Application of a solid-phase fluorescence immunoassay to determine oxytetracycline and tetracycline residues in tissue of olive flounder (Paralichthys olivaceus)

Parallux, a solid-phase fluorescence immunoassay (SPFIA) developed for antibotics residue detection in milk, was applied for analysis of fish tissue. The recommended therapeutic doses of oxytetracycline (OTC, 100 g/ton water, withdrawal period 30 days) and tetracycline (TC, 150 g/ton water, withdrawal period 30 days) were treated to a group of 35 olive flounders (Paralichthys olivaceus) using dipping administration. Muscle was sampled before and after drug treatment 1st, 2nd, 3rd, 5th, 7th, and 14th day. The concentration of oxytetracycline in muscle, determined by SPFIA, was compared with that of internal standard (100 ppb as oxytetracycline). The S/C ratio of sample inhibition value to cutoff inhibition value was employed as an index to determine the muscle residue in olive flounder. To investigate the recovery rate, and standard solutions were added to muscle samples to give final concentrations in muscle of 0.1 and 0.5 microg/ml. The recovery rates of all spiked samples were >89% of the spiked value. OTC and TC were detected in muscle of fishes treated until the 3rd day of withdrawal period. The present study showed that the SPFIA can be easily adopted in predicting tissue residues for OTC and TC in farmed fishes.     

Wheat bran oil and its fractions inhibit human colon cancer cell growth and intestinal tumorigenesis in Apc(min/+) mice

This study was designed to investigate the cancer preventive activities of wheat bran (WB) oil. We studied the colon cancer preventive effects of WB oil and its subfractions in the Apc(min/+) mouse model, a recognized mouse model for human colorectal cancer, and used human colon cancer cell lines (HCT-116 and HT-29) to identify possible active fractions in WB oil. Our results showed that the oil fraction of WB was more active than the water fraction against the growth of human colon cancer cell lines and that 2% WB oil significantly inhibited the overall tumorigenesis by 35.7% (p < 0.0001) in the Apc(min/+) mouse model. The WB oil was further fractioned into nonpolar lipids and phytochemicals and the phytochemical fraction was fractionated into phytosterols and phytosterol ferulates, 5-alk(en)ylresorcinols, and unidentified constituents by normal phase silica gel column chromatography. Results on cell culture showed that the phytochemical fraction had a higher inhibitory effect on HCT-116 human colon cancer cells than that of WB oil, whereas the nonpolar lipid fraction had less growth inhibitory effectiveness. However, neither fractions showed a stronger inhibition than WB oil in the Apc(min/+) mouse model. The current results demonstrate, for the first time, the intestinal cancer preventive activity of WB oil. The active ingredients, however, remain to be identified.     

Peroxynitrite scavenging activity of sinapic acid (3,5-dimethoxy-4-hydroxycinnamic acid) isolated from Brassica juncea

Peroxynitrite (ONOO(-)), formed from a reaction of superoxide and nitric oxide, is one of the most potent cytotoxic species that are known to oxidize cellular constituents including essential proteins, lipids, and DNA. In this study, the ability of sinapic acid (3,5-dimethoxy-4-hydroxycinnamic acid), isolated from Brassica juncea, to scavenge ONOO(-) was investigated. The data obtained show that sinapic acid can efficiently scavenge native ONOO(-) as well as ONOO(-) derived from the peroxynitrite donor 3-morpholinosydnonimine hydrochloride (SIN-1). Spectrophotometric analyses revealed that sinapic acid suppressed the formation of ONOO(-)-mediated tyrosine nitration through an electron donation mechanism. In further studies, sinapic acid also showed a significant ability of inhibiting nitration of bovine serum albumin and low-density lipoprotein (LDL) in a dose-dependent manner. Sinapic acid decreased the LDL peroxidation induced by SIN-1-derived ONOO(-). The present study suggests that sinapic acid has an efficient ONOO(-) scavenging ability, which may well be a potent ONOO(-) oxidant scavenger for the protection of the cellular defense activity against the ONOO(-)-involved diseases.     

Evaluation of antioxidant activity of Australian tea tree (Melaleuca alternifolia) oil and its components

Antioxidant activity of Australian tea tree (Melaleuca alternifolia) oil (TTO) was determined using two different assays. In the 2,2-diphenyl-1-picrylhydrazyl assay, 10 microL/mL crude TTO in methanol had approximately 80% free radical scavenging activity, and in the hexanal/hexanoic acid assay, 200 microL/mL crude TTO exhibited 60% inhibitory activity against the oxidation of hexanal to hexanoic acid over 30 days. These results were equivalent to the antioxidant activities of 30 mM butylated hydroxytoluene in both tests at the same experimental conditions. This indicated that the TTO could be a good alternative antioxidant. Inherent antioxidants, i.e., alpha-terpinene, alpha-terpinolene, and gamma-terpinene, in the crude TTO were separated and identified chromatographically using silica gel open chromatography, C(18)-high-pressure liquid chromatography, and gas chromatography-mass spectrometry. Their antioxidant activities decreased in the following order in both assays: alpha-terpinene > alpha-terpinolene > gamma-terpinene.     

Evaluation of citrinin occurrence and cytotoxicity in Monascus fermentation products

Monascus purpureus and its fermentation products have been used in food coloring and meat preservation in Asia for centuries and have also been recently used as dietary supplements because of their cholesterol-lowering ability. However, the presence of the mycotoxin citrinin (CTN), a secondary metabolite of Monascus species, in fermentation products is a potential threat to public health. In the present study, HPLC was used to analyze CTN levels in lipid and aqueous extracts of commercialized Monascus products. CTN was detected in lipid extracts of all examined samples at concentrations varying between 0.28 and 6.29 microg/g, but was not found in aqueous extracts. When human embryonic kidney cells (HEK293) were incubated for 72 h with Monascus extracts, the concentrations causing 50% cell death by all lipid extracts were in the range of 1.8-4.7 mg/mL, whereas aqueous extracts showed a lower cytotoxicity. Incubation of HEK293 cells with 60 microM pure CTN for 72 h caused cell viability to fall to 50% of control levels. In addition, coadministration of pure CTN and lipid extracts from Monascus samples significantly enhanced CTN cytotoxicity for HEK293 cells using the MTT assay. These results provide the first information about the cytotoxic effects of various Monascus samples and CTN-Monascus mixtures on a human cell line.     

Chromate Generation by Chromate Depleted Subsurface Materials

Soluble chromate concentrations as high as 200 μg Cr L^-1 have been reported in water samples from monitoringwells tapping alluvial deposits allegedly contaminated bylaboratory waste as well as control wells off site andupgradient (shallow aquifer) near Davis California, U.S.A. In this report we present evidence that these Cr(VI) levelscould have been generated by geogenic processes rather thanby anthropogenic inputs. We tested the hypothesis thatnative Cr(III) has been and can be oxidized to chromate bynative manganese oxides. Twenty-three drill core samples(all unsaturated) were retrieved from depths varying from1.5 to 22.5 m in 6 different wells. Visible nodules ofMnO₂ were dispersed throughout many of the samples andcarbonates were also present. Sample pH values averagedabout 8.0 and organic C was mostly less than 1.0 g kg^-1. Total Mn and Cr averaged 835 and 191 mg kg^-1respectively. All samples had the capability to oxidizeadded Cr(III) to Cr(VI). To determine the inherent capabilityof the samples to produce Cr(VI) from native Cr(III), subsamples were extracted with 5 mM CaSO₄ plus 5 mM MgSO₄ until Cr(VI) was no longer detected. After freeze-drying, deionized-distilled water was added to theleached samples to approximately field capacity (0.03 MPa). Freeze drying did not generate Cr(VI). These samples wereincubated in polyethylene film bags at room temperature inthe dark. After 1 week incubation, water in the samples wasextracted by centrifugation and the extracts were analyzedfor Cr(VI). All of the samples generated Cr(VI), and theconcentrations in the extracts ranged from 20 to 100 μg Cr L^-1. Total chromium, endemic chromium VI and chromium VI generated in leached samples were not statistically different between samples from onsite and control samples taken offsite and upgradient in respect to the shallowest aquifer.     

Effects of dehulled adlay on the culture count of some microbiota and their metabolism in the gastrointestinal tract of rats

Experiments were conducted to study the effect of a dietary supplement of dehulled adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) on the culture counts of some important groups of intestinal bacteria and their metabolism in the gastrointestinal (GI) tract of Sprague-Dawley rats. Rats were divided into four groups, and each group was fed a diet containing different levels of dehulled adlay for 30 days as follows: 0% (control), 5%, 20%, and 40%. All animals fed with adlay had normal healthy intestinal walls and no pathogenic signs whatsoever. There were no significant differences in body weight gain or the cecal pH between different groups of rats. Both the 20% and 40% groups had lower culture counts of enterics in their feces than the 5% and control groups, whereas the culture counts of fecal lactic acid bacteria were higher in feces of rats fed with adlay than in the control group. Cecal total short-chain fatty acid (SCFA) content and fecal SCFA were significantly higher in the 20% and 40% groups than in the control and 5% groups. All the adlay-fed rats had a higher fecal butyric acid concentration than the control rats. It is concluded that adlay has a significant influence on the growth of intestinal bacteria, which may ultimately affect the physiology and other functions of GI tracts of rats.     

Volatile flavor components in red fermented soybean (Glycine max) curds

Red fermented soybean (Glycine max) curds (FSC) were extracted with a simultaneous steam distillation and extraction (SDE) apparatus and analyzed by gas chromatography-mass spectrometry (GC-MS) and further evaluated by gas chromatography-mass spectrometry-flame ionization detector-olfactometry (GC-MS-FID-O). A combined total of 89 compounds were identified from three samples (A-C). The numbers of components identified (samples A-C) were 87, 85, and 81, respectively, which were divided into 10 compound classes. The quantity of 48 compounds was statistically significant (p < 0.05). Sample B generally had the lowest level, whereas sample C had the higher level of each common component among the samples. GC-MS-FID-O analyses showed that the majority of the fruity, diacetyl, cantaloupe-like odors were eluted early during the run (RI < 1100), followed by the meaty flavor, rosy, and prune-like aromas (RI > 1100). Ethyl 2-methylpropanoate, 2,3-butanedione, ethyl butanoate, ethyl 2-methylbutanoate, 3-(methylthio)propanal, benzeneacetaldehyde, and ethyl 3-phenylpropionate were found to be important common components contributing to the characteristic aroma of red FSCs.