53161 research outputs found
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Detection of unknown genetically modified organisms (GMO) by statistical analysis of high throughput sequencing data
No full textSystematic analysis of truseq, smarter and smarter ultra-Lowrna-seq kits for standard, low and ultra-low quantity samples
No full textHigh-throughput RNA-sequencing has become the gold standard method for whole-transcriptome gene expression analysis, and is widely used in numerous applications to study cell and tissue transcriptomes. It is also being increasingly used in a number of clinical applications, including expression profiling for diagnostics and alternative transcript detection. However, despite its many advantages, RNA sequencing can be challenging in some situations, for instance in cases of low input amounts or degraded RNA samples. Several protocols have been proposed to overcome these challenges, and many are available as commercial kits. In this study, we systematically test three recent commercial technologies for RNA-seq library preparation (TruSeq, SMARTer and SMARTer Ultra-Low) on human biological reference materials, using standard (1 mg), low (100 ng and 10 ng) and ultra-low (<1 ng) input amounts, and for mRNA and total RNA, stranded and unstranded. The results are analyzed using read quality and alignment metrics, gene detection and differential gene expression metrics. Overall, we show that the TruSeq kit performs well with an input amount of 100 ng, while the SMARTer kit shows decreased performance for inputs of 100 and 10 ng, and the SMARTer Ultra-Low kit performs relatively well for input amounts <1 ng. All the results are discussed in detail, and we provide guidelines for biologists for the selection of an RNA-seq library preparation kit
Identification of extracellular vesicles involved in the biomineralization of the hen eggshell
No full textThe preference‐performance relationship as a means of classifying parasitoids according to their specialization degree
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Cryptosporidium parvum can subvert the host immune response through manipulation of CRAMP expression during neonatal infection
No full textDue to the immaturity of their immune system, neonates are highly sensitive to intestinal infections. During the neonatal period, antimicrobial peptide (AMP) composition differs substantially from that of adults. This is the case in the small intestine for the cathelicidin-related antimicrobial peptide (CRAMP) expressed preferentially in the neonatal period while conversely other AMPs such as Reg3γ are expressed later in life. Among enteric neonatal diseases, Cryptosporidiosis is a zoonotic disease and is highly prevalent in children less than 5 years old in developing countries and in neonatal ruminants worldwide. Cryptosporidium parvum is the etiological agent of this diarrheal disease and infects exclusively epithelial cells. Innate immunity is important to control the acute phase of infection in neonates with dendritic cells and IFNγ playing a major role. Antimicrobial peptides are important contributors of innate immunity, but the role of CRAMP, which is elevated in the intestine of neonates has never been investigated during Cryptosporidiosis so far. In this work, we observed in the neonatal murine model of cryptosporidiosis that unlike other antimicrobial molecules such as Reg3 and Lysozyme, CRAMP expression was significantly reduced in the intestine during infection. By using different genetically modified mouse models, we demonstrated that the reduced CRAMP expression was independent of IFN , a pro-inflammatory cytokine strongly produced during infection, but also of Myd88, an adaptor molecule involved in innate immune signalling. We also excluded the role of gut flora in this response. When C. parvum infected neonatal mice orally received exogenous CRAMP to compensate the reduced expression of this AMP, the parasitic load of neonates was significantly decreased. In addition, when free parasites were in direct contact with CRAMP, this AMP affected the viability of sporozoites. All together, these data suggest that C. parvum induces the reduction of CRAMP expression to escape the anti-parasiticidal effect of CRAMP
Intelligence professionnelle du formateur-accompagnateur de transitions professionnelles : activités d’adaptation et de coopération dans quatre dispositifs d’accompagnement
No full textA partir de notre expérience de quatre dispositifs visant la reconfiguration de l’activité dans le
cadre de transitions professionnelles, nous chercherons à éclairer l’activité du
formateur/accompagnateur dans ses dimensions adaptative, coopérative et attentive à la part
sensible et émotionnelle des situations. Nous porterons un regard dans ces expériences sur
l’articulation entre diagnostic, animation et évaluation, en nous inspirant du concept de
diagnostic en situation dynamique (Hoc et Amalberti, 1994) et sur l’intelligence professionnelle
collective pour mener, adapter et articuler ces différentes étapes à plusieurs
Projet RELAPA (génomique pour la REsistance génétique des LApins a la PAsteurellose) : paramètres génétiques
No full textCette étude est une première approche d’estimation de paramètres génétiques de résistance à la Pasteurellose par infection expérimentale à Pasteurella multocida d’une population de lapins croisés. De nouveaux caractères de résistance ont été étudiés : une note de dissémination des abcès enregistrée à l’autopsie, une note de présence ou absence de Pasteurella dans les organes et une note globale de résistance résumant les deux précédentes. Pour ces caractères, l’héritabilité varie de 0,09 (±0,05) à 0,16 (±0,06). L’héritabilité la plus importante a été estimée pour la note de résistance. Ces estimations suggèrent la contribution d’une composante génétique à la résistance à la pasteurellose. Les corrélations génétiques entre la note de résistance et les différentes mesures de croissance sont élevées et positives, de 0,70 (±0,14) à 0,98 (±0,06). Il serait donc possible de mettre en place une sélection sur la résistance à la Pasteurellose dans les schémas de sélection de lapins de chair français basée sur un de ces critères. Des études complémentaires doivent encore être conduites sur d’autres caractères de production et de de résistance à la Pasteurellos
Anti-VEGF signalling mechanism in HUVECs by melatonin, serotonin, hydroxytyrosol and other bioactive compounds
No full textAngiogenesis drives evolution and destabilisation of atherosclerotic plaques and the growth and expansion of tumour cells. Vascular endothelial growth factor (VEGF) is the main endogenous pro-angiogenic factor in humans. The aim was to provide insight into the anti-VEGF activity of bioactive compounds derived from aromatic amino acids (serotonin, melatonin, 3-indoleacetic acid, 5-hydroxytryptophol and hydroxytyrosol). Experiments involved endothelial cell migration (wound-healing assay), the molecular mechanisms (ELISA assay) and the downstream effects (phospholipase C gamma 1 (PLCγ1), protein kinase B (Akt) and endothelial nitric oxide synthase (eNOS) by Western blot) on human umbilical vein endothelial cells (HUVECs). The data suggest for the first time that hydroxytyrosol interacts with surface components of the endothelial cell membrane (, preventing VEGF from activating its receptor. Serotonin and 5-hydroxytryptophol significantly inhibited HUVEC migration (98% and 50%, respectively) following the same mechanism. Conversely to other bioactive compounds, the anti-angiogenic effect of melatonin, serotonin, 3-indoleacetic acid and 5-hydroxytryptophol is not mediated via PLCγ1. However, hydroxytyrosol inhibits PLCγ1 phosphorylation. Additionally, melatonin and serotonin maintained eNOS phosphorylation and hydroxytyrosol significantly activated eNOS-all via Akt. These data provide new evidence supporting the interest in melatonin, serotonin, 3-indoleacetic acid, 5-hydroxytryptophol and hydroxytyrosol for their further exploitation as anti-VEGF ingredients in food
mTORC1 activation requires DRAM-1 by facilitating lysosomal amino acid efflux
No full textSensing nutrient availability is essential for appropriate cellular growth, and mTORC1 is a major regulator
of this process. Mechanisms causing mTORC1 activation are, however, complex and diverse. We report here an additional important step in the activation of mTORC1, which regulates the efflux of amino acids from lysosomes into the cytoplasm. This process requires DRAM-1, which binds the membrane carrier protein SCAMP3 and the amino acid transporters SLC1A5 and LAT1, directing them to lysosomes and permitting efficient mTORC1 activation.
Consequently, we show that loss of DRAM-1 also impacts pathways regulated by mTORC1, including insulin signaling, glycemic balance, and adipocyte differentiation. Interestingly, although DRAM-1 can promote autophagy, this effect on mTORC1 is autophagy independent, and autophagy only becomes important for mTORC1 activation when DRAM-1 is deleted. These findings provide important insights into mTORC1 activation and highlight the importance of DRAM-1 in growth control, metabolic homeostasis, and differentiatio