Insulin-like growth factor receptor activity in cancer biology & therapy responses

The Insulin-like Growth Factor 1 Receptor (IGF-1R) has an essential function in normal cell growth and in cancer progression. However, anti-IGF-1R therapies have mostly been withdrawn from clinical trials owing to a lack of efficacy and predictive biomarkers. IGF-1R activity and signalling in cancer cells is regulated by its C-terminal tail, and in particular, by a motif that encompasses tyrosines 1250 and 1251 flanked by serines 1248 and 1252 (1248- SFYYS-1252). Mutation of Y1250/1251 greatly reduces IGF-1-promoted cell migration, interaction with the scaffolding protein RACK1 in the context Integrin signalling, and IGF- 1R kinase activity. Here we investigated the phosphorylation of the SFYYS motif and characterise the conditions under which this motif may be phosphorylated under. As phosphorylated residues, the SFYYS motif may also serve to recruit interacting proteins to the IGF-1R. To this end we identified a novel IGF-1R interacting partner which requires phosphorylated residues in the SFYYS motif to interact with the IGF-1R. This interaction was found to be IGF-1-dependent, and required the scaffold protein RACK1. The interaction of this binding protein with the IGF-1R likely functions to promote maximal phosphorylation of Shc and ERK in IGF-1-stimulated cell migration, and may be important for IGF-1 signalling in cancer cells. Lastly, we have investigated possible kinases that may confer resistance or sensitivity to the IGF-1R kinase inhibitor BMS-754807. In this screen we identified ATR as a mediator of resistance and showed that suppression or chemical inhibition of ATR synergised with BMS-754807 to reduce colony formation. This work has contributes to our understanding of IGF-1R kinase regulation and signalling and suggests that administration of anti-IGF-1R drugs with ATR inhibitors may have therapeutic benefit.

A literary occupation: responses of German writers in service in occupied Europe

This thesis examines the literary output of German servicemen writers writing from the occupied territories of Europe in the period 1940-1944. Whereas literary-biographical studies and appraisals of the more significant individual writers have been written, and also a collective assessment of the Eastern front writers, this thesis addresses in addition the German literary responses in France and Greece, as being then theatres of particular cultural/ideological attention. Original papers of the writer Felix Hartlaub were consulted by the author at the Deutsches Literatur Archiv (DLA) at Marbach. Original imprints of the wartime works of the subject writers are referred to throughout, and citations are from these. As all the published works were written under conditions of wartime censorship and, even where unpublished, for fear of discovery written in oblique terms, the texts were here examined for subliminal authorial intention. The critical focus of the thesis is on literary quality: on aesthetic niveau, on applied literary form, and on integrity of authorial intention. The thesis sought to discover: (1) the extent of the literary output in book-length forms. (2) the auspices and conditions under which this literary output was produced. (3) the publication history and critical reception of the output. The thesis took into account, inter alia: (1) occupation policy as it pertained locally to the writers’ remit; (2) the ethical implications of this for the writers; (3) the writers’ literary stratagems for negotiating the constraints of censorship.

Pregnancy-specific glycoprotein function, conservation and receptor investigation

Pregnancy-specific glycoproteins (PSGs) are highly glycosylated secreted proteins encoded by multi-gene families in some placental mammals. They are carcinoembryonic antigen (CEA) family and immunoglobulin (Ig) superfamily members. PSGs are immunomodulatory, and have been demonstrated to possess antiplatelet and pro-angiogenic properties. Low serum levels of these proteins have been correlated with adverse pregnancy outcomes. Objectives: Main research goals of this thesis were: 1). To attempt to replicate previously reported cytokine responses to PSG-treatment of immune cells and subsequently to investigate functionally important amino acids within PSG1. 2). To determine whether candidate receptor, integrin αvβ3, was a binding partner for PSG1 and to investigate whether PSG1 possessed functionality in a leukocyte-endothelial interaction assay. 3). To determine whether proteins generated from recently identified putative PSG genes in the horse shared functional properties with PSGs from other species. Outcomes: 1). Sequential domain deletion of PSG1 as well as mutation of conserved residues within the PSG1 Ndomain did not affect PSG1-induced TGF-β1. The investigated response was subsequently found to be the result of latent TGF-β1 contaminating the recombinant protein. Protein further purified by SEC to remove this showed no induction of TGF-β1. The most N-terminal glycosylation site was demonstrated to have an important role in PSG N domain secretion. PSG1 attenuated LPS-induced IL-6 and TNF-α. Investigations into signalling underpinning this proved inconclusive. 2). Integrin αvβ3 was identified as a novel PSG1 receptor mediating an as yet unknown function. Preliminary investigations into a role for PSGs as inhibitors of leukocyte endothelial interactions showed no effect by PSG1. 3). Horse PSG protein, CEACAM49, was shown to be similarly contaminated by latent TGF-β1 particle and once removed did not demonstrate TGF-β1 release. Interestingly horse PSG did show anti-platelet properties through inhibition of the plateletfibrinogen interaction as previously published for mouse and human PSGs.

The student bullying of teachers in Irish second level schools: exploring the influence of historical low state intervention in education on the development of contemporary policy responses in Ireland

The student bullying of teachers (SBT) is a distinct and complex form of bullying with a multiplicity of diverse, changeable and intersecting causes which is experienced by and affects teachers in a variety of ways. SBT is both a national and an international phenomenon which is under-recognised in academic, societal and political spheres, resulting in limited conceptual understanding and awareness of the issue. This study explores teachers’ experiences of SBT behaviours in Irish second level schools as well as teachers’ perceptions regarding training, policies and supports in Ireland to address the issue. Specifically, the study seeks to explore the influence of historical low State intervention in education on contemporary policies and supports to deal with SBT in Ireland. A mixed methods approach involving a survey of 531 second level school teachers and 17 semi-structured interviews with teachers, Year Heads and representatives from teacher trade unions and school management bodies was employed to collect and analyse data. Findings indicate that SBT behaviours are prevalent in many forms in Irish second level schools. The hidden nature of the phenomenon has simultaneously contributed to and is reinforced by limited understanding of the issue as well as teachers’ reluctance to disclose their experiences. Findings reveal that teachers perceive the contemporary policies, training and support structures in Ireland to be inadequate in equipping them to effectively deal with SBT. State intervention in addressing SBT behaviours to date, has been limited, therefore many teachers are forced to respond to the issue based on their own initiatives and assumptions rather than from an informed critically reflective approach, supported by national guidelines and sufficient State investment. This has resulted in a piecemeal, un-coordinated and ad-hoc approach to SBT in Irish schools both in terms of teachers’ management of SBT behaviours and with respect to the supports extended to staff. The potential negative consequences of SBT behaviours on teachers’ wellbeing and professional performance and thus, on the education system itself, underlines the need for a strategic, evidence-based, resourced and integrated approach which includes, as a pivotal component, consultation with teachers, whose contribution to the process is crucial.