• 제목/요약/키워드: mammalian cell toxicity

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유전자 재조합 형광 단백질 발현 동물세포의 고정화 및 바이오센서의 개발

  • 이정은;구만복
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2002년도 생물공학의 동향 (X)
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    • pp.53-56
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    • 2002
  • Mammalian cell based biosensor kits are expected to be in assessment of samples toxicity more sensitive and accurate. A recombinant fluorescent Chinese Hamster Ovary (CHO) cell line was known to be responsive to the various toxicants Specially. KFC- AlO cell line. which contain the c-fos SRE::GFP plasmid (pKFG). was found to be able to detect toxicants sensitively. A biosensor kit was developed by using an immobilized KFC-A10 cell line. Immobilized recombinant fluorescent cells within agarose, known as a representative hydrogel matrix, have been maintained in the matrix viably and have shown constant fluorescent levels for long time. Immobilized cells have shown the ability to detect the chemical toxicity in the keep of fluorescent level as the metabolism is inhibited under toxic conditions.

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유산균 발효 쌍화탕에 대한 단회 투여 경구 독성 및 유전 독성 연구 (Single Dose Oral Toxicity and Genotoxicological Safety Study of Ssanghwa-tang Fermented with Lactobacillus acidophyllus)

  • 정태호;심기석;김동선;이재훈;마진열
    • 대한한의학회지
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    • 제32권1호
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    • pp.67-83
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    • 2011
  • Objectives: The purpose of this study was to examine the single dose toxicity with oral administration and genotoxicities of Ssanghwa-tang fermented with Lactobacillus acidophyllus. Materials and Methods: Clinical signs, weight changes, lethal doses$(LD_{50})$, and postmortem evaluation were determined by Globally Harmonized Classification System(GHCS) in a single-dose oral toxicity study. In vitro mammalian chromosomal aberration test was conducted with Ames test by cell proliferation suppression assessment using the cultivated CHO-K1(Chinese hamster ovary fibroblast) origins. Bacterial reversion assay was performed using Salmonella typhimurium (TA98, TA100, TA1535, and TA1537) and Escherichia coli (WP2uvrA). In vivo micronucleus test was performed using ICR mouse bone marrow. Results: No clinical sign was observed and none of the groups with doses up to 2000 mg/kg showed significant acute oral toxicity in the single dose oral administration. None of the sample doses taken during the 6 to 18 hour groups showed significant aberrant metaphases comparing to the negative control group in the in vitro mammalian chromosomal aberration test. No evidence of mutagenicity was seen for Escherichia coli (WP2uvrA) or Salmonella typhimurium (TA98, TA100, TA1535, and TA1537). No significant increase in the frequency of micronuclei was seen in the micronucleus test. Conclusion: These results indicate that the $LD_{50}$ value of Ssanghwa-Tang fermented with Lactobacillus acidophyllus may be over 2000 mg/kg and it have no acute oral toxicity and genotoxicity.

Application of Toxicogenomic Analysis to the Monitoring of Environmental Toxicity Using Recombinant Bioluminescent Bacteria and Cultured Mammalian Cells

  • Choi, Sue Hyung;Gu, Man Bock;Yasuyuki, Sakai
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2003년도 2003 Annual Meeting, BioExhibition and International Symposium
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    • pp.129-131
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    • 2003
  • Recombinant bioluminescent bacteria and cultured human cells were applied for toxicogenomic analysis of environmentally hazardous chemicals. Recombinant bioluminescent biosensing cells were used to detect and classify the toxicity caused by various chemicals. Classification of toxicity was realized based upon the chemicals' mode of action using DNA-, oxidative-, protein, and membrane-damage sensitive strains. As well, a simple double-layered cell culture system using Caco-2 cells and Hep G2 cells, which mimic the metabolic processes occurring in humans, such as adsorption through the small intestine and biotransformationin both the small intestine and the liver, was developed to investigate the toxicity of hazardous materials to humans. For a more in-depth analysis, a DNA microarray was used to study the transcriptional responses of Caco-2 and Hep G2 cells to benzo〔a〕pyrene.

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Evaluation of the Genetic Toxicity of Synthetic Chemicals (II), a Pyrethroid Insecticide, Fenpropathrin

  • Ryu, Jae-Chun;Kim, Kyung-Ran;Kim, Hyun-Joo;Ryu, Eun-Kyoung;Lee, Soo-Young;Jung, Sang-Oun;Youn, Ji-Youn;Kim, Min-Hee;Kwon, Oh-Seung
    • Archives of Pharmacal Research
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    • 제19권4호
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    • pp.251-257
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    • 1996
  • The detection of many synthetic chemicals used in industry that may pose a genetic hazard in our environment is subject of great concern at present. In this respect, the genetic toxicity of fenpropathrin ((RS)-.alpha.-cyano-3-phenoxybenzyl-2,2,3,3-tetramethyl cyclopropane carboxylate, CAS No.:39514-41-8), a pyrethroid insecticide, was evaluated in bacterial gene mutation system, chromosome aberration in mammalian cell system and in vivo micronucleus assay with rodents. In bacterial gene mutation assay, no mutagenicity of fenpropathrin (62-$5000\mug/plate$) was observed in Salmonella typhimurium TA 98, 100, 1535 and 1537 both in the absence and in the presence of S-9 metabolic activaton system. In mammalian cell system using chinese hamster lung fibroblast, no clastogenicity of fenpropathrin was also observed both in the absence and in the presence of metabolic activation system in the concentration range of $7-28\mug/ml$. And also, in vivo micronucleus assay using mouse bone marrow cells, fenpropathrin also revealed no mutagenic potential in the dose range of 27-105 mg/kg body weight of fenpropathrin (i.p.). Consequently, no mutagenic potential of fenpropathrin was observed in vitro bacterial, mammalian mutagenicity systems and in vivo micronucleus assay in the dose ranges used in this experiment.

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Fusarium mangiferae as New Cell Factories for Producing Silver Nanoparticles

  • Hamzah, Haider M.;Salah, Reyam F.;Maroof, Mohammed N.
    • Journal of Microbiology and Biotechnology
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    • 제28권10호
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    • pp.1654-1663
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    • 2018
  • Finding a safe and broad-spectrum medication is a goal of scientists, pharmacists, and physicians, but developing and fabricating the right medicine can be challenging. The current study describes the formation of silver nanoparticles (AgNPs) by Fusarium mangiferae. It involves the antibiofilm activity of the nanoparticles against Staphylococcus aureus. It also involves cytotoxic effect against mammalian cell lines. Well-dispersed nanoparticles are formed by F. mangiferae. The sizes of the nanoparticles were found to range from 25 to 52 nm, and UV-Vis scan showed absorption around 416-420 nm. SEM, TEM, and AFM results displayed spherical and oval shapes. Furthermore, the FTIR histogram detected amide I and amide II compounds responsible for the stability of AgNPs in an aqueous solution. AgNPs were observed to decrease the formation of biofilm at 75% (v/v). DNA reducing, smearing, and perhaps fragmentation were noticed after treating the bacterial cells with 50% (v/v). Additionally, cell lysis was detected releasing proteins in the supernatant. It was also observed that the AgNPs have the ability to cause 59% cervical cancer cell line (HeLa) deaths at 25% (v/v), however, they showed about 31% toxicity against rat embryo fibroblast transformed cell lines (REF). The results of this study prove the efficiency of AgNPs as an antibiofilm against S. aureus, suggesting that AgNPs could be an alternative to antibiotics. It must also be emphasized that AgNPs displayed cytotoxic behavior against mammalian cell lines. Further studies are needed for assessing risk in relation to the possible benefit of prescribing AgNPs.

고분자필름과 금속막 의료소재에 대한 생체적합성 및 독성 평가를 위한 새로운 세포배양시스템의 개발 및 적용 (Development and Application of a Novel Mammalian Cell Culture System for the Biocompatibility and Toxicity of Polymer Films and Metal Plate Biomaterials)

  • 곽문화;윤우빈;김지은;성지은;이현아;서은지;남국일;정영진;황대연
    • 생명과학회지
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    • 제26권6호
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    • pp.633-639
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    • 2016
  • 고분자(polymer), 금속(metal), 세라믹(ceramic), 합성물(composite) 등과 같은 바이오소소재(Biomaterials)는 그들의 물리화학적 성질 때문에 의료용섬유(medical fibers), 인공혈관(artificial blood vessels), 인공관절(artificial joints), 임플란트(implants), 연조직(soft tissue), 인공성형물(plastic surgery materials) 등 의료용으로 많이 사용되며, 개발연구도 활발히 진행되고 있다. 그러나, 필름(film)이나 판(plate)형태의 바이오소재에 대한 생체적합성(biocompatibility)이나 독성(toxicity)을 포유동물세포를 이용하여 평가하는 것은 적절한 평가용 장치가 없기 때문에 매우 어려운 상황이다. 따라서 이러한 문제를 해결하기 위해서, 본 연구에서는 고분자필름이나 금속판에 유용하게 적용할 수 있는 실리콘링, 상판(top panel), 하판(bottom panel)으로 구성된 새로운 포유동물배양시스템을 개발하고, 이를 실제 적용하고자 하였다. 개발된 시스템은 평가하고자 하는 시료를 상판과 하판사이에 조립하는 샌드위치시스템을 기반으로 한다. 세포배양장치의 조립 후, SK-MEL-2세포를 3가지 시료; Styela Clava Tunic (SCT)- PF, NaHCO3-added SCT (SCTN)-PF, magnesium MP (MMP)에 적용하고 37℃ 이산화탄소 배양기에서 24시간과 48시간 동안 배양하였다. MTT분석결과에서, 세포생존율(cell viability)은 24시간과 48시간 동안 SCT-PF배양그룹에서 정상적으로 유지되었지만 48시간 동안 SCTN-PF배양그룹에서는 급격하게 감소되었다. 더불어, MMP배양그룹에서 세포생존율은 24시간과 48시간 배양 후에 대조군과 유사하게 유지되었다. 이러한 결과는 본 연구에서 새롭게 개발된 샌드위치형태의 포유동물세포배양장치는 고분자필름이나 금속판형태의 바이오소재에 대한 독성이나 생체적합성을 평가하기 위한 우수한 잠재력을 보유하고 있음을 제시하고 있다.

A Novel Helicosporium Isolate and Its Antimicrobial and Cytotoxic Pigment

  • Choi, Hye Jung;Lee, Sang Myeong;Kim, Sun-Hee;Kim, Dong Wan;Choi, Young Whan;Joo, Woo Hong
    • Journal of Microbiology and Biotechnology
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    • 제22권9호
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    • pp.1214-1217
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    • 2012
  • One Helicosporium strain, isolated from a wilted chestnut tree, evidenced in vitro antimicrobial activity against various types of bacteria and fungi, and generated a diffusible pigment. The antimicrobial compounds and the diffusible pigment of the Helicosporium sp. isolate were purified via solvent fractionation, column chromatography, and recycling preparative chromatography. Both the major antimicrobial compound and the diffusible pigment were identified as 2-methylresorcinol via nuclear magnetic resonance spectroscopy. Therefore, 2-methylresorcinol, a diffusible pigment generated by Helicosporium sp., appears to be an active antimicrobial principle. This pigment also exhibited considerable cytotoxicity against mammalian cells.

Genotoxicity Study of sophoricoside derivatives in mammalian cells system

  • Yun, Hye-Jung;Kim, Youn-Jung;Kim, Eun-Young;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.178.2-178.2
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    • 2003
  • To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of Quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay(MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. (omitted)

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증가된 원핵세포선택성을 가진 짧은 인돌리시딘 유사체의 설계 (Design of Short Indolicidin Analogs with Enhanced Prokaryotic Selectivity)

  • 신송엽
    • 한국미생물·생명공학회지
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    • 제40권4호
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    • pp.409-413
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    • 2012
  • 인돌리시딘(indolicidin)은 소의 호중구(bovine neutrophils)로 부터 분리된 13개의 아미노산 잔기로 이루어 지고, 트립토판(tryptophan)을 많이 함유한 항균 펩타이드(antimicrobial peptide)이다. 인돌리시딘은 강력한 항균활성과 더불어 mammalian cells에 대해 독성을 나타낸다. 본 연구에서는 인돌리시딘 보다 서열이 짧으며, 보다 증가된 원핵세포 선택성(박테리아세포에 대해 독성을 나타내지만 mammalian cells에 대해서는 독성을 나타내지 않음을 의미함)을 지닌 새로운 짧은 항균 펩타아드를 개발하기 위해, 몇 종의 인돌리시딘 유사체 펩타이드를 설계하고 합성하였다. 결과적으로, 인돌리시딘 보다 서열이 짧으며(10개의 아미노산 잔기로 이루어짐), 증가된 원핵세포선택성을 지닌 4종의 새로운 펩타이드(SI, SI-PA, SI-WF 및 SI-WL)를 개발하였다. 본 연구를 통하여 가장 높은 원핵세포선택성을 나타내는 인돌리시딘 유사체 펩타이드 SI의 항균활성에는 중앙부위에 위치하는 소수성 및 방향족 아미노산이 중요하며, 2군데의 프로린(proline) 잔기는 중요하지 않다는 것을 알았다. 인돌리시딘과 유사체 펩타이드에 대한 원핵세포 선택성은 SI>SI-PA>SI-WF>SI-WL>ID>SI-WA의 순서 이였다. 따라서, 본 연구에서 설계되고 합성한 4종의 인돌리시딘 유사체 펩타이드(SI, SI-PA, SI-WF 및 SI-WL)는 박테리아 감염의 치료제로서 개발될 수 있을 것이다.

Genotoxicity Study on Khal, a Halocidin Derivative, in Bacterial and Mammalian Cells

  • Kim, Youn-Jung;Kim, Mi-Soon;Jeon, Hee-Kyoung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제2권3호
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    • pp.151-158
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    • 2006
  • Khal was a synthetic congener of halocidin, a heterodimeric peptide consisting of 19 and 15 amino acid residues detected in Halocynthia aurantium. This compound was considered a candidate for the development of a novel peptide antibiotic. The genotoxicity of Khal was subjected to high throughput toxicity screening (HTTS) because they revealed strong antibacterial effects. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay and chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of Khal was determined the concentration of $25.51\;{\mu}/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, Khal was not induced DNA damage in mouse lymphoma cell line. Also, the mutation frequencies in the Khal-treated cultures were similar to the vehicle controls. It is suggests that Khal is non-mutagenic in MOLY assay. And no clastogenicity was observed in Khal-treated Chinese hamster lung cells. The results of this battery of assays indicate that Khal has no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that Khal, as the optimal candidates with both no genotoxic potential and antibacterial effects must be chosen.