• Journal of Animal Science and Technology
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• v.45 no.3
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• pp.491-498
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• 2003

The purpose of this study is to observe purification and properties of osteopontin(OPN) from bovine milk. The purification of osteopontin from bovine milk was performed by using ion-exchange and hydrophobic chromatography. SDS-PAGE analysis revealed that the protein migrated at Mw. 60,000. NH2-terminal sequence analysis of the first seven amio acids revealed the protein to be identical to that previously reported for bovine OPN. 35-wk-old chickens, including 3 Single Comb White Leghorn (SCWL), were used to produce egg yolk antibody(IgY) against OPNas a antigen. However, the anti-OPN antibody activities determined by ELISA. Immunological assy of OPN in milk was performed using radial immunodiffusion test based on the standard curve of pure OPN. The radial precipitation lines of four different milk samples indicated that the concentrations of OPN in the milk samples were within the range of 31.7 to 39.7${\mu}g$/ml. On inhibition with OPN on precipitation of calcium phosphate, OPN was slightly higher than casein phosphopeptide(CPP) and poly-glutamic acid.

• Journal of Plant Biotechnology
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• v.37 no.2
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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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• v.3 no.4
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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.

• The korean journal of orthodontics
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• v.44 no.3
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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.

• Food Science of Animal Resources
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• v.41 no.4
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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.

• Journal of Korean Dental Hygiene Science
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• v.5 no.1
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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.

• Journal of the korean academy of Pediatric Dentistry
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• v.37 no.1
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• pp.1-12
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• 2010

The aim of this study was to evaluate the preventive effect of commercially available anticariogenic products, specifically, the tooth cream containing Casein phosphopeptide-amorphous calcium phosphate(CPP-ACP), fluoride varnish and low-level fluoride mouthrinse on enamel erosion induced by carbonated beverage in a short period of time. Enamel specimens were treated as follows and were then kept in artificial saliva for 24 hours followed by further processing by alternately soaking them in Cola beverage and in distilled water for 1 minute each five times. Group 1: control group (no treatment) Group 2: tooth cream with CPP-ACP Group 3: fluoride varnish (1,000 ppm F) Group 4: low-level fluoride mouthrinse (227 ppm F) Group 5: fluoride varnish + tooth cream with CPP-ACP Group 6: low-level fluoride mouthrinse + tooth cream with CPP-ACP Microhardness and erosion depth were measured and the mineral loss of each specimen was evaluated by measuring the volumetric fluorescence change(${\Delta}Q$) against the stable fluorescent grid using quantitative light-induced fluorescence(QLF). The experiment lasted for 6 days repeated each day. The results were as follows: 1. The microhardness was increased as follows: Group $1{\leq}2{\leq}4$<6<$3{\fallingdotseq}5$. 2. The mean erosion depth was increased as follows: Group $5{\fallingdotseq}3$<6<$4{\fallingdotseq}2{\fallingdotseq}1$. 3. The ${\Delta}Q$ was increased as follows: Group $1{\fallingdotseq}2{\leq}4{\leq}6{\leq}3{\fallingdotseq}5$. The decrement of ${\Delta}Q$ was similar between group 1 and 2, group 4 and 6 and group 3 and 5. 4. The ${\Delta}Q$ showed positive correlation with microhardness (r=0.96, p<0.05), while it was negatively correlated to erosion depth (r=-0.96, p<0.05).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.713-717
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• 1999

In order to utilize protein hydrolysate produced from hoki (Johnius Belengeri) frame among many different fish processing wastes, hoki frame peptide (PHFP) and phosphorylated hoki frame peptide (PHFP) were prepared, and their calcium absorption accelerating effects were investigated in comparison to control and casein phosphopeptide (CPP). In in vitro experiment, HFP and PHFP inhibited calcium phosphate formation as high as 1.5-fold and 2.5-fold, respectively, comparing to control, In addition, the inhibition rate of calcium phosphate precipitation as increasing concentrations of HFP and PHFP was risen and was similar to that of CPP at 2.0 mg/ml of PHFP concentration, In in vivo experiment using the rats, the groups fed HFP and PHFP indicated significantly increased calcium content in the femur. In particular, the calcium content in the small intestine of the rat fed PHFP was higher than that of control group by approximately $60\%$.

• Journal of the korean academy of Pediatric Dentistry
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• v.39 no.2
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• pp.129-138
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• 2012

The aim of this in vitro study was to evaluate the effectiveness of experimental 2.26% fluoride-polyvinyl alcohol (F-PVA) tape in inhibition of enamel demineralization using enamel surface microhardness (SMH) analysis and scanning electron microscopy (SEM) examination. Enamel specimens (n=60) randomly assigned to four groups: control group, F-PVA tape group, fluoride varinish (F-varnish) group, casein phosphopeptide-amorphous calcium phosphate (CPP-ACFP) group. After topical application, pH-cycling was processed. Then, SMH was measured and the percentage loss of surface microhardness (%SML) was calculated. For the SEM examination, five sample specimens in each group were treated and the morphologic character was evaluated. After pH-cycling, the SMH values of the enamel specimens of F-PVA tape and F-varnish group were significantly higher than that of CPP-ACFP group, there was no significant difference between F-PVA tape and Fvarnish group. With SEM examination, enamel surfaces in the F-PVA tape group and F-varnish group showed numerous spherical and ovoid crystals formed on the enamel surface were also observed. The density of crystals was higher than that of both control group and CPP-ACFP group. F-PVA tape is effective in inhibition of enamel demineralization. Also, F-PVA tape's inhibition of enamel demineralization is comparable to that of F-vanish and greater than that of CPP-ACFP.