• Title/Summary/Keyword: flow cytometry

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Three Predictive Tests Using Mice for the Identification of Contact Sensitizer

  • Jung-Hyun Shin;Min
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.22 no.2
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    • pp.201-210
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    • 1996
  • Predictive tests for the identification of contact sensitizing chemicals have been developed. We measured the sensitization potential with three predictive tests, the in vitro and the in vivo Local Lymph Node Assay(LLNA), ELISA to detect interferon-gamma(IFN-${\gamma}$) from supernatant and flow cytometry to detect change of cell surface proteins, using draining lymph nodes of mice. BALB/c mice were exposed to various chemicals or vehicles on the ears daily for 3 consecutive days in all experiments. With some exceptions of propyl paraben, neomycin sulfate, the in vivo LLNA was able to detect the sensitizing capacity of test chemicals and was more sensitive than the in vitro LLNA for chemicals used in the present study. In another experiment, contact sensitivity was assessed by the ELISA to detect IFN-Υ from the supernatants of the cultured LNCs after sensitization with chemicals. There was a good correlation between the LLNA and the IFN-Υ production for test chemicals. We also examined the change of cell surface proteins on LNCs after sensitization by flow cytometry for some cell adhesion molecules(ICAM-1, E-cadherine, B7 molecule), T cell markers(CD3, CD4, CD8, T$\alpha$$\beta$,T${\gamma}$$\delta$) and B cell markers(LR1, CD45R, I-Ad). The number of ICAM-1 positive cells and B cells in LNCs were increased after sensitization with DNCB, TNCB, isoeugenol and 25%, 50% cinnamic aldehyde compared with that of vehicle as a control. In conclusion, the in vivo LLNA could provide more sensitive screening test for moderate to strong sensitizers and some weak sensitizers including cosmetic raw materials than the in vitro LLNA. The production of IFN-Υ by allergen-activated LNCs might be a values indicators without radioisotopes for the identification of contact allergens. Detection of allergens by testing the increase of ICAM-1 positive cells and B cells in LNCs by flow cytometry might be used as a test method to detect allergens.

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IRRADIATION EFFECT ON THE APOPTOSIS INDUCTION IN THE HUMAN CANCER CELL LINES AND THE GINGIVAL FIBROBLAST (인체의 암세포주와 치은섬유모세포주에서 방사선조사가 apoptosis 유발에 미치는 영향에 관한 연구)

  • Park Moo-Soon;Lee Sam-Sun;Choi Soon-Chul;Park Tae-Won;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.28 no.1
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    • pp.59-71
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    • 1998
  • The radiation-induced apoptosis was studied for two human cancer cell lines (KB cells, RPMI 2650 cells) and the human gingival fibroblast cell line (HGF-1 cells). The single irradiation of 2, 10, 20Gy was done with 241.5 cGylmin dose rate using the /sup 137/Cs MK cell irradiator. The cells were stained with propidium iodide and examined under the fluoro-microscope and assayed with the flow cytometry a day after irradiation. Also, the LDH assay was done to determine the amount of necrotic cells. The obtained results were as follows: 1. On the fluoro-microscope, many fragmented nuclei were detected in the KB, RPMI 2650, and HGF-1 cells after irradiation. 2. On the DNA content histogram obtained from the flow cytometry, the percentages of the pre-Gl peak of the control and 2, 10 and 20Gy irradiation group were 4.5, 55.0, 52.3, and 66.6% on KB cells, 2.7, 3.3, 31.8, and 32.6% on RPMI 2650 cells and 2.8, 21.8, 30.4, and 40.2% on HGF-1 cells respectively. 3. The number of Gl-stage cells was abruptly decreased after 2Gy irradiation on KB cells and 10Gy irradiation on RPMI 2650 cells, But there was a slight decrease without regard to irradiation dose on HGF-1 cells. 4. There was no significantly different absorbance in extracellular LDH assay along the experimental cell lines.

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Cytogenetic Analysis of the Triploid Pacific Abalone, Haliotis discus hannai (북방전복, Haliotis discus hannai 3배체의 세포유전학적 연구)

  • Jee, Young-Ju;Chang, Young-Jin
    • The Korean Journal of Malacology
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    • v.28 no.1
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    • pp.37-43
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    • 2012
  • In this study, we invesgated a cytogenetic analysis of the Pacific triploid abalone, Haliotis discus hannai induced by low temperature treatment. We got a lot of mitotic metaphase chromosome spreads from the triploid and diploid Pacific abalones' hatched larvae (trochophores). The chromosome number of diploid abalone was 2n = 36 and that of triploid abalone was 3n = 54, so the chromosome number of triploid abalone was 1.5 times higher than that of diploid abalone. We developed a modified flow cytometric method for Pacific abalone from the existing methods. We uesd 51 months aged triploid and diploid Pacific abalones' hemolymph to get the DNA contents by flow cytometry. The DNA content of diploid abalone was 1.743 pg/cell and the DNA content of triploid abalone was 1.49 times higher than that of diploid one. It proved that triploid abalone was consisted with two sets of maternal diploid and one set of paternal genome.

Studies on the Effect of Corilagin Isolated from Euphorbia helioscopia on Collagen-Induced Arthritis (I);Analysis of Fluorescence Flow Cytometry from Collagen II Induced Arthritis Mice Model (택칠에서 분리한 Corilagin이 Collagen 유발 관절염에 미치는 영향 (I);Corilagin을 투여한 류마티스 관절염 유발 생쥐의 형광유세포 분석)

  • Shin, Sam-Kee;Jang, Jun-Pok;Doh, Eun-Soo
    • Korean Journal of Plant Resources
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    • v.21 no.4
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    • pp.329-335
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    • 2008
  • This study was intended to research into the possibility of corilagin (CRN) isolated from Euphorbia helioscopia as rheumatoid arthritis drug. For the study, CRN was medicated to the abdominal cavity of collagen induced arthritis (CIA) mice that was a animal model for rheumatoid arthritis and its effects on incidence and arthritis index (AI) were studied. The results were as folllows; Medicating corilagin that were isolated from Euphorbia heliscopia to CIA mice resulted in inhibiting an incidence of arthritis and reducing in arthritis index. It was found that inflammatory cells were remarkably decreased and joint cavity was well secured. Also it was displayed that the formation of pannus was not observed and a cartilage was preserved well by cell histological research. Performing fluorescence flow cytometry of CIA mice resulted in reducing in inflammatory cells infiltrated each tissue.

Protoclonal variation in cabbage (Brassica oleracea ssp. capitata) (양배추 protoclone의 변이)

  • 이연희;조현석;김호일;나종현;이인원
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.175-181
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    • 1997
  • Plants were regenerated from hypocotyl protoplast culture of cabbage (F$_1$hybrid Green Challenger) and were transplanted on fields. The ploidy level of regenerated and seed-grown plants was measured by flow cytometry. In total 125 regenerated plant, diploid (2n), tetraploid(4n), and mixoploid (2n+4n) were 72.8%, 25.6%, and 1.6%, respectively. Most of the regenerated plants had normal phenotype, whereas several plane showed abnormal phenotype such as heavy leaf incision, savoy, and wave. The regenerated plants with abnormal phenotype and different ploidy level were analysed by isozyme and RAPD, but no significant difference was found.

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Non-specific Defensive Factors of the Pacific Oyster Crassostrea gigas against Infection with Marteilioides chungmuensis: A Flow-Cytometric Study

  • Choi, Hee-Jung;Hwang, Jee-Youn;Choi, Dong-Lim;Huh, Min-Do;Hur, Young-Baek;Lee, Nam-Sil;Seo, Jung-Soo;Kwon, Mun-Gyeong;Choi, Hye-Sung;Park, Myoung-Ae
    • The Korean Journal of Parasitology
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    • v.49 no.3
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    • pp.229-234
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    • 2011
  • In order to assess changes in the activity of immunecompetency present in Crassostrea gigas infected with Marteilioides chungmuensis (Protozoa), the total hemocyte counts (THC), hemocyte populations, hemocyte viability, and phagocytosis rate were measured in oysters using flow cytometry. THC were increased significantly in oysters infected with M. chungmuensis relative to the healthy appearing oysters (HAO) (P<0.05). Among the total hemocyte composition, granulocyte levels were significantly increased in infected oysters as compared with HAO (P<0.05). In addition, the hyalinocyte was reduced significantly (P<0.05). The hemocyte viability did not differ between infected oysters and HAO. However, the phagocytosis rate was significantly higher in infected oysters relative to HAO (P<0.05). The measurement of alterations in the activity of immunecompetency in oysters, which was conducted via flow cytometry in this study, might be a useful biomarker of the defense system for evaluating the effects of ovarian parasites of C. gigas.

Expression of Chemokines and Chemokine Receptors in Brain Tumor Tissue Derived Cells

  • Razmkhah, Mahboobeh;Arabpour, Fahimeh;Taghipour, Mousa;Mehrafshan, Ali;Chenari, Nooshafarin;Ghaderi, Abbas
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.17
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    • pp.7201-7205
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    • 2014
  • Chemokine and chemokine receptor expression by tumor cells contributes to tumor growth and angiogenesis and thus these factors may be considered as tumor markers. Here we aimed to characterize cells directly extracted from glioma, meningioma, and secondary brain tumors as well as non-tumoral cells in vitro. Cells were isolated from brain tissues using 0.2% collagenase and characterized by flow cytometry. Expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, MCP-1 and IP-10 was defined using flow cytometry and qRT-PCR methods. Brain tissue isolated cells were observed as spindle-shaped cell populations. No significant differences were observed for expression of SDF-1, CXCR4, CXCR7, RANTES, CCR5, and IP-10 transcripts. However, the expression of CXCR4 was approximately 13-fold and 110-fold higher than its counterpart, CXCR7, in meningioma and glioma cells, respectively. CXCR7 was not detectable in secondary tumors but CXCR4 was expressed. In non tumoral cells, CXCR7 had 1.3-fold higher mRNA expression than CXCR4. Flow cytometry analyses of RANTES, MCP-1, IP-10, CCR5 and CXCR4 expression showed no significant difference between low and high grade gliomas. Differential expression of CXCR4 and CXCR7 in brain tumors derived cells compared to non-tumoral samples may have crucial impacts on therapeutic interventions targeting the SDF-1/CXCR4/CXCR7 axis.

Application of Multiparametric Flow Cytometry (FCM) to Enumerate the Diagnosis of Pseudomonas aeruginosa and Escherichia coli

  • Hwang, Myoung-Goo;Oh, Jung-Woo;Katayama, Hiroyuki;Ohgaki, Shinichiro;Cho, Jin-Kyu
    • Environmental Engineering Research
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    • v.17 no.1
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    • pp.35-39
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    • 2012
  • In this study, multiparametric flow cytometry (FCM) was installed to enumerate the diagnosis of Pseudomonas aeruginosa ATCC 10145 and Escherichia coli K12 (IFO 3301). The nucleic acids (DNA/RNA) were double stained by a LIVE/DEAD bacLight viability kit, involving green SYTO 9 and red propidium iodide (PI), based on the permeability of two chemicals according to the integrity of plasma membrane. As the results showed, the gate for dead bacteria was defined as the range of $0.2{\times}10^0$ to $6.0{\times}10^1$ photo multiplier tube (PMT) 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $2.0{\times}10^2$ PMT 4 fluorescence (Y-axis), and the gate for live bacteria was defined as the range of $6.0{\times}10^0$ to $6.0{\times}10^2$ PMT 2 fluorescence (X-axis) and $2.0{\times}10^0$ to $4.0{\times}10^2$ PMT 4 fluorescence (Y-axis). In the comparison of the number of the tested bacteria detected by FCM (viability assessment) and plate culture (cultivability assessment), the number of bacteria detected by FCM well represented the number of bacteria that was detected by the colony forming unit (CFU) counting method when bacteria were exposed to isopropyl alcohol and silver/copper cations. Consequently, it is concluded that the application of FCM to monitor the functional effect of disinfectants on the physiological status of target bacteria can offer more rapid and reliable data than the plate culture colony counting method.

Expression and Characterization of Protein Latcripin-3, an Antioxidant and Antitumor Molecule from Lentinula edodes C91-3

  • Ann, Xiao-Hua;Lun, Yong-Zhi;Zhang, Wei;Liu, Ben;Li, Xing-Yun;Zhong, Min-Tao;Wang, Xiao-Li;Cao, Jing;Ning, An-Hong;Huang, Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.5055-5061
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    • 2014
  • In this study, an anti-oxidant and anti-tumor protein Latcripin-3 of Lentinula edodes C91-3 was expressed in Escherichia coli. for the first time. According to the cDNA library, the full-length gene of Latcripin-3 was cloned by the methods of 3'-full rapid amplification of cDNA Ends (RACE) and 5'-full RACE. The structural domain gene of Latcripin-3 was inserted into the pET32 a(+). The functional protein of Latcripin-3 was expressed in Rosetta-gami (DE3) E. coli, evaluated by Western blotting and mass spectrometry. DPPH testing showed that the protein Latcripin-3 can scavenge free radicals remarkably well. The activity of functional protein Latcripin-3 on A549 cells was studied with flow cytometry and the MTT method. The MTT assay results showed that there was a decreases in cell viability in a dose-dependent and time-dependent manner in protein Latcripin-3 treated groups. Flow cytometry demonstrated that Latcripin-3 can induce apoptosis and block S phase dramatically in human A549 lung cancer cells as compared to the control group. At the same time, the cell ultrastructure observed by transmission electron microscopy supported the results of flow cytometry. This research offers new insights and advantages for identifying anti-oxidant and anti-tumor proteins.

Exploring natural hybridizations among Asplenium ruprechtii and related taxa in Korea

  • LEE, Chang Shook;YEAU, Sung Hee;CHUNG, Kyong-Sook
    • Korean Journal of Plant Taxonomy
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    • v.49 no.2
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    • pp.127-139
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    • 2019
  • The purported four hybrid origins of Asplenium in Korea were tested based on morphological, cytological and DNA sequence data. Asplenium castaneo-viride, A. ${\times}$ uiryeongse, A. ${\times}$ montanus, and A. ${\times}$ kitazawae share several morphological characteristics with the Asian walking fern A. ruprechtii and related taxa as parents and show a sympatric distribution with the putative parents, raising the possibility of hybrid origins: A. castaneo-viride (A. ruprechtii and A. incisum), A. ${\times}$ uiryeongse (A. ruprechtii and A. pekinense), A. ${\times}$ montanus (A. ruprechtii, A. trichomanes, and A. incisum), and A. ${\times}$ kitazawae (A. ruprechtii and A. sarelii). We investigated flow cytometry and chloroplast DNA sequence data (rbcL, rps4-trnS, and rps4-trnS intergenic spacer) to clarify the hybridization and origin of each hybrid. In the flow cytometry analyses, A. ruprechtii shows diploid (2x) only, whereas A. castaneo-viride (3x, 4x), A. ${\times}$ uiryeongse (3x), A. ${\times}$ montanus (3x, 4x), and A. ${\times}$ kitazawae (2x, 4x) exhibit polyploidy, suggesting hybrid events along speciation. The rbcL and rps4-trnS and rps4-trnS intergenic spacer data suggest that A. ruprechtii is one the maternal ancestors of all four hybrids. In addition, the rps4-trnS and rps4-trnS intergenic spacer data indicate that A. incisum is also the maternal ancestor of A. ${\times}$ kitazawae and A. ${\times}$ montanus, proposing multiple hybridization events for these two hybrids. In A. ${\times}$ montanus, morphological features such as the leaf forms and sympatric distributions of the species also support the multimaternal hypothesis, but the morphological features of A. ${\times}$ kitazawae must be examined with consideration of hybrid events. To clarify the complex hybrid evolutionary lineages of the four Asplenium hybrids, further research with taxon sampling and molecular markers should be conducted.