• Title/Summary/Keyword: in vitro detection

Search Result 263, Processing Time 0.025 seconds

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

  • Kil, Sung-Won;Jee, Keum-Nahn
    • Investigative Magnetic Resonance Imaging
    • /
    • v.12 no.2
    • /
    • pp.178-187
    • /
    • 2008
  • Purpose : To evaluate and compare the diagnostic accuracy of MRI and ultrasound(US) for estimation of invasion depth of gastric carcinoma by correlation with histopathologic findings in vitro and to find out the best MR pulse sequence for detection and accurate delineation of tumor. Materials and Methods : Resected specimen of total or subtotal gastrectomy from 53 patients with gastric carcinoma were done of imaging studies of MRI and US. And US was examined by using high frequency linear transducer for tumor invasion depth by a radiologist. In each case, both imaging findings of MRI and US were evaluated independently for tumor detection and invasion depth by consensus of two radiologists and were compared the diagnostic accuracy between two imaging modalities according to the histopathologic findings. MR imaging with five MR pulse sequences, spin echo T1 and in- and out-of phase gradient echo T1 weighted images, FSE and SSFSE T2 weighted images, were performed. Five MR pulse sequences were evaluated and compared on the point of detection and accurate distinction of tumor from surrounding normal tissue. Results : In EGC, diagnostic accuracy of US(77%) was superior than that of MRI(59%) but no statistically significant difference was noted between two imaging modalities(p=0.096). In AGC, both imaging modalities of MRI and US showed relatively high diagnostic accuracy as 97% and 84% respectively. Diagnostic accuracy of MRI was statistically better than that of US at the significant level(p<0.001). The best MR pulse sequence among five in each specimen was FSE T2WI(75.5%, 40/53) in both EGC and AGC. In AGC, FSE T2WI showed excellent imaging quality by showing very high ratio (93.5%, 29/31) of accurate delineation of tumor. Conclusion : MRI and US show relatively high diagnostic accuracy in the evaluation of tumor invasion depth of resected specimen in AGC. The most excellent pulse sequence of MRI for the evaluation of tumor invasion depth is FSE T2WI on the point of detection and accurate delineation of tumor in both EGC and AGC.

  • PDF

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

  • 양덕춘;김용환;임학태;방극수;배창휴
    • Korean Journal of Plant Tissue Culture
    • /
    • v.28 no.3
    • /
    • pp.165-171
    • /
    • 2001
  • New visible and fast assay system have been developed for tobacco transformant introduced with adenosine deaminase (ADA) marker gene, which converts cytotoxic adenosine analogues to non-toxic inosine analogues and ammonia. Ammonia was changed to blue color in the solution of phenol-nitoprusside and alkaline-hypochlorite. It was possible to detect activity of ADA visibly on the holes of 96 well plate using tiny explant of transgenic tobacco leaves within 1 hour incubation time. As substrates of ADA enzyme from transgenic plant on the plate, a number of adenosine analogues such as 9-D-arabinofuranosyl adenine, cordycepin, 2'-deoxyadenosine, adenosine and xylofuranosyl adenine were possible for detection of ADA activity. Optimal condition of substrate for ADA enzyme was each 10 mM and pH 7.5 in adenosine solution. Especially, transgenic plant did not convert adenosine to inosine and ammonia in the presence of ADA inhibitor deoxycoformycin, which means that ammonia produced from transgenic plant is due to expression of ADA gene. Now, we show that this detection system can be easily, sensitively, fast and cheaply as well as visibly assayed in vitro as GUS gene system with very small size of transformant explant.

  • PDF

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

  • 한상미;이광길;여주홍;권해용;우순옥;이용우;백하주;한명세
    • Journal of Sericultural and Entomological Science
    • /
    • v.45 no.2
    • /
    • pp.103-107
    • /
    • 2003
  • Entomopathogenic nematodes were isolated through the investigation in soils collected from cultivated and non-cultivated fields using silkworms (Bombyx mori) and Galleria mellonella trap. The detectable rate of entomopathogenic nematodes of silkworms trap was higher than the G. mellonella trap. This study indicates the detection of entomopathogenic nematodes from soils that silkworms are sensitive superior to the G. mellonella to entomopathogenic nematodes. The steinernema, rhabditidae, and diplogatroidae strains successfully cultured on the silkworms host as well as on artificial media. Reproductivity in the living silkworm larva and pupa was 1.5 to 3.5${\times}$ 10$\^$5/ nematodes per host However, G. mellonella could be multiplied less than 5${\times}$l0$\^$5/ nematodes. The dried pupa of the silkworm following mositurize was cultured 0.5 to 2${\times}$10$\^$5/) nematodes per host. The culture methods of the steinernema, rhabditidae, and diplogatroidae strains, using silkworm powder, extracted chicken intestine, and food waste fertilizer could be applicative, but rate of reproduction was low.

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

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • Environmental Mutagens and Carcinogens
    • /
    • v.21 no.1
    • /
    • pp.34-43
    • /
    • 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.

  • PDF

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
    • Journal of Embryo Transfer
    • /
    • v.32 no.3
    • /
    • pp.105-109
    • /
    • 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
    • /
    • v.15 no.1
    • /
    • pp.93-100
    • /
    • 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
    • Korean Journal of Veterinary Research
    • /
    • v.52 no.4
    • /
    • pp.259-262
    • /
    • 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
    • Proceedings of the SCSK Conference
    • /
    • 2003.09a
    • /
    • pp.85-96
    • /
    • 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.

  • PDF

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

  • 신해식
    • Proceedings of the Korea Society of Environmental Toocicology Conference
    • /
    • 2004.05a
    • /
    • pp.47-60
    • /
    • 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.

  • PDF

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

  • Kim Jin Gyu;Sin Hae Sik
    • Proceedings of the Korea Society of Environmental Biology Conference
    • /
    • 2004.05a
    • /
    • pp.1-15
    • /
    • 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.

  • PDF