• KOREAN JOURNAL OF CROP SCIENCE
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• 제58권4호
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• pp.331-335
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• 2013

Buckwheat sprout is used as vegetable, and also flour for making noodles, and so on. Currently, information about tissue culture in buckwheat is limited and restricted to micro-propagation. We carried out somatic embryogenesis and plant regeneration using hypocotyl segments as explant of the cultivated buckwheat species, Fagopyrum esculentum which differs from existing studies in the growth regulator combinations used. Maximum callus regeneration was induced on MS medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) $2.0mg{\cdot}L^{-1}$, benzyladenine (BA) $1.0mg{\cdot}L^{-1}$ and 3% sucrose. Friable callus was transferred to solidified MS media containing BA ($1.0mg{\cdot}L^{-1}$) with various concentrations of 2,4-dichlorophenoxyacetic acid for the induction of embryogenesis. The optimum concentrations of growth regulators (for regeneration of plantlet) were indole-3-acetic acid ($2.0mg{\cdot}L^{-1}$), Kinetin ($1.0mg{\cdot}L^{-1}$), BA ($1.0mg{\cdot}L^{-1}$). Only 2,4-D did not show any significant effect on callus induction or embryogenesis. Regeneration of embryonic callus varied from 5% to 20%. Whole plants were obtained at high frequencies when the embryogenic calli with somatic embryos and organized shoot primordia were transferred to MS media with 3% sucrose. The main objective of this research was to develop an efficient protocol for plant regeneration for common buckwheat, and to apply in future for genetic transformation.

• Journal of Society of Preventive Korean Medicine
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• 제16권2호
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• pp.95-111
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• 2012

Objective : The water extract of Danguiyonghoihwan (DGYHW) has been traditionally used in treatment of tinnitus in Oriental Medicine. However, little is known about the mechanism by which DGYHW rescues auditory neuronal cells from injury damages. Therefore, in this study I effort to elucidate the mechanism of the cytoprotective effect of the DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Methods : I determined the elevated cell viability by DGYHW extract on glutamate-induced auditory sensorineuronal cell death. Glutamate induced neuronal damage in oranotypic explant culture also, glutamate decreased cell viability on VOT-33 cells but pretreatment with DGYHW inhibited cell death. Results : One of the main mediator of glutamate-induced cytotoxicity was known to generation of reactive oxygen species (ROS). Pretreatment with DGYHW inhibited this ROS generation from glutamated-stimulated VOT-33 cells. Also, I identified that the ROS-induced DCF-DA green fluorescence is reduced by DGYHW pretreatment. The critical markers of apoptotic cell death were cleavages of procaspase-3 protease protein. So I checked the expression level and cleavage of procaspase-3 protease protein. Glutamate-treated VOT-33 cells were shown to have cleavage of procaspase-3 protease proteins and following reduction of expression of these proteins. But I found that pre-treatment with DGYHW protects glutamate-induced changes of biochemical marker protein, caspase-3. Conclusion : These findings indicated that DGYHW may prevent cell death from glutamate induced VOT-33 cell death by inhibiting the ROS generation and modulation of protein expressions in procaspase-3, catalase and Bcl-2.

• Korean Journal of Plant Resources
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• 제10권1호
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• pp.94-99
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• 1997

When the leaf and stem tissues of Rehmannia glutinosa were cultured on MS medium with plant growth regulators, shoots were regenerated on MS medium with TDZ(0.01 to $2.0{\mu}M$), and root initiation was better on MS medium treated with NAA(0.01 to 2.0mg/$\iota$). On $B_5$ medium, shoot regeneration was better on medium with TDZ than those with Quincrac, NAA, and 2.4-D. The addition of Quincrac, NAA, and 2.4-D inhibited shoot regeneration on MS medium, but promoted shoot regeneration on $B_5$ medium. Shoot regeneration and growth was better on MS medium with the combination treatments of 2.4-D 0.01mg/$\iota$ and TDZ 0.01, 0.1 and $2{\mu}M$ compared with other treatments. Also shoot regeneration and growth on B_5 medium showed the similar results to that on MS medium.

• Korean Journal of Plant Resources
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• 제16권1호
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• pp.34-39
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• 2003

Corm apical meristem cultures of thirteen glasshouse freesia cultivars were tested to investigate the possibility of micropropagation using MS basal medium supplemented with 2,4-D, NAA(0.1, 0.5, 1.0mg/L, respectively) and BA (0.5∼2.0mg/L). The majority of the tested cultivars could be induced callus and shoot buds in all culture condition. The combinations of NAA and BA appeared superior to that of 2,4-D and BA depending on cultivars for callus induction and shoot formation. Among the cultivars, 'Golden Yellow' showed the highest regeneration capacity on MS media with 0.5mg/L NAA and 1.0 mg/L BA. The highest percentage of regeneration and the greatest number of shoot from calli were obtained through successive subculture on MS medium supplimented with 0.5mg/L BA. In that condition, more than 60 ％ shoot regeneration and average of 25.1 shoots per explant was achieved. Transformed shoots on half-strength MS medium without plant growth regulators rooted easily.

• KOREAN JOURNAL OF CROP SCIENCE
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• 제49권3호
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• pp.232-236
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• 2004

In order to optimize tissue culture conditions for genetic transformation of orchardgrass (Dactylis glomerata L.), the effects of culture medium supplements on tissue culture responses were investigated with mature seeds of a cultivar, 'Roughrider', as explant tissues. The optimal concentration of 2,4-D for the induction of embryogenic callus from mature seeds was 3 mg/L. Plant regeneration frequency was 36.3% when embryogenic calli were cultured on the regeneration medium supplemented with 1mg/L 2,4-D and 3mg/L BA. Addition of 1 g/L casein hydrolysate and 300 mg/L L-proline improved frequencies of embryogenic callus induction and plant regeneration up to 57.3 and 60.7%, respectively. Supplementation of the media with 10 mga $\textrm{AgNO}_3$ and 40 mg/L cysteine enhanced frequencies of callus induction and plant regeneration. Efficient regeneration system established in this study will be useful for molecular breeding of orchardgrass through genetic transformation.

• Korean Journal of Medicinal Crop Science
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• 제4권2호
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• pp.132-138
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• 1996

This experiment was carried out to establish a year-round production system of pathogen-free stock through micropropagation in Fritillaria thunbergii as medicinal bulbous plant. The effect of different types of culture method and plant growth regulators, activated charcoal and mannitol on bulblet formation and subsequent growth were investigated. The MS solid medium containing 1. 0 mg/L kinetin and 0. 3 mg/L NAA was effective on bulblet formation and propagation rate compared to liquid and suspension culture. Addition of activated charcoal at 0. 01% to 0. 1% in the medium promoted bulbing of cultured bulblets and shoot formation. Addition of 1% to 2% mainnitol in MS medium was effective on the formation of bulblet and subsequent growth of bulblets compared to control. In addition of inhibitors, $10{\sim}100\;mg/L$ B-9 and Chloromequat had effective to stimulate bulblet growth but their effects were not so much as mannitol treatment. ABA treatment had detrimental effect on survival rate of explant and bulblet formation.

• Journal of Periodontal and Implant Science
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• 제24권2호
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• pp.238-254
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• 1994

Cyclosporine A is an immunosuppressant commonly used for patients receiving organ transplants. Gingival overgrowth is an adverse side-effect seen in about 8-26% of patients taking cyclosporine A which have been shown to increase the DNA synthesis of gingival fibroblast at the concentration of $10^{-9}g/ml$ in vitro. Glycyrrhetinic acid is the active pharmacological ingredients of licorice which exerts steroid-like action and anti-viral activity. Oleanolic acid, which were isolated from Glechoma hederacea, has been shown to act as inhibitors of tumor promotion in vivo and to be less cytotoxic retinoic acid. This study has been performed to evaluate the effects of glycyrrhetinic acid and oleanolic acid on cyclosporine A induced cell activity in vitro. Human gingival fibroblasts were isolated from explant cultures of healthy gingiva of orthodontic patients. Gingival fibroblasts were trypsinized and transferred to the walls of microtest plates. Fibroblasts were cultured in growth medium added $10^{-9}g/ml$ cyclosporineA and $50{\mu}l/ml$ lipopolysaccharides. Cells between the 4th and 6th transfer in culture were used for this study. The morphology of gingival fibroblst were examined by inverted microscope. The effects of cyclosporine A on the time course of DNA sythesis by human gingival fibroblasts were assessed by $[^3H]-thymidine$ uptake assays. Cyclosporine A was found to stimulate DNA synthesis of human gingival fibroblast at a concentration of $10^{-9}g/ml$. In the presence of lipopolysaccharide derived from Fusobacterium nucleatum, addition of cyclosporine A results in reversal of inhibition at the concentration which normally inhibits gingival fibroblast proliferation. The cell acitivities in the presence of glycyrrhetinic acid and oleanolic acid were decreased, and increased cell acitivities by cyclosporine A were decreased by glycyrrhetinic acid and oleanolic acid at the concentration of $200{\mu}g/ml$. These results suggested that the increased cell activities by cyclosporine A modulated by glycyrrhetinic acid and oleanolic acid.

• Asian-Australasian Journal of Animal Sciences
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• 제24권7호
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• pp.919-928
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• 2011

The pig exhibits true epitheliochorial placentation, where the fetal membrane maintains attachment throughout pregnancy but does not invade into the maternal uterine endometrium. Accordingly, the expression and function of cell adhesion molecules are very important for embryo implantation and the establishment of pregnancy. In our recent microarray analysis, we found that activated leukocyte cell adhesion molecule (ALCAM) was expressed in the uterine endometrium during pregnancy in pigs. To better understand the roles of ALCAM in the establishment and maintenance of pregnancy, we examined ALCAM expression in the uterine endometrium during the estrous cycle and pregnancy in pigs. Real-time RT-PCR analysis showed that ALCAM was differentially expressed in the uterine endometrium during the estrous cycle and pregnancy, with the highest levels on D12 of pregnancy. ALCAM mRNA was localized to the luminal and glandular epithelial cells and to the trophectoderm of conceptuses during early pregnancy. The steroid hormones estrogen and progesterone had no effect on ALCAM expression in an endometrial explant culture study. Further, we found that ALCAM expression in the uterine endometrium from gilts with somatic cell nuclear transfer-derived embryos was not different from that in gilts with embryos from natural mating. ALCAM was expressed in a pregnancy stage- and cell type-specific manner in the uterine endometrium and conceptuses during pregnancy. These findings suggest that ALCAM may play a role in the establishment of pregnancy. Further analysis of ALCAM will provide insight into the implantation process and establishment of pregnancy in pigs.

• Journal of Korean Society of Forest Science
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• 제90권2호
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• pp.184-189
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• 2001

Multiple shoots were induced from in vitro shoot originated from winter bud of P. yedoensis from Jeju. Most explants grow in similar type among the five different media but affected by supplement of sucrose regardless of media. For mass-propagation various concentrations of BAP or $GA_3$ were treated in the medium respectively. BAP was very effective to produce multiple shoots and 3.5~9.5 shoots were formed on the explant. The shoots induced on the high levels of BAP have short internodes. No shoots were induced on the treatment of $GA_3$ but roots were induced on it. When $GA_3$ was supplemented with the medium containing BAP, multiple shoots were produced from the explants. The medium(WPM) containing with $0.5mg/{\ell}$ BAP and $4.0mg/{\ell}$ $GA_3$ was most effective to produce multiple shoots. When the explants were cultured for 8 weeks, 39.5 shoots were developed in average.

• Journal of Crop Science and Biotechnology
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• 제11권2호
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• pp.119-126
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• 2008

Since somatic embryogenesis combined with ethylmethane sulfonate(EMS) treatments is the most efficient technique for mutagenesis, the embryogenic capacity of four soybean cultivars was evaluated at different EMS concentrations, treatment times, and preculture durations. Two to 4 mm long immature cotyledons were placed in induction medium after EMS treatment, and the numbers of somatic embryos formed per explant were counted four weeks after culture initiation. We observed genotypic differences in the efficiency of somatic embryogenesis from immature embryos among four cultivars treated with different concentrations of EMS for six hours. Cultivars, Sinpaldalkong 2 and Jack, displayed highly efficient somatic embryogenesis regardless of EMS concentration, whereas very low efficiency or no survival was observed in Jinju 1 and Iksannamulkong cultivars. Preculture duration did not influence the efficiency of somatic embryogenesis. Because Sinpaldalkong 2 exhibited the best somatic embryogenesis, much higher concentrations of EMS were used to test somatic embryo formation under different periods of time in this cultivar. Three and six hour treatments with both 1 and 2 mM EMS yielded higher embryo formation than longer periods of time. Increasing the time with embryos in 2 mM EMS caused a reduction in somatic embryogenesis in Sinpaldalkong 2, but many chlorophyll-deficient soybean variants were identified in the $M_1R_0$ and $M_2R_1$ generations. In addition to Jack, Sinpaldalkong 2 is a good genotype for plant regeneration from EMS-treated immature embryo cultures.