• 제목/요약/키워드: in vitro detection

검색결과 263건 처리시간 0.022초

위암 조직의 자기공명영상과 초음파 소견에 대한 비교 연구 (In Vitro Imaging of MRI and Ultrasound for Gastric Carcinoma)

  • 길성원;지금난
    • Investigative Magnetic Resonance Imaging
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    • 제12권2호
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    • pp.178-187
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    • 2008
  • 목적 : 위암 조직의 종양 침범 깊이와 주위 조직으로의 침윤 정도를 초음파와 자기공명영상을 시행하여 그 진단적 유용성을 알아보고 자기공명영상의 경우 종양을 가장 잘 보여주는 펄스 연쇄에 대해 알아보고자 하였다. 대상 및 방법 : 위의 부분 혹은 전절제술을 받아 병리적으로 위암으로 확진된 53예의 제거된 위암 조직을 대상으로 하였다. 모든 조직은 자기공명영상과 고주파수 탐촉자로 초음파 영상을 얻었다. 각 조직에 대하여 자기공명영상과 초음파 소견을 각각 독립적으로 종양의 탐지 및 종양의 침범 깊이에 대하여 두 명의 방사선과 전문의가 합의하에 평가하였고 각각의 영상 소견을 병리 조직 소견과 비교하여 두 영상 기기 간에 진단의 정확도를 비교하였다. 자기공명영상은 스핀에코 T1 강조 영상, 위상 및 탈위상 경사에코 T1 강조영상, 고속스핀에코 및 단발포고속스핀에코 T2 강조영상의 다섯 펄스 연쇄를 얻었고 이 중 종양의 탐지와 묘출에 우수하다고 평가된 영상 기법을 알아보았다. 결과 : 조기 위암의 경우, 초음파 진단의 정확도는 77%로 자기공명영상의 진단적 정확도 59% 보다 우수하였으나 통계적으로 유의한 차이는 없었다 (p=0.096). 진행성 위암의 경우 자기공명영상이나 초음파 각각 97%와 84%의 높은 진단적 정확도를 보여주었으며 자기공명영상이 초음파에 비하여 통계적으로 유의하게 종양병기를 정확하게 진단했다 (p<0.001). 자기공명영상의 다섯 가지 펄스연쇄 중에 종양 침윤 깊이를 선명하고 정확하게 보여준다고 평가된 영상은 조기 위암과 진행성 위암 모두 고속스핀에코 T2 강조영상(75%)이었고, 특히 진행성 위암의 경우 총 93.5%에서 고속스핀에코 T2 강조영상이 우수한 영상 소견을 보였다. 결론 : 자기공명영상과 초음파는 진행성 위암 조직의 종양 병기를 평가하는데 높은 진단적 정확성을 갖고 있으며 자기공명영상이 초음파보다 통계적으로 유의하게 종양병기를 정확하게 진단했다. 위암 조직의 종양 병기를 평가하는데 가장 우수한 자기공명영상 기법은 고속스핀에코 T2 강조영상이었다.

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Adenosine Deaminase 표지유전자로 형질전환된 연초의 신속한 Assay 방법 (Visible and Fast Assay System for Tobacco Transformant Introduced with Adenosine Deaminase Marker Gene)

  • 양덕춘;김용환;임학태;방극수;배창휴
    • 식물조직배양학회지
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    • 제28권3호
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    • pp.165-171
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    • 2001
  • Adenosine deaminase 유전자를 연초의 형질전환용 표지유전자로 활용할 때 형질전환체 여부를 매우 빠르고 눈으로 직접 색깔을 확인할 수 있는 새로운 방법이 개발되었다. ADA 효소는 독성인 adenosine 유도체를 비독성인 inosine 유도체와 암모니아로 변환시키는데, 이때 형성된 암모니아를 phenol-nitoprusside와 alkaline-hypochlorite 용액을 이용하여 청색으로 변환시켜 96 well plate상에서 1시간 내에 형질전환체 여부를 쉽게 확인할 수 있게 되었다. ADA효소의 substrate로서 9-D-arabinofuranosyl adenine, cordycepin, 2'-deoxyadenosine, adenosine and xylofuranosyl adenine이 모두 가능하였으며, substrate 용액의 최적조건은 adenosine 10 mM과 pH 7.5이었다. 특히 형질전환체는 ADA효소의 inhibitor인 deoxycoformycin이 함유되어 있는 용액 속에서는 adenosine을 inosine과 암모니아로 변환시키지 못해 색깔의 변화가 없었는데, 이는 형질전환체에서 색깔의 변화는 ADA효소의 작용 때문에 일어나는 것을 의미한다. 따라서 본 연구결과는 ADA 표지유전자가 도입된 형질전환체의 확인에 있어서 GUS gene system과 같이 눈으로 직접 확인할 수 있을 뿐만 아니라 매우 작은 크기의 형질전환체 절편으로 쉽고, 빠르면, 값싸게 확인할 수 있게 되었다.

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누에를 이용한 곤충병원성 선충의 검색 및 배양 (The Detection and Multiplicity of Entomopathogenic Nematodes Using Silkworms (Bombyx mori))

  • 한상미;이광길;여주홍;권해용;우순옥;이용우;백하주;한명세
    • 한국잠사곤충학회지
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    • 제45권2호
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    • pp.103-107
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    • 2003
  • 본 연구는 해충에 대한 생물적 방제제로서 유용한 곤충병원성 선충의 효율적 검색과 in vitro에서의 대량 증식 가능성을 조사하였으며 다음과 같은 결과를 얻었다. 1. 토양으로부터 곤충병원성 선충의 검색을 위한 trap용 곤충으로는 꿀벌부채명나방보다 누에가 감수성이 높았다. 2. Sterinernema, Rhabditidae, Diplogastroidae과 계통의 선충은 꿀벌부채명나방 유충에서의 증식은 5만 마리 이하였으나, 누에 유충에서는 15만에서 35만 마리의 선충 증식이 일어났으며, 누에 번데기에서는 25만 마리 이상의 선충이 증식되었다. 누에 증식에 있어 누에 번데기를 사용했을 때가 가장 우수하였다. 3. Sterinernema, Rhabditidae, Diplogastroidae과 계통의 선충에 대한 증식률은 열풍건조 번데기의 흡수처리 후 배양 및 기타 천연물 인공배지에서 증식은 가능하였으나 살아있는 누에 유충보다는 증식률이 낮았다. 4. 이상의 결과로 토양으로부터 곤충병원성 선충을 분리하는데 누에를 이용한 trap이 매우 우수하였으며, 선충의 증식에 있어 누에 유충 및 번데기가 효과적인 것으로 판단된다.

Di(2-ethylhexyl) phthalate에 의해 유도된 DNA손상과 소핵 형성 (DNA Damage and Micronuclei Induced by Di (2-ethylhexyl) phthalate in Human Breast Carcinoma MCF-7 cells)

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • 한국환경성돌연변이발암원학회지
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    • 제21권1호
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    • pp.34-43
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    • 2001
  • Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

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Localization of Autophagosome in Porcine Follicular Cumulus-oocyte Complex

  • Lee, Seunghoon;Kim, Dong-Hoon;Im, Gi-Sun;Ock, Sun-A;Ullah, Imran;Hur, Tai-Young
    • 한국수정란이식학회지
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    • 제32권3호
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    • pp.105-109
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    • 2017
  • Autophagy is an intracellular degradation and recycling system. Oocyte maturation is dynamic process, in which various proteins should be synthesized and degraded. In our previous study, we reported the loci of autophagosome and dynamics of autophagic activity in porcine oocytes during in vitro maturation. In this study, we verified loci of autophagosome in porcine follicular cumulus-oocyte complex by detection of microtubule-associated protein 1A/1B-light chain 3 (LC3) which is the reliable marker of autophagosome. Porcine ovary including various sizes of follicles was fixed within 1 hour after collection from slaughterhouse. After fixation, immunohistochemistry was conducted on sliced ovary tissue containing various sizes of follicles by using LC3 antibody. As a result, LC3 signal was clearly detected in both cumulus and oocytes of various sizes of follicles. We also found ring shaped signal which represent autophagosome near oocyte membrane. Most of the signals in oocytes were localized nearby cellular membrane while evenly dispersed in cumulus cells. Therefore, this result suggests that autophagy occurs in porcine COCs (cumulus-oocyte complexes) at follicular stage.

Curcumin Induces Caspase Mediated Apoptosis in JURKAT Cells by Disrupting the Redox Balance

  • Gopal, Priya Kalyan;Paul, Mausumi;Paul, Santanu
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권1호
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    • pp.93-100
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    • 2014
  • Background: Curcumin has has been reported to exert anti-inflammatory, anti-oxidation and anti-angiogenic activity in various types of cancer. It has also been shown to induce apoptosis in leukemia cells. We aimed to unravel the role of the redox pathway in Curcumin mediated apoptosis with a panel of human leukemic cells. Materials and Methods: In this study in vitro cytotoxicity of Curcumin was measured by MTT assay and apoptotic effects were assessed by annexin V/PI, DAPI staining, cell cycle analysis, measurement of caspase activity and PARP cleavage. Effects of Curcumin on intracellular redox balance were assessed using fluorescent probes like $H_2DCFDA$, JC1 and an ApoGSH Glutathione Detection Kit respectively. Results: Curcumin showed differential anti-proliferative and apoptotic effects on different human leukemic cell lines in contrast to minimal effects on normal cells. Curcumin induced apoptosis was associated with the generation of intracellular ROS, loss of mitochondrial membrane potential, intracellular GSH depletion, caspase activation. Conclusions: As Curcumin induces programmed cell death specifically in leukemic cells it holds a great promise as a future therapeutic agent in the treatment of leukemia.

Analysis of in vitro apoptosis induced by virulent Korean isolate of classical swine fever virus in peripheral blood B cell line

  • Kim, Seon-Mi;Lim, Seong-In;Song, Jae-Young;Hyun, Bang-Hun
    • 대한수의학회지
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    • 제52권4호
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    • pp.259-262
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    • 2012
  • Classical swine fever (CSF) is a highly contagious disease among swine that has an important economic impact on worldwide. One clinical symptom of CSF is leukopenia, in particular lymphopenia, which is a characteristic event that occurs early in the course of CSF. Though lymphopenia associated with apoptosis, the pathogenic mechanism underlying the lymphopenia has not been well studied. To understand these mechanisms, we investigated the response of porcine B cell lines to infection with SW03, virulent strain isolated from swine tissue in Korea. This study demonstrated that SW03-infected L35 cell were induced apoptosis through the detection of activated caspase-3. In addition, SW03 infection leaded to alterations in pro-apoptotic, Bax, and anti-apoptotic, Bcl-xL proteins of Bcl-2 family. Our results would suggest that SW03-infected L35 cells induced apoptosis via intrinsic mitochondrial pathway.

HBD: A new tool to enhance human skin self-defence against micro-organisms

  • Ingrid Pernet;Corinne Reymermier;Anne Guezennec;Jacqueline Viac;Branca, Jean-Eric;Joelle Guesnet;Eric Perrier
    • 대한화장품학회:학술대회논문집
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    • 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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    • pp.85-96
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    • 2003
  • Normal human skin, constantly challenged by environmental micro-organisms, has an innate ability to fight invading microbes through antimicrobial peptides. These peptides, described in both plant and animal kingdoms are able to inactivate a broad spectrum of micro-organisms. Mammalian defensins constitute one of the most common antimicrobial peptide family. Among the three human beta-defensins hBD1, hBD2 and hBD3 produced in epithelia, only hBD2 and hBD3 are inducible and additionally have been described as expressed by differentiated keratinocytes at site of inflammation and infection. The aims of these studies were to define a cell culture model in which the basal production of hBD could be detected and up-regulated in order to enhance skin auto-protection against micro-organisms. A specific Polymerase Chain Reaction method have been developed for hBD2 and hBD3 mRNA detection in non-differentiated monolayer keratinocytes cell culture. We have been able to demonstrate that in vitro, hBD2 and hBD3 expression in normal human keratinocytes could be detected and enhanced by TNF-alpha and IFN-gamma, in hypercalcic culture conditions. This research opened the possibility of the development of cosmetic active compounds, able to induce the expression of skin natural antibiotic peptides responsible about microflora ecology of the skin.

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환경 중 유전독성물질 검색을 위한 자주달개비 생물검정 기법의 적용연구 (Biomonitoring the Genotoxicity of Environmental Pollutants Using the Tradescantia Bioassay)

  • 신해식
    • 한국환경독성학회:학술대회논문집
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    • 한국환경독성학회 2004년도 춘계학술대회
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    • pp.47-60
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public from agents that can cause mutation and/or cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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자주달개비 생물검정 기법을 이용한 환경오염 평가 (Assessment of Environmental Pollution with Tradescantia Bioassays)

  • 김진규;신해식
    • 한국환경생물학회:학술대회논문집
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    • 한국환경생물학회 2004년도 학술대회
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    • pp.1-15
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public firom agents that can cause mutation anuor cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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