• Title/Summary/Keyword: Fluorescence In situ Hybridization

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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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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.

Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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Fluorescence in situ hybridization using 16S rRNA-targeted oligonucleotides reveals abundance and spatial organization of methanogens in thermophilic sludge granules

  • Lee, Yu-Jin;Kim, Hyo-Seop;An, Yeong-Hui;Park, Seong-Hun
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.508-511
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    • 2000
  • In situ hybridization with fluorescent oligonucleotides(FISH) was used to detect and localize microorganisms in the granules of lab-scale upflow anaerobic sludge blanket(UASB) reactors. An UASB reactor was seeded with mesophilically-grown($35^{\circ}\;C$) granular sludge, and thermophilically($55^{\circ}\;C$) operated by feeding with a synthetic wastewater. Sections of the granules were hybridized with 16S rRNA-targeted oligonucleotide probes for Eubacteria, Archaeabacteria, and specific phylogenetic groups of methanogens. FISH clearly showed the layed structure of thermophilic granules, which was consisted of outer bacterial cells and inner archaeal cells. Methanoseata-, Methanosarcina-like cells were also found to be localized inside the granules. These results demonstrated FISH was useful in studying the spatial organizations of methanogens and in situ morphologies and metabolic functions in thermophilic granular sludges.

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Fluorescence In Situ Hybridization Analysis for Polysomy of Chromosome 17 in Head and Neck Adenoid Cystic Carcinomas (두경부 선양낭성암종에서 형광동소결합을 이용한 제17번 염색체의 다염색체 소견)

  • Choi Geon;Park Jae-Hyung;Choi Choong-Sik;Song Jae-Joon;Jung Kwang-Yoon;Choi Jong-Ouck
    • Korean Journal of Head & Neck Oncology
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    • v.16 no.1
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    • pp.3-8
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    • 2000
  • Background and Objectives: Adenoid cystic carcinoma of salivary glands is characterized by insidious growth over many years, local recurrences, and distant metastasis and classified to three distinct histologic subtypes: tubular, cribriform, and solid. The solid type is known to have the worst prognosis. However, histopathologic heterogeneity is observed in tumors from the same patient. We have attempted to elucidate the genotypic differences, characterized by polysomies of chromosome 17, in adenoid cystic carcinoma according to the phenotypic histopathologic heterogeneity. Materials and Methods: Fluorescence in situ hybridization was performed on formalin-fixed paraffin blocks from seven patients with head and neck adenoid cystic carcinoma, using the centromeric $\alpha$-satellite probe of chromosome 17 to detect nuclei exhibiting polysomy. The difference in polysomeric chromosome expression in cribriform, tubular, solid type and type I, II, III according to the Szanto classification was analyzed. Results: Polysomy of chromosome 17 was found in 15.28% of the cribriform type, in 15.68% of the tubular type, and in 18.87% of the solid type. The proportion of polysomy was statistically higher in the solid type than in the cribriform type(p<0.05), and the proportion of polysomy increased progressively from type 1 to type 3, but this trend was statistically insignificant(p>0.05). Conclusion: We suggest that there may be genetic variations in tumor from the same patient depending on the histopathologic heterogenetiy in adenoid cystic carcinomas.

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Determination of HER2 Gene Amplification in Breast Cancer using Dual-color Silver Enhanced in situ Hybridization (dc-SISH) and Comparison with Fluorescence ISH (FISH)

  • Unal, Betul;Karaveli, Fatma Seyda;Pestereli, Hadice Elif;Erdogan, Gulgun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6131-6134
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    • 2013
  • Background: The two basic methods that are currently accepted to identify the HER2 status are immunohistochemistry and flyorescence in situ hybridization (FISH). The aim of this study was to perform the dual-color silver in situ hybridization (dc-SISH) technique as an alternative to FISH. Materials and Methods: A total of 40 invasive breast carcinoma cases were assessed for HER2 gene amplification by FISH and dual-color SISH. Results: Significant correlation was found in the HER2 expression results obtained with the two approaches (p=0.001, p<0.05). The concordance rate was 92.3%. Conclusions: Foutine practical use of the dc-SISH method, which is much easier to apply, score, and evaluate, has many advantages. HER2 and CEN17 status can be evaluated simultaneously with the newly developed "Dual-Color Probe". All these specifications and the reliable results obtained support the widespread use of SISH technique in clinical practice.

Wastewater Treatment and Microbial Structure Analysis by Fluorescence In Situ Hydridizationin a Biofilm Reactor (생물막 반응키에서의 폐수 처리 및 Fluorescence In Situ Hybridization에 의한 복합 미생물계 구조 해석)

  • Kim, Dong-Jin;Han, Dong-Woo;Lee, Soo-Choul;Park, Byeong-Gon;Kwon, Il;Sung, Chang-Keun;Park, Wan-Cheol
    • KSBB Journal
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    • v.17 no.1
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    • pp.80-87
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    • 2002
  • Laboratory scale aerobicfanaerobic biofilm reactor was used for simultaneous and stable removal of organics, N and P components to investigate optimum design and operation parameters and to analyze the microbial distribution and consortium structure of nitrification and denitrification bacteria in aerobic and anaerobic biofilm systems. The biofilm reactor was successfully operated for 143 days to show $COD_{cr},\;BOD_5$, SS removal efficiencies of 88, 88, and 97%, respectively. During the experiment period, almost complete nitrification efficiency of 96% was achieved. Denitrification efficiency was about 45% without addition of any external carbon sources. In case of total phosphorus removal, 74% of the inlet phosphorus was removed. Fluorescence in situ hybridization (FISH) results showed that most of the ammonia oxidizing bacteria in the aerobic nitrification zone was found to be Nitrosomonas species while Nitrospira was the representative nitrite oxidizing bacteria. For the denitrification, Rhodobacter, Rhodovulum, Roseebacter and Paracouus were the dominant denitrification bacteria which was 10 to 20% of the total bacteria in numbers.