The antioxidant activity and transcellular pathway of Asp‐Leu‐Glu‐Glu in a Caco‑2 cell monolayer

Asp‐Leu‐Glu‐Glu (DLEE) is one of the antioxidant peptides purified from Chinese dry‐cured Xuanwei ham in our previous study. In the current work, the stability in a simulated digestion system, the transportation pathway and the antioxidant ability of DLEE were further investigated in a Caco‐2 cell monolayer. In the simulated gastrointestinal digestion system, no oligopeptides were generated. In the transport trial, the inhibitors cytochalasin D increased the transport of DLEE across the Caco‐2 cell monolayer, with P_app values of 3.22 × 10^?6 cm s^?1. A decreased expression occludin was observed with the DLEE incubation in the cell monolayer, and the antioxidant activity showed to be increased gradually in basolateral side. This study indicates that the absorption of DLEE could mainly occur via paracellular transport and provides information about its antioxidant activity after being absorbed across a cell monolayer.     

S‐Allyl cysteine,S‐ethyl cysteine and S‐propyl cysteine alleviate oxidative stress‐induced damage within PC12 cells

BACKGROUND: The PC12 cell line is a suitable model for the investigation of neurodegenerative diseases. In this study, PC12 cells were used to examine in vitro antioxidative and antiapoptotic protection by S‐allyl cysteine (SAC), S‐ethyl cysteine (SEC) and S‐propyl cysteine (SPC). PC12 cells were treated with these agents at 5 and 10 µmol L^?1 before exposure to hydrogen peroxide (H₂O₂). RESULTS: H₂O₂ treatment significantly decreased mitochondrial membrane potential (MMP) and cell viability and increased lactate dehydrogenase (LDH) release and DNA fragmentation (P < 0.05). The pre‐treatments with SAC, SEC and SPC significantly and concentration‐dependently elevated cell viability and MMP and lowered LDH release and DNA fragmentation (P < 0.05). H₂O₂ treatment also significantly increased levels of malondialdehyde (MDA), reactive oxygen species (ROS) and oxidised glutathione (GSSG) and decreased glutathione (GSH) content (P < 0.05). The pre‐treatments with SAC, SEC and SPC significantly decreased subsequent H₂O₂‐induced formation of MDA, ROS and GSSG (P < 0.05) and also alleviated H₂O₂‐induced GSH depletion (P < 0.05). Finally, H₂O₂ treatment significantly decreased Na⁺‐K⁺‐ATPase activity and elevated caspase‐3 activity (P < 0.05). The pre‐treatments with SAC, SEC and SPC significantly attenuated H₂O₂‐induced Na⁺‐K⁺‐ATPase activity reduction and caspase‐3 activity elevation (P < 0.05). CONCLUSION: The results obtained support that the three cysteine‐containing compounds studied are potent neuroprotective agents. Copyright © 2008 Society of Chemical Industry     

Low‐temperature wine‐making using yeast immobilized on pear pieces

A biocatalyst was prepared by the immobilization of Saccharomyces cerevisiae AXAZ‐1 yeast cells on pear pieces and tested for grape must fermentation in both batch and continuous conditions. The immobilized yeast cells were stable and active even at low temperatures (<10 °C). Wine production under batch fermentation at 8 °C was completed within 15 days while at 3 °C it took 36 days. In continuous fermentation, the bioreactor was operated for 33 days, then stored for 12 days at 10 °C, and re‐run for another 15 days without any diminution of the ethanol productivity. Total acidity of the produced wines remained within the ranges usually observed in dry wines, while volatile acidity was found in rather increased levels. The concentrations of higher alcohols (1‐propanol, isobutyl alcohol and amyl alcohols) were relatively low, while ethyl acetate was detected at up to 118 mg l^?1, contributing to the fruity aroma of the wines produced. Preliminary sensory evaluations carried out in the laboratory indicated the fine quality of the produced wines. Copyright © 2004 Society of Chemical Industry     

Determination of S‐methyl‐L‐methionine (SMM) from Brassicaceae Family Vegetables and Characterization of the Intestinal Transport of SMM by Caco‐2 Cells

The objectives of the current study were to determine S‐methyl‐L‐methionine (SMM) from various Brassicaceae family vegetables by using validated analytical method and to characterize the intestinal transport mechanism of SMM by the Caco‐2 cells. The SMM is well known to provide therapeutic activity in peptic ulcers. The amount of SMM from various Brassicaceae family vegetables ranged from 89.08 ± 1.68 μg/g to 535.98 ± 4.85 μg/g of dry weight by using validated ultra‐performance liquid chromatography‐electrospray ionization‐mass spectrometry method. For elucidating intestinal transport mechanism, the cells were incubated with or without transport inhibitors, energy source, or a metabolic inhibitor. Phloridzin and verapamil as inhibitors of sodium glucose transport protein (SGLT1) and P‐glycoprotein, respectively, were not responsible for cellular uptake of SMM. Glucose and sodium azide were not affected by the cellular accumulation of SMM. The efflux ratio of SMM was 0.26, implying that it is not effluxed through Caco‐2 cells. The apparent coefficient permeability (P _app) of SMM was 4.69 × 10^?5 cm/s, indicating that it will show good oral absorption in in vivo .     

Degradation of N‐(1‐Deoxy‐d‐xylulos‐1‐yl)glycine and N‐(1‐Deoxy‐d‐xylulos‐1‐yl)proline using thermal treatment

The formation and degradation of N‐(1‐Deoxy‐d ‐xylulos‐1‐yl)glycine and N‐(1‐Deoxy‐d ‐xylulos‐1‐yl)proline, derived from the secondary amine Maillard reaction in xylose‐amino acid model solutions, were detailed in this study. The identification and quantitative analysis of N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)glycine and N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)proline were carried out using high‐performance anion‐exchange chromatography and high‐performance liquid chromatography. The formation of intermediate and advanced products derived from N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)glycine and N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)proline was also tested using an UV‐Vis spectrophotometer to gain a better comparing of the degradation process of the two important Maillard reaction products using thermal treatment. Results showed that the degradation of N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)glycine was more significant than N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)proline. Moreover, xylose was tested in the degradation products of both N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)glycine and N‐(1‐Deoxy‐d‐ xylulos‐1‐yl)proline, which indicated that the degradation of N‐substituted 1‐amino‐1‐deoxyketoses was a reversible reaction to form reducing sugar.     

Formation of protein adducts of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine in cooked foods

Heterocyclic amines (HCAs) are mutagenic and carcinogenic compounds found in cooked meat and fish. Although HCAs are known to form adducts with protein after metabolic activation, adduct formation during cooking has not been elucidated. In this study, we showed that 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) is released from high molecular weight compounds by acid or enzymatic hydrolysis of cooked foods. Formation of free and protein adduct forms of PhIP was dependent on cooking temperature and time, and PhIP–protein adducts were estimated to form after formation of free PhIP. We also demonstrated that PhIP–protein adduct is formed by heating of PhIP and albumin as a model protein. A new adduct peak including [M+H]⁺ (m/z=225) of PhIP as a fragment ion was detected in the high molecular weight fraction of heat‐treated protein by LC–MS analysis. From model experiments by heating of PhIP and amino acids, the adduct was estimated to be produced by condensation of the amino group of PhIP and the carboxyl group of protein. PhIP–protein adducts were detected in several cooked meat and fish at ng/g food level as PhIP content. These results suggest that food‐borne protein adducts of HCAs may influence human HCA exposure and carcinogenic risk.