Mult-class differentiation of cannabis seedlings in a forensic context

This article presents an experimental study about the classification ability of several classifiers for multi-classclassification of cannabis seedlings. As the cultivation of drug type cannabis is forbidden in Switzerland lawenforcement authorities regularly ask forensic laboratories to determinate the chemotype of a seized cannabisplant and then to conclude if the plantation is legal or not. This classification is mainly performed when theplant is mature as required by the EU official protocol and then the classification of cannabis seedlings is a timeconsuming and costly procedure. A previous study made by the authors has investigated this problematic [1]and showed that it is possible to differentiate between drug type (illegal) and fibre type (legal) cannabis at anearly stage of growth using gas chromatography interfaced with mass spectrometry (GC-MS) based on therelative proportions of eight major leaf compounds. The aims of the present work are on one hand to continueformer work and to optimize the methodology for the discrimination of drug- and fibre type cannabisdeveloped in the previous study and on the other hand to investigate the possibility to predict illegal cannabisvarieties. Seven classifiers for differentiating between cannabis seedlings are evaluated in this paper, namelyLinear Discriminant Analysis (LDA), Partial Least Squares Discriminant Analysis (PLS-DA), Nearest NeighbourClassification (NNC), Learning Vector Quantization (LVQ), Radial Basis Function Support Vector Machines(RBF SVMs), Random Forest (RF) and Artificial Neural Networks (ANN). The performance of each method wasassessed using the same analytical dataset that consists of 861 samples split into drug- and fibre type cannabiswith drug type cannabis being made up of 12 varieties (i.e. 12 classes). The results show that linear classifiersare not able to manage the distribution of classes in which some overlap areas exist for both classificationproblems. Unlike linear classifiers, NNC and RBF SVMs best differentiate cannabis samples both for 2-class and12-class classifications with average classification results up to 99% and 98%, respectively. Furthermore, RBFSVMs correctly classified into drug type cannabis the independent validation set, which consists of cannabisplants coming from police seizures. In forensic case work this study shows that the discrimination betweencannabis samples at an early stage of growth is possible with fairly high classification performance fordiscriminating between cannabis chemotypes or between drug type cannabis varieties.

Plac8 is an inducer of C/EBPβ required for brown fat differentiation, thermoregulation, and control of body weight.

Brown adipocytes oxidize fatty acids to produce heat in response to cold or to excessive energy intake; stimulation of brown fat development and function may thus counteract obesity. Brown adipogenesis requires activation of the transcription factor C/EBPβ and recruitment of the zinc finger protein Prdm16, but upstream inducers of these proteins are incompletely defined. Here, we show that genetic inactivation of Plac8, a gene encoding an evolutionarily conserved protein, induces cold intolerance, and late-onset obesity, as well as abnormal morphology and impaired function of brown adipocytes. Using brown preadipocyte lines we show that Plac8 is required for brown fat differentiation, that its overexpression induces C/EBPβ and Prdm16, and that upon induction of differentiation Plac8 associates with C/EBPβ and binds to the C/EBPβ promoter to induce its transcription. Thus, Plac8 is a critical upstream regulator of brown fat differentiation and function that acts, at least in part, by inducing C/EBPβ expression.

Smooth muscle differentiation identifies two classes of poorly differentiated pleomorphic sarcomas with distinct outcome.

The clinical relevance of accurately diagnosing pleomorphic sarcomas has been shown, especially in cases of undifferentiated pleomorphic sarcomas with myogenic differentiation, which appear significantly more aggressive. To establish a new smooth muscle differentiation classification and to test its prognostic value, 412 sarcomas with complex genetics were examined by immunohistochemistry using four smooth muscle markers (calponin, h-caldesmon, transgelin and smooth muscle actin). Two tumor categories were first defined: tumors with positivity for all four markers and tumors with no or incomplete phenotypes. Multivariate analysis demonstrated that this classification method exhibited the strongest prognostic value compared with other prognostic factors, including histological classification. Secondly, incomplete or absent smooth muscle phenotype tumor group was then divided into subgroups by summing for each tumor the labeling intensities of all four markers for each tumors. A subgroup of tumors with an incomplete but strong smooth muscle differentiation phenotype presenting an intermediate metastatic risk was thus identified. Collectively, our results show that the smooth muscle differentiation classification method may be a useful diagnostic tool as well as a relevant prognostic tool for undifferentiated pleomorphic sarcomas.

The anti-apoptotic factor Bcl-2 can functionally substitute for the B cell survival but not for the marginal zone B cell differentiation activity of BAFF.

The TNF family ligand B cell-activating factor (BAFF, BLyS, TALL-1) is an essential factor for B cell development. BAFF binds to three receptors, BAFF-R, transmembrane activator and CAML interactor (TACI), and B cell maturation antigen (BCMA), but only BAFF-R is required for successful survival and maturation of splenic B cells. To test whether the effect of BAFF is due to the up-regulation of anti-apoptotic factors, TACI-Ig-transgenic mice, in which BAFF function is inhibited, were crossed with transgenic mice expressing FLICE-inhibitory protein (FLIP) or Bcl-2 in the B cell compartment. FLIP expression did not rescue B cells, while enforced Bcl-2 expression restored peripheral B cells and the ability to mount T-dependent antibody responses. However, many B cells retained immaturity markers and failed to express normal amounts of CD21. Marginal zone B cells were not restored and the T-independent IgG3, but not IgM, response was impaired in the TACI-IgxBcl-2 mice. These results suggest that BAFF is required not only to inhibit apoptosis of maturating B cells, but also to promote differentiation events, in particular those leading to the generation of marginal zone B cells.

Population differentiation and its consequences in the white campion "Silene latifolia" (Caryophyllaceae): a functional analysis

Summary In his theory On the Origin of Species by Means of Natural Selection (1859), Darwin describes evolution as a gradual change in population over time and that natural selection is a process that caused evolution. Because quantitative variation in species is partly influenced by several genes and thus heritable, association between levels of genetic variation at neutral markers and at quantitative traits and their partitioning within and among populations are important to study mechanisms that drive evolution in populations. Most studies addressing quantitative variation in plants focused on morphological and life history traits but not in traits affecting reproductive success. The aim of this thesis is to better understand how patterns of variation for neutral molecular markers and phenotypic traits drive the evolution of reproduction and defensive mechanisms in six European populations of Silene latifolia, a dioecious plant species. We found evidence for extremely high within and between population variation at six microsatellite loci and at most quantitative traits studied in plants grown under standardized conditions (morphology, life history and reproductive traits). Interestingly, there was clinal variation between age at first flowering and latitude. This pattern is likely due to natural selection since differentiation of this trait was high, heritable and probably higher than differentiation at neutral markers. Our study focused on sex specific selective pressures: mechanisms of intersexual coadaptation and defence mechanism against the seed predator Hadena bicruris. To address divergence at reproductive traits, we studied male and female population of origin effects and in particular pollen competitive ability on male post-pollination success in the study populations with within and between populations crosses. We crossed the same female plant with pollen from a male within the same population of origin and pollen from two males from two distinct populations, using a fixed tester male as a competitor. Additionally, we conducted control crosses with pollen from each male as a single donor. We analysed paternity success of each competitor with two microsatellite loci, seed set and offspring fitness. Male population of origin showed significant among-population variation for siring success at pollen competition. In vitro pollen germination rate showed heritable variation among populations and was positively correlated to siring success. Local or foreign pollen did not have a consistent advantage. Furthermore, female population of origin affected the outcome of pollen competition in some populations. There was no difference of seed set or offspring fitness in within/ between population crosses. This suggests that reproductive divergence may occur via pollen competition in Silene latifolia. The specialist seed predator Hadena bicruris may also induce divergence between populations. We tested potential constitutive and induced defence mechanisms against the specialist predator Hadena bicruris. Because fruit wall thickness is smaller in the invasive range (Northern America) were the moth is absent, this suggests that a thicker fruit wall is a potentially defensive trait against larval attack, and that relaxed selection in the absence of the seed predator has resulted in an evolutionary loss of this defence in the invasive range. Fruit wall thickness was different among three populations. Experimental exposure to moth eggs increased fruit abortion. Fruits built after attack on exposed plants did not have thicker fruit walls compared to fruits on non-exposed plants. Furthermore, fruits with thicker fruit walls were not less profitable, nor did they require longer handling time when exposed to larvae, suggesting no defensive role of fruit wall thickness. Our results show that there is high molecular and phenotypic variation in Silene latifolia and that traits potentially involved in reproductive success both for intra-specific (between sexes) and inter-specific interactions are heritable. Different selective forces may thus interact and cause differential evolution of geographically separated Silene latifolia populations in Europe, leading to the observed differentiation. Résumé Dans sa théorie de l'évolution, L'origine des espèces, ch. 4 (1859), Darwin décrit l'évolution comme un processus continu au cours du temps à l'intérieur de populations et que la sélection naturelle en est le moteur. La variation quantitative est en partie déterminée par plusieurs gènes, donc transmissible à la descendance. Associer le niveau de variation génétique à des marqueurs neutres au niveau de la variation à des traits quantitatifs, ainsi que la répartition à l'intérieur et entre les populations d'une espèce donnée de cette variation, sont importants dans la compréhension des forces évolutives. La plupart des études scientifiques sur la variation quantitative chez les plantes se sont intéressées à la morphologie et à la phénologie mais pas aux caractères impliqués dans le succès reproducteur. L'objectif de cette thèse est de mieux comprendre comment la répartition de la variation à des marqueurs neutres et des caractères quantitatifs influence l'évolution de la reproduction et des mécanismes de défense dans six populations Européennes de l'espèce dioïque Silene latifolia. Nous avons mis en évidence une grande diversité intra et inter-population à six loci microsatellites ainsi qu'à la plupart des caractères quantitatifs mesurés (morphologie, phénologie et traits reproducteurs) sur des plantes cultivées dans des conditions standardisées. Un résultat intéressant est la présence d'un cline latitudinal pour l'âge à la floraison. Ceci est probablement une conséquence de la sélection naturelle, puisque ce caractère est différencié entre les populations étudiées, héritable et que la différenciation de ce trait est supérieure à la différenciation des marqueurs neutres étudiés. Notre étude a ensuite porté plus précisément sur les pressions de sélection spécifiques aux sexes : la coadaptation entre les sexes et les mécanismes de défense contre l'insecte granivore Hadena bicruris. Afin d'évaluer la divergence sur les traits reproducteurs, nous avons étudié les effets des populations d'origine des mâles et des femelles et en particulier le succès reproducteur des mâles après pollinisation à l'aide de croisements inter et intra-population. Nous avons pollinisé la même femelle avec du pollen provenant d'un mâle de la même population ainsi qu'avec le pollen de deux mâles provenant de deux autres populations en situation de compétition avec un pollen provenant d'une population test. Des croisements contrôle ont été réalisés avec les mêmes mâles en pollinisation pure. Nous avons évalué le succès reproducteur de chaque mâle à l'aide d'analyses de paternité ainsi que la production de graines et la fitness de la descendance. L'origine du mâle avait un effet sur la paternité. Le taux de croissance in vitro du pollen est un caractère héritable et a eu un effet positif sur le succès reproducteur. De plus, l'origine de la femelle avait un effet sur le succès des mâles en compétition dans certaines populations. Nos résultats suggèrent qu'une divergence reproductive chez Silene latifolia pourrait apparaître suite à la compétition pollinique. Nous avons ensuite testé des mécanismes potentiels de défense constitutive et induite contre l'herbivore spécialiste Hadena bicruris, un papillon nocturne qui pourrait aussi jouer un rôle dans la différenciation des populations. L'épaisseur des fruits étant plus faible dans les régions où la plante est invasive (Amérique du Nord) et où l'insecte est absent, ce trait pourrait jouer un rôle défensif. Une pression de sélection plus faible causée par l'absence de l'herbivore aurait abouti à une perte de cette défense dans ces régions. Nous avons montré que l'épaisseur du fruit est variable selon les populations. L'infestation artificielle de fruit par l'insecte induit l'abscission sélective des fruits. Les fruits produits après une infestation n'étaient pas plus épais que les fruits issus de plantes non infestées. De plus, les fruits épais n'étaient pas moins nutritifs et ne causaient pas de perte de temps pour la prédation pour les larves, ce qui suggère que l'épaisseur des fruits ne joue pas un rôle défensif. Nos résultats montrent que plusieurs pressions de sélection interviennent et interagissent dans l'évolution de populations distantes, provoquant la divergence des populations Européennes de l'espèce Silene latifolia.

PPARbeta/delta regulates paneth cell differentiation via controlling the hedgehog signaling pathway.

BACKGROUND & AIMS: All 4 differentiated epithelial cell types found in the intestinal epithelium derive from the intestinal epithelial stem cells present in the crypt unit, in a process whose molecular clues are intensely scrutinized. Peroxisome proliferator-activated receptor beta (PPARbeta) is a nuclear hormone receptor activated by fatty acids and is highly expressed in the digestive tract. However, its function in intestinal epithelium homeostasis is understood poorly. METHODS: To assess the role of PPARbeta in the small intestinal epithelium, we combined various cellular and molecular approaches in wild-type and PPARbeta-mutant mice. RESULTS: We show that the expression of PPARbeta is particularly remarkable at the bottom of the crypt of the small intestine where Paneth cells reside. These cells, which have an important role in the innate immunity, are strikingly affected in PPARbeta-null mice. We then show that Indian hedgehog (Ihh) is a signal sent by mature Paneth cells to their precursors, negatively regulating their differentiation. Importantly, PPARbeta acts on Paneth cell homeostasis by down-regulating the expression of Ihh, an effect that can be mimicked by cyclopamine, a known inhibitor of the hedgehog signaling pathway. CONCLUSIONS: We unraveled the Ihh-dependent regulatory loop that controls mature Paneth cell homeostasis and its modulation by PPARbeta. PPARbeta currently is being assessed as a drug target for metabolic diseases; these results reveal some important clues with respect to the signals controlling epithelial cell fate in the small intestine.

Simultaneous coexpression of memory-related and effector-related genes by individual human CD8 T cells depends on antigen specificity and differentiation.

Phenotypic and functional cell properties are usually analyzed at the level of defined cell populations but not single cells. Yet, large differences between individual cells may have important functional consequences. It is likely that T-cell-mediated immunity depends on the polyfunctionality of individual T cells, rather than the sum of functions of responding T-cell subpopulations. We performed highly sensitive single-cell gene expression profiling, allowing the direct ex vivo characterization of individual virus-specific and tumor-specific T cells from healthy donors and melanoma patients. We have previously shown that vaccination with the natural tumor peptide Melan-A-induced T cells with superior effector functions as compared with vaccination with the analog peptide optimized for enhanced HLA-A*0201 binding. Here we found that natural peptide vaccination induced tumor-reactive CD8 T cells with frequent coexpression of both memory/homing-associated genes (CD27, IL7R, EOMES, CXCR3, and CCR5) and effector-related genes (IFNG, KLRD1, PRF1, and GZMB), comparable with protective Epstein-Barr virus-specific and cytomegalovirus-specific T cells. In contrast, memory/homing-associated and effector-associated genes were less frequently coexpressed after vaccination with the analog peptide. Remarkably, these findings reveal a previously unknown level of gene expression diversity among vaccine-specific and virus-specific T cells with the simultaneous coexpression of multiple memory/homing-related and effector-related genes by the same cell. Such broad functional gene expression signatures within antigen-specific T cells may be critical for mounting efficient responses to pathogens or tumors. In summary, direct ex vivo high-resolution molecular characterization of individual T cells provides key insights into the processes shaping the functional properties of tumor-specific and virus-specific T cells.

Glycogen metabolism as a marker of astrocyte differentiation.

Glycogen is a hallmark of mature astrocytes, but its emergence during astrocytic differentiation is unclear. Differentiation of E14 mouse neurospheres into astrocytes was induced with fetal bovine serum (FBS), Leukemia Inhibitory Factor (LIF), or Ciliary Neurotrophic Factor (CNTF). Cytochemical and enzymatic analyses showed that glycogen is present in FBS- or LIF- but not in CNTF-differentiated astrocytes. Glycogenolysis was induced in FBS- and LIF-differentiated astrocytes but glycogen resynthesis was observed only with FBS. Protein targeting to glycogen mRNA expression appeared with glial fibrillary acidic protein and S100beta in FBS and LIF conditions but not with CNTF. These results show that glycogen metabolism constitutes a useful marker of astrocyte differentiation.

T cell affinity regulates asymmetric division, effector cell differentiation, and tissue pathology.

The strength of interactions between T cell receptors and the peptide-major histocompatibility complex (pMHC) directly modulates T cell fitness, clonal expansion, and acquisition of effector properties. Here we show that asymmetric T cell division is an important mechanistic link between increased signal strength, effector differentiation, and the ability to induce tissue pathology. Recognition of pMHC above a threshold affinity drove responding T cells into asymmetric cell division. The ensuing proximal daughters underwent extensive division and differentiated into short-lived effector cells expressing the integrin VLA-4, allowing the activated T cell to infiltrate and mediate destruction of peripheral target tissues. In contrast, T cells activated by below-threshold antigens underwent symmetric division, leading to abortive clonal expansion and failure to fully differentiate into tissue-infiltrating effector cells. Antigen affinity and asymmetric division are important factors that regulate fate specification in CD8(+) T cells and predict the potential of a self-reactive T cell to mediate tissue pathology.

The Role of Interleukin-2 in Memory CD8 Cell Differentiation

The current literature on the role of interleukin (IL)-2 in memory CD8(+) T-cell differentiation indicates a significant contribution of IL-2 during primary and also secondary expansion of CD8(+) T cells. IL-2 seems to be responsible for optimal expansion and generation of effector functions following primary antigenic challenge. As the magnitude of T-cell expansion determines the numbers of memory CD8(+) T cells surviving after pathogen elimination, these events influence memory cell generation. Moreover, during the contraction phase of an immune response where most antigen-specific CD8(+) T cells disappear by apoptosis, IL-2 signals are able to rescue CD8(+) T cells from cell death and provide a durable increase in memory CD8(+) T-cell counts. At the memory stage, CD8(+) T-cell frequencies can be boosted by administration of exogenous IL-2. Significantly, only CD8(+) T cells that have received IL-2 signals during initial priming are able to mediate efficient secondary expansion following renewed antigenic challenge. Thus, IL-2 signals during different phases of an immune response are key in optimizing CD8(+) T-cell functions, thereby affecting both primary and secondary responses of these T cells.

Expression hierarchy of T cell epitopes from melanoma differentiation antigens: unexpected high level presentation of tyrosinase-HLA-A2 Complexes revealed by peptide-specific, MHC-restricted, TCR-like antibodies.

Peptide Ags presented by class I MHC molecules on human melanomas and that are recognized by CD8(+) T cells are the subjects of many studies of antitumor immunity and represent attractive candidates for therapeutic approaches. However, no direct quantitative measurements exist to reveal their expression hierarchy on the cell surface. Using novel recombinant Abs which bind these Ags with a peptide-specific, MHC-restricted manner, we demonstrate a defined pattern of expression hierarchy of peptide-HLA-A2 complexes derived from three major differentiation Ags: gp100, Melan-A/Mart-1, and tyrosinase. Studying melanoma cell lines derived from multiple patients, we reveal a surprisingly high level of presentation of tyrosinase-derived complexes and moderate to very low expression of complexes derived from other Ags. No correlation between Ag presentation and mRNA expression was found; however, protein stability may play a major role. These results provide new insights into the characteristics of Ag presentation and are particularly important when such targets are being considered for immunotherapy. These results may shed new light on relationships between Ag presentation and immune response to cancer Ags.

Post-Plutonic Magmatism: from the Source to the Emplacement of an Upper Crustal Dyke Swarm, Adamello Massif, Italy

Magmas of the arc-tholeiitic and calc-alkaline differentiation suites contribute substantially to the formation of continental crust in subduction zones. Different geochemical-petrological models have been put forward to achieve evolved magmas forming large volumes of tonalitic to granitic plutons, building an important part of the continental crust. Primary magmas produced in the mantle wedge overlying the subducted slab migrate through the mantle and the crust. During the transfer, magma can accumulate in intermediate reservoirs at different levels where crystallization leads to differentiation and the heat transfer from the magma, together with gained heat from solidification, lead to partial melting of the crust. Partial melts can be assimilated and mix with more primitive magma. Moreover, already formed crystal cumulates or crystal mushes can be recycled and reactivated to transfer to higher crustal levels. Magma transport in the crust involves fow through fractures within a brittle elastic rock. The solidified magma filled crack, a dyke, can crosscut previously formed geological structures and thus serves as a relative or absolute time marker. The study area is situated in the Adamello massif. The Adamello massif is a composite of plutons that were emplaced between 42 and 29 million years. A later dyke swarm intruded into the southern part of the Adamello Batholith. A fractionation model covering dyke compositions from picrobasalts to dacites results in the cummulative crystallization of 17% olivine, 2% Cr-rich spinel, 18% clinopyroxene, 41% amphibole, 4% plagioclase and 0.1% magnetite to achieve an andesitic composition out of a hydrous primitive picrobasalt. These rocks show a similar geochemical evolution as experimental data simulating fractional crystallization and associated magma differentiation at lower crustal depth (7-10 kbar). The peraluminous, corundum normative composition is one characteristic of more evolved dacitic magmas, which has been explained in a long lasting debate with two di_erent models. Melting of mafic crust or politic material provides one model, whereas an alternative is fractionation from primary mantle derived melts. Amphibole occurring in basaltic-andesitic and andesitic dyke rocks as fractionating cumulate phase extracted from lower crustal depth (6-7.5 kbar) is driving the magmas to peraluminous, corundum normative compositions, which are represented by tonalites forming most of the Adamello Batholith. Most primitive picrobasaltic dykes have a slightly steepened chondrite normalized rare earth elements (REE) pattern and the increased enrichment of light-REE (LREE) for andesites and dacites can be explained by the fractional crystallization model originating from a picrobasalt, taking the changing fractionating phase assemblage and temperature into account. The injection of hot basaltic magma (~1050°C) in a closely spaced dyke swarm increases the surface of the contact to the mainly tonalitic wallrock. Such a setting induces partial melting of the wall rock and selective assimilation. Partial melting of the tonalite host is further expressed through intrusion breccias from basaltic dykes. Heat conduction models with instantaneous magma injection for such a dyke swarm geometry can explain features of partial melting observed in the field. Geochemical data of minerals and bulk rock further underline the selective or bulk assimilation of the tonalite host rock at upper crustal levels (~2-3 kbar), in particular with regard to light ion lithophile elements (LILE) such as Sr, Ba and Rb. Primitive picrobasalts carry an immiscible felsic assimilant as enclaves that bring along refractory rutile and zircon with textures typically found in oceanic plagiogranites or high pressure/low-temperature metamorphic rocks in general. U-Pb data implies a lower Cretaceous age for zircon not yet described as assimilant in Eocene to Oligocene magmatic rocks of the Central Southern Alps. The distribution of post-plutonic dykes in large batholiths such as the Adamello is one of the key features for understanding the regional stress field during the post-batholith emplacement cooling history. The emplacement of the regional dyke swarm covering the southern part of the Adamello massif was associated with consistent left lateral strike-slip movement along magma dilatation planes, leading to en echelon segmentation of dykes. Through the dilation by magma of pre-existing weaknesses and cracks in an otherwise uniform host rock, the dyke propagation and according orientation in the horizontal plane adjusted continuously perpendicular to least compressive remote stress σ3, resulting in an inferred rotation of the remote principal stress field. Les magmas issus des zones de subduction contribuent substantiellement à la formation de la croûte continentale. Les plutons tonalitiques et granitiques représentent, en effet, une partie importante de la croûte continentale. Des magmas primaires produits dans le 'mantle wedge ', partie du manteau se trouvant au-dessus de la plaque plongeante dans des zones de subduction, migrent à travers le manteau puis la croûte. Pendant ce transfert, le magma peut s'accumuler dans des réservoirs intermédiaires à différentes profondeurs. Le stockage de magma dans ces réservoirs engendre, d'une part, la différentiation des magmas par cristallisation fractionnée et, d'autre part, une fusion partielle la croûte continentale préexistante associée au transfert de la chaleur des magmas vers l'encaissant. Ces liquides magmatiques issus de la croûte peuvent, ensuite, se mélanger avec des magmas primaires. Le transport du magma dans la croûte implique notamment un flux de magma à travers différentes fractures recoupant les roches encaissantes élastiques. Au cours de ce processus de migration, des cumulats de cristaux ou des agrégats de cristaux encore non-solidifiés, peuvent être recyclés et réactivés pour être transportés à des niveaux supérieures de la croûte. Le terrain d'étude est situé dans le massif d'Adamello. Celui-ci est composé de plusieurs plutons mis en place entre 42 et 29 millions d'années. Dans une phase tardive de l'activité magmatique liée à ce batholite, une série de filons de composition variable allant de picrobasalte à des compositions dacitiques s'est mise en place la partie sud du massif. Deux modèles sont proposés dans la littérature, pour expliquer la formation des magmas dacitiques caractérisés par des compositions peralumineux (i.e. à corindon normatif). Le premier modèle propose que ces magmas soient issus de la fusion de matériel mafique et pélitique présent dans la partie inférieur de la croûte, alors que le deuxième modèle suggère une évolution par cristallisation fractionnée à partir de liquides primaires issus du manteau. Un modèle de cristallisation fractionnée a pu être développé pour expliquer l'évolution des filons de l'Adamello. Ce modèle explique la formation des filons dacitiques par la cristallisation fractionnée de 17% olivine, 2% spinelle riche en Cr, 18% clinopyroxène, 41% amphibole, 4% plagioclase et 0.1% magnetite à partir de liquide de compositions picrobasaltiques. Ce modèle prend en considération les contraintes pétrologiques déduites de l'observation des différents filons ainsi que du champ de stabilité des différentes phases en fonction de la température. Ces roches montrent une évolution géochimique similaire aux données expérimentales simulant la cristallisation fractionnée de magmas évoluant à des niveaux inférieurs de la croûte (7-10 kbar). Le modèle montre, en particulier, le rôle prépondérant de l'amphibole, une phase qui contrôle en particulier le caractère peralumineux des magmas différentiés ainsi que leurs compositions en éléments en traces. Des phénomènes de fusion partielle de l'encaissant tonalitique lors de la mise en place de _lons mafiques sont observée sur le terrain. L'injection du magma basaltique chaud (~1050°C) sous forme de filons rapprochés augmente la surface du contact avec l'encaissante tonalitique. Une telle situation produit la fusion partielle des roches encaissantes nécessaire à l'incorporation d'enclaves mafiques observés au sein des tonalites. Pour comprendre les conditions nécessaires pour la fusion partielle des roches encaissantes, des modèles de conduction thermique pour une injection simultanée d'une série de filons ont été développées. Des données géochimiques sur les minéraux et sur les roches totales soulignent qu'au niveau supérieur de la croûte, l'assimilation sélective ou totale de l'encaissante tonalitique modifie la composition du liquide primaire pour les éléments lithophiles tel que le Sr, Ba et Rb. Un autre aspect important concernant la pétrologie des filons de l'Adamello est la présence d'enclaves felsiques dans les filons les plus primitifs. Ces enclaves montrent, en particulier, des textures proches de celles rencontrées dans des plagiogranites océaniques ou dans des roches métamorphiques de haute pression/basse température. Ces enclaves contiennent du zircon et du rutile. La datations de ces zircons à l'aide du géochronomètre U-Pb indique un âge Crétacé inférieur. Cet âge est important, car aucune roche de cet âge n'a été considérée comme un assimilant potentiel pour des roches magmatiques d'âge Eocène à Oligocène dans les Alpes Sud Centrales. La réparation spatiale des filons post-plutoniques dans des grands batholites tel que l'Adamello, est une caractéristique clé pour la compréhension des champs de contraintes lors du refroidissement du batholite. L'orientation des filons va, en particulier, indiqué la contrainte minimal au sein des roches encaissante. La mise en place de la série de filon recoupant la partie Sud du massif de l'Adamello est associée à un décrochement senestre, un décrochement que l'on peut lié aux contraintes tectoniques régionales auxquelles s'ajoutent l'effet de la dilatation produite par la mise en place du batholite lui-même. Ce décrochement senestre produit une segmentation en échelon des filons.

Cell death in Leishmania induced by stress and differentiation: programmed cell death or necrosis?

Unicellular organisms, such as the protozoan parasite Leishmania, can be stimulated to show some morphological and biochemical features characteristic of mammalian apoptosis. This study demonstrates that under a variety of stress conditions such as serum deprivation, heat shock and nitric oxide, cell death can be induced leading to genomic DNA fragmentation into oligonucleosomes. DNA fragmentation was observed, without induction, in the infectious stages of the parasite, and correlated with the presence of internucleosomal nuclease activity, visualisation of 45 to 59 kDa nucleases and detection of TUNEL-positive nuclei. DNA fragmentation was not dependent on active effector downstream caspases nor on the lysosomal cathepsin L-like enzymes CPA and CPB. These data are consistent with the presence of a caspase-independent cell death mechanism in Leishmania, induced by stress and differentiation that differs significantly from metazoa.

Selective effects of PPARgamma agonists and antagonists on human pre-adipocyte differentiation.

Aim: The insulin sensitizer rosiglitazone (RTZ) acts by activating peroxisome proliferator and activated receptor gamma (PPAR gamma), an effect accompanied in vivo in humans by an increase in fat storage. We hypothesized that this effect concerns PPARgamma(1) and PPARgamma(2) differently and is dependant on the origin of the adipose cells (subcutaneous or visceral). To this aim, the effect of RTZ, the PPARgamma antagonist GW9662 and lentiviral vectors expressing interfering RNA were evaluated on human pre-adipocyte models. Methods: Two models were investigated: the human pre-adipose cell line Chub-S7 and primary pre-adipocytes derived from subcutaneous and visceral biopsies of adipose tissue (AT) obtained from obese patients. Cells were used to perform oil-red O staining, gene expression measurements and lentiviral infections. Results: In both models, RTZ was found to stimulate the differentiation of pre-adipocytes into mature cells. This was accompanied by significant increases in both the PPARgamma(1) and PPARgamma(2) gene expression, with a relatively stronger stimulation of PPARgamma(2). In contrast, RTZ failed to stimulate differentiation processes when cells were incubated in the presence of GW9662. This effect was similar to the effect observed using interfering RNA against PPARgamma(2). It was accompanied by an abrogation of the RTZ-induced PPARgamma(2) gene expression, whereas the level of PPARgamma(1) was not affected. Conclusions: Both the GW9662 treatment and interfering RNA against PPARgamma(2) are able to abrogate RTZ-induced differentiation without a significant change of PPARgamma(1) gene expression. These results are consistent with previous results obtained in animal models and suggest that in humans PPARgamma(2) may also be the key isoform involved in fat storage.