• KSBB Journal
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• v.18 no.4
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• pp.255-260
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• 2003

In vivo bioassays for biological medicines have been considered final resort to unequivocally assess the biological activities for them because there are some cases in which the biological activities obtained from in vivo bioassay and in vitro bioassay quite differ each other. The in vivo biological activity of EPO depends on its sialic acid contents which confer microheterogeneity-isoforms to this protein. We have devise a method which consists of a in vitro bioassay using BaF3 cell line and a capillary zone electrophoresis (CZE) for the measurement of the EPO isoform distribution. The biological activity of EPO obtained using in vitro bioassay with BaF3 cell line showed good correlation (C.V.(％) 7.34, 5.85, 8,16, 8.08, 8.8) to EPO content measured either spectrophotometric assay (A280 0.1 ％ ＝0.743) or radio immunoassay. The assay validation results of in vitro bioassay with 3 lot of in house EPO showed good results to EPO content measured either in vivo assay or radio immunoassay. and also showed good results the robustness of our method in terms of precision, accuracy, repeatability. The isoform distribution for EPO-BRP (1 : 1 mixture of epoetin-${\alpha}$ and epoetin-${\beta}$, European Pharmacopoeia) by CZE method resulted in isoform 2 through isoform 8. The major peaks in electrophoregram were composed of isoform 3 through 7. Our recombinant EPO (epoetin-${\alpha}$) having equivalent in vivo biological activity showed the isoform distribution of isoform 3 through 9. The major peaks consisted of isoform 4 through 8. The peak area of isoform 4 was always smaller than that of isoform 5. The preparations of recombinant epoetin-${\alpha}$ with lower in vivo biological activity than EPO-BRP showed the isoform 2 through 8 in their electrophoregrams whose major peaks consisted of the isoform 3 through 7. The peak area of isoform 4 was larger than that of isoform 5.

• Restorative Dentistry and Endodontics
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• v.25 no.2
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• pp.170-179
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• 2000

The rh-BMP-4 is a subgroup of TGF-${\beta}$ superfamily. The application of rh-BMP in alveolar bony defect was reported to new alveolar bone and new cementum formation. For minimized complications following tooth replantation, a operator must replant a tooth fast at the pertinent position. This study was to evaluate the effect of rh-BMP-4 on periodontal regeneration and root resorption following tooth replantation in rats. The 50 Sprague-Dawley rats weighting about 130gm were used in this study. The animals were divided into three groups. Group 1 ; immediate replantation after extraction : Group 2 ; replantation stored teeth extraction of first molar, the removal of periodontal ligament with collagenase, and etching with citric acid : Group 3 ; replantation stored teeth with treated rh-BMP-4 in mesial root. Experimental animals were sacrificed 3, 7, 14 days after replantation by heart infusion. The maxillae were removed, fixed, demineralized, dehydrated, infiltrated and embedded with JB-4 mixture. For light microscopic observation, 5 micron sections were cut and stained with toluidine blue. The results of this study were as follows : 1. After experimental 3 days, all groups were observed dead space between periodontum and root. 2. After experimental 7 days, group 1 and group 3 were observed filling periodontal fibers between alveolar bone and root but group 2 were not. 3. After experimental 7 days, group 3 were observed appearance of attached cementoblast like cell on root surface. Group 1 were observed regular arrangement of fibroblasts and collagen fibers at ${\times}400$ observation. 4. After experimental 14 days, all group were observed filling periodontal fibers between alveolar bone and root. Group 1 were observed normal arrangement of periodontal fibers. Group 3 were observed less abnormal arrangement of periodontal fibers. Group 2 were not observed functional normal arrangement of periodontal fibers. 5. After experimental 14 days, group 2 and 3 were observed several root resorption and irregular root surface but group 1 were not. These results suggest that the rh-BMP-4 can stimulate cementogenesis and enhance to attach collagen fibers.

• Herbal Formula Science
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• v.26 no.4
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• pp.295-306
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• 2018

Schisandra chinensis (SC) and Lycium chinense (LC) were widely distributed in Asia and the fruit has been used traditionally for medicinal herbs. The processing method was solid-state fermentation using Aspergillus oryzae for 48 h after stir-frying treatment at $220^{\circ}C$ for 12 min. In this study, in vitro the anti-inflammatory effect and in vivo hangover reduction were compared to unprocessed SC and LC water extract. Anti-inflammatory effects have been evaluated in pro-inflammatory mediators which were secreted by lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Nitric oxide (NO) was determined using Griess reaction. Proinflammatory cytokines such as tumor necrosis factor $(TNF)-{\alpha}$ and interleukin $(IL)-1{\beta}$ were measured by enzyme-linked immunosorbent assays (ELISA). Alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ALDH) activities were compared to processed SC or LC and mixtures thereof (1:1). In vivo study was compared to hangover relief in alcohol-fed mice. After administering a mixture of SC and LC (300 mg/kg) water extract (1:1), mice were fed 3 g/kg of ethanol. Serum was collected at 1, 3, and 5 h intervals to analyze ethanol and acetaldehyde levels using a colorimetric assay kit. The processed SC and LC water extracts compared to raw materials significantly inhibited LPS-induced NO and inflammatory cytokine production in RAW 264.7 cells. The results of the hangover mouse model are also consistent with anti-inflammatory effects. These results suggest that processed SC and LC extracts may be functional materials for the treatment of inflammation and hangover.

• Journal of Applied Biological Chemistry
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• v.62 no.3
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• pp.229-237
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• 2019

More effective treatments are needed for non-alcoholic fatty liver disease (NAFLD). We hypothesized that water extracts of blackberry fruits (BF) and leaves (BL) and their combinations (BFL) reduce fat deposition in HepG2 cells and modulate shor-tchain fatty acids (SCFA) and fecal bacteria in vitro. HepG2 cells were treated with BF, BL, BFL1:2, and BFL1:3 for 1 h, and 0.5 mM palmitate was added to the cells. Moreover, low ($30{\mu}g/mL$) and high doses ($90{\mu}g/mL$) of BL and BF were applied to fecal bacteria in vitro, and SCFA was measured by GC. BL, BF, BFL1:2, and BFL1:3 reduced triglyceride deposition in the cells in a dose-dependent manner, and BFL1:2 and BFL1:3 had a stronger effect than BF. The content of malondialdehyde, an index of oxidative stress, was also reduced in BL, BF, and BFL1:2 with increasing superoxide dismutase and glutathione peroxidase activities. The mRNA expression of acetyl CoA carboxylase, fatty acid synthase, and sterol regulatory element-binding protein-1c was reduced in BL, BF, BFL1:2, and BFL1:3 compared to the control, and BFL1:2 had the strongest effect. By contrast, the carnitine palmitolytransferase-1expression, a regulator of fatty acid oxidation, increased mostly in BFL1:2 and BFL1:3. Tumor necrosis factor-${\alpha}$ and interleukin-$1{\beta}$ expression was reduced in BL compared to that in BF and BFL1:2 in HepG2 cells. Interestingly, BL increased propionate production, and BF increased butyrate and propionate production and increased total SCFA content in fecal incubation. BF increased the contents of Bifidobacteriales and Lactobacillales and decreased those of Clostridiales, whereas BL elevated the contents of Bacteroidales and decreased those of Enterobacteriales. In conclusion, BFL1:2 and BFL1:3 may be potential therapeutic candidates for NAFLD.

• Journal of the Korean Applied Science and Technology
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• v.38 no.1
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• pp.99-106
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• 2021

To circumvent the skin problem from phenylethanol (PhE), we have studied on the enzymatic synthesis of phenylethanol galactoside (PhE-gal) as an alternative to PhE. Base on the previous study, we optimized the reaction conditions for PhE-gal synthesis from PhE using E. coli β-galactosidase (β-gal). The optimal amount of β-gal, PhE concentration, pH, and temperature for PhE-gal synthesis were 0.45 U/ml, 1%, 8.0, 40℃, respectively. Under these conditions, about 81.9 mM PhE was converted into about 47.4 mM PhE-gal, in which the conversion yield was about 57.9%. Meanwhile, when the reaction mixture containing PhE and PhE-gal was mixed and fractionated with water-immiscible solvent (EA or MC), it was observed that PhE-gal was distributed in water phase, and PhE was distributed in solvent phase. Additionally, PhE-gal was clearly distributed into water phase when MC was used, but PE-gal was not when EA was used. In the future, we are planning to carried out the continuing study on developing an alternative cosmetic preservative using PhE-gal.

• Proceedings of the Materials Research Society of Korea Conference
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• 2003.03a
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• pp.81-81
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• 2003

The stochiometric mixture of evaporating materials for the CuGaSe$_2$ single crystal thin films were prepared from horizontal furnace. Using extrapolation method of X-ray diffraction patterns for the polycrystal CuGaSe$_2$, it was found tetragonal structure whose lattice constant no and co were 5.615$\AA$ and 11.025$\AA$, respectively. To obtains the single crystal thin films, CuGaSe$_2$ mixed crystal was deposited on throughly etched GaAs(100) by the Hot Wall Epitaxy(HWE) system. The source and substrate temperature were 61$0^{\circ}C$ and 45$0^{\circ}C$ respectively, and the growth rate of the single crystal thin films was about 0.5${\mu}{\textrm}{m}$/h. The crystalline structure of single crystal thin films was investigated by the double crystal X-ray diffraction(DCXD). Hall effect on this sample was measured by the method of van der pauw and studied on carrier density and mobility depending on temperature. From Hall data, the mobility was likely to be decreased by pizoelectric scattering in the temperature range 30K to 150K and by polar optical scattering in the temperature range 150K to 293K. The optical energy gaps were found to be 1.68eV for CuGaSe$_2$ single crystal thin films at room temperature. The temperature dependence of the photocurrent peak energy is well explained by the Varshni equation then the constants in the Varshni equation are given by a=9.615$\times$ 10$^{-4}$ eV/K, and $\beta$=335K. From the photocurrent spectra by illumination of polarized light of the CuGaSe$_2$ single crystal thin films. We have found that values of spin orbit coupling ΔSo and crystal field splitting ΔCr was 0.0900eV and 0.2498eV, respectively. From the PL spectra at 20K, the peaks corresponding to free bound excitons and D-A pair and a broad emission band due to SA is identified. The binding energy of the free excitons are determined to be 0.0626eV and the dissipation energy of the acceptor-bound exciton and donor-bound exciton to be 0.0352eV, 0.0932eV, respectively.

• Animal Bioscience
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• v.36 no.6
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• pp.869-878
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• 2023

Objective: Hyperketonemia remains a major metabolic issue of serious milk production and a major health concern in early lactation cows. Oral supplementation of glucose precursors (GP) can be used to prevent hyperketonemia in dairy cows. The objective of this study was to compare the beneficial effects of orally supplementing a mixture of GP on metabolic health indicators and milk production status of primiparous (PP) and multiparous (MP) dairy cows. Methods: Twenty-eight Holstein cows were blocked by expected date of parturition, previous lactation yield, and parity. The cows were randomly allocated to one of the four treatment groups (n = 7 cows/group) based on their parity and GP supplementation: i) PP cows fed basal diet only (PP-CON), ii) PP cows with oral supplementation of GP (PP-GP), iii) MP cows fed basal diet only (MP-CON), and iv) MP cows with oral supplementation of GP (MP-GP). Glucose precursor (glycoline liquid) was orally drenched (300 mL/d) in GP cows from 7 days prepartum through 7 days postpartum. Other than GP supplementation, all cows were fed similar pre- and postpartum basal diets. Results: In both pre- and postpartum periods, serum glucose concentration was increased, whereas β-hydroxybutyrate and free fatty acids were decreased in GP cows compared with the CON cows. Milk yield and milk components were statistically not different between GP and CON cows over the first 9 week of lactation. The yield of actual milk, energy-corrected milk, 63-days cumulative milk, colostrum yield, and calf birth weight remained higher in MP cows compared with PP cows. Conclusion: Oral drenching of GP around calving can be recommended to successfully improve the metabolic health and reduce the negative effects of hyperketonemia not only in MP but also in PP dairy cows.

• Analytical Science and Technology
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• v.22 no.4
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• pp.326-335
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• 2009

A specific analytical method able to identify and quantify traces of six glucocorticoids residues in eggs were developed. The extraction and clean-up parameters for simultaneous analysis were evaluated and HPLC and spectrometric conditions were also established. For determination of glucocorticoids, 5 g of egg was transferred into a test tube, adjusted pH 5.2 with acetate buffer and was $\beta$-glucuronidase/arylsulfatase from Helix pomatia added. The mixture was centrifuged and supernatant was extracted twice with 20 mL n-hexane. The extraction was performed with HLB cartridge using methanol, followed by clean-up with silica cartridge using methanol/ethyl acetate (4/6, v/v). The analytes were determined by HPLC/ESI-MS/MS operating in the negative ion mode. Validation studies with fortified egg samples for established method were performed. The result of method validation gave good efficiency, linearity, accuracy and precision. The correlation coefficients ($r^2$) of the calibration curves appeared to be higher than 0.99 in egg, indicating excellent linearity. LOD was ranged 0.09 to $0.17{\mu}g/kg$, and recoveries for most compounds were in the range of 55.7-69.8%. This method can be used to determine ${\mu}g/kg$ levels of glucocorticoids in eggs.

• Journal of Life Science
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• v.34 no.1
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• pp.59-67
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• 2024

LS-RUG-com preparation in a complex extract from mixture of three natural plants. Rubus crataegifolius/unriped, Gardenia jasminoides and Ulmus marcrocarpa that have been widely used in traditional functional health food. This study was conducted to establish the HPLC analysis methods that can be used to establish quantitative analysis of R. crataegifolius, G. jasminoides and U. macrocarpa for standardization of LS-RUG-com preparations. HPLC analysis methods for simultaneous determination of ellagic acid and geniposide and single determination of catechin-7-O-β-D-apiofuranoside were established for the quality control of natural plants complex (LS-RUG-com). Validation of HPLC analysis were performed by checking specificity, accuracy, precision, limit of detection and quantitation, and linearity following ICH (International Council for Harmonisation) guideline. As the result of quantitative analysis, the contents of ellagic acid, geniposide and catechin-7-O-β-D-apiofuranoside in each plant extracts were 11.2 mg/g (ellagic acid) and 72 mg/g (geniposide) and 10.2 mg/g (catechin-7-O-β-D-apiofuranoside). The contents of ellagic acid, geniposide and catechin-7-O-β-D-apiofuranoside in LS-RUG-com were 4.62~6.82 mg/g (ellagic acid), 19.2~28.8 mg/g (geniposide) and 1.36~2.04 mg/g (catechin-7-O-β-D-apiofuranoside) respectively.

• Journal of Life Science
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• v.20 no.5
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• pp.750-759
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• 2010

The bioactive materials (phenolic compounds, flavonoids, minerals, decursin and decursinol angelate) and biological activities (DPPH [$\alpha,\alpha$'-diphenyl-$\beta$-picrylhydrazyl] free radical scavenging capability, reducing power, and tyrosinase activity) in the extracts of leaf, stem mixture (AGLS), and root (AGR) from Angelica gigas Nakai were examined by using water, hot water and ethanol solvent. The highest extract yield (21.89%) was found in the water extract of AGR. The highest concentrations of phenolic compounds and flavonoids in the ethanol extracts of AGLS and AGR were 14.99% and 14.79%. Major minerals of AGLS and AGR were K, Mg, Fe, Na and Ca. Decursin and decursinol angelate were the major ingredients of Angelica gigas, detected at 18.71 and 18.89 min of retention time by HPLC analysis, respectively. The highest concentrations of decursin and decursinol angelate in the Angelica gigas ethanol extract were found in root ($41.7\;{\mu}g/g$) and leaf ($34.04\;{\mu}g/g$). The highest free radical scavenging activity was found in the hot water extracts of AGLS and AGR, and its activity was stronger in all extracts of AGLS than AGR. The highest reducing power was found in the ethanol extracts of AGLS and AGR and this was dependent on the sample concentration. The hot water extracts of AGLS and AGR revealed the highest inhibition activity on tyrosinase. Overall, these results may provide the basic data needed to understand the biological activities of bioactive materials derived from Angelica gigas.