• Title/Summary/Keyword: MTT Assay

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Immuno-Enhancing Effect of Enzymatic Extract of Sargassum coreanum Using Crude Enzyme from Shewanella oneidensis PKA 1008 (Shewanella oneidensis PKA 1008 유래 조효소 처리에 의한 큰잎모자반(Sargassum coreanum) 추출 분해물의 면역증진 효과)

  • Park, Sun-Hee;Kim, Min-Ji;Kim, Go-Eun;Park, So-Yeong;Kim, Koth-Bong-Woo-Ri;Kim, Yeon-Ji;Cho, Young-Je;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.8
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    • pp.919-928
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    • 2017
  • The immuno-enhancing effects of alginate oligosaccharides from Sargassum coreanum were investigated. The alginate oligosaccharides were produced by an alginate-degrading enzyme from S. oneidensis PKA 1008. The degraded alginate oligosaccharides were visualized by thin-layer chromatography developed using a solvent system of 1-butanol/methanol/water, 4:1:2 (v/v/v). Alginate was degraded into dimmers at 60 h. As a result, the levels of Th1 cytokine [interferon $(IFN)-{\gamma}$ and interleukin (IL)-2] and Th2 cytokine (IL-6 and IL-10) increased with increasing incubation time compared to the control in vitro. Enzymatic extract treatment promoted proliferation of splenocytes at concentrations of 100 and 200 mg/kg at 24 h in vivo. Secretion of $IFN-{\gamma}$ and IL-2 significantly increased in a dose-dependent manner at 24 h as well as induced higher production of IgG2a in serum. Natural killer cell activity was measured and tended to increase. In addition, complete blood cell counts increased in a dose-dependent manner. These results indicate that alginate oligosaccharides produced by crude enzyme from S. oneidensis PKA 1008 may have significant immune activities.

Antioxidative and Anti-inflammatory Activity of Extract from Milling By-products of Sorghum Cultivar, 'Hwanggeumchal' (황금찰수수 도정부산물의 항산화 및 항염증 활성)

  • Ra, Ji-Eun;Park, Ji-Young;Seo, Woo Duck;Sim, Eun-Yeong;Ko, Jee Yeon;Nam, Min-Hee;Chung, Ill-Min;Han, Sang-Ik
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.4
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    • pp.463-469
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    • 2014
  • Sorghum has been consumed as one of the important staple food in the semiarid tropics of Africa and Asia. Sorghum is rich in starch, protein, essential vitamins and minerals and grows relatively well in dry climate regions when it compared with other staple food crops. Sorghum has taken an increased interest due to several studies that report about the beneficial effects of sorghum on human health. In the present study, we investigated the antioxidative and activity of extract of milling by-products (hull and bran) of Korean sorghum cultivar, 'Hwanggeaumchal' as well as its grain. Hull extract showed the highest total polyphenol contents ($29.7{\pm}0.2mg\;GAE/100g$) and major four pigments content ($322.6{\pm}14.5mg/100g$). From results of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate (ABTS) radical scavenging activity, hull extract ($IC_{50}$, $6.3{\pm}0.1{\mu}g/mL$) was also showed the strongest antioxidative effects. Bran and grain showed similar polyphenol, pigments contents and antioxidative effects. We determined cell viability by MTT assay and evaluated the anti-inflammatory activity by measuring nitric oxide (NO) of hull, bran and grain methanol extract (0.5% HCl v/v) on RAW 264.7 cells. Hull extract treatment was significantly decreased NO production with dose-dependant manner. Apigeninidin as one of the major pigment of hull was showed inhibitory activity against NO production without cytotoxicitiy. Therefore, sorghum milling by-products can be used as a good source of antioxidative and anti-inflammatory agents.

Anticarcinogenic Effects of Sargassum fulvellum Fractions on Several Human Cancer Cell Lines in vitro (모자반 분획물의 in vitro에서의 항발암효과)

  • 배송자
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.3
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    • pp.480-486
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    • 2004
  • Despite many therapeutic advances in the understanding of the processes in carcinogenesis, overall mortality statistics are unlikely to change until there is reorientation of the concepts for the use of natural products as new anticarcinogenic agents. In this study, we investigated the anticarcinogenic activity, antioxidant and DPPH scavenging activity of Sargassum fulvellum (SF). SF was extracted with methanol, which was further fractionated into five different types: hexane (SFMH), ethylether (SFMEE), ethyl acetate (SFMEA), butanol (SFMB) and aqueous (SFMA) partition layers. We determined the cytotoxic effect of these layers on human cancer cells by MTT assay. Among various partition layers of SF, at starting concentration of 100 $\mu\textrm{g}$/mL, SFMEE showed very high cytotoxicity which were 92, 90 and 84% and kept high throughout 5 concentration levels sparsed by 100 $\mu\textrm{g}$/mL against all three human cancer cell lines: HepG2, HT-29 and HeLa. SFMEA showed a low cytotoxicity at the beginning concentration level, but as the concentration became denser, growth inhibition effect of cancer cell lines started to increase and at 500 $\mu\textrm{g}$/mL, it hit the highest, which were 91, 96 and 98% against the same three cell lines as above. We observed QR induced effect in all fraction layers of SF. SFMEE showed similar tendensy of QR induced effect as did against cytotoxicity. The QR induced effect of SFMEE on HepG2 cells at 25 $\mu\textrm{g}$/mL concentration indicated 3 times higher than the control value of 1.0 and SFMH tended to be concentration-dependent on HepG2 cells. At 100 $\mu\textrm{g}$/mL, the QR induced effects resulted a ratio, which was 2.5 times higher than the control value. In search for antioxidation effects of SF extract and partition layer, the reducing activity on the 1, 1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging potential was sequentially screened. The SFM has similar antioxidant activity as to BHT and vitamin C groups.

Effects of Rhizoma Coptidis on Cellular Activity and IL-6 Production of LPS-treated Periodontal Ligament Cells (황련이 Lipopolysaccharide를 처리한 치주인대세포의 세포활성 및 IL-6 생산에 미치는 영향)

  • Song, Ki-Bum;Kong, Young-Hwan;You, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.26 no.3
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    • pp.641-654
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    • 1996
  • In infectious disease, invasion of host tissue by bacteria or their products frequently induces a wide variety of inflammatory and immunopathologic reaction. Evidence indicates that cytokines are involved in the initiation and progression of chronic inflammatory diseases, such as periodontitis. Interleukin-6, which is a multifunctional cytokine, has important roles in acute and chronic inflammation and may also be implicated in bone resorption. Periodontal diseases are characterized by chronic inflammation of the periodontium with alveolar bone resoption. A principal driving force behind this response appears to lie in the immune system's response to bacteria. Many of the cell components which have been shown to function as virulence factors in gram-negative bacteria are associated with the bacterial surface. Of these, lipopolysaccharide has been characterized as one that mediates a number of biological activities which can lead to the destruction of host tissue. Non-steroidal antiinflammatory drug is used for reduce inflammation, and most of NSAIDs inhibit prostaglandine $E_2$ production, but it is shown that $PGE_2$ production is stimulated by IL-1 in recent study. So, the influence of other cytokines except $PGE_2$ on periodontium can not be avoided. Therefore, new antiinflammatory drug is needed. Rhizoma coptidis is used in oriental medicine for anti-inflammation and antiseptics. In this present study, we examined the IL-6 release in periodontal ligament cells treated with the lipopolysaccharide, and also the effect of rhizoma coptidis on cellular activity and IL-6 production of periodontal ligament cells. To evaluate the effect of rhizoma coptidis on cellular activity, the cells were seeded at a cell density of $1{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 1-6, 10-9 and 10-12 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay were carried out. To evaluate the effect of rhizoma coptidis on IL-6 production, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 10-9 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 3, 6, 12 and 24 hours. Then, amounts of IL-6 production is measured by IL-6 ELISA kit used. The results were as follows : 1. Rhizoma coptidisrbelow to ($10^{-6}g/ml$) significantly increaed cellular activity of periodontal ligament cells than control. 2. Rhizoma coptidist ($10^{-9}g/ml$) significantly increased cellular activity of LPS($5{\mu}g/ml$)-treated periodontal ligament cells than control. 3. LPS(5 and $10{\mu}g/ml$) significantly increased IL-6 production of periodontal ligament cells than control. 4. Rhizoma coptidis($10^{-9}g/ml$) decreased IL-6 production of LPS ($5{\mu}g/ml$)-treated periodontal.ligarnent cells than LPS only tested group. These findings suggest that stimulation of the IL-6 release of periodontal ligament cells by LPS may have a role in the progression of inflammation and alveolar bone resoption in periodontal disease, and that inhibition of the IL-6 release of cells and stimulation of cellular activity by rhizoma coptidis may help the periodontal regeneration.

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The effect of admixture of vitamin $D_3$ and dexamethasone on the activity of osteoblastic cells (Vitamin $D_3$와 Dexamethasone의 복합 투여가 골모세포에 미치는 영향에 관한 연구)

  • Lim, Na-Won;Park, Young Joo;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.29 no.3 s.74
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    • pp.383-397
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    • 1999
  • Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.

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Antioxidaitve and Differentiation Effects of Artemisia capillaris T. Extract on Hydrogen Peroxide-induced Oxidative Damage of MC3T3-E1 Osteoblast Cells (사철쑥(Artemisia capillaris T.) 추출물의 항산화 활성 및 H2O2로 산화적 스트레스를 유도한 조골세포의 활성과 분화에 미치는 영향)

  • Seo, Jee-Eun;Hwang, Eun-Sun;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1532-1536
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    • 2011
  • In this study, the antioxidative activity of Artemisia capillaris T. extract on the proliferation and differentiation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress was investigated in order to determine its protective effect against oxidative stress as well as its availability as an antioxidant material related to treatment of bone diseases. As a result, the total polyphenol content of A. capillaris extract was 90.10 mg/g, whereas the flavonoid content was 4.45 mg/g. A. capillaris extract increased proliferation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress, and also increased the proliferation of differentiated osteoblast cells under oxidative stress. In addition, two differentiation markers, alkaline phosphatase activity and mineralization level, in A. capillaris extract tended to increase. These results indicate that A. capillaris extract suppresses the damage to osteoblasts caused by oxidative stress, which demonstrates its availability as an antioxidant material for preventing bone diseases.

Probiotic Effects of Lactobacillus plantarum Strains Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum 균주들의 프로바이오틱 효과)

  • Lee, Xue-Mei;Lee, Hyun Ah;Kweon, Meera;Park, Eui-Seong;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.12
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    • pp.1717-1724
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    • 2016
  • Probiotic effects of Lactobacillus plantarum pF1 NITE-P1462 (Lp-pF1), L. plantarum KCCM 11352P (Lp-PNU), L. plantarum CBT LP3 KCTC 10782BP (Lp-CB), and L. plantarum KCTC 3099 (Lp-3099) isolated from kimchi and Lactococcus lactis KFCC 11510P (L-lactis) isolated from Doenjang were studied. Resistance to gastric and bile acid, adhesion to intestines in colon cells, thermal stability, and antioxidative and in vitro anticancer effects in HT-29 cancer cells were evaluated. L. plantarum strains showed improved tolerance of gastric and bile acids than L-lactis. Lp-pF1 had better adhesion ability in the intestine than Lp-PNU, Lp-3099, and L-lactis. Lp-pF1 also showed better heat resistance at $50^{\circ}C$, $70^{\circ}C$, and $80^{\circ}C$ than Lp-CB, Lp-3099, and L-lactis. In addition, Lp-pF1 exhibited greater antioxidant activity by scavenging DPPH radicals or hydroxyl radicals and anticancer effects in MTT assay than others. Taken together, these results suggest that L. plantarum isolated from kimchi showed higher probiotic activities with antioxidant and anticancer properties than Lac. lactis isolated from Doenjang. Lp-pF1 revealed the best probiotic activities among L. plantarum and could be used as a promising potential probiotics.

Effects of Schizandra chinensis Extract on the Growth of Intestinal Bacteria Related with Obesity (오미자 추출물이 비만과 관련된 장내 세균의 생육에 미치는 영향)

  • Jeong, Eun-Ji;Lee, Woon-Jong;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.673-680
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    • 2009
  • This study was conducted to screen for plant food materials that improve human intestinal microflora, especially microorganisms associated with obesity. Among 30 tested plant food materials, the extract of Schizandra chinensis inactivated Eubacterium limosum, Bacteroides fragilis and Clostridium spp. Additionally, S. chinensis extract was also found to have a growth-promoting effect on Bifidobacterium spp.. The antimicrobial activity and antioxidant activity of the water extract did not decrease in respond to heating. Additionally, the water extract of S. chinensis did not show a toxic effect on the growth of Caco-2 cells. In vivo feeding tests were performed to investigate the influence of extract on the intestinal microflora in rats. Although the extract did not reduce obesity induced by a high fat diet, it led to significant increase in the population of Bifidobacterium spp. and a decrease in the population of Clostridium spp. in rats. Taken together, these results indicate that S. chinensis could be useful as a functional food component to control intestinal microbial flora.

Gene Expression Profiling of SH-SY5Y Human Neuroblastoma Cells Treated with Ginsenoside Rg1 and Rb1 (Ginsenoside Rg1 및 Rb1을 처리한 신경세포주(SH-SY5Y세포)의 유전자 발현양상)

  • Lee, Joon-Noh;Yang, Byung-Hwan;Choi, Seung-Hak;Kim, Seok-Hyun;Chai, Young-Gyu;Jung, Kyoung-Hwa;Lee, Jun-Seok;Choi, Kang-Ju;Kim, Young-Suk
    • Korean Journal of Biological Psychiatry
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    • v.12 no.1
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    • pp.42-61
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    • 2005
  • Objectives:The ginsenoside Rg1 and Rb1, the major components of ginseng saponin, have neurotrophic and neuroprotective effects including promotion of neuronal survival and proliferation, facilitation of learning and memory, and protection from ischemic injury and apoptosis. In this study, to investigate the molecular basis of the effects of ginsenoside on neuron, we analyzed gene expression profiling of SH-SY5Y human neuroblastoma cells treated with ginsenoside Rg1 or Rb1. Methods:SH-SY5Y cells were cultured and treated in triplicate with ginsenoside Rg1 or Rb1($80{\mu}M$, $40{\mu}M$, $20{\mu}M$). The proliferation rates of SH-SY5Y cells were determined by MTT assay and microscopic examination. We used a high density cDNA microarray chip that contained 8K human genes to analyze the gene expression profiles in SH-SY5Y cells. We analyzed using the Significance Analysis of Microarray(SAM) method for identifying genes on a microarray with statistically significant changes in expression. Results:Treatment of SH-SY5Y cells with $80{\mu}M$ ginsenoside Rg1 or Rb1 for 36h showed maximal proliferation compared with other concentrations or control. The results of the microarray experiment yielded 96 genes were upregulated(${\geq}$3 fold) in Rg1 treated cells and 40 genes were up-regulated(${\geq}$2 fold) in Rb1 treated cells. Treatment with ginsenoside Rg1 for 36h induced the expression of some genes associated with protein biosynthesis, regulation of transcription or translation, cell proliferation and growth, neurogenesis and differentiation, regulation of cell cycle, energy transport and others. Genes associated with neurogenesis and neuronal differentiation such as SCG10 and MLP increased in ginsenoside Rg1 treated cells, but such changes did not occur in Rb1-group. Conclusion:Our data provide novel insights into the gene mechanisms involved in possible role for ginsenoside Rg1 or Rb1 in mediating neuronal proliferation or cell viability, which can elicit distinct patterns of gene expression in neuronal cell line. Ginsenoside Rg1 have more broad and strong effects than ginsenoside Rb1 in gene expression and related cellular physiology. In addition, we suggest that SCG10 gene, which is known to be expressed in neuronal differentiation during development and neuronal regeneration during adulthood, may have a role in enhancement of activity dependent synaptic plasticity or cytoskeletal regulation following treatment of ginsenoside Rg1. Further, ginsenoside Rg1 may have a possible role in regeneration of injured neuron, promotion of memory, and prevention from aging or neuronal degeneration.

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Biological Activities of Isolated Icariin from Epimedium koreanum Nakai (삼지구엽초로부터 분리한 Icariin의 생리활성)

  • Kim, Seo-Jin;Park, Myoung-Su;Ding, Tian;Wang, Jun;Oh, Deog-Hwan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.10
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    • pp.1397-1403
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    • 2011
  • Epimedium koreanum Nakai is a wild medicinal plant commonly consumed in South Korea due to its health beneficial effects. In the present study, the antioxidative, antimutagenic and immunological activities of E. koreanum Nakai extracts were investigated for their use in food. The yields of icariin compounds from the ethanol extract as well as the ethyl acetate, butanol, hexane, water, and chloroform fractions of E. koreanum were 27.9, 2.5, 1.7, 1.4, and 1.3 ${\mu}g/g$, respectively. The icariin components (295.5 ${\mu}g/g$) were collected from the ethyl acetate fraction by thin layer chromatography (TLC) and analyzed via high performance liquid chromatography (HPLC). The antioxidant activities of each fraction were as follows: ethyl acetate (49.0 ${\mu}g/mL$), butanol (59.2 ${\mu}g/mL$), hexane (119.8 ${\mu}g/mL$), water (122.0 ${\mu}g/mL$), and chloroform (138.5 ${\mu}g/mL$), based on $RC_{50}$ ${\mu}g/mL$. Icariin, isolated and identified as the main component, showed strong antioxidant activity with a $RC_{50}$ value of 15.3 ${\mu}g/mL$, which was higher than those of ascorbic acid (19.5 ${\mu}g/mL$) and ${\alpha}$-tocopherol (18.2 ${\mu}g/mL$). In an Ames test, none of the fractions produced mutagenic effects on Salmonella Typhimurium TA98 and TA100. In an immunomodulating activity test, the effects of E. koreanum Nakai on B cells (Rhamos) and T cells (Jurkat) were investigated. These results show that the growth and viability of B and T cells were increased by isolated icariin components for 1.27 and 1.28 fold, respectively. These results also provide preliminary data for the development of E. koreanum Nakai as an edible food material.