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Antioxidative, Antimicrobial and Cytotoxic activities of Fagopyrum esculentum $M{\ddot{o}}ench$ Extract in Germinated Seeds (발아 메밀 추출물의 항산화.항균활성 및 세포독성)

  • Hwang, Eun-Ju;Lee, Sook-Young;Kwon, Su-Jung;Park, Min-Hee;Boo, Hee-Ock
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.1
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    • pp.1-7
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    • 2006
  • This research was conducted to investigate the possibilities of usage of germinated-buckwheat (Fagopyrum esculentum $M{\ddot{o}}ench$) by examining antioxidative, antimicrobial and cytotoxic effects of extracts from different germinated root length of buckwheat. Antioxidant activity $(RC_{50})$ was shown higher in extracts of non-germinated seed $(50.41\;{\mu}g/mL)$ and root length 10 mm $(80.57\;{\mu}g/mL)$, 2 mm $(93.77\;{\mu}g/mL)$, 5 mm $(107.09\;{\mu}g/mL)$ than BHT $(163.96\;{\mu}g/mL)$ as a synthetic antioxidant. In antimicrobial activity, non-germinated and germinated seeds were formed inhibitory zone against S. aureus $(4{\sim}10\;mm)$, P. aeruginosa $(2{\sim}9\;mm)$ at the concentrations of $10{\sim}40\;mg/mL$ but B. subtilis, E. coli and S. typhimurium were not apparent antimicrobial activity. Extracts of germinated seed also decreased their antimicrobial activity compared to non-germinated seed extract. In addition, the growth of Calu-6 was inhibited of both 5 mm root length germinated and non-germinated seeds $(800\;{\mu}g/mL)$ as 95.12% and 87.15%, respectively, but these did not show any influence on cytotoxic effect against MCF-7 and Caco-2 cell lines. Extracts of 2 mm and 5 mm germinated seeds were also inhibited against Calu-6 and SNU-601 cell lines.

Development of Natural Antioxidants and Whitening Agents for Cosmeceuticals

  • Kim, Jong-Pyung
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2007.11a
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    • pp.79-92
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    • 2007
  • Oxidative stress have known to be a risk factor for the degenerative processes and closely related to a lot of diseases. It is well established that antioxidants are good in protection and therapeutic means against oxidative damage. There is increasing interest in natural antioxidants and many natural antioxidants have been found and utilized as the possible protection for various diseases and skin aging. We have screened natural antioxidant agents for cosmeceuticals, nutraceuticals, and drugs as therapeutic and preventive means against oxidative stress, and have developed a number of novel antioxidants from various natural sources. A novel melanin synthesis inhibitor, Melanocin A, isolated from the metabolite of a fungal strain Eupenicillium shearii F80695 inhibited mushroom tyrosinase and melanin biosynthesis of B16 melanoma cells with $IC_{50}$ value of 9.0 nM and MIC value of $0.9\;{\mu}M$, respectively. Melanocin A also exhibited potent antioxidant activity by scavenging of DPPH and superoxide anion radicals. UV was found to increase the level of hydrogen peroxides and other reactive oxygen species (ROS) in skin tissues. This increase in ROS may not only alter the structure and function of many genes and proteins directly but may also modulate their expressions through signal transduction pathways and, ultimately, lead to skin damage. We investigated the effect of Melanocin A on UV-induced premature skin aging. Firstly, the effect of Melanocin A on UV-induced matrix metalloproteinase (MMP)-9 expression in an immortalized human keratinocyte cell line, HaCaT in vitro was investigated. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and Melanocin A suppressed this expression in a dose-dependent manner. We then investigated UV-induced skin changes in hairless mice in vivo by Melanocin A. Chronic exposure of hairless mouse dorsal skin to UV increased skin thickness and induced wrinkle formation and the gelatinase activities of MMP-2 and MMP-9. Moreover, Melanocin A significantly suppressed UV-induced morphologic skin changes and MMP-2 and MMP-9 expression. These results show that Melanocin A can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that Melanocin A should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. Terrein is a bioactive fungal metabolite isolated from Penicillium species. Terrein has a relatively simple structure and can be easily synthesized. However, the biologic effects of terrein are comparatively unknown. We found for the first time that terrein potently inhibit melanin production in melanocytes and has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 mM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrain treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrain reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

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Optimization of Large Scale Culture Conditions of Bacillus ehimensis YJ-37 Antagonistic to Vegetables Damping-off Fungi (채소류 모잘록병균에 길항하는 Bacillus ehimensis YJ-37의 대량배양 최적조건)

  • 주길재;김진호
    • Journal of Life Science
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    • v.12 no.3
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    • pp.242-249
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    • 2002
  • The optimal culture conditions in 500$m\ell$ flask suture, 5$\ell$ jar fermenter and 2,000 $\ell$ large stale culture were investigated to maximize the production of antibiotic on Rhizoctonia solani AC4, the causal agent of vegetables damping-off, by the strain Bacillus ehimensis YJ-37. Starch (1.5%) as a carbon source, peptone (0.6%) as a nitrogen source and MgC1$_2$(0.15%) as a metal ion in the medium containing N $a_2$HP $O_4$(0.3%) showed the highest production of the antibiotic(s) in a rotary shake (200 rpm). Optimal initial pH of the culture medium, culture temperature and culture time for the antibiotic(s) production were pH 8.0, 32$^{\circ}C$ and 54hrs, respertively. Under the optimal renditions in flask culture, cell growth and antifungal activity (clear zone size) were 1.42 $\times$ 10$^{8}$ cfu/$m\ell$ (21g-DCW/ $\ell$) and 13.9 mm, respectively. In 5 $\ell$ jar fermenter (medium 3 $\ell$), optimal air flow, agitation speed and culture time for the antibiotic(s) production were 2 vvm, 200 rpm and 48hrs, respectively. Under the optimal conditions in 5 $\ell$ jar fermenter, tell growth and antifungal activity were 2.06 $\times$ 10$^{8}$ cfu/$m\ell$ (30g-DCW/ $\ell$) and 13.4 mm, respectively. Under the culture conditions of air flow (30 vvm) and agitation speed (200 rpm) at 32$^{\circ}C$ for 10 days in 2,000 $\ell$ large scale culture (medium 1,800 $\ell$, pH 8.0), cell growth and antifungal activity were 0.81$\times$10$^{8}$ cfu/$m\ell$ (12g-DCW/ $\ell$) and 8.6 mm, respectively.

Effects of the Glycoprotein Isolated from Pteridium aquilinum on the Immune Function of Mice (고사리 단백다당(Pteridium aquilinum Glycoprotein, PAG)이 마우스 면역활성에 미치는 영향)

  • Park, Hyeon-Ae;Kweon, Mee-Hyang;Han, Hyung-Mee;Sung, Ha-Chin;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.976-982
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    • 1998
  • The effects of the glycoprotein (PAG) isolated from Pteridium aquilinum on the immune function was examined in mice. PAG was intraperitoneally administered into BALB/C mice for 14 days and the antibody forming ability to hen egg lysozyme (HEL) and the blastogenic responses of splenocytes were measured. PAG treatment significantly increased antibody formation to HEL in a dose-dependent manner. Blatogenesis of splenocytes in response to lipopolysaccharide (LPS, B-cell specific mitogen) or phytohemagglutinin (PHA, T-cell specific mitogen) was also increased after treatment with PAG, indicating that the PAG increases both humoral and cellular immunities. To examine whether the immune function of PAG was via a direct effect on the lymphocytes, splenocytes were isolated from BALB/C mice, exposed to various concentrations of PAG in vitro and the blastogenic responses were measured. In vitro exposure to PAG significantly increased blastogenesis of splenocytes to LPS up to $500{\;}{\mu}g/kg$, whereas the blastogenic response to PHA was not altered by PAG treatment. To identify the fraction responsible for the increase in the immune function, the effect of periodate digest, pronase digest or purified polysaccharide on the antibody production to HEL was examined. Crude protein fraction of PAG significantly increased the antibody formation to HEL. On the other hand, both crude and purified polysaccharide fractions did not have any effects on the antibody production ability. These data indicated that 1) PAG increased both humoral and cellular immune functions, 2) the increase in humoral immunity was probably via a direct action of PAG on lymphocytes and 3) the protein portion of PAG was responsible for the increase in humoral immunity.

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Biological Activities and Physiochemical Properties of Gangwon-do Endemic Makjang Products (강원도 시판 막장제품의 이화학적 품질특성 및 생리활성 조사)

  • Kim, Byoung-Mok;Jung, Jee-Hee;Lim, Ji-Hoon;Jung, Min-Jeong;Jeong, Jae-Whung;Choi, Yong-Suck;Sim, Jea-Man;Jeong, In-Hak;Kim, Young-Myoung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.6
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    • pp.862-873
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    • 2015
  • In this study, we investigated the physiochemical properties and biological activities of Gangwon-do endemic Makjang (MJ) products (12 types). The pH levels of all samples were in the range of 4.43 to 5.69, and MJ5 showed the highest pH (5.69). The salinities of all samples ranged from 11.1% to 16.9%. Hunter color values for L (lightness), a (redness), and b (yellowness) ranged from 26.2 to 36.9, 3.9 to 11.5, and 6.5 to 16.6, respectively. The amino nitrogen content of MJ2 was highest, whereas the total content of free amino acids of MJ11 (4,657.7 mg%) was highest. Total fatty acid contents of all samples ranged from 1,598.6 mg% to 2,874.4 mg%, with MJ10 showing the highest fatty acid content. The content of total polyphenolic compounds ranged from 401.48 to $746.67{\mu}g$ tannic acid equivalent/mL, with MJ11 showing the highest content. The 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis(3-ethylbenzothiozoline-6-sulfonic acid) radical scavenging effects of MJ11, MJ8, and MJ4 were 51.30% and 82.5%, 41.29% and 67.0%, and 49.88% and 87.7%, respectively. MJ12 showed the strongest growth inhibitory effect on lung cancer A549 cells, whereas MJ5 showed the strongest growth inhibitory effect on AGS gastric cancer cell and MCF-7 breast cancer cell. MJ7 showed greater lipid accumulation inhibitory activity in HepG2 cells than the others. ACE inhibitory activity of MJ11 was the highest among the samples.

Protective Effect of the Ethyl Acetate-fraction of Methanol Extract of Ophiophogon japonicus on Amyloid beta Peptide-induced Cytotoxicity in PC12 Cells (소엽맥문동-에틸아세테이트 분획물의 아밀로이드 베타단백질-유발 세포독성에 대한 억제 효능)

  • Moon, Ja-Young;Kim, Eun-Sook;Choi, Soo-Jin;Kim, Jin-Ik;Choi, Nack-Shik;Lee, Kyoung;Park, Woo-Jin;Choi, Young-Whan
    • Journal of Life Science
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    • v.29 no.2
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    • pp.173-180
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    • 2019
  • Amyloid ${\beta}$-protein ($A{\beta}$) is the principal component of senile plaques characteristic of Alzheimer's disease (AD) and elicits a toxic effect on neurons in vitro and in vivo. Many environmental factors, including antioxidants and proteoglycans, modify $A{\beta}$ toxicity. It is worthwhile to isolate novel natural compounds that could prove therapeutic for patients with AD without causing detrimental side effects. In this study, we investigated the in vitro neuroprotective effects of the ethyl acetate fraction of methanol extract of Ophiophogon japonicas (OJEA fraction). We used an MTT reduction assay to detect protective effects of the OJEA fraction on $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells. We also used a cell-based ${\beta}$-secretase assay system to investigate the inhibitory effect of the OJEA fraction on ${\beta}$-secretase activity. In addition, we performed an in vitro lipid peroxidation assay to evaluate the protective effect of the OJEA fraction against oxidative stress induced by $A{\beta}_{25-35}$ in PC12 cells. The OJEA fraction had strong protective effects against $A{\beta}_{25-35}$-induced cytotoxicity to PC12 cells and was strongly inhibitory to ${\beta}$-secretase activity, which resulted in the attenuation of $A{\beta}$ generation. In addition, the OJEA fraction significantly decreased malondialdehyde (MDA) content, which is induced by the exposure of PC12 cells to $A{\beta}_{25-35}$. Our results suggested that the OJEA fraction contained active compounds exhibiting a neuroprotective effect on $A{\beta}$ toxicity.

Anti-inflammatory and Cytotoxic Screening Evaluation of Macroalgae Resources (국내 해조류 자원의 항염증 및 세포독성 스크리닝 평가)

  • Kim, C.W.;Chang, K.J.;Kim, Y.B.;Kim, D.H.;Chae, C.J.;Choi, H.G.;Koo, H.J.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.22 no.2
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    • pp.69-79
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    • 2020
  • In this study, the anti-inflammatory and cytotoxic effects of hot-water extracts from 10 kinds of macroalgae in Korea were investigated. It was selected materials in consideration of biological activity and industrial potential as follows: Caulerpa okamurae; Codium fragile; Ulva australis; Ishige foliacea; Saccharina japonica; Sargassum horneri; Undaria pinnatifida; Gloiopeltis tenax; Gracilaria verrucosa; Porphyra tenera. Results showed that S. japonica and G. tenax significantly decreased NO productionn in LPS-stimulated Raw 264.7 cells at concentrations of 100, 1000 ㎍/mL and 1000 ㎍/mL, respectively. However, most of the other macroalgae used in the experiment did not affect NO production. It was observed that all macroalgae extracts except for the highest concentration (1000 ㎍/mL) treatment group of P. tenera did not affect the viability in Raw 264.7 cells. In addition, there was not significant decrease in cell viability by macroalgae extracts treatment in HINAE cells. These results suggest that S. japonica and G. tenax could be used as potential safe natural anti-inflammatory agents for food and feed additives. Also, the results of this study are expected to be used as basic data for the development of functional materials for 10 kinds of macroalgae resources in Korea.

Discovery of UBE2I as a Novel Binding Protein of a Premature Ovarian Failure-Related Protein, FOXL2 (조기 난소 부전증 유발 관련 단백질인 FOXL2의 새로운 결합 단백질 UBE2I의 발견)

  • Park, Mira;Jung, Hyun Sook;Kim, Hyun-Lee;Pisarska, Margareta D.;Ha, Hye-Jeong;Lee, Kangseok;Bae, Jeehyeon;Ko, Jeong-Jae
    • Development and Reproduction
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    • v.12 no.3
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    • pp.289-296
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    • 2008
  • BPES (Blepharophimosis/Ptosis/Epicanthus inversus Syndrome) is an autosomal dominant disorder caused by mutations in FOXL2. Affected individuals have premature ovarian failure (POF) in addition to small palpebral fissures, drooping eyelids, and broad nasal bridge. FOXL2 is a member of the forkhead family transcription factors. In FOXL2-deficient ovaries, granulosa cell differentiation dose not progress, leading to arrest of folliculogenesis and oocytes atresia. Using yeast two-hybrid screening of rat ovarian cDNA library with FOXL2 as bait, we found that small ubiquitin-related modifier (SUMO)-conjugating E2 enzyme UBE2I protein interacted with FOXL2 protein. UBE2I also known as UBC9 is an essential protein for processing SUMO modification. Sumoylation is a form of post-translational modification involved in diverse signaling pathways including the regulation of transcriptional activities of many transcriptional factors. In the present study, we confirmed the protein-protein interaction between FOXL2 and UBE2I in human cells, 293T, by in vivo immunoprecipitation. In addition, we generated truncated FOXL2 mutants and identified the region of FOXL2 required for its association with UBE2I using yeast-two hybrid system. Therefore, the identification of UBE2I as an interacting protein of FOXL2 further suggests a presence of novel regulatory mechanism of FOXL2 by sumoylation.

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Development of heat exchanger for underground water heat. II - Design and manufacture for heat exchanger of underground water - (지하수 이용을 위한 열교환기 개발. II - 지하수이용 냉·난방기 설계제작 -)

  • Lee, W.Y.;Ahn, D.H.;Kim, S.C.;Park, W.P.;Kang, Y.G.;Kim, S.B.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.4 no.1
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    • pp.128-137
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    • 2002
  • This study was conducted to develop the heat exchanger by utilizing the heat energy of underground water(15℃), which might be used for cooling and heating system of the agricultural facilities. We developed the heat exchanger by using the parallel type plat fin tube made of Aluminum(Al 6063), which was named Aloo-Heat(No. 0247164, offered by Korean Intellectual property Office). The trial manufactures were made from Aloo-heat which was 600mm, 700mm length respectively, and It were welded to the end "U" type in order to direct flow of the underground water. The performance test was carried out under the condition of open space and room temperature with the change of flow rate of the underground water and air. The results are as follows. 1. The trial manufactures had convection heat value from 33 to 156 W/m2℃, and It was coincided with design assumption. 2. The amount of energy transfer was increased with the increment of the area of heat transfer, the air flow, the gap of temperature inlet & outlet the underground water and the air. 3. The heat value was 6,825W when the air flow was 6,000m3/h and the gap of temperature between inlet and outlet of the underground water was 6℃, and It dropped from 25.8℃ to 23.2℃(-2.6℃ difference). The convection heat value was 88.5W/m2℃. 4. The heat value was 2.625W when the air flow was 4,000m3/h and the gap of temperature between inlet and outlet the underground water was 2℃, and It dropped from 27℃ to 22.5℃(-4.5℃ difference). The convection heat value was 33.6W/m2℃. 5. Correlation values(R2) of the testing heat values of the trial manufacture type I, II, and III were 0.9141, 0.8935, and 0.9323 respectively, and correlation values(R2) of the amount of the air flow 6,000m3/h, 5,000m3/h, 4,000m3/h were 0.9513, 0.9414, and 0.9003 respectively.

Development of heat exchanger by the utilization of underground water. I - Design for plat fin tube - (지하수 이용을 위한 열교환기 개발. I - 냉각핀의 설계제작 -)

  • Lee, W.Y.;Ahn, D.H.;Kim, S.C.;Park, W.P.;Kang, Y.G.;Kim, S.B.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.4 no.1
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    • pp.119-127
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    • 2002
  • This study was conducted to develop the heat exchanger by utilizing the heat energy of underground water(15℃), which might be used for cooling and heating system of the agricultural facilities. We developed the heat exchanger, parallel type plat fin tube made of Aluminum(Al 6063), which was named Aloo-Heat(No. of The registration design : 0247164, by Korean Intellectual property Office). The fin of exchanger was design of the granulated surface for minimizing fouling factor and dew forms, and also placed parallel to the tube in order to minimized the resistance of flows. 1. Aloo-heat was designed to have 0.03m for inside diameter, 0.036m for outside diameter of tube, 0.0012m for thickness of fin and 0.032m for length of plat fin. 2. t was also designed to have 1.5248m2/m for outside area of heat transfer, 0.0942m2/m for inside area contacting hot liquid, and the ratio (Ra) was 16.1869. 3. Efficiency of the fin was 93 percentage when fin length was 0.032m, and the fin thickness satisfied equation $\frac{h{\rho}}{k}$< 0.2 when it was 0.0012m. 4. According to the performance test of Aloo-heat, as the temperature and rate increased, the heating value also increased, heating value was 504kJ/h·m and 6,048kJ/h·m when it was 60℃, 10 𝑙/min and 80℃, 40 𝑙/min respectively. 5. The test of heating value was confident, because correlation value(R2) was 0.9898 for the temperature and 0.9721 for flow rate of hot liquid, respectively.