• Title/Summary/Keyword: 증식활성(增殖活性)

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Cloning and Expression of an $\alpha$-Amylase Gene from Bacillus circulans in B. subtilis and B. megaterium (Bacillus circulans $\alpha$-amylase 유전자의 Basillus subtilis와 Bacillus megaterium에서의 클로닝 및 발현)

  • 이동석;김지연;김한복
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.203-208
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    • 2000
  • A Baczllus circdans KCTC3004 $\alpha$-amylase gene contained in a recombinant plasmid pAL850 was transferred into a new shuttle vector plasmid pALSIlI by ligating linearlzed DNAs of pUC19 and pUB110. B. subtilis RM125 and B. megatenurn ATCC14945 transfonned with pALS111 produced the $\alpha$-amylase substantially Most of the enzyme was produced during the exponential growth period. The maxiinurn activities of the $\alpha$-amylase produced by the Bucillus transformants were compared with that of the B. circulans gene donor strain. The B. subtilis RM125(pALS111) enzyme showed the actlvicy 95 times higher than that of the gene donor cells, followed by the B, nzegaterium ATCC14945(pALSlll) enzyme with activity 34 limes higher than that of the gene donor cells. While E coli secreted about 10% of the produced enzyme, B. subtilis excreted the enzyme inlo the medium wholly and B. megaterirun about 98% ofthe total product. The plasmid pALSI11 was quite stable inB. nzegaterium (92%), inoderately stable in B. subtilis (76%), but was unstable in E. coli (38%). The SDS-PAGE and zymogram of this enzyme produced in E. coli(pALS111), B. subtilis( pALS111) or B. megateril~m (pALS111) indicated a molecular weight of 55,000. The enzymes overproduced in three different host cells hydrolyzed starch to produce mainly maltoaiose and mallooligosaccharides.

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Water Extract of Allium sativum L. Induces Apoptosis in Human Leukemia U937 Cells through Reactive Oxygen Species Generation (마늘 열수 추출물의 활성산소종 생성을 통한 인체백혈병세포의 apoptosis 유발)

  • Choi, Woo-Young;Chung, Kyung-Tae;Yoon, Tae-Kyung;Choi, Byung-Tae;Lee, Yong-Tae;Lee, Won-Ho;Ryu, Chung-Ho;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.17 no.12
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    • pp.1709-1716
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    • 2007
  • The health benefits of garlic (Allium sativum L.) are derived from a wide variety of components and from the different ways it is administered. The known health benefits of garlic include cardiovascular protective effects, stimulation of immune function, reduction of blood glucose level, protection against microbial, viral and fungal infections, as well as anticancer effects. In the present study, it was examined the effects of water extract of A. sativum (WEAS) on the growth of cultured human tumor cells in order to investigate its anti-proliferative mechanism. Treatment of WEAS to tumor cells resulted in the growth inhibition, especially in leukemia cells, which was associated with induction of G2/M arrest of the cell cycle and apoptosis. In order to further explore the critical events leading to apoptosis in WEAS-treated U937 human leukemia cells, the following effects of WEAS on components of the mitochondrial apoptotic pathway were examined: generation of reactive oxygen species (ROS), alteration of the mitochondrial membrane potential (MMP), and the expression changes of Bcl-2 and IAP family proteins. The cytotoxic effect of WEAS was mediated by its induction of apoptosis as characterized by the occurrence of DNA ladders, apoptotic bodies and chromosome condensation in U937 cells. The WEAS-induced apoptosis in U937 cells was correlated with the generation of intracellular ROS, collapse of MMP, activation of caspase-3 and down-regulation of anti-apoptotic proteins. The quenching of ROS generation with antioxidant N-acetyl-L-cysteine conferred significant protection against WEAS-elicited ROS generation, caspase-3 activation, G2/M arrest and apoptosis. In conclusion, the present study reveals that the cellular ROS generation plays a pivotal role in the initiation of WEAS-triggered apoptotic death in U937 cells.

EGCG induces Apoptosis under Hypoxic State in B16F10 Melanoma Cancer Cells (저산소증 상태에서 B16F10 피부암 세포에 EGCG를 처리하였을 때의 apoptosis 효과)

  • Kim, Yoon-Yi;Kim, In-Seop;Park, Ock-Jin;Kim, Young-Min
    • Journal of Life Science
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    • v.21 no.2
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    • pp.251-256
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    • 2011
  • EGCG, catechins in green tea, is a kind of phytochemical. Through the regulation of signal pathways, EGCG has been known to show anti-oxidant and anti-tumor effects in cells. In this study, we investigated the apoptotic effects of EGCG through AMP-activated protein kinase (AMPK) signal pathways, including hypoxia inducible factor-1 alpha (HIF-$1{\alpha}$). The experiments were performed in B16F10 melanoma cells in a hypoxic state. AMPK is activated by ATP consumption such as nutrient deficiency, exercise, heat shock, etc. The activated AMPK that plays an important role as an energy sensor inhibits proliferation of cancer cells, as well as inducing apoptosis. HIF-$1{\alpha}$, the primary transcriptional regulator of the response to oxygen deprivation, plays a critical role in modulating tumor growth and angiogenesis in a hypoxic state. The apoptotic effects of EGCG were studied in B16F10 cells in a hypoxic state. The results show that EGCG inhibits the transcriptional activity of HIF-$1{\alpha}$ and induces apoptosis. These observations suggest that EGCG may exert inhibitory effects of angiogenesis and control tumor cell growth in hypoxic melanoma cells.

Delphinidin Suppresses Angiogenesis via the Inhibition of HIF-1α and STAT3 Expressions in PC3M Cells (전립선 암세포에서 delphinidin에 의한 HIF-1α와 STAT3 억제를 통한 혈관내피 성장 인자 발현 저해 효과)

  • Kim, Mun-Hyeon;Kim, Mi-Hyun;Park, Young-Ja;Chang, Young-Chae;Park, Yoon-Yub;Song, Hyun-Ouk
    • Korean Journal of Food Science and Technology
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    • v.48 no.1
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    • pp.66-71
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    • 2016
  • Delphinidin is a blue-red pigment and one of the major anthocyanins in plants. It plays an important role in anti-oxidant, anti-inflammatory, anti-mutagenic and anti-cancer properties. In this study, we investigated the inhibitory effects of delphinidin on vascular endothelial growth factor (VEGF) gene expression, an important factor involved in angiogenesis and tumor progression in human prostate cancer. Delphinidin decreased levels of epidermal growth factor (EGF)-induced VEGF mRNA expression in PC-3M cells. The expression of the EGF-induced hypoxia inducible factor-$1{\alpha}$ (HIF-$1{\alpha}$) and signaling transducer and activator of transcription 3 (STAT3) proteins, which are the major transcription factors for VEGF, were inhibited by delphinidin. In addition, delphinidin decreases HRE-promoter reporter gene activity, suggesting that delphinidin can suppress the transcription of HIF-$1{\alpha}$ under EGF induction, leading to a decrease in the expression of VEGF. Delphinidin specifically suppressed the phosphorylation of Akt, p70S6K, and 4EBP1, but not the phosphorylation of EGFR. Therefore, our results suggest that delphinidin may inhibit human prostate cancer progression and angiogenesis by inhibiting HIF-$1{\alpha}$, STAT3 and VEGF gene expression.

Evaluation of the Cytotoxic Effects of Resveratrol Treatment with Over-the-counter Drugs on the Hepatic and Intestinal Cells (식이 폴리페놀 성분 resveratrol과 일반의약품의 복합처리에 의한 간 및 장관계 세포독성 평가)

  • Kim, Da-Ram;Kim, Mi-Ri;Hong, Jung-Il
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.217-222
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    • 2010
  • Resveratrol is a natural polyphenolic compound frequently found in grapes. The biological actions of resveratrol have been extensively investigated both in vitro and in vivo. The interactions of resveratrol with commonly-consumed drugs, however, have rarely been studied. In this study, the cytotoxic properties of resveratrol on the hepatic and intestinal cells in the presence of over-the-counter (OTC) drugs, including acetaminophen (AAP), aspirin (Asp), and ibuprofen (Ibu), were evaluated. The cytotoxic effects of resveratrol on hepatic HepG2 and colonic HCT 116 cells were not markdely changed in the presence of AAP, Asp, or Ibu. Conversely, the cytotoxicity of OTC drugs was not affected by resveratrol either. Concentrations of resveratrol below 10 mM significantly increased HepG2 cell growth after 48 or 72 hr incubation; however, the growth-stimulating effect was not observed in the presence of AAP. When HCT 116 cells were treated with OTC drugs before or after resveratrol, the cytotoxic effects were not significantly altered. The present study provides basic information for the potential health effects of the interactions between resveratrol and commonly-consumed OTC drugs.

Sex-Hormone Replacement Effect of Silkworm Pupa and Mixture with Herbs (미성숙 흰쥐에서 누에 번데기 및 한약재 혼합물의 여성호르몬 대체효과)

  • Yang Ji-Won;Choi Eun-Mi;Kwon Mu-Gil;Koo Sung-Ja
    • Korean journal of food and cookery science
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    • v.21 no.6 s.90
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    • pp.769-775
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    • 2005
  • In this study we isolate substances to serve as dietary resources in order to replace the female hormone. Silkworm (Bombyx mori) is one of the most attractive hosts for large-scale production of eukaryotic proteins which have been proven safe as a dietary resource. We report on the estrogenicity of a mixture of silkworm pupa and herbs (Ginseng,Ulkeum, and Hasuo) using the immature rat uterotrophic assay in vivo. Silkworm pupa aqueousextract (KW) and silkworm oil extract (KO) induced effects on the immature rat uterotrophic assay. KO showed neither positive uterotrophic response nor inhibition on E2 induced effect, while KW and MK (mixture of KW and herbs) showed both of the effects. It is concluded that ethanol extracts from silkworm might be a good, therapeutic, natural product for hormone-deficient diseases.

Fermentation Characteristics of Kochujang Containing Horseradish or Mustard (양고추냉이와 겨자 분말을 첨가한 고추장의 발효특성)

  • Shin, Dong-Hwa;Ahn, Eun-Young;Kim, Yong-Suk;Oh, Ji-Young
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1350-1357
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    • 2000
  • Traditional Kochujang was prepared adding horseradish or mustard powder to repress the gas formation which used to cause swelling problem during distribution. The koji for Kochujang was prepared by the strains which had high amylase and protease activities with superior flavor. The gas production from Kochujang during fermentation at $25^{\circ}C$ was ceased after stopping yeast growth completely by bactericidal components from $0.6{\sim}1.2%(w/w)$ of horseradish or mustard addition. Total viable bacterial count was not affected by adding horseradish or mustard. The amino type nitrogen content in Kochujang, which was one of the most important parameters in quality of Kochujang, increased continually during fermentation. The Kochujang fermented by P-2 isolate and added with mustard was significantly higher in amino type nitrogen content than other treatments after 120 days' fermentation. ${\alpha}-Amylase$ activity was very low while ${\beta}-amylase$ activity was high in Kochujang fermented by adding horseradish and mustard powder. The protease(acid and neutral) activities gradually increased by fermentation with no difference between treatments. The color and flavor were not different, but overall palatability of the Kochujang evaluated by sensory test showed significantly high rank in Kochujang fermented by P-2 isolate and with horseradish.

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Effects of Media and Temperatures on Micro Stem Cutting of Dendrobium nobile 'Hamana Lake Dream' × 'No. 55' in Vitro (Dendrobium nobile 'Hamana Lake Dream' × 'No.55'의 기내 줄기삽목에 미치는 배지의 종류와 온도의 영향)

  • Yoon, Jin-Young;Nam, Yu-Kyeong;Lee, Jong-Suk;Kim, Hyun-Jin
    • Journal of agriculture & life science
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    • v.44 no.3
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    • pp.23-30
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    • 2010
  • The optimal growth condition of in vitro stem cutting of Dendrobium nobile 'Hamana Lake Dream' ${\times}$ 'No. 55' was investigated. Among various media and their concentrations, MS media had better effect on the growth of micro stem cutting than Hyponex media in all concentration levels except stem length. The activated charcoal concentration in MS media showed different effects on number of stem and root, leaf length, and fresh weight: the most effective in the range of 0.1 to 1.0 g/L and barely effective above 2.0 g/L. Addition of agar 5 g/L, sucrose at 40 g/L, and peptone at 1 g/L to MS media increased significantly stem length, leaf width, and fresh weight, internode length and number of roots, and the number of stem and leaves. On the other hand, addition of gelite with any concentration had no effect on the growth of micro stem cutting compare to that of control. The optimal temperature for growth of micro stem cutting was $28^{\circ}C$. Under the same temperature, MS medium was better than Hyponex medium for the growth of stem. In addition, sucrose at 40 g/L was the most effective on growth at $28^{\circ}C$.

Neuroprotective Effects of Korean Kiwifruit against t-BHP-induced Cell Damage in PC12 Cells (국내산 참다래 추출물의 신경독성 방어효과)

  • Kim, Jeong-Hee;Yang, Hee-Kyoung;Hong, Hyun-Ju;Kang, Won-Young;Kim, Dong-Geon;Kim, Seong-Cheol;Song, Kwan-Jeong;King, Dale;Han, Chang-Hoon;Lee, Young-Jae
    • Korean Journal of Plant Resources
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    • v.23 no.2
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    • pp.165-171
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    • 2010
  • Oxidative stress induced by reactive oxygen intermediates has been implicated in a variety of human diseases including neurodegenerative disorders, cancer, cardiovascular and respiratory diseases, and mode of action of environmental toxicants. Tert-butylhydroperoxide (t-BHP) is an organic lipid hydroperoxide analogue, which is commonly used as a pro-oxidant for evaluating mechanisms involving oxidative stress in cells and tissues. In this study, the underlying mechanisms involved in the protective effects of Hwabuk 94 kiwifruit (Actinidia deliciosa cv. 'Hwabuk 94'), which is cultivated in Jeju, on the t-BHP-induced cytotoxicity in PC12 cell. The pretreatment of rat pheochromocytoma cell line PC12 with Hwabuk 94 extract ($1-100\;{\mu}g/ml$) resulted in a significant recovery from t-BHP-induced cell death and increased Bcl-2 and procaspase-3 expression, whereas the expression of Bax and cleaved PARP were decreased in a dose-dependent manner compared to the control. Furthermore, Hwabuk 94 inhibited the t-BHP-induced p38 MAP kinase and extracellular signal-regulated kinase 1/2, but not c-Jun N-terminal kinase activations. Finally, these findings suggest that Hwabuk 94 kiwifruit might attenuate t-BHP-induced PC12 cell cytotoxicity, at least in part, through the inhibition of signaling pathways mediated by the ERK1/2 and p38 MAP kinase.

Biological Properties of Propolis Isolated from Honeybees (프로폴리스의 생물학적 특성)

  • Kim, Sung-Kuk;Woo, Soon-Ok;Chang, Jong-Soo
    • Journal of Life Science
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    • v.31 no.7
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    • pp.686-697
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    • 2021
  • Propolis is a resinous substance produced by honeybees, which they use to protect their hives. Honeybees produce propolis by mixing exudates from the various trees and plants with saliva and beeswax. It has been used since around 300 B.C. as a folk medicine to cure wounds. Propolis contains many physiologically active components, such as flavonoids, phenolic compounds, and beeswax. Because of its functional components, propolis has a wide spectrum of biological applications. The compounds in propolis and its biological activity can vary according to the location of nectar source and extraction method. Propolis is most commonly known for its anti-microorganism activity against bacteria, viruses, and fungi. Artepillin C and caffeic acid phenethyl ester (CAPE) have been identified as regulatory compounds that reduce inflammation and exert immunosuppressive reactions on T lymphocytes. Through its anti-inflammatory activity, propolis exhibits anti-tumor activity, including the inhibition of cancer cell proliferation, the blocking of tumor signaling cascades, and antiangiogenesis. However, for the more apply of propolis its analysis of nectar source, identifying of propolis compound, the molecular mechanism of propolis and the investigation of compounds synergistic effects are essential. In this study, we described the physiological activity of propolis isolated from honeybees.