• 제목/요약/키워드: Fluorescence In Situ Hybridization

검색결과 206건 처리시간 0.03초

혐기 혼합균주에서 황산염 농도변화에 따른 수소 발효 특성 (Fermentative Hydrogen Production under Various $SO_4^{2-}$ Concentration using Anaerobic Mixed Microflora)

  • 황재훈;최정아;이종학;정태영;차기철;송호철;용보영;김동진;전병훈
    • 대한환경공학회지
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    • 제31권6호
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    • pp.434-441
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    • 2009
  • 황산염의 농도변화에 따른 연속 혐기성 수소 발효에 미치는 영향을 고찰하기 위해서 혼합균주를 사용한 완전 혼합형 반응조를 운전하였다. 기질은 글루코오스를 사용하였고, 수리학적 체류시간은 1, 0.5, 0.25 일로 각각 고정하였다. 황산염 농도는 0${\sim}$20,000 mg/L로 단계별 증가시켰고 pH 5.5로 운전하였다. 실험 결과 높은 황산염 농도에 관계없이 수소가 발생하였고, HRT 0.25일로 짧아짐에 따라 수소 발생이 높게 나타났다. HRT 1, 0.5, 0.25일 각 조건별 수소 생성량과 수소 수율은 2.9, 4.6, 9.4 L/day, 2.0, 1.8, 1.6 mol $H_2$/mol glucose로 나타났으며, 잔존 황산염 96${\sim}$98, 95${\sim}$97 94${\sim}$97%로 나타나 황산염 환원이 발생하지 않았다. FISH 결과 모든 조건에서 수소생성균의 분포는 나타났지만 황산염환원균의 분포는 나타나지 않았다.

Efficacy of Primed In Situ Labelling in Determination of HER-2 Gene Amplification and CEN-17 Status in Breast Cancer Tissue

  • Salimi, Mahdieh;Mozdarani, Hossein;Majidzadeh-A, Keivan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권1호
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    • pp.329-337
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    • 2012
  • Considerable attention has been given to the accuracy of HER-2 testing and the correlation between the results of different testing methods. This interest reflects the growing importance of HER-2 status in the management of patients with breast cancer. In this study the detection of HER-2 gene and centromere 17 status was evaluated using dual-colour primed in situ labelling (PRINS) in comparison with fluorescence in situ hybridization (FISH). These two methods were evaluated on a series of 27 formalin fixed paraffin embedded breast carcinoma tumours, previously tested for protein overexpression by HercepTest (grouped into Hercep 1+/0, 2+ and 3+). HER-2 gene amplification (ratio${\geq}2.2$) by PRINS was found in 3:3, 6:21 and 0:3 in IHC 3+, 2+ and 1+/0 cases, respectively. Comparing FISH and IHC (immunohistochemistry), showed the same results as for PRINS and IHC. Chromosome 17 aneusomy was found in 10 of 21 IHC 2+ cases (47.6%), of which 1 (10%) showed hypodisomy (chromosome 17 copy number per cell${\leq}1.75$), 7 (70%) showed low polysomy (chromosome 17 copy number per cell=2.26 - 3.75) and 2 (20%) showed high polysomy (chromosome 17 copy number per cell ${\geq}3.76$). The overall concordance of detection of HER-2 gene amplification by FISH and PRINS was 100% (27:27). Furthermore, both the level of HER-2 amplification and copy number of CEN17 analysis results correlated well between the two methods. In conclusion, PRINS is a reliable, reproducible technique and in our opinion can be used as an additional test to determine HER-2 status in breast tumours.

Invertase 분석이 통합된 sandwich hybridization에 의한 Cochlodinium polykrikoides의 신속 검출 (Rapid Detection of Cochlodinium polykrikoides by Sandwich Hybridization Integrated with Invertase Assay)

  • 강민경;박미례;이택견
    • 한국산학기술학회논문지
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    • 제20권8호
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    • pp.627-633
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    • 2019
  • 유해조류 대발생(harmful algal blooms, HABs)을 유발하는 해양 미세조류의 신속하고 정확한 종판별은 HABs을 예측하고 관리하기 위한 매우 중요한 도구이다. 우리는 이전 연구에서 적조 유발 미세조류인 Cochlodinium polykrikoides를 현장에서 검출하기 위한 nuclease protection assay sandwich hybridization (NPA-SH) 방법을 개발한 바 있다. 본 연구에서는 C. polykrikoides 검출을 위한 NPA-SH 방법의 반응 단계를 간소화시켜 현장 적용성을 향상시키는 것을 목적으로, signal probe에 형광 대신 invertase (INV)를 결합시켰으며, hybridization 과정에서 sucrose를 반응물로 사용하여 발색 반응을 유도하였다. INV와 signal probe의 결합 여부는 SDS-PAGE와 형광 현미경을 사용하여 확인하였으며, 반응 최적화를 위한 적정 프로브의 양, sucrose의 양 및 처리시간 등을 정립하였다. 본 연구의 결과 개발된 INV-SH는 NPA-SH와 비교하여 현장에서의 처리시간이 감소되었으며, 흡광도 측정기 뿐만 아니라, 상대적으로 부피가 작고 값 싼 개인 혈당계 사용이 가능하게 되었다. 본 연구에서 개발된 INV-SH는 INV가 적용된 최초의 C. polykrikoides 종판별 기술이며, 증진된 현장기술이 될 수 있다.

성장부진과 발달지연을 보인 환아에서 확인된 환상 9번 염색체 1례의 세포유전학적인 연구 (Cytogenetic evaluation of a patient with ring chromosome 9 presenting failure to thrive and developmental delay)

  • 박윤미;노한내;김숙자;안영민
    • Clinical and Experimental Pediatrics
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    • 제51권4호
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    • pp.426-430
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    • 2008
  • 환상 염색체의 발생 기전은 염색체 말단 부분이 결손된 후 양 끝이 융합되거나 끝분절 염기서열이 앞뒤역순상동서열로 융합될 경우로 생각되고 있다. 환상 염색체는 세포 분열을 하는 동안 불안정하기 때문에 세포핵이 없는 딸세포의 사망률이 증가하게 되어 생존하는 세포수가 감소하고 성장 장애가 발생하게 된다. 표현형은 염색체 손실의 정도에 따라 다양하다. 저자들은 최초로 심각한 저신장을 주소로 내원한 환아의 염색체 검사상 환상 9번 염색체를 확인하였기에 이를 보고하는 바이다.

5S와 45S rDNA 유전자를 이용한 제주도산 애기더덕 (Codonopsis minima)과 더덕 (C. lanceolata)의 FISH 패턴 분석 (Analysis of FISH patterns using 5S and 45S rDNAs in Codonopsis minima and C. lanceolata from Jeju Island)

  • 김수영;김찬수
    • 한국약용작물학회지
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    • 제18권3호
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    • pp.186-190
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    • 2010
  • The chromosome number was identified and fluorescence in situ hybridization(FISH) mapping of 5S and 45S rDNAs were conducted for C. minima and C. lanceolata in the genus Codonopsis from Jeju island. In this study, we have confirmed that the somatic metaphase chromosome number determined as 2n=2x=16 was the same as the findings from the previous studies. While the conventional staining method makes it rather difficult to distinguish satellite chromosomes due to high degree of variability, FISH analysis produced the exact number and location of 5S and 45S rDNAs. Both species in the genus Codonopsis have a pair of 5S rDNA and their gene loci were observed on chromosome 3. Although two pairs of 45S rDNAs (one on chromosome 1 and the other on chromosome 8) were identified in both species, the 45S rDNA signals on chromosome 8 in C. minima were significantly weaker than those on chromosome 1. In addition, the 45S rDNA signals on chromosome 1 in C. lanceolata showed that the chromosome is non-homologus. In this study, we have determined cytogenetic characteristics of C. minima and C. lanceolata according to their gene replication patterns.

마이크로어레이 분석기법의 임상적용에 관한 연구 (Medical Implementation of Microarray Technology)

  • 강지언
    • 대한임상검사과학회지
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    • 제52권4호
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    • pp.310-316
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    • 2020
  • 마이크로어레이 진단 기법의 발달은 세포유전학적 관점에서, 다양한 종류의 유전학적 질병과 관련하여 새로운 정보를 제공하고, 질병에 대한 기본적인 통찰력을 제공하는데 매우 중요한 역할을 제공하고 있다. 그동안 많은 연구들에서, 마이크로어레이 기술을 활용한 인간 게놈의 유동성과 다양성을 입증해 주었으며, 게놈의 취약성을 식별하기 위한 보다 정확한 진단기법과 적절한 임상 관리 방법을 효율적으로 제공해 왔다. 앞으로 다양한 유전과 관련된 질병에 기존 세포유전학적 방법을 자동화된 마이크로어레이 방법으로 전환한다면, 보다 효율적인 방법으로 질병을 진단하고, 정확성을 향상시키며, 유전자 배열의 암호화 및 복잡한 특성을 밝히는데 매우 중요한 역할을 할 것으로 생각된다. 또한 이 분석 기법을 활용하여 게놈과 인간의 건강, 질병과의 관계를 분석하여 다양한 정보를 미리 제공하여 질병을 예방하고, 질병의 진단 및 치료에도 도움이 될 수 있는 새로운 혁명을 일으킬 수 있을 것으로 기대된다.

Molecular and Cytogenetic Analysis of Transgenic Plants of Rice(Oryza sativa L.) Produced by Agrobacterium-mediated Transformation

  • Cho, Joon-Hyeong;Kim, Yong-Wook
    • Plant Resources
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    • 제7권1호
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    • pp.39-46
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    • 2004
  • To demonstrate the importance of transformation efficiency in independent event, molecular and cytogenetic analysis were conducted with genomic DNA and chromosome of transgenic plants produced by Agrobacterium tumefeciens LBA4404 (pSBM-PPGN: gusA and bar). Selection ratios of putative transgenic calli were similar in independent experiments, however, transformation efficiencies were critically influenced by the type of regeneration media. MSRK5SS-Pr regeneration mediun, which contains 5 mgL$^{-1}$ kinetin, 2% (w/v) sucrose in combination with 3% (w/v) sorbitol, and 500 mgL$^{-1}$ proline, was efficient to produce transgenic plant of rice from putative transgenic callus in the presence of L-phosphinotricin (PPT). With MSRK5SS-Pr medium, transformation efficincies of Nagdongbyeo were significantly enhanced from 3.7% to 6.3% in independent callus lines arid from 7.3% to 19.7% in plants produced, respectively. Stable integration and expression of bar gene were confirmed by basta herbicide assay, PCR amplification and Southern blotting of bar gene, and fluorescence in situ hybridization (FISH) analysis using pSBM-PPGN as a probe. In Southern blot analysis, diverse band patterns were observed in total 44 transgenic plants regenerated from 20 independent PPT resistant calli showing from one to five copies of T-DNA segments, however, the transformants obtained from one callus line showed the same copy numbers with the same fractionized band patterns.

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SHARON/ANAMMOX 결합공정에서 슬러지의 입상화와 특성 (Granulation and Characteristics of Sludges in the Combined SHARON/ANAMMOX Processes)

  • 황인수;민경석
    • 한국물환경학회지
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    • 제22권2호
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    • pp.300-307
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    • 2006
  • The combined SHARON (Single reactor system for High ammonium Removal Over Nitrite)-ANAMMOX (Anaerobic ammonium oxidation) reactor were operated in mesophilic condition ($35^{\circ}C$). In this study, microbial granulation and characteristics of SHARON and ANAMMOX sludges were investigated using settling test, Scanning Electron Microscopy (SEM) and Fluorescence In Situ Hybridization (FISH). In SHARON reactor, Aerobic granulation with diameter of 1.5~2.5 mm was accomplished but aerobic granulation was weaker than anaerobic granular sludge. Initial seed sludge of ANAMMOX reactor was used as attached media for biofilm growth. ANAMMOX sludge was more compact and rounder rather than seed sludge. Though ANAMMOX sludge has high activity, it has lower settling ability than the seed granule. The color of ANAMMOX sludge was changed from dark to redish brown granular with diameter of 1~2 mm. In FISH of ANAMMOX sludge, high fraction of Candidatus B. stuttgartiensis which paid great role of nitrogen conversion was detected. Also, FISH results reveals that ANAMMOX bacteria inhabit at inner parts near surface, having advantages in utilization of substrates and protection from oxygen inhibition.

요 세포검사의 최근 검사기법 (New Techniques for the Detection of the Malignant Cells in Urine Cytology)

  • 공경엽
    • 대한세포병리학회지
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    • 제17권1호
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    • pp.18-26
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    • 2006
  • Transitional cell carcinoma of the urinary bladder is common in the genitourinary tract. The gold standard for the diagnosis of bladder cancer has been cystoscopy, along with urine cytology. Cystoscopy is an invasive and relatively expensive technique. By comparison, urine cytology is easy to perform and specific for a diagnosis of bladder cancer, although less sensitive, especially in low-grade tumors. For this reason, there has been a need for superior noninvasive technology to increase our confidence in being able to detect bladder cancer. There are many reports of the various urinary tests that are available to facilitate the diagnosis. In this article, I reviewed the literature on urinary markers and tests that may be clinically useful, including fluorescence in situ hybridization, uCyt+/Immunocyte, the $BTA^{(R)}$ test, the NMP 22TM, the $FDP^{(R)}$ test, the telomerase activity test, the HA and HAse tests, and flow cytometry. Most of these tests have a higher sensitivity and specificity than cytology. However, urine cytology has the highest specificity, especially in individuals with a high-grade tumor. We conclude that no urinary markers or tests can replace the role of cystoscopy along with cytology in the diagnosis of transitional cell carcinoma of the bladder. However, some markers could be used adjunctively to increase the diagnostic accuracy during screening or during the postoperative follow-up examination of patients with bladder cancer.

Upregulation by KCI Treatment of Eukaryotic Translation Elongation Factor 1A (eEF1A) mRNA in the Dendrites of Cultured Rat Hippocampal Neurons

  • Moon, Il Soo;Cho, Sun-Jung;Lee, HyunSook;Seog, Dae-Hyun;Jung, Yong Wook;Jin, IngNyol;Walikonis, Randall
    • Molecules and Cells
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    • 제25권4호
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    • pp.538-544
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    • 2008
  • Activity-dependent local translation in the dendrites of brain neurons plays an important role in the synapse-specific provision of proteins necessary for strengthening synaptic connections. In this study we carried out combined fluorescence in situ hybridization (FISH) and immunocytochemistry (IC) and showed that more than half of the eukaryotic elongation factor 1A (eEF1A) mRNA clusters overlapped with or were immediately adjacent to clusters of PSD-95, a postsynaptic marker, in the dendrites of cultured rat hippocampal neurons. Treatment of the neurons with KCl increased the density of the dendritic eEF1A mRNA clusters more than two-fold. FISH combined with IC revealed that the KCl treatment increased the density of eEF1A mRNA clusters that overlapped with or were immediately adjacent to PSD-95 clusters. These results indicate that KCl treatment increases both the density of eEF1A mRNA clusters and their synaptic association in dendrites of cultured neurons.