• Title/Summary/Keyword: Gal1

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Ginsenoside Rg1 supplementation clears senescence-associated β-galactosidase in exercising human skeletal muscle

  • Wu, Jinfu;Saovieng, Suchada;Cheng, I-Shiung;Liu, Tiemin;Hong, Shangyu;Lin, Chang-Yu;Su, I-Chen;Huang, Chih-Yang;Kuo, Chia-Hua
    • Journal of Ginseng Research
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    • v.43 no.4
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    • pp.580-588
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    • 2019
  • Background: Ginsenoside Rg1 has been shown to clear senescence-associated beta-galactosidase (SA-${\beta}$-gal) in cultured cells. It remains unknown whether Rg1 can influence SA-${\beta}$-gal in exercising human skeletal muscle. Methods: To examine SA-${\beta}$-gal change, 12 young men (age $21{\pm}0.2years$) were enrolled in a randomized double-blind placebo controlled crossover study, under two occasions: placebo (PLA) and Rg1 (5 mg) supplementations 1 h prior to a high-intensity cycling (70% $VO_{2max}$). Muscle samples were collected by multiple biopsies before and after cycling exercise (0 h and 3 h). To avoid potential effect of muscle biopsy on performance assessment, cycling time to exhaustion test (80% $VO_{2max}$) was conducted on another 12 participants (age $23{\pm}0.5years$) with the same experimental design. Results: No changes of SA-${\beta}$-gal were observed after cycling in the PLA trial. On the contrary, nine of the 12 participants showed complete elimination of SA-${\beta}$-gal in exercised muscle after cycling in the Rg1 trial (p < 0.05). Increases in apoptotic DNA fragmentation (PLA: +87% vs. Rg1: +133%, p < 0.05) and $CD68^+$ (PLA:+78% vs. Rg1:+121%, p = 0.17) occurred immediately after cycling in both trials. During the 3-h recovery, reverses in apoptotic nuclei content (PLA:+5% vs. Rg1 -32%, p < 0.01) and increases in inducible nitrate oxide synthase and interleukin 6 mRNA levels of exercised muscle were observed only in the Rg1 trial (p < 0.01). Conclusion: Rg1 supplementation effectively eliminates senescent cells in exercising human skeletal muscle and improves high-intensity endurance performance.

Computation of Complete Bouguer Anomalies in East Sea (동해 지역의 완전부우게 이상 계산)

  • Kim, Young-Hyun;Yun, Hong-Sik;Lee, Dong-Ha;Huang, He
    • Proceedings of the Korean Society of Surveying, Geodesy, Photogrammetry, and Cartography Conference
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    • 2010.04a
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    • pp.165-168
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    • 2010
  • This paper describes the results of complete Bouguer anomalies computed from the Free-air anomalies that derived from Sandwell and DNSC08 mairne gravity models. Complete bouguer corrections consist of three parts: the bouguer correction (Bullard A), the curvature correction (Bullard B) and the terrain correction (Bullard C). These all corrections have been computed over the East Sea on a $1'{\times}1'$ elevation data (topography and bathymetry) derived from ETOPO1 global relief model. In addition, a constant topographic (sea-water) density of $2,670kg/m^3$ ($1,030kg/m^3$) has been used for all correction terms. The distribution of complete bouguer anomalies computed from DNSC08 are -34.390 ~ 267.925 mGal, and those from Sandwell are -32.446 ~ 266.967 mGal in East Sea. The mean and RMSE value of the difference between DNSC08 and Sandwell is $0.036{\pm}2.373$ mGal. The highest value of complete bouguer anomaly are found around the region of $42{\sim}43^{\circ}N$ and $137{\sim}139^{\circ}E$ (has the lowest bathymetry) in both models. Theses values show that the gravity distribution of both models, DNSC08 and Sandwell, are very similar. They indicate that satellite-based marine gravity model can be effectively used to analyze the geophysical, geological and geodetic characteristics in East Sea.

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Construction of Recombinant Bombyx mori Nuclear Polyhedrosis Virus Using a FLP/FRT System of Yeast, Saccharomyces cerevisiae 2$\mu$m plasmid (Yeast의 FLP/FRT 시스템을 이용한 BmNPV의 유전자 재조합)

  • 강석우;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.40 no.1
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    • pp.52-59
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    • 1998
  • For the construction of plasmid and bmNPV sarrying the FRT recognition site for the FLP recombinases, we synthesized the wild type FRT dligonucleotides. The target FRT sequences consist of three 13bp repeated DNA sequences; two repeats in a direct orientation and one inverted relative to the other two. In addition, there is an 8bp spacer region between the repeats which determune the orientation of the FRT recombination site. In order to place the FRT site both in target BmNPV genome and the transfer vector, we constructed a plasmid, FRT site both in the target BmNPv genome and the transfer vector, we constructed a plasmid, pFRT$\beta$-gal, carrying the FRT sites within the cloning sites of pSV vector and a recombinant BmNPV, vFRTPH, carrying the FRT sites at a downstream of polyhedrin promotor, respectively. In order to test the functionality of the FLP/FRT site-specific recombination system, vFRTPH, pFRT$\beta$-gal and pHsFLP DNA were co-transfected into BmN-4 cells. The resulting recombinant virus was designated a vFRT$\beta$2-gal. From construction analysis of the vFRT$\beta$2-gal with PCR technique it was concluded that the entire pFRT$\beta$-gal plasmid with $\beta$-galactosidase gene and origines of replication flanked by two functional hybrid FRT sequences. The efficiency of recombination was 8.7%, which was higher than that(2.2%) of recombination between a conventional transfer vector and the wild type BmNPV.

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Effect of Medium Components on the Productivity of Fibrinolytic Enzyme in Bacillus sp (배지 조성에 따른 Bacillus sp. 의 혈전 용해효소 생산효과)

  • 김영숙
    • Journal of Life Science
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    • v.9 no.4
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    • pp.489-492
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    • 1999
  • A bacteial strain which can produce the extracellular fibrinolytic enzyme was isolated from Jeot-Gal (anchovy) that was Korean traditional salt-fermented fish. The isolated bacterium was identified to be a strain of Bacillus sp. The optimal medium for fibrinolytic enzyme production was determined to consist of 5 g maltose, 10 g defatted soybean, 20 g sodium chloride, 1.75 g K2HPO4 per liter (pH 7.0)

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Characterization of the $\alpha$-Galactosidase Gene from Leuconostoc mesenteroides SY1

  • KIM JONG HWAN;PARK JAE-YONG;JEONG SEON-JU;CHUN JIYEON;LEE JONG HOON;CHUNGZ DAE KYUN;KIM JEONG HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.800-808
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    • 2005
  • Leuconostoc mesenteroides SY1, an isolate from kimchi, was able to ferment $\alpha$-galactosides, such as melibiose and raffinose. $\alpha$-Galactosidase ($\alpha$-Gal) activity was higher in cells grown on melibiose and raffinose than cells grown on galactose, sucrose, and fructose. $\alpha$-Gal activity was not detected in cells grown on glucose, indicating the operation of carbon catabolite repression (CCR). A 6 kb DNA fragment was PCR amplified using a primer set based on the nucleotide sequence of a putative $\alpha$-galactosidase gene (aga) from L. mesenteroides ATCC 8293. Nucleotide sequencing of the 6 kb fragment confirmed the presence of aga and other genes involved in the galactosides utilization, and the gene order was galR (transcriptional regulator)-aga-gaIK (galactokinase)-gaIT (galactose-1-phosphate uridylyltransferase). Northern blotting experiment showed that aga, gaIK, and gaIT constituted the same operon, that the transcription was induced by galactosides, such as melibiose and raffinose, whereas gaIR was independently transcribed as a monocistronic gene, and that the level of transcription was fairly constant. The aga was overexpressed in E. coli BL21 (DE3) using pET26b(+) vector, and $\alpha$-Gal was accumulated in E. coli as an inclusion body.

Protective effect of ultrasonication-processed ginseng berry extract on the D-galactosamine/lipopolysaccharide-induced liver injury model in rats

  • Nam, Yoonjin;Bae, Jinhyung;Jeong, Ji Hoon;Ko, Sung Kwon;Sohn, Uy Dong
    • Journal of Ginseng Research
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    • v.42 no.4
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    • pp.540-548
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    • 2018
  • Background: Acute hepatic failure is a life-threatening critical condition associated with rapid deterioration of liver function and liver transplantation. Several studies have shown that Panax ginseng Mayer has antidiabetic and hepatoprotective effects. However, the hepatoprotective effect of ginseng berry is still unveiled. In this study, we evaluated the hepatoprotective effects of ultrasonication-processed ginseng berry extract (UGBE) on acute hepatic failure model in rats. Methods: Ginseng berry extract (GBE) was ultrasonically processed. The GBE, silymarin, and UGBE were orally administered to male Sprague-Dawley rats for 4 wk. Twenty-four h after the last administration, rats were challenged with D-galactosamine (D-GalN)/lipopolysaccharide (LPS). Results: After ultrasonication, the component ratio of ginsenosides Rg2, Rg3, Rh1, Rh4, Rk1, Rk3, and F4 in GBE had been elevated. Administration of UGBE significantly increased the survival rate of D-GalN/LPS-challenged rats. Pretreatment with UGBE significantly decreased serum alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in D-GalN/LPS-challenged rats in a dose-dependent manner. The levels of enzymatic markers for oxidative stress (superoxide dismutase, glutathione peroxidase, catalase, and glutathione) were increased by UGBE treatment in a dose-dependent manner. Tumor necrosis factor alphalevel, inducible nitric oxide synthase activities, and nitric oxide productions were reduced by UGBE treatment. In addition, hemeoxygenase-1 levels in liver were also significantly increased in the UGBE-treated group. The protein expression of toll-like receptor 4 was decreased by UGBE administration. Hematoxylin and eosin staining results also supported the results of this study showing normal appearance of liver histopathology in the UGBE-treated group. Conclusion: UGBE showed a great hepatoprotective effect on D-GalN/LPS-challenged rats via the toll-like receptor 4 signaling pathway.

Effects of Pretreated Korean Red Ginseng on Carbon Tetrachloride and Galactosamine-induced Hepatotoxicity in Rats (홍삼의 전처리에 의한 사염화탄소 및 갈락토사민 유발 간독성에 대한 보호효과)

  • Lee, Chung-Kyu;Kim, Na-Young;Han, Yong-Nam;Choi, Jong-Won
    • Journal of Ginseng Research
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    • v.27 no.1
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    • pp.1-10
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    • 2003
  • In present study, we examined whether or not the pretreatment of Korean Red Ginseng (KRG) could protect hepatotoxicity induced by carbon tetrachloride (CCl$_4$) and D-galatosamine (GalN). For this study, we not only tested activity of various plasma enzymes (AST, ALT, SDH, LDH), which are used as indicators of liver disease, but also checked the change of liver components such as lipid, glutathione and cytochromes content, and several liver enzyme activity. Pretreatment of KRG for two weeks significantly reduced the elevated plasma enzyme activities induced by CCl$_4$ and GalN. Pretreatment of KRG also restored the hepatic enzymes, malonedialdehyde formation, and depletion of reduced glutathione content induced by CCl$_4$ and GalN to near normal level. However, ${\gamma}$-glutamylcysteine synthetase activity was lot affected by KRG. These results suggest that KRG shows the hepatoprotective effect by reducing lipid peroxidation, by reducing the activity of free radical generating enzymes, and by preserving the hepatic glutathione.

Expression of the FLP recombinase of the 2 $\mu$m plasmid of yeast in the cultured cells of Bombyx mori using a transient expression vector (Yeast 2 $\mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현)

  • 강석우;윤은영
    • Journal of Sericultural and Entomological Science
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    • v.39 no.1
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    • pp.36-43
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    • 1997
  • In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

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Effect of Oral Administration of Fibrinolytic Enzyme from a Fermented Anchovy, Myulchi Jeot-Gal (멸치액젓 중 혈전용해효소의 경구 투여 효과)

  • 정영기;양웅석;김병기
    • Journal of Life Science
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    • v.8 no.6
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    • pp.737-740
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    • 1998
  • Effect of oral administration with fibrinolytic enzyme isolated from fermented anchovy(the traditional fermented food in Korea called Myulchi Jeot-gal) and its functionally active enzyme to rat, activation of plasma fibrinolysis was observed. The euglobulin fibrinolytic activities and the plasma levels of H-D-Val-Leu-Lys-pNA(S-2251) amidolysis reached a maximum at 3 hours after the administration to rat. And euglobulin Iysis time(ELT) value after oral admi-nistration showed its activity 2∼3 hours later. From the above result, it was confirmed the enzyme activity in blood by oral administration fibrinolytic enzyme through animal experiment.

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Changes of Ganglioside GM3 Expression in Porcine Oocyte Maturation and Early Embryonic Development In Vitro

  • Chae, Sung-Kyu;Park, Hyo-Jin;Kim, Jin-Woo;Ahn, Jae-Hyun;Park, Soo-Yong;Park, Jae-Young;Yang, Seul-Gi;Koo, Deog-Bon
    • Journal of Embryo Transfer
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    • v.30 no.4
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    • pp.319-325
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    • 2015
  • Gangliosides exist in glycosphingolipid-enriched domains on the cell membrane and regulate various functions such as adhesion, differentiation, and receptor signaling. Ganglioside GM3 by ST3GAL5 enzyme provides an essential function in the biosynthesis of more complex ganglio-series gangliosides. However, the role of gangliosides GM3 in porcine oocytes during in vitro maturation and early embryo development stage has not yet understood clear. Therefore, we examined ganglioside GM3 expression patterns under apoptosis stress during maturation and preimplantation development of porcine oocytes and embryos. First, porcine oocytes cultured in the NCSU-23 medium for 44 h after $H_2O_2$ treated groups (0.01, 0.1, 1 mM). After completion of meiotic maturation, the proportion MII (44 h) was significantly different among control and the H2O2 treated groups ($76.8{\pm}0.3$ vs $69.1{\pm}0.4$; 0.01 mM, $55.7{\pm}1.0$; 0.1 mM, $38.2{\pm}1.6%$; 1 mM, P<0.05). The expressions of ST3GAL5 in $H_2O_2$ treated groups were gradually decreased compared with control group. Next, changes of ST3GAL5 expression patterns were detected by using immunofluorescene (IF) staining during preimplantation development until blastocyst. As a result, we confirmed that the expressions of ST3GAL5 in cleaving embryos were gradually decreased (P<0.05) according to the early embryo development progress. Based on these results, we suggest that the ganglioside GM3 was used to the marker as pro-apoptotic factor in porcine oocyte of maturation and early embryo production in vitro, respectively. Furthermore, our findings will be helpful for better understanding the basic mechanism of gangliosides GM3 regulating in oocyte maturation and early embryonic development of porcine in vitro.