• Title/Summary/Keyword: Fluorescence In Situ Hybridization

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Reanalysis of discarded blastocysts for autosomal aneuploidy after sex selection in cleavage-stage embryos

  • Ebrahimian, Neda;Montazeri, Fatemeh;Sadeghi, Mohammad Reza;Kalantar, Seyed Mehdi;Gilany, Kambiz;Khalili, Mohannad Ali
    • Clinical and Experimental Reproductive Medicine
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    • v.47 no.4
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    • pp.293-299
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    • 2020
  • Objective: The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. Methods: Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. Results: Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). Conclusion: The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.

Extraosseous Ewing's Sarcoma Presented as a Rectal Subepithelial Tumor: Radiological and Pathological Features

  • Bae, Heejin;Chung, Taek;Park, Mi-Suk;Kim, Myeong-Jin;Lim, Joon Seok;Kim, Honsoul
    • Investigative Magnetic Resonance Imaging
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    • v.21 no.1
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    • pp.51-55
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    • 2017
  • Purpose: Extraosseous Ewing's sarcoma (EOE) of the rectum is extremely rare: only three cases have been reported in the literature and none of these reports described their imaging findings in detail. Herein, we describe the tumor imaging and pathological features in detail. Materials and Methods: We report a case of rectal EOE in a 72-year-old female who received local excision and was provisionally diagnosed with a rectal submucosal spindle cell tumor. We used immunohistochemistry, histopathology, and fluorescence in situ hybridization to characterize the tumor and provide a definitive diagnosis of EOE. Results: MRI revealed a well-demarcated submucosal tumor with heterogeneous enhancement and hemorrhagic foci in rectum. EOE was diagnosed by positive staining of tumor cells for CD99 and Fli-1 by immunohistochemistry and the presence of the EWSR1 gene translocation by fluorescence in situ hybridization. Although the patient underwent radiation treatment and surgery, the tumor recurred after 4 months as revealed by computed tomography and magnetic resonance imaging. Conclusion: Rectal EOE may present as a rectal submucosal tumor. The understanding of imaging and histological characteristics of this tumor are critical for accurate diagnosis and appropriate aggressive treatment.

A Case of Successful Pregnancy in Patient with Recurrent Spontaneous Abortion by Preimplantation Genetic Diagnosis Following IVF-ET (염색체 이상에 의한 반복 유산 환자에서 체외수정시술 및 착상전 유전진단을 통한 임신 성공 1례)

  • Jeong, Jin-Seok;Yeon, Gyu-Sun;Chae, Hee-Dong;Cheon, Yong-Pil;Kim, Chung-Hoon;Kang, Byung-Moon;Chang, Yoon-Seok;Mok, Jung-Eun
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.135-140
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    • 1998
  • It was reported that the etiologies of recurrent spontaneous abortion are immunologic factors, endocrinologic problems, anatomical abnormalities, genetic abnormalities, infection, and unexplained factors. Among those etiologic factors, genetic abnormalities occur in about 5% of the couples who experience recurrent spontaneous abortions, and most common parental chromosomal abnormality contributing to recurrent abortion is balanced translocation. The advent of in vitro fertilization (IVF), the development of skills associated with the handling of human embryo, and an explosion of knowledge in molecular biology have opened the possibility of early diagnosis of genetic disease in preimplantation embryos. Therefore preimplantation genetic diagnosis (PGD) is indicated for couples, infertile or not, at risk of transmitting a genetic disease. A case of successful pregnancy and term delivery by PGD using fluorescence in situ hybridization (FISH) technique in patient with recurrent spontaneous abortion due to balanced translocation is presented with brief review of literatures.

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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.

Effects of Nitrifying Bacterial Communities with Different HRTs and Backwashing Periods in Modified BAF Process (Modified BAF 공정에서 HRT 및 역세주기가 질산화 미생물의 군집에 미치는 영향)

  • Jung, Chul-Soo;Park, Jeung-Jin;Ju, Dong-Jin;Kwon, Soo-Youn;Choi, Won-Seok;Byun, Im-Gyu;Park, Tae-Joo
    • Journal of Korean Society on Water Environment
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    • v.23 no.6
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    • pp.920-926
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    • 2007
  • The upflow Biobead$^{(R)}$ process, one of biological aerated filters (BAF), which was used commercially, invented for removal of organic materials and nitrification. This process was modified to enhance the ability of denitrification through the induction of pre-anoxic tank. In this research, we investigated the effects of hydraulic retention time (HRT) and backwashing period in aerobic tank. The characteristics of nitrifying bacteria, which are composed of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB), also investigated using fluorescence in situ hybridization (FISH). Even though the HRT was shortened, the efficiency of nitrification was not decreased when the organic loading rate and ammonium-nitrogen loading rate were $2.10kg/m^3/day$ and $0.25kg/m^3/day$, respectively. And then the distribution ratios of AOB and NOB showed the similar patterns. However, when the backwashing period was lengthened from 12 hours to 24 hours in aerobic 1 tank, the nitrification efficiency was decreased to 63.9% from 89.2%. The results of FISH explained that this decrease of nitrification efficiency was caused by the decrease of distribution ratio of AOB in aerobic 1 tank. The nitrification efficiencies of aerobic 1 and aerobic 2 tank were increased when the backwashing period was lengthened because of relative high distribution ratios of nitrifying bacteria.

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.

The Effect of Fixed Media and Recycling Ratio on Nutrients Removal in a Pilot-Scale Wastewater Treatment Unit (고정식 담체 유무와 반송비에 따른 소규모 하수처리 시스템 내 영양염류 제거 특성)

  • Hwang, Jae-Hoon;Cho, Dong-Wan;Kim, Chung-Hwan;Jeon, Byong-Hun
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.6
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    • pp.449-455
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    • 2013
  • The effect of recycling ratio and fixed media on nitrate and phosphate removal was investigated in a pilot-scale wastewater treatment unit using synthetic wastewater. Addition of fixed media increased nitrate removal from 45 to 58% while no noticeable change was observed for Chemical Oxygen Demand (COD) and phosphate removal (<5%). Nitrate removal efficiency also enhanced (Ca 7%) when the influent wastewater flow was doubled (2Q), however phosphate removal was decreased from 40.9 to 26.6% with the increasing recycling rate. The attached biomass analysis showed the presence of bacteria (73.4 $mg/cm^2$) on the surface of added media in anoxic reactor. Pseudomonas aeruginosa a common denitrifying bacterium dominated the bacterial growth (58%) in the anoxic reactor which was determined using Fluorescence In Situ Hybridization (FISH) analysis.

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.

Karyotype Analysis and Physical Mapping of rDNAs in Bupleurum longeradiatum (개시호 (Bupleurum longeradiatum)의 핵형분석과 rDNAs의 Physical Mapping)

  • Koo, Dal-Hoe;Seong, Nak-Sul;Seong, Jong-Suk;Bang, Kyong-Hwan;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.402-407
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    • 2003
  • Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in Bupleurum longeradiatum. Somatic metaphase chromosome number was 2n=12. Karyotype was composed of three pairs of metacentrics (No.3, 4 and 6) and three pairs of submetacentrics (No. 1, 2 and 5). The length of somatic prometaphase chromosomes ranges from 2.55 to $5.05{\mu}m$ with total length of $18.15\;{\mu}m$. In FISH experiment, one pair of 5S rDNA signals was detected on the pericentromeric region of chromosome 4 and one pair of 45S rDNA signals was detected on the telomeric region of chromosome 2.

Telomere의 양적 분석을 이용한 닭의 bio-marker개발

  • 조은정;최철환;전익수;박철;손시환
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2004.11a
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    • pp.13-15
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    • 2004
  • Telomeres are the end of chromosomes and consist of a tandem repeat sequence of (TTAGGG)n and associated proteins. Telomeres are essential for chromosome stability and are related with cell senescence and apoptosis. This study was carried out to analyze the amount of telomeric DNA of chicken lymphocytes, which is to considered as bio-marker. The amount of telomeric DNA of lymphocytes in Korean Native Chicken and White Leghorn was analyzed by quantitative-fluorescence in situ hybridization (Q-FISH) technique using the chicken telomeric DNA probe. Telomere quantifies were compared among breeds, ages and sex, and the relationship between the amount of telomeres and their productive trait was also analyzed. Comparing the amount of telomeric DNA on lymphocytes during growing period, the amount of telomeres was gradually decreased as growing older. The telomere quantity was also significantly different in breeds and sex. Estimating correlation coefficient, the amount of telomeres was positively correlated to sexual maturity and body weight but negatively correlated to hen day egg production and egg weight. These results implicate the telomere quantity is considered as an individual bio-marker.

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