• 제목/요약/키워드: fluorescent in situ hybridization (FISH)

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Circulating Aneuploid Cells Detected in the Blood of Patients with Infectious Lung Diseases

  • Kim, Hongsun;Cho, Jong Ho;Sonn, Chung-Hee;Kim, Jae-Won;Choi, Yul;Lee, Jinseon;Kim, Jhingook
    • Journal of Chest Surgery
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    • 제50권2호
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    • pp.126-129
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    • 2017
  • The identification of circulating tumor cells (CTCs) is clinically important for diagnosing cancer. We have previously developed a size-based filtration platform followed by epithelial cell adhesion molecule immunofluorescence staining for detecting CTCs. To characterize CTCs independently of cell surface protein expression, we incorporated a chromosomal fluorescence in situ hybridization (FISH) assay to detect abnormal copy numbers of chromosomes in cells collected from peripheral blood samples by the size-based filtration platform. Aneuploid cells were detected in the peripheral blood of patients with lung cancer. Unexpectedly, aneuploid cells were also detected in the control group, which consisted of peripheral blood samples from patients with benign lung diseases, such as empyema necessitatis and non-tuberculous mycobacterial lung disease. These findings suggest that chromosomal abnormalities are observed not only in tumor cells, but also in benign infectious diseases. Thus, our findings present new considerations and bring into light the possibility of false positives when using FISH for cancer diagnosis.

소양호 세균군집구조와 그 구조에 영향을 주는 환경요인 (The Bacterial Communities Structure and Its Environmental Determinants in Lake Soyang)

  • 김동주;홍선희;최승익;안태석
    • 미생물학회지
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    • 제36권2호
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    • pp.136-141
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    • 2000
  • 소양호에서 수온, 엽록소$\alpha$, pH 그리고 DOC의 농도가 세균군집에 미치는 영향을 파악하였다. 이 연구에서는 fluorescent in situ hybridization(FISH) 방법을 사용하기 1998년 4월부터 5월까지 세균군집구조를 조사하였고 이 시기 환경요인과 세균군집간의 상관성 분석을 실시하기 위해 수온, 엽록체$\alpha$, pH, DOC의 측정도 함께 수행하였다. 또한 환경요인과 세균군집간의 상관성 분석을 실시하기 위해 통계프로그램(SPSS)을 이용하였다. 그 결과 소양호에서 DOC, 엽록체$\alpha$, pH와 온도가 다중선형희귀직선상 설명가능정도가 약 43~58%정도로 나타났으며, 세균군집변화의 약 50%정도가 본 조사 이외의 환경요인에 영향을 받은 것으로 나타났다. $\alpha$-group은 DOC가 증가할수록 감소하여 DOC와 음의 상관관계를 보였고 , ${\gamma}$-group은 엽록소 $\alpha$농도와 음의 상관관계를 나타내었다. $\beta$-Cytophaga-Flavobacterium group 의 경우 수온과 뚜렷한 양의 상관성이 있는 것으로 나타났다. 또한 $\alpha$-group과 Cytophaga-Flavobacterium group은 pH와 양의 상관관계를 보였다. 이처럼 소양호의 세균군집은 계절에 따른 여러 가지 수 환경요인의 변화에 의해 다양하게 변화하였다.

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Screening for down syndrome using trophoblast retrieval and isolation of the cervix: preliminary study

  • Lee, Min Jin;Kim, Soo Hyun;Park, Hee Jin;Shim, Sung Han;Jang, Hee Yeon;Cha, Dong Hyun
    • Journal of Genetic Medicine
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    • 제17권2호
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    • pp.68-72
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    • 2020
  • Purpose: Trisomy 21, the cause of Down syndrome (DS) with various medical problems, is the most common aneuploidy during the fetal period. For diagnosis, a non-invasive screening test using maternal blood, which cannot be confirmed and invasive confirmation test with a risk of miscarriage, may be performed. The trophoblast retrieval and isolation of the cervix (TRIC) have been proposed by some researchers as an alternative to overcome the limitations of current tests. We experimented using TRIC to identify the possibility of trisomy 21 for the first time in Asia. Materials and Methods: Three cases of DS were analyzed confirmed by invasive tests (chorionic villus sampling, amniocentesis). All samples of trophoblasts immediately were immersed in phosphate-buffered saline and processed with formalin for fixation. The trophoblasts were isolated using an anti-human leukocyte antigen-G antibody coupled to magnetic nanoparticles. β-human chorionic gonadotropin (hCG)-expressing cells were considered as trophoblast cells, and the detection rate calculated. DS was confirmed by fluorescence in situ hybridization (FISH). Results: The mean trophoblast detection rate using β-hCG was 78.1%, and the detection rate using FISH was 22.2%. In all cases, the trisomy of chromosome 21 was identified. Conclusion: Trophoblast can be obtained from the five weeks of gestation and has a high detection rate, so it is noted that it can replace the current prenatal genetic test. To realize the clinical application as a prenatal genetic test, we will need additional efforts to identify trisomy 21 as well as other chromosomal abnormalities in future large-scale studies.

형광직접보합법을 이용한 미배양 양수세포에서 산전 이수배수체 확인 (Prenatal Aneuploidy Detection in Uncultured Amniotic Fluid Interphase Cells by Fluorescence in situ Hybridization (FISH))

  • 설혜원;고희정;송남희;김숙령;이화진;오선경;박중신;전종관;윤보현;신희철;문신용
    • Clinical and Experimental Reproductive Medicine
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    • 제30권3호
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    • pp.223-231
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    • 2003
  • Objective: The aim of the present study was to evaluate the clinical efficiency of fluorescent in situ hybridization (FISH) in the prenatal diagnosis of chromosomal aneuploidy. Methods: We reviewed data of 268 cases to identify women undergoing genetic amniocentesis at cytogenetic laboratory, from January 2000 to December 2002. Amniotic fluid was submitted for both rapid FISH on uncultured interphase amniocytes using a commercially available DNA probe for chromosome 13, 18, 21, X, Y and standard karyotyping on cultured metaphase amniocytes. Results from FISH and full karyotype were compared. Results: There were 251 cases (84%) normal and 17 cases (16%) abnormal in FISH results. All 17 cases of trisomy 13, 18, 21 including two cases of mosaicism and sex chromosome aneuploidies which are detected by FISH were confirmed with conventional cytogenetics and there was no false positive result. Twenty two cases had karyotypically proven abnormalities that could not have been detected by the targeted FISH. Conclusion: Interphase FISH analysis of uncultured amniotic fluid cells has been shown to be an effective and reliable technique for rapid fetal aneuploidy screening during pregnancy as an adjunctive test to conventional cytogenetics.

활성슬러지내의 전기화학적활성 박테리아 분포 특성 (Distribution of Electrochemically Active Bacteria in Activated Sludge Characteristics)

  • 손형식;손희종;김미아;이상준
    • KSBB Journal
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    • 제26권5호
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    • pp.407-411
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    • 2011
  • Microbial fuel cell (MFC) wes enriched using sludge in wastewater treatment. The microbial community of activated sludge and enriched MFC were analyzed by FISH (fluorescent in situ hybridization) and 16S rDNA sequencing. Bacteroidetes group were pre-dominant in activated sludge by FISH. ${\alpha}$ group, ${\gamma}$ group and Acintobacter group were dominant and they were similar to distribution. The average value of 10 peak of MFC is 0.44C. When MFC wase enriched by sludge, ${\gamma}$-Proteobacteria, Plantomycetes group increased 70% and 60%, respectively. In results of 16S rDNA sequencing, Sphiringomonas sp. was comprised in ${\alpha}$ proteobacteria and Enterobacter sp., Klebsiella sp., Acinetobacter sp., Bacillus sp. were comprised in ${\gamma}$ proteobacteria and Chryseobacterium sp. was comprised in Flavobacteria were isolated from sludge.

남매지에서 Eubacteria 군집구조의 계절적 변화와 그에 영향을 미치는 환경요인 (Seasonal Variation of Eubacterial Community Structure and Their Structure Affecting Environmental Parameters in Reservoir)

  • 이희순;박정원;김미경;이영옥
    • 미생물학회지
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    • 제38권1호
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    • pp.31-37
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    • 2002
  • 경북 경산시 에 위치한 남매지 에서 세균군집 의 구조를 fluorescent in situ hybridization(FISH) 방법을 이용하여 분석하였고 아울러 세균군집크기에 영향을 주는 환경요인을 측정하여 그 상관성을 파악하고자 하였다. 담수계에 우점하는 것으로 알려진 proteobacteria($\alpha$.$\beta$.$\gamma$-subclasses)와 글Cytophaga-Flavobacterium (CF) group에 속하는 진정세균군의 연중변화를 정점별로 측정하였다. 총세균수에 대한 각 세균군집 비율은 정점간의 큰 차이 없이 $\alpha$-subclass가 4.0~29.2%,$\beta$-subclass가 1.7~25.8%, $\gamma$-subclass가 1.8~12.8%를, 그리고 CF group은 4.9~36.3%의 군집 비율을 나타냈다. 온도,SS, pH, DOC,질산염, 조류 현존량 등 환경요인과 세균군집 간의 상관관계를 분석한 결과, 총세균수는 수온, DOC,55, eubacteria는 DOC, 녹조류 현존량, $\gamma$-subclass 세균군은 DOC, CF group세 균은 녹조류 현존량과 양의 상관성을 나타낸 반면 $\beta$-subclass는 남세균 . 총 조류 현존량과 음의 상관성을 나타냈다.

소양호에서 세균군집구조의 계절적.수직적 변화 (Seasonal and Vertical Change of Bacterial Communities in Lake Soyang)

  • 김동주;홍선희;안태석
    • 미생물학회지
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    • 제35권3호
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    • pp.242-247
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    • 1999
  • 소양호에서 fluorescent in situ hybridization 방법을 이용하여 세균군집의 계절과 수심에 따른 변화를 조사하였다. 이 방법에 사용된 oligonucleotide probe는 EUB338, ALF1b, BET42a, GAM42a 와 CF probe 였다. 조사기간중 Proteobacteria $\alpha$-group 은 0.7~32.9% 이었으며 $\beta$-group은 1.0~25.8%, $\gamma$-group은 2.4~37.1%, Cytophaga-Flavobacterium group은 4.7~23.6%의 분포를 나타내었다. 계절별로 살펴보면 규조류가 우점하는 봄펄에는 $\gamma$-group 이, 유기물의 농도가 낮고 수온이 높은 여름철에는 $\alpha$-group 이 우점하였으며, 남조류가 우점하는 가을철에 총세균수에 대한 Eubacterial group 의 비율이 크게 감소하여 특정 group의 우점현상은 나타나지 않았다. 이처럼 소양호는 계절과 수심에 따라 군집구조가 변화하였으며, 특히 세균의 군집구조는 식물플랑크톤의 천이와 밀접한 관계를 보이는 것으로 나타났다.

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음식물 쓰레기와 폐활성 슬러지를 이용한 생물학적 수소생산 및 수소생산 미생물 군집분석 (Continuous Bio-hydrogen Production from Food Waste and Waste Activated Sludge)

  • 김동건;이윤지;김동임;김지성;유명진;박대원;김미선;상병인
    • KSBB Journal
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    • 제20권6호
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    • pp.438-442
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    • 2005
  • 1. 회분식 실험결과 유기물의 함량이 높은 음식물만을 기질로 이용한 경우보다 폐활성 슬러지의 혼합비율이 $10{\sim}20%$일 때 더 높은 수소생산을 나타내었다. 또한 폐활성 슬러지의 혼합비율이 40%인 경우에는 메탄이 발생하여 생성된 수소가 소모되는 반응을 나타내었다. 2. 연속 실험의 경우 HRT를 줄여 유기물의 부하를 증가시킬 경우 수소생산량이 급격히 증가하였으며, HRT 2일까지는 미생물의 wash out 없이 안정적 수소생산을 보였다. 3. 음식물과 폐활성 슬러지를 이용한 연속 운전을 HRT와 두 기질의 비율을 달리하여 운전한 결과 2일의 HRT와 FW:WAS=80:20의 비율에서 140 mL $H_2/g$ VSS의 높은 수소생산율을 얻을 수 있었다. 4. 음식물 쓰레기와 폐활성 슬러지의 비율을 적절히 혼합할 경우, 수소생산의 상승작용을 할 수 있는 가능성을 확인하였다. 5. SEM과 FISH 분석을 통하여 반응조 내의 수소 미생물의 공간적 분포 및 형태를 관측하였으며, 음식물이나 슬러지 주변에 많은 수소생산 미생물이 관측되었다.

Visualization of chromatin higher-order structures and dynamics in live cells

  • Park, Tae Lim;Lee, YigJi;Cho, Won-Ki
    • BMB Reports
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    • 제54권10호
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    • pp.489-496
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    • 2021
  • Chromatin has highly organized structures in the nucleus, and these higher-order structures are proposed to regulate gene activities and cellular processes. Sequencing-based techniques, such as Hi-C, and fluorescent in situ hybridization (FISH) have revealed a spatial segregation of active and inactive compartments of chromatin, as well as the non-random positioning of chromosomes in the nucleus, respectively. However, regardless of their efficiency in capturing target genomic sites, these techniques are limited to fixed cells. Since chromatin has dynamic structures, live cell imaging techniques are highlighted for their ability to detect conformational changes in chromatin at a specific time point, or to track various arrangements of chromatin through long-term imaging. Given that the imaging approaches to study live cells are dramatically advanced, we recapitulate methods that are widely used to visualize the dynamics of higher-order chromatin structures.