• Title/Summary/Keyword: BMP2

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Kalkitoxin attenuates calcification of vascular smooth muscle cells via RUNX-2 signaling pathways

  • Saroj K Shrestha;Se-Woong Kim;Yunjo Soh
    • Journal of Veterinary Science
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    • v.24 no.5
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    • pp.69.1-69.11
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    • 2023
  • Background: Kalkitoxin (KT) is an active lipopeptide isolated from the cyanobacterium Lyngbya majuscula found in the bed of the coral reef. Although KT suppresses cell division and inflammation, KT's mechanism of action in vascular smooth muscle cells (VSMCs) is unidentified. Therefore, our main aim was to investigate the impact of KT on vascular calcification for the treatment of cardiovascular disease. Objectives: Using diverse calcification media, we studied the effect of KT on VSMC calcification and the underlying mechanism of this effect. Methods: VSMC was isolated from the 6 weeks ICR mice. Then VSMCs were treated with different concentrations of KT to check the cell viability. Alizarin red and von Kossa staining were carried out to examine the calcium deposition on VSMC. Thoracic aorta of 6 weeks mice were taken and treated with different concentrations of KT, and H and E staining was performed. Real-time polymerase chain reaction and western blot were performed to examine KT's effect on VSMC mineralization. Calcium deposition on VSMC was examined with a calcium deposition quantification kit. Results: Calcium deposition, Alizarin red, and von Kossa staining revealed that KT reduced inorganic phosphate-induced calcification phenotypes. KT also reduced Ca++-induced calcification by inhibiting genes that regulate osteoblast differentiation, such as runtrelated transcription factor 2 (RUNX-2), SMAD family member 4, osterix, collagen 1α, and osteopontin. Also, KT repressed Ca2+-induced bone morphogenetic protein 2, RUNX-2, collagen 1α, osteoprotegerin, and smooth muscle actin protein expression. Likewise, Alizarin red and von Kossa staining showed that KT markedly decreased the calcification of ex vivo ring formation in the mouse thoracic aorta. Conclusions: This experiment demonstrated that KT decreases vascular calcification and may be developed as a new therapeutic treatment for vascular calcification and arteriosclerosis.

EXPRESSION OF CARTILAGE DERIVED MORPHOGENIC PROTEIN IN PLEOMORPHIC ADENOMA (다형성선종에서 연골유래형성단백의 발현)

  • Lee, Hwang-Jae;Kim, Chul-Hwan;Kim, Kyung-Wook
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.3
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    • pp.247-255
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    • 2007
  • The bone morphogenic protein(BMP) can promote migration and growth of mesenchymal cells and initiate process for bone and cartilage formation. Cartilage-derived morphogenic protein(CDMP)-1 and -2 belong to the bone morphogenetic protein family in the transforming growth factor(TGF)-${\beta}$ superfamily. Although pleomorphic adenoma of the salivary glands is an epithelial tumor, it frequently shows ectopic cartilaginous formation with biomolecular studies. The mechanism of pathogenesis in cartilaginous formation is still controversy. We examined the expression and localization of CDMP-1 and -2, in comparison with the localization of cartilaginous matrix proteins, in human normal salivary glands and 20 cases of pleomorphic adenoma using immunohistochemical methods. The results were followed. 1. CMP-1 was immunolocalized in the striated ducts and the intercalated ducts, but not expressed in excretory duct, CDMP-2 was not expressed in the normal salivary glands. 2. CMP-1 was immunolocalized in the ductal cell and cuboidal neoplastic myoepithelial cells around the chondroid areas of the pleomorphic adenomas, whereas these molecules were not localized in the spindle-shaped neoplastic myoepithelial cells of the myxoid element in these tumors. CDMP-2 was expressed neither in normal salivary glands nor in any elements of the pleomorphic adenomas. 3. In transmission electron microscopic view, the tumor cells are composed of modifed myoepithelial cells between hyaline and myxoid stroma. 4. In Immuno-blot analysis, strong overexpression of CDMP-1 was frequently seen in pleomorphic adenomas, but the level of CDMP-2 was expressed minimally in pleomorphic adenoma. From the these results, it should be suggested that undifferentiated neoplastic myoepithelial cells around the chondroid areas expressed CDMP-1 and suggested that this molecule may play a role in the differentiation of neoplastic myoepithelial cells in pleomorphic adenoma, but not CDMP-2.

Investigation on the effect of water extracts of Mangifera indica leaves on the hair loss-related genes in human dermal papilla cells (망고 잎 열수 추출물의 모유두 세포에서 탈모 관련 유전자 발현에 미치는 영향 연구)

  • Choi, Youngsoo;Kim, Eunmi;Lee, Seong Hee;Han, Hyosang;Kim, Keekwang
    • The Korea Journal of Herbology
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    • v.36 no.3
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    • pp.39-46
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    • 2021
  • Objectives : Mangifera indica leaves are well known for having a variety of benefits, including anti-inflammatory, anti-tumor, diabetic retinopathy and diabetic vasculosis. However, the effects of Mangifera indica leaves on hair loss inhibition have not been studied. In this study, we investigated to find out the activity of Mangifera indica leaves on hair loss. Methods : 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid(ABTS) analysis was performed to confirm the antioxidant efficacy of the water extract of Mangifera indica leaves (WEML). To examine the effect of WEML on cell viability in dermal papillar (DP) cells, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetra Zolium (MTS) analysis was performed. The changes in the mRNA expression level of the hair loss and hair growth-related genes in dermal papilla cells by WEML treatment were confirmed by quantitative RT-PCR. Results : In dermal papilla (DP) cells, ABTS analysis and MTS analysis of WEML showed antioxidant efficacy and low cytotoxicity. As a result of gene expression analysis through Quantitative RT-PCR, no changes in hair growth-related genes BMP6 and CTNNB1 was confirmed. but inhibitory activity of WEML on hair loss-related genes EGR1, SGK, DKK1, SRD5A1 and SRD5A2 was confirmed. Conclusion : We confirmed that WEML has excellent antioxidant efficacy and a inhibitory activity of hair loss-related genes including 5α-reductase genes. These results suggest that Mangifera indica leaves have a potential activity as a hair loss treatment for hair loss and hair growth. Biochemical or molecular biological research on hair loss is needed.

Design of Structural BMPs for Low Impact Development (LID) Application and Modelling Its Effect on Reduction of Runoff and Nonpoint Source Pollution: Application of LIDMOD2 (저영향개발(LID)적용을 위한 구조적 BMPs의 유출량 및 비점오염저감 효과모의: LIDMOD2 적용)

  • Kim, Jung Jin;Kim, Tae-Dong;Choi, Donghyuk;Jeon, Ji-Hong
    • Journal of Korean Society on Water Environment
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    • v.27 no.5
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    • pp.580-586
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    • 2011
  • Low impact development (LID) technique is recently proposed as new concept to reduce surface runoff and pollutant loading with various best management practices (BMPs). In this study, LIDMOD2, which is one of the model to evaluate LID, was applied at Mohyeon developing area to evaluate the redcution of annual runoff and pollution loading, cost-reduction efficient by LID with design of structural BMPs including bioretention, wet pond, and wetland. As a simulation results, the bioretention had the highest reduction efficiency for runoff (41.43%), and 22% for T-N and 22% for BOD. Wet pont had the highest reduction efficiency for T-P as showing 25% of reduction rate. As a results of cost-reduction efficient, wet pont represented the highest cost-effective for T-N and BOD with showing 0.43 T-P kg/million won and 17.37 BOD kg/million won, respectively, and bioretention represented the highest cost-effective for T-P with showing 2.52 T-P kg/million won. LID technology could reduce effectively surface runoff and nonpoint source pollution and construct sustainable development. LIDMOD2 could be suggested as useful tool to evaluate and design LID.

Evaluation of the Biocompatibility of Cuttlebone in Mouse (쥐에서 오적골 생체적합성 평가)

  • Won, Sangcheol;Lee, Joo Myoung;Cheong, Jongtae;Park, Hyunjung;Seo, Jongpil
    • Journal of Veterinary Clinics
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    • v.32 no.5
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    • pp.417-421
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    • 2015
  • Bone grafting is widely used to bridge major bone defects or to promote bone union. Natural calcium carbonate (CC) has been used as a bone substitute material and used to scaffold for bone morphogenetic protein (BMP). The aims of this study is to evaluate the biocompatibility of cuttlebone (CB) and hydroxyapatite from CB (CBHA). Each material was shaped into disks (5 mm in diameter and 2 mm in thickness). To test biocompatibility, the disks were implanted into the dorsal subcutaneous tissue in mice. Fibrous capsule thickness around each disk was evaluated histologically at 2 and 4 weeks after implantation. Concerning biocompatibility, fibrous capsule thickness of CBHA was significantly thinner than that of CB and CHA (p < 0.05) at 2 and 4 weeks after implantation. Based on the clinical and histological results, CBHA would be a safe material for use inside the body and has more effective osteoconduction than CB.

The Experimental Studies of YangHyulEum Gami-Bang Extracts on the Hair Growth Effect (양혈음가미방(養血飮加味方) 추출물의 발모효과에 대한 실험적 연구)

  • Hong, Jee-Hee;Jung, Hyun-A
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.74-94
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    • 2016
  • Objectives : YangHyulEum Gami-Bang(YHEG) is a hair care extracts which is composed of fourteen plant extracts used in oriental medicine. The purpose of this study is to investigate the effect of YangHyulEum Gami-Bang(YHEG) on the alopecia and hair growth.Methods & Results : The herbal extracts from YangHyulEum Gami-Bang(YHEG) was tested using in vivo and in vitro test models. 1. The YHEG extracts showed effect on the DNA proliferation of the hair dermal papilla cells measured by [3H]thymidine incorporation. 2. YHEG showed promoting on the expression of growth factors such as IGF-1, KGF-1 and inhibiting on the expression of inhibitory hair growth factor such as TGF-β1, BMP-2 estimated by qPCR. 3. The YHEG extracts showed effect on the activation of β-catenin in the dermal papilla cells. 4. YHEG showed inhibitory effects of NO synthesis at 0.2% concentrations. 5. YHEG showed effects in the expression of IL-1β, TNF-α, IL-6, COX-2 and iNOS gene in the LPS stimulated RAW 264.7 cells. 6. The hair growth index of the YHEG extracts ranked at over 2 when compared to control group which was ranked at 0. 7. The hair follicle number, length and size of the experimental group were remarkably higher than the control group in the histological observation.Conclusions : These results suggest that YangHyulEum Gami-Bang(YHEG) has hair growth promoting activity and it can be used as a potent treatment agent for preventing hair loss and stimulating hair growth for treatment of alopecia.

Comparison of Non-Point Pollutant Loads by the Crops in Alpine Fields (고랭지 밭의 작물별 비점오염부하 비교)

  • Choi, Yong-Hun;Won, Chul-Hee;Kim, Tae-Yoo;Yang, Hee-Jeong;Choi, Joong-Dae
    • Proceedings of the Korea Water Resources Association Conference
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    • 2010.05a
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    • pp.1247-1252
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    • 2010
  • 강원도 평창군 대관령면 횡계리에 위치한 고랭지 밭을 2008년 3월부터 2009년 12월까지 발생한 강우사상에 대하여 모니터링을 실시하여, 경지단위의 밭에서 발생하는 비점오염부하를 측정하고, 작물에 의한 발생량을 비교하였다. 2008년에는 감자를 재배하였고, 2009년에는 무를 재배하였다. 2008년의 시비량은 2009년의 시비량보다 질소 2.1배, 인산 1.9배, 칼륨 2.3배 높게 나타났다. 2008년 총 5회의 유출에 의해 SS 2,908.47 kg/ha/yr, COD 67.95 kg/ha/yr, BOD 50.72 kg/ha/yr, TN 13.29 kg/ha/yr, TP 9.97 kg/ha/yr의 연 오염부하가 발생하였으며, 2009년 총 8회의 유출에 의해 SS 3,908.34 kg/ha/yr, COD 225.04 kg/ha/yr, BOD 156.96 kg/ha/yr, TN 18.88 kg/ha/yr, TP 36.41 kg/ha/yr의 연 오염부하가 발생하였다. 2008년과 2009년의 강우량이 다르기 때문에 연 오염 부하를 유출이 발생한 강우량의 합으로 나누어 비교하였다. 비료사용량이 2008년에 약 2배 많았음에도 불구하고 단위 강우량 당 오염부하에서 TN은 0.031 kg/ha/mm와 0.029 kg/ha/mm로 큰 차이가 나타나지 않았고, COD는 0.16 kg/ha/mm와 0.35 kg/ha/mm, BOD는 0.12 kg/ha/mm와 0.24 kg/ha/mm, TP는 0.023 kg/ha/mm와 0.057 kg/ha/mm로 2009년에 2배 이상 증가하였다. 이는 비료의 사용량보다 작물의 성장에 의한 지표피복효과가 비점오염 저감에 더 효과적 작용할 수 있다는 것을 의미한다. 따라서 고랭지 밭에서 피복율이 높은 작물을 재배하거나 피복재에 의한 지표피복을 실시하면 비점오염의 저감을 위한 최적관리방안으로 활용 될 수 있을 것이다.

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Methylation of the Mouse Dlx5 and Osx Gene Promoters Regulates Cell Type-specific Gene Expression

  • Lee, Ji Yun;Lee, Yu Mi;Kim, Mi Jin;Choi, Je Yong;Park, Eui Kyun;Kim, Shin Yoon;Lee, Sam Poong;Yang, Jae Sup;Kim, Dong Sun
    • Molecules and Cells
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    • v.22 no.2
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    • pp.182-188
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    • 2006
  • Dlx5 and Osx are master regulatory proteins essential for initiating the cascade leading to osteoblast differentiation in mammals, but the mechanism of osteoblast-specific expression is not fully understood. DNA methylation at CpG sequences is involved in tissue and cell type-specific gene expression. We investigated the methylation status of Dlx5 and Osx in osteogenic and nonosteogenic cell lines by methylationspecific PCR (MSP). The CpG dinucleotides of the Dlx5 and Osx promoter regions were unmethylated in osteogenic cell lines transcribing these genes but methylated in nonosteogenic cell lines. Treatment of C2C12 cells with 5-AzadC induced dose- and timedependent expression of Dlx5 and Osx mRNA by demethylating the corresponding promoters. Furthermore the mRNAs for the osteoblast markers ALP and OC, which were undetectable in untreated cells, gradually increased after 5-AzadC treatment. In addition, BMP-2 stimulation induced Dlx5 expression by hypomethylating its promoter. These findings suggest that DNA methylation plays an important role in cell type-specific expression of Dlx5 and Osx.

Growth and Osteoblastic Differentiation of Mesenchymal Stem Cells on Silk Scaffolds

  • Cho, Hee-Yeon;Baik, Young-Ae;Jeon, Suyeon;Kwak, Yoon-Hae;Kweon, Hae Yong;Jo, You Young;Lee, Kwang Gill;Park, Young Hwan;Kang, Dongchul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.27 no.2
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    • pp.303-311
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    • 2013
  • In this study, we compared the efficiency of osteoblast differentiation media (ODM) containing three distinct reagent combinations in osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramedullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be significantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentiation was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

The Bioenergy Conversion Characteristics of Feedlot Manure Discharging from Beef Cattle Barn

  • Oh, Seung-Yong;Kim, Chang-Hyun;Yoon, Young-Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.48 no.6
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    • pp.697-704
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    • 2015
  • This study was carried out to assess bioenergy conversion efficiency by biogas and solid fuel production in the cattle feedlot manure discharged from beef cattle barn. Feedlot manure was sampled from the cattle farmhouse located in Yong-in, Gyeonggi during the mid-fattening stage, periodically. The chemical characteristics, BMP (Biochemical methane potential) and HV (Heating values) of feedlot cattle manures were analyzed. Total solid contents of cattle feedlot manure were in the range of 29.98~44.28%, and volatile solid contents were in the range of 23.53~24.47%. In the anaerobic digestion of cattle feedlot manure, the methane production potential has increased from 0.141 to $0.187Nm^3kg^{-1}-VS_{added}$. The methane production of fresh cattle feedlot manure showed the range $0.141{\sim}0.187Nm^3kg^{-1}$-Manure (average $0.047Nm^3kg^{-1}$-Manure), the LHVs (lower heating values) of the produced methane were in the range of $316{\sim}560kcalkg^{-1}$-Manure (average $400kcalkg^{-1}$-Manure). In the direct combustion of fresh cattle feedlot manure, the LHVs were measured in the range of $747{\sim}1,271kcalkg^{-1}$-Manure (average $916kcalkg^{-1}$-Manure), and LHVs of solid fuel which have the water content of 20% were in the range of $2,694{\sim}2,876kcalkg^{-1}$-Manure (average $2,791kcalkg^{-1}$-Manure). Then, the drying energy of average $443kcalkg^{-1}$-Manure was consumed in the production of solid fuel which has a water content of 20%. Therefore, the direct combustion of cattle feedlot manure showed about 2.3 times higher LHV than the LHV of methane produced by anaerobic digestion. And LHV of solid fuel was about 6.0 times higher than the LHV of methane produced by anaerobic digestion. Then, the production of solid fuel presented more bioenergy conversion efficiency than the biogas production in the bioenergy use of cattle feedlot manure.