Subchronic toxicity and hepatocyte apoptosis of dietary olaquindox in common carp (Cyprinus carpio).

Olaquindox as one of the effective antimicrobial agents and growth-promoting feed additives, had been widely used in animal and fish production. However, few studies have been done to unveil its possible toxic effect and tissue injury on aquatic animal. In this study, the toxic effect and underlying mechanisms of olaquindox toxicity were investigated in common carp when feed with different doses of olaquindox for 90 days. The morbidity and mortality, pathological changes, hematology parameters, residue concentration in the tissues of common carp were assessed, hepatocyte apoptosis was detected through ultrastructural observation and flow cytometry methods. The results showed that the morbidity and mortality increased with the increasing dosages of dietary olaquindox, subchronic exposure to olaquindox caused remarkably pathological changes, including congestion and bleeding, intramuscular edema, vacuolar degeneration, degeneration and deformation in renal tubules architecture, respiratory epithelium fusion and intestinal epithelial microvilli disintegration. Besides, dietary olaquindox led to significant changes in blood biochemical parameters including red blood cell, hemoglobin, alanine aminotransferase and aspartate aminotransferase, an elevated residue concentration of olaquindox was detected in liver and kidney after exposure, hepatocyte apoptosis and necrosis were observed. Moreover, insulin-like growth factor I (IGF-I) mRNA level in liver was higher than normal level with the dose below 25 mg/kg olaquindox and was lower than normal level with the dose above 50 mg/kg. Our results demonstrated that dietary olaquindox may pose subchronic toxicity and residue in fish organs and provided scientific data for the safe application of olaquindox in fish.

α-Glucosidase inhibitors from brown rice bound phenolics extracts (BRBPE): Identification and mechanism.

Brown rice bound phenolics extracts (BRBPE) have been reported to possess α-glucosidase inhibitory effects, the specific enzyme inhibitors involved in this process were unknown. Here, α-glucosidase inhibitors in BRBPE were screened using bioaffinity ultrafiltration methods, and seven phenolic compounds - three monomers (p-coumaric acid, ferulic acid and methyl ferulate), three dimers (8-5', 5-5' and 8-O-4' diferulic acid) and a trimer (5-5'/8-O-4″ dehydrotriferulic acid) were identified as exact inhibitors, among which 5-5'/8-O-4″ dehydrotriferulic acid and 5-5'diferulic acid exhibited the best inhibitory activity. Enzyme kinetic analysis suggested that the inhibitory mechanism of these seven inhibitors including competitive, noncompetitive, uncompetitive and mixed manner. Molecular docking analysis revealed that the seven inhibitors bind with α-glucosidase mainly by hydrogen bonding interaction, hydrophobic force and ionic bond. Molecular dynamics simulation further explored the structure and molecular property of phenolic-glucosidase complex. This work provided a deep insight into brown rice bound phenolics acting as potent α-glucosidase inhibitors.

[Recombinant expression of black sesame polyphenol oxidase and its enzymatic properties].

To investigate the enzyme properties of the black sesame polyphenol oxidase (BsPPO), a synthesized Bsppo gene was cloned into the vector pMAL-c5x and expressed in E. coli. Subsequently, the MBP fusion label in the recombinant protein was removed by protease digestion after affinity purification. The synthesized Bsppo gene contained 1 752 bp which encodes 585 amino acids with a deduced molecular weight of 65.3 kDa. Transformation of the recombinant vector into E. coli BL21(DE3) resulted in soluble expression of the fusion protein MBP-BsPPO. The enzymatic properties of the recombinant BsPPO was investigated after MBP fusion tag excision followed by affinity purification. The results demonstrated that the optimal temperature and pH for BsPPO was 25°C and 4.0, respectively. BsPPO exhibited a good stability under low temperature and acidic environment. Low-intensity short-term light exposure increased the activity of BsPPO. Cu²âº could improve the activity of BsPPO while Zn²âº and Ca²âº showed the opposite effect. BsPPO could catalyze the oxidation of monophenols, diphenols, and triphenols, and exhibited good catalytic activity on l-tyrosine and vanillic acid. Moreover, BsPPO exhibited high catalytic activity on black sesame metabolites, including 2-methoxy cinnamic acid, indole-3-carboxylic acid and phloretin. These results may serve as a basis for further characterization of BsPPO.