• Title/Summary/Keyword: Cell-Counting

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Immunomodulatory effects of fermented Platycodon grandiflorum extract through NF-κB signaling in RAW 264.7 cells

  • Park, Eun-Jung;Lee, Hae-Jeung
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.453-462
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    • 2020
  • BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

Effect of Shear Stress on Bovine Aortic Smooth Muscle Cell Growth (우 대동맥 평활근 세포의 성장에 관한 shear stress의 영향)

  • 김동욱
    • KSBB Journal
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    • v.11 no.1
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    • pp.52-57
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    • 1996
  • Bovine aortic smooth muscle cells cultured on the slide glass were exposed to sheared flow up to 120 hours in flow chamber to see the effect of shear stress on cell growth in wall shear stresses of 0 to 26dyn/$cm^2$. From lactate dehydrogenase concentration measurement of the circulating medium, it was shown that sheared flow in the shear stress range did not remove additional smooth muscle cells from the slide glass compared with cells in stationary condition. According to smooth muscle cell counting per$cm^2$ of the surface, smooth muscle cells grew fastest in the stationary condition. As the wall shear stress increased, the growth of cells became slower. When the wall shear stress increased over 17dyn/$cm^2$, cell growth was not observed throughout the experiment.

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Alternation in the Cell-mediated Immune Reaction by Bisphenol A Exposure in vitro (In vitro에서 Bisphenol A 노출에 의한 세포성 면역반응의 변화)

  • Pyo Myoung-Yun
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.92-96
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    • 2005
  • In order to investigate the effects of bisphenol A (BPA) on cell mediated immune reaction in vitro we examined the allogenic mixed lymphocyte reaction (MLR), splenocytes proliferation (SP) to T cell mitogens and IFN-${\gamma}\;production$. Splenocytes of Balb/c mice ($1.5{\times}10^5$ cells/well) were co-cultured with different numbers of mitomycin C-treated mature dentritic cells (DCs) in presence of BPA (25, 50, 100 ${\mu}M$) and $[^{3}H]$thymidine incorporation (cpm) was measured by scintilation counting. Splenocytes ($2{\times}10^6$ cells/well) were cultured with mitogens, Con A ($2\;{\mu}g/ml$), PHA ($5\;{\mu}g/ml$) and IL-2 ($0.1\;{\mu}g/ml$), or PMA ($5\;{\mu}g/ml$) and INO ($1\;{\mu}g/ml$) in presence of BPA (1, 10, 25, 50, 100 ${\mu}M$) and SP was assessed by MTT assay. $IFN-{\gamma}$ levels in culture supernant were determined by ELISA. At low concentration, BPA slightly increased MLR, SP and $IFN-{\gamma}$ levels, but at higher concentration it showed significant inhibitory effects on these immunological parameters. These results indicate that BPA is able to alternate cell-mediated immune reaction.

Study on Cytotoxic Activities of the Essential Oil Compounds from Ligusticum chuanxiong against Some Human Cancer Strains (토천궁 정유 성분의 수종 사람 암 세포주에 대한 세포 독성)

  • Sim, Youn;Shin, Seung-Won
    • YAKHAK HOEJI
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    • v.55 no.5
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    • pp.398-403
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    • 2011
  • Ligusticum chuanxiong (Umbelliferae) is a perennial herb that has been used for invigoration of blood in Korean traditional medicine. It is especially important in gynecological therapy of amenorrhea and dysmenorrhea. In this study, the essential oil of L. chuanxiong was obtained by steam distillation and its main components of L. chuanxiong, Z-ligustilide and butylidene phthalide, were isolated by silica gel column chromatography. We investigated the cytotoxic effects of the essential oil fraction of L. chuanxiong and its main components on MCF-7, HeLa and SK-Hep-1 cell lines by measuring the number of surviving cancer cells after treatment through direct cell counting and MTT analysis, and by examining the morphological changes under the microscope. The essential oil from the rhizomes of L. chuanxiong and its main components showed significant cytotoxic activities for all three tested cell lines. We also observed morphological changes of shrinking and blebbing in the membranes of the three cell lines, depending on the concentration of L. chaunxiong oil or its main components.

Effects of Hominis placenta pharmacopuncture on the blood picture and antioxidative activity in rats (자하거(紫河車)약침이 흰쥐의 혈액성상과 항산화효소의 활성에 미치는 영향)

  • Lee, Joon-Moo
    • Korean Journal of Acupuncture
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    • v.26 no.2
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    • pp.53-60
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    • 2009
  • Objectives: To investigate the effects of Hominis placenta pharmacopuncture on the blood picture and antioxidative activity in rats. Methods: Sprague-Dawley rats were divided into 3 groups; normal control (n=5), pharmacopuncture at CV12 (CV12 group, n=5), and pharmacopuncture at ST36 (ST36 group, n=5) once every other day for 4 weeks. Blood cell counting was performed and liver superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) activities were analyzed. Results: Values of red blood cell and plasma cell volume were significantly higher in the ST36 group than the normal control. Values of hematocrit, total protein, and albumin were not significantly different among groups. White blood cell count and the percentage of neutrophils, lymphocytes, and eosinophils were not significantly different among groups. However, monocytes and basophils were significantly increased in the ST36, and CV12 groups, respectively. SOD and CAT in the CV12 and ST36 groups were significantly activated than in the normal control group, while the activity of GSH-Px showed no significant difference among groups. Conclusions: Based on the findings of this study, Hominis placenta pharmacopuncture may have positive impact on antioxidative capacity, thus activate various functions of the body.

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Acceleration of Cell Proliferation and Gene Expression in Human Chondrosarcoma Cells Stimulated by Strong Pulse Magnetic Field

  • Shin, Sung Chul;Chung, Eui Ryong;Hwang, Do Guwn
    • Journal of Magnetics
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    • v.18 no.1
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    • pp.14-20
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    • 2013
  • For the treatment of osteoarthritis, pulsed electromagnetic field stimulus has been suggested as a useful therapeutic method in rehabilitative medicine. Most studies have been performed under low-frequency and low-energy to find out biological properties for stimulating chondrocyte with pulsed magnetic field. In this study, the effect of strong pulse magnetic field on the human chondrosarcoma cells (SW-1353) has been investigated by means of cell counting, morphologies, and gene expression of cartilage extracellular matrix genes. The SW-1353 cells were exposed under the field intensities of 270, 100, 55, 36, and 26 mTesla during 6 hours a day in 5 consecutive days. The pulse magnetic field with an LRC oscillating signal has the pulse width of 0.126 msec and stimulation period of 1 sec. For the 270 and 100 mTesla stimulation, the cell proliferation significantly increased in 21-24% as compared with the non-stimulated cells. Gene expression of cartilage extracellular matrix genes (ACAN, COMP and COL2A1) was assayed by quantitative real time-PCR method. The ACAN gene expression showed a significant brightness, which means the increase on gene expression, compared with the non-stimulated cells. Our results suggest that the strong pulse magnetic field stimulation can be utilized to accelerate cell proliferation and gene expression on human chondrosarcoma cells.

Thymol and eugenol in essential oils enhance phage endolysin LysECP26-mediated cell wall disruption of Escherichia coli O157:H7

  • Park, Do-Won;Lee, Jong Hun;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.53 no.6
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    • pp.819-822
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    • 2021
  • To enhance phage endolysin-mediated cell wall disruption of Escherichia coli O157:H7, the cells were co-treated with aromatic compounds, namely thymol or eugenol, found in essential oils and endolysin LysECP26. Interestingly, the minimal inhibitory concentrations of LysECP26 was four times lower when used in combination with either of the two compounds than when it was used alone. This synergistic activity was also confirmed by viable cell counting. Within 1 h of LysECP26 and eugenol or thymol co-treatment to the cells, there was a 2.3 or 3.8 log CFU/mL reductions, respectively. Additionally, field emission scanning electron microscopy showed cell wall disruption and severe morphological alterations of the cells in case of the combination treatments. Therefore, endolysin and thymol or eugenol co-treatment can help in developing efficient bio-control strategies against gram-negative pathogen E. coli O157:H7.

Suppression of MCF-7 Human Breast Cancer Cell Proliferation by Globefish Takifugu obscurus Homogenate (복어(Takifugu obscurus) 균질액에 의한 MCF-7 인간 유방암세포 성장 억제 효과)

  • Kim, Junghoon;Kim, Jungho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.6
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    • pp.878-885
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    • 2020
  • Previously, we reported that globefish Takifugu obscurus homogenate suppresses the growth of human colorectal cancer cells. To extend the applications of globefish homogenate, we investigated its cytotoxic effects on human breast cancer cells. To assess the effects of globefish homogenate on growth of MCF (Michigan Cancer Foundation)-7 human breast cancer cells, cell proliferation and colony formation assays were performed using the cell counting and Crystal Violet staining methods. The 50% inhibitory concentration (IC50) of globefish homogenate on MCF-7 cell proliferation was calculated from the sigmoidal dose-response curve. The colony formation assay demonstrated that MCF-7 cells treated with globefish homogenate formed up to 80% fewer colonies than control MCF-7 cells. Treatment with globefish homogenate markedly suppressed the growth of MCF-7 cells in a dose-dependent manner. The sensitivity of the cells to globefish homogenate was determined by calculating the IC50; in this case, the IC50 was 210 ㎍/mL. Furthermore, significant downregulation of Cyclin D1 expression, along with phospho-Akt and total Akt levels, was observed in MCF-7 cells treated with globefish homogenate. This study demonstrates that treatment with globefish homogenate inhibits the proliferation of MCF-7 human breast cancer cells by downregulating the expression of phosphor-Akt, total Akt, and Cyclin D1 proteins.

Development of microfluidic green algae cell counter based on deep learning (딥러닝 기반 녹조 세포 계수 미세 유체 기기 개발)

  • Cho, Seongsu;Shin, Seonghun;Sim, Jaemin;Lee, Jinkee
    • Journal of the Korean Society of Visualization
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    • v.19 no.2
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    • pp.41-47
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    • 2021
  • River and stream are the important water supply source in our lives. Eutrophication causes excessive green algae growth including microcystis, which makes harmful to ecosystem and human health. Therefore, the water purification process to remove green algae is essential. In Korea, green algae alarm system exists depending on the concentration of green algae cells in river or stream. To maintain the growth amount under control, green algae monitoring system is being used. However, the unmanned, small and automatic monitoring system would be preferable. In this study, we developed the 3D printed device to measure the concentration of green algae cell using microfluidic droplet generator and deep learning. Deep learning network was trained by using transfer learning through pre-trained deep learning network. This newly developed microfluidic cell counter has sufficient accuracy to be possibly applicable to green algae alarm system.

Quantification of the ichthyotoxic raphidophyte Chattonella marina complex by applying a droplet digital PCR

  • Juhee, Min;Kwang Young, Kim
    • ALGAE
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    • v.37 no.4
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    • pp.281-291
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    • 2022
  • Quantifying the abundance of Chattonella species is necessary to effectively manage the threats from ichthyotoxic raphidophytes, which can cause large-scale mortality of aquacultured fish in temperate waters. The identification and cell counting of Chattonella species have been conducted primarily on living cells without fixation by light microscopy because routine fixatives do not retain their morphological features. Species belonging to the Chattonella marina complex, including C. marina and C. marina var. ovata, had high genetic similarities and the lack of clear morphological delimitations between the species. To estimate the abundance of C. marina complex in marine plankton samples, we developed a protocol based on the droplet digital polymerase chain reaction (ddPCR) assay, with C. marina complex-specific primers targeting the internal transcribed spacer (ITS) region of the rDNA. Cell abundance of the C. marina complex can be determined using the ITS copy number per cell, ranging from 25 ± 1 for C. marina to 112 ± 7 for C. marina var. ovata. There were no significant differences in ITS copies estimated by the ddPCR assay between environmental DNA samples from various localities spiked with the same number of cells of culture strains. This approach can be employed to improve the monitoring efficiency of various marine protists and to support the implementation of management for harmful algal blooms, which are difficult to analyze using microscopy alone.