• Title/Summary/Keyword: yeast-based bioassay

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Modification of Estrogenic Effect of Nonylphenol Combined with DEHP in Yeast-based Bioassay (형질전환효모를 이용한 내분비계장애물질검색과 Nonylphenol의 Estrogen 유사작용에 대한 DEHP의 상협작용)

  • 박미선;정해관;박현신;한의식;김종원;엄미옥;정상희;오혜영
    • Toxicological Research
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    • v.17 no.1
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    • pp.65-71
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    • 2001
  • The key targets of endocrine disruptors are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. A yeast-based steroid hormone receptor gene trascription assay was previously developed for the evaluation of chemicals with endocrine modulating activity. The yeast transformants used in this assay contain the human estrogen receptor along with the appropriate steroid response elements upstream of the $\beta$-galactosidase reporter gene. We tried to evaluate several natural and synthetic steroids of their potential to interact directly with the steroid receptor. Some putative endocrine disruptors, including nonylphenol, are weakly estrogenic. But the combined treatment oj these chemicals with di-(2-ethylhexyl)phthalate (DEHP) significantly increased the $\beta$-galactosidase activity in the yeast transformant. These results suggest that we also have to consider the synergistic effects of endocrine disruptors. In this study, we showed that yeast-based bioassay is a valuable tool for screening potential endocrine disruptors and quantitative determination of estrogenicity. And the possibility that the estrogen receptor binds multiple environmental chemicals adds another level of complexity to the interaction between the endocrine disruptors and the human hormone system.

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Multiple-end-point Bioassays Using Microorganisms

  • Iwahashi, Hitoshi
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.6
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    • pp.400-406
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    • 2000
  • Since the 1950s, the numbers of species and chemicals produced have significantly increased. Despite the fact that industrial chemicals have given us numerous benefits, there is no doubt that they have damaged the environment. The chemicals being dispersed on the earth should be carefully controlled to prevent adverse effects. Bioassay is one of the methods to assess chemical safety. In bioassay systems, chemical safety is estimated by monitoring biological responses to environmental pollutants and newly synthesized chemicals. This report introduces multiple-point bioassay systems that are based on chemical sensitivities of microorganisms, responses of one kind of organism, and micro-array technology. Multiple-end-point bioassays enable the prediction of chemicals in the environment and the understanding of toxicities of newly synthesized chemicals.

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A Bioassay of Ginseng Extract s Based on Yeast Growth Determination (효모성장 측정을 이용한 인삼추출물의 생물학적 검정)

  • Jung, Noh-Pal
    • Journal of Ginseng Research
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    • v.5 no.1
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    • pp.24-34
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    • 1981
  • For bioassay of the various extracts of ginseng, the growth determination method using Saccharomyces cerevisiae which was cultured with various doses of the extracts, was studied The water extract, Powder. and ethanol extract were more effective (about 45∼ 110% increase) than saponins or its fractions (about 20∼35% increase). The cold methanol residue showed a increase effect but it was not significant. The bioassay curves for the water extract, ethanol extract, the butanol extracted saponins and the cold methanol- residue were made from the experimental data. From these curves it is possible to find the relation between dose and effectiveness and the optimal doses of various ginseng extracts, and the amount of extract in a sample can be estimated The .angers of sample amount were 0.01% (100ppm) ∼0.32% (3200ppm) fo. the water extract, 0.025% (150ppm)∼0.1% (1000ppm) for the ethanol extract, and 0.008% (80ppm)∼0.016% (160ppm) for the saponins. It was impossible to determine the range for the cold methanol- residue, The acceleration effects on the cell proliferation by a only 0.0008% (8ppm) of the diol- and triol-saponin were measurable in earlier Period (24 hour treatment).

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Comparison of Short-Term Toxicity Tests Based on Feeding Behavior and Temperature Control by Ceriodaphnia dubia (Ceriodaphnia dubia의 먹이섭생 기작과 온도조절에 근거한 급성독성조사법의 비교)

  • Park, Jong-Ho;Lee, Sang-Ill;Cho, Young-Oak
    • Journal of Korean Society on Water Environment
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    • v.20 no.1
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    • pp.48-54
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    • 2004
  • Two methods, a Ceriodaphnia algal uptake suppression test (CAUST) and a new toxicity test based on temperature control (TTBTC) which are based on feeding behaviour and temperature control, respectively, were developed and compared for the adoption as the better methodology for short-term toxicity screening. As previously published by Lee et aI., (1997), the CAUST method is based on the feeding behaviour of C. dubia and requires as little as 1 hour of contact time between C. dubia neonates and toxicant. However, even though CAUST requires only 1 hour of contact time, this method still take many hours for the preparation and measurement. Before the test starts, neonate digestive tracts were cleared by feeding yeast to the daphnids, Neonates were then exposed to toxicant, followed by addition of Scenedesmus subspiatus into the bioassay vessels. Daphnids were examined under the bright-field microscope with the presence of algae (indicated by a green colored digestive tract) or the absence of algae. Uptake indicated no toxic effect, whereas, absence of uptake indicated toxic inhibition. Unlike CAUST, the newly developed method (TTBTC) is based on just temperature control for the toxicity test of C. dubia. Initially, neonates are exposed to toxicants while the temperature of water bath containing media increased to $35.5^{\circ}C$. After 1.25 hour of contact time, the number of the daphnids, either live (no toxic effect) or dead (toxic effect), is counted without the aid of any instrument. In both methods, median effective concentrations ($EC_{50}$ values) were computed based on the results over a range of dosed toxicant concentrations. It showed that TTBTC was as sensitive as the standard 48-hour acute bioassay and CAUST. TTBTC and CAUST were much more sensitive than the I-hour I.Q. test and 30-minute Microtox. This study indicates that TTBTC is an easier and more rapid toxicity test than the standard 48-hour acute bioassay and even CAUST.

Detection of estrogenic hormone 17β-estradiol in soil samples by a recombinant yeast bioassay and supercritical fluid extraction

  • Shim, Jae-Han;Kim, Mi-Ra;Topp, Edward;Choi, Jeong-Heui;Mamun, Iqbal Rouf
    • Korean Journal of Environmental Agriculture
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    • v.27 no.4
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    • pp.447-455
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    • 2008
  • Recombinant yeast estrogenicity (YES) assay was used as a bioanalytical tool in order to screen $17{\beta}$-estradiol in the soil samples collected from different sites of South Korea. Solvent extraction and supercritical fluid extraction (SFE) methods were compared for the extraction of the estradiol from the soils. Most high detection of the estradiol based on YES assay was observed in the soils extracted with methanol. Different types of estrogenic hormones including $17{\beta}$-estradiol were suggested to be possibly exiting in the soils, since the methanol extracts of the soils showed an estrogenic activity that was not observed in the hexane extracts of the soil. SFE extracts showed estrogenic activity in some of the samples but methanol extract showed best activity.

In vitro Screening of Medicinal Plants with Estrogen Receptor Modulation Activity (생약의 여성호르몬 수용체 조절 활성 검색)

  • Lee, Chang-Min;Kang, Se-Chan;Oh, Joa-Sub;Choi, Han;Li, Xue-Mei;Lee, Jae-Hyun;Lee, Mi-Hyun;Choung, Eui-Su;Kawk, Joung-Hwan;Zee, Ok-Pyo
    • Korean Journal of Pharmacognosy
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    • v.37 no.1 s.144
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    • pp.21-27
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    • 2006
  • Yeast based estrogenicity assay is the simplest and useful for the assay and the discovery of novel estrogenic substances in natural specimens, The estrogen receptor(ER) modulation activity of 50% EtOH extracts of 101 traditional medicinal herbs was assessed using a recombinant yeast assay system with both a human estrogen receptor expression plasmid and a receptor plasmid. Among them, 14 species proved to be active. Pureariae Flos (flower of Puerraria thunbergiana BENTH.) had the highest estrogenic relative potency$(7.75{\times}10^{-3})$ $(EC_{50}=9.39\;{\mu}g/ml)$. The $EC_{50}$ value of $17{\beta}-estradiol$ used as the positive control was $0.073\;{\mu}g/ml)$ (Relative Potency=1.00). There results demonstrated that some of the traditional medical herb may be useful in the therapy of estrogen replacement.

Toxicity Evaluation of Endocrine Disrupting Chemicals Using Human HepG2 Cell Line, Lumbricus rubellus and Saccharomyces cerevisiae (HepG2 인간 세포주, Lumbricus rubellus 및 Saccharomyces cerevisiae를 이용한 내분비교란물질의 독성평가)

  • Sohn, Ho-Yong;Kim, Hong-Ju;Kum, Eun-Joo;Cho, Min-Seop;Lee, Jung-Bok;Kim, Jong-Sik;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.16 no.6
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    • pp.919-924
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    • 2006
  • Toxicity evaluation systems for various chemicals and their metabolites have been developed during last decades. In this study, the acute toxicity of endocrine disrupting chemicals, such as endosulfan, bisphenol A, vinclozolin, and 3,5-dichloroaniline, was evaluated using HepG2 cell line, Lumbricus rubellus and Saccharomyces cerevisiae, respectively. The extents of toxicity of the chemicals in different bioassay systems varied substantially, such as endosulfan>3,5-dichloroaniline> bisphenol A in HepG2 cell line system, endosulfan>bisphenol A>3,5-dichloro aniline in L. rubellus system, and 3,5-dichloroaniline>endosulfan>bisphenol A in S. cerevisiae system. Meanwhile, no cytotoxicity was observed by treatment of vinclozolin in the evaluation systems. Our results suggest that earthworm and yeast are useful to evaluate acute toxicity of endocrine disrupting chemicals, and direct comparison of toxicity data from different bioassay systems is unattainable. Based on our results, we propose that the bioassay system with earthworm or yeast, a rapid, simple and economic system, could be applied as pre-test for the toxicity evaluation using human cell line or animals.

Plant Growth Promoting Effect and Antifungal Activity of Bacillus subtilis S37-2 (Bacillus subtilis S37-2 균주의 항진균활성 및 식물생육촉진 효과)

  • Kwon, Jang-Sik;Weon, Hang-Yeon;Suh, Jang-Sun;Kim, Wan-Gyu;Jang, Kab-Yeul;Noh, Hyung-Jun
    • Korean Journal of Soil Science and Fertilizer
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    • v.40 no.6
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    • pp.447-453
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    • 2007
  • With a broad objective for the development of microbial based fertilizers, a total of 373 strains were isolated from rhizoplane and rhizosphere of pepper, tomato, lettuce, pasture, and grass. The efficacy of the isolates to augument overall plant growth was evaluated. After screening for their plant growth promotion and antagonistic properties in vitro efficient strains were further selected. The most efficient strains was characterized by 16S rRNA gene sequences and biochemical techniques and was designated as Bacillus subtilis S37-2. The strains facilitated plant growth and inhibited the plant phathogenic fungi such as Fusarium oxysporum (KACC 40037, Rhizoctonia solani (KACC 40140), and Sclerotinia sclerotiorum (KACC 40457). Pot based bioassay using lettuce as test plant was conducted by inoculating suspension ($10^5$ to $10^8cells\;mL^{-1}$) of B. subtilis S37-2 to the rhizosphere of lettuce cultivated in soil pots. Compared with non-inoculated pots, marked increase in leaf (42.3%) and root mass (48.7%) was observed in the inoculation group where the 50ml of cell mixture ($8.7{\times}10^8cells\;ml^{-1}$) was applied to the rhizosphere of letuce either once or twice. Antagonistic effects of B. subtilis S37-2 strain on S. sclerotiorum (KACC 40457) were tested. All the tested lettuce plants perished after 9 days in treatment containing only S. sclerotiorum, but only 17% of lettuce was perished in the inoculation plot. B. subtilis grew well in the TSB culture medium. The isolates grew better in yeast extracts than peptone and tryptone as nitrogen source. The growth rate was 2~4 times greater at $37^{\circ}C$ as compared with $30^{\circ}C$ incubation temperature. B. subitlis S37-2 produced $0.1{\mu}g\;ml^{-1}$ of IAA (indole 3-acetic acid) in the TSB medium containing L-tryptophan($20mg\;L^{-1}$) in 24 hours.