• 생명과학회지
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• 제8권5호
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• pp.589-597
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• 1998

본 연구는 우리나라의 수산가공율의 증가에 따라 부수적으로 증가되고 있는 수산부산물 내지는 폐기물을 효율적으로 이용하려는 연구의 일환으로서 어류의 비늘에 대한 성분을 분석하고 아울러 해산어와 담수어 유래의 비늘 사이에 서식환경에 따른 함유성분의 차이가 있는지에 관하여 검토하였다. 해산어와 담수어의 두 어종간에 있어서 함유하고 있는 성분의 차이는 거의 없이 대부분 일치하였으나 함량의 차이는 다소 다르게 나타났다. 숭어비늘의 일반성분 함량은 회분과 단백질이 거의 50%로 같은 비율로 분포하고 있는데 반하여 잉어비늘에서는 회분이 약 20% 그리고 단백질이 약 80%로서 단백질의 함량이 회분보다 약 4배 가량 높았다. 비늘에서 추출한 단백질은 그 아미노산 조성 및 전기영동패턴에 의해 콜라겐으로 확인되었으며, 또한 비늘 콜라겐의 $\alpha$1 a 및 $\alpha$2 사슬의 전기이동패턴은 대조구로 사용된 소껍질의 Type I 콜라겐의 그것들과 거의 똑같은 것으로 보아 비늘 콜라겐도 Type I으로 추측되었다. 그리고 숭어비늘의 경우에는 함유하고 있는 대부분의 단백질이 콜라겐인데 비하여 잉어비늘은 전체 단백질 중 약 53%만을 차지하고 있었다. 비늘로부터 추출한 조회분에 대한 원소분석 및 X-ray 회절분석에서는 그 주성분 hydroxyapatite로 확인되었으며, 시료간의 차이는 전혀 없이 거의 완전하게 일치하였다.

• BMB Reports
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• 제35권6호
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• pp.576-582
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• 2002

Collagenase from the internal organs of a mackerel was purified using acetone precipitation, ion-exchange chromatography on a DEAE-Sephadex A-50, gel filtration chromatography on a Sephadex G-100, ion-exchange chromatography on DEAE-Sephacel, and gel filtration chromatography on a Sephadex G-75 column. The molecular mass of the purified enzyme was estimated to be 14.8 kDa by gel filtration and SDS-PAGE. The purification and yield were 39.5-fold and 0.1% when compared to those in the starting-crude extract. The optimum pH and temperature for the enzyme activity were around pH 7.5 and $55^{\circ}C$, respectively. The $K_m$ and $V_{max}$ of the enzyme for collagen Type I were approximately 1.1 mM and 2,343 U, respectively. The purified enzyme was strongly inhibited by $Hg^{2+}$, $Zn^{2+}$, PMSF, TLCK, and the soybean-trypsin inhibitor.

• Parasites, Hosts and Diseases
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• 제32권4호
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• pp.231-242
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• 1994

자연 감염된 가재에서 폐흡충의 피낭유충을 분리하고 개에 경구 감염시켜 성충을 얻었다. 폐흡충 성충의 조효소를 ion-exchange chromatography, affinity chromatography와 gel filtration chromatoglaphy를 실시하여 cysteine proteinase를 순수 정제하였다. 이들 효소의 생화학적 특성과 분해능을 관찰하였으며. 효소면역전기영동이적법을 이용하여 순수 정제한 효소의 항원성을 관찰하였다. 정제된 효소는 저분자 합성기질인 CBZ-arg-arg-AFC 보다 CBZ-phe-arg-AFC에서 높은 활성을 보였으며. 이들 효소는 thiol-dependent이었다. 정제된 효소 및 조효소의 최적 pH는 5.5이었고. 최적 mole 농도는 0.1 M(0.1 M sodium citrate, pH 5.5)이었고, 이들 효소는 $4^{\circ}C$에서 48시간 동안 80%의 안정성을 보였다 정제된 효소의 native 분자량은 20.000 dalton이었고, SDS- PAGE상에 나타난 분자량은 17,500 dalton이었다 정제된 효소는 cysteine proteinase 특이 억제 인자인 E-64, lodoacetic acid, NEM에 의해 활성이 완전히 억제되었으며, serine proteinase, aspartic proteinase 및 metallo proteinase 특이 억제인자에 의해 활성이 억제되지 않았다. 정제된 효소는 collagen(Type I)과 hemoglobin을 분해하였고, 효소면역전기영동이적법으로 정제된 효소의 항원성을 확인하였다.

• Preventive Nutrition and Food Science
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• 제15권2호
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• pp.143-146
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• 2010

Organic zinc was included in the diet of broiler chickens to examine its effect on the skin characteristics and the expression level of skin proteins. Broiler chicks (Ross$\times$Ross) were fed a corn-wheat-soybean meal basal diet, either as control or containing an additional 80 ppm of zinc proteinate for 4 weeks, and then five broilers from each treatment were selected randomly, slaughtered, and their skin characteristics were examined. There were significant increases (p<0.05) in thigh skin epidermis and dermis thickness in the chicks fed organic zinc. Collagen content in the skin of broilers was also increased by the addition of organic zinc to the diet. 2D-gel electrophoresis patterns indicated that expression levels of the three proteins, glyoxylase 1, hypothetical protein, and dispersin B were affected by zinc feeding. These results suggest that adding organic zinc to the chicken's feed may contribute to decreased skin tearing.

• Journal of Periodontal and Implant Science
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• 제41권5호
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• pp.227-233
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• 2011

Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.99-105
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• 2000

Cell-Cell interaction and the extracellular matrix (ECM) are belisved to play essential roles during in vitro culturing of primary hepatocytes in the control of differentiation and in the maintenance of tissue spcific functions. The objective of this study was to examine the effects of degree of cell-cell contact (DCC) on liver sperific function of rat promary hepatocytes. Hepatocyte aggregates with various with various degrees of cell-cell contantact, I. e., dispersed cell, longish aggregate, rugged aggregate, and smooth spheroid were obtained at 1, 5-6, 15-20, and 36-48 hrs, respectively in suspension cultures grown in spinner flasks embedded in Caalginate bead and collagen gel in order. The may result from mass transfer limitation and shear damage caused by agitation during aggregation. The rugged aggregate showed a higer viability and albumin secretion rate than the dispersed cells or the other aggregates. This result indicates the possible enhancement of a bioartificial liver's (BAL) performance using primary hepatocytes and the reduction in time to prepare a BAL through optimization of the immobilization time.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.590-600
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• 2003

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

• Journal of Microbiology and Biotechnology
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• 제25권3호
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• pp.399-406
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• 2015

In this study, we developed a composite filler comprising cross-linked hyaluronic acid (HA) and human collagen (COL) derived from the human umbilical cord with the aim of improving its biocompatibility and longevity compared with commercially available fillers. After HA/COL composite fillers were made in two different ratios (10:1 and 5:1), the physical properties of the fillers were evaluated. The interior morphologies and in vivo weight change of these hydrogels were also characterized at 1-16 weeks after injection into mice. To evaluate their biocompatibility and durability in vivo, we injected the composite fillers into nude mice subcutaneously. The variations of injected gel weight were measured and compared with the commercial dermal fillers (Restylane and TheraFill). The composites showed improved or similar physical properties (complex viscosity of 19-22 × 10⁵ cP, and injection force of 10-12 N) over the commercial dermal fillers. Sixteen weeks following the injection, the ratio of remaining composite filler weight to initial weight (75.5 ± 16.9%; 10:1) was shown to be greater than that of the commercial fillers (43.2 ± 8.1%, Restylane; 12.3 ± 5.3%, TheraFill). In addition, immunohistochemical analysis with angiogenesis-related markers such as isolectin and vWF revealed newly formed blood vessels and cellular influx into the composite filler, which were not observed in the other fillers. These results clearly suggest that the HA/COL composite filler is a superior candidate for soft tissue reconstruction. The filler we developed may be a suitable candidate as an injectable dermal filler for tissue augmentation in humans.

• 방사선산업학회지
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• 제9권4호
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• pp.199-207
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• 2015

Typical radiation cross-linked hydrogels has the characteristic that high water content, but low emission efficiency of active ingredients. Therefore, the hydrogel was prepared by the addition to collagen, which is closely related to the formation of skin wrinkles in biocompatibility and highly water-soluble carboxymethyl cellulose sodium salt (CMC) in order to preparation of hydrogels has excellent emission efficiency of active ingredients. Hydrogels were prepared by dissolving CMC and collagen each of 0.5%, 10% concentration in deionized water. Then, prepared hydrogels are performed by gamma-radiation at 1, 3, 5 kGy irradiation dose. The results showed that the gel fraction of after irradiated 3 kGy hydrogel was higher than before irradiated gelation as long as the 55.3%. The swelling rate of irradiated 3 kGy hydrogel was lower than the non-irradiated sample. The compressive strength of 3 kGy irradiated hydrogel was the highest. The visco-elastic did not show any significant differences, even after irradiation. The CMC hydrogel in this study suggested a potential use as a material for the mask pack for improved emission efficiency of the active ingredient and anti-wrinkles.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.799-821
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• 1996

The ultimate goal of periodontal treatment has been to facilitate regeneration of diseased periodontal tissues, destroyed by inflammatory periodontal disease. For regeneration of the periodontium to occur, all of component tissues must be restored to their original position and architecture. Growth factors which were known to promote the cellular processes, ie, proliferation, migration and matrix synthesis, have been in the spotlight of current periodontics. Platelet-derived growth factor(PDGF) stimulates collagen and non collagen protein synthesis, migration and proliferation of periodontal ligament cells. Insulin-like growth factor(IGF) has potentials to induce collagen and bone matrix synthesis so that it regulates normal bone remodeling. Application of the combination have been known to facilitate formation of bone and cementum, and to synergistically interact to promote coronal migration and proliferation of periodontal ligament cells. These two growth factors have been reported to exhibit positive effect in the periodontally diseased teeth or class m furcation defects. The aim of the present study was to test the hypothesis that PDGF-BB alone or the combination of PDGF-BB and IGF-I can predictably enhance regeneration of the periodontium in the dehiscence defect. Following the resection of premolars, roots were embedded. After 12 weeks of healing period, standardized experimental $4{\times}4mm$ dehiscence defects were created on the mid-facial of the premolar roots in each of 4 young adult dogs. In control group, only methylcellulose gel was inserted in the defects. In experimental group I and II, gel with $2{\mu}g$ of PDGF-BB or $2{\mu}g$ of PDGF-BB and $1{\mu}g$ of IGF-I was inserted in the defects, respectively. At 8 weeks postsurgery, the dogs were sacrificed. The results were observed histologically and analyzed histomorphometrically.The results of this study were as follws. 1. The new cementum formation was $1.26{\pm}0.69mm$ in the control group, $1.80{\pm}0.84mm$ in the experimental group I, $1.93{\pm}0.51mm$ in the experimental group II. The experimental group III, the experimental group I, the control group were in the order of cementum formation without statistically significant differences between control and all experimental groups. 2. The new bone formation was $1.00{\pm}0.53mm$ in the control group, $1.53{\pm}0.63mm$ in the experimental group I, $l.33{\pm}0.45mm$ in the experimental group II. The experimental group I, the experimental group II, the control group were in the order of bone formation without statistically significant differences between control and all experimental groups. 3. The root resorption was $1.12{\pm}0.64mm$ in the control group, $1.34{\pm}0.73mm$ in the experimental group I, $0.79{\pm}0.59mm$ in the experimental group II without statistically significant differences between control and all experimental groups. These results suggested that the use of PDGF-BB alone or PDGF-BB and IGF-I in the dehiscence defects might facilitate periodontal regeneration in some degree, but has not shown statistically significant results.