Aznag (bassin d'Ouarzazate, Maroc), nouvelle localité à sélaciens et mammifères de l'Eocène moyen (Lutétien) d'Afrique

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(EN) LIPID BASED NANOCARRIER COMPOSITIONS LOADED WITH METAL NANOPARTICLES AND THERAPEUTIC AGENT (FR) COMPOSITIONS DE NANOVECTEURS À BASE DE LIPIDES CHARGÉES DE NANOPARTICULES MÉTALLIQUES ET D'UN AGENT THÉRAPEUTIQUE

(EN)The invention relates to non-polymeric lipid-based nanocarrier compositions loaded with metal nanoparticles and at least one therapeutic agent, useful.as agents for transportation, vectorization, cellular delivery cellular targeting or cellular localization of at least one therapeutic agent. (FR)L'invention se rapporte à des compositions de nanovecteurs non polymères à base de lipides, chargées de nanoparticules métalliques et d'au moins un agent thérapeutique, utiles en tant qu'agents pour le transport, la vectorisation, l'administration cellulaire, le ciblage cellulaire ou la localisation cellulaire d'au moins un agent thérapeutique

Production and purification of recombinant human hepcidin-25 with authentic N and C-termini

Hepcidin was first identified as an antimicrobial peptide present in human serum and urine. It was later demonstrated that hepcidin is the long-sought hormone that regulates iron homeostasis in mammals. Recombinant human Hepcidin-25 (Hepc25) was expressed in Pichia pastoris using a modified version of the pPICZ alpha A vector. Hepc25 was then purified by a simple two-step chromatographic process to obtain 1.9 mg of soluble recombinant human Hepc25 per liter of culture at 96% purity. The sequence of Hepc25 and the presence of four disulfide bridges were confirmed by mass spectrometry analyses, and the recombinant Hepc25 exhibited antibacterial activity. This protocol of production and purification is the first step toward the production of human Hepc25 at a greater scale. (C) 2015 Elsevier B.V. All rights reserved

Multimodal molecular imaging of atherosclerosis: Nanoparticles functionalized with scFv fragments of an anti-αIIbβ3 antibody

Due to the wealth of actors involved in the development of atherosclerosis, molecular imaging based on the targeting of specific markers would substantiate the diagnosis of life-threatening atheroma plaques. To this end, TEG4 antibody is a promising candidate targeting the activated platelets (integrin αIIbβ3) highly represented within the plaque. In this study, scFv antibody fragments were used to functionalize multimodal imaging nanoparticles. This grafting was performed in a regio-selective way to preserve TEG4 activity and the avidity of the nanoparticles was studied with respect to the number of grafted antibodies. Subsequently, taking advantage of the nanoparticle bimodality, both near infrared fluorescence and magnetic resonance imaging of the atheroma plaque were performed in the ApoE−/− mouse model. Here we describe the design of the targeted nanoparticles, and a quantification method for their detection in mice, both ex vivo and in vivo, highlighting their value as a potential diagnosis agent.Développment d'une infrastructure française distribuée coordonné

The Trehalose Pathway Regulates Mitochondrial Respiratory Chain Content through Hexokinase 2 and cAMP in Saccharomyces cerevisiae*

In yeast, trehalose is synthesized by a multimeric enzymatic complex: TPS1 encodes trehalose 6-phosphate synthase, which belongs to a complex that is composed of at least three other subunits, including trehalose 6-phosphate phosphatase Tps2 and the redundant regulatory subunits Tps3 and Tsl1. The product of the TPS1 gene plays an essential role in the control of the glycolytic pathway by restricting the influx of glucose into glycolysis. In this paper, we investigated whether the trehalose synthesis pathway could be involved in the control of the other energy-generating pathway: oxidative phosphorylation. We show that the different mutants of the trehalose synthesis pathway (tps1Δ, tps2Δ, and tps1,2Δ) exhibit modulation in the amount of respiratory chains, in terms of cytochrome content and maximal respiratory activity. Furthermore, these variations in mitochondrial enzymatic content are positively linked to the intracellular concentration in cAMP that is modulated by Tps1p through hexokinase2. This is the first time that a pathway involved in sugar storage, i.e. trehalose, is shown to regulate the mitochondrial enzymatic content

Comparison of the immunoreactivity of TEG4-2c scFv to atherosclerotic tissues of different species by IHC analysis and ELISA assays.

<p><b>A</b> (<b>a-r)</b>: IHC assays on atherosclerotic tissues: similarly to positive controls e.g; anti-CD41 (anti-αIIb) (e), RAM11 and PGM1 (anti-CD68 antibodies targeting rabbit and human macrophages respectively) (k, q) and AP2 (anti-αIIbβ3 antibody) (a; g; m), TEG4-2c scFv specifically recognizes the injured areas of the aorta sections from different species (c; i; o). Binding of antibodies was visualized via HRP-anti-6His (scFv); HRP anti-rabbit IgG (CD41) and HRP anti-mouse IgG (RAM11, AP2). Negative controls were secondary antibody only (b; d; f; h; j; l; n; p; r). Nuclei were counterstained with hematoxylin <b>B</b>: ELISA tests on atheromatous and healthy aorta proteins: TEG4-2c shows a better recognition of atheromatous proteins. RAM11 and AP2 were used as positive controls. Negative controls were secondary antibody only. Binding of antibodies was visualized via HRP-6His or HRP-anti-mouse IgG. OD value represents absorbance at 450 nm. Values represent mean (n = 3) ± SD (error bars materialized the SD)</p

IMAC Purification of scFv TEG4-2c produced and secreted by <i>P</i>. <i>pastoris</i>.

<p>Data are standardized for 1 L culture media; the values are the mean of 7 independent experiments ± SD values.</p

A Recombinant Human Anti-Platelet scFv Antibody Produced in <i>Pichia pastoris</i> for Atheroma Targeting - Fig 3

<p><b>Purification of recombinant TEG4-2c scFv A: IMAC chromatogram</b>. The HisTrap Excel resin (5 mL) was equilibrated with 50 mM Tris-HCl pH 7.5, 500 mM NaCl (buffer A at a flow rate of 3 mL/min). <i>Pichia pastoris</i> expression broth supernatant containing the TEG4-2c scFv was injected into the column. The column was then washed with buffer A until absorbance at 280nm reached the baseline. (Dot Line): The elution was carried out in two steps using 5% and 30% buffer B (50 mM Tris-HCl pH 7.5, 500 mM NaCl, 500 mM imidazole) corresponding respectively to 25 mM and 150 mM imidazole. <b>B: Electrophoretic analysis of one step IMAC purification of recombinant TEG4-2c scFv</b>. 12% SDS-PAGE stained with colloidal blue, MW: molecular weight ladder (KDa). [S]: 5x concentrated culture supernatant. [TF]: 5x concentrated flow-through. W: 25 mM imidazole washing fraction. E: 150 mM elution fraction. E_PBS: Elution fraction dialyzed against PBS.</p