods. Then, I elaborate the future directions I may take to further research on the theme of this thesis.

A randomised controlled trial design was chosen to compare a model of community pharmacy based clinical medication review (CMR) with no access to information from patients’ medical records (active) with a model which had access to information from patients’ medical records (control). Four hundred and eighty patients registered with 20 general practitioner (GP) practices received their allocated intervention across 16 pharmacies. There were no statistically significant differences between the groups for the two primary outcome measures used, with 1.7 and 1.9 clinical drug therapy problems (cDTPs) identified at baseline and a cDTP resolution at follow up of 61% and 57% for the active and control groups respectively. There were no significant differences between the groups in the three secondary outcome measures of GP agreement, change in number of repeat medicines and change in utilisation of healthcare services. The cost effectiveness of the two models was compared with the active group found to have a lower cost per cDTP resolved compared with the control group (£46 v £67). Thirty three patients participated in four focus groups with views expressed under the themes of change, empowerment and relationships. Fifteen pharmacists participated in two focus groups and expressed views under the themes of confidence, communication, logistics and empowerment. The hypothesis was accepted such that ‘community pharmacy based CMR of older patients (= 65 years) receiving multiple repeat medicines (= 4) identified and resolved clinical drug therapy problems irrespective of access to information from patients’ medical records’. Both models were well received by patients and practitioners.

Cerebrovascular disease, has an enormous, and increasing, impact on global health. As well as causing clinical stroke, cerebrovascular disease is thought to be a major contributor to cognitive decline and dementia. Socioeconomic status (SES) is associated with risk of stroke. Those in the lowest SES group are estimated to be at twice the risk of stroke compared to those in the highest SES group. Those with low SES may also have a more severe stroke and a poorer outcome. It is imperative that the extent and mechanism of this association is clarified. This thesis aims to determine if the association between SES and stroke is explained by a greater prevalence of traditional vascular risk factors amongst those of low SES. It also explains the link with a novel risk factor, poor oral health. Lastly it addresses the long-term cognitive outcome in older people at risk of vascular disease. A systematic review and meta-analysis was undertaken to establish if vascular risk factors explain the association between SES and stroke incidence / post-stroke mortality. This demonstrated that lower SES was associated with an increased risk of stroke and that a greater burden of vascular risk factors in those with low SES explained about 50% of the additional risk of stroke. However this meta-analysis could not clarify what vascular risk factors are most critical. Low SES was also associated with increased mortality risk in those who have a stroke although study results were heterogeneous and this link was not readily explained by known vascular risk factors. A prospective study of 467 consecutive stroke and transient ischameic attack (TIA) patients from three Scottish hospitals was undertaken with the aim of establishing whether those with low SES carry higher levels of vascular risk factors, have a more severe stroke and have equal access to stroke care services and investigations. Stroke / TIA patients with low SES were younger and more likely to be current smokers but there was no association with other vascular risk factors /co-morbidity. Those who had lower SES had a more severe stroke. The lowest SES group were less likely to have neuroimaging or an electrocardiogram although differences were not significant on multivariate analysis. There was however equal access to stroke unit care. A secondary analysis of a prospective cohort study of 412 stroke patients was conducted. The aim was to explore oral health after acute stroke and assess if poor oral health explains the association between SES and stroke. Dry mouth amongst acute stroke patients was very common, however there was no association between oral health and low SES. There was an association of dry mouth with pre-stroke disability and Urinary Tract Infection. There was also a link with oral Candida glabrata colonisation, although the clinical relevance of this is uncertain. In the acute phase after stroke there was no convincing association of dry mouth with dysphagia or pneumonia. Therefore there was no association between SES and poor oral health as measured in this study but oral health may still be part of the explanation of the association between SES and acute stroke and this needs further investigation. Vascular disease is an important contributor to cognitive decline and dementia. Low SES may be associated with an increased risk of cognitive decline in later life and vascular disease may be a mediating factor. More effective prevention of vascular disease may slow cognitive decline and prevent dementia in later life, particularly in low SES groups. Lipid lowering with statins might be effective in preventing dementia but so far evidence from randomised control trials does not show benefit from statins in preventing cognitive decline and dementia. However the duration of follow-up in these trials was short and there may be benefit in the long-term. My aim was therefore to establish if long-term follow-up of the Prospective Study of Pravastatin in the Elderly at Risk (PROSPER) study was feasible. I found that it was feasible to follow-up 300 elderly survivors from the Scottish arm of the PROSPER study and the methods could be extended to the whole group. As expected nearly half of the PROSPER participants were dead. Additionally a large proportion of traceable participants had significant cognitive impairment. Smoking cessation, control of blood pressure and management of other vascular risk factors should be made a priority in areas of low SES. Additionally further research is needed to fully clarify the association between SES and stroke incidence. Avenues for exploration might include the possibilities of poorer access to effective stroke care, reduced uptake of care and poorer oral health in lower SES groups. In addition public health campaigns regarding smoking cessation should be directed at lower SES groups. I have shown that a large scale follow-up of the PROSPER participants is feasible and may determine new and novel risk factors for dementia and assess the long-term effect of a period of treatment with pravastatin.

Acyclovir (ACV) is an acyclic guanine nucleoside analogue used in the treatment of herpes simplex virus (HSV) and varicella zoster virus (VZV) infections. The human herpes virus-encoded thymidine kinase (HHV-TK) phosphorylates ACV generating ACV monophosphate, which is then converted to the active form ACV triphosphate by cellular kinases. The ProTide approach, causing the direct release of the monophosphate form into the cell, allows bypass of the first phosphorylation step of nucleoside analogues. In previous studies acyclovir ProTides were found to be active in vitro against human immunodeficiency virus (HIV), demonstrating a successful release of ACV monophosphate into the cell. In this work, an extensive study of structure-activity relationship was carried out varying the masking groups of the ACV ProTide. Subsequently, several substituents were considered on specific position of the guanine base and side chain of ACV resulting in the synthesis of the aryl phosphoramidate derivatives of ganciclovir, penciclovir, 6-O-alkyl acyclovir, 8-bromoacyclovir, and 8- methylacyclovir. These derivatives were evaluated in vitro for their antiviral activity against HSV, VZV, human cytomegalovirus (HCMV), and HIV. Enzymatic and molecular modeling studies were carried out in order to investigate the bioactivaton of the phosphoramidate derivatives synthesised in this work and correlate these results with their biological activity. Finally, the ProTide approach was also applied to cidofovir resulting in the formation of the phosphonoamidate derivative of cyclic cidofovir. The in vitro antiviral activity of this compound against herpes viruses and poxviruses is reported, as well as investigation of the mechanism of activation, using enzymatic and molecular modeling studies.

Bipolar disorder is a severe and enduring mental health condition. Although early identification is associated with better outcomes, research has shown that many people wait over a decade after first experiencing affective symptoms before a correct diagnosis is made. A misdiagnosis of unipolar depression often leads to the inappropriate prescription of potentially mood-destabilising antidepressant medications. The aim of this study was to consider service user and carer experiences and their often overlooked views on diagnostic issues. Participants were recruited via the charity Bipolar UK. Initially a focus group discussed the diagnostic process and relevant issues, then the transcript was analysed thematically and used to develop a detailed survey questionnaire. The survey was piloted before being launched online and completed by 262 service users and 65 carers. Survey respondents reported an average delay of over eleven years between first seeking help for bipolar symptoms and being correctly diagnosed. Almost 70% reported a previous misdiagnosis of depression, and over half had experienced mania/hypomania whilst taking antidepressant medication. Nearly a third consulted a doctor over 20 times with bipolar symptoms before receiving help for bipolar disorder. Participants considered that improvements are needed in primary care mental health assessment and that GPs need more training. Many reported that their condition had led them to be discriminated against and agreed that society’s lack of understanding was a major problem. Carers indicated similar opinions and emphasised the potential benefits of including carers in the assessment process. This study suggests that the identification of bipolar disorder is often poor and that assessment of the condition in primary care needs to be improved. Service users offered suggestions for improvements, including recognition of early indicators, appropriate assessment questions and better training which could be adopted by service providers. Limitations of the study and implications for future research were discussed.

The search for new anti-HCV therapeutics continues, as current standard of care based on pegINF and ribavirin is limited and can cause significant side effects. Several families of modified nucleoside are known to inhibit HCV RNA dependent RNA polymerase (RdRp). The 2’-C-!-methylguanosine (2’CMeG) has been identified as one of the most potent RdRp inhibitors (IC50 = 0.26 μM as NTP). Despite its very good activity at the triphosphate level its poor cell uptake and presumably poor phosphorylation lead to significant decrease in active in HCV replicon assays (EC50 = 3.5μM). The ProTide approach was applied to 2’-Cmethylguanosine in order to by-pass the first phosphorylation limiting step and enhance cellular uptake. Modified nucleoside phosphoramidates with improved efficacy and selectivity may become a future of HCV therapies. In the present work, the pronucleotide approach based on aryloxyphosphoramidate, phosphorodiamidate or oxazaphosphorine cyclic prodrugs has been applied to 2’-C-!-methylguanosine and related nucleoside analogues bearing modifications in C6-, C8- and C2- of the purine base. These modifications were mainly introduced to increase lipophilicity of 2’CMeG and consequently enhance cellular uptake and to deliver the 2’-C-methylguanosine 5’-monophosphate intracellularly. In general, most of the newly synthesised compounds exhibited excellent potency in HCV replicon assay. The most potent compounds were up to 1000-fold more active than the corresponding parent nucleoside. Base modification combined with the ProTide approach provide compounds that possess not only excellent antiviral activity but also good cell permeability. Extensive in vitro and in vivo studies lead to the selection of INX-08189 that has now progressed into human clinical trials for HCV, being currently in phase IIa clinical development.

In the treatment of pre-menopausal women with oestrogen positive (ER+) breast cancer, tamoxifen represents a first line of adjuvant treatment with demonstrable benefits. Despite this, resistance is frequently acquired to tamoxifen with an associated poor prognosis. Breast cancer cell models have revealed the importance of growth factor signalling networks in sustaining growth of endocrine-resistant cancers and, more recently, their ability to promote a highly migratory and invasive phenotype, together with the expression of genes with pro-angiogenic ontology. The potential of endocrine-resistant cells to elicit angiogenic responses, however, remains unknown. Real-time PCR was used to validate results from preliminary Affymetrix-based gene profiling of pro-angiogenic gene expression in endocrine-sensitive MCF7wt cells and their endocrine resistant counterparts. The expression of pro-angiogenic factors in conditioned media (CM) from these cells was assessed by ELISA. The proliferative and migratory effects of conditioned media on vascular endothelial cells (HUVEC and HECV cells), was determined by MTS cell proliferation assay, wound closure assays and Matrigel tubule formation assays. Changes in endothelial cell migration following co-culture with endocrine-resistant cells were examined using Boyden-chamber chemotaxis assays. Growth factor signalling and migration pathway activation in endothelial cells in response to CM was determined by Western blotting. TamR cells were found to express high levels of IL-8 and VEGF at an mRNA level compared with expression in MCF7wt cells. High levels of VEGF protein were also confirmed in the conditioned media from TamR cells versus their endocrine-sensitive counterparts. TamR conditioned media promoted in vivo and ex vivo endothelial cell proliferation, as well as in vitro endothelial cell migration and the formation of tubules to a greater extent than that seen in MCF7wt CM treated cells. TamR conditioned media was found to stimulate VEGFR2 phosphorylation and downstream activation of MAPK and Akt in endothelial cells compared to MCF7wt CM. Pharmacological inhibition of VEGFR2 activity in endothelial cells suppressed TamR-induced endothelial cell proliferation and VEGFR phosphorylation. Further pharmacological manipulation of Src kinase in TamR cells revealed a Src kinase dependent mechanism of VEGF production in these cells. In addition, in vivo examination of TamR xenografts illustrated higher presence of endothelial cells in these tissues than in MCF7wt xenografts. These data suggest acquired tamoxifen resistance is accompanied by development of a Src kinase-dependant pro-angiogenic phenotype which, if recapitulated in vivo, may promote tumour progression. Therapeutic targeting of Src signalling may prove beneficial in such cases.

In this thesis, the behavioral, electrographic and neurobiological effects of a period of kainic acid-induced status epilepticus (SE) on the C57BL/6J inbred mouse strain are characterised. The severity of epileptic behaviour was scored, used immunohistochemistry to investigate the anatomical distribution of c-Fos expression in the hippocampal formation following SE and recorded EEG during and after SE using an implantable, wireless telemetry device. Further to assessing the severity of SE, changes subsequent to seizures related to the emergence of chronic epilepsy were investigated, including reactive gliosis and synaptogenesis and epileptiform discharges in the EEG trace. I investigated the potential of a range of pharmacological agents for modulating the severity of induced seizures and disease progression. These included drugs targetting the NR2B subunit of the NMDA receptor (RO 25-6981), neuronal nitric oxide synthase (L-NPA), The post synaptic density protein 95 (Tat-NR2B9a) and inducible nitric oxide synthase (1400W). L-NPA, when administered prior to the induction of SE was found to profoundly suppress the emergence of epileptiform activity, including behavioural, electrographic and neurobiological indicators. Further, L-NPA’s modulation of the precipitating event lead to a decrease in neurobiological changes associated with epileptogenesis, such as reactive gliosis in the CA3 region of the hippocampus and 5 elevated synaptogenesis in th molecular layer of the hippocampus. This correlated with a marked decrease in epileptiform discharges in the EEG trace. A novel method of kainic acid administration was trialed, involving multiple small doses of the drug, titrated by the severity of behaviour. This method led to a decrease in mortality and an increase in the severity and inter-individual uniformity of SE, assessed by the analysis of behaviour, EEG and c-Fos expression in the hippocampus. Furthermore, this method induced neurobiological changes associated with epileptogenesis 3 days following SE and was associated with an increased frequency of epileptiform discharges for 7 days post SE.