• Journal of the Society of Cosmetic Scientists of Korea
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• v.26 no.1
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• pp.163-186
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• 2000

We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.3 no.1
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• pp.32-37
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• 2017

Cis-(1S,4S)-4-(3,4-dichlorophenyl)-1,2,3,4-tertrahydro-N-methyl-1-naphthalenamine (sertraline) hydrochloride from among selective serotonin reuptake inhibitors (SSRIs) is a treatment of major depression. For the differential diagnosis by metabolizing serotonin in a patient with neurological disorders, the radiolabeled $^{11}C$-sertraline was developed for non-invasive positron emission tomography in living brain and use the evaluation of new drug for SSRIs. We release the results of a fast and easy radiolabeling method applied a one-step loop method with $[^{11}C]CH_3OTf$ for routine clinical applications of $^{11}C$-sertraline. 1 mg of a precursor for $^{11}C$-sertraline in 0.1 mL DMF and $5{\mu}L$ of 1N NaOH, were injected into the loop of semi-prep high-performance liquid chromatography (HPLC). $[^{11}C]CH_3OTf$ was passed through the loop at room temperature (RT). The $^{11}C$-sertraline was separated by the semi-preparative HPLC. $^{11}C$-sertraline was eluted at 28.0 min was collected and evaluated by analytical HPLC and mass spectrometer. The total radiolabeling efficiency of $^{11}C$-sertraline was $30.7{\pm}8.7%$. The specific activity was $64.8{\pm}51.4GBq/{\mu}mol$. The radiochemical and chemical purities were higher than 99%. The mass spectrum of the product showed m/z peaks at 307.1 (M+1), indicating the mass of sertraline. By the one-step loop method with $[^{11}C]CH_3OTf$, $^{11}C$-sertraline could be quickly and easily prepared for clinical application.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.8
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• pp.1099-1104
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• 2017

Objective: This work was to find the correlation of alcohol dehydrogenase 1C (ADH1C) genotype with vitamin A reduction and carcass traits during the vitamin A restriction period. Methods: In study 1, 60 Korean native steers were fed a diet (890 IU/kg) with 8,000 IU and 0 IU of supplemental premix vitamin A/kg of dry matter (DM) for control and treatment group, respectively. The levels of serum vitamin A were analyzed through high preparative performance liquid chromatography, and the ADH1C genotype was analyzed based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP; 78.1% TT type, 21.9% TC type); however, CC type was not found. Then, the interaction between ADH1C and carcass traits on the vitamin A restriction was investigated in study 2. A total of 136 Korean native steers were fed a diet that included 930 IU/kg vitamin A of DM. Results: Serum vitamin A in treatment was reduced to 112.4 IU/dL in steers with TT type of ADH1C, while for steers with TC type the concentration of serum vitamin A was dropped to 79.5 IU/dL (p<0.1) in study 1. This showed that TC type had the potential to lower serum vitamin A concentration during vitamin A restriction compared to TT type. In study 2 we found that eye muscle area, marbling and carcass weight in Korean native steers with TC type were higher than in steers with TT type (p<0.05). Conclusion: The interaction between vitamin A restriction and TC type of ADH1C gene could have the potential of increasing the marbling in Korean native steers. These results indicated that steers with TC type of the ADH1C gene were more sensitive to the change of serum vitamin A than TT types. Furthermore, this finding has the potential to enable a higher marbling score under the condition of vitamin A restriction in Korean native steers.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.755-762
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• 2005

Biochemical amounts of RNA molecules can be synthesized in vitro, which is functionally equivalent or similar to those transcripts normally existing at extremely low levels in vivo. In this study we described a method for efficient preparation of pure T7 RNA polymerase from Escherichia coli strain BL21/pAR1219. The procedure, which used ammonium sulfate fractionation and preparative column chromatography on sephadex SP, was shown to be simple, rapid, and cost effective in comparison with other methods reported previously, Using the purified T7 RNA polymerase we were able to synthesize very long RNA transcript of 1.54 kb length, which is not feasible by conventional chemical synthesis. RNA molecule that was also synthesized by the purified T7 RNA polymerase, such as hammerhead ribozyme, retained its biochemical activity by cleaving the target RNA successfully in vitro. Thus, the procedure shown in this study can be useful to synthesize any length of RNA molecules in vitro in a simple and cost effective way for a variety of purposes.

• Korean Journal of Microbiology
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• v.28 no.2
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• pp.151-157
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• 1990

The maximum nitrogen fixation activity of the associative, microaerobic and acid tolerant bacteria, Azospirillum amazonense Kp1 was obtained with 0.2Kpa of $O_{2}$ and showed a reversible inhibition by the higher concentrations. Ammonium treatment caused a gradual inhibition of the activity up to 350mM. The nitrogenase systems were purified by gradient chromatography on DEAE-52 cellulose, heat treatment and preparative PAGE. The MoFe protein showed molecular weight of 210,000 including two nonidentical subunits with apparent molecular weights of 55,000 and 50,000 and an isoelectricpoint of 5.2 and contained 2, 24 and 28 atoms of Mo, Fe and acid labile S per molecule. The Fe protein revealed molecular weight of 66,000 including two types of subunits with molecular weights of 35,000 and 31,000 and an isoelectric point of 4.6, and contained 4 atoms of Fe and 6 atoms of S per molecule. The maximum specific nitrogenase activity attained 2,200 and 1,700nM $C_2H_4mg^{-1} min^{-1}$, respectively for MoFe and Fe proteins at pH7 and $35^{\circ}C$. The activity was lost after 10 and 30 days under the cold room ($4^{\circ}C$) condition for Fe and MoFe proteins, respectively.

• Journal of Applied Biological Chemistry
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• v.60 no.1
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• pp.49-54
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• 2017

In the course of screening tyrosinase inhibitory activity, EtOAc-soluble fraction of Coix lachryma-jobi var. mayuen Stapf. (Gramineae) roots showed significant inhibition. Further fractionation of the EtOAc-soluble fraction resulted in six compounds, which were identified as (+)-icariol $A_2$ (1), zhepiresionol (2), 4-hydroxybenzaldehyde (3), trans-${\rho}$-coumaric acid (4), N-(2-hydroxy-4-methoxyphenyl)-2-hydroxyacetamide (5), and coixol (6). The chemical structures of these compounds were identified on the basis of spectroscopic methods (MS, 1D and 2D NMR) and comparison with literature values. Compound 1 was first reported from this plant. Also, this is the first time that the isolation of compound 5 has been reported from nature source. Among the isolated compounds, compounds 4 and 6 showed enzyme inhibitory activity, with $IC_{50}$ values of 6.5 and $62.4{\mu}M$, respectively, in comparison with these of positive control, arbutin.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.27 no.1
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• pp.133-150
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• 2001

자유라디칼은 반응성이 커서 세포의 구성 물질인 단백질, 지질, 당, DNA등과 반응하여 세포 및 조직을 손상시킴으로서 노화를 가져온다. 이러한 자유라디칼들은 자외선, 일반 대사, 스트레스, 질병, 흡연 등에 의해 생성되고 특히, 활성 산소 종인 hydroxyl radical, superoxide radical, lipid peroxide radical등은 피부 노화를 발생시키는 원인이 된다. 따라서 본 연구에서는 한방 식물로부터 유해한 활성 산소종을 소거하는 활성을 가진 물질을 기 위해 1차 스크리닝 하였고, 이중 육두구, 관중, 초두구, 빈랑, 금은화, 포공영, 우롱차, 황금, 녹차 추출물이 효과가 높게 나타났으며, 그중 초두구와 빈랑 90% 메탄올 추출물에서 가장 우수한 소거 활성을 나타냄으로써 이 추출물에 대하여 자유라디칼 소거 활성에 대하여 여러 가지 생물학적 활성을 조사하였다. 초두구와 빈랑추출물에 대한 hydroxyl radical, superoxide radical, lipid peroxide radical을 소거하는 활성을 핵산, 클로로포름, 에틸아세테이트, 물층으로 분획하여 조사한 결과 에틸아세테이트층에서 가장 우수한 활성을 보였다. 에틸아세테이트 분획에 대한 lipid peroxide radical 소거에 있어서 빈랑 추출물이 $IC_{50}$/ 값 50 $\mu\textrm{g}$/ml, 초두구 추출물은 $IC_{50}$/ 값 59 $\mu\textrm{g}$/ml로서 기준물질로 사용되는 vitamic C (120 $\mu\textrm{g}$/ml)나 butylated hydroxyl toluene (80 $\mu\textrm{g}$/ml) 보다 더 우수한 소거 효과를 보여주었고, hydroxyl radical을 소거하는 능력은 25 $\mu\textrm{g}$/ml와 150 $\mu\textrm{g}$/ml로 hydroxyl radical의 소거 능이 좋은 vitamin C (180 $\mu\textrm{g}$/ml) 보다 뛰어난 소거 활성을 나타내었다. Superoxide radical을 소거하는 효과는 초두구 추출물의 $IC_{50}$/ 값 10 $\mu\textrm{g}$/ml, 빈랑 추출물이 15 $\mu\textrm{g}$/ml을 나타냈고, 이는 기준 물질인 vitamin C (35 $\mu\textrm{g}$/ml)보다 좋은 소거 활성을 보여주었으며, gallic acid 9 $\mu\textrm{g}$/ml과 유사한 효과를 나타내었다. 사람 섬유아세포를 배양하여 hydroxyl radical과 superoxide radical를 발생시킨 후 초두구와 빈랑 추출물의 세포 보호 효과를 실험한 결과 25 $\mu\textrm{g}$/ml의 농도로 처리하였을 때 각각 85% 이상의 우수한 세포 보호 효과를 나타내었다. 초두구로부터는 자유라디칼 소거 활성이 있는 물질을 분리하기 위하여 분획한 후 가장 높은 소거 활성을 보인 에틸아세테이트 층에 대하여 silica column chromatography, preparative TLC를 수행하였다. 초두구로부터 분리된 물질은 HPLC를 이용한 분리에서 phenol성 물질인 gallic acid와 동일한 retention time을 보여줌으로써 초두구로부터 분리된 물질은 gallic acid와 유사한 phenol성 물질이거나 그의 유도체일 것으로 추측된다. 따라서, 초두구와 빈랑 추출물은 피부 노화의 주요인이 되고 있는 lipid radical, hydroxyl radical, superoxide radical을 소거하는 활성이 뛰어나 자유라디칼에 의하여 발생되는 피부노화를 방지할수 있는 물질로서의 효과가 기대된다.가 기대된다.

• Korean Journal of Food Science and Technology
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• v.13 no.1
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• pp.57-66
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• 1981

An effective method for isolation of the major components of ginseng saponin such as $ginsenoside-Rb_{1},\;-Rb_2,$ -Rc, -Rd, -Re and $-Rg_1$, and the minor components such as ginsenoside-Rf, $-Rg_2,\;and-Rh_1$, was developed and reported in previous papers (J. Korean Agr. Chem. Soc., 23(4), 199 and 206(1980) The conditions and procedures used for isolation and identification for ginsenosides described in the previous papers were not sufficient enough for clean separation of minor components, $ginsenoside-Rh_1,\;and-Rh_2$. In this work, modifications in extraction method and in mobile phase for HPLC were attempted. It was found that application of ethyl acetate extraction at $60^{\circ}C$ for 3 hr on crude saponin resulted in a removal of diol group saponin from crude saponin which made it possible for using higher portion of acetonitrile in mobile phase. The mixed solvents of acetonitrile : water (92 : 8 and 94 : 6) gave excellent resolution of $ginsenoside-Rh_1\;and\;-Rh_2$.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.274-282
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• 2002

The ethanol extract and n-hexane fraction from Hypericum ascyron L. showed strong growth inhibition at 25 ppm on 5 strains of Listeria monocytogenes for 72 hr at $32^{\circ}C$. The purified substance, H2-5-2 fraction, was isolated by silica gel column and preparative thin layer chromatography from n-hexane fraction of Hypericum ascyron L. The H2-5-2 fraction showed a strong bacteriostatic activity on 5 strains of L. monocytogenes at 10 ppm in tryptic soy broth, and the viable cell was reduced 1 log cycle compared to initial cell number. The n-hexane fraction of Hypericum ascyron L. showed strong growth inhibition at 25 ppm on Bacillus cereus and Staphylococcus aureus, and at 50 ppm on Vibrio parahaemolyticus for 72 hr. The purified antimicrobial substance, the H2-5-2 fraction, was assumed as high unsaturated sterol by $^1H-NMR$ and $^{13}C-NMR$. On application test using minced Alaska pollack and ground beef, the n-hexane fraction of Hypericum ascyron L. at the level of 250 ppm was applied at $32^{\circ}C$ and $5^{\circ}C$. At $32^{\circ}C$ storage condition, the antimicrobial substances did not reduced L. monocytogenes ATCC 19113, meanwhile at $5^{\circ}C$ storage condition, L. monocytogenes ATCC 19113 was reduced in viable number.

• Korean Journal of Food Science and Technology
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• v.29 no.3
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• pp.417-426
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• 1997

Rice bran protein hydrolysates were prepared and some of their physicochemical properties were investigated to utilize rice bran as starting material for functional food ingredient. Rice bran proteins (RBP) were prepared from defatted rice bran by alkaline extraction and isoelectric precipitation. The enzyme for hydrolysis of RBP was selected through measuring relative activity by pH-drop method and comparing the degree of hydrolysis (DH) of hydrolysates. The enzymatic hydrolysates prepared by $Esperase^{\circledR}$ treatment were partitioned into two fractions by ultrafiltration(UF) with a 10 kDa molecular weight cut-off membrane. Each fraction was applied to a cholic acid-conjugated ${\omega}-aminohexyl$ Sepharose 4B column and the bile acid-binding components were obtained by eluting with deoxycholate. Gel permeation chromatography on a Sephadex G-50 column revealed that molecular weight of the bile acid-binding fraction of UF permeate was distributed in ranges of $2\;kDa{\sim}10\;kDa$ and $0.2\;kDa{\sim}0.6\;kDa$. Three peaks (R-1, R-2 and R-3) were obtained by prep-HPLC of bile acid-binding fraction of UF retentate and analyzed for total and free amino acid composition. The results showed that proline content of the bile-acid binding polypeptides and peptides was four times as much as that of rice bran protein and that the peak corresponding to higher average hydrophobicity had a higher free amino acid content. Average hydrophobicity slightly increased with enzymatic hydrolysis.