Effect of Heavy Metals on Embryonic Development in the Mussel, Mytilus galloprovincialis

Sung, Chan-Gyoung,Kim, Gi-Beum,Lee, Chang-Hoon The Malacological Society of Korea 2006 The Korean Journal of Malacology Vol.22 No.2

The embryos of marine bivalves have been commonly used in bioassays for quality assessments of marine environments. Although several standard protocols for the developmental bioassay of bivalves have been proposed, only a few trials for application of these protocols in environmental assessments or for the development of a new protocol with Korean species have been conducted. As such, there is a strong need to establish standard bioassay protocols with bivalves commonly found in Korean waters. To determine the sensitivity of Mytilus galloprovincialis to establish a standard bioassay, their fertilized eggs were exposed to six metals (Ag, Cd, Cr, Cu, Ni, and Zn). The order of biological impact was Ag > Cu > Ni > Zn > Cr > Cd and their lowest observed effective concentration were 5, 16.4, 25.4, 142, 187 and 1,500${\mu}g/l$, respectively. The proportion of normal larvae appeared to decrease linearly with the logarithm of each toxicant concentration within the tested range. The average values of median effective concentrations $(EC_{50})$ from the triplicate experiments for Ag, Cd, Cr, Cu, Ni, and Zn were 6.8, 1,797, 786, 16.6, 68.1, and 139.2${\mu}g/l$, respectively. There was a more than 100-fold difference in $EC_{50}$ values of Cu and Cd. The value of $EC_{50}$ or median lethal concentration of Cu was within the range observed for other bivalve developmental bioassays. The overall sensitivity of M. galloprovincialis in the present developmental bioassay was also similar to that of other marine organisms commonly used in aquatic bioassays (e.g. sea urchins, oysters). Hence, the bioassay using the embryo of M. galloprovincialis is considered to be a useful tool to monitor and evaluate the quality of marine aquatic environments.

식물홀몬 및 생장조절물질의 생물검정기술 II. Abscisic Acid 및 Brassinolide

최충돈 한국작물학회 1989 한국작물학회지 Vol.34 No.1

A bioassay is a test system using a living organism (in whole or in part) to determine the presence or relative potency of chemical substances. The development and uses of bioassay are intimately linked to the discovery and characterization of the major classes of plant hormones. An application of this relationship is helpful for understanding the concept of plant hormones as well as the use of bioassay. And plant bioassay have been development and employed not only for the discovery and characterization of the biological activity of plant growth regulators but also have served several important secondary roles. The ideal bioassay should possess the characteristic of high specificity. great sensitivity. short response time, low cost and ease of obtaining plant material. acceptable ease of manipulation, and minimal space and equipment requirements.

An attempt to organize bioassays' data of various chemicals

Sakoda, Akiyoshi,Shoji, Ryo,Sakai, Yasuyuki,Suzuki, Motoyuki,Utsumi, Hideo 嶺南大學校 環境問題硏究所 1998 環境硏究 Vol.18 No.1

Abstract Environmental waters such as river and lake waters are most likely polluted by a numerous number of chemicals produced and emitted by human activities nowadays. Bioassays are considered to be rational methodologies for evaluating their toxicities against human beings and ecosystems. The various kinds of bioassays data for chemicals have been accumulated so far in the literature. Organizing bioassays data of various chemicals was tried in this work in order to make use of those data much more effectively and practically for the toxicity control and management of environmental waters. Environmental waters such as river and lake waters are most likely polluted by a numerous number of chemicals produced and emitted by human activities nowadays. Bioassays are considered to be rational methodologies for evaluating their toxicities against human beings and ecosystems. The various kinds of bioassays data for chemicals have been accumulated so far in the literature. Organizing bioassays data of various chemicals was tried in this work in order to make use of those data much more effectively and practically for the toxicity control and management of environmental waters.

Development of Luciferase Reporter Gene-based Cell Bioassay for the Aromatic Hydrocarbon Receptor Agonists

Sun-Young Kim,Eun-Jung Choi,Jae-Ho Yang 대한생리학회-대한약리학회 2006 The Korean Journal of Physiology & Pharmacology Vol.10 No.6

The aromatic hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biological and toxicological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related chemicals. The application of recombinant reporter plasmid such as the firefly luciferase gene has proven to be a very effective method to detect these chemicals. The bioassay system, CALUX, is sensitive in directly detecting AhR-agonists from a variety of environmental and biologic materials. However, responses of the AhR-dependent bioassays are dependent on the cell types used. Thus, we developed a sensitive bioassay using the recombinant mouse hepatoma cell (Hepa1c1c7) for the determination of dioxins. The recombinant cell line was stably transfected with firefly luciferase reporter gene (pGudLuc1.1). The transfected cells showed the highest induction of luciferase activity at 4.5 hr and a decrease beyond this time point. The system showed the highest sensitivity of detection ever reported. Upon TCDD exposure cells showed 2 fold increase at 10 pM and 7 fold increase at 100 pM, respectively. The passage number after the transfection played an important role in the sensitivity. The increase of passage number tended to increase the sensitivity of the cells up to 15. The media without phenol red showed a higher induction rate than with phenol red, suggesting the preferable use of phenol red-free media for the bioassay. Since each of the assays has unique characteristics that make them suitable for some screening applications and not others, development of sensitive bioanalytical methods based on a variety of cellular systems in a key to the successful determination of dioxins. The bioassay system developed in this study will contribute to further development of successful screening the AhR agonists among the environmental mixture. In addition, the rapid and sensitive nature of this cellular system can be applied as a valuable tool to screen the dioxin-like moieties among the prodrugs at the initial stage, thereby expediting the new drug discovery.

Development of Luciferase Reporter Gene-based Cell Bioassay for the Aromatic Hydrocarbon Receptor Agonists

Kim, Sun-Young,Choi, Eun-Jung,Yang, Jae-Ho The Korean Society of Pharmacology 2006 The Korean Journal of Physiology & Pharmacology Vol.10 No.6

The aromatic hydrocarbon receptor (AhR) is a ligand-activated transcription factor that mediates many of the biological and toxicological effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, dioxin) and related chemicals. The application of recombinant reporter plasmid such as the firefly luciferase gene has proven to be a very effective method to detect these chemicals. The bioassay system, CALUX, is sensitive in directly detecting AhR-agonists from a variety of environmental and biologic materials. However, responses of the AhR-dependent bioassays are dependent on the cell types used. Thus, we developed a sensitive bioassay using the recombinant mouse hepatoma cell (Hepa1c1c7) for the determination of dioxins. The recombinant cell line was stably transfected with firefly luciferase reporter gene (pGudLuc1.1). The transfected cells showed the highest induction of luciferase activity at 4.5 hr and a decrease beyond this time point. The system showed the highest sensitivity of detection ever reported. Upon TCDD exposure cells showed 2 fold increase at 10 pM and 7 fold increase at 100 pM, respectively. The passage number after the transfection played an important role in the sensitivity. The increase of passage number tended to increase the sensitivity of the cells up to 15. The media without phenol red showed a higher induction rate than with phenol red, suggesting the preferable use of phenol red-free media for the bioassay. Since each of the assays has unique characteristics that make them suitable for some screening applications and not others, development of sensitive bioanalytical methods based on a variety of cellular systems in a key to the successful determination of dioxins. The bioassay system developed in this study will contribute to further development of successful screening the AhR agonists among the environmental mixture. In addition, the rapid and sensitive nature of this cellular system can be applied as a valuable tool to screen the dioxin-like moieties among the prodrugs at the initial stage, thereby expediting the new drug discovery.

토양, 종자 및 유묘 검정법에 의한 전남지역 수집 여뀌바늘의 ALS 저해 제초제 Penoxsulam에 대한 저항성 수준

장세지,이도원,정장용,윤영범,이도진,이정란,권오도,국용인 한국잔디학회 2019 Weed & Turfgrass Science Vol.8 No.2

This study was carried out to investigate the resistant levels of Ludwigia prostrata to acetolactate synthase (ALS) inhibiting herbicides through soil, seed and whole plant bioassays. A total of 358 soil samples were collected from Gwangju and Chonnam provinces. We also collected seed samples from 26 different Ludwigia prostrata plants from the same areas. We conducted soil tests on collected samples to determine resistance levels to imazosulfuron+pyriminobac-methyl in Ludwigia prostrata. We also conducted seed and whole plant tests from collected species to determine resistance levels to penoxsulam. In the soil tests, Ludwigia prostrata resistance to imazosulfuron+pyriminobac-methyl was confirmed in 12 of the 358 samples (3.3%). Notably, resistance rates of Ludwigia prostrata was the highest in soils from Goheung-gun with 20%. In the seed bioassay, Ludwigia prostrata resistance to penoxsulam was confirmed in 12 of the 26 collecting areas (46.1%). Also worth noting, Ludwigia prostrata from Jangheung (4) was 4 times more resistant to penoxsulam than standard susceptible plants of the same species. However, in whole plant bioassay, Ludwigia prostrata resistance to penoxsulam was confirmed in all 26 collecting areas. In particular, Ludwigia prostrata of Boseong (7) and Jangheung (1) showed appoximately 13 times higher resistance to penoxsulam than standard susceptible plants of the same species. Therefore, it is suggested that we should develop a control system that can inhibit the spread of Ludwigia prostrata which is resistant to penoxsulam and to control it effectively in rice field. 본 연구는 토양, 종자 및 유묘검정법에 의해 ALS 저해 제초제에 여뀌바늘 저항성 수준을 알아보기 위하여 수행되었다. 본 연구를 위해 광주광역시를 포함한 전남지역에서 총 358 지점으로부터 토양을 채취하였고, 26개 지점으로부터 여뀌바늘 종자를 수집하였다. 토양검정은 imazosulfuron+pyriminobac-methyl을, 종자 및 유묘검정은 penoxsulam를 사용하였다. 토양검정에서 imazosulfuron+pyriminobac-methyl 저항성 여뀌바늘은 총 358지점 중 12지점(3.3%)에서 확인되었다. 특히 저항성 여뀌바늘 발생률은 고흥군이 20%로 가장 높게 나타났다. Seed bioassay법에 의한 penoxsulam 저항성 여뀌바늘은 26개 지역 중 12지역(46.1%)에서 확인되었다. 특히 장흥(4)의 여뀌바늘 수집종은 표준 감수성에 비해 penoxsulam에4배 저항성을 보였다. 그러나 whole plant bioassay법에 의해서는 26개 지역에서 penoxsulam에 저항성으로 나타났다. 특히 보성(7)과 장흥(1) 여뀌바늘 수집종은 표준감수성에 비해 약 13배 이상의 높은 저항성을 보였다. 따라서 벼 재배지에서 penoxsulam 저항성 여뀌바늘 확산을 억제하고 이를 효율적으로 방제할 수 있는 방제 체계를 개발해야 할 것으로 사료되었다.

식물홀몬 및 생장조절물질의 생물검정기술 I. 옥신, 지베렐린 및 싸이토키닌

이정명 한국작물학회 1989 한국작물학회지 Vol.34 No.1

식물홀몬을 포함한 각종 식물생장조절물질의 생검법(bioassays or biological assays)을 소개 및 비교검토함과 동시에 일관성 있는 결과를 얻기 위한 몇가지 유의점을 지적코자 옥신류 생검에서는 녹두발근과 귀리제1절간장신장, gibberellin에서는 왜성벼의 제2엽신장과 왜성완두의 상조축신장, 그리고 cytokinin류에서는 무자엽생장, 담배줄기수캘러스생장, 그리고 오이자엽내엽록소형성 등의 방법을 요약하였다. 아울러 일련의 실험을 통해 얻어졌던 각종생장조절물질(특히 최근에 개발되는 각종 왜화제)에 대한 반응을 홀몬별로 요약 및 비교하였다. The objective of this paper is to compare and summarize the procedure and effectiveness of some bioassay systems and to point out ways to obtain reliable results from each bioassay. Detailed C:escriptions were given for those widely-adapted bioassay methods, such as mungbean rooting (auxin), Avena first internode straight growth (auxin), dwarf rice growth (gibberellin), dwarf pea epicotyl elongation (gibberellin), radish cotyledon expansion test (cytokinin), and tobacco stem pith callus growth (cytokinin), and the effects of various plant growth regulators including some recently introduced growth retardants (Paclobutrazol, Uniconazol, etc.) were also summarized.

사람치아 단백질을 분리 흡착한 PVDF막의 생체반응에 관한 연구

강나라(Nara Kang),홍종락(Jong-Rak Hong),정필훈(Pill-Hoon Choung) 대한구강악안면외과학회 2004 대한구강악안면외과학회지 Vol.30 No.3

Purpose: Human tooth proteins are highly heterogeneous, comprising diverse proteins derived from a number of genes. The attempts to identify protein for activity of tooth matrix proteins have been defied by several factors. First, the amount of proteins within teeth is very small relative to many extracellular matrix proteins of other tissues. Second, the bioassay system is tedious and needed for long time. Therefore we tried to find easy techniques, which increase the product rate, and an assay of small proteins, with which amino acid sequence is possible without additional procedures. Materials and Methods: Total protein were extracted from 300 g enamel removed teeth and 600 g teeth with 4 mol/L guanidine HCl and purified by gel chromatography. Aliquot of proteins was implanted into muscle pouches in Sprague-Dawley rats for bioassay. By SDS-PAGE and membrane blotting, molecular weight of each protein was estimated and a partial amino acid sequence was obtained. Each fraction blotted on the membrane was cut out and inserted in rat ectopic model. Results: In dissociative method, total tooth proteins were obtained 1mg/ml from enamel removed teeth and 3.5 mg/ml from teeth. In SDS-PAGE, four clear bands at the sites corresponding to 66, 40, 20 and 18 kD. Especially The 66 kD band was clearly exhibited. Amino acid sequencing from tooth could be possible using PVDF membrane blotting technique. In amino acid sequencing, 66 kD protein was identified as albumin. Conclusion: Compared with conventional method for extraction of teeth protein and bioassay of proteins, the methods in this study were easy, time-saving and more productive technique. The matured tooth proteins omitting additional procedure of mechanical removal of enamel were simply analyzed using blotted PVDF membrane. This method seems to make a contribution as a technique for bioassay and amino acid sequencing of protein.

식물홀몬 및 생장조절물질의 생물검정기술 2 : Abscisic Acid 및 Brassinolide

崔忠惇 韓國作物學會 1989 한국작물학회지 Vol.34 No.S

A bioassay is a test system using a living organism (in whole or in part) to determine the presence or relative potency of chemical substances. The development and uses of bioassay are intimately linked to the discovery and characterization of the major classes of plant hormones. An application of this relationship is helpful for understanding the concept of plant hormones as well as the use of bioassay. And plant bioassay have been development and employed not only for the discovery and characterization of the biological activity of plant growth regulators but also have served several important secondary roles. The ideal bioassay should possess the characteristic of high specificity. great sensitivity. short response time, low cost and ease of obtaining plant material. acceptable ease of manipulation, and minimal space and equipment requirements.

지중해담치, Mytilus galloprovincialis의 배 발생에 미치는 다환방향족탄화수소류 (2-methylnaphthalene, fluorene, dibenzothiophene, phenanthrene, pyrene) 의 영향

성찬경,박판수,이종현 한국패류학회 2014 The Korean Journal of Malacology Vol.30 No.3

Mussels have been commonly used in bioassay for quality assessments of environment. Moreover, several standard protocols for the developmental bioassay of bivalves have been proposed. In this study, the EC50 of polycyclic aromatic hydrocarbons (PAHs) was determined using mussel, Mytilus galloprovincialis embryonic developmental bioassay. To determine the sensitivity of M. galloprovincialis, their fertilized eggs were exposed to five PAHs (2-metylnaphthalene, fluorene, dibenzothiophene, phenanthrene, pyrene). The EC50 of 2-metylnaphthalene, fluorene, dibenzothiophene, phenanthrene, and pyrene were 232, 273, 67.9, 43.2, and 33.1 μg/L, respectively. The overall sensitivity of M. galloprovincialis in the present developmental bioassay was similar to or more sensitive than that of other marine organisms commonly used in aquatic bioassays. The results of this study could be provide with fundamental data of setting standard for protection of marine life and or can use prediction the aquatic toxicity of PAHs.