• Title/Summary/Keyword: Bioassay methods

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Establishment of CALUX Bioassay for Dioxin Determination

  • Joung, Ki-Eun;Chung, Young-Hee;Sheen, Yhun-Yhong
    • Environmental Mutagens and Carcinogens
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    • v.24 no.3
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    • pp.137-142
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    • 2004
  • Dioxin-like compounds are ubiquitous environmental polltants that could be accumulated in biological system and toxic to human and wildlife. Given this issue, it is important to develop a reliable dioxin detection methods for a rational risk assesment of dioxin-like compounds. In this study, we tried to set up and validate a sensitive, reliable risk assessment of dioxin-like compounds. In this study, we tried to set up and validate a sensitive, reliable and rapid bioassay model, CALUX bioassay as a screening tool for routine measurement of dioxin-like conpounds in environmental matrices. For the valisation of CALUX bioassay, firstly, we performed dose-response assay for 2,3,7,8-TCDD, most potent dioxin-like compound, using two different methods CALUX and EROD assay. Induction of luciferase activity and CYPIA catalyzed EROD activity were dose-dependently induced by 2,3,7,8-TCDD, with initial induction at 0.1 pM and maximal induction at 1 nM. In order t determine whether the CALUX bioassay could predict the effects of dioxin-like compounds, 2,3,7,8-TCDD dose-response from CALUX was compared with that from EROD assay. The correlation coefficient ($r^2$) was found to be 0.89, indicating a good correlation between two different methods and the possibility of CALUX bioassay as a useful dioxin detecting method.

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Establishment of Bioassay System for Developing New Insecticides II. Differences in Susceptibilities of the Insect Species to Insecticides according to Different Application Methods (살충제 개발을 위한 생물검정법의 확립 II. 처리방법 및 종간에 따른 살충제 감수성 차이)

  • 안용준;김길하;박노중;조광연
    • Korean journal of applied entomology
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    • v.31 no.4
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    • pp.452-460
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    • 1992
  • To establish the economical and reliable routine bioassay system for developing new insecticidal compounds, effects of leaf-dipping time, application methods, insect species and their developmental stages on susceptibilities of insects to insecticides were studied. The stable insecticidal activity appeared at the dipping time for 30-60 seconds in leaf-dipping method, and the most effective application methods were leaf-dipping method for apterous green peach aphid adults, and third instars of diamond-back moth and tobacco cutworm, whereas seedling+insect spray method for adults or third instars of brown planthoppers. For two-spotted spider mite, leaf-dipping or intact plant spray method was favorable. In the bioassay for chitin synthesis inhibitors, the inoculation of third instars of brown planthopper, diamond-back moth, tobacco cutworm and green peach aphid, and larvae of two-spotted spider mite to the young host plants treated by spray method were adequate bioassay methods.

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Developing Noninformative Priors for Parallel-Line Bioassay

  • Kim, YeongHwa;Heo, JungEun
    • Communications for Statistical Applications and Methods
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    • v.9 no.2
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    • pp.401-410
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    • 2002
  • This paper revisits parallel-line bioassay problem, from a Bayesian point of view using noninformative priors such as Jeffreys' prior, reference priors, and probability matching priors. After finding the orthogonal transformation, the class of first order and second order probability matching priors are derived. Jeffreys' prior and reference priors are derived also. Numerical examples are given to show the effectiveness of noninformative priors.

A Preliminary Study for Development of a Bioassay Protocol Using the Sperm of a Starfish, Asterias amurensis

  • Ryu, Tae-Kwon;Lee, Chang-Hoon;Park, Jin-Woo
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.10a
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    • pp.158-158
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    • 2003
  • Bioassays using gametes of sea urchins are widely used in ecotoxicological assessments of marine environments. Since most of sea urchin species in Korean coastal water spawn from spring to autumn, bioassay with them during the winter is impossible. In the course of developing standard methods for bioassays with Korean species, we found a winter-spawning starfish, Asterias amurensis, Since reproductive mode of asteroids is similar to echinoids, the bioassay protocol for sea urchins could be applied similarly to the starfish. Here, we tested and determined several conditions for the acceptability of bioassay with A. amurensis. The least required time for formation of fertilization membrane of fertilized eggs to be easily distinguished from unfertilized ones was 60 min. The threshold of sperm to egg ratio that could make acceptable fertilization rates in controls was 3000. The allowed time for manipulation of sperm after dilution in seawater was at most 3 hr. The optimal exposure time of sperms when the response against toxicant solution was relatively stable was in the range of 20-60 min. The tolerance range of sperms to the salinity of test solution was 26-38 psu. The sensitivity of A. amurensis sperm was intermediate among marine organisms commonly used in aquatic toxicity tests. The sperm bioassay with A. amurensis can be satisfactorily applied to toxicity assessments of marine environments.

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BIOASSAY OF HUMNA TOOTH PROTEIN BLOTTED POLYVINYLIDENE DIFLUORIDE(PVDF)MEMBRANE (사람치아 단백질을 분리 흡착한 PVDF막의 생체반응에 관한 연구)

  • Kang, Na-Ra;Hong, Jong-Rak;Choung, Pill-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.3
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    • pp.186-192
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    • 2004
  • 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.

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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Development History of Pregnancy Test Technology (임신진단검사기술의 발전사)

  • Kwon, Young-Il
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.4
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    • pp.382-390
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    • 2018
  • The history of the pregnancy test started in ancient Egypt with a germination test using wheat and barley. Since then, unscientific methods have been used from the days of Hippocrates and Galen to the Middle Ages when uroscopy was used, even until the early 1800s. On the other hand, since the mid-1800s, scientific methods and evidence have begun to emerge, which led to scientific research on the physiological characteristics of pregnancy. The first attempt to utilize these scientific findings was initiated with the bioassay by Aschheim and Zondek using rats and mice in 1927, and then developed into experiments using rabbits and frogs. The immunoassay method, which started in the 1960s, contributed greatly to the generalization of the pregnancy tests while improving the problems of the bioassay. In 1976, a pregnancy test kit was introduced that can be used at home, contributing to the popularization of pregnancy tests. Since the 1980s, technological advances in diagnostic tests have also been applied to pregnancy tests to further improve the reliability of the diagnosis of pregnancy. In the 2000s, the accuracy and ease of use of the pregnancy test kits for home use have improved drastically. This study examined the history and scientific development of the pregnancy test.

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 Journal of Physiology and Pharmacology
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    • v.10 no.6
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    • pp.349-354
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    • 2006
  • 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.

A Bayesian Analysis in Multivariate Bioassay and Multivariate Calibration

  • Park, Nae-Hyun;Lee, Suk-Hoon
    • Journal of the Korean Statistical Society
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    • v.19 no.1
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    • pp.71-79
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    • 1990
  • In the linear model which consider both the multivariate parallel-line bioassay and the multivariate linear calibration, this paper presents a Bayesian procedure which is an extension of Hunter and Lamboy (1981) and has several advantages compared with the non Bayesian techniques. Based on the methods of this article we discuss the effect of multivariate calibration and give a numerical example.

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Comparison of MBA and HPLC Post-column Oxidation Methods for the Quantification of Paralytic Shellfish Poisoning Toxins

  • Yu, Hongsik;Lim, Keun Sik;Song, Ki Cheol;Lee, Ka Jeong;Lee, Mi Ae;Kim, Ji Hoe
    • Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.159-164
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    • 2013
  • The mouse bioassay and high performance liquid chromatography (HPLC) post-column oxidation method are different methods of quantifying paralytic shellfish poisoning toxins. In this study, we compared their ability to accurately quantify the toxicity levels in two types of field sample (oysters and mussels) with different toxin profiles for routine regulatory monitoring. A total of 72 samples were analyzed by both methods, 44 of which gave negative results, with readings under the limit of detection of the mouse bioassay ($40{\mu}g/100g$ saxitoxin [STX] eq). In 14 oysters, the major toxin components were gonyautoxin (GTX) 1, -2, -3, -4, -5, decarbamoylgonyautoxin-2 (dcGTX2), and decarbamoylsaxitoxin (dcSTX), while 14 mussels tested positive for dcSTX, GTX2, -3, -4, -5, dcGTX2, neosaxitoxin (NEO), STX, and dcSTX. When the results obtained by both methods were compared in two matrices, a better correlation ($r^2=0.9478$) was obtained for mussels than for oysters ($r^2=0.8244$). Additional studies are therefore needed in oysters to investigate the differences in the results obtained by both methods. Importantly, some samples with toxin levels around the legal limit gave inconsistent results using HPLC-based techniques, which could have a strong economic impact due to enforced harvest area closure. It should therefore be determined if all paralytic shellfish poisoning toxins can be quantified accurately by HPLC, and if the uncertainties of the method lead to doubts regarding regulatory limits.