• Title/Summary/Keyword: bBMP

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Gonadotropins Improve Porcine Oocyte Maturation and Embryo Development through Regulation of Maternal Gene Expression

  • Wang, Qing-Ling;Zhao, Ming-Hui;Jin, Yong-Xun;Kim, Nam-Hyung;Cui, Xiang-Shun
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.361-371
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    • 2013
  • The present study assessed the effect of FSH and LH on oocyte meiotic, cytoplasmic maturation and on the expression level and polyadenylation status of several maternal genes. Cumulus-oocyte complexes were cultured in the presence of FSH, LH, or the combination of FSH and LH. Significant cumulus expansion and nuclear maturation was observed upon exposure to FSH alone and to the combination of FSH and LH. The combination of FSH and LH during entire IVM increased the mRNA level of four maternal genes, C-mos, Cyclin B1, Gdf9 and Bmp15, at 28 h. Supplemented with FSH or LH significantly enhanced the polyadenylation of Gdf9 and Bmp15; and altered the expression level of Gdf9 and Bmp15. Following parthenogenesis, the exposure of oocytes to combination of FSH and LH during IVM significantly increased cleavage rate, blastocyst formation rate and total cell number, and decreased apoptosis. In addition, FSH and LH down-regulated the autophagy gene Atg6 and upregulated the apoptosis gene Bcl-xL at the mRNA level in blastocysts. These data suggest that the FSH and LH enhance meiotic and cytoplasmic maturation, possibly through the regulation of maternal gene expression and polyadenylation. Overall, we show here that FSH and LH inhibit apoptosis and autophagy and improve parthenogenetic embryo competence and development.

Delivery of growth factor-associated genes to mesenchymal stem cells for cartilage and bone tissue regeneration

  • Ahn, Jongchan;Park, Seah;Cha, Byung-Hyun;Kim, Jae Hwan;Park, Hansoo;Joung, Yoon Ki;Han, Inbo;Lee, Soo-Hong
    • Biomaterials and Biomechanics in Bioengineering
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    • v.1 no.3
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    • pp.151-162
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    • 2014
  • Genetically-modified mesenchymal stem cells (GM-MSCs) have emerged as promising therapeutic tools for orthopedic degenerative diseases. GM-MSCs have been widely reported that they are able to increase bone and cartilage tissue regeneration not only by secreting transgene products such as growth factors in a long-term manner, also by inducing MSCs into tissue-specific cells. For example, MSCs modified with BMP-2 gene increased secretion of BMP-2 protein resulting in enhancement of bone regeneration, while MSCs with TGF-b gene did cartilage regeneration. In this review, we introduce several growth factors for gene delivery to MSCs and strategies for bone and cartilage tissue regeneration using GM-MSCs. Furthermore, we describe strategies for strengthening GM-MSCs to more intensively induce tissue regeneration by co-delivery system of multiple genes.

Discovery and characterization of berberine derivatives as stimulators of osteoblast differentiation

  • Han, Younho;Park, Won-Jong
    • International Journal of Oral Biology
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    • v.44 no.4
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    • pp.165-172
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    • 2019
  • Berberine has been used clinically for more than a decade to treat various diseases, has been shown to be effective in osteoblast differentiation, and is a potential treatment option for osteoporosis. However, compared with existing osteoporosis drugs, berberine is somewhat less effective. This study aimed to identify a new compound with efficacy superior to that of berberine. The osteogenic activities of various berberine derivatives were evaluated via cell differentiation in C2C12 preosteoblast cell lines. Alkaline phosphatase (ALP) staining assay and structure-activity relationship demonstrated that compound 2b had a potent osteogenic effect. Furthermore, compound 2b dose dependently increased ALP activity and showed no toxicity at the effective concentration, indicating its efficacy. Additionally, compound 2b upregulated BMP2-induced transcriptional activity in a promoter activity assay using ALP, BSP, and OC promoters.

Long Term Effect of High Glucose and Phosphate Levels on the OPG/RANK/RANKL/TRAIL System in the Progression of Vascular Calcification in rat Aortic Smooth Muscle Cells

  • Kang, Yang Ho;Jin, Jung Sook;Son, Seok Man
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.2
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    • pp.111-118
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    • 2015
  • Osteoprotegerin (OPG), receptor activator of NF-${\kappa}B$ ligand (RANKL)/receptor activator of NF-${\kappa}B$ (RANK) axis, and TNF-related apoptosis-inducing ligand (TRAIL) participate in vascular calcification process including atherosclerosis, but their contributions under high glucose (HG) and phosphate (HP) condition for a long-term period (more than 2 weeks) have not been fully determined. In this study, we evaluated the effects of HG and HP levels over 2 or 4 weeks on the progression of vascular calcification in rat vascular smooth muscle cells (VSMCs). Calcium deposition in VSMCs was increased in medium containing HG (30 mmol/L D-glucose) with ${\beta}$-glycerophosphate (${\beta}$-GP, 12 mmol/L) after 2 weeks and increased further after 4 weeks. OPG mRNA and protein expressions were unchanged in HG group with or without ${\beta}$-GP after 2 weeks. However, after 4 weeks, OPG mRNA and protein expressions were significantly lower in HG group with ${\beta}$-GP. No significant expression changes were observed in RANKL, RANK, or TRAIL during the experiment. After 4 weeks of treatment in HG group containing ${\beta}$-GP and rhBMP-7, an inhibitor of vascular calcification, OPG expressions were maintained. Furthermore, mRNA expression of alkaline phosphatase (ALP), a marker of vascular mineralization, was lower in the presence of rhBMP-7. These results suggest that low OPG levels after long term HG and phosphate stimulation might reduce the binding of OPG to RANKL and TRAIL, and these changes could increase osteo-inductive VSMC differentiation, especially vascular mineralization reflected by increased ALP activity during vascular calcification.

Ventx1.1 as a Direct Repressor of Early Neural Gene zic3 in Xenopus laevis

  • Umair, Zobia;Kumar, Shiv;Kim, Daniel H.;Rafiq, Khezina;Kumar, Vijay;Kim, SungChan;Park, Jae-Bong;Lee, Jae-Yong;Lee, Unjoo;Kim, Jaebong
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1061-1071
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    • 2018
  • From Xenopus embryo studies, the BMP4/Smad1-targeted gene circuit is a key signaling pathway for specifying the cell fate between the ectoderm and neuro-ectoderm as well as the ventral and dorsal mesoderm. In this context, several BMP4/Smad1 target transcriptional factors have been identified as repressors of the neuro-ectoderm. However, none of these direct target transcription factors in this pathway, including GATA1b, Msx1 and Ventx1.1 have yet been proven as direct repressors of early neuro-ectodermal gene expression. In order to demonstrate that Ventx1.1 is a direct repressor of neuro-ectoderm genes, a genome-wide Xenopus ChIP-Seq of Ventx1.1 was performed. In this study, we demonstrated that Ventx1.1 bound to the Ventx1.1 response cis-acting element 1 and 2 (VRE1 and VRE2) on the promoter for zic3, which is a key early neuro-ectoderm gene, and this Ventx1.1 binding led to repression of zic3 transcription. Site-directed mutagenesis of VRE1 and VRE2 within zic3 promoter completely abolished the repression caused by Ventx1.1. In addition, we found both the positive and negative regulation of zic3 promoter activity by FoxD5b and Xcad2, respectively, and that these occur through the VREs and via modulation of Ventx1.1 levels. Taken together, the results demonstrate that the BMP4/Smad1 target gene, Ventx1.1, is a direct repressor of neuro-ectodermal gene zic3 during early Xenopus embryogenesis.

Genetic Association of the Porcine C9 Complement Component with Hemolytic Complement Activity

  • Khoa, D.V.A.;Wimmers, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.9
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    • pp.1354-1361
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    • 2015
  • The complement system is a part of the natural immune regulation mechanism against invading pathogens. Complement activation from three different pathways (classical, lectin, and alternative) leads to the formation of C5-convertase, an enzyme for cleavage of C5 into C5a and C5b, followed by C6, C7, C8, and C9 in membrane attack complex. The C9 is the last complement component of the terminal lytic pathway, which plays an important role in lysis of the target cells depending on its self-polymerization to form transmembrane channels. To address the association of C9 with traits related to disease resistance, the complete porcine C9 cDNA was comparatively sequenced to detect single nucleotide polymorphisms (SNPs) in pigs of the breeds Hampshire (HS), Duroc (DU), Berlin miniature pig (BMP), German Landrace (LR), Pietrain (PIE), and Muong Khuong (Vietnamese potbelly pig). Genotyping was performed in 417 $F_2$ animals of a resource population (DUMI: $DU{\times}BMP$) that were vaccinated with Mycoplasma hyopneumoniae, Aujeszky diseases virus and porcine respiratory and reproductive syndrome virus at 6, 14 and 16 weeks of age, respectively. Two SNPs were detected within the third exon. One of them has an amino acid substitution. The European porcine breeds (LR and PIE) show higher allele frequency of these SNPs than Vietnamese porcine breed (MK). Association of the substitution SNP with hemolytic complement activity indicated statistically significant differences between genotypes in the classical pathway but not in the alternative pathway. The interactions between eight time points of measurement of complement activity before and after vaccinations and genotypes were significantly different. The difference in hemolytic complement activity in the both pathways depends on genotype, kind of vaccine, age and the interaction to the other complement components. These results promote the porcine C9 (pC9) as a candidate gene to improve general animal health in the future.

Melanin extract from Gallus gallus domesticus promotes proliferation and differentiation of osteoblastic MG-63 cells via bone morphogenetic protein-2 signaling

  • Yoo, Han-Seok;Chung, Kang-Hyun;Lee, Kwon-Jai;Kim, Dong-Hee;An, Jeung Hee
    • Nutrition Research and Practice
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    • v.11 no.3
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    • pp.190-197
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    • 2017
  • BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

Monitoring System of Agriculture Fields using ZigBee Modules

  • Ayurzana, Odgerel;Tsagaanchuluun, Sugir
    • International journal of advanced smart convergence
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    • v.10 no.1
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    • pp.89-96
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    • 2021
  • The goal of this study is to develop experiment monitoring system of agriculture fields using ZigBee wireless modules. Soil moisture, ambient temperature, atmospheric pressure and intensity of sunlight are the most important factorsto grow a wheat crop and other vegetables. In orderto monitorthe factorssoil moisture (YL69), air pressure (BMP180), temperature (DS18B20), photoresistor were used for sensing environment data. The TI CC2530 RF SoC chip was used in the system. ZigBee modules were connected to star topology. ZigBee modules send data wirelessly to a data center. This data can be displayed and analyzed on the main monitoring program as needed also sent to the client mobile. Characteristics of the sensors were determined by experiment results.

Biochemical Methane Potential of Agricultural Byproduct in Greenhouse Vegetable Crops (국내 주요 시설채소 부산물의 메탄 생산 퍼텐셜)

  • Shin, Kook-Sik;Kim, Chang-Hyun;Lee, Sang-Eun;Yoon, Young-Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.6
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    • pp.1252-1257
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    • 2011
  • Number of crop residues generated at large amount in agriculture can be utilized as substrate in methane production by anaerobic digestion. Greenhouse vegetable crop cultivation that adopting intensive agricultural system require the heating energy during winter season, meanwhile produce waste biomass source for the methane production. The purpose of this study was to investigate the methane production potential of greenhouse vegetable crop residues and to estimate material and energy yield in greenhouse system. Cucumber, tomato, and paprika as greenhouse vegetable crop were used in this study. Fallen fruit, leaf, and stem residues were collected at harvesting period from the farmhouses (Anseong, Gyeonggi, Korea) adopting an intensive greenhouse cultivation system. Also the amount of fallen vegetables and plant residues, and planting density of each vegetable crop were investigated. Chemical properties of vegetable waste biomass were determined, and theoretical methane potentials were calculated using Buswell's formula from the element analysis data. Also, BMP (Biochemical methane potential) assay was carried out for each vegetable waste biomass in mesophilic temperature ($38^{\circ}C$). Theoretical methane potential ($B_{th}$) and Ultimate methane potential ($B_u$) off stem, leaf, and fallen fruit in vegetable residues showed the range of $0.352{\sim}0.485Nm^3\;kg^{-1}VS_{added}$ and $0.136{\sim}0.354Nm^3\;kg^{-1}VS_{added}$ respectively. The biomass yields of residues of tomato, cucumber, and paprika were 28.3, 30.5, and $21.5Mg\;ha^{-1}$ respectively. The methane yields of tomato, cucumber, and paprika residues showed 645.0, 782.5, and $686.8Nm^3\;ha^{-1}$. Methane yield ($Nm^3\;ha^{-1}$) of crop residue may be highly influenced by biomass yield which is mainly affected by planting density.