• Title/Summary/Keyword: fluorescence in-situ

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Influence of FA and FNA to Microbial Community in Sequencing Batch Ammonium Partial Nitrification System (암모니아 부분산화 공정의 제어와 미생물 군집의 변화)

  • Ahn, Johwan
    • Journal of Korean Society on Water Environment
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    • v.31 no.4
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    • pp.399-406
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    • 2015
  • A sequencing batch reactor was operated under different pH conditions to see the influence of free ammonia (FA) and free nitrous acid (FNA) to microbial community on ammonium partial nitrification. Long-term influences of FA and FNA were evaluated by polymerase chain reaction-denaturing gradient gel electrophoresis and fluorescence in situ hybridization. Nitrite accumulation was successfully achieved at pH 8.2 and 6.3. The shifts in the microbial community were observed when influent ammonia concentration increased to 1 g $NH_4$-N/L at pH 8.2, and then when pH was dropped to 6.3. Both Nitrosomonas and Nitrosospira were selected during the startup of the reactor, and eventually became dominant members as ammonia-oxidizing bacteria. The results of molecular microbiological analysis strongly suggested that the composition of microbial community was changed according to the method used to control nitrite-oxidizing bacteria.

혐기성 SBR을 이용한 anammox 미생물 배양 및 fluorescence in situ hybridization (FISH)을 통 미생물 군집 분석

  • Han, Dong-U;Yun, Ho-Jun;Kim, Dong-Jin
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.286-289
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    • 2001
  • Anaerobic ammonium oxidation with nitrite to $N_2$(anammox) is a recently discovered microbial reaction with interesting potential for nitrogen removal from wastewater. Here we investigated the microbial community structure in the sequencing batch reactor(SBR) with an anammox activity. The SBR was optimized for the enrichment of a very slowly growing microbial community and showed that possibility of anaerobic ammonium oxidation. Fluorescence in situ hybridization(FISH) analysis revealed that anaerobic ammonium oxidizers were Candidatus Brocadia anammoxidans and Candidatus Kuenenia stuttgartiensis. Furthermore, Nitrosomol1as spp. of the ${\beta}$ -subclass of Proteobacteria was also present within the anaerobic SBR microorganisms.

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A Case Study on Geotechnical Properties and Weathering Degree of Weathered Granite Rock (화강 풍화암의 지반특성 및 풍화도 평가에 대한 사례 연구)

  • Lee, Seung-Hwan;Yoo, Byeong Soo;Chung, Choong-Ki
    • Journal of the Korean Geotechnical Society
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    • v.33 no.12
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    • pp.127-139
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    • 2017
  • Site investigation including boring and various in-situ borehole test (Pressuremeter test, Borehole shear test, Downhole test, Suspension PS logging, Density logging) and X-ray fluorescence analysis for rock core sample were performed to estimate geotechnical properties and weathering degree of weathered granite rock in Goyang. Deformation modulus, shear strength parameter and shear wave velocity estimated through in-situ borehole test had a tendency to increase with depth. And several chemical weathering indices evaluated by X-ray fluorescence analysis had a general tendency of reducing weathering degree in accordance with depth. Also, relationship between VR determined as a representative weathering index and the geotechnical properties was analyzed.

In-Situ Observation of New Extra-Vascular Threadlike Structure of Mouse Using a Fluorescence Stereoscopic Microscope

  • Sung, Baeck-Kyoung;Lee, Ja-Woong;Lee, Byung-Cheon;Johng, Hyun-Min;Baik, Ku-Youn;Nam, Tae-Jeong;Park, Dae-Hun;Soh, Kyeong-Sun;Soh, Kwang-Sup
    • Journal of Pharmacopuncture
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    • v.7 no.3
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    • pp.73-76
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    • 2004
  • We report the new threadlike structures outside the blood vessels of mice. For this, we developed an in-situ searching method of the structure by vital staining with the dye of acridine orange and using a fluorescent stereomicroscope designed specifically for this purpose. We consider that the newly found threadlike structure might be rediscovery of the extra-vascular Bonghan duct which was reported in 1963 by Bonghan Kim.

Design, Optimization and Verification of 16S rRNA Oligonucleotide Probes of Fluorescence in-situ Hybridization for Targeting Clostridium spp. and Clostridium kluyveri

  • Hu, Lintao;Huang, Jun;Li, Hui;Jin, Yao;Wu, Chongde;Zhou, Rongqing
    • Journal of Microbiology and Biotechnology
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    • v.28 no.11
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    • pp.1823-1833
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    • 2018
  • Fluorescence in-situ hybridization (FISH) is a common and popular method used to investigate microbial communities in natural and engineered environments. In this study, two specific 16S rRNA-targeted oligonucleotide probes, CLZ and KCLZ, were designed and verified to quantify the genus Clostridium and the species Clostridium kluyveri. The optimal concentration of hybridization buffer solution for both probes was 30% (w/v). The specificity of the designed probes was high due to the use of pellets from pure reference strains. Feasibility was tested using samples of Chinese liquor from the famed Luzhou manufacturing cellar. The effectiveness of detecting target cells appears to vary widely in different environments. In pit mud, the detection effectiveness of the target cell by probes CLZ and KCLZ was 49.11% and 32.14%, respectively. Quantitative analysis by FISH technique of microbes in pit mud and fermented grains showed consistency with the results detected by qPCR and PCR-DGGE techniques, which showed that the probes CLZ and KCLZ were suitable to analyze the biomass of Clostridium spp. and C. kluyveri during liquor fermentation. Therefore, this study provides a method for quantitative analysis of Clostridium spp. and C. kluyveri and monitoring their community dynamics in microecosystems.

The Optimization of Human Sperm Decondensation Procedure for Fluorescence in Situ Hybridization (Fluorescence in Situ Hybridization 시행을 위한 인간정자 탈응축법의 적정화)

  • Pang, Myung-Geol
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.3
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    • pp.369-375
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    • 1997
  • Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West et al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS). Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at $37^{\circ}C$ for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at $4^{\circ}C$. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious "nullisomy" due to hybridization failure without inducing spurious "disomy" resulting from increased distances between split signals.

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Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH) (Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색)

  • 방명걸
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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Fluorescence-in-situ-hybridization in the Surveillance of Urothelial Cancers: Can Use of Cystoscopy or Ureteroscopy be Deferred?

  • Ho, Christopher Chee Kong;Tan, Wei Phin;Pathmanathan, Rajadurai;Tan, Wei Keith;Tan, Hui Meng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4057-4059
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    • 2013
  • Background: Fluorescence in situ hybridization (FISH) testing may be useful to screen for bladder carcinoma or dysplasia by detecting aneuploidy chromosomes 3, 7, 17 and deletion of the chromosome 9p21 locus in urine specimens. This study aimed to assess the sensitivity, specificity, positive and negative predictive value of FISH in a multi-ethnic population in Asia. Materials and Methods: Patients with haematuria and/or past history of urothelial cancer on follow-up had their voided urine tested with FISH. Patients then underwent cystoscopy/ureteroscopy and any lesions seen were biopsied. The histopathological reports of the bladder or ureteroscopic mucosal biopsies were then compared with the FISH test results. Results: Two hundred sixty patients were recruited. The sensitivity and specificity of the FISH test was 89.2% and 83.4% respectively. The positive (PPV) and negative predictive values (NPV) were 47.1% and 97.9%. By excluding patients who had positive deletion of chromosome 9, the overall results of the screening test improved: sensitivity 84.6%; specificity 96.4%; PPV 75.9% and NPV 97.9%. Conclusions: UroVysion FISH has a high specificity of detecting urothelial cancer or dysplasia when deletion of chromosome 9 is excluded. Negative UroVysion FISH-tests may allow us to conserve health resources and minimize trauma by deferring cystoscopic or ureteroscopic examination.

1p36 deletion syndrome confirmed by fluorescence in situ hybridization and array-comparative genomic hybridization analysis

  • Kang, Dong Soo;Shin, Eunsim;Yu, Jeesuk
    • Clinical and Experimental Pediatrics
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    • v.59 no.sup1
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    • pp.14-18
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    • 2016
  • Pediatric epilepsy can be caused by various conditions, including specific syndromes. 1p36 deletion syndrome is reported in 1 in 5,000-10,000 newborns, and its characteristic clinical features include developmental delay, mental retardation, hypotonia, congenital heart defects, seizure, and facial dysmorphism. However, detection of the terminal deletion in chromosome 1p by conventional G-banded karyotyping is difficult. Here we present a case of epilepsy with profound developmental delay and characteristic phenotypes. A 7-year-and 6-month-old boy experienced afebrile generalized seizure at the age of 5 years and 3 months. He had recurrent febrile seizures since 12 months of age and showed severe global developmental delay, remarkable hypotonia, short stature, and dysmorphic features such as microcephaly; small, low-set ears; dark, straight eyebrows; deep-set eyes; flat nasal bridge; midface hypoplasia; and a small, pointed chin. Previous diagnostic work-up, including conventional chromosomal analysis, revealed no definite causes. However, array-comparative genomic hybridization analysis revealed 1p36 deletion syndrome with a 9.15-Mb copy loss of the 1p36.33-1p36.22 region, and fluorescence in situ hybridization analysis (FISH) confirmed this diagnosis. This case highlights the need to consider detailed chromosomal study for patients with delayed development and epilepsy. Furthermore, 1p36 deletion syndrome should be considered for patients presenting seizure and moderate-to-severe developmental delay, particularly if the patient exhibits dysmorphic features, short stature, and hypotonia.