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International audience; The widespread pollution from the release of microfibers is an emerging concern as they are a potential threat to the environment. Their identification in samples in terms of quantity and pathways remain a challenge as contamination can be a major source of error. A systematic study of synthetic microfibers (MFs) has been carried out in different environmental compartments of an urban area and in the surface waters of the northwestern Mediterranean.The quantity, size and type of polymer of MFs were recorded in air, in waste water from a domestic washing machine, at the inlet and outlet of the Haliotis urban wastewater treatment plant (WWTP) in Nice (Provence Alpes Côte-d'Azur, France) and in a variety of coastal and offshore areas. The results showed that MFs released by clothes during washing (on average of 13 × 106 MFs per m3) are an important emitter of microplastics. Despite its high removal efficiency (87.5% to 98.5%) by Haliotis, a large number of MFs, estimated at 4.3 billion, enter the marine environment daily from the treatment plant. The attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR) characterization of the raw materials showed that 14 to 50% of fibers are synthetic, mostly polyester and polyamide, the remaining 35 to 72% being natural polymers (cotton, wool) or manufactured by processing natural polymers (especially cellulose). MFs were found in all environmental compartments studied and appear to be widespread in coastal and offshore surface waters with concentrations varying from 2.6 × 103 to 3.70 × 104 m−3. The sources of MFs in the marine environment are multiple, with laundry fibers discharges from WWTP and the atmospheric transport of urban fibers are among the main pathways.

In this study, the accuracy and the precision corresponding to Li isotopic measurements of low level samples such as marine and coastal carbonates are estimated. To this end, a total of fifty‐four analyses of a Li‐pure reference material (Li7‐N) at concentrations ranging from 1 to 6 ng ml−1 were first performed. The average δ7Li values obtained for solutions with and without chemical purification were 30.3 ± 0.4‰ (2s, n = 19) and 30.2 ± 0.4‰ (2s, n = 36), respectively. These results show that the chosen Li chemical extraction and purification procedure did not induce any significant isotope bias. Two available carbonate reference materials (JCt‐1 and JCp‐1) were analysed, yielding mean δ7Li values of 18.0 ± 0.27‰ (2s, n = 6) and 18.8 ± 1.8‰ (2s, n = 9), respectively. Small powder aliquots ( 2), contamination from silicates cannot be avoided, even at low HCl molarity (⪡ 0.1 mol l−1). Finally, bulk carbonate and foraminifera extracted from the same core sample exhibited significant discrepancies: δ7Li values of foraminifera were more reproducible but were significantly lower. They were also associated with lower Sr/Ca and higher Mn/Ca ratios, suggesting a higher sensitivity to diagenesis, although specific vital effects cannot be fully ruled out.

We have investigated provenance and weathering proxies of the clay-size sediment exported from the Nile River basin over the last 110,000 years. Using neodymium isotope composition of sediments from both the Nile Deep Sea-Fan and Lake Tana, we show that the Nile River branches draining the Ethiopian Highlands have remained the main contributors of clays to the Nile delta during the Late Quaternary. We demonstrate that fluctuations of clay-size particle contribution to the Nile Delta are mainly driven by orbital precession cycle, which controls summer insolation and consequently the African monsoon intensity changes. Our results indicate that - over the last 110,000 years – the proportion of clays coming from Ethiopian Traps fluctuates accordingly to the intensity of the last 5 precession cycles (MIS 5 to MIS 1). However, there is a threshold effect in the transport efficiency during the lowest insolation minima (arid periods), in particular during the MIS3. Several arid events corresponding to the Heinrich Stadial periods are associated with small or negligible clay source changes while chemical weathering proxies, such as δ7Li, Mg/Ti and K/Ti, vary significantly. This suggests a straightforward control of weathering by hydro-climate changes over centennial to millennial timescales. Our data also suggests a significant but more progressive influence of the temperature decrease between 110kyr and 20kyr. Taken altogether, the observed tight coupling between past climate variations and silicate weathering proxies leads us to conclude that precipitation changes in northeast Africa can impact soil development over a few hundred years only, while the influence of temperature appears more gradual.

The dissolution of anthropogenic atmospheric CO 2 in seawater is suspected to exert a strong pressure on biological processes as well as on carbon and nutrient cycles. Transparent exopolymeric particles (TEP) are polysaccharide particles, formed by aggregation of polymers exuded by phytoplankton and strongly involved in organic matter sedimentation. A mesocosm experiment was performed from February to March 2013 in the coastal waters of the Northwestern Mediterranean Sea, a region characterised by low-nutrient low-chlorophyll (LNLC) levels. We aimed to determine the effect of ocean acidification on the organic carbon pool of TEP produced by a natural phytoplankton community. The experiment was conducted in nine mesocosms of 50 m 3 deployed for 12 days, and subjected to seven partial pressures of CO 2 ( p CO 2 ) levels: one control level in triplicate and six elevated levels between 450 and 1250 μatm. The use of different analytical methods allowed the assessment of TEP density, volume concentration and size distribution as well as both TEP and TEP precursors carbon content. TEP contributed vastly to the particulate organic carbon pool (∼62%), and were mainly produced by small-sized phytoplankton such as pico- and nanophytoplankton. TEP precursors carbon content represented three times the carbon content of particulate TEP, showing that this pool has to be considered in experiments focused on the environmental control of TEP production. There was no evidence that TEP and TEP precursors were dependent on p CO 2 . These parameters exhibited clear temporal dynamics, with tight links to community composition, nutrient availability and other environmental parameters.

International audience; The development of methods to automatically determine the chemical nature of microplastics by FTIR-ATR spectra is an important challenge. A machine learning method, named k-nearest neighbors classification, has been applied on spectra of microplastics collected during Tara Expedition in the Mediterranean Sea (2014). To realize these tests, a learning database composed of 969 microplastic spectra has been created. Results show that the machine learning process is very efficient to identify spectra of classical polymers such as poly(ethylene), but also that the learning database must be enhanced with less common microplastic spectra. Finally, this method has been applied on more than 4000 spectra of unidentified microplastics. The verification protocol showed less than 10% difference in the results between the proposed automated method and a human expertise, 75% of which can be very easily corrected. Highlights ► A machine learning algorithm was developed to determine the chemical nature of microplastics. ► This method allows a fast and reliable automated identification even when several thousand of FTIR spectra have to be studied.► This method is the first part of a software dedicated to the study of microplastics: POSEIDON.

Since the 1950s plastic production has exponentially increased. Different expeditions have sampled microplastics in all the world’s oceans in order to better know this pollution. However, a large amount of plastic particles is collected during all campaigns, such as during the Tara Mediterranean Sea campaign. And, methods developed to analyse them are time-consuming. So, how to work to chemically characterize samples from large libraries?

The emergence of ocean acidification as a significant threat to calcifying organisms in marine ecosystems creates a pressing need to understand the physiological and molecular mechanisms by which calcification is affected by environmental parameters. We report here, for the first time, changes in gene expression induced by variations in pH/pCO(2) in the widespread and abundant coccolithophore Emiliania huxleyi. Batch cultures were subjected to increased partial pressure of CO2 (pCO(2); i.e. decreased pH), and the changes in expression of four functional gene classes directly or indirectly related to calcification were investigated. Increased pCO(2) did not affect the calcification rate and only carbonic anhydrase transcripts exhibited a significant down-regulation. Our observation that elevated pCO(2) induces only limited changes in the transcription of several transporters of calcium and bicarbonate gives new significant elements to understand cellular mechanisms underlying the early response of E. huxleyi to CO2-driven ocean acidification.

The study site located in the northwestern Mediterranean Sea was visited nine times in 2005–2006 to collect water samples from the epi- (5 m), meso- (200, 600 m), and bathypelagic (1000, 2000 m) zone. Total abundance of prokaryotes and viruses was determined by flow cytometry (FCM). Prokaryotic abundance in the epi-, meso-, and bathypelagic varied between 0.9 and 15.9×10 5 , 0.6 and 2.1×10 5 , and 0.3 and 1.3×10 5 ml −1 , respectively. Variation of viral abundance in the epi-, meso-, and bathypelagic was between 1.2 and 57.5×10 6 , 0.5 and 3.5×10 6 , and 0.4 and 1.3×10 6 ml −1 , respectively. The fraction of low (LNA) and high (HNA) nucleic acid prokaryotes averaged 42.9% and 57.1% throughout the water column and did not differ between depth layers. Throughout the water column the fraction of low, medium, and high fluorescent viruses (Vir-LF, Vir-MF, Vir-HF) averaged 66.3%, 30.2%, and 3.5%. Vir-LF and Vir-MF did not differ between depth layers; however, Vir-HF showed a preference for surface waters. The fraction of LNA cells decreased in the epi- and increased in the bathypelagic with decreasing stratification. The fraction of Vir-LF viruses increased in the epipelagic and decreased in the bathypelagic with increasing prokaryotic abundance. Also, the relationship between viral abundance and the bacterial community was different in surface and deep waters. The data suggest that different mechanisms of interaction between viruses and their prokaryotic hosts prevail at the surface and in deep waters.

Viruses on organic aggregates such as transparent exopolymeric particles (TEP) are not well investigated. The number of TEP-attached viruses was assessed along trophic gradients in the southwestern lagoon of New Caledonia by determining the fraction of viruses removed after magnetic isolation of TEP. In order to isolate TEP magnetically, TEP were formed in the presence of magnetic beads from submicrometer precursors collected along the trophic gradients. The mixed aggregates of TEP-beads-viruses were removed from solution with a magnetic field. The percentage of viruses associated with newly formed TEP averaged 8% (range, 3 to 13%) for most of the stations but was higher (ca. 30%) in one bay characterized by the low renewal rate of its water mass. The number of viruses ( N ) attached to TEP varied as a function of TEP size ( d [in micrometers]) according to the formulas N = 100 d 1.60 and N = 230 d 1.75 , respectively, for TEP occurring in water masses with short (i.e., 40 days) residence times. These two relationships imply that viral abundance decreases with TEP size, and they indicate that water residence time influences viral density and virus-bacterium interactions within aggregates. Our data suggest that the fraction of viruses attached to TEP is highest in areas characterized by a low renewal rate of the water mass and can constitute at times a significant fraction of total virus abundance. Due to the small distance between viruses and hosts on TEP, these particles may be hot spots for viral infection.

Reproducible and nondegradative preparation of sulfated molecules exhibiting diverse biological properties requires caring out sulfation reactions under ready controlled mild conditions. Although, to date, sulfur trioxide is the most used sulfation agent, its highly acid character has brought its use in association with different nitrogen bases as sulfur trioxide−nitrogen base complexes and/or by introduction of basic solvents as pyridine. We have applied sulfur trioxide and other sulfation agents including protocols for the synthesis of biologically active sulfated polysaccharides and demonstrated that these agents provoke cleavage of glycosidic bonds and other acid labile functions as amides, esters and even ethers. These facts prompted us to develop new reaction conditions for a general and nondestructive sulfation protocol. Our approach consists of the introduction of 2-methyl-2-butene as an acid scavenger of neutral character. Application of the method leads to an efficient, reproducible, and control...

The production of transparent exopolymeric particles (TEP) by the coccolithophores, Emiliania huxleyi, Calcidiscus leptoporus and Syracosphaera pulchra was investigated in batch cultures. The abundance, size spectra and carbon content of TEP were examined during the exponential growth phase of both haploid and diploid life stages grown under ambient (400 atm) and elevated (760 atm) CO2 partial pressure (pCO(2)) conditions. Results showed species- and life stage-specific differences in TEP production rate (day(1)) derived from abundance and carbon content of TEP. At 400 atm, TEP production rate was the highest in the diploid stage of S. pulchra and E. huxleyi, while TEP carbon content per cell was the highest in the diploid stage of C. leptoporus. At 760 atm, TEP production rate increased in almost all species and was closely related to the cell growth rates (except in the diploid stage of C. leptoporus), while the slope values of the regression lines between TEP size distribution and concentration decreased. This means that the contribution of smaller size TEP was relatively more important than larger TEP in the high pCO(2) treatment. Elevated pCO(2) is potentially able to alter TEP size distribution. TEP-C content cell(1) generally decreased with increasing pCO(2). TEP-C accounted for 124 of the cell particulate organic carbon production and was inversely related to increasing pCO(2). TEP production by C. leptoporus and S. pulchra has not previously been documented. The amount of organic carbon released as TEP by these coccolithophores is comparable to and may even exceed TEP production by some diatoms.

SUMMARYOcean acidification is predicted to have significant effects on benthic calcifying invertebrates, in particular on their early developmental stages. Echinoderm larvae could be particularly vulnerable to decreased pH, with major consequences for adult populations. The objective of this study was to understand how ocean acidification would affect the initial life stages of the sea urchin Paracentrotus lividus, a common species that is widely distributed in the Mediterranean Sea and the NE Atlantic. The effects of decreased pH (elevated PCO2) were investigated through physiological and molecular analyses on both embryonic and larval stages. Eggs and larvae were reared in Mediterranean seawater at six pH levels, i.e. pHT 8.1, 7.9, 7.7, 7.5, 7.25 and 7.0. Fertilization success, survival, growth and calcification rates were monitored over a 3 day period. The expression of genes coding for key proteins involved in development and biomineralization was also monitored. Paracentrotus lividus appears to be extremely resistant to low pH, with no effect on fertilization success or larval survival. Larval growth was slowed when exposed to low pH but with no direct impact on relative larval morphology or calcification down to pHT 7.25. Consequently, at a given time, larvae exposed to low pH were present at a normal but delayed larval stage. More surprisingly, candidate genes involved in development and biomineralization were upregulated by factors of up to 26 at low pH. Our results revealed plasticity at the gene expression level that allows a normal, but delayed, development under low pH conditions.

Bacterial abundance and community composition were investigated along trophic gra- dients in the barrier reef lagoon of Noumea, New Caledonia. Bacterial abundance and the percent- age of high nucleic acid (%HNA) bacteria (a potential indicator for bacterial production) increased from offshore waters towards the head of the bays. 16S rRNA gene PCR and denaturing gradient gel electrophoresis (DGGE) were used as genetic fingerprints for assessing differences in bacterial com- munity composition. Sequences of DGGE bands were assigned to (1) the genera Rugeria and Roseobacter (Rhodobacteriaceae), (2) the SAR11 cluster, (3) other Alphaproteobacteria, and (4) the genus Alteromonas. Removal of the operationally defined attached bacteria by prefiltration did not affect community profiles in offshore waters but had a strong influence in the bays, probably due to the much higher particle load and thus, attached bacteria in the bays. For the free-living community, the number of bands decreased linearly with increasing water residence time, chlorophyll a concen- tration, and viral abundance. Specific bands were found for offshore waters and the 2 investigated semi-enclosed bays, whereas the lagoon showed no specific bands. A similarity analysis showed spe- cific clusters for offshore water, the lagoon, and the bays. A principle component analysis together with cluster and correlation analysis indicated that water residence time, viruses, and a complex top- down cascading effect of ciliate grazers on flagellates influenced community composition. Also, data from fingerprints of the total and free-living communities suggest that the free-living and the attached community are controlled by different mechanisms.

The expression of genes of biogeochemical interest in calcifying and noncalcifying life stages of the coccolithophore Emiliania huxleyi was investigated. Transcripts potentially involved in calcification were tested through a light-dark cycle. These transcripts were more abundant in calcifying cells and were upregulated in the light. Their application as potential candidates for in situ biogeochemical proxies is also suggested.

International audience; Dilution experiments were carried out to investigate the community composition and the metabolic response of seawater and freshwater bacteria to cross-transplantation, and the effects of nor.-indigenous bacterial hosts on viral dynamics. Changes in viral and bacterial abundance and production, as Well as bacterial respiration, carbon demand and diversity were regularly monitored over a 6 d period. Bacterial production in the transplanted seawater (SB-t) and freshwater (FB-t) bacteria treatments was stimulated up to 256 and 221 %, respectively, compared to controls. The stimulation of bacterial production and carbon demand was accompanied by a decrease in bacterial richness. Net viral production was stimulated by 81% in SB-t and repressed by 75% in FB-t. Transplantation increased the virus-induced mortality of marine bacteria, but decreased it for freshwater bacteria. These results suggest that (1) marine bacteria can readily oxidize freshwater dissolved organic matter, and (2) freshwater viruses might be able to infect marine hosts, thus highlighting their potential role in fueling bacterial growth under resource stress or nutrient-depleted conditions.

Nonhealing wounds remain a major health problem whose treatment is challenging and costly. Treatments based on cells or growth factors are still not very effective. We developed an entirely novel strategy consisting in treatment of the wound-tissue matrix with biopolymers engineered to mimic heparan sulfates called OTR4120. This compound was dextran polymer with sulfated and carboxymethyl groupments. After binding to matrix proteins, the heparan-sulfate-mimicking polymer protects the microenvironment, maintaining the normal production of signals and growth factors needed for healing to occur. Here, we show that a specific biopolymer accelerates ulcer closure and improves re-epithelialization and dermal-matrix-component remodeling. OTR4120 treatment was associated with faster maturation of epidermal structures, most notably regarding the number of epithelial-cell layers, and with an appearance that more closely resembled normal skin. Treatment had also a main effect on collagen I and III expression. Necrotic skin ulcers induced in mice with doxorubicin recovered normal collagen levels and organization, with no evidence of fibrosis. Thus, appropriate polymer-based matrix therapy is a valid and simple alternative to regenerative medicine.

Burn-related skin fibrosis leads to loss of tissue function and hypertrophic scar formation with damaging consequences for the patient. There is therefore a great need for an efficient agent to treat burned skin. We report that ReGeneraTing Agent (RGTA) reduces burn-induced skin alteration. The tissue-regenerating effect of RGTA OTR4120 was evaluated after 1-6 days and after 10 months in a rat skin burn model. This effect was also examined in vitro using fibroblasts isolated from control and 6-day-old burned skins. We measured production of dermal collagen I, III, and V and activities of metalloproteinases 2 and 9 (MMP-2 and MMP-9). Ratio of collagen III over collagen I production increased 6 days after the burn, because of a decrease in collagen I production. After 10 months, ratio of collagen III over collagen I in burn sites was still increased compared with control skin, because of an increase in collagen III production. Both abnormalities were corrected by OTR4120. OTR4120 increased pro- and active MMP-2 and MMP-9, compared with healthy and burned controls and therefore accelerated remodeling. Similar data were obtained with cultured fibroblasts from healthy and burned skins. OTR4120 enhanced healing in short- and long-term after burns, reducing the formation of fibrotic tissue, and then represents a potential agent to improve burned skin healing. (c) 2006 Wiley Periodicals, Inc. J Biomed Mater Res, 2006.

Heparin affin regulatory peptide (HARP) is an heparin-binding growth factor, highly expressed in several primary human tumors and considered as a rate-limiting angiogenic factor in tumor growth, invasion, and metastasis. Implication of this protein in carcinogenesis is linked to its mitogenic, angiogenic, and transforming activities. Recently, we have demonstrated that the C-terminal residues 111-136 of HARP are required for its mitogenic and transforming activities (Bernard-Pierrot, I., Delbe, J., Caruelle, D., Barritault, D., Courty, J., and Milhiet, P. E. (2001) J. Biol. Chem. 276, 12228-12234). In this paper, HARP deleted of its last 26 amino acids was shown to act as a dominant negative effector for its mitogenic, angiogenic, transforming, and tumor-formation activities by heterodimerizing with the wild type protein. Similarly, the synthetic corresponding peptide P111-136 displayed in vitro inhibition of wild type HARP activities, but in this case, the inhibition was mainly explained by the competition of the peptide with HARP for the binding to the extracellular domain of the high affinity ALK receptor.

Heparin-affin regulatory peptide (HARP) and Midkine (MK) belong to a family of growth/differentiation factors that have a high affinity for heparin. The involvement of these molecules in various proliferative diseases prompted us to develop an assay for measuring the concentrations of these factors in biological fluids and culture media. This report describes an immunoassay that uses only commercially available materials, based on the high affinity of certain molecules for heparin. It consists of adsorbing heparin-BSA covalent complexes to microtiter plate wells and to quantify the heparin bound HARP or MK by using appropriate antibody. The method is specific and measures concentrations ranging from 40-1200 pg/mL HARP and from 25-1200 pg/mL MK and various parameters are investigated. The within-assay coefficient of variation was less than 5% for both assays. The method was checked by measuring the concentrations of these growth factors in the sera of healthy humans and in patients with cancer. As previously reported, we confirmed that the serum concentrations of MK are higher in patients with tumours (n = 139) than in controls (n = 19). The synthesis of HARP and MK by various cells in culture was also analysed.

International audience; In this study, the accuracy and the precision corresponding to Li isotopic measurements of low level samples such as marine and coastal carbonates are estimated. To this end, a total of fifty‐four analyses of a Li‐pure reference material (Li7‐N) at concentrations ranging from 1 to 6 ng ml−1 were first performed. The average δ7Li values obtained for solutions with and without chemical purification were 30.3 ± 0.4‰ (2s, n = 19) and 30.2 ± 0.4‰ (2s, n = 36), respectively. These results show that the chosen Li chemical extraction and purification procedure did not induce any significant isotope bias. Two available carbonate reference materials (JCt‐1 and JCp‐1) were analysed, yielding mean δ7Li values of 18.0 ± 0.27‰ (2s, n = 6) and 18.8 ± 1.8‰ (2s, n = 9), respectively. Small powder aliquots (< 15 mg) of JCp‐1 displayed significant isotope heterogeneity and we therefore advise favouring JCt‐1 for interlaboratory comparisons. The second part of this study concerns the determination of δ7Li value for biogenic carbonate samples. We performed a total of twenty‐nine analyses of seven different tropical coral species grown under controlled and similar conditions (24.0 ± 0.1 °C). Our sample treatment prior to Li extraction involved removal of organic matter before complete dissolution in diluted HCl. Our results show (a) a constant δ7Li within each skeleton and between the different species (δ7Li = 17.3 ± 0.7‰), and (b) a Li isotope fractionation of −2‰ compared with inorganic aragonite grown under similar conditions. Comparison with literature data suggests a significant difference between samples living in aquaria and those grown in natural conditions. Finally, we investigate ancient (fossil) carbonate material and foraminifera extracted from marine sedimentary records. Different leaching procedures were tested using various HCl molarities. Results indicate that carbonate preferential dissolution must be carried out at an acid molarity < 0.18 mol l−1. Possible contamination from silicate minerals can be verified using the Al/Ca ratio, but the threshold value strongly depends on the carbonate δ7Li value. When the silicate/carbonate ratio is high in the sediment sample (typically > 2), contamination from silicates cannot be avoided, even at low HCl molarity (⪡ 0.1 mol l−1). Finally, bulk carbonate and foraminifera extracted from the same core sample exhibited significant discrepancies: δ7Li values of foraminifera were more reproducible but were significantly lower. They were also associated with lower Sr/Ca and higher Mn/Ca ratios, suggesting a higher sensitivity to diagenesis, although specific vital effects cannot be fully ruled out.