RESUMO
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.
Assuntos
Poluentes Químicos da Água , Purificação da Água , Esgotos , Poluentes Químicos da Água/análise , Estrogênios , Arilsulfatases , GlucuronidaseRESUMO
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.
Assuntos
Gastroenterite/tratamento farmacológico , Fatores Imunológicos/farmacologia , Ostreidae , Peptídeos/farmacologia , Animais , Organismos Aquáticos , Ciclofosfamida , Citocinas/metabolismo , Modelos Animais de Doenças , Gastroenterite/induzido quimicamente , Gastroenterite/microbiologia , Microbioma Gastrointestinal/efeitos dos fármacos , Imunomodulação/efeitos dos fármacos , Imunossupressores , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Fitoterapia , Organismos Livres de Patógenos EspecíficosRESUMO
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.
RESUMO
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.
Assuntos
Anticorpos Monoclonais/biossíntese , Antígeno Ca-125/genética , Proteínas de Membrana/genética , Proteínas Recombinantes/biossíntese , Animais , Anticorpos Monoclonais/imunologia , Especificidade de Anticorpos , Western Blotting , Antígeno Ca-125/biossíntese , Antígeno Ca-125/imunologia , Antígeno Ca-125/isolamento & purificação , Linhagem Celular Tumoral , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Humanos , Proteínas de Membrana/biossíntese , Proteínas de Membrana/imunologia , Proteínas de Membrana/isolamento & purificação , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
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.
Assuntos
Antioxidantes/farmacologia , Flavonoides/farmacologia , Sequestradores de Radicais Livres/farmacologia , Nicotiana/química , Polissacarídeos/farmacologia , Antioxidantes/química , Ácido Ascórbico/análise , Ácido Ascórbico/química , Ácido Ascórbico/metabolismo , Benzotiazóis/metabolismo , Compostos de Bifenilo/metabolismo , Flavonoides/química , Sequestradores de Radicais Livres/análise , Radicais Livres , Radical Hidroxila/metabolismo , Oxirredução , Picratos/metabolismo , Extratos Vegetais/química , Folhas de Planta/química , Polissacarídeos/química , Ácidos Sulfônicos/metabolismo , Superóxidos/metabolismoRESUMO
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.