• 한국토양동물학회지
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• 제4권2호
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• pp.101-106
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• 1999

지렁이 꼬리재생 초기에 발현되는 피브리노겐 분해효소의 활성은 세포외기질의 재구성에 연관되어 있을 것으로 생각된다. 본 연구에서는 지렁이의 꼬리 재생 중에 발현된 피브리노겐 분해효소의 활성과 특성을 밝히고자 하였다. 지렁이의 꼬리 재생 중에 발현되는 피브리노겐 분해효소는 최소 7개이며 그 분자량은 각각 58, 45, 32, 27, 23, 19 그리고 12kDa로 측정되었다. 피브리노겐 분해효소의 활성은 절단 후 1일부터 효소의 활성이 나타나서 7일까지 거의 유사한 정도의 활성이 유지되었으며, 7일 이후부터는 이 효소들의 활성이 급격히 감소하여 14일에는 그 활성이 대조군 수준으로 회복되었다. 지렁이 꼬리 재생 중 발현된 모든 피브리노겐 분해효소는 PMSF와 aprotinin를 처리하였을 때 효소들의 활성이 억제되었다. 또한 Ca$^{2+}$의 제거는 이효소의 활성에 영향을 미치지 않는 것으로 나타났다. 이러한 결과를 통하여 지렁이 꼬리재생시 발현되는 피브리노겐 분해효소는 serine계 단백질 분해효소임을 알 수 있었다.

• Molecules and Cells
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• 제28권3호
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• pp.183-188
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• 2009

TSAd/Lad is a T cell adaptor molecule involved in $p56^{lck}$-mediated T cell activation. To investigate the functions of TSAd in T cells, we generated transgenic (TG) mice expressing the SH2 domain of TSAd (TSAd-SH2) under the control of the $p56^{lck}$ proximal promoter. In T cells from TSAd-SH2 TG mice, T cell receptor (TCR)-mediated early signaling events, such as $Ca^{2+}$ flux and ERK activation, were normal; however, late activation events, such as IL-2 production and proliferation, were significantly reduced. Moreover, TCR-induced cell adhesion to extracellular matrix (ECM) proteins and migration through ECM proteins were defective in T cells from TSAd-SH2 TG mice. Furthermore, the contact hypersensitivity (CHS) reaction, an inflammatory response mainly mediated by T helper 1 (Th1) cells, was inhibited in TSAd-SH2 TG mice. Taken together, these results show that TSAd, particularly the SH2 domain of TSAd, is essential for the effector functions of T cells.

• BMB Reports
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• 제49권2호
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• pp.116-121
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• 2016

Although proteomic analyses have revealed the presence of mitochondrial oxidative phosphorylation (OXPHOS) proteins in the plasma membrane, there have been no in-depth evaluations of the presence or function of OXPHOS I-V in the plasma membrane. Here, we demonstrate the in situ localization of OXPHOS I-V complexes to the sarcolemma of skeletal muscle by immunofluorescence and immunohistochemistry. A portion of the OXPHOS I-V complex proteins was not co-stained with MitoTracker but co-localized with caveolin-3 in the sarcolemma of mouse gastrocnemius. Mitochondrial matrix-facing OXPHOS complex subunits were ectopically expressed in the sarcolemma of the non-permeabilized muscle fibers and C2C12 myotubes. The sarcolemmal localization of cytochrome c was also observed from mouse gastrocnemius muscles and C2C12 myotubes, as determined by confocal and total internal resonance fluorescence (TIRF) microscopy. Based on these data, we conclude that a portion of OXPHOS complexes is localized in the sarcolemma of skeletal muscle and may have non-canonical functions.

• Parasites, Hosts and Diseases
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• 제41권4호
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• pp.189-196
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• 2003

In order to evaluate the possible roles of secretory proteases in the pathogenesis of amoebic keratitis, we purified and characterized a serine protease secreted by Acanthamoeba lugdunensis KA/E2, isolated from a Korean keratitis patient The ammonium sulfate-precipitated culture supernatant of the isolate was purified by sequential chromatography on CM-Sepharose, Sephacryl S-200, and mono Q-anion exchange column. The purified 33 kDa protease had a pH optimum of 8.5 and a temperature optimum of $55^{\circ}C$. Phenylmethylsulfonylfluoride and 4-(2-Aminoethyl)-benzenesulfonyl-fluoride, both serine protease specific inhibitors, inhibited almost completely the activity of the 33 kDa protease whereas other classes of inhibitors did not affect its activity. The 33 kDa enzyme degraded various extracellular matrix proteins and serum proteins. Our results strongly suggest that the 33 kDa serine protease secreted from this keratopathogenic Acanthamoeba play important roles in the pathogenesis of amoebic keratitis, such as in corneal tissue invasion, immune evasion and nutrient uptake.

• BMB Reports
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• 제30권2호
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• pp.106-112
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• 1997

The effect of cryopreservation on extracellular matrix was studied with the ultimate objective of permiting a prediction of the tendency of aorta conduit tissue to calcify following transplantation. Cryopreserved and fresh porcine aorta conduit tissues were extracted using guanidine-hydrochloride (Gdn-HCl) followed by sequential digestion of the tissues with collagenase, elastase, and papain. Glycosaminoglycans (GAGs) of the proteoglycans (PGs) were isolated and quantitated. Gdn-HCl extracted about 61% and 62% of the total GAG (proteoqlycan) material from cryopreserved and fresh tissues, respectively. Collagenasesolubilized proteoglycans from Gdn-HCl extracted tissue represented 20% and 13%, respectively, of the total GAGs present in cryopreserved and fresh tissues. Subsequent elastase hydrolysis of collagenase-digested tissue released about 11% of total GAGs from cryopreserved tissue and 16% from fresh tissue. The remaining 8%, from cryopreserved tissue, and 9%, from fresh tissue, of the total GAGs were obtained after using a papain hydrolysis. There was essentially no difference between fresh and cryopreserved tissues in the relative distribution of proteoglycans in the extracts and digestions except in the initial digestion step where more proteoglycans were obtained from collagenase solubilization of cryopreserved tissue than fresh tissue (p<0.05). The histologic status of the fresh and cryopreserved porcine aortic conduit did not differ markedly. The normal tissue architecture was not affected markedly by the cryopreservation procedure as neither alteration of elastic structure, fibrous proteins nor alteration of nuclear distribution or smooth muscle cell morphology was detected. Quantitative tissue mineral studies revealed that the mean calcium content of the cryopreserved aorta conduit tissue $(165{\pm}3\;{\mu}g/g\;wet\;tissue)$ was higher than that of the fresh tissue $(105{\pm}4\;{\mu}g/g\;wet\;tissue)$ $(p<0.05)$. The mean phosphorus content was $703{\pm}35\;{\mu}g$ wet tissue from cryopreserved tissue and $720{\pm}26\;{\mu}g$ wet tissue from fresh tissue. The study indicates that there is no significant alteration in the distribution of PGs in properly cryopreserved tissue, but the total calcium level appears to be increased in tissue cryopreserved by the cryopreservation process used in this study.

• 대한화장품학회지
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• 제30권4호
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• pp.463-470
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• 2004

인공피부배양물(DA-3711)은 동아제약의 인공피부배양기술을 활용하여 개발되었으며, 노화된 피부상태를 개선시켜 줄 수 있는 천연의 세포외기질 단백질을 비롯하여 인체성장인자 등의 영양성분을 함유하고 있다 DA-3711의 항노화 효과는 in vitro와 in vivo에서 규명되었으며, 또한 인체 효능 연구 결과에서도 피부탄력을 개선시키고 주름을 감소시키는데 매우 효과적이었다. 즉, DA-3711을 이용한 새로운 주름개선제는 피부재생을 촉진시켜 항노화 및 주름개선 효능을 나타낸다.

• Molecules and Cells
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• 제41권12호
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• pp.1016-1023
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• 2018

Regenerative orthopedics needs significant devices to transplant human stem cells into damaged tissue and encourage automatic growth into replacements suitable for the human skeleton. Soft biomaterials have similarities in mechanical, structural and architectural properties to natural extracellular matrix (ECM), but often lack essential ECM molecules and signals. Here we engineer mineralized polysaccharide beads to transform MSCs into osteogenic cells and osteoid tissue for transplantation. Bone morphogenic proteins (BMP-2) and indispensable ECM proteins both directed differentiation inside alginate beads. Laminin and collagen IV basement membrane matrix proteins fixed and organized MSCs onto the alginate matrix, and BMP-2 drove differentiation, osteoid tissue self-assembly, and small-scale mineralization. Augmentation of alginate is necessary, and we showed that a few rationally selected small proteins from the basement membrane (BM) compartment of the ECM were sufficient to up-regulate cell expression of Runx-2 and osteocalcin for osteoid formation, resulting in Alizarin red-positive mineral nodules. More significantly, nested BMP-2 and BM beads added to a non-union skull defect, self-generated osteoid expressing osteopontin (OPN) and osteocalcin (OCN) in a chain along the defect, at only four weeks, establishing a framework for complete regeneration expected in 6 and 12 weeks. Alginate beads are beneficial surgical devices for transplanting therapeutic cells in programmed (by the ECM components and alginate-chitosan properties) reaction environments ideal for promoting bone tissue.

• 생명과학회지
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• 제20권4호
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• pp.602-606
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• 2010

제2형 당뇨병은 복합적인 유전적 경향을 보이며, 당뇨병과 관련이 있는 것으로 알려진 여러 대립유전자들이 보고되어 있다. Matrix metalloproteinase (MMP)는 기질 금속단백효소로 세포외기질(extracellular matrix, ECM)을 선택적으로 분해하는 효소로서, 질환의 진행뿐만 아니라, 배 발생, 조직형성, 염증반응, 상처치유 등을 조절하는 것으로 알려져 있다. 이에 본 연구에서는 한국인에서 MMP3의 A(-267)G, A658G, T813C 유전자다형성이 제2형 당뇨병 환자와 어떤 연관성을 갖는지 알아보고자 하였다. 정상대조군 100명(남자 36명, 여자 64명)과 당뇨병 환자 200명(남자 108명, 여자 92명)을 대상으로 하였다. 그 결과 A(-267)G와 A658G 유전자다형성이 제2형 당뇨병과 연관이 있는 것으로 나타났기에, 제2형 당뇨병의 발병과 관련이 있을 것으로 생각된다. 향후 대상 환자 수와 대조군을 더욱 늘리는 등의 추가적인 연구를 할 필요가 있을 것으로 생각되며, DNA chip과 같은 유전자 수준에서의 진단법으로 발전시킬 수 있을 것이다.

• Tuberculosis and Respiratory Diseases
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• 제64권1호
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• pp.22-27
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• 2008

연구배경: 본 연구에서는 황색포도알균이 숙주세포 내로 침입하여 증식하는 정도를 관찰하고자 한다. 이때 세포 외 바탕 단백질의 변화가 수반될 것으로 가설을 설정하고, 이러한 변화에 영향을 미칠 것으로 생각되는 matrix metalloproteinase (MMP)의 발현과 역할에 대해 연구하고자 하였다. 방법: 황색포도알균은 $10^6{\sim}10^7CFU/ml$을 $10^5$개의 기관지상피세포인 BEAS-2B 세포에 2시간 동안 침입시킨다. 이후 세척으로 세포 밖에 있는 황색포도알균을 제거한 후, BEAS-2B 세포를 다양한 시간 동안(4, 6, 8, 12 시간) 배양한 후 황색포도알균의 집락수(CFU/ml)를 측정하였고, 단백질을 분리하여 세포 외 바탕단백질의 발현 정도와 MMP의 활성도를 측정하였다. 또한 MMP 억제제인 GM6001을 전처치한 후 황색포도알균을 세포에 침입시킨 후 세포 내에서의 균의 집락수 및 세포 외 바탕단백질의 변화를 관찰하였다. 결과: 황색포도알균의 집락을 측정한 결과 4시간과 12시간을 비교해 볼 때 MOI가 증가할수록, 감염시킨 시간이 길수록 숙주세포 내로 침입이 유의하게 증가하였다. BEAS-2B 세포에서 황색포도알균을 침입시킨 시간이 길수록, MOI가 증가할수록 MMP 2 및 MMP 9의 활성도와 dysadherin의 발현은 증가하였고, 이와는 대조적으로 E-cadherin의 발현은 감소하였다. MMP억제제인 GM6001을 전 처치 한 결과 황색포도알균의 세포 내 침입을 유의하게 감소시켰다. 결론: 황색포도알균이 기관지 상피세포 내로 침입할 때 dysadherin 및 E-cadherin 같은 세포 외 바탕 단백질의 변화를 동반하며, MMP 활성도가 균의 세포 내 침입에 관여하는 것으로 보인다.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2011년도 추계학술발표대회
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• pp.2.2-2.2
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• 2011

The central strategy in tissue engineering involves a biomaterial scaffold as a delivery carrier of cells and a depot to deliver bioactive molecules. The ability of scaffolds to control cellular response to direct particular repair and regeneration processes is essential to obtain functional tissue engineering constructs. Therefore, many efforts have been made to understand local interactions of cells with their extracellular matrix (ECM) microenvironment and exploit these interactions for designing an ideal scaffold mimicking the chemical, physiological, and structural features of native ECM. ECM is composed of a number of biomacromolecules including proteins, glycosaminoglycans, and proteoglycans, which are assembled together to form complex 3-dimensional network. Electrospinning is a process to generate highly porous 3-dimensional fibrous structure with nano to micro scaled-diameter, which can closely mimic the structure of ECM. In this presentation, our approaches to develop biomimetic electrospun fibers for modulation of cell function will be discussed.