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Here, we report work on developing an enzymatic process to improve the functionalities of industrial lignin. A kraft lignin sample prepared from marine pine was treated with the high-redox-potential laccase from the basidiomycete fungus Pycnoporus cinnabarinus at three different concentrations and pH conditions, and with and without the chemical mediator 1-hydroxybenzotriazole (HBT). Laccase activity was tested in the presence and absence of kraft lignin. The optimum pH of Pci Lac was initially 4.0 in the presence and absence of lignin, but at incubation times over 6 h, higher activities were found at pH 4.5 in the presence of lignin. Structural changes in lignin were investigated by Fourier-transform infrared spectroscopy (FTIR) with differential scanning calorimetry (DSC), and solvent-extractable fractions were analyzed using high-performance size-exclusion chromatography (HPSEC) and gas chromatography-mass spectrometry (GC-MS). The FTIR spectral data were analyzed with two successive multivariate series using principal component analysis (PCA) and ANOVA statistical analysis to identify the best conditions for the largest range of chemical modifications. DSC combined with modulated DSC (MDSC) revealed that the greatest effect on glass transition temperature (Tg) was obtained at 130 U g cm -1 and pH 4.5, with the laccase alone or combined with HBT. HPSEC data suggested that the laccase treatments led to concomitant phenomena of oligomerization and depolymerization, and GC-MS revealed that the reactivity of the extractable phenolic monomers depended on the conditions tested. This study demonstrates that P. cinnabarinus laccase can be used to modify marine pine kraft lignin, and that the set of analytical methods implemented here provides a valuable tool for screening enzymatic treatment conditions., Competing Interests: The authors declare that they have no conflict of interest.

Unravelling the atomic scale diffusion that can occur at the surface, at the interface or into the bulk is challenging: multi-scale modelling approaches usually require intensive prospective calculations and moreover huge human investment. In this article, the Static Mode (SM) approach is coupled with Quantum Mechanics (QM) calculations in order to guide the exploration of the energy landscape, by optimizing the choice of events that are significant for the evolution of the system. SM enable the determination of the strain field of a set of atoms submitted to external and localized stresses, like atomic displacements. Here, we present a workflow based on the systematic SM exploration, with the objective to reduce both exploration time and human load when used with ab initio level calculations. The QMSM coupling allows the screening, scoring and selection of relevant directions that are further used to initiate and study diffusion in atomic systems. The most relevant deformations are then refined and relaxed with DFT calculations. In this paper, the overall QMSM approach is described and we discuss its use for the identification of atomic diffusion in two different systems of interest: grafting of a molecule on an oxide surface and studying the dynamical behavior of a point-defect in a bulk crystalline material.

A new biobased material based on an original strategy using lignin model compounds as natural grafting additive on a nanocellulose surface through in situ polymerization of coniferyl alcohol by the Fenton reaction at two pH values was investigated. The structural and morphological properties of the materials at the nanoscale were characterized by a combination of analytical methods, including Fourier transform infrared spectroscopy, liquid chromatography combined with mass spectrometry, nuclear molecular resonance spectroscopy, electron paramagnetic resonance spectroscopy, water sorption capacity by dynamic vapor sorption, and atomic force microscopy (topography and indentation modulus measurements). Finally, the usage properties, such as antioxidant properties, were evaluated in solution and the nanostructured casted films by radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH • ) scavenging tests. We demonstrate the structure-function relationships of these advanced CNC-lignin films and describe their dual functionalities and characteristics, namely, their antioxidant properties and the presence of persistent phenoxy radicals within the material.

Lignocellulosic biomass is receiving growing interest as a renewable source of biofuels, chemicals and materials. Lignocellulosic polymers and cellulose nanocrystals (CNCs) present high added-value potential in the nanocomposite field, but some issues have to be solved before large-scale applications. Among them, the interaction between polymers at the nanoscale and the effect of the external parameters on the mechanical properties have to be more precisely investigated. The present study aims at evaluating how the relative humidity affects the reduced Young's modulus of lignocellulosic films prepared with crystalline cellulose, glucomannan, xylan and lignin and how relative humidity changes their nanoscale adhesion properties with CNCs. Using atomic force microscopy and force volume experiments with CNC-functionalized levers, increasing the relative humidity is shown to decrease the Young's modulus values of the different films and promote their adhesion forces with CNCs. In particular, CNCs more strongly interact with glucomannan and lignin than xylan, and in the case of lignin, the oxidation of the film promotes strong variations in the adhesion force. Such results allow to better understand the lignocellulosic film properties at the nanoscale, which should lead to an improvement in the production of new highly added-value composites., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interests., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A grass soda technical lignin (PB1000) underwent a process combining solvent fractionation and treatment with an ionic liquid (IL), and a comprehensive investigation of the structural modifications was performed by using high-performance size-exclusion chromatography, 31 P NMR spectroscopy, thioacidolysis, and GC-MS. Three fractions with distinct reactivity were recovered from successive ethyl acetate (EA), butanone, and methanol extractions. In parallel, a fraction deprived of EA extractives was obtained. The samples were treated with methyl imidazolium bromide ([HMIM]Br) by using either conventional heating or microwave irradiation. The treatment allowed us to solubilize 28 % of the EA-insoluble fraction and yielded additional free phenols in all the fractions, as a consequence of depolymerization and demethylation. The gain of the combined process in terms of antioxidant properties was demonstrated through 2,2-diphenyl-1-picrylhydrazyl (DPPH . ) radical-scavenging tests. Integrating further IL safety-related data and environmental considerations, this study paves the way for the sustainable production of phenolic oligomers competing with commercial antioxidants., (© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.)

Lignocellulosic biomass is considered as a sustainable source of energy and chemicals, but its recalcitrance to bioconversion still limits its use. In this paper, a strategy based on two aspects was developed to improve our knowledge on the lignin recalcitrance to enzymatic hydrolysis. First, lignocellulosic films of cellulose nanofibrils (CNFs) with increasing content of lignin (up to 40%) were prepared. Thanks to in situ real time Atomic Force Microscopy (AFM) measurements during the hydrolysis and by comparison with biochemical assays, the use of such films allows to fully assess the importance of the lignin content and of the arrangement between CNFs and lignin on the hydrolysis efficiency. In a second time, contrary to other studies by AFM which only followed a specific structure during enzymatic processes mostly on simple systems (CNFs or cellulose nanocrystals), a quantitative analysis of in-situ time-lapse measurements was developed. It enables to accurately address lignocellulosic biomass recalcitrance mechanisms mediated by lignin at nanoscale. Such analysis could pave the way for the use of a quantitative criteria to visualize in situ deconstruction of complex lignocellulosic substrates. Coupling the use of lignocellulosic films and dynamical AFM quantitative analysis to follow the evolution of the structure at nanoscale might lead to an effective targeting of new promising bioconversion strategies.

Atomic force microscopy (AFM) experiments with functionalized tips are currently one of the most powerful tools to locally measure adhesion forces via single-molecule force spectroscopy (SMFS) measurements. The main difficulty is to precisely control the attachment of biomolecules to the cantilever. Different chemistry procedures have been developed including the use of spacer molecules. Even if a process works well for small biomolecules such as antibodies, issues remain regarding nanoparticles or larger objects such as cellulose nanocrystals because it is difficult to precisely control their coverage and homogeneity. In this work, an original procedure based on the Langmuir-Blodgett (LB) technique was implemented for lever functionalization with cellulose nanocrystals and compared with classical chemical strategies. LB shows to be almost 6.0-fold more efficient than chemical procedure in terms of cellulose nanocrystals coverage attachment. Moreover, the LB technology provides advantage of not requiring linker molecules, which could have detrimental effects such as overestimation of the interaction force. The structural characterization and SMFS measurements of lignocellulosic polymers show that this strategy enables the precise control of the lever coverage, which improves the accuracy of the adhesion measurements. Such methodology is expected to strongly impact the AFM tip/tipless functionalization and SMFS measurements in different fields.

Lignocellulosic biomass bioconversion is hampered by the structural and chemical complexity of the network created by cellulose, hemicellulose and lignin. Biological conversion of lignocellulose involves synergistic action of a large array of enzymes including the recently discovered lytic polysaccharide monooxygenases (LPMOs) that perform oxidative cleavage of cellulose. Using in situ imaging by synchrotron UV fluorescence, we have shown that the addition of AA9 LPMO (from Podospora anserina) to cellulases cocktail improves the progression of enzymes in delignified Miscanthus x giganteus as observed at tissular levels. In situ chemical monitoring of cell wall modifications performed by synchrotron infrared spectroscopy during enzymatic hydrolysis demonstrated that the boosting effect of the AA9 LPMO was dependent on the cellular type indicating contrasted recalcitrance levels in plant tissues. Our study provides a useful strategy for investigating enzyme dynamics and activity in plant cell wall to improve enzymatic cocktails aimed at expanding lignocelluloses biorefinery.

The antioxidant properties of grass lignins recovered from an alkaline industrial process and from different ethanol organosolv pretreatment processes were compared using two types of tests: (i) classical radical 2,2'-diphenyl-1-picrylhydrazyl (DPPH(•)) scavenging tests in dioxane/water or ethanol and (ii) tests involving multiphasic systems (lipid dispersion in water or cellulose film suspended in ethanol). These multiphasic systems were representative of food and packaging matrices in view of high-value applications. All lignins, in solution or in the film, effectively scavenged radicals. Moreover, they were competitive with a food commercial rosemary extract to protect linoleic acid against oxidation. Whereas the DPPH(•) test in dioxane was not discriminant, differences appeared between lignins when the test was performed in ethanol or with the multiphasic systems. Moreover, radical scavenging activity was preserved in the film even after its immersion in ethanol. Structural analysis of lignins revealed that low-molar-mass phenolics, namely p-hydroxycinnamic acids and lignin depolymerization products, governed lignin antioxidant properties in the multiphasic systems.

Novel nanocomposite coatings composed of cellulose nanocrystals (CNCs) and lignin (either synthetic or fractionated from spruce and corn stalks) were prepared without chemical modification or functionalization (via covalent attachment) of one of the two biopolymers. The spectroscopic properties of these coatings were investigated by UV-visible spectrophotometry and spectroscopic ellipsometry. When using the appropriate weight ratio of CNC/lignin (R), these nanocomposite systems exhibited high-performance optical properties, high transmittance in the visible spectrum, and high blocking in the UV spectrum. Atomic force microscopy analysis demonstrated that these coatings were smooth and homogeneous, with visible dispersed lignin nodules in a cellulosic matrix. It was also demonstrated that the introduction of nanoparticles into the medium increases the weight ratio and the CNC-specific surface area, which allows better dispersion of the lignin molecules throughout the solid film. Consequently, the larger molecular expansion of these aromatic polymers on the surface of the cellulosic nanoparticles dislocates the π-π aromatic aggregates, which increases the extinction coefficient and decreases the transmittance in the UV region. These nanocomposite coatings were optically transparent at visible wavelengths.

Interfacial affinity between lignin model compound (dehydrogenation polymer [DHP]) and cellulose nanocristals (CN) was studied before building a nanocomposite cellulose/lignin in multilayer form by spin-coating method. The adsorption isotherm of DHP was measured by ellipsometry at the liquid/CN film interface and showed that the surface concentration of adsorbed DHP increases with the bulk concentration in solution. The DHP appeared as globular structures on cellulosic film, as observed by AFM. Spreading a dense lignin layer on CN film gave rise to the disappearance of the InfraRed resonance bands related to the DHP aromatics. The film obtained from alternate layers of cellulose/DHP was transparent in visible light and had weak absorption in UV wavelengths. Optical properties measured in the visible wavelength range by ellipsometry and spectrophotometry indicated that beyond six bilayers (cellulose/DHP), the composite exhibits antireflexion properties., (Copyright © 2011 Académie des sciences. Published by Elsevier SAS. All rights reserved.)

The goal of this work is the preparation of monolayers of cellulose I nanocrystals providing flat crystalline cellulose surfaces. Suspensions of cellulose nanocrystals were prepared by hydrolyzing ramie and tunicin fibers with sulfuric acid. Due to surface grafted sulfate groups, the negatively charged, rod-like cellulose nanocrystals were found to form stable layers at the air-water interface in the presence of a cationic amphiphilic molecule such as dioctadecyldimethylammonium (DODA) used in this work. These layers were formed at different cellulose-DODA weight ratios, compressed and analyzed by tensiometry, ellipsometry and Brewster angle microscopy. At low cellulose concentrations the layers are discontinuous, becoming dense and homogeneous upon reaching a critical weight ratio, which depends on the aspect ratio of the cellulose nanocrystals. After transfer onto silicon wafers, the surface composition and morphology as well as the thickness of the films were examined by X-ray photoelectron spectroscopy, ellipsometry and atomic force microscopy. The results indicate that they are monolayer films, well structured, relatively smooth and pure. These films offer a crystalline and easily reproducible model cellulose surface.

This study analyzes the effects of some important factors of champagne technology on the ellipticity and Brewster angle microscopy (BAM) of the air/champagne interface in view of using the optical properties of the adsorption layer of base wine to forecast the stability of the champagne bubble collar. Using standard, ultrafiltered, and ultraconcentrated wines it was observed that champagne can lose amphiphilic macromolecules which adsorb on the inner glass wall of the bottle during storage, particles such as dead yeasts can adhere to the adsorption layer, a weak increase of the ethanol content during bottle fermentation can reduce significantly the ellipticity of the adsorption layer, and CO2 has no significant effect on the properties of that layer. Surprisingly, no visible differences of the adsorption layer were noticed between the experimental champagnes of the 2004 vintage of three vine varieties (Chardonnay, Pinot noir, and Pinot meunier). From analysis of all samples it is proposed that the mean value and standard deviation of the ellipticity measured during 30 min after pouring the wine in a Petri dish are physical quantities which satisfactorily characterize the adsorption layer of champagne. When needed, further characterization of the adsorption layer may be obtained by a detailed analysis of the kinetics of ellipticity during the same period and inspection of the BAM images of the interface.

4-chloro-3-[(3R)-(+)-5-chloro-1-(2,4-dimethoxybenzyl)-3-methyl-2-oxo-2,3-dihydro-1H-indol-3-yl]-N-ethyl-N-(3-pyridylmethyl)benzamide, hydrochloride (SSR126768A), a new potent and selective, orally active oxytocin (OT) receptor antagonist was characterized in several biochemical and pharmacological models. In binding studies, SSR126768A showed nanomolar affinity for rat and human recombinant and native OT receptors (K(i) = 0.44 nM) and exhibited much lower affinity for V(1a), V(1b), and V(2) receptors. In addition, it did not interact with a large number of other receptors, enzymes, and ion channels (1 microM). In autoradiographic experiments performed on at-term human pregnant uterus sections, SSR126768A dose dependently displaced [I(125)]d(CH(2))(5)[Tyr(Me)(2), Thr(4), Orn(8) (125)I-Tyr-NH(2)(9)]VT in situ labeling to OT receptors highly expressed in these tissues. In functional studies, SSR126768A behaved as a full antagonist and potently antagonized OT-induced intracellular Ca(2+) increase (K(i) = 0.50 nM) and prostaglandin release (K(i) = 0.45 nM) in human uterine smooth muscle cells. In rat isolated myometrium, OT-induced uterine contractions were competitively antagonized by SSR126768A (pA(2) = 8.47). Similarly, in human pregnant myometrial strips, SSR126768A inhibited the contractile uterine response to OT. In conscious telemetrated rats, oral administration of SSR126768A (1-10 mg/kg) produced a competitive inhibition of the dose response to OT on uterine contractions up to 24 h at 3 mg/kg p.o.; no tachyphylaxis was observed after 4-day repeated treatment. Finally, SSR126768A (30 mg/kg p.o.) significantly delayed parturition in pregnant rats in labor similar to ritodrine (10 mg/kg p.o.). Thus, SSR126768A is a potent, highly selective, orally active OT receptor antagonist with a long duration of action. This molecule could find therapeutic application as a tocolytic agent for acute and chronic oral management of preterm labor.

The author analyzes the phenomenological experience of drug in 48 drug addicts with haschisch or heroïne. The choice of the drug is linked to the severity and the type of suffering and the relationship between the drug addict and the drug is not necessarily self-destructive.