• Title/Summary/Keyword: in vitro screening

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The screening test on the efficacy of anthelmintics by using third-stage larvae and adult of cultivation in vitro (시험관내에서 인공배양한 제 3기 자충 및 성충을 이용한 구충효능 선발시험)

  • Jee, Cha-ho;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.589-594
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    • 1998
  • The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

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Antifungal activities of sulphamide and dicarboximide fungicides against Botrytis cinerea in several in vitro bioassays (여러 종류의 in vitro 생물검정에서 Botrytis cinerea에 대한 sulphamide계와 dicarboximide계 살균제의 활성 특성)

  • Choi, Gyung-Ja;Kim, Heung-Tae;Kim, Jin-Cheol;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.3 no.3
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    • pp.37-44
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    • 1999
  • Two sulphamide (dichlofluanid and tolylfluanid) and three dicarboximide fungicides (iprodione, vinclozolin, procymidone) were used to investigate the correlation between in vitro antifungal activities and in vivo disease controlling activities against Botrytis cinerea, a causal agent of tomato gray mold and to develop efficient in vitro assays. They controlled effectively the development of tomato gray mold disease in vivo and their controlling activities were similar one another. However, several in vitro assays revealed that their in vitro antifungal activities were quite different between sulphamide and dicarboximide fungicides; the formers showed stronger inhibition activities for spore germination than the latters, whereas the formers inhibited mycelial growth less severely than the latters. The results indicate that the fungicides having different modes of action can show different in vitro antifungal activities according to in vitro assays, even if they have similar in vivo disease controlling activities. On the other hand, two rapid and efficient in vitro assays named Microtiter plate methods I (MPM I) and II (MPM II) were developed for the evaluation of fungicides for inhibitory activities against spore germination and mycelial growth of B. cinerea, respectively. The antifungal activities of five fungicides of two chemical groups in MPM I and II were correlated with the inhibitory activities against spore germination and mycelial growth using solid media, respectively.

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Reliability of In Vitro Assay for Initial Depigmenting Agent Screening (미백제 선발을 위한 In Vitro 측정법의 신뢰도)

  • Nguyen, Dung H.;Nguyen, Duc T.M.;La, Lyun Hwa;Lee, Hyang-Bok;Shin, Jung-Hyun;Kim, Eun-Ki
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.183-188
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    • 2008
  • Initial screening assay for depigmenting agents includes in vitro mushroom tyrosinase assay and antioxidant assay. Based on this screening result, melanin synthesis in melanocyte, in screened samples, is further measured. Measuring cellular melanin needs time, human resource, and skills. Therefore initial screening method should be reliable. We examined, 34 Chinese herbs, correlated the screening assay methods with cellular melanin. No reliable relationship was observed between factors, indicating the limitation in the use of these assays, probably due to the complexicity of melanogenesis.

Evaluation of In vitro and In vivo Screening Methods for Estrogenic Activity of Endocrine Disruptors (내분비계 장애물질의 에스트로겐 활성에 대한 In vitro 및 In vivo 검색시험법)

  • 김형식;한순영;한상국;신재호;문현주;김소희;박기숙;김규봉;이이다
    • Toxicological Research
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    • v.16 no.2
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    • pp.109-116
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    • 2000
  • The purposes of our study were to optimize the conditions of the screening and testing methods for endocrine disruptors, to characterize these assays using several compounds with well-defined endocrine activity, and to compare the sensitivity between these assays currently undergoing validation. Two in vitro test systems, MCF-7 cells proliferation (E-screen assay) and competitive binding to estrogen receptors (ER) were selected to evaluate the estrogenic effects. 17$\beta$-Estradiol (E2) and diethylstilbestrol (DES) were used as a positive control in vitro test. Also, E2 and ethinyl estradiol (EE) were used as a positive control in vivo uterotrophic assay. In in vitro test, E2 and DES showed a strong estrogenic response at concentration of 1.0 nM. In uterotrophic assay, E2 (0.3 $\mu\textrm{g}$/kg) and EE (0.3 $\mu\textrm{g}$/kg) produced a significant increase in uterus and vagina weight in both immature and ovariectomized rats. Although we did not com-pared the specificity between in vivo and in vitro assays, these assay systems may serve as a good tool for endocrine disruptors screening methods. Our data indicate that these assay systems exhibit some difference in their sensitivity to the same estrogenic compounds. Therefore, as a first rapid screening assay for estrogenic activity qf unknown chemicals, at least two assay systems should probably be carried out with a view of high sensitivity and standardization conditions. Also, a careful validation tests are necessary to obtain a reasonable degree of reproducibility.

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Implications for the Predictivity of Cell-Based Developmental Toxicity Assays Developed Two Decades Apart

  • Kawamura, Satoshi;Horie, Nobuyuki;Okahashi, Noriko;Higuchi, Hashihiro
    • Toxicological Research
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    • v.35 no.4
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    • pp.343-351
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    • 2019
  • Many in vitro developmental toxicity assays have been proposed over several decades. Since the late 1980s, we have made intermittent attempts to introduce in vitro assays as screening tests for developmental toxicity of inhouse candidate products. Two cell-based assays which were developed two decades apart were intensively studied. One was an assay of inhibitory effects on mouse ascites tumor cell attachment to a concanavalin A-coated plastic sheet surface (MOT assay), which we studied in the early days of assay development. The other was an assay of inhibitory effects on the differentiation of mouse embryonic stem cell to beating heart cells (EST assay), which we assessed more recently. We evaluated the suitability of the assays for screening in-house candidates. The concordance rates with in vivo developmental toxicity were at the 60% level. The EST assay classified chemicals that inhibited cell proliferation as embryo-toxic. Both assays had a significant false positive rate. The assays were generally considered unsuitable for screening the developmental toxicity of our candidate compounds. Recent test systems adopt advanced technologies. Despite such evolution of materials and methods, the concordance rates of the EST and MOT systems were similar. This may suggest that the fundamental predictivity of in vitro developmental toxicity assays has remained basically unchanged for decades. To improve their predictivity, in vitro developmental toxicity assays should be strictly based on elucidated pathogenetic mechanisms of developmental toxicity.

Screening Method for Antihepatotoxic Activity Using $CCl_4-induced$ Cytotoxicity in Primary Cultured Rat Hepatocytes (일차 배양 흰쥐 간세포에서 $CCl_4$ 유발 세포독성을 이용한 간보호 효과 검색방법)

  • Kim, Young-Sook;Park, Ki-Hyun
    • Korean Journal of Pharmacognosy
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    • v.26 no.1
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    • pp.51-56
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    • 1995
  • To devise an in vitro screening method for antihepatotoxic activity, $CCl_4-induced$ cytotoxicities in primary cultures rat hepatocytes were examined. When rat hepatocytes were intoxicated with 0.5, 1.0 or 1.5 mM $CCl_4$ for 1.5, 3 or 19hr, in order of LDH>GOT>GPT release form hepatocytes was increased in a dose-dependent manner. Treatment with 1.5 mM $CCl_4$ for 1.5 hr showed maximum increase in activity of LDH, GOT or GPT released in the medium compared with the control. At this experimental condition, well known antihepatotoxic substances, glycyrrhizin and silybin markedly inhibited $CCl_4-induced$ cytotoxicities. These results demonstrated that the screening method using $CCl_4-induced$ injury in primary cultured rat hepatocytes might be suitable in vitro assay for antihepatotoxic activity.

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Practices of Antitumor Screening Tests for Natural Products (천연항종양성자원 とスクリニングの 실제)

  • 코이치
    • Korean Journal of Plant Resources
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    • v.5 no.2
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    • pp.95-103
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    • 1992
  • Present anticancer drugs in the clinical side have not showed a conclusive effect of the chemotherapy for cancer patients. In order to find much more efficient antitumor agents fromnatural resources, various screening methods vivo and in vitro have been developed by manyresearchers. The intention of this paper is to provide an outline of some background on the tumorsystem in drug development of natural products, to review some screening programs for theevaluation of antitumor activity and to introduce the practical procedures of some antitumorscreening methods in vivo and in vitro. At the end of this paper, the current literatures related toantitumor natural products from higher plants at our laboratory are described.Key words'anticancer drugs, screening methods.

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Anti-Ulcer Activity of Newly Synthesized Acylquinoline Derivatives

  • Cheon, Hyae-Gyeong;Kim, Hyun-Jung;Mo, Hye-Kyoung;Shin, En-Joo;Lee, Yeon-Hee
    • Archives of Pharmacal Research
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    • v.22 no.2
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    • pp.137-142
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    • 1999
  • Anti-ulcer activity of newly synthesized acylquinoline derivatives was investigated. For the in vitro screening, the effects of compounds on gastric $H^{+}/K^{+}$ ATPase isolated from hog and rabbit were examined. Among them, AU-090, AU-091, AU-254, AU-413 and AU-466 exhibited good in vitro activity on both enzymes. To correlate the in vitro activity with in vivo action, the effects of the compounds on the basal gastric acid secretion were studied. Some derivatives showed considerable anti-secretory activities, and AU-413 was selected for further studies. AU-413 protected gastric damage induced by either ethanol or NaOH dose dependently when given orally. $ED_{50}$ values of 12 mg/kg, p.o. (ethanol) and 41 mg/kg, p.o. (NaOH) were obtained. In addition, histamine-stimulated gastric secretion was reduced upon AU-413 administration. Taken together, newly synthesized acylquinoline derivatives, especially AU-413, is worthy of further investigation to be developed as an anti-ulcer agent.

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