• Title/Summary/Keyword: In vitro screening test

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The screening of the mutagenicity and desmutagenicity for cooked pork meats according to cooking conditions (조리 조건을 달리한 가열조리된 육류(돼지고기)의 변이원성 검색과 제어에 관한 연구)

  • 정경숙;구성자
    • Korean journal of food and cookery science
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    • v.18 no.6
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    • pp.716-722
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    • 2002
  • Meat(beef, pork, chicken, duck) were cooked by three kinds of instruments (gas grill. electric grill, microwave oven) and extracted with 80% methanol. These methanol extracts were performed the Ames test, employing S. typhimurium tester strain TA100 (in vitro) and micronucleus test (in vivo). The methanol extract of cooked pork showed high mutagenicity in 5.0mg/plate without S9 mix and induced a higher mutagenicity with S9 mix than without S9 mix at 5 mg/plate. In all kinds of cookery methods, pork extracts showed high mutagenicity according to increase of cookery temperature (200$\^{C}$, 260$\^{C}$ and 320$\^{C}$). The methanol extract of cooked pork by electric grill (at 260$\^{C}$, for 5 min) showed high mutagenicity in all kinds of cookery instruments on the Ames test and micronucleus test. In all kinds of meat, the methanol extract of cooked pork showed a higher mutagenicity than the others and chicken showed a lower. The extract after pork soaked in ginger juice showed lower mutagenicity and micronucleus formation than the other vegetable juice.

In Vitro Screening of Antibacterial Agents for Suppression of Fire Blight Disease in Korea (기내 검정법을 이용한 국내 과수 화상병 방제제 선발)

  • Lee, Min Su;Lee, Ingyeong;Kim, Sam Kyu;Oh, Chang-Sik;Park, Duck Hwan
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.41-51
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    • 2018
  • Since fire blight disease on apple and pear was produced in Korea in 2015, there were no registered chemicals to control against this disease. Instead, several antibacterial chemicals that were registered for other bacterial diseases such as soft rot and bacterial spot have been authorized by Rural Development Administration (RDA). However, these chemicals are not tested efficacy for fire blight disease except damage by those treatments on apple and pear in Korea. Thus, we evaluated efficiency using in vitro and in planta assays of antibacterial chemicals such as antibiotics and copper compounds including kasugamycin, oxytetracycline, oxolinic acid and streptomycin, and copper hydroxide, copper sulfate, oxine copper and tribasic copper sulfate, respectively. We also tested two kinds of biological agents. As expected, significant antibacterial effect was observed in vitro test of both antibiotics and copper-based chemicals. In planta test based on disease severity including ooze and water-soaked formation on immature pears, bacterial populations on blooms, and blight lesion formation in artificially inoculated shoots, kasugamycin, oxytetracycline and streptomycin have been shown the most efficiency among tested antibiotics. Four copper-based chemicals tested in this study, control effects are little bit lower than agricultural antibiotics but they seem to be available to use in terms of winter season. Biocontrol agents were also shown possibility to treat in eco-friendly farms. In addition, there are no antibiotic resistance genes in Korean isolates against antibiotics, which were selected for suppression of fire blight in this study.

In Vitro AChE Inhibition Tests of Insecticides Using Electric eel and Housefly AChE (전기뱀장어 및 집파리 AChE를 이용한 살충제의 In Vitro AChE 저해 시험)

  • 이시혁;이준호;조광연
    • Korean journal of applied entomology
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    • v.31 no.2
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    • pp.122-132
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    • 1992
  • Experiments were conducted to establish an in vitro AChE inhibition test system to evaluate the potency of AChE inhibition of new chemical compounds. For a fixed time inhibition test, optimal inhibition (incubation) time to evaluate their AChE inhibition potency was 10 min. for AChE inhibitors such as DFP, DDVP, and paraoxon. The concentration of new chemical compounds with an ester group for evaluation of their inhibition potency was 10 $\mu$M under 10 min. preincubation conditions. However, the stepwise inhibition test with higher concentrations seemed to be needed for other chemical compounds. For a progressive inhibition test to calculate inhibition constants such as $K_d$, $K_3$ and $K_i$, extremely low $K_d(1.3\times10-^85.6\times10^{-7})$ and $K_3$(0. 21-0.27 $min^{-1}$) were observed under lagged preincubation time (0.8-13.3 min) and low in¬hibitor concentrations $(1\times10-^92\times10-^6M)$. However, this method seemed to be useful for comparison of AChE inhibition potency among inhibitors. Differences in inhibition potency among DFP, paraoxon, and KH501 were due to the differences in $K_d$, in other words, differences in affinities between inhibitors and AChEs. Therefore, AntiChE screening should consist of two steps. The first step is to evaluate the potency of AChE inhibition based on $I_50$ valuse obtained from fixed time inhibition tests. The second step is to study inhibition patterns and characteristics of chemical compounds selected in the first step.

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Screening of Indigenous Strains of Lactic Acid Bacteria for Development of a Probiotic for Poultry

  • Karimi Torshizi, M.A.;Rahimi, Sh.;Mojgani, N.;Esmaeilkhanian, S.;Grimes, J.L.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.10
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    • pp.1495-1500
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    • 2008
  • In an attempt to develop a probiotic formulation for poultry feed, a number of lactic acid bacteria (LAB) were isolated from chicken intestinal specimens and a series of in vitro experiments were performed to evaluate their efficacy as a potential probiotic candidate. A total of 650 LAB strains were isolated and screened for their antagonistic potential against each other. Among all the isolates only three isolates (TMU121, 094 and 457) demonstrated a wide spectrum of inhibition and were thus selected for detailed investigations. All three selected isolates were able to inhibit the growth of E. coli and Salmonella species, although to variable extent. The nature of the inhibitory substance produced by the isolates TMU121 and 094 appeared to be associated with bacteriocin, as their activity was completely lost after treatment with proteolytic enzymes, while pH neutralization and catalase enzyme had no effect on the residual activity. In contrast, isolate TMU457 was able to resist the effect of proteolytic enzymes while pH neutralization completely destroyed its activity. Attempts were made to study the acid, bile tolerance and cell surface hydrophobicity of these isolates. TMU121 showed high bile salt tolerance (0.3%) and high cell surface hydrophobicity compared to the other two strains studied, while TMU094 appeared the most pH resistant strain. Based on these results, the three selected LAB isolates were considered as potential ingredients for a chicken probiotic feed formulation and were identified to species level based on their carbohydrate fermentation pattern by using API 50CH test kits. The three strains were identified as Lactobacillus fermentum TMU121, Lactobacillus rhamnosus TMU094, and Pediococcus pentosaceous TMU457.

Glycosylation of Semi-Synthetic Isoflavene Phenoxodiol with a Recombinant Glycosyltransferase from Micromonospora echinospora ATCC 27932

  • Seo, Minsuk;Seol, Yurin;Park, Je Won
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.657-662
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    • 2022
  • Glycosyltransferase (GT)-specific degenerate PCR screening followed by in silico sequence analyses of the target clone was used to isolate a member of family1 GT-encoding genes from the established fosmid libraries of soil actinomycetes Micromonospora echinospora ATCC 27932. A recombinant MeUGT1 was heterologously expressed as a His-tagged protein in E. coli, and its enzymatic reaction with semi-synthetic phenoxodiol isoflavene (as a glycosyl acceptor) and uridine diphosphate-glucose (as a glycosyl donor) created two different glycol-attached products, thus revealing that MeUGT1 functions as an isoflavonoid glycosyltransferase with regional flexibility. Chromatographic separation of product glycosides followed by the instrumental analyses, clearly confirmed these previously unprecedented glycosides as phenoxodiol-4'-α-O-glucoside and phenoxodiol-7-α-O-glucoside, respectively. The antioxidant activities of the above glycosides are almost the same as that of parental phenoxodiol, whereas their anti-proliferative activities are all superior to that of cisplatin (the most common platinum chemotherapy drug) against two human carcinoma cells, ovarian SKOV-3 and prostate DU-145. In addition, they are more water-soluble than their parental aglycone, as well as remaining intractable to the simulated in vitro digestion test, hence demonstrating the pharmacological potential for the enhanced bio-accessibility of phenoxodiol glycosides. This is the first report on the microbial enzymatic biosynthesis of phenoxodiol glucosides.

Inhibitory Effect and Mechanism on Melanogenesis of Radix glycyrrhizae Water Extract (감초 물추출물의 멜라닌 형성 억제효과 및 기전에 관한 연구)

  • Kim, Jin;Kwen, Il-ho;Lim, Hong-jin;Lim, Kyu-sang;Hwang, Chung-yeon
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.16 no.2
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    • pp.96-118
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    • 2003
  • The effect of Glycyrrhizae Radix water extract, known as depigmenting agent, on melanin biosynthesis was investigated in cellular level by using B16 mouse melanoma cells. The inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis was determined by mushroom tyrosinase assay traditionally using in vitro screening test. To determine whether Glycyrrhizae Radix water extract suppress melanin synthesis in cellular level, B16 mouse melanoma cells were cultured in the presence of different concentrations of Glycyrrhizae Radix water extract. Effects on cell proliferation, melanin biosynthesis, tyrosinase activity, DOPAchrome tautomerase activity, and expression level of mRNA for tyrosinase were examined. The maximum concentration of Glycyrrhizae Radix water extract that was not inhibitory to growth of the cells was 2 mgml. At that concentration, melanin synthesis was significantly inhibited without cytotoxicity after 5 days, compared with untreated cells. The treatment with Glycyrrhizae Radix water extract reduced tyrosinase and DOPAchrome tautomerase activity in a dose-dependent manner. However, the treatment with Glycyrrhizae Radix water extract did not affect significantly mRNA levels for tyrosinase. These results suggest that the inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis is correlated with the suppression of tyrosinase and DOPAchrome tautomerase activity more than altering mRNA levels of tyrosinase.

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Screening of effective control agents against bacterial soft rot on Chinese cabbage in alpine area (고랭지 배추 무름병 방제를 위한 우수약제 선발)

  • Chung, Eun-Kyoung;Zhang, Xuan-Zhe;Yeoung, Young-Rog;Kim, Byung-Sup
    • The Korean Journal of Pesticide Science
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    • v.7 no.1
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    • pp.32-37
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    • 2003
  • Bacterial soft rot by Erwinia carotovora subsp. carotovora is one of the diseases causing the biggest problem in Chinese cabbage. Chemical screening was conducted to select effective agents for controlling bacterial soft rot. Control effect of antibiotics, plant activator, and Biokeeper (avirulent Erwinia) to soft rot were tested by in vitro assay, nursery test, and field experiment. The in vitro assay was done by paper disc method and potato slice method. The nursery test was performed by using mineral oil inoculation method with consistent disease induction. The in vitro assay showed that streptomycin, oxolinic acid, bronopol, and copper hydroxide significantly suppressed the growth of pathogenic bacterium and the decomposition of potato slice. However, plant activators including acibenzolar-S-methyl did not show the suppressive effect on the growth of pathogenic bacterium and the decomposition of potato slice. When applied by the nursery test condition using mineral oil inoculation method with Chinese cabbage 'Kangruckyeurum', Biokeeper, oxolinic acid, antibiotics streptomycin, validamycin, and copper compound provided 83.5%, 95.2%, 91.2%, 57.5% and 79.9% in control efficacy, respectively. However, the control effect of acibenzolar-S-methyl showed to be low to cause phytotoxicity. Also acibenzolar-S-methyl showed a significant control effect in the field experiment with Chinese cabbage 'Sanchon' in 2000, but the field experiment with Chinese cabbage 'Kangruckyeurum' in 2001 revealed it had phytotoxicity to Chinese cabbage. Such a difference was considered to be caused by differences in phytotoxic reaction of Chinese cabbage cultivars to the chemical. Streptomycin+copper, copper hydroxide and Biokeeper showed 79.7%, 71.9% and 60.9% in control efficacy, respectively, in the field experiment with Chinese cabbage 'Sanchon' in 2002.

Screening of Oriental Herbal Medicines for Antibacterial Activities

  • Bae, O-Sung;Hwang, Jae-Ock;Ahn, Duk-Kyun;Woo, Eun-Rhan;Seo, Seon-Hee;Kim, Hyoung-Ja;Park, Ho-Koon
    • Natural Product Sciences
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    • v.4 no.1
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    • pp.32-37
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    • 1998
  • The water extracts of oriental herbal medicines which have been clinically used to treat bacterial infections in Korea were screened for in vitro antibacterial activity by the paper disc assay method. Two Gram positive bacteria, Staphylococcus aureus SG511, Bacillus subtilis ATCC 6633 and two Gram negative bacteria, Escherichia coli 055, Pseudomonas aeruginosa 9027 were used as test organisms. Among 83 of the extracts tested, 25 were active against Staphylococcus aureus SG511, 9 were active against Bacillus subtilis ATCC 6633, while none showed inhibitory activity against Eschelichia coli 055 and Pseudomonas aeruginosa 9027. Among them, Hwangyonhaedoktang plus hwangyon, Chongwisan, and Ssangbaksan showed remarkably potent antibacterial activity.

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Quality and Availability Evaluation of Human Hepatocytes Isolated from Resected Partial Livers for Toxicology and Drug Metabolism Studies in Korea (독성 및 약물대사 연구를 위한 한국인 부분 간 유래 간세포의 품질 및 활용성 평가)

  • Noh, Jeong-Kwon;Jang, In Keun;Kim, Hyo Eun;Lee, Jong Eun;Yang, Mal Sook;Jang, Eun Mi;Lee, Ji-Hyun;Park, Hey-Jung;Kim, Young-A;Lee, Suk-Koo;Jeong, Ho-Sang;Ahn, Joon-Ik;Lee, Doo-Hoon
    • KSBB Journal
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    • v.29 no.1
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    • pp.58-66
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    • 2014
  • Demand for in vitro pharmacological evaluation and toxicity test using human hepatocytes has been increasing. In USA and Europe, human hepatocytes obtained from donated whole liver unsuitable for transplantation were distributed to researchers and deposited in cell bank facility as cryopreserved vial. In Korea, however, incidence of transplantation- inappropriate whole liver has been quite low and the whole livers almost have so severe liver disease such as fatty or fibrotic liver that cannot meet the demand. In this study we aimed to isolate human hepatocytes from liver resection surgery-originated partial liver, and assure the isolated human hepatocytes and its cryopreserved hepatocytes to be qualified for the in vitro pharmacological evaluation and drug toxicity tests. We compared those with commercially available human hepatocyte, BD $GenTest^{TM}$ by cell morphology, hepatic gene expression, urea synthesis, albumin secretion, ammonia removal, and cytochrome P450 induction activities. Changes in hepatotoxic gene expression after cryopreservation are evaluated with a typical hepatotoxic drug, acetaminophen. Consequently, the fresh hepatocytes from the partial liver and its cryopreserved hepatocytes expressed their intrinsic hepatic functions well and showed equal hepatotoxicity gene expression trend regardless to cryopreservation. Therefore, liver resection surgery-originated partial liver can be used as a useful source of human hepatocytes for various pharmacological and hepatotoxicity test.

KRG and its major ginsenosides do not show distinct steroidogenic activities examined by the OECD test guideline 440 and 456 assays

  • Namkyu Lee;Ju Hyeong Lee;Ji Eun Won;Youn Ji Lee;Sun Hee Hyun;Yeong-Deuk Yi ;Gyo In;Hee Dong Han;YoungJoo Lee
    • Journal of Ginseng Research
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    • v.47 no.3
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    • pp.385-389
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    • 2023
  • Background: Ginseng has been used as a traditional medicine for treatment of many diseases and for general health maintenance. Previously, we showed that ginseng did not demonstrate estrogenic property in ovariectomized mouse model. However, it is still possible that disruption of steroidogenesis leading to indirect hormonal activity. Methods: The hormonal activities were examined in compliance with OECD guidelines for detecting endocrine disrupting chemicals: test guideline (TG) No. 456 (an in vitro assay method for detecting steroidogenesis property) and TG No. 440 (an in vivo short-term screening method for chemicals with uterotrophic property). Results: Korean Red Ginseng (KRG) and ginsenosides Rb1, Rg1, and Rg3 did not interfere with estrogen and testosterone hormone synthesis as examined in H295 cells according to TG 456. KRG treatment to ovariectomized mice did not show a significant change in uterine weight. In addition, serum estrogen and testosterone levels were not change by KRG intake. Conclusion: These results clearly demonstrate that there is no steroidogenic activity associated with KRG and no disruption of the hypothalamic-pituitary-gonadal axis by KRG. Additional tests will be performed in pursuit of cellular molecular targets of ginseng to manifest mode of action.