Changes in colonic microbiotas in rat after long-term exposure to low dose of okadaic acid.

Okadaic acid (OA), one of the most important phycotoxins, is widely distributed around the world, concerning diarrheic shellfish poisoning (DSP), and even colorectal cancer. Here, we found that long-term exposure of OA at a low dose (80 µg kg-1 body weight) had certain effects on colonic microbiotas and tract in rat. In the OA-exposed rat, colonic epithelium layer was damaged, and relative abundance of some microbiotas were significantly changed, especially genera in Clostridiales. However, no intestinal inflammation or significant disease was observed. Combined with the increase in relative abundance of some genera in Clostridiales induced by OA in the fermentation experiment, we proposed that OA could cause damage to the intestinal epithelium and increase the relative abundance of pathogenic bacteria, thereby increasing the probability of contact between intestinal epithelium and pathogenic bacteria and leading to an easier pathogenicity.

Encapsulation of Vitamin E and Soy Isoflavone Using Spiral Dextrin: Comparative Structural Characterization, Release Kinetics, and Antioxidant Capacity during Simulated Gastrointestinal Tract.

Spiral dextrin subfraction (SD-40) obtained through enzyme debranching and gradient ethanol precipitation could interact with vitamin E (VE) or soy isoflavone (SIO) to form V-type inclusion complexes. The formation of two inclusion complexes was confirmed by Fourier transform-infrared spectroscopy, atomic force microscopy, and differential scanning calorimetry. In this study, an in vitro gastrointestinal model was used to investigate the breakdown of inclusion complexes and release behavior of bioactive compounds. The results indicated that the two inclusion complexes exhibited a controlled and sustained release behavior during digestion. In addition, the SD-40/VE inclusion complex presented higher stability and stronger antioxidant capacity than the SD-40/SIO inclusion complex. Furthermore, the first and zero order models were applied to understand the release kinetics of VE and SIO from inclusion complexes in the stomach, whereas the first order model was chosen to describe the release of VE and SIO from inclusion complexes in the intestine.

Protection of feruloylated oligosaccharides from corn bran against oxidative stress in PC 12 cells.

Feruloylated oligosaccharides (FOs) were prepared by autoclaving corn bran in oxalic acid (0.6%) solution, and their protection effects against oxidative stress in pheochromocytoma cells (PC 12) cells were investigated. The FOs samples, which comprised a mixture of feruloylated mono- and dipentoses with 4.88% bound ferulic acid (FA), as well as xylose, arabinose, galactose, and glucose amounting to 46.43, 40.46, 3.76, and 8.68% of the total sugars, respectively, were prepared by autoclaving the pretreated corn bran in 0.6% oxalic acid and then further separated. Antioxidant activity was tested by 1,1-diphenyl-2-picrylhydrazyl radical 2,2-diphenyl-1-(2,4,6-trinitrophenyl)hydrazyl (DPPH) scavenging and oxygen radical absorbance capacity (ORAC) methods. Oxidative stress was induced by H2O2 in PC 12 neuronal cell culture model. The results showed that FOs exhibited higher antioxidant activity than free ferulic acid, with an IC50 value of 11 versus 128 µM for DPPH and an ORAC value of 4.77 versus 2.62 µmol Trolox/µmol. Tetrazolium blue assay showed that the addition of FOs with an FA concentration >50 µM significantly increased cell viability after treatment with H2O2. Flow cytometry analysis showed that the addition of FOs at concentrations of 800, 200, and 50 µM significantly decreased the apoptosis rate at the sub-G0 phase from 37.5 to 12.7, 16.2, and 20.9% (P < 0.01), respectively. FOs also significantly decreased the malonic dialdehyde content and lactate dehydrogenase (LDH) activity, but increased superoxide dismutase activity in PC 12 cells treated with H2O2 and prevented the damage of cellular membranes by decreasing the release of LDH to the cultures. The addition of FA at 800 µM showed an effect similar to that of FOs at 200 µM. Therefore, the FOs prepared from corn bran are potential functional ingredients for protection against oxidative stress.

Reporter gene assay for detection of shellfish toxins.

OBJECTIVE: To explore the potential reporter gene assay for the detection of sodium channel-specific toxins in shellfish as an alternative for screening harmful algal bloom (HAB) toxins, considering the fact that the existing methods including HPLC and bioassay are inappropriate for identifying HAB toxins which poses a serious problem on human health and shellfish industry. METHODS: A reporter plasmid pEGFP-c-fos containing c-fos promoter and EGFP was constructed and transfected into T24 cells using LipofectAMINE 2000. Positive transfectants were screened by G418 to produce a pEGFP-c-fos-T24 cell line. After addition of increasing neurotoxic shellfish poison (NSP) or GTX2,3, primary components of paralytic shellfish poison (PSP), changes in expression of EGFP in the cell line were observed under a laser scanning confocal microscope and quantified with Image-pro Plus software. RESULTS: Dose-dependent changes in the intensity of green fluorescence were observed for NSP in a range from 0 to 10 ng/mL and for GTX2,3 from 0 to 16 ng/mL. CONCLUSION: pEGFP-c-fos-T24 can be applied in detecting HAB toxins, and cell-based assay can be used as an alternative for screening sodium channel-specific HAB toxins.