Identification de nouveaux partenaires protéiques des récepteurs couplés aux protéines G contrôlant leur transport du reticulum endoplasmique à la membrane plasmique

Les récepteurs couplés aux protéines G (RCPGs) forment la plus grande et la plus diversifiée des familles de protéines localisées à la surface cellulaire et responsables de la transmission de signaux à l’intérieur des cellules. D’intenses recherches effectuées au cours des trente dernières années ont mené à l’identification de dizaines de protéines interagissant avec les RCPGs et contrôlant la signalisation, la désensibilisation, l’internalisation et la dégradation de ces importantes cibles pharmacologiques. Contrairement aux processus régulant l’activité des récepteurs à partir de la membrane plasmique, les mécanismes moléculaires contrôlant la biosynthèse des RCPGs dans le reticulum endoplasmique (RE) et leur transport jusqu’à la surface cellulaire sont très peu caractérisés. Une meilleure compréhension de ces processus nécessite l’identification de la machinerie protéique responsable de la maturation des RCPGs. Un crible protéomique basé sur le transfert d’énergie de résonance de bioluminescence (BRET), qui permet la mesure d’interactions protéiques dans les cellules vivantes, a mené à l’identification de plusieurs nouvelles protéines localisées dans la voie de sécrétion et interagissant potentiellement avec les RCPGs. Ces protéines étant localisées dans les compartiments cellulaires (reticulum endoplasmique et appareil de Golgi) responsables de la synthèse, du repliement adéquat et du transport à la membrane plasmique des récepteurs, il est très probable qu’elles soient impliquées dans le contrôle de l’expression des RCPGs à la surface cellulaire. La caractérisation de l’homologue humain de cornichon 4 (CNIH4), un nouvel intéracteur des RCPGs identifié dans le crible, a démontré que cette protéine localisée dans les compartiments précoces de la voie de sécrétion (RE et ERGIC) interagit de façon sélective avec les RCPGs. De plus, la suppression de l’expression endogène de cette protéine préalablement non-caractérisée, diminue le transport à la membrane plasmique d’un récepteur, indiquant que CNIH4 influence positivement l’export des RCPGs du RE. Ceci est supporté par l’observation que la surexpression de CNIH4 à de faibles niveaux favorise la maturation d’un récepteur mutant normalement retenu dans le RE. Nous avons également pu démontrer que CNIH4 est associée à la protéine Sec23, une des composantes de l’enveloppe des vésicules COPII qui sont responsables du transport des protéines du RE vers le Golgi, suggérant que CNIH4 pourrait favoriser le recrutement des récepteurs dans ces vésicules. La surexpression de CNIH4 à de très hauts niveaux provoque également la rétention intracellulaire des récepteurs. Cet effet dominant négatif pourrait être causé par la titration d’un autre facteur d’export des RCPGs. Une deuxième étude a permis de révéler que la protéine transmembranaire 9 (TMEM9), un nouvel intéracteur des RCPGs également identifié dans le crible, interagit sélectivement avec les récepteurs et avec CNIH4. La surexpression de cette protéine aux fonctions précédemment inconnues, rétablit le transport normal d’un récepteur en présence de CNIH4 surexprimée. De plus, la co-expression de TMEM9 potentialise la capacité de CNIH4 à augmenter la maturation d’un récepteur mutant normalement retenu dans le RE, suggérant que ces deux protéines forment un complexe régulant la maturation des RCPGs. Au cours de cette thèse, de nouvelles protéines interagissant avec les RCPGs et contrôlant leur expression à la membrane plasmique ont donc été identifiées, permettant une meilleure compréhension des mécanismes régulant le transport des récepteurs du RE à la surface cellulaire.

Cerbère au temps des " bio-maîtres " : la biométrie, servante-maîtresse d'une nouvelle ère biopolitique ? Le cas du programme US-VISIT

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal.

Biomethanation of syngas: identification of metabolic pathways from CO in a natural anaerobic consortium

Au cours des dernières décennies, l’intérêt pour la gazéification de biomasses a considérablement augmenté, notamment en raison de la grande efficacité de recouvrement énergétique de ce procédé par rapport aux autres procédés de génération de bioénergies. Les composants majoritaires du gaz de synthèse, le monoxyde de carbone (CO) et l’hydrogène (H2) peuvent entre autres servir de substrats à divers microorganismes qui peuvent produire une variété de molécules chimiques d’intérêts, ou encore produire des biocarburants, particulièrement le méthane. Il est donc important d'étudier les consortiums méthanogènes naturels qui, en syntrophie, serait en mesure de convertir le gaz de synthèse en carburants utiles. Cette étude évalue principalement le potentiel de méthanisation du CO par un consortium microbien issu d’un réacteur de type UASB, ainsi que les voies métaboliques impliquées dans cette conversion en conditions mésophiles. Des tests d’activité ont donc été réalisés avec la boue anaérobie du réacteur sous différentes pressions partielles de CO variant de 0.1 à 1,65 atm (0.09 à 1.31 mmol CO/L), en présence ou absence de certains inhibiteurs métaboliques spécifiques. Dès le départ, la boue non acclimatée au CO présente une activité carboxidotrophique relativement intéressante et permet une croissance sur le CO. Les tests effectués avec de l’acide 2- bromoethanesulfonique (BES) ou avec de la vancomycine démontrent que le CO est majoritairement consommé par les bactéries acétogènes avant d’être converti en méthane par les méthanogènes acétotrophes. De plus, un plus grand potentiel de méthanisation a pu être atteint sous une atmosphère constituée uniquement de CO en acclimatant auparavant la boue. Cette adaptation est caractérisée par un changement dans la population microbienne désormais dominée par les méthanogènes hydrogénotrophes. Ceci suggère un potentiel de production à large échelle de biométhane à partir du gaz de synthèse avec l’aide de biofilms anaérobies.

Identification et étude de mécanismes régulant l’expression de MAPK

Les fichiers accompagnant le document sont en format Microsoft Excel 2010.

Modifications post-traductionnelles des canaux calciques cardiaques de type L : identification des résidus asparagine qui participent à la glycosylation de la sous-unité auxiliaire CaVα2δ1

Les canaux calciques de type L CaV1.2 sont principalement responsables de l’entrée des ions calcium pendant la phase plateau du potentiel d’action des cardiomyocytes ventriculaires. Cet influx calcique est requis pour initier la contraction du muscle cardiaque. Le canal CaV1.2 est un complexe oligomérique qui est composé de la sous-unité principale CaVα1 et des sous-unités auxiliaires CaVβ et CaVα2δ1. CaVβ joue un rôle déterminant dans l’adressage membranaire de la sous-unité CaVα1. CaVα2δ1 stabilise l’état ouvert du canal mais le mécanisme moléculaire responsable de cette modulation n’a pas été encore identifié. Nous avons récemment montré que cette modulation requiert une expression membranaire significative de CaVα2δ1 (Bourdin et al. 2015). CaVα2δ1 est une glycoprotéine qui possède 16 sites potentiels de glycosylation de type N. Nous avons donc évalué le rôle de la glycosylation de type-N dans l’adressage membranaire et la stabilité de CaVα2δ1. Nous avons d’abord confirmé que la protéine CaVα2δ1 recombinante, telle la protéine endogène, est significativement glycosylée puisque le traitement à la PNGase F se traduit par une diminution de 50 kDa de sa masse moléculaire, ce qui est compatible avec la présence de 16 sites Asn. Il s’est avéré par ailleurs que la mutation simultanée de 6/16 sites (6xNQ) est suffisante pour 1) réduire significativement la densité de surface de! CaVα2δ1 telle que mesurée par cytométrie en flux et par imagerie confocale 2) accélérer les cinétiques de dégradation telle qu’estimée après arrêt de la synthèse protéique et 3) diminuer la modulation fonctionnelle des courants générés par CaV1.2 telle qu’évaluée par la méthode du « patch-clamp ». Les effets les plus importants ont toutefois été obtenus avec les mutants N663Q, et les doubles mutants N348Q/N468Q, N348Q/N812Q, N468Q/N812Q. Ensemble, ces résultats montrent que Asn663 et à un moindre degré Asn348, Asn468 et Asn812 contribuent à la biogenèse et la stabilité de CaVα2δ1 et confirment que la glycosylation de type N de CaVα2δ1 est nécessaire à la fonction du canal calcique cardiaque de type L.

La puissance technique de la médecine : des pistes de réflexion

Compte-rendu / Review

A Writer Identification System For Handwritten Malayalam Documents

Handwriting is an acquired tool used for communication of one's observations or feelings. Factors that inuence a person's handwriting not only dependent on the individual's bio-mechanical constraints, handwriting education received, writing instrument, type of paper, background, but also factors like stress, motivation and the purpose of the handwriting. Despite the high variation in a person's handwriting, recent results from different writer identification studies have shown that it possesses sufficient individual traits to be used as an identification method. Handwriting as a behavioral biometric has had the interest of researchers for a long time. But recently it has been enjoying new interest due to an increased need and effort to deal with problems ranging from white-collar crime to terrorist threats. The identification of the writer based on a piece of handwriting is a challenging task for pattern recognition. The main objective of this thesis is to develop a text independent writer identification system for Malayalam Handwriting. The study also extends to developing a framework for online character recognition of Grantha script and Malayalam characters

Identification of Putative Wintering Areas and Ecological Determinants of Population Dynamics of Common House-Martin (Delichon urbicum) and Common Swift (Apus apus) Breeding in Northern Italy

To identify the causes of population decline in migratory birds, researchers must determine the relative influence of environmental changes on population dynamics while the birds are on breeding grounds, wintering grounds, and en route between the two. This is problematic when the wintering areas of specific populations are unknown. Here, we first identified the putative wintering areas of Common House-Martin (Delichon urbicum) and Common Swift (Apus apus) populations breeding in northern Italy as those areas, within the wintering ranges of these species, where the winter Normalized Difference Vegetation Index (NDVI), which may affect winter survival, best predicted annual variation in population indices observed in the breeding grounds in 1992–2009. In these analyses, we controlled for the potentially confounding effects of rainfall in the breeding grounds during the previous year, which may affect reproductive success; the North Atlantic Oscillation Index (NAO), which may account for climatic conditions faced by birds during migration; and the linear and squared term of year, which account for nonlinear population trends. The areas thus identified ranged from Guinea to Nigeria for the Common House-Martin, and were located in southern Ghana for the Common Swift. We then regressed annual population indices on mean NDVI values in the putative wintering areas and on the other variables, and used Bayesian model averaging (BMA) and hierarchical partitioning (HP) of variance to assess their relative contribution to population dynamics. We re-ran all the analyses using NDVI values at different spatial scales, and consistently found that our population of Common House-Martin was primarily affected by spring rainfall (43%–47.7% explained variance) and NDVI (24%–26.9%), while the Common Swift population was primarily affected by the NDVI (22.7%–34.8%). Although these results must be further validated, currently they are the only hypotheses about the wintering grounds of the Italian populations of these species, as no Common House-Martin and Common Swift ringed in Italy have been recovered in their wintering ranges.

Megnin re-analysed: the case of the newborn baby girl, Paris, 1878

This paper analyses acarological evidence from a 130-year-old forensic investigation. It was the first case in forensic acarology, i.e., the first case where mites provided substantial information to estimate the post-mortem interval (PMI). In 1878, the mites found in the mummified body of a newborn baby girl in Paris, France, were studied by acarologist and forensic entomologist Jean Pierre M,gnin. M,gnin estimated around 2.4 million mites in the skull and identified them as Tyroglyphus longior (Gervais), a junior synonym of Tyrophagus longior. He suggested that the arrival of these mites at the corpse would have occurred by phoresy on carrier insects, roughly 5 months before the autopsy. There is no doubt about the identification of the mites, M,gnin was a highly respected acarologist. However, two main factors affecting the biology of Tyrophagus mites were not included in the original analysis. First, M,gnin stated that the mites were phoretic. However, he probably did not have access to information about the natural history of the species, because as a rule Tyrophagus mites are non-phoretic. Considering the omnipresence of Tyrophagus mites in soil, most likely the mites will have arrived almost immediately after death. Second, temperature was not taken into account during the estimations of the mite population growth rate. The new analysis is based on current knowledge of Tyrophagus biology and includes temperature, estimated following a handful of weather reports of the years 1877 and 1878. The new projections indicate that non-phoretic mites may have colonised the body just after death and the colony would have built up over 8 months, contrary to the 5 months proposed by M,gnin. This new lapse of time agrees with the PMI proposed by Brouardel: on 15 January 1878 he postulated the death of the newborn to have occurred some 8 months before the autopsy.

Detection of unusual mutation within the VP1 region of different re-isolates of poliovirus Sabin vaccine

In the present study, a genomic analysis of full VP1 sequence region of 15 clinical re-isolates (14 healthy vaccinees and one bone marrow tumor patient) was conducted, aiming to the identification of mutations and to the assessment of their impact on virus fitness, providing also insights relevant with the natural evolution of Sabin strains. Clinical re-isolates were analyzed by RT-PCR, sequencing and computational analysis. Some re-isolates were characterized by an unusual mutational pattern in which non-synonymous mutations outnumbered the synonymous ones. Furthermore, the majority of amino-acid substitutions were located in the capsid exterior, specifically in N-Ags, near N-Ags and in the north rim of the canyon. Also mutations, which are well-known determinants of attenuation, were identified. The results of this study propose that some re-isolates are characterized by an evolutionary pattern in which non-synonymous mutations with a direct phenotypic impact on viral fitness are fixed in viral genomes, in spite of synonymous ones with no phenotypic impact on viral fitness. Results of the present retrospective characterization of Sabin clinical re-isolates, based on the full VP1 sequence, suggest that vaccine-derived viruses may make their way through narrow breaches and may evolve into transmissible pathogens even in adequately immunized populations. For this reason increased poliovirus laboratory surveillance should be permanent and full VP1 sequence analysis should be conducted even in isolates originating from healthy vaccinees.

O impacto da coqueluche no Estado do Rio de Janeiroestudo epidemiológico e percepção da Biossegurança: contribuindo para análise da re-emergência no Brasil

A coqueluche é uma doença respiratória, causada pela bactéria Bordetella pertussis. Atualmente, estima-se a ocorrência anual de 50 milhões de casos e mais de 300 mil mortes anuais em todo mundo. A transmissão ocorre, principalmente, pelo contato direto de uma pessoa doente com uma pessoa suscetível, através de gotículas de secreção da orofaringe eliminada por tosse ou espirro. O estudo realizado objetivou a caracterização da coqueluche como doença re-emergente, visando a análise epidemiológica da doença no Estado do Rio de Janeiro, valorizando também a percepção da Biossegurança pelos profissionais da área da saúde. Os resultados alcançados revelaram indicadores da ressurgência da doença no Brasil. As análises foram objeto de reflexões propostas em quatro artigos científicos, que explicitaram as metodologias utilizadas, os resultados encontrados e as discussões pertinentes à pesquisa. Os artigos intitulam-se: (1) An overview of reemerging Pertussis and evidence of ressurgence in Brazil, (2); A re-emergência da coqueluche; Da rotina dos atendimentos ao imperativo da Biossegurança (3); Fórum itinerante de ciência e saúde. Programa de capacitação para as doenças negligenciadas e re-emergentes e (4) Identification of linear B epitopes of pertactin of Bordetella pertussis induced by immunization with whole and acellular vaccine

Are greenhouse gas signals of Northern Hemisphere winter extra-tropical cyclone activity dependent on the identification and tracking algorithm?

For Northern Hemisphere extra-tropical cyclone activity, the dependency of a potential anthropogenic climate change signal on the identification method applied is analysed. This study investigates the impact of the used algorithm on the changing signal, not the robustness of the climate change signal itself. Using one single transient AOGCM simulation as standard input for eleven state-of-the-art identification methods, the patterns of model simulated present day climatologies are found to be close to those computed from re-analysis, independent of the method applied. Although differences in the total number of cyclones identified exist, the climate change signals (IPCC SRES A1B) in the model run considered are largely similar between methods for all cyclones. Taking into account all tracks, decreasing numbers are found in the Mediterranean, the Arctic in the Barents and Greenland Seas, the mid-latitude Pacific and North America. Changing patterns are even more similar, if only the most severe systems are considered: the methods reveal a coherent statistically significant increase in frequency over the eastern North Atlantic and North Pacific. We found that the differences between the methods considered are largely due to the different role of weaker systems in the specific methods.

Contributions to the taxonomy of the Normanellidae (Copepoda, Harpacticoida): description of a new genus from the Brazilian continental shelf and re-assignment of Pseudocletodes vararensis Scott & Scott, 1893 (ex Nannopodidae)

A new genus and species of Normanellidae (Copepoda, Harpacticoida), Paranaiara inajae gen. et sp. nov., is described from the continental shelf off the northern coast of Sao Paulo State, Brazil. The new genus differs from the type genus Normanella Brady, 1880 and Sagamiella Lee & Huys, 1999 in its presence of lamelliform caudal rami, a maxillulary endopod represented by 2 setae, an unarmed maxillipedal syncoxa, and reduced setation on P2 enp-2 (without outer spine) and P3 enp-2 (with only 2 inner setae). All these apomorphic character states are shared with the genus Pseudocletodes Scott & Scott, 1893, formerly placed in the family Nannopodidae (ex Huntemanniidae) and here assigned to the Normanellidae. Pseudocletodes can be differentiated from Paranaiara by the loss of the P1 endopod and of the inner seta on P2-P4 enp-1, the presence of only 2 inner setae on P2 enp-2 (instead of 3) and only 1 inner seta on P4 exp-3 (instead of 2), the presence of a second inner seta on P4 enp-2 (instead of 1), the morphology of the fifth pair of legs which are not medially fused and have only 3 endopodal elements (instead of 4) in the male, and the well developed caudal ramus seta V (instead of rudimentary). It is postulated that prehensility of the P1 endopod was secondarily lost in the common ancestor of Paranaiara and Pseudocletodes. An updated family diagnosis of the Normanellidae and a dichotomous identification key to the 22 currently valid species are presented.

Re-framing risk transfer to assess integrative partnership opportunities: the S.A.R. approach and the AIG/BID/CNO regional surety bond facility of 2007

Identificar, compartilhar e gerenciar os riscos de contratar são preocupações que impedem o estabelicmento e a administração das Parcerias Públicos Particulares (PPP). Porem, gerentes das entidades públicas, bancos de formento, construtoras e seguradoras pesquisam e utilizam muitas técnicas para enfrentar a avaliação e gerenciamento dos riscos. A transferência de risco é uma indicação dos chamados benefícios que são inspirados pelos PPP, contudo devido às realidades contratuais e conceptuais, a entidade de cede o risco (o partido público) permanece quase sempre como o portador final do risco. Conseqüentemente, o partido público retem um interesse de resistência na gerência total destes riscos cedidos. Esta dissertação explora alguns defeitos das aproximações comuns a conceituar a gestão de risco no contexto de um PPP. Focalizando os conceitos da interdependência e da reciprocidade e usando na decisão para transferir o risco do projeto, esta dissertação molda a decisão para transferir o risco nos termos das realidades interdependentes de relacionamentos sistemáticos, alargam os conceitos técnicos do risco e da avaliação de risco, considerando o uso reflexivo das diferenças na analise de um estudo de caso. O autor explora estes conceitos em uma análise da decisão de um gerente de risco da empresa de construção civil brasileira Construtora Norberto Odebrecht (ODB) para projetar uma facilidade inovadora da ligação de garantia com Inter-American Development Bank (BID) e uma seguradora, American International Group (AIG), um negócio que ganhe o reconhecimento Trade Finance Magazine’s 2007 deal of the year. O autor mostra que por compreender a transferência de risco nos termos abordados nesta dissertação, um atore que transfere o risco pode identificar e criar mais oportunidades de estabelecer relacionamentos em longo prazo, através dos processos que a literatura atual do PPP ainda não considere. Os resultados devem fornecer contribuições para a pesquisas sobre a transferência do risco do projeto, na cooperação entre organizações e na seleção do sócio do projeto do potencial.

Metodologia para re-sintonia de controladores PID industriais

The present work has as objective to present a method of project and implementation of controllers PID, based on industrial instrumentation. An automatic system of auto-tunning of controllers PID will be presented, for systems of first and second order. The software presented in this work is applied in controlled plants by PID controllers implemented in a CLP. Software is applied to make the auto-tunning of the parameters of controller PID of plants that need this tunning. Software presents two stages, the first one is the stage of identification of the system using the least square recursive algorithm and the second is the stage of project of the parameters of controller PID using the root locus algorithm. An important fact of this work is the use of industrial instrumentation for the accomplishment of the experiments. The experiments had been carried through in controlled real plants for controllers PID implemented in the CLP. Thus has not only one resulted obtained with theoreticians experiments made with computational programs, and yes resulted obtained of real systems. The experiments had shown good results gotten with developed software