• Journal of Aquaculture
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• v.19 no.4
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• pp.281-287
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• 2006

Intraspecific diploid androgenesis was achieved in mud loach (Misgurnus mizolepis) by the inhibition of the first mitotic division using combined thermal treatment. A combined thermal treatment (heat shock at $40.5\;^{\circ}C$ for 120 sec followed by cold treatment at $1\;^{\circ}C$ for 45 min) applied to the 1st metaphase of cell division (28 min post insemination at $25\;^{\circ}C$) successfully recovered viable androgenetic diploidy. Mean hatching success of the androgenetic diploid group was 29.6%, and the average yield out of total eggs taken was about 7% assessed at 1 week of age. However, relatively large variations in the yield of diploid androgenesis were observed among different egg batches used as cytoplasmic donors. Successful diploidization was confirmed by flow cytometric analysis, and parthenogenic reproduction in a paternal exclusive manner was verified with transgene dosage. Significant mortality was found in most androgenetic groups especially from hatch to 1 month of age, although such mortality was stabilized later.

• Microbiology and Biotechnology Letters
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• v.32 no.1
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• pp.67-71
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• 2004

Pata de Vaca (Bauhinia forficata) Is a tree which grows naturally in the rainforests and tropical parts of Peru and Brazil, as well as tropical zones of Asia, eastern Paraguay and northeastern Argentina. The active fraction (Pata-50) of the 70% ethanol extract from Pata de Vaca was sequentially fractionated by HP-20 Diaion column chromatography and C-18 column chromatography, and its characteristics were investigated. The growth of all cancer cells tested except for MCF-7 was Inhibited in a concentration-dependent manner by Pata-50. Its $IC_{50}$ values were estimated to be 40.4 $\mu\textrm{g}$/$m\ell$ on AGS, 51.3 $\mu\textrm{g}$/$m\ell$ on HT-29, 52.1$\mu\textrm{g}$/$m\ell$ on HepG2, 65.2$\mu\textrm{g}$/$m\ell$ on A549, and 77.5$\mu\textrm{g}$/$m\ell$ on HeLa cells. A flow cytometric analysis of HepG2 cells revealed induction of apoptosis, but cell cycle regulation was not affected. The HepG2 cell population of apoptosis region increased In a concentration-dependent manner by Pata-50.

• Journal of Life Science
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• v.13 no.5
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• pp.642-650
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• 2003

To develop a new approach to the treatment of neuroblastoma cells we evaluated the effect of cAMP on the Ewing's sarcoma cell line CHP-100. We observed that the proliferation-inhibitory effect of cAMP analogs was due to cell cycle arrest and induction of apoptosis, which was confirmed by observing the morphological changes and DNA fragmentation. DNA flow cytometric analysis revealed that cAMP arrested the cell cycle progression at the G1 phase, which effects were associated with inhibition of phosphorylation of retinoblastoma protein (pRB) and enhanced binding of pRB and the transcription factor E2F-1. cAMP also suppressed the cyclin-dependent kinase (Cdk) 2 and cyclin E-associated kinase activity without changes of their expressions. Furthermore, cAMP induced the levels of Cdk inhibitor $p21^{WAF1/CIP1$ expression and p21 proteins induced by cAMP were associated with Cdk2. Overall, our results identify a combined mechanism involving the inhibition of pRB phosphorylation and induction of p21 as targets for cAMP, and this may explain some of its anti-cancer effects.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.29 no.3
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• pp.134-149
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• 2016

Objectives : Gal-Geun-Tang (GT) has been described from SANGHAN in Korean traditional medicine and known to act against cold, fever, hypertension, and nasal catarrh. However, little has yet been learned about the effect of GT on immune function. In the current study, in vitro and in vivo immunomodulatory activity of GT (water extract) was investigated.Methods : Water extract of GT induced in vitro proliferation of spleen cells and significantly increased their proliferative responses during anti-CD3 activation. Using purified splenic T and B cells, it was revealed that GT has a mitogenic activity to B cells and promotes their proliferation induced by lipopolysaccharide, whereas T cell proliferation was not triggered and GT was rather inhibitory to T cell activation caused by anti-CD3 antibody. In the presence of antigen presenting cells (APC), GT addition resulted in a significant increase of IFNγ and IL-4, but not IL-2, production. However, addition of high concentration (1,000㎍/㎖) of GT led to a marked reduction in T cell cytokine production and under such condition, GT facilitated apoptosis of T cells when examined by flow cytometry with propidium iodide staining.Results : In vivo immunomdulation of GT was also investigated using a mouse model. Following keyhole limpet hemocyanin (KLH) immunization, GT (1 ㎎/day) was orally administered for 9 days. Cell numbers in thymus, spleen and peripheral blood were not altered by GT administration, indicating that such dose is not immunotoxic. Cell numbers in draining lymph nodes (LN) and ex vivo Ag-specific proliferation of LN cells were significantly elevated by GT administration. However, any preferential stimulation of T or B and CD4⁺ or CD8⁺ T cell subpopulations was not observed in a flow cytometric analysis of LN cells. This result shows that GT does not promote in vivo B cell proliferation while GT enhances Ag-specific proliferation of LN cells, unlike what was observed in vitro.Conclusions : For a further understanding of in vivo immunomodulatory activity of GT, ex vivo cytokine production of LN cells obtained from KLH-immunized mice was evaluated. Ag-specific IFNγ production was significantly higher in GT-treated mice when compared to PBS-treated control mice. In contrast, IL-4 production in GT-treated group was comparable to control group unlike to in vitro data. In addition, GT administration did not result in any significant differences in serum levels of Ig (IgM, IgG1 and IgG2a) between GT-treated and control groups. Taken together, these data strongly support that GT promotes immune response, more profoundly type 1 helper T cell (Th1) activity and GT may be applicable for treatment of intracellular parasite infection such as viral diseases.

• Korean Journal of Plant Resources
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• v.28 no.5
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• pp.656-664
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• 2015

To produce high quality watermelon, three tetraploid watermelon breeding lines (‘SA03-1’, ‘SA06-1’ and ‘SB01-1’) were developed by treatment with different chromosome doubling reagents. To identify the optimal tetraploid inductive conditions, the three watermelon breeding lines were selected by counting the number of doubled chloroplasts in guard cells. Tetraploid induction rates differed depending on the genotypes and treatment with doubling reagents. However, the highest induction rate occurred with 1.0% colchicine (82.2%). These putative tetraploid lines were re-confirmed for ploidy using flow cytometric analysis and chromosome counting. The internode length of the tetraploid breeding lines was different when the leaf size was larger in all three tetraploid lines compared to their diploids. The fruit weight of the tetraploid fruits for ‘SA03-1’ and ‘SB01-1’ was lower than for their diploid, and the rind thickness and total sugar content (°Brix) of tetraploid SB01-1 were significantly different from those of its diploid. Tetraploid lines were sterile, yielded a lower number of seeds per fruit for ‘SA03-1’ (21), ‘SA06-1’ (62), and ‘SB01-1’ (34.7), and the seeds were larger and thicker than those of their diploids. These tetraploid breeding results will be useful for breeding new seedless watermelon cultivars.

• Journal of Korean Society of Forest Science
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• v.102 no.2
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• pp.198-203
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• 2013

Herein we analyzed the nuclear DNA content and the histological characteristics of the triploid of the 'Hyunsasi' (Populus alba ${\times}$ P. glandulosa $F_1$) which were developed for biomass production and molecular breeding research. The flow cytometric analysis showed that the nuclear DNA content of the 3 triploids were 1.6 times greater than those of the diploid. In terms of histological characteristics, the cross-section area of the stem of 'Line-18' was 1.6 times larger than that of the diploid. The area of pith, and cortex and phloem of the stem of 'Line-18' was also 1.6 and 2.0 times larger than that of the diploid, respectively. Moreover, the length and area of guard cell of 'Line-18' was 1.2 times larger than that of the diploid. These results helps to understand the cytological characteristics of the triploid poplar clones, and further investigations in the growth rate and wood properties of the triploids growing in the field will determine whether the triploid poplars are good candidates for molecular breeding programs and for the improvement of industrial biomass productivity.

• Journal of Life Science
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• v.17 no.11
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• pp.1511-1516
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• 2007

Basophils and mast cells play an important role in $Fc{\varepsilon}RI-mediated$ allergic reaction as effector cells. We studied the effects of Acanthopanax chiisanensis on $Fc{\varepsilon}RI\;{\alpha}$ chain expression in human basophilic KU812F cells. Ethanol extracts from root and stem of A. chiisanensis were tested for inhibitory effects of $Fc{\varepsilon}RI\;{\alpha}$ chain expression. The cell surface $Fc{\varepsilon}RI\;{\alpha}$ chain expression was examined by flow cytometric analysis. All of the extracts of A. chiisanensis reduced the cell surface $Fc{\varepsilon}RI\;{\alpha}$ chain expression. Furthermore, A. chiisanensis extracts caused a decrease in the level of $Fc{\varepsilon}RI\;{\alpha}$ chain mRNA level and $Fc{\varepsilon}RI-mediated$ histamine release. These results suggest that root and stem extracts of A. chiisanensis play an important role in anti-allergic activity via down-regulation of $Fc{\varepsilon}RI\;{\alpha}$ chain expression and decrease in release of inflammatory mediator such as histamine.

• Journal of Life Science
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• v.17 no.11
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• pp.1492-1496
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• 2007

In vitro IgE class switching could be induced through co-culture of CD40L-expressing KU812 cells and CD40-expressing B cells in the presence of IL-4 or IL-13. It has been generated several B cell lines, which produce rice allergen (RA)-specific IgM antibody by in witγo immunization (IVI) using peripheral blood lymphocyte (PBL). In this study, induction of RA-specific IgE antibody by KU812 cells was attempted. Before co-culture, we determined the CD40 expression in RA-specific B cell lines, RA9G11 and the CD40 ligand (CD40L) expression in activated KU812 cells by treatments with phorbol myristate acetate (PMA) and ionomycin for 6 hrs. Flow cytometric analysis shown that RA9G11 and activated KU812 cells expressed high level of CD40 and CD40L, respectively. RA9G11 cells were cultured with activated KU812 cells for 12 days in the presence of IL-4 for IgE class switching. Mature $C{\varepsilon}$ mRNA level and RA-specific IgE spot forming cells (SFC) were observed in all culture condition, and especially, high level of RA-specific IgE synthesis was determined the same ratio of RA9G11 and activated KU812 cells in the presence of 50U IL-4. Therefore, induction of RA-specific IgE synthesis by activated KU812 cells can be contributed in the application for allergic therapy and prevention.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.78-88
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• 2012

Objectives: Paeonia japonica has been widely used for gynecopathy and analgesic effects in Korean Traditional Medicine. The aim of the present study is to determine the antioxidant effect of Paeonia japonica extracts(PJE) by using mouse embryonic fibroblast cells(MEF cells). Methods: We evaluated Radical Scavenging Activity of PJE by the DPPH assay. Protective effect of the PJE on the hydrogen peroxide($H_2O_2$) induced oxidative damage of MEF cells was analyzed by the MTT assay. The Morphological changes of MEF cells induced by P. japonica, $H_2O_2$ and P. japonica+$H_2O_2$ was evaluated by DAPI staining. And effect of PJE on the rate of apoptosis in MEF cells was measured using flow cytometry with Annexin V-FITC and PI double staining. Results: We observed that PJE contain significant DPPH radical scavenging activity. Cell viability of oxidative damaged cells treated with various concentrations of $H_2O_2$ was increased by treatment with PJE. Flow cytometric analysis of the cells treated with $H_2O_2$ in the absence or presence of PJE showed that the crumbled G1 peak was accumulated by the treatment with $H_2O_2$ alone, but restored by addition of PJE. Portion of cells that undergo apoptosis mediated by oxidative stress was decreased by treatment of PJE. The nuclear fragmentation occurred in the oxidative damaged MEF cells was also decreased by PJE treatment. Conclusions: Taken together, our results suggest that PJE exhibits significant antioxidant activity and functions to inhibit cell death mediated by oxidative damage induced apoptotic pathways.

• Journal of Marine Bioscience and Biotechnology
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• v.1 no.2
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• pp.63-70
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• 2006

Natural product compounds are the source of numerous therapeutic agents. The marine environment produces natural products from a variety of structural classes exhibiting activity against numerous disease targets including anticancer agents. Among these, pectenotoxin-2 (PTX-2), which was first identified as a cytotoxic entity in marine sponges, which depolymerizes actin filaments, was found to be highly effective and more potent to activate an intrinsic pathway of apoptosis in p53-deficient tumor cells compared to those with functional p53 both in vitro and in vivo. However, the anti-proliferative mechanism of the compound at non-cytotoxic concentrations has not yet been explored. In the current study, we sought to investigate anti-proliferation and apoptosis of PTX-2 against U937 human leukemic cells and its underlying molecular mechanism. Exposure of U937 cells to PTX-2 resulted in growth inhibition and induction of apoptosis in dose- and time-dependent manner as measured by MTT assay, fluorescent microscopy and flow cytometric analysis. The anti-proliferative effect of PTX-2 was associated with a marked increase in the expression of cyclin-dependent kinase p21 (WAF1/CIP1) mRNA which was tumor suppressor p53-independent. The increase in apoptosis was connected with a time-dependent down-regulation of anti-apoptotic Bcl-XL and inhibitor of apoptosis proteins (IAPs) family such as XIAP and cIAP-2. Though additional studies are needed, these findings suggested that PTX-2-induced inhibition of U937 cells was associated with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of PTX-2.