Approche hydrogéochimique et structurale des circulations dans un réservoir du domaine alpin (massif d'Allevard, France)

Les caractéristiques hydrogéochimiques des réservoirs profonds du domaine alpin occidental sont abordées par le biais du système hydrominéral d'Allevard (France). Le volume des données et la variabilité des caractéristiques qui en découle, permettent de considérer cette source comme un exemple représentatif des sources thermominérales du domaine alpin occidental. L'étude combinée de ces données relatives à l'hydrochimie, à la géochimie isotopique, à la présence de gaz et d'éléments traces, confrontées au contexte fissural et structural des réservoirs, permet une approche cohérente des circulations profondes.On propose en conclusion un modèle qui pourrait être généralisé aux systèmes thermominéraux situés dans un contexte structural comparable (front de chevauchement des massifs cristallins externes; front de chevauchement briançonnais) et présentant un faciès semblable (eaux chloro-sulfatées). Ce modèle tient compte de mélanges entre eaux d'origine superficielle et/ou semi-profondes et d'eaux d'origine profonde.Investigations of the hydrogeochemical and structural characteristics of groundwater flows in an Alpine hydrothermal reservoir (Allevard Massif, French Western Alps) are carried out in order to define the origin and genesis of thermal and mineral waters.The geological formations constituting the Allevard Massif consist of a Hercynian basement and a continuous sedimentary cover from Carboniferous to Middle Jurassic. The basement is composed mainly of micaschists, the sedimentary cover of Paleozoic clastic deposits : Triassic formations (evaporites, dolomites, limestones); Liassic calcareous marls, schistics marls and Limestones. A groundwater outlet (Allevard Spring) from this reservoir is located in the calcareous terrains of Lias.The basement-cover contact is a major N40 subvertical fault. Faulting tectonics and subsequent rock fracturing is important for the hydrogeology of the area under study since the geological formations are initially impermeable (except for the Triassic terrains). Observations of underground galleries in the neighbourhood of the spring showed the presence of water outflows located on N40 strike-slip faults affecting both the basement and the Liassic terrains. The role of faulting tectonics in groundwater occurrence is indeed clearly emphasized by the position of the Allevard Spring at the intersection of two major faults with N60 and N140 trends.Available data cover nearly a hundred years (1888-1930) and emphasize the great variability of the chemical features of the spring water. This is related to the tact that the water outflowing from the spring is a mixture in varying proportions of waters from different sources. Combined interpretation of the structural characteristics, isotopical (3H; 18O; 2H; 34S) and chemico-physical (major and trace elements, gases, temperature, ...) data provides the basis of a working assumption on a flow and mixing model in which three different hydrogeological subsystems were distinguished : a surface subsystem comprising cold infiltration waters of a bicarbonate-calcium and a sulfate-calcium type; a subsurface subsystem constituted by the fissured Liassic calcareous terrains comprising part of the infiltrated waters which intermingle with much warmer waters of a sodium-chloride type originating from a subsystem located at a great depth. By means of geochemical indicators, a depth of nearly 4000 m has been estimated.Such a model, in agreement with geological evidence and reported by some recent works showing that halite deposits are probable at great depths, could be generalized to other hydrothermal systems occurring within the framework of the French Western Alps in a comparable structural context (thrust front of the external metamorphic massifs; "Briançonnais" thrust front)

Metal immobilization and soil amendment efficiency at a contaminated sediment landfill site: a field study focusing on plants, springtails, and bacteria

International audienceMetal immobilization may contribute to the environmental management strategy of dredged sediment landfill sites contaminated by metals. In a field experiment, amendment effects and efficiency were investigated, focusing on plants, springtails and bacteria colonisation, metal extractability and sediment ecotoxicity. Conversely to hydroxylapatite (HA, 3% DW), the addition of Thomas Basic Slag (TBS, 5% DW) to a 5-yr deposited sediment contaminated with Zn, Cd, Cu, Pb and As resulted in a decrease in the 0.01 M Ca(NO(3))(2)-extractable concentrations of Cd and Zn. Shoot Cd and Zn concentration in Calamagrostis epigejos, the dominant plant species, also decreased in the presence of TBS. The addition of TBS and HA reduced sediment ecotoxicity and improved the growth of the total bacterial population. Hydroxylapatite improved plant species richness and diversity and decreased antioxidant enzymes in C. Epigejos and Urtica dïoica. Collembolan communities did not differ in abundance and diversity between the different treatments

Trapping DNA Replication Origins from the Human Genome

Synthesis of chromosomal DNA is initiated from multiple origins of replication in higher eukaryotes; however, little is known about these origins’ structures. We isolated the origin-derived nascent DNAs from a human repair-deficient cell line by blocking the replication forks near the origins using two different origin-trapping methods (i.e., UV- or chemical crosslinker-treatment and cell synchronization in early S phase using DNA replication inhibitors). Single-stranded DNAs (of 0.5–3 kb) that accumulated after such treatments were labeled with bromodeoxyuridine (BrdU). BrdU-labeled DNA was immunopurified after fractionation by alkaline sucrose density gradient centrifugation and cloned by complementary-strand synthesis and PCR amplification. Competitive PCR revealed an increased abundance of DNA derived from known replication origins (c-myc and lamin B2 genes) in the nascent DNA fractions from the UV-treated or crosslinked cells. Nucleotide sequences of 85 and 208 kb were obtained from the two libraries (I and II) prepared from the UV-treated log-phase cells and early S phase arrested cells, respectively. The libraries differed from each other in their G+C composition and replication-related motif contents, suggesting that differences existed between the origin fragments isolated by the two different origin-trapping methods. The replication activities for seven out of 12 putative origin loci from the early-S phase cells were shown by competitive PCR. We mapped 117 (library I) and 172 (library II) putative origin loci to the human genome; approximately 60% and 50% of these loci were assigned to the G-band and intragenic regions, respectively. Analyses of the flanking sequences of the mapped loci suggested that the putative origin loci tended to associate with genes (including conserved sites) and DNase I hypersensitive sites; however, poor correlations were found between such loci and the CpG islands, transcription start sites, and K27-acetylated histone H3 peaks

Quantitative assessments of glycolysis from single cells

The most common positron emission tomography (PET) radio-labeled probe for molecular diagnostics in patient care and research is the glucose analog, 2-deoxy-2-[F-18]fluoro-D-glucose (^(18)F-FDG). We report on an integrated microfluidics-chip/beta particle imaging system for in vitro ^(18)F-FDG radioassays of glycolysis with single cell resolution. We investigated the kinetic responses of single glioblastoma cancer cells to targeted inhibitors of receptor tyrosine kinase signaling. Further, we find a weak positive correlation between cell size and rate of glycolysis

Single-Cell Phosphoproteomics Resolves Adaptive Signaling Dynamics and Informs Targeted Combination Therapy in Glioblastoma

Intratumoral heterogeneity of signaling networks may contribute to targeted cancer therapy resistance, including in the highly lethal brain cancer glioblastoma (GBM). We performed single-cell phosphoproteomics on a patient-derived in vivo GBM model of mTOR kinase inhibitor resistance and coupled it to an analytical approach for detecting changes in signaling coordination. Alterations in the protein signaling coordination were resolved as early as 2.5 days after treatment, anticipating drug resistance long before it was clinically manifest. Combination therapies were identified that resulted in complete and sustained tumor suppression in vivo. This approach may identify actionable alterations in signal coordination that underlie adaptive resistance, which can be suppressed through combination drug therapy, including non-obvious drug combinations

Common fragile sites are characterized by histone hypoacetylation

Common fragile sites (CFSs) represent large, highly unstable regions of the human genome. CFS sequences are sensitive to perturbation of replication; however, the molecular basis for the instability at CFSs is poorly understood. We hypothesized that a unique epigenetic pattern may underlie the unusual sensitivity of CFSs to replication interference. To examine this hypothesis, we analyzed chromatin modification patterns within the six human CFSs with the highest levels of breakage, and their surrounding non-fragile regions (NCFSs). Chromatin at most of the CFSs analyzed has significantly less histone acetylation than that of their surrounding NCFSs. Trichostatin A and/or 5-azadeoxycytidine treatment reduced chromosome breakage at CFSs. Furthermore, chromatin at the most commonly expressed CFS, the FRA3B, is more resistant to micrococcal nuclease than that of the flanking non-fragile sequences. These results demonstrate that histone hypoacetylation is a characteristic epigenetic pattern of CFSs, and chromatin within CFSs might be relatively more compact than that of the NCFSs, indicating a role for chromatin conformation in genomic instability at CFSs. Moreover, lack of histone acetylation at CFSs may contribute to the defective response to replication stress characteristic of CFSs, leading to the genetic instability characteristic of this regions

Quantitative assessments of glycolysis from single cells

The most common positron emission tomography (PET) radio-labeled probe for molecular diagnostics in patient care and research is the glucose analog, 2-deoxy-2-[F-18]fluoro-D-glucose (^(18)F-FDG). We report on an integrated microfluidics-chip/beta particle imaging system for in vitro ^(18)F-FDG radioassays of glycolysis with single cell resolution. We investigated the kinetic responses of single glioblastoma cancer cells to targeted inhibitors of receptor tyrosine kinase signaling. Further, we find a weak positive correlation between cell size and rate of glycolysis

Physiological and anthocyanin biosynthesis genes response induced by vanadium stress in mustard genotypes with distinct photosynthetic activity

The present study aimed to elucidate the photosynthetic performance, antioxidant enzyme activities, anthocyanin contents, anthocyanin biosynthetic gene expression, and vanadium uptake in mustard genotypes (purple and green) that differ in photosynthetic capacity under vanadium stress. The results indicated that vanadium significantly reduced photosynthetic activity in both genotypes. The activities of the antioxidant enzymes were increased significantly in response to vanadium in both genotypes, although the purple exhibited higher. The anthocyanin contents were also reduced under vanadium stress. The anthocyanin biosynthetic genes were highly expressed in the purple genotype, notably the genes TT8, F3H, and MYBL2 under vanadium stress. The results indicate that induction of TT8, F3H, and MYBL2 genes was associated with upregulation of the biosynthetic genes required for higher anthocyanin biosynthesis in purple compared with the green mustard. The roots accumulated higher vanadium than shoots in both mustard genotypes. The results indicate that the purple mustard had higher vanadium tolerance

Evidence for Sequential and Increasing Activation of Replication Origins along Replication Timing Gradients in the Human Genome

Genome-wide replication timing studies have suggested that mammalian chromosomes consist of megabase-scale domains of coordinated origin firing separated by large originless transition regions. Here, we report a quantitative genome-wide analysis of DNA replication kinetics in several human cell types that contradicts this view. DNA combing in HeLa cells sorted into four temporal compartments of S phase shows that replication origins are spaced at 40 kb intervals and fire as small clusters whose synchrony increases during S phase and that replication fork velocity (mean 0.7 kb/min, maximum 2.0 kb/min) remains constant and narrowly distributed through S phase. However, multi-scale analysis of a genome-wide replication timing profile shows a broad distribution of replication timing gradients with practically no regions larger than 100 kb replicating at less than 2 kb/min. Therefore, HeLa cells lack large regions of unidirectional fork progression. Temporal transition regions are replicated by sequential activation of origins at a rate that increases during S phase and replication timing gradients are set by the delay and the spacing between successive origin firings rather than by the velocity of single forks. Activation of internal origins in a specific temporal transition region is directly demonstrated by DNA combing of the IGH locus in HeLa cells. Analysis of published origin maps in HeLa cells and published replication timing and DNA combing data in several other cell types corroborate these findings, with the interesting exception of embryonic stem cells where regions of unidirectional fork progression seem more abundant. These results can be explained if origins fire independently of each other but under the control of long-range chromatin structure, or if replication forks progressing from early origins stimulate initiation in nearby unreplicated DNA. These findings shed a new light on the replication timing program of mammalian genomes and provide a general model for their replication kinetics