• Title/Summary/Keyword: Chromosomal technology

검색결과 149건 처리시간 0.028초

고전압 펄스 전기장에 의한 Saccharomyces cerevisiae의 세포내·외적 사멸 기작 연구 (Intra- and Extra-cellular Mechanisms of Saccharomyces cerevisiae Inactivation by High Voltage Pulsed Electric Fields Treatment)

  • 이상재;신정규
    • 한국식품과학회지
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    • 제47권1호
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    • pp.87-94
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    • 2015
  • 비가열 살균 기술 중 본격적인 상업적 실용화를 눈앞에 두고 있는 고전압 펄스 전기장에 의한 미생물의 사멸 기작에 대해 살펴보았다. 세포 현탁액을 고전압 펄스 전기장 처리하였을 경우 처리 시간이나 전기장의 세기가 증가할수록 세포 외액으로 세포내 물질의 유출이 증가하였으며, 세포막 투과성의 변화로 인하여 $K^+$, $Na^+$등이 이온 성분의 유출도 나타났다. 염색시약에 의한 세포의 염색에서 처리시간이 증가함에 따라 염색되는 세포의 수가 증가하였으며, 전자현미경에 의한 세포의 관찰 결과 고전압 펄스 전기장 처리를 받은 세포의 경우 처리 받지 않은 것에 비해 표면이 거칠고 굴곡이 있었으며, 세포막이 터져 세포내 물질이 외부로 유출되고 형태가 일그러진 것이 관찰되었다. 항생물질 첨가에 따른 회복 실험에서 고전압 펄스 전기장 처리에 의해 세포의 단백질 합성 체계에 손상을 입었으며, chromosomal DNA의 분리를 통한 DNA의 손상여부 관찰 결과 약 27.3%의 DNA의 손상이 발생했음을 알 수 있었다. 따라서 고전압 펄스 전기장 처리가 세포벽이나 세포막의 손상뿐만 아니라 대사 체계와 DNA에도 손상을 주는 것을 확인할 수 있었다.

Genotoxicity Study of Dimethyl Isophthalate in Bacterial and Mammalian Cell System

  • Chung, Young-Shin;Choi, Seon-A;Hong, Eun-Kyung;Ryu, Jae-Chun;Lee, Eun-Jung;Choi, Kyung-Hee
    • Molecular & Cellular Toxicology
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    • 제3권1호
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    • pp.53-59
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    • 2007
  • This study was conducted to evaluate the mutagenic potential of dimethyl isophthalate (DMIP) using Ames bacterial reverse mutation test, chromosomal aberration test and mouse lymphoma $tk^{+/-}$ gene assay. As results, in Ames bacterial reversion assay, DMIP was tested up to the concentration of 5,000 ${\mu}g$/plate and did not induce mutagenicity in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537, and Escherichia coli WP2uvrA with or without metabolic activation (S9 mix). Using cytotoxicity test, the maximal doses of DMIP for chromosomal aberration assay were determined at 1,250 ${\mu}g/mL$, which was a minimum precipitation concentration ($IC_{50}>1,940\;{\mu}g/mL$ or 10 mM) and at 155 ${\mu}g/mL$ ($IC_{50}:155\;{\mu}g/mL$) in the presence and the absence, respectively, of S9 mix. DMIP in the presence of S9 mix induced statistically significant (P<0.001) increases in the number of cells with chromosome aberrations at the dose levels of over 250 ${\mu}g/mL$, when compared with the negative control. However, DMIP in the absence of S9 mix did not caused significant induction in chromosomal aberrant cells. In MLA, DMIP at the dose range of 242.5-1,940 ${\mu}g/mL$ in the presence of S9 mix induced statistically significant increases in mutation frequencies related to small colony growth, whereas any significant mutation frequency was not observed in absence of S9 mix. From these results, it is conclusively suggested that dimethyl isophthalate may be a clastogen rather than a point mutagen.

Genotoxicity Study of Water Extract of Anemarrhena asphodeloides and Phellodendron amurense in Bacterial and Mammalian Cell Systems

  • Chung, Young-Shin;Lee, Seok-Jong;Choi, Sun-A;Lee, Jang-Ha;Ryu, Jae-Chun;Hong, Eun-Kyung
    • Toxicological Research
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    • 제20권1호
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    • pp.43-47
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    • 2004
  • In order to investigate the safety of a water extract (ADP) of 1 : 1 mixture of Anemarrhena rhizoma and Phellodendron cortex for alleviating benign prostate hyperplasia, genotoxicity studies in bacterial and mammalian cell assay systems, namely, the Ames bacterial reverse mutation and chromosomal aberration assays were performed. As shown by the results of the Ames bacterial reversion assay, ADP in the range of 625-5000 $\mu\textrm{g}$/plate did not induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains in the absence or in the presence of S9 (the microsomal fraction of rat liver homogenate) metabolic activation. The $IC_{50}$ (50% cell growth inhibition concentration) values of ADP for the chromosomal aberration assay were determined; these were 2425 $\mu\textrm{g}$/ml in the absence and 8126 $\mu\textrm{g}$/ml in the presence of S9 metabolic activation in Chinese hamster lung (CHL) fibroblast cell culture. No chromosomal aberration was observed in CHL cells treated with ADP at 2425, 1212.5 and 606.25 $\mu\textrm{g}$/ml in the absence, or at 8126, 4063 and 2031.5 $\mu\textrm{g}$/ml in the presence of S9 metabolic activation. These results show that under the conditions used, ADP does not harmfully affect the bacterial or mammalian cell system at the gene level.

Comparison of Sexing Analysis between Karyotyping and Blasomere-PCR in Bovine embryos

  • Chang, Suk-Min;Lee, Jong-Ho;Park, Joong-Hoon;Park, Wha-Sik;Park, Chang-Sik;Jin, Dong-Il
    • 한국발생생물학회:학술대회논문집
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    • 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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    • pp.92-92
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    • 2003
  • Accurate analysis of nuclear status is needed when biopsied-blastomeres are used for embryo sexing. In this study, the nuclear status of blastomeres derived from 8- to 16-cell stage IVF bovine embryos was analyzed to evaluate the representative of single blastomere for embryo sexing. When 55 embryos were analyzed by PCR following biopsy, the coincident rate of sex determination between biopsied-single blastomere and matched blastocyst by PCR was 80 %. Karyotyping of biastomeres in 8- 16-cell stage bovine embryos was conducted to assess chromosome status of IVF embryos. To establish karyotyping of blastomeres, concentrations of vinblastine sulfate and duration of exposure time for metaphase plate induction with 8- to 16-cell stage bovine embryos were tested. The most effective condition for induction of metaphase plate (>45%) was 1.0 ug/ml vinblastine sulfate treatment for 15 h. In 22 embryos under the condition, only 8 embryos out of ten that had a normal diploid chromosome complement showed a sex-chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four out of the other 11 embryos having a mixoploid chromosomal complement contained haploid blastomere with wrong sex chromosome (18.2%). These results suggested that morphologically normal bovine embryos derived from IVF had considerable proportion of mixoploid and sex-chromosomal mosaicism which could be the cause of discrepancies of the sex between biopsied-single blastomere and matched blastocyst by PCR analysis.

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Evaluation of the Genetic Toxicity of Synthetic Chemicals [XII] -in vitro Chromosomal Aberration Assay with 11 Chemicals in Chinese Hamster Lung Fibroblast-

  • Ryu, Jae-Chun;Kim, Youn-Jung
    • 한국환경성돌연변이발암원학회지
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    • 제24권2호
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    • pp.99-107
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    • 2004
  • The validation of many synthetic chemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, the regulation and evaluation of the chemical hazard playa very important role to environment and human health. The clastogenicity of 11 synthetic chemicals was evaluated in Chinese hamster lung (CHL) fibroblast in vitro. Benzoyl chloride (CAS No. 98-88-4) induced chromosomal aberrations with statistical significance at the concentration of 31-123 $\mug/ml$ and 43 $\mug/ml$ in the absence and presence of S-9 metabolic activation system, respectively. 2-Propyn-l-o1 (CAS No. 107-19-7) and 2-Phenoxy ethanol (CAS No. 122-99-6) revealed clastogenicity only at the highest concentration in the presence of S-9 mixture. However, 1-naphthol (CAS No. 90-15-3) which is one of the most cytotoxic chemical among 11 chemicals tested revealed no clastogenicity both in the presence and absence of S-9 metabolic activation system. From the results of chromosomal aberration assay with 11 synthetic chemicals in CHL fibroblast in vitro, Benzoyl chloride (CAS No. 98-88-4), 2-Propyn-l-01 (CAS No. 107-19-7) and 2-Phenoxy ethanol (CAS No. 122-99-6) revealed positive clastogenic results in this study.

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Congenital Malformations in Infants of Mothers Undergoing Assisted Reproductive Technologies: A Systematic Review and Meta-analysis Study

  • Hoorsan, Hayedeh;Mirmiran, Parvin;Chaichian, Shahla;Moradi, Yousef;Hoorsan, Roza;Jesmi, Fatemeh
    • Journal of Preventive Medicine and Public Health
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    • 제50권6호
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    • pp.347-360
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    • 2017
  • Objectives: This meta-analysis aimed to evaluate congenital malformations in infants conceived by assisted reproductive techniques (ART), compared with infants conceived spontaneously. Methods: In this study, available resources searched to find relevant articles included PubMed, ScienceDirect, Scopus, Google Scholar, Cochrane, ProQuest, Iranmedex, Magiran, and Scientific Information Database. After extracting the necessary information from evaluated articles, meta-analysis on the articles' data was performed using Stata version 11.2. Results: In this study, from a total of 339 articles, extracted from the initial investigation, ultimately 30 articles were selected for meta-analysis that assessed the use of ART on the risk of congenital abnormalities and some birth complications on 5 470 181 infants (315 402 cases and 5 154 779 controls). The odds ratio (95% confidence interval [CI]) for low birth weight was 1.89 (95% CI, 1.36 to 2.62), preterm labor 1.79 (95% CI, 1.21 to 2.63), cardiac abnormalities 1.43 (95% CI, 1.27 to 1.62), central nervous system abnormalities 1.36 (95% CI, 1.10 to 1.70), urogenital system abnormalities 1.58 (95% CI, 1.28 to 1.94), musculoskeletal disorders 1.35 (95% CI, 1.12 to 1.64), and chromosomal abnormalities in infants conceived by ART was 1.14 (95% CI, 0.90 to 1.44), which were all statistically significant, except chromosomal abnormalities. Conclusions: The risk of congenital abnormalities and some birth complications were significantly higher in ART than normal conception, while chromosomal abnormalities were not; therefore, the application of ART should be selected individually for patients by detailed assessment to reduce such risks in the population.

유기인화합물 분해효소 유전자의 재조합 및 단백질 발현 (Subcloning and Expression of a Gene Encoding an Organophosphorus Acid Anhydrolase)

  • 박재왕;김석찬;이남택
    • 한국군사과학기술학회지
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    • 제4권1호
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    • pp.188-197
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    • 2001
  • Organophosphorus acid anhydrolases(OPAA) catalyzing the hydrolysis of toxic organophosphates have been found in a variety of prokaryotic and eukaryotic organisms. Of the several kinds of OPAA that can degrade nerve agents, such as DFP, sarin and soman, a OPAA gene harbored in the chromosomal DNA of Alteromonas haloplanktis strain was subcloned in order to develope an enzymatic degradation method of toxic organophosphorus compounds. For this 1481 bp DNA fragment containing OPAA gene and its flanking regions has been synthesized through PCR using chromosomal DNA of A. haloplanktis strain. After subcloning and subsequent expression, crude OPA anhydrolase was prepared and assayed. It was shown that the OPAA had a very high hydrolytic activity on DFP. The specific activity of the enzyme was 1,110 $\mu$mole.$min^{p-1}.mg^{-1}$ protein. It seemed that OPAA with such a high hydrolytic activity may give a good prospects to its use, as a biodegradation tool, in detoxifying toxic organophosphorus compounds, such as pesticides and chemical stockpiles which are posing a potential threat to the field environment and human health.

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Comparison of L5178Y tk+/- Mouse Lymphoma Assay and In vitro Chromosome Aberration Test

  • Lee, Michael;Jung Kwon;Cho, Ji-Hee;Hong, Mi-Young;Kim, Eun-Joo;Junghee Han;Chung, Moon-Koo;Han, Sang-Seop
    • Toxicological Research
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    • 제18권2호
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    • pp.129-138
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    • 2002
  • The mouse lymphoma assay (MLA) has been recently validated as a sensitive and specific test system to determine the genotoxic potential for a chemical. The objective of this study is to evaluate the utility of MLA for detecting mutagens. Especially, to compare MLA with the in vitro chromosomal aberration test (CA), we performed MLA using the microwell method with three chemicals (hydroxyurea, theophylline and amino acid copper complex), which were reportedly positive in the CA. In cell treated with hydroxyurea, anti-neoplastic agent that blocks DNA replication, evidence of a positive response was obtained without S9 mix for 4 h and 24 h. In addition, analysis of colony size distribution at concentration that gave an elevated mutant fraction showed that hydroxyurea induced a high proportion of small type colonies, indicating that hydroxyurea-induced mutation is associated with large chromosomal deletion. Conversely, negative MLA result was obtained for theophylline, which was wed as central nervous system stimulator. Although theophylline increased the mutant frequency at concentration of 1250 $\mu\textrm{g}$/$\textrm{m}{\ell}$ with S9 mix for 4 h, a concentration-related increase in mutant frequency was not observed. The MLA result of amino acid copper complex was considered equivocal because the positive result was obtained at concentration showing 10% or less RS or RTG. Thus, among 3 CA-positive chemicals, positive MLA result was obtained for one. The other two chemicals were negative and equivocal. However MLA, which evaluates mutagenic potential of chemicals through colony formation by cell grouth, may provide a higher predictivity of carcinogenesis than CA.

Evaluation of the Genetic Toxicity of Synthetic Chemicals (IV) - Chromosomal Aberration Assay With 11 compounds in Chinese Hamster Lung Cells in vitro -

  • Kim, Kyung-Ran;Kim, Hyun-Joo;Kim, Min-Hee;Youn, Ji-Youn;Kwon, Oh-Seung;Ryu, Jea-Chun
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 1996년도 제19회정기학술대회(The 19th Symposium of the Korean Society of Environmental Toxicology)
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    • pp.63-63
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    • 1996
  • We performed chromosomal aberration assay in Chinese Hamster Lung (CHL) cells in vitro to evaluate theclastogenicity of 11 synthetic chemicals which were listed in Toxicity Evaluation Program of Ministry of Environment of Republic of Korea in 1996. All of the chemicals were carried out MTT assay to determine the 50% cell growth inhibition concentration. All compounds were tested with and without metabolic activation system. Benzoyl chloride revealed positive result at $43\;\mu\textrm{g}/m{\ell}$ in the presence of metabolic activation system, and at 30.8, 61.5 and $123\;\mu\textrm{g}/m{\ell}$ in the absence of metabolic activation system. And p-phenoxy ethanol was observed as positive with the metabolic activation system, but negative without metabolic activation system. Especially 2-propyn-l-ol showed high frequency of pulverization and showed critical difference of cytotoxicity between with and without S9 mixture. Pulverizatiuon is not included in the frequency of structural aberration in our criteria. Dicyclopentadiene, methacrylic acid, aa-dimethylbenzyl hydroperoxide, benzylbutyl phthalate, and p-chlorophenal were revealed negative results.esults.

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