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Atmospheric simulation chambers, are unique tools for investigating atmospheric processes in the gas and heterogeneous phases. They can provide a controlled yet realistic environment that simulates atmospheric conditions. In the current study, a Teflon atmospheric simulation chamber of 600 L, named THALAMOS (thermally regulated atmospheric simulation chamber) has been developed and cross-validated. THALAMOS can be operated over the temperature range 233 to 373 K under both static and flow conditions. It is equipped with state of the art instrumentation (selective ion flow tube mass spectrometry (SIFT-MS), long path Fourier transform infrared spectroscopy (FTIR), gas chromatography-mass spectrometry (GC-MS), various analyzers) for the in-line monitoring of both reactants and products. THALAMOS was validated by measuring the rate coefficients of well documented reactions, i.e. the reaction of ethanol with OH radicals and the reaction of dichloromethane with Cl atoms, in a wide temperature range. Two different detection techniques were used in parallel, FTIR and SIFT-MS, to internally cross-validate the obtained results. The measured rate coefficients are in excellent agreement, both between each other and with the literature recommended values. Furthermore, the gas phase oxidation of toluene by Cl atoms (kinetics and product yields) was studied in the temperature range of 253 to 333 K. To the best of our knowledge, THALAMOS is a unique facility on national level and among a few smog chambers internationally that can be operated in such a wide temperature range providing the scientific community with a versatile tool to simulate both outdoor and indoor physicochemical processes.

Hepatocyte Nuclear Factor-1A (HNF1A) is a master regulator of key glucose-responsive genes in metabolic organs such as the intestine, liver, kidney, and pancreas. Heterozygous mutations in HNF1A cause Maturity-Onset-Diabetes-of-the-Young (HNF1A-MODY) . Mutant carriers are normoglycemic in childhood, but with age, they develop hyperglycemia. Although insulin secretory defects have been extensively studied, it is unclear whether HNF1A deficiency contributes to a progressive decline in alpha cell function. HNF1A-MODY patients display glycosuria and endogenous glucose production linked to reduced renal SGLT2 expression. Since SGLT2 is also expressed in alpha cells and is required for glucagon regulation, we hypothesized that HNF1A coordinates renal and alpha cell SGLT2 activity to maintain glucose homeostasis. Here, we studied Hnf1a regulation of renal Sglt2, Sglt1, and Glut2 in Hnf1a-/- mutant mice, glucagon expression and secretion in siHNF1A transfected human islets and we developed a novel Hnf1a mutant mouse model (Hnf1a+/Δe4-10) . Sglt2 protein was significantly reduced in the kidney of Hnf1a-/- mice, with no changes in Sglt1 or Glut2 protein expression. We observed that HNF1A is heterogeneously expressed in alpha, beta, and delta cells. siHNF1A islets displayed reduced insulin secretion and content, which was rescued by GLP1 treatment. HNF1A deficiency reduced SGLT2 levels, correlating with increased glucagon content and secretion. The Hnf1a+/Δe4-mice were born normoglycemic but became glucose intolerant at 12 weeks of age, mimicking the clinical characteristics of HNF1A-MODY patients. Collectively, these findings indicate that HNF1A deficiency downregulates renal and alpha cell SGLT2 expression, which is associated with an alteration in glucagon secretion in response to glucose stimulation. The new Hnf1a+/Δe4-mouse model may be a useful tool to study the progression of MODY with and without chronic drug treatment. Disclosure A.Acosta-montalvo: None. V.Gmyr: None. M.Cnop: None. J.A.Kerr-conte: None. F.Pattou: None. M.Pontoglio: None. A.Liston: None. C.Bonner: None. C.Saponaro: None. J.Thevenet: None. M.Chiral: None. A.Piron: None. N.Delalleau: None. G.Pasquetti: None. A.Coddeville: None. M.Moreno: None. Funding EFSD/Lilly (2017) awarded to Caroline Bonner, I-SITE ULNE awarded to Caroline Bonner, the European Genomic Institute for Diabetes (ANR-10-LABX-46) , the European Consortium for Islet Transplantation funded by the Juvenile Diabetes Research Foundation. TIGER was developed within the European Union’s Horizon 2020 research and innovation program project T2DSystems, under grant agreement 667191 (to Miriam Cnop) .

Welding fumes (WF) are a complex mixture of ultrafine polymetallic particles, which can deposit in all regions of the respiratory tract. They contain high levels of transition metals (Cr, Cu, Mn …) known to catalyze the production of reactive oxygen species which are related to numerous adverse health effects. In industrial hygiene, collected WF are analyzed several days later, following usual standard techniques. However, during storage WF samples may undergo several physico-chemical modifications that may lead to an underestimation of their oxidative potential (OP) and metal bioaccessibility. In this work, we determined the influence of several usual storage conditions on the OP (dithiothreitol method OPDTT) and metal bioaccessibility of WF. Fresh WF were generated using a controlled welding bench and the collected particles were subsequently stored following various conservation conditions (temperature, atmosphere composition and durations) before analysis. OPDTT of WF significantly decrease with increasing storage duration when stored in ambient air, either at +20°C or -20°C, while there are no significant variations for storage under N2(g). Furthermore, the metal bioaccessibilities show similar trends to that of the OPDTT of WF for the tested conservation conditions. This work confirms that storage conditions of WF have an undeniable influence on their metal bioaccessibility and OPDTT. On-line analysis could offer an alternative to WF filter sampling, using OPDTT as a probe of health monitoring for exposed professionals

Indoor environmental exposure to pesticides has become one of the major concerns that might adversely affect human health and development. People spend most of their lifetime in enclosed indoor environments where they might inhale harmful toxic chemicals, such as pesticides, dispersed either in particulate or in a gas phase. In this study, an assessment of pesticide contamination in indoor environments was conducted. The study covered nine houses during one year, starting from February 2016 and ending in February 2017, in which both air and dust samples were assessed for their potential contamination with 50 pesticides. The results showed that all the assessed houses were contaminated by several pesticides, especially with the allethrin pesticide (detection frequency (DF) = 100%). The highest pesticide contamination was detected in the spring/summer season when it reached an average of around 185 ng g−1 and 186.4 ng sampler−1 in the collected dust and air samples, respectively. The potential contamination of pyrethroid insecticides within all the targeted samples revealed by this study stresses the importance of minimizing the use of such indoor treatments as part of the efficient prevention and control of human exposure to pesticides.

In order to clarify whether the mitochondrial dysfunction is closely related to the cell homeostasis maintenance after particulate matter (PM2.5) exposure, oxidative, inflammatory, apoptotic and mitochondrial endpoints were carefully studied in human bronchial epithelial BEAS-2B, normal human bronchial epithelial (NHBE) and chronic obstructive pulmonary disease (COPD)-diseased human bronchial epithelial (DHBE) cells acutely or repeatedly exposed to air pollution-derived PM2.5. Some modifications of the mitochondrial morphology were observed within all these cell models repeatedly exposed to the highest dose of PM2.5. Dose- and exposure-dependent oxidative damages were reported in BEAS-2B, NHBE and particularly COPD-DHBE cells acutely or repeatedly exposed to PM2.5. Nuclear factor erythroid 2-p45 related factor 2 (NRF2) gene expression and binding activity, together with the mRNA levels of some NRF2 target genes, were directly related to the number of exposures for the lowest PM2.5 dose (i.e., 2 μg/cm2), but, surprisingly, inversely related to the number of exposures for the highest dose (i.e., 10 μg/cm2). There were dose- and exposure-dependent increases of both nuclear factor kappa-B (NF-κB) binding activity and NF-κB target cytokine secretion in BEAS-2B, NHBE and particularly COPD-DHBE cells exposed to PM2.5. Mitochondrial ROS production, membrane potential depolarization, oxidative phosphorylation, and ATP production were significantly altered in all the cell models repeatedly exposed to the highest dose of PM2.5. Collectively, our results indicate a cytosolic ROS overproduction, inducing oxidative damage and activating oxygen sensitive NRF2 and NF-kB signaling pathways for all the cell models acutely or repeatedly exposed to PM2.5. However, one of the important highlight of our findings is that the prolonged and repeated exposure in BEAS-2B, NHBE and in particular sensible COPD-DHBE cells further caused an oxidative boost able to partially inactivate the NRF2 signaling pathway and to critically impair mitochondrial redox homeostasis, thereby producing a persistent mitochondrial dysfunction and a lowering cell energy supply.

A direct method to study essential genes is to construct conditional knock-down mutants by replacement of their native promoter by an inducible one. In Mycobacteria, replacement of an essential gene promoter with an anhydrotetracycline inducible one was successfully used but required a multi-step approach. In this work, we describe a gene cassette for the engineering of a conditional knock-down mutant, which allows the one-step targeted replacement of mycobacterial promoters by an anhydrotetracycline-inducible promoter. The functionality of this cassette was successfully tested by engineering conditional clpP and SecA1 mutants of Mycobacterium smegmatis.

A method coupling automated thermal desorption (ATD) with in-tube derivatization and solid phase micro-extraction (SPME), with on-injector derivatization gas chromatography - tandem mass spectrometry (GC/MSMS) was developed and used for the simultaneous quantification of 100 semivolatile (SVOCs) pollutants (50 pesticides, 16 PAHs, 22 PCBs, 7 phthalates and 5 alkylphenols) in indoor air, indoor dust, and for bioaccessibility evaluation. Chemical compounds in indoor air were sampled on Tenax-TA passive samplers (PAS) exposed for 15 days, while dust was collected on silicon carbide (SiC©) foam using a custom made vacuum cleaner head. After sampling, Tenax tubes from PAS were introduced directly into a thermal desorption tube while dust was extracted using accelerated solvent extractor (ASE). Extract is then used to spike clean Tenax-TA tube that is then treated as a Tenax from PAS. The derivatization agent (MtBSTFA) and a mix of internal standards were added into the tubes before thermal desorption and analysis by GC/MSMS. The SPME technique was applied to extract the 100 SVOCs from the different tested bioaccessibility solutions (saliva, gastric, intestinal). This method permits the determination of 100 pollutants in a single run, including those requiring a derivatization step, with very low quantification and detection limits. Indeed, ATD-GC/MSMS technic presents very low LOD (0.002 to 0.17 ng) and LOQ (0.007 to 0.56 ng) values while SPME-GC/MSMS displays slightly higher LOD (0.044 to 12.3 ng) and LOQ (0.10 to 37.0 ng) ranges.

This study provides the first kinetic and mechanistic study of the photolysis of 4-hydroxy-4-methyl-2-pentanone (4H4M2P) and the determination of the temperature dependence of the relative rate coefficient for the reaction of OH radicals with 4H4M2P. The UV absorption spectrum of 4H4M2P was determined in the spectral range 200–360 nm. The photolysis frequency of this compound in the atmosphere was evaluated relative to actinometers and found to be J 4 H 4 M 2 P a t m = 4.2 × 10 − 3 h − 1 , corresponding to a lifetime of about 10 days. Using 4H4M2P cross section measurements, an atmospheric effective quantum yield of 0.15 was calculated. The main primary photolysis products were acetone (121 ± 4) % and formaldehyde (20 ± 1) %. A low methanol yield of (3.0 ± 0.3) % was also determined. These results enabled us to propose a mechanistic scheme for the photolysis. Rate coefficients for the reaction of 4H4M2P with OH radicals were determined over the temperature range 298–354 K and the following Arrhenius expression was obtained: kOH+4M4H2P = (1.12 ± 0.40) × 10−12exp(461.5 ± 60/T) cm3 molecule−1 s−1. The lifetimes of 4H4M2P due to reaction with OH radicals has been estimated to ∼2.5 days and indicates that the gas-phase reaction with the OH could be the main loss process for this compound.

Carbonyls play an important role in atmospheric chemistry due to their formation in the photooxidation of biogenic and anthropogenic volatile organic compounds and their high atmospheric reactivity. The Cl-initiated kinetics of two α-diketones (2,3-pentanedione (PTD) and 2,3-hexanedione (HEX)) have been determined as well as the OH + HEX rate constant using atmospheric simulation chamber experiments and the relative rate method. Up to three different reference compounds were used to assess robust results. The following rate constants (in cm3 molecule−1 s−1) have been obtained at 298 K: k(Cl + PTD) = (1.6 ± 0.2) × 10−11, k(Cl + HEX) = (8.8 ± 0.4) × 10−11, and k(OH + HEX) = (3.6 ± 0.7) × 10−12 with a global uncertainty of 30%. The present determinations of Cl- and OH- reaction rate constants for HEX constitute first measurements. Using the present measurements, a recently improved structure–activity relationship for Cl + ketone reactions has been updated by introducing an F(–COCO–) factor of 8.33 × 10−4. Atmospheric lifetime calculations indicate that chlorine-initiated oxidation may be a significant α-diketone sink in the marine-boundary layer or in places where high Cl concentrations may be found.

A method coupling automated thermal desorption and gas chromatography-tandem mass spectrometry was developed and used for the quantification of 16 PAHs (acenaphthene, anthracene, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[e]pyrene, benzo[g,h,i]perylene, benzo[k]fluoranthene, chrysene, dibenzo[a,h]anthracene, fluoranthene, fluorene, indeno(1, 2, 3)pyrene, naphthalene, phenanthrene and pyrene) in indoor air and dust. Polycyclic aromatic hydrocarbons (PAHs) in indoor air were sampled using Tenax-TA passive samplers (PAS) exposed during 14 days. Indoor dust, collected on filters using a modified vacuum cleaner head equipped with a sieving device, was first extracted using accelerated solvent extractor. The extract was then concentrated and deposed on a Tenax-TA tube prior to analysis. Using the same thermal desorption method for the analysis of two different matrices gives complementary results with good sensitivity. Limit of detection varied between 0.009 and 0.031 ng and between...

While the knowledge of the underlying mechanisms by which air pollution-derived particulate matter (PM) exerts its harmful health effects is still incomplete, detailed in vitro studies are highly needed. With the aim of getting closer to the human in vivo conditions and better integrating a number of factors related to pre-existing chronic pulmonary inflammatory, we sought to develop primary cultures of normal human bronchial epithelial (NHBE) cells and chronic obstructive pulmonary disease (COPD)-diseased human bronchial epithelial (DHBE) cells, grown at the air-liquid interface. Pan-cytokeratin and MUC5AC immunostaining confirmed the specific cell-types of both these healthy and diseased cell models and showed they are closed to human bronchial epithelia. Thereafter, healthy and diseased cells were repeatedly exposed to air pollution-derived PM4 at the non-cytotoxic concentration of 5 μg/cm2. The differences between the oxidative and inflammatory states in non-exposed NHBE and COPD-DHBE cells indicated that diseased cells conserved their specific physiopathological characteristics. Increases in both oxidative damage and cytokine secretion were reported in repeatedly exposed NHBE cells and particularly in COPD-DHBE cells. Diseased cells repeatedly exposed had lower capacities to metabolize the organic chemicals-coated onto the air-pollution-derived PM4, such as benzo[a]pyrene (B[a]P), but showed higher sensibility to the formation of OH-B[a]P DNA adducts, because their diseased state possibly affected their defenses. Differential profiles of epigenetic hallmarks (i.e., global DNA hypomethylation, P16 promoter hypermethylation, telomere length shortening, telomerase activation, and histone H3 modifications) occurred in repeatedly exposed NHBE and particularly in COPD-DHBE cells. Taken together, these results closely supported the highest responsiveness of COPD-DHBE cells to a repeated exposure to air pollution-derived PM4. The use of these innovative in vitro exposure systems such as NHBE and COPD-DHBE cells could therefore be consider as a very useful and powerful promising tool in the field of the respiratory toxicology, taking into account sensitive individuals.

Indoor air quality (IAQ) receives an increasing attention because of long-term occupancy of confined environments with high specific pollutant concentrations, especially volatile organic compounds, inducing a risk for human health. Standard air quality instruments are not adapted for long-term continuous real-time monitoring of these environments, and a promising alternative resides in using electronic gas sensors, which are miniature and low-cost devices capable to detect air pollutants, even at low concentrations. The present study focuses on IAQ monitoring with electronic gas multi sensor systems, as well as with typical indoor air analyzers in a 40 m 3 experimental room. The suitability of electronic gas sensors to monitor pollution events in indoor environments, characterized by transient concentrations of VOCs, has been characterized by simultaneous injections of acetaldehyde, acetone, formaldehyde, toluene and o -xylene. The impact of a green wall structure in the room has been assessed by the reduction of residual pollutant and higher decay of injected oxygenated VOCs than in the empty room. It has been shown that electronic gas sensor response show a good correlation within the analyzer measurements, both for injected VOC concentrations and their decay. Indeed, these systems provide relevant information for air treatment control system and detection of indoor air quality events, in the case of simple gas matrixes, however realistic complex matrixes are poorly monitored by electronic gas sensors selected in this study. This limitation could be overcome by increasing the number of sensitive sensors to indoor air specific composition.

Using a combination of the open circuit potential and pH profiles of aniline (An) polymerization and their mathematical treatment, we develop a new convenient semiquantitative approach to determine the influence of the dispersed nanoparticles (e.g., TiO2 nanoparticles) on the kinetic features of this process and molecular mass of the formed polyaniline (PANI). It is revealed that the reciprocal values of the polymerization stages, namely, the duration of the induction period, of homogeneous and heterogeneous pernigraniline (PN) accumulation, and of PN reduction with An, are linear functions of the weight fraction of the nanoparticles. We found that when nanoparticles are added the weight-averaged molecular weight of PANI initially increases from 56 000 to 79 000 and the polydispersity index drops from 3.9 to 1.7. However, at high TiO2 concentrations, the former dramatically decreases, whereas the latter increases. We use the relative proton concentration as a function of time and the different extents of ...

The significant effect of various aromatic sulfonic acids on peculiarities of chemical polymerization of aniline in the water dispersion of polycarbonate (PC) powder, morphology and properties of the formed PC-polyaniline composites has been found and studied. In particular, we demonstrate that the rate of the aniline polymerization and polyaniline yield strongly depend on the size and surface activity of the acid-dopant anions. It is found that in the case of the large dopants (e.g. dodecylbenzenesulfonic acid) the composites unexpectedly have the reduced thermostability while the composites with the small dopants (e.g. p -toluenesulfonic acid or 2-naphtalenesulfonic acid) display the enhanced thermal stability as compared with the pure PC. We confirm that the acid-dopant nature affects the thermal stability of conductivity of the compression molded composite films. The solution-cast composite films have the dopant dependent sensitivity to gaseous ammonia in a wide ppm range of its concentrations.

In order to clarify whether the mitochondrial dysfunction is closely related to the cell homeostasis maintenance after particulate matter (PM

Passive samplers have recently been proposed as simple and inexpensive tools to measure emissions of Volatile Organic Compounds (VOCs) from building and furnishing materials. These samplers can be used to pinpoint strong emitters of targeted pollutants, including hydrocarbons and oxygenated VOCs, which is of great interest to design efficient strategies aimed at improving indoor air quality. A passive sampler consists of a small cell that is exposed on a flat surface to trap material emissions. Three Passive Flux Samplers (PFS) have been developed at Mines Douai, an engineering school from Northern France, to carry out source apportionment studies of formaldehyde, acetaldehyde, and aromatic hydrocarbons, including benzene, toluene, xylenes, and higher molecular weight compounds. Over a 6-h exposure duration, these PFS exhibit linear responses and detection limits of a few μg m −2 h −1 that are low enough for monitoring material emissions and to perform extensive source apportionment studies. A few other samplers, designed using different geometries, have also been proposed in the literature. This publication summarizes findings on the development and the use of passive samplers with the objective to highlight the potential of these new tools for indoor air quality studies.

The gas-phase ozonolysis of three methylated alkenes, i.e., trans-2,2-dimethyl-3-hexene (22dM3H), trans-2,5-dimethyl-3-hexene (25dM3H), and 4-methyl-1-pentene (4M1P), has been investigated in the presence of sufficient hydroxyl radical scavenger in a laminar flow reactor at ambient temperature (296 ± 2 K) and P = 1 atm of dry air (RH ≤ 5%). Ozone levels in the reactor were monitored by an automatic analyzer. Alkene and gas-phase product concentrations were determined via online sampling either on three-bed adsorbent cartridges followed by thermodesorption and GC/FID-MS analysis or on 2,4-dinitrophenylhydrazine (DNPH) cartridges for subsequent HPLC/UV analysis. Reaction rate coefficients of (3.38 ± 0.12) × 10−17 for 22dM3H and (2.71 ± 0.26) × 10−17 for 25dM3H, both in cm3 molecule−1 s−1 units, have been obtained under pseudo–first-order conditions. Primary carbonyl products have been identified for the three investigated alkenes, and branching ratios are reported. In the case of 4M1P ozonolysis, the yield of a Criegee intermediate was indirectly determined. Kinetics and product study results are compared to those of literature when available. This work represents the first investigation of reaction products in the ozonolysis of 22dM3H, 25dM3H, and 4M1P in a flow reactor.

New ammonia and amine sensing materials based on hybrid polyaniline/titanium dioxide nanocomposites were synthesized by a one-pot chemical polymerization. Particular attention was paid to the influence of TiO2 nanoparticles and the surface active dopants on the structure–property relationship of the nanocomposites in terms of their morphology, composition, electrical and sensing properties. Dodecylbenzenesulfonic (DBSA) and lauryl sulfuric (LSA) acids were used as polyaniline dopants. The nanocomposites sensing properties were evaluated at different humidity levels when exposed to ammonia, methyl- and trimethylamine (0.2 ppm to 1 ppm). The materials demonstrated strong responses and high sensitivity to the gases with a quantification limit of 20 ppb for ammonia. The sensors are reversible and have short response times. However, the response magnitude depends on the dopant nature. The results were interpreted in terms of basicity and size of the analyte gas molecules.

SecB chaperones assist protein export in bacteria. However, certain SecB family members have diverged to become specialized toward the control of toxin-antitoxin (TA) systems known to promote bacterial adaptation to stress and persistence. In such tripartite TA-chaperone (TAC) systems, the chaperone was shown to assist folding and to prevent degradation of its cognate antitoxin, thus facilitating inhibition of the toxin. Here, we used both the export chaperone SecB of Escherichia coli and the tripartite TAC system of Mycobacterium tuberculosis as a model to investigate how generic chaperones can specialize toward the control of TA systems. Through directed evolution of SecB, we have identified and characterized mutations that specifically improve the ability of SecB to control our model TA system without affecting its function in protein export. Such a remarkable plasticity of SecB chaperone function suggests that its substrate binding surface can be readily remodeled to accommodate specific clients.

Hybrid polyaniline (PANI) nanocomposites were prepared by chemical oxidative polymerization of aniline in the presence of different metal oxide nanoparticles (TiO 2 in rutile and anatase forms, SnO 2 ). Influence of the near ultraviolet (UV) light on synthesized materials properties and possibility of polymer phase photocatalytic degradation were investigated. Specifically, we have studied the nanocomposites' properties before and after the UV exposition by UV-Vis spectroscopy, thermogravimetric analysis and conductivity measurements. The obtained results suggest that synthesized materials undergo solid-state photodegradation depending on the energy gap between PANI LUMO and metal oxides conduction band. A possible photodegradation mechanism based on the sensitizing effect by PANI is proposed.

International audience; The kinetics of the reactions of limonene with OH and OD radicals has been studied using a low-pressure flow tube reactor coupled with a quadrupole mass spectrometer: OH + C10H16 → products (1), OD + C10H16 → products (2). The rate constants of the title reactions were determined using four different approaches: either monitoring the kinetics of OH (OD) radicals or limonene consumption in excess of limonene or of the radicals, respectively (absolute method), and by the relative rate method using either the reaction OH (OD) + Br2 or OH (OD) + DMDS (dimethyl disulfide) as the reference one and following HOBr (DOBr) formation or DMDS and limonene consumption, respectively. As a result of the absolute and relative measurements, the overall rate coefficients, k1 = (3.0 ± 0.5) × 10–11 exp((515 ± 50)/T) and k2 = (2.5 ± 0.6) × 10–11 exp((575 ± 60)/T) cm3 molecule–1 s–1, were determined at a pressure of 1 Torr of helium over the temperature ranges 220–360 and 233–353 K, respectively. k1 was found to be pressure independent over the range 0.5–5 Torr. There are two possible pathways for the reaction between OH (OD) and limonene: addition of the radical to one of the limonene double bonds (reactions 1a and 2a) and abstraction of a hydrogen atom (reactions 1b and 2b), resulting in the formation of H2O (HOD). Measurements of the HOD yield as a function of temperature led to the following branching ratio of the H atom abstraction channel: k2b/k2 = (0.07 ± 0.03) × exp((460 ± 140)/T) for T = (253–355) K.

Commercial miniature gas sensors, because they are smaller and cheaper than conventional instruments, can be deployed in large numbers to investigate indoor air quality, for research and operational purposes. To compensate for their limited metrological performances, it is necessary to develop relevant data treatment procedures. We applied an unsupervised classification approach based on the bisecting K-means algorithm to data acquired by online gas analyzers and by miniature sensors during a measurement campaign in a low energy school building. This procedure, applied to the analyzers measurements, was able to distinguish the ventilation status and the specific air quality events taking place in the classroom. The same procedure applied to the data from the sensors, even though they were not calibrated beforehand, was also able to identify the same events. The good agreement between the two sets of results validates the methodology and opens up new perspectives for a massive deployment of sensors inside buildings. 297

Because of their size and price, miniature gas sensors are good candidates for long-term, large-scale, continuous monitoring of the air quality in confined environments, even in the presence of occupants. In spite of their still somewhat limited metrological performances, these tools are able to provide relevant information on the pollutants spatial and temporal evolution. They can therefore be used to identify pollution sources and automatically control ventilation and remediation systems, provided they are associated with adequate data treatment procedures. In the present study, four sensors for the detection of CO2, NO, NO2 and O3 have been deployed, without previous calibration, inside a classroom of a low energy high school building, together with standard analytical instruments. The data are analyzed with a procedure based on the bisecting K-means algorithm. This unsupervised classification allows the identification of similar measurements, which can be merged into clusters. An excellent agreement has been found between the classification results provided by the analyzers and by the sensors, even if the latter were not calibrated before deployment. These results validate the data treatment methodology proposed in this work, and demonstrate the potential of using commercial micro sensors in real conditions.

The contributions of the five mv4 Int- and two mv4 Xis arm-binding sites to the spatial intasome organization of bacteriophage mv4 were found not to be equivalent. The 8-bp overlap region was mapped to the left extremity of the core region and is directly flanked by the P2 Int arm-binding site. These results and the absence of characteristic Int core-binding sites suggest that the P2 site is the determinant for integrase positioning and recognition of the core region.

While indoor air quality issues have received increasing attention the past decades, detailed investigations of primary sources of indoor pollution are still difficult to carry out. There is a lack of analytical tools and measurement procedures to identify sources of pollutants and to characterize their emissions. Formaldehyde is a ubiquitous pollutant in indoor environments, which is known to lead to adverse health effects. This study describes a measurement procedure to apportion formaldehyde emissions from building and furnishing materials and presents a source apportionment study performed in French public schools. More than 29 sources of formaldehyde were characterized in each investigated classroom, with higher emissions from building materials compared to furnishing materials. Formaldehyde emission rates measured using passive flux samplers (PFS) range from 1.2 to 252 μg/m2/h, highlighting several strong emitters made of wood products and foam. Interestingly, the ceiling was identified as the main source of formaldehyde in most classrooms. Measured emissions and air exchange rates were constrained in a mass balance model to evaluate the impact of formaldehyde reduction strategies. These results indicate that formaldehyde concentrations can be reduced by 87–98% by removing or replacing the main source of emission by a less emissive material and by increasing the air exchange rate to 1 h−1. In addition, an intercomparison of total emissions calculated from (1) PFS measurements and from (2) measured formaldehyde concentrations and air exchange rates indicate that an unidentified sink of formaldehyde may exist in indoor environments.

The presence of β-mannosides in their cell walls confers specific features on the pathogenic yeasts Candida albicans and Candida glabrata compared with non-pathogenic yeasts. In the present study, we investigated the enzymatic properties of Bmt1 (β-mannosyltransferase 1), a member of the recently identified β-mannosyltransferase family, from C. albicans. A recombinant soluble enzyme lacking the N-terminal region was expressed as a secreted protein from the methylotrophic yeast Pichia pastoris. In parallel, functionalized natural oligosaccharides isolated from Saccharomyces cerevisiae and a C. albicans mutant strain, as well as synthetic α-oligomannosides, were prepared and used as potential acceptor substrates. Bmt1p preferentially utilizes substrates containing linear chains of α-1,2-linked mannotriose or mannotetraose. The recombinant enzyme consecuti-vely transfers two mannosyl units on to these acceptors, leading to the production of α-mannosidase-resistant oligomannosides. NMR experiments further confirmed the presence of a terminal βMan (β-1,2-linked mannose) unit in the first enzyme product. In the future, a better understanding of specific β-1,2-mannosyltransferase molecular requirements will help the design of new potential antifungal drugs.

Even though clinical, epidemiological and toxicological studies have progressively provided a better knowledge of the underlying mechanisms by which air pollution-derived particulate matter (PM) exerts its harmful health effects, further in vitro studies on relevant cell systems are still needed. Hence, aiming of getting closer to the human in vivo conditions, primary human bronchial epithelial cells derived from normal subjects (NHBE) or sensitive chronic obstructive pulmonary disease (COPD)-diseased patients (DHBE) were differentiated at the air-liquid interface. Thereafter, they were repeatedly exposed to air pollution-derived PM

Lactococcus lactis subsp. lactis biovar diacetylactis strains are used in the dairy industry for generating acetoin and notably diacetyl which imparts a high level of buttery flavor notes. A collection of domesticated and environmental strains was screened for the production of diacetyl or acetoin (D/A), and citrate fermentation. Unexpectedly, both domesticated and environmental strains produced D/A. Domesticated strains belonging to the currently named "biovar diacetylactis" metabolized citrate and produced large amounts of D/A during early growth. They harbored the citP plasmid gene encoding citrate permease and a chromosomal region citM-citI-citCDEFXG involved in citrate metabolism. In these strains, citrate consumption was identified as the major determinant of aroma production. Environmental strains, specifically UCMA5716 and A12, produced as much D/A as the CitP(+) strains, though at slightly lower rates. UCMA5716 was found to contain the citM-citI-citCDEFXG cluster but not the citP gene. A12 had neither. In these strains, production rate of D/A was linearly correlated with pyruvate synthesis rate. However, the correlation factor was strain-dependent, suggesting different modes of regulation for pyruvate rerouting towards fermentation end-products and flavors. This work highlights the genetic and metabolic differences between environmental and domesticated strains. The introduction of environmental strains into industrial processes could considerably increase the diversity of starters, enhancing the delivery of new technological properties.

While the knowledge of the underlying mechanisms by which air pollution-derived particulate matter (PM) exerts its harmful health effects is still incomplete, detailed in vitro studies are highly needed. With the aim of getting closer to the human in vivo conditions and better integrating a number of factors related to pre-existing chronic pulmonary inflammatory, we sought to develop primary cultures of normal human bronchial epithelial (NHBE) cells and chronic obstructive pulmonary disease (COPD)-diseased human bronchial epithelial (DHBE) cells, grown at the air-liquid interface. Pan-cytokeratin and MUC5AC immunostaining confirmed the specific cell-types of both these healthy and diseased cell models and showed they are closed to human bronchial epithelia. Thereafter, healthy and diseased cells were repeatedly exposed to air pollution-derived PM

The 45/47 kDa Apa, an immuno-dominant antigen secreted by Mycobacterium tuberculosis is O-mannosylated at multiple sites. Glycosylation of Apa plays a key role in colonization and invasion of the host cells by M. tuberculosis through interactions of Apa with the host immune system C-type lectins. Mycobacterium marinum (M.ma) a fish pathogen, phylogenetically close to M. tuberculosis, induces a granulomatous response with features similar to those described for M. tuberculosis in human. Although M.ma possesses an Apa homologue, its glycosylation status is unknown, and whether this represents a crucial element in the pathophysiology induced by M.ma remains to be addressed. To this aim, we have identified two concanavalin A-reactive 45/47 kDa proteins from M.ma, which have been further purified by a two-step anion exchange chromatography process. Advanced liquid chromatography-nanoESI mass spectrometry-based proteomic analyses of peptides, derived from either tryptic digestion alone or in combination with the Asp-N endoproteinase, established that M.ma Apa possesses up to seven distinct O-mannosylated sites with mainly single mannose substitutions, which can be further extended at the Ser/Thr/Pro rich region near the N-terminus. This opens the way to further studies focussing on the involvement and biological functions of Apa O-mannosylation using the M.ma/zebrafish model.

Although it was identified in the cell wall of several pathogenic mycobacteria, the biological properties of dimycolyl-diarabino-glycerol have not been documented yet. In this study an apolar glycolipid, presumably corresponding to dimycolyl-diarabino-glycerol, was purified from Mycobacterium marinum and subsequently identified as a 5-O-mycolyl-β-Araf-(1→2)-5-O-mycolyl-α-Araf-(1→1')-glycerol (designated Mma_DMAG) using a combination of nuclear magnetic resonance spectroscopy and mass spectrometry analyses. Lipid composition analysis revealed that mycolic acids were dominated by oxygenated mycolates over α-mycolates and devoid of trans-cyclopropane functions. Highly purified Mma_DMAG was used to demonstrate its immunomodulatory activity. Mma_DMAG was found to induce the secretion of proinflammatory cytokines (TNF-α, IL-8, IL-1β) in human macrophage THP-1 cells and to trigger the expression of ICAM-1 and CD40 cell surface antigens. This activation mechanism was dependent on TLR2, but not on TLR4, as demonstrated by (i) the use of neutralizing anti-TLR2 and -TLR4 antibodies and by (ii) the detection of secreted alkaline phosphatase in HEK293 cells co-transfected with the human TLR2 and secreted embryonic alkaline phosphatase reporter genes. In addition, transcriptomic analyses indicated that various genes encoding proinflammatory factors were up-regulated after exposure of THP-1 cells to Mma_DMAG. Importantly, a wealth of other regulated genes related to immune and inflammatory responses, including chemokines/cytokines and their respective receptors, adhesion molecules, and metalloproteinases, were found to be modulated by Mma_DMAG. Overall, this study suggests that DMAG may be an active cell wall glycoconjugate driving host-pathogen interactions and participating in the immunopathogenesis of mycobacterial infections.

A family of nine genes encoding proteins involved in the synthesis of β-1,2 mannose adhesins of Candida albicans has been identified. Four of these genes, BMT1-4, encode enzymes acting stepwise to add β-mannoses on to cell-wall phosphopeptidomannan (PPM). None of these acts on phospholipomannan (PLM), a glycosphingolipid member of the mannose-inositol-phosphoceramide family, which contributes with PPM to β-mannose surface expression. We show that deletion of BMT5 and BMT6 led to a dramatic reduction of PLM glycosylation and accumulation of PLM with a truncated β-oligomannoside chain, respectively. Disruptions had no effect on sphingolipid biosynthesis and on PPM β-mannosylation. β-Mannose surface expression was not affected, confirming that β-mannosylation is a process based on specificity of acceptor molecules, but liable to global regulation.

The HO2 radical is one of the most important intermediate species in atmospheric chemistry. We report on the development of a new photoreactor with first in-situ measurement of HO2 radical photostationary concentrations using continuous wave cavity ring-down spectrometry (cw-CRDS). Characterization of the actinic photon flux was carried out by NO2 actinometry. Photolysis of Cl2/methanol mixtures in air under UV light allowed the measurement of HO2 photostationary concentrations of a few 1010 molecules cm-3 with an HO2 detection limit of 1.5 × 1010 molecules cm-3 at 6638.207 cm-1. The feasibility of HO2 direct measurement in a reaction chamber is demonstrated through the measurement of the HO2 overall loss at different pressures showing the importance of HO2 diffusion and wall loss in such low pressure quartz reactor. The rate coefficient for the HO2+HO2 reaction has been measured at 6.6, 24 and 118 mbar and found to be in good agreement with the recommended value.

The kinetics of the overall reaction between OH radicals and 2,3-pentanedione (1) were studied using both direct and relative kinetic methods at laboratory temperature. The low pressure fast discharge flow experiments coupled with resonance fluorescence detection of OH provided the direct rate coefficient of (2.25 ± 0.44) × 10(-12) cm(3) molecule(-1) s(-1). The relative-rate experiments were carried out both in a collapsible Teflon chamber and a Pyrex reactor in two laboratories using different reference reactions to provide the rate coefficients of 1.95 ± 0.27, 1.95 ± 0.34, and 2.06 ± 0.34, all given in 10(-12) cm(3) molecule(-1) s(-1). The recommended value is the nonweighted average of the four determinations: k(1) (300 K) = (2.09 ± 0.38) × 10(-12) cm(3) molecule(-1) s(-1), given with 2σ accuracy. Absorption cross sections for 2,3-pentanedione were determined: the spectrum is characterized by two wide absorption bands between 220 and 450 nm. Pulsed laser photolysis at 351 nm was used and the depletion of 2,3-pentanedione (2) was measured by GC to determine the photolysis quantum yield of Φ(2) = 0.11 ± 0.02(2σ) at 300 K and 1000 mbar synthetic air. An upper limit was estimated for the effective quantum yield of 2,3-pentanedione applying fluorescent lamps with peak wavelength of 312 nm. Relationships between molecular structure and OH reactivity, as well as the atmospheric fate of 2,3-pentanedione, have been discussed.

Nucleolin is a major nucleolar protein involved in fundamental processes of ribosome biogenesis, regulation of cell proliferation and growth. Nucleolin is known to shuttle between nucleus, cytoplasm and cell surface. We have previously found that nucleolin undergoes complex N- and O-glycosylations in extra-nuclear isoforms. We found that surface nucleolin is exclusively glycosylated and that N-glycosylation is required for its expression on the cells. Interestingly, the two N-glycans are located in the RNA-binding domains (RBDs) which participate in the self-association properties of nucleolin. We hypothesized that the occupancy of RBDs by N-glycans plays a role in these self-association properties. Here, owing to the inability to quantitatively produce full-size nucleolin, we expressed four N-glycosylation nucleolin variants lacking the N-terminal acidic domain in a baculovirus/insect cell system. As assessed by heptafluorobutyrate derivatization and mass spectrometry, this strategy allowed the production of proteins bearing or not paucimannosidic-type glycans on either one or two of the potential N-glycosylation sites. Their structure was investigated by circular dichroism and fluorimetry, and their ability to self-interact was analyzed by electrophoresis and surface plasmon resonance. Our results demonstrate that all nucleolin-derived variants are able to self-interact and that N-glycosylation on both RBD1 and RBD3, or RBD3 alone, but not RBD1 alone, modifies the structure of the N-terminally truncated nucleolin and enhances its self-association properties. In contrast, N-glycosylation does not modify interaction with lactoferrin, a ligand of cell surface nucleolin. Our results suggest that the occupancy of the N-glycosylation sites may contribute to expression and functions of surface nucleolin. Structured digital abstract • NCT binds to NCT by surface plasmon resonance (View interaction) • R3CT binds to R3CT by surface plasmon resonance (View interaction) • RCT binds to RCT by surface plasmon resonance (View interaction) • RCT binds to RCT by blue native page (View interaction) • NCT binds to lactoferrin by surface plasmon resonance (View interaction) • R3CT binds to R3CT by blue native page (View interaction)

We report on the development of a new environmental simulation chamber coupled with an in situ continuous wave cavity ring-down spectrometer operating in the near IR (∼1.5 μm). The first application reported in this paper dealt with the chemical mechanism of UV photolysis of methyl nitrite (CH(3)ONO) in air. HONO has been detected for the first time and shown to be formed in the OH + CH(3)ONO reaction. A dense spectrum of cis-HONO absorption lines has been observed near 1.5 μm, in agreement with a previous study (Guilmot et al.). CH(2)O has been measured as primary product with good sensitivity and time resolution. In contrast to Zhao et al., we did not detect any NO(2) absorption features in this wavelength range. Calibration experiments provided very low NO(2) absorption cross sections in this region (∼10(-25) cm(2)), leading to conclude that NO(2) cannot be observed in this wavelength range in the presence of equal amounts of CH(2)O.

The integrase of the temperate bacteriophage mv4 catalyzes site-specific recombination between the phage attP site and the host attB site during Lactobacillus delbrueckii lysogenization. The mv4 prophage is excised during the induction of lytic growth. Excisive site-specific recombination between the attR and attL sites is also catalyzed by the phage-encoded recombinase, but the directionality of the recombination is determined by a second phage-encoded protein, the recombination directionality factor (RDF). We have identified and functionally characterized the RDF involved in site-specific excision of the prophage genome. The mv4 RDF, mv4 Xis, is encoded by the second gene of the early lytic operon. It is a basic protein of 56 amino acids. Electrophoretic mobility shift assays demonstrated that mv4 Xis binds specifically to the attP and attR sites via two DNA-binding sites, introducing a bend into the DNA. In vitro experiments and in vivo recombination assays with plasmids in Escherichia coli and Lactobacillus plantarum demonstrated that mv4 Xis is absolutely required for inter- or intramolecular recombination between the attR and attL sites. In contrast to the well-known phage site-specific recombination systems, the integrative recombination between the attP and attB sites seems not to be inhibited by the presence of mv4 Xis.

Since 1990, the MERA (MEsure des Retombees Atmospheriques, French acronym for background air pollution monitoring) network has been focused on the composition of the lower troposphere within the EMEP program. In particular, 46 non-methane hydrocarbon (NMHC) concentrations have been measured between 1997 and 2006 at three MERA sites. The analysis of temporal trends using Mann-Kendall and Sen methods showed a global decrease of anthropogenic NMHC. These results are in accordance with the trends observed on other sites in Europe and follow the decrease of VOCs emissions in France. Nevertheless the concentrations of long-life species like ethane seem to remain steady showing the growing influence of most distant source areas. In addition isoprene concentrations are typically higher in France than in other countries in Europe and slightly rising. Data analysis was performed using positive matrix factorization (PMF). Five similar PMF factors are identified as aged profiles for the three sites. The examination of factor contributions made it possible to determine a hierarchy in source influence. A higher contribution of evaporative sources was observed on the southern site while residential heating was the main factor for the other two. This work was completed by a clustering analysis (K-means) of air mass trajectories in order to apportion source contribution depending on air mass origins. Two main groups have been distinguished: (1) older and diluted air masses from an oceanic origin; and (2) anthropogenic and closer sources indicating continental influence.

Kinetics of the •OH-initiated reactions of acetic acid and its deuterated isomers have been investigated performing simulation chamber experiments at T = 300 ± 2 K. The following rate constant values have been obtained (± 1σ, in cm3 molecule−1 s−1): k 1(CH3C(O)OH + •OH) = (6.3 ± 0.9) × 10−13, k 2(CH3C(O)OD + •OH) = (1.5 ± 0.3) × 10−13, k 3(CD3C(O)OH + •OH) = (6.3 ± 0.9) × 10−13, and k 4(CD3C(O)OD + •OH) = (0.90 ± 0.1) × 10−13. This study presents the first data on k 2(CH3C(O)OD + •OH). Glyoxylic acid has been detected among the products confirming the fate of the •CH2C(O)OH radical as suggested by recent theoretical studies.

Les composes atmospheriques, qu’ils soient de sources naturelles ou anthropiques, peuvent etre transformes, transportes sur de longues distances et transferes de l’atmosphere vers un autre reservoir via les retombees atmospheriques. Ces composes, bien que presents a l’etat de trace, peuvent generer des evenements de pollution, dont la prevision est utile a la protection de la sante publique, des milieux naturels et du climat. Au cours des trois dernieres decennies, des politiques de reduction des emissions polluantes ont ete mises en place en Europe et sur d’autres continents afin de limiter leurs effets. Pour un certain nombre de ces composes, l’efficacite de ces politiques peut etre evaluee par le suivi a long terme de la chimie des retombees atmospheriques, ainsi que sur la definition de charges critiques pour un ecosysteme, definies (le milieu forestier dans cet article) comme le depot anthropique maximal qui ne conduira pas a un depassement des concentrations critiques au sein du compartiment etudie, en l’etat actuel des connaissances. En France, des observatoires nationaux (MERA, RENECOFOR/CATAENAT) sont dedies a la surveillance des retombees atmospheriques depuis les annees 90. Parmi les elements majeurs inorganiques mesures depuis l’origine, les ions sulfates (SO42-), nitrates (NO3-) et ammonium (NH4+) sont particulierement suivis pour leurs impacts acidifiant et eutrophisant sur les ecosystemes. Ces ions solubilises dans l’eau sont issus de la transformation des gaz precurseurs SOX, NOX, et NH3 emis majoritairement par les activites anthropiques. La mutualisation des mesures de ces observatoires offre un jeu de donnees inedit pour etudier l’evolution de la composition chimique des retombees atmospheriques au niveau national ou europeen et permet une comparaison avec l’evolution des emissions des polluants primaires. Les variations sur les memes periodes des cartes de charges critiques et de leurs depassements permettent de constater directement les effets des politiques de reduction des emissions mises en œuvre sur les ecosystemes terrestres.

International audience; Fungal glycosylinositolphosphoceramides (GIPCs) are involved in cell growth and fungal-host interactions. In this study, six GIPCs from the mycelium of the human pathogen Aspergillus fumigatus were purified and characterized using Q-TOF mass spectrometry and 1H, 13C, and 31P NMR. All structures have the same inositolphosphoceramide moiety with the presence of a C(18:0)-phytosphingosine conjugated to a 2-hydroxylated saturated fatty acid (2-hydroxy-lignoceric acid). The carbohydrate moiety defines two types of GIPC. The first, a mannosylated zwitterionic glycosphingolipid contains a glucosamine residue linked in alpha1-2 to an inositol ring that has been described in only two other fungal pathogens. The second type of GIPC presents an alpha-Manp-(1-->3)-alpha-Manp-(1-->2)-IPC common core. A galactofuranose residue is found in four GIPC structures, mainly at the terminal position via a beta1-2 linkage. Interestingly, this galactofuranose residue could be substituted by a choline-phosphate group, as observed only in the GIPC of Acremonium sp., a plant pathogen.

The lysogeny region of the Lactobacillus delbrueckii bacteriophage mv4 contains two divergently oriented ORFs coding for the Rep (221 aa) and Tec (64 aa) proteins. The transcription of these two genes was analysed by primer extension and Northern blot experiments on lysogenic strains. The location of the transcription initiation sites of rep and tec in the intergenic region allowed the identification of the divergently oriented non overlapping promoters P rep and P tec . Transcriptional fusions analysis showed that Rep negatively regulates the P tec promoter and activates its own transcription, and that Tec is a negative regulator of the two promoters. As demonstrated by gel mobility shift assays, the repressor Rep binds to a single specific 17 bp site located between the P tec − 10 and − 35 regions whereas Tec binds to a single specific 40 bp long complex operator site located between the two promoters. The presence of a single specific operator site for each repressor in the intergenic region is an unusual feature.

Previous studies have provided some insight into the Saharan dust deposition at a few specific locations from observations over long time periods or intensive field campaigns. However, no assessment of the dust deposition temporal variability in connection with its regional spatial distribution has been achieved so far from network observations over more than 1 year. To investigate dust deposition dynamics at the regional scale, five automatic deposition collectors named CARAGA (Collecteur Automatique de Retombées Atmosphériques insolubles à Grande Autonomie in French) have been deployed in the western Mediterranean region during 1 to 3 years depending on the station. The sites include, from south to north, Lampedusa, Majorca, Corsica, Frioul and Le Casset (southern French Alps). Deposition measurements are performed on a common weekly period at the five sites. The mean dust deposition fluxes are higher close to the northern African coasts and decrease following a south-north gradient, with values from 7.4g m year in Lampedusa (35°31'N, 12°37'E) to 1g m year in Le Casset (44°59'N, 6°28'E). The maximum deposition flux recorded is of 3.2g m wk in Majorca with only two other events showing more than 1g m wk in Lampedusa, and a maximum of 0.5g m wk in Corsica. The maximum value of 2.1g m year observed in Corsica in 2013 is much lower than existing records in the area over the 3 previous decades (11 4g m year ). From the 537 available samples, 98 major Saharan dust deposition events have been identified in the records between 2011 and 2013. Complementary observations provided by both satellite and air mass trajectories are used to identify the dust provenance areas and the transport pathways from the Sahara to the stations for the studied period. Despite the large size of African dust plumes detected by satellites, more than 80% of the major dust deposition events are recorded at only one station, suggesting that the dust provenance, transport and deposition processes (i.e. wet vs. dry) of dust are different and specific for the different deposition sites in the Mediterranean studied area. The results tend to indicate that wet deposition is the main form of deposition for mineral dust in the western Mediterranean basin, but the contribution of dry deposition (in the sense that no precipitation was detected at the surface) is far from being negligible, and contributes 10 to 46% to the major dust deposition events, depending on the sampling site. ' Author(s) 2016., This study was funded by the PRIMEQUALADEME programme on “Pollution atmosphérique longue distance” through the research project “Mesure du dépôt atmosphérique et validation de sa représentation dans les modèles régionaux” (DEMO project, contract no. 0962c0067). This project was also funded by the MISTRALS (Mediterranean Integrated Studies at Local and Regional Scales) programme as part of the Chemistry-Aerosol Mediterranean Experiment (ChArMEx) and by the Spanish Government project ChArMEx: aerosols deposition ref: CTM2011-14036-E.

Dicarbonyls in the atmosphere mainly arise from secondary sources as reaction products in the degradation of a large number of volatile organic compounds (VOC). Because of their sensitivity to solar radiation, photodissociation of dicarbonyls can dominate the fate of these VOC and impact the atmospheric radical budget. The photolysis of 2,3-pentanedione (PTD) has been investigated for the first time as a function of pressure in a static reactor equipped with continuous wave cavity ring-down spectroscopy to measure the HO2 radical photostationary concentrations along with stable species. We showed that (i) Stern-Volmer plots are consistent with low OH-radical formation yields in RCO + O2 reactions, (ii) the decrease of the photodissociation rate due to pressure increase from 26 to 1000 mbar is of about 30%, (iii) similarly to other dicarbonyls, the Stern-Volmer analysis shows a curvature at the lower pressure investigated, which may be assigned to the existence of excited singlet and triplet PTD states, (iv) PTD photolysis at 66 mbar leads to CO2, CH2O and CO with yields of (1.16 ± 0.04), (0.33 ± 0.02) and (0.070 ± 0.005), respectively, with CH2O yield independent of pressure up to 132 mbar and CO yield in agreement with that obtained at atmospheric pressure by Bouzidi et al. (2014), and (v) the PTD photolysis mechanism remains unchanged between atmospheric pressure and 66 mbar. As a part of this work, the O2 broadening coefficient for the absorption line of HO2 radicals at 6638.21 cm(-1) has been determined (γO2 = 0.0289 cm(-1) atm(-1)).

Pulsed-field gel electrophoresis (PFGE), developed in the mid-1980s, rapidly became a "gold standard" method for analyzing bacterial chromosomes. Today, although outcompeted in resolution by alternative methods, such as optical mapping, and not applicable for high-throughput analyses, PFGE remains a valuable method for bacterial strain typing. Here, we describe optimized protocols for macrorestriction fingerprinting, characterization of plasmid complements, and gene localization by DNA-DNA hybridization of Lactococcus lactis genomes.

Galactomannan is a characteristic polysaccharide of the human filamentous fungal pathogen Aspergillus fumigatus that can be used to diagnose invasive aspergillosis. In this study, we report the isolation of a galactomannan fraction associated to membrane preparations from A. fumigatus mycelium by a lipid anchor. Specific chemical and enzymatic degradations and mass spectrometry analysis showed that the lipid anchor is a glycosylphosphatidylinositol (GPI). The lipid part is an inositol phosphoceramide containing mainly C18-phytosphingosine and monohydroxylated lignoceric acid (2OH-C(24:0) fatty acid). GPI glycan is a tetramannose structure linked to a glucosamine residue: Manalpha1-2Manalpha1-2Manalpha1-6Manalpha1-4GlcN. The galactomannan polymer is linked to the GPI structure through the mannan chain. The GPI structure is a type 1, closely related to the one previously described for the GPI-anchored proteins of A. fumigatus. This is the first time that a fungal polysaccharide is shown to be GPI-anchored.

Candida albicans strains consist of serotypes A and B depending on the presence of terminal beta-1,2-linked mannose residues in the acid-stable part of serotype A phosphopeptidomannan (PPM). The distribution of C. albicans serotypes varies according to country and human host genetic and infectious backgrounds. However, these epidemiological traits have not yet been related to a phenotypically stable molecule as cell surface expression of the serotype A epitope depends on the growth conditions. We have shown that C. albicans serotype A associates beta-mannose residues with another molecule, phospholipomannan (PLM), which is a member of the mannoseinositolphosphoceramide family. In this study, PLM from serotype B strains was analysed in order to provide structural bases for the differences in molecular mass and antigenicity observed between PLMs from both serotypes. Through these analyses, carbon 10 was shown to be the location of a second hydroxylation of fatty acids previously unknown in fungal sphingolipids. Minor differences observed in the ceramide moiety appeared to be strain-dependent. More constant features of PLM from serotype B strains were the incorporation of greater amounts of phytosphingosine C20, a twofold reduced glycosylation of PLM and overexpression of a beta-1,2 mannotriose, the epitope of protective antibodies. This specific beta-mannosylation was observed even when growth conditions altered serotype A PPM-specific epitopes, confirming the potential of PLM as a phenotypically stable molecule for serotyping. This study also suggests that the regulation of beta-mannosyltransferases, which define specific immunomodulatory adhesins whose activity depends on the mannosyl chain length, are part of the genetic background that differentiates serotypes.

Pulsed-field gel electrophoresis (PFGE), developed in the mid-1980s, rapidly became a "gold standard" method for analyzing bacterial chromosomes. Today, although outcompeted in resolution by alternative methods, such as optical mapping, and not applicable for high-throughput analyses, PFGE remains a valuable method for bacterial strain typing. Here, we describe optimized protocols for macrorestriction fingerprinting, characterization of plasmid complements, and gene localization by DNA-DNA hybridization of Lactococcus lactis genomes.