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Quantitative proteomic changes in the liver of adult males of Sheepshead minnow (Cyprinodon variegatus) upon exposure to ethinyl estradiol (EE2) were assessed to provide an advanced understanding of the metabolic pathways affected by estrogenic endocrine disruption in marine fish, and to identify potential novel molecular biomarkers for the environmental exposure to estrogens. From a total of 3188 identified protein groups (hereafter proteins), 463 showed a statistically significant difference in their abundance between EE2 treatment and solvent control samples. The most affected biological processes upon EE2 exposure were related to ribosomal biogenesis, protein synthesis and transport of nascent proteins to endoplasmic reticulum, and nuclear mRNA catabolism. Within the group of upregulated proteins, a subset of 14 proteins, involved in egg production (Vitellogenin, Zona Pellucida), peptidase activity (Cathepsine E, peptidase S1, Serine/threonine-protein kinase PRP4 homolog, Isoaspartyl peptidase and Whey acidic protein), and nucleic acid binding (Poly [ADP-ribose] polymerase 14) were significantly upregulated with fold-change values higher than 3. In contrast, Collagen alpha-2, involved in the process of response to steroid hormones, among others, was significantly downregulated (fold change = 0.2). This pattern of alterations in the liver proteome of adult males of C. variegatus can be used to identify promising novel biomarkers for the characterization of exposure of marine fish to estrogens. The Whey acidic protein-like showed the highest upregulation in EE2-exposed individuals (21-fold over controls), suggesting the utility of abundance levels of this protein in male liver as a novel biomarker of xenoestrogen exposure.
This study investigated the effects of domestic cooking process on the variations of soybean isoflavones, aiming at understanding the conversion of β-glucosides and aglycones isoflavones during the process and the relation with antioxidant activity. It was found that β-glucosides isoflavones was significantly increased from 223.01 (raw) to 727.29 mg/g (frying at 160 °C for 2 min), but boiling showed only a slight increase to 258.14 mg/g. The process for the mixed cooking of soybeans with vegetables was also evaluated, which is quite common in home cuisine. The results showed all bioactive ingredients were aggressively destroyed by over processing, but interestingly, green pepper and kelp exhibited isoflavones generation potentials for soybean. In addition, cooking from 60 to 160 °C for 2 or 5 min, showed a significantly decrease on FRAP. However, in the case of fried soybeans which treated at 120 °C or 160 °C, when extending the heating time to 5 min, their FRAP activity got a significant increase. The present study may provide a practical guidance for healthy soybean cooking, by using frying around 120 °C for 5 min and mixed with some vegetables such as green peppers or kelp. Keywords: Soybeans, Isoflavones, Cooking processing, Antioxidant activity
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