• Title/Summary/Keyword: phosphatase 활성도

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An Analysis for Effects of Stain Family Drugs on Osteogenic Differentiation using Human Periosteum-derived Mesenchymal Stem Cells (스타틴(statin) 약물이 성체줄기세포의 골분화에 미치는 영향)

  • Moon, Dong Kyu;Yun, Jeong-Won;Kim, Bo Gyu;Lee, A Ram;Moon, Sun Young;Byun, June-Ho;Hwang, Sun-Chul;Woo, Dong Kyun
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1337-1344
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    • 2019
  • Osteoporosis is characterized by a reduction in bone mass and typically manifests as an increase in fractures. Because this disease is common in elderly populations and lifespans are rapidly increasing, the incidence of osteoporosis has also grown. Most drugs currently used for osteoporosis treatment target osteoclasts in the bone tissue to prevent absorption. However, these medications also cause certain side effects and, furthermore, cannot increase bone mass. Thus, in order to control osteoporosis, regenerative medicine that utilizes adult stem cells and osteoblasts has been extensively studied. Statins, also known as 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors, are cholesterol-lowering drugs that have been widely prescribed for cardiovascular diseases. Interestingly, recent studies have reported the beneficial effects of various statins on bone formation via the activation of osteoblasts. Thus, the current study investigated the effects of seven statin-family drugs on osteoblast activity during osteogenic differentiation using adult stem cells from human periosteal tissue. Specifically, statin effects on alkaline phosphatase activity, an early marker of bone cell differentiation, and on calcium deposit, a late marker of bone cell differentiation, were assessed. The results demonstrate that some statins (for example, pitavastatin and pravastatin) have a weak but positive effect on bone formation, and the findings therefore suggest that statin treatments can be a novel modulator for osteogenic differentiation and regenerative medicine using periosteal stem cells.

Effects of Feeding the Mixed Oils of Butter, Sardine and Safflower on the Lipid Components in Serum and Activities of Hepatic Functional Enzymes in Rats (버터, 정어리유 및 홍화유의 혼합 급이가 흰쥐의 혈청 지질성분 및 간기능 효소 활성에 미치는 영향)

  • Kim, Han-Soo;Chung, Seung-Yong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.21 no.6
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    • pp.608-616
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    • 1992
  • This study was designed to observe the effects of feeding the mixed oils of the sardine oil containing n-3 EPA, DHA and the safflower oil which is rich in n-6 linoleic acid on the improvement of the lipids and enzyme activities of serum in rats. Experimental oils mixed with 16% butter (control group) and 8% butter + 8% olive oil, 8% butter and various level of sardine and safflower oils were administered to the male rats of the Sprague Dawley for 4 weeks. The activities of aspartate aminotransferase (AST, EC 2.6.1.1), alanine aminotransferase (ALT, EC 2.6.1.2), lactate dehydrogenase (LDH, EC 1.1.1.27) and alkaline phosphatase (ALP, EC 3.1.3.1) in serum were significantly decreased in the all experimental groups than in the control groups, and activities of ALT and LDH were remarkably lower in the group 5 (4% sardine 0il + 4% safflower oil). Concentrations of total cholesterol and HDL-cholesterol in serum were lower in the other groups than in the dontrol groups, and particularly, lowest in the group 5. Concentrations of LDL, LDL-cholesterol, phospholipid and triglyceride in serum were lower in the all experimental groups than in the control group. Concentrations to total cholesterol and cholesteryl ester in serum were lowest in the group 5. The ratio of cholesteryl ester to total cholesterol was remarkably high in the control group, while group 2 (8% olive oil) was the lowest. From this results, the feeding equal quantity mixed oil with n-3 PUFA rich sardine oil and n-6 PUFA rich safflower oil were effective on the improvement of the lipid composition in the serum. It might be due to the effects of appropriate ratios of P/S, 0.85 and n-6/n-3P, 2.85 in the test lipids.

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EFFECTS OF $INTERFERON-\gamma$ ON COLLAGEN AND FIBRONECTIN SYNTHESIS IN PRIMARY CULTURED PERIODONTAL LIGAMENT CELLS ($Interferon-\gamma$가 치주인대 세포의 Collagen 및 Fibronectin의 합성과 Alkaline Phosphatase 활성에 미치는 영향)

  • Kim, Gwang-Seok;Sung, Jae-Hyun;Choi, Je-Yong;Ryou, Hyun-Mo
    • The korean journal of orthodontics
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    • v.23 no.2 s.41
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    • pp.229-248
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    • 1993
  • [ $Interferon-\gamma$ ] has been suggested as a cytokine of connective tissue stabilizer. In addition, it has also been demonstrated that this cytokine inhibited bone remodeling activities of the bone derived cells. In order to illuminate the effects of this cytokine in orthodontic force induced bone remodeling, it was administered to primary cultured periodontal ligament cells which have been known to have some osteoblast like characteristics. $Interferon-\gamma$ slightly decreased $[^3H]thymidine$ incorporation rate without a significant change in the total cellular DNA content up to 1000 U/ml, which meant these doses were not cytotoxic to the cell. Total protein synthesis was not influenced by various concentration of interferon-y whether it was determined by the $[^3H]proline$ incorporation rate or by the Lowry smethod. The effect of $interferon-\gamma$ on the individual protein was, however, differential, ie, it increased $[^3H]proline$ incorporation into the noncollagenous protein marginally, while it decreased $[^3H]proline$ incorporation into the collagen, so that it caused dose-dependent suppression of the relative collagen synthesis. On the contrary, the fibronectin synthesis determined by the ELISA was increased by 1000 U/ml of $interferon-\gamma$. The differential effects of the interferon-y on the collagen and fibronectin synthesis exhibited not only their protein level but also the steady state mRNA level. $Interferon-\gamma$ decreased steady state level of ${\alpha}1(I)$ procollagen mRNA significantly, while showing no significant changes in the fibronectin mRNA level. In addition to this, it was also found that indomethacin did not affect on the $interferon-\gamma$ induced collagen decrease in this cell, which meant prostaglandins were not involed in the process of $interferon-\gamma$ induced collagen decrease. So it can be concluded that the incubation of periodontal ligament cells with 1000 U/ml of $interferon-\gamma$ for 24 hr showed differential effects on the type I collagen and fibronectin gene expression. The decrease in relative collagen synthesis in the protein level was related with decrease in the steady state level of mRNA, while the increase in the fibronectin synthesis in the protein level was not correlated with the mRNA level.

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Serum Enzyme and Isozyme Activities of Rats Acclimated to Cold Environment (寒冷環境에 순화시킨 흰쥐의 血淸酵素 및 同位酵素의 活性)

  • 정애순;남상열
    • The Korean Journal of Zoology
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    • v.29 no.2
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    • pp.107-120
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    • 1986
  • The activities of serum of serum lactate dehydrogenase (SLDH), serum alkaline phosphatase (SALP), serum creatine phosphokinase (SCPK), and their isozymes were determined in adult male Sprague-Dawley rats acclimated to cold environment $(4\\pm1^\\circC)$ for 36 days. The SLDH activity was significantly higher in the early stage of acclimated period. The steady state of SLDH activity seemed to be reached by the end of acclimated period. Electrophoretic separation of serum of control rat showed three SLDH isozymes. Isozymes SLDH4 and SLDH5 appeared most prominently, whereas only trace of SLDH1 or SLDH2 was found. The increase in SLDH level during acclimation to cold environment is a reflection of an immediate increase in the SLDH1, SLDH2, and SLDH3 type of SLDH isozyme. The acclimation to cold environment increased significantly level of SALP in the early state of acclimated period. SALP activity showed a attaining steady state with the resting level after transient rise. Electrophoretic separation of SALP of control rats showed the SALP1 and SALP2 fractions. The transient rise in SALP activity of rats acclimated to cold environment coincided with a transient rise in SALP1 fraction. Immediately after exposure to cold environment, there was significant elevation in SCPK activity. Value returned to normal after transient rise. A new steady state of SCPK activity seemed to be reached by 36 days. It may be inferred from the above data that thermal compensation appears to result from a change in the activity of an enzyme and that the SLDH, SLDH-isozyme, SALP-isozyme, and SCPK may be involved directly or indirectly in thermoregulation during acclimation to cold environment.

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Phenotypic Characterization of Cementum-Derived Cells in Human (사람 백악질 유래 세포의 형질 특성에 관한 연구)

  • Kim, Su-Hwan;Yang, Byung-Kun;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Lee, Yong-Moo
    • Journal of Periodontal and Implant Science
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    • v.34 no.2
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    • pp.269-279
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    • 2004
  • 백악질 세포의 분리 및 배양방법을 확립하고, 이를 이용하여 백악질 세포의 형질특성을 알아보고자 하였다. 교정목적으로 발거된 소구치를 이용하여, 치은섬유아세포, 치주인대 세포 및 백악질 유래세포를 분리, 배양하였다. 백악질 유래 세포 배양시에는 백악질을 절제한 후 Collagenase P를 이용하여 백악질 유래 세포 외의 다른 세포의 개제를 배제하였고, 기질을 분해하여 세포의 분리 및 배양이 용이하도록 하였다. 분리 및 배양시기의 세포의 형태를 광화현미정을 이용하여 관찰하였다. 조골세포의 특성을 가지는 SaOs-2 세포를 대조군으로 이용하여 분리 및 배양된 세포군들을 동일한 조건으로 배양하였다. 3일 및 7일째에 세포증식도를 측정하였고 7일째에 ALPase 효소 활성도를 측정하였다. 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 osteopontin(OPN), Alkaline phosphatase(ALP), type I collagen(COL-I), Bone sialoprotein(BSP), BMP-2 및 osteocalcin(OC)의 발현을 비교 관찰하였다. 백악질 유래 세포의 분리 및 배양을 시도한 5명의 치아 중에서 3명의 치아에서 세포군을 배양해 낼 수 있었다. 배양한 백악질 유래 세포는 섬유아세포와 유사한 형태와 증식을 보였다. ALPase 효소 활성도 검사 결과 백악질 유래 세포는 SaOs-2 세포보다 낮은 활성도를 나타내었으며, 배양된 세포의 RT-PCR 결과 백악질 유래 세포군에서는 ALPase의 발현이 나타나지 않았고, 다른 조골세포 표식자의 발현도 낮게 나타났다. 이는 백악질 유래 세포가 조골세포 및 다른 대조군의 세포와는 다른 형질 특성을 가지고 있다는 것을 시사한다. 이상의 관찰결과로 사람의 백악질 유래 세포롤 백악질의 절제 및 효소처리 방법으로 효과적인 분리 및 배양이 가능하며, 이는 향후 백악질 세포의 형질 특성 및 백악질 형성의 분자적 기전을 파악하는 중요한 연구자료로 활용 될 수 있을 것으로 사료된다.

Membrane-associated Guanylate Kinase Inverted-3 Modulates Enterovirus Replication through AKT Signaling Activation (Membrane associated guanylate kinase inverted-3의 AKT signaling을 통한 enterovirus replication 조절)

  • Park, Jin-Ho;Namgung, Ye-Na;Lim, Byung-Kwan
    • Journal of Life Science
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    • v.26 no.10
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    • pp.1182-1188
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    • 2016
  • Membrane-associated guanylate kinase inverted-3 (MAGI-3) is a member of the family of membrane-associated guanylate kinases (MAGUKs). MAGI-3 modulates the kinase activity of protein kinase B (PKB)/AKT through interactions with phosphatase and tensin homolog (PTEN)/MMAC. Coxsackievirus B3 (CVB3) is a common causative agent of acute myocarditis and chronic dilated cardiomyopathy. Activation of AKT and extracellular signal-regulated kinases 1/2 (ERK1/2) is essential for CVB3 replication, but the relation between MAGI-3 signaling and CVB3 replication is not well understood. This study investigated the role of MAGI-3 in CVB3 infection and replication. MAGI-3 was overexpressed in HeLa cells by polyethylenimine (PEI) transfection. To optimize the transfection conditions, different ratios of plasmid DNA to PEI concentrations were used. MAGI-3 and empty plasmid DNA were transfected into the HeLa cells. MAGI-3 overexpression alone was not sufficient to efficiently activate AKT. However, expression of the CVB3 capsid protein VP1 dramatically increased in the HeLa cells overexpressing MAGI-3 24 h after CVB3 infection. In addition, the activities of AKT and ERK were significantly induced in the CVB3-infected MAGI-3 cells overexpressing HeLa. These results demonstrate that MAGI-3 expression upregulates CVB3 replication through AKT and ERK signaling activation. MAGI-3 may be an important target to control CVB3 replication.

Dynamics of Soil Microflora and Soil Enzymes around the Fairy-rings of Tricholoma matsutake (송이 균환(菌環) 주변(周邊)의 토양미생물(土壤微生物)과 토양효소(土壤酵素)의 동태(動態))

  • Hur, Tae Chul;Park, Hyun
    • Journal of Korean Society of Forest Science
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    • v.90 no.6
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    • pp.767-773
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    • 2001
  • We investigated the impacts of the fairy-ring of Tricholoma matsutake on the dynamics of soil microflora and soil enzyme activities by grouping the soils around the fairy-ring of T. matsutake into four regions. The regions were grouped as 'zone of decayed mycorrhizae', 'zone of mycorrhizae for fruiting', 'zone of physiologically active mycorrhizae' and 'zone free from mycorrhizal infection'. Soil fungi and actinomycetes were quite little at the soils around the fairy-ring of T. matsutake compared to those of general forest soils, and there were significant differences among the four regions. The soils with the mycelial cluster of T. matsutake showed about one third of microbial population compared to those in the zone free from mycorrhizal infection, which indicated that T. matsutake took a dominant position within the fairy-ring of the fungus. We could manifest that T. matsutake showed a distinctive characteristics of mycorrhizal fungus since the activities of dehydrogenase were significantly different between the zone of physiologically active mycorrhizae and the zone free from mycorrhizal infection. The dehydrogenase activity was the highest at the early season of fruiting around the fairy-ring of T. matsutake, while the acid-phosphatase activity increased from March to June followed by a slight decrease on August and peaked on October. This phenomenon made us infer that the vitality of T. matsutake be sustained after fruiting.

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Effects of Bisphosphonates on the Proliferation and the AlkalinePhosphatase Activity of Human Bone Marrow Derived Mesenchymal Stem Cells (Bisphosphonate가 인간 골수유래 간엽줄기세포의 증식과 알칼리성 인산분해효소 활성에 미치는 영향)

  • Jung, Jun-Ho;Lee, Baek-Soo;Kwon, Yong-Dae;Ohe, Joo-Young;Kim, Young-Ran
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.33 no.1
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    • pp.10-18
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    • 2011
  • Purpose: The purpose of this study is to find out the effects of bisphosphonates (BPs) on the proliferation and the alkaline phosphatase (ALP) activity of human bone marrow derived mesenchymal stem cells (hMSCs), and thus state its correlation with bisphosphonate related osteonecrosis of the jaw (BRONJ). Methods: hMSCs was obtained by collecting and culturing cancellous bone fragments from a patient undergoing iliac bone graft. Alendronate (Aln) and Pamidronate (Pam), Ibandronate (Ibn) were added to the culture media in the concentration from $10^{-3}$ M to $10^{-11}$ M and cell toxicity, viability were measured. For ALP activity evaluation, Aln and Pam were added to the culture media in the concentration from $5{\times}10^{-7}$ M to $1{\times}10^{-8}$ M and were cultured for 1 week, 2 weeks and 3 weeks. ALP activity data were standardized using protein assay. Control groups were prepared for each examination. Results: Aln, Pam and Ibn all failed to increase the proliferation of hMSCs. With 1 week, 2 weeks of $5{\times}10^{-8}$M of Aln treatment, the ALP activity increased. Pam treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and$1{\times}10^{-8}$M. Also Ibn treatment increased the ALP activity with 2 weeks of $5{\times}10^{-8}$ M and $1{\times}10^{-8}$ M. Conclusion: It is considered that BPs are not capable of improving the proliferation of hMSCs. Also, after a transient increase in the ALP activity with the lower concentration of BPs, the activity decreased again. Therefore, in patients on long-term medication of BPs, the proliferation and osteoblast differentiation of hMSCs are restrained, and thus delayed wound healing and increase in BRONJ complications may occur.

Pasteurization Efficiency of a Continuous Microwave HTST (연속식 마이크로파 고온단시간 살균시스템에서 우유의 살균효과)

  • Kim, Suk-Shin;Lee, Joo-Hee;Kim, Sang-Yong
    • Korean Journal of Food Science and Technology
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    • v.31 no.5
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    • pp.1392-1396
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    • 1999
  • This work was to apply the microwave energy to HTST pasteurization of milk in order to prevent undesirable quality changes due to the fouling and overheating on the surface of heat exchanger. A continuous tubulartype microwave pasteurization system was designed using a domestic microwave oven(800w and 2,450MHz). Raw milk was HTST pasteurized$(at\;72^{circ}C\;for\;15\;sec)$ by three methods; by heating in a stainless steel tube immersed in a hot water bath(MP0), by heating in a microwave cavity to a desired temperature and then holding in a hot water bath(MP1) and by both heating and holding in a microwave cavity(MP2). The microbial quality based on the total plate count and Psychotrophic bacterial count was in the order MP0, MP2 and MP1 ; however, the quality difference was not significant(p<0.05) when the initial microbial numbers were involved in the statistical analysis. In addition, the three samples pasteurized by different methods showed the similar microbial quality based on the coliform count and phosphatase activity. The similar microbial quality of the three samples supports the potential use of microwave energy for the pasteurization of milk and other fluid food products.

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Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis (신생 생쥐 고환에서 기인한 다분화능 생식줄기세포주의 확립 및 특성 분석)

  • Han, Sang-Chul;Song, Haeng-Seok;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.1
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    • pp.39-48
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    • 2008
  • Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.