• 제목/요약/키워드: Fluorescent in situ hybridization

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The Effects of Human Adipose Tissue-derived Stem Cells on Degenerative Change of Knee in Rabbit Model (가토 모델에서 인체지방유래 줄기세포가 슬관절의 퇴행성 변화에 미치는 영향)

  • Jeong, Ki-Hwan;Kim, Seok-Kwun;Jeong, Jae-Oo;Heo, Jeong;Kwon, Yong-Seok;Lee, Keun-Cheol
    • Archives of Plastic Surgery
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    • v.35 no.6
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    • pp.637-644
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    • 2008
  • Purpose: The survival of bone marrow derived stem cell was reported several times. But the survival of adipose tissue derived stem cells(hASCs) was not mentioned on. We studied the adipose tissue derived stem cell's survival and effect on articular cartilage in rabbits. Methods: Osteoarthritis was induced in twenty New Zealand white rabbits by intraarticular injection of monosodium iodoacetate(MIA). After four weeks, hASCs were also injected into the knee joints space without any vehicle, but the control group received phosphate buffered saline only. The histologic grade of articular cartilage was measured in 4 and 8 weeks after the transplantation of hASC and the viability of injected stem cells measured by Fluorescent in situ Hybridization (FISH) examination. Results: After 4 and 8 weeks from hASCs transplantation, histologic grade was not significantly difference between two groups(p>0.05), and the Y chromosome of the transplanted hASCs was not detected in articular cartilage. Conclusion: We found that direct injection of hASC in joint space didn't work on damaged articular cartilage repair.

Genetic overgrowth syndrome: A single center's experience

  • Cheon, Chong Kun;Kim, Yoo-Mi;Yoon, Ju Young;Kim, Young A
    • Journal of Genetic Medicine
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    • v.15 no.2
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    • pp.64-71
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    • 2018
  • Purpose: Overgrowth syndromes are conditions that involve generalized or localized areas of excess growth. In this study, the clinical, molecular, and genetic characteristics of Korean patients with overgrowth syndrome were analyzed. Materials and Methods: We recruited 13 patients who presented with overgrowth syndrome. All patients fulfilled inclusion criteria of overgrowth syndrome. Analysis of the clinical and molecular investigations of patients with overgrowth syndrome was performed retrospectively. Results: Among the 13 patients with overgrowth syndrome, 9 patients (69.2%) were found to have molecular and genetic causes. Among the seven patients with Sotos syndrome (SS), two had a 5q35microdeletion that was confirmed by fluorescent in situ hybridization. In two patients with SS, intragenic mutations including a novel mutation, c.5993T>A (p.M1998L), were found by Sanger sequencing. One patient had one copy deletion of NDS1 gene which was confirmed by multiplex ligation-dependent probe amplification. Among five patients with Beckwith-Wiedemann syndrome, three had aberrant imprinting control regions; 2 hypermethylation of the differentially methylated region of H19, 1 hypomethylation of the differentially methylated region of Kv. In one patient displaying overlapping clinical features of SS, a de novo heterozygous deletion in the chromosomal region 7q22.1-22.3 was found by single nucleotide polymorphism-based microarray. Conclusion: Considering high detection rate of molecular and genetic abnormalities in this study, rigorous investigations of overgrowth syndrome may be an important tool for the early diagnosis and genetic counseling. A detailed molecular analysis of the rearranged regions may supply the clues for the identification of genes involved in growth regulation.

Inhibitory Effect of Aged Petroleum Hydrocarbons on the Survival of Inoculated Microorganism in a Crude-Oil-Contaminated Site

  • Kang, Yoon-Suk;Park, Youn-Jong;Jung, Jae-Joon;Park, Woo-Jun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.12
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    • pp.1672-1678
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    • 2009
  • We studied the effects of aged total petroleum hydrocarbons (aged TPH) on the survival of allochthonous diesel-degrading Rhodococcus sp. strain YS-7 in both laboratory and field investigations. The aged TPH extracted from a crude-oil-contaminated site were fractionized by thin-layer chromatography/flame ionization detection (TLC/FID). The three fractions identified were saturated aliphatic (SA), aromatic hydrocarbon (AH), and asphaltene-resin (AR). The ratio and composition of the separated fractions in the aged TPH were quite different from the crude-oil fractions. In the aged TPH, the SA and AH fractions were reduced and the AR fraction was dramatically increased compared with crude oil. The SA and AH fractions (2 mg/l each) of the aged TPH inhibited the growth of strain YS-7. Unexpectedly, the AR fraction had no effect on the survival of strain YS-7. However, crude oil (1,000 mg/l) did not inhibit the growth of strain YS-7. When strain YS-7 was inoculated into an aged crude-oil-contaminated field and its presence was monitored by fluorescent in situ hybridization (FISH), we discovered that it had disappeared on 36 days after the inoculation. For the first time, this study has demonstrated that the SA and AH fractions in aged TPH are more toxic to an allochthonous diesel-degrading strain than the AR fraction.

Dynamics Behavior of Phage-Host System Related to Microlunatus phosphovorus in Activated Sludge with Host Inoculation

  • Lee, Sang-Hyon;Otawa, Kenichi;Onuki, Motoharu;Satoh, Hiroyasu;Mino, Takashi
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1518-1522
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    • 2006
  • In the present study, it was observed how the phage-host system that is naturally reproduced in activated sludge is affected by the host inoculation. The system of Microlunatus phosphovorus and its phages was selected as the phage-host system native to an activated sludge system operated for 19 days under sequencing anaerobic-aerobic conditions with glutamate as the main carbon source. The phage-host system related to M. phosphovorus was monitored by plaque assay for the phages and by fluorescent in situ hybridization (FISH) for the bacterial host. In addition, the whole phage structure was also monitored by pulsed-field gel electrophoresis (PFGE). During the first 9 days, the phage-host system was more or less steady at approx. 9% (FISH/ DAPI) for M. phosphovorus and approx. 10,000 PFU/ml for its lytic phages. Microlunatus phosphovorus JCM9379 was inoculated into the activated sludge on day 10. Right after the inoculation, M. phosphovorus was approx. 24% (FISH/DAPI) whereas its lytic phages dropped down to approx. 500 PFU/ ml. After the host inoculation (within 9 days), however, the phage-host system eventually reverted to its original level in each population. On the other hand, the whole phage structure was not significantly changed by M. phosphovorus inoculation but stable throughout the process operation. Only the minor change that four phage groups gradually became abundant after the host inoculation was observed.

Enrichment of Ammonia-Oxidizing Bacteria for Efficient Nitrification of Wastewater

  • KIM WON-KYOUNG;CUI RONG;JAHNG DEOKJIN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.772-779
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    • 2005
  • Ammonia-oxidizing bacteria (AOB) were enriched by repeating fed-batch cultivations in an AOB-selective medium of activated sludges from a domestic wastewater treatment plant. Enriched culture showed strong capabilities of ammonia oxidation [0.810 mg $NH_4^+$-N/mg mixed liquor suspended solids (MLSS)$\cdot$day] as well as $NO_x^-$-N production (0.617 mg $NO_x^-$-N/ mg MLSS$\cdot$day). Degree of enrichment was examined through fluorescent in situ hybridization (FISH) analyses using an AOB-specific Cy3-labeled oligonucleotide probe (NSOl90) and terminal-restriction fragment length polymorphism (T-RFLP) analyses. FISH analyses confirmed that the fraction of AOB among 4',6-diamidino-2-phenylindole (DAPI)-stained cells increased from about less than $0.001\%$ to approximately $42\%$ after enrichment of AOB, and T-RFLP analyses showed that bacterial community became simpler as enrichment was continued. When the enriched culture of AOB was added (150 mg/l as dry suspended solid) to the normal activated sludge (3,000 mg/l as dry suspended solid), nitrification efficiencies were improved from 0.020 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.041 mg $NO_x^-$-N/mg MLSS$\cdot$day in a synthetic wastewater and also from 0.0007 mg $NO_x^-$-N/mg MLSS$\cdot$day to 0.0918 mg $NO_x^-$-N/mg MLSS$\cdot$day in a real domestic wastewater. Therefore, it is expected that this enrichment method could be used for improving efficiency of nitrification in wastewater treatment plants.

Clinicopathology Figures and Long-term Effects of Tamoxifen Plus Radiation on Survival of Women with Invasive Ductal Carcinoma and Triple Negative Breast Cancer

  • Payandeh, Mehrdad;Sadeghi, Masoud;Sadeghi, Edris;Aeinfar, Mehrnoush
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.12
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    • pp.4863-4867
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    • 2015
  • Background: Triple negative breast cancer (TNBC), characterized as estrogen receptor (ER), progesterone receptor (PR) and human epidermal growth factor receptor 2 Her2 negative and accounting for 10-17% of all breast carcinomas, is only partially responsive to chemotherapy and suffers from a lack of clinically established targeted therapies. The aim of the current study was to evaluate the patterns of treatment and clinicopathology figures in Kurdish patients with triple-negative breast cancer, and to compare these to other reports. Materials and Methods: Between 2001 and 2014, 950 breast cancer patients were referred to our clinic. There were 74 female patients with TNBC, including 70 patients was invasive ductal carcinoma entered into our study. ER and PR positivity was defined as positive immunohistochemical staining in more than 10% of tumor cells. Immunohistochemistry assay with anti-HER2 antibodies was used to identify HER negative (0 and 1+) or positive (2+ and 3+). HER2 gene amplification was determined by fluorescent in situ hybridization (FISH). Overall survival (OS) was plotted with GraphPad Prism 5 Software using Kaplan-Meier and log-rank tests for comparison of results. Results: The mean age in the first diagnosis for 70 patients with triple TNBC and invasive ductal carcinoma was 49.6 years that range of age was 27-82 years. All of the patients were female. Of 70 patients, 23 patients had metastasis. Thirty-two patients (45.7%) were treated with tamoxifen and 39 (55.7%) with radiotherapy. Three-year, 5-year and 10-year OS rates for all patients were 82%, 72% and 64%, respectively. Conclusions: The OS in our West Iran TNBC patients is less than reported elsewhere. However, treatment with combination of tamoxifen plus radiation increases the OS and reduces the mortality rate.

Molecular probe for identification of cysts of resting cyst of PSP-producer Alexandrium tamarense (Dinophyceae) (분자생물학적 방법을 이용하여 마비성 패류 독소를 생산하는 알렉산드륨 타마렌스 시스트 탐색)

  • Cho, Eun-Seob
    • Journal of Life Science
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    • v.13 no.2
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    • pp.163-167
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    • 2003
  • Identification of species within the toxin-producing genus Alexandrium is vital for biotoxin monitoring and mitigation decisions regarding shellfish industry. In particular, the discrimination of resting cysts of only A. tamarense from that of Alexandrium spp. is considerable important to fundamentally monitor and predict this species before vegetative cells occur in the nature. Fluorescent cTAM-F1 DNA probe was responsible to not only binding the activity of the vegetative cells in A. tamarense, but also to the resting cysts, which was treated with methanol after fixation and stained by primuline on the surface The location of fluorescence in cultured vegetative cells and resting cysts was almost at tile bottom of the nucleus. The optimal incubation temperature and time using in situ hybridization were 50-$54^{\circ}C$ and 40-60 min, respectively, to penetrate the DNA probe into cell.

Molecular Analysis of the Y Chromosome in a 46,XY Female Phenotype

  • Kim, Jin-Woo;Kim, Tae-Jin;Park, So-Yeon;Nam, Sung-A;Jun, Jong-Young
    • Journal of Genetic Medicine
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    • v.3 no.1
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    • pp.5-10
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    • 1999
  • This is a case report of 46,XY female phenotype (46,XY karyotype, no pubic hair, blind vagina and absence of uterus)in an 18-year-old patient. To confirm whether a Y chromosome has a structural abnormality, fluorescent in situ hybridization (FISH) with the chromosome X/Y cocktail probe was simultaneously performed, and the six loci [PABY, RPS4Y(sy16, sy17), ZFY, DYS14] on the short arm, one locus (DYZ3) on the centromere and one locus (DYZ1) on the long arm were amplified by polymerase chain reaction (PCR). The probes used FISH hybridized to centromere of the X chromosome and heterochromatin region (Yq12) of the Y chromosome, and all PCR related Y chromosome showed positive band like normal male. From the results obtained, it seemed that the Y chromosome from the 46,XY female was structurely normal. Especially, the SRY gene has been equated with the mammalian testis-determining factor, and absence or point mutation in the SRY gene causes XY female. To detect the point mutations of SRY sequences, single-strand conformation polymorphism (SSCP) assay was used. Our results confirm that this patient has no mutation in the SRY gene on the Y chromosome.

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Challenges of Genome Wide Sequencing Technologies in Prenatal Medicine (산전 진단에서의 염기 서열 분석 방법의 의의)

  • Kang, Ji-Un
    • The Journal of the Korea Contents Association
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    • v.22 no.2
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    • pp.762-769
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    • 2022
  • Genetic testing in prenatal diagnosis is a precious tool providing valuable information in clinical management and parental decision-making. For the last year, cytogenetic testing methods, such as G-banding karyotype analysis, fluorescent in situ hybridization, chromosomal microarray, and gene panels have evolved to become part of routine laboratory testing. However, the limitations of each of these methods demonstrate the need for a revolutionary technology that can alleviate the need for multiple technologies. The recent introduction of new genomic technologies based on next-generation sequencing has changed the current practice of prenatal testing. The promise of these innovations lies in the fast and cost-effective generation of genome-scale sequence data with exquisite resolution and accuracy for prenatal diagnosis. Here, we review the current state of sequencing-based pediatric diagnostics, associated challenges, as well as future prospects.

Analysis of Attached Bacterial Community of Biological Activated Carbon Process Using FISH (FISH 기법을 이용한 생물활성탄 공정에서의 운전기간별 부착 박테리아 군집변화 분석)

  • Son, Hyeng-Sik;Son, Hee-Jong;Park, Geun-Tae;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.22 no.1
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    • pp.25-35
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    • 2013
  • The concentration of organic compounds was analyzed at each step of BAC process though $BDOC_{total/rapid/slow}$. Further, bacteria communities and biomass concentrations measured FISH and ATP methods were analyzed. The bed volume (BV) of steady state is different from that of based on assessment of organic compounds removal. Bed volumes in DOC, $BDOC_{rapid}$ and $BDOC_{total/slow}$ removal at steady state were around 27,500 (185.8 day), 15,000 (101.4 day) and 32,000 (216.2 day), respectively. A biomass didn't change after the bed volume reached 22,500 (152.0 day) according to analyzing ATP concentration of bacteria. The concentration of ATP was 2.14 ${\mu}g/g$ in BV 22,500 (152.0 day). The total bacterial number was $4.01{\pm}0.4{\times}10^7$ cells/g at the bed volume 1,150 (7.8 day) (the initial operation) and the number of bacteria was $9.27{\pm}0.2{\times}10^9$ at the bed volume 58,560 395.7 day) that increased more than 200 times. Bacterial uptrend was reduced and bacterial communities were stabilized since BV 18,720 (126.5 day). When BV were 1,150 (7.8 day), 8,916 (60.2 day), 18,720 (126.5 day), 31,005 (209.5 day), 49,632 (335.3 day), 58,560 (395.7 day), a proportion of total bacteria for the Eubacteria were 60.1%, 66.0%, 78.4%, 82.0%, 81.3% respectively. ${\gamma}$-Proteobacteria group was the most population throughout the entire range. The correlation coefficient ($r^2$) between Eubacteria biomass and ATP concentration was 0.9448.