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Almahli H.,University Paris - Sud | Almahli H.,University of Aleppo | Hendra F.,University Paris - Sud | Hendra F.,IPSIT Institute Paris Sud dInnovation Therapeutique | And 8 more authors.
Chirality | Year: 2011

Diastereomeric reduction of nonactivated, hindered β-keto and chiral β-iminoesters are described. The influence of a α-stereocontrolled center on the efficiency and stereoselectivity of the reduction was studied. Reaction conditions were optimized to synthesize β-hydroxy- and β-aminoesters in good yields. In the case of chiral β-iminoesters, influence of matched/mismatched diastereomeric pairs has been assessed. Copyright © 2010 Wiley-Liss, Inc.

Barrientos L.,French Institute of Health and Medical Research | Barrientos L.,UniverSud | Marin-Esteban V.,French Institute of Health and Medical Research | Marin-Esteban V.,UniverSud | And 14 more authors.
Frontiers in Immunology | Year: 2013

Netosis is a recently described neutrophil function that leads to the release of neutrophil extracellular traps (NETs) in response to various stimuli. NETs are filaments of decondensed chromatin associated with granular proteins. In addition to their role against microorganisms, NETs have been implicated in autoimmunity, thrombosis, and tissue injury. Access to a standardized source of isolated NETs is needed to better analyze the roles of NETs. The aim of this study was to develop a procedure yielding soluble, well-characterized NET preparations from fresh human neutrophils. The calcium ionophore A23187 was chosen to induce netosis, and the restriction enzyme AluI was used to prepare large NET fragments. DNA and proteins were detected by electrophoresis and specific labeling. Some NET proteins [histone 3, lactoferrin (LF)] were quantified by western blotting, and double-stranded DNA (dsDNA) was quantified by immunofluorescence. Co-existence of dsDNA and neutrophil proteins confirmed the quality of the NET preparations. Their biological activity was checked by measuring elastase (ELA) activity and bacterial killing against various strains. Interindividual differences in histone 3, LF, ELA, and dsDNA relative contents were observed in isolated NETs. However, the reproducibility of NET preparation and characterization was validated, suggesting that this interindividual variability was rather related to donor variation than to technical bias. This standardized protocol is suitable for producing, isolating, and quantifying functional NETs that could serve as a tool for studying NET effects on immune cells and tissues. © 2013 Barrientos, Marin-Esteban, de Chaisemartin, Le-Moal, Sandré, Bianchini, Nicolas, Pallardy and Chollet-Martin.

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