Tissue microarrays of prostate tumours were stained with an antibody to the IGF IR. IGF IR expression was significantly associated with Gleason grade with IGF IR expression higher in Gleason grade 3 relative to normal/benign prostatic hyperplasia (BPH) tissue and lower in Gleason grade 5 relative to Gleason grade 3 tissue. There was no significant difference in plasma IGF I levels between BPH and prostate cancer patients (Gleason score 5 or Gleason score 7). However, there were significantly higher plasma levels of the IGFBP 4 protease, PAPP-A, in Gleason score 5 patients relative to BPH patients.

PC-3M-luc2 cells are a human androgen independent cell line lacking the androgen receptor. The PC-3M-luc2 cells stably express luciferase allowing them to be imaged in v iv o using bioluminescent imaging. Expression of the IGF pathway components was assessed in these cells to ensure the IGF pathway was active. Western blot analysis showed that the PC-3M-luc2 cells expressed the IGF IR which was activated by IGF I. IGFBP 3, IGFBP 4 and PAPP-A were also expressed. PC-3M-luc2 cells were shown to proliferate in response to IGF (E3R) (an analogue of IGF I that cannot be bound by IGFBPs) by bromodeoxyuridine (BrdU) assay. However, the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay showed PC-3M-luc2 cells proliferated in response to IGF I. IGF (E3R) also significantly increased vascular endothelial growth factor (VEGFies) expression in PC-3M-luc2 cells.

We assessed a PAPP-A resistant mouse IGFBP 4 (dBP4) as a strategy to neutralise the IGF I ligand. dBP4 was developed by mutation of the PAPP-A cleavage site from 119-KHMAKVRDIRSKMK-133 to 119-AAMAAVADASAMA-133 and cloned into pTriEx4 Neoplasmid with C-terminal His-tag for expression in human embryonic kidney cells. Purified dBP4 was resistant to PAPP-A cleavage and retained IGF I binding capacity. In v itro , dBP4 significantly inhibited tubule formation by endothelial cells relative to controls or IGF I treated cells suggesting that dBP4 possesses anti-angiogenic properties.

A Tet inducible expression system was used in v iv o to test the effect of dBP4 on tumour growth and metastasis in subcutaneous, orthotopic and experimental metastasis models of prostate cancer. The inducible Tet-On system allowed dBP4 expression to be induced by doxycycline. Following implantation of PC-3M-luc2 dBP4 cells into non-obese diabetic, severe combined immunodeficiency, gamma 2 receptor null (NSG) mice, tumours and metastasis were monitored by bioluminescent imaging and/or caliper measurements. In the subcutaneous tumour model, dBP4-expressing subcutaneous tumours were significantly smaller than controls and in the orthotopic model, bioluminescence was reduced relative to controls. Mice bearing dBP4-expressing subcutaneous tumours survived significantly longer than controls. There was no survival difference between mice bearing dBP4-expressing prostate tumours and controls. IGF I expression within the dBP4-expressing subcutaneous and prostate tumours was decreased relative to controls. Western blot analysis of tumour lysates showed that dBP4 inhibited IGF IR activation in both the subcutaneous and prostate tumours, which may be a consequence of reduced IGF I within the dBP4-expressing tumours. However, activated Akt was only decreased in the subcutaneous tumours relative to controls. As tumours require vasculature in order to supply oxygen and nutrients essential for tumour growth, expression of VEGF was assessed in the subcutaneous tumours. Control tumours expressed VEGF164/165, which was not expressed in the dBP4-expressing subcutaneous tumours.

dBP4 expression inhibited tumour growth in both subcutaneous and orthotopic-prostate cancer models. As dBP4 expression inhibited endothelial cell tubule formation and abolished VEGF_164/165 expression within PC-3M-luc2 tumours growing subcutaneously, this

suggests that dBP4 has either anti-angiogenic or anti-proliferative properties and may

therefore have value as a therapeutic for prostate cancer.

In Study 1, young people with 22ql 1 deletion syndrome were found to have significant differences in both white matter microstructure and volume. Additionally, there was preliminary evidence that within 22ql 1 deletion syndrome, some regional differences in fractional anisotropy were associated with allelic variation in COMT and with schizotypy. In Study 2, while significant grey and white matter volume deficits were found in temporal lobe epilepsy with comorbid psychosis, these abnormalities encompassed not only the medial temporal lobe structures but also extended to lateral temporal and extratemporal regions whereby some of the deficits also overlapped with those found in schizophrenia.

In Study 3, reduced fractional anisotropy was found in antisocial personality disorder and psychopathy in tracts of interhemispheric, posterior brain and frontal lobe networks. Additionally, fractional anisotropy deficits in the frontal lobe demonstrated a significant negative correlation with psychopathy measures. Finally, in Study 4, adults with Asperger syndrome were specifically recruited and found to not only demonstrate impairments in white matter microstructural integrity in regions relevant to social skills and behaviour but also in more widespread white matter networks.

Platelet activation was significantly diminished in the presence of either reducing GSH/GSSG or Cys/CySS redox potentials. Moreover, this response was most notable when platelets were activated with collagen only. The redox modulated effects were shown to be specific to the integrin ɑ₂β₁, through a mechanism involving disulphide bond reduction. An analysis of the platelet integrin β subunits revealed the presence of the greatest number of potentially reactive cysteine residues within the Pi subunit when compared to the other platelet integrin beta subunit β₃.

Furthermore, platelet proteins were S-glutathionylated in a reducing extracellular redox environment. The findings of this thesis reveal for the first time how exquisitely sensitive collagen-induced platelet activation is to alterations in the external redox environment. This, in fact, is potentially a novel demonstration of reductive stress regulating platelet reactivity and provides a novel avenue for therapeutic intervention using physiological redox reagents or their precursors.

The aims of the study were to examine: 1) the mental health outcomes and social and occupational functioning of a cohort of young people (now aged 19-23 years), 2) the association between life stresses such as poor family functioning, parental separation / divorce and peer relations, substance use and anti-social behaviour and mental health outcomes in young adulthood.

Method: Two hundred and twelve young people, who participated in the original Challenging Times Study, were invited to participate in an assessment interview. Mental health outcomes and social and occupational functioning were assessed using the Structured Clinical Interview for DSM-IV psychiatric diagnoses (SCID I), the Achenbach Adult Self Report and Achenbach Adult Behaviour Checklist (parental questionnaire).The Me Master Family Assessment Device was used as a measure of family functioning whilst the Global Assessment of Functioning (GAF) scale measured quality of life and general functioning. The Stressful Life Events Schedule for Children and Adolescents a semi-structured interview was used to identify factors associated with mental ill-health.

Results: Using a weighted population prevalence analysis, 24.3% (22.4 - 28.9) of the young people met the criteria for a psychiatric disorder at follow-up, including 5.4% (4.1 - 7.6) with a mood disorder, 11.2% (9.5 -14.3) with an anxiety disorder, 8.2% (6.7 -11.0) with a substance use disorder and 7.7% (6.2 -10.4) with alcohol disorders. Logistic regression revealed that an adolescent diagnosis of being “at risk” of a psychiatric disorder at the Challenging Times original study was not predictive of poor mental health outcomes at follow-up. Specific stressful life events were identified as factors associated with poor mental health outcomes.

Conclusion: This study offers an understanding of the prevalence rates and psychosocial factors associated with psychiatric disorders in young adulthood in an Irish context. It emphasises the need for youth mental health Intervention and prevention programmes that equip young people with the necessary coping skills to deal with life’s many challenges.

A mixed-methods design was adopted. A prospective cohort study was initially conducted with 145 participants with chest pain who attended exercise stress testing and had normal test results. At one-year follow-up, 69% reported continued pain. In addition, nearly half of participants had returned to their general practitioner and one in ten had attended the emergency department for the investigation of chest pain. In logistic regression analyses, the variables heartburn, pain precipitated by movement, cardiac anxiety, illness perceptions, and lack of communication about test results were predictive of persistent chest pain. When participants with continued chest pain were categorised into persistent healthcare users and non-persistent healthcare users, these variables were predominantly associated with participants with persistent health service use for chest pain. In addition, a number of psychological variables including anxiety and depression distinguished the persistent service users. Employment appeared to be a protective factor against persistent pain and related service use.

A small sample of participants from this cohort was interviewed in a qualitative study informed by the principles of Interpretative Phenomenological Analysis (IPA). Analysis revealed three predominant themes: 1) the disempowerment of normal test results; 2) limbo - the inner struggle of negating and relating to potential causes; and 3) the inadequacy of healthcare to validate and care for symptoms. The dynamic, complex process of interpreting symptoms and deciding whether to seek healthcare was illuminated.

The results indicate that interventions targeting the assessment of a potential gastro-oesophageal or musculoskeletal cause and the reduction of cardiac anxiety are likely to improve outcomes in these patients. Improved communication with patients is also indicated. Psychological factors appear to drive persistent service use and interventions targeting these are likely to reduce medical costs. Patients with NCCP are not a homogenous patient group and an individualised, stepped-care approach to management appears to be warranted.

A mixed-methods design was adopted. A prospective cohort study was initially conducted with 145 participants with chest pain who attended exercise stress testing and had normal test results. At one-year follow-up, 69% reported continued pain. In addition, nearly half of participants had returned to their general practitioner and one in ten had attended the emergency department for the investigation of chest pain. In logistic regression analyses, the variables heartburn, pain precipitated by movement, cardiac anxiety, illness perceptions, and lack of communication about test results were predictive of persistent chest pain. When participants with continued chest pain were categorised into persistent healthcare users and non-persistent healthcare users, these variables were predominantly associated with participants with persistent health service use for chest pain. In addition, a number of psychological variables including anxiety and depression distinguished the persistent service users. Employment appeared to be a protective factor against persistent pain and related service use.

A small sample of participants from this cohort was interviewed in a qualitative study informed by the principles of Interpretative Phenomenological Analysis (IPA). Analysis revealed three predominant themes: 1) the disempowerment of normal test results; 2) limbo - the inner struggle of negating and relating to potential causes; and 3) the inadequacy of healthcare to validate and care for symptoms. The dynamic, complex process of interpreting symptoms and deciding whether to seek healthcare was illuminated.

The results indicate that interventions targeting the assessment of a potential gastro-oesophageal or musculoskeletal cause and the reduction of cardiac anxiety are likely to improve outcomes in these patients. Improved communication with patients is also indicated. Psychological factors appear to drive persistent service use and interventions targeting these are likely to reduce medical costs. Patients with NCCP are not a homogenous patient group and an individualised, stepped-care approach to management appears to be warranted.

Experiments in this thesis were aimed at providing insight into a new localization of Foxo3a within mitochondria. Presently, it has been shown that Foxo3a was localized to mitochondria in several organs in mice, and that Foxo3a binds to the regulatory region of the mitochondrial DNA. The localization of Foxo3a within hippocampal mitochondria of mice was altered during experimentally induced status epilepticus (SE). The mitochondrial level of Foxo3a dropped dramatically after SE. Notably, a drop in mtDNA transcription was observed concomitant with the drop of Foxo3a levels in mouse hippocampal mitochondria. Experiments in this thesis also demonstrate that transcription of mitochondrial DNA is enhanced by the overexpression of Foxo3a in a hippocampal cell line HT-22. Finally, the overexpression of a mitochondrially targeted Foxo3a fusion protein led to a significant protection of the HT-22 cells against cell death induceexposure to glutamate or epoxomicin, but not staurosporine. These data suggest a novel physiological role for Foxo3a in mitochondria, protecting the cell from harmful insults caused by stressors such as oxidative stress and possibly by regulation of the mtDNA.

We observed a delayed calpain activation that occurred downstream of mitochondrial engagement and directly preceded neuronal death. In contrast, we could not detect significant calpain activity during excitotoxic necrosis or in neurons that were tolerant to excitotoxic injuiy. Oxygen/glucose deprivationinduced injury in organotypic hippocampal slice cultures confirmed that calpains were specifically activated during .ax-dependent apoptosis and in this setting function as downstream cell death executioners. Lastly, box gene deletion prevented NMDA-induced alterations to Ay/m, hyperpolarisation and depolarisation were notably diminished, and affected neuronal calcium handling, resulting in increased neuronal survival.

Targeting anti-tubercular therapeutics to alveolar macrophages using microparticle technology aims to reduce systemic toxicity, increase local concentrations of therapeutics and potentially reducing the frequency of dosing requirements. Localising therapy increases local concentration levels of anti-tubercular therapeutics, however, if these therapeutics are encapsulated into a particulate delivery system they can have longer retention time in the lungs also. There has been extensive research into inhaled TB therapies to target anti-MTb drugs directly to the lungs. The work presented herein sought to improve alveolar macrophage targeting by preparing microparticles with optimal targeting properties containing anti-tubercular molecules. The use of novel excipients and microparticles in respiratory drug delivery is limited by the understanding of how these molecules interact with biological systems. The use of high content imaging platforms both in vitro and in vivoemployed within this thesis will serve as a framework for development and testing of other respiratory delivery systems. The microparticles used in this study are based on poly (lactide-co-glycolic) acid(PLGA), a biodegradable polymer broken down through non-enzymatic hydrolysis into lactic and glycolic acid monomers.

This polymer is currently FDA approved for non-pulmonary use and by developing a robust understanding of particle - cell interaction, it might bolster the argument for approval of its use in the lungs for specific clinical indications such as TB.

This project has developed bioengineered microparticle carrier systems to efficiently target the alveolar macrophage, the niche environment of the pathogen. These microparticle delivery systems are composed of a biodegradable polymer (PLGA), which encapsulates a drug with the goal of achieving optimal cellular concentrations of the therapeutic within the cell. The work has demonstrated that 2.1pm sized microparticles are optimal for targeting alveolar macrophages in vitro without affecting cellular viability. The use of coatings such as gelatin (Type A) significantly enhanced microparticle uptake by THP-1 cells and alveolar macrohpages obtained from bronchoalveolar lavage of human volunteers.

The compatibility of these microparticles with a range of anti-tubercular drugs (PAS, rifampicin and capreomycin) has also been demonstrated. Drug loaded microparticles exhibited a controlled release profile over 21 days and were shown to be effective at impairing Mycobacterium tuberculosis (MTb)growth in an in vitro cell model.

Several obstacles must be overcome before a successful inhalable therapy can be developed. In order for an inhalable anti-tubercular formulation to be successful it must be capable of depositing in the alveolar region of the lungs. These obstacles include patient physiology, device selection, powder flowability and aerosol properties. All these parameters are inter-linked. The device employed (Spinhaler) is a single dose drypowder inhaler device that is simple to use, cheap and a high respirable fraction was achieved with this device. Optimisation of the lyophilisation process enhanced aerosol performance while the addition of mannitol (5% w/w) to coated microparticles was required for optimal aerosol performance. However, a caveat to this is that it was achieved at a high flow-rate, which may not be representative of patients with severely impaired lung function. Newer dry-powder inhaler devices are capable of achieving high respirable doses at low flow-rates and may be used to further develop the technology described in this thesis.

Simulating the phagocyte is an exciting concept in the design of novel anti-tubercular therapies and in addition to drug targeting to alveolar macrophages, the microparticle delivery systems showed potential to modulate innate immune responses to MTb infection. Microparticles coated with gelatin (Type A) produced the highest level of NFkB induction and autophagasome formation. This resulted in a significant reduction of MTb viability when infected cells were treated with drug-free coated microparticles. Uncoated microparticles gave a non-significant decrease in MTb viability.

Combining the immune activating properties of these microparticles with the chemotherapeutic effect of anti-tubercular drugs provides a new formulation for old drugs that could improve sputum conversion times and* by localising this therapy to the lung, non-specific systemic side-effects could be avoided.

Finally, a novel coating of Interferon-gamma (IFN-y) on these drug loaded microparticles was designed to achieve maximal immune, stimulation with the combinatorial effect of the chemotherapeutic (rifampicin). This formulation showed promise in vitro but the small pilot study suggested potential immunopathology caused by the IFN-y coated microparticles. Optimisation of the dosing regimen may be required for further in vivo development. The in vitro work establishes this combinatorial potential of an immune modulating agent on the microparticle surface combined with a chemotherapeutic inside the particle, particularly with reference to resistance strains of MTb infection. The establishment of an in vivo imaging system to measure mycobacterial growth / killing in real-time herein will also expedite development of further novel inhalable anti-tubercular formulations.

Proteasome inhibitors have also been used to re-sensitise resistant cancer cells to TRAIL treatment. Since no kinetic data from single cells are available yet, it is not known whether, when and where the intracellular signalling kinetics of TRAIL-induced apoptosis are affected by proteasome inhibition. We therefore quantified the signalling kinetics of TRAIL-induced apoptosis in response to proteasome inhibition in single cells. Depending on the TRAIL concentration we found two modulation sites, which interfered with TRAIL signalling kinetics upon proteasome inhibition: caspase-8 activation at low doses and the threshold for mitochondria1 permeabilisation at high doses. Our results suggest that upregulation of cFLIP and Mcl-1 by impaired protein degradation were responsible for delayed caspase-8 activation and prolonged caspase-8 activation before MOMP, respectively. Our findings therefore indicate that the synergy between TRAIL and proteasome inhibition is probably based on the stabilisation of active caspases rather than enhanced DISC formation, as is often proposed in the literature.

To identify novel SRC-1 target genes luminal B LY2 breast cancer cells were used to perform the first ChlP-sequencing experiment of SRC-1. Affymetrix whole genome expression arrays were used to analyse gene expression in LY2 cells treated with SRC-1 siRNA in comparison with those treated scrambled siRNA to complement the ChlP-seq experiment. SRC-1 peaks within the genome were focused near transcription starts sites and often in close proximity to an estrogen response element. Three new putative SRC-1 target genes were selected for further validation.

ADAM22 is a transmembrane disintegrin protein that has been shown to mediate cell migration but has yet to be implicated in tumourigenesis. Knockout, overexpression and ChlP studies confirm ADAM22 as an SRC-1 target gene. Expression of ADAM22 was identified in 49% of patients with breast cancer and its expression was associated with an almost two fold increase in rate of disease relapse. Two other genes, namely BCAS3 and CUXl were identified and validated as SRC-1 target genes and both of these have a known role in breast tumour progression. Elevated levels of SlOOB were detectable in the blood of 10% of patients with breast cancer. Elevated serum SlOOB at time of diagnosis is an independent predictor of disease progression in breast cancer.

An unbiased, genome wide approach has identified a variety of novel, direct SRC-1 target genes. A series of molecular and translational studies have validated SlOOB as a clinically important and detectable target of SRC-1 while ADAM22, as transmembrane protein may offer the first therapeutic target to disrupt the central role of SRC-1 in mediating breast cancer metastasis.

This study was performed to gain insights into the mechanism of angiogenininduced neuroprotection via surrounding glial cells focusing on astrocytes. The vesicular internalisation of recombinant human angiogenin by primary astrocytes was investigated in culture using pharmacological inhibitors, cell transfection and immunocytochernistry. This revealed that angiogenin endocytosis by astrocytes is clathrin-dependent and involves dynamin for vesicle scission. Vesicle budding and trafficking do not require a functional microtubule network. A fraction of the internalised angiogenin is targeted for lysosomal degradation, while the majority remains in uncharacterised sorting endosomes. The receptor for angiogenin on astrocytes was identified to be the heparansulfate proteoglycan syndecan 4 by colocalisation studies and protein knock down.

To further elucidate the role of angiogenin in paracrine neuroprotection, modification of the astrocyte secretome was investigated in response to angiogenin treatment by quantitative mass spectrometry. Metabolic protein labelling by Stable Isotope Labelling with Amino acids in Culture (SILAC) was optimised for primary astrocytes and proteins were identified by Fourier transform tandem mass spectrometry (nano-LC-FT-MSIMS). This screen identified over 1,500 proteins in the supernatant of primary astrocytes, many of which are known to exhibit extracellular location like components of the Extracellular Matrix (ECM), cytokines, growth factors and growth factor binding proteins. However, proteins with established intracellular function like splicing factors and ribosomal proteins were detected, too. Quantification using the MaxQuant software showed significant regulation of over 100 proteins in response to angiogenin treatment. The most strongly regulated proteins are involved in modifying the ECM or contribute to immune responses and might be responsible for the neuroprotective effects of angiogenin. These data also suggest the involvement of surrounding immune and endothelial cells in the biological activity of angiogenin. Additionally, local transfer of factors involved in protein translation from astrocytes to surrounding cells may be affected by angiogenin.

In conclusion, this study has shed new light on the role of angiogenin in the complex cellular interplay in the Central Nervous System (CNS) focusing on astrocytes and how they interact with neighbouring cells. Further studies will be necessary to elucidate the functional signalling outcome of angiogenin focusing on its neuroprotective activities, in particular regarding ALS pathology with the aim of finding new therapies for this fatal disease.

How is pharmacy education, delivered by a new School of Pharmacy, with new undergraduate and postgraduate curricula, novel teaching and assessment strategies, reflected in student perception, performance and preparedness for professional practice, and what are the lessons for educators?

The thesis addresses three major areas; interprofessional education, competency assessment and evaluation. Quantitative methods, including the use of validated tools (Readiness for Interprofessional Learning Scale, Attitudes to Health Professionals Questionnaire, Course Experience Questionnaire), and relevant research ("MPharm: Where are we now" and the "Pharmacy Education and Accreditation Reviews" reports) were employed. Analysis of examination results and tracking of student progress was also undertaken.

An unexpected finding was that early interprofessional education was less successful than anticipated. Views on professional identity and stereotypes influenced a desire for uniprofessional education. However, online interprofessional education can facilitate learning of large groups of geographically dispersed professionals and may offer potential for postgraduate interprofessional education.

The research highlights the value of the Objective Structured Clinical Examination (OSCE) for assessing competency in pharmacy, but emphasises the importance of embedding quality assurance into the design and delivery with due regard for validity and reliability. The research indicates that it is helpful to reflect on the outcomes to determine if assessment policies are robust and credible.

Evaluation strategies are important in the development of new programmes. The introduction of the National Pharmacy Internship Programme provided a unique and previously unexplored opportunity to move beyond student perception, evaluate outcomes at entry-to-practice and benchmark the quality of education.

Some additional significant findings emerge from tracking student progress. There is no evidence that a lack of advanced second-level science education has a deleterious effect on performance. Graduate students perform better than school leavers, despite having significantly lower points on entry. their merit in considering a graudate entry programme, although it is not the currently proprosed strategy for pharmacy education in Ireland.

Pharmacy education, delivered by a new School of Pharmacy using novel and outcomes focused teaching and assessment strategies was reflected well in student perception, performance and preparedness for practice. The research is timely and important as Ireland moves towards the development of a new five-year integrated programme where the students are prepared to directly enter professional practice.

In this study it is hypothesised that the pathway expression profiles of high grade gliomas may predict response to EGFR I PDGFR pathway blockade with TKls. Specifically, it was determined if EGFR, EGFRvIII, PDGFR c-KIT and c-ABL expression together with PTEN expression and downstream activation of AKT and P70s6K predicted response to EGFR or PDGFR blockade with erlotinib, gefitinib or imatinib. The immunohistochemical (IHC) expression profiles of the EGFR I PDGFR pathway in formalin fixed paraffin embedded sections of high grade gliomas and in primary cultures derived from the same tumours were established. This facilitated the characterisation of changes introduced as a result of in vitro culturing. Next, response to treatment with TKls was determined for each treated primary culture using anti-proliferation toxicity assays. Response was correlated with the above expression profiles and with patient survival data.

Tumour expression profiles showed that no one protein of the EGFR 1 PDGFR pathway significantly influenced overall survival and that diagnostic grade remained the best prognostic indicator of outcome. The EGFR I PDGFR pathway profiles remained stable over time in patients with repeat biopsies. Comparison of pathway profiles between tumour tissue and their derived primary cultures demonstrated the introduction of significant genomic and protein differences as a result of culturing. Increased surface receptor tyrosine kinases were observed with an opposing decrease in activation of the downstream pathway proteins when the cells were grown in vitro. Despite these differences, the overall EGFR I

PDGFR pathway profile of primary cultures retained key signature characteristics reflective of the coordinated pathway disruption observed in high grade gliomas. This verified their use as a biologically representative model of high grade glioma in determining if biomarkers predicting response to tyrosine kinase inhibitors (TKls) could be identified in the EGFR I PDGFR pathway. It became apparent that although some cultured tumours responded to therapy with TKls, this response was not statistically related to the pathway proteins that were evaluated in this study.

EGFR 1 PDGFR in vitro pathway status was a poor predictor of in vitro responsiveness to molecular blockade using the TKls: erlotinib, gefitinib and imatinib. In conclusion the complexity and evasiveness of signalling pathways in high grade gliomas coupled with the possible emergence of alternate, deviant signalling pathways combine to minimise the likelihood that single agent pathway inhibitors will have a significant clinical benefit for glioma patients at this time.