Toxicity, transfer and depuration of anatoxin-a (cyanobacterial neurotoxin) in medaka fish exposed by single-dose gavage

International audienceThe proliferations of cyanobacteria are increasingly prevalent in many rivers and water bodies due especially to eutrophication. This work aims to study in female medaka fish the toxicity, the transfer and the depuration of the anatoxin-a, a neurotoxin produced by benthic cyanobacterial biofilms. This work will provide answers regarding acute toxicity induced by single gavage by anatoxin-a and to the risks of exposure by ingestion of contaminated fish flesh, considering that data on these aspects remain particularly limited.The oral LD50 and NOAEL of a single dose of (±)-anatoxin-a were determined at 11.50 and 6.67 μg.g-1, respectively. Subsequently, the toxico-kinetics of the (±)-anatoxin-a was observed in the guts, the livers and the muscles of female medaka fish for 10 days. Anatoxin-a was quantified by high-resolution qTOF mass spectrometry coupled upstream to a UHPLC chromatographic chain. The toxin could not be detected in the liver after 12 h, and in the gut and muscle after 3 days. Overall, the medaka fish do not appear to accumulate (±)-anatoxin-a and to largely recover after 24 h following a single sub-acute oral liquid exposure at the NOAEL

Principal Component Analysis of Molecular Clouds: Can CO reveal the dynamics?

We use Principal Component Analysis (PCA) to study the gas dynamics in numerical simulations of typical MCs. Our simulations account for the non-isothermal nature of the gas and include a simplified treatment of the time-dependent gas chemistry. We model the CO line emission in a post-processing step using a 3D radiative transfer code. We consider mean number densities n_0 = 30, 100, 300 cm^{-3} that span the range of values typical for MCs in the solar neighbourhood and investigate the slope \alpha_{PCA} of the pseudo structure function computed by PCA for several components: the total density, H2 density, 12CO density, 12CO J = 1 -> 0 intensity and 13CO J = 1 -> 0 intensity. We estimate power-law indices \alpha_{PCA} for different chemical species that range from 0.5 to 0.9, in good agreement with observations, and demonstrate that optical depth effects can influence the PCA. We show that when the PCA succeeds, the combination of chemical inhomogeneity and radiative transfer effects can influence the observed PCA slopes by as much as ~ +/- 0.1. The method can fail if the CO distribution is very intermittent, e.g. in low-density clouds where CO is confined to small fragments.Comment: 12 pages, 8 figures, accepted for publication in MNRA

Laser femtoseconde à fibre optique émettant dans l'infrarouge moyen

Le développement de lasers émettant dans l’infrarouge moyen, une région spectrale où l’onretrouve les résonances fondamentales de la plupart des molécules sur terre, est certainementl’une des avenues les plus prometteuses en science laser. La démocratisation de ce type delaser est toutefois nécessaire afin d’exploiter leur énorme potentiel applicatif en santé, enenvironnement, en industrie et en recherche. En raison de leur simplicité, leur robustesse, leurfiabilité et leurs performances optiques inégalées, les lasers à base de fibres optiques sont parmiles candidats les mieux adaptés pour réaliser ce virage technologique vers l’infrarouge moyen.Dans le cadre de ce projet de doctorat, le premier laser à fibre optique émettant des impulsionsfemtoseconde dans l’infrarouge moyen a été réalisé. Ce laser à grande valeur ajoutée offredes propriétés temporelles, spectrales et spatiales exceptionnelles qui ouvrent la voie à denombreuses applications en spectroscopie et en interaction laser-matière.La conception, l’optimisation et l’étude théorique du laser femtoseconde émettant à 2.8μm sontprésentées aux chapitres 1 et 2 ainsi qu’à l’annexe B. La génération d’impulsions ultrabrèvesavec ce laser est basée sur la synchronisation modale passive par rotation non linéaire de lapolarisation dans une fibre de verre fluoré dopée à l’erbium. Des impulsions de durée inférieureà 300 fs, dont la puissance crête estimée est supérieure à 20 kW, ont été obtenues à partir decet oscillateur.La deuxième partie de cette thèse (chapitres 3 et 4) présente la conception et l’étude théoriquepar simulations numériques d’un amplificateur à fibre optique externe permettant d’améliorerles performances de l’oscillateur présenté dans les chapitres précédents. Cet amplificateuremployant un seul segment de fibre de verre fluoré dopé à l’erbium permet d’amplifier lesimpulsions à des puissances crêtes de plus de 200 kW. Grâce au processus d’auto décalagespectral du soliton dans cet amplificateur, la longueur d’onde centrale des impulsions amplifiéespeut être accordée de 2.8 à 3.6μm. Ce système laser femtoseconde accordable dans l’infrarougemoyen, qui constitue une avancée majeure, peut être simplifié et amélioré davantage en plusd’être adapté à plusieurs autres transitions laser dans l’infrarouge moyen. Ces travaux ouvrentla voie à l’émergence de nouvelles applications autrefois difficilement réalisables en rechercheet en industrie.The development of lasers emitting in the mid-infrared, a spectral region where the funda-mental resonances of several molecules are found, is one of the most promising avenues in laserscience. However, the democratization of these coherent light sources is required for enablingbreakthrough applications in healthcare, environment, industry and research. Thanks to theirsimplicity, robustness, reliability and their unequaled optical performances, fiber lasers are thecandidates of choice for enabling this technological shift toward the mid-infrared.In this project, the first femtosecond fiber laser emitting in the mid-infrared was realized. Thishigh added-value laser offers exceptionnal temporal, spectral and spatial properties that pavethe way for several applications in spectroscopy and laser-matter interaction.The design, optimization and theoretical study of this femtosecond laser cavity emitting at 2.8μm are presented in chapters 1 and 2 as well as in appendix B. The generation of ultrashortpulses in this laser relies on a mode-locking technique based on nonlinear polarization evolutionof the signal propagating inside an erbium-doped fluoride fiber. Pulses with durations below300 fs and estimated peak powers above 20 kW were directly generated from this oscillator.The second part of this thesis (chapters 3 and 4) presents the design and numerical studyof an external fiber amplifier that significantly improves the performances of the oscillator.This amplifier that uses only one segment of erbium-doped fluorozirconate fiber enables thegeneration of ultrashort pulses with peak powers above 200 kW. Due to the soliton self-frequency shift process occuring inside the amplifier, the central wavelength of the outputpulses can be tuned from 2.8 to 3.6μm. This tunable femtosecond system in the mid-infraredcan be further simplified and improved and can also be adapted to other novel laser transitionsin the mid-infrared. This work paves the way for new applications to emerge both in thescientific and the industrial worlds

Lasers à fibre femtoseconde utilisant une paire de réseaux de Bragg à pas variable

Ce mémoire traite de la conception et de la mise en opération d’un nouveau type de laser femtoseconde à fibre optique intégrant une paire de réseaux de Bragg à pas variable. La présence de ces éléments de dispersion opposée dans une cavité en anneau révèle une toute nouvelle dynamique temporelle en régime permanent. Une impulsion femtoseconde qui se propage dans une section de la cavité est localement transformée en une impulsion picoseconde grandement chirpée dans l’autre section. Cette dernière section agit essentiellement comme une ligne à délai purement dispersive et peut donc être modifiée de façon à varier la dispersion nette ainsi que la cadence du laser sans pour autant accroître les effets non linéaires. Le laser à l’erbium introduit dans ce mémoire, qui génère des impulsions sous la centaine de femtosecondes dans tous les régimes de dispersion étudiés, pourrait éventuellement devenir une source à impulsions ultrabrèves (< 50 fs) de très grande énergie (> 20 nJ). Cette source serait donc une excellente alternative tout-fibre aux lasers à l’état solide femtoseconde.In this master’s thesis, we present a new type of femtosecond fiber ring laser that uses a pair of chirped fiber Bragg gratings with opposite dispersion. The presence of such elements in a ring cavity reveals a new mode locking regime where a femtosecond pulse evolving in one section of the cavity is locally transformed into a highly chirped picosecond pulse that propagates in the remaining part of the cavity. The section in which the highly chirped pulse propagates acts essentially as an all-fiber linear dispersive delay line. This portion can thus be modified in order to change the net cavity dispersion or the repetition rate of the laser without significantly increasing the nonlinear effects in the cavity. This erbium-doped fiber laser that generates sub-100 fs pulses in any dispersion regime can potentially produce highenergy ultrashort pulses (> 20 nJ; < 50 fs). This source appears to be a practical all-fiber alternative to femtosecond solid-state lasers

Electron Transfer Precedes ATP Hydrolysis during Nitrogenase Catalysis

The biological reduction of N2 to NH3 catalyzed by Mo-dependent nitrogenase requires at least eight rounds of a complex cycle of events associated with ATP-driven electron transfer (ET) from the Fe protein to the catalytic MoFe protein, with each ET coupled to the hydrolysis of two ATP molecules. Although steps within this cycle have been studied for decades, the nature of the coupling between ATP hydrolysis and ET, in particular the order of ET and ATP hydrolysis, has been elusive. Here, we have measured first-order rate constants for each key step in the reaction sequence, including direct measurement of the ATP hydrolysis rate constant: kATP = 70 s−1, 25 °C. Comparison of the rate constants establishes that the reaction sequence involves four sequential steps: (i) conformationally gated ET (kET = 140 s−1, 25 °C), (ii) ATP hydrolysis (kATP = 70 s−1, 25 °C), (iii) Phosphate release (kPi = 16 s−1, 25 °C), and (iv) Fe protein dissociation from the MoFe protein (kdiss = 6 s−1, 25 °C). These findings allow completion of the thermodynamic cycle undergone by the Fe protein, showing that the energy of ATP binding and protein–protein association drive ET, with subsequent ATP hydrolysis and Pi release causing dissociation of the complex between the Feox(ADP)2 protein and the reduced MoFe protein

Arsenics as bioenergetic substrates

AbstractAlthough at low concentrations, arsenic commonly occurs naturally as a local geological constituent. Whereas both arsenate and arsenite are strongly toxic to life, a number of prokaryotes use these compounds as electron acceptors or donors, respectively, for bioenergetic purposes via respiratory arsenate reductase, arsenite oxidase and alternative arsenite oxidase. The recent burst in discovered arsenite oxidizing and arsenate respiring microbes suggests the arsenic bioenergetic metabolisms to be anything but exotic. The first goal of the present review is to bring to light the widespread distribution and diversity of these metabolizing pathways. The second goal is to present an evolutionary analysis of these diverse energetic pathways. Taking into account not only the available data on the arsenic metabolizing enzymes and their phylogenetical relatives but also the palaeogeochemical records, we propose a crucial role of arsenite oxidation via arsenite oxidase in primordial life. This article is part of a Special Issue entitled: The evolutionary aspects of bioenergetic systems

Enzyme phylogenies as markers for the oxidation state of the environment: The case of respiratory arsenate reductase and related enzymes

<p>Abstract</p> <p>Background</p> <p>Phylogenies of certain bioenergetic enzymes have proved to be useful tools for deducing evolutionary ancestry of bioenergetic pathways and their relationship to geochemical parameters of the environment. Our previous phylogenetic analysis of arsenite oxidase, the molybdopterin enzyme responsible for the biological oxidation of arsenite to arsenate, indicated its probable emergence prior to the Archaea/Bacteria split more than 3 billion years ago, in line with the geochemical fact that arsenite was present in biological habitats on the early Earth. Respiratory arsenate reductase (Arr), another molybdopterin enzyme involved in microbial arsenic metabolism, serves as terminal oxidase, and is thus situated at the opposite end of bioenergetic electron transfer chains as compared to arsenite oxidase. The evolutionary history of the Arr-enzyme has not been studied in detail so far.</p> <p>Results</p> <p>We performed a genomic search of genes related to <it>arrA </it>coding for the molybdopterin subunit. The multiple alignment of the retrieved sequences served to reconstruct a neighbor-joining phylogeny of Arr and closely related enzymes. Our analysis confirmed the previously proposed proximity of Arr to the cluster of polysulfide/thiosulfate reductases but also unravels a hitherto unrecognized clade even more closely related to Arr. The obtained phylogeny strongly suggests that Arr originated after the Bacteria/Archaea divergence in the domain Bacteria, and was subsequently laterally distributed within this domain. It further more indicates that, as a result of accumulation of arsenate in the environment, an enzyme related to polysulfide reductase and not to arsenite oxidase has evolved into Arr.</p> <p>Conclusion</p> <p>These findings are paleogeochemically rationalized by the fact that the accumulation of arsenate over arsenite required the increase in oxidation state of the environment brought about by oxygenic photosynthesis.</p

Intermolecular electron transfer in two-iron superoxide reductase: a putative role for the desulforedoxin center as an electron donor to the iron active site.

International audienceSuperoxide reductase (SOR) is a superoxide detoxification system present in some microorganisms. Its active site consists of an unusual mononuclear iron center with an FeN4S1 coordination which catalyzes the one-electron reduction of superoxide to form hydrogen peroxide. Different classes of SORs have been described depending on the presence of an additional rubredoxin-like, desulforedoxin iron center, whose function has remained unknown until now. In this work, we investigated the mechanism of the reduction of the SOR iron active site using the NADPH:flavodoxin oxidoreductase from Escherichia coli, which was previously shown to efficiently transfer electrons to the Desulfoarculus baarsii SOR. When present, the additional rubredoxin-like iron center could function as an electronic relay between cellular reductases and the iron active site for superoxide reduction. This electron transfer was mainly intermolecular, between the rubredoxin-like iron center of one SOR and the iron active site of another SOR. These data provide the first experimental evidence for a possible role of the rubredoxin-like iron center in the superoxide detoxifying activity of SOR

Physical Properties and Galactic Distribution of Molecular Clouds identified in the Galactic Ring Survey

We derive the physical properties of 580 molecular clouds based on their 12CO and 13CO line emission detected in the University of Massachusetts-Stony Brook (UMSB) and Galactic Ring surveys. We provide a range of values of the physical properties of molecular clouds, and find a power-law correlation between their radii and masses, suggesting that the fractal dimension of the ISM is around 2.36. This relation, M = (228 +/- 18) R^{2.36+/-0.04}, allows us to derive masses for an additional 170 GRS molecular clouds not covered by the UMSB survey. We derive the Galactic surface mass density of molecular gas and examine its spatial variations throughout the Galaxy. We find that the azimuthally averaged Galactic surface density of molecular gas peaks between Galactocentric radii of 4 and 5 kpc. Although the Perseus arm is not detected in molecular gas, the Galactic surface density of molecular gas is enhanced along the positions of the Scutum-Crux and Sagittarius arms. This may indicate that molecular clouds form in spiral arms and are disrupted in the inter-arm space. Last, we find that the CO excitation temperature of molecular clouds decreases away from the Galactic center, suggesting a possible decline in the star formation rate with Galactocentric radius. There is a marginally significant enhancement in the CO excitation temperature of molecular clouds at a Galactocentric radius of about 6 kpc, which in the longitude range of the GRS corresponds to the Sagittarius arm. This temperature increase could be associated with massive star formation in the Sagittarius spiral arm