INHIBITION OF THE MITOCHONDRIAL RESPIRATION OF TUMOR CELLS BY SOLUBLE FACTORS RELEASED BY ACTIVATED MACROPHAGES (CYTOTOXICITY, ELECTRON TRANSPORT, MONOCYTE)

Particular interest has been directed towards the macrophage as a primary antineoplastic cell due to its tumoricidal properties in vitro and the observation that an inverse relationship exists between the number of macrophages infiltrating a tumor and metastatic potential. The mechanism of macrophage-mediated injury of tumor cells remains unknown. Recently, it has been shown that injured tumor cells have defective mitochondrial respiration. Our studies have shown that activated macrophages can release soluble factors which can alter tumor cell respiration.^ The effects of a conditioned supernatant (CS) from cultures of activated macrophages on tumor cell (TC) mitochondrial respiration was studied. CS was obtained by incubation of BCG-elicited, murine peritoneal macrophage with RPMI-1640 supplemented with 10% FCS and 50 ng/ml bacterial endotoxin. This CS was used to treat cultures of EMT-6 TC for 24 hours. Mitochondrial respiration was measured polarigraphically using a Clark-type oxygen electrode. Cell growth rate was assessed by ('3)H-Thymidine incorporation. Exposure of EMT-6 TC to CS resulted in the inhibition of malate and succinate oxidation 76.6% and 72.9%, respectively. While cytochrome oxidase activity was decreased 61.1%. This inhibition was accompanied by a 98.8% inhibition of DNA synthesis (('3)H-Thymidine incorporation). Inhibition was dose-related with a 21.3% inhibition of succinate oxidase from a 0.3 ml dose of CS and a 50% inhibition with 1.0 mls. Chromatography of CS on Sephacryl S-200 resulted in isolation of an 80,000 and a 55,000 dalton component which contained the respiration inhibiting activity (RIF). These factors were distinct from a 120,000 dalton cytolytic factor determined by bioassay on Actinomycin-D treated L929 cells. RIF activity was also distinct from several other cytostatic factors but was itself associated with 2 peaks of cytostatic activity. Characterization of the RIF activity showed that it was destroyed by trypsin and heat (100(DEGREES)C, 5 min). It was stable over a broad range of pH (4-9) and its production was inhibited by cycloheximide. The RIF did not have a direct effect on isolated mitochondria of TC nor did it induce the formation of a stable intracellular toxin for mitochondria.^ In conclusion, activated macrophages synthesize and secrete an 80,000 and a 55,000 dalton protein which inhibits the mitochondrial metabolism of TC. These factors induce a cytostatic but not a cytolytic effect on TC.^ The macrophage plays a role in the control of normal and tumor cell growth and in tissue involution. Inhibition of respiration may be one mechanism used by macrophages to control cell growth.^

Ser88 phosphorylation regulates dynein light chain 1 (DLC1) function in the mouse mammary gland

Dynein light chain 1 (DLC1) is a highly conserved and ubiquitously expressed protein which might have critical cellular function as total loss of DLC1 caused Drosophila embryonic death. Despite many proteins and RNAs interaction with it identified, DLC1's function(s) and regulation are largely unknown. Recently, DLC1 was identified as a physiological substrate of P21-activate kinase 1(Pak1) kinase from a human mammary cDNA library in a yeast-2-hybridization screening assay. Studies in primary human tumors and cell culture implicated that DLC1 could promote mammary cancerous phenotypes, and more importantly, Ser88 phosphorylation of DLC1by Pak1 kinase was found to be essential for DLC1's tumorigenic activities. Based on the above tissue culture studies, we hypothesized that Ser88 phosphorylation regulates DLC1. ^ To test this hypothesis, we generated two transgenic mouse models: MMTV-DLC1 and MMTV-DLC1-S88A mice with mammary specific expression of the DLC1 and DLC1-S88A cDNAs. Both of the transgenic mice mammary glands showed rare tumor incidence which indicated DLC1 alone may not be sufficient for tumorigenesis in vivo. However, these mice showed a significant alteration of mammary development. Mammary glands from the MMTV-DLC1 mice had hyperbranching and alveolar hyperplasia, with elevated cell proliferation. Intriguingly, these phenotypes were not seen in the mammary glands from the MMTV-S88A mice. Furthermore, while MMTV-DLC1 glands were normal during involution, MMTV-S88A mice showed accelerated mammary involution with increase apoptosis and altered expression of involution-associated genes. Further analysis of the MMTV-S88A glands showed they had increased steady state level of Bim protein which might be responsible for the early involution. Finally, our in vitro data showed that Ser88 phosphorylation abolished DLC1 dimer and consequently might disturb its interaction with Bim and destabilize Bim. ^ Collectively, our findings provided in vivo evidence that Ser88 phosphorylation of DLC1 can regulate DLC1's function. In addition, Ser88 phosphorylation might be critical for DLC1 dimer-monomer transition. ^

Understanding the roles of Non-homologous end joining and p53 after DNA damage

The inability to maintain genomic stability and control proliferation are hallmarks of many cancers, which become exacerbated in the presence of unrepaired DNA damage. Such genotoxic stresses trigger the p53 tumor suppressor network to activate transient cell cycle arrest allowing for DNA repair; if the damage is excessive or irreparable, apoptosis or cellular senescence is triggered. One of the major DNA repair pathway that mends DNA double strand breaks is non-homologous end joining (NHEJ). Abrogating the NHEJ pathway leads to an accumulation of DNA damage in the lymphoid system that triggers p53-mediated apoptosis; complete deletion of p53 in this system leads to aggressive lymphomagenesis. Therefore, to study the effect of p53-dependent cell cycle arrest, we utilized a hypomorphic, separation-of-function mutant, p53p/p, which completely abrogates apoptosis yet retains partial cell cycle arrest ability. We crossed DNA ligase IV deficiency, a downstream ligase crucial in mending breaks during NHEJ, into the p53p/p background (Lig4-/-p53p/p). The accumulation of DNA damage activated the p53/p21 axis to trigger cellular senescence in developing lymphoid cells, which absolutely suppressed tumorigenesis. Interestingly, these mice progressively succumb to severe diabetes. Mechanistic analysis revealed that spontaneous DNA damage accumulated in the pancreatic b-cells, a unique subset of endocrine cells solely responsible for insulin production to regulate glucose homeostasis. The genesis of adult b-cells predominantly occurs through self-replication, therefore modulating cellular proliferation is an essential component for renewal. The progressive accumulation of DNA damage, caused by Lig4-/-, activated p53/p21-dependent cellular senescence in mutant pancreatic b-cells that lead to islet involution. Insulin levels subsequently decreased, deregulating glucose homeostasis driving overt diabetes. Our Lig4-/-p53p/p model aptly depicts the dichotomous role of cellular senescence—in the lymphoid system prevents tumorigenesis yet in the endocrine system leads to the decrease of insulin-producing cells causing diabetes. To further delineate the function of NHEJ in pancreatic b-cells, we analyzed mice deficient in another component of the NHEJ pathway, Ku70. Although most notable for its role in DNA damage recognition and repair within the NHEJ pathway, Ku70 has NHEJ-independent functions in telomere maintenance, apoptosis, and transcriptional regulation/repression. To our surprise, Ku70-/-p53p/p mutant mice displayed a stark increase in b-cell proliferation, resulting in islet expansion, heightened insulin levels and hypoglycemia. Augmented b-cell proliferation was accompanied with the stabilization of the canonical Wnt pathway, responsible for this phenotype. Interestingly, the progressive onset of cellular senescence prevented islet tumorigenesis. This study highlights Ku70 as an important modulator in not only maintaining genomic stability through NHEJ-dependent functions, but also reveals a novel NHEJ-independent function through regulation of pancreatic b-cell proliferation. Taken in aggregate, these studies underscore the importance for NHEJ to maintain genomic stability in b-cells as well as introduces a novel regulator for pancreatic b-cell proliferation.

Supplementarity measures on fuzzy sets.

In this paper, we commence the study of the so called supplementarity measures. They are introduced axiomatically and are then related to incompatibility measures by antonyms. To do this, we have to establish what we mean by antonymous measure. We then prove that, under certain conditions, supplementarity and incompatibility measuresare antonymous. Besides, with the aim of constructing antonymous measures, we introduce the concept of involution on the set made up of all the ordered pairs of fuzzy sets. Finally, we obtain some antonymous supplementarity measures from incompatibility measures by means of involutions.

Estrategias de desarrollo sostenible de la arquitectura del oasis de M’hamid, desierto del Sahara

Esta investigación se plantea con la hipótesis radical de cómo habitar el desierto de forma sostenible, desde una actitud pragmática y experimental basada en el progreso. La justificación se basa en primer lugar en los 2.000 millones de personas en el mundo que viven en entornos desérticos, el 80% de ellas, en países en desarrollo, porque el 40% de la superficie terrestre está bajo amenaza de desertificación afectando al 37% de la población mundial, con 12 millones de hectáreas al año perdidas por esa causa, y por último, porque se considera el desierto como un entorno de gran atractivo y potencial. El contenido de la investigación se estructura en tres movimientos: posicionamiento, mirada y acción: Desde el posicionamiento se define en primer lugar la sostenibilidad, aportando un nuevo diagrama donde se incorpora el ámbito arquitectónico como uno de los pilares principales, y, posteriormente, se establecen los criterios de evaluación de la sostenibilidad, aportando un sistema de indicadores donde se incorporan parámetros adecuados a las circunstancias del oasis. Del mismo modo, se estudian y analizan metodologías de actuación y proyectos de desarrollo sostenible existentes que enmarcan el estado del arte, constatando la dificultad de adaptación de los mismos a las condiciones de los oasis, por lo que se elabora una metodología propia donde se modifica la dinámica estratégica, de forma que el impulso se plantea desde la acción social, a través de hipótesis de estrategias basadas en sistemas low-cost, autoconstruidas, asumibles económicamente y de implantación factible. El caso de estudio específico radica en la situación extrema de las condiciones en el oasis de M’hamid, donde se evidencia un proceso de desintegración y abandono. Esto es debido a una acumulación de circunstancias externas e internas, de múltiples factores: naturales y antrópicos que afectan al oasis, llevando al extremo las condiciones climáticas y la escasez de recursos, naturales y artificiales. Factores como el cambio climático, la sequía, los cambios en las políticas del agua, la amenaza de desertificación, los conflictos sociales, el desequilibrio ecológico, la escasez económica, la crisis energética, la obsolescencia arquitectónica, el patrimonio construido prácticamente destruido, y la malentendida nueva arquitectura. Es importante constatar la escasa documentación gráfica existente sobre la zona de actuación lo que ha conllevado un amplio trabajo de documentación, tanto cartográfica como de observación directa, aportada a la tesis como investigación de elaboración propia. La mirada analítica al caso de estudio permite conocer los recursos disponibles y las potencialidades latentes del oasis de M’hamid, que permitirán actuar para subvertir la dinámica involutiva imperante, de forma que los dibujos iniciales de apropiación contextual y análisis críticos derivan en mapas de acción diagramados conformados por un sistema de objetos y la definición de estrategias transversales, deconstruyendo el pasado y reconstruyendo el futuro, incorporando sistemas alternativos que se definen en 7 líneas estratégicas de acción formuladas desde los 3 ámbitos relacionados con el ecosistema: ecológico, socio- económico y arquitectónico. Así, la tesis defiende la acción arquitectónica como impulsora del desarrollo sostenible, apoyada en 3 elementos: - la creación de objetos “tecnoartesanos”, para el aprovechamiento de los recursos energéticos - las transformaciones arquitectónicas, para reformular el hábitat desde la eficiencia energética y el progreso - y el impulso de acciones cotidianas, que redefinan las relaciones sociales, creando entornos cooperativos y colaborativos. En el ámbito ecológico se proponen actuaciones anti desertificación mediante incubadoras de árboles; sistemas alternativos de gestión del agua, como la lluvia sólida; estrategias de potenciación de la producción agrícola; la construcción de mecanismos de obtención de energía a partir de residuos, como los paneles solares con botellas PET. En el ámbito socioeconómico se plantean nuevas formas de acción social y de reactivación económica. Por último, en el ámbito urbano-arquitectónico, se incorporan modificaciones morfológicas a la arquitectura existente y una relectura contemporánea de la tierra, como material que permite nuevas geometrías, obteniendo arena petrificada por procesos microbiológicos, y potenciando la tierra como recurso artístico. Esta tesis es un punto de partida, recoge sistemas, estrategias y experiencias, para funcionar como un estímulo o impulso dinamizador del futuro desarrollo sostenible del oasis, abriendo vías de investigación y experimentación. ABSTRACT This research puts forth the radical hypothesis of how to inhabit the desert in a sustainable way, using a pragmatic and experimental approach based on progress. The justification for this resides in the fact that there are 2,000 million people in the world living in desert environments, 80% of them in developing countries. Forty percent of the earth’s surface is under threat of desertification, affecting 37% of the world population and with 12 million hectares being lost each year. And finally, the desert is considered as an attractive environment and therefore, with great potential. The content of the research is structured in three main sections: positioning, observation and action: As a point of departure, sustainability is defined, proposing a new framework where architecture is incorporated as one of the main pillars. Then, the criteria for evaluating sustainability are established. These provide a system of indicators, which incorporate parameters based on the specific circumstances of the oasis. Methodologies and existing sustainable development projects that represent the state-of-the-art are analyzed, discussing the difficulty of adapting them to conditions of oases. A methodology that modifies strategic concepts is developed, whereby the catalyst is social action, and strategies are developed based on low-cost, self-built, and feasible implementation systems. The specific case study lies in the extreme conditions in the oasis of M'hamid, where a process of decay and neglect is evident. This deterioration is due to an accumulation of external and internal circumstances, and of natural and anthropogenic factors that affect the oasis, leading to extreme weather conditions and a shortage of both natural and artificial resources. Factors include; climate change, drought, changes in water policies, the threat of desertification, social conflicts, ecological imbalance, economic shortage, the energy crisis, architectural obsolescence, destruction of built heritage, and misunderstood new architecture. It is important to note the extremely limited graphic information about the area has led me to produce an extensive archive of maps and drawings, many developed by direct observation, that contribute to the research. The case study analysis of the oasis of M'hamid examines the resources available and the latent potential to slow the prevailing trend towards deterioration. The initial drawings of contextual appropriation and critical analysis result in maps and diagrams of action, which are formed by a system of objects and the definition of strategies. These can be thought of as understanding or “deconstructing” the past to reconstruct the future. Alternative approaches defined in seven strategies for action are based on three fields related to the ecosystem: ecological, socioeconomic and architectural. Thus, the thesis defends architectural action to promote sustainable development, based on three elements: - The creation of "techno-artisans", to make use of energy resources - Architectural changes, to reformulate habitat in terms of energy efficiency and progress - And the promotion of everyday actions, to redefine social relations, creating cooperative and collaborative environments. In the ecological field, I propose anti-desertification actions such as; tree incubators, alternative water management systems(such as solid rain),; strategies to empower the agricultural production, energy from low-cost systems made out from recycled materials(such as solar panels from PET bottles or wind turbine from bicycle wheels). In the socioeconomic sphere, I propose to implement new forms of social action and economic regeneration. Finally, within the urban and architectural field, I propose morphological changes to the existing architecture and a contemporary reinterpretation of the earth as a material that allows new geometries, creating petrified sand by microbiological processes or enhancing nature as an artistic and energy resource. This thesis is a starting point. It collects systems, strategies and experiences to serve as a stimulus or dynamic momentum for future sustainable development of the oasis, opening new avenues of research and experimentation. RÉSUMÉ Cette recherche part d'une hypothèse radicale : comment habiter le désert de façon durable, et ce à partir d'une approche pragmatique et expérimentale basée sur le progrès. Cette hypothèse se justifie en raison des 2 milliards de personnes qui dans le monde habitent des environnements désertiques, 80% d'entre eux dans des pays en voie de développement, mais aussi parce que 40% de la surface de la planète est sous menace de désertification, un phénomène affectant 37% de la population mondiale et qui cause la perte de 12 millions d'hectares par an; et enfin parce que le désert est considéré comme un environnement très attrayant et fort d’un grand potentiel. Le contenu de la recherche se divise en trois mouvements: le positionnement, le regard et l'action : Du point de vue du positionnement on définit tout d'abord la durabilité, présentant un nouveau schéma où le domaine de l'architecture devient un des principaux piliers, et, par la suite, des critères d'évaluation de la durabilité sont établis, en fournissant un système d’indicateurs qui intègre les paramètres appropriés aux circonstances de l'oasis. De même, des méthodologies et des projets de développement durable existants sont étudiés et analysés, ce qui encadre l'état de l'art, remarquant la difficulté de les adapter aux conditions des oasis. De cette difficulté découle l'élaboration d'une méthodologie qui modifie la dynamique stratégique, de sorte que l'impulsion provient de l'action sociale, à travers des hypothèses de stratégie basées sur des systèmes low-cost, auto-construits, et de mise en oeuvre économiquement viable. Le cas d'étude spécifique réside en la situation extrême des conditions de l'oasis de M’hamid, où un processus de décadence et de négligence est évident. Cela est dû à une accumulation de circonstances externes et internes, de multiples facteurs: les facteurs naturels et anthropiques qui affectent l'oasis, menant à l'extrême les conditions météorologiques et la pénurie de ressources, autant naturelles qu'artificielles. Des facteurs tels que le changement climatique, la sécheresse, les changements dans les politiques de l'eau, la menace de la désertification, les conflits sociaux, le déséquilibre écologique, la pénurie économique, la crise de l'énergie, l'obsolescence architecturale, le patrimoine bâti pratiquement détruit et une mauvais compréhensif de la nouvelle architecture. Il est important de de faire remarquer le peu d'informations graphiques du domaine d'action, ce qui a conduit à un vaste travail de documentation, autant cartographique que relative à l'observation directe. Cette documentation s'ajoute à la thèse en tant que recherche propre. Le regard analytique sur le cas d'étude permet de connaître les ressources disponibles et le potentiel latent de l'oasis de M’hamid, qui agiront pour renverser la dynamique d'involution en vigueur. Ainsi, les premiers dessins d'appropriation contextuelle et analyse critique deviennent des cartes d'action schématisées formées par un système d'objets et la définition de stratégies transversales, qui déconstruisent le passé et reconstruisent l'avenir, en incorporant des systèmes alternatifs qui se définissent sur 7 lignes stratégiques d'action formulées à partir des 3 domaines en relation avec l’écosystème: l’écologique, le socio-économique et l'architectural. Ainsi, la thèse défend l'action architecturale en tant que promotrice du développement durable, et ce basé sur 3 éléments: - la création d'objets "technoartisans" pour l'exploitation des ressources énergétiques - les modifications architecturales, pour reformuler l'habitat du point de vue de l'efficacité énergétique et le progrès - et la promotion des actions quotidiennes, pour redéfinir les relations sociales, et la création d'environnements de coopération et collaboration. Dans le domaine de l'écologie des actions de lutte contre la désertification sont proposées à travers des pépinières d'arbres, des systèmes alternatifs de gestion de l'eau comme par exemple la pluie solide, des stratégies de mise en valeur de la production agricole, la construction de mécanismes de production d'énergie à partir de résidus, tels que les panneaux solaires ou les bouteilles en PET. Dans le domaine socio-économique, l'on propose de nouvelles formes d'action sociale et de reprise économique. Enfin, dans le domaine de l'urbain et de l'architectural, on incorpore des changements morphologiques à l'architecture existante et une relecture contemporaine de la terre, comme matériau qui permet de nouvelles géométries, en obtenant du sable pétrifié par des procédés microbiologiques et en mettant en valeur la terre comme une ressource artistique. Cette thèse n'est qu'un point de départ. Elle recueille des systèmes, des stratégies et des expériences pour servir de stimulus ou d'impulsion dynamisatrice du futur développement durable de l'oasis, en ouvrant des voies de recherche et d'expérimentation.

Dopamine β-hydroxylase deficiency impairs cellular immunity

Norepinephrine, released from sympathetic neurons, and epinephrine, released from the adrenal medulla, participate in a number of physiological processes including those that facilitate adaptation to stressful conditions. The thymus, spleen, and lymph nodes are richly innervated by the sympathetic nervous system, and catecholamines are thought to modulate the immune response. However, the importance of this modulatory role in vivo remains uncertain. We addressed this question genetically by using mice that lack dopamine β-hydroxylase (dbh−/− mice). dbh−/− mice cannot produce norepinephrine or epinephrine, but produce dopamine instead. When housed in specific pathogen-free conditions, dbh−/− mice had normal numbers of blood leukocytes, and normal T and B cell development and in vitro function. However, when challenged in vivo by infection with the intracellular pathogens Listeria monocytogenes or Mycobacterium tuberculosis, dbh−/− mice were more susceptible to infection, exhibited extreme thymic involution, and had impaired T cell function, including Th1 cytokine production. When immunized with trinitrophenyl-keyhole limpet hemocyanin, dbh−/− mice produced less Th1 cytokine-dependent-IgG2a antitrinitrophenyl antibody. These results indicate that physiological catecholamine production is not required for normal development of the immune system, but plays an important role in the modulation of T cell-mediated immunity to infection and immunization.

Thymus transplantation, a critical factor for correction of autoimmune disease in aging MRL/+mice.

MRL/MP-+/+ (MRL/+) mice develop pancreatitis and sialoadenitis after they reach 7 months of age. Conventional bone marrow transplantation has been found to be ineffective in the treatment of these forms of apparent autoimmune disease. Old MRL/+ mice show a dramatic thymic involution with age. Hematolymphoid reconstitution is incomplete when fetal liver cells (as a source of hemopoietic stem cells) plus fetal bone (FB; which is used to recruit stromal cells) are transplanted from immunologically normal C57BL/6 donor mice to MRL/+ female recipients. Embryonic thymus from allogeneic C57BL/6 donors was therefore engrafted along with either bone marrow or fetal hematopoietic cells (FHCs) plus fragments of adult or fetal bone. More than seventy percent of old MRL/+ mice (> 7 months) that had been given a fetal thymus (FT) transplant plus either bone marrow or FHCs and also bone fragments survived more than 100 days after treatment. The mice that received FHCs, FB, plus FT from allogeneic donors developed normal T cell and B cell functions. Serum amylase levels decreased in these mice whereas they increased in the mice that received FHCs and FB but not FT. The pancreatitis and sialoadenitis already present at the time of transplantations were fully corrected according to histological analysis by transplants of allogeneic FHCs, FB and FT in the MRL/+ mice. These findings are taken as an experimental indication that perhaps stem cell transplants along with FT grafts might represent a useful strategy for treatment of autoimmune diseases in aged humans.

O estudo das células Tγδ na manutenção da tolerância materna em vacas gestantes

O sistema imunológico materno desempenha um papel importante no estabelecimento da gestação e desenvolvimento de concepto até início do parto. A hipótese deste projeto é que há um recrutamento de células Tγδ para o endométrio ao longo da gestação que expressam citocinas que favorecem o estabelecimento e manutenção da tolerância materna a antígenos fetais durante a gestação em bovinos. Experimento I Para estudar a dinâmica populacional das células do sistema imune no sangue periférico de não lactantes não prenhes (NLNP), vacas lactantes no 1º trimestre e vacas no 3º trimestre da gestação, as PBMCs foram separadas por gradiente de densidade de Ficoll, seguido por protocolo de imunocitoquímica e analisadas em citômetro de fluxo para os anticorpos CD3+, CD4+, CD8+, CD14+, CD25+ e WC1+. As células analisadas tanto na região de linfócitos quanto na região de monócitos, não apresentaram diferença significativa entre os grupos analisados. Experimento II Para análise do perfil de expressão gênica das células Tγδ, foram coletadas amostras de sangue de vacas no 1º trimestre da gestação, vacas lactantes não prenhes (LNP) e vacas não lactantes não prenhes (NLNP). As células mononucleares foram separadas por gradiente de densidade de Ficoll e células Tγδ foram analisadas quanto ao perfil de citocinas por qRT-PCR para os genes IFNG; IL10; IL15; IL17; IL18; IL1B; IL4; IL-6; ISG15; PFR; TGFB2 e TNFA. A análise de expressão gênica mostrou tendência no aumento na expressão de IL1B, IL6 e TGFB2 em células PBMC em vacas NLNP quando comparado com vacas LNP e no 1º trimestre da gestação, enquanto que os outros genes analisados não apresentaram diferença significativa. Experimento III Fragmentos de endométrio, provenientes de abatedouro, foram coletados de vacas em 1º trimestre (33 a 35 dias de gestação), 2º trimestre (143 a 182 dias de gestação) e 3º trimestre de gestação (228 a 247 dias de gestação). Cortes congelados foram imunolocalizados e quantificados para as células do sistema imune CD3+, CD4+, CD8+, CD14+, CD18+, CD25+, CD62L+ e WC1+ por imunofluorescência. Nossos estudos mostraram aumento das células CD25+ e CD62L+ no endométrio no início da gestação. No meio da gestação, há um aumento das células WC1+ e CD14+. No final da gestação, observamos o aumento de CD14+, CD25+, CD18+ e CD62L+. Em suma, nossos dados sugerem que a modulação do sistema imune materno é específica para cada estágio da gestação, sendo que no início da gestação há um envolvimento de células T ativadas (CD25+) provavelmente para o estabelecimento de uma resposta ativa para tolerância dos antígenos fetais. Já no meio da gestação, há um recrutamento massivo de células Tγδ para o endométrio gravídico provavelmente para manter um microambiente de tolerância para o desenvolvimento fetal e no final da gestação células efetoras como macrófagos são recrutadas para o endométrio para auxiliar no processo do parto e involução uterina.

Human naïve regulatory T-cells feature high steady-state turnover and are maintained by IL-7

Naïve FoxP3-expressing regulatory T-cells (Tregs) are essential to control immune responses via continuous replenishment of the activated-Treg pool with thymus-committed suppressor cells. The mechanisms underlying naïve-Treg maintenance throughout life in face of the age-associated thymic involution remain unclear. We found that in adults thymectomized early in infancy the naïve-Treg pool is remarkably well preserved, in contrast to conventional naïve CD4 T-cells. Naïve-Tregs featured high levels of cycling and pro-survival markers, even in healthy individuals, and contrasted with other circulating naïve/memory CD4 T-cell subsets in terms of their strong γc-cytokine-dependent signaling, particularly in response to IL-7. Accordingly, ex-vivo stimulation of naïve-Tregs with IL-7 induced robust cytokine-dependent signaling, Bcl-2 expression, and phosphatidylinositol 3-kinase (PI3K)-dependent proliferation, whilst preserving naïve phenotype and suppressive capacity. Altogether, our data strongly implicate IL-7 in the thymus-independent long-term survival of functional naïve-Tregs, and highlight the potential of targeting the IL-7 pathway to modulate Tregs in different clinical settings.

Investigating the thymopoiesis stimulating property of interleukin-21 in aging mice

La vaccination est largement utilisée pour la génération de lymphocytes T spécifiques contre les tumeurs. Malheureusement, cette stratégie n'est pas adaptée aux personnes âgées car leur thymus régresse avec l'âge conduisant ainsi à une baisse dans la production de cellules T et à l'accumulation de cellules immunitaires âgées ayant des défauts liés à leurs stimulations. Comme il a été démontré auparavant que L’IL-21 est capable d’induire des fonctions thymiques, nous avons émis l’hypothèse que l’injection d’IL-21 à des souris âgées stimulera la thymopoïèse. Nos résultats montrent que l’administration de l’IL-21 augmente le nombre absolu de thymocytes chez les souris âgées et augmente la migration de ces cellules vers la périphérie ou ils contribuent à la diversité du TCR. De plus les cellules T en périphérie expriment un niveau plus élevé de miR181-a, et par conséquent moins de phosphatase comme SHP2, DUSP5/6 qui inhibent le TCR. En vaccinant des souris âgées avec le peptide Trp2, les souris traitées avec l’IL-21 montrent un retard dans la croissance des cellules B16 tumorales. Cette étude montre que l’IL-21 pourrait être utilisé comme stratégie pour le rétablissement du systeme immunitaire chez les personnes âgées.

Expression of the HPV16E7 oncoprotein by thymic epithelium is accompanied by disrupted T cell maturation and a failure of the thymus to involute with age

Transgenic mice expressing the E7 protein of HPV16 from the keratin 14 promoter demonstrate increasing thymic hypertrophy with age. This hypertrophy is associated with increased absolute numbers of all thymocyte types, and with increased cortical and medullary cellularity. In the thymic medulla, increased compartmentalization of the major thymic stromal cell types and expansion of thymic epithelial cell population is observed. Neither an increased rate of immature thymocyte division nor a decreased rate of immature thymocyte death was able to account for the observed hypertrophy. Thymocytes with reduced levels of expression of CD4 and/or CD8 were more abundant in transgenic (tg) mice and became increasingly more so with age. These thymic SP and DP populations with reduced levels of CD4 and/or CD8 markers had a lower rate of apoptosis in the tg than in the non-tg mice. The rate of export of mature thymocytes to peripheral lymphoid organs was less in tg animals relative to the pool of available mature cells, particularly for the increasingly abundant CD4lo population. We therefore suggest that mature thymocytes that would normally die in the thymus gradually accumulated in E7 transgenic animals, perhaps as a consequence of exposure to a hypertrophied E7-expressing thymic epithelium or to factors secreted by this expanded thymic stromal cell population. The K14E7 transgenic mouse thus provides a unique model to study effects of the thymic epithelial cell compartment on thymus development and involution.

Fibrés Vectoriels Semi-Stables sur une Courbe de Genre Deux et Association des Points dans L’espace Projectif

2000 Mathematics Subject Classification: 14D20, 14J60.

Potapov-Ginsburg Transformation and Functional Models of Non-Dissipative Operators

2000 Mathematics Subject Classification: Primary 47A20, 47A45; Secondary 47A48.

Calibrated embeddings in the special Lagrangian and coassociative cases

Every closed, oriented, real analytic Riemannian 3-manifold can be isometrically embedded as a special Lagrangian submanifold of a Calabi-Yau 3-fold, even as the real locus of an antiholomorphic, isometric involution. Every closed, oriented, real analytic Riemannian 4-manifold whose bundle of self-dual 2-forms is trivial can be isometrically embedded as a coassociative submanifold in a G_2-manifold, even as the fixed locus of an anti-G_2 involution. These results, when coupled with McLean's analysis of the moduli spaces of such calibrated submanifolds, yield a plentiful supply of examples of compact calibrated submanifolds with nontrivial deformation spaces.

British Romantic Criticism and the Fine Arts: A Study in Philosophical Theories of Literary Unity

This dissertation is an exploration of how a small but important group of Romantic critics, finding fault in the ideal of three unities developed by neoclassical Academicians and wrongly attributed to Aristotle, turned to the terminology and practices of the fine arts to emphasize their conception of organic unity in literature. The Romantic analogy to painting in particular enables a philosophical criticism of literature to present the aesthetic semblance of painting, the comprehension of a multitude of details in a harmonious whole that is a natural unity to its medium, as a paradigm of modern-romantic poetry and its aspirations to similar complexity, particularity, and imaginative colour. Further, in extension of the French Querelle des anciens et des modernes of the seventeenth century, the division of ancient and romantic art by Romantic critics like August Schlegel, Samuel Taylor Coleridge, and William Hazlitt not only establishes an ethnological and historical difference between the artistic productions of these two cultural periods but also allows, unlike the neoclassical unities, a non-anachronistic philosophical vocabulary of whole and parts or of the general and particular in the criticism of poetry, which involution provides a “rule” more consonant with the laws of the imagination rather than with the rhetorical and absolutist dicta that were thither available in the literary canon.