• Title/Summary/Keyword: ${\beta}-galactosidase$ assay

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Endocrine Disrupting Activity of Seven Phthalate Analogues in vitro

  • Ryu, Jae-Chun;Kim, Hyung-Tae;Kim, Youn-Jung;Jeon, Hee-Kyung
    • Environmental Mutagens and Carcinogens
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    • v.22 no.4
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    • pp.259-265
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    • 2002
  • Phthalate analogues are a plasticizer and solvent used in industry. Phthalates were reported to be a potential carcinogen classified in the category of suspected endocrine disruptors. Most common human exposure to these compounds may occur with contaminated food. They may migrate into food from plastic wrap or may enter food from general environmental contamination. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of phthalates that possibly threaten the public health. Concern about their use has been mounting. To screen and elucidate the endocrine disrupting activity and their mechanism of phthalate analogues, first of all, E-screen assay was performed in MCF7 human breast cancer cells with seven phthalate analogues. In this cell proliferation assay, only dibutyl phthalate (DBP) showed weak estrogenic activity. Also the yeast-based transcription assay to assess the interactions of DBP with the estrogen, androgen, and progesterone receptors was conducted. DBP in the concentration ranges from 10$^{-16}$ to 10$^{-11}$ M was active in the estrogen transcriptional assay, but it did not show the effect on $\beta$-galactosidase activity in the progesterone and androgen transcriptional assays. These data indicate that DBP shows estrogenic potential and can be classified as weak and/or suspected endocrine disrupting chemicals.

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Direct Analysis of the Transcription of Escherichia coli rnpB Gene Harbored in a Multicopy Plasmid during Bacterial Growth

  • Park, Jeong-Won;Jung, Young-Hwan;Park, Bo-Hyun;Jeoung, Yeon-Hee;Lee, Young-Hoon
    • BMB Reports
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    • v.29 no.3
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    • pp.221-224
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    • 1996
  • To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

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Evaluation of Safety with Gamgung-tang Using Ames and umu Assays (Ames 및 umu assay를 이용한 감궁탕의 안전성평가)

  • Shon Yun Hee;Kim Cheorl Ho;Nam Kyung Soo
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.215-219
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    • 2005
  • Gamgung-tang (GGT) that is included in Gamdu-tang (consists of Glycyrrhizae Radix, black beans) and Gunggui-tang(consists of Angelicae Radix and Cnidii Rhizoma) showed therapeutic effect of autoimmume thyroiditis in the previous reports. GGT was tested for the safety using Ames and umu gene expression mutagenicity tests. In Ames test, Salmonella typhimurium TA98 and TA100 were used to identify mutagenic property, and the number of histidine revertants was measured. In SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a test strain, and we monitored the levels of umu operon expression by measuring the $\beta-galactosidase$ activity. Mutagenic activity in any assays we tested was not found. After treating S-9 mixture with GGT, mutagenic activity was also not found. The results of this study suggested that there was no DNA damage and mutagenicity of GGT.

Skin permeability of the biosynthetic galactosylated 2-phenoxyethanol (생합성된 galactosylated 2-phenoxyethanol의 피부 투과도)

  • Su-Hong Kim;Sun-Beom Kwon;Jun-Sub Kim;Gi-Seong Moon;Kyung-hwan Jung;Hyang-Yeol Lee
    • Journal of the Korean Applied Science and Technology
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    • v.41 no.1
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    • pp.1-8
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    • 2024
  • The safety of cosmetic ingredients is considered paramount. In order to enhance safety, a novel preservative, PE-gal, was bio-synthesized by utilizing the Escherichia coli enzyme 𝛽-galactosidase on the conventional preservative 2-phenoxyethanold (PE). The skin absorption of the bio-synthesized product, PE-gal, intended for use in cosmetics, was evaluated for permeability using the Franz Diffusion Cell Assay system, comparing it with the conventinal preservative PE. When using samples of the same mass concentration, the Flux and Kp values of PE increased over time, indicating a gradual increase in permeability. However, PE-gal did not exhibit sufficient permeability to measure. This suggests that the skin permeability of PE is higher than that of the PE-gal saccharide. According to Marzulli et al., when confirming the degree of permeation using Kp values, the permeation rate of PE was measured as "slow" at a concentration of 1mg/mL. Thus, the transdermal permeability of the divedened form of PE-gal was significantly lower compared to PE.

Assay of Antimutagenic Activities of Vegetable Plants (식용식물(食用植物)의 항변이원성(抗變異原性)에 관한 연구(硏究))

  • Seo, Jung-Sook;Lee, Young-Wook;Suh, Nan-Joo;Chang, Il-Moo
    • Korean Journal of Pharmacognosy
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    • v.21 no.1
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    • pp.88-91
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    • 1990
  • Potential antimutagenic activities of vegetable plants were investigated. 24 vegetables which are frequently consumed by Korean people were extracted with 70% ethyl alcohol to prepare the extract samples. Then those samples were added to the culture media containing mitomycin C($O.3\;{\mu}g/ml$) and the SOS-Chromotest was performed. Positive control, mitomycin C alone, showed about 330 units of ${\beta}$-galactosidase activities. Among 24 vegetable samples, Saxifraga oblougifolia Nakai (취나물, Saxifragaceae) and Platycodon grandiflorum A. De Candolle(도라지, Campanulaceae) showed 136 and 155 units, respectively when mitomycin C was treated. These results indicate that Saxifragae Herba and Platycodi Radix possess protecting action from mutagenic activity produced by mitomycin C.

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Studies on enzyme immunoassay for determining progesterone of bovine plasma and its clinical application I. Optimizing double antibody for progesterone in enzyme immunoassay (Enzyme immunoassay(EIA)에 의한 소의 혈중 progesterone 측정과 이의 응용에 관한 연구 I. 이항체(二項體)의 최적조건에 관한 연구)

  • Kang, Chung-boo;Shin, Jong-uk;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.28 no.2
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    • pp.321-325
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    • 1988
  • This experiment was carried out to determine the progesterone concentration of bovine plasma by liquid phase double antibody enzyme immunoassay. The optimum conditions of assay-system, double (first and second) antibody and carrier (normal rabbit serum) were investigated. The optimum dilution rate of first antibody, second antibody and normal rabbit serum was $10{\times}10^3$ to $15{\times}10^3$, 20 and $1{\times}10^3$ times, respectively.

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Evaluation of Safety with Astragali Radix : Ames, Rec and umu Assays (Ames, Rec 및 umu Assay를 이용한 황기의 안전성평가)

  • Shon, Yun-Hee;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.34 no.1 s.132
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    • pp.80-85
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    • 2003
  • Water extract from Astragali Radix (AR) was tested for the safety using Ames, Bacillus subtilis Rec, and umu gene expression mutagenicity tests. Mutagenic activity in any assays we tested was not found. In Ames test, Salmonella typhimurium TA98 and TA 100 were used to identify mutagenic property, and the number of histidine revertants was measured. In the Recassay, Bacillus subtilis ${H-17(Rec^+)\;and\;M-45(Rec^-)}$ strains were used to test DNA damage activity. In the SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a test strain, and we monitored the levels of umu operon expression by measuring the ${\beta}-galactosidase$ activity. From the results, there was no DNA damage and mutagenicity of AR. Hepatotoxicity of AR to female ICR mice was also monitored by the measurements of s-GOT, s-GPT, LDH activities after oral feeding for 15 days. AR was not shown any significant changes of s-GOT, s-GPT and LDH activities in mice sera.

Transcriptional Regulation of Escherichia coli serC-aroA Operon : Further Support for cAMP-Dependent Expression

  • Sa, Jae-Hoon;Park, Soo-Sun;Lim, Chang-Jin
    • BMB Reports
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    • v.28 no.1
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    • pp.21-26
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    • 1995
  • The Escherichia coli mixed-function serC-aroA operon encodes biosynthethic enzymes for unrelated pathways leading to the syntheses of serine and aromatic amino acids. It has been proposed that the operon is expressed in a cAMP-dependent manner. In this work experiments were performed to investigate the cAMP-dependent expression of the operon. Exogenous cAMP increased ${\beta}$-galactosidase synthesis in the $cya^+$ and cya strains harboring the serC-aroA-lac fusion plasmid. This enhancement was more dramatic in the $cya^-$ strain grown in a minimal medium. In a dot blot assay the serC-aroA mRNA content increased in a concentration-dependent pattern after the addition of exogenous cAMP. The activity of phosphoserine aminotransferase, encoded by the serC gene, apparently increased in E. coli cells after the addition of cAMP. All results obtained confirmed that the expression of the E. coli serC-aroA operon is positively regulated by cAMP at the level of transcription.

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Antimicrobial Activity of an Aqueous Extract of Green Tea against Food Putrefactive Microorganisms (변패미생물에 대한 녹차 물추출물의 항균 활성 분석)

  • Shin, Young-Hee;Oh, Byung-Tae;Choi, Sung-Gil;Heo, Ho-Jin;Lee, Seung-Cheol;Cho, Sung-Hwan
    • Food Science and Preservation
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    • v.16 no.3
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    • pp.392-399
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    • 2009
  • Recent studies indicate that green tea may have anticancer, antioxidant, and antihypertensive effects, and aids body weight control and the promotion of various desirable physiological functions. However, few studies have investigated the antimicrobial effects of green tea. We sought to determine the antimicrobial activity of green tea extract against food spoilage microorganisms. The extract showed remarkable antimicrobial effects against a wide spectrum of putrefactive and food spoilage microorganisms when used at concentrations greater than $500{\mu}g/ml$. The extract showed thermal and pH stability in the range of $40{\sim}150^{\circ}C$ and pH 3.11, respectively. Green tea extract seems to be an ideal natural antimicrobial, considering both efficacy and thermal and pH stabilities. Antimicrobial substances in green tea extract were investigated using electron microscopy and a $\beta$-galactosidase assay. The data showed that the extract contains several efficacious materials, and that their activities are not synergistic but are instead independent. Our data indicate that hydrophilic antimicrobial substances in green tea extract might control food spoilage microorganisms owing to perturbation of the microbial cell membrane.

Investigation of Antioxidant Activity of Houttuyniae Herba and its Effect on 5α-reductase Gene Expression in Dermal Papilla Cells (어성초(魚腥草)의 항산화 효능 확인 및 모유두 세포의 5α-reductase 유전자 발현에 미치는 영향)

  • Cho, Nam Joon;Lee, Byeong Kwon;Lee, Woong Hee;Kim, Kee Kwang;Han, Hyo Sang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.31 no.6
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    • pp.356-361
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    • 2017
  • Houttuyniae Herba is widely used as a cosmetic for enhancing hair growth, and study on promoting mouse hair growth has also been reported. However, studies on the effects of the Houttuyniae Herba on dermal papilla (DP) cells, which play an important role in hair growth, are not well known. For this reason, we studied the effect of Houttuyniae Herba on DP cells. The strong antioxidant activity of Houttuyniae Herba was confirmed by ABTS assay. In the MTS assay, cell viability was reduced to 94.5% in DP cells by treatment of 2 mg/ml concentration of Houttuyniae Herb and cytotoxicity was not observed at 1 mg/ml concentration. The mRNA expression levels of Bone morphogenetic pretein (BMP6), fibroblast growth factor 7 (FGF7), FGF10, and ${\beta}$-galactosidase genes, which are involved in hair growth cycle and hair loss induction, were measured by quantitative RT-PCR after Houttuyniae Herbtreatment. Houttuyniae Herb did not significantly affect mRNA expression of BMP6, FGF7, FGF10, and ${\beta}$-catenin, which are important factors for regulating the hair cycle, including type 1 $5{\alpha}$-reductase. However, mRNA expression of type 2 $5{\alpha}$-reductase, the major cause of male hair loss, was significantly reduced to 56.1% by treatment of Houttuyniae Herbtreatment. Taken together, these results suggest that the Houttuyniae Herbtreatment can help to treat lair loss through removing free radicals and suppression of the expression level of type 2 $5{\alpha}$-reductase in DP cells.