• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권6호
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• pp.467-473
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• 2007

이 실험은 이식재로 Deproteinated Bovine Bone Powder (DBBP)를 사용하였을 때와 골 이식재를 사용하지 않고 단순히 Absorbable Gelatin Sponge(AGS)만을 사용하였을 때의 BMP4와 BMP6의 상대적인 발현량을 real-time RT-PCR을 이용하여 비교해 보기 위한 것으로 다음과 같은 결과를 얻었다. 1. BMP4의 경우 처음 1일째와 3일째의 경우 DBBP군과 AGS군 모두 일정하게 증가하였으나 5일째 AGS군에서 감소하다가 7일째 다시 증가하였으며 9일째 다시 감소하였다. DBBP군에서는 7일까지 계속하여 증가하다가 9일째 감소하였다. DBBP군이 AGS군에 비해 발현의 양이 많은 경우가 많았지만 유의성은 없었다 (p>0.05). 2 BMP6의 경우 처음 1일째와 3일째의 경우 DBBP군과 AGS군 모두 일정하게 증가하였으나 5일째 AGS군에서 감소하다가 7일째 다시 증가하였으며 9일째 다시 감소하였다. DBBP군에서는 7일까지 계속하여 증가하다가 9일째 감소하였다. AGS군이 DBBP군에 비해 발현의 양이 많은 경우가 많았지만 유의성은 없었다 (p>0.05). 3. 두 군간에 동일시기에 BMP발현이 유의할 만한 차이를 보이지 않는 것은 DBBP와 AGS 모두 space retainer로서 작용을 하여 혈병중의 BMP발현의 양상이 비슷하기 때문으로 여겨진다. 4. 따라서 DBBP가 AGS에 비해 초기 골재생에 크게 유리한 점은 없는 것으로 여겨지며 초기 골 형성에서 BMP의 발현은 이식재의 종류가 아니라 물리적인 carrier로서의 작용이 더 중요한 것으로 여겨진다.

• Reproductive and Developmental Biology
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• 제30권1호
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• pp.47-52
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• 2006

본 연구는 단위발생유래 생쥐 배아줄기세포(P-mES)지가 체외수정유래 생쥐 배아줄기세포 (mES)와 마찬가지로 기능성 심근세포로 체외 분화되는지를 조사하였다. 각 세포주 P-mES04와 MES03를 4일간 부유 배양하여 배아체 (EB)를 형성한 다음 4일간 DMSO를 추가적으로 처리한 뒤 젤라틴이 코팅된 배양접시에 부착시켰다(4-/4+). P-mES04와 mES03으로부터 수축성 심근세포 생성 여부를 30일간 관찰한 결과, 각각 13일(69.83%)과 22일 (61.3%)에 누적 형성율이 가장 높았다. 면역 세포화학염색 결과, 수축성을 나타내는 P-mES04 세포는 수축성 mES03 세포에서와 같이 근육 특이적인 anti-sarcomeric a-actinin 항체와 심근 특이적인 anti-cardiac troponin I 항체에 염색되는 것을 확인하였다. 또한 RT-PCR 결과, 수축성을 나타내는 P-mES04 세포는 심근특이적인 L-type calcium channel, a1C, cardiac myosin heavy chain a, cardiac muscle heavy polypeptide $7{\beta}$, GATA binding protein 4와 atrial natriuretic factor는 발현하나, 골격근 특이적인 L-type calcium channel, a1S는 발현하지 않아 웅성 성체의 심장세포와 유사한 양상을 보였다. 본 연구의 결과는 단위발생 유래 생쥐 배아 줄기세포를 배아줄기세포의 연구의 대체제로 이용할 수 있음을 보여준다.

• 자원리싸이클링
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• 제26권6호
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• pp.84-90
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• 2017

저품위 동 함유 슬러지로부터 회수된 동 조금속을 황산구리 전해액에서 전해정련을 수행하여 고순도 동을 회수하고자 하였다. 유기첨가제 종류 및 농도에 따른 전해정련동 표면 형상 및 조도를 분석하였을 때, gelatin계 유기첨가제 5 ppm 및 thiol계 유기첨가제 5~10 ppm 혼합 조건에서 가장 우수한 표면 형상을 얻을 수 있었다. 건식환원공정을 통해 회수한 철(Fe), 니켈(Ni), 코발트(Co) 및 주석(Sn) 등의 불순물이 함유된 각 86.635, 94.969 및 99.917 wt.% 순도의 동 조금속을 사용하여 전해정련하였고, 순도 3N~4N급 전해정련동을 얻을 수 있었다. 동 조금속, 전해액, 전해정련동의 불순 원소 농도 및 동 순도 등의 분석을 통해, 순도 99.99 wt.% 이상 전해정련동을 회수할 수 있는 전해정련 공정시간 및 동 조금속 순도를 도출하였다.

• Journal of Microbiology and Biotechnology
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• 제16권12호
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• pp.1904-1911
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• 2006

The cell-free extracts of 250 yeasts were screened for their in vitro anti-angiogenic activity, to develop a new cancer metastasis inhibitor. Saccharomyces cerevisiae K-7 was selected as the producer of the anti-angiogenic agent, because it had the highest anti-angiogenic activity. The anti-angiogenic agent was produced maximally from hydrolysates of Saccharomyces cerevisiae K-7, when the yeast was cultured in yeast extract-peptone-dextrose medium at 30$^{\circ}C$ for 24 h, and cell-free extracts were than digested with pepsin for 4 h at 37$^{\circ}C$. The anti-angiogenic agent was further purified by ultrafiltration, Sephadex G-25 gel permeation chromatography and reverse-phase HPLC, and the anti-angiogenic activity of the final purified preparation was 72.7% at 10 $\mu$M/egg. The purified anti-angiogenic agent was found to originate from the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) molecule of Saccharomyces cerevisiae K-7, and its peptide sequence was Val-Ser-Trp-Tyr-Asp-Asn-Glu-Tyr-Gly-Tyr-Ser-Thr-Arg-Val-Val-Asp. In the MTT assay, the shape of the HT-l 080 cell was clearly changed to a circular type at 0.2 mM purified anti-angiogenic agent. This result indicated that the growth of the HT-I080 cell was significantly inhibited at 0.2 mM of the purified anti-angiogenic agent. The MMP activity of the treated HT-l080 cells was not affected, evidenced by the gelatin zymography, indicating that the anti-angiogenic mechanism of the purified anti-angiogenic agent is not mediated through MMP activity.

• Journal of Nutrition and Health
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• 제36권3호
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• pp.280-286
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• 2003

Conjugated linoleic acid (CLA) consists of several geometric isomers of linoleic acid. CLA is found in foods derived from ruminants and exhibits strong anticarcinogenic effects in a variety of animal models. Matrix metalloproteinases (MMPs) play a key role in cancer progression. Specifically, MMP-2 and -9, which hydrolyze the basal membrane type IV collagen, are involved in the initial breakdown of collagen and basement membrane components during tumor growth and invasion. However, the effects of CLA on cancer cell motility and MMP expression and activity are not currently well known. Therefore, the present study examined whether CLA reduces the activity of MMP and cell motility in SW480 and SW620 cells, the human colon cancer cell lines. Gelatin zymography and Western blot analysis revealed that phorbol 12-myristate 13-acetate (PMA) induced the activity and protein expression of Mr 92,000 MMP-9 in both cell lines. To examine whether CLA inhibits the MMP activity, cells were incubated with 100 ngfmL PMA in the presence of various concentrations of CLA. PMA-induced MMP-9 activity was decreased by 20 $\mu$ M CLA in SW480 cells, and by 10 $\mu$ M and 20 $\mu$ M CLA in SW620 cells. Results from the Hoyden chamber assay showed that cell motility was increased by PMA and that PMA-induced cell motility was significantly decreased by 20 $\mu$ M CLA in SW480 cells. These results indicate that CLA may reduce the motility and MMP activity in human colon cancer cells.

• Nutritional Sciences
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• 제7권3호
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• pp.144-150
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• 2004

We have investigated the effect of calcium spirulan(Ca-SP) isolated from a blue-green alga Spirulina platensis, which is a sulfated polysaccharide chelating calcium and mainly composed of rhamnose and fructose, on invasion of both B16- BL6 melanoma cells, Colon 26 carcinoma and HT-1080 fibrosarcoma cells through reconstituted basement membrane (Matrigel). Ca-SP significantly inhibited the invasion of these tumor cells through Matrigel/fibronectin-coated filters in a concentration-dependent manner. Ca-SP also inhibited the haptotactic migration of tumor cells to laminin, but it had no inhibitory effect on tumor cell migration to fibronectin-coated filters. Ca-SP prevented the adhesion of B16-BL6 cells to Matrigel- and laminin-substrates but did not affect the adhesion to fibronectin. The pretreatment of tumor cells with Ca-SP inhibited the adhesion to laminin in a concentration-dependent fashion, while the pretreatment of laminin-substrates did not. Ca-SP had no effect on the production and activation of type IV collagenase in gelatin zymography. In contraset, Ca-SP significantly inhibited degradation of heparan sulfate by purified heparanase. The experimental lung metastasis was significantly reduced by co-injection of B16-BL6 cells with Ca-SP in a dose-dependent manner. Seven intermittent ⅰ.ⅴ. injection of 100$\mu\textrm{g}$ of Ca-SP caused a marked decrease of lung tumor colonization of B16-BL6 cells in a spontaneous lung metastasis model. These results suggest that Ca-SP, a novel sulfated polysaccharide, could reduce the lung colonization of B16-BL6 melanoma cells in experimental metastasis model, by inhibiting the tumor invasion of basement membrane Matrigel, probably through the prevention of the adhesion and migration of tumor cells to laminin-substrate and of the heparanase activity.

• BMB Reports
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• 제32권1호
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• pp.60-66
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• 1999

Matrix metalloproteinase-2 (MMP-2; 72-kDa gelatinase; 72-kDa type IV collagenase; gelatinase A) plays an important role in normal physiological processes and in many pathologic processes such as arthritis and metastasis of cancer. Tissue inhibitor of metalloproteinases-2 (TIMP-2) binds to proMMP-2 or mature MMP-2 at a 1:1 ratio and inhibits the catalytic activity of MMP-2. We demonstrated that the baculovirus/insect cell system does not have TIMP-2 activity. The human proMMP-2 free of TIMP-2 was expressed in the expression system and purified by one-step affinity chromatography using gelatin-Sepharose. The free proMMP-2 was autoactivated to the mature MMP-2 and autodegraded into smaller molecular weight forms in the absence of external activator. The activation and autodegradation of the proMMP-2 was much more rapid in the presence of 4-aminophenylmercuric acetate (APMA). Addition of TIMP-2 inhibits both APMA-induced activation and autodegradation of the free proMMP-2. However, an increasing concentration of TIMP-2 more readily inhibited activation of the free proMMP-2 than autodegradation. These results demonstrate that TIMP-2 plays roles in inhibition of both activation and autodegradation of the free proMMP-2 in addition to inhibition of the catalytic activity of MMP-2.

• 생명과학회지
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• 제15권2호
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• pp.231-235
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• 2005

기질 금속단백분해효소(MMP)는 기저막이나 간질성 조직 등에 있는 세포외기질 성분을 분해하여 상처 치유, 태아의 발생, 종양세포의 침윤과 전이 등을 포함하여 조직이 재구성되는 과정에서 생리학적 및 병리학적인 과정 양쪽 모두에 매우 중요한 역할을 한다. 본 연구에서는 MMP-9 유전자의 발현을 억제하는 물질을 천연물에서 탐색하였고, 탐색된 시료 중에서 높은 역가를 가진 낙우송과의 Metasequoia glyptostroboides가 선택되었다. 여러 가지 flavonoid 성분을 가지고 있는 Metasequoia glyptostroboides를 이용하여 4개의 biflavonoid와 2개의 monoflavonoid 구조의 물질을 분리하였고, 이 flavonoid들은 sciadopitysin, isoginkgetin, bilobetin, 2, 3-dihydrohino-kiflavone, luteolin, apigenin으로 정제하여 구조를 밝히고 zymography, WST-1을 이용한 세포독성시험, northern blot 등의 실험을 통하여 각 화합물의 구조적인 특성과 함께 MMP-9 유전자 발현을 억제하는 효과가 있는 것을 확인하였다.

• Preventive Nutrition and Food Science
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• 제13권3호
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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.

• The Korean Journal of Ecology
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• 제21권5_2호
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• pp.575-583
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• 1998

Physicochemical factors, microbial population size and the properties of the bacterial isolates were assessed to find out the nature of soil ecosystem of Mt. Paektu. Samples were obtained from the surface layer of soils on which specific plant community is developed. Average content of moisture, organic matter and avaiable phosphate of the soils were 21.6%, 17.3% and 2.48mg/100g, respectively. These values were similar to those of developing forest soils, but were slightly lower than those of climax ecosystem such as Piagol in Mt. Chiri. The population size of soil bacteria ranged from 2.7 to $202.5{\times}10^5$ CFU/g.dry soil, and the size is somewhat dependent on the content of moisture and oranic matter of the forest soil. A large number of bacteria was able to decompose macromolecules such as starch, elastin and gelatin. While the distribution rate of resistant bacteria to antibiotics was high, that to toxic chemicals was low. This means that the competition between microorgani는 predominate over the interference with artificial behaviour such as spread of pesticides in the surveyed region. Bacterial species composition of each soil was comparatively simple. Pseudomonas, Agrobacterium, Flavobacterium and Xanthomonas which are Gram-negative short rods were widely distributed in the forest soils. The endospore forming Bacillus species were also main constituents of the soil microflroa. any one of the strains was not identified as Azospirillum or Micrococcus which are known to be one of major constituents of the forest soil. for the correct identification of isolates chemotaxonomic studies will be proceeded, and the strains are to be stored in the Type collection Center.