• 제목/요약/키워드: DNA data

검색결과 2,054건 처리시간 0.028초

Immune Responses Induced by HSP60 DNA Vaccine against Toxoplasma gondii Infection in Kunming Mice

  • Li, Zhong-Yuan;Lu, Jing;Zhang, Nian-Zhang;Chen, Jia;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • 제56권3호
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    • pp.237-245
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    • 2018
  • Toxoplasma gondii can infect all the vertebrates including human, and leads to serious toxoplasmosis and considerable veterinary problems. T. gondii heat shock protein 60 (HSP60) is associated with the activation of antigen presenting cells by inducing initial immune responses and releasing inflammatory cytokines. It might be a potential DNA vaccine candidate for this parasite. A pVAX-HSP60 DNA vaccine was constructed and immune responses was evaluated in Kunming mice in this study. Our data indicated that the innate and adaptive immune responses was elicited by successive immunizations with pVAX-HSP60 DNA, showing apparent increases of CD3e+CD4+ and CD3e+CD8a+ T cells in spleen tissues of the HSP60 DNA-immunized mice ($24.70{\pm}1.23%$ and $10.90{\pm}0.89%$, P<0.05) and higher levels of specific antibodies in sera. Furthermore, the survival period of the immunized mice ($10.53{\pm}4.78day$) were significantly prolonged during the acute T. gondii infection. Decrease of brain cysts was significant in the experimental group during the chronic infection (P<0.01). Taken together, TgHSP60 DNA can be as a vaccine candidate to prevent the acute and chronic T. gondii infections.

아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용 (Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • 제35권10호
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

Novel non-invasive molecular identification method for two tree frogs, Dryophytes suweonensis and Dryophytes japonicus, based on high resolution melting(HRM) analysis

  • Nakyung Yoo;Keun-Yong Kim;Jung Soo Heo;Ju-Duk Yoon;Keun-Sik Kim
    • 환경생물
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    • 제40권2호
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    • pp.199-205
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    • 2022
  • Two tree frogs, Dryophytes suweonensis and Dryophytes japonicus, inhabiting Korea, are morphologically similar and share the same habitats. Therefore, they are identified mainly through their calls, especially for males. Dryophytes suweonensis is registered as an endangered (IUCN: EN grade) and protected species in South Korea. Thus, it is necessary to develop a method to rapidly identify and discriminate the two species and establish efficient protection and restoration plans. We identified significant genetic variation between them by sequencing a maternally-inherited mitochondrial 12S ribosomal DNA region. Based on the sequence data, we designed a pair of primers containing 7bp differences for high resolution melting(HRM) analysis to rapidly and accurately characterize their genotypes. The HRM analysis using genomic DNA showed that the melting peak for D. suweonensis was 76.4±0.06℃, whereas that of D. japonicus was 75.0±0.05℃. The differential melt curve plot further showed a distinct difference between them. We also carried out a pilot test for the application of HRM analysis based on immersing D. suweonensis in distilled water for 30 min to generate artificial environmental DNA(eDNA). The results showed 1.10-1.31℃ differences in the melting peaks between the two tree frog samples. Therefore, this HRM analysis is rapid and accurate in identifying two tree frogs not only using their genomic DNA but also using highly non-invasive eDNA.

Effects of Pogonatherum paniceum (Lamk) Hack extract on anti-mitochondrial DNA mediated inflammation by attenuating Tlr9 expression in LPS-induced macrophages

  • Rungthip Thongboontho;Kanoktip Petcharat;Narongsuk Munkong;Chakkraphong Khonthun;Atirada Boondech;Kanokkarn Phromnoi;Arthid Thim-uam
    • Nutrition Research and Practice
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    • 제17권5호
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    • pp.827-843
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    • 2023
  • BACKGROUND/OBJECTIVES: Mitochondrial DNA leakage leads to inflammatory responses via endosome activation. This study aims to evaluate whether the perennial grass water extract (Pogonatherum panicum) ameliorate mitochondrial DNA (mtDNA) leakage. MATERIALS/METHODS: The major bioactive constituents of P. paniceum (PPW) were investigated by high-performance liquid chromatography, after which their antioxidant activities were assessed. In addition, RAW 264.7 macrophages were stimulated with lipopolysaccharide, resulting in mitochondrial damage. Quantitative polymerase chain reaction and enzyme-linked immunosorbent assay were used to examine the gene expression and cytokines. RESULTS: Our results showed that PPW extract-treated activated cells significantly decrease reactive oxygen species and nitric oxide levels by reducing the p2phox and iNOS expression and lowering cytokine-encoding genes, including IL-6, TNF-α, IL-1β, PG-E2 and IFN-γ relative to the lipopolysaccharide (LPS)-activated macrophages. Furthermore, we observed that LPS enhanced the mtDNA leaked into the cytoplasm, increasing the transcription of Tlr9 and signaling both MyD88/Irf7-dependent interferon and MyD88/NF-κb p65-dependent inflammatory cytokine mRNA expression but which was alleviated in the presence of PPW extract. CONCLUSIONS: Our data show that PPW extract has antioxidant and anti-inflammatory activities by facilitating mtDNA leakage and lowering the Tlr9 expression and signaling activation.

그룹 구조를 갖는 고차원 유전체 자료 분석을 위한 네트워크 기반의 규제화 방법 (Network-based regularization for analysis of high-dimensional genomic data with group structure)

  • 김기풍;최지윤;선호근
    • 응용통계연구
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    • 제29권6호
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    • pp.1117-1128
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    • 2016
  • 고차원 유전체 자료를 사용하는 유전체 연관 분석에서는 벌점 우도함수 기반의 회귀계수 규제화 방법이 질병 및 표현형질에 영향을 주는 유전자를 발견하는데 많이 이용된다. 특히, 네트워크 기반의 규제화 방법은 유전체 연관성 연구에서의 유전체 경로나 신호 전달 경로와 같은 생물학적 네트워크 정보를 사용할 수 있으므로, Lasso나 Elastic-net과 같은 다른 규제화 방법들과 비교했을 경우 네트워크 기반의 규제화 방법이 보다 더 정확하게 관련 유전자들을 찾아낼 수 있다는 장점을 가지고 있다. 그러나 네트워크 기반의 규제화 방법은 그룹 구조를 갖고 있는 고차원 유전체 자료에는 적용시킬 수 없다는 문제점을 가지고 있다. 실제 SNP 데이터와 DNA 메틸화 데이터처럼 대다수의 고차원 유전체 자료는 그룹 구조를 가지고 있으므로 본 논문에서는 이러한 그룹 구조를 가지고 있는 고차원 유전체 자료를 분석하고자 네트워크 기반의 규제화 방법에 주성분 분석(principal component analysis; PCA)과 부분 최소 자승법(partial least square; PLS)과 같은 차원 축소 방법을 결합시키는 새로운 분석 방법을 제안하고자 한다. 새롭게 제안한 분석 방법은 몇 가지의 모의실험을 통해 변수 선택의 우수성을 입증하였으며, 또한 152명의 정상인들과 123명의 난소암 환자들로 구성된 고차원 DNA 메틸화 자료 분석에도 사용하였다. DNA 메틸화 자료는 대략 20,000여개의 CpG sites가 12,770개의 유전자에 포함되어 있는 그룹 구조를 가지고 있으며 Illumina Innium uman Methylation27 BeadChip으로부터 생성되었다. 분석 결과 우리는 실제로 암에 연관된 몇 가지의 유전자를 발견할 수 있었다.