Hétérotrophie algale : effets de la gentamycine et de la cycloheximide sur les activités hétérotrophes et photosynthétiques des bacteries et des algues

Afin de tenter de déterminer les parts respectives des activités bactérienne et algale, nous avons testé d'une part, l'action d'une substance antibactérienne (la gentamycine), et d'autre part, l'action d'un inhibiteur métabolique des cellules eucaryotes (la cycloheximide) sur des cultures d'algues et de bactéries et sur des échantillons provenant du milieu naturel et soumis à des filtrations différentielles. Les effets de ces inhibiteurs ont été testés au niveau des activités hétérotrophe et photosynthétique globales, mais également au niveau de l'incorporation des marqueurs radioactifs lors de la synthèse des macromolécules et des composés de faible poids moléculaire.Les résultats obtenus laissent apparaître que l'inhibition de l'activité bactérienne par la gentamyclne est significative mais non complète (pourcentage d'inhibition moyen = 67 %). De plus, l'efficacité de la gentamycine augmente avec la durée d'incubation. Par ailleurs, les effets secondaires de la gentamycine sur les activités hétérotrophe et photosynthétique d'une culture de Melosira italica subsp. subarctica sont acceptables seulement pour des incubations de courte durée (< 4 heures).En revanche, l'emploi de la cycloheximide s'est révélé sans aucun effet significatif sur les activités photosynthétique et hérérotrophe de la culture de Melosira, même après 24 h d'incubation.A partir des échantillons prélevés en milieu naturel, l'emploi de la gentamycine a permis de réduire l'interférence bactérienne dans les mesures d'activité hétérotrophe algale. Enfin, nous avons pu constater que la gentamycine modifie l'allocation des marqueurs radioactifs dans les macromolécules.The ability of many planktonic algae to use particulate and/or dissolved organic carbon directly by phagotrophy or osmotrophy in laboratory cultures is well documented (DROOP, 1974; NEILSON and LEWIN, 1974; HELLEBUST and LEWIN, 1977; BIRD and KALFF, 1986). In axenic cultures, numerous microalgae grow in the dark with micromolar concentrations of diverse organic nutrients as their sole sources of carbon and energy (RIVKIN and PUTT, 1987). However, to demonstrate algal heterotrophy in the field, it is necessary to differentiate between bacterial and algal activities. In the course of this study, we tested the effect of an antibacterial substance (gentamycin) and of a metabolic inhibitor of eukaryotic cells (cycloheximide) on algal and bacterial cultures, and also on lake water samples submitted to differential filtration. The effect of these inhibitors was tested both at the overall heterotrophic and photosynthetic activities level and the level of the incorporation of radiolabeled tracers in macromolecules and low molecular weight compounds.Gentamycin was tested on bacteria and on an axenic culture of the diatom Melosira italica subsp. subarctica, the dominant species of the spring phyto-planktonic bloom of many temperate lakes. Bacterial culture was obtained by filtration of a senescent culture of Melosira through a 0.45 µm pore-size membrane. During exponential growth, gentamycin (40 µg.ml-1) was added to different flasks containing 100 ml of culture, 30 mn after gentamycin addition, Na H14CO3 (12µCi/100 ml) was introduced into the flasks. In each case, two replicates were incubated in the light and two in the dark for 0.5, 1, 2, 4, 6, 8, 12 and 24 hours. After incubation, aliquots were collected on a filtration membrane (0.45 µm) and the radioactivity was measured using an LKB liquid scintillation counter to estimate photosynthetic activity. Incorporation of inorganic carbon into macromolecules was measured using the procedure described by LANCELOT and MATHOT (1985), which allows the separation of lipids, polysaccharides, proteins and low molecular weight compounds (e.g. amino acids, organic acids and monosaccharides) by virtue of their relative solubilities in different extraction solvents :- lipids were extracted with a 2/1 (v/v) chloroform-methanol mixture;- low molecular weight compounds were extracted with hot ethanol;- proteins were precipated with TCA at elevated temperature; this also separated them from polysaccharides. Nucleic acids were recovered with the polysaccharides fraction. Results are given as a percentage of total radioactivity. The same method, as that described for photosynthetic incorporation, was used to measure incorporation of glucose 3H (19 nmoles.l-1) and for the allocation of radioactive marker in macromolecules. To assess the effect of cycloheximide, the same procedure as that described for gentamycin was used. However, due to the lack of references, three cycloheximide concentrations were tested = 50, 100 and 150 µg.ml-1, and aliquots were collected on a 0.2 µm pore-size membrane.Field samples were taken in Lake Pavin, an oligomesotrophic French lake, during the sedimentation of Melosira italica subsp. subarctica, the prevailing species of the spring bloom. The effects of gentamycin on photosynthetic and heterotrophic incorporations were tested with the came procedure as that described for cultures. However, radioactivity was measured for different size tractions : 0.2-0.45 µm, 0.45-5 µm and 5-160 µm, after incubations for 0.5, 1, 3, 6, 12 and 24 hours. Phytoplanktonic cells were counted on a Wild M40 inverted microscope and bacterial enumeration was realized in epifluorescence microscopy after staining with acridine orange (HOBBIE el al., 1977).Results show that gentamycin used at 40 µg.ml-1 inhibits significantly but not completely the activity of the bacterial culture (mean inhibition percent = 67 %). Gentamycin efficiency increases with incubation time, the inhibition reaching 81 % after 24 h. At the same time, the secondary effects of gentamycin on heterotrophic and photosynthetic activities of Melosira italica subsp. subarctica in culture were only tolerable with short incubation times (< 4 h), when the percents of inhibition were respectively 13.6 and 12.2%. On the other band, cycloheximide produced no significant effect on photosynthetic and heterotrophic activities of Melosira italica in culture, the percent of inhibition always remaining below 6.5 %.The use of gentamycin in natural samples reduced bacterial interference with algal heterotrophic activily measurements. The percent of inhibition caused by gentamycin was high (< 76 %) In the small-size fraction where the bacterial biomass predominate on the phytoplanktonic one.Lastly, in all samples, we could demonstrate that gentamycin modifies the allocation of inorganic carbon and radioactivity in macromolecules. The incorporation in proteins is significantly reduced essentially to benefit of the incorporation in low molecular weight compounds

Stratigraphy and age of the Cappadocia ignimbrites, Turkey: reconciling field constraints with paleontologic, radiochronologic, geochemical and paleomagnetic data

The stratigraphy and age of the Neogene Cappadocia ignimbrites (Central Turkey) have been inferred in previous studies from fieldwork and K–Ar age determinations. The resulting stratigraphic schemes, however, differed from each other, suggesting that further studies were required to produce a reliable succession. In this paper, we examine the chronostratigraphy of mammalian remains recovered in the continental sediments interbedded with the Cappadocia ignimbrites. Using recent advances in mammalian chronostratigraphy, we evaluate selected taxa and faunal associations to place new and independent constraints on the ignimbrite ages. The biostratigraphically bracketed ages concur with some published radiometric dates, but they disagree with others, principally at localities where major stratigraphic discrepancies have arisen in the literature. In order to reconcile these apparent inconsistencies, we combine, at selected sites, our field observations with the biostratigraphic and radiometric age limits, and we compare these with the available geochemical and magnetic data. This allows us to present revised age estimates, and a revised stratigraphy which includes the correlation of the local Sofular deposits with the large-volume Gördeles ignimbrite. The issues faced in this study apply to other ignimbrite provinces in the world. For instance, ignimbrite eruption frequency in Cappadocia is higher than the resolution of many published K–Ar ages. Furthermore, different K–Ar ages have led to the description of individual and distinct ignimbrites that fieldwork and geochemical data allow to merge into a single ignimbrite. Argon loss from pumice samples leading to radiometric “rejuvenation” provides a likely explanation for most stratigraphic discrepancies. Cappadocia is the only documented ignimbrite field in which the chronostratigraphy of vertebrate remains provides better constraints on some ignimbrite ages than scattered K–Ar dates. We further argue that K–Ar dates from the Cappadocia area are too imprecise to establish a reliable magnetostratigraphic scheme for the ignimbrite succession, with the exception of the not, vert, similar2.8 Ma Valibaba Tepe ignimbrite

The reflectivity of relativistic ultra-thin electron layers

The coherent reflectivity of a dense, relativistic, ultra-thin electron layer is derived analytically for an obliquely incident probe beam. Results are obtained by two-fold Lorentz transformation. For the analytical treatment, a plane uniform electron layer is considered. All electrons move with uniform velocity under an angle to the normal direction of the plane; such electron motion corresponds to laser acceleration by direct action of the laser fields, as it is described in a companion paper. Electron density is chosen high enough to ensure that many electrons reside in a volume \lambda_R^3, where \lambda_R is the wavelength of the reflected light in the rest frame of the layer. Under these conditions, the probe light is back-scattered coherently and is directed close to the layer normal rather than the direction of electron velocity. An important consequence is that the Doppler shift is governed by \gamma_x=(1-(V_x/c)^2)^{-1/2} derived from the electron velocity component V_x in normal direction rather than the full \gamma-factor of the layer electrons.Comment: 7 pages, 4 figures, submitted to the special issue "Fundamental Physics with Ultra-High Fields" in The European Physical Journal

Giant half-cycle attosecond pulses

Half-cycle picosecond pulses have been produced from thin photo-conductors, when applying an electric field across the surface and switching on conduction by a short laser pulse. Then the transverse current in the wafer plane emits half-cycle pulses in normal direction, and pulses of 500 fs duration and 1e6 V/m peak electric field have been observed. Here we show that single half-cycle pulses of 50 as duration and up to 1e13 V/m can be produced when irradiating a double foil target by intense few-cycle laser pulses. Focused onto an ultra-thin foil, all electrons are blown out, forming a uniform sheet of relativistic electrons. A second layer, placed at some distance behind, reflects the drive beam, but lets electrons pass straight. Under oblique incidence, beam reflection provides the transverse current, which emits intense half-cycle pulses. Such a pulse may completely ionize even heavier atoms. New types of attosecond pump-probe experiments will become possible.Comment: 5 pages, 4 figures, to be presented at LEI2011-Light at Extreme Intensities and China-Germany Symposium on Laser Acceleratio

Localization of a 64-kDa phosphoprotein in the lumen between the outer and inner envelopes of pea chloroplasts

The identification and localization of a marker protein for the intermembrane space between the outer and inner chloroplast envelopes is described. This 64-kDa protein is very rapidly labeled by [γ-32P]ATP at very low (30 nM) ATP concentrations and the phosphoryl group exhibits a high turnover rate. It was possible to establish the presence of the 64-kDa protein in this plastid compartment by using different chloroplast envelope separation and isolation techniques. In addition comparison of labeling kinetics by intact and hypotonically lysed pea chloroplasts support the localization of the 64-kDa protein in the intermembrane space. The 64-kDa protein was present and could be labeled in mixed envelope membranes isolated from hypotonically lysed plastids. Mixed envelope membranes incorporated high amounts of 32P from [γ-32P]ATP into the 64-kDa protein, whereas separated outer and inner envelope membranes did not show significant phosphorylation of this protein. Water/Triton X-114 phase partitioning demonstrated that the 64-kDa protein is a hydrophilic polypeptide. These findings suggest that the 64-kDa protein is a soluble protein trapped in the space between the inner and outer envelope membranes. After sonication of mixed envelope membranes, the 64-kDa protein was no longer present in the membrane fraction, but could be found in the supernatant after a 110000 × g centrifugation

Circulating adrenomedullin estimates survival and reversibility of organ failure in sepsis: the prospective observational multinational Adrenomedullin and Outcome in Sepsis and Septic Shock-1 (AdrenOSS-1) study

Background: Adrenomedullin (ADM) regulates vascular tone and endothelial permeability during sepsis. Levels of circulating biologically active ADM (bio-ADM) show an inverse relationship with blood pressure and a direct relationship with vasopressor requirement. In the present prospective observational multinational Adrenomedullin and Outcome in Sepsis and Septic Shock 1 (, AdrenOSS-1) study, we assessed relationships between circulating bio-ADM during the initial intensive care unit (ICU) stay and short-term outcome in order to eventually design a biomarker-guided randomized controlled trial. Methods: AdrenOSS-1 was a prospective observational multinational study. The primary outcome was 28-day mortality. Secondary outcomes included organ failure as defined by Sequential Organ Failure Assessment (SOFA) score, organ support with focus on vasopressor/inotropic use, and need for renal replacement therapy. AdrenOSS-1 included 583 patients admitted to the ICU with sepsis or septic shock. Results: Circulating bio-ADM levels were measured upon admission and at day 2. Median bio-ADM concentration upon admission was 80.5 pg/ml [IQR 41.5-148.1 pg/ml]. Initial SOFA score was 7 [IQR 5-10], and 28-day mortality was 22%. We found marked associations between bio-ADM upon admission and 28-day mortality (unadjusted standardized HR 2.3 [CI 1.9-2.9]; adjusted HR 1.6 [CI 1.1-2.5]) and between bio-ADM levels and SOFA score (p &lt; 0.0001). Need of vasopressor/inotrope, renal replacement therapy, and positive fluid balance were more prevalent in patients with a bio-ADM &gt; 70 pg/ml upon admission than in those with bio-ADM ≤ 70 pg/ml. In patients with bio-ADM &gt; 70 pg/ml upon admission, decrease in bio-ADM below 70 pg/ml at day 2 was associated with recovery of organ function at day 7 and better 28-day outcome (9.5% mortality). By contrast, persistently elevated bio-ADM at day 2 was associated with prolonged organ dysfunction and high 28-day mortality (38.1% mortality, HR 4.9, 95% CI 2.5-9.8). Conclusions: AdrenOSS-1 shows that early levels and rapid changes in bio-ADM estimate short-term outcome in sepsis and septic shock. These data are the backbone of the design of the biomarker-guided AdrenOSS-2 trial. Trial registration: ClinicalTrials.gov, NCT02393781. Registered on March 19, 2015

On Classical Solutions of 4d Supersymmetric Higher Spin Theory

We present a simple construction of solutions to the supersymmetric higher spin theory based on solutions to bosonic theories. We illustrate this for the case of the Didenko-Vasiliev solution in arXiv:0906.3898, for which we have found a striking simplification where the higher-spin connection takes the vacuum value. Studying these solutions further, we check under which conditions they preserve some supersymmetry in the bulk, and when they are compatible with the boundary conditions conjectured to be dual to certain 3d SUSY Chern-Simons-matter theories. We perform the analysis for a variety of theories with $\mathcal{N}$ = 2, $\mathcal{N}$ = 3, $\mathcal{N}$ = 4 and $\mathcal{N}$ = 6 and find a rich spectrum of $1/4$, $1/3$ and $1/2$-BPS solutions.Comment: v2: 42 pages, section 5 extended, reference added. Version to appear in JHE

New engineering approach for the development and demonstration of a multi-purpose platform for the Blue Growth Economy

Aquaculture is currently the fastest growing food sector in the world and the open oceans are seen as one of the most likely areas for large scale expansion [1], [2], [3]. The global demand for seafood is continuing to rise sharply, driven by both population growth and increased per capita consumption, whilst wild capture fisheries are constrained in their potential to produce more seafood. A recently funded EC project, the Blue Growth Farm BGF (GA n. 774426, 1st June 2018 ÷ 30th September 2021) aims at contributing to this world need with an original solution. The Blue Growth Farm proposes an efficient, cost competitive and environmentally friendly multi purpose offshore farm concept based on a modular floating structure, moored to the seabed, meeting requirements of efficiency, cost-competitiveness and environmental friendless, where automated aquaculture and renewable energy production systems are integrated and engineered for profitable applications in the open sea. In the present paper, the overall engineering approach developed to carry out the research work is presented, described and justified. Different technical and scientific challenges are addressed through an integrated industrial engineering design approach, where all disciplines are tuned to achieve the Blue Growth Farm main targets, represented by: i) guaranteeing expected nominal fish production thanks to advanced automation and remote control capabilities; ii) minimizing the pollution introduced at marine ecosystem level when exploiting the marine natural resources, whilst increasing the social acceptance and users community agreement; iii) maximizing the electricity production in the Blue Growth Farm potential installation area ecosystem to provide energy supply to the on board electrical equipment and to dispatch the extra produced electric energy to the land network. Preliminary engineering design results are promising to demonstrate effective increase of safety and efficiency by reducing on board human effort and consequently risks at offshore, thus to make commercial scale open ocean farming a reality