• Title/Summary/Keyword: Fluorescence in situ hybridization analysis

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Performance and Microbial Characteristics of Bio-hydrogen Production from Food Waste with Thermal Pre-treatment (음식물류 폐기물의 혐기성 수소 발효 시 열처리에 따른 성능 및 미생물 특성 평가)

  • Lee, Chaeyoung;Choi, Jaemin
    • Transactions of the Korean hydrogen and new energy society
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    • v.25 no.1
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    • pp.11-18
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    • 2014
  • This study was conducted to investigate the effect of thermal pre-treatment on bio-hydrogen from food waste. Two continuous reactors operated and VFAs(volatile fatty acids) production and microbial communities were analyzed. The average hydrogen yield was 0.50 and 0.33mol $H_2/mol$ $hexose_{added}$ in thermally treated food added reactor(R1) and control(R2), respectively. Butyrate concentration was similarly 7,500mg/L in both reactors, but two times higher lactate concentration was observed in R2(3,800mg/L). The results of FISH(fluorescence in situ hybridization) showed that the relative microorganism to hydrogen producing bacteria was 78 and 27% in R1 and R2, respectively.

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.

Diagnostic Value of Fluorescence in Situ Hybridization Assay in Malignant Mesothelioma: A Meta-analysis

  • Wan, Chun;Shen, Yong-Chun;Liu, Meng-Qi;Yang, Ting;Wang, Tao;Chen, Lei;Yi, Qun;Wen, Fu-Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4745-4749
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    • 2012
  • The diagnosis of malignant mesothelioma (MM) remains a clinical challenge and the fluorescence in situ hybridization (FISH) assay has been reported to be one promising tool. The present meta-analysis aimed to establish the overall diagnostic accuracy of FISH for diagnosing MM. After a systematic review of English language studies, the sensitivity, specificity and other measures of accuracy of FISH in the diagnosis of MM were pooled using random-effects models. Summary receiver operating characteristic curves were applied to summarize overall test performance. Nine studies met our inclusion criteria, the pooled sensitivity and specificity for FISH for diagnosing MM being 0.72 (95% CI 0.67-0.76) and 1.00 (95% CI 0.98-1.00), respectively. The positive likelihood ratio was 34.5 (95% CI 14.5-82.10), the negative likelihood ratio was 0.24 (95% CI 0.16-0.36), and the diagnostic odds ratio was 204.9 (95% CI 76.8-546.6), the area under the curve being 0.99. Our data suggest that the FISH assay is likely to be a useful diagnostic tool for confirming MM. However, considering the limited studies and patients included, further large scale studies are needed to confirm these findings.

The first Korean case of a newborn with 3p26 microdeletion and 5q35 microduplication inherited from paternal balanced translocation

  • Jang, Jin A;Sohn, Young Bae;Lee, Jang Hoon;Park, Moon Sung
    • Journal of Genetic Medicine
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    • v.18 no.1
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    • pp.48-54
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    • 2021
  • Genetic imbalances are a major cause of congenital and developmental abnormalities. We report the first case of a 3p26 microdeletion and 5q35.2q35.3 microduplication in a newborn with multiple congenital anomalies evaluated using chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH). The patient was born at 30 weeks and 2 days of gestation with a body weight of 890 g. He had symmetric intrauterine growth restriction, microcephaly, facial dysmorphism (hypertelorism, blepharophimosis, mild low-set ears, high-arched palate, and micrognathia), and right thumb polydactyly. Echocardiography revealed an atrial septal defect and patent ductus arteriosus. Furthermore, CMA revealed a concurrent microdeletion in 3p26 and a microduplication in 5q35.2q35.3. FISH analysis showed that these genetic changes resulted from a translocation mutation between chromosomes 3 and 5. The patient's mother had mild intellectual disability, short stature, and facial dysmorphism, while his father had a normal phenotype. However, parental FISH analysis revealed that the asymptomatic father carried a balanced translocation of chromosomes 3p26 and 5q35. CMA and FISH tests are useful for diagnosing neonates with multiple congenital abnormalities. Further parental genetic investigation and proper genetic counseling are necessary in cases of chromosomal abnormalities inherited from parental balanced translocations.

Highly Sensitive Biological Analysis Using Optical Microfluidic Sensor

  • Lee, Sang-Yeop;Chen, Ling-Xin;Choo, Jae-Bum;Lee, Eun-Kyu;Lee, Sang-Hoon
    • Journal of the Optical Society of Korea
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    • v.10 no.3
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    • pp.130-142
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    • 2006
  • Lab-on-a-chip technology is attracting great interest because the miniaturization of reaction systems offers practical advantages over classical bench-top chemical systems. Rapid mixing of the fluids flowing through a microchannel is very important for various applications of microfluidic systems. In addition, highly sensitive on-chip detection techniques are essential for the in situ monitoring of chemical reactions because the detection volume in a channel is extremely small. Recently, a confocal surface enhanced Raman spectroscopic (SERS) technique, for the highly sensitive biological analysis in a microfluidic sensor, has been developed in our research group. Here, a highly precise quantitative measurement can be obtained if continuous flow and homogeneous mixing condition between analytes and silver nano-colloids are maintained. Recently, we also reported a new analytical method of DNA hybridization involving a PDMS microfluidic sensor using fluorescence energy transfer (FRET). This method overcomes many of the drawbacks of microarray chips, such as long hybridization times and inconvenient immobilization procedures. In this paper, our recent applications of the confocal Raman/fluorescence microscopic technology to a highly sensitive lab-on-a-chip detection will be reviewed.

Quantification of Bacillus Species in a Wastewater Treatment System by the Molecular Analyses

  • Mori Koji;Iriye Ryozo;Hirata Mutsunori;Takamizawa Kazuhiro
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.6
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    • pp.482-489
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    • 2004
  • Bacillus species were observed and quantified by molecular approaches, using the 16S rDNA primers/probes, in a wastewater treatment plant designed for the purpose of stimulating the growth of Bacillus species. The plant has been operating as a test plant since 1997 in the city of Ina, Japan, with excellent treatment performance. Observations by in situ hybridization, using Bacillus-specific probes, indicated that Bacillus strains were inhabited in the plant and their num­bers decreased during the treatment process. Similar results were obtained from a quantitative PCR analysis using a Bacillus-specific primer set, and the amount of DNA originating from various Bacillus species was maximally $1.91%\$ of the total DNA in the wastewater treatment tank. Clone library analysis using the Bacillus-specific primers suggested that, while the population was no­ticeably increased, the phylogenetic diversity of the increasing Bacillus species was very low.

Microbial Community Analysis of 5-Stage Biological Nutrient Removal Process with Step Feed System

  • Park, Jong-Bok;Lee, Han-Woong;Lee, Soo-Youn;Lee, Jung-Ok;Bang, Iel-Soo;Park, Eui-So;Park, Doo-Hyun;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.6
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    • pp.929-935
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    • 2002
  • The 5-stage biological nutrient removal (BNR) process with step feed system showed a very stable organic carbon and nutrient removal efficiency ($87\%\;COD\,;79\%\;nitrogen,\;and\;87\%$ phosphorus) for an operation period of 2 years. In each stage at the pilot plant, microbial communities, which are important in removing nitrogen and phosphorus, were investigated using fluorescence in-situ hybridization (FISH) and 165 rDNA characterization. All tanks of 5-stage sludge had a similar composition of bacterial communities. The totat cell numbers of each reactor were found to be around $2.36-2.83{\times}10^9$ cells/ml. About $56.5-62.0\%$ of total 4,6-diamidino-2-phenylindol (DAPI) cells were hybridized to the bacterial-specific probe EUB388. Members of ${\beta}$-proteobacteria were the most abundant proteobacterial group, accounting for up to $20.6-26.7\%$. The high G+C Gram-positive bacterial group and Cytophaga-Flexibacter cluster counts were also found to be relatively high. The beta subclass proteobacteria did not accumulate a large amount of polyphosphate. The proportion of phosphorus-accumulating organisms (PAOs) in the total population of the sludge was almost $50\%$ in anoxic-1 tank. The high G+C Gram-positive bacteria and Cytophaga-Flexibacter cluster indicate a key role of denitrifying phosphorus-accumulating organisms (dPAOs). Both groups might be correlated with some other subclass of proteobacteria for enhancing nitrogen and phosphorus removal in this process.

Prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus

  • Lee, Dongsook;Park, Heeju;Kwak, Sanha;Lee, Soomin;Go, Sanghee;Park, Sohyun;Jo, Sukyung;Kim, Kichul;Lee, Seunggwan;Hwang, Doyeong
    • Journal of Genetic Medicine
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    • v.13 no.2
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    • pp.95-98
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    • 2016
  • We report the prenatal diagnosis of an unbalanced translocation between chromosome Y and chromosome 15 in a female fetus. Cytogenetic analysis of parental chromosomes revealed that the mother had a normal 46,XX karyotype, whereas the father exhibited a 46,XY,der(15)t(Y;15) karyotype. We performed cytogenetic analysis of the father's family as a result of the father and confirmed the same karyotype in his mother and brother. Fluorescence in situ hybridization and quantitative fluorescent-polymerase chain reaction analysis identified the breakpoint and demonstrated the absence of the SRY gene in female members. Thus, the proband inherited this translocation from the father and grandmother. This makes the prediction of the fetal phenotype possible through assessing the grandmother. Therefore, we suggest that conventional cytogenetic and molecular cytogenetic methods, in combination with family history, provide informative results for prenatal diagnosis and prenatal genetic counseling.

Dynamics of in situ Bacterial Community Structure in the Nak-Dong River (낙동강에서의 세균군집구조의 역동성)

  • Park, Ji-Eun;Yeo, Sang-Min;Lee, Young-Ok
    • Korean Journal of Ecology and Environment
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    • v.37 no.4 s.109
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    • pp.363-367
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    • 2004
  • For comparative analysis of the eubacterial community structure at 8 sampling sites throughout the Nak-Dong River, FISH (fluorescence in situ hybridization) method was employed. The total ratio of each determined eubacterial group such as ${\alpha}\;{\cdot}\;{\beta}\;{\cdot}\;{\gamma}-subclasses$proteobacteria and Cytophaga-Flavobacterium(CF) group to total counts(DAPI) at each site varied 9.3-42.5% with the highest value at uppermost part. And each ratio of determined eubacterial groups reached mostly under 10% except that of CF group (23%) at uppermost part. Furthermore, compared to lower part, upper part represented unexpectedly higher proportions of ${\gamma}-subclass$ proteobacteria comprised almost fast growing bacteria on degradable organics. Also the variations of ammonia-oxidizing bacteria ranged from $2.7{\times}10^4$ to $18.0{\times}10^4$ cells $mL^{-1}$ with the lowest value in lower part and the highest value in mid part whereas those of nitrite-oxidizing bacteria varied 5.2-7.7{\times}10^4$ cells $mL^{-1}$ without noticeable differences throughout the sites. Additionally, the ratio of nitrifying bacteria to total counts ranged from 1.0% to 13.6% with no differences between ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. In conclusion, FISH method introduced in this study for monitoring, normally used for the quantitative analysis of bacteria, provided also good information on their environmental status in the Nak-Dong River.

Characteristics of Microbial Distribution of Nitrifiers and Nitrogen Removal in Membrane Bioreactor by Fluorescence in situ Hybridization (막/생물반응기에서 Fluorescence in situ Hybridization 기법을 이용한 질산화 미생물 분포특성 및 질소제거 연구)

  • Lim Kyoung-Jo;Kim Sun-Hee;Kim Dong-Jin;Cha Gi-Cheol;Yoo Ik-Keun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.257-264
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    • 2006
  • An aerobic submerged membrane bioreactor (MBR) treating ammonium wastewater was studied in respect of nitrification characteristics and distribution of nitrification bacteria over a period of 350 days. MBR was fed with ammonium concentration of 500-1000 mg $NH_4-N/L$ at a nitrogen load of $1-2kg\;N/m^3{\cdot}d$. Overall ammonium oxidation rate increased with dissolved oxygen (DO) concentration, temperature, and sludge retention time (SRT). Under a higher concentration of free ammonia ($NH_3-N$) due to the decrease of ammonium oxidation rate, the nitrite ratio ($NO_2-N/NO_x-N$) in the effluent increased. The sudden collapse of nitrification efficiency accompanied by sludge foaming and the increase of sludge volume index (SVI) was observed unexpectedly during the operation. At the later stage of operation, additional carbon source was fed to the MBR and resulted in twice higher value of SVI and the decrease of ammonium oxidation rate. In fluorescence in situ hybridization (FISH) analysis, genus Nitrosomonas which is specifically hybridized with probe NSM156 was initially the dominant ammonia oxidizing bacteria and the amount of Nitrosospira gradually increased. Nitrospira was the dominant nitrite oxidizing bacteria during whole operational period. Significant amount of Nitrobacter was also detected which might due to the high concentration of nitrite maintained in the reactor.