• Title/Summary/Keyword: bioassay system

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A Novel Bioassay System for Screening of Compounds Affecting Anthocyanin Biosynthesis Pathway in White Corn Leaf Segment (백화옥수수 잎절편을 이용한 안토시아닌 생합성 조절제 탐색용 신규 검정법 확립)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Lee, Jung-Ae;Oh, Kwang-Hoon;Cho, Kwang-Yun
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.207-214
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    • 2003
  • This study was carried out to establish a novel bioassay system for screening of compounds affecting aromatic amino acid or anthocyanin biosynthesis through investigating a degree of sucrose-induced anthocyanin formation such as size of plant material, buffer conditions, light intensity and irradiated duration, incubation temp., etc were determined and standard procedure (suitable experimental condition) was set up as follows. The second leaf blade of white corn seeding induced by fluridone treatment were segmented into a size of 5${\times}$5 min. The segments were floated on the solution of 1% sucrose in 1.0mM MES buffer (pH6.0∼6.5) and incubated at 26$^{\circ}C$ for 2days under the continuous light condition(70∼100$\mu$mol m$^{-2}$ s$^{-1}$ ). Anthocyanin in the purpled tissues was extracted with methanol containing 1% HCl and the optical density of the clear supematants was determined at 528mm. Influences of some chemicals were tested using this system. Glyphosate, 5-enolpyruvylshikimate 3-phosphate synthase inhibitor, showed most sensitive response with I$_{50}$ value at 3.3$\mu$M. Dicyclohexylcarbodiimide(DCCD) and parachloromercuribenzenesulfonic acid(PCMBS) had a relatively strong ingibition with I50 value at 7.1$\mu$M and 10.2$\mu$M, respectively. These results show that sucrose-induced anthocyanin formation in white com leaf segment provide a very simple and rapid system for searching new compounds affecting aromatic amino acid or anteocyanin biosynthesis by screening at less than 10$\mu$M.

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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Nutrient Limitation of Phytoplankton in the Dongbok Lake: Analyses of Long-term data and Bioassay Experiments (동복호 식물플랑크톤의 제한 영양염 규명: 장기 자료 분석 및 생물검정조사)

  • Jeong, Byong-Kwan;Shin, Yong-Sik;Jang, Na-Mik;Kim, Sang-Don
    • Korean Journal of Ecology and Environment
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    • v.41 no.3
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    • pp.412-421
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    • 2008
  • Nutrient limitation was examined for the Dongbak Lake based on bioassay experiments and analyses of long-term monitoring data. From the results of data analyses, molar ratios of TN/TP and DIN/DIP were higher than 16 : 1 suggesting that phytoplankton growth was potentially limited by phosphorus. Phytoplankton responded to the addition of phosphates in the incubation experiments for all seasons whereas phytoplankton did not respond to other nutrients such as ammonia, nitrate and silicates. Size fraction (net and nano size) of phytoplankton also responded to the addition of phosphorus indicating that phytoplankton growth was limited by phosphorus in the Dongbok Lake. There was also a taxonomic shift from euglenophyceae to bacillariophyceae after addition of phosphate during warm season especially.

A bioassay system for pharmacological standardization of Withania somnifera derived herbal remedies

  • Dey, Amitabha;Chatterjee, Shyam Sunder;Kumar, Vikas
    • CELLMED
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    • v.9 no.1
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    • pp.1.1-1.13
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    • 2019
  • Background: Contents of bioactive substances extractable from different parts of terrestrial plants vary enormously. Aim: To ascertain that parts of Withania somnifera other than its roots can also be used for prevention and cure of unavoidable stress triggered central hypersensitivity to pain. Material and Methods: Groups of male or female mice treated either with Withania somnifera extracts or with metformin, aspirin, imipramine, diazepam and niacin for 11 consecutive days were subjected to "foot-shock stress-induced hyperthermia" and "hot plate" tests on the 1st, 5th, 7th, and 10th days of the experiments. On the 11th day, they were subjected to tail suspension test and on 12th day pentobarbital hypnosis test. Results: Except for diazepam and imipramine, protective effects of all other tested drugs as well as of the Withania somnifera extracts against stress-induced central hypersensitivity to pain were accompanied by their preventive effects against foot-shock stress-induced body weight losses. All observed stress response suppressing effects of all test agents increased with increasing numbers of treatment days. However, mean duration of pentobarbital-induced sleep was shorter in the extracts treated groups and longer in the diazepam treated ones only. Conclusions: Reported observations reveal that pharmacological activity profile of Withania somnifera extracts in male and female mice are almost identical, and are not like those of several drugs currently often prescribed for the treatment of diabetes-associated comorbidities. Withanolides are not the only extractable bioactive constituents of Withania somnifera. The described bioassay system is well suited for pharmacological standardization of diverse types of Withania somnifera extracts.

Biological Evaluation of off-shore Water Quality in Southern Coastal Waters with Asterias Aurensis Gametes (아므르불가사리(Asterias amurensis)의 배우자를 이용한 남해해역 연안해수의 생물학적 수질평가)

  • 유춘만;박지인;조기안;주현수;박종천;나명석
    • Journal of Environmental Science International
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    • v.8 no.1
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    • pp.1-6
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    • 1999
  • This study was carried out to evaluate off-shore water quality and to draw marine pollution map to Southern sea in Korea by bioassay using gametes and early development system of Asterias amurensis. From the bioassay, it was determined that the water qualites of Southern sea maintained the grade II, because development of A. amurensis were moderately inhibited. However, Sunchon, Kwangyang, and Jinhae Bay belonged to grade III, because development of A. amurensis were strongly inhibited. This could be due to the inflows of several river systems and poor water circulation. The water quality at Yosu island was the grade I, because A. amurensis were not inhibited in its early development.

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Establishment of Foliar Application Assays for Developing Natural Herbicides (천연물 제초제 개발을 위한 전식물체 수준의 경엽처리 검정법 확립)

  • Kim, Jae-Deog;Jang, Hyun-Woo;Seo, Bo-Ram;Hwang, Hyun-Jin;Choi, Jung-Sup;Kim, Jin-Seog
    • Korean Journal of Weed Science
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    • v.30 no.2
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    • pp.153-163
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    • 2010
  • This study was carried out to establish an improved bioassay system, whole-plant bioassay which is more effective in developing natural herbicides for foliar treatment such as herbicidal essential oils. Two bioassay systems using four weed species (Echinochloa crus-galli, Digitaria sanguinalis, Aeschynomene indica, and Abutilon theophrasti), spraying method and spotting method, were established. Spraying method is applicable if the amount of test compounds is enough, while spotting method is useful for the small amount of test compounds. The initial application rate was desirable at $2,500{\sim}5,000\;{\mu}g\;mL^{-1}$. Herbicidal activities were higher in the NOP treatment when compared to the Tween 20 treatment. To efficiently evaluate volatile compounds such as essential oils, if the compound-treated pots were incubated in dew chamber for about 10hrs, better results were obtained in the degree and stability of herbicidal responses. When the efficiency of bioassay systems established in this study was compared, the spraying method was minimized four times to the conventional method that has beed used for screening of synthetic compounds in KRICT. On the other hand, in the spotting method, screening for development of a natural herbicides was possible even in level of 1/100 test volume and 1/200 amounts of test compound compared to the spraying method.

Evaluating Limiting Nutrients through Long-term Data Analyses and Bioassay Experiments in Cheonsu Bay and Taean Sea (장기자료 분석과 생물검정실험을 이용한 천수만과 태안해역의 제한영양염 평가)

  • Kim, Jin Hyun;Jeong, Won Ok;Shin, Yongsik;Jeong, Byungkwan
    • Journal of the Korean Society of Marine Environment & Safety
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    • v.28 no.4
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    • pp.459-468
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    • 2022
  • Long-term data analyses and bioassay experiments were conducted to assess limiting nutrients in Cheonsu Bay and Taean sea. First, long-term nutrient data (2004-2016) provided by the National Water Quality Monitoring Network were used to assess potential limiting nutrients. Analysis of the long-term data showed that the dissolved inorganic nitrogen/dissolved inorganic phosphate (DIN/DIP) ratio was mostly below 16, with N limitation being dominant. A subsequent analysis using the concentration ratios of N, P, and Si showed that N limitation was dominant during summer and autumn but that Si limitation occasionally occurred during winter and spring in relatively limited areas. However, the dominant limiting nutrient was not determined. The nutrient analysis of the field water collected during the bioassay experiment showed that DIN/DIP revealed P limitation at all stations in March and May, whereas N limitation was dominant in July and October. In the analysis using the concentration ratios of N, P, and Si, P and Si limitation appeared in March and May, but there were points with no dominant limiting nutrient. However, N limitation was dominant in July and October. In the bioassay experiment for assessment of the actual limiting nutrient, the results showed no specific limiting nutrient in March, whereas NH4+ and NO3- showed responses in May, July, and October, which confirmed that N was a substantial limiting nutrient directly involved in phytoplankton growth during this period.

The Identification of Limiting Nutrients Using Algal Bioassay Experiments (ABEs) in Boryeong Reservoir after the Construction of Water Tunnel

  • Ku, Yeonah;Lim, Byung Jin;Yoon, Jo-Hee;Lee, Sang-Jae;An, Kwang-Guk
    • Korean Journal of Environmental Biology
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    • v.36 no.4
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    • pp.558-566
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    • 2018
  • The objective of the study was to determine nutrition regime and limitation in the Boryeng Reservoir where there's a water tunnel between Geum River and the reservoir. Evaluation was conducted through in situ algal bioassay experiments (in situ ABEs) using the cubitainer setting in the reservoirs. For in situ ABEs, we compared and analyzed variations in chlorophyll-a (CHL-a) and phosphorus concentrations in Boryeong Reservoir before and after the water tunnel construction. We then analyzed the nutrient effects on the reservoir. Analysis for nitrogen and phosphorus was done in the three locations of the reservoir and two locations of the ABEs. The in situ ABEs results showed that phosphorous and Nitrogen, the primary limiting nutrient regulating the algal biomass was not limited in the system. The treatments of phosphorus or simultaneous treatments of N+P showed greater algal growth than in the control of nitrate-treatments, indicating a phosphorus deficiency on the phytoplankton growth in the system. The water from the Geum River had 5 times higher total phosphorus (TP) than the water in the reservoir. Efficient management is required as pumping of the river water from Geum River may accelerate the eutrophication of the reservoir.

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.

In Vitro Bioassay for Transforming Growth Factor-$\beta$ Using XTT Method

  • Kim, Mi-Sung;Ahn, Seong-Min;Moon, Aree
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.903-909
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    • 2002
  • Research in the cytokine field has grown exponentially in recent years, and the validity of such studies relies heavily on the appropriate measurement of levels of cytokines in various biological samples. Transforming growth factor (TGF)-$\beta$, a hormonally active polypeptide found in normal and transformed tissue, is a potent regulator of cell growth and differentiation. The most widely used bioassay for TGF-$\beta$ is the inhibition of the proliferation of mink lung epithelial cells. Though detection of [$^3$H]thymidine incorporation is more sensitive than the MTT assay, it presents some disadvantages due to the safety and disposal problems associated with radioisotopes. In this study, we attempted to ascertain the experimental conditions which could be used for measuring the in vitro biological activity of TGF-$\beta$ in a safer and more sensitive way compared with the currently available methods. We compared the commonly used method, the MTT assay, to the XTT assay using different parameters including cell number, incubation time and the wave length used for detecting the product. We examined the anti-proliferative activities of TGF-$\beta$ in three different cell lines: Mv-1-Lu mink lung epithelial cells, MCF10A human breast epithelial cells and H-ras-transformed MCF10A cells. Herein, we present an experimental protocol which provides the most sensitive method of quantifying the biological activity of TGF-$\beta$, with a detection limit of as low as 10 pg/ml: Mv-1-Lu or H-ras MCF10A cells ($1{\times}10^5/well$) were incubated with TGF-$\beta$ at $37^{\circ}C$ in a humidified $CO_2$ incubator for 24 hr followed by XTT treatment and determination of absorbance at 450 or 490 nm. Our results may contribute to the establishment of an in vitro bioassay system, which could be used for the satisfactory quantitation of TGF-$\beta$.