Génomique fonctionnelle des cellules corticotropes hypophysaires : contrôle génétique de la gestion systémique des stress

L'axe hypothalamo-hypophyso-surrénalien (HPA) permet de maintenir l'homéostasie de l'organisme face à divers stress. Qu'ils soient de nature psychologique, physique ou inflammatoire/infectieux, les stress provoquent la synthèse et la libération de CRH par l'hypothalamus. Les cellules corticotropes hypophysaires perçoivent ce signal et en réaction, produisent et sécrètent l'ACTH. Ceci induit la synthèse des glucocorticoïdes (Gc) par le cortex surrénalien; ces stéroïdes mettent le système métabolique en état d’alerte pour la réponse au stress et à l’agression. Les Gc ont le rôle essentiel de contrôler les défenses de l'organisme, en plus d'exercer une rétro-inhibition sur l'axe HPA. L'ACTH est une petite hormone peptidique produite par le clivage d'un précurseur: la pro-opiomélanocortine (POMC). À cause de sa position critique dans la normalisation de l'homéostasie, le contrôle transcriptionnel du gène Pomc a fait l'objet d'études approfondies au cours des dernières décennies. Nous savons maintenant que la région promotrice du gène Pomc permet une expression ciblée dans les cellules POMC hypophysaires. L'étude du locus Pomc par des technologies génomiques m'a permis de découvrir un nouvel élément de régulation qui est conservé à travers l'évolution des mammifères. La caractérisation de cet enhancer a démontré qu'il dirige une expression restreinte à l'hypophyse, et plus particulièrement dans les cellules corticotropes. De façon intéressante, l'activité de cet élément dépend d'un nouveau site de liaison recrutant un homodimère du facteur de transcription Tpit, dont l'expression est également limitée aux cellules POMC de l'hypophyse. La découverte de cet enhancer ajoute une toute nouvelle dimension à la régulation de l'expression de POMC. Les cytokines pro-inflammatoires IL6/LIF et les Gc sont connus pour leur antagonisme sur la réaction inflammatoire et sur le promoteur Pomc via l'action des facteurs de transcription Stat3 et GR respectivement. L'analyse génomique des sites liés ii par ces deux facteurs nous a révélé une interrelation complexe et a permis de définir un code transcriptionnel entre ces voies de signalisation. En plus de leur action par interaction directe avec l’ADN au niveau des séquences régulatrices, ces facteurs interagissent directement entre eux avec des résultats transcriptionnels différents. Ainsi, le recrutement de GR par contact protéine:protéine (tethering) sur Stat3 étant lié à l'ADN provoque un antagonisme transcriptionnel. Inversement, le tethering de Stat3 sur GR supporte une action synergique, tout comme leur co-recrutement à l'ADN sur des sites contigus ou composites. Lors d'une activation soutenue, ce synergisme entre les voies IL6/LIF et Gc induit une réponse innée de défense cellulaire. Ainsi lors d'un stress majeur, ce mécanisme de défense est mis en branle dans toutes les cellules et tissus. En somme, les travaux présentés dans cette thèse définissent les mécanismes transcriptionnels engagés dans le combat de l'organisme contre les stress. Plus particulièrement, ces mécanismes ont été décrits au niveau de la réponse globale des corticotropes et du gène Pomc. Il est essentiel pour l'organisme d'induire adéquatement ces mécanismes afin de faire face aux stress et d'éviter des dérèglements comme les maladies inflammatoires et métaboliques.

Cortisol salivaire chez des mères adolescentes et leur nourrisson : exploration de quelques corrélats psychopathologiques et comportementaux

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Cortisol salivaire chez des mères adolescentes et leur nourrisson : exploration de quelques corrélats psychopathologiques et comportementaux

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Mice behaviorally selected for high Pavlovian fear memory exhibit fear generalization, HPA-axis alterations and increased basal activity in cortico-limbic circuits

Biological factors underlying individual variability in fearfulness and anxiety have important implications for stress-related psychiatric illness including PTSD and major depression. Using an advanced intercross line (AIL) derived from C57BL/6 and DBA/2J mouse strains and behavioral selection over 3 generations, we established two lines exhibiting High or Low fear behavior after fear conditioning. Across the selection generations, the two lines showed clear differences in training and tests for contextual and conditioned fear. Before fear conditioning training, there were no differences between lines in baseline freezing to a novel context. However, after fear conditioning High line mice demonstrated pronounced freezing in a new context suggestive of poor context discrimination. Fear generalization was not restricted to contextual fear. High fear mice froze to a novel acoustic stimulus while freezing in the Low line did not increase over baseline. Enhanced fear learning and generalization are consistent with transgenic and pharmacological disruption of the hypothalamic-pituitary-adrenal axis (HPA-axis) (Brinks, 2009, Thompson, 2004, Kaouane, 2012). To determine whether there were differences in HPA-axis regulation between the lines, morning urine samples were collected to measure basal corticosterone. Levels of secreted corticosterone in the circadian trough were analyzed by corticosterone ELISA. High fear mice were found to have higher basal corticosterone levels than low line animals. Examination of hormonal stress response components by qPCR revealed increased expression of CRH mRNA and decreased mRNA for MR and CRHR1 in hypothalamus of high fear mice. These alterations may contribute to both the behavioral phenotype and higher basal corticosterone in High fear mice. To determine basal brain activity in vivo in High and Low fear mice we used manganese-enhanced magnetic resonance imaging (MEMRI). Analysis revealed a pattern of basal brain activity made up of amygdala, cortical and hippocampal circuits that was elevated in the High line. Ongoing studies also seek to determine the relative balance of excitatory and inhibitory tone in the amygdala and hippocampus and the neuronal structure of its neurons. While these heterogeneous lines are selected on fear memory expression, HPA-axis alterations and differences in hippocampal activity segregate with the behavioral phenotypes. These differences are detectable in a basal state strongly suggesting these are biological traits underlying the behavioral phenotype (Johnson et al, 2011).

The relationship between stress, HPA axis functioning and brain structure in first episode psychosis over the first 12 weeks of treatment

Stress and abnormal hypothalamic-pituitary-adrenal axis functioning have been implicated in the early phase of psychosis and may partly explain reported changes in brain structure. This study used magnetic resonance imaging to investigate whether biological measures of stress were related to brain structure at baseline and to structural changes over the first 12 weeks of treatment in first episode patients (n=22) compared with matched healthy controls (n=22). At baseline, no significant group differences in biological measures of stress, cortical thickness or hippocampal volume were observed, but a significantly stronger relationship between baseline levels of cortisol and smaller white matter volumes of the cuneus and anterior cingulate was found in patients compared with controls. Over the first 12 weeks of treatment, patients showed a significant reduction in thickness of the posterior cingulate compared with controls. Patients also showed a significant positive relationship between baseline cortisol and increases in hippocampal volume over time, suggestive of brain swelling in association with psychotic exacerbation, while no such relationship was observed in controls. The current findings provide some support for the involvement of stress mechanisms in the pathophysiology of early psychosis, but the changes are subtle and warrant further investigation.

Stress, the hippocampus and the HPA axis: Implications for the development of psychotic disorders

The experience of stress is commonly implicated in models of the onset of psychotic disorders. However, prospective studies investigating associations between biological markers of stress and the emergence of psychotic disorders are limited and inconclusive. One biological system proposed as the link between the psychological experience of stress and the development of psychosis is the Hypothalamic-Pituitary-Adrenal (HPA) axis. This paper summarizes and discusses evidence supporting a role for HPA-axis dysfunction in the early phase of schizophrenia and related disorders. METHOD A selective review of psychiatric and psychological research on stress, coping, HPA-axis, the hippocampus and psychotic disorders was performed, with a particular focus on the relationship between HPA-axis dysfunction and the onset of psychotic disorders. RESULTS Individual strands of past research have suggested that the HPA-axis is dysfunctional in at least some individuals with established psychotic disorders; that the hippocampus is an area of the brain that appears to be implicated in the onset and maintenance of psychotic disorders; and that an increase in the experience of stress precedes the onset of a psychotic episode in some individuals. Models of the onset and maintenance of psychotic disorders that link these individual strands of research and strategies for examining these models are proposed in this paper. CONCLUSIONS The current literature provides some evidence that the onset of psychotic disorders may be associated with a higher rate of stress and changes to the hippocampus. It is suggested that future research should investigate whether a relationship exists between psychological stress, HPA-axis functioning and the hippocampus in the onset of these disorders. Longitudinal assessment of these factors in young people at 'ultra' high risk of psychosis and first-episode psychosis cohorts may enhance understanding of the possible interaction between them in the early phases of illness.

PVA-SSA-HPA Mixed-Matrix-Membrane Electrolytes for DMFCs

Stabilized forms of heteropolyacids (HPAs), namely phosphomolybdic acid (PMA), phosphotungstic acid (PTA), and silicotungstic acid (STA), are incorporated into poly (vinyl alcohol) (PVA) cross-linked with sulfosuccinic acid (SSA) to form mixed-matrix membranes for application in direct methanol fuel cells (DMFCs). Bridging SSA between PVA molecules not only strengthens the network but also facilitates proton conduction in HPAs. The mixed-matrix membranes are characterized for their mechanical stability, sorption capability, ion-exchange capacity, and wetting in conjunction with their proton conductivity, methanol permeability, and DMFC performance. Methanol-release kinetics is studied ex situ by volume-localized NMR spectroscopy (employing point-resolved spectroscopy'') with the results clearly demonstrating that the incorporation of certain inorganic fillers in PVA-SSA viz., STA and PTA, retards the methanol-release kinetics under osmotic drag compared to Nafion, although PVA-SSA itself exhibits a still lower methanol permeability. The methanol crossover rate for PVA-SSA-HPA-bridged-mixed-matrix membranes decreases dramatically with increasing current density rendering higher DMFC performance in relation to a DMFC using a pristine PVA-SSA membrane. A peak power density of 150 mW/cm(2) at a load current density of 500 mA/cm(2) is achieved for the DMFC using a PVA-SSA-STA-bridged-mixed-matrix-membrane electrolyte. (C) 2010 The Electrochemical Society. [DOI: 10.1149/1.3465653] All rights reserved.

The River Axe SAP Consultation Document

This is the River Axe Salmon Action Plan Consultation document produced by the Environment Agency in 2003. The report pays attention on the external consultation of the River Axe Salmon Action Plan (SAP). This strategy represents an entirely new approach to salmon management within the UK and introduces the concept of river-specific salmon spawning targets as a salmon management tool. This document is part of a national initiative to produce action plans for the management of all the main salmon rivers of England and Wales by 2003. The aim of this plan is (i) to assess the status of the salmon stocks and fisheries of the River Axe - including the use of Conservation Limits as part of this process, (ii) to identify factors which may be limiting stock and fishery performance and (iii) to propose remedial measures address these factors. The salmon stock of the River Axe was apparently in a healthy state in the 1950s, supporting net and rod fisheries with average annual catches of around 100 and 50 fish respectively. Catches declined through the 1970s and 1980s to the extent where no salmon were recorded in the late 1980s and early 1990s. This decline was probably due largely to the effects of agricultural pollution, which virtually extinguished the salmon stock of the River Axe. Water quality has subsequently improved, but has deteriorated again in recent years.

The River Axe SAP Final Plan

This is the River Axe Salmon Action Plan Final document produced by the Environment Agency in 2004. The River Axe Salmon Action Plan (SAP) has been produced after consideration of feedback from public consultation. The final plan provides a list of the agreed issues and actions for the next five years to maintain and improve the salmon stock of the River Axe. Efforts have been made to identify possible sources of funding, partners and timescales. It indicates how the plan will be managed, including the process for reviewing stock status, issues, actions and progress. Low marine survival is currently a major factor limiting salmon stocks throughout the United Kingdom. However, on the River Axe the freshwater environment is still the main factor limiting the recovery of the salmon stock. Most of the adults returning to the Axe at present are probably derived from hatchery reared smolts, although there has been some natural reproduction in recent years. Juvenile populations in the Yarty indicate that this is a tributary where salmon have started to re-establish a self-sustaining population. Actions to improve the quality of the freshwater environment, both in terms of water quality and sedimentation, are seen as the top priorities, and are required to allow the Axe to support a self-sustaining salmon population.

Perfil dos HPA prioritários na exaustão de veículo a diesel, no combustível diesel utilizado durante os ensaios de emissão veicular e no óleo lubrificante do motor

As concentrações na exaustão e os fatores de emissão dos hidrocarbonetos policíclicos aromáticos (HPA) prioritários de um veículo a diesel e as suas respectivas concentrações no diesel usado durante os ensaios de emissão veicular foram determinados com a finalidade de estimar a contribuição dos HPA provenientes do combustível nas emissões. Os produtos da combustão foram coletados diretamente nas emissões brutas do escapamento, utilizando um sistema de amostragem a volume constante sem diluição dos gases da exaustão. Os HPA associados ao MP foram amostrados de forma estratificada, utilizando um impactador em cascata MOUDI e filtros de fibra de vidro como substratos, e os HPA em fase gasosa foram amostrados usando cartuchos de amberlite XAD-2. A concentração dos HPA no óleo lubrificante do motor também foi monitorada ao longo do tempo até a sua troca após 12.000 km de uso. Após a extração e tratamento das amostras, a identificação e quantificação dos HPA foram realizadas, utilizando cromatografia de fase gasosa acoplada à espectrometria de massas (CG-EM) com injetor de grande volume de vaporização com a temperatura programável (PTV-LVI). Cinco variáveis do PTV-LVI foram otimizadas, utilizando planejamento de experimentos, o que permitiu obter limites de detecção menores do que 2,0 g L-1. Somente 7 dos 16 HPA prioritários foram identificados na exaustão: NAP, ACY, ACE, FLU, FEN, FLT e PYR. Os ensaios de emissão veicular foram realizados com o veículo em modo estacionário, sem aplicação de carga e com baixa velocidade de rotação do motor (1500 rpm), utilizando um diesel com menor teor de enxofre (10 mg kg-1) e com 5% v/v de biodiesel. Esses fatores possivelmente contribuíram para reduzir as emissões dos outros 9 HPA a valores abaixo dos limites de detecção do método desenvolvido. Aproximadamente 80% da massa dos HPA totais associados ao MP estavam presentes em partículas com tamanho entre 1,0 m e 56 nm, e aproximadamente 4,5% estavam presentes em partículas menores do que 56 nm. Partículas menores que 2,5 m são facilmente inaladas e depositadas no trato respiratório e na região alveolar, justificando a preocupação com relação às emissões de HPA associados a partículas provenientes da exaustão veicular de motores a diesel. Somente 5 dos 7 HPA identificados na exaustão foram detectados no diesel: NAP, ACY, FLU, FEN e PYR. A razão entre os fatores de emissão (g L-1diesel) dos HPA na exaustão e suas respectivas concentrações do diesel (g L-1) variaram de 0,01 0,02 a 0,05 0,029, dependendo do HPA. Esses valores indicam que pelo menos 95 a 99% dos HPA identificados no diesel foram destruídos e/ou transformados em outros compostos durante a combustão, e/ou foram retidos no reservatório do óleo lubrificante. Por outro lado, os HPA que tiveram maiores concentrações no diesel também apresentaram maiores fatores de emissão, o que sugere que os HPA provenientes do diesel possuem uma contribuição significativa para as emissões dos HPA totais. O perfil dos HPA prioritários no óleo lubrificante mostrou-se semelhante ao perfil dos HPA no diesel e nas emissões totais, onde o NAP, FEN e PYR foram os HPA majoritários

Preparation of methyl tert-butyl ether (MTBE) over heteropoly acids immobilized on activated carbon (HPA/C) in the vapor phase

Heteropolyacids (HPAs) supported on the activated carbon (SiW12/C and PW12/C) have been used to study the formation of methyl tert-butyl ether (MTBE). Compared to the conventional commercial catalysts, Amberlyst-15 resin and HZSM-5, HPAs supported catalysts have been proved to have much higher catalytic activity under lower temperature, especially selectivity to MTBE is up to 100%. It may be due to the high acid strength of HPAs as well as the specialty of heteropolyanion.

Immobilization of the heteropoly acid (HPA) H4SiW12O40 (SiW12) on mesoporous molecular sieves (HMS and MCM-41) and their catalytic behavior

Supported catalysts, consisting of SiW12 immobilized on hexagonal mesoporous silica (HMS) and its aluminum-substituted derivative (MCM-41) with different loadings and calcination temperatures, have been prepared and characterized by X-ray diffraction, FT-IR and NH3-temperature programmed desorption. It is shown that SiW12 retains the Keggin structure on the mesoporous molecular sieves and no HPA crystal phase is developed, even at SiW12 loadings as high as 50 wt%. In the esterification of acetic acid by n-butanol, supported catalysts exhibit a higher catalytic activity and stability and held some promise of practical application. In addition, experimental results indicate that the loaded amount of SiW12 and the calcination temperatures have a significant influence on the catalytic activity, and the existence of aluminum has also an effect on the properties of supported catalysts.

气相酯化反应的高效催化剂──活性炭固载的杂多酸（HPA／C）

在气－固反应体系中研究了固载在活性炭上的硅钨（ＳｉＷ_（１２））和磷钨（ＰＷ_（１２））杂多酸在合成乙酸乙酯和乙酸丁酯中的催化性能。实验结果表明，反应温度、空间速度和反应物的配比均对催化活性有明显影响，并且，负载催化剂表面酸量与酯化反应活性之间存在着相应的顺变关系。催化剂在较低反应温度和相当大的流速下具有较高的酯化活性，且性能稳定，有一定的应用前景。

具有抗HIV病毒活性的杂多酸HPA－23与谷胱甘肽的作用

以谷胱甘肽作为病毒包络蛋白的模拟物，用ＮＭＲ方法研究了与具有抗爱滋病病毒活性杂多酸ＨＰＡ－２３的作用．对不同配比的ＨＰＡ－２３和谷胱甘肽混合物的１Ｈ和１８３ＷＮＭＲ谱研究结果表明，还原型和氧化型谷胱甘肽均以Ｃ末端ＣＯＯ－与ＨＰＡ－２３骨架的钨原子配位，还原型谷胱甘肽侧链上的巯基（ＳＨ）也参加配位．ＣＯＳＹ谱证明了ＳＨ配位为慢交换反应．早在七十年代初，人们就发现杂多酸具有抗病毒活性［１，２］．最近报道［ＮＨ４］１８［ＮａＳｂ９Ｗ２１Ｏ８８］·２４Ｈ２Ｏ（结构代号为ＨＰＡ－２３）具有很高的抗爱滋病病毒活性，在法国已进入临床应用［３］．但从分子水平研究杂多酸化合物抗病毒的机理．目前尚未见到国内外报道．而作为病毒可以广义地看作由一个蛋白外壳包裹，内部则为核酸．爱滋病病毒同样由两层蛋白所包裹，与宿主细胞发生吸附作用主要是通过外层包络蛋白（ＧＰ１２０）［４］，该蛋白的活性组份是一个由８个氨基酸组成的Ｔ（Ａｌａ－Ｓｅｒ－Ｔｈｒ－Ｔｈｒ－Ｔｈｒ－Ａｓｎ－Ｔｙｒ－Ｔｈｎ）肽段［５］．我们以容易得到的三肽一谷胱甘肽作为病毒包络蛋白的模拟物，用ＮＭＲ方法研究杂多酸ＨＰＡ－２３与它的作用．结果表明还原型和氧化型谷胱甘肽均以Ｃ末端Ｃ