• Title/Summary/Keyword: flow cytometry

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Apoptosis Induction in Human Leukemic Promyelocytic HL-60 and Monocytic U937 Cell Lines by Goniothalamin

  • Petsophonsakul, Ploingarm;Pompimon, Wilart;Banjerdpongchai, Ratana
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.5
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    • pp.2885-2889
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    • 2013
  • Goniothalamin is an active compound extracted from Goniothalamus griffithii, a local plant found in northern Thailand. Goniothalamin inhibits cancer cell growth but is also toxic to normal cells. The aims of this study were to identify the cytotoxic effect of goniothalamin and the mechanism of cell death in human HL-60 and U937 cells. Cytotoxicity was determined by MTT assay and cell cycle profiles were demonstrated by staining with propidium iodide (PI) and flow cytometry. Apoptosis was confirmed by staining with annexin V-FITC/propidium iodide (PI) and flow cytometry. Reduction of mitochondrial transmembrane potential was determined by staining with dihexyloxacarbocyanine iodide and flow cytometry and expression of Smac, caspase-8 and -9 was demonstrated by Western blotting. Goniothalamin inhibited growth of HL-60 and U937 cell lines. An increase of SubG1 phase was found in their cell cycle profiles, indicating apoptosis as the mode of cell death. Apoptosis was confirmed by the flip-flop of phosphatidylserine using annexin V-FITC/PI assay in HL60 and U937 cells in a dose response manner. Furthermore, reduction of mitochondrial transmembrane potential was found in both cell types while expression of caspase-8, -9 and Smac/Diablo was increased in HL-60 cells. Taken together, our results indicate that goniothalamin-treated human leukemic cells undergo apoptosis via intrinsic and extrinsic pathways.

Flow Cytometric Assessment of Immune Parameters of the Manila Clam (Ruditapes philippinarum) (유세포 분석기를 이용한 바지락(Ruditapes philippinarum)의 면역력 측정)

  • Park Kyung-Il;Park Heung-Sik;Kim Jong-Man;Park Young-Je;Hong Jae-Sang;Choi Kwang-Sik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.39 no.spc1
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    • pp.123-131
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    • 2006
  • Immmune parameters of the Manila clam Ruditapes philippinarum collected from four tidal flats, Nudong, Gonam, Hwangdo and Bangpo on Anmyeon-do, Korea were optimized and evaluated at the single cell level using flow-cytometry Hemocytes were withdrawn from the sinus of each clam, and total hemocyte counts (THC), phagocytosis rate, hemocyte mortality (HM) and DNA damage of hemocyte were analyzed. The highest hemocyte counts was recorded from the clams collected from Gonam, followed by Hwangdo, Nudong and Bangpo (P<0.001). Phagocytosis rate and hemocyte mortality of Gonam and Nudong clams were significantly higher than those of clams from Hwangdo and Bangpo (P<0.001). DNA damage in the clams from Nudong was higher twice than that of clams from Gonam (P<0.05). We suggest that the flow-cytometry has a high potential for evaluation of immunity of marine bivalves.

Flow cytometry of cell-cycle on Flavin mononucleotide (1,4-butanediamine) Pt(II) Complex and Cisplatin and Their Biochemical Analysis of Nephrotoxicity in ICR Mice (Flavin mononucleotide (1,4-butanediamine) Pt(II) Complex와 Cisplatin의 세포주기에 대한 유세포 분석 및 ICR계 생쥐에서의 신장독성에 대한 생화학적 분석)

  • 권영이;황규자;김안근;김국환;김원규;안동춘
    • YAKHAK HOEJI
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    • v.44 no.2
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    • pp.149-154
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    • 2000
  • Flavin mononucleotide (1,4-butanediamine) Pt(II) complex (7FMN) was synthesized and screened anticancer activity [J. Pharm. Soc. Korea 43(6),762-770 (1999)]. 7FMN have good water solubility and moderate anticancer activiy In this paper cell-cycle specificity and nephrotoxicity were studied. Interaction of DNA with cisplatin and synthesized 7FMN was analyzed by flow cytometry and showed G2 arrest in L1210 cell line. It means that cell-cycle on L1210 was inhibit in S phase by cisplatin and 7FMN. In order to biochemically analyze nephrotoxicity of cisplatin and 7FMN, after injecting each agent intraperitoneally, blood was exsanguinated after 6 hours, 1 day, 3 days and 7 days, respectively. Then, serum was separated from the blood. The serum level of BUN, creatinine and uric acid in cisplatin and 7FMN administated mice (25~35 g, ICR strain, a dose each 8,12 and 16 times of the $IC_{50}$/ value, cisplatin; 7 times) were determined by autochemistry analyzer. In cisplatinadministered mice group, BUN level was elevated than normal control group at 3rd day and repaired at 7th day. In 7FMN administrated group was not elevated. Creatinine and uric acid level were no difference with the normal control group. Therefore synthesized 7FMN is less toxic than cisplatin in nephrotoxiciaty.

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Assessment of Inactivation for Salmonella spp. on Chicken Meat using Confocal Laser Microscopy and Flow Cytometry (공초점 현미경 및 유세포 분류기를 이용한 계육에서의 Salmonella균 불활성화 평가)

  • Jang, Keum-Il;Chung, Duck-Hwa;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Kim, Min-Gon;Kim, Cheorl-Ho;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.290-294
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    • 2006
  • Inactivation rates of Salmonella enteritidis in vitro and in vivo were assessed using confocal microscopy and flow cytometry. S. enteritidis was inactivated with 1% (w/v) trisodium phosphate (TSP) and live cells, and inactive cells were distinguished by staining with fluorescent probe, LIVE/DEAD BacLight Bacteria Viability stain. After TSP treatment for 1 min, most of Salmonella cells changed from green (live cells) fluorescence to red (inactive cells) fluorescence, indication of effective sanitizing. Inactivation efficiency and contamination sites of S. enteritidis on chicken skin by TSP treatment were assessed using confocal laser microscopy. Precise flow cytometry histograms for viability changes of S. enteritidis. after TSP treatments were obtained. Efficiency of various sanitizer treatments on foodborne pathogens could be assessed using this method.

Aptamers as Functional Nucleic Acids: in vitro Selection and Biotechnological Applications

  • You, Kyung-Man;Lee, Sang-Hyun;Aesul Im;Lee, Sun-Bok
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.64-75
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    • 2003
  • Aptamers are functional nucleic acids that can specially bind to proteins, peptides, amino acids. nucleotides, drugs, vitamins and other organic and inorganic compounds. The aptamers are identified from random DNA or RNA libraries by a SELEX (systematic evolution of ligands by exponential amplification) process. As aptamers have the advantage, and potential ability to be released from the limitations of antibodies, they are attractive to a wide range of therapeutic and diagnostic applications. Aptamers, with a high-affinity and specificity, could fulfil molecular the recognition needs of various fields in biotechnology. In this work, we reviewed some aptamer Selection techniques, properties, medical applications of their molecules and their biotechnological applications, such as ELONA (enzyme linked oligonucleotide assay), flow cytometry, biosensors, electrophoresis, chromatography and microarrays.

Diagnostic Value of Flow Cytometric DNA Analysis in the Evaluation of Effusions (체강삼출액의 진단에 있어서 유세포분석에 의한 DNA 함량 측정의 유용성)

  • Lee, Ji-Shin;Juhang, Sang-Woo
    • The Korean Journal of Cytopathology
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    • v.8 no.1
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    • pp.20-26
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    • 1997
  • The specificity of cytologic examination in effusions is high but the sensitivity is low. Therefore, various ancillary methods for the detection of malignant cells in effusions have been proposed. The presence of an aneuploid cell population is generally considered diagnostic of malignancy. The purpose of this study is to determine whether the routine use of flow cytometry adds to standard cytologic evaluation in effusions. We did flow cytometric DNA analysis in 76 effusions(28 malignant and 48 benign fluids). All the 48 benign effusions were diploid. There were 12(42.9%) aneuploid and 16(67.1%) diploid malignant effusions. Based on these results flow cytometric DNA analysis had a sensitivity of 42.9% and a specificity of 100%. These results suggest that flow cytometric DNA analysis may be a useful adjunct to conventional cytology, but its principal limitation is us relatively low sensitivity.

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The Expression of Galectin-3, a Beta-Galactoside Binding Protein, in Dendritic Cells

  • Kim, Mi-Hyoung;Joo, Hong-Gu
    • IMMUNE NETWORK
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    • v.5 no.2
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    • pp.105-109
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    • 2005
  • Background: Dendritic cells (DCs) are the most potent APCs (antigen-presenting cells) and playa critical role in immune responses. Galectin-3 is a biological lectin with a beta-galactoside binding affinity. Recently, proteomic analysis revealed the presence of galectin-3 in the exosome of mature DCs. However, the expression and function of galectin-3 in DCs remains unclear yet. Methods: We used bone marrow-derived DCs of mouse and showed the expression of galectin-3 in DCs by using flow cytometry analysis and Western blot analysis. Results: Galectin-3 was determined as single band of 35 kDa in Western blot analysis. Flow cytometry analysis showed the major growth factor for DCs, granulocyte-macrophage colony stimulating factor (GM-CSF) and maturing agents, anti-CD40 monoclonal antibody (mAb) and lipopolysaccharide (LPS) consistently increased the intracellular expression of galectin-3 in DCs compared to medium alone. In addition, DCs treated with maturing agents did marginally express galectin-3 on their surface. Conclusion: This study suggests that galectin-3 in DCs may be regulated by critical factors for DC function.

Measurement of Bacterial (Escherichia coil) Concentration by Flow Cytometry

  • Ji, Suk;Lee, Jung-Ok;Choi, Young-Nim
    • International Journal of Oral Biology
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    • v.30 no.2
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    • pp.65-69
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    • 2005
  • Periodontitis is a multi-microbial disease and the comparison of a series of periodontopathogenic and non-periodontopathogenic bacteria in terms of microbe-host interaction may provide clues to understand the microbial etiology of the disease better. When we deal with twenty different bacterial species in a study, the first technical issue is how to measure the accurate concentration and use the same number of bacterial cells. We measured bacterial concentration by enumerating bacteria stained with SYTOX green for constant time using a flow cytometer and compared the results with those obtained by plate counting. Concentrations calculated by two different methods were very close. Therefore, flow cytometric counting allowed the rapid analysis of live/dead bacteria, offering the advantage of turbidity measurement and that of colony counting together.

성장중인 수토끼에서 혈청 IGF-I 수준과 Flow Cytometry 측정 정자형성의 변화

  • 김창근;이주형;방명걸;류재원;장유민;박민영;지달영;정영채
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.279-279
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    • 2004
  • 성장중인 수토끼에서 성성숙 전후의 혈청내 IGF-I 수준과 정자형성의 변화를 12주령부터 28주령까지 2주 간격으로 조사하였다. IGF-I 수준 측정은 non-extraction IGF-I ELISA kit(Biognostic System, Lab, USA)를 이용하였으며 정소내 정자형성은 biopsy needle(22G×3¹/₂ 72㎜×8.89㎝, Becton Dickinson, USA)로 채취한 정소조직을 fled ctyometric ploidy analysis로 DNA histogram을 4개 peak(peak Ⅰ과Ⅱ :1N, peak Ⅱ :2N, peak Ⅳ : 4N, peak Ⅲ과 Ⅳ사이 : S기)로 구분하였다. (중략)

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Estimation of nuclear DNA content of various bamboo and rattan species

  • Kumar, Prakash P.;Turner, Ian M.;Rao, A. Nagaraja;Arumuganathan, K.
    • Plant Biotechnology Reports
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    • v.5 no.4
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    • pp.317-322
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    • 2011
  • We determined the nuclear DNA content (genome size) of over 35 accessions each of bamboo and rattan species from Southeast Asia. The 2C DNA per nucleus was quantified by flow cytometry. The fluorescence of nuclei isolated from the leaves and stained with propidium iodide was measured. The genome size of the bamboo species examined was between 2.5 and 5.9 pg DNA per 2C nucleus. The genome size of the rattan species examined ranged from 1.8 to 10.5 pg DNA per 2C nucleus. This information will be useful for scientists working in diverse areas of plant biology such as biotechnology, biodiversity, genome analysis, plant breeding, physiology and molecular biology. Such data may be utilized to attempt to correlate the genome size with the ploidy status of bamboo species in cases where ploidy status has been reported.