• Korean Journal of Food Science and Technology
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• v.45 no.5
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• pp.619-627
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• 2013

We investigated the characteristics and biological activity of makgeolli supplemented with different levels (0%, 1%, 3%, 5%, and 7%) of Akebia quinata fruit during fermentation. Our results showed that supplementation with Akebia quinata fruit led to an increase in the acidity level, amino acid concentration, alcohol content, and total sugar level. Makgeolli supplemented with 7% Akebia quinata fruit showed the highest total sensory score. Supplementation with Akebia quinata fruit resulted in a significant increase in the antioxidant activity and nitric oxide inhibitory activity. Further, makgeolli supplemented with Akebia quinata fruit showed anticancer activity against DU145, HeLa, MCF-7, and U87cells, and significantly enhanced antibacterial activity against Shigella flexneri. Our results indicate that Akebia quinata fruit represents an effective natural additive for enhancing the biological activities of makgeolli.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.1
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• pp.1-7
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• 2008

The antioxidative, antimutagenic and cytotoxic activities of ethanol extract from Cornus officianalis have been studied. The antioxidant activity of the ethanol extract was measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. The inhibition effects on the mutagenicity in Salmonella Typhimurium TA100 were evaluated by Ames test and cancer cell inhibitory effects in Hep3B cell and HeLa cell were tested by MTT assay. Cornus officianalis had an important free radical-scavenging activity towards the DPPH radical. At a concentration of 500 ppm, the DPPH radical-scavenging activity of Cornus officianalis was similar to that of L-ascorbic acid. None of the extracts produced a mutagenic effect on S. Typhimurium TA100. The ethanol extract from Cornus officianalis showed about 77% of inhibition at 500 ppm on the mutagenicity induced by 4-nitroquinoline-1-oxide. The extract from Cornus officianalis showed strong cytotoxicity against Hep3B and HeLa cells, with inhibition of 83 and 78% at a dose of $700{\mu}g$/plate, respectively. Moreover, the ethanol extracts had 34.33 mg H.E/g of polyphenols and 5.67 mg Q.E/g of flavonoids, respectively. Therefore, the present study showed antioxidative, antimutagenic and anticancer potential of the ethanol extract from Cornus officianalis.

• Molecules and Cells
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• v.24 no.3
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• pp.378-387
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• 2007

The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.27-27
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• 2003

A diversified and concentrative approach of methylation player can be one of the most powerful studies in the understanding of global epigenetic modifications. Previous studies have suggested that DNA methylation contributes to transcriptional silencing through the several DNA methylation-mediated repression systems by hypermethylation, including methyltransferases (DNMTs), DNA methyltransferase association protein 1 (DMAPl), methyl-CpG binding domain (MBD), and histone deacetylases (HDACs). Assembly of these regulatory protein complexes act sequentially, reciprocally, and interdependently on the newly composed DNA strand through S phase. Therefore, these protein complexes have a role in coupling DNA replication to the designed turn-off system in genome. In this study, we attempted to address the role of DNA methylation by the functional analysis of the methyltransferase molecule, we described the involvement of DMAP1 and DNMTs in cell divistion and the effect of their loss. We also described distinct patterns that DMAP1 and DNMTs are spatially reorganized and displaced from condensing chromosomes as cells progress through mitosis in HeLa cell, COS7, and HIH3T3 cell cycle progressions. DNMT1, DNMT3b, and DMAP1 do not stably contact the genetic material during chromosome compaction and repressive expression. These finding show that the loss of activities of DNMTs and DMAP1 occure stage specifically during the cell cycle, may contribute to the integral balance of global DNA methylation. This is consistent with previous studies resulted in decreased histone acetyltransferases and HDACs, and differs from studies resulted in increased histone methyltransferases. Our results suggest that DNA methylation by DNMTs and DMAP1 during mitosis acts to antagonize hypermethylation by which this mark is epigenetical mitotic-specific methylation.

• Journal of Microbiology and Biotechnology
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• v.31 no.6
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• pp.855-866
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• 2021

The effects of various carbon sources on mycelial growth and polysaccharide synthesis of the medicinal fungus Inonotus obliquus in liquid fermentation were investigated. After 12-d fermentation, mycelial biomass, polysaccharide yield, and polysaccharide content were significantly higher in Glc+Lac group (glucose and lactose used as combined carbon source) than in other groups. Crude polysaccharides (CIOPs) and the derivative neutral polysaccharides (NIOPs) were obtained from mycelia fermented using Glc, fructose (Fru), Lac, or Glc+Lac as carbon source. Molecular weights of four NIOPs (termed as NIOPG, NIOPF, NIOPL, and NIOPGL) were respectively 780.90, 1105.00, 25.32, and 10.28 kDa. Monosaccharide composition analyses revealed that NIOPs were composed of Glc, Man, and Gal at different molar ratios. The NIOPs were classified as α-type heteropolysaccharides with 1→2, 1→3, 1→4, 1→6 linkages in differing proportions. In in vitro cell proliferation assays, viability of RAW264.7 macrophages was more strongly enhanced by NIOPL or NIOPGL than by NIOPG or NIOPF, and proliferation of HeLa or S180 tumor cells was more strongly inhibited by NIOPG or NIOPGL than by NIOPF or NIOPL, indicating that immune-enhancing and anti-tumor activities of NIOPs were substantially affected by carbon source. qRT-PCR analysis revealed that expression levels of phosphoglucose isomerase (PGI) and UDP-Glc 4-epimerase (UGE), two key genes involved in polysaccharide synthesis, varied depending on carbon source. Our findings, taken together, clearly demonstrate that carbon source plays an essential role in determining structure and activities of I. obliquus polysaccharides by regulating expression of key genes in polysaccharide biosynthetic pathway.

• Biomolecules & Therapeutics
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• v.17 no.4
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• pp.430-437
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• 2009

Metabolites of Soyasaponin I, a major constituent of soybean, by human intestinal microflora were investigated by LC-MS/MS analysis. We found four peaks, one parental constituent and three metabolites: m/z 941 [M-H]$^-$, m/z 795 [M-rha-H]$^-$, m/z 441 [aglycone-$H_2O$+H]$^+$, and m/z 633 [M-rha-gal-H]$^-$, which was an unknown metabolite, soyasapogenol B 3-$\beta$-D-glucuronide. When soyasaponin I was incubated with the human fecal microbial fraction from ten individuals for 48 h, soyasaponin I was metabolized to soyasapogenol B via soyasaponin III and soyasapogenol B 3-$\beta$-D-glucuronide or via soyasaponin III alone. Both soyasaponin I and its metabolite soyasapgenol B exhibited estrogenic activity. Soyasaponin I increased the proliferation, mRNA expression of c-fos and pS2, in MCF7 cells more potently than soyasapogenol B. However, soyasapogenol B showed potent cytotoxicity against A549, MCF7, HeLa and HepG2 cells, while soyasaponin I did not. The cytotoxicity of soyasapogenol B may prevent its estrogenic effect from increasing dose-dependently. These findings suggest that orally administered soyasaponin I may be metabolized to soyasapogenol B by intestinal microflora and that soyasapogenol B may express a cytotoxic effect rather than an estrogenic effect.

• Korean Journal of Microbiology
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• v.38 no.2
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• pp.86-95
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• 2002

As a result of genome projects, the research to elucidate the function of a protein of interest has recently been well-recognized. In order to facilitate functional genomics, a useful mammalian gene expression vector is required. Using an infectious CDNA clone of BVDV pNADLclns-, we have developed a mammalian gene expression vector. In this study, a replication-competent full-length infectious CDNA clone containing puremycin acetyltransferase (pac) gene (pNADLclns-/pac) was successfully generated. The viral RNA replication and viral protein NS3 synthesis were examined by detecting metabollically $^{32}P$-labelled genomic viral RNA and immunoblotting with a mouse anti-NS3 antibody. To generate viral replicon as an expression vector, we examine if the viral structural genes (C, E0, El, E2) are required for viral replication by deletion analysis. As a result, all of the structural proteins are dispensable for viral replication per se, but essential for infectious viral particle formation. Based on our deletion analysis, we have generated a replication-competent BVDV viral replicon (pNADLclns-/pac/${\Delta}S$), whose structural genes are all deleted. In addition to NADLclns- /pac/${\Delta}S$, NADLclns-/ luc/${\Delta}S$ viral replicon containing luciferase gene as a reporter was constructed and fecund to be replication-compotent in HeLa and BHK cells as well as MDBK cells. Therefore, BVDV viral replicon developed in our study will be a useful tool to express a protein of interest in various mammalian cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.29 no.5
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• pp.922-934
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• 2000

To investigate the composition of carotenoids present in marine organisms and the biological activity of the carotenoids, carotenoids of the muscles and tunic of tunicates and shellfishes were isolated and identified. Anitmutagenic activities of the carotenoids for S. typhimurium TA 98 and cytotoxic activity for cancer cell lines were determined. Total carotenoid contents in the muscle of tunicata ranged from 18.65 mg% to 2.39 mg%. The highest amount of the total carotenoid was found in the muscle of Halocynthia aurantium, followed by Styela clava (HERDMAN), H. roretzi, H. hilgendorfi f. igaboya, H. hilgendorfi f. retteri, S. plicata (LESUEUR) in order. Interestingly, total carotenoid content in the muscle of S. clava (HERDAMAN) was higher than that of H. roretzi. Total carotenoid content of all tunicata, other than H. aurantium and H. roretzi, were higher in muscle than tunic. The major carotenoids in H. roretzi, H. aurantium, S. plicata (LESUEUR), and S. clava (HERDAMAN) were cynthiaxanthin (25.1∼42.2%), halocynthiaxanthin (9.7∼26.3%), diatoxanthin (8.0∼18.7%) and β-carotene (7.7%∼21.7%). Similarly, cantaxanthin (19.6%), cynthiaxanthin (15.4%), halocynthiaxanthin (14.8%), and (3R, 3'R), (3S, 3'S)-astaxanthin (22.6%) in H. hilgendorfi f. retteri and fucoxanthin (26.6%), cynthiaxanthin (21.8%), halocynthiaxanthin (15.2%), and β-carotene (9.3%) in H. hilgendorfi f. igaboya were major carotenoids in both tunicate. However, the composition of carotenoids in muscle and tunic of tunicata was similar each other. Among the shellfishes examined, total carotenoid content of the muscle of Peronidia venulosa (Schrenck) and Corbicula fluminea, and of the gonad of Atrina pinnata and Chlamys farreri, was ranged from 2.51 to 6.83 mg% which were relatively higher than that of other shellfishes. The composition of the carotenoids of shellfishes, which might depend upon their living environments, was varied. But cynthiaxanthin (15.9∼39.0%) and zeaxanthin (9.6∼21.9%) in gonad of C. farreri, and muscles of Buccinum Volutharpa perryi (JAY) and Crassostrea gigas, cynthiaxanthin (21.5∼48.6%) and mytiloxanthin (14.6%) in muscle of C.fluminea and gonad of A. pinnata, and canthaxanthin (60.6%) and isozeaxanthin (20.5%) in muscles of P. venulosa (Schrenck), and β-carotene (23.7%∼37.8%) and zeaxanthin (18.2∼20.4) in muscles of Semisulcospira libertina and Meretrix lusoria were major carotenoids. Interestingly, diester type-carotenoids were present along with free type-carotenoids in muscles of C. gigas. antimutagenic effect of the carotenoids isolated from tunicata and shellfishes against 2-amino-3-methylimidazol [4,5-f]quinoline (IQ) for S. typhimurium TA 98 was proportional to the amount (20, 50 and 100㎍/plate) treated. Mutagenicity of IQ was significantly reduced by astaxanthin, isozeaxanthin, mytiloxanthin and halocynthiaxanthin, whereas the mutagenicity of aflatoxin B₁(AFB₁) was significantly reduced by β-carotene, isozeaxanthin, and mytiloxnthin. Growth inhibition effect of carotenoids isolated from tunicata and shellfishes for cancer cell was proportional to the amount (5, 10, and 20㎍/plate) treated. The growth of HeLa cell by β-carotene, cynthiaxanthin, astaxanthin and halocynthiaxanthin, NCI-H87 cell by β-carotene, astaxanthin, cynthiaxanthin, and halocynthiaxanthin, HT-29 cell by β-carotene, cynthiaxanthin, mytiloxanthin and halocynthiaxanthin, and MG-63 cells by β-carotene, cynthiaxanthin, astaxanthin, canthaxanthin and halocynthiaxanthin were statistically reduced.

• Food Science and Preservation
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• v.14 no.2
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• pp.220-225
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• 2007

This study was performed to measure the antioxidative, antimutagenic, and cytotoxic properties of Prunus armeniaca using the DPPH (1, 1-diphenyl-2-picrylhydrazyl) free radical donating method, the Ames test, and cytotoxicity measurements, respectively. Electron-donating abilities were 48.3, 43.9, 14.8 and 12.9 per g dry matter of P. armeniaca seed (PAS), P. armeniaca flesh(PAF), butylated hydroxytoluene, and ${\alpha}-tocopherol$, respectively. The direct antimutagenic effects of an ethanol extract of P. armeniaca were examined in Ames tests using Salmonella typhimurium TA98 and TA100 as reporter organisms. In the Ames test, the ethanol extract of P. armenicaca alone did not exhibit any mutagenicity but the extract did show substantial inhibitory effects against mutations induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide(4NQO). The ethanol extract of PAS(200g dry matter/plate) inhibited strain TA98 mutagenesis induced by 4NQO by ca. 37.9%, and mutation inhibition values of 42.1% and 69.4%, respectively, were observed when 4NQO and MNNG acted on the TA100 strain. The cytotoxic effects of ethanol extracts of P. armeniaca against cell lines of human lung carcinoma(A549), human breast adenocarcinoma(MCF-7), human hepatocellular carcinoma(Hep3B), human cervical adenocarcinoma(HeLa), and human gastric carcinoma(AGS) rose with increases in extract concentration. An ethanol extract(4mg/mL dry matter) of PAF showed strong cytotoxicities of 88.2%, 58%, 72.8%, 89.4%, and 91.9% against A549, AGS, MCF-7, HeLa, and Hep3B cells, respectively. In contrast, the same extract showed only 13 37% cytotoxicity for a nomal human kiney cell line(293). It is suggested that P. armeniaca possesses useful antioxidative, antimutagenic, and anticancer properties.

• Korean Journal of Microbiology
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• v.37 no.3
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• pp.221-227
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• 2001

Heat shock proteins serve as chaperone by preventing the aggregation of denatured proteins and promote survival of pathogens in harsh environments. In bacteria, ethanol shock induced the major chaperone GroEL and DnaK, but in Streptococcus pneumoniae, it induced neither GroEL nor DnaK but alcohol dehydrogenase (ADH). In this study, ADH gene encoding a 104-kDa (p104) protein was identified and characterized. The deduced amino acid sequence of pneumococcal ADH shows homology with other members of the ADH family, and particularly with Entamoeba histolytica ADH2 and E. coli ADH. S. pneumoniae adh is composed of 883 amino acids and its estimated isoelectric point is 6.09. Although ADH is conserved between S. pneumoniae and E. coli, immunoblot analysis employing antisera raised against pneumococcus ADH demonstrated no cross-reactivity with ADH analog in Eschericha coli, Staphylococcus aureus and human HeLa cells. Also secretion of ADH was demonstrated by subcellular fractionation and immunoblot analysis of proteins. These results suggest that S. pneumoniae ADH could be a highly feasible candidate for both diagnostic marker and vaccine.