• Title/Summary/Keyword: galE

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High Dose of FGF-2 Induced Growth Retardation via ERK1/2 De-phosphorylation in Bone Marrow-derived Mesenchymal Stem Cells

  • Shim, Kwang Yong;Saima, Fatema Tuj;Eom, Young Woo
    • Biomedical Science Letters
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    • v.23 no.2
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    • pp.49-56
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    • 2017
  • Fibroblast growth factor (FGF)-2 is one of the most effective growth factors to increase the growth rate of mesenchymal stem cells (MSCs). Previously, we reported that low dose of FGF-2 (1 ng/ml) induced proliferation of bone marrow-derived mesenchymal stem cells (BMSCs) through AKT and ERK activation resulting in reduction of autophagy and senescence, but not at a high dose. In this study, we investigated the effects of high dose FGF-2 (10 ng/ml) on proliferation, autophagy and senescence of BMSCs for long term cultures (i.e., 2 months). FGF-2 increased the growth rate of BMSCs in a dose dependent manner for a short term (3 days), while during long term cultures (2 months), population doubling time was increased and accumulated cell number was lower than control in BMSCs when cultured with 10 ng/ml of FGF-2. 10 ng/ml of FGF-2 induced immediate de-phosphorylation of ERK1/2, expression of LC3-II, and increase of senescence associated ${\beta}$-galactosidase (SA-${\beta}$-Gal, senescence marker) expression. In conclusion, we showed that 10 ng/ml of FGF-2 was inadequate for ex vivo expansion of BMSCs because 10 ng/ml of FGF-2 induced growth retardation via ERK1/2 de-phosphorylation and induction of autophagy and senescence in BMSCs.

A Facile Synthesis of Ethyl 2,6-Di-O-benzyl-3-O-(3,4,6-tri-O-acetyl-2-deoxy-2-N-phthalimido-$\beta$-D-glucopyranosyl)-1-thio-$\beta$-D-galactopyranoside and Studies on the Regioselectivity of 2,3,4-OH Groups of Galactosyl Acceptor (Ethyl 2,6-Di-O-benzyl-3-O-(3,4,6-tri-O-acetyl-2-deoxy-2-N-phthalimido-$\beta$-D-glucopyranosyl)-1-thio-$\beta$-D-galactopyranoside의 합성법의 개발 및 갈락토스 받게의 2,3,4-OH기들의 위치 선택성에 대한 연구)

  • Park, Jung Soo;Yoon, Shin Sook;Yun, Mi Kyung;Chun, Keun Ho;Nam Shin, Jeong E.
    • Journal of the Korean Chemical Society
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    • v.42 no.5
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    • pp.549-558
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    • 1998
  • O-Antigenic part of Campylobacter jejuni gram negative bacteria was reported to consist of a repeated trisaccharide unit. The disaccharides, GlcNAc-Gal derivatives, as key intermediates for the synthesis of trisaccharide repeating units were synthesized. At the $\beta(1{\to}3)$ glycoside bond formation step between 3,4,6-tri-O-acctyl-2-deoxy-2-N-phthalimido-${\beta}$-D-glucopyranosyl bromide and galactosyl acceptors, high regioselectivities between 2, 3, and 4-OH groups of galactosyl acceptors were found. As a result, no further selective protection steps for OH groups of galactosyl acceptors was necessary, and more effective and compact synthetic scheme was achieved.

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Construction of New Transfer Vector of Nuclear Polyhedrosis Virus of the Silkworm, Bombyx mori (누에 핵다각체병 바이러스를 이용한 새로운 전이 벡터의 제작)

  • 우수동;김우진;진병래;강석권
    • Journal of Sericultural and Entomological Science
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    • v.37 no.1
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    • pp.46-51
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    • 1995
  • In order to develope baculovirus expression vector system, we constructed new transfer vector of nuclear polyhedrosis virus of the silkworm, Bombyx mori. The promoter region containing only adenine of translation start codon of polyhedrin gene was cloned by polymerase chain reaction technique. And the 5' and 3' leader regions of polyhedrin gene was sequentially cloned. The polyhedrin coding gene was deleted from the +2 to the +597 position. As the result, we constructed new transfer vector which has EcoRI, SacI and KpnI sites for the cloning sites of foreign gene. New transfer vector was named as pBmKSKl. Escherichia coli $\beta$-galactosidase gene as foreign gene was inserted into pBmKSKl, under the control of the polyhedrin promoter and expressed in B. mori cells. The result showed that the new transfer vector pBmKSK1 is functional.

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Alteration of Innate Immune T and B Cells in the NC/Nga Mouse (아토피성 피부질환 동물 모델 NC/Nga 생쥐에서 내재면역 T와 B 세포의 변형)

  • Kim, Jung-Eun;Kim, Hyo-Jeong;Kim, Tae-Yoon;Park, Se-Ho;Hong, Seok-Mann
    • IMMUNE NETWORK
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    • v.5 no.3
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    • pp.137-143
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    • 2005
  • Background: Millions of people in the world are suffering from atopic dermatitis (AD), which is a chronic inflammatory skin disease triggered by Th2 immune responses. The NC/Nga mouse is the most extensively studied animal model of AD. Like human AD, NC/Nga mice demonstrate increased levels of IgE, a hallmark of Th2 immune responses. Adaptive immunity cannot be generated without help of innate immunity. Especially natural killer T (NKT) cells and marginal zone B (MZB) cells have been known to play important roles in linking innate immunity to adaptive immunity. Methods: Through flow cytometric analysis and ELISA assay, we investigated whether these lymphocytes might be altered in number in NC/Nga mice. Results: Our data demonstrated that the number of NKT cells was reduced in NC/Nga mice and IFN${\gamma}$ production by NKT cells upon ${\alpha}-GalCer$ stimulation decreased to the levels of CD1d KO mice lacking in NKT cells. However, reduction of NKT cells in NC/Nga mice was not due to CD1d expression, which was normal in the thymus. Interestingly, there was a significant increase of $CD1d^{high}B220^+$ cells in the spleen of NC/Nga mice. Further, we confirmed that $CD1d^{high}B220^+$ cells are B cells, not dendritic cells. These $CD1d^{high}B220^+$ B cells show $IgM^{high}CD21^{high}CD23^{low}$, a characteristic phenotype of MZB cells. Conclusion: We provide the evidence that there are decreased activities of NKT cells and increased number of MZB cells in the NC/Nga mice. Our findings may thus explain why NC/Nga mice are susceptible to AD.

Glucosylation of Isoflavonoids in Engineered Escherichia coli

  • Pandey, Ramesh Prasad;Parajuli, Prakash;Koirala, Niranjan;Lee, Joo Ho;Park, Yong Il;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.37 no.2
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    • pp.172-177
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    • 2014
  • A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-${\alpha}$-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4' and 7 hydroxyl groups, but not at the $5^{th}$ hydroxyl group of the A-ring, resulting in the production of genistein 4'-O-${\beta}$-D-glucoside, genistein 7-O-${\beta}$-D-glucoside (genistin), genistein 4',7-O-${\beta}$-D-diglucoside, biochanin A-7-O-${\beta}$-D-glucoside (sissotrin), daidzein 4'-O-${\beta}$-D-glucoside, daidzein 7-O-${\beta}$-D-glucoside (daidzin), daidzein 4', 7-O-${\beta}$-D-diglucoside, and formononetin 7-O-${\beta}$-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOF-ESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli $BL21(DE3)/{\Delta}pgi{\Delta}zwf{\Delta}ushA$ overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of $200{\mu}M$ of supplemented isoflavonoids, without any additional UDP-${\alpha}$-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides.

Functional Chemical Components in Protaetia brevitarsis Larvae: Impact of Supplementary Feeds

  • Yoon, Chul-Ho;Jeon, Seung-Ho;Ha, Yeon Jo;Kim, Sam Woong;Bang, Woo Young;Bang, Kyu Ho;Gal, Sang Wan;Kim, Il-Suk;Cho, Young-Son
    • Food Science of Animal Resources
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    • v.40 no.3
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    • pp.461-473
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    • 2020
  • The goal of this study was to evaluate the influence of various supplementary feeds on the chemical composition and production of bioactive substances in Protaetia brevitarsis larvae. The primary feed-oak-fermented sawdust-was supplemented with a variety of substances, including aloe, apple, banana, sweet persimmon (S. persimmon) and sweet pumpkin (S. pumpkin). Crude protein and fat content were the highest in the control and S. pumpkin group, respectively. Supplementary feeds increased the content of unsaturated fatty acids, except in the group receiving S. pumpkin, in which oleic acid was the most abundant (58.2%-64.5%). Free essential amino acids in larvae receiving supplementary aloe were higher compared with the control group except for Lys and His. Polyphenol and flavonoid contents and the antioxidant activities of ABTS and DPPH were higher in all treated groups compared with the control group. Although supplementary feeds led to a decreased crude protein content in the treated larvae when compared with the control group, these treatments generally improved the levels of unsaturated fatty acids and antioxidative activity. Therefore, we suggest that among the supplementary foods tested, aloe is a better resource for P. brevitarsis based on crude protein content, free amino acids and other bioactive compounds such as unsaturated fatty acids and antioxidants.

비단가리비 인공종묘 생산

  • 박기열
    • Korean Aquaculture
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    • v.14 no.2
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    • pp.52-65
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    • 2002
  • 비단가리비의 인공 종묘 생산을 목적으로 전라남도 신안군 대흑산도 주변에 서식하는 비단가리비를 대상으로 인공 종묘 생산을 위한 산란 유발, 수정란의 발생 과정, 유생 사육, 채묘 및 중간 육성 등 양식 생물학적 연구를 실시하였다. 어미의 각종 산란 유발 자극에 대한 반응은 Serotonin 주사, 온도 자극, 혼합 자극에서 반응율이 가장 높았으며, 자외선 조사 해수 자극은 수컷만 반응을 하였고 간출 자극은 반응이 없어 실효성이 없는 것으로 나타났다. 수정란의 크기는 $69.5{\mu}m$이었고, 수정 후 약 2시간에 2 세포기, 8시간 후에 8 세포기, 20시간 후에는 담륜자 유생으로 부화하였으며 40시간 후에는 D상 유생으로 되었다. 수온별 비단가리비 유생의 성장은 수온 $20^{\circ}C$에서 각장 $178.9{\mu}m$로 가장 좋았으며, 이 때의 생존율은 15.5%이었다. 그러나 수온 $15^{\circ}C$에서는 낮은 성장을 보여 각장 $135.9{\mu}m$로 성장하였으며, 생존율도 9.8%로 저조하였다. 수용 밀도별 사육 시험에서 $1m{\ell}$당 1개체와 5개체에서 성장 및 생존율이 양호하여 성장 및 생존율을 볼 때 적정 사육 밀도는 $1m{\ell}$당 5개체 이하로 나타났다. 먹이 생물 종류에 따른 유생의 성장을 알기 위하여 I, gal-baba, C, calcitrans, N, oculata를 단독 또는 혼합으로 공급하였을 때 실험 종료후 각장의 성장은 I, galbana+C. calcitrans+N. oculata구가 $194.2{\mu}$로 가장 좋았고 N. oculata구는 $162.2{\mu}m$로 가장 낮은 성장을 보였다. 생존율에서는 I. galbana+C. calcitransoculata구는 9.4%로 가장 낮은 생존율을 보였다. 채묘기별 유생의 부착률은 염화비닐판을 수평으로 놓은 것과 패각이 각각 3.43%와 3.17%로 가장 양호하였으나, 양파망과 염화비닐판을 수직으로 놓은 것은 각각 1.52%와 1.61%로 비교적 저조하였다. 수정 후 40일째부터 90일째까지 측정한 부착치패의 경과 일수에 따른 각장의 성장은 $SL=184.44e^{0.0335X}(r^2=0.9861)$의 회귀직선식으로서 나타났다. 중간육성 시험에서 수심별 성장을 분석한 결과, 비단가리비 치패는 저층보다 표층이 각장 5.92mm, 전중량 6.07g 정도 더 빨리 성장하였다.

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Study of Anti-Alzheimer Activities from Scrophularia buergeriana Water Extract by Alzheimer's Protein APP-transgenic Fly (현삼(玄蔘) 수추출물(水抽出物)이 아밀로이드 전구단백질(前驅蛋白質)로 형질전환(形質轉換)된 초파리에 미치는 효과)

  • Kim, Jin-Woo;Lee, Soon-E;Lee, Jong-Hwa;Min, Sang-Jun;Kim, Tae-Heon;Lyu, Yeoung-Su;Kang, Hyung-Won
    • Journal of Oriental Neuropsychiatry
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    • v.20 no.2
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    • pp.121-131
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    • 2009
  • Objectives : From Scrophularia buergeriana water extract(SBW), has been used in vivo test for its beneficial effects on neuronal survival and neuroprotective functions, particularly in connection with APP-related dementias and Alzheimer's disease(AD). $A{\beta}$ oligomer derived from proteolytic processing of the ${\beta}$-amyloid precursor protein(APP), including the amyloid-${\beta}$ peptide($A{\beta}$), play a critical role in the pathogenesis of Alzheimer's dementia. Methods : Using drosophila APP model on APP-induced neuronal cytotoxicity, we demonstrated that SBW prevents neurotoxicity of $A{\beta}$ oligomer, which are the behavior, and possibly causative, feature of AD. We investigated the neuroprotective effects of SBW against the effects of oligomeric $A{\beta}$ and fly behaveior and life span by UAS-GRIM/APP-GAL within transgenic flies. Results and Conclusions : SBW repaired damage leading to the behaveior of APP-induced fly and delayed life span. These results suggest that neuronal damage in AD might be due to two factors: a direct $A{\beta}$ oligomer toxicity and multiple cellular and molecular neuroprotective mechanisms, including attenuation of apoptosis and direct inhibition of $A{\beta}$ oligomer, underlie the neuroprotective effects of SBW.

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Tocopherols and fatty acids in plant seeds from Korea (한국산 두류 종실중 토코페롤 함량 및 지방산 조성)

  • Lee, In-Bog;Choi, Kang-Ju;Yu, Kwang-Keun;Chang, Ki-Woon
    • Applied Biological Chemistry
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    • v.35 no.1
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    • pp.1-5
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    • 1992
  • The concentrations of tocopherols and lipids, and compositions of fatty acids in the seeds of thirteen pulse plants, which are used for the raw materials of our traditional food as well as for the typical nutrients in Korea, were analyzed by HPLC and GC. The beans showed great differences of the tocopherol concentrations from 4.74 to 44.05 mg/100g, which were in the order of ${\gamma}->{\delta}->{\alpha}-tocopherol$ with the exception of the order of ${\delta}->{\gamma}->{\alpha}-tocopherol$ in jeock sodu and gal sodu of azuki bean. The contents of lipids were also remarkably different as 0.53 to 20.96%. Especially, the beans having from 15.78 to 20.96% of lipid contents showed high composition ratios of unsaturated fatty acid between 85.6 to 88.2%, of which the contents of linoleic acid were the highest as 56.0 to 58.6% of the total fatty acids, and also showed comparatively high concentrations of tocopherols as 23.2 to 44.05 mg/100g. There were significantly positive correlation between ${\gamma}-tocopherol$ and unsaturated fatty acids.

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Studies on Microbial Extracellular $\beta$-Gala-ctosidase

  • Lee, Keun-Eok
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1979.04a
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    • pp.113.2-114
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    • 1979
  • $\beta-Galactosidase$ is an enzyme which catalizes hydrolysis of lactose, a natural substrate, to glucose and galctose and transferring some monosac-charide units to active acceptors as sugar or alcohol. The occurence of $\beta-Galactosidase$ is known in various microorganisms, animals and higher plants and has been studied by many investigatigators. Especially, a great deal of articles for the enzyme of E. coli have been presented in genetic control mechanism and induction-repression effects of proteins, On the other hand, in the dairly products industry, it is important to hydrolyes lactosd which is the principal sugar of milk and milk products. During the last few years, the interest in enzymatic hydrolysis of milk lactose has teen increased, because of the lactose intolerence in large groups of the population. Microbial $\beta-Galactosidases$ are considered potentially most suitable for processing milk to hydrolyse lactose and, in recent years, the immobilized enzyme from yeast has been examined. Howev, most of the microbial $\beta-Gal$ actosidase are intracellular enzymes, except a few fungal $\beta-Gala-$ ctosidases, and extracellular $\beta-Galactosidase$ which may be favorable to industrial applieation is not so well investigated. On this studies, a mold producing a potent extracellular $\beta-Galactosidase$ was isolated from soil and identified as an imperfect fungus, Beauveria bassians. In this strain, both extracellular and intracellular $\beta-Galactosidases$ were produced simultaneously and a great increase of the extracellular production was acheved by improving the cultural conditions. The extracellular enzyme was purified more than 1, 000 times by procedures including Phosphocellulose and Sephadex G-200 chromatographies. Several characteristics of the enzymewas clarified with this preparation. The enzyme has a main subunit of molecular weight of 80, 000 which makes an active aggregate. And at neutral pH range, it has optimum pH for activity and stability. The Km value was determined to be 0.45$\times$10$^{-3}$ M for $o-Nitrophenyl-\beta-Galactoside.$ In any event, it is interesting to sttudy the $\beta-Galactosidase$ of B. bassiana for the mechanism of secretion and conformational structure of enzyme.

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