• Title/Summary/Keyword: Bioinformatics study

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Identification and analysis of microRNAs in Candida albicans (Candida albicans의 마이크로RNA 동정과 분석)

  • Cho, Jin-Hyun;Lee, Heon-Jin
    • Journal of Life Science
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    • v.27 no.12
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    • pp.1494-1499
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    • 2017
  • Oral infection due to Candida albicans is a widely recognized and frequent cause of superficial infections of the oral mucosa (oral candidiasis). Although oral candidiasis is not a life-threatening fungemia, it can cause severe problems in individuals under certain conditions. MicroRNAs (miRNAs) are noncoding, small RNA molecules, which regulate the expression of other genes by inhibiting the translation of target mRNAs. The present study was designed to identify miRNAs in C. albicans and determine their possible roles in this organism. miRNA-sized small RNAs (msRNAs) were cloned in C. albicans by deep sequencing, and their secondary structures were analyzed. All the cloned msRNAs satisfied conditions required to qualify them as miRNAs. Bioinformatics analysis revealed that two of the most highly expressed C. albicans msRNAs, Ca-363 and Ca-2019, were located in the 3' untranslated region of the corticosteroid-binding protein 1 (CBP1) gene in a reverse orientation. miRNA mimics were transformed into C. albicans to investigate their RNA-inhibitory functions. RNA oligonucleotide-transformed C. albicans was then observed by fluorescent microscopy. Quantitative PCR analysis showed that these msRNAs did not inhibit CBP1 gene expression 4 hr and 8 hr after ectopic miRNA transformation. These results suggest that msRNAs in C. albicans possess an miRNA-triggered RNA interference gene-silencing function, which is distinct from that exhibited by other eukaryotic systems.

MicroRNA-3200-5p Promotes Osteosarcoma Cell Invasion via Suppression of BRMS1

  • Li, Gen;Li, Li;Sun, Qi;Wu, Jiezhou;Ge, Wei;Lu, Guanghua;Cai, Ming
    • Molecules and Cells
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    • v.41 no.6
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    • pp.523-531
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    • 2018
  • Tumour metastasis is one of the most serious challenges of cancer as it is the major cause of mortality in patients with solid tumours, including osteosarcoma (OS). In this regard, anti-metastatic genes have potential for metastasis inhibition strategies. Recent evidence showed the importance of breast cancer metastasis suppressor 1 (BRMS1) in control of OS invasiveness, but the regulation of BRMS1 in OS remains largely unknown. Here, we used bioinformatics analyses to predict BRMS1-targeting microRNAs (miRNAs), and the functional binding of miRNAs to BRMS1 mRNA was evaluated using a dual luciferase reporter assay. Among all BRMS1-targeting miRNAs, only miR-151b, miR-7-5p and miR-3200-5p showed significant expression in OS specimens. Specifically, we found that only miR-3200-5p significantly inhibited protein translation of BRMS1 via pairing to the 3'-UTR of the BRMS1 mRNA. Moreover, we detected significantly lower BRMS1 and significantly higher miR-3200-5p in the OS specimens compared to the paired adjacent non-tumour bone tissues. Furthermore, BRMS1 and miR-3200-5p levels were inversely correlated to each other. Low BRMS1 was correlated with metastasis and poor patient survival. In vitro, overexpression of miR-3200-5p significantly decreased BRMS1 levels and promoted OS cell invasion and migration, while depletion of miR-3200-5p significantly increased BRMS1 levels and inhibited OS cell invasion and migration. Thus, our study revealed that miR-3200-5p may be a critical regulator of OS cell invasiveness.

Antifungal Activity of Rheum undulatum on Candida albicans by the Changes in Membrane Permeability (막투과성 변화로 인한 대황의 Candida albicans에 대한 항진균 활성)

  • Lee, Heung-Shick;Kim, Younhee
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.360-367
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    • 2014
  • Candida albicans is an opportunistic and the most prevalent fungal pathogen that can cause superficial and systemic infections in immunocompromised patients. C. albicans can promote the transition from budding yeast to filamentous form, generating biofilms. Infections associated with C. albicans biofilms are frequently resistant to conventional antifungal therapy. Therefore, the development of more effective antifungal drugs related with biofilm formation is required urgently. The roots of Rheum undulatum have been used for medicinal purposes in Korea and China traditionally. The aim of present study was to evaluate the effect of R. undulatum extract upon preformed biofilms of 12 clinical C. albicans isolates and the antifungal activities. Its effect on preformed biofilms was evaluated using XTT reduction assay, and metabolic activity of all tested strains was reduced significantly ($49.4{\pm}6.0%$) at 0.098 mg/ml R. undulatum. The R. undulatum extract blocked the adhesion of C. albicans biofilms to polystyrene surfaces, and damaged the cell membrane integrity of C. albicans which was analyzed by CFDA, AM, and propidium iodide double staining. It caused cell lysis which was observed by Confocal laser scanning and phase contrast microscope after propidium iodide and neutral red staining, respectively. Membrane permeability was changed as evidenced by crystal violet uptake. The data suggest that R. undulatum inhibits biofilm formation by C. albicans, which can be associated with the damage of the cell membrane integrity, the changes in the membrane permeability and the cell lysis of C. albicans.

The Concentrations and TEQ Levels of PCDFs and PCDDs in Human Adipose Tissue and First Breast Milk of Korean (체지방 조직 및 초유중에 잔류되어 있는 다이옥신 함유도에 관한 연구)

  • Kim, Yunje;Lee, Sun Young;Han, Soon Young;Park, Kui Lea;Kil, Kwang Sup;Yang, Kyu Hwan;Kim, Eun Kyung;Kim, Myungsoo
    • Analytical Science and Technology
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    • v.14 no.6
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    • pp.504-509
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    • 2001
  • The concentrations and TEQ levels of PCDFs/PCDDs in human aipose tissue and first breast milk were analyzed by high resolution gas chromatography/high resolution mass spectrometry. The human adipose tissue samples measured in this study have been collected at hospital in Seoul. The total concentration and TEQ level of PCDFs were showed 5.812 pg/g and 1.485 pgTEQ/g. The total concentration and TEQ level of PCDDs were showed 26.648 pg/g and 1.176 pgTEQ/g, respectively. This paper also reported dioxin levels in 20 breast milks of Korean mothers from hospital in Seoul National University. Total concentration and TEQ levels of PCDFs were showed 7.019 pg/mL and 0.177 pgTEQ/mL, respectively. Total concentrations and TEQ levels of PCDDs were showed 14.224 pg/mL and 0.693 pgTEQ/mL, respectively. According to the contribution of dioxin congeners in samples, PCDDs was higher than PCDFs. And OCDD had the highest concentration.

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Investigation of Single Nucleotide Polymorphisms in Porcine Candidate Gene for Growth and Meat Quality Traits in the Berkshire Breed (버크셔 품종의 돼지 성장과 육질관련 후보유전자의 단일염기 다형성에 관한 연구)

  • Kim, Sang-Wook;Jung, Ji-Hye;Do, Kyung-Tag;Kim, Kwan-Suk;Do, Chang-Hee;Park, Jun-Kyu;Joo, Young-Kuk;Kim, Tae-Suk;Choi, Bong-Hwan;Kim, Tae-Hun;Song, Ki-Duk;Cho, Byung-Wook
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1622-1626
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    • 2007
  • This study was conducted to identify useful single nucleotide polymorphisms (SNPs) and determine their association with economically important traits in pig population. Four candidate gene analyses have identified important chromosomal regions and major genes associcated whit economic traits of the pig. For application of the chromosomal information to the pig industry using DNA technology, SNP markers were developed by comparative re-sequencing of polymerase chain reaction (PCR) products of 4 candidate genes (CSF2, IL4, MYOD, RIP140). PCR restriction fragment length polymorphism (PCR-RFLP) assays were developed for these 4 SNPs and used to genotype Berkshire pig populations in Korea.

Development of HLA-A, -B and -DR Typing Method Using Next-Generation Sequencing (차세대염기서열분석법을 이용한 HLA-A, -B 그리고 -DR 형별 분석법 개발)

  • Seo, Dong Hee;Lee, Jeong Min;Park, Mi Ok;Lee, Hyun Ju;Moon, Seo Yoon;Oh, Mijin;Kim, So Young;Lee, Sang-Heon;Hyeong, Ki-Eun;Hu, Hae-Jin;Cho, Dae-Yeon
    • The Korean Journal of Blood Transfusion
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    • v.29 no.3
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    • pp.310-319
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    • 2018
  • Background: Research on next-generation sequencing (NGS)-based HLA typing is active. To resolve the phase ambiguity and long turn-around-time of conventional high resolution HLA typing, this study developed a NGS-based high resolution HLA typing method that can handle large-scale samples within an efficient testing time. Methods: For HLA NGS, the condition of nucleic acid extraction, library construction, PCR mechanism, and HLA typing with bioinformatics were developed. To confirm the accuracy of the NGS-based HLA typing method, the results of 192 samples HLA typed by SSOP and 28 samples typed by SBT compared to NGS-based HLA-A, -B and -DR typing. Results: DNA library construction through two-step PCR, NGS sequencing with MiSeq (Illumina Inc., San Diego, USA), and the data analysis platform were established. NGS-based HLA typing results were compatible with known HLA types from 220 blood samples. Conclusion: The NSG-based HLA typing method could handle large volume samples with high-throughput. Therefore, it would be useful for HLA typing of bone marrow donation volunteers.

Growth of Candida albicans Biofilm is Inhibited by Salvia miltiorrhiza (단삼에 의한 Candida albicans 바이오필름 발달의 억제)

  • Lee, Heung-Shick;Kim, Younhee
    • Microbiology and Biotechnology Letters
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    • v.47 no.3
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    • pp.465-472
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    • 2019
  • Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

Molecular Characterization and Expression Analysis of Nucleoporin 210 (Nup210) in Chicken

  • Ndimukaga, Marc;Bigirwa, Godfrey;Lee, Seokhyun;Lee, Raham;Oh, Jae-Don
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.185-191
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    • 2019
  • Nucleoporin 210 (Nup210) is associated with several physiological processes including muscle and neural cell differentiation, autoimmune diseases, and peripheral T cell homeostasis. Chicken Nup210 (chNup210) gene was originally identified as one of the differentially expressed genes (DEGs) in the kidney tissues of chicken. To elucidate the role of Nup210 in metabolic disease of chicken, we studied the molecular characteristics of chNup210 and analyzed its gene expression under the stimulation of Toll-like receptor 3 (TLR3) ligands. The Nup210 genomic DNA and amino acid sequences of various species including fowls, fishes, and mammals were retrieved from the Ensemble database and subjected to bioinformatics analyses. The expression of Nup210 from several chicken tissues was probed through qRT-PCR, and chicken fibroblast DF-1 cell line was used to determine the change in expression of chNup210 after stimulation with TLR3 ligand, polyinosinic-polycytidylic acid (poly (I:C)). The chNup210 gene was highly expressed in chicken lung and spleen tissues. Although highly conserved among the species, chNup210 was evolutionary clustered in the same clade as that of duck compared to other mammals. Furthermore, this study revealed that chNup210 is expressed in TLR3 signaling pathway and provides fundamental information on Nup210 expression in chicken. Future studies that offer insight into the involvement of chNup210 in the chicken innate immune response against viral infection are recommended.

Different Cytokine Dependency of Proneural to Mesenchymal Glioma Stem Cell Transition in Tumor Microenvironments (종양미세환경에서 이질적인 사이토카인에 의한 PN-MES 뇌종양줄기세포 전이 조절)

  • Lee, Seon Yong;Kim, Hyunggee
    • Journal of Life Science
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    • v.29 no.5
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    • pp.530-536
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    • 2019
  • Glioblastoma (GBM) is the most incurable brain cancer derived from the transformed glial cells. Standard anti-GBM treatment, including surgery and chemoradiotherapy, does not ensure good prognosis for the patients with GBM, because successful therapy is often impeded by presence of glioma stem cells (GSCs). GSCs, which is generally divided into proneural (PN) and mesenchymal (MES) subtype, are understood as subpopulation of cancer cells responsible for GBM initiation, progression and recurrence after standard treatments. In the present study, we demonstrate that PN subtype GSCs differentially transit to MES subtype GSCs by specific cytokines. The expression of CD44, a marker of MES subtype GSCs, was observed when GSC11 PN subtype GSCs were exposed to tumor necrosis factor alpha ($TNF-{\alpha}$) cytokine and GSC23 PN subtype GSCs were treated to transforming growth factor beta 1 ($TGF-{\beta}1$) cytokine. Ivy glioblastoma atlas project (Ivy GAP) bioinformatics database showed that $TNF-{\alpha}$ and $TGF-{\beta}1$ were highly expressed in necrotic region and perivascular region, respectively. In addition, $TNF-{\alpha}$ signaling was relatively upregulated in necrotic region, while $TGF-{\beta}$ signaling was increased in perivascular region. Taken together, our observations suggest that MES subtype GSCs can be derived from various PN subtype GSCs by multimodal cytokine stimuli provided by neighboring tumor microenvironment.

The development of the 2020 Dietary Reference Intakes for Korean population: Lessons and challenges (2020 한국인 영양소 섭취기준 제·개정: 교훈과 도전)

  • Kwon, Oran;Kim, Hyesook;Kim, Jeongseon;Hwang, Ji-Yun;Lee, Jounghee;Yoon, Mi Ock
    • Journal of Nutrition and Health
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    • v.54 no.5
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    • pp.425-434
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    • 2021
  • The discovery of the relationship between nutrients and deficiency diseases during the 100 years from the mid-1800s to the mid-1900s was a breakthrough that led to advances in the study of nutrition. The Recommended Dietary Allowances (RDA) were created as a quantitative standard for avoiding diseases caused by nutrient deficiency. In addition, a reductionism paradigm has become generally accepted among nutrition scholars in health and disease, which focused on the properties of individual nutrients, content in foods, cellular levels, and mechanisms of action. The reductionist paradigm worked very well for the prevention and treatment of malnutrition diseases. However, as the incidence of nutrient deficiencies decreased and that of chronic diseases increased, the nutrition goals have been changed to secure safe and adequate nutrient intake and to reduce chronic disease risks. Accordingly, Dietary Reference Intakes (DRIs), a set of nutrient-based reference values, were designed to replace the RDA. The revised Korean DRIs were published for 40 nutrients in 2020. However, there is still room for improvement in the reference intake levels targeted at reducing the risk of chronic disease. The reductionist approach can no longer be practical because chronic diseases are related to the interactions between multi-components in the foods and multi-targets in the body. Therefore, a second innovative leap is needed following the nutrition development breakthrough made over 100 years ago. To this end, the nutrition paradigm must evolve from reductionism to a holism approach. Cutting-edge scientific technologies, such as metabolomics, transcriptomics, microbiomics, and bioinformatics, should also be acceptable in nutrition science based on the knowledge gained from basic nutrition studies.