• Title/Summary/Keyword: Fluorescence in-situ hybridization (FISH)

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Piggery Waste Treatment using Partial Nitritation and Anaerobic Ammonium Oxidation (부분질산화와 혐기성 암모늄산화를 이용한 돈사폐수처리)

  • Hwang, In-Su;Min, Kyung-Sok;Lee, Young-Ok
    • Journal of Korean Society on Water Environment
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    • v.22 no.4
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    • pp.599-604
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    • 2006
  • Nitrogen removal with the combined SHARON (Single reactor system for high ammonium removal over nitrite)ANAMMOX (Anaerobic ammonium oxidation) process using the effluent of ADEPT (Anaerobic digestion elutriated phased treatment) slurry reactor with very low C/N ratio for piggery waste treatment was investigated. For the preceding SHARON reactor, ammonium nitrogen loading and removal rate were $0.97kg\;NH_4-N/m^3_{reactor}/day$ and $0.68kg\;NH_4-N/m^3_{reactor}/day$ respectively. In steady state, bicarbonate alkalinity consumption for ammonium nitrogen converted to $NO_2-N$ or $NO_3-N$ was 8.4 gram per gram ammonium nitrogen. The successive ANAMMOX reactor was fed with the effluent from SHARON reactor. The loading and removal rate of the soluble nitrogen defined as the sum total of $NH_4-N$, $NO_2-N$ and $NO_3-N$ in ANAMMOX reactor were $1.36kg\;soluble\;N/m^3_{reactor}/day$ and $0.7kg\;soluble\;N/m^3_{reactor}/day$, respectively. The average $NO_2-N/NH_4-N$ removal ratio by ANAMMOX was 2.41. Fluorescence in situ hybridization (FISH) analysis verified that Candidatus Kuenenia stuttgartiensis were dominate, which means that they played an important role of nitrogen removal in ANAMMOX reactor.

Comparative Cytogenetic Characteristics and Physical Mapping of the 17S and 5S Ribosomal DNAs between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

  • Bang, Kyong-Hwan;Koo, Dal-Hoe;Kim, Hong-Sig;Song, Beom-Heon;Cho, Yong-Gu;Cho, Joon-Hyeong;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.4
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    • pp.311-315
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    • 2003
  • This study was carried out to compare chromosomal characteristics between Atractylodes japonica and A macrocephala. Cytogenetic analysis was conducted based on karyotype analysis and physical mapping using fluorescence in situ hybridization. As a result of karyotype analysis by feulgen staining, somatic chromosome numbers of A. japonica and A. macrocephala were 2n=24. The length. of the mitotic metaphase chromosomes of A. japonica ranged from $0.70\;to\;1.60{\mu}m$ with a total length. of $12.11{\mu}m$ and the homologous chromosome complement comprised six metacentrics, five submetacentrics and one subtelocentrics. On the other hand, the length of the mitotic metaphase chromosomes of A. macrocephala ranged from $0.90\;to\;2.35{\mu}m$ with a total length of $16.58{\mu}m$ and the homologous chromosome complement comprised seven metacentrics and five submetacentrics. The total length of A. japonica chromosomes was shorter than that of A. macrocephala, but A. japonica had one subtelocentrics (chromosomes 4) different from A. macrocepha1a. chromosomes. The F1SH technique using 17S and 5S rDNA was applied to metaphase chromosomes. The signals for 17S rDNA were detected on the telomeric regions of chromosomes 4 and 5 in both A japonica and A. macrocephala. The 5S rDNA signal was found in the short arm of chromosome 1.

The spectrum of 5p deletion in Korean 20 patients with Cri du chat syndrome (한국인 묘성증후군 20명 환자에서의 5p 결실 양상 분석)

  • Park, Sang-Jin;Kim, Sook-Ryung;Baek, Kum-Nyeo;Yoon, Joon-No;Jeong, Eun-Jeong;Kown, Ji-Eun;Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.4 no.2
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    • pp.133-141
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    • 2007
  • Purpose : Cri-du-Chat syndrome (CdCs) is a rare but clinically recongnizable condition with an estimated incidence of 1:50,000 live births. The clinical characteristics of the syndrome include severe psychomotor and mental retardation, microcephaly, hypertelorism, hypotonia, and slow growth. Also the size of the chromosome 5p deletion ranges were known from the region 5p13 to the terminal region. In this study, we report the spectrum of 5p deletion in Korean 20 pts. with CdCs and genotype-phenotype associations in CdCs. Methods : In order to delineate genotype-phenotype correlation, molecular cytogenetic studies including GTG banding and clinical characterization were performed on Korean 20 pts with CdCs including parents. CGH array and Fluorescence in situ hybridization (FISH) analysis were used to confirm a terminal deletion karyotype and map more precisely the location of the deletion breakpoint. Results : Molecular analysis of the spectrum of 5p deletion revealed 9 pts (45%) with a del (5)(p14), 7 pts. (35%) a del (5)(p13), 3 pts. (15%) a del (5)(p15.1) and 1 pt. (5%) a del (5)(p15.2) in 20 pts with CdCs. 4(20%)pts were identified to have additional chromosome abnormalites of deficiency and duplication involving chromosomes of 6, 8, 18, & 22. Parental study identified 3 familial case (2 paternal and 1 maternal origin) showing parents being a balanced translocation carrier. And the comparison study of the deletion break points among these 20 pts. with their phenotype has showed the varying clinical pheno-types in the CdCs critical region. Conclusion : The characterization of 5p deletion including parental study may help to delineate the genotypephenotype correlation in CdCs. Also these molecular cytogenetic analyses will be able to offer better information for accurate genetic diagnosis in CdCs and further make possible useful genetic counseling in pts. and family.

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The Amount of Telomeric DNA and Telomerase Activity on Cattle Cells (소의 생리적 특성에따름 세포내 텔로미어 함량과 텔로머레이스 활성도 분석)

  • Choi, Duk-Soon;Cho, Chang-Yeon;Sohn, Sea-Hwan
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.445-456
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    • 2008
  • Telomeres consist of TTAGGG tandem repeated DNA sequences with specific proteins and locate at chromosome ends. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Telomerase is a ribonucleoprotein which has a template for the synthesis of telomeric DNA. This study was carried out to analyze the amount of telomeric DNA and telomerase activity in cattle cells. Analysis of the quantity of telomere in lymphocytes was done at different ages, sex and among Korean cattle and Holstein breeds. The telomerase activity was also analyzed in liver, brain, heart, kidney, and testis tissues of fetal calf and of 18 month old cattle. The amount of telomeres in lymphocytes and other tissue cells was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) technique using a telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol assay (TRAP). The amount of telomeric DNA on the lymphocytes during the whole life span was decreased along with age. Quantity of telomeres in Korean cattle was significantly higher than that in Holstein breed. The amount of telomeric DNA in males was significantly higher than that in females. Telomerase activity was up-regulated in most bovine tissues during fetal stage, but was down-regulated in most tissues at mature 18 month age except the testis cells. This study indicates that the amount of telomeres and telomerase activity of cells can be used as an age marker or/and a physiological marker of cattle.

A refined Panax ginseng karyotype based on an ultra-high copy 167-bp tandem repeat and ribosomal DNAs

  • Waminal, Nomar Espinosa;Choi, Hong-Il;Kim, Nam-Hoon;Jang, Woojong;Lee, Junki;Park, Jee Young;Kim, Hyun Hee;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.469-476
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    • 2017
  • Background: Panax ginseng Meyer (Asian ginseng) has a large nuclear genome size of > 3.5 Gbp in haploid genome equivalent of 24 chromosomes. Tandem repeats (TRs) occupy significant portions of the genome in many plants and are often found in specific genomic loci, making them a valuable molecular cytogenetic tool in discriminating chromosomes. In an effort to understand the P. ginseng genome structure, we characterized an ultrahigh copy 167-bp TR (Pg167TR) and explored its chromosomal distribution as well as its utility for chromosome identification. Methods: Polymerase chain reaction amplicons of Pg167TR were labeled, along with 5S and 45S rDNA amplicons, using a direct nick-translation method. Direct fluorescence in situ hybridization (FISH) was used to analyze the chromosomal distribution of Pg167TR. Results: Recently, we reported a method of karyotyping the 24 chromosome pairs of P. ginseng using rDNA and DAPI (4',6-diamidino-2-phenylindole) bands. Here, a unique distribution of Pg167TR in all 24 P. ginseng chromosomes was observed, allowing easy identification of individual homologous chromosomes. Additionally, direct labeling of 5S and 45S rDNA probes allowed the identification of two additional 5S rDNA loci not previously reported, enabling the refinement of the P. ginseng karyotype. Conclusion: Identification of individual P. ginseng chromosomes was achieved using Pg167TR-FISH. Chromosome identification is important in understanding the P. ginseng genome structure, and our method will be useful for future integration of genetic linkage maps and genome scaffold anchoring. Additionally, it is a good tool for comparative studies with related species in efforts to understand the evolution of P. ginseng.

Telomeric Dynamics and Telomerase Activity in Early Bovine Embryos (소의 초기 배 발생단계별 Telomeric DNA 함량 및 Telomerase Activity 분석)

  • Jung, Yei-Hwa;Lee, Soo-Hee;Cho, Sang-Rae;Kong, Il-Keun;Cho, Jae-Dong;Sohn, Sea-Hwan
    • Clinical and Experimental Reproductive Medicine
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    • v.36 no.2
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    • pp.101-109
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    • 2009
  • Objective: This study was carried out to analyze the amount of telomeric DNA and telomerase activity in early bovine embryos. Methods: The amount of telomeric DNA in early bovine embryos at the 8 cell, morula and blastocyst stages was analyzed by Quantitative Fluorescence In Situ Hybridization (Q-FISH) technique using a bovine telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol (TRAP assay). Results: The relative amount of telomeric DNA in early bovine embryos was gradually increased from 8 cell to blastocyst stage. It was not significantly associated with the grade of embryo quality. While telomerase activity was detected in the early bovine embryos at these stages, it significantly increased at morula stage and showed maximum activity at the blastocyst stage. Conclusion: The amount of telomeric DNA and the telomerase activity of bovine embryos increase during the progression of early embryogenesis, suggesting a positive correlation between telomeric DNA and telomerase activity. The telomerase activity seems to increase to maintain the levels of telomeric DNA through embryo development which are required for extensive cell division.

UNDERSTANDING OF EPIGENETICS AND DNA METHYLATION (인간 게놈의 Copy Number Variation과 유전자 질환)

  • Oh, Jung-Hwan;Nishimura, Ichiro
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.30 no.2
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    • pp.205-212
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    • 2008
  • Genetic variation in the human genome occurs on various levels; from the single nucleotide polymorphism to large, microscopically visible chromosome anomalies. It can be present in many forms, including variable number of tandem repeat (VNTRs; e.g., mini- and microsatellites), presence/absence of transposable elements (e.g., Alu elements), single nucleotide polymorphisms, and structural alterations (e.g., copy number variation, segmental duplication, inversion, translocation). Until recently SNPs were thought to be the main source of genetic and phenotypic human variation. However, the use of methods such as array comparative genomic hybridization (array CGH) and fluorescence in situ hybridization (FISH) have revealed the presence of copy number variations(CNVs) ranging from kilobases (kb) to megabases (Mb) in the human genome. There is great interest in the possibility that CNVs playa role in the etiology of common disease such as HIV-1/AIDS, diabetes, autoimmune disease, heart disease and cancer. The discovery of widespread copy number variation in human provides insights into genetic variability among populations and provides a foundation for studies of the contribution of CNVs to evolution and disease.

Effect of Maternal Age on Chromosome Aberrations and Telomere Quantity in Chick Embryos (닭의 모체 연령에 따른 생산 배아의 염색체 이상 빈도 및 텔로미어 함량 분석)

  • Lee, Soo-Hee;Subramani, Vinod K.;Sohn, Sea-Hwan
    • Korean Journal of Poultry Science
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    • v.36 no.4
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    • pp.293-300
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    • 2009
  • The rate of fetus with abnormal chromosomes increase with maternal age. Nondisjunction of aging oocyte chromosome is a major reason for the increased rate of abnormalities. Telomeres are the ends of eukaryotic chromosome, which are essential for chromosome stability and are related in cell senescence. This study was carried out to analyze the chromosome aberration rate and amount of telomeric DNA in chick embryo along with maternal age. Fertilized eggs and blood were sampled from White Leghorn layers starting at 20 weeks through to 70 weeks age at 10 weeks interval. Chromosome aberration rate was analyzed by karyotyping. The amounts of telomeric DNA in embryonic cells and lymphocytes were quantified by Quantitative Fluorescence in situ Hybridization method. The chromosome aberration rate in chick embryos significantly differed with maternal age. The chromosome aberration rate increased at early laying period and beyond 70 weeks of maternal age. Therefore, chromosome aberration rate was affected by maternal age due to ovulated oocytes state. However, the amount of telomeric DNA on embryonic cells did not differ significantly with maternal age. Thus, maternal age does not affects telomere quantity in their embryos due to cellular reprograming at early embryonic stage after fertilization.

Prenatal Diagnosis of a Satellited Chromosome 8p Results from a de novo Cryptic Translocation between Chromosomes 8 and 22 (산전 진단에서 관찰된 8번과 22번 염색체 사이의 미세 전좌에 의한 8번 염색체 단완 위성체)

  • Oh, Ah-Rum;Lee, Bom-Yee;Choi, Ene-Yuong;Ryu, Hyun-Mee;Lee, Seung-Jae;Jung, Ji-Ye;Park, So-Yeon
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.135-138
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    • 2011
  • The authors of the present study report the prenatal detection of a chromosomal abnormality with additional satellites on the distal short arm of chromosome 8. A 35-year-old woman was referred for amniocentesis because of her advanced maternal age and positive result for maternal serum screening test. Cytogenetic analysis of cultured amniocytes showed a satellite 8p chromosome. The satellite 8p chromosome was positive for nucleolus organizer region (NOR) staining. The parents' karyotypes were normal. Fluorescence in situ hybridization (FISH) study for metaphases of fetal amniocytes revealed a cryptic translocation of chromosomes 8p and 22p. The fetal karyotype was described as 46,XY,8ps.ish t(8;22)(p23.3;p11.2) (D8S504-;D8S504+)dn. The parents decided to continue the pregnancy and a phenotypically normal boy was born at 38 weeks of gestation. In case of de novo terminal NORs detected prenatally, more accurate cytogenetic and molecular analysis should be performed in order to rule out cryptic chromosomal rearrangement among other chromosomes.

Diagnosis and Monitoring of Chronic Myeloid Leukemia: Chiang Mai University Experience

  • Tantiworawit, Adisak;Kongjarern, Supanat;Rattarittamrong, Ekarat;Lekawanvijit, Suree;Bumroongkit, Kanokkan;Boonma, Nonglak;Rattanathammethee, Thanawat;Hantrakool, Sasinee;Chai-Adisaksopha, Chatree;Norasetthada, Lalita
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.2159-2164
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    • 2016
  • Background: A diagnosis of chronic myeloid leukemia (CML) is made on discovery of the presence of a Philadelphia (Ph) chromosome. The success of the treatment of this form of leukemia with tyrosine kinase inhibitor (TKI) is monitored by reduction of the Ph chromosome. Objective: To compare the role of conventional cytogenetic (CC) methods with a real time quantitative polymerase chain reaction (RQ-PCR) and fluorescence in situ hybridization (FISH) for diagnosis and treatment monitoring of CML patients. The secondary outcome was to analyze the treatment responses to TKI in CML patients. Materials and Methods: This was a retrospective study of CML patients who attended the Hematology clinic at Chiang Mai University Hospital from 2005-2010. Medical records were reviewed for demographic data, risk score, treatment response and the results of CC methods, FISH and RQ-PCR. Results: One hundred and twenty three cases were included in the study, 57.7% of whom were male with a mean age of 46.9 years. Most of the patients registered as intermediate to high risk on the Sokal score. At diagnosis, 121 patients were tested using the CC method and 118 (95.9%) were identified as positive. Five patients failed to be diagnosed by CC methods but were positive for BCR-ABL1 using the FISH method. Imatinib was the first-line treatment used in 120 patients (97.6%). In most patients (108 out of 122, 88.5%), a complete cytogenetic response (CCyR) was achieved after TKI therapy and in 86 patients (70.5%) CCyR was achieved long term by the CC method. Five out of the 35 analyzed patients in which CCyR was achieved by the CC method had a positive FISH result. Out of the 76 patients in which CCyR was achieved, RQ-PCR classified patients to only CCyR in 17 patients (22.4%) with a deeper major molecular response (MMR) in 4 patients (5.3%) and complete molecular response (CMR) in 55 patients (72.4%). In the case of initial therapy, CCyR was achieved in 95 patients (79.1%) who received imatinib and in both patients who received dasatinib (100%). For the second line treatment, nilotinib were used in 30 patients and in 19 of them (63.3%) CCyR was achieved. In half of the 6 patients (50%) who received dasatinib as second line or third line treatment CCyR was also achieved. Conclusions: CML patients had a good response to TKI treatment. FISH could be useful for diagnosis in cases where CC analysis failed to detect the Ph chromosome. RQ-PCR was helpful in detecting any residual disease and determining the depth of the treatment response at levels greater than the CC methods.