Integrating receptor interactions and dynamics and Interference with endocrine disruptors in the mechanisms of action of PPAR nuclear receptors

Abstract Peroxisome Proliferator-Activated Receptors (PPARs) form a family of three nuclear receptors regulating important cellular and metabolic functions. PPARs control gene expression by directly binding to target promoters as heterodimers with the Retinoid X Receptor (RXR), and their transcriptional activity is enhanced upon activation by natural or pharmacological ligands. The binding of PPAR/RXR heterodimers on target promoters allows the anchoring of a series of coactivators and corepressors involved in promoter remodeling and the recruitment of the transcription machinery. The transcriptional output finally depends on a complex interplay between (i) the respective expression levels of PPARs, RXRs and of other nuclear receptors competing for DNA binding and RXR recruitment, (ii) the availability and the nature of PPAR and RXR ligands, (iii) the expression levels and the nature of the different coactivators and corepressors and (iv) the sequence and the epigenetic status of the promoter. Understanding how all these factors and signals integrate and fine-tune transcription remains a challenge but is necessary to understand the specificity of the physiological functions regulated by PPARs. The work presented herein focuses on the molecular mechanisms of PPAR action and aims at understanding how the interactions and mobility of the receptor modulate transcription in the physiological context of a living cell: Such observations in vivo rely on the use of engineered fluorescent protein chimeras and require the development and the application of complementary imaging techniques such as Fluorescence Recovery After Photobleaching (FRAP), Fluorescence Resonance Energy Transfer (FRET) and Fluorescence Correlation Spectroscopy (FCS). Using such techniques, PPARs are shown to reside solely in the nucleus where they are constitutively associated with RXR but transcriptional activation by ligand binding -does not promote the formation of sub-nuclear structures as observed with other nuclear receptors. In addition, the engagement of unliganded PPARs in large complexes of cofactors in living cells provides a molecular basis for their ligand-independent activity. Ligand binding reduces receptor diffusion by promoting the recruitment of coactivators which further enlarge the size of PPAR complexes to acquire full transcriptional competence. Using these molecular approaches, we deciphered the molecular mechanisms through which phthalates, a class of pollutants from the plastic industry, interfere with PPARγ signaling. Mono-ethyl-hexyl-phthalate (MEHP) binding induces the recruitment of a specific subset of cofactors and translates into the expression of a specific subset of target genes, the transcriptional output being strongly conditioned by the differentiation status of the cell. This selective PPARγ modulation induces limited adipogenic effects in cellular models while exposure to phthalates in animal models leads to protective effects on glucose tolerance and diet-induced obesity. These results demonstrate that phthalates influence lipid and carbohydrate metabolism through complex mechanisms which most likely involve PPARγ but also probably PPARα and PPARß, Altogether, the molecular and physiological demonstration of the interference of pollutants with PPAR action outlines an important role of chemical exposure in metabolic regulations. Résumé Les PPARs (Peroxisome Proliferator-Activated Receptors) forment une famille de récepteurs nucléaires qui régulent des fonctions cellulaires et métaboliques importantes. Les PPARs contrôlent l'expression des gènes en se liant directement à leurs promoteurs sous forme d'hétérodimères avec les récepteurs RXR (Retinoid X Receptor), et leur activité transcriptionnelle est stimulée par la liaison de ligands naturels ou pharmacologiques. L'association des hétérodimères PPAR/RXR avec les promoteurs des gènes cibles permet le recrutement de coactivateurs et de corépresseurs qui vont permettre le remodelage de la chromatine et le recrutement de la machinerie transcriptionnelle. Les actions transcriptionnelles du récepteur dépendent toutefois d'interactions complexes qui sont régulées par (i) le niveau d'expression des PPARs, des RXRs et d'autres récepteurs nucléaires entrant en compétition pour la liaison à l'ADN et l'association avec RXR, (ii) la disponibilité et la nature de ligands de PPAR et de RXR, (iii) les niveaux d'expression et la nature des différents coactivateurs et corépresseurs et (iv) la séquence et le marquage épigénétique des promoteurs. La compréhension des mécanismes qui permettent d'intégrer ces aspects pour assurer une régulation fine de l'activité transcriptionnelle est un défi qu'il est nécessaire de relever pour comprendre la spécificité des fonctions physiologiques régulées par les PPARs. Ce travail concerne l'étude des mécanismes d'action moléculaire des PPARs et vise à mieux comprendre comment les interactions du récepteur avec d'autres protéines ainsi que la mobilité de ce dernier régulent son activité transcriptionnelle dans le contexte physiologique des cellules vivantes. De telles observations reposent sur l'emploi de protéines fusionnées à des protéines fluorescentes ainsi que sur le développement et l'utilisation de techniques d'imagerie complémentaires telles que le FRAP (Fluorescence Recovery After Photobleaching), le FRET (Fluorescence Resonance Energy Transfer) ou la FCS (Fluorescence Corrélation Spectroscopy). En appliquant ces méthodes, nous avons pu montrer que les PPARs résident toujours dans le noyau où ils sont associés de manière constitutive à RXR, mais que l'ajout de ligand n'induit pas la formation de structures sub-nucléaires comme cela a pu être décrit pour d'autres récepteurs nucléaires. De plus, les PPARs sont engagés dans de larges complexes protéiques de cofacteurs en absence de ligand, ce qui procure une explication moléculaire à leur activité ligand-indépendante. La liaison du ligand réduit la vitesse de diffusion du récepteur en induisant le recrutement de coactivateurs qui augmente encore plus la taille des complexes afin d'acquérir un potentiel d'activation maximal. En utilisant ces approches moléculaires, nous avons pu caractériser les mécanismes permettant aux phtalates, une classe de polluants provenant de l'industrie plastique, d'interférer avec PPARγ. La liaison du mono-ethyl-hexyl-phtalate (NERF) à PPARγ induit un recrutement sélectif de cofacteurs, se traduisant par l'induction spécifique d'un sous-ensemble de gènes qui varie en fonction du niveau de différentiation cellulaire. La modulation sélective de PPARγ par le MEHP provoque une adipogenèse modérée dans des modèles cellulaires alors que l'exposition de modèles animaux aux phtalates induit des effets bénéfiques sur la tolérance au glucose et sur le développement de l'obésité. Toutefois, les phtalates ont une action complexe sur le métabolisme glucido-lipidique en faisant intervenir PPARγ mais aussi probablement PPARα et PPARß. Cette démonstration moléculaire et physiologique de l'interférence des polluants avec les récepteurs nucléaires PPAR souligne un rôle important de l'exposition à de tels composés dans les régulations métaboliques.

The Decision on Turkish EU accession in December 2002: The logic of anarchy, collective action problems and rhetorical action

Studies of the EU accession of the East and Central European Countries have stressed the importance of neo-liberal institutionalism as an explanation for Member State preferences. In this paper it is argued that Member States’ preferences over Turkish EU accession are better explained by power politics and neo-realism. It seems therefore that Turkey’s way to the EU follows another path than the East and Central Countries. Turkish accession raises the question of the EU’s role in a uni-polar world order – whether the EU should develop into an independent actor on the world stage or not. However, when it comes to the interaction among the Member States in order to decide on when to open accession negotiations with Turkey the constitutive values of the EU seriously modify the outcome that pure power politics would have let to.

The New EU’s External Action: A significant step forward towards the EU’s single voice?ction problems and rhetorical action

The aim of this paper is to analyse the main agreements on the EU’s External Action agreed within the European Convention and the IGC taking into account why, how and who reached the consensus on them. In other words, this paper will explore the principles followed in order to improve the instruments of the EU’s External Action such as authority, coherence, visibility, efficiency and credibility.

Tri-sector partnerships in social entrepreneurship: discourse and practice of the actors from the circles of action and reflection

This article discusses the construction of tri-sector partnerships in three projects conducted in Brazil in different fields of intervention of public policy (access to water, basic education and performance of boards of rights of children and adolescents). Collaborative articulations involving the players from three sectors (the State, civil society and the market) are practices that are little studied in the Brazilian and even in the international context, as tri-sector partnerships are rare, despite the proliferation of lines of discourse in support of alliances between governments and civil society or between companies and NGOs in the management of public policy. As a research strategy, this study resorted to cooperative inquiry, a method that involves breaking down the boundaries between the subjects and the objects of the analysis. Besides working toward a better understanding of the challenges of building tri-sector partnerships in the Brazilian context, the article also tries to show the relevance to public policy studies of investigative methods based on the subjects studied, as a means of developing an understanding of the practices, lines of discourse and dilemmas linked to social action in social programs.

La Gestió policial als esdeveniments públics en espais urbans a l'Àfrica: situació actual i perspectives = Policing public events in urban spaces in Africa : an analysis of the current situation and a perspective on follow-up action

L’informe que es presenta en aquest llibre és el resultat d’un nou acord de col·laboració entre el Programa de les Nacions Unides per als Assentaments Humans (ONU-Habitat) i l’Institut de Seguretat Pública de Catalunya, impulsat amb l’objectiu de millorar la seguretat en esdeveniments públics en els espais urbans a l’Àfrica. La fase pilot es va dur a terme el 2010, durant els dos seminaris de formació realitzats a Mollet del Vallès (Barcelona) com a part de la Plataforma Policia per al Desenvolupament Urbà (PPUD). En aquest informe es descriuen els orígens i l’estat de la iniciativa i resumeix els resultats. També s’inclouen algunes recomanacions per a millorar la seguretat d’esdeveniments públics. Font d'informació: http://www.onuhabitat.org.

Use of drugs with more than a twenty-four-hour duration of action

AIM: To assess compliance with a drug regimen of two doses a day compared with one a day. PATIENTS AND METHODS: A prospective crossover study was set up in a general practice environment to compare compliance on a drug regimen of once a day versus twice a day. Data were collected by electronic monitoring in 113 patients with hypertension or angina pectoris. All patients were prescribed slow-release nifedipine twice a day during the first month and then crossed to a single daily dose of amlodipine for another month. RESULTS: Compliance, defined as the proportion of days on which the correct dose was taken, improved in 30% of patients (95% confidence interval 19-41%; P < 0.001) when the patients were switched from twice a day to once a day, but at the same time there was a 15% increase (95% confidence interval 5-25%; P < 0.02) in the number of patients with one or more no-dose days. Approximately 8% of patients displayed low compliance, irrespective of the dose regimen. Actual dose intervals were used to estimate the extent and timing of periods with unsatisfactory drug activity for various hypothetical drug durations of action. CONCLUSIONS: The apparent advantage of a single daily dose in terms of compliance appears to be clinically meaningful only when the duration of activity extends beyond the dose interval in all patients.

Training effects with the observer-rated cognitive errors and the coping action patterns scales

The use of observer-rated scales requires that raters be trained until they have become reliable in using the scales. However, few studies properly report how training in using a given rating scale is conducted or indeed how it should be conducted. This study examined progress in interrater reliability over 6 months of training with two observer-rated scales, the Cognitive Errors Rating Scale and the Coping Action Patterns Rating Scale. The evolution of the intraclass correlation coefficients was modeled using hierarchical linear modeling. Results showed an overall training effect as well as effects of the basic training phase and of the rater calibration phase, the latter being smaller than the former. The results are discussed in terms of implications for rater training in psychotherapy research.

Iowa Department of Human Right - Community Action, Agency Performance Plan

Annual Report, Agency Performance Plan

Iowa Civil Rights Commission, Agency Performance Plan and Action Plan

Annual Report, Agency Performance plan

Lottery Action, July 8-21, 2002, Vol. 9 no. 13

Iowa Lottery Authority Retailer Information Newsletter