• 대한예방한의학회지
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• 제17권3호
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• pp.165-176
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• 2013

Objective : The purpose of this study was to establish the simultaneous analysis for six compounds in Gumiganghwal-tang (GMGHT, Jiuweiqianghuo-tang) and to investigate the anti-atherosclerotic effects of GMGHT in vitro. Methods : The column for separation of six compounds was used Luna $C_{18}$ column and maintained at $40^{\circ}C$. The mobile phase for gradient elution consisted of two solvent systems, 1.0% acetic acid in water and 1.0% acetic acid in acetonitrile. The analysis was carried out at a flow rate of 1.0 mL/min with pothodiode array (PDA) detection at 254, 280, and 320 nm. The injection volume was 10 ${\mu}L$. The antioxidant activities of GMGHT were evaluated by measuring free radical scavenging activities on 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH). The inhibitory effects on low-density lipoprotein (LDL) oxidation were evaluated by the formation of thiobarbituric acid relative substances (TBARS), relative electrophoretic mobility (REM), and fragmentation of apolipoprotein B (ApoB)-100. Results : Calibration curves were acquired with $r^2{\geq}0.9998$. The contents of liquiritin, ferulic acid, baicalin, baicalein, glycyrrhizin, and wogonin in GMGHT were 1.784, 1.693, 37.899, 0.258, 1.869, and 0.034 mg/g, respectively. The GMGHT showed the radical scavenging activity in a dose-dependent manner. The concentration required for 50% reduction ($RC_{50}$) against ABTS and DPPH radicals were 72.51 ${\mu}g/mL$ and 128.49 ${\mu}g/mL$. Furthermore, GMGHT reduced the oxidation properties of LDL induced by $CuSO_4$. Conclusion : HPLC-PDA is considered as an available and convenient method for quality control and standardization of GMGH and GMGHT has potentials on anti-atherosclerosis by anti-oxidative effect and suppressive effect on LDL oxidation.

• Bulletin of the Korean Chemical Society
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• 제33권3호
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• pp.790-794
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• 2012

Trace metal ions such as $Cd^{2+}$, $Ni^{2+}$, and $Zn^{2+}$ in a highly saline sample were subjected to on-line complexation with 4-(2-thiazolylazo) resorcinol (TAR) dissolved in a background electrolyte (BGE) under transient isotachophoresis (TITP) conditions. A long plug of the saline sample, containing the trace metal ions but devoid of TAR, was injected into a coated capillary filled with a BGE composed of 150 mM 2-(cyclohexylamino) ethanesulfonic acid (CHES) and 110 mM triethylamine (TEA) at pH 9.7. Since the electrophoretic mobility of TAR fell between the mobilities of the anionic leading electrolyte ($Cl^-$ in the sample) and the anionic terminating background electrolyte ($CHES^-$), a highly concentrated zone of TAR from the BGE was formed at the rear of the sample matrix and then the metal cations toward the cathode were swept by isotachophoretically assisted on-line complexation (IAOC) between the metal ions and the isotachophoretically stacked TAR. As a result, anionic metal-TAR complexes were formed efficiently, which satisfy the TITP conditions between $Cl^-$ and $CHES^-$. The enrichment factors of metal ions including $Cd^{2+}$ were up to 780-fold compared to a conventional CZE mode using absorbance detection. The detection limits were 17 nM, 15 nM, and 27 nM for $Ni^{2+}$, $Zn^{2+}$, and $Cd^{2+}$ in a 250 mM NaCl matrix, respectively. Our method was successfully applied to the analysis of urine samples without desalting.

• Molecules and Cells
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• 제27권1호
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• pp.113-118
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• 2009

In this study, we have shown the transcriptional regulation of the human GD3 synthase (hST8Sia I) induced by valproic acid (VPA) in human neuroblastoma SK-N-BE(2)-C cells. To elucidate the mechanism underlying the regulation of hST8Sia I gene expression in VPA-stimulated SK-N-BE(2)-C cells, we characterized the promoter region of the hST8Sia I gene. Functional analysis of the 5'-flanking region of the hST8Sia I gene by the transient expression method showed that the -1146 to -646 region, which contains putative binding sites for transcription factors c-Ets-1, CREB, AP-1 and NF-${\kappa}B$, functions as the VPA-inducible promoter of hST8Sia I in SK-N-BE(2)-C cells. Site-directed mutagenesis and electrophoretic mobility shift assay indicated that the NF-${\kappa}B$ binding site at -731 to -722 was crucial for the VPA-induced expression of hST8Sia I in SK-N-BE(2)-C cells. In addition, the transcriptional activity of hST8Sia I induced by VPA in SK-N-BE(2)-C cells was strongly inhibited by SP600125, which is a c-Jun N-terminal kinase (JNK) inhibitor, and $G{\ddot{O}}6976$, which is a protein kinase C (PKC) inhibitor, as determined by RT-PCR (reverse transcription-polymerase chain reaction) and luciferase assays. These results suggest that VPA markedly modulated transcriptional regulation of hST8Sia I gene expression through PKC/JNK signal pathways in SK-N-BE(2)-C cells.

• 한국식품과학회지
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• 제20권1호
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• pp.90-94
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• 1988

본 연구는 효모를 동정하는 방법으로서 각 효모의 단백질 조성 차이와 이들 단백질의 면역학적 특성에 의한 새로운 동정법을 개발하고자 하였다. S. cerevisiae, C. utilis, C. tropicalis, K. fragilis 의 4가지 효모를 배양하여 세포를 파괴시킨 다음 가용성 단백질과 막 단백질을 분리 추출하였다. 4종 효모의 가용성 단백질과 막 단백질의 조성은 SDS-polyacrylamide gel electrophoresis를 실시 비교하였다. S. cerevisiae와 C. utilis 는 전기영동 pattern상 유사하였고 이들은 쉽게 C. tropicalis, K. fragilis 로부터 구별이 가능하였다. 또한 4종 효모의 가용성 단백질과 막 단백질을 토끼에게 주사하여 각각에 대응하는 항체를 만든 후 Ouchterlony double immunodiffusion을 실시하여 형성된 precipitin line에 의한 효모의 동정을 수행하였다. 면역학적으로도 S. cerevisiae와 C. utillis의 유사성이 증명되었고 이들은 C. tropicalis, K. fragilis 와 상이함이 관찰되었다.

• 한국식품과학회지
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• 제20권1호
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• pp.34-39
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• 1988

여러가지 육가공품(肉加工品)중 특정 단백질 원료의 첨가여부를 판정하여 변조식품(變造食品)을 검출하는 한 방법으로서, 각종 육류단백질, non-meat protein, 어육(魚肉)가공품을 대상으로 disc SDS-Poly acrylamide gel electrophoresis의 사용 가능성을 실험하였다. Total protein fraction에 대한 전기영동 결과 복잡하고 많은 band를 보여 각 시료에 고유한 특성을 찾아보기 어려웠다. Low salt-soluble protein fraction에서는 total protein fraction 에서 보다 band 수가 상당히 감소함을 보였고 각 단백질 원료에 대하여 보다 고유한 band pattern을 나타내었다. Acetone-insoluble protein fraction 에서는 non-meat protein의 경우 육류단백질과 상당히 다른 경향을 나타내었고. 소세지 원료의 가열처리에 의하여 단백질의 band수와 양이 감소하였다. 따라서 적당한 단백질 추출조건(抽出條件)을 설정하여 전기영동을 실시하면, 특정(特定) 단백질을 첨가한 변조식품의 검출이 가능해질 것으로 생각된다.

• Journal of Pharmaceutical Investigation
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• 제35권1호
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• pp.17-23
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• 2005

Polyethylenimine (PEI) has been used as cationic polymers for efficient gene transfer without the need for endosomolytic agents. Various kinds of PEIs with different molecular weight were tested in order to investigate the effects of the molecular weight of PEI on the transfection efficiency and cell cytotoxicity. The ${\beta}-galactosidase$ expression $(pCMV-{\beta}-gal)$ plasmid was used as a model DNA. Complex formation between PEI and pDNA was assessed by 1% agarose gel electrophoresis method. Particle size and zeta-potential of complexes were determined by electrophoretic light scattering spectrometer. In vitro transfection efficiency was assayed by measuring ${\beta}-galactosidase$ activity. Cell cytotoxicity was determined by MTT assay. Particle sizes of the complexes became smaller on increasing molecular weights of PEI and N/P ratios. Surface potential of complexes was increased as the molecular weight of PEI increased. Transfection efficiency of $pCMV-{\beta}-ga1$ on the HEK 293 cells was greatest with PEI 25 K system but having the lowest cell viability. PEI with high molecular weight showed higher transfection efficiency and cell viability than PEI with low molecular weight.

• 한국수산과학회지
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• 제21권2호
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• pp.85-96
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• 1988

Purification and some properties of alkaline proteinases in the pyloric caeca of skipjack, Katsuwonus vagans, were investigated. Four alkaline proteinases, temporarily designated proteinases I, II, III and IV, were identified from the tissue extract of the pyloric caeca by ammonium sulfate fractionation, DEAE-Sephadex A-50 chromatography, and Sephadex G-100 and G-200 gel filtration. Result of disc-polyacrylamide gel electrophoretic analysis showed that the purified proteinases II and III were homogenous with the yields of $1.5\%\;and\;1.2\%$, and those specific activities were increased to 33 to 37 fold over that of the crude enzyme solution, respectively. Molecular weight of the proteinases II and III determined by sephadex G-100 gel filtration were 28,500 and 24,200, respectively. The optimum conditions for the caseinolytic activity of the two enzymes were pH 9.6 and $48^{\circ}C$. The reaction rates of the two alkaline proteinases were constant to the reaction time to 80 min in the reaction mixture of $3.4{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The Km values against casein substrate determined by the method of Lineweaver-Burk were $0.56\%$ for proteinase II and $0.30\%$ for proteinase II. The proteinases II and III were inactivated under the presence of $Ag^+,\;Hg^{2+},\;Ni{2+},\;Fe^{2+},\;and\;Cu^{2+}$, and but activated by $Mn^{2+}\;and\;Ca^{2+}$ and markedly inhibited by the soybean trypsin inhibitor and N-p-toluenesulfonyl-L-lysine chloromethyl ketone. Therefore, the proteinases II and III were found to be a group of serine proteases and assured to be trypsin-like proteinases.

• Journal of Ginseng Research
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• 제22권3호
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• pp.168-172
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• 1998

Systematic electrophoretic analysis of alcohol-soluble proteins and salt-soluble proteins of 247 Panax ginseng (P.g) and Panax quinquefolium (P.q) germplasms seed was carried out on an improved lactate-polyacrylamide gel electrophoresis, a method with high resolving power, good reproducibility and stability. The electrophoregrams of proteins, according to their migration rate, were classified into four groups such as ${\alpha}$, ${\beta}$, ${\gamma}$ and $\omega$ for the alcohol-soluble proteins and three such as I, II and III for the salt-soluble ones. Panax ginseng or Panax quinquefolium had their own unique band pattern distinguishable from each other, regarding as their specific "fingerprint". In this study, 3 of 168 (1.8%) P.g germplasms and 1 of 79 (1.3%) P.q germplasms had their own unique band pattern, showing that P.g and P.q germplasms have poor genetic diversity in species. The band patterns of dry seed and stratified seed (embryo rate=60%) were basically the same. The band number of the F, hybrid of p.gx p.q was exactly equivalent to the number of the common bands plus the specific bands of the two parents, indicating that the difference of band patterns was a genetic trait con- trolled by the nuclear genes. The electrophoregram of F1 of P.g x P.q could be predicted by that of the two parents and the band pattern of the F1 hybrids could be demnonstrated by that of the mixed seed extract from the two parents.

• Asian-Australasian Journal of Animal Sciences
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• 제28권6호
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• pp.788-795
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• 2015

Two dimensional-fluorescence difference gel electrophoresis (2D DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. The purpose of this study was to investigate bovine pregnancy-specific proteins in the proteome between bovine pregnant and non-pregnant serum using DIGE technique. Serums of 2 pregnant Holstein dairy cattle at day 21 after artificial insemination and those of 2 non-pregnant were used in this study. The pre-electrophoretic labeling of pregnant and non-pregnant serum proteins were mixed with Cy3 and Cy5 fluorescent dyes, respectively, and an internal standard was labeled with Cy2. Labeled proteins with Cy2, Cy3, and Cy5 were separated together in a single gel, and then were detected by fluorescence image analyzer. The 2D DIGE method using fluorescence CyDye DIGE flour had higher sensitivity than conventional 2D gel electrophoresis, and showed reproducible results. Approximately 1,500 protein spots were detected by 2D DIGE. Several proteins showed a more than 1.5-fold up and down regulation between non-pregnant and pregnant serum proteins. The differentially expressed proteins were identified by MALDI-TOF mass spectrometer. A total 16 protein spots were detected to regulate differentially in the pregnant serum, among which 7 spots were up-regulated proteins such as conglutinin precursor, modified bovine fibrinogen and IgG1, and 6 spots were down-regulated proteins such as hemoglobin, complement component 3, bovine fibrinogen and IgG2a three spots were not identified. The identified proteins demonstrate that early pregnant bovine serum may have several pregnancy-specific proteins, and these could be a valuable information for the development of pregnancy-diagnostic markers in early pregnancy bovine serum.

• 생명과학회지
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• 제14권1호
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• pp.115-120
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• 2004

민자주방망이버섯은 우수한 식용버섯으로 세계적으로 분포하고 있는 종이다. 민자주방망이버섯과 5가지 송이과버섯 종들의 유전적인 변이와 분류학적 관계를 RAPD에 의해 분석 하였다. RAPD에 15가지 random primer를 사용하였다. 버섯들의 유전적 거리는 UPCMA를 사용하여 측정하였고 계통유전학적 유연관계는 neighnor-joining (NJ) 방법을 사용하여 분석하였다. PCR에 의하여 증폭된 RAPD 절편들은 100bp에서 1600 bp이었다. Nei-Li의 유전적 거리는 228개의 DNA밴드를 사용하여 계산하였고 이를 바탕으로 계통유전학적 계통수를 만들었다. 민자주방망이버섯, 자주방망이버섯 아재비, 가랑잎애기버섯, 밀버섯, 만가닥버섯, 졸각버섯의 유전적 변이 는 각각 0∼21.3%, 21.2∼28.0%, 15.4∼23.0%, 14.0∼21.8%, 16,5∼34.6%, 12.4∼27.4%이였다. CI (Consistency index), RI (Retention index), HI (Homoplasy inedx)의 값은 각각 0.5217, 0.5769, 0.5156이었다. 또한 NJ tree로 두 그룹을 만들 수 있었다. 유전적 거리는 민자주방망이버섯과 자주방망이버섯아재비보다 민자주방망이버섯과 밀버섯이 더 가까웠다.