• Title/Summary/Keyword: single cells

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Effects of Higenamine on the Calcium Current and the Action Potential in the Guinea-pig Myocytes (Higenamine이 단일심근세포의 Ca-전류 및 활동전압에 미치는 효과)

  • Kim, Young-Duck;So, In-Suk;Earm, Yung-E
    • The Korean Journal of Physiology
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    • v.21 no.2
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    • pp.169-177
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    • 1987
  • The effects of higenamine were investigated in the single atrial and ventricular myocyte of the guinea pig by using patch clamp method. The results obtained were as follows: 1) Isoprenaline which is known to be ${\beta}-agonist$ increased the duration of action potential and calcium current in ventricular cells. 2) Higenamine also increased the duration of action potential and calcium current in ventricular myocytes. And its effect was blocked by propranolol. 3) In the atrial cells, isoprenaline showed ${\beta}-agonist$ effects, which were increasing the duration of action potential and calcium current same as in ventricular cells. 4) Higenamine, however, showed the opposite effects of ${\beta}-agonist$ which were decreasing the duration of action potential and calcium current. The above results suggest that higenamine has the typical ${\beta}-agonist$ effect in ventricular cells but inhibitory effect in atrial cells and this effect on atrium could be due to the reduction of calcium current.

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Low-dose of Ultraviolet radiation-, Ethyl methanesulfonateor Bleomycin-lnduced Adaptive Response in Chinese hamster ovary Cells

  • Lee, Dong-Wook;Shin, Eun-Joo;Um, Kyung-Il
    • Environmental Mutagens and Carcinogens
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    • v.15 no.2
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    • pp.94-99
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    • 1995
  • The adaptive response and cross-adaptive response to sister chromatid exchanges (SCEs) and DNA single-strand breaks (SSBs) in Chinese hamster ovary (CHO)-K$_1$ cells treated with ultraviolet radiation (UV), ethyl methanesuffonate (EMS), or bleomycin (BLM) were investigated. Two assays were used in this study; SCEs and alkaline elution. The pretreatment with low conditioning dose of 2 mM EMS or 1 J/m$^2$ UV decreased the yield of SCEs induced by subsequent treatment with 8 mM EMS, 5 J/m$^2$ UV or 5 $\mu$g/ml BLM. And the pretreatment with low conditioning dose of 1 $\mu$g/ml BLM decreased the yield of SCEs induced by subsequent treatment with 5 $\mu$g/ml BLM or 5 J/m$^2$ UV. The rejoining of DNA SSBs in cells subsequently treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM is higher than that only treated with 2 J/m$^2$ UV, 50 mM EMS or 400 $\mu$g/ml BLM. These results suggest that there are the adaptive response and cross-adaptive response to SCEs, and is the adaptive response to the rejoining of DNA SSBs in CHO cells.

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Ultrastructure and Mrphological Fatures of Mcoplasma pneumoniae during Clture Dvelopment (Mycoplasma Pnemoniae 세포의 발달과정 중 미세구조 및 형태학적 특징)

  • Kim, Chi-Kyung;Pfister Robert M.;Somerson Norman L.
    • Korean Journal of Microbiology
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    • v.17 no.1
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    • pp.1-15
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    • 1979
  • Mycoplasma pneumoniae strain CL-s attached to broth-covered surfaces was examined sequentially during growth from single cells for morphologic and ultrastructural changes using several different electron microscopic techniques. Changes in morphology revealed both round and spindle shapes and observation of cell transitions suggested some type of morphological cycle. The round to-ovoid cells observed in the early stages of growth appeared to be viable, and morphologically and ultrastructurally different from the spherical fors which were produced during the latter stage of growth. The spindle segments were detected appeared to be structurally the same as the terminal cored structure seen in thin sections and may be a growing point or an attachment site of the cell. A tubular structure was observed in the core of the terminal structure and a microtubule-like element appeared to bridge between some spindle segments. A matrix sunstance was observed around single cells as well in the intercellular space of the colonies prepared by critical point metrical triple-layered cytoplasmic mermbranes, surfaces, of which appeared to be structurally different each other, were observed in young cells, whereas symmetrical and thicker membranes were seen in older cells. Small bodies were found in 4d or older cultures and did not appear to contain any internal structures or an easily detectable unit membrane.

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The Replication of Parvovirus KBSH DNA in the Embryonic Swine Kidney Cells (돼지 태아 신장세포에 있어 parvovirus KBSH의 DNA 복제과정에 관하여)

  • 이성욱;김연수;강현삼
    • Korean Journal of Microbiology
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    • v.25 no.1
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    • pp.34-39
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    • 1987
  • To study the replication process of the single-stranded DNA parvovirus KBSH-isolated from normal human cell cultures-in actively dividing embryonic swine kidney cells, amount of the synthesized viral hemagglutination (HA) antigen and the overall rate of viral double-stranded replicative form(RF) DNA synthesis were wxamined. The initiation of viral RF KNA synthesis and the decrease of host DNA synthesis rate in viral infected cells occurred almost same time at 15-16 hour post infection(PI). And the release of viral HA antigen to media followed at 24 hour PI, concurrently the overall rate of viral RF DNA synthesis reaching its maximum. Evidence is presented which indicates that successful performance of viral RF DNA replication requires proteins synthesized in viral infected cells at 10-14 hour PI.

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Transcriptional Heterogeneity of Cellular Senescence in Cancer

  • Junaid, Muhammad;Lee, Aejin;Kim, Jaehyung;Park, Tae Jun;Lim, Su Bin
    • Molecules and Cells
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    • v.45 no.9
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    • pp.610-619
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    • 2022
  • Cellular senescence plays a paradoxical role in tumorigenesis through the expression of diverse senescence-associated (SA) secretory phenotypes (SASPs). The heterogeneity of SA gene expression in cancer cells not only promotes cancer stemness but also protects these cells from chemotherapy. Despite the potential correlation between cancer and SA biomarkers, many transcriptional changes across distinct cell populations remain largely unknown. During the past decade, single-cell RNA sequencing (scRNA-seq) technologies have emerged as powerful experimental and analytical tools to dissect such diverse senescence-derived transcriptional changes. Here, we review the recent sequencing efforts that successfully characterized scRNA-seq data obtained from diverse cancer cells and elucidated the role of senescent cells in tumor malignancy. We further highlight the functional implications of SA genes expressed specifically in cancer and stromal cell populations in the tumor microenvironment. Translational research leveraging scRNA-seq profiling of SA genes will facilitate the identification of novel expression patterns underlying cancer susceptibility, providing new therapeutic opportunities in the era of precision medicine.

Anti-tumor Effect of 4-1BBL Modified Tumor Cells as Preventive and Therapeutic Vaccine

  • Hong Sung Kim
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.312-316
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    • 2022
  • We have previously reported that genetically modified tumor cells with 4-1BBL have anti-cancer effects in a CT26 mouse colorectal tumor model. In this study, genetically modified tumor cells with 4-1BBL were evaluated for their potential as candidates for preventive and therapeutic cancer vaccine. To identify the effect of preventive and therapeutic vaccine of genetically modified tumor cells with 4-1BBL, tumor growth pattern of CT26-4-1BBL as a cancer vaccine was examined compared to CT26-beta-gal. In therapeutic vaccination, CT26-WT was inoculated into mice and then vaccinated mice with doxorubicin (Dox)-treated CT26-beta-gal and CT26-4-1BBL (single or three times). Triple vaccination with Dox-treated tumor cell inhibited tumor growth compared to single vaccination. Vaccination with CT26-4-1BBL showed an efficient tumor growth inhibition compared to vaccination with CT26-beta-gal. For preventive vaccination, Dox-treated CT26-beta-gal and CT26-4-1BBL was vaccinated into mice with three times and then administered mice with CT26-WT. Preventive vaccination with CT26-4-1BBL showed no tumor growth. Preventive vaccination with CT26-beta-gal also led to tumor-free mice. These results suggest that genetically modified tumor cells with 4-1BBL can be used as therapeutic or preventive cancer vaccine.

Optimization of DNA Extraction from a Single Living Ciliate for Stable and Repetitive PCR Amplification

  • Kim, Se-Joo;Min, Gi-Sik
    • Animal cells and systems
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    • v.13 no.3
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    • pp.351-356
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    • 2009
  • Ciliates are undoubtedly one of the most diverse protozoans that play a significant role in ecology. However, molecular examination, based on comparing the DNA sequences, has been done on a limited number of the species. Because most ciliates are uncultivable and their population sizes are often too small, it is usually difficult to obtain sufficient genomic DNA required for PCR based experiments. In the present study, we evaluated the effectiveness of four commercial DNA extraction procedures that extract high quality genomic DNA from a single ciliate cell. It was discovered that RED Extract-N-$Amp^{TM}$ PCR kit is the best method for removing PCR-inhibiting substances and minimizing DNA loss during purification. This method can also amplify more than 25 reactions of PCR. In addition, this technique was applied to single cells of 19 species belonged to 7 orders under 5 classes that isolated from mixed natural populations. Their small subunit ribosomal DNA (SSU rDNA) was successfully amplified. In summary, we developed a simple technique for the high-yield extraction of purified DNA from a single ciliate cell that may be more useful for rare ciliates, such as tiny and uncultivable marine microbes.

Expression of Cyclin B1 mRNA and Protein after Activation in Enucleated Mouse Oocytes

  • Hwang, Seong-Soo;Kim, Chang-Kun;Chung, Young-Chai
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.116-116
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    • 2002
  • Further development of reconstructed embryos may be dependent upon the synchronization of donor nucleus and recipient cytoplasm at cell fusion, To control the synchronization of donor and recipient cells, the enucleated MII arrested oocytes are artificially stimulated prior to embryo reconstruction. Destruction of cyclin B results in the exit of cells from M-phase of cell cycle. This study was designed to investigate the effects of single or combined stimulation affected cyclin B1 mRNA and protein levels in mouse oocytes. The oocyte activation was induced by 7% ethanol or 10$\mu\textrm{g}$/$m\ell$ Ca-ionophore without (single) or with (combined) 10$\mu\textrm{g}$/$m\ell$ cycloheximide. Competitive quantitative PCR for cyclin Bl mRNA and western blot analysis for cyclin B1 protein was preformed in mouse oocytes. Cyclin B1 mRNA level was significantly reduced in single (P<0.05) and combined (P<0.05) stimulation groups. However, this level did not change in non-activated group and increased in intact group. Cyclin B1 protein level was also significantly reduced in both single (P<0.05) and combined (P<0.05) stimulation groups. In conclusion, single and combined stimulation induces the degradation of cyclin B1 mRNA and protein after activation in enucleated mouse oocytes.

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Recyclable single-stranded DNA template for synthesis of siRNAs

  • Ali, Mussa M.;Obregon, Demian;Agrawal, Krishna C.;Mansour, Mahmoud;Abdel-Mageed, Asim B.
    • BMB Reports
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    • v.43 no.11
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    • pp.732-737
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    • 2010
  • RNA interference is a post-transcriptional silencing mechanism triggered by the bioavailability and/or exogenous introduction of double-stranded RNA (dsRNA) into cells. Here we describe a novel method for the synthesis of siRNA in a single vessel. The method employs in vitro transcription and a single-stranded DNA (ssDNA) template and design, which incorporates upon self-annealing, two promoters, two templates, and three loop regions. Using this method of synthesis we generated efficacious siRNAs designed to silence both exogenous and endogenous genes in mammalian cells. Due to its unique design the single-stranded template is easily amenable to adaptation for attachment to surface platforms for synthesis of siRNAs. A siRNA synthesis platform was generated using a 3' end-biotinylated ssDNA template tethered to a streptavidin coated surface that generates stable siRNAs under multiple cycles of production. Together these data demonstrate a unique and robust method for scalable siRNA synthesis with potential application in RNAi-based array systems.

Fabrication of Anode-Supported SOFC Single Cells via Tape-Casting of Thin Tapes and Co-Firing (박막 테이프캐스팅과 동시소성에 의한 연료극 지지형 SOFC 단전지 제조)

  • Moon, Hwan;Kim, Sun-Dong;Hyun, Sang-Hoon;Kim, Ho-Sung
    • Journal of the Korean Ceramic Society
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    • v.43 no.12 s.295
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    • pp.788-797
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    • 2006
  • An anode-supported SOFC single cell having $5{\mu}m$ thin electrolyte was fabricated cost-effectively by tape casting, laminating, and co-filing of anode (NiO-YSZ), cathode (LSM-YSZ), and electrolyte (YSZ) components. The optimal slurry compositions of the green tapes for SOFC components were determined by an analysis of the mean diameter, the slurry viscosity, the tensile strength/strain of the green tapes, and their green microstructures. The single cells with a dense electrolyte and porous electrodes could be co-fired successfully at $1325\sim1350^{\circ}C$ by controlling the contents of pore former and the ratio of coarse YSZ and fine YSZ in the anode and the cathode. The single cell co-fired at $1350^{\circ}C$ showed $100.2mWcm^{-2}$ of maximum power density at $800^{\circ}C$ but it was impossible to apply it to operate at low temperature because of low performance and high ASR, which were attributed to formation of the secondary phases in the cathode and the interface between the electrolyte and the cathode.