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  • Publication
    Accès libre
    Sensitive and selective quantification of free and total malondialdehyde in plasma using UHPLC-HRMS
    Quantification of malondialdehyde (MDA) as a marker of lipid peroxidation is relevant for many research fields. We describe a new sensitive and selective method to measure free and total plasmatic MDA, using derivatization with 2,4-dinitrophenylhydrazine (DNPH) and ultrahigh performance liquid chromatography-high resolution mass spectrometry (UHPLC-HRMS). Free and total MDA were extracted from minute sample amounts (10 μL) using acidic precipitation and alkaline hydrolysis followed by acidic precipitation, respectively. Derivatization was completed within 10 min at room temperature, and the excess DNPH discarded by liquid-liquid extraction. Quantification was achieved by internal standardization using dideuterated MDA as internal standard. The method lowest limit of quantification (LLOQ) was 100 nM and linearity spanned > 3 orders of magnitude. Intra- and inter-day precisions for total MDA were 2.9% and 3.0%, respectively, and those for free MDA were 12.8% and 24.9%, respectively. Accuracy was 101% and 107% at low and high concentrations, respectively. In human plasma, free MDA levels were 120 nM (SD 36.26) and total MDA levels were 6.7 μM (SD 0.46). In addition, we show the applicability of this method to measure MDA plasma levels from a variety of animal species, making it invaluable to scientists in various fields.
  • Publication
    Métadonnées seulement
    Reproductive effort and oxidative stress: effects of offspring sex and number on the physiological state of a long-lived bird
    (2017-2-13)
    Merkling, Thomas
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    Blanchard, Pierrick
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    Chastel, Olivier
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    ;
    Vallat, Armelle
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    Hatch, Scott A.
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    Danchin, Etienne
    ;
    1. Individuals must trade-off between energetically costly activities to maximize their fitness. However, the underlying physiological mechanism remains elusive. Oxidative stress, the imbalance between reactive oxygen species production and antioxidant and/or repair activities, has been suggested to underlie life-history trade-offs: greater investment in reproduction supposedly generating higher oxidative damage, thus reducing life span. 2. While most studies used natural or experimental variation in offspring number to examine how reproduction affects oxidative stress, none studied the impact of offspring sex, although it could influence physiological costs and fitness, if the sexes differ in terms of energetic cost. 3. Here, we aim at further understanding how reproduction (in terms of offspring sex, experimentally manipulated and number, not manipulated) influences oxidative stress in a wild seabird, where sons are energetically costlier than daughters. We did so by conducting a chick fostering experiment (to disentangle foster and produced sex ratio) and using four oxidative stress markers plus baseline corticosterone. 4. First, the results suggest that individual physiological state before laying modulates upcoming reproductive effort. Individuals with higher pre-laying baseline corticosterone and lower antioxidant activity, estimated by their superoxide dismutase activity, subsequently invested more in reproduction, estimated by the cumulative number of days spent rearing chicks. Hence, it seems that only individuals that could afford to invest heavily in reproduction did so. 5. Then, we examined the effects of reproductive effort on individual physiological state at the end of the breeding season. Higher reproductive effort seemed to imply higher physiological costs. Oxidative stress, estimated by the ratio of oxidized over reduced glutathione, increased with more male-biased foster sex ratio among mothers but not among fathers, whereas baseline corticosterone did so in both sexes. Similarly, lipid oxidative damage to red blood cells increased with increasing cumulative number of days spent rearing chicks. 6. Our study provides the first evidence that brood sex ratio variation can affect oxidative balance, potentially in a sex-specific way, although more studies are needed to understand whether the observed physiological costs could lead to fitness costs. It also highlights the need to consider sex ratio in future studies investigating the role of oxidative stress in life-history trade-offs.