480 research outputs found

    Cinétiques de biodégradation par boues activées de la matière organique soluble d'un effluent synthétique

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    L'approche expérimentale choisie pour cette étude a eu pour objet de mesurer en conditions batch, les cinétiques d'élimination de la demande chimique en oxygène soluble d'un effluent synthétique mis en contact avec des boues activées d'origine différente. Les essais conduits en laboratoire ont été réalisés en faisant varier le rapport So/Xo (mg de DCO initiale par mg de matières volatiles initiales) telles que les concentrations en So et en Xo correspondent aux concentrations en DCO soluble (DCOs) et en matières volatiles (MV) rencontrées sur les stations d'épuration. Les essais ont été effectués sous aération continue à 20 °C en mettant en contact l'effluent synthétique et de la boue activée prélevée depuis 24 h et stockée à 4 °C dans l'attente de l'essai. De ce fait, la valeur de So mesurée au début de l'essai représente la concentration en DCO amenée par l'effluent synthétique (Seff) et celle amenée par l'inoculum de boue activée (Sb) représentant selon les essais de 5 à 70 % de la DCO de l'essai. Les profils de cinétique d'élimination de la DCO soluble obtenus pour différentes conditions d'essai s'ajustent, selon les valeurs de So/Xo (So/Xo variant de 0,15 à 2,17 ) à une fonction du premier ordre par rapport au substrat ou à une fonction sigmoïde. Le type de fonction cinétique d'élimination est également contrôlé par la proportion de la DCO amenée par l'inoculum.The conventional activated sludge process used for wastewater treatment removes from 80 to 95% of the total organic matter. However, a quantity of "not well identified" (particular, colloidal and soluble) organic matter is always present in the treated effluent. Reducing this residual (and improving the treatment efficiency) requires knowledge of the origin of that organic matter and especially to determine the fraction originating from the influent and the fraction generated by the biomass.This research has been conducted in batch conditions and studies the soluble COD (CODs) removal kinetics of a synthetic effluent (casein + starch + acetate + mineral salts), in contact with different activated sludge originating from six different wastewater treatment plants (loads varying from 0.06 to 1.14 kg BOD[inf]5/kg VSS. d).Experiments have been conducted with different So/Xo values (ratio between CODs initial concentration and VSS initial concentration) in order that these values correspond to the CODs and VSS values found in the plants.In accordance with GRAU et al. (1975), CECH and CHUDOBA (1983), PITTER and CHUDOBA (1990), CHUDOBA et al (1992), the So/Xo ratio is a fundamental parameter governing the kinetics reactions.Experiments have been conducted under continuous aeration at 20°C where the synthetic wastewater (500 ml) is in contact with activated sludge (200 ml) collected 24 h before and stored at 4°C until the batch is started. In this manner, the initial So is due to the CODs of the synthetic effluent (So eff=197 mg/l) and to the CODs originating from the sludge (5 to 70% of the initial CODs). The initial VSS concentration (Xo) is between 0.6 and 2.5 g/l. Kinetics of CODs removal are simulated by two functions: the first order function, where the initial rate is the maximal, and the sigmoidal function where the maximal rate is reached after a lag time (3 to 8 h).Concerning the first order functions, the degradation rate is faster when the ratio is low (So/Xo lower than 0.44). This is not the case for the sigmoidal functions. In the results of this study, the residual of COD is always lower when the degradation kinetic follows the exponential model.Our experiments show that the type of degradation kinetics (first order or sigmoidal) is not only controlled by the So/Xo parameter but also by the proportion of CODs brought by the sludge and that parameter can play a determinant role.When the proportion of CODs brought by the sludge is very large (between 41% to 46%) the degradation reactions follow the sigmoidal type. These results can possibly be explained by the low biodegradability of the polymers or molecules originating from the inoculum which has been stored during 24 h, or by the low activity of the biomass after 24 h of storage on the biodegradation of the soluble organic matte

    Genome Expression Profile Analysis of the Immature Maize Embryo during Dedifferentiation

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    Maize is one of the most important cereal crops worldwide and one of the primary targets of genetic manipulation, which provides an excellent way to promote its production. However, the obvious difference of the dedifferentiation frequency of immature maize embryo among various genotypes indicates that its genetic transformation is dependence on genotype and immature embryo-derived undifferentiated cells. To identify important genes and metabolic pathways involved in forming of embryo-derived embryonic calli, in this study, DGE (differential gene expression) analysis was performed on stages I, II, and III of maize inbred line 18-599R and corresponding control during the process of immature embryo dedifferentiation. A total of ∼21 million cDNA tags were sequenced, and 4,849,453, 5,076,030, 4,931,339, and 5,130,573 clean tags were obtained in the libraries of the samples and the control, respectively. In comparison with the control, 251, 324 and 313 differentially expressed genes (DEGs) were identified in the three stages with more than five folds, respectively. Interestingly, it is revealed that all the DEGs are related to metabolism, cellular process, and signaling and information storage and processing functions. Particularly, the genes involved in amino acid and carbohydrate transport and metabolism, cell wall/membrane/envelope biogenesis and signal transduction mechanism have been significantly changed during the dedifferentiation. To our best knowledge, this study is the first genome-wide effort to investigate the transcriptional changes in dedifferentiation immature maize embryos and the identified DEGs can serve as a basis for further functional characterization

    Development and quantitative analyses of a universal rRNA-subtraction protocol for microbial metatranscriptomics

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    Metatranscriptomes generated by pyrosequencing hold significant potential for describing functional processes in complex microbial communities. Meeting this potential requires protocols that maximize mRNA recovery by reducing the relative abundance of ribosomal RNA, as well as systematic comparisons to identify methodological artifacts and test for reproducibility across data sets. Here, we implement a protocol for subtractive hybridization of bacterial rRNA (16S and 23S) that uses sample-specific probes and is applicable across diverse environmental samples. To test this method, rRNA-subtracted and unsubtracted transcriptomes were sequenced (454 FLX technology) from bacterioplankton communities at two depths in the oligotrophic open ocean, yielding 10 data sets representing ~350 Mbp. Subtractive hybridization reduced bacterial rRNA transcript abundance by 40–58%, increasing recovery of non-rRNA sequences up to fourfold (from 12% to 20% of total sequences to 40–49%). In testing this method, we established criteria for detecting sequences replicated artificially via pyrosequencing errors and identified such replicates as a significant component (6–39%) of total pyrosequencing reads. Following replicate removal, statistical comparisons of reference genes (identified via BLASTX to NCBI-nr) between technical replicates and between rRNA-subtracted and unsubtracted samples showed low levels of differential transcript abundance (<0.2% of reference genes). However, gene overlap between data sets was remarkably low, with no two data sets (including duplicate runs from the same pyrosequencing library template) sharing greater than 17% of unique reference genes. These results indicate that pyrosequencing captures a small subset of total mRNA diversity and underscores the importance of reliable rRNA subtraction procedures to enhance sequencing coverage across the functional transcript pool.Agouron InstituteGordon and Betty Moore FoundationUnited States. Dept. of Energy. Office of ScienceNational Science Foundation (U.S.) (NSF Science and Technology Center Award EF0424599

    Tissue Type-Specific Expression of the dsRNA-Binding Protein 76 and Genome-Wide Elucidation of Its Target mRNAs

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    Background: RNA-binding proteins accompany all steps in the life of mRNAs and provide dynamic gene regulatory functions for rapid adjustment to changing extra-or intracellular conditions. The association of RNA-binding proteins with their targets is regulated through changing subcellular distribution, post-translational modification or association with other proteins. Methodology: We demonstrate that the dsRNA binding protein 76 (DRBP76), synonymous with nuclear factor 90, displays inherently distinct tissue type-specific subcellular distribution in the normal human central nervous system and in malignant brain tumors of glial origin. Altered subcellular localization and isoform distribution in malignant glioma indicate that tumor-specific changes in DRBP76-related gene products and their regulatory functions may contribute to the formation and/or maintenance of these tumors. To identify endogenous mRNA targets of DRBP76, we performed RNA-immunoprecipitation and genome-wide microarray analyses in HEK293 cells, and identified specific classes of transcripts encoding critical functions in cellular metabolism. Significance: Our data suggest that physiologic DRBP76 expression, isoform distribution and subcellular localization are profoundly altered upon malignant transformation. Thus, the functional role of DRBP76 in co- or post-transcriptional gene regulation may contribute to the neoplastic phenotype

    The Transcriptional Response to DNA-Double-Strand Breaks in Physcomitrella patens

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    The model bryophyte Physcomitrella patens is unique among plants in supporting the generation of mutant alleles by facile homologous recombination-mediated gene targeting (GT). Reasoning that targeted transgene integration occurs through the capture of transforming DNA by the homology-dependent pathway for DNA double-strand break (DNA-DSB) repair, we analysed the genome-wide transcriptomic response to bleomycin-induced DNA damage and generated mutants in candidate DNA repair genes. Massively parallel (Illumina) cDNA sequencing identified potential participants in gene targeting. Transcripts encoding DNA repair proteins active in multiple repair pathways were significantly up-regulated. These included Rad51, CtIP, DNA ligase 1, Replication protein A and ATR in homology-dependent repair, Xrcc4, DNA ligase 4, Ku70 and Ku80 in non-homologous end-joining and Rad1, Tebichi/polymerase theta, PARP in microhomology-mediated end-joining. Differentially regulated cell-cycle components included up-regulated Rad9 and Hus1 DNA-damage-related checkpoint proteins and down-regulated D-type cyclins and B-type CDKs, commensurate with the imposition of a checkpoint at G2 of the cell cycle characteristic of homology-dependent DNA-DSB repair. Candidate genes, including ATP-dependent chromatin remodelling helicases associated with repair and recombination, were knocked out and analysed for growth defects, hypersensitivity to DNA damage and reduced GT efficiency. Targeted knockout of PpCtIP, a cell-cycle activated mediator of homology-dependent DSB resection, resulted in bleomycin-hypersensitivity and greatly reduced GT efficiency

    Screening and identification of seed-specific genes using digital differential display tools combined with microarray data from common wheat

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    <p>Abstract</p> <p>Background</p> <p>Wheat is one of the most important cereal crops for human beings, with seeds being the tissue of highly economic value. Various morphogenetic and metabolic processes are exclusively associated with seed maturation. The goal of this study was to screen and identify genes specifically expressed in the developing seed of wheat with an integrative utilization of digital differential display (DDD) and available online microarray databases.</p> <p>Results</p> <p>A total of 201 unigenes were identified as the results of DDD screening and microarray database searching. The expressions of 6 of these were shown to be seed-specific by qRT-PCR analysis. Further GO enrichment analysis indicated that seed-specific genes were mainly associated with defense response, response to stress, multi-organism process, pathogenesis, extracellular region, nutrient reservoir activity, enzyme inhibitor activity, antioxidant activity and oxidoreductase activity. A comparison of this set of genes with the rice (<it>Oryza sativa</it>) genome was also performed and approximately three-fifths of them have rice counterparts. Between the counterparts, around 63% showed similar expression patterns according to the microarray data.</p> <p>Conclusions</p> <p>In conclusion, the DDD screening combined with microarray data analysis is an effective strategy for the identification of seed-specific expressed genes in wheat. These seed-specific genes screened during this study will provide valuable information for further studies about the functions of these genes in wheat.</p

    Wig-1, a novel regulator of N-Myc mRNA and N-Myc-driven tumor growth

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    Wig-1 is a transcriptional target of the p53 tumor suppressor and encodes an mRNA stability-regulating protein. We show here that Wig-1 knockdown causes a dramatic inhibition of N-Myc expression and triggers differentiation in neuroblastoma cells carrying amplified N-Myc. Transient Wig-1 knockdown significantly delays development of N-Myc-driven tumors in mice. We also show that N-Myc expression is induced upon moderate p53-activating stress, suggesting a role of the p53-Wig-1-N-Myc axis in promoting cell cycle re-entry upon p53-induced cell cycle arrest and DNA repair. Moreover, our findings raise possibilities for the improved treatment of poor prognosis neuroblastomas that carry amplified N-Myc

    Transcriptome analysis of mRNA and miRNA in skeletal muscle indicates an important network for differential Residual Feed Intake in pigs

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    Feed efficiency (FE) can be measured by feed conversion ratio (FCR) or residual feed intake (RFI). In this study, we measured the FE related phenotypes of 236 castrated purebred Yorkshire boars, and selected 10 extreme individuals with high and low RFI for transcriptome analysis. We used RNA-seq analyses to determine the differential expression of genes and miRNAs in skeletal muscle. There were 99 differentially expressed genes identified (q ≤ 0.05). The down-regulated genes were mainly involved in mitochondrial energy metabolism, including FABP3, RCAN, PPARGC1 (PGC-1A), HK2 and PRKAG2. The up-regulated genes were mainly involved in skeletal muscle differentiation and proliferation, including IGF2, PDE7A, CEBPD, PIK3R1 and MYH6. Moreover, 15 differentially expressed miRNAs (|log2FC| ≥ 1, total reads count ≥ 20, p ≤ 0.05) were identified. Among them, miR-136, miR-30e-5p, miR-1, miR-208b, miR-199a, miR-101 and miR-29c were up-regulated, while miR-215, miR-365-5p, miR-486, miR-1271, miR-145, miR-99b, miR-191 and miR-10b were down-regulated in low RFI pigs. We conclude that decreasing mitochondrial energy metabolism, possibly through AMPK - PGC-1A pathways, and increasing muscle growth, through IGF-1/2 and TGF-β signaling pathways, are potential strategies for the improvement of FE in pigs (and possibly other livestock). This study provides new insights into the molecular mechanisms that determine RFI and FE in pigs

    Altered Levels of Histone Deacetylase OsHDT1 Affect Differential Gene Expression Patterns in Hybrid Rice

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    Hybrids between different inbred varieties display novel patterns of gene expression resulted from parental variation in allelic nucleotide sequences. To study the function of chromatin regulators in hybrid gene expression, the histone deacetylase gene OsHDT1 whose expression displayed a circadian rhythm was over-expressed or inactivated by RNAi in an elite rice parent. Increased OsHDT1 expression did not affect plant growth in the parent but led to early flowering in the hybrid. Nonadditive up-regulation of key flowering time genes was found to be related to flowering time of the hybrid. Over-expression of OsHDT1 repressed the nonadditive expression of the key flowering repressors in the hybrid (i.e. OsGI and Hd1) inducing early flowering. Analysis of histone acetylation suggested that OsHDT1 over-expression might promote deacetylation on OsGI and Hd1 chromatin during the peak expression phase. High throughput differential gene expression analysis revealed that altered OsHDT1 levels affected nonadditive expression of many genes in the hybrid. These data demonstrate that nonadditive gene expression was involved in flowering time control in the hybrid rice and that OsHDT1 level was important for nonadditive or differential expression of many genes including the flowering time genes, suggesting that OsHDT1 may be involved in epigenetic control of parental genome interaction for differential gene expression
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