Background: Enterotoxigenic Escherichia coli (ETEC) strains producing multiple enterotoxins are important causes of post-weaning diarrhea (PWD) in pigs. The aim of the present study was to investigate the fecal presence of ETEC enterotoxin as well as F4 and F18 genes as an indicator of colistinsulfate (CS) efficacy for treatment of PWD in pigs. Forty-eight piglets were weaned at the age of 21 d, and were divided into four groups: challenged treated, challenged untreated, unchallenged treated, and unchallenged untreated. Challenge was performed using 10 9 CFU of an ETEC: F4 strain, and treatment was conducted using oral CS at the dose of 50,000 IU/kg. The fecal presence of genes encoding for STa, STb, LT, F4 and F18 was detected using PCR.
TP from pre-inoculation sample collection to post-inoculation sample collection served as an indication of dehydration.
After CS oral administration (30 d), blood samples (3 mL) were collected from the cannula at 30 min and 1, 2, 4, 6, 8, 12, 24, 36, and 48 h in potassium EDTA tubes. Plasma was separated by centrifugation at 2000 g for 10 min and stored at -20°C prior to analysis. Colistinsulfate plasma concentrations were determined by a liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). The HPLC system was a series 200 liquid chromatography apparatus (Perkin–Elmer, Boston, MA, USA) and the spectrometry system used was an API 2000 QTRAP AB-SCIEX (Concord, ON, Canada). Colistinsulfate was extracted from the pig plasma using a protein precipitation method; 200 μL of plasma was mixed with 200 μL of internal standard solution (500 ng/mL tylosine in acetonitrile) in a 1.5 mL centrifuge tube. Samples were vortexed and allowed to rest 10 min at room temperature prior to centrifugation. Samples were centrifuged at approximately 12,000 × g for 5 min and 200 µL of supernatant was transferred into an injection vial.
Because colistin is a cationic molecule, we looked for a medium which was deprived of any ions in its original composition and which was also deprived of any electro- lytes to avoid Proteus spp. swarming. The optimal condi- tions for LBJMR medium (growth of colistin- and vancomycin-resistant strains and inhibition of colistin- or vancomycin susceptible strains) were obtained on Purple Agar Base with glucose (Additional file 1: Table S3), 4 μg/mL of colistinsulfate and 50 μg/ml of vanco- mycin (Additional file 1: Table S4). The culture of ten chicken stools from Algeria enabled the detection of E. coli strains harboring the mcr-1 gene from three of the five samples which were positive for mcr-1 by RT-PCR, including strain 235 which had previously been isolated on Cepacia medium  and no Gram-negative strains were detected from the five negative samples (Additional file 1: Table S2). A correlation was observed between the Ct-values that reflect the DNA concentration and the re- sults of culture (Additional file 1: Table S2).
There is currently no commercial medium to screen gram-negative bacteria harboring the mcr-1 gene. Nor- dmann et al. (6) described an in-house SuperPolymyxin medium composed of eosin methylene blue agar, 3.5 mg/L of colistinsulfate, 10 mg/L of daptomycin, and 5 mg/L am- photericin B, which showed excellent sensitivity and speci- ficity. Colistin resistance can be confirmed within 2 h by us- ing an in-house rapid polymyxin Nordmann-Poirel test (7). The mcr-1 gene can be rapidly detected by real-time PCR of DNA extracts obtained from bacterial strains or directly from stool samples (2,8,9).
Here, we studied the impact of non-absorbed oral colistin on the emergence of CR in the gut microbiota of patients from the RGNOSIS-WP3 randomized controlled trial. 6 Thirty-nine subjects colonized with MDR-E were randomized to receive oral colistinsulfate 2 MIU 4 times a day + neomycin sulfate 500 mg bid for 5 days followed by a fecal microbiota transplant (FMT) from healthy donors, or no intervention. Stool samples were collected on visit V0 (screening sample), V2 (after 5 days of oral decontamination and before FMT for the intervention group), V3, V4 and V5, re- spectively 5–10 days, 30–40 days and 150–210 days later. 6 Stool samples from donors and 15 subjects from the intervention group and 15 from the control group were available for this work and plated on Drigalski plates (control) and SuperPolymyxin R plates. 5 Colony forming units (CFU) counts of all Gram-negative rods were determined. Isolates growing on Superpolymyxin R plates were identiﬁed by MALDI-TOF; CR was conﬁrmed by the culture-based Rapid Polymyxin NP test and MIC determined by the microdilu- tion method. The limit of detection was 10 2 CFU/g of stool. CR- E. coli were sequenced using the Illumina HiSeq technology. To deter- mine whether CR isolates were present before the intervention, a speciﬁc mcr-1 PCR was performed on patients stool prior to inter- vention (V0) and on the donor’s stool. 7 Electroporation of plasmids was performed to localize the gene conferring resistance to colistin and neomycin and molecular typing of the electroporants was per- formed using PCR based replicon typing (PBRT).
(NEM_7118432), Candida tropicalis (NEM_7114236), Candida krusei (ATCC6258; LGC Standards, Molsheim, France), Yarrowia lipolytica (NEM_7115511) and laboratory isolates of
S. cerevisiae (BY4741, BY4743 23 ) were used in this study. Fluconazole- and caspofungin-
resistant strains used in this study are listed in Table 1 and were obtained from D. Perlin (PHRI, New Jersey, USA) and D. Sanglard (CHUV, Lausanne, Switzerland). Strains were routinely grown at 30 °C in Yeast Peptone Dextrose medium (YPD; 1% yeast extract, 2% peptone, 2% glucose), or Synthetic Dextrose minimal medium (SD; 0.67% yeast nitrogen base without amino acids (Difco), 2% glucose) and 2% agar for solid media. RPMI 1640 glutamax medium (Invitrogen) buffered with 50 mM Hepes (4-(2-hydroxyethyl)-1- piperazineethanesulfonic acid), pH 7 was used for screening compounds that synergize with aminocandin and for chequerboard assays. Alternatively RPMI 1640 glutamax medium buffered with 0.165 M MOPS (3-(N-morpholino)propanesulfonic acid) was used. Aminocandin (a kind gift from Dr. Dominique Lebeller), caspofungin (MSD), colistinsulfate (Sigma-Aldrich, Saint-Quentin Fallavier, France), colistin sodium methanesulfonate (Sigma- Aldrich), hygromycin B (Sigma-Aldrich), nourseothricin (Werner BioAgents, Jena, Germany) and 5-fluorocytosine (Sigma-Aldrich) were dissolved in H 2 O. Fluconazole (Sigma-Aldrich)
Acinetobacter baumannii, Pseudomonas aeruginosa, and the carbapenemase-producing K. pneumoniae was successfully treated by a 20-day course of colistin. The patient was
then repatriated to an intensive care unit in France, where two E. coli isolates desig- nated WI1 and WI2 were recovered from a feces CPE screening sample taken on the patient’s admission with CarbaSmart medium (bioMérieux, La Balme, France).
Cement is a strategic commodity in the civil engineering for the construction of reinforced concrete structures. But its production generates around 5% of toxic gases such as CO 2 responsible for environmental degradation. Furthermore, cement industry is a consumer sector of non-renewable energy. The use in the cement of natural additions is a solution to reduce the CO 2 gas and the cost of production. The purpose of this work is the study of a sustainable building material: natural pozzolan Beni-saf (PNB) incorporated to mortars exposed to sulfate attack (5% Na 2 SO 4 ). The loss of mass, monitoring the pH reading of each attack solution as well as specimens dimensions are different tests to study the durability of mortars made with 10, 20 and 30% of natural pozzolan. The result derived from this research is that pozzolan improves mortars resistance to sodium sulfate environment.
In our previous study, 3 a series of thermosensitive hydrogels have been developed for the buccal delivery of salbutamol sulfate (SS). As a short-acting beta-2 adrenoceptor agonist, SS is widely used in the treatment of reactive airways diseases such as asthmatic disorders and chronic obstructive pulmonary diseases (COPD). The hydrogels are based on a mixture of poloxamer analogs and xanthan gum, which present a synergy of thermosensitive and mucoadhesive properties. Poloxamers, are triblock copolymers of PEO-PPO-PEO (polyethyleneoxy-polypropyleneoxy-polyethyleneoxy), which self-assemble as micelles in solution. 4 The gelation is a result of inter-micellar entanglement and packing when temperature increases. 5 In our previous study, two water soluble poloxamers of different PEO/PPO ratios: P407, P188 were used to prepare the thermosensitive base. Satiaxane (xanthan gum) was added to obtain suitable mechanical and mucoadhesive properties. The previous results showed that the combination of P407, P188 and xanthan gum provided not only an optimal sol-gel transition temperature, but also an increased gel strength, which sustained drug release, thus providing potential useful clinic interests.
Aix-Marseille Université, Université Sud Toulon, CNRS/INSU, IRD, MIO-UM110, Marseille, France a ; Aix-Marseille Université, CNRS, LCB-UMR7283, Marseille, France b ;
Laboratoire de Finition CEA, Institut de Génomique-Génoscope, Évry, France c ; Laboratoire d’Analyses Bioinformatiques pour la Génomique et le Métabolisme (LABGeM),
Genoscope-IG-CEA, Évry, France d ; CNRS, UMR7156, Université Louis Pasteur, Strasbourg, France e
Here, we report the draft genome sequence of Desulfotomaculum hydrothermale, a sulfate-reducing, spore-forming bacterium isolated from a Tunisian hot spring. The genome is composed of 2.7 Mb, with a GⴙC content of 49.48%, and it contains 2,643 protein-coding sequences.
aerosols, changing physicochemical properties, and decreasing multiphase reactivity caused by IEPOX multiphase chemistry have yet to be updated in the regional- and global-scale models to properly predict IEPOX-SOA formation and its impact on air quality and climate. In isoprene-dominated areas, IEPOX-OS, and potentially other biogenic/anthropogenic OS, likely govern the physicochemical properties of aerosol as well as the distribution of inorganic species such as sulfate or ammonium. Consequently, aerosol growth, multiphase reactions, including aging and reactive uptake of other species, and CCN activity change as surface tension, acidity, hygroscopicity and viscosity are modified. These changes could greatly impact atmospheric composition of biogenic SOA formed over isoprene-dominated areas. Hence, changes in SO 2 emissions at different locations around the world over time
and lasted for a prolonged interval (>1 Gyr) after GOE I (Bekker and Holland, 2012; Lyons et al., 2014; Planavsky et al., 2014) .
Oceanic sulfate plays a key role in the biogeochemical cycles of S, C, O, and Fe (Planavsky et al., 2012; Shen et al., 2002) , e.g., via the role of microbial sulfate reduction (MSR) in remineralization of organic matter and oxidation of the sedimentary methane flux (Jørgensen, 1982; Valentine, 2002) . Oceanic sulfate concentrations are also an important proxy for seawater chemistry and the oxygenation history of the Precambrian atmosphere and oceans (Canfield, 2004; Kah et al., 2004; Kah and Bartley, 2011; Scott et al., 2014) . The Archean ocean is thought to have had very low sulfate concentrations due to extremely low pO 2 , possibly <0.2 mM, as inferred from
optimal conditions, thus interact electrostatically with anionic HSPGs and can enter into cells via endocytosis [301–305,308]. Interestingly, it was showed that this interaction is not solely governed by electrostatic interactions. Indeed, the affinity between sulfated GAGs and polyplexes is possibly dependant on the nature of the polymer, suggesting that other interactions may occur . If the absence of sulfate inhibits gene transfer efficiency , a high density of HS promotes polyplex uptake in mammalian cells [304,307]. However, the binding of polyplexes are not always correlated to their internalization [302,308] nor to their intracellular path . This suggests that uptake and transgene expression are function of the relative abundance of different GAG-carrying proteins rather than the overall HSPG level . HSPGs comprise an assorted group of proteins that contain covalently linked heparan sulfate (HS) glycosaminoglycan (GAG) chains. There are two major families of HSPGs that are largely expressed at the surface of mammalian cells, namely the syndecans composed of four members (SDC1-4) and the glypicans that consist of six members (GPC1-6). Whereas the core protein of SDCs is type I integral membrane protein exhibiting a short cytoplasmic tail, the core protein of GPCs possesses a glycosyl phosphatidylinositol (GPI) anchored extracellular domain attached at their carboxyl terminus . In the context of gene delivery, the role of HSPGs at the cell surface is mainly studied as a whole. The striking difference in structure between SDCs and GPCs may suggest that they are not involved to the same extent in the binding and internalization of polyplexes. While some studies have demonstrated that SDCs can be understood as receptors for polyplexes capable of driving their endocytosis [303,305,308], the role of GPCs in gene transfer process has not been investigated yet.
Mann-Whitney test. P values <0.05 were considered statistically different using GraphPad Prism 7.
RESULTS AND DISCUSSION
As shown in Figure 1A and Supplementary Figure S1, the mass spectrum of colistin susceptible S. enterica is dominated by a set of three of peaks assigned to bis-phosphorylated hexa- acyl lipid A, tri-phosphorylated hexa-acyl lipid A and bis- phosphorylated hepta-acyl lipid A. The major peaks at m/z 1796.2 and m/z 1876.2 correspond to hexa-acyl diphosphoryl and hexa-acyl triphosphoryl lipid A, respectively, containing four C14:0 3-OH, one C14:0 and one C12:0. The peak at m/z 2034.2 corresponds to hepta-acyl diphosphoryl lipid A four C14:0 3-OH, one C14:0, one C12:0 and one C16:0. Peaks at m/z 1812.2 and m/z 2050.2 are also observed in colistin susceptible Salmonella enterica and which can be tentatively be assigned to hexa-acyl diphosphoryl lipid A, containing five C14:0 3-OH and one C12:0, and hepta-acyl diphosphoryl lipid A five C14:0 3-OH, one C12:0 and one C16:0, respectively.
Ultrasound-assisted Conversion of ThO 2 into a Th peroxo sulfate L. Bonato, 1 M. Virot, 1 T. Dumas, 2 P. Moisy, 2 S. I. Nikitenko 1
1 Institut de Chimie Séparative de Marcoule (ICSM) – UMR5257, CNRS/CEA/UM/ENSCM, Site de Marcoule, 30207 Ba-
gnols sur Cèze, France
première partie de la réaction (jusqu’à α = 0,5) et un modèle R 1 pour la seconde. La constante
apparente de vitesse k app diminue quand la masse de l’échantillon augmente.
Modestov et al. [Mod01] ont étudié la déshydratation du sulfate de lithium monohydraté sur des monocristaux (~ 1 cm) dans la direction . Considérant que le front de réaction (c’est-à-dire la zone dans laquelle se déroule la réaction) a une épaisseur d’environ 100 µm, d’après les résultats de [Kir88], les auteurs proposent d’effectuer la réaction sur un échantillon plus gros afin que le front de réaction atteigne 100µm et qu’il puisse progresser de manière stationnaire. Ces études sont réalisées en conditions non-isothermes, sous vide. Les résultats sont présentés sous forme de courbe représentant la profondeur du front de réaction en fonction de la température. Ils montrent une dépendance quadratique de cette profondeur par rapport à la température. Des expériences menées en conditions isothermes fournissent une valeur de l’énergie d’activation environ deux fois plus faible que celles obtenues par les autres auteurs.
The effect of CS on MSC in biomaterials have also been explored. MSC proliferation has been studied on GAG-derivatized chitosan membranes and it was found that MSC growth increased as much as fivefold on GAG-immobilized membranes in comparison to normal tissue culture plastic or only chitosan. Results exhibit the highest cell density when membranes were prepared with chondroitin sulfate vs other GAGs (Uygun, Stojsih et Matthew, 2009). PEG/CS hydrogels created by photopolymerization of pre-functionalized CS with methacrylate groups have proven to provide a microenvironment that is conducive for MSC chondrogenesis, facilitating condensation of encapsulated MSC followed by early expression of cartilage specific markers and matrix component production (Varghese et al., 2008). PEG/CS hydrogels with incorporated bio-functional building blocks such as RGD peptides allow tridimensional culture and expansion on MSC. Additionally, the CS based hydrogel exhibited a binding to bone morphogenetic protein-2 (BMP-2), which mediated MSC osteogenic differentiation, indicating its potential in bone tissue regeneration (Anjum et al., 2006). Silk fibroin/gelatin–chondroitin sulfate–hyaluronic acid scaffolds have been previously fabricated providing a supportive structure and mimetic cartilage environment for MSC chondrogenesis, enabling cartilage regeneration (Sawatjui et al., 2015). The previous studies show the potential of CS in biomaterials with MSC for tissue engineering, as for example in regeneration of bone or cartilage.