Assessment of Lipid Quality in Commercial Omega-3 Supplements Sold in the French Market.

Supplementation of omega-3 fatty acids is considered a valuable strategy to supply the low intake of these fatty acids. Thus, the safety of the supplements is an important milestone. Because of that, we analyzed 20 unflavored supplements sold in the French market for fatty acid and triglyceride composition, for EPA and DHA, and for tocol content, as well as for oxidative status. This study found that only 2.5% of the supplements did not meet their label claims for omega-3 content. TAG analysis showed high variability among the triglyceride distribution, and the same trend was also noticed for the tocol content; in fact, a high variability of the distribution of the six tocols (four tocopherols and two tocotrienols) was found among the samples. Of the tested products, all of them complied with peroxide value, p-anisidine value, and Totox values established by the Global Organization for EPA and DHA Omega-3s (GOED) and were not oxidized.

Independence of a Marine Unicellular Diazotroph to the Presence of NO3.

Marine nitrogen (N2) fixation was historically considered to be absent or reduced in nitrate (NO3-) rich environments. This is commonly attributed to the lower energetic cost of NO3- uptake compared to diazotrophy in oxic environments. This paradigm often contributes to making inferences about diazotroph distribution and activity in the ocean, and is also often used in biogeochemical ocean models. To assess the general validity of this paradigm beyond the traditionally used model organism Trichodesmium spp., we grew cultures of the unicellular cyanobacterium Crocosphaera watsonii WH8501 long term in medium containing replete concentrations of NO3-. NO3- uptake was measured in comparison to N2 fixation to assess the cultures' nitrogen source preferences. We further measured culture growth rate, cell stoichiometry, and carbon fixation rate to determine if the presence of NO3- had any effect on cell metabolism. We found that uptake of NO3- by this strain of Crocosphaera was minimal in comparison to other N sources (~2-4% of total uptake). Furthermore, availability of NO3- did not statistically alter N2 fixation rate nor any aspect of cell physiology or metabolism measured (cellular growth rate, cell stoichiometry, cell size, nitrogen fixation rate, nitrogenase activity) in comparison to a NO3- free control culture. These results demonstrate the capability of a marine diazotroph to fix nitrogen and grow independently of NO3-. This lack of sensitivity of diazotrophy to NO3- suggests that assumptions often made about, and model formulations of, N2 fixation should be reconsidered.

In-source fragmentation of very labile peptides in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

Synthetic acidic proline-rich peptides devoid of basic chemical groups were studied by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF). Their ion mass spectra recorded in reflector positive ion mode have shown unusual features, i.e., absence or very weak presence of protonated peptide together with a major peak associated with fragmentation at a site that corresponds to the amide bond N-terminal to the first proline of the XPP motif. In contrast, arginine-containing analogues were stable in MALDI-TOF, whereas peptides sharing a free N-terminal amino group were moderately subject to the same fragmentation. Effects of extraction delay time suggest that this process takes place very early (nanoseconds) at the beginning of the plume expansion. The effect of the nature of the matrix on the survival yield indicates a better correlation with the initial axial velocity than with the matrix proton affinity. All the data show some strong differences with the classical in-source decay (ISD). Our results suggest the role of the available protons in the close neighborhood of the peptide during the crystallization process and the prompt fragmentation induced by collisions in the first step of ablation. Undoubtedly, our study highlights that the MALDI-TOF analysis of peptides containing proline and no basic group should be carried out with extreme caution.

Isolation, characterization and molecular cloning of new temporins from the skin of the North African ranid Pelophylax saharica.

Temporins are small antimicrobial peptides isolated from North American and Eurasian ranid frogs that are particularly active against Gram-positive bacteria. To date, no temporins have been characterized from North African frog species. We isolated three novel members of the temporin family, named temporin-1Sa (FLSGIVGMLGKLF(amide)), -1Sb (FLPIVTNLLSGLL(amide)), and -1Sc (FLSHIAGFLSNLF(amide)), from the skin of the Sahara frog Pelophylax (Rana) saharica originating from Tunisia. These temporins were identified by a combined mass spectrometry/molecular cloning approach. Temporin-1Sa was found to be highly active against Gram-positive and Gram-negative bacteria, yeasts and fungi (MIC=2-30 microM). To our knowledge, this is the first 13-residue member of the temporin family with a net charge of +2 that shows such broad-spectrum activity with particularly high potency on the clinically relevant Gram-negative strains, Escherichia coli (MIC=10 microM) and Pseudomonas aeruginosa (MIC=31 microM). Moreover, temporin-1Sa displays significant antiparasitic activity (IC50 approximately 20 microM) against the promastigote and amastigote stages of Leishmania infantum.

Protein desolvation in UV matrix-assisted laser desorption/ionization (MALDI).

We describe experiments in MALDI-TOF and MALDI-TOF-TOF showing that the ejection of protein-matrix cluster ions and their partial decay in the source occur in MALDI. The use of radial beam deflection and small size detector in linear mode allows detection of ions with higher time-of-flight and kinetic energy deficit. MALDI-TOF-TOF experiments were carried out by selecting chemical noise ions at m/z higher than that of a free peptide ion. Whatever the selected m/z (up to m/z 300) the molecular peptide ion appeared as the main fragment. The production of protein-matrix clusters and their partial decay in the source was found to increase with the size of the protein (MW from 1000 to 150,000 u), although it decreases with increasing charge state. These effects were observed for different matrices (HCCA and SA) and in a large laser fluence range. Experimental results and calculation highlight that a continuous decay of protein-matrix cluster ions occurs in the source. This decay-desolvation process can account for the high-mass tailing and peak shifting as well as the strong noise/background in the mass spectra of proteins.

Uptake of dissolved inorganic and organic nitrogen by the benthic toxic dinoflagellate Ostreopsis cf. ovata.

Environmental factors that shape dynamics of benthic toxic blooms are largely unknown. In particular, for the toxic dinoflagellate Ostreopsis cf. ovata, the importance of the availability of nutrients and the contribution of the inorganic and organic pools to growth need to be quantified in marine coastal environments. The present study aimed at characterizing N-uptake of dissolved inorganic and organic sources by O. cf. ovata cells, using the 15N-labelling technique. Experiments were conducted taking into account potential interactions between nutrient uptake systems as well as variations with the diel cycle. Uptake abilities of O. cf. ovata were parameterized for ammonium (NH4+), nitrate (NO3-) and N-urea, from the estimation of kinetic and inhibition parameters. In the range of 0 to 10µmolNL-1, kinetic curves showed a clear preference pattern following the ranking NH4+>NO3->N-urea, where the preferential uptake of NH4+ relative to NO3- was accentuated by an inhibitory effect of NH4+ concentration on NO3- uptake capabilities. Conversely, under high nutrient concentrations, the preference for NH4+ relative to NO3- was largely reduced, probably because of the existence of a low-affinity high capacity inducible NO3- uptake system. Ability to take up nutrients in darkness could not be defined as a competitive advantage for O. cf. ovata. Species competitiveness can also be defined from nutrient uptake kinetic parameters. A strong affinity for NH4+ was observed for O. cf. ovata cells that may partly explain the success of this toxic species during the summer season in the Bay of Villefranche-sur-mer (France).

MALDI TOF-TOF characterization of a light stabilizer polymer contaminant from polypropylene or polyethylene plastic test tubes.

Disposable plasticware such as plastic test tubes are routinely used in all proteomics laboratories. Additives in polymers are used to protect them against oxygen or ultraviolet (UV) light degradation. Hindered amine light stabilizers (HALSs) are of utmost importance in modern polyolefin (polypropylene, polyethylene) stabilization. In this article, we demonstrate that the manufacturing polymeric agent: poly-(N-beta-hydroxyethyl-2,2,6,6-tetramethyl-4-hydroxy-piperidinyl succinate), known as Tinuvin-622 or Lowilite 62, from the HALS family, leaches from laboratory polypropylene or polyethylene plastic test tubes into the standard solvents for sample preparation. The analysis of these polluted samples by matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) mass spectrometry, in the positive mode, shows highly contaminated mass spectra, due to the high sensitivity of this technique. These contaminants have mass range and mass defect similar to those of peptides arising from the digestion of a protein in a conventional proteomics study. Therefore, they can be really harmful for proteomics studies, leading to misattributions, preventing any protein identification. In this article, an MS and MS/MS fingerprint of this pollutant is given and some pieces of advice to avoid it are proposed.

Characterization of ecdysteroids in Drosophila melanogaster by enzyme immunoassay and nano-liquid chromatography-tandem mass spectrometry.

Ecdysteroids are polyhydroxylated steroids that function as molting hormones in insects. 20-Hydroxyecdysone (a 27C-ecdysteroid) is classically considered as the major steroid hormone of Drosophilamelanogaster, but this insect also contains 28C-ecdysteroids. This arises from both the use of several dietary sterols as precursors for the synthesis of its steroid hormones, and its inability to dealkylate the 28C-phytosterols to produce cholesterol. The nature of Drosophila ecdysteroids has been re-investigated using both high-performance liquid chromatography coupled to enzyme immunoassay and a particularly sensitive nano-liquid chromatography-mass spectrometry methodology, while taking advantage of recently available ecdysteroid standards isolated from plants. In vitro incubations of the larval steroidogenic organ, the ring-gland, reveals the synthesis of ecdysone, 20-deoxy-makisterone A and a third less polar compound identified as the 24-epimer of the latter, while wandering larvae contain the three corresponding 20-hydroxylated ecdysteroids. This pattern results from the simultaneous use of higher plant sterols (from maize) and fungal sterols (from yeast). The physiological relevance of all these ecdysteroids, which display different affinities to the ecdysteroid receptors, is still a matter of debate.