• The Journal of the Korean dental association
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• v.12 no.7
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• pp.497-504
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• 1974

• 대한치과교정학회:학술대회논문집
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• 1998.10a
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• pp.79.3-79.3
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• 1998

• The Journal of the Korean dental association
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• v.20 no.9 s.160
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• pp.781-788
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• 1982

• Korean Journal of Oriental Medicine
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• v.1 no.1
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• pp.401-418
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• 1995

Aging process can be explained by many factors. In this study, we counted complete Blood Cells (CBC) such as WBC, Lymphocytes, Monocytes, Granulocytes, RBC, HGB, and HCT of both SAM P6 and SAM P1 during the aging process. Plasma albumin, glucose, alkaline phosphatase, calcium, creatinine, inorganic phosphate, urea concentrations were also measured at the same time. In addition to these, plasma concentrations of cortisol, total T3, and total T4 were analyzed by Chemiluminescent Immunoassay. There were no change in CBC counts of SAM R1 and SAM P6 during this study. Plasma concentratins of albumin and glucose decreased significantly in SAM R1. However, plasma alkaline phosphatases and creatinine concentration in SAM P6 decreased significantly at 16 week after birth comparing to the control. Total T4 levels were siginificantly increased although cortisol and total T3 concentrations were the same in SAM R1 groups. Especially, the after birth of creatinine, alkaline phosphatase, T4 of SAM P6 at 16 week were significantly different form those of SAM R1. At 12 week after birth, pilose antler extract was given 5g/kg/day p.o. for 0, 7, 14, 21, and 30 days each in both SAM R1 and SAM P6. The RBC, HGB, and HCT levels started to increase significantly from 7 days after the dose at SAM P6 only. Total T4 conectrations were elevated gradually during the study although the antler extract administration did not prevent or inhibit the increase in total T4 concentration during the study. Therefore, the elevation of erythrocytes after administration of the extract needs to be studied in future.

• Korean Journal of Animal Reproduction
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• v.19 no.1
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• pp.55-63
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• 1995

The putative totipotency germ cells has a relative abundance of alkaline phosphatases. Thus, histological staining of AP activity offers a new route to isolate totipotent cells and also provides insights into culture systems of these cells. Furthermore, the AP staining technique is simple and fast, requires only the napthol AS/MS substrate in combination with trapping diazonium salts such as fast red or fast blue. However, our unexpected finding was that AP staining of mouse ES cells were detected in the undifferentiaed epiblast-derived cells as well as several types of differentiating cells. This findings are different from results of Talbot et al. (1993) reported usefulness of the AP staining and implies that histological staining of AP may not by useful to determine undifferentiaed state or totipotency of ES cells. Thus, we have investigated the patterns of AP expression by RT-PCR in order to identify a marker of undifferentiated ES/primordial germ (PG) cells. In RT-PCR analysis, embryonic (E)-AP was detected only in undifferentiated ES cells, but intestinal(I)-AP was not detected in all of the examined ES and PG cells. In addition, nonspecific (NS)-AP wasdetected in undifferentiated PG cell from day 7, 5 to 13 of gestation. Histological activity of AP in ES cells was completely suppressed by addition of L-phenylalanine (Phe), L-homoarginine (Har), and L-phenylalanylglycylglycine (PheGlyGly) as an inhibitor, but RT-PCR showed the same results as in the absence of an inhibitors. Our findings suggested that expression of E-AP and NS-AP may use as a marker to determine the undifferentiated status in ES and PG cells.

• Bulletin of the Korean Chemical Society
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• v.29 no.5
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• pp.943-947
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• 2008

A phosphate-specific and fluorescent probe was prepared for label-free phosphatase assays based on fluorescence polarization. By using the probe, dephosphorylation reactions of DNA and protein substrates by calf intestinal alkaline phosphatase (CIP) could effectively be monitored in real-time. Since this assay method does not require additional materials such as labeled substrates and phosphospecific antibodies to obtain fluorescence polarization signals, it is simple, cost-effective, and expected to be useful not only for measuring activity of phosphatases but also for high-throughput screening of phosphatase inhibitors.

• The korean journal of orthodontics
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• v.30 no.2 s.79
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• pp.205-214
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• 2000

Recently, the magnetic force has been considered as a method for a more efficient tooth movement. The purpose of this study was to evaluate the effects of different static magnetic fields of Nd-Fe-B magnet on MC3T3-E1 cells by measuring the alkaline phosphatase activity and observing the amount of stained alkaline phosphatase. For measuring of alkaline phosphatase activity, MC3T3-E1 cells were seeded in first and third row of 12 well culture plates. And Nd-Fe-B magnets were positioned under the first column of first and third row to apply different static magnetic fields(first column:100mT ; second column:4.6mT ; third column:0.5mT ; forth column:0.0mT) to the cells for 7, 13, 19, and 25 days. For staining of alkaline phosphatase, MC3T3-E1 cells were seeded in 100mm culture plates. And Nd-Fe-B magnets were positioned under the corner of plates to apply different static magnetic fields(magnet side:100mT : the opposite side:0.5mT) to the cells for 7, 13, 19, and 25 days. The results were as follows : 1. ALP activity was increased until day 19 in biochemical determination as well as in histochemical staining, 2. The application of higher magnetic field(100mT) suppressed ALP activity at day 13, 19, 25. On the contrary, the application of the lower magnetic field(4.6mT, 0.5mT) significantly enhanced the ALP activity. 3. Consistent with enzyme assay, histochemical staining of ALP also demonstrated that higher magnetic field(100mT) suppressed ALP activity, lower one(0.5mT) enhanced.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.10-17
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• 2016

Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.

• The korean journal of orthodontics
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• v.25 no.6 s.53
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• pp.697-704
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• 1995

Chicken calvarial bone is known to contain various cell types, but their exact composition is unknown. By characterizing the chicken calvarial bone biochemically, it can be used to study biochemical, histochemical actions of bone cells in general. Calvaria of 18-day-old white leg horn embryo was aseptically dissected and bone cell populations were isolated by sequential enzymatic digestion. Histochemical study for osteoclast-like bone cell. population was performed with tartrate resistant acid phosphatase(TRAP) stain and for osteoblast-like bone cell population, alkaline phosphatase(ALP) stain was performed. Biochemical study for osteoblast-like bone cell population was performed using alkaline phosphatase(ALP) assay. Following conclusions were obtained from this study. 1. TRAP positive multi and mononuclear cells were mostly observed in group I and II, indicating that osteoclast-like bone cell population is mostly found in these groups. 2. All the cultured groups showed almost equal ALP activities and were positive for ALP stain, indicating that osteoblast-like bone cell population is evenly dispersed in all culture groups. 3. Experimental group treated with $1,25(OH)_{2}D_3$ showed increase in ALP activity in contrast to the control group, confirming previous studies that $1,25(OH)_{2}D_3$ increases ALP activities in in vitro bone cultures. 4. Results from von Kossa's stain indicated that in vitro bone formation had occured after 3 weeks of culture with beta-glycero phosphate.

• BMB Reports
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• v.41 no.4
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• pp.305-309
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• 2008

Cobalt is an essential microelements in many biological processes involving enzymatic activity. We found that $Zn^{2+}$ and $Mg^{2+}$, which are in the active site of native calf intestine alkaline phosphatase (CIP), can be replaced by $Co^{2+}$ directly in solution. The effect of $Co^{2+}$ concentration on the substitution reaction was examined at ratios of [$Co^{2+}$]/[CIP] from 0:1 to 8:1. The quantity of $Zn^{2+}$ in CIP decreased progressively as the ratio was increased, but the amount of $Mg^{2+}$ changed in irrregular fashion. A series of active site models of the reaction mechanism of CIP are proposed. Low pH was found to promote the replacement of $Mg^{2+}$ by $Co^{2+}$. To understand how the substitution affects the enzyme, we also solved the secondary structure of CIP after reaction with $Co^{2+}$ in different conditions.