Insulin signalling is an essential process in humans by which the level of plasma glucose is maintained within the physiologically healthy range. Insulin activates the phosphoinositide 3 kinase (PI3K) signalling pathway that generates the phospholipid messenger PtdIns(3,4,5)P3, which in turn enhances the activity of two important proteins, AKT and Rac1. This then leads to increase the presence of the glucose transporter 4 (GLUT4) at the plasma membrane that enhances the intake of glucose, particularly in skeletal muscle cells and adipocytes. Insulin signalling also triggers interconversion of several other phosphoinositides (PIs) which play pivotal roles in different steps of glucose regulation. PtdIns5P is an important PI that is robustly increased after insulin treatment in the skeletal muscle cell line, L6 myotubes. Many of PtdIns5P`s functions are not fully understood. To gain more knowledge of the role of PtdIns5P in insulin signalling in muscle cells, the PtdIns5P kinase phosphatidylinositol-5-phosphate 4-kinase a (PIP4K2a) was over-expressed in L6 myotubes as a way of removing PtdIns5P, and the consequences in insulin signalling were studied. Although PtdIns5P is converted by PIP4K2a to PtdIns(4,5)P2 which is a precursor of the potent PI PtdIns(3,4,5)P3, previous studies revealed that the increase in PtdIns(3,4,5)P3 induced by insulin in control cells is diminished in cells overexpressing PIP4K2a, for unknown reasons. Additionally, although the phosphorylation of the serine/threonine protein kinase AKT was not affected in these L6 cells, glucose uptake was attenuated. The current study investigates the possible causes of attenuating glucose uptake in PIP4K overexpressing myotubes by examining the small GTPase Rac1 which plays an important role in the cytoskeleton re-arrangement that is necessary for GLUT4 translocation. Furthermore, the possible roles of PI phosphatases that may cause the disturbance on the levels of PIs in response to insulin were evaluated. Additionally, the potential role of PtdIns5P in Rac1 activation in L6 myotubes was further investigated by delivering synthetic PtdIns5P using a carrier-based delivery approach. The results showed that the attenuation of glucose uptake documented in previous studies occurred as a result of a defect in the process of translocating GLUT4 from intracellular storage to the plasma membrane. Rac1 activity was significantly reduced in cells expressing PIP4K2a. Quantifying the level of PIs suggested that PIP4K2a expression increases the removal of PtdIns(3,4,5)P3 by the PI 5-phosphatase, SKIP. Silencing the expression of SKIP by siRNA restored the level of PtdIns(3,4,5)P3 but Rac1 activity and the attenuation GLUT4 translocation were not rescued possibly as a result of removing PtdIns5P by PIP4K2a. On the other hand, exogenous delivery of PtdIns5P in L6 myotubes activates both Rac1 and GLUT4 translocation in the absence of insulin. However, activating GLUT4 translocation by the exogenous PtdIns5P requires PI3K activity since redistribution of GLUT4 to the plasma membrane is inhibited by the PI3K inhibitor, wortmannin. Removing PtdIns5P reduces Rac1 activity and stimulates SKIP that inhibits PtdIns(3,4,5)P3 increase which attenuates GLUT4 translocation and hence glucose uptake. These results emphasise the critical role played by PtdIns5P which seems to serve as a regulator of insulin signalling, both directly and/or by regulating other enzymes involved in the metabolism of PIs.

The control of phosphatidylinositol 3, 4, 5-trisphosphate [PtdIns(3,4,5)P3] level depends on the activities of both PI kinase and PtdIns(3,4,5)P3 phosphatases: 5-phosphatase like SHIP1 and SHIP2, and 3-phosphatase like PTEN. The ubiquitous SH2 domain containing inositol 5-phosphatase SHIP2 contains both a series of protein interacting domains and the ability to dephosphorylate PtdIns(3,4,5)P3. Previous reports obtained in SHIP2 deficient mice have shown that SHIP2 is involved in the control of insulin sensitivity and reducing weight gain on fatty diet. Since SHIP2 is a lipid phosphatase as well as a docking protein, the initial aim that emerged in the lab was to measure the inositol lipid levels in SHIP2 +/+ and deficient cells and compare the levels of 3-phosphoinositides PtdIns(3,4,5)P3 and PtdIns(3,4)P2. At first, we developed mouse embryonic fibroblasts (MEF) as a cellular model. Amongst various stimuli tested, surprisingly, only serum showed an obvious difference in terms of PtdIns(3,4,5)P3 level. This lipid was significantly up regulated in SHIP2 -/- cells but only after short-term (i.e. 5-10 min) incubation with serum. The difference in PtdIns(3,4,5)P3 levels in heterozygous fibroblast cells was intermediate between the +/+ and -/- cells. Serum stimulated PI3K activity appeared to be comparable between +/+ and -/- cells. Moreover, PKB, but not MAP kinase activity, was also potentiated in SHIP2 deficient cells stimulated by serum. The up regulation of PKB activity in serum stimulated cells was totally reversed in the presence of the PI3K inhibitor LY-294002, in both +/+ and -/- cells. Reactive oxygen species (ROS) have emerged as physiological mediators of many cellular responses. H2O2 mimics some effects of insulin in a number of cell culture systems. It also inactivates tyrosine phosphatase activities including PTEN. In addition, in Swiss 3T3 fibroblasts, Gray et al reported that exposure of the cells to H2O2 resulted a huge increase in PtdIns(3,4)P2 level. The authors suspected that the effect was attributed to a inositol 5-phosphatase activity. We thus exposed our cells to H2O2 in order to address the question of the role of SHIP2 in response to oxidative stress. We worked on the same SHIP2 MEF model, stimulated by H2O2: at 15 min, PtdIns(3,4,5)P3 was markedly increased in SHIP2 -/- cells as compared to +/+ cells. In contrast, no significant increase in PtdIns(3,4)P2 could be detected at 15 or 120 min incubation of the cells with H2O2 (0.6 mM). PKB activity was upregulated in SHIP2 -/- cells in response to H2O2 and therefore follows the regulation of PtdIns(3,4,5)P3. As for serum, the PI3K activity appeared to be comparable between +/+ and -/- cells. The levels of PTEN and type I 4-phosphatase [an enzyme that acts on PtdIns(3,4)P2] remained unchanged between the two types of cells. SHIP2 add back experiments in SHIP2 -/- cells confirm its critical role in the control of PtdIns(3,4,5)P3 level in response to H2O2: the decrease in PtdIns(3,4,5)P3, observed in SHIP2 expressing cells, was no longer seen in cells infected with a catalytic mutant of this enzyme.

This essay aims to analyse the form of two radio documentary shows and compare them to, what I mean are a traditional, Swedish radio documentary form. The radio shows are Radiolab distributedby National Public Radio and WNYC, Övernaturligt (supernatural) produced by TredjeStatsmakten Media AB and P3 Dokumentär (P3 Documentary) distributed by Sveriges Radio. Alarge part of the essay is dedicated to develop a method to analyse form in specific radio shows. Mymethod consists of two parts:1. A defining of sounds in a spreadsheet that map out the different sound elements over time.2. An analysis of the radio documentaries using the spreadsheet, text-analytic tools, rhetoric toolsand an holistic view of how different stories connect.My essay shows how these two radio documentary shows use so called layers and the changing ofroles between the presenter and the reporter to replicate a live show in closeness to the audience andin turn, how this may affect the presentation of their journalistic content. I also show how theseradio shows use illustrative and side stories to build up the main story of the show.

Events that violate expectations are biologically significant and accordingly elicit various physiological responses. We investigated the functional relationship between three of these responses: the P300, the Novelty P3 and the pupil dilation response (PDR), with a particular focus on their co-variance with reaction time and measures of subsequent memory. In a modified Novelty P3 oddball paradigm, participants semantically categorized a sequence of stimuli including (1) words of a frequent category, (2) words of an infrequent category (14% of the trials) and (3) pictures of the frequent category (14% of the trials). The Novelty P3 oddball task was followed by a recall- and a recognition test. Larger amplitudes of the P300, identified by a spatial principal component analysis (PCA), were associated with enhanced subsequent recall as well as faster reaction times during the recognition test, suggesting a close relationship between the cognitive process indexed by the P300 and memory encoding. The PDR was larger for infrequents (which required a response switch) than both frequents and pictures (which did not require a switch). Furthermore, its latency was correlated with reaction time on the same trial and with reaction time on the immediately following trial. There was only weak evidence for a correlation with subsequent memory, suggesting that the cognitive process associated with the PDR might be a direct link in the stimulus-response stream. Larger Novelty P3 amplitudes were associated with both faster reaction times on the same trial and stronger memory traces, suggesting that its amplitude might index resource allocation. These findings suggest that each of the physiological responses carries a distinct functional significance in detecting, processing, or responding to novel events, and we discuss the findings in the light of the prevalent theories of the functional significance of each response.

While the outright privatization of water services has declined globally, it has been replaced with public-private partnerships (P3s) in the government procurement and delivery of water services, and increasingly at the local level. This research finds that such initiatives are on the rise in Canada, and considering the overall record of failure that has amounted from varied types of water privatization thus far, it seeks to analyze this expanding waterscape from a critical perspective. More specifically, by historically situating the privatization of Canadian municipal water in a political-economic context that identifies its relation to contemporary (neoliberal) capitalism, this research examines how the focused state commitment to water P3s is indicative of the processes of neoliberalization. I argue that regulatory and budgetary changes since the economic crisis of 2008 have formed an institutionalized policy apparatus that essentially forces needy municipalities into long-term contracts with private firms, therefore establishing sustained sites for capital accumulation. This thesis concludes with a discussion concerning the implications that such a “partnership” will have for municipal autonomy, organized labour, and the environment, particularly in light of the intensifying state focus on international free trade. / Graduate / 0626 / 0629

abstract: Public Private Partnerships (PPP) have been in use for years in the United Kingdom, Europe, Australia and for a shorter time here in the United States. Typical PPP infrastructure projects include a multi-year term of operation in addition to constructing the structural features to be used. Early studies are proving PPP delivery methods to be effective at construction cost containment. An examination of the key elements that constitute the early stage negotiation reveal that there is room for negotiation created by the governing documentation while maintaining a competitive environment that brings the best value available to the Public entity. This paper will examine why PPP's are effective during this critical construction period of the facilities life cycle. It is the intent of this study to examine why the features and outcomes of more or less negotiation and the degree of rigor associated with it. / Dissertation/Thesis / M.S. Construction 2012

La jouissance est un des champs dont la psychanalyse fait usage pour rendre compte de ce qui pourrait expliquer le boitement structurel de la condition humaine, son paradoxe fondamental. Partant d'une épistémologie transcendantale, nous envisageons que, bien que le corps et la psyché ne revêtent pas les mêmes réalités, ils ont certains points d'accroche dont fait partie la jouissance et, bien que le corps ne détermine pas le psychisme, il le contraint. Il est alors proposé ici une hypothèse neuropsychanalytique de la jouissance, l'articulant autour du corps et de l'histoire. La jouissance serait la marque de ce qui a fait événement pour le sujet. Cette description permet de relier ce concept à la neurophysiologie des circuits dopaminergiques qui marquent physiologiquement tant la dimension pulsionnelle, que la dimension événementielle et donc historique.À partir de cette conceptualisation, un paradigme expérimental a été constitué afin d'investiguer cette possibilité d'une accroche entre le psychique et le biologique chez des sujets dépendants, lorsqu'ils sont confrontés à leur objet d'addiction. Concrètement, nous 'osons' l'hypothèse que plus un sujet manifeste d'indices repérables d'une mobilisation pulsionnelle dans son discours libre amorcé par l’objet addictif (dans ce cas-ci, la cigarette pour des fumeurs), plus il aura le geste rapide et sûr pour l’attraper ou des difficultés à inhiber son appétence vers cet objet. Ceci est investigué à travers un outil constitué à partir de la théorie psychanalytique d’une part, et à travers deux tâches de type Go-NoGo (évaluées par des mesures comportementales –temps de réaction et erreurs- et électrophysiologiques -la N2 et la P3, deux marqueurs considérés comme reflétant la signature des processus attentionnels et des biais d’inhibition-, d’autre part. La particularité du paradigme, outre cet usage d’outils issus de champs distincts, réside dans le fait que, à aucun moment, ce n’est le sens du discours des sujets, qui est investigué. L’outil psychanalytique évalue effectivement la façon dont les sujets s’expriment, et ce sont des juges naïfs (à l’égard de l’étude et de la psychanalyse) qui évaluent leur discours. Nos résultats mettent en évidence que les personnes dépendantes manifestent une tendance à l’impulsivité de façon générale. Mais, lorsque leur objet d’addiction est en place de support attentionnel, les sujets démontrent une augmentation significative de leurs performances et de l’amplitude de la P3, signe d’un investissement attentionnel. Les résultats des associations libres révèlent que la forme du discours des fumeurs est évaluée comme plus transgressive (comparativement aux non-fumeurs). Enfin, on retrouve une corrélation entre les indices psychiques (la transgression) et neurophysiologiques (la latence de la NoGo-N2). En conclusion, il semble que l’addiction dépasse la simple réalité pharmacologique de l’effet du toxique puisqu’il est repéré ici que l’effet de stimulant cognitif (manifeste par l’amélioration des performances) est induit par la simple représentation de la cigarette et non par la consommation de celle-ci. Ensuite, au regard de notre proposition épistémologique, nous semblons avoir réussi, sur base d’une lecture rapprochée des séminaires de Lacan, à construire un outil qui distingue de façon cohérente, consistante et valide une dimension pulsionnelle d’une dimension affective (valencée) dans ce qui peut s’entendre. Grâce à cet outil, nous avons trouvé des corrélations entre certains de ces marqueurs psychiques et certains paramètres physiologiques - les paramètres corrélant étant précisément des plus probants et la corrélation s’avérant être dans le sens prédit par notre construction hypothétique. Sur ce 'succès’, nous osons une conclusion épistémologique plus large qui propose que si l’univers du représentationnel, c'est-à-dire, du mental, est bien accroché au niveau de repères physiologiques, ceci n’est démontrable sur un plan mental que si l’on se borne à la forme des contenus mentaux – et, qui plus est, et de façon spéculative, en évitant toute hérméneutique au niveau des contenus mentaux. / Doctorat en Sciences psychologiques et de l'éducation / info:eu-repo/semantics/nonPublished

This dissertation focuses on the findings of a multi-case study centered around two public-private partnership (P3s) projects in Ontario: the Royal Ottawa Hospital and the Brampton Civic Hospital. Partnerships have become particularly important as a part of infrastructure procurement for all levels of government in Canada. While infrastructure public-private partnerships have grown in popularity, they remain a controversial means of procuring public assets. Considerable questions remain as to whether the mechanisms related to partnerships with the private sector represent a sufficient response to the challenges facing health care systems. As such, major avenues exist for contributions in the form of evidence-based examinations to the field of knowledge pertaining to hospital procurement. A body of research literature and review of public-private partnerships, including those with a particular focus on health-sector projects in Canada, has grown over the past two decades. This study contributes an analysis of the relationship between public and private partners to that literature, utilizing a conceptual lens developed out of complex evolutionary systems theory. The study is intended to examine the capacity of public managers entering into a public-private partnership arrangement to successfully activate agents and utilize the skill and knowledge of these agents, conduct joint fact finding and consensus building, and better understand how they have arranged and organized joint interactions between the public and private sector. The two case studies demonstrate the key inputs into the decision-making processes for what were formative health infrastructure partnership projects in Ontario, providing an assessment of the degree to which the government of Ontario was successful in managing a cooperative decision-making process that stressed inclusion and horizontal steering. Results find that public managers struggled in some ways to leverage an effective horizontal management style and engage in smart interventions to utilize expert knowledge to address knowledge gaps, contributing to stagnating negotiations and driving up transaction costs for the projects. Some noteworthy successes were experienced in the case of the Royal Ottawa Hospital in leveraging private-sector knowledge to develop performance metrics, and the approach to stakeholder engagement in this case presents positive lessons-learned for future P3 infrastructure projects.

Hur ser svenska nyhetstexters rapportering om fallet Yasin och P3-Guld ut? Studien undersöker vilka diskurser som materialiseras i Aftonbladet, Expressen och Dagens Nyhetersrapportering från händelsen via Norman Faircloughs kritiska diskursanalys. Vidare undersöks och analyseras fallet Yasin och P3-Guld som en del av cancel-kultur med utgångspunkt i Michel Foucaults teori om brott och straff. Studien kom fram till att nyhetsrapporteringen innehöll fyra centrala teman: kriminalitet, bojkott, gangsterapp och konstnärlig frihet. Därefter analyserades materialet interdiskursivt och intertextuellt där resultatet visar på att nyhetstexterna och dess diskurser bygger på: den juridiska diskursen, tysta/lyssna diskursenoch en politisk diskurs. Nyhetstexterna kunde även analyseras via Michel Foucaults teori om övervakning och straff och studien fann tendenser i hur rapporteringen kan kopplas till cancel-kultur

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