• Journal of Periodontal and Implant Science
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• v.26 no.1
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• pp.1-26
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• 1996

This study was performed to estimate the effects of cultured bone cell inoculated on porous type hydroxyaptite for the regeneration of the artificial alveolar bone defect. In this experiment 3 beagle dogs were used, and each of them were divided into right and left mandible. Every surgical intervention were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). To reduce the gingival bleeding during surgery, operative site was injected with Lidocaine hydrochloride(l:80,000 Epinephrine) as local anesthesia. After surgery experimental animal were feeded with soft dietl Mighty dog, Frisies Co., U.S.A.) for 1 weeks to avoid irritaion to soft tissue by food. 2 months before surgery both side of mandibular 1st premolar were extracted and bone chips from mandibular body were obtained from all animals. Bone cells were cultured from bone chips obtained from mandible with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Porous type hydroxyapatite were immerse into the high concentrated cell suspension solution, and put 4 hours for attachin the cells on the surface of hydroxyapatite. Graft material were inserted on the artificial bone defect after 3 days of culture. Before insertion of cellinoculated graft material, scanning electronic microscopic observation were performed to confirm the attachment and spreading of cell on the hydroxyapatite surface. 3 artificial bone defects were made with bone trephine drill on the both side of mandible of the experimental animal. First defect was designed without insertion of graft material as negative control, second was filled with porous replamineform hydroxyapatite inoculated with cultured bone marrow cells as expermiental site, and third was filled with graft materials only as positive control. The size of every artificial bone defect was 3mm in diameter and 3mm in depth. After the every surgical intervention of animals, oral hygiene program were performed with 1.0% chlorhexidine digluconate. All of the animals were sacrificed at 2, 4, 6 weeks after surgery. For obtaining histological section, tissus were fixed in 10% Buffered formalin and decalcified with Planko - Rycho Solution for 72hr. Tissue embeding was performed in paraffin and cut parallel to the surface of mandibular body. Section in 8um thickness of tissue was done and stained with Hematoxylin - Eosin. All the specimens were observed under the light microscopy. The following results were obtained : 1. In the case of control site which has no graft material, less inflammatory cell infiltration and rapid new bone forming tendency were revealed compared with experimental groups. But bone surface were observed depression pattern on defect area because of soft tissue invasion into the artificial bone defect during the experimental period. 2. In the porous hydroxyapatite only group, inflammatory cell infiltration was prominet and dense connective tissue were encapsulated around grafted materials. osteoblastic activity in the early stage after surgery was low to compared with grafted with bone cells. 3. In the case of porous hydroxyapatite inoculated with bone cell, less inflammatory cell infiltration and rapid new bone formation activity was revealed than hydroxyapatite only group. Active new bone formation were observed in the early stage of control group. 4. The origin of new bone forming was revealed not from the center of defected area but from the surface of preexisting bony wall on every specimen. 5. In this experiment, osteoclastic cell was not found around grafted materials, and fibrovascular invasion into regions with no noticeable foreign body reaction. Conclusively, the cultured bone cell inoculated onto the porous hydroxyapatite may have an important role of regeneration of artificial bone defects of alveolar bone.

• Korean Journal of Environmental Agriculture
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• v.15 no.2
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• pp.167-178
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• 1996

In order to identify PCP glucose conjugates transformed from PCP in soybean and rice cell suspension cultures, the purified metabolites were acetylated, purified twice by HPLC using a normal and a reversed phase column, and then subjected to fast atom bombardment(FAB) mass spectrometric analysis. As were the conjugates, their acetylated derivatives of the glucose conjugates formed at the early stage(48 hr) of metabolism were separated by HPLC into three fractions. FABMS analysis of each fraction revealed that, at least in two fractions, the locations of the spectral peaks were practically coincident with those deducible from the structures of pentachlorophenyl and tetrachlorophenyl ${\beta}-D-glucopyranosides$. Based on information obtained from mass spectral and chromatographic analysis of not only the water-soluble metabolites but also aglycones and glycone, it is concluded that PCP is primarily metabolized to glucose conjugates, which account for more than 50% recovery of the PCP-conveyed radioactivity from the water soluble metabolites : The conjugates are mainly made up of pentachlorophenyl ${\beta}-D-glucopyranoside$, tetrachlorophenyl ${\beta}-D-glucopyranosides$( probably 2 or more isomers), and 2-hydroxy-3,4,5,6-tetrachlorophenyl ${\beta}-D-glucopyranoside$.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.335-345
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• 2020

Tomato grey mould has been one of the destructive fungal diseases during tomato production. Ten mM of menadione sodium bisulfite (MSB) was applied to tomato plants for eco-friendly control of the grey mould. MSB-reduced tomato grey mould in the 3rd true leaves was prolonged at least 7 days prior to the fungal inoculation of two inoculum densities (2 × 10⁴ and 2 × 10⁵ conidia/ml) of Botrytis cinerea. Protection efficacy was significantly higher in the leaves inoculated with the lower disease pressure of conidial suspension compared to the higher one. MSB-pretreatment was not effective to arrest oxalic acid-triggered necrosis on tomato leaves. Plant cell death and hydrogen peroxide accumulation were restricted in necrotic lesions of the B. cinereainoculated leaves by the MSB-pretreatment. Decreased conidia number and germ-tube elongation of B. cinerea were found at 10 h, and mycelial growth was also impeded at 24 h on the MSB-pretreated leaves. MSB-mediated disease suppressions were found in cotyledons and different positions (1st to 5th) of true leaves inoculated with the lower conidial suspension, but only 1st to 3rd true leaves showed decreases in lesion sizes by the higher inoculum density. Increasing MSB-pretreatment times more efficiently decreased the lesion size by the higher disease pressure. MSB led to inducible expressions of defence-related genes SlPR1a, SlPR1b, SlPIN2, SlACO1, SlChi3, and SlChi9 in tomato leaves prior to B. cinerea infection. These results suggest that MSB pretreatment can be a promising alternative to chemical fungicides for environment-friendly management of tomato grey mould.

• Korean Journal of Plant Resources
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• v.30 no.5
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• pp.542-548
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• 2017

Two different races of Botryris cinerea were selected by the response of plant leaves to the pathogen infection. Based on lesion size of the pathogen on the leaves, turnip showed susceptible response to 'Grape-01' race, and resistant to 'Orange' race. Turnip leaves infected with resistant pathogen race, "Orange", showed significantly higher content of indole-3-ylmethyl glucosinolate (I3M) than those infected with susceptible race, 'Grape-01'. Contents of I3M in the leaves with resistant 'Orange' race was 2.5 times as high as that in uninfected leaves, whereas I3M in the leaves infected with susceptible 'Grape-01' race showed lower content than in untreated leaves. Growth of turnip suspension cells was significantly inhibited by the treatment of MeOH extract or water extract of 'Orange' race as compared with the treatment of susceptible race, 'Grape-01'. Treatment of MeOH or water extract from 'Orange' race to turnip suspension cells, strongly inhibited cell viability up to 22.7% or 16.5%, respectively. However, plant cells treated with MeOH or water extract from resistant race, 'Orange' showed higher I3M content than that from susceptible race, 'Grape-01'. These results suggest that accumulation and degradation of I3M glucosinolate in turnip cells closely related to the resistance and susceptibility of turnip cells to Botrytis cinerea.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.5
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• pp.660-664
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• 2011

We investigated the improvement of the antioxidative activity of black garlic with enzymatic treatment and lactic acid bacteria cultivation conditions. Celluclast, a commercially-available polysaccharide hydrolyase, was selected to obtain high total polyphenol content in a black garlic suspension. A lactic acid bacterial strain showing fast growth and high acid production in a black garlic suspension was isolated from Kimchi. This strain was identified as Lactobacillus pentosus 310-7. Enzymatically hydrolyzed black garlic was fermented using the L. pentosus 310-7 strain at $30^{\circ}C$ for 15 hr. The pH and titratable acidity achieved were 4.24 and 0.35%, respectively, after 15 hr fermentation. The viable cell population of L. pentosus 310-7 slowly increased to 7.54 log CFU/g. The polyphenolic compound content, known antioxidants, in black garlic was enhanced with Celluclast treatment and L. pentosus 310-7 cultivation. Total polyphenolic compounds were increased to approximately 60% of the initial concentration, and electron donating ability was also improved, from 39.8 to 65.9%.

• Korean Journal of Plant Resources
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• v.24 no.4
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• pp.461-471
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• 2011

This study was carried out to establish an in vitro culture method of callus having a high antioxidant activity from Ginkgo biloba L. Leaf explants were cultured on Murashige and Skoog's medium supplemented with various growth regulators. The explants were incubated in the dark or 3,000 lux cool-white light. Methanol extracts from incubated callus were evaluated for scavenging activity of the free radicals using DPPH. The best callus growth rate was achieved in MS medium combined with 10 ${\mu}M$ NAA and 5 ${\mu}M$ kinetin in the light condition. Total antioxidant activity of cell aggregates in suspension culture [MS medium supplemented with 10 ${\mu}M$ NAA in the light] was up to 80% of ascorbic acid. By means of HPLC analysis, quantification of the quercetin dehydrate and keamperol profiles from suspension callus was compared. Contents of quercetin dehydrate and keamperol from leaf extracts were 0.07 and 2.24 ${\mu}g/20{\mu}l$, and those from callus 0.56 and 0.18 ${\mu}g/20{\mu}l$, respectively.

• Korean Journal of Acupuncture
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• v.27 no.2
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• pp.95-105
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• 2010

Objectives : The purpose of this study is to investigate the effect of Bacillus-fermented Scutellariae Radix acupuncture solution (SB) on interleukin(IL) production in mouse macrophage stimulatedby lipopolysaccaride(LPS). Methods : Productions of interleukins were measured y High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$(multi-analyte profiling beads) technology. To begin with, cell culture supernatant was obtained after treatment with LPS(1 ${\mu}g/mL$) and SB for 24 hour. Then, it was incubated with the antibody-conj${\mu}g$ated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. After incubating for 30 minutes, Strepavidin-conjugated Phycoerythrin(SAPE) was then added. Incubating for another 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Results : The results of the experiment are as follows. SB significantly inhibited the LPS-induced production of IL-3($9.15{\pm}0.35$ pg/mL) by $6.92{\pm}0.05,\;7.21{\pm}0.11,\;6.96{\pm}0.33,\;and\;7.45{\pm}0.74$ pg/mL at the concentration of 25, 50, 100, and 200 ${\mu}g/mL$ in mouse macrophage RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced production of IL-5($7.30{\pm}0.48$ pg/mL) by $6.50{\pm}0.29,\;6.30{\pm}0.25,\;6.30{\pm}0.25,\;and\;5.80{\pm}0.25$ pg/mL at the concentration of 25, 50 100, and 200 ${\mg}g/mL$ in RAW 264.7 cells (p<0.05). SB significantly inhibited the LPS-induced productiion of IL-9($17.26{\pm}0.19$ pg/mL) by $15.01{\pm}0.43$ pg/mL at the concentration of 25 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced productioh of IL-13($187.80{\pm}2.90$ pg/mL) by $152.80{\pm}4.25,\;172.80{\pm}3.97,\;162.10{\pm}6.67,\;and\;165.30{\pm}11.80$ pg/mL at the concentration fo 25, 50, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-17($18.30{\pm}0.95$ pg/mL) by $13.30{\pm}1.25,\;13.80{\pm}1.11,\;13.30{\pm}0.75,\;and\;14.00{\pm}1.08$ pg/mL at the concentration of 25, 50 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). SB significantly inhibited the LPS-induced production of IL-23($43.90{\pm}0.83$ pg/mL by $39.50{\pm}1.26,\;38.00{\pm}1.78,\;and\;39.60{\pm}2.49$ pg/mL at the concentration of 25, 100, and 200 ${\mu}g/mL$ in RAW 264.7 cells(p<0.05). Conclusions : These results suggest that SB has anti-inflammatory activity related with its inhibition of IL-3, IL-5, IL-13, IL-17, and IL-23 production in macrophages.

• Journal of Life Science
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• v.27 no.12
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• pp.1415-1420
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• 2017

The metallothionein (MT) gene (IbMT3) was selected from an EST library of suspension-cultured sweet potato cells. The MT gene, which is one of abundant ESTs in the library, is involved in stress regulation of cells and tissues. A full-length IbMT3 cDNA was obtained and analysis of its nucleotide sequence revealed that IbMT3 encoded a type 3 MT protein, based on its structural characteristics. The function of type 3 MT in plants is not yet known. Northern blot analysis showed stronger expression of IbMT3 in suspension-cultured cells than in sweet potato plant leaves. Since cell culture is known to impose a state of oxidative stress on cells, sweet potato plants were subjected to oxidative stress to investigate the transcriptional regulation of IbMT3. When the herbicide methyl viologen (MV) was administered for 6, 12, and 24 hr, IbMT3 transcription rapidly increased at 6 hr and then decreased. A cold treatment at $15^{\circ}C$ for 24 and 48 hr resulted in a gradual increase in IbMT3 expression. These findings indicate that IbMT3 expression is regulated in response to environmental and oxidative stress. IbMT3 isoform is expected to have antioxidant effects in sweet potato plants and may play an important role in cellular adaptation to oxidative stress.

• Korean Journal of Plant Tissue Culture
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• v.27 no.3
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• pp.203-211
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• 2000

In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.277-282
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• 1998

Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.