Deconjugation potentials of natural estrogen conjugates in sewage and wastewater treatment plant: New insights from model prediction and on-site investigations.

Natural estrogen conjugates play important roles in municipal wastewater treatment plant (WWTP), but their deconjugation potentials are poorly understood. This work is the first to investigate the relationships between the enzyme activities of arylsulfatase/ß-glucuronidase and deconjugation potentials of natural estrogen conjugates. This work led to three important findings. First, the enzyme activity of ß-glucuronidase in sewage is far higher than that of arylsulfatase, while their corresponding activities in activated sludge were similar. Second, a model based on ß-glucuronidase could successfully predict the deconjugation potentials of natural estrogen glucuronide conjugates in sewage. Third, the enzyme activity of arylsulfatase in sewage was too low to lead to evident deconjugation of sulfate conjugates, which means that the deconjugation rate of estrogen sulfates can be regarded as zero. By comparing their theoretical removal based on enzyme activity and on-site investigation, it is reasonable to conclude that reverse deconjugation of estrogen conjugates (i.e., conjugation of natural estrogens to form conjugated estrogens) likely exist in WWTP, which explains well why natural estrogen conjugates cannot be effectively removed in WWTP. Meanwhile, this work provides new insights how to improve the removal performance of WWTP on natural estrogen conjugates. SYNOPSIS: This work is the first to show how arylsulfatase/ß-glucuronidase could affect deconjugation of natural estrogen conjugates and possible way to enhance their removal in wastewater treatment plant.

Ameliorative Effects of Peptides Derived from Oyster (Crassostrea gigas) on Immunomodulatory Function and Gut Microbiota Structure in Cyclophosphamide-Treated Mice.

The intestinal flora is recognized as a significant contributor to the immune system. In this research, the protective effects of oyster peptides on immune regulation and intestinal microbiota were investigated in mice treated with cyclophosphamide. The results showed that oyster peptides restored the indexes of thymus, spleen and liver, stimulated cytokines secretion and promoted the relative mRNA levels of Th1/Th2 cytokines (IL-2, IFN-γ, IL-4 and IL-10). The mRNA levels of Occludin, Claudin-1, ZO-1, and Mucin-2 were up-regulated, and the NF-κB signaling pathway was also activated after oyster peptides administration. Furthermore, oyster peptides treatment reduced the proportion of Firmicutes/Bacteroidetes, increased the relative abundance of Alistipes, Lactobacillus, Rikenell and the content of short-chain fatty acids, and reversed the composition of intestinal microflora similar to that of normal mice. In conclusion, oyster peptides effectively ameliorated cyclophosphamide-induced intestinal damage and modified gut microbiota structure in mice, and might be utilized as a beneficial ingredient in functional foods for immune regulation.

Antimicrobial effect and mechanism of cinnamon oil and gamma radiation on Shewanella putrefaciens.

The aims of this study were to observe the antimicrobial effect and mechanism of cinnamon oil combined with gamma radiation on Shewanella putrefaciens. Gamma radiation increased the antimicrobial activity of cinnamon oil, and the relative radiation sensitivity of gamma radiation on S. putrefaciens was increased by cinnamon oil. Gamma radiation significantly increased the changes of bacterial morphology, intra-adenosine 5'-triphosphate (intra-ATP) and extra-ATP concentrations and pHin value of S. putrefaciens treated cinnamon oil. Although, gamma radiation used alone didn't damage the bacterial morphology and ATP concentrations significantly. Gamma radiation assisted cinnamon oil to damage the cell permeability and integrity of S. putrefaciens, thus the combination of cinnamon oil and gamma radiation showed a better antimicrobial activity than used alone.

Effect of slaughter methods on the quality of Chilean jack mackerel (Trachurus murphyi) during refrigerated storage.

The main objective of this study was to assess the influence of slaughter methods on the quality of Chilean jack mackerel (Trachurus murphyi) during refrigerated storage on board. Fishes were slaughtered by asphyxia in air (AA), asphyxia in ice water (AI) or stunning fish heads (SH), and the rigor mortis, pH, total volatile basic nitrogen (TVB-N), trimethylamine (TMA), 2-thiobarbituric acid reactive substances (TBARS) and sensory properties for the fishes were analyzed. On day 0, Chilean jack mackerel samples of AI group displayed higher pH values than those of AA and SH groups. TVB-N, TMA and TBARS values of all samples increased with the storage time, and these values of AI had a lower increase than AA and SH. Moreover, samples of AI had a better sensory score than AA and SH during storage. It can be concluded that slaughter method of asphyxia in ice water for Chilean jack mackerel exhibit the better efficiency on maintaining the fish quality during refrigerated storage on board.

In vitro antioxidant properties of flavonoids and polysaccharides extract from tobacco (Nicotiana tabacum L.) leaves.

In the present study, antioxidant properties of flavonoids and polysaccharides from tobacco (Nicotiana tabacum L.) leaves were evaluated in several in vitro systems, e.g., scavenging activities on hydroxyl, superoxide anion, 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals, and reducing power. Flavonoids showed much better activity than polysaccharides in scavenging activities on free radicals. When compared to the positive control, ascorbic acid, both showed weaker antioxidant potential. However, flavonoids possessed comparable superoxide anion, DPPH and ABTS radical scavenging abilities to ascorbic acid at high concentration (600 µg/mL). Meanwhile, it was found that flavonoids had prominent effects on the reducing power, which was equivalent to ascorbic acid, and was significantly higher than polysaccharides. These results clearly indicate that flavonoids are effective in scavenging free radicals and have the potential to be powerful antioxidants. Thus, tobacco leaves could be considered as a potential source of natural antioxidants for food, pharmaceutical, cosmetics or nutraceutical industries.

[Expression and purification of mucin 16 and preparation and characterization of anti-mucin 16 monoclonal antibody].

AIM: To generate monoclonal antibodies (mAbs) against mucin 16 using purified recombinant protein of human mucin 16 N terminus with His tag (His-mucin 16N) as the antigen. METHODS: Mucin 16 N terminus was cloned into a prokaryotic expression vector pET-32. His-mucin 16N was then expressed in E.coli and purified by the affinity chromotography. Cell fusion was performed after the BALB/c mice were immunized with the purified His-mucin 16N protein. We screened hybridoma cell strains producing mAbs against mucin 16. The specificity and titer of the antibodies were characterized with ELISA, Western blotting, immunofluorescent and immunohistochemical staining. RESULTS: The recombinant protein of His-mucin 16N was expressed and purified. A few hybridoma cell strains which could secrete specific mAbs against mucin 16 were obtained, and one anti-mucin 16 mAb with good specificity and high titer was selected and purified. The isotype of this anti-mucin 16 mAb was determined as IgG1, which indicated that this anti-mucin 16 mAb could be used for ELISA, Western blotting, immunofluorescent and immunohistochemical staining. The endogenous expression of mucin 16 in various cancer cell lines or tissues was also examined with this anti-mucin 16 antibody by Western blotting and other immunoassays. CONCLUSION: The recombinant protein of His-mucin 16N was expressed and purified successfully, with which we prepared anti-mucin 16 mAb with good specificity and high titer.

Isolation and identification of antioxidant and hyaluronidase inhibitory compounds from Ficus microcarpa L. fil. bark.

The aerial roots and bark of Ficus microcarpa L. fil. have been used as folk herbs for perspiration, alleviating fever, and relieving pain in Okinawa. The methanol extract of its bark showed high antioxidant and potential inhibitory activity against hyaluronidase. It was fractionated into hexane, ethyl acetate, butanol, and water fractions. As the ethyl acetate fraction exhibited the strongest activity, it was selected for further purification by repeated Sephadex LH-20 column chromatography and preparative HPLC. Seven compounds were isolated and identified as protocatechuic acid, chlorogenic acid, methyl chlorogenate, catechin, epicatechin, procyanidin B1, and procyanidin B3 by analysis of ESI-MS, UV, and (1)H- and (13)C-NMR spectra. All isolated compounds showed strong antioxidant activity when tested by all applied methods. Catechin, epicatechin, procyanidin B1, and procyanidin B3 exhibited excellent inhibitory activity against hyaluronidase. The results indicate that the extract of F. microcarpa bark may be utilized as a potential antioxidant and hyaluronidase inhibitor.

Comparative studies on the biochemical characteristics of natural actomyosin isolated from PSE and normal pork.

Biochemical changes of natural actomyosin from fresh pale, soft, exudative (PSE) and normal pork were studied, and the effects of different storage temperatures and different incubation temperature and times on sample superprecipitation, total sulfhydryl (-SH) content, and ATP (adenosine triphosphate) sensitivity were investigated. The results demonstrated that ATPase activity and thermal stability of PSE actomyosin were lower than those of normal pork; and that PSE actomyosin had higher -SH content than that of normal pork at all incubation temperatures and times tested.