• Title/Summary/Keyword: Gal1

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The Effect of Ginsenosides on Galactosamine-induced Hepatotoxicity (인삼 사포닌이 간세포 독성에 미치는 영향)

  • kim, Sun-Yeou;Kim, Young-Choong;Byun, Soon-Jung;Kim, Eun
    • Korean Journal of Pharmacognosy
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    • v.22 no.4
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    • pp.219-224
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    • 1991
  • Liver protective effects of ginsenosides as well as fractions of dammarane glycosides of Panax ginseng were studied using galactosamine (GalN)-induced cytotoxicity in primary cultured rat hepatocytes. Preventing effects on GalN-induced hepatotoxicity were found both microscopic observation and determination of GPT level with total dammarane glycosides fraction and $20(S)-ginsenoside-Rb_1$ as well as $20(S)-ginsenoside-Rg_1$ at the concentration of $50{\mu}g/ml$. The syntheses of both protein and RNA were significantly increased by the treatment of $50{\mu}g/ml$ of total dammarane glycoside fraction, $20(S)-ginsenoside-Rb_1$, -Rc, -Re and $-Rg_1$, respectively in both normal and GalN-induced cytotoxic hepatocytes.

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Motional Properties in the Structure of GlcNAc(β1,3)Gal(β)OMe Studied by NMR Spectroscopy and Molecular Modeling

  • 심규창;이상원;김양미
    • Bulletin of the Korean Chemical Society
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    • v.18 no.4
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    • pp.415-424
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    • 1997
  • Conformational flexibilities of the GlcNAc(β1,3)Gal(β)OMe are investigated through NMR spectroscopy and molecular modeling. Adiabatic energy map generated with a dielectric constant of 50 contains three local minima. All of the molecular dynamics simulations on three local minimum energy structures show fluctuations between two low energy structures, N2 at φ=80° and ψ=60° and N3 at φ=60° and ψ=-40°. We have presented adequate evidences to state that GlcNAc(β1,3)Gal(β)OMe exists in two conformationally discrete forms. Two state model of N2 and N3 conformers with a population ratio of 40:60 is used to calculate the effective cross relaxation rate and reproduces the experimental NOEs very well. Molecular dynamics simulation in conjunction with two state model proves successfully the dynamic equilibrium existed in GlcNAc(β1,3)Gal(β)OMe and can be considered as a powerful method to analyze the motional properties in the structure of carbohydrate. This observation also cautions against the indiscriminate use of a rigid model to analyze NMR data.

Generation of Female Porcine Fibroblasts Expressing Efficiently Membrane Cofactor Protein at ${\alpha}1$,3-Galactosyltransferase locus (${\alpha}1$,3-Galactosyltransferase 유전자 좌위에서 Membrane Cofactor Protein을 효과적으로 발현하는 자성 돼지 섬유아세포의 생산)

  • Oh, Keon Bong;Kim, Bella;Hwang, Seongsoo;Ock, Sun-A;Im, Seoki;Park, Jin-Ki
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.289-295
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    • 2013
  • Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of ${\alpha}1$,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human $EF1{\alpha}$ promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human $EF1{\alpha}$ promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.

Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

Characterization of Bioactive Sialyl Oligosaccharides Separated from Colostrum of Indonesia Dairy Goat

  • Taufik, Epi;Arief, Irma Isnafia;Budiman, Cahyo;Wibisono, Yusuf;Noviyanto, Alfian
    • Food Science of Animal Resources
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    • v.42 no.3
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    • pp.426-440
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    • 2022
  • The bioactive functions of oligosaccharides from human milk have been reported by many studies. Many of oligosaccharides isolated from colostrum and/or milk of dairy animals have been reported to have similar chemical structures with those in human colostrum and/or milk. It has been proved by several studies that the oligosaccharides with similar chemical structure shared common bioactivities. Among domesticated dairy animals, bovine/cattle, caprine/goat, and ovine/sheep are the most commonly used species to isolate oligosaccharides from their colostrum and/or milk. Several studies on the oligosaccharides from goat colostrum and milk have revealed similar properties to that of human milk and possess the highest content of sialyl oligosaccharides (SOS) as compared to other ruminants. Indonesia ranks first in Association of Southeast Asian Nations (ASEAN) for goat milk production. Therefore, goat milk is the second most consumed milk in the country. The most reared dairy goat breed in Indonesia is Etawah Grade. However, oligosaccharides from Indonesia dairy animals including goat, have not been characterized. This is the first study to characterize oligosaccharides from Indonesia dairy animals. The present study was aimed to isolate and characterize oligosaccharides, specifically SOS from the colostrum of Etawah Grade goats by using proton/1H-nuclear magnetic resonance. The SOS successfully characterized in this study were: Neu5Ac(α2-3)Gal(β1-4)Glc (3'-N-acetylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4)Glc (6'-N-acetylneuraminyllactose), Neu5Gc(α2-3)Gal(β1-4)Glc (3'-N-glycolylneuraminyllactose), Neu5Gc(α2-6)Gal(β1-4)Glc (6'-N-glycolylneuraminyllactose), Neu5Ac(α2-6)Gal(β1-4) GlcNAc (6'-N-acetylneuraminyllactosamine) and Neu5Gc(α2-6)Gal(β1-4)GlcNAc (6'-N-glycolylneuraminyllactosamine). This finding shows that Etawah Grade, as a local dairy goat breed in Indonesia, is having significant potential to be natural source of oligosaccharides that can be utilized in the future food and pharmaceutical industries.

The Effect of Differences between Gal-Ot and Undyed Clothing and clothing Types on Wear Sensation (갈옷과 흰옷 및 그 의복형태의 차가 착용감에 미치는 영향)

  • 박순자;손원교
    • Journal of the Korean Society of Clothing and Textiles
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    • v.23 no.1
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    • pp.30-41
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    • 1999
  • This study was conducted to verify the advantages of Gal-Ot. Gal-Ot is defined the clothings dyed with persimmon juice. Firstly the physical properties of fabrics were examined. Secondly the wear tests were conducted. The wear tests were performed in climatic chamber controlled 3$0^{\circ}C$ air temperature 50$\pm$5% R.H and 0.2m/s air movement. Four women subjects participated in this experiment and 4 times experiments were performed per one subject. The expeimental schedule was planned following 4 steps that is sedentary posture during 30 minutes-walking the 5。slope treadmill by 70m/min during 20 minutes-sedentary posture during 20 minutes-standing posture on toward the blowing wind during 10 minutes. The results obtained is as folows : The insulation of fabric was increased with dyeing with persimmon juice. The air permeability of fabric was remarkably increased with dyeing with persimmon juice. The mean skin temperatures of subjects were apt to be higher in wearing Gal-Ot than undyed clothing. The clothing micro temperature of subjects were slightly inclined to become lower in wearing Gal-Ot than undyed clothing, The clothing micro humidity was decreasee when: the subjects were gal-Ot than wore undyed clothing, And then it brought about more comfortable on wear sensation. The differences of physiological reactions were not consistent between clothing types except for sweat rate on the back. This result may be attributed the to complex experimental schdules consisted of four steps and to a little difference between open type and close type. However I consider that the clothing type of Gal-Ot is suitable for open type because Gal-Ot is summer wear for blocking the sultry heat.

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Transdifferentiation of α-1,3-galactosyltransferase knockout pig bone marrow derived mesenchymal stem cells into pancreatic β-like cells by microenvironment modulation

  • Ullah, Imran;Lee, Ran;Oh, Keon Bong;Hwang, Seongsoo;Kim, Youngim;Hur, Tai-Young;Ock, Sun A
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.11
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    • pp.1837-1847
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    • 2020
  • Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.

Preparation of $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ by Bacillus sp. ${\beta}-mannanase$ and Growth Activity to Intestinal Bacteria (Bacillus sp.유래 ${\beta}-mannanase$에 의한 $Gal^3Man_4(6^3-mono-{\alpha}-D-galacto-pyranosyl-{\beta}-mannotetraose)$ 조제 및 장내세균에 대한 생육활성)

  • Kim, Sang-Woo;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.379-383
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    • 2004
  • For the elucidation of substrate specificity to the brown copra meal by Bacillus sp. ${\beta}-mannanase.$, the enzymatic hydrolysate after 24 hr of reaction was heated in a boiling water bath for 10 min, and then centrifuged to remove the insoluble materials from hydrolysates. The major hydrolysates composed of D.P 5 and 7 galactosyl mannooligosaccharides. For the separate of galactosyl mannooligosaccharides, the supernatant solution of 150 ml was put on a first activated carbon column. The column was then washed with 5 l of water to remove mannose and salts. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol, at the flow rate of 250 ml per hour. The sugar composition in each fraction tubes was examined by TLC and FACE analysis. The combined fraction from F3 was concentrated to 30 ml by vacuum evaporator. Then put on a second activated carbon column. The oligosaccharides in the column were eluted by a liner gradient of $0{\sim}30%$ ethanol (total volume: 5 l), at the flow rate of 250 ml per hour. The eluent was collected in 8 ml fraction tubes, and the total sugar concentration was measured by method of phenol-sulfuric acid. The major component of F2 separated by 2nd activated carbon column chromatography were identified $Gal^3Man_4(6^3-mono-{\alpha}-D-galactopyranosyl-{\beta}-mannotetraose)$. To investigate the effects of brown copra meal galactomannooligosaccharides on growth of Bifidobacterium longum, B. bifidum were cultivated individually on the modified-MRS medium containing carbon source such as $Gal^3Man_4$, compared to those of standard MRS medium.

Determination of the Gravity Anomaly in the Ocean Area of Korean Peninsula using Satellite Altimeter Data (위성 고도자료를 이용한 한반도 해상지역에서의 중력이상의 결정)

  • 김광배;최재화;윤홍식;이석배
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.13 no.2
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    • pp.177-185
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    • 1995
  • Gravity anomalies were recovered on a $5'\times{5'}$grid using the sea surface height data obtained from the combination of Geosat, ERS-1, Topex/Poseidon altimeter data around Korean Peninsula bounded by latitude between $30^\circ{N}\;and\;50^\circ{N}$ and longitude $120^\circ{E}\;to\;140^\circ{E.}$ In order to recover the gravity anomalies from SSH(Sea Surface Height), inverse FFT technique was applied. The estimated gravity anomalies were compared with gravity anomalies measured by shipboard around Korean Peninsula. In comparison with the differences of gravity anomaly between measured data and altimeter data, the mean and the standard deviation were found to be -0.51 mGal and 13.48 mGal, respectively. In case of comparison between the measured data and the OSU91A geopotential model, the mean and the standard deviation were found to be 11.93 mGal and 19.19 mGal, respectively. The comparison of gravity anomalies obtained from the OSU91A geopotential model and the altimeter data was carried out. The results were mean of 5.30 meal and standard deviation of 19.62 mGal. From the results, we could be concluded that the gravity anomalies computed from the altimeter data is used to the geoid computation instead of the measured data.

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Hepatoprotective Activity of Dandelion (Taraxacum officinale) Water Extract against D-Galactosamine-Induced Hepatitis in Rats (D-Galactosamine으로 유발된 간손상에 대한 민들레 열수추출물의 예방효과)

  • Park, Ji-Young;Park, Chung-Mu;Kim, Jin-Ju;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.177-183
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    • 2008
  • This study aimed to investigate the protective effect of dandelion water extract (DWE) on liver injury induced by D-galactosamine (GalN) in Sprague-Dawley rats. Fifty rats were divided into 5 groups; normal control (C), DWE-control (DWE-C: saline injection after feeding 3% DWE diet), GalN-control (GalN-C: GalN injection after normal diet), DWE I (GalN injection after feeding 1.5% DWE diet), and DWE II (GalN injection after feeding 3% DWE diet). After 2 weeks, the acute hepatitis was induced by GalN (650 mg/kg, i.p.) and 24 hrs later, all rats were sacrificed. The DWE supplement ameliorated the serum alanine and aspartate aminotransferase (AST, ALT) as well as alkaline phosphatase (ALP) and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$. Hepatic antioxidative enzyme activities, such as catalase, GSH peroxidase, GSH reductase, and Mn-superoxide dismutase (SOD) were slightly or significantly elevated by the treatment of DWE. Moreover, the histological examination corresponded with these biochemical observations. According to these findings, dandelion could be used as a potential therapeutic material for treating chemically induced acute hepatitis.