• Title/Summary/Keyword: Nuclear Sites

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Methylation Pattern of H19 Gene at Various Preimplantation Development Stages of In Vitro Fertilized and Cloned Porcine Embryos

  • Im, Young-Bin;Han, Dong-Wook;Gupta, Mukesh Kumar;Uhm, Sang-Jun;Heo, Young-Tae;Kim, Jin-Hoi;Park, Chan-Kyu;Lee, Hoon-Taek
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.83-90
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    • 2007
  • Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, $8{\sim}16$-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and $8{\sim}16$-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted ($0{\sim}2%$) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-,4- and $8{\sim}16$-cell and blastocyst stages of development in both IVF ($0{\sim}14.1%$) and SCNT ($0{\sim}6.4%$) embryos. At all stages of development, CTCF3 was unmethylated in IVF ($0{\sim}17.3%$) and SCNT ($0{\sim}1.2%$) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.

Non-CpG Methylation of Pre-1 Sequence in Pig SCNT Blastocysts (돼지 체세포복제 배반포에서 Pre-1 영역의 Non-CpG 메틸화 양상)

  • Ko, Yeoung-Gyu;Im, Gi-Sun;Lee, Hwi-Cheul;Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Lee, Poong-Yeon;Cho, Chang-Yeon;Cho, Jae-Hyeon;Yoo, Young-Hee
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.93-97
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    • 2011
  • Previously, we reported that the osmolarity conditions in the satellite region were affected CpG DNA methylation status while Pre-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. This study was conducted to investigate the DNA methylation status of repeat sequences in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaCl or 0.05 M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. The DNA methylation status of the Pre-1 sequences in blastocysts was characterized using a bisulfite-sequencing method. Intriguingly, in the present study, we found the unique DNA methylation at several non-CpG sequences at the Pre-1 sequences in all groups. The non-CpG methylation was hypermethylated in all three groups, including in vivo group (86.90% of PZM-3; 83.87% of NaCl; 84.82% of sucrose; 90.94% of in vivo embryos). To determine whether certain non-CpG methylated sites were preferentially methylated, we also investigated the methylation degree of CpA, CpT and CpC. Excepting in vivo group, preference of methylation was CpT>CpC>CpA in all three groups investigated. These results indicate that DNA methylation of Pre-1 sequences was hypermethylated in CpG as well as non-CpG site, regardless modification of osmolarity in a culture media.

Metabolic impairment pattern analysis of the Alzheimer's disease (Alzheimer's Disease의 대사영상패턴 분석)

  • Juh, Ra-Hyeong;Lee, Chang-Uk;Chung, Yong-An;Choe, Bo-Young;Suh, Tae-Suk
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2004.11a
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    • pp.91-95
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    • 2004
  • Several MRI studies have reported reductions in temporal lobe volumes in Alzheimer's disease (AD). Measures have been usually obtained with regions-of-interest (ROI) drawn manually on selected medial and lateral portions of the temporal lobes, with variable choices of anatomical borders across different studies. We used the automated voxel-based morphometry (VBM) approach to investigate gray matter abnormalities over the entire extension of the temporal lobe in 10AD patients (MM5E 22)and 22 healthy controls. Foci of significantly reduced gray matter volume in AD patients were detected in both medial and lateral temporal regions, most significantly in the right and left posterior parahippocarmpal gyri. At a more flexible statistical threshold (P<0.01, uncorrected for multiple comparisons), circumscribed foci of significant gray matter reduction were also detected in the right amygdala/enthorinal cortex, the anterior and posterior borders of the superior temporal gyrus bilaterally, and the anterior portion of the left middle temporal gyrus. These VBM results confirm previous findings of temporal lobe atrophic changes in AD, and suggest that these abnormalities may be confined to specific sites within that lobe, rather than showing a widespread distribution.

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Translocation of Protein Kinase C Isozymes in the Breast Cancer Cell Line (유방 암세포에서 Protein Kinase C 동위효소의 전위)

  • Won Chul Choi;Joo Young Son;Seok Jin Seo
    • Journal of Life Science
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    • v.8 no.6
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    • pp.638-647
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    • 1998
  • Protein Kinase C (PKC) activators, phorbol 12-myristate 13-acetate (PMA), bryostatin, and dioctanoyl glycerol (DiC8), induce translocation of PKC isozymes from cytoplasm to plasma membrane or into nucleus. The activated PKC negatively modulates growth of human breast cancer cells. Antiproliferative effect and translocation of PKC were investigated in MCF-7 cells. The translocation of activated PKC isozymes by PMA, bryostatin and DiC8 was occurred at the various different regions in MCF-7 cell. PKC $\alpha$ and $\beta$ could be translocated to the nucleus or the nuclear mem-brane, and PKC $\delta$and $\varepsilon$ to cell membrane by PMA while DiC8 and bryostatin induced the translocation of PKC $\alpha$ and $\beta$ to the nucleus or plasma membrane, respectively. In the antiproliferative effect of PKC activators, PMA ($IC_{50}$/ values of 1.2$\pm$0.3nM) and DiC8 ($IC_{50}$/ values of 5.0$\pm$1.1$\mu$M) inhibited the cell growth. Bryostatin also inhibited the cell growth but to a much less degree than one obser-ved with PMA : 16% growth reduction by 100nM bryostatin. However, PMA treated with bryostatin induced gro-wth inhibition, but PMA with DiC8 at 10$\mu$M was not effective. These results suggest that each PKC isozyme is tran-slocated to various specific sites, and that especially, PKC $\alpha$ isozyme plays an important role in control of antiprolife-raive function of cell growth.

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A Study on Continuous long-term Wave Observation using Remote Monitoring System (원격모니터링을 이용한 연속파랑관측에 관한 연구)

  • Shin, Bumshick
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.1
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    • pp.654-659
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    • 2018
  • In this study, continuous long-term observation is implemented with an ocean radar. Ocean radar conducts remote observation (combined) with ground-based radars, which enable a series of simultaneous observations of an extensive range of the coast with high frequency. An ocean radar for continuous long-term observation is operated at Samcheok on the east coast of Korea. Samcheok experienced tsunami damage in recent years and is the location of a nuclear power plant. In order to examine the reliability of the ocean radar, a pressure-type wave gauge, ultrasonic wave gauge, and ocean buoy are installed for the purpose of data comparison and verification. The ocean radar used in this study is an array-type HF-RADAR named WERA (WavE RAdar). The analysis of the data obtained from continuous long-term observations showed that the radar observations were in agreement with more than 90% of the wave data collected within a 25 km range from the center of two sites. Less than 1% of the entire observation data was unmeasured by the time series analysis. As a result of comparing the radar data with the direct observations made by the wave gauge, it was inferred that the RMS deviation is less than 20cm and the correlation coefficient was in the range of 0.84 ~ 0.87. Moreover, supported by such observations, a comprehensive monitoring system is being developed to provide the public with real-time reports on waves and currents via the internet.

Soil-to-Rice Seeds Transfer Factors of Radioiodine and Technetium for Paddy Fields around the Radioactive-Waste Disposal Site in Gyeongju (경주 방사성 폐기물 처분장 주변 논에 대한 방사성 요오드와 테크네튬의 토양-쌀알 전이계수)

  • Choi, Yong-Ho;Lim, Kwang-Muk;Jun, In;Park, Doo-Won;Keum, Dong-Kwon;Han, Moon-Hee
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.8 no.4
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    • pp.329-337
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    • 2010
  • Radiotracer experiments were performed over two years using pot cultures in a greenhouse to investigate soil-torice seeds transfer factors of radioiodine and technetium for paddy fields around the radioactive-waste disposal site in Gyeongju. Before transplanting rice seedlings, the top about 20 cm soils were thoroughly mixed with $^{125}I$ (2007) and $^{99}Tc$ (2008), and the pots were irrigated to simulate flooded rice fields. Transfer factors were determined as the ratios of the radionuclide concentrations in dry rice seeds (brown rice) to those in dry soils. Transfer factors of radioiodine and technetium were in the ranges of $1.1{\times}10^{-3}{\sim}6.4{\times}10^{-3}$ (three soils) and $5.4{\times}10^{-4}{\sim}2.5{\times}10^{-3}$ (four soils), respectively, for different soils. It seems that the differences in the clay content among soils played a more important role for such variations than those in the organic matter content and pH. As the representative values of radioiodine and technetium transfer factors for rice seeds, $2.9{\times}10^{-3}$ and $1.1{\times}10^{-3}$, respectively, were proposed. In order to obtain more highly representative values in the future, investigations for the sites of interest need to be carried out continuously.

Natural hybridization of Iris species in Mt. Palgong-san, Korea (팔공산 금붓꽃 계열의 자연 잡종 현상)

  • Son, OGyeong;Son, Sung-Won;Suh, Gang-Uk;Park, SeonJoo
    • Korean Journal of Plant Taxonomy
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    • v.45 no.3
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    • pp.243-253
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    • 2015
  • Series Chinensis, Genus Iris, endemic to the far regions of East Asia, consists of four species and related varieties. This series is divided into two major groups (I. rossii and I. minutiaurea complex). In this study, the ITS region and matK gene sequences within nuclear ribosomal DNA and plastid DNA were analyzed in order to investigate the phylogenetic relationships among the I. minutiaurea complex (I. minutiaurea, I. odaesanensis, and I. koreana) and the taxonomic identities of a putative hybrid in Mt. Palgong. In the internal transcribed spacer (ITS1, 5.8S, and ITS2) region, a total of 106 cloned genomic sequences from three taxa were obtained to study the intragenomic polymorphisms of the ITS regions. Three taxa revealed high levels of intragenomic polymorphisms, indicative of incomplete nrDNA concerted evolution. This incomplete ITS concerted evolution in the series Chinensis may be linked to the recent species divergence and frequent interspecies hybridization of the series Chinensis. In the matK gene, three taxa were fairly separated by eleven variable sites. In eight individuals collected on Mt. Palgong, putative hybrids between I. odaesanensis and I. minutiaurea were clustered in the I. minutiaurea clade in the NJ (neighbor-joining) tree based on the matK gene. However, in the ITS tree, some of them were clustered in the I. odaesanensis clade and others were clustered in the I. minutiaurea clade. Therefore, the individuals on Mt. Palgong were formed by the hybridization between two taxa (I. odaesanensis and I. minutiaurea) and not through the lineage of I. koreana.

Screening for the 3' UTR Polymorphism of the PXR Gene in South Indian Breast Cancer Patients and its Potential role in Pharmacogenomics

  • Revathidevi, Sundaramoorthy;Sudesh, Ravi;Vaishnavi, Varadharajan;Kaliyanasundaram, Muthukrishnan;MaryHelen, Kilyara George;Sukanya, Ganesan;Munirajan, Arasambattu Kannan
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.8
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    • pp.3971-3977
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    • 2016
  • Background: Breast cancer, the commonest cancer among women in the world, ranks top in India with an incidence rate of 1,45,000 new cases and mortality rate of 70,000 women every year. Chemotherapy outcome for breast cancer is hampered due to poor response and irreversible dose-dependent cardiotoxicity which is determined by genetic variations in drug metabolizing enzymes and transporters. Pregnane X receptor (PXR), a member of the nuclear receptor superfamily, induces expression of drug metabolizing enzymes (DMEs) and transporters leading to regulation of xenobiotic metabolism. Materials and Methods: A genomic region spanning PXR 3' UTR was amplified and sequenced using genomic DNA isolated from 96 South Indian breast cancer patients. Genetic variants observed in our study subjects were queried in miRSNP to establish SNPs that alter miRNA binding sites in PXR 3' UTR. In addition, enrichment analysis was carried out to understand the network of miRNAs and PXR in drug metabolism using DIANA miRpath and miRwalk pathway prediction tools. Results: In this study, we identified SNPs rs3732359, rs3732360, rs1054190, rs1054191 and rs6438550 in the PXR 3; UTR region. The SNPs rs3732360, rs1054190 and rs1054191 were located in the binding site of miR-500a-3p, miR-532-3p and miR-374a-3p resulting in the altered PXR level due to the deregulation of post-transcriptional control and this leads to poor treatment response and toxicity. Conclusions: Genetic variants identified in PXR 3' UTR and their effects on PXR levels through post-transcriptional regulation provide a genetic basis for interindividual variability in treatment response and toxicity associated with chemotherapy.

Two-dimensional Inversion of Sea-effect-corrected Magnetotelluric (MT) Data in Jeju Island (해양효과가 보정된 제주도 자기지전류 탐사 자료의 2차원 역산)

  • Yang, Jun-Mo;Lee, Heui-Soon;Lee, Choon-Ki;Park, Gye-Soon
    • Journal of the Korean earth science society
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    • v.32 no.6
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    • pp.602-612
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    • 2011
  • Jeju Island, a volcanic island located in South Korea, has been one of the main targets of geophysical and/or geological studies because of its tectonic importance related to the volcanism and tectonic link to the southern part of the Korean Peninsula. Recently, as a number of broad-band magnetotelluric (MT) measurements were made, we have examined the deep part of the island. In such an insular setting, it is not easy to properly recover the deep structures such as the lower crust and the upper crust using MT data, because their low-frequency components are strongly affected by the surrounding sea of the island. In this study, we apply the sea-effect correction to the existing MT data collected at a total of 102 sites in Jeju Island. The sea-effect correction makes remarkable changes in the observed MT data at frequencies below 1 Hz, clearly indicating the existence of a conductive lower crust. The 2-D inversion results for both Jeju Southern Line (JSL) and Jeju Northern Line (JNL) show that the transition zone separating the resistive upper crust and conductive lower crust exists at a depth of 20 km on average.

Biodistribution of [S-35] Labeled Antisense Oligodeoxynucleotides Increased Tumor Targeting With Microsphere Coinjection

  • Choe, Jae-Gol;Park, Gil-Hong;Claudio Nastruzzi;Yoon S. Cho-Chung;Kim, Meyoung-Kon
    • Environmental Mutagens and Carcinogens
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    • v.22 no.2
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    • pp.65-69
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    • 2002
  • To elucidate the effect of microsphere coinjection on the administration of oligodeoxynucleotides (ODN), we have investigated biodistribution of [S-35]-labeled antisense ODN targeted to cAMP-dependent protein kinase (PKA) RI-$\alpha$ subunit in nude mice xenografted with WiDr (human colon cancer, ATCC CCL218). The strategy of using microsphere has been proposed for cancer treatment as a carrier of therapeutic ODN so that it could offer an advantage with respect to maintaining constant ODN levels in blood and obtaining higher therapeutic ODN concentration at tumor sites. Comparative biodistribution studies were performed in nude mice (female, 20 g of body weight, n = 4-6) xenografted with WiDr cancer cells, when 0.1 $\mu$Ci (specific activity, 2.94 mCi/$\mu$mole) of [S-35]-labeled RI-$\alpha$ antisense ODN was injected alone or with microsphere (PLG-18, polylactic copolymer with cationic surfactant DDAB18). Peak tumor uptake of [S-35]-labeled ODN was significantly increased from 17.7% (at 6 h) of injected dose per gram of tissue (ID/g) to 42.5% (at 24 h) ID/g when microsphere was coinjected with ODN. The different biodistribution in the kidney accumulation (e.g., 100.2% ID/g for ODN alone and 54.9%/ID/g for microshpere coinjection) may contribute to higher blood concentration (e.g., 21.5%ID/$m\ell$ for ODN alone and 37.5%ID/$m\ell$ for microsphere coinjection) of radiolabeled ODN. Of importance is the fact that the whole body retention of radioactivity increased with microsphere coinjection from 50.8%ID/g to 68.0%ID/g after 24-h of injection. This decreased kidney accumulation and increased whole body retention of [S-35]-labeled ODN resulted in a significant improvement of ODN targeting to the tumor site. In conclusion, the coinjection of microsphere appears to be an important carrier system in vehiculation of antisense oligonucleotide to the tumor tissue in vivo.

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