• Title/Summary/Keyword: single cells

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Application of Single Cell Gel Electrophoresis for Detection of DNA Single Strand Breaks in DNA of Fish Blood Cell (어류혈구세포에 있어서 Single Cell Gel Electrophoresis를 응용한 DNA Single Strand Breack의 측정)

  • KIM Gi Beum;LEE Richard F.;MARUYA Keith A.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.36 no.4
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    • pp.346-351
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    • 2003
  • Single-cell gel electrophoresis (comet assay) was used to detect DNA single strand break in blood cells from several marine fish species. Three fish species were collected from Georgia coastal area. Mummichog, Fundulus heteroclitus showed higher DNA damage than sea bass, Lateolabrax japonicus and trout, Oncorhynchus masou masou under the same experimental conditions. Mummichogs had more alkaline-labile sites on their DNA than other fish species. The comet assay with mummichog blood cells at pH 12.5 showed a dose-response curve with the increasing concentrations of hydrogen peroxide. While the isolated leucocytes showed no increase of DNA damage after in vitro exposure to 2-methyl-1,4-naphthoquinone (MNQ), erythrocytes showed dose-dependent DNA damage. These results indicate that the comet assay can be applied successfully as a bioassay using erythrocyte for environmental monitoring.

Single-Cell Genomics for Investigating Pathogenesis of Inflammatory Diseases

  • Seyoung Jung;Jeong Seok Lee
    • Molecules and Cells
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    • v.46 no.2
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    • pp.120-129
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    • 2023
  • Recent technical advances have enabled unbiased transcriptomic and epigenetic analysis of each cell, known as "single-cell analysis". Single-cell analysis has a variety of technical approaches to investigate the state of each cell, including mRNA levels (transcriptome), the immune repertoire (immune repertoire analysis), cell surface proteins (surface proteome analysis), chromatin accessibility (epigenome), and accordance with genome variants (eQTLs; expression quantitative trait loci). As an effective tool for investigating robust immune responses in coronavirus disease 2019 (COVID-19), many researchers performed single-cell analysis to capture the diverse, unbiased immune cell activation and differentiation. Despite challenges elucidating the complicated immune microenvironments of chronic inflammatory diseases using existing experimental methods, it is now possible to capture the simultaneous immune features of different cell types across inflamed tissues using various single-cell tools. In this review, we introduce patient-based and experimental mouse model research utilizing single-cell analyses in the field of chronic inflammatory diseases, as well as multi-organ atlas targeting immune cells.

DNA Damage and Micronuclei Induced by Di (2-ethylhexyl) phthalate in Human Breast Carcinoma MCF-7 cells (Di(2-ethylhexyl) phthalate에 의해 유도된 DNA손상과 소핵 형성)

  • 김종원;한의식;박미선;엄미옥;김인숙;전혜승;정해관;심웅섭;오혜영
    • Environmental Mutagens and Carcinogens
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    • v.21 no.1
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    • pp.34-43
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    • 2001
  • Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCE-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCE-10A cells, and female peripheral blood cells using micronucleus assay and in human breast carcinoma MCF-7 cells up to $1.28$\times$10^{-2}$ M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCE-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.

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An Anatomical and Histochemical Study of the Olfactory Organ in Rice-fish Oryzias sinensis(Pisces: Adrianichthyidae) in South Korea (한국산 대륙송사리 Oryzias sinensis(Pisces: Adrianichthyidae) 후각기관의 해부 및 조직화학적 연구)

  • Kim, Hyun Tae;Lee, Yong Joo;Park, Jong Young
    • Korean Journal of Ichthyology
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    • v.28 no.4
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    • pp.223-228
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    • 2016
  • The anatomy and histology of the olfactory organ in Oryzias sinensis was researched using a stereo microscope and light microscope. In the gross structure, the paired olfactory organs on the dorsal part of the head consist of two nostrils (a circular anterior nostril and a slit posterior nostril in a distance), a single olfactory chamber and a single accessory nasal sac. In the histological study, the epithelium of the olfactory chamber is classified into both sensory and non-sensory regions. The sensory epithelium consists of olfactory receptor neurons, supporting cells, basal cells and vesicles, and is islet in distributional pattern. The non-sensory epithelium is composed of stratified epithelial cells and two types of mucous cells (acidic and neutral cells). The epithelium of the accessory nasal sac has swollen stratified epithelial cells, mucous cells with a rich glycoprotein. Such an olfactory anatomy and histology of O. sinensis may reflect its habitat surrounding stagnant and polluted water.

An Efficient SLC Transition Method for Improving Defect Rate and Longer Lifetime on Flash Memory (플래시 메모리 상에서 불량률 개선 및 수명 연장을 위한 효율적인 단일 비트 셀 전환 기법)

  • Hyun-Seob Lee
    • Journal of Internet of Things and Convergence
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    • v.9 no.3
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    • pp.81-86
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    • 2023
  • SSD (solid state disk), which is flash memory-based storage device, has the advantages of high density and fast data processing. Therefore, it is being utilized as a storage device for high-capacity data storage systems that manage rapidly increasing big data. However, flash memory, a storage media, has a physical limitation that when the write/erase operation is repeated more than a certain number of times, the cells are worn out and can no longer be used. In this paper, we propose a method for converting defective multi-bit cells into single-bit cells to reduce the defect rate of flash memory and extend its lifetime. The proposed idea distinguishes the defects and treatment methods of multi-bit cells and single-bit cells, which have different physical characteristics but are treated as the same defect, and converts the expected defective multi-bit cells into single-bit cells to improve the defect rate and extend the overall lifetime. Finally, we demonstrate the effectiveness of our proposed idea by measuring the increased lifetime of SSD through simulations.

Single-cell RNA sequencing reveals the heterogeneity of adipose tissue-derived mesenchymal stem cells under chondrogenic induction

  • Jeewan Chun;Ji-Hoi Moon;Kyu Hwan Kwack;Eun-Young Jang;Saebyeol Lee;Hak Kyun Kim;Jae-Hyung Lee
    • BMB Reports
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    • v.57 no.5
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    • pp.232-237
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    • 2024
  • This study investigated how adipose tissue-derived mesenchymal stem cells (AT-MSCs) respond to chondrogenic induction using droplet-based single-cell RNA sequencing (scRNA-seq). We analyzed 37,219 high-quality transcripts from control cells and cells induced for 1 week (1W) and 2 weeks (2W). Four distinct cell clusters (0-3), undetectable by bulk analysis, exhibited varying proportions. Cluster 1 dominated in control and 1W cells, whereas clusters (3, 2, and 0) exclusively dominated in control, 1W, and 2W cells, respectively. Furthermore, heterogeneous chondrogenic markers expression within clusters emerged. Gene ontology (GO) enrichment analysis of differentially expressed genes unveiled cluster-specific variations in key biological processes (BP): (1) Cluster 1 exhibited up-regulation of GO-BP terms related to ribosome biogenesis and translational control, crucial for maintaining stem cell properties and homeostasis; (2) Additionally, cluster 1 showed up-regulation of GO-BP terms associated with mitochondrial oxidative metabolism; (3) Cluster 3 displayed up-regulation of GO-BP terms related to cell proliferation; (4) Clusters 0 and 2 demonstrated similar up-regulation of GO-BP terms linked to collagen fibril organization and supramolecular fiber organization. However, only cluster 0 showed a significant decrease in GO-BP terms related to ribosome production, implying a potential correlation between ribosome regulation and the differentiation stages of AT-MSCs. Overall, our findings highlight heterogeneous cell clusters with varying balances between proliferation and differentiation before, and after, chondrogenic stimulation. This provides enhanced insights into the single-cell dynamics of AT-MSCs during chondrogenic differentiation.

Adenovirus-mediated Double Suicide Gene Selectively Kills Gastric Cancer Cells

  • Luo, Xian-Run;Li, Jian-Sheng;Niu, Ying;Miao, Li
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.781-784
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    • 2012
  • The aim of this study was to evaluate the effect of the adenovirus-mediated double suicide gene (CD/TK) for selective killing of gastric cancer cells. Gastric cancer cells SCG7901 and normal gastric epithelial cell lines were infected by adenoviruses Ad-survivin/GFP and Ad-survivin/CD/TK. GFP expression and CD-TK were detected by fluorescence microscopy and reverse transcriptase polymerase chain reaction (RT-PCR), respectively. After treatment of the infected cells with the pro-drugs ganciclovir (GCV) and/or 5-FC, the cell growth status was evaluated by methyl thiazolyl tetrazolium assay. Cell cycle changes were detected using flow cytometry. In nude mice bearing human gastric cancer, the recombinant adenovirus vector was injected directly into the tumor followed by an intraperitoneal injection of GCV and/or 5-FC. The subsequent tumor growth was then observed. The GFP gene driven by survivin could be expressed within the gastric cancer line SCG7901, but not in normal gastric epithelial cells. RT-PCR demonstrated the presence of the CD/TK gene product in the infected SCG7901 cells, but not in the infected normal gastric epithelial cells. The infected gastric cancer SCG7901, but not the gastric cells, was highly sensitive to the pro-drugs. The CD/TK fusion gene system showed significantly greater efficiency than either of the single suicide genes in killing the target cells (P<0.01). Treatment of the infected cells with the pro-drugs resulted in increased cell percentage in G0-Gl phase and decreased percentage in S phase. In nude mice bearing SCG7901 cells, treatment with the double suicide gene system significantly inhibited tumor growth, showing much stronger effects than either of the single suicide genes (P<0.01). The adenovirus-mediated CD/TK double suicide gene driven by survivin promoter combined with GCV an 5-FC treatment could be an effective therapy against experimental gastric cancer with much greater efficacy than the single suicide gene CD/TK combined with GCV or 5-FC.

Organic-Inorganic Perovskite for Highly Efficient Tandem Solar Cells (고효율 적층형 태양전지를 위한 유무기 페로브스카이트)

  • Park, Ik Jae;Kim, Dong Hoe
    • Ceramist
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    • v.22 no.2
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    • pp.146-169
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    • 2019
  • To overcome the theoretical efficiency of single-junction solar cells (> 30 %), tandem solar cells (or multi-junction solar cells) is considered as a strong nominee because of their excellent light utilization. Organic-inorganic halide perovskite has been regarded as a promising candidate material for next-generation tandem solar cell due to not only their excellent optoelectronic properties but also their bandgap-tune-ability and low-temperature process-possibility. As a result, they have been adopted either as a wide-bandgap top cell combined with narrow-bandgap silicon or CuInxGa(1-x)Se2 bottom cells or for all-perovskite tandem solar cells using narrow- and wide-bandgap perovskites. To successfully transition perovskite materials from for single junction to tandem, substantial efforts need to focus on fabricating the high quality wide- and narrow-bandgap perovskite materials and semi-transparent electrode/recombination layer. In this paper, we present an overview of the current research and our outlook regarding perovskite-based tandem solar technology. Several key challenges discussed are: 1) a wide-bandgap perovskite for top-cell in multi-junction tandem solar cells; 2) a narrow-bandgap perovskite for bottom-cell in all-perovskite tandem solar cells, and 3) suitable semi-transparent conducting layer for efficient electrode or recombination layer in tandem solar cells.

In Vivo Stem Cell Imaging Principles and Applications

  • Seongje Hong;Dong-Sung Lee;Geun-Woo Bae;Juhyeong Jeon;Hak Kyun Kim;Siyeon Rhee;Kyung Oh Jung
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.363-375
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    • 2023
  • Stem cells are the foundational cells for every organ and tissue in our body. Cell-based therapeutics using stem cells in regenerative medicine have received attracting attention as a possible treatment for various diseases caused by congenital defects. Stem cells such as induced pluripotent stem cells (iPSCs) as well as embryonic stem cells (ESCs), mesenchymal stem cells (MSCs), and neuroprogenitors stem cells (NSCs) have recently been studied in various ways as a cell-based therapeutic agent. When various stem cells are transplanted into a living body, they can differentiate and perform complex functions. For stem cell transplantation, it is essential to determine the suitability of the stem cell-based treatment by evaluating the origin of stem, the route of administration, in vivo bio-distribution, transplanted cell survival, function, and mobility. Currently, these various stem cells are being imaged in vivo through various molecular imaging methods. Various imaging modalities such as optical imaging, magnetic resonance imaging (MRI), ultrasound (US), positron emission tomography (PET), and single-photon emission computed tomography (SPECT) have been introduced for the application of various stem cell imaging. In this review, we discuss the principles and recent advances of in vivo molecular imaging for application of stem cell research.

Plant Regeneration from Single Cell Culture of Wheat (Triticum aestivum L.) (밀(Triticum aestivum L.)의 단세포 배양에 의한 식물체의 재분화)

  • 김시철
    • Journal of Plant Biology
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    • v.32 no.4
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    • pp.227-233
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    • 1989
  • Single cells obtained from suspension culture of mature embryo-derived callus in wheat(Triticum aestivum L. cv Jang Kwang) were cultured to regenrated into the plantlet. Cell clusters and embryogenic calluses were efficiently developed from when the single cells clutured on the MS medium supplemented with 10${\mu}{\textrm}{m}$ 2,4-D. Upon transfer to hormone-free MS medium containing 10 mg/I AgNO3, embryogenic calluses gave rise to shoots, probably through somatic embryogenesis.

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