Anti-diabetic effects of extracts and fractions of Sasa borealis (SB), white lotus roots (LR) and leaves (LL), and their mixture were determined in 3T3-L1 adipocytes and Min6 cells by investigating insulin-sensitizing activity and glucose-stimulated insulin secretion, respectively. SB, LR, LL, and mixture of SB, LR, and LL (3 : 2 : 3) were extracted using 70% ethanol, and m mixture extract was fractionated by XAD-4 column chromatography with serial mixture solvents of methanol and water. Fractional extractions were utilized for anti-diabetic effect assay. SB and LR extracts increased insulin-stimulated glucose uptake, but not as much as mixture of SB, LR, and LL. Significant insulin-sensitizing activities of 20 and 80% methanol fractions of SB, LR, and LL mixture extract were observed in 3T3-L1 adipocytes, giving 0.5 or $5\;{\mu}g/mL$ each fraction with 0.2 nM insulin to attain glucose uptake level similar to that attained by 10 nM insulin alone. Similar to pioglitazone, peroxisome proliferators-activated $receptor-{\gamma}\;(PPAR-{\gamma})$ agonist, 20 and 80% methanol fractions increased adipocytes by stimulating differentiation from fibroblasts and triglyceride synthesis. LL extract and 20, 60, and 80% methanol fractions of the mixture suppressed ${\alpha}-amylase$ activity, but did not modulate insulin secretion capacity of Min6 cells in both low and high glucose media. These data suggest 20 and 80% methanol tractions contain potential insulin sensitizers with functions similar to that of $PPAR-{\gamma}$ agonist. Crude extract of SB, LR, and LL mixture possibly improves glucose utilization by enhancing insulin-stimulated glucose uptake and inhibiting carbohydrate digestion without affecting insulin secretion in vivo.
Kim, H.K.;Lim, Jung-Dae;Hyun, Tae-Kyung;Lee, Hyeon-Yong;Lee, Jin-Ha;Yu, Chang-Yeon
Korean Journal of Medicinal Crop Science
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v.9
no.4
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pp.310-317
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2001
The regenerated-bulblets placed in liquid free media resulted in good formation of roots and bulblets. On 1/4 MS free medium, roots and bulblets were predominantly induced. The 1/4 MS liquid medium supplemented with plant growth regulators was the best suitable condition for elongation of leaves and roots. Somatic embryos were frequently developed from embryogenic callus in liquid media with 2,4-D 1mg/ l . On free liquid media, the viability of callus reduced. As the salt strength of MS media reduces, the viability of callus reduced significantly. However, Leaves were induced from several callus clumps. When leaves, roots and bulb-scale segments were placed on MS media containing NAA 1mg/ l or 2,4-D 1mg/ l and various sucrose concentration, the best result about the differentiation, growth of leaf and the differentiation of leaf was obtained on MS media added 1.5% sucrose and 2,4-D 1mg/ l, 3% sucrose and NAA 1mg/ l, and 1.5% sucrose and NAA 1mg/ l, respectively. Also the better result differentiation, growth of root and differentiation of bulb was obtained on MS media with 6% sucrose and NAA 1mg/ l. Spermidine promoted the growth of leaf and the differentiation of bulb. However, spermine promoted the differentiation of leaf, the differentiation and the growth of root in MS solid media. On the MS liquid media, both spermine and spermidine stimulated organogenesis from bulb-scale segments. Regenerated plantlets were acclimatizated and grown in greenhouse in vermiculite + perlite (1 : 1 by volume) well. The optimal soil condition of rooting for plantlets regenerated was in peat moss.
Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.Multiple shoots obtained in MS medium suppler with 5.0 mg/L BA though shoot-tip culture. The frequency of vitrified shoot was lower on Bacto-agar medium than on Gelrite as gelling agent. Addition of activated charcoal at concentrations of 0.1~0.3% reduced vitrification and markedly increased shoot growth, and formation and growth of roots, but significantly reduced the number of shoots formed. The ratio of fresh weight to dry weight was decreased by increasing light intensity and agar concentration. Eight-tenths times of macroelement of MS medium was observed to be effective for shoot formation. Addition of IAA effectively promoted shoot formation in both shoot tip and node-bud explants. Supplement of 5.0 mg/L BA, 0.3 mg/L IAA to MS medium was most effective in shoot proliferation on shoot tip and node-bud explants.
Whole plants were regenerated from callus induced from leaf explants in hybrid kiwi (Actinidia deliciosa${\times}$A. arguta). Callus was induced from leaf explants which cultured on MS solid medium supplemented with combination of auxin (2,4-D, NAA: 0.1~0.5 mg/l) and cytokinin (BA: 0.1~0.2 mg/l). them, the highest callus formation (96.2%) was obtained from the treatment of 0.5 mg/1 2,4-D+0.1 mg/l NAA+0.05 mg/l BA. In the experiment of adventitious shoots induction from primary shoots, only a few shoots were produced in the treatment of 1.0 mg/l BA+0.05 mg/l IBA or 2.0 mg/l BA+0.05 mg/l lBA. As the callus were transferred to the secondary shoot-inducing medium, multiple shoots were obtained from the medium supplemented with 1.0, 2.0 or 5.0 mg/l zeatin in addition to the mixed treatments of BA, thidiazuron (TDZ) or zeatin. However, no multiple shoots were induced on the BA-contained medium of concentrations. Therefore it turned out that addition of BA to medium was less effective for induction of multiple shoots from callus in Actinidia deliciosa${\times}$A. arguta. For producing adventitious roots from shoots, the best frequency of rooting (83.3%) were recorded on the treatment of in vitro rooting (Standardi (St)+1.0 mg/l IBA). On the other side, the lowest result (40.0%) were shown in the treatment of 500 mg/l IBA, 1 hr. Whole plants with shoots and roots were recovered and acclimatized successfully.
Lee, Joung Kwan;Kwon, Young Hee;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Jeong, Mi Jin;Son, Sung Won;Suh, Gang Uk
Proceedings of the Plant Resources Society of Korea Conference
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2019.04a
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pp.53-53
/
2019
Cypripediums, popularly called lady's slippers or moccasin flowers, are the showiest and most sought after hardy terrestrial orchids, collected and grown by orchid and alpine plant enthusiasts alike. In Korea, 4 species of cypripedium are reported as Cypripedium japonicum, C. macranthos, C. guttatum, and C. calceolus. We had already reported the feasibilities of C. macranthos and C. guttatum with in vitro germination methods from immature seeds. The seeds of white lady's slipper orchid (Cypripedium macranthos Sw. alba) were collected 65 days after pollination in 2018. The green pods were sterilized with flame and sowed immediately on the POM(Phytomax orchid maintenance media(R), Sigma) supplemented with BAP 0, 0.5, 1.0 mg/L and NAA 0, 1, 2mg/L. The germination of seed was observed 90 days after sowing, and the plantlets were subcultured to the same media according to the size of the protocorm with 1~2, 2~3, 3~4, 5~6, 7~8mm. The time of the subculture to the new media seems to be critical factors of forming rhizoids which is the hairy root of the cypripediums. As a results, the protorms of the white lady's slipper orchid was successfully germinated in the POM media supplemented BAP 0.5 and NAA 1.0 mg/L. The roots and rhizoids were formed in 5~6mm protocorms subculture over 95% survival ratio. We also tried to subculture to liquid medium without activated charcoal, however the browning or malformation of the roots was observed in the root. The formation of shoots from the protocorm was effectively enhanced in the POM media with non-additives of plant growth regulators. These results indicate the possibility of high and stable production and practical industrialization of endangered white lady's slipper orchids.
Chung, Eu Jin;Hossain, Mohammad Tofajjal;Khan, Ajmal;Kim, Kyung Hyun;Jeon, Che Ok;Chung, Young Ryun
The Plant Pathology Journal
/
v.31
no.2
/
pp.152-164
/
2015
Biological control of major rice diseases has been attempted in several rice-growing countries in Asia during the last few decades and its application using antagonistic bacteria has proved to be somewhat successful for controlling various fungal diseases in field trials. Two novel endophytic Bacillus species, designated strains YC7007 and $YC7010^T$, with antimicrobial, plant growth-promoting, and systemic resistance-inducing activities were isolated from the roots of rice in paddy fields at Jinju, Korea, and their multifunctional activities were analyzed. Strain YC7007 inhibited mycelial growth of major rice fungal pathogens strongly in vitro. Bacterial blight and panicle blight caused by Xanthomonas oryzae pv. oryzae (KACC 10208) and Burkholderia glumae (KACC 44022), respectively, were also suppressed effectively by drenching a bacterial suspension ($10^7cfu/ml$) of strain YC7007 on the rhizosphere of rice. Additionally, strain YC7007 promoted the growth of rice seedlings with higher germination rates and more tillers than the untreated control. The taxonomic position of the strains was also investigated. Phylogenetic analyses based on 16S rRNA gene sequences indicated that both strains belong to the genus Bacillus, with high similarity to the closely related strains, Bacillus siamensis KACC $15859^T$ (99.67%), Bacillus methylotrophicus KACC $13105^T$ (99.65%), Bacillus amyloliquefaciens subsp. plantarum KACC $17177^T$ (99.60%), and Bacillus tequilensis KACC $15944^T$ (99.45%). The DNA-DNA relatedness value between strain $YC7010^T$ and the most closely related strain, B. siamensis KACC $15859^T$ was $50.4{\pm}3.5%$, but it was $91.5{\pm}11.0%$ between two strains YC7007 and $YC7010^T$, indicating the same species. The major fatty acids of two strains were anteiso-$C_{15:0}$ and iso $C_{15:0}$. Both strains contained MK-7 as a major respiratory quinone system. The G+C contents of the genomic DNA of two strains were 50.5 mol% and 51.2 mol%, respectively. Based on these polyphasic studies, the two strains YC7007 and $YC7010^T$ represent novel species of the genus Bacillus, for which the name Bacillus oryzicola sp. nov. is proposed. The type strain is $YC7010^T$ (= KACC $18228^T$). Taken together, our findings suggest that novel endophytic Bacillus strains can be used for the biological control of rice diseases.
Kim, Jin-Cheol;Kim, Mi-Ri;Ko, Hyun-Jung;Yang, Won-Kyung
Restorative Dentistry and Endodontics
/
v.34
no.4
/
pp.371-376
/
2009
We evaluated in vitro microleakage of Mineral Trioxide Aggregate (MTA) powder with 4-methacryloxyethyl trimellitate anhydride (4-META) / methyl methacrylate (MMA) & tri-n-butylborane (TBB) resin as a retrograde filling material by using methylene blue dye method. Fifty-two single rooted, extracted teeth were instrumented and obturated with gutta percha and AH plus sealer. The apical 3mm of each root was resected and 3mm deep ultrasonic root end preparation was done. External surface of roots was coated with nail varnish. Prepared teeth were randomly divided into five groups; Negative control: completely covered with nail varnish; Positive control: coated with nail varnish except for apical foramen; Group 1 (retrofilled with Portland cement); Group 2 (retrofilled with MTA); Group 3 (retrofilled with MTA powder mixed with 4-META/MMA & TBB resin). Immediately after completion of root-end filling, all specimens were submerged in methylene blue dye for 72 hours in $37^{\circ}C$incubator. The roots were longitudinally sectioned and measured for extent of dye penetration by three different examiners under microscope (${\times}$10). The results were statistically analyzed using one way ANOVA and Turkey's HSD test. No leakage was evident in negative control and complete leakage in positive control group. Group 3 showed significantly less leakage than group 1 and 2 (p < 0.01). There was no significant difference between group 1 and 2 (p > 0.01). It was concluded that MTA powder with 4-META/MMA & TBB resin was excellent in reducing initial apical microleakage.
Inhibitory effects of Maclura amboinenesis Bl, one plant used traditionally for the treatment of cancers, on metastatic potential of highly metastatic B16F10 melanoma cells were investigated in vitro. Cell proliferation was assessed using the MTT colorimetric assay. Details of metastatic capabilities including invasion, migration and adhesion of B16F10 melanoma cells were examined by Boyden Chamber invasion and migration, scratch motility and cell attachment assays, respectively. The results demonstrated that n-hexane and chloroform extracts exhibited potent anti-proliferative effects (p<0.01), whereas the methanol and aqueous extracts had less pronounced effects after 24 h exposure. Bioactivity-guided chromatographic fractionation of both active n-hexane and chloroform extracts led to the isolation of two main prenylated xanthones and characterization as macluraxanthone and gerontoxanthone-I, respectively, their structures being identified by comparison with the spectral data. Interestingly, both exhibited potent effective effects. At non-toxic effective doses, n-hexane and chloroform extracts (10 and $30{\mu}g/ml$) as well as macluraxanthone and gerontoxanthone-I (3 and $10{\mu}M$) significantly inhibited B16F10 cell invasion, to a greater extent than $10{\mu}m$ doxorubicin, while reducing migration of cancer cells without cellular cytotoxicity. Moreover, exposure of B16F10 melanoma cells to high concentrations of chloroform ($30{\mu}g/ml$) and geratoxanthone-I ($20{\mu}M$) for 24 h resulted in delayed adhesion and retarded colonization. As insights into mechanisms of action, typical morphological changes of apoptotic cells e.g. membrane blebbing, chromatin condensation, nuclear fragmentation, apoptotic bodies and loss of adhesion as well as cell cycle arrest in the G1 phase with increase of sub-G1 cell proportions, detected by Hoechst 33342 staining and flow cytometry were observed, suggesting DNA damage and subsequent apoptotic cell death. Taken together, our findings indicate for the first time that active n-hexane and chloroform extracts as well as macluraxanthone and gerontoxanthone-I isolated from Maclura amboinensis Bl. roots affect multistep of cancer metastasis processes including proliferation, adhesion, invasion and migration, possibly through induction of apoptosis of highly metastatic B16F10 melanoma cells. Based on these data, M. amboinensis Bl. represents a potential candidate novel chemopreventive and/or chemotherapeutic agent. Additionally, they also support its ethno-medicinal usage for cancer prevention and/or chemotherapy.
The antioxidant and photoprotective effects of various extracts from the roots of Rumex crispus L. were evaluated. The concentrations ($IC_{50}$) of various extracts required to exert a 50% reducing effect on a DPPH radical were found to be 0.005~0.093 mg/mL. The ethyl acetate extract showed a more remarkable effect than the positive control ascorbic acid. The concentrations ($QC_{50}$) of the butanol and ethyl acetate extracts required to exert a 50% reducing effect on the singlet oxygen $^1O_2$ were found to be 0.464 and 0.365 mg/mL, respectively. Both extracts were also found to protect the in vitro biological system from the detrimental effect of a singlet oxygen $^1O_2$ on type II photosensitization in E. coli and genomic DNA. Among all the tested extracts, the ethyl acetate and butanol extracts contained higher amounts of total phenolic contents. The results suggest that our study may contribute to the development of new bioactive products with potential applications to the reduction of photo-produced oxidative stress involving reactive oxygen species in living organisms.
Kim, Kyung-Min;Park, Min-Hee;Kim, Kyung-Hee;Im, Sang-Hyun;Park, Yoo-Hwa;Kim, Young-Nam
Journal of Applied Biological Chemistry
/
v.52
no.2
/
pp.58-64
/
2009
Total polyphenol contents and antioxidative activity of water and ethanol extracts from Sea Buckthorn (Hippophae rhamnoides) leaves, branches (bough and twig) and roots were analyzed. The level of crude ash and crude protein in water extract of leaves, branches (bough and twig), and roots were shown to be a bit higher than ethanol extracts. Especially crude protein contents from water extract of leaves, bough, twig, and roots were 14.90, 18.60, 18.03, and 16.61% respectively. Total polyphenol content of ethanol extracts of all parts of Sea buckthorn was ranged from 106.33${\pm}$2.32 ${\mu}g/g$ to 147.78${\pm}$3.06 ${\mu}g/g$ showing higher amount than water extracts. To investigate antioxidative activity of Sea buckthorn, DPPH free radical scavenging activity, hydroxy radical scavenging activity, and SOD-like activity were analyzed. The results showed that the antioxidative activity of ethanol extracts was relatively higher than water extracts. The $IC_{50}$ values for DPPH free radical scavenging activity was ranged from 23.58${\pm}$0.84 ${\mu}g/mL$ to 59.35${\pm}$1.69 ${\mu}g/mL$. Compared to 68.85${\pm}$1.44% of SOD-like activity from L-ascorbic acid used as a control, the ethanol extract of Sea buckthorn branches showed relatively strong activity of 35.03${\pm}$2.33%. The highest hydroxy radical scavenging activity was shown as 66.12${\pm}$8.73% from ethanol extract of Sea buckthorn roots which was similar value to 72.47${\pm}$2.83% of L-ascorbic acid.
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