• Asian-Australasian Journal of Animal Sciences
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• 제17권5호
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• pp.712-718
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• 2004

This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

• 한국응용과학기술학회지
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• 제29권1호
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• pp.95-101
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• 2012

완전 생분해성 고분자 블렌드필름을 제조하기 위하여 치환도가 다른 두 셀룰로오스 아세테이트(CA)에 5 - 50%의 저분자량 폴리락타이드(PLA)를 블렌딩하였다. 이 때 사용된 각각의 고분자는 10% 메탄올/메틸렌클로라이드 혼합용제에 녹여서 점도가 같은 조건의 농도로 제조하였다. 각 조성의 블렌드필름의 표면 모폴로지와 열적 성질, 기계적 성질을 조사하였다. 화학적 구조는 적외선 분광법으로 확인하였으며, 전자현미경을 통한 표면 분석 결과 5% 이하의 폴리락타이드를 함유한 블렌드필름은 상분리가 거의 일어나지 않았으며 20% 이상이 함유된 경우 상분리가 매우 심각하였다. 또한 블렌드필름의 인장강도는 셀룰로오스 아세테이트의 함량이 늘어날수록 TAC/PLA의 경우 $820kg_f/cm^2$ 및 DAC/PLA의 경우 $600kg_f/cm^2$까지 향상시킬 수 있었다.

• 청정기술
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• 제12권4호
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• pp.198-204
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• 2006

생분해성 고분자인 폴리카프로락톤을 반응개시제를 첨가하여 반응압출법으로 개질시켰다. 개시제의 함량은 중량 백분율로 0.1, 0.3, 0.5 그리고 1.0% 이었고 이축압출기의 온도는 $130{\sim}180^{\circ}C$로 운전하였다. 개질된 폴리카프로락톤에 대한 열분석, 분자량 분석, 기계적 강도 측정, 유변학적 특성 및 생분해도 연구를 수행하였다. 개질된 폴리카프로락톤은 순수 폴리카프로락톤과 비교하여 여러 물성에서 차이를 보였다. 개시제 1% 개질 폴리카프로락톤은 약 20%의 결정화도 감소, 약 50%의 인장탄성률 상승을 보였고, 복합점도, 유변학적 특성인 저장탄성률 및 손실탄성률의 큰 증가를 보여주었다. 0.1%의 가장 적은 개시제로 개질된 폴리카프로락톤의 생분해도는 순수 폴리카프로락톤과 비슷한 생분해도를 보였으나, 나머지는 생분해도가 증가하였다.

• Elastomers and Composites
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• 제44권2호
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• pp.175-181
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• 2009

편광필름용 점착제를 합성하기 위해 아크릴 모노머를 사용하여 용액중합을 수행하였다. 아크릴 모노머는 2-ethylhexyl acrylate, butyl acrylate, acrylic acid를 사용하였으며, 개시제는 benzoyl peroxide, 용매는 ethyl acetate가 사용되었다. 모노머의 비율은 점착제의 유리전이온도 $-40^{\circ}C$에 맞추어 2-ethylhexyl acrylate: butyl acrylate: acrylic acid = 25:50:3.6 이였다. 개시제의 첨가량은 점착제의 젖음성과 초기점착력을 고려하여 모노머 대비 0.09%로 결정하였다. 모노머와 용매의 비율은 젖음성과 투과율을 고려하여 1:1.7로 하였다. 점착제 필름의 투과율은 점도가 낮을수록, 분자량이 작을수록, 분자들 간의 얽힘이 적어져서 굴절율이 낮아지고 투과율이 증가함을 알 수 있었다. 사용가능 기간의 측정에서 모노머와 용매의 비율이 1:1.7일 경우 점도가 200분이 지나도 커다란 변화가 없어 저장안정성이 좋음을 확인할 수 있었다.

• 한국약용작물학회지
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• 제3권1호
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• pp.61-65
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• 1995

한국에서 재배되고 있는 시호(Bupleurum falcatum L.)의 유전적 계통의 확립을 위해 시호(Bf), 참시호(BfC), 대전 삼도시호(BfJ) 및 수원 삼도시호(BfS)등 4계통을 대상으로 핵형분석을 통해 염색체의 다형현상을 비교하고, SDS PAGE를 이용하여 저장단백질의 양상을 분석하였다. 체세포 염색체 수는 3계통(Bf, BfC, BfJ)에서 2n=20, 1 계통(BfS)에서 2n=26으로 구분되었다. 염색체 1번은 2n=20 계통의 경우 차중부 염색체였으나, 계통 2n=26에서는 중부 염색체로 관찰되어 차이를 보였다. 계통 2n=20에서 2번, 3번 및 5번염색체가 다형현상을 보였으며, 한국에서 재배되고 있는 계통의 핵형은 일본에서 도입되어 재배되고 있는 대전 삼도시호 및 수원 삼도시호와 차이를 보였다. 이러한 결과는 세포유전학적으로 다른 2가지 이상의 계통이 한국에서 재배되고 있음을 보여주는 것이다. 종자에서 추출한 저장 단백질은 45KD 이상의 단백질에서 수적 차이를 보였으며, 2n=20의 계통에서 3개, 2n=26 계통에서 2개의 특이한 밴드가 관찰되었다.

• Journal of Pharmaceutical Investigation
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• 제30권3호
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• pp.159-166
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• 2000

Kunitz-type soybean trypsin inhibitor (SBTI) and chondroitin sulfate (A, and C type) were conjugated using sodium periodate method. And the physicochemical, pharmacokinetic properties and immunogenecity of the conjugates (Chon-A-SBTI or Chon-C-SBTI) were characterized. We expected the conjugation using chondroitin sulfate to reduce the immunogenecity and to improve the pharmacological effect. As the results, the mean molecular weight of the conjugate highly increased. After I.V. injection of the radiolabeled conjugates or native SBTI into mice, it was found that native SBTI showed rapid elimination from plasma, whereas Chon-A-SBTI and Chon-C-SBTI were slowly eliminated. Organ distribution of the two agents at 30 min after I.V. injection was different : Chon-A-SBTI or Chon-C-SBTI accumulated to a large extent in the liver (13% in Chon-A-SBTI and 16% in Chon-C-SBTI), whereas native SBTI was taken up more rapidly by the kidney (107% dose/g of tissue) and excreated into the urine (26%). In addition we evaluated the therapeutic value of the conjugates by using the sublethal septic shock model caused by pseudomonal elastase and tested the immunogenecity by passive cutaneous anaphylaxis shock (PCA). The conjugates were more effective than native SBTI against pseudomonal elastase induced septic shock in guinea pig. In case of the conjugates, the pharmacological and therapeutic effect lasted over 3 hours long. In immunogenecity test, both of the conjugates showed the reduction of their immunogenecity, especially Chon-A-SBTI looked most effective.

• Journal of Periodontal and Implant Science
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• 제38권1호
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• pp.41-50
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• 2008

Purpose: Bone morphogenetic protein-2(BMP-2) has been shown to possess significant osteoinducitve potential. There have been attempts to overcome a limitation of mass production, and economical efficiency of BMP. The aim of this study was to produce recombinant human BMP-2(rhBMP-2) from E. coli in a large scale and evaluate its biological activity. Materials and Methods: The E.coli strain BL21(DE3) was used as a host for rhBMP-2 production. Dimerized rhBMP-2 was purified by affinity chromatography using Heparin column. To determine the physicochemical properties of the rhBMP-2 expressed in E. coli, we examined the HPLC profile and performed Western blot analysis. The effect of the purified rhBMP-2 dimer on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and representing morphological change using C2C12 cell. Results: E. coli was genetically engineered to produce rhBMP-2 in a non-active aggregated form. We have established a method which involves refolding and purifying a folded rhBMP-2 dimer from non-active aggregates. The purified rhBMP-2 homodimer was characterized by SDS-PAGE as molecular weight of about 28kDa and eluted at 34% acetonitrile, 13.27 min(retention time) in the HPLC profile and detected at Western blot. The purified rhBMP-2 dimer stimulated ALP activity and induced the transformation from myogenic differentiation to osteogenic differentiation. Conclusion: rhBMP-2 was produced in E. coli using genetic engineering. The purified rhBMP-2 dimer stimulated ALP activity and induced the osteogenic differentiation of C2C12 cells.

• Genomics & Informatics
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• 제6권3호
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• pp.142-146
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• 2008

In order to understand the protein functions that are related to disease, it is important to detect the correlation between amino acid mutations and disease. Many mutation studies about disease-related proteins have been carried out through molecular biology techniques, such as vector design, protein engineering, and protein crystallization. However, experimental protein mutation studies are time-consuming, be it in vivo or in vitro. We therefore performed a bioinformatic analysis of known disease-related mutations and their protein structure changes in order to analyze the correlation between mutation and disease. For this study, we selected 111 diseases that were related to 175 proteins from the PDB database and 710 mutations that were found in the protein structures. The mutations were acquired from the Human Gene Mutation Database (HGMD). We selected point mutations, excluding only insertions or deletions, for detecting structural changes. To detect a structural change by mutation, we analyzed not only the structural properties (distance of pocket and mutation, pocket size, surface size, and stability), but also the physico-chemical properties (weight, instability, isoelectric point (IEP), and GRAVY score) for the 710 mutations. We detected that the distance between the pocket and disease-related mutation lay within $20\;{\AA}$ (98.5%, 700 proteins). We found that there was no significant correlation between structural stability and disease-causing mutations or between hydrophobicity changes and critical mutations. For large-scale mutational analysis of disease-causing mutations, our bioinformatics approach, using 710 structural mutations, called "Structural Mutatomics," can help researchers to detect disease-specific mutations and to understand the biological functions of disease-related proteins.

• 약학회지
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• 제57권2호
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• pp.125-131
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• 2013

Biologically active peptides, including growth factors and cytokines, participate in various biological processes in human skin. They could provide a great advantage of maintaining healthy skin. Many peptide growth factors like epidermal growth factor (EGF) and human growth hormone (hGH) have been used in cosmetic formulations. The delivery of peptide growth factors across the Stratum corneum, however, seems not sufficient because of their physical properties such as high molecular weight and hydrophilicity. So increasing the penetration of growth factors of interest into skin would be a major concern for ensuring their maximum biological efficacy. In this study, we have identified several skin penetration-enhancing peptides which facilitate delivery of growth factors, when fused at N-terminus of the target protein, into skin. For efficient and rapid screening, we constructed a skin-penetrating assay system using Franz cell and porcine skin. Next, we carried out phage display screening using M-13 bacteriophage with random 12 -amino acid library on its coat protein P3 on that system. After several selection rounds, peptide sequences facilitate the penetration of phages through the porcine skin were identified from a large population of phages. We found that phages with the most potent peptide (S3-2, NGSLNTHLAPIL) could penetrate the porcine skin eight times more than those with control peptide (12 mino acids scrambled peptide). Furthermore, growth factors conjugated with S3-2 peptide penetrate porcine skin three to five times efficiently than non-conjugated growth factors. In conclusion, our data shows that the skin penetration-enhancing peptide we have characterized could increase the delivery of growth factors and is useful for cosmeceutical application.

• 대한약침학회지
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• 제3권2호
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• pp.1-25
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• 2000

We were trying to study the validity of Puffer fish's poison(Tetrodotoxin- TTX) to make a traditional Korean Medical treatment. The following conclusions were made after literary studies. 1. The first record of the puffer fish dates back 2000 years ago in the Chinese text Book of Mountain and Sea and other texts from the similar period. 2. Puffer fish's poison IS known as tetrodotoxin which is an amino perhydroquinazoline compound. It has a chemical formula of $C_{11}H_{17}N_3O_8$ in the hemiacetal structure and has the molecular weight of 319. 3. Tetrodotoxin (TTX) plays a role as potent neurotransmitter blocker by blocking the $Na^+$ -gate channel which hinders the influx of $Na^+$ ion into the cell. 4. Symptoms of the puffer fish poisoning ranges from blunted sense in the lips and tongue, occasional vomiting in the first degree to sudden descending of the blood pressure, apnea, and other critical conditions in the fourth degree. Intoxication of the puffer fish poison progresses at a rapid pace as death may occur after an hour and half up to eight hours in maximum. Typical death occurs after four to six hours. 5. Ways to treat the puffer fish poisoning include gastric irrigation, induce vomiting, purgation, intravenous fluid injection, and correcting electrolytic imbalance and acidosis. In cases of dyspnea, apply oxygen inhalation and conduct artificial respiration. 6. Tetrodotoxin (TTX) may be applied in treating brain disorders, ocular pain, excess pain in the large intestine and ileum, and relieving tension of the skeletal museles, neuralgia, rheumatism, arthritis, and etc. 7. In terms of Oriental medicine, the puffer fish poison has characteristics of sweet, warm, and poisonous. It's known efficacies are to tonify weakness, dispel damp, benefit the lower back, relieve hemorrhoid, kills parasites, remove edema, and so forth. And the puffer fish eggs processed with ginger are said to be effective against tuberculosis and lung cancer, thus, it's validity must be investigated and further research should be followed.