Role of NOX enzymes in phagocyte response and signaling upon Chlamydia infection

Les membres de l'ordre des Chlamydiales peuvent infecter un choix étendu d'animaux, insectes, et protistes. Comme toutes bactéries intracellulaires obligatoires, les Chlamydiales ont besoin d'une cellule hôte pour se répliquer. Chaque fois qu'une cellule est infectée une lutte commence entre les mécanismes de défense de la cellule et l'arsenal de facteurs de virulence de la bactérie. Dans cette thèse nous nous sommes intéressés à déterminer le rôle de deux mécanismes de l'immunité innée de l'hôte. En premier, nous avons étudié les NADPH oxidases, une source de molécules superoxydantes (MSO). Leur rôle dans la restriction de la réplication de Waddlia chondrophila et Estrella iausannensis a été étudié dans l'organisme modèle Dictyostelium discoideum et les macrophages humains. Différentes protéines Nox étaient nécessaires pour contrôler la réplication de W. chondrophila ou E. Iausannensis. De plus, nous avons déterminé que parmi les Chlamydiales, cinq espèces possédaient une catalase. Cette enzyme peut dégrader l'eau oxygénée, une MSO. L'activité de la catalase a été démontrée in vitro et dans les corps élémentaires. Avant de pouvoir étudier le rôle de NOX2 dans des macrophages infectés avec E. Iausannensis, nous avons dû établir la capacité de la bactérie à se répliquer clans les macrophages avec son trafic intracellulaire. Le deuxième mécanisme d'immunité innée que nous avons étudié est l'autophagie. Dans les cellules infectées l'autophagie permet de digérer les bactéries envahissantes. Deux protéines de la voie autophagique (Atg1 et Atg8) jouent un rôle dans la restriction de la croissance de W. chondrophila dans D. discoideum. D'avantage d'études sur l'immunité innée et les bactéries apparentés aux Chlamydia sont indispensables, car les réponses paraissent être spécifiques pour chaque espèce. - Members of the Chlamydiales order are able to infect a large variety of animals, insects, and protists. These obligate intracellular bacteria require a host cell for replication. Each time a cell is infected a struggle begins between the virulence arsenal of the bacteria and the defense mechanisms activated by the host. Each bacterial species will exhibit a selection of virulence factors that will allow it to overcome the defense of the host in some species, but not others. In this thesis we were interested in dissecting the role of two host innate immunity mechanisms. First we determined the role of NADPH oxidases, a source of reactive oxygen species (ROS), in restricting replication of Waddlia chondrophila and EstreHa lausannensis in the model organism Dictyostelium discoideum and human macrophages. Different Nox proteins were required to restrict growth of W. chondrophila and E. lausannensis. Additionally, we determined that five Chlamydia- related bacterial species encode for catalase, an enzyme that is able to degrade hydrogen peroxide, a ROS. The activity of the catalase was demonstrated in vitro and in elementary bodies. To study the role of NOX2 in macrophages for E. lausannensis we first had to determine the ability of E. lausannensis to grow in macrophages. Besides demonstrating its replication we also determined the intracellular trafficking of E. lausannensis. The second innate immunity mechanism studied was autophagy. Through autophagy bacteria can be targeted to degradation. Atg1 and Atg8, two autophagic proteins appeared restrict W. chondrophila replication in D. discoideum. More studies on innate immunity and Chlamydia-related bacteria are required. It appears that the responses to innate immunity are species specific and it will be difficult to generalize data obtained for W. chondrophila to the Chlamydiales order.

Xanthine oxidase inhibition by febuxostat attenuates experimental atherosclerosis in mice.

Atherosclerosis is a chronic inflammatory disease due to lipid deposition in the arterial wall. Multiple mechanisms participate in the inflammatory process, including oxidative stress. Xanthine oxidase (XO) is a major source of reactive oxygen species (ROS) and has been linked to the pathogenesis of atherosclerosis, but the underlying mechanisms remain unclear. Here, we show enhanced XO expression in macrophages in the atherosclerotic plaque and in aortic endothelial cells in ApoE(-/-) mice, and that febuxostat, a highly potent XO inhibitor, suppressed plaque formation, reduced arterial ROS levels and improved endothelial dysfunction in ApoE(-/-) mice without affecting plasma cholesterol levels. In vitro, febuxostat inhibited cholesterol crystal-induced ROS formation and inflammatory cytokine release in murine macrophages. These results demonstrate that in the atherosclerotic plaque, XO-mediated ROS formation is pro-inflammatory and XO-inhibition by febuxostat is a potential therapy for atherosclerosis.

Anàlisi de la qualitat de l'aigua: estudi dels impactes humans sobre la qualitat de l'aigua en pous i basses del Parc del Garraf i rodalies

Amb la finalitat de conèixer l’estat de qualitat de les aigües de les basses i pous del Parc del Garraf, s’analitzen una sèrie de paràmetres fisico-químics en 17 estacions de mostreig prèviament seleccionades, distribuïdes en zones amb diferents tipologies d’ús del sòl. La base de l’anàlisi ha estat la integració d’informació provinent de diferents fonts. Mitjançant l’elaboració de taules i gràfics, la generació de cartografia i el tractament estadístic de les dades, s’ha procedit a la tria de punts de mostreig i s’ha obtingut un inventari que ha permès la interpretació global dels resultats, facilitant la diagnosi. El procés de tractament de dades inclou la confecció d’un índex de qualitat de les aigües (ICA) propi, no vinculant, a partir de fórmules genèriques de normalització i ponderació de valors. Durant la realització de la diagnosi s’han detectat pertorbacions puntuals en determinats paràmetres corresponents a contaminacions locals, en diferents estacions de mostreig. Aquestes pertorbacions s’han relacionat amb la situació dels pous i les basses al Garraf i les tipologies d’ús del sòl de cada zona. El diagnòstic de pertorbacions ha orientat les propostes de millora aplicables que s’han dividit en tres classes segons el nivell d’actuació. Aquestes incideixen principalment, en la millora de la informació disponible, l’aplicació de l’agricultura ecològica, l’explotació sostenible dels aqüífers i la realització d’estudis globals i/o locals, més complets i exhaustius.

Oxidative stress and inflammation response after nanoparticle exposure : differences between human lung cell monocultures and an advanced three-dimensional model of the human epithelial airways

Combustion-derived and manufactured nanoparticles (NPs) are known to provoke oxidative stress and inflammatory responses in human lung cells; therefore, they play an important role during the development of adverse health effects. As the lungs are composed of more than 40 different cell types, it is of particular interest to perform toxicological studies with co-cultures systems, rather than with monocultures of only one cell type, to gain a better understanding of complex cellular reactions upon exposure to toxic substances. Monocultures of A549 human epithelial lung cells, human monocyte-derived macrophages and monocyte-derived dendritic cells (MDDCs) as well as triple cell co-cultures consisting of all three cell types were exposed to combustion-derived NPs (diesel exhaust particles) and to manufactured NPs (titanium dioxide and single-walled carbon nanotubes). The penetration of particles into cells was analysed by transmission electron microscopy. The amount of intracellular reactive oxygen species (ROS), the total antioxidant capacity (TAC) and the production of tumour necrosis factor (TNF)-a and interleukin (IL)-8 were quantified. The results of the monocultures were summed with an adjustment for the number of each single cell type in the triple cell co-culture. All three particle types were found in all cell and culture types. The production of ROS was induced by all particle types in all cell cultures except in monocultures of MDDCs. The TAC and the (pro-)inflammatory reactions were not statistically significantly increased by particle exposure in any of the cell cultures. Interestingly, in the triple cell co-cultures, the TAC and IL-8 concentrations were lower and the TNF-a concentrations were higher than the expected values calculated from the monocultures. The interplay of different lung cell types seems to substantially modulate the oxidative stress and the inflammatory responses after NP exposure. [Authors]

Preserving Electronic Theses through the MetaArchive network

In the recent years most libraries have focused on mass digitization programs and keeping electronic born documents, showing and organizing them in a repository. While those repositories have evolved to a much more manageable systems focusing on the user expectations and introducing web 2.0 tools, digital preservation is still in the to-do list of most of them. There is quite a lot of studies focused on preservation and some complex models exist, unfortunately, very few practical systems are running and its quite difficult for a library to get involved in a solution already tested by others. The CBUC (Consortium of University Catalan Libraries) runs TDX, an ETD repository now keeping more than 10.000 full text thesis from any of the 12 university members. After 10 years running TDX a solid preservation system was needed to ensure every thesis would be kept as it was regardless what happens to the repository. The perfect solution was found in the MetaArchive cooperative, this is the effort of many insitutions to keep a copy of each other content through a newtwork using the LOCKSS software as a mechanism to keep track of any change. The presentation will shortly introduce what TDX and MetaArchive is but will, in a practical way, show how the LOCKSS network for presrervation works. Finally a summary of the benefits of the overall experience will be shown.

Nuevas tendencias en la construcción de la paz. Otra forma de innovación social

Este Working Paper tiene como objetivo identificar las nuevas formas de trabajo y tendencias emergentes en el campo de la construcción de la paz. El destinatario de este informe es el ICIP, por lo que las conclusiones están también enfocadas al plano operativo y a la posibilidad de integrar alguna de las prácticas descritas en su plan de actuación ordinaria. El texto presenta una serie de nuevas realidades emergentes, como el crowdfunding y el crowdsourcing, la transparencia financiera e informativa o el impacto de las redes sociales, que afectan a la forma en la que abordamos la construcción de la paz a nivel internacional. Cada tendencia viene acompañada de uno o varios ejemplos concretos que ayudan a comprender más fácilmente lo que está sucediendo en este terreno y qué podemos definir como innovación social.

LA DIRECCIÓ DE COMUNICACIÓ EN EL TERCER SECTOR

El Tercer Sector cívicosocial, entès com aquell conjunt d’organitzacions privades sense ànim de lucre que tenen com objectiu aconseguir la promoció de la persona, reduir les desigualtats socioeconòmiques i evitar l’exclusició social en el nostre territori, ha adquirit un pes molt significatiu en la nostra societat. És degut a aquesta importància, aconseguida al llarg dels anys, que totes les organitzaciones, fundacions i associacions han pres com a element de gran importància la comunicació. Saber comunicar-se ha arribat a un nivell imprescindible dins de la nostra societat i d’aquesta manera, el Tercer Sector n’ha pres consciència d’aquest fet. El present Treball Final de Carrera consisteix en l’estudi de diverses entitats representatives dels diversos sectors dins de l’àmbit territorial de Catalunya per veure de quina manera fan servir la comunicació i com es troben organitzades per saber assolir els seus objectius. Les nou entitats contactades són: Càritas, Intermón, Creu Roja, Amnistia Internacional, Greenpeace, Sos Racisme, SCIAS/ASC, Agrupació i ONCE. D’aquesta manera podem veure si el fet de disposar d’un departament dedicat a la comunicació, o de desenvolupar estratègies, repercuteix a obtenir uns millors resultats, tant interns com externs, dins de la entitat.

Anàlisi de la modelització hidrològica com a eina per la predicció d'inundacions i la gestió del territori

Les inundacions són actualment les catàstrofes naturals més recurrents i les que generen un major nombre de danys i víctimes arreu del món. L'ocupació de les zones inundables a les lleres del riu és la causa principal d’aquests desastres naturals. En aquest article es descriu la realització de models hidrològics com a mecanisme per la predicció d’inundacions i la gestió del territori. S’han estudiat les conques de la Riera de Santa Coloma (Catalunya) i del riu San Francisco (Guatemala) mitjançant els programes HEC-HMS i HEC-RAS, dels quals s’avalua la seva capacitat com eina per a la gestió del territori. S’ha analitzat l’efecte de la urbanització en el risc d’inundació en el cas de la Riera de Santa Coloma en base a la previsió del Plà d’Ordenament Urbanístic Municipal. S’han determinat les zones inundables resultants de episodis de precipitació extrems al Riu San Francisco per als episodis de les tempestes Stan(2005) i Agatha(2010).

MAP kinase pathways in UV-induced apoptosis of retinal pigment epithelium ARPE19 cells.

The retinal pigment epithelium (RPE) is constantly exposed to external injuries which lead to degeneration, dysfunction or loss of RPE cells. The balance between RPE cells death and proliferation may be responsible for several diseases of the underlying retina, including age-related macular degeneration (AMD) and proliferative vitreoretinopathy (PVR). Signaling pathways able to control cells proliferation or death usually involve the MAPK (mitogen-activated protein kinases) pathways, which modulate the activity of transcription factors by phosphorylation. UV exposure induces DNA breakdown and causes cellular damage through the production of reactive oxygen species (ROS) leading to programmed cell death. In this study, human retinal pigment epithelial cells ARPE19 were exposed to 100 J/m(2) of UV-C and MAPK pathways were studied. We first showed the expression of the three major MAPK pathways. Then we showed that activator protein-1 (AP-1) was activated through phosphorylation of cJun and cFos, induced by JNK and p38, respectively. Specific inhibitors of both kinases decreased their respective activities and phosphorylation of their nuclear targets (cJun and cFos) and reduced UV-induced cell death. The use of specific kinases inhibitors may provide excellent tools to prevent RPE apoptosis specifically in RPE diseases involving ROS and other stress-related compounds such as in AMD.

Las dos versiones de la Cuarta Sinfonía de Robert Schumann : primer caso de revisión de una sinfonía por parte del propio compositor

La Cuarta Sinfonía de Robert Schumann ha sido una obra clave del repertorio orquestal desde su triunfal estreno en 1853, considerándose una de las mejores creaciones del compositor. Sin embargo no es conocido el hecho de que esta sinfonía es en realidad la revisión de una obra de juventud que se estrenó sin éxito diez años antes, siendo Johannes Brahms quien, a finales del siglo XIX, defendió la mayor calidad de la Primera versión intentando revivirla, encontrando la oposición de Clara Schumann. Por lo tanto intentaré en este Proyecto Final estudiar en profundidad las dos versiones, analizando sus diferencias y planteando una hipótesis del por qué de la revisión del autor, presentando ambas versiones en el concierto en el que culmina este estudio.

Hepatitis C virus proteins promote mitochondrial bioenergetic dysfunction and nitro-oxidative stress: insights into pathogenesis

HCV-infection induces a state of oxidative stress more pronounced than in many other inflammatory diseases. Here we propose a temporal sequence of events in the HCV-infected cell whereby the primary alteration consists in release of Ca2+ from the ER followed by uptake into mitochondria. This triggers successive mitochondrial dysfunctions leading to generation of ROS and to a progressive metabolic adaptive response. Pathogenetic implications of the model and new opportunities for therapeutic intervention are discussed.

Novel loci for adiponectin levels and their influence on type 2 diabetes and metabolic traits: a multi-ethnic meta-analysis of 45,891 individuals.

Circulating levels of adiponectin, a hormone produced predominantly by adipocytes, are highly heritable and are inversely associated with type 2 diabetes mellitus (T2D) and other metabolic traits. We conducted a meta-analysis of genome-wide association studies in 39,883 individuals of European ancestry to identify genes associated with metabolic disease. We identified 8 novel loci associated with adiponectin levels and confirmed 2 previously reported loci (P = 4.5×10(-8)-1.2×10(-43)). Using a novel method to combine data across ethnicities (N = 4,232 African Americans, N = 1,776 Asians, and N = 29,347 Europeans), we identified two additional novel loci. Expression analyses of 436 human adipocyte samples revealed that mRNA levels of 18 genes at candidate regions were associated with adiponectin concentrations after accounting for multiple testing (p<3×10(-4)). We next developed a multi-SNP genotypic risk score to test the association of adiponectin decreasing risk alleles on metabolic traits and diseases using consortia-level meta-analytic data. This risk score was associated with increased risk of T2D (p = 4.3×10(-3), n = 22,044), increased triglycerides (p = 2.6×10(-14), n = 93,440), increased waist-to-hip ratio (p = 1.8×10(-5), n = 77,167), increased glucose two hours post oral glucose tolerance testing (p = 4.4×10(-3), n = 15,234), increased fasting insulin (p = 0.015, n = 48,238), but with lower in HDL-cholesterol concentrations (p = 4.5×10(-13), n = 96,748) and decreased BMI (p = 1.4×10(-4), n = 121,335). These findings identify novel genetic determinants of adiponectin levels, which, taken together, influence risk of T2D and markers of insulin resistance.

Colitis induced in mice with dextran sulfate sodium (DSS) is mediated by the NLRP3 inflammasome.

BACKGROUND: The proinflammatory cytokines interleukin 1beta (IL-1beta) and IL-18 are central players in the pathogenesis of inflammatory bowel disease (IBD). In response to a variety of microbial components and crystalline substances, both cytokines are processed via the caspase-1-activating multiprotein complex, the NLRP3 inflammasome. Here, the role of the NLRP3 inflammasome in experimental colitis induced by dextran sodium sulfate (DSS) was examined. METHODS: IL-1beta production in response to DSS was studied in macrophages of wild-type, caspase-1(-/-), NLRP3(-/-), ASC(-/-), cathepsin B(-/-) or cathepsin L(-/-) mice. Colitis was induced in C57BL/6 and NLRP3(-/-) mice by oral DSS administration. A clinical disease activity score was evaluated daily. Histological colitis severity and expression of cytokines were determined in colonic tissue. RESULTS: Macrophages incubated with DSS in vitro secreted high levels of IL-1beta in a caspase-1-dependent manner. IL-1beta secretion was abrogated in macrophages lacking NLRP3, ASC or caspase-1, indicating that DSS activates caspase-1 via the NLRP3 inflammasome. Moreover, IL-1beta secretion was dependent on phagocytosis, lysosomal maturation, cathepsin B and L, and reactive oxygen species (ROS). After oral administration of DSS, NLRP3(-/-) mice developed a less severe colitis than wild-type mice and produced lower levels of proinflammatory cytokines in colonic tissue. Pharmacological inhibition of caspase-1 with pralnacasan achieved a level of mucosal protection comparable with NLRP3 deficiency. CONCLUSIONS: The NLRP3 inflammasome was identified as a critical mechanism of intestinal inflammation in the DSS colitis model. The NLRP3 inflammasome may serve as a potential target for the development of novel therapeutics for patients with IBD.