Interaction of Avibactam with Class B Metallo-ß-Lactamases.

ß-Lactamases are the most important mechanisms of resistance to the ß-lactam antibacterials. There are two mechanistic classes of ß-lactamases: the serine ß-lactamases (SBLs) and the zinc-dependent metallo-ß-lactamases (MBLs). Avibactam, the first clinically useful non-ß-lactam ß-lactamase inhibitor, is a broad-spectrum SBL inhibitor, which is used in combination with a cephalosporin antibiotic (ceftazidime). There are multiple reports on the interaction of avibactam with SBLs but few such studies with MBLs. We report biochemical and biophysical studies on the binding and reactivity of avibactam with representatives from all 3 MBL subfamilies (B1, B2, and B3). Avibactam has only limited or no activity versus MBL-mediated resistance in pathogens. Avibactam does not inhibit MBLs and binds only weakly to most of the MBLs tested; in some cases, avibactam undergoes slow hydrolysis of one of its urea N-CO bonds followed by loss of CO2, in a process different from that observed with the SBLs studied. The results suggest that while the evolution of MBLs that more efficiently catalyze avibactam hydrolysis should be anticipated, pursuing the development of dual-action SBL and MBL inhibitors based on the diazabicyclooctane core of avibactam may be productive.

When Microplastic Is Not Plastic: The Ingestion of Artificial Cellulose Fibers by Macrofauna Living in Seagrass Macrophytodetritus.

Dead leaves of the Neptune grass, Posidonia oceanica (L.) Delile, in the Mediterranean coastal zone, are colonized by an abundant "detritivorous" invertebrate community that is heavily predated by fishes. This community was sampled in August 2011, November 2011, and March 2012 at two different sites in the Calvi Bay (Corsica). Ingested artificial fibers (AFs) of various sizes and colors were found in 27.6% of the digestive tracts of the nine dominant species regardless of their trophic level or taxon. No seasonal, spatial, size, or species-specific significant differences were revealed; suggesting that invertebrates ingest AFs at constant rates. Results showed that, in the gut contents of invertebrates, varying by trophic level, and across trophic levels, the overall ingestion of AFs was low (approximately 1 fiber per organism). Raman spectroscopy revealed that the ingested AFs were composed of viscose, an artificial, cellulose-based polymer. Most of these AFs also appeared to have been colored by industrial dyes. Two dyes were identified: Direct Blue 22 and Direct Red 28. The latter is known for being carcinogenic for vertebrates, potentially causing environmental problems for the P. oceanica litter community. Techniques such as Raman spectroscopy are necessary to investigate the particles composition, instead of relying on fragment size or color to identify the particles ingested by animals.

Detection of Anthropogenic Particles in Fish Stomachs: An Isolation Method Adapted to Identification by Raman Spectroscopy.

Microplastic particles (MP) contaminate oceans and affect marine organisms in several ways. Ingestion combined with food intake is generally reported. However, data interpretation often is circumvented by the difficulty to separate MP from bulk samples. Visual examination often is used as one or the only step to sort these particles. However, color, size, and shape are insufficient and often unreliable criteria. We present an extraction method based on hypochlorite digestion and isolation of MP from the membrane by sonication. The protocol is especially well adapted to a subsequent analysis by Raman spectroscopy. The method avoids fluorescence problems, allowing better identification of anthropogenic particles (AP) from stomach contents of fish by Raman spectroscopy. It was developed with commercial samples of microplastics and cotton along with stomach contents from three different Clupeiformes fishes: Clupea harengus, Sardina pilchardus, and Engraulis encrasicolus. The optimized digestion and isolation protocol showed no visible impact on microplastics and cotton particles while the Raman spectroscopic spectrum allowed the precise identification of microplastics and textile fibers. Thirty-five particles were isolated from nine fish stomach contents. Raman analysis has confirmed 11 microplastics and 13 fibers mainly made of cellulose or lignin. Some particles were not completely identified but contained artificial dyes. The novel approach developed in this manuscript should help to assess the presence, quantity, and composition of AP in planktivorous fish stomachs.

Determination of the Al2O3 content in NaF-AlF3-CaF2-Al2O3 melts at 950 °C by Raman spectroscopy.

The in situ control of the chemical composition of industrial aluminum smelter is a challenge mainly for physicochemical reasons: high temperature, high surrounding electromagnetic field, and the highly corrosive molten salt electrolyte to deal with. In previous works, we proposed that Raman spectroscopy is a method of choice that could be adapted to real smelters. The laboratory study presented here relies on reproducible Raman spectra recorded on molten mixtures whose compositions are identical to those used during the production of aluminum. A normalization procedure for the Raman spectra is proposed based on the equilibria taking place in the bath. In addition, we discuss two quantitative models to determine the alumina content from the Raman spectra of the molten NaF-AlF3-CaF2-Al2O3 electrolytes. Univariate and multivariate approaches are applied to determine both the COx (alumina content) and the CR (NaF/AlF3 molar ratio) by Raman spectroscopy without referring to an additional internal reference of intensity. The procedure was successfully tested and validated on industrial samples.

Direct determination of the NaF/AlF3 molar ratio by Raman spectroscopy in NaF-AlF3-CaF2 melts at 1000 °C.

For the last 40 years, Raman spectroscopy has been very useful in investigating the structure of corrosive molten salts, such as the cryolite-based melts widely used as electrolyte in the Hall-Heroult process. Even if this process remains the most economically efficient for metallic aluminum electro-production, it suffers from a high energy loss, which is dependent on the melt composition. Therefore, controlling the chemical composition of the electrolyte is essential. The present paper proposes to apply Raman spectroscopy for the direct determination of the NaF-AlF3 molar ratio in NaF-AlF3-CaF2-based melts. Despite the experimental difficulties, a calibration curve based on equilibria taking place in the melt has been developed and the procedure has been successfully compared to industrial samples of known compositions. The possible exportation of the laboratory scale procedure to an industrial environment application for the control of the Hall-Heroult process is finally discussed.

Online monitoring of heterogeneous polymerizations in supercritical carbon dioxide by Raman spectroscopy.

A novel setup is described to monitor the kinetics of homogeneous and heterogeneous reactions online using Raman spectroscopy under high pressure. The arrangement is based on a high-pressure reactor equipped with a sapphire window in combination with a mobile probe that allows the collection of the back-scattered Raman signal at 360°.

Anthropogenic particles in the stomach contents and liver of the freshwater fish Squalius cephalus.

Anthropogenic particles (APs) are a very broad category of particles produced directly or indirectly by human activities. Their ingestion by biota is well studied in the marine environment. In contrast, studies on AP ingestion in wild freshwater organisms are scarce despite high contamination levels in some rivers and lakes. In this study, we aimed to evaluate the ingestion of APs and the possible occurrence of APs in the liver and muscle of a freshwater fish, Squalius cephalus, from the Parisian conurbation. After isolation, the particles were analyzed using Raman spectroscopy. In sixty stomachs, eighteen APs were found, half of which were plastics and the other half were dyed particles. Twenty-five percent of sampled individuals had ingested at least one AP. The mean length of the APs was 2.41 mm. No significant difference was found between the sites upstream and downstream of Paris. Additionally, 5% of sampled livers contained one or more APs, which were characterized as microplastics (MPs). No APs were found in the muscle tissue. The majority of APs isolated from stomach contents were fibers, which is similar to the findings of a previous river contamination study. This highlights that fish could be more exposed to fibers than previously thought and that more studies on the impacts of fiber ingestion are required. Despite their low occurrence, MPs are reported, for the first time, in the liver of a wild freshwater fish species. While the pathways and impacts are still unknown, MPs also occur in liver of marine mollusks and fish. Physiological in vitro studies are needed to better evaluate the impacts of such phenomena.

Microplastics in livers of European anchovies (Engraulis encrasicolus, L.).

Microplastics (MPs) are thought to be ingested by a wide range of marine organisms before being excreted. However, several studies in marine organisms from different taxa have shown that MPs and nanoplastics could be translocated in other organs. In this study, we investigated the presence of MPs in the livers of commercial zooplanktivorous fishes collected in the field. The study focuses mainly on the European anchovy Engraulis encrasicolus but concerns also the European pilchard Sardina pilchardus and the Atlantic herring Clupea harengus. Two complementary methodologies were used to attest the occurrence of MPs in the hepatic tissue and to exclude contamination. 1) MPs were isolated by degradation of the hepatic tissue. 2) Cryosections were made on the livers and observed in polarized light microscopy. Both methods separately revealed that MPs, mainly polyethylene (PE), were translocated into the livers of the three clupeid species. In anchovy, 80 per cent of livers contained relatively large MPs that ranged from 124 µm to 438 µm, showing a high level of contamination. Two translocation pathways are hypothesized: (i) large particles found in the liver resulted from the agglomeration of smaller pieces, and/or (ii) they simply pass through the intestinal barrier. Further studies are however required to understand the exact process.

Morphology of the filtration apparatus of three planktivorous fishes and relation with ingested anthropogenic particles.

Anthropogenic particles (APs), including microplastics, are ingested by a wide variety of marine organisms. Exposure of Clupeiformes (e.g. herrings, anchovies, sardines) is poorly studied despite their economic and ecological importance. This study aims to describe the morphology of the filtration apparatus of three wild-caught Clupeiformes (Sardina pilchardus, Clupea harengus and Engraulis encrasicolus) and to relate the results to ingested APs. Consequently, the species with the more efficient filtration apparatus will be more likely to ingest APs. We hypothesized that sardines were the most exposed species. The filtration area and particle retention threshold were determined in the three species, with sardines displaying the highest filtration area and the closest gill rakers. Sardines ingested more fibers and smaller fragments, confirming that it is the most efficient filtering species. These two results lead to the conclusion that, among the three studied, the sardine is the species most exposed to APs.

Fluorinated Alcohols as Activators for the Solvent-Free Chemical Fixation of Carbon Dioxide into Epoxides.

The addition of fluorinated alcohols to onium salts provides highly efficient organocatalysts for the chemical fixation of CO2 into epoxides under mild experimental conditions. The combination of online kinetic studies, NMR titrations and DFT calculations allows understanding this synergistic effect that provides an active organocatalyst for CO2 /epoxides coupling.

Structure and Thermodynamics of NaF-AlF(3) Melts with Addition of CaF(2) and MgF(2).

The NaF-AlF(3) system with additions of CaF(2) and MgF(2) has been studied with Raman and vapor pressure measurements for 3 >/= CR (NaF/AlF(3) molar ratio) >/= 1 and up to 50 mol % additive. The results show that the binary melt can be described using the two equilibria AlF(6)(3)(-) = AlF(6)(2)(-) + F(-) and AlF(5)(2)(-) = AlF(4)(-) + F(-) with equilibrium constants 0.25 and 0.05, respectively, at 1293 K. Both reactions have positive reaction enthalpies. The first equilibrium is strongly shifted to the right resulting in a melt mixture with very low AlF(6)(3)(-) concentrations even at the Na(3)AlF(6) composition. Evidence for nonideal mixing of anions was found. For the ternaries, models based on Raman data are presented and compared with vapor pressure measurements. Good agreement is observed when association between the additives, CaF(2) or MgF(2), with the AlF(5)(2)(-) ions in the melt was considered. This association could be experimentally observed through a band broadening and a slight shift in the AlF(5)(2)(-) band frequency. Our vapor pressures and Raman data both indicate that MgF(2) clearly acts as an acid when added to NaF-AlF(3) melts of any composition. When CaF(2) is added, a slight decrease of vapor pressure occurs. Raman data indicate a decrease of AlF(4)(-) concentration, corresponding to a dissociation of CaF(2) with liberation of F(-) ions. All these results are, however, very much dependent on the initial melt composition. These data are explained in terms of acid-base, dilution, and association reactions of the solute with the solvent.

Raman spectroscopy and laser desorption mass spectrometry for minimal destructive forensic analysis of black and color inkjet printed documents.

Inkjet ink analysis is the best way to discriminate between printed documents, or even though more difficult, to connect an inkjet printed document with a brand or model of printers. Raman spectroscopy and laser desorption mass spectrometry (LDMS) have been demonstrated as powerful tools for dyes and pigments analysis, which are ink components. The aim of this work is to evaluate the aforementioned techniques for inkjet inks analysis in terms of discriminating power, information quality, and nondestructive capability. So, we investigated 10 different inkjet ink cartridges (primary colors and black), 7 from the HP manufacturer and one each from Epson, Canon and Lexmark. This paper demonstrates the capabilities of three methods: Raman spectroscopy, LDMS and MALDI-MS. Raman spectroscopy, as it is preferable to try the nondestructive approach first, is successfully adapted to the analysis of color printed documents in most cases. For analysis of color inkjet inks by LDMS, we show that a MALDI matrix (9-aminoacridine, 9AA) is needed to desorb and to ionize dyes from most inkjet inks (except Epson inks). Therefore, a method was developed to apply the 9AA MALDI matrix directly onto the piece of paper while avoiding analyte spreading. The obtained mass spectra are very discriminating and lead to information about ink additives and paper compositions. Discrimination of black inkjet printed documents is more difficult because of the common use of carbon black as the principal pigment. We show for the first time the possibility to discriminate between two black-printed documents coming from different, as well as from the same, manufacturers. Mass spectra recorded from black inks in positive ion mode LDMS detect polyethylene glycol polymers which have characteristic mass distributions and end groups. Moreover, software has been developed for rapid and objective comparison of the low mass range of these positive mode LDMS spectra which have characteristic unknown peaks.

Straightforward synthesis of conductive graphene/polymer nanocomposites from graphite oxide.

The reduction of graphite oxide (GO) in the presence of reactive poly(methyl methacrylate) (PMMA), under mild biphasic conditions, directly affords graphene grafted with PMMA. The resulting nanocomposite shows excellent electrical conductivities resulting from the optimal dispersion and exfoliation of graphene in the polymer matrix.

Stainless steel grafting of hyperbranched polymer brushes with an antibacterial activity: synthesis, characterization, and properties.

Two strategies were used for the preparation of hyperbranched polymer brushes with a high density of functional groups: (a) the cathodic electrografting of stainless steel by poly[2-(2-chloropropionate)ethyl acrylate] [poly(cPEA)], which was used as a macroinitiator for the atom transfer radical polymerization of an inimer, 2-(2-bromopropionate)ethyl acrylate in the presence or absence of heptadecafluorodecyl acrylate, (b) the grafting of preformed hyperbranched poly(ethyleneimine) onto poly(N-succinimidyl acrylate) previously electrografted onto stainless steel. The hyperbranched polymer, which contained either bromides or amines, was quaternized because the accordingly formed quaternary ammonium or pyridinium groups are known for antibacterial properties. The structure, chemical composition, and morphology of the quaternized and nonquaternized hyperbranched polymer brushes were characterized by ATR-FTIR reflectance, Raman spectroscopy, X-ray photoelectron spectroscopy, and atomic force microscopy. The peeling test confirmed that the grafted hyperbranched polymer films adhered much more strongly to stainless steel than the nongrafted solvent-cast films. The quaternized hyperbranched polymer brushes were more effective in preventing both protein adsorption and bacterial adhesion than quaternary ammonium containing poly(cPEA) primary films, more likely because of the higher hydrophilicity and density of cationic groups.

Gold-loaded carbon nanoparticles from poly(vinyl alcohol)-b-poly(acrylonitrile) non-shell-cross-linked micelles.

Herein we show that a new amphiphilic poly(vinyl alcohol)-b-poly(acrylonitrile) block copolymer dispersed in water can be easily loaded with gold nanoparticles by addition of chlorauric acid followed by reduction by sodium borohydride. After deposition of the so-loaded micelles onto a silicon wafer, followed by an appropriate thermal treatment, the poly(acrylonitrile) core of the micelles is carbonized, while the poly(vinyl alcohol) shell is completely decomposed and volatilized, leading to gold encapsulated in carbon nanoparticles. The morphology of the micelles is maintained during thermal treatment without requiring shell-cross-linking of the micelles prior to pyrolysis.

Association study between candidate genes and obesity-related phenotypes using a sample of lumberjacks.

INTRODUCTION: Complex traits such as obesity are modulated by genetic and environmental factors and lead to varied clinical presentations.The aim of this study was to investigate associations between candidate genes and obesity-related phenotypes using a sample of 252 lumberjacks issued from a founder population and sharing a common and circumscribed environment. METHODS: Thirty-seven variants in 18 genes were genotyped. The restriction fragment length polymorphism method and the template-directed dye-terminator incorporation assay with fluorescence polarization detection were employed for the genotyping assays. Multivariate logistic regression models were built in order to calculate the relative odds of exhibiting obesity-related phenotypes associated with the presence of the studied polymorphism. Among them, 21 single nucleotide polymorphisms were tested for associations with obesity phenotypes. RESULTS: Significant associations were found between carriers of the minor alleles of APOE-epsilon2, FABP2-A54T, UCP1-L229M, LPL-HindIII, LPL-S447X and LPL-T1973C, patients bearing a combination of LPL-D9N, LPL-N291S and LPL-P207L and obesity-related phenotypes. CONCLUSION: The present results suggest that a particular population such as lumberjacks, sharing the same environment, could help target genes involved in complex traits.

Combination of electrografting and atom-transfer radical polymerization for making the stainless steel surface antibacterial and protein antiadhesive.

A two-step "grafting from" method has been successfully carried out, which is based on the electrografting of polyacrylate chains containing an initiator for the atom transfer radical polymerization (ATRP) of 2-(tert-butylamino)ethyl methacrylate (TBAEMA) or copolymerization of TBAEMA with either monomethyl ether of poly(ethylene oxide) methacrylate (PEOMA) or acrylic acid (AA) or styrene. The chemisorption of this type of polymer brushes onto stainless steel surfaces has potential in orthopaedic surgery. These films have been characterized by ATR-FTIR, Raman spectroscopy, atomic force microscopy (AFM), and measurement of contact angles of water. The polymer formed in solution by ATRP and that one detached on purpose from the surface have been analyzed by size exclusion chromathography (SEC) and (1)H NMR spectroscopy. The strong adherence of the films onto stainless steel has been assessed by peeling tests. AFM analysis has shown that addition of hydrophilic comonomers to the grafted chains decreases the surface roughness. According to dynamic quartz crystal microbalance experiments, proteins (e.g., fibrinogen) are more effectively repelled whenever copolymer brushes contain neutral hydrophilic (PEOMA) co-units rather than negatively charged groups (PAA salt). Moreover, a 2- to 3-fold decrease in the fibrinogen adsorption is observed when TBAEMA is copolymerized with either PEOMA or AA rather than homopolymerized or copolymerized with styrene. Compared to the bare stainless steel surface, brushes of polyTBAEMA, poly(TBAEMA-co-PEOMA) and poly(TBAEMA-co-AA) decrease the bacteria adhesion by 3 to 4 orders of magnitude as revealed by Gram-positive bacteria S. aureus adhesion tests.

Determination of an acidic scale in room temperature ionic liquids.

The acidity scale of different Brønsted acids in ionic liquids such as [BMIM][NTf2], [BMIM][BF4], and [BMMIM][BF4] has been investigated by determination of Hammett functions, using a spectrophotometric indicator method. This scale should permit one to correlate the acidity strength of ionic liquid systems with their ability to achieve acid-catalyzed reactions.

Synthesis of copolymer brushes endowed with adhesion to stainless steel surfaces and antibacterial properties by controlled nitroxide-mediated radical polymerization.

Novel copolymer brushes have been synthesized by a two-step "grafting from" method that consists of the electrografting of poly(2-phenyl-2-(2,2,6,6-tetramethyl-piperidin-1-yloxy)-ethylacrylate) onto stainless steel, followed by the nitroxide-mediated radical polymerization of 2-(dimethylamino ethyl)acrylate and styrene or n-butyl acrylate, initiated from the electrografted polyacrylate chains. The grafted copolymers were quaternized in order to endow them with antibacterial properties. Peeling tests have confirmed the strong adhesion of the grafted copolymer onto the stainless steel substrate. Quartz crystal microbalance experiments have proven that fibrinogen adhesion is lower on the hydrophilic quaternized films compared to the nonionic counterpart. Such quaternized copolymers exhibit significant antibacterial activity against the Gram-positive bacteria S. aureus and the Gram-negative bacteria E. coli.