Cytochrome P4501B1(CYP1B1) is known to be inducible by xenobiotic compounds such as policyclic aromatic hydrocarbon(PAH) and dioxins such as 2,3,7,8-tetrachloro-dibenzo-p-dioxin(TCDD). And these induction of CYP1B1 is also regulated by many categories of chemicals. In order to investigate the effects of several chemicals on CYP1B1 gene expression in Hepa-I and MCF-7 cells, 5' flanking DNA of human CYP1B1 was cloned into pGL3 basic vector containing luciferase gene, and then transfected into these cells. After treatment of chemicals, the luciferase activity was measured. CYP1B1 enzyme metabolize PAHs and estradiol. CYP1B1 metabolize estradiol to 4-hydrozyestradiol that is considered as carcinogenic metabolite. Recent industrialized industrialized society, human has been widely been exposed to widespread environmental contaminants such as PAHs(polycyclic aromatic hydrocarbon) that are originated from the imcomplete combustion of hydrocarbons. PAHs are known to be ligands of the AhR(aryl hydrocarbon receptor). Induction of cytochrome P4501B1(CYP1B1) in cell culture is widely used as a biomarker for PAHs. Therefore we have studied the effect of PAHs in the human breast cancer cells MCF-7 to evaluate bioactivity of PAHs. We have used the United State of America EPA selected 13 different PAHs, PAHs mixtures and extracts from environmental samples to evaluate the bioassay system. We examined effects of PAHs on the CYP1B1-luciferase reporter gene and CYP1B1 mRNA level. Benzo(k)fluoranthene and dibenzo(a, h)anthracene showed strong response to CYP1B1 promoter activity stimulation, and also CYP1B1 mRNAs increase in MCF-7 cells in a concentration-dependent manner. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1B1 mRNA. Some flavonoids such as genistein, daidzein, chrysin, naringenin and morin were also investigeted. These flavonoids decreased B(k)F infuced luciferase activity at low concentration. But, these flavonoids exhibited stimulatory effect at high concentration.
Journal of the Korean Society of Food Science and Nutrition
/
v.37
no.2
/
pp.141-147
/
2008
This study investigated the isoflavone contents and physiological properties of non-fermented soybean (NF) and the fermented soybeans prepared with Asp. oryzae (AO) and B. natto (BN). The total isoflavone contents (daidzin, genistin, daidzein and genistein) of NF, AO and BN were 81.8 mg/100 g, 130.7 mg/100 g and 139.5 mg/100 g, respectively. Especially, the total phenol contents of NF, AO and BN were 2.1%, 4.3% and 7.6%, and the total flavonoid contents were 1.3%, 1.6% and 2.7%, respectively. The nitrite-scavenging abilities of NF, AO and BN were 34.4%, 55.2% and 92.5%, respectively. Antimicrobial activity of BN was shown to be the strongest to Bacillus cereus, Staphylococcus aureus and Escherichia coli. The SOD-like activity was the strongest in AO, whereas the electron donating ability was the strongest in BN. Antioxidant activity of AO at concentration of 0.02% was stronger than BN or NF.
Korean traditional soy paste (Doenjang) was fermented using Meju prepared by the culture of wild microorganisms in steamed soy beans. During the fermentation, changes in the physicochemical and several functional properties were monitored. Total acidity and amino acidity increased from 0.09 to 0.96, and 2.24% to 3.28% respectively, Amylase and protease activities increased and showed the maximal level after 60 days of fermentation, which were 4.03 and 7.29 units/ml, respectively. However, both enzyme activities decreased after then. The inhibitory activities against tyrosinase and angiotensin converting enzyme increased and reached 20.57 and 38.18% respectively. Antimutagenic activities against N-methyl-N'-nitro-N-nitiosoguanidine and 4-nitro-O-phenylenediamine (NPD) increased for 90 days and roached 70.21 and 60.01% in S. enterica serovar Typhimurium TA100, respectively. Against NPD and 4-nitroquinoline-1-oxide, the antimutagenic activities also increased and reached 50.91 and 46.35% in the strain TA98, respectively.
Transcriptional activation of the embryonic genome initiates at 2-cell stage in mouse embryo and is characterized by the synthesis of TRC which is restricted to 2-cell stage. To investigate the roles of various protein kinases on the embryonic gene activation, the effects of protein kinase inhibitors on in vitro development and protein synthetic profiles of the early mouse embryos were examinded. None of ${\alpna}-amanitin$ which is a mRNA synthetic inhibitor, H8 which is a PKA inhibitor, and H7 which is a PKC inhibitor, affected on first cleavage of mouse 1-cell embryos in vitro. However, all of these drugs inhibited the second cleavage. When the drugs were removed following treatment for 6 hours, H8 or H7 treatment showed little inhibition on subsequent development of 1-cell embryos to 2-cell stage or further. In contrast, ${\alpna}-amanitin$ irreversibly inhibited the development of 1-cell embryos to 2-cell stage following removal of the drug. Genistein, a TPK inhibitor, inhibited both the first cleavage of 1-cell embryos and the second cleavage of 2-cell embryos, suggesting that TPK activity may be important during the early cleavages. All of the above four drugs inhibited TRC synthesis as shown by the fluorographic analysis of $[^{35}S]-Met$ labeled protein profiles. When late 1-cell embryos were treated with H7 and analyzed synthetic patterns of $[^{35}S]-Met$ labeled protein, the quantitative differences of protein synthesis on SDS-PAGE appeared on 77 kD and 33 kD region at $32{\sim}38$ hours post hCG. From these studies, transcriptional activation of embryonic genome is not essenting to the mouse 1-cell embryos to develop to 2-cell stage. Hawever, TPK activity is reguisite for both the first cleavage and second cleavage. Similarly, both PKC and PKA activities are required for the second cleavage of mouse embryos, but not for the first cleavage.
Background: Endocrine disruptors are exogenous substance, interfere with the endocrine system, and disrupt hormonal functions. However, the effect of endocrine disruptors in different species has not yet been elucidated. Therefore, we investigated the possible effects of $17{\beta}$-estradiol (E2), progesterone (P4), genistein (GEN) and 4-tert-octylphenol (OP), on capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. In this in vitro trial, spermatozoa were incubated with $0.001-100{\mu}M$ of each chemical either 15 or 30 min and then assessed capacitation status using chlortetracycline staining. Results: E2 significantly increased capacitation and the acrosome reaction after 30 min, while the acrosome reaction after 15 min incubation in mouse spermatozoa. Simultaneously, capacitation and the acrosome reaction were induced after 15 and 30 min incubation in porcine spermatozoa, respectively. Capacitation was increased in porcine spermatozoa after 15 min incubation at the lowest concentration, while the acrosome reaction was increased in mouse spermatozoa after 30 min (P < 0.05). E2 significantly increased the acrosome reaction in porcine spermatozoa, but only at the highest concentration examined (P < 0.05). P4 significantly increased the acrosome reaction in bovine and mouse spermatozoa treated for 15 min (P < 0.05). The same treatment significantly increased capacitation in porcine spermatozoa (P < 0.05). P4 significantly increased capacitation in mouse spermatozoa treated for 30 min (P < 0.05). GEN significantly increased the acrosome reaction in porcine spermatozoa treated for 15 and 30 min and in mouse spermatozoa treated for 30 min (P < 0.05). OP significantly increased the acrosome reaction in mouse spermatozoa after 15 min (P < 0.05). Besides, when spermatozoa were incubated for 30 min, capacitation and the acrosome reaction were higher than 15 min incubation in E2 or GEN. Furthermore, the responsiveness of bovine, mouse and porcine spermatozoa to each chemical differed. Conclusions: In conclusion, all chemicals studied effectively increased capacitation and the acrosome reaction in bovine, mouse, and porcine spermatozoa. Also we found that both E2 and P4 were more potent than environmental estrogens in altering sperm function. Porcine and mouse spermatozoa were more responsive than bovine spermatozoa.
This study was conducted to investigate the quality characteristics of tofu prepared with addition of cowpea meal (CPM). Any significant difference of L-value between control CPM-O and treatment CPM-I, CPM-II and CPM-III could not be seen as addition ratio of CPM increases (p < 0.05). The b-value of CPM-III (75%) has shown the least value of 14.26, and it has decreased as addition ratio of CPM increases. The result of texture profile did not show any significant difference between control CPM-O and CPM-Iexcept in the case of cohesiveness. The pH range of treatments was 6.36~6.52, which showed no significant difference from controls. The values of turbidity were 1.46~1.81, which had increasing trend as addition ratio of CPM increases. The total bacterial count of CPM-III was $4.40{\times}10^2CFU/g$, which showed significant difference from the control. No Esterichia coli was detected in the control and the treatments. The result of sensory analysis of the treatment CPM-I did not show any significant difference from the control, which implied the treatment CPM-Iwas found to be suitable at the tofu making using byproducts of cowpea-jelly making. Isoflavone glycoside (IG) was contained more on the order of genistin, daidzin, and glycitin, and the content decreased significantly as addition ratio of CPM increases. The content of isoflavone-aglycone (IA) was less than IG, but showed similar behavior of addition ratio of CPM.
Lee Kyung-Hee;Ryu Seung-Hee;Lee Young-Soon;Kim Young-Man;Moon Gap-Soon
Korean journal of food and cookery science
/
v.21
no.2
s.86
/
pp.163-170
/
2005
Soybean is an important plant as the source of protein and oil, as well as phytochemicals such as genistien, daidzein, phenolic acids, phytic acid, tocopherol, and saponin. Chungkukjang, a fermented soybean paste, is common in Korean meals and bacillus is usually used in the fermentation of steamed soybean. For its processing, whole soybeans are boiled in water until the beans are soft, and then the drained beans are wrapped with rice straw or starter and set in a warm place at $65^{\circ}C\;for\;3\;\~\;4\;days$. Normally, the remaining cooked water which was drained from the steamed beans is discarded. We supposed that this water possesses hish amounts of useful components, and we therefore developed a modified method using the cooked water. After fermentation, we added the remaining cooked water which had been drained from the beans to the fermented soy beans and boiled them together. To investigate the bio-functionality of the modified Chungkukjang, the total antioxidative activity, isoflavones contents, phenolic acids, and 3-deoxyglucosone (3-DG) were measured at each stage of the preparation of chungkukjang. The original and modified chungkukjang possessed a high antioxidative activity compared with the other samples, as did the drained water after steaming of the soybean. The contents of genistein, daidzein, and phenolic acids, which contained antioxidative activity, were also increased in the original chungkukjang and their contents were similar in the modified chungkukjang. The content of 3-DG was increased in the modified chungkukjang compare with the original. It is suggested that the active soybean components delivered to the drained water during the steaming process were useful for increasing the bio-functionality of the modified chungkukjang.
The proliferative effects of thirty Oriental medicinal herbs on MCF-7 (estrogen-sensitive breast cancer cell line) and ROS 17/2.8 osteoblast-like cells were determined using the MTT assay. Methanol extracts from several herbs was found to show proliferative activity on the above two cell lines in the range of 5 to 100 $\mu$g/mL. Among these active herbs, the methanol extract from the rhizomes of Drynaria fortunei showed the most potent proliferative activity, and the cell proliferations were significantly increase by 136 and 158% in the MCF-7 and ROS 17/2.8 cells, respectively, when treated with 100 $\mu$ g/mL. Through a bioassay-guided separation, eight flavonoids, including four new flavan-3-ols and two propelargonidins, together with the known (-)-epiafzelechin and naringin, were isolated. Their chemical structures were characterized as (-)-epiafzelechin (1), (-)-epiafzelechin-3-O-$\beta$-D-allopyranoside (2), (-)-epiafzelechin-3-O-(6"-O-acetyl)-$\beta$-D-allopyranoside (3), 4$\beta$-carboxymethyl-(-)-epiafzelechin methyl ester (4), 4$\beta$-car-boxymethyl-(-)-epiafzelechin sodium salt (5), naringin (6), (-)-epiafzelechin-(4$\beta$\rightarrow8)-4$\beta$-car-boxymethylepiafzelechin methyl ester (7) and (-)-epiafzelechin-($4\beta\rightarrow8, 2\beta\rightarrowΟ\rightarrow7)-epiafzelechin-(4\beta\righarrow8)-epiafzelechin (8) by extensive 1D and 2D NMR spectroscopy. Most of these flavonoids, in the range of $10^{-15}∼10^{-6}$ M, accelerated the proliferation of MCF-7 cell, with compounds 7 and 8, in the range of $10^{-15}∼10^{-12}$ M, showing especially potent proliferation effects. Meanwhile, seven flavonoids, with the exception of compound 4, stimulated the proliferation of ROS 17/2.8 cells in the range of $10^{-15}∼10^{-6}$ M, with compounds 5-8 especially accelerating the proliferation, in dose-dependent manners ($10^{-15}∼10^{-9}$ M), and their proliferative effect was much stronger than that of $E_2$ and genistein. These results suggest that propelargonidin dimers and trimers isolated from the rhizomes of Drynaria fortunei may be useful as potential phytoestrogens, which play important physiological roles in the prevention of postmenopausal osteoporosis.
To evaluate changes in functional characteristics of traditional Doenjang during aging, Doenjang was prepared using an extract of Phellinus linteus (Phellinus extract). Control Doenjang was aged without the extract. The protease activity of Doenjang prepared with Phellinus extract was 3.15 units/mL. Tyrosinase and acetylcholinesterase inhibition activities were 45.78% and 55.18% of control, respectively, in the treated sample. When Salmonella enterica serovar Typhimurium TA100 was used as a reporter strain, antimutagenic activities against the mutagens MNNG and NPD were 90.42% and 82.57% of control values in the treated sample. When S. enterica serovar Typhimurium TA98 was used, antimutagenic activities were 60.28% and 50.33% of control, respectively. Hydrogen-donating activity was 86.65% in the treated sample, which was higher than that of the control (61.69%). Daidzin (an isoflavon glucoside) levels in Doenjang prepared with Phellinus extract were higher, by 35.49 mg/kg, than the control, whereas genistin was not detected in either group. Daidzin and genistin aglycone levels were 263.01 mg/kg and 262.60 mg/kg in the control and test groups, respectively.
Cheongkookjang that was prepared with three kinds of soybeans [non-germinated soybean (NG), soybeans germinated for 12 hr (GS12), and soybeans germinated for 24 hr (GS24)] were investigated. The changes in the pH, total aerobes, and slime content of Cheongkookjangs that were prepared with NG, GS12 and GS24 did not significantly differ during their fermentation for 48 hr at $40^{\circ}C$. The total aerobes of the Cheongkookjang variants reached $10^8{\sim}10^9$ CFU/mL after theirfermentation for 48 hr. The total polyphenol content and DPPH-radical-scavenging activities the germionated and non-germinated soybeans did not significantly differ, but increased significantly according to the germination degree during the fermentation. The isoflavone content of the Cheongkookjang with the germinated soybean increased. The isoflavone content of Cheongkookjang variant were 0.141 mg/g (NG), 0.369 mg/g (GS12) and 0.569 mg/g (GS24); their free amino acid contents were 254.26 mg% (NG), 337.49 mg% (GS12) and 528.78 mg% (GS24); and their sensory characteristics such as their taste, color, flavor, bitter taste, texture, and overall acceptability did not significantly differ.
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