• Journal of Life Science
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• v.14 no.2
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• pp.315-319
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• 2004

For antibody development of human serine palmitoyltransferase (SPT, EC 2.3.1.50), SPTLC1 and SPTLC2 genes were subcloned in pRset vector and expressed in E. coli BL21 (DE3)pLys cells. Eucaryotic SPT is a membrane-bound heterodimer enzyme, while all other members are soluble homodimer enzymes. cDNA library were obtained from total RNA from human embryo kidney cell line, HEK293, using RT-PCR and PCR with specific primers was carried out for preparing SPTLC1 and SPTLC2 genes. pRset vector which can express hexahistidine-tag fusion protein was used and the DNA sequences of pRsetB/SPTLC1 and pRsetA/SPTLC2 were confirmed. Recombinant BL21 cells with SPTLC subunits were selected with LB plate containing ampicillin and chroramphenicol. SPTLC1 and SPTLC2 proteins were induced with 1 mM IPTG and seperated on 10% SDS-PAGE gel. Expressed proteins were confirmed by western blotting with His-tag antibody.

• Preventive Nutrition and Food Science
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• v.13 no.3
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• pp.196-203
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• 2008

Hwangtae, dried Alaska pollack, is a major storage product in the fish processing industry. Hwangtae is prepared by removing the internal organs and drying outdoors during the cold witner months by allowing it to thaw during the daytime and re-freeze at night under sub-zero ($-10^{\circ}C$) conditions and gradually dry from December until the next April for around 5 months from Myungtae. In this study, ground Hwangtae was hydrolyzed using two proteolytic enzymes (pepsin and alcalase) which produced five soluble active peptides from Hwangtae (yellowish dried Pollack, Theragra chalcogramma) protein. Two different peptides with strong antioxidative activity were isolated from the hydrolysate using consecutive chromatographic methods of Sephadex G-25 gel, ion-exchange chromatography on a Sepharose-Sephadex C-25 gel, and high-performance liquid chromatography. The isolated peptides, APO1 and APO2, were composed of 16 and 13 amino acid residues, respectively. Both peptides contained a Gly residue at the C-terminus and the repeating motif Gly-Pro-Hyp. The peptide with a molecular weight less than 1,000 Daltons (APACE) obtained from enzymatic hydrolysates of Hwangtae exhibited the highest ACE inhibitory activity. The APACE peptides was composed of 4 amino acid residues (Gly-Leu-Leu-Pro). These results suggest that Hwangtae hydrolysates could be a good source of peptides with ACE inhibitory activity. Biochemical analysis indicated that two 70 kDa peptides (APG1 and APG2) isolated from the hydrolysate had gelatinoytic activity, which was shown to be a calcium dependent protease type as showed by gelatin SDS PAGE.

• Korean Journal of Plant Resources
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• v.30 no.1
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• pp.88-95
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• 2017

Effects of different soil water conditions on fruit characteristics were investigated in 5-year-old 'Nero' black chokeberry trees (Aronia melanocarpa). Three kinds of drought stresses, including low water deficit, severe water deficit, and very severe water deficit, due to decline of soil water decreased the fruit quality of weight of 10 berries, soluble solid content, and anthocyanin, compared with the control (consistent water supply) during the harvest period. After longer drought stress, supply of soil water could induce berry cracking because cell size of epidermis of fruits contracted, whereas cell size of sub-epidermis and flesh expanded. Thus periodic water supply using water supply facility is needed for yield and quality of 'Nero' black chokeberry fruits.

• Archives of Pharmacal Research
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• v.28 no.2
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• pp.195-202
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• 2005

Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental factors are critical players in cellular damage and aging. In order to develop a new antiphotoaging agent, this work focused on the antioxidant effects of the extract of tinged autumnal leaves of Acer palmatum. One compound was isolated from an ethyl acetate soluble fraction of the A. palmatum extract using silica gel column chromatography. The chemical structure was identified as apigenin-8-C-beta-D-glucopyranoside, more commonly known as vitexin, by spectral analysis including LC-MS, FT-IR, UV, $^{1}H-$, and $^{13}C-NMR$. The biological activities of vitexin were investigated for the potential application of its anti-aging effects in the cosmetic field. Vitexin inhibited superoxide radicals by about $70\%$ at a concentration of $100\;{\mu}g/mL$ and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals by about $60\%$ at a concentration of $100\;{\mu}g/mL$. Intracellular ROS scavenging activity was indicated by increases in dichlorofluorescein (DCF) fluorescence upon exposure to UVB $20\;mJ/cm^2$ in cultured human dermal fibroblasts (HDFs) after the treatment of vitexin. The results show that oxidation of 5-(6-)chloromethyl-2',7'-dichlo-rodihydrofluorescein diacetate ($CM-H_{2}DCFDA$) is inhibited by vitexin effectively and that vitexin has a potent free radical scavenging activity in UVB-irradiated HDFs. In ROS imaging using a confocal microscope we visualized DCF fluorescence in HDFs directly. In conclusion, our findings suggest that vitexin can be effectively used for the prevention of UV-induced adverse skin reactions such as free radical production and skin cell damage.

• Archives of Pharmacal Research
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• v.26 no.5
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• pp.383-388
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• 2003

Bee venom is used as a traditional medicine for treatment of arthritis. The anti-inflammatory activity of the n-hexane, ethyl acetate, and aqueous partitions from bee venom (Apis mellifera) was studied using cyclooxygenase (COX) activity and pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production, in vitro. COX-2 is involved in the production of prostaglandins that mediate pain and support the inflammatory process. The aqueous partition of bee venom showed strong dose-dependent inhibitory effects on COX-2 activity ($IC_{50} = 13.1 \mu$ g/mL), but did not inhibit COX-1 activity. The aqueous partition was subfractionated into three parts by molecular weight differences, namely, B-F1 (above 20 KDa), B-F2 (between 10 KDa and 20 KDa) and BF-3 (below 10 KDa). B-F2 and B-F3 strongly inhibited COX-2 activity and COX-2 mRNA expression in a dose-dependent manner, without revealing cytotoxic effects. TNF-$\alpha and IL-1\beta$ are potent pro-inflammatory cytokines and are early indicators of the inflammatory process. We also investigated the effects of three subfractions on TNF-$\alpha and IL-1\beta$ production using ELISA method. All three subfractions, B-F1, B-F2 and B-F3, inhibited TNF-$\alpha and IL-1\beta$production. These results suggest the pharmacological activities of bee venom on anti-inflammatory process include the inhibition of COX-2 expression and the blocking of pro-inflammatory cytokines (TNF-$\alpha and IL-1\beta$) production.

• Journal of the East Asian Society of Dietary Life
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• v.26 no.6
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• pp.543-549
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• 2016

The objective of this study was to investigate quality properties of 'Cheonghyang' wine using different commercial yeast strains. Soluble solid content, pH level and total acidity of 'Cheonghyang' grape were $20^{\circ}Bx$, pH 3.5 and 0.66%, respectively. Total acidity ranged from 0.91~1.06% in the middle stage of fermentation and decreased to 0.77~0.82% when alcoholic fermentation finished. Alcohol content in wines ranged from 12.5% to 12.9% showing no significant difference in yeast strains. Wine fermented with Red Fruit had high volatile acid content (189.0 mg/L) whereas wine fermented with Fermivin indicated low volatile acid content (77.7 mg/L). Wines made with Montrachet, Fermivin and Aroma White had low brightness (L-value) compared to others. Results from sensory evaluation demonstrated that commercial wine yeasts, Montrachet and Fermivin, can be applied to improve sensory properties of 'Cheonghyang' wine such as aroma, acidity and transperency. On the other hand, preferences of wine fermented with EC-1118 strain containing lots of tannins and total polyphenols were significantly reduced.

• Molecules and Cells
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• v.40 no.5
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• pp.355-362
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• 2017

The ${\beta}2$ integrins are cell surface transmembrane proteins regulating leukocyte functions, such as adhesion and migration. Two members of ${\beta}2$ integrin, ${\alpha}M{\beta}2$ and ${\alpha}X{\beta}2$, share the leukocyte distribution profile and integrin ${\alpha}X{\beta}2$ is involved in antigen presentation in dendritic cells and transendothelial migration of monocytes and macrophages to atherosclerotic lesions. ${\underline{R}}eceptor$ for ${\underline{a}}dvanced$ ${\underline{g}}lycation$ ${\underline{e}}nd$ ${\underline{p}}roducts$ (RAGE), a member of cell adhesion molecules, plays an important role in chronic inflammation and atherosclerosis. Although RAGE and ${\alpha}X{\beta}2$ play an important role in inflammatory response and the pathogenesis of atherosclerosis, the nature of their interaction and structure involved in the binding remain poorly defined. In this study, using I-domain as a ligand binding motif of ${\alpha}X{\beta}2$, we characterize the binding nature and the interacting moieties of ${\alpha}X$ I-domain and RAGE. Their binding requires divalent cations ($Mg^{2+}$ and $Mn^{2+}$) and shows an affinity on the sub-micro molar level: the dissociation constant of ${\alpha}X$ I-domains binding to RAGE being $0.49{\mu}M$. Furthermore, the ${\alpha}X$ I-domains recognize the V-domain, but not the C1 and C2-domains of RAGE. The acidic amino acid substitutions on the ligand binding site of ${\alpha}X$ I-domain significantly reduce the I-domain binding activity to soluble RAGE and the alanine substitutions of basic amino acids on the flat surface of the V-domain prevent the V-domain binding to ${\alpha}X$ I-domain. In conclusion, the main mechanism of ${\alpha}X$ I-domain binding to RAGE is a charge interaction, in which the acidic moieties of ${\alpha}X$ I-domains, including E244, and D249, recognize the basic residues on the RAGE V-domain encompassing K39, K43, K44, R104, and K107.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.3
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• pp.81-90
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• 2007

Purpose: This study was conducted to investigate the muscle anti-fatigue effects of Allii tuberosi semen after long term administration. Materials and Methods: 4-6 weeks old ICR mice were used in this study and we administered the water soluble extracts of Allii tuberosi semen in the concentration of 1, 10 and 100mg/0.3ml to each 5 mice (ATS group) and same volume of normal saline was administered to 5 mice (control group) once a day for 90 or 120 days. After the administration we performed the swimming exercise test and the grip strength test. And we measured the concentration of glucose and the activity of lactate dehydrogenase in plasma regarded as biochemical indicator related with the muscular fatigue. Results: In swimming exercise test, all ATS groups showed increased time compared with relative control group after 90 days administration. But after 120 days adminstration 10 and 100mg/0.3ml ATS groups showed increased results. In grip strength test, all ATS group increased grip strength after 90 days administration and 10mg/0.3ml ATS group showed increased result after 120 days administration. All the results of plasma glucose didn't showed statistically significant difference after 90 and 120 days administration. In the plasma concentration of activity level of lactate dehydrogenase, 1mg/0.3ml and 10mg/0.3ml ATS groups showed decreased concentration compared with relative control group after 90 days administration. But after 120 days adminstration 1mg/0.3ml ATS groups showed decreased results. Conclusion: From this study we could find the anti-fatigue effect of Allii tuberosi semen in the muscle strength related test.

• Journal of Korean Society on Water Environment
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• v.37 no.2
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• pp.92-102
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• 2021

In this study, the distribution characteristics according to the type of BOD and the effect of nitrogenous oxygen demand (NOD) and algal oxygen demand on BOD in three streams (Bokhacheon, Byeongseongcheon, and Gulpocheon) were evaluated. Although the BOD and NOD concentrations demonstrated a difference in the three streams, the carbonaceous BOD(CBOD)/BOD ratio was 0.75 (p=0.053, one-way ANOVA), and there was no significant difference in the three streams (r2≥0.92, p<0.0001). The NOD concentration of the Bokhacheon with high NH3-N was 1.7±1.3 mg/L, which was the highest among the three streams and showed a significant correlation with BOD. Seasonal variations in NOD in the three streams did not show a significant correlation with changes in NH3-N concentration (r2<0.28, p≥0.1789), and there was no significant difference in NOD even though NH3-N concentration in Gulpocheon was about twice that of Byeongseongcheon (p=0.870, one way ANOVA). The particulate CBOD(PCBOD)/CBOD ratio of the three streams was 0.55~0.64, and about 60% of the biodegradable organic matter was present in the particulate form. When the Chl.a concentration in the stream was more than 7 ㎍/L, the PCBOD tended to increase with the Chl.a concentration (r2=0.61, p=0.003). In the three streams, particulate NOD accounted for 81% of NOD; however, despite the large variation in NH3-N concentration (0.075~3.182 mg/L), there was no significant difference in soluble NOD(SNOD) concentration that ranged from 0.1 to 0.3 mg/L. In this study, the low contribution rate of SNOD to NOD is considered as a result of the removal of nitrifying bacteria along with the particles during the filtration process.

• Journal of Applied Biological Chemistry
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• v.64 no.1
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• pp.97-102
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• 2021

In this study, enzymes were screened for hydrolysis of defatted perilla seed residue (DPSR) and optimal conditions for enzymatic treatment were determined to produce the hydrolysate of DPSR. Also its antioxidant activity and utilization as a culture medium were examined. The combined treatment of Alcalase and Ceremix is most effective for solubilization of protein and carbohydrate in DPSR. The optimal dosage, pH, and reaction time for enzymatic treatment were found to be 2.0% (w/w), 7.0, and 2 h, respectively. Treatment with optimal conditions of enzymes dramatically increased reducing sugar, soluble protein, and total phenolic content. The hydrolysate of DPSR possessed better scavenging activity against cation and free radicals than enzyme-untreated extract. When Leuconostoc mesenteroides 310-12 was cultured in the hydrolysate of DPSR, cell population rapidly increased compared to enzyme-untreated extract, and titratable acidity increased in proportion to the bacterial growth. In conclusion, these results imply that the hydrolysate of DPSR could be utilized as a bacteria culture medium as well as a physiologically active material with antioxidant activity.