• Tuberculosis and Respiratory Diseases
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• 제50권1호
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• pp.12-28
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• 2001

배경 : 면역 증강성 CpG 올리고 뉴클레오티드(immunostimulatory CpG-oligodeoxynucleotides, ISS CpG-ODN)는 $T_{H1}$ 형 면역 반응은 항진시키고, $T_{H2}$ 형의 반응은 감소시키는 것으로 보고되고 있다. 본 연구에서는 백서 천식 모델에서 ISS CpG-ODN으로 인한 면역 반응의 변화가 천식의 주요한 병태 생리 현상인, 기도 과민성, 호산구성 염증 및 점액 과다 분비에 미치는 효과에 대하여 고찰하고자 하였다. 대상 및 방법 : 천식 모델을 형성하지 않은 10마리의 정상 대조군과 ovalbumin(OVA)의 감작 및 유발로 천식 모델이 형성된 26마리의 백서를, 천식 대조군 11마리, 천식-CpG 치료군 15마리로 나누어 실험하였다. 천식-CpG 치료군은 연구 16일과 20일에 ISS CpG-ODN을, 천식 대조군은 생리 식염수를 투여하였다. CpG-ODN 혹은 생리 식염수 투여 후, 특이 기도 저항, 기관지 폐포 세척액(bronchoalveolar lavage fluid, BALF) 내의 $T_{H1}$ 형 사이토카인 (cytokine)인 IFN-${\gamma}$와 $T_{H2}$ 형 사이토카인인 IL-4의 농도 및 호산구의 분획, 폐조직 내 호산구의 침윤 및 배상 세포의 이형성과 MUC5AC 유전자의 발현을 고찰하였다. 결과 : ISS CpG-ODN 투여 후 천식 대조군과 비교하여, 천식-CpG 치료군에서 기관지 폐포 세척액 내에서 IFN-${\gamma}$의 농도는 유의하게 높았고, IL-4의 농도는 유의하게 낮았다. 특이 기도 저항, 기관지 폐포 세척액 내의 호산구의 분획 및 폐조직에서의 호산구의 침윤은 천식-CpG 치료군에서 천식 대조군에 비하여 유의하게 감소하였다. 그러나 점액 과다 분비의 지표인 배상 세포의 이형성 및 MUC5AC 유전자의 발현 정도는 천식 대조군과 천식-CpG 치료군간에 유의한 차이가 없었다. 결론 : 백서 천식 모델에서 면역 증강성 CpG 올리고 뉴클레오티드는 $T_{H1}$ 형 면역 반응은 항진시키고, $T_{H2}$ 형의 반응은 감소시킴으로써, 천신의 주요한 병태 생리 현상인 기도 과민성 및 호산구서 염증은 억제하나, 점액과다 분비에는 유의한 영향이 없었다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권11호
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• pp.1532-1539
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• 2014

Although growth rate is one of the main economic traits of concern in pig production, there is limited knowledge on its epigenetic regulation, such as DNA methylation. In this study, we conducted methyl-CpG binding domain protein-enriched genome sequencing (MBD-seq) to compare genome-wide DNA methylation profile of small intestine and liver tissue between fast- and slow-growing weaning piglets. The genome-wide methylation pattern between the two different growing groups showed similar proportion of CpG (regions of DNA where a cytosine nucleotide occurs next to a guanine nucleotide in the linear sequence) coverage, genomic regions, and gene regions. Differentially methylated regions and genes were also identified for downstream analysis. In canonical pathway analysis using differentially methylated genes, pathways (triacylglycerol pathway, some cell cycle related pathways, and insulin receptor signaling pathway) expected to be related to growth rate were enriched in the two organ tissues. Differentially methylated genes were also organized in gene networks related to the cellular development, growth, and carbohydrate metabolism. Even though further study is required, the result of this study may contribute to the understanding of epigenetic regulation in pig growth.

• Molecules and Cells
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• 제39권12호
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• pp.888-897
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• 2016

Stable expression of Foxp3 is ensured by demethylation of CpG motifs in the Foxp3 intronic element, the conserved non-coding sequence 2 (CNS2), which persists throughout the lifespan of regulatory T cells (Tregs). However, little is known about the mechanisms on how CNS2 demethylation is sustained. In this study, we found that Ten-Eleven-Translocation (Tet) DNA dioxygenase protects the CpG motifs of CNS2 from re-methylation by DNA methyltransferases (Dnmts) and prevents Tregs from losing Foxp3 expression under inflammatory conditions. Upon stimulation of Tregs by interleukin-6 (IL6), Dnmt1 was recruited to CNS2 and induced methylation, which was inhibited by Tet2 recruited by IL2. Tet2 prevented CNS2 re-methylation by not only the occupancy of the CNS2 locus but also by its enzymatic activity. These results show that the CNS2 methylation status is dynamically regulated by a balance between Tets and Dnmts which influences the expression of Foxp3 in Tregs.

• IMMUNE NETWORK
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• 제15권1호
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• pp.9-15
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• 2015

Streptococcus pneumoniae (the pneumococcus) causes a major upper respiratory tract infection often leading to severe illness and death in the elderly. Thus, it is important to induce safe and effective mucosal immunity against this pathogen in order to prevent pnuemocaccal infection. However, this is a very difficult task to elicit protective mucosal IgA antibody responses in older individuals. A combind nasal adjuvant consisting of a plasmid encoding the Flt3 ligand cDNA (pFL) and CpG oligonucleotide (CpG ODN) successfully enhanced S. pneumoniae-specific mucosal immunity in aged mice. In particular, a pneumococcal surface protein A-based nasal vaccine given with pFL and CpG ODN induced complete protection from S. pneumoniae infection. These results show that nasal delivery of a combined DNA adjuvant offers an attractive potential for protection against the pneumococcus in the elderly.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.181-184
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• 2010

Osmolarity of culture media is one of the most important factors affecting in vitro development. This study was conducted to investigate the DNA methylation status of Pre-1 and satellite sequence 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 NaG or 0.05M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. Previous our studies have reported that pNT embryos cultured in both hypertonic media showed significantly higher blastocyst formation rate than that of control. The DNA methylation status of the satellite sequences in blastocyst was characterized using bisulfite-sequencing technology. The satellite region had a similar methylation pattern of in vivo blastocyst among two culture groups excepting the control group. Each level of methylation is that the satellite DNA moderately methylated (43.10% of PZM-3; 56.12% of NaCl; 55.06% of sucrose; 60.00% of in vivo embryos). As a result of the sequence of PRE-1, CpG methylation pattern was similar to three groups, including in vivo group. In case of the satellite DNA region, the osmolarity conditions were affected CpG DNA methylation status while PRE-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. These results indicate that the modification of osmolarity in a culture media may influence to spatially change of DNA methylation of repetitive sequence for pNT embryo development.

• Clinical and Experimental Pediatrics
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• 제45권7호
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• pp.875-883
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• 2002

목 적 : 기관지 천식은 Th2 반응 우세로 인한 기도의 호산구증과 IgE 증가를 특징으로 하는 질환이다. 박테리아 DNA 혹은 합성 ODNs인 unmethylated CpG motifs(CpG)은 Th1 사이토카인인 $INF-{\gamma}$와 IL-12 유도와 Th2 사이토카인을 하강 조절하여 기도 내의 호산구증 및 전신적 IgE 생성을 억제하며, 메타콜린 흡입에 의한 기관지 과민성을 방지한다. 저자들은 기도 염증을 유발한 생쥐의 비강내에 합성 CpG ODNs의 점적 투여로 기도의 호산구증과 그 외 면역학적 기전에 대한 영향을 연구하고자 한다. 방 법 : BALB/c 생쥐에 주당 2번씩 면역보강제 없이 OVA를 $10{\mu}g$을 피하 주사하여 감작 시켰다. 10주째에 $50{\mu}g$ OVA를 백서의 비강내로 주입하여 기도 염증을 유도한 후, 4주 뒤인 14주째 다시 생쥐의 비강내에 OVA＋CpG ODNs과 OVA를 점적 흡입 주입하였다. 마지막 항원 유발(14주) 후 4일째에 기관지 폐포 세척액을 추출하여 세포를 염색한 후 호산구를 측정하였고 동시에 비장 세포를 배양하여 사이토카인 측정을 하였다. 비장 세포에서 OVA에 의해 생성된 사이토카인 즉 Th1($INF-{\gamma}$, IL-12), Th2(IL-4, IL-5, IL-13) 사이토카인을 ELISA에 의해 측정하였다. 혈청 항원 특이 IgE, IgG2a 또한 ELISA에 의해 측정하였다. 결 과 : 1) 기관지폐포 세척액의 호중구수 : CpG ODNs을 투여한 군이 7.6[5.8-9.4]%, CpG ODNs을 투여하지 않은 군이 42.0[38.2-44.6]%로 기도 염증 세포내의 호산구증을 현저히 억제시켰다(P<0.01). 2) 혈청 OVA-specific IgE와 IgG2a : CpG ODNs으로 투여한 군과 투여하지 않은 군에서 측정한 혈청 OVA-specific IgE와 IgG2a는 음성 대조군에 비해 현저히 증가하였고, CpG ODNs을 비강내 투여 후 4일째 측정한 혈청 OVA-specific IgE와 IgG2a 수치는 CpG ODNs 투여군과 비투여군 사이에 유의한 차이가 없었다. 3) 사이토카인 분석 : CpG ODNs으로 투여한 군은 투여하지 않은 군에 비해 IL-4, IL-5, IL-13의 생성이 감소하였고 IL-12 생성은 증가하였으나, $INF-{\gamma}$의 상향조절은 보여주지 못하였다. 결 론: CpG ODNs의 백신 접종은 면역반응을 조절하여 기도 염증이 형성된 생쥐에서 기도 호산구증을 감소시키는데 매우 효과적이며 천식 치료에 유용할 수 있을 것으로 사료된다.

• Genomics & Informatics
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• 제15권1호
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• pp.28-37
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• 2017

Obesity is a highly prevalent, chronic disorder that has been increasing in incidence in young patients. Both epigenetic and genetic aberrations may play a role in the pathogenesis of obesity. Therefore, in-depth epigenomic and genomic analyses will advance our understanding of the detailed molecular mechanisms underlying obesity and aid in the selection of potential biomarkers for obesity in youth. Here, we performed microarray-based DNA methylation and gene expression profiling of peripheral white blood cells obtained from six young, obese individuals and six healthy controls. We observed that the hierarchical clustering of DNA methylation, but not gene expression, clearly segregates the obese individuals from the controls, suggesting that the metabolic disturbance that occurs as a result of obesity at a young age may affect the DNA methylation of peripheral blood cells without accompanying transcriptional changes. To examine the genome-wide differences in the DNA methylation profiles of young obese and control individuals, we identified differentially methylated CpG sites and investigated their genomic and epigenomic contexts. The aberrant DNA methylation patterns in obese individuals can be summarized as relative gains and losses of DNA methylation in gene promoters and gene bodies, respectively. We also observed that the CpG islands of obese individuals are more susceptible to DNA methylation compared to controls. Our pilot study suggests that the genome-wide aberrant DNA methylation patterns of obese individuals may advance not only our understanding of the epigenomic pathogenesis but also early screening of obesity in youth.

• 식물분류학회지
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• 제46권2호
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• pp.163-174
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• 2016

Gnetum (Gnetaceae, Gnetales) is a gymnosperm genus with ca. 35 species distributed in tropical forests around the world. Due to its dioecious habit and lack of diagnostic characters from vegetative tissue, the identification of Gnetum species is not easy without seeds or reproductive structures. To identify and verify their phylogenetic positions, we applied DNA barcoding to Cambodian Gnetum collections gathered between 2010 and 2015, with previously designed cp matK gene primers. We newly sequenced partial matK sequences from 72 Gnetum collections, 43 out of 72 from Cambodia, and analyzed 115 Gnetum accessions using the neighbor-joining method. The resulting neighbor-joining tree categorized Cambodian Gnetum samples into three clades of species: G. macrostachyum, G. montanum, and G. aff. gracilipes. The recognition of G. aff. gracilipes in Cambodia is reported here for the first time. Taxonomic information for the three recognized Cambodian Gnetum species is provided and the benefits of the taxonomic reevaluation assisted by DNA barcoding are emphasized in this work.

• Genomics & Informatics
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• 제9권3호
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• pp.136-137
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• 2011

Methylation of cytosine is a post-synthesis modification that does not affect the primary DNA sequence but greatly influences gene expression level and phenotypes of an organism. As high-throughput sequencing of bisulfite-treated DNA is the most efficient method to identify methylated sites, several tools to map sequencing reads on a reference are available. But tools to visualize and to interpret the methylation level of methylation sites are currently insufficient. Herein, we present a novel tool to visualize the methylation level of CpG sites.

• International Journal of Industrial Entomology and Biomaterials
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• 제8권1호
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• pp.95-99
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• 2004

Synthetic oligodeoxynucleotides (ODNs) containing unmethylated CpG dinucleotides in particular base contexts are known to induce immunity in vertebrate cells. In insect, however, it was recent to find out that ODNs induces insect immunity as other immune inducer such as lipopolysaccharide. However, the finding was solely based on one lepidopteran insect, Bombyx mori, and the expression of insect immunity was neither dependent on numbers of CpG repeats nor methylation of CpG repeats within ODNs. Instead, foreignness of DNA has been suggested to be a key factor governing induction of antibacterial peptide. In this study, we expanded our previous understanding to the potentiality of ODNs as an immune inducer for antifungal peptide in Galleria mellonella and B. mori. To do this, a defensin-type antifungal peptide gene, reported from G. mellonella was cloned and partially sequenced from G. mellonella and B. mori successfully and utilized as a probe in the Northern blot analysis. We found out that ODNs also work as an immune inducer for antifungal peptide in the fat body and midgut of G. mellonella and B. mori larvae. Also, induction pattern of antifungal peptide was irrelevant to the numbers of CpG repeats within ODNs as previously reported on the induction pattern of antibacterial peptides.