Geometry, petrology and growth of a shallow crustal laccolith : the Torres del Paine Mafic Complex (Patagonia)

1. ABSTRACTS - RÉSUMÉSSCIENTIFIC ABSTRACT - ENGLISH VERSIONGeometry, petrology and growth of a shallow crustal laccolith: the Torres del Paine Mafi c Complex (Patagonia)The Torres del Paine intrusive complex (TPIC) is a composite mafic-granitic intrusion, ~70km2, belonging to a chain of isolated Miocene plutons in southern Patagonia. Their position is intermediate between the Mesozoic-Cenozoic calc-alkaline subduction related Patagonian batholith in the West and the late Cenozoic alkaline basaltic back-arc related plateau lavas in the East. The Torres del Paine complex formed during an important reconfiguration of the Patagonian geodynamic setting, with a migration of magmatism from the arc to the back-arc, possibly related to the Chile ridge subductionThe complex intruded the flysch of the Cretaceous Cerro Toro and Punta Barrosa Formations during the Miocene, creating a well-defined narrow contact aureole of 200-400 m width.In its eastern part, the Torres del Paine intrusive complex is a laccolith, composed of a succession of hornblende-gabbro to diorite sills at its base, with a total thickness of ~250m, showing brittle contacts with the overlying granitic sills, that form spectacular cliffs of more than 1000m. This laccolith is connected, in the western part, to its feeding system, with vertical alternating sheets of layered gabbronorite and Hbl-gabbro, surrounded and percolated by diorites. ID-TIMS U-Pb on zircons on feeder zone (FZ) gab- bros yield 12.593±0.009Ma and 12.587±0.009Ma, which is identifcal within error to the oldest granite dated so far by Michel et al. (2008). In contrast, the laccolith mafic complex is younger than than the youngest granite (12.50±0.02Ma), and has been emplaced from 12.472±0.009Ma to 12.431 ±0.006Ma, by under-accretion beneath the youngest granite at the interface with previously emplaced mafic sills.The gabbronorite crystallization sequence in the feeder zone is dominated by olivine, plagioclase, clinopyroxene and orthopyroxene, while amphibole forms late interstitial crystals. The crystallization sequence is identical in Hornblende-gabbro from the feeder zone, with higher modal hornblende. Gabbronorite and Hornblende-gabbro both display distinct Eu and Sr positive anomalies. In the laccolith, a lower Hornblende-gabbro crystallized in sills and evolved to a high alkali shoshonitic series. The Al203, Ti02, Na20, K20, Ba and Sr composition of these gabbros is highly variable and increases up to ~50wt% Si02. The lower hornblende-gabbro is characterized by kaersutite anhedral cores with inclusions of olivine, clino- and orthopyroxene and rare apatite and An70 plagioclase. Trace element modelling indicates that hornblende and clinopyroxene are in equilibrium with a liquid whose composition is similar to late basaltic trachyandesitic dikes that cut the complex. The matrix in the lower hornblende gabbro is composed of normally zoned oligoclase, Magnesio-hornblende, biotite, ilmenite and rare quartz and potassium feldspar. This assemblage crystallized in-situ from a Ba and Sr-depleted melts. In contrast, the upper Hbl-gabbro is high-K calc-alkaline. Poikilitic pargasite cores have inclusions of euhedral An70 plagioclase inclusions, and contain occasionally clinopyroxene, olivine and orthopyroxene. The matrix composition is identical to the lower hornblende-gabbro and similar to the diorite. Diorite bulk rock compositions show the same mineralogy but different modal proportions relative to hornblende-gabbrosThe Torres del Paine Intrusive Complex isotopic composition is 87Sr/86Sr=0.704, 143Nd/144Nd=0.5127, 206Pb/204Pb=18.70 and 207Pb/204Pb=15.65. Differentiated dioritic and granitic units may be linked to the gabbroic cumulates series, with 20-50% trapped interstitial melt, through fractionation of olivine-bearing gabbronorite or hornblende-gabbro fractionation The relative homogeneity of the isotopic compositions indicate that only small amounts of assimilation occurred. Two-pyroxenes thermometry, clinopyroxene barometry and amphibole-plagioclase thermometry was used to estimate pressure and temperature conditions. The early fractionation of ultramafic cumulates occurs at mid to lower crustal conditions, at temperatures exceeding 900°C. In contrast, the TPIC emplacement conditions have been estimated to ~0.7±0.5kbar and 790±60°C.Based on field and microtextural observations and geochemical modelling, fractionation of basaltic-trachyandesitic liquids at intermediate to lower crustal levels, has led to the formation of the Torres del Paine granites. Repetitive replenishment of basaltic trachy- andesitic liquid in crustal reservoirs led to mixed magmas that will ascend via the feeder zone, and crystallize into a laccolith, in the form of successive dioritic and gabbroic sills. Dynamic fractionation during emplacement concentrated hornblende rich cumulates in the center of individual sills. Variable degrees.of post-emplacement compaction led to the expulsion of felsic liquids that preferentially concentrated at the top of the sills. Incremental sills amalgamation of the entire Torres del Paine Intrusive Complex has lasted for ~160ka.RESUME SCIENTIFIQUE - VERSION FRANÇAISEGéométrie, pétrologie et croissance d'un laccolite peu profond : Le complexe ma- fique du Torres del Paine (Patagonie)Le Complexe Intrusif du Torres del Paine (CITP) est une intrusion bimodale, d'environ 70km2, appartenant à une chaîne de plutons Miocènes isolés, dans le sud de la Patago-nie. Leur position est intermédiaire entre le batholite patagonien calco-alcalin, à l'Ouest, mis en place au Mesozoïque-Cenozoïque dans un contexte de subduction, et les basal-tes andésitiques et trachybasaltes alcalins de plateau, plus jeune, à l'Est, lié à l'ouverture d'un arrière-arc.A son extrémité Est, le CITP est une succession de sills de gabbro à Hbl et de diorite, sur une épaisseur de ~250m, avec des évidences de mélange. Les contacts avec les sills de granite au-dessus, formant des parois de plus de 1000m, sont cassants. Ce laccolite est connecté, dans sa partie Ouest, à une zone d'alimentation, avec des intrusions sub-ver- ticales de gabbronorite litée et de gabbro à Hbl, en alternance. Celles-ci sont traversées et entourées par des diorites. Les zircons des gabbros de la zone d'alimentation, datés par ID-TIMS, ont cristallisés à 12.593±0.009Ma et 12.587±0.009Ma, ce qui correspond au plus vieux granite daté à ce jour par Michel et al. (2008). A l'inverse, les roches manques du laccolite se sont mises en place entre 12.472±0.009Ma et 12.431 ±0.006Ma, par sous-plaquage successifs à l'interface avec le granite le plus jeune daté à ce jour (12.50±0.02Ma).La séquence de cristallisation des gabbronorites est dominée par Ol, Plg, Cpx et Opx, alors que la Hbl est un cristal interstitiel. Elle est identique dans les gabbros à Hbl de la zone d'alimentation, avec ~30%vol de Hbl. Les gabbros de la zone d'alimentation montrent des anomalies positives en Eu et Sr distinctes. Dans le laccolite, le gabbro à Hbl inférieur évolue le long d'une série shoshonitique, riche en éléments incompatibles. Sa concentration en Al203, Ti02, Na20, K20, Ba et Sr est très variable et augmente rapide-ment jusqu'à ~50wt% Si02. Il est caractérisé par la présence de coeurs résorbés de kaer- sutite, entourés de Bt, et contenant des inclusions d'OI, Cpx et Opx, ou alors d'Ap et de rares Plg (An70). Hbl et Cpx ont cristallisés à partir d'un liquide de composition similaire aux dykes trachy-andesite basaltique du CITP. La matrice, cristallisée in-situ à partir d'un liquide pauvre en Ba et Sr, est composée d'oligoclase zoné de façon simple, de Mg-Hbl, Bt, llm ainsi que de rares Qtz et KF. Le gabbro à Hbl supérieur, quant à lui, appartient à une suite chimique calco-alcaline riche en K. Des coeurs poecilitiques de pargasite con-tiennent de nombreuses inclusions de Plg (An70) automorphe, ainsi que des Ol, Cpx et Opx. La composition de la matrice est identique à celle des gabbros à Hbl inférieurs et toutes deux sont similaires à la minéralogie des diorites. Les analyses sur roches totales de diorites montrent la même variabilité que celles de gabbros à Hbl, mais avec une ten-eur en Si02 plus élevée.La composition isotopique des liquides primitifs du CITP a été mesurée à 87Sr/86Sr=0.704, 143Nd/144Nd=0.5127, 206Pb/204Pb=18.70 et 207Pb/204Pb=15.65. Les granites et diorites différenciés peuvent être reliés à des cumulais gabbronoritiques (F=0.74 pour les granites et F=1-0.5 pour les diorites) et gabbroïques à Hbl (fractionnement supplémentaire pour les granites, avec F=0.3). La cristallisation de 20 à 50%vol de liquide interstitiel piégé dans les gabbros du CITP explique leur signature géochimique. Seules de faibles quantités de croûte continentale ont été assimilées. La température et la pression de fractionnement ont été estimées, sur la base des thermobaromètres Opx-Cpx, Hbl-Plg et Cpx, à plus de 900°C et une profondeur correspondant à la croûte inférieure-moyenne. A l'inverse, les conditions de cristallisation de la matrice des gabbros et diorites du laccolite ont été estimées à 790±60°C et ~0.7±0.5kbar.Je propose que les liquides felsiques du CITP se soient formés par cristallisation frac-tionnée en profondeur des assemblages minéralogiques observés dans les gabbros du CITP, à partir d'un liquide trachy-andesite basaltique. La percolation de magma dans les cristaux accumulés permet la remontée du mélange à travers la zone d'alimentation, vers le laccolite, où des sills se mettent en place successivement. L'amalgamation de sills dans le CITP a duré ~160ka.Le CITP s'est formé durant une reconfiguration importante du contexte géodynamique en Patagonie, avec un changement du magmatisme d'arc vers un volcanisme d'arrière- arc. Ce changement est certainement lié à la subduction de la ride du Chili.RESUME GRAND PUBLIC - VERSION FRANÇAISEGéométrie, pétrologie et croissance d'une chambre magmatique peu profonde : Le complexe mafique du Torres del Paine (Patagonie)Le pourtour de l'Océan Pacifique est caractérisé par une zone de convergence de plaques tectoniques, appelée zone de subduction, avec le plongement de croûte océa-nique sous les Andes dans le cas de la Patagonie. De nombreux volcans y sont associés, formant la ceinture de feu. Mais seuls quelques pourcents de tout le magma traversant la croûte terrestre parviennent à la surface et la majeure partie cristallise en profondeur, dans des chambres magmatiques. Quelles est leur forme, croissance, cristallisation et durée de vie ? Le complexe magmatique du Torres del Paine représente l'un des meilleurs endroits au monde pour répondre à ces questions. Il se situe au sud de la Patagonie, formant un massif de 70km2. Des réponses peuvent être trouvées à différentes échelles, variant de la montagne à des minéraux de quelques 1000ème de millimètres.Il est possible de distinguer trois types de roches : des gabbros et des diorites sur une épaisseur de 250m, surmontées par des parois de granite de plus de 1000m. Les contacts entre ces roches sont tous horizontaux. Entre granites et gabbro-diorite, le contact est net, indiquant que le second magma s'est mis en place au contact avec un magma plus ancien, totalement solidifié. Entre gabbros et diorites, les contacts sont diffus, souvent non-linéaires, indiquant à l'inverse la mise en contact de magmas encore partiellement liquides. Dans la partie Ouest de cette chambre magmatique, les contacts entre roches sont verticaux. Il s'agit certainement du lieu de remplissage de la chambre magmatique.Lors du refroidissement d'un magma, différents cristaux vont se former. Leur stabilité et leur composition varient en fonction de la pression, de la température ou de la chimie du magma. La séquence de cristallisation peut être définie sur la base d'observations microscopiques et de la composition chimique des minéraux. Différents gabbros sont ainsi distingués : le gabbro à la base est riche en hornblende, d'une taille de ~5mm, sans inclusion de plagioclase mais avec des cristaux d'olivine, clinopyroxene et orthopyroxene inclus ; le gabbro supérieur est lui-aussi riche en hornblende (~5mm), avec les mêmes inclusions additionnées de plagioclase. Ces cristaux se sont formés à une température supérieure à 900°C et une profondeur correspondant à la croûte moyenne ou inférieure. Les minéraux plus fin, se trouvant hors des cristaux de hornblende des deux gabbros, sont similaires à ceux des diorites : plagioclase, biotite, hornblende, apatite, quartz et feldspath alcalin. Ces minéraux sont caractéristiques des granites. Ils ont cristallisé à ~790°C et ~2km de profondeur.La cristallisation des minéraux et leur extraction du magma par gravité provoque un changement progressif de la composition de ce dernier. Ainsi, après extraction d'olivine et d'orthopyroxene riches en Mg, de clinopyroxene riche en Ca, de plagioclase riche en Ca et Al et d'hornblende riche en Ca, Al et Mg, le liquide final sera appauvri en ces élé-ments. Un lien peut ainsi être proposé entre les diorites dont la composition est proche du liquide de départ, les granites dont la composition est similaire au liquide final, et les gabbros dont la minéralogie correspond aux minéraux extraits.L'utilisation de zircons, un minéral riche en U dont les atomes se transforment en Pb par décomposition radioactive au cours de millions d'années, permet de dater le refroidissement des roches qui les contiennent. Ainsi, il a été observé que les roches de la zone d'alimentation, à l'Ouest du complexe magmatique, ont cristallisés il y a 12.59±0.01 Ma, en même temps que les granites les plus vieux, se trouvant au sommet de la chambre magmatique, datés par Michel et al. (2008). Les deux roches pourraient donc avoir la même origine. A l'inverse, les gabbros et diorites de la chambre magmatique ont cristallisé entre 12.47±0.01Ma et 12.43±0.01Ma, les roches les plus vieilles étant à la base.En comparant la composition des roches du Torres del Paine avec celles d'autres en-tités géologiques de Patagonie, les causes du magmatisme peuvent être recherchées. A l'Ouest, on trouve en effet des intrusions granitiques, plus anciennes, caractéristiques de zones de convergence de plaque tectonique, alors qu'à l'Est, des laves basaltiques plus jeunes sont caractéristiques d'une dynamique d'extension. Sur la base des compositions chimiques des roches de ces différentes entités, l'évolution progressive de l'une à l'autre a pu être démontrée. Elle est certainement due à l'arrivée d'une dorsale océanique (zone d'extension crustale et de création de croûte océanique par la remontée de magma) dans la zone de subduction, le long des Andes.Je propose que, dans un premier temps, des magmas granitiques sont remontés dans la chambre magmatique, laissant d'importants volumes de cristaux dans la croûte pro-fonde. Dans un second épisode, les cristaux formés en profondeur ont été transportés à travers la croûte continentale, suite au mélange avec un nouveau magma injecté. Ces magmas chargés de cristaux ont traversé la zone d'alimentation avant de s'injecter dans la chambre magmatique. Différents puises ont été distingués, injectés dans la chambre magmatique du sommet à la base concernant les granites, puis à la base du granite le plus jeune pour les gabbros et diorites. Le complexe magmatique du Torres del Paine s'est construit sur une période totale de 160'000±20'000 ans.

In situ introducer sheath dilatation for complex aortic access.

Aortic access problems due to diseased or small peripheral vessels are a major issue in endovascular aneurysm repair (EVAR). In the emergency setting, like aortic rupture after blunt trauma, or in patients with a hostile abdomen, a more proximal access to the aorta is not a pleasant perspective. We developed in situ introducer sheath dilatation as a bail-out technique for patients with difficult aortic access under various circumstances including EVAR, intra-aortic balloon pump insertion and cannulation for perfusion. The method described allows to increase the access vessel diameter by 50% (from 6 to 9 mm) or the luminal circumference from 18 to 27 F. We have used this technique in five patients without complication, very much in contrast to the traditionally practiced 'forced device insertion'.

ARE PPARS NOVEL THERAPEUTIC TARGETS TO IMPROVE MELANOMA TREATMENT ?

Cutaneous melanoma is an aggressive malignant tumor of melanocytes, the pigment- producing cells of the epidermis, with a high incidence in developed countries. Despite some major clinical breakthroughs in the last few years, efficient therapies for metastatic melanoma, which portends a very bad prognosis, are still lacking. Among the potential therapeutic targets that have been attracting at-tention in melanoma are the peroxisome proliferator-activated receptors (PPARs). These members - a, ß and 7 - of the nuclear hormone receptor family, which are ligand-gated transcription factors endowed with a multitude of functions besides metabolism homeostasis, have displayed promising antitumor properties in a wide range of cancer cells, including melanoma. However, our knowledge of PPARs' functions in this skin cancer is far from complete, making the usefulness of any of the a, ß or 7 isotype as a therapeutic target uncertain. In this work, we showed that all three PPAR isotypes are expressed in normal melanocytes, in most melanoma cell lines and in primary and metastatic melanomas, and that PPAR/3 and 7 display transcriptional activity in normal melanocytes and melanoma cells. We also showed that the PPAR7 agonist rosiglitazone had anti-melanoma properties largely independent of PPAR7 expression, which was widely varying across the different cell lines and melanoma biopsies we evaluated and was not correlated with cell line stage. Consistent with the general view of PPAR7 as a tumor suppressor gene, we found that, in human samples, PPAR7 was less expressed in melanoma than in normal skin. Transcriptornic profiling of metastatic melanoma cells in which PPAR7 was pharmacologically modulated revealed an association with epithelial-to-mesenchymal transition, though the functional relevance of this finding remains to be determined. Collectively, our results suggests that PPAR7 activity in melanoma is highly complex and that a straightforward picture of PPAR7's role in this skin cancer is difficult to draw. In this study, we also provided compelling evidence that thioredoxin interacting protein (TXNIP) is, in melanoma, a bona fide PPAR7 target gene, the expression of which is repressed by PPAR7 activation. Although TXNIP is mostly known as an inhibitor of the major antioxidant thioredoxin, it has demonstrated a range of biological functions and is generally considered as a tumor suppressor gene. Consistently, we found that TXNIP expression is associated with growth arrest of melanoma cells in vitro and that forced expression of TXNIP strongly impairs cell proliferation. Interestingly, we also discovered that TXNIP favors melanoma cell migration while it diminishes their adhesion. Finally, we provided several lines of evidence that TXNIP may regulate these processes at the transcriptional level as well as by direct protein-protein interactions in the plasma membrane. Altogether, our findings suggest that the PPAR7 target TXNIP may be a double-edged sword in melanoma, hindering tumor growth but promoting invasion and dissemination. Experiments to evaluate the net biological outcome of TXNIP modulation in vivo are ongoing. -- Le mélanome cutané est une tumeur maligne agressive des mélanocytes, cellules de l'épiderme qui produisent la mélanine. Ce cancer présente un taux d'incidence élevé dans les pays développés et est grevé d'un pronostic très sombre une fois qu'il a disséminé. Malgré les importants progrès réalisés ces dernières années, aucune thérapie lie s'est encore montrée véritablement efficace contre le mélanome métastatique. Parmi les cibles thérapeutiques potentielles, nombre de groupes de recherche se sont penchés sur les peroxisome proliferator-activated receptors (PPARs). Ces récepteurs - a, ß et 7 - font partie de la famille des récepteurs nucléaires aux hormones, des facteurs de transcription activés par des ligands et dotés d'une multitude de fonctions en sus de la régulation du métabolisme. Ces protéines ont démontré des propriétés anti-tumorales prometteuses dans une large gamme de cellules cancéreuses, y compris le mélanome. Cependant, nous connaissons encore très mal les fonctions des PPARs dans ce cancer de la peau, rendant l'utilité thérapeutique de l'un des isotypes a, ß ou 7 incertaine. Dans ce travail, nous avons montré que les trois isotypes sont exprimés dans les mélanocytes normaux, dans la plupart des lignées de mélanome ainsi que dans des mélanomes primaires et métastatiques; nous avons aussi montré que PPAR/3 et 7 sont actifs sur le plan transcriptionnel dans les mélanocytes normaux et les cellules de mélanome. La rosiglitazone, un agoniste de PPAR7, a démontré des propriétés anti-mélanome essentiellement indépendantes de l'expression de PPAR7, qui semble très variable dans les lignées et les biopsies que nous avons évaluées; de plus, l'expression de PPAR7 n'est pas corrélée avec le stade de la lignée. En accord avec la vision communément admise de PPAR7 comme étant un gène suppresseur de tumeur, nous avons observé dans des échantillons humains que PPAR7 est moins exprimé dans les mélanomes que dans la peau normale. Une étude transcrip- tomique de cellules de mélanome métastatique a révélé que la modulation phar-macologique de PPAR7 est associée avec la transition épithélio-mésenchymateuse, même si la pertinence fonctionnelle de cette trouvaille reste à déterminer. Collec-tivement, ces résultats suggèrent que l'activité de PPAR/y dans le mélanome est hautement complexe et qu'une image claire du rôle de PPAR7 dans ce cancer est difficile à dessiner. Dans cette étude, nous avons également fourni de solides preuves que la thiore-doxin interacting protein (TXNIP) est, dans le mélanome, un gène cible bona fide de PPAR7 dont l'expression est réprimée par l'activation de PPAR7. Bien que TXNIP soit surtout connu comme un inhibiteur de la thiorédoxine -un anti-oxydant majeur - cette protéine a démontré une large gamme de fonctions biologiques et est généralement considérée comme un gène suppresseur de tumeur. En accord avec cette conception, nous avons trouvé que l'expression de TXNIP est associée avec l'arrêt de croissance des cellules de mélanome in vitro et que l'expression forcée de TXNIP freine considérablement la prolifération cellulaire. Nous avons aussi découvert que TXNIP favorise la migration des cellules de mélanome alors qu'elle diminue leur adhésion. Enfin, nous avons obtenu plusieurs preuves que TXNIP pourrait réguler ces processus tant au niveau transcriptionnel que par des interactions protéine-protéine au sein de la membrane plasmique. En conclusion, nos résultats suggèrent que la cible de PPAR7 TXNIP pourrait être une épée à double tranchant dans le mélanome, freinant la croissance tumorale mais favorisant l'invasion et la dissémination. Des expériences permettant d'évaluer l'effet biologique net de la modulation de TXNIP in vivo sont en cours.

Modeling complex wells with the multi-scale finite-volume method

In this paper, an extension of the multi-scale finite-volume (MSFV) method is devised, which allows to Simulate flow and transport in reservoirs with complex well configurations. The new framework fits nicely into the data Structure of the original MSFV method,and has the important property that large patches covering the whole well are not required. For each well. an additional degree of freedom is introduced. While the treatment of pressure-constraint wells is trivial (the well-bore reference pressure is explicitly specified), additional equations have to be solved to obtain the unknown well-bore pressure of rate-constraint wells. Numerical Simulations of test cases with multiple complex wells demonstrate the ability of the new algorithm to capture the interference between the various wells and the reservoir accurately. (c) 2008 Elsevier Inc. All rights reserved.

The alternatively spliced domain TnFnIII A1A2 of the extracellular matrix protein tenascin-C suppresses activation-induced T lymphocyte proliferation and cytokine production.

Several lines of evidences have suggested that T cell activation could be impaired in the tumor environment, a condition referred to as tumor-induced immunosuppression. We have previously shown that tenascin-C, an extracellular matrix protein highly expressed in the tumor stroma, inhibits T lymphocyte activation in vitro, raising the possibility that this molecule might contribute to tumor-induced immunosuppression in vivo. However, the region of the protein mediating this effect has remained elusive. Here we report the identification of the minimal region of tenascin-C that can inhibit T cell activation. Recombinant fragments corresponding to defined regions of the molecule were tested for their ability to inhibit in vitro activation of human peripheral blood T cells induced by anti-CD3 mAbs in combination with fibronectin or IL-2. A recombinant protein encompassing the alternatively spliced fibronectin type III domains of tenascin-C (TnFnIII A-D) vigorously inhibited both early and late lymphocyte activation events including activation-induced TCR/CD8 down-modulation, cytokine production, and DNA synthesis. In agreement with this, full length recombinant tenascin-C containing the alternatively spliced region suppressed T cell activation, whereas tenascin-C lacking this region did not. Using a series of smaller fragments and deletion mutants issued from this region, we have identified the TnFnIII A1A2 domain as the minimal region suppressing T cell activation. Single TnFnIII A1 or A2 domains were no longer inhibitory, while maximal inhibition required the presence of the TnFnIII A3 domain. Altogether, these data demonstrate that the TnFnIII A1A2 domain mediate the ability of tenascin-C to inhibit in vitro T cell activation and provide insights into the immunosuppressive activity of tenascin-C in vivo.

Transcriptional activation of the GLUT2 gene by the IPF-1/STF-1/IDX-1 homeobox factor.

The homeodomain protein PDX-1, referred as IPF-1/STF-1/IDX-1, is a transcriptional factor that plays a critical role in the control of several genes expressed in the pancreatic islet. PDX-1 gene expression has been previously shown to be reduced in cultured beta-cell lines chronically exposed to high glucose concentrations. As the glucose transporter type 2 (GLUT2) gene expression is selectively decreased in the beta-pancreatic cells of experimental models of diabetes, we postulated that the loss of GLUT2 gene expression in the pancreatic islets of diabetic animals may be due to the loss of PDX-1 transacting function on the GLUT2 gene. We, therefore, investigated the potential role of PDX-1 in the transcriptional control of GLUT2. We have identified a repeat of a TAAT motif (5'-TAATA-ATAACA-3') conserved in the sequence of the human and murine GLUT2 promoters. Recombinant PDX-1 binds to this GLUT2TAAT motif in electrophoretic mobility shift experiments. PDX-1 antiserum detects the formation of the complex of PDX-1 with the GLUT2TAAT motif in nuclear extracts from the pancreatic insulin-secreting cell line, beta TC3. The GLUT2TAAT motif was mutated in the murine GLUT2 promoter (-1308/+49 bp) linked to a luciferase reporter gene and transfected into beta TC3 cells. Compared with the transcriptional activity of the wild type promoter, that of the mutated promoter decreases by 41%. Multiple copies of the GLUT2TAAT motif were ligated 5' to a heterologous promoter and transfected into a PDX-1-expressing cell line (beta TC3) and into cell lines lacking the homeobox factor (InR1-G9 and JEG-3). The GLUT2TAAT motif mediates the activation of the heterologous promoter in the PDX-1-expressing cell line but not in InR1-G9 or JEG-3 cell lines. Furthermore, cotransfection in a PDX-1-deficient cell line with the expression vector encoding PDX-1 transactivates specifically the heterologous promoter containing the multimerized GLUT2TAAT motif. These data demonstrate that the murine GLUT2 promoter is controlled by the PDX-1 homeobox factor through the identified GLUT2TAAT motif.

Anopheles goeldii Rozeboom & Gabaldón (Diptera, Culicidae): a species of the Nuneztovari Complex of Anopheles Meigen

Anopheles (Nyssorhynchus) goeldiiRozeboom & Gabaldón, 1941, a species of the Nuneztovari Complex, was described based on morphological characteristics of the male, female, larva, pupa, and eggs. The type locality is Boa Vista (= Fordlândia), a district in the vicinity of Rio Tapajós, in the municipality of Aveiro, in the state of Pará, Brazil. Anopheles goeldii is redescribed based on morphological traits of the fourth instar larva, pupa, egg, and male and female. DNA sequences from the cytochrome oxidase subunit I (COI barcode region) of the mitochondrial genome were utilized for species characterization. Specimens of An. goeldii from the Pará, Amapá, and Amazonas states were employed to redescribe the species and to compare with morphologically similar taxa.

Photodamage to Oxygen Evolving Complex - An Initial Event in Photoinhibition of Photosystem II.

Photosystem II (PSII) of oxygenic photosynthesis is susceptible to photoinhibition. Photoinhibition is defined as light induced damage resulting in turnover of the D1 protein subunit of the reaction center of PSII. Both visible and ultraviolet (UV) light cause photoinhibition. Photoinhibition induced by UV light damages the oxygen evolving complex (OEC) via absorption of UV photons by the Mn ion(s) of OEC. Under visible light, most of the earlier hypotheses assume that photoinhibition occurs when the rate of photon absorption by PSII antenna exceeds the use of the absorbed energy in photosynthesis. However, photoinhibition occurs at all light intensities with the same efficiency per photon. The aim of my thesis work was to build a model of photoinhibition that fits the experimental features of photoinhibition. I studied the role of electron transfer reactions of PSII in photoinhibition and found that changing the electron transfer rate had only minor influence on photoinhibition if light intensity was kept constant. Furthermore, quenching of antenna excitations protected less efficiently than it would protect if antenna chlorophylls were the only photoreceptors of photoinhibition. To identify photoreceptors of photoinhibition, I measured the action spectrum of photoinhibition. The action spectrum showed resemblance to the absorption spectra of Mn model compounds suggesting that the Mn cluster of OEC acts as a photoreceptor of photoinhibition under visible light, too. The role of Mn in photoinhibition was further supported by experiments showing that during photoinhibition OEC is damaged before electron transfer activity at the acceptor side of PSII is lost. Mn enzymes were found to be photosensitive under visible and UV light indicating that Mn-containing compounds, including OEC, are capable of functioning as photosensitizers both in visible and UV light. The experimental results above led to the Mn hypothesis of the mechanism of continuous-light-induced photoinhibition. According to the Mn hypothesis, excitation of Mn of OEC results in inhibition of electron donation from OEC to the oxidized primary donor P680+ both under UV and visible light. P680 is oxidized by photons absorbed by chlorophyll, and if not reduced by OEC, P680+ may cause harmful oxidation of other PSII components. Photoinhibition was also induced with intense laser pulses and it was found that the photoinhibitory efficiency increased in proportion to the square of pulse intensity suggesting that laser-pulse-induced photoinhibition is a two-photon reaction. I further developed the Mn hypothesis suggesting that the initial event in photoinhibition under both continuous and pulsed light is the same: Mn excitation that leads to the inhibition of electron donation from OEC to P680+. Under laser-pulse-illumination, another Mn-mediated inhibitory photoreaction occurs within the duration of the same pulse, whereas under continuous light, secondary damage is chlorophyll mediated. A mathematical model based on the Mn hypothesis was found to explain photoinhibition under continuous light, under flash illumination and under the combination of these two.

Reconstruction fronto-orbitaire complexe avec prothèse en PEEK et expansion cutanée : à propos d'un cas [Complex fronto-orbital reconstruction with a PEEK prosthesis and skin expansion: about a case].

INTRODUCTION: Reconstructions of the fronto-orbital area remain a challenge to the reconstructive surgeon, due to the functional and esthetic impact. OBSERVATION: The authors present a case of a complex fronto-orbital reconstruction with a PEEK (PolyEtherEtherKetone) implant, associated with a skin expansion. DISCUSSION: With a follow-up of over three years, the cosmetic result is excellent. The authors believe that this technique is reliable, fast with long-term good results.

Isolated integrin beta3 subunit cytoplasmic domains require membrane anchorage and the NPXY motif to recruit to adhesion complexes but do not discriminate between beta1- and beta3-positive complexes.

Integrin adhesion receptors consist of non-covalently linked alpha and beta subunits each of which contains a large extracellular domain, a single transmembrane domain and a short cytoplasmic tail. Engaged integrins recruit to focal structures globally termed adhesion complexes. The cytoplasmic domain of the beta subunit is essential for this clustering. beta1 and beta3 integrins can recruit at distinct cellular locations (i.e. fibrillar adhesions vs focal adhesions, respectively) but it is not clear whether individual beta subunit cytoplasmic and transmembrane domains are by themselves sufficient to drive orthotopic targeting to the cognate adhesion complex. To address this question, we expressed full-length beta3 transmembrane anchored cytoplasmic domains and truncated beta3 cytoplasmic domains as GFP-fusion constructs and monitored their localization in endothelial cells. Membrane-anchored full-length beta3 cytoplasmic domain and a beta3 mutant lacking the NXXY motif recruited to adhesion complexes, while beta3 mutants lacking the NPXY and NXXY motifs or the transmembrane domain did not. Replacing the natural beta subunit transmembrane domain with an unrelated (i.e. HLA-A2 alpha chain) transmembrane domain significantly reduced recruitment to adhesion complexes. Transmembrane anchored beta3 and cytoplasmic domain constructs, however, recruited without discrimination to beta1- and beta3-rich adhesions complexes. These findings demonstrate that membrane anchorage and the NPXY (but not the NXXY) motif are necessary for beta3 cytoplasmic domain recruitment to adhesion complexes and that the natural transmembrane domain actively contributes to this recruitment. The beta3 transmembrane and cytoplasmic domains alone are insufficient for orthotopic recruitment to cognate adhesion complexes.

Recurrent deletions and reciprocal duplications of 10q11.21q11.23 including CHAT and SLC18A3 are likely mediated by complex low-copy repeats.

We report 24 unrelated individuals with deletions and 17 additional cases with duplications at 10q11.21q21.1 identified by chromosomal microarray analysis. The rearrangements range in size from 0.3 to 12 Mb. Nineteen of the deletions and eight duplications are flanked by large, directly oriented segmental duplications of >98% sequence identity, suggesting that nonallelic homologous recombination (NAHR) caused these genomic rearrangements. Nine individuals with deletions and five with duplications have additional copy number changes. Detailed clinical evaluation of 20 patients with deletions revealed variable clinical features, with developmental delay (DD) and/or intellectual disability (ID) as the only features common to a majority of individuals. We suggest that some of the other features present in more than one patient with deletion, including hypotonia, sleep apnea, chronic constipation, gastroesophageal and vesicoureteral refluxes, epilepsy, ataxia, dysphagia, nystagmus, and ptosis may result from deletion of the CHAT gene, encoding choline acetyltransferase, and the SLC18A3 gene, mapping in the first intron of CHAT and encoding vesicular acetylcholine transporter. The phenotypic diversity and presence of the deletion in apparently normal carrier parents suggest that subjects carrying 10q11.21q11.23 deletions may exhibit variable phenotypic expressivity and incomplete penetrance influenced by additional genetic and nongenetic modifiers.

FAM161A, associated with retinitis pigmentosa, is a component of the cilia-basal body complex and interacts with proteins involved in ciliopathies.

Retinitis pigmentosa (RP) is a retinal degenerative disease characterized by the progressive loss of photoreceptors. We have previously demonstrated that RP can be caused by recessive mutations in the human FAM161A gene, encoding a protein with unknown function that contains a conserved region shared only with a distant paralog, FAM161B. In this study, we show that FAM161A localizes at the base of the photoreceptor connecting cilium in human, mouse and rat. Furthermore, it is also present at the ciliary basal body in ciliated mammalian cells, both in native conditions and upon the expression of recombinant tagged proteins. Yeast two-hybrid analysis of binary interactions between FAM161A and an array of ciliary and ciliopathy-associated proteins reveals direct interaction with lebercilin, CEP290, OFD1 and SDCCAG8, all involved in hereditary retinal degeneration. These interactions are mediated by the C-terminal moiety of FAM161A, as demonstrated by pull-down experiments in cultured cell lines and in bovine retinal extracts. As other ciliary proteins, FAM161A can also interact with the microtubules and organize itself into microtubule-dependent intracellular networks. Moreover, small interfering RNA-mediated depletion of FAM161A transcripts in cultured cells causes the reduction in assembled primary cilia. Taken together, these data indicate that FAM161A-associated RP can be considered as a novel retinal ciliopathy and that its molecular pathogenesis may be related to other ciliopathies.

Dissociation of thymic positive and negative selection in transgenic mice expressing major histocompatibility complex class I molecules exclusively on thymic cortical epithelial cells.

Thymic positive and negative selection of developing T lymphocytes confronts us with a paradox: How can a T-cell antigen receptor (TCR)-major histocompatibility complex (MHC)/peptide interaction in the former process lead to transduction of signals allowing for cell survival and in the latter induce programmed cell death or a hyporesponsive state known as anergy? One of the hypotheses put forward states that the outcome of a TCR-MHC/peptide interaction depends on the cell type presenting the selecting ligand to the developing thymocyte. Here we describe the development and lack of self-tolerance of CD8(+) T lymphocytes in transgenic mice expressing MHC class I molecules in the thymus exclusively on cortical epithelial cells. Despite the absence of MHC class I expression on professional antigen-presenting cells, normal numbers of CD8(+) cells were observed in the periphery. Upon specific activation, transgenic CD8(+) T cells efficiently lysed syngeneic MHC class I(+) targets in vitro and in vivo, indicating that thymic cortical epithelium (in contrast to medullary epithelium and antigen-presenting cells of hematopoietic origin) is incapable of tolerance induction. Thus, compartmentalization of the antigen-presenting cells involved in thymic positive selection and tolerance induction can (at least in part) explain the positive/negative selection paradox.