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The effect of natural antioxidants was tested by oxidative stability index (OSI) in refined soybean oil (RSO) using Rancimat method. Phenolic compounds of Thompson grape pomace were extracted with 95:5 (v : v) ethanol : water. The antioxidant activity of grape pomace extract (GPE) obtained was compared with those of rosemary extract (RE), and tocopherols mix (TM) known as covi‐ox T‐70. The synergistic effect of citric acid was also evaluated. Results showed a total phenols content of 17.39 mg (+)‐catechin equivalents per g of GPE. Thompson GPE at 0.3 and 0.5% (w/w) exhibited greater antioxidant activity than TM. RSO containing GPE or RE at 0.5% (w/w) showed an OSI higher than 48 h at 110C. Citric acid did not show synergistic effect with GPE. However, a synergistic effect for TM at 0.02% (w/w) with citric acid at 1.0% (w/w) was observed. Citric acid at 2.0% (w/w) with RE at all concentrations tested also displayed synergism. PRACTICAL APPLICATIONS Synthetic antioxidants are widely used in the food industry to delay fat oxidation; however, their utilization has been questioned due to toxicology concerns. Researchers have recognized the need to identify new natural antioxidants to use as additives. In this study, we measured total phenolics content of Thompson grape pomace extract (GPE) and found that the antioxidant activity of this extract was stronger than tocopherols from soybean oil. We propose that GPE can have applications as a food antioxidant.

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Hypoxia is a frequent source of stress in the estuarine habitat of the white shrimp Litopenaeus vannamei. During hypoxia, L. vannamei gill cells rely more heavily on anaerobic glycolysis to obtain ATP. This is mediated by transcriptional up-regulation of glycolytic enzymes including glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The hypoxia inducible factor 1 (HIF-1) is an important transcriptional activator of several glycolytic enzymes during hypoxia in diverse animals, including crustaceans. In this work, we cloned and sequenced a fragment corresponding to the 5′ flank of the GAPDH gene and identified a putative HIF-1 binding site, as well as sites for other transcription factors involved in the hypoxia signaling pathway. To investigate the role of HIF-1 in GAPDH regulation, we simultaneously injected double-stranded RNA (dsRNA) into shrimp to silence HIF-1α and HIF-1β under normoxia, hypoxia, and hypoxia followed by reoxygenation, and then measured gill HIF-1α, HIF-1β expression, GAPDH expression and activity, and glucose and lactate concentrations at 0, 3, 24 and 48 h. During normoxia, HIF-1 silencing induced up-regulation of GAPDH transcripts and activity, suggesting that expression is down-regulated via HIF-1 under these conditions. In contrast, HIF-1 silencing during hypoxia abolished the increases in GAPDH expression and activity, glucose and lactate concentrations. Finally, HIF-1 silencing during hypoxia-reoxygenation prevented the increase in GAPDH expression, however, those changes were not reflected in GAPDH activity and lactate accumulation. Altogether, these results indicate that GAPDH and glycolysis are transcriptionally regulated by HIF-1 in gills of white shrimp.

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