• Journal of Animal Science and Technology
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• 제45권3호
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• pp.491-498
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• 2003

본 연구는 우유로부터 OPN을 분리 정제하여 그 특성을 규명하기 위해서 수행되었다. 먼저 ion-exchange와 hydrophobic chromatography를 이용하여 우유로부터 OPN을 분리 정제하였다. OPN의 분자량은 SDS-전기영동 상에서 약 60,000dalton이었고, NH2-terminal amino acid sequence를 확인한 결과 Leu-Pro-Val-Lys-Pro- Thr-Ser 순이었다. OPN을 35주령된 SCWL를 이용하여 산란계를 면역 시키고, 형성된 anti- OPN IgY 항체를 분리․정제한 후 ELISA test로 항체가를 측정하였다. 또한 RID test을 이용하여 OPN 함량에 따른 정량곡선을 작성하고, 이 곡선에 의해 우유 중 OPN 함량을 정량하였다. 그 결과 원유, 탈지유, 시유에서 각각39.78, 31.74, 37.48${\mu}g$/$m\ell$을 함유하고 있는 것으로 나타났다. 또한 OPN의 Ca 가용화 능력을 검정한 결과 OPN이 CPP와 poly-glutamic acid 보다 더 우수한 것으로 나타났다.

• Journal of Plant Biotechnology
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• 제37권2호
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• pp.196-204
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• 2010

With the completion of genome sequencing of several organisms, attention has been focused to determine the function and functional network of proteins by proteome analysis. The recent techniques of proteomics have been advanced quickly so that the high-throughput and systematic analyses of cellular proteins are enabled in combination with bioinformatics tools. Furthermore, the development of proteomic techniques helps to elucidate the functions of proteins under stress or diseased condition, resulting in the discovery of biomarkers responsible for the biological stimuli. Ultimate goal of proteomics orients toward the entire proteome of life, subcellular localization, biochemical activities, and their regulation. Comprehensive analysis strategies of proteomics can be classified as three categories: (i) protein separation by 2-dimensional gel electrophoresis (2-DE) or liquid chromatography (LC), (ii) protein identification by either Edman sequencing or mass spectrometry (MS), and (iii) quanitation of proteome. Currently MS-based proteomics turns shiftly from qualitative proteome analysis by 2-DE or 2D-LC coupled with off-line matrix assisted laser desorption ionization (MALDI) and on-line electrospray ionization (ESI) MS, respectively, to quantitative proteome analysis. Some new techniques which include top-down mass spectrometry and tandem affinity purification have emerged. The in vitro quantitative proteomic techniques include differential gel electrophoresis with fluorescence dyes, protein-labeling tagging with isotope-coded affinity tag, and peptide-labeling tagging with isobaric tags for relative and absolute quantitation. In addition, stable isotope labeled amino acid can be in vivo labeled into live culture cells through metabolic incorporation. MS-based proteomics extends to detect the phosphopeptide mapping of biologically crucial protein known as one of post-translational modification. These complementary proteomic techniques contribute to not only the understanding of basic biological function but also the application to the applied sciences for industry.

• Mass Spectrometry Letters
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• 제3권4호
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• pp.93-100
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• 2012

Cellular processes such as proliferation, differentiation, and adaptation to environmental changes are regulated by protein phosphorylation. In order to enhance the understanding of molecular dynamics for biological process in detail, it is necessary to develop sensitive and comprehensive analytical methods for the determination of protein phosphorylation. Neural stem cells hold great promise for neural repair following an injury or disease. In this study, we made differentiated oligodendrocytes from human neural stem cells using over-expression of olig2 gene. We confirmed using quantitative phosphoproteome analysis approach that combines stable isotope labeling by amino acids in cell culture (SILAC) and $TiO_2$ micro-column for phosphopeptide enrichment with $MS^2$ and $MS^3$ mass spectrometry. We detected 275 phosphopeptides which were modulated at least 2-fold between human neural stem cells and oligodendrocytes. Among them, 23 phosphoproteins were up-regulated in oligodendrocytes and 79 phosphoproteins were up-regulated in F3 cells.

• 대한치과교정학회지
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• 제44권3호
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• pp.113-118
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• 2014

Objective: The purpose of this in vitro study was to examine the effects of fluoridated, casein phosphopeptide-amorphous calcium phosphate complex (CPP-ACP)-containing, and functionalized ${\beta}$-tricalcium phosphate (fTCP)-containing toothpastes on remineralization of white spot lesions (WSLs) by using Quantitative light-induced fluorescence (QLF-D) Biluminator$^{TM}$ 2. Methods: Forty-eight premolars, extracted for orthodontic reasons from 12 patients, with artificially induced WSLs were randomly and equally assigned to four treatment groups: fluoride (1,000 ppm), CPP-ACP, fTCP (with sodium fluoride), and control (deionized water) groups. Specimens were treated twice daily for 2 weeks and stored in saliva solution (1:1 mixture of artificial and human stimulated saliva) otherwise. QLF-D Biluminator$^{TM}$ 2 was used to measure changes in fluorescence, indicating alterations in the mineral contents of the WSLs, immediately before and after the 2 weeks of treatment. Results: Fluorescence greatly increased in the fTCP and CPP-ACP groups compared with the fluoride and control groups, which did not show significant differences. Conclusions: fTCP- and CPP-ACP-containing toothpastes seem to be more effective in reducing WSLs than 1,000-ppm fluoride-containing toothpastes.

• 한국축산식품학회지
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• 제41권4호
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• pp.687-700
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• 2021

This study performed to evaluate the applicability of functional dairy food materials by comparing the calcium solubilization ability and anti-inflammatory effects of hydrolyzed casein protein. Commercial enzyme (Alcalase^®; Neutrase^®; Protamex^®; Flavourzyme^®) was added to the 10% casein solution to prepare the casein hydrolysates. Samples obtained every hour [1:200 (w/v)]. According to results of measuring the degree of hydrolysis (DH), all of four enzymatic hydrolysates increased rapidly from 30 to 40 min, and after 150 min, there were no change. Protamex^® and Neutrase^® had the highest DH compared to others enzymatic hydrolysates. After that, peptides obtained throughout a preparative liquid chromatography system. In the calcium solubility experiments, neutrase fraction (NF) 4 and NF7 showed similar activities with casein phosphopeptide (CPP). In vitro cell experiments showed that no cytotoxicity except for NF6. Also, the production of nitric oxide (NO) inhibited as the concentration of fraction samples increased. The cytokine (IL-1α, IL-6, and TNF-α) production was lower than lipopolysaccharide (+) group significantly. Therefore, the possibility of anti-inflammatory activity found in the hydrolyzed samples. According to the above experiments, NF3 and Protamex Fraction (PF) 3 selected. Amino acids selected throughout an AccQ-Tag system. As a result, 17 species of amino acids and several species of unknown amino acids identified. Both fractions had the highest content of phenylalanine. This study identified the potential of biologically active and functional peptides derived from casein that affect the food and dairy industry.

• 대한치위생과학회지
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• 제5권1호
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• pp.29-38
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• 2022

Background: This study aimed to compare the remineralization effects of sodium caseinate and other substances on artificially demineralized enamel. Methods: We selected 25 healthy human premolars and molars and produced a total of 75 specimens by dividing them into five groups: control group, with distilled water; experimental group 1 (EG1), with 3% sodium caseinate; EG2, with 10% sodium caseinate; EG3, with casein phosphopeptide-amorphous calcium phosphate (CPP-ACP); and EG4, with 0.05% NaF. Subsequently, the specimens were immersed in an artificial demineralization solution for 60 min. The demineralized specimens were then immersed in a remineralization solution for 7 days. Surface microhardness was measured using a microhardness tester, and remineralized lesions were observed under a scanning electron microscope (SEM). Regarding statistical analysis, the paired t-test and analysis of variance were performed using the SPSS program. Results: Although the surface hardness of the remineralized lesions increased significantly in all groups (p<0.05), the average increment did not differ significantly between the groups. The surface microhardness of CPP-ACP was the highest, followed by that of 0.05% NaF and 10% sodium caseinate. The remineralization effect of sodium caseinate was similar to that of 0.05% NaF. SEM confirmed that all groups treated with the remineralization solution were remineralized. Conclusions: Although the remineralization effect of sodium caseinate was slightly lower than that of CPP-ACP, it was similar to that of 0.05% NaF. Therefore, to enhance the remineralization effect of sodium caseinate, the appropriate concentration and application time should be determined.

• 대한소아치과학회지
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• 제37권1호
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• pp.1-12
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• 2010

본 연구는 항우식 제품인 Casein phosphopeptide-amorphous calcium phosphate(CPP-ACP)를 포함하는 치아크림, 불소 바니쉬, 저농도 불소 양치액을 치아에 도포하는 것이 단시간 탄산음료에 의한 법랑질의 침식을 예방할 수 있는지 살펴보기 위해 계획되었다. 법랑질 시편에 다음과 같은 항우식 제품을 도포하였고, 인공타액에 24시간 보관한 후, 콜라에 1분 증류수에 1분씩 5회 번갈아 처리하였다. 1군: 대조군(무처치) 2군: CPP-ACP 치아크림군 3군: 불소 바니쉬군(1,000 ppm F) 4군: 저농도 불소 양치액군(227 ppm F) 5군: 불소 바니쉬 + CPP-ACP 치아크림군 6군: 저농도 불소 양치액 + CPP-ACP 치아크림군 미세경도와 침식깊이를 측정하였고, Quantitative light-induced fluorescence(QLF)를 이용하여 안정된 형광 격자에 대한 부피 형광 변화인 ${\Delta}Q$를 측정하여 무기질 감소량을 평가하였다. 6일 동안 실험을 반복하여 다음과 같은 결과를 얻었다. 1. 미세경도는 1군$\leq$2군$\leq$4군<6군<3군$\fallingdotseq$5군 순이었다. 2. 평균 침식깊이는 5군$\fallingdotseq$3군<6군<4군$\fallingdotseq$2군$\fallingdotseq$1군 순이었다. 3. ${\Delta}Q$는 1군$\fallingdotseq$2군$\leq$4군$\leq$6군$\leq$3군$\fallingdotseq$5군 순이었다. ${\Delta}Q$의 감소율은 1군과 2군, 4군과 6군, 3군과 5군이 각각 유사하였다. 4. ${\Delta}Q$는 미세경도와 강한 양의 상관관계를 나타냈고(r=0.96, p<0.05), 침식깊이와는 강한 음의 상관관계를 보였다(r=-0.96, p<0.05).

• 한국수산과학회지
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• 제32권6호
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• pp.713-717
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• 1999

수산가공공장에서 부산물로 대량 생산되고 있는 민태 frame 단백질로부터 회수된 펩타이드의 이용성을 탐색할 목적으로 민태 frame 단백질 유래 펩타이드와 이를 인산화시킨 펩타이드에 대한칼슘흡수 촉진효과를 칼슘의 침전형성 저지율을 이용한 in vitro 실험과 칼슘 결핍 흰쥐를 이용한 in vivo실험으로 검토하였다. 먼저 in vitro 실험에서 칼슘의 침전형성 반응시간의 변화에 따른 상충액중의 칼슘농도는 민태 frame 펩타이드(HFP)와 인산화시킨 펩타이드 (PHFP)를 첨가한 경우 대조군보다 각각 약 1.5배 및 2.5배 높은 것으로 나타났으며, 펩타이드의 첨가농도에 따른 실험에서도 HFP는 농도가 1.0mg/ml일 때 가용화율이 $70\%$로 증가하였고, PHFP는 0.8mg/ml일 때 약 $75\%$, 그리고 2.0mg/ml일 때는 CPP와 같은 $100\%$로 증가하였다. 흰쥐를 이용한 in vivo 실험에서 HFP 및 PHFP 섭취군 모두에서 대퇴골내 칼슘농도가 유의적으로 상승하였으며, 특히 PHFP 섭취군은 소장내 가용성 칼슘의 함유량이 대조군에 비해 약 $60\%$ 이상 높았다. 또 소변으로 배출되는 칼슘의 양은 HFP를 섭취한 군이 대조군보다 약 1.5배 높았다. 이들의 결과로부터 어류가공 부산물로부터 단백질 유래의 칼슘흡수 촉진제의 개발이 가능할 것으로 기대된다.

• 대한소아치과학회지
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• 제39권2호
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• pp.129-138
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• 2012

본 연구의 목적은 법랑질 표면 미세경도의 측정과 주사전자현미경 관찰을 통해 본 교실에서 개발한 2.26%의 불소를 함유한 폴리비닐알코올 테잎 (F-PVA tape)의 법랑질 탈회 억제 효과를 평가하기 위한 것이다. 60개의 법랑질 표본을 각 군당 15개 씩, 임의로 대조군, F-PVA tape 군, 불소바니쉬 군, CPP-ACFP 군으로 나누고, 각각의 재광화 제재를 도포한 후 pH-cycling 과정 거쳤다. 표본의 표면 미세경도를 측정하고 표면 미세경도 소실량을 계산하였다. 주사전자현미경 관찰을 위해, 각 군에서 5개의 표본을 추출하고 형태학적 특징을 분석하였고 다음과 같은 결론을 얻었다. 1. pH-cycling 후, F-PVA tape 군과 불소바니쉬 군의 법랑질 표면 미세경도 값은 CPP-ACFP 군과 비교 시 유의하게 높았으며, F-PVA tape 군과 불소바니쉬 군 사이에는 유의한 차이가 없었다. 2. 주사전자현미경 관찰 시, F-PVA tape 군과 불소바니쉬 군에서는 법랑질 표면에 형성된 수많은 구형 및 난원형의 결정 입자들이 관찰되었으며, 결정 입자의 밀도는 대조군 및 CPP-ACFP 군에서 보다 높았다. F-PVA tape은 법랑질의 탈회를 억제하는데 효과적이며, 그 효과는 불소바니쉬와 견줄만하고, CPP-ACFP 보다 우수하다.