• Applied Biological Chemistry
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• v.46 no.4
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• pp.299-304
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• 2003

A strain YB-9 was isolated from soil as a producer of the extracellular ${\beta}-D-galactosidase$, which catalyzes the hydrolysis of lactose. The strain YB-9 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supematant of the isolate with ammonium sulfate $(15{\sim}70%)$, the precipitated protein was used as a crude ${\beta}-galactosidase$ for analyzing its reaction properties with $para-nitrophenyl-{\beta}-D-galactoside$ $(pNP-{\beta}Gal)$ and lactose as substrates. The {\beta}-galactosidase showed its maximal activity at pH $6.0{\sim}6.5$ and $60^{\circ}C$. The hydrolyzing activity of ${\beta}-galactosidase$ for both $pNP-{\beta}Gal$ and lactose was decreased by galactose. Its hydrolyzing activity for lactose was slightly decreased by glucose, but the activity for $pNP-{\beta}Gal$ was increased to 1.3-folds by glucose. Especially, its hydrolyzing activity was not affected for lactose and was increased to 1.6-folds for $pNP-{\beta}Gal$ by xylose.

• Journal of Chest Surgery
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• v.45 no.6
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• pp.380-389
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• 2012

Background: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and ${\alpha}$-galactosidase (${\alpha}$-gal) to remove major xenoreactive antigens from xenogenic tissues. Materials and Methods: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) ${\alpha}$-gal or decellularization, or both were used to remove ${\alpha}$-gal from bovine pericardium. It was confirmed by ${\alpha}$-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or ${\alpha}$-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000${\times}$ transmission electron microscope (TEM). Results: Recombinant B. thetaiotaomicron ${\alpha}$-gal could effectively remove ${\alpha}$-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human ${\alpha}$-gal removed it recombinant human ${\alpha}$-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant ${\alpha}$-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant ${\alpha}$-gal made no difference in the arrangement of collagen fiber bundles with decellularization. Conclusion: Recombinant B. thetaiotaomicron ${\alpha}$-gal effectively removed ${\alpha}$-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant ${\alpha}$-gal, which may alleviate the harmful xenoreactive immunologic responses of ${\alpha}$-gal. Recombinant ${\alpha}$-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

• Journal of the Korean Geosynthetics Society
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• v.18 no.4
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• pp.107-114
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• 2019

In this paper, the P-wave multiple detection system for the fast and accurate earthquake early warning nearby the epicenter was developed. The developed systems were installed in five selected public buildings for the validation. During the monitoring, a magnitude 2.3 earthquake occurred in Pohang on 26 September 2019. P-wave initial detection algorithms were operated in three out of four systems installed in Pohang area and recorded as seismic events. At the nearest station, 5.5 km from the epicenter, P-wave signal was detected 1.2 seconds after the earthquake, and S-wave was reached 1.02 seconds after the P-wave reached, providing some alarm time. The maximum accelerations recorded in three different stations were 6.28 gal, 6.1 gal, and 5.3 gal, respectively. The alarm algorithm did not work, due to the high threshold of the maximum ground acceleration (25.1 gal) to operate it. If continuous monitoring and analysis are to be carried out in the future, the developed system could use a highly effective earthquake warning system suitable for the domestic situation.

• Current Research on Agriculture and Life Sciences
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• v.7
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• pp.153-163
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• 1989

The activity changes and biochemical properties of ${\beta}$-gal in tomato fruits during ripening were investigated. The total activity was increased during ripening and three isoenzymes (${\beta}$-gal I, II and III) were purified through DEAE Sephadex A-50 and Sephadex G-100 column chromatography. The activities of ${\beta}$gal isoenzymes (${\beta}$-gal I, II and III) during ripening were 69.8, 31.8 and 170.0 units in mature green phase, while those were 48.7, 88.4 and 136.8 units in Red phase, respectively. As the ripening proceeded the activities of ${\beta}$-gal I and III were some what decreased but the activity of ${\beta}$-gal II was incresed more than 2.8 fold. The optimum pH of ${\beta}$-gal I, II and III were 3.9, 4.2 and 3.9 and the optimum temperature of those were $60^{\circ}C$, $56^{\circ}C$ and $60^{\circ}C$, respectively. All isoenzymes were stable at pH 3.6~6.0 and lost their activity about 50% when it heated at $55^{\circ}C$ for 5 minute. $Mg^{{+}{+}}$-activated the three isoenzymes but $Ca^{{+}{+}}$ and SDS inhibited about 30~40%. $Hg^{{+}{+}}$ inhibited completely. The km value of ${\beta}$-gal I, II and III was 0.36mM, 0.63mM and 0.45mM, reaction rate was rapidly increased until the concentration of substrate was $6.0{\times}10^{-5}M$.

• Journal of Embryo Transfer
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• v.32 no.1
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• pp.17-24
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• 2017

Ganglioside GD1a is specifically formed by the addition of sialic acid to ganglioside GM1a by ST3 ${\beta}$-galactoside ${\alpha}$-2,3-sialyltransferase 2 (ST3GAL2). Above all, GD1a are known to be related with the functional regulation of several growth factor receptors, including activation and dimerization of epidermal growth factor receptor (EGFR) in tumor cells. The activity of EGF and EGFR is known to be a very important factor for meiotic and cytoplasmic maturation during in vitro maturation (IVM) of mammalian oocytes. However, the role of gangliosides GD1a for EGFR-related signaling pathways in porcine oocyte is not yet clearly understood. Here, we investigated that the effect of ST3GAL2 as synthesizing enzyme GD1a for EGFR activation and phosphorylation during meiotic maturation. To investigate the expression of ST3GAL2 according to the EGF treatment (0, 10 and 50 ng/ml), we observed the patterns of ST3GAL2 genes expression by immunofluorescence staining in denuded oocyte (DO) and cumulus cell-oocyte-complex (COC) during IVM process (22 and 44 h), respectively. Expression levels of ST3GAL2 significantly decreased (p<0.01) in an EGF concentration (10 and 50 ng/ml) dependent manner. And fluorescence expression of ST3GAL2 increased (p<0.01) in the matured COCs for 44 h. Under high EGF concentration (50 ng/ml), ST3GAL2 protein levels was decreased (p<0.01), and their shown opposite expression pattern of phosphorylation-EGFR in COCs of 44 h. Phosphorylation of EGFR significantly increased (p<0.01) in matured COCs treated with GD1a for 44 h. In addition, ST3GAL2 protein levels significantly decreased (p<0.01) in GD1a ($10{\mu}M$) treated COCs without reference to EGF pre-treatment. These results suggest that treatment of exogenous ganglioside GD1a may play an important role such as EGF in EGFR-related activation and phosphorylation in porcine oocyte maturation of in vitro.

• Reproductive and Developmental Biology
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• v.39 no.4
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• pp.127-132
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• 2015

To overcome the hyperacute immune rejection during pig-to-non-human primates xenotranasplantation, we have produced and bred ${\alpha}$-1,3-galactosyltransferase knock-out ($GalT^{-/-}$) pigs. In this study, the somatic cells and tissues from the $GalT^{-/-}$ pigs were characterized by an analysis of the expression of Gal${\alpha}$-1,3-Gal (${\alpha}-Gal$) epitope. Briefly, ear fibroblast cell lines of 19 homozygous $GalT^{-/-}$ pigs were established and cryopreserved. The expression of ${\alpha}-Gal$ epitope in the cells was measured by fluorescence activated cell sorter (FACS) analysis using BS-I-B4 lectin. Also, the homozygous ($GalT^{-/-}$) cells and tissues samples were immunostained with BS-I-B4 lectin for analysis of ${\alpha}-Gal$ epitope expression. The results showed that the expression of ${\alpha}-Gal$ epitope in $GalT^{-/-}$ cells (0.2 %) were significantly (p<0.05) down-regulated to the range of cynomolgus monkey fibroblast (0.2 %) cells compared to heterozygous ($GalT^{-/+}$) (9.3 %) and wild type ($GalT^{+/+}$) (93.7 %) fibroblast cells. In the immunostaining results, while the expression of ${\alpha}-Gal$ epitope was detected a partly in $GalT^{-/+}$ cells and mostly in $GalT^{+/+}$ cells, it was almost not detected in the $GalT^{-/-}$ cells. Also, immunostaining results from various tissues of the $GalT^{-/-}$ pig showed that the expression of ${\alpha}-Gal$ epitope was not detectable, whereas various tissues from $GalT^{+/+}$ pig showed a strong expression of ${\alpha}-Gal$ epitope. Our results demonstrated that ${\alpha}-Gal$ epitope expressions from $GalT^{-/-}$ pigs were successfully knocked out to prevent hyperacute immune rejection for further study of xenotransplantation.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.662-666
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• 2019

In this study, the NABH558 gene expression system was constructed to efficiently produce neoagarobiose hydrolase (NABH) in Saccharomyces cerevisiae strain. The ADH1 and GAL10 promoters of the pAMFα-NABH and pGMFα-NABH plasmids were examined to determine the suitable promoter for the NABH558 gene expression, respectively. The effect of promoter and carbon sources on NABH558 gene expression was investigated by transforming each plasmid into the S. cerevisiae 2805 strain. The NABH activity in the 2805/pAMFα-NABH strain was 0.069 unit/ml/DCW in YPD medium, whereas that in the 2805/pGMFα-NABH strain was similar (0.02-0.027 unit/ml/DCW) irrespective of the medium composition. The higher NABH activity in the YPD medium was due to the increased NABH558 gene transcription. NABH produced in the recombinant strains could degrade agarose to galactose and AHG. This indicated that ADH1 promoter was a more optimal promoter for the expression of NABH558 gene than the GAL10 promoter. The NABH activity induced by the ADH1 promoter was about 3-fold higher than that induced by the GAL10 promoter.

• Environmental Mutagens and Carcinogens
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• v.22 no.3
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• pp.175-182
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• 2002

Endocrine disruptors (EDs) are the chemicals that affect endocrine systems through activation or inhibition of steroid hormone response. It is necessary to have a good system to evaluate rapidly and accurately endocrine-disrupting activities of suspected chemicals and their degradation products. The key targets of EDs are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. We constructed a co-expression system of Gal4p DNA binding domain (DBD)- ligand binding domain of human estrogen receptor $\alpha$ or $\beta$, and Gal4p transactivation domain (TAD)-co-activator AIB-1, SRC-1 or TIF-2 in Saccharomyces cerevisiae with a chromosome-integrated lacZ reporter gene under the control of CYC1 promoter and Gal4p binding site (GAL4 upstream activating sequence, GAL4$_{UAS}$). Expression of this reporter gene was dependent on the presence of estrogen or EDs in the culture medium. We found that the two-hybrid system with combination of the hER$\beta$ LBD and co-activator SRC-1 was most effective in the xenoestrogen-dependent induction of reporter activity. The extent of transcriptional activation by those chemicals correlated with their estrogenic activities measured by other assay systems, indicating that this assay system is efficient and reliable for measuring estrogenic activity. The data in this research demonstrated that the yeast detection system using steroid hormone receptor and co-activator is a useful tool for identifying chemicals that interact with steroid receptors.s.

• KSBB Journal
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• v.14 no.4
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• pp.476-481
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• 1999

S. cerevisiae mutant(reg1-501, gal1), which cannot use galactose and has alleviated glucose repression level, is used as host for optimizing induction of GAL promoter. The optimum concentration of galactose as inducer for recombinant protein production and the galactose consumption rate have been tested with S. cerevisiae mutant and compared with conventional S. cerevisiae. The extent of glucose repression were investigated for both strain and the degradation pattern of produced foreign protein have been compared in both cases. The effect of pH on foreign protein degradation pattern were studied for both strains. The secetion efficiency of both strains were carried out. Through these experiments, optimum condition of recombinant protein production by GAL promoter using S. cerevisiae mutant (reg1-501, gal1) were found.

• Journal of Ginseng Research
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• v.39 no.4
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• pp.376-383
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• 2015

Background: Panax ginseng has a wide range of biological activities including anti-inflammatory, antioxidant, and immunomodulatory functions. Wild ginseng cambial meristematic cells (CMCs) were obtained from P. ginseng cambium. This study examined the protective mechanism of wild ginseng CMCs against $\small{D}$-galactosamine (GalN)-induced liver injury. GalN, a well-known hepatotoxicant, causes severe hepatocellular inflammatory damage and clinical features similar to those of human viral hepatitis in experimental animals. Methods: Hepatotoxicity was induced in rats using GalN (700 mg/kg, i.p.). Wild ginseng CMCs was administered orally once a day for 2 wks, and then 2 h prior to and 6 h after GalN injection. Results: Wild ginseng CMCs attenuated the increase in serum aminotransferase activity that occurs 24 h after GalN injection. Wild ginseng CMCs also attenuated the GalN-induced increase in serum tumor necrosis factor-${\alpha}$, interleukin-6 level, and hepatic cyclooxygenase-2 protein and mRNA expression. Wild ginseng CMCs augmented the increase in serum interleukin -10 and hepatic heme oxygenase-1 protein and mRNA expression that was induced by GalN, inhibited the increase in the nuclear level of nuclear factor-kappa B, and enhanced the increase in NF-E2-related factor 2. Conclusion: Our findings suggest that wild ginseng CMCs protects liver against GalN-induced inflammation by suppressing proinflammatory mediators and enhancing production of anti-inflammatory mediators.