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KAI2 are plant α/β hydrolase receptors, which perceive smoke-derived butenolide signals (karrikins) and putative endogenous, yet unidentified phytohormones (KAI2-ligands, KLs). The number of functional KAI2 receptors varies among plant species. It has been suggested that KAI2 gene duplication and sub-functionalization plays an adaptative role for diverse environments or ligand diversification by altering the receptor responsiveness to specific KLs. Legumes represent one of the largest families of flowering plants and contain many essential agronomic crops. Prior to legume diversification, KAI2 underwent duplication, resulting in KAI2A and KAI2B. Integrating plant genetics, ligand perception and enzymatic assays, and protein crystallography, we demonstrate that Pisum sativum KAI2A and KAI2B act as receptors and enzymes with divergent ligand stereoselectivity. KAI2B has a stronger affinity than KAI2A towards the KAI2-ligand (-)-GR24 and remarkably hydrolyses a broader range of substrates including the strigolactone-like isomer (+)-GR24. We determine the crystal structures of PsKAI2B in apo and butenolide-bound states. The biochemical and structural analyses as well as recorded mass spectra of KAI2s reveal a transient intermediate on the catalytic serine and a stable adduct on the catalytic histidine, further illuminating the role of KAI2 not only as receptors but also as bona fide enzymes. Our work uncovers the stereoselectivity of ligand perception and catalysis by evolutionarily diverged KAI2 receptors in KAR/KL signaling pathways and proposes adaptive sensitivity to KAR/KL and strigolactone phytohormones by KAI2B.

Background Glioblastomas stem-like cells (GSCs) by invading the brain parenchyma, remains after resection and radiotherapy and the tumoral microenvironment become stiffer. GSC invasion is reported as stiffness sensitive and associated with altered N-glycosylation pattern. Glycocalyx thickness modulates integrins mechanosensing, but details remain elusive and glycosylation enzymes involved are unknown. Here, we studied the association between matrix stiffness modulation, GSC migration and MGAT5 induced N-glycosylation in fibrillar 3D context. Method To mimic the extracellular matrix fibrillar microenvironments, we designed 3D-ex-polyacrylonitrile nanofibers scaffolds (NFS) with adjustable stiffnesses by loading multiwall carbon nanotubes (MWCNT). GSCs neurosphere were plated on NFSs, allowing GSCs migration and MGAT5 was deleted using CRISPR-Cas9. Results We found that migration of GSCs was maximum at 166 kPa. Migration rate was correlated with cell shape, expression and maturation of focal adhesion (FA), Epithelial to Mesenchymal Transition (EMT) proteins and (β1,6) branched N-glycan binding, galectin-3. Mutation of MGAT5 in GSC inhibited N-glycans (β1–6) branching, suppressed the stiffness dependence of migration on 166 kPa NFS as well as the associated FA and EMT protein expression. Conclusion MGAT5 catalysing multibranched N-glycans is a critical regulators of stiffness induced invasion and GSCs mechanotransduction, underpinning MGAT5 as a serious target to treat cancer.

Glioblastoma multiforme (GBM) is account for 70% of all primary malignancies of the central nervous system. Median The survival of human patients after treatment is around 15 months. There are However, several biological targets which have been reported that can be pursued using ligands with varied structures to treat this disease. In our group, we have developed several ligands that target a wide range of proteins involved in anticancer effects, such as histone deacetylase (HDACs), G protein-coupled estrogen receptor 1 (GPER), estrogen receptor-beta (ERβ), and NADPH oxidase (NOX), that were screened on bidimensional (2D) and tridimensional (3D) GBM stem cells like (GSC). Our results show that some HDAC inhibitors show antiproliferative properties at 21-32 µM as well as . These results suggest that in this 3D culture, HDACs could be the most relevant targets that are modulated to induce the antiproliferative effects that require in the future further experimental studies.

Given the limited access to freshwater compared to seawater, a growing interest surrounds the direct seawater electrolysis to produce hydrogen. However, we currently lack efficient electrocatalysts to selectively perform the oxygen evolution reaction (OER) over the oxidation of the chloride ions that are the main components of seawater. In this contribution, we report an engineering strategy to synthesize heterogeneous electrocatalysts by the simultaneous formation of separate chalcogenides of nickel (NiSx, x = 0, 2/3, 8/9, and 4/3) and cobalt (CoSx, x = 0 and 8/9) onto a carbon-nitrogen-sulfur nanostructured network. Specifically, the oxidative aniline polymerization in the presence of metallic cations was combined with the calcination to regulate the separate formation of various self-supported phases in order to target the multifunctional applicability as both hydrogen evolution reaction (HER) and OER in a simulated alkaline seawater. The OER’s metric current densities of 10 and 100 mA cm−2 were achieved at the bimetallic for only 1.60 and 1.63 VRHE, respectively. This high-performance was maintained in the electrolysis with a starting voltage of 1.6 V and satisfactory stability at 100 mA over 17 h. Our findings validate a high selectivity for OER of ~100%, which outperforms the previously reported data of 87–95%.

International audience; Investigations focusing on electrical energy storage capacitors especially the dielectric ceramic capacitors for high energy storage density are attracting more and more attention in the recent years. Ceramic capacitors possess a faster charge-discharge rate and improved mechanical and thermal properties compared with other energy storage devices such as batteries. The challenge is to obtain ceramic capacitors with outstanding mechanical, thermal and storage properties over large temperature and frequencies ranges. ABO(3) as a type of perovskites showed a strong piezoelectric, dielectric, pyroelectric, and electro-optic properties useful as energy storage and environmental devices. CaCu3Ti4O12 (CCTO) perovskite with cubic lattice (Im3 symmetry) was discovered to have a colossal dielectric constant (10(4)) that is stable over a wide range of frequencies (10 Hz-1 MHz) and temperature independence (100-300 K). The origin of this high dielectric constant is not fully established, specially because it is the same for single crystal and thin films. In this review, the history of CCTO will be introduced. The synthesis and the sintering approaches, the dopant elements used as well as the applications of CCTO will be reported. In addition to dielectrical properties useful to energy storage devices; CCTO could serve as photocatalytic materials with a very good performance in visible light.

In Streptomyces, antibiotic biosynthesis is triggered in phosphate limitation that is usually correlated with energetic stress. Polyphosphates constitute an important reservoir of phosphate and energy and a better understanding of their role in the regulation of antibiotic biosynthesis is of crucial importance. We previously characterized a gene, SLI_4384/ppk, encoding a polyphosphate kinase, whose disruption greatly enhanced the weak antibiotic production of Streptomyces lividans. In the condition of energetic stress, Ppk utilizes polyP as phosphate and energy donor, to generate ATP from ADP. In this paper, we established that ppk is co-transcribed with its two downstream genes, SLI_4383, encoding a phosin called PptA possessing a CHAD domain constituting a polyphosphate binding module and SLI_4382 encoding a nudix hydrolase. The expression of the ppk/pptA/SLI_4382 operon was shown to be under the positive control of the two-component system PhoR/PhoP and thus mainly expressed in condition of phosphate limitation. However, pptA and SLI_4382 can also be transcribed alone from their own promoter. The deletion of pptA resulted into earlier and stronger actinorhodin production and lower lipid content than the disruption of ppk, whereas the deletion of SLI_4382 had no obvious phenotypical consequences. The disruption of ppk was shown to have a polar effect on the expression of pptA, suggesting that the phenotype of the ppk mutant might be linked, at least in part, to the weak expression of pptA in this strain. Interestingly, the expression of phoR/phoP and that of the genes of the pho regulon involved in phosphate supply or saving were strongly up-regulated in pptA and ppk mutants, revealing that both mutants suffer from phosphate stress. Considering the presence of a polyphosphate binding module in PptA, but absence of similarities between PptA and known exo-polyphosphatases, we proposed that PptA constitutes an accessory factor for exopolyphosphatases or general phosphatases involved in the degradation of polyphosphates into phosphate.

The breakthrough in water electrolysis technology for the sustainable production of H2, considered as a future fuel, is currently hampered by the development of tough electrocatalytic materials. We report a new strategy of fabricating conducting polymer-derived nanostructured materials to accelerate the electrocatalytic hydrogen evolution reaction (HER), oxygen evolution reaction (OER), and water splitting. Extended physical (XRD, scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX)) and electrochemical (cyclic voltammetry (CV), linear sweep voltammetry (LSV), electrochemical impedance spectroscopy (EIS)) methods were merged to precisely characterize the as-synthesized iridium and ruthenium modified polyaniline (PANI) materials and interrogate their efficiency. The presence of Ir(+III) cations during polymerization leads to the formation of Ir metal nanoparticles, while Ru(+III) induces the formation of RuO2 oxide nanoparticles by thermal treatment, they are therefore methods for the on-demand production of oxide or metal nanostructured electrocatalysts. The findings from using 0.5 M H2SO4 highlight an ultrafast electrochemical kinetic of the material PANI-Ir for HER (36 &minus, 0 = 36 mV overpotential to reach 10 mA cm&minus, 2 at 21 mV dec&minus, 1), and of PANI-Ru for OER (1.47 &minus, 1.23 = 240 mV overpotential to reach 10 mA cm&minus, 2 at 47 mV dec&minus, 1), resulting in an efficient water splitting exactly at its thermoneutral cell voltage of 1.45 V, and satisfactory durability (96 h).

International audience; Synthetic polymers are widely employed for bone tissue engineering due to their tunable physical properties and biocompatibility. Inherently, most of these polymers display poor antimicrobial properties. Infection at the site of implantation is a major cause for failure or delay in bone healing process and the development of anti-microbial polymers is highly desired. In this study, silver nanoparticles (AgNps) were synthesized in poly-caprolactone (PCL) solution by in-situ reduction and further extruded into PCL/AgNps filaments. Customized 3D structures were fabricated using the PCL/AgNps filaments through 3D printing technique. As demonstrated by scanning electron microscopy, the 3D printed scaffolds exhibited interconnected porous structures. Furthermore, X-ray photoelectron spectroscopy analysis revealed the reduction of silver ions. Transmission electron micro-scopy along with energy-dispersive X-ray spectroscopy analysis confirmed the formation of silver nanoparticles throughout the PCL matrix. In vitro enzymatic degradation studies showed that the PCL/AgNps scaffolds dis-played 80% degradation in 20 days. The scaffolds were cytocompatible, as assessed using hFOB cells and their antibacterial activity was demonstrated on Escherichia coli. Due to their interconnected porous structure, me-chanical and antibacterial properties, these cytocompatible multifunctional 3D printed PCL/AgNps scaffolds appear highly suitable for bone tissue engineering.

International audience; The development of high-performance catalytic nanomaterials is important to implement sustainable and electrochemical energy devices of alkaline fuel cells, metal–air batteries, and electrolyzers in the envisaged energy-transition scenarios. Synthesizing effective nanocatalysts as both anode materials for oxidation of glycerol (byproduct of the biodiesel industry) and cathode materials for the oxygen reduction reaction (ORR) is still a bottleneck. Herein, we report palladium-based nanomaterials whose physicochemical and electrochemical properties are tuned by the judicious choice of the support (rGO, Vulcan XC72R), the addition of a coelement (Fe), and the structure (alloy/core–shell). The bimetallic-based electrode shows a drastically enhanced electrocatalytic performance with a beneficial shifting of the onset potential, production of high currents, and good durability for both the ORR (kinetic current density jk = 2 mA cmPd–2 or 1 A mgPd–1) and glycerol oxidation (jp = 2.3 mA cmPd–2 or 1.11 A mgPd–1 at the peak), higher than those of commercial catalysts and existing literature values. The present results also provide new fundamental insights about the accurate measurement of the kinetic metrics of the ORR by employing a rotating-disk (-ring) electrode setup in alkaline electrolytes with metallic catalysts. Indeed, the anodic scanning of the electrode from a low potential to a higher one results in an ultrafast electrochemical kinetics with a positive shift of the half-wave potential of ΔE1/2,anodic/cathodic = 60 mV from the cathodic direction to the anodic one. The kinetic current density dramatically increases, jk,anodic = 19.0×, 6.9×, 3.4×, and 2.4× jk,cathodic at 950, 900, 870, and 850 mVRHE, respectively. The advantage of the synthesis methodology relies on the nonuse of organic molecules as capping agents and surfactants in order to produce bare (ligand-free) bimetallic PdFe electrocatalysts with a clean catalytic surface in a facile and straightforward way.

In flowering plants, the α/β hydrolase DWARF14 (D14) perceives strigolactone (SL) hormones and interacts with the F-box protein MORE AXILLARY GROWTH2 (MAX2) to regulate developmental processes. The key SL biosynthetic enzyme, CAROTENOID CLEAVAGE DEOXYGENASE8 (CCD8), is present in the mossPhyscomitrium (Physcomitrella) patens,and PpCCD8-derived compounds regulate plant extension. Based on germination assays using seeds of the parasitic plantPhelipanche ramosa,we propose that these compounds are non-canonical SLs. Perception of PpCCD8-derived compounds does not require the PpMAX2 homolog. Candidate receptors are among the 13PpKAI2LIKE-A to -Lgenes, homologous to the ancestralD14paralogKARRIKIN INSENSITIVE2 (KAI2).In Arabidopsis, AtKAI2 is the receptor for a still elusive endogenous KAI2-Ligand (KL). We show that inP. patens,among SL analogs, the (+)-GR24 enantiomer is a good mimic for PpCCD8-derived compounds, while the effects of the (-)-GR24 enantiomer, a KL mimic in flowering plants, are opposite. Interaction and binding assays of seven PpKAI2L proteins using pure enantiomers pinpoint at stereoselectivity towards (-)-GR24 for the (A-E) clade. Enzyme assays highlight strong hydrolytic activity of the PpKAI2L-H protein. Moss mutants for allPpKAI2Lgene subclades were obtained and tested for their response to both enantiomers. We show thatPpKAI2L-Ato-Egenes are not involved in PpCCD8-derived compound perception, but act in a PpMAX2-dependant pathway. In contrast, mutations inPpKAI2L-G, and-Jgenes abolish the response to (+)-GR24, suggesting that encoded proteins are receptors for PpCCD8-derived SLs.

Glioblastomas stem-like cells (GSCs) by invading the brain parenchyma escape resection and radiotherapy. GSC invasion is associated with altered N-glycosylation pattern of integrins and other transmembrane proteins resulting in changed mechanosensing but details are elusive. Because the tumour microenvironment has an increased stiffness we studied the interaction between matrix stiffness, N-glycosylation and GSC migration. To mimic the fibrillar microenvironments, we designed 3D-ex-polyacrylonitrile nanofibers scaffolds (NFS) with adjustable stiffnesses by loading multiwall carbon nanotubes (MWCNT). We found that migration of GSCs was maximum at 166 kPa. Migration rate was correlated with cell shape, expression of focal adhesion (FA), Epithelial to Mesenchymal Transition (EMT) proteins and (β1,6) branched N-glycan binding, galectin-3. Mutation of MGAT5 in GSC inhibited N-glycans (β1–6) branching, suppressed the stiffness dependence of FA and EMT protein expression as well as migration on 166kPa NFS; underpinning the role of multibranched N-glycans as a critical regulator of mechanotransduction by GSC.Significance StatementDuring pathological processes in which cell migration is involved, cells undergo important functional changes in protein glycosylation and are responsive to environmental mechanical modifications. We addressed the question of the glycosylation role in mechanotransduction regulation of glioma stem cells. We created a bio-inspired 3D nanofiber scaffold (NFS) loaded with multiwall carbon nanotubes to obtain NFS of adjustable stiffness in physiological and pathological ranges. We highlighted and described a mechanism of fine mechanotransduction leading to a nonlinear migration response regarding to 3D microenvironment stiffness values. We show the importance to develop mechano-pharmacology as new therapeutic target by demonstrating the relationship existing between environmental stiffness and multibranched N-glycans catalysed by the MGAT5 enzyme to optimize directed migration.

International audience; Tartaric acid emerges as an ecofriendly organic for the biogenic preparation of micro-and nanostructured materials. For those synthesis mainly dictated by the coordination chemistry, questions remain open about the impact of the alkalinity on the nature of the resulting material as well as the properties. On the other hand, the design of advanced catalytic materials for the direct production of hydrogen peroxide from the air is particularly needed. We report herein new synthesis, physicochemical and electrocatalytic insights from bismuth-based materials. It was found that the tight control of the alkalinity of the synthesis medium through the molar ratio R = [sodium hydroxide]/[bismuth (III) nitrate] leads to the production of a library of bismuth-based materials composed of metallic, (oxy)hydroxide, and oxide structures owing to the bismuth-tartrate complexes as intermediates. Electroanalytical studies show an oxygen-to-hydrogen peroxide selectivity in 0.1 M KOH of 77, 88 and 92% for R = 0, 12 and 30, respectively. In 1 M KOH, the selectivity is 90 and 96% for R = 12 and 30, respectively. The production rate is 69 ± 3 and 44 ± 3 mol kg −1 cm −2 during 2 h, corresponding to a faradaic efficiency of 92 ± 4 and 62 ± 5% for R = 12 and 30, respectively. Those results place the present materials among the best known oxygento-hydrogen peroxide reduction catalysts without the need for preparing alloys or specific surface modification. This work contributes in engineering novel catalytic materials for the advanced application of the on-site production of valuable chemicals.

Three-dimensional (3D) gas-diffusion electrodes (GDEs) as support for catalytic nanoparticles are important to the electrolysis and fuel cell fields to facilitate reactant transport and distribution and reach higher current densities. Hence, the assembly of nanostructured catalytic particles directly onto GDEs during the synthesis could be a powerful one-step strategy to target high performance and long-term durability thanks to the induced strong nanoparticle–support interaction. We report herein the use of bromide anions to shape the anisotropic growth of nanostructured desert-rose-like gold particles directly onto the GDE, as a freestanding GDE–Au catalyst for straightforward use in electrolysis without an additional step. We showed that the bromide ions determine the anisotropy of the particles, resulting in the growth of gold into nanoplates enclosed by (111) facets. The formation of hierarchical nanostructured three-dimensional (3D) Au at the GDE surface was explained by the nanoparticle-mediated aggregation mechanism and the preferential adsorption of Br– on the (111) facet, leaving behind dominant planar structured nuclei, which then self-assemble. Electrocatalytic tests toward the glycerol electro-oxidation demonstrated that the as-synthesized freestanding surfactant- and binder-free GDE–Au material (100 μgAu cm–2) delivered a high specific peak current density of jp = 33 mA cmAu–2, which is three and seven times higher than the tested commercial Au/C nanocatalyst at loadings of 76 and 152 μgAu cm–2, respectively. This study provides future directions for the synthesis and application of nanostructured catalysts for alcohol oxidation as an alternative to anodic oxygen evolution, which consumes the majority of the electricity input during the electrolysis.

International audience; The water splitting aims to produce H2 for fuel cells and a number of industrial processes such as the ammonia synthesis and the metals refining. There is actually a major limitation about the development of cost-effective and efficient electrocatalysts to lower the cell voltage that must be overcome to perform sequentially the overall reaction that is 2H2O → 2H2 + O2. In the present study, we combined Ni and Co as the promising alternative catalytic metals to Pt-group metals as well as N and S as the doping elements to fabricate self-supported electrocatalysts for hydrogen evolution (HER) and oxygen evolution (OER) in alkaline media. To these ends, the polymerization of the aniline into polyaniline (PANI) in the presence of Ni(II) and Co(II) followed by calcination in air resulted in the formation of heterogeneous catalytic materials self-supported onto a matrix made of C, N, S, Ni and Co. Enhanced activity and durability were recorded for the hybrid and bimetallic PANI-NiCo composite.

Photoelectrochemical cells (PEC) are promising devices for hydrogen production via sunlight energy. One of the important challenges in this area is to design photoactive electrodes able to absorb visible light. A good photoelectrochemical behavior depends on the presence of surface active sites to photogenerate current at the lower possible potential for water splitting. Recent investigations in this field are focusing on perovskite materials such as CaCu3Ti4O12 (CCTO) as visible light active electrode due its outstanding structure in which CCTO encloses in its structure a visible light absorbance component (CuO). The presence CuO on the material surface is mainly responsible for trapping electrons. Therefore, CCTO photoelectrocatalysts were prepared using different amounts of graphene oxide (GO) in this work. Physico-chemical characterizations showed that the oxidation of GO at high temperatures lead to the reduction of Ti4+ to Ti3+ and Cu2+ to Cu+, causing a segregation of copper on grain boundaries regions as revealed by X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS) and Raman characterizations. Electrochemical performance and materials resistivity are measured in dark and under visible light irradiation. CCTO prepared using 3% of GO showed 50% higher photocurrent generation and hydrogen generation rate than pure phase of CCTO.

International audience; The aim of this work was to develop a bioresorbable, biodegradable and biocompatible synthetic polymer with good mechanical properties for bone tissue engineering applications. Polylactic acid (PLA) scaffolds were generated by 3D printing using the fused deposition modelling method, and reinforced by incorporation of graphene oxide (GO). Morphological analysis by scanning electron microscopy indicated that the scaffold average pore size was between 400 and 500 μm. Topography imaging revealed a rougher surface upon GO incorporation (Sa = 5.8 μm for PLA scaffolds, and of 9.9 μm for PLA scaffolds with 0.2% GO), and contact angle measurements showed a transition from a hydrophobic surface (pure PLA scaffolds) to a hydrophilic surface after GO incorporation. PLA thermomechanical properties were enhanced by GO incorporation, as shown by the 70 °C increase of the degradation peak (thermal gravimetric analysis). However, GO incorporation did not change significantly the melting point assessed by differential scanning calorimetry. Physicochemical analyses by X-ray diffraction and Raman spectroscopy confirmed the filler presence. Tensile testing demonstrated that the mechanical properties were improved upon GO incorporation (30% increase of the Young's modulus with 0.3% GO). Cell viability, attachment, proliferation and differentiation assays using MG-63 osteosarcoma cells showed that PLA/GO scaffolds were biocompatible and that they promoted cell proliferation and mineralization more efficiently than pure PLA scaffolds. In conclusion, this new 3D printed nanocomposite is a promising scaffold with adequate mechanical properties and cytocompatibility which may allow bone formation.

International audience; Here, we produced a synthetic polymer having adequate biocompatibility, biodegradability, and bioresorbability, as well as mechanical properties for applications in bone tissue engineering. We used the fused deposition modeling (FDM) based 3D printing approach in order to produce biomimetic biodegradable scaffolds made of polylactic acid (PLA). We strengthened these scaffolds by addition of exfoliated boron nitride (EBN) as filler. We demonstrated the presence of EBN by physicochemical analysis using Raman spectroscopy and X-ray diffraction (XRD). Using differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA), we found that EBN incorporation did not influence the transition temperature, but reduced the polymer crystallinity. Scanning electron microscopy for morphology evaluation showed a mean scaffold pore size of 500 μm. EBN incorporation did not affect the scaffold mechanical properties (tensile test), but modified the surface roughness. Moreover, contact angle quantification indicated that the surface of PLA/EBN scaffolds was hydrophilic and that of PLA scaffolds hydrophobic. Finally, the results of the cytotoxicity, cell attachment, and proliferation experiments using MG-63 and MC3T3 cells indicated that PLA scaffolds filled with EBN were nontoxic and compatible with osteoblastic cells and also promoted the scaffold mineralization by MG-63 cells. Altogether, our results suggest that this 3D printed nanocomposite scaffold is suitable for tissue engineering.

Bacterial lipoproteins are secreted proteins that are post-translationally lipidated. Following synthesis, preprolipoproteins are transported through the cytoplasmic membrane via the Sec or Tat translocon. As they exit the transport machinery, they are recognized by a phosphatidylglycerol::prolipoprotein diacylglyceryl transferase (Lgt), which converts them to prolipoproteins by adding a diacylglyceryl group to the sulfhydryl side chain of the invariant Cys+1residue. Lipoprotein signal peptidase (LspA or signal peptidase II) subsequently cleaves the signal peptide, liberating the α-amino group of Cys+1, which can eventually be further modified. Here, we identified thelgtandlspAgenes fromCorynebacterium glutamicumand found that they are unique but not essential. We found that Lgt is necessary for the acylation and membrane anchoring of two model lipoproteins expressed in this species: MusE, aC. glutamicummaltose-binding lipoprotein, and LppX, aMycobacterium tuberculosislipoprotein. However, Lgt is not required for these proteins’ signal peptide cleavage, or for LppX glycosylation. Taken together, these data show that inC. glutamicumthe association of some lipoproteins with membranes through the covalent attachment of a lipid moiety is not essential for further post-translational modification.

anais.laleve@inrae.frgenevieve.dujardin@i2bc.paris-saclay.fr; International audience; Artemisinin and its derivatives kill malaria parasites and inhibit the proliferation of cancer cells. In both processes, heme was shown to play a key role in artemisinin bioactivation. We found that artemisinin and clinical artemisinin derivatives are able to compensate for a mutation in the yeast Bcs1 protein, a key chaperon involved in biogenesis of the mitochondrial respiratory complex III. The equivalent Bcs1 variant causes an encephalopathy in human by affecting complex III assembly. We show that artemisinin derivatives decrease the content of mitochondrial cytochromes and disturb the maturation of the complex III cytochrome c1. This last effect is likely responsible for the compensation by decreasing the detrimental over-accumulation of the inactive pre-complex III observed in the bcs1 mutant. We further show that a fluorescent dihydroartemisinin probe rapidly accumulates in the mitochondrial network and targets cytochromes c and c1 in yeast, human cells and isolated mitochondria. In vitro this probe interacts with purified cytochrome c only under reducing conditions and we detected cytochrome c-dihydroartemisinin covalent adducts by mass spectrometry analyses. We propose that reduced mitochondrial c-type cytochromes act as both targets and mediators of artemisinin bioactivation in yeast and human cells.

Polyaniline (PANI) is a candidate for electrocatalysis, and can be combined with metal nanoparticles to fabricate high-performance electrodes for electrochemical energy conversion and storage. However, its intrinsic properties appear to be dependent on the synthesis conditions so that from the majority of the reports, it is quite difficult to establish an overall performance trend. In this contribution, we report an extensive and systematic physicochemical and electrochemical screening of the potentiality of chemically synthesized PANI as an electrode material to provide an overall understanding of the effect of the entire synthesis conditions. We have integrated different methods (TGA-DSC, XRD, SEM, EDX, FTIR, BET, CHNS, XPS, CV, and EIS) to deeply examine the as-synthesized materials and interrogate their electrocatalytic efficiency towards hydrogen evolution reaction, which was chosen as a model reaction of critical importance for H2 production from water splitting. It was found that all the synthesis parameters affect strongly the physical and electrochemical characteristics of the PANI-based materials. Specifically, XPS analysis contributed to identify the oxidation levels of the PANI samples on the basis of oxidizing agents. The outcomes provided by the study delineate a rational pathway for the further design and fabrication of PANI-based metal nanoparticles as advanced electrode materials.

Often, solid matter is separated from particle-laden flow streams using electrospun filters due to their high specific surface area, good ability to capture aerial particulate matter, and low material costs. Moreover, electrospinning allows incorporating nanoparticles to improve the filter&rsquo, s air filtration efficiency and bacterial removal. Therefore, a new, improved polyacrylonitrile (PAN) nanofibers membrane that could be used to remove air pollutants and also with antibacterial activity was developed. We engineered three different filters that are characterized by the different particles embedded in the PAN nanofibers: titanium dioxide (TiO2), zinc oxide (ZnO), and silver (Ag). Then, their filtration performance was assessed by quantifying the filtration of sodium chloride (NaCl) aerosol particles of 9 to 300 nm in diameter using a scanning mobility particle sizer. The TiO2_F filter displayed the smallest fiber diameter and the highest filtration efficiency (&asymp, 100%). Conversely, the Ag_F filter showed the highest quality factor (&asymp, 0.06 Pa&minus, 1) because of the lower air pressure drop. The resulting Ag_F nanofibers displayed a very good antibacterial activity using an Escherichia coli suspension (108 CFU/mL). Moreover, the quality factor of these membranes was higher than that of the commercially available nanofiber membrane for air filtration.

International audience; Gelatin is a fascinating biomaterial due to its biocompatibility, biodegradability and chemical composition. The engineering of gelatin scaffolds with the most appropriate architecture is crucial for tissue engineering applications. Currently available porogen assisted scaffold fabrication technique are limited for the fabrication of repeatable and customized scaffolds. Three dimensional printing is an appropriate technique to prepare customized structure. In this report, customized gelatin scaffolds were fabricated using 3D printed polyvinyl alcohol (PVA) sacrificial molds. Gelatin was stabilized through crosslinking and the PVA was solubilized to obtain 3D customized gelatin scaffolds. The gelatin scaffolds displayed tunable pore size from 350 to 1200 ​μm. The influence of PVA infill density on the gelatin scaffold porous structure and interconnectivity was also analyzed. Furthermore, the gelatin scaffold stability in phosphate buffered saline solution and their swelling capacity demonstrated that they were effectively crosslinked. The scaffold cytocompatibility and the good cell attachment and proliferation on these scaffolds showed that the sacrificial mold-assisted 3D printing is suitable for producing customizable gelatin scaffolds for biomedical applications.

International audience; Gelatin is a biodegradable biopolymer obtained by collagen denaturation, which shows poor mechanical properties. Hence, improving its mechanical properties is very essential towards the fabrication of efficient nontoxic material for biomedical applications. For this aim, various methods are employed using external fillers such as ceramics or bioglass. In this report, we introduce boron nitride (BN) reinforced gelatin as a new class of two dimensional biocompatible nanomaterials. The effect of the nanofiller on the mechanical behavior isanalyzed. BN is efficiently exfoliated using the biopolymer gelatin as shown through Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD). The exfoliated BN reinforces gelatin electrospun fibers, which results in an increase in the Young’s modulus. The ESM are stable after the glutaraldehyde cross-linking and the fibrous morphology is preserved. The cross-linked gelatin/BN ESM is highly bioactive in forming bone like hydroxyapatite as shown by scanning electron microscopy. Due to their enhanced mineralization ability, the cross-linked ESM have been tested on human bone cells (HOS osteosarcoma cell line). The cell attachment, proliferation and biocompatibility results show that the ESM are nontoxic and biodegradable. The analysis of osteoblast gene expression and the measurement of alkaline phosphatase activity confirm that these materials are suitable for bone tissue engineering

Ubiquitous electrochemistry is expected to play a major role for reliable energy supply as well as for production of sustainable fuels and chemicals. The fundamental understanding of organics-based electrocatalysis in alkaline media at the solid-liquid interface involves complex mechanisms and performance descriptors (from the electrolyte and reaction intermediates), which undermine the roads towards advance and breakthroughs. Here, we review and diagnose recently designed strategies for the electrochemical conversion of organics into electricity and/or higher-value chemicals. To tune the mysterious workings of nanocatalysts in electrochemical devices, we examine the guiding principles by which the performance of a particular electrode material is governed, thus highlighting various tactics for the development of synthesis methods for nanomaterials with specific properties. We end by examining the production of chemicals by using electrochemical methods, from selective oxidation to reduction reactions. The intricate relationship between electrode and selectivity encourages both of the communities of electrocatalysis and organic synthesis to move forward together toward the renaissance of electrosynthesis methods.

The study reports on the preparation of polyacrylonitrile fiber paper (PANFP) functionalized with polydopamine (PD) and silver nanoparticles (Ag NPs), named as Ag NPs/PD/PANFP. The composite material was obtained via a simple two-step chemical process. First, a thin polydopamine layer was coated onto the PANFP surface through immersion into an alkaline dopamine (pH 8.5) aqueous solution at room temperature. The reductive properties of polydopamine were further exploited for the deposition of Ag NPs. The morphology and chemical composition of the composite material were characterized using scanning electron microscopy (SEM), X-ray diffraction pattern (XRD) and X-ray photoelectron spectroscopy (XPS). The catalytic activity of the nanocomposite was evaluated for the reduction of 4-nitrophenol using sodium borohydride (NaBH 4 ) at room temperature. The Ag NPs/PD/PANFP displayed good catalytic performance with a full reduction of 4-nitrophenol into the corresponding 4-aminophenol within 30 min. Moreover, the composite material exhibited a good stability up to 4 cycles without a significant loss of its catalytic activity.

Ubiquinone (UQ), also referred to as coenzyme Q, is a widespread lipophilic molecule in both prokaryotes and eukaryotes in which it primarily acts as an electron carrier. Eleven proteins are known to participate in UQ biosynthesis in Escherichia coli, and we recently demonstrated that UQ biosynthesis requires additional, nonenzymatic factors, some of which are still unknown. Here, we report on the identification of a bacterial gene, yqiC, which is required for efficient UQ biosynthesis, and which we have renamed ubiK. Using several methods, we demonstrated that the UbiK protein forms a complex with the C-terminal part of UbiJ, another UQ biogenesis factor we previously identified. We found that both proteins are likely to contribute to global UQ biosynthesis rather than to a specific biosynthetic step, because both ubiK and ubiJ mutants accumulated octaprenylphenol, an early intermediate of the UQ biosynthetic pathway. Interestingly, we found that both proteins are dispensable for UQ biosynthesis under anaerobiosis, even though they were expressed in the absence of oxygen. We also provide evidence that the UbiK-UbiJ complex interacts with palmitoleic acid, a major lipid in E. coli. Last, in Salmonella enterica, ubiK was required for proliferation in macrophages and virulence in mice. We conclude that although the role of the UbiK-UbiJ complex remains unknown, our results support the hypothesis that UbiK is an accessory factor of Ubi enzymes and facilitates UQ biosynthesis by acting as an assembly factor, a targeting factor, or both. Agence Nationale de la Recherche ANR-15-CE11-0001-02 Centre National de la Recherche Scientifique PICS07279 French State Program "Investissements d'Avenir" ANR-11-LABX-0011

International audience

In order to colonize environments with large O2 gradients or fluctuating O2 levels, bacteria have developed metabolic responses that remain incompletely understood. Such adaptations have been recently linked to antibiotic resistance, virulence, and the capacity to develop in complex ecosystems like the microbiota. Here, we identify a novel pathway for the biosynthesis of ubiquinone, a molecule with a key role in cellular bioenergetics. We link three uncharacterized genes of Escherichia coli to this pathway and show that the pathway functions independently from O2. In contrast, the long-described pathway for ubiquinone biosynthesis requires O2 as a substrate. In fact, we find that many proteobacteria are equipped with the O2-dependent and O2-independent pathways, supporting that they are able to synthesize ubiquinone over the entire O2 range. Overall, we propose that the novel O2-independent pathway is part of the metabolic plasticity developed by proteobacteria to face various environmental O2 levels., Most bacteria can generate ATP by respiratory metabolism, in which electrons are shuttled from reduced substrates to terminal electron acceptors, via quinone molecules like ubiquinone. Dioxygen (O2) is the terminal electron acceptor of aerobic respiration and serves as a co-substrate in the biosynthesis of ubiquinone. Here, we characterize a novel, O2-independent pathway for the biosynthesis of ubiquinone. This pathway relies on three proteins, UbiT (YhbT), UbiU (YhbU), and UbiV (YhbV). UbiT contains an SCP2 lipid-binding domain and is likely an accessory factor of the biosynthetic pathway, while UbiU and UbiV (UbiU-UbiV) are involved in hydroxylation reactions and represent a novel class of O2-independent hydroxylases. We demonstrate that UbiU-UbiV form a heterodimer, wherein each protein binds a 4Fe-4S cluster via conserved cysteines that are essential for activity. The UbiT, -U, and -V proteins are found in alpha-, beta-, and gammaproteobacterial clades, including several human pathogens, supporting the widespread distribution of a previously unrecognized capacity to synthesize ubiquinone in the absence of O2. Together, the O2-dependent and O2-independent ubiquinone biosynthesis pathways contribute to optimizing bacterial metabolism over the entire O2 range.

International audience; The first non-natural derivative of the rare D-glucose-2-phosphate (G2P), namely glucose-2-(O-lactic acid phosphate) (G2LP), has been synthesized. When used as sole carbon source, G2LP enables bacterial growth of the plant pathogenic strain Agrobacterium fabrum C58 (formerly referred to as Agrobacterium tumefa-ciens). X-ray crystallography and affinity measurements investigations reveal that G2LP binds the periplasmic binding protein (PBP) AccA similarly to the natural compounds and with the same affinity. Moreover, enzy-matic assays show that it is able to serve as substrate of the phosphodiesterase AccF. The properties found for G2LP demonstrate that the very unusual glucose-2-phosphoryl residue, present in G2LP, can be used as structural feature for designing non-natural systems fully compatible with the Acc cascade of A. fabrum.

Glioblastoma Multiforme (GBM) invasiveness renders complete surgical resection impossible and highly invasive Glioblastoma Initiating Cells (GICs) are responsible for tumour recurrence. Their dissemination occurs along pre-existing fibrillary brain structures comprising the aligned myelinated fibres of the corpus callosum (CC) and the laminin (LN)-rich basal lamina of blood vessels. The extracellular matrix (ECM) of these environments regulates GIC migration, but the underlying mechanisms remain largely unknown. In order to recapitulate the composition and the topographic properties of the cerebral ECM in the migration of GICs, we have set up a new aligned polyacrylonitrile (PAN)-derived nanofiber (NF) scaffold. This system is suitable for drug screening as well as discrimination of the migration potential of different glioblastoma stem cells. Functionalisation with LN increases the spatial anisotropy of migration and modulates its mode from collective to single cell migration. Mechanistically, equally similar to what has been observed for mesenchymal migration of GBM in vivo, is the upregulation of galectin-3 and integrin-β1 in Gli4 cells migrating on our NF scaffold. Downregulation of Calpain-2 in GICs migrating in vivo along the CC and in vitro on LN-coated NF underlines a difference in the turnover of focal adhesion (FA) molecules between single-cell and collective types of migration.

International audience; Photoelectrochemical water splitting under visible light has attracted attention for renewable hydrogen production. Despite prevalent investigations, many challenges still hindered an efficient energy conversion, such as enhancing the reaction efficiency in visible light. Thus controlling the photoelectrode materials is an essential step in designing new materials for water splitting. CaCu3Ti4O12 (CCTO) has received great attention as photocatalyst under solar light due to its combined band gap as result of the presence in its structure of TiO2 active in UV light and CuO active under visible light. In this work, a cubic CCTO with different amount of exfoliated hexagonal boron nitride has been synthesized. The produced materials were fully characterized. Physico-chemical characterizations demonstrate that bore and nitrogen co-doped CCTO causes an increase in grain boundaries thickness, and thus leads to shift peaks in XRD and Raman spectra. UV–Vis diffuse reflectance spectroscopy showed a decrease of the bang gap value after adding boron nitride nanosheets into CCTO. The electrochemical performance and the resistivity of the obtained materials were performed using electrochemical impedance spectroscopy in dark and under visible light exposition. The onset potential recorded for CCTO with 3% of boron nitride nanosheets (Eg = 2.9), is around 0.5 V and the current generation is 16 times more than pure phase of CCTO. This work shows that CCTO with 3% of boron nitride nanosheets can be considered as an active photoelectrocatalyst under visible light irradiation for water splitting. Hydrogen production measurements show that the rate of H2 production reached arround 8.1 µmol/h under visible light after adding 3% of BN nanosheets to CCTO which presents 16 times increase in comparison with pure CCTO.

International audience; Human exposure to air pollution and especially to nanoparticles is increasing due to the combustion of carbon-based energy vectors. Fibrous filters are among the various types of equipment potentially able to remove particles from the air. Nanofibers are highly effective in this area; however, their utilization is still a challenge due to the lack of studies taking into account both nanoparticle collection efficiency and antibacterial effect. The aim of this work is to produce and evaluate novel silver/polyacrylonitrile (Ag/PAN) electrospun fibers deposited on a nonwoven substrate to be used as air filters to remove nanoparticles from the air and also showing antibacterial activity. In order to determine the optimum manufacturing conditions, the effects of several electrospinning process parameters were analyzed such as solution concentration, collector to needle distance, flow rate, voltage, and duration. Ag/PAN nanofibers were characterized by X-ray diffraction (XRD), Transmission Electron Microscopy (TEM), Fourier Transform Infra-Red spectroscopy (FTIR), Energy-dispersive X-ray spectroscopy (EDX), X-ray photoelectron spectroscopy (XPS), and Scanning Electron Microscopy (SEM). In addition, filtration performances were determined by measuring the pressure drop and collection efficiency of sodium chloride (NaCl) aerosol particles (9 to 300 nm diameters) using Scanning Mobility Particle Sizers (SMPS). Filters with high filtration efficiency (≈100%) and high-quality factor (≈0.05 Pa-1) were obtained even adding different concentrations of Ag nanoparticles (AgNPs) to PAN nanofibers. The resultant Ag/PAN nanofibers showed excellent antibacterial activity against 10 4 CFU/ml E.coli bacteria.

International audience; The design and fabrication of durable nanocatalysts to efficiently and selectively electro-oxidize organic molecules toward value-added product(s) is an important starting point for the future deployment of electrochemical "cogeneration" devices with the triple advantage of producing electricity, heat and fuels/chemicals. This interdisciplinary research requires a synergistic effort from three communities of electrochemistry/electrocatalysis, material science and organic chemistry. To this end, we chose to explore the integration of different electrochemical and analytical techniques to study the outstanding ability of gold-based nanomaterials in the electrocatalytic oxidation of mono- and di-saccharides. In order to rationalize the previous outcomes on the selective catalysts for glucose-to-gluconate conversion toward cogenerating organic electrosynthesis (ECS Trans., 77, 1547 (2017)), a multivariate study is carried out in alkaline and neutral pHs by examining three substrates, glucose, galactose, lactose and different electrodes. Electroanalytical investigation revealed that these substrates are selectively oxidized at their Cl-position (two transferred electrons). At pH 7.4, the corresponding lactone and acid forms were detected by their specific vibration bands of 1744 and 1780 cm(-1), which is explained by a local pH decrease within the thin-electrolyte. Our study delineates a broad strategy for organic electrosynthesis in electrochemical cells by controlling electrode materials fabrication. (C) The Author(s) 2018. Published by ECS.

Premature termination codons (PTCs) are generally associated with severe forms of genetic diseases. Readthrough of in-frame PTCs using small molecules is a promising therapeutic approach. Nonetheless, the outcome of preclinical studies has been low and variable. Treatment efficacy depends on: 1) the level of drug-induced readthrough, 2) the amount of target transcripts, and 3) the activity of the recoded protein. The aim of the present study was to identify, in the cystic fibrosis transmembrane conductance regulator (CFTR) model, recoded channels from readthrough therapy that may be enhanced using CFTR modulators. First, drug-induced readthrough of 15 PTCs was measured using a dual reporter system under basal conditions and in response to gentamicin and negamycin. Secondly, exon skipping associated with these PTCs was evaluated with a minigene system. Finally, incorporated amino acids were identified by mass spectrometry and the function of the predicted recoded CFTR channels corresponding to these 15 PTCs was measured. Nonfunctional channels were subjected to CFTR-directed ivacaftor-lumacaftor treatments. The results demonstrated that CFTR modulators increased activity of recoded channels, which could also be confirmed in cells derived from a patient. In conclusion, this work will provide a framework to adapt treatments to the patient's genotype by identifying the most efficient molecule for each PTC and the recoded channels needing co-therapies to rescue channel function., This study identified readthrough-recoded CFTR channels, the activity of which could be enhanced using CFTR modulators http://ow.ly/f7Gd30hBCeG

International audience; The synthesis of tunable boron nitride (BN) nanotubes is the subject of intense research because of their potential applications in various fields, from smart textile materials to emerging environmental technologies. However, the preparation of these nanomaterials remains extremely challenging. Herein, proof-of-concept results on the scalable synthesis of boron nitride nanotubes with tunable dimensions are reported. Combining atomic layer deposition (ALD) of boron nitride on carbon nanofibers template supports and successive annealing steps at high temperature, BN nanotubes with controllable properties are prepared. The successive annealing steps enable the crystallization of BN and the combustion of the initial carbon supports. The self-limiting nature of the ALD process allows for the scalable coating of large scale carbon fibers matrix. High-resolution transmission electron microscopy measurements demonstrate the exquisite dimensions control at the nanoscale and the presence of crystalline BN. The proof-of-concept results presented allow for independent control of the inner and external diameter dimensions, opening up unique possibilities for the precise engineering of BN nanotubes properties. The nanotubes present good mechanical properties, are easy to handle, and the assessment of their sorption properties shows that they absorb up to 110 times their own weight in oils while repelling water, which opens prospects for water purification.

International audience; Selenoproteins are essential components of antioxidant defense, redox homeostasis, and cell signaling in mammals, where selenium is found in the form of a rare amino acid, selenocysteine. Selenium, which is often limited both in food intake and cell culture media, is a strong regulator of selenoprotein expression and selenoenzyme activity. Aging is a slow, complex, and multifactorial process, resulting in a gradual and irreversible decline of various functions of the body. Several cellular aspects of organismal aging are recapitulated in the replicative senescence of cultured human diploid fibroblasts, such as embryonic lung fibroblast WI-38 cells. We previously reported that the long-term growth of young WI-38 cells with high (supplemented), moderate (control), or low (depleted) concentrations of selenium in the culture medium impacts their replicative lifespan, due to rapid changes in replicative senescence-associated markers and signaling pathways. In order to gain insight into the molecular link between selenium levels and replicative senescence, in the present work, we have applied a quantitative proteomic approach based on 2-Dimensional Differential in-Gel Electrophoresis (2D-DIGE) to the study of young and presenescent cells grown in selenium-supplemented, control, or depleted media. Applying a restrictive cut-off (spot intensity \textpm50% and a p value \textless 0.05) to the 2D-DIGE analyses revealed 81 differentially expressed protein spots, from which 123 proteins of interest were identified by mass spectrometry. We compared the changes in protein abundance for three different conditions: (i) spots varying between young and presenescent cells, (ii) spots varying in response to selenium concentration in young cells, and (iii) spots varying in response to selenium concentration in presenescent cells. Interestingly, a 72% overlap between the impact of senescence and selenium was observed in our proteomic results, demonstrating a strong interplay between selenium, selenoproteins, and replicative senescence.

International audience; Understanding the mechanisms of protein oligomerization and aggregation is a major concern for biotechnology and medical purposes. However, significant challenges remain in determining the mechanism of formation of these superstructures and the environmental factors that can precisely modulate them. Notably the role that a functional ligand plays in the process of protein aggregation is largely unexplored. We herein address these issues with an original flavin-dependent RNA methyltransferase (TrmFO) used as a protein model since this protein employs a complex set of cofactors and ligands for catalysis. Here, we show that TrmFO carries an unstable protein structure that can partially mis-unfold leading to either formation of irregular and nonfunctional soluble oligomers endowed with hyper-thermal stability or large amorphous aggregates in the presence of salts. Mutagenesis confirmed that this peculiarity is an intrinsic property of a polypeptide and it is independent of the flavin coenzyme. Structural characterization and kinetic studies identified several regions of the protein that enjoy conformational changes and more particularly pinpointed the N-terminal subdomain as being a key element in the mechanisms of oligomerization and aggregation. Only stabilization of this region via tRNA suppresses these aberrant protein states. Although protein chaperones emerged as major actors against aggregation, our study emphasizes that other powerful mechanisms exist such as the stabilizing effect of functional assemblies that provide an additional layer of protection against the instability of the proteome.

International audience; To facilitate production of functional enzymes and to study their mechanisms, especially in the complex cases of coenzyme-dependent systems, activation of an inactive apoenzyme preparation with a catalytically competent coenzyme intermediate is an attractive strategy. This is illustrated with the simple chemical synthesis of a flavin-methylene iminium compound previously proposed as a key intermediate in the catalytic cycle of several important flavoenzymes involved in nucleic acid metabolism. Reconstitution of both flavin-dependent RNA methyltransferase and thymidylate synthase apoproteins with this synthetic compound led to active enzymes for the C5-uracil methylation within their respective transfer RNA and dUMP substrate. This strategy is expected to be of general application in enzymology.

International audience; Significant reduction of the amount of precious metals in catalysts is a major challenge. We report the synthesis of high-performance carbon paper-boron nitride-palladium (CP-BN-Pd) electrocatalytic electrodes. The nanocatalysts consist of Pd nanoparticles of 5 nm supported on an ultrathin BN film prepared by atomic layer deposition (ALD), covering the microfibers of gas-diffusion electrodes (GDL). These electrodes present significantly enhanced electrocatalytic performance towards oxygen reduction (ORR) and C2 alcohols oxidation reactions and outperform the reported data for those alcohols in alkaline media, reaching a peak current of 17 amps/mg(pd) in 1 M NaOH + 1 M ethanol. The ageing tests reveal excellent stability of the electrochemically active surface area even after 1000 cycles, and the ethanol oxidation activity shows negligible decay of 1% whereas commercial Pd/C show prominent decay of 44%. The use of this heterogeneous active interface opens a new route for the development of efficient and low-metal content nanocatalysts.