• Title/Summary/Keyword: alkaline phosphate

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Effect of Fibroblast Growth Factor 23 on Osteoblastic Differentiation and Mineralization of D1 Mesenchymal Stem Cells (섬유모세포성장인자-23이 D1 간엽줄기세포에서 조골세포로의 분화 및 기질 광화에 미치는 영향)

  • Park, Kyeong-Lok
    • Journal of Life Science
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    • v.26 no.3
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    • pp.331-337
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    • 2016
  • Although fibroblast growth factor 23 (FGF23) is exclusively produced in osteoblasts and osteocytes, its main target is the kidney, where it decreases phosphate reabsorption by suppressing Na-phosphate cotransporters. Independently of its action on phosphate homeostasis, FGF23 also inhibits bone formation in vivo. In a calvarial osteoblastic cell model, FGF23 was shown to negatively affect extracellular matrix mineralization. This study investigated whether FGF23 had similar effects on osteoblast maturation, including differentiation and mineralization of bone marrow-derived mesenchymal stem cells (MSCs). D1 MSCs were cultured in an osteogenic medium containing β-glycerophosphate, ascorbic acid, and dexamethazone. Osteoblastic differentiation was evaluated by alkaline phosphatase (Alp) staining, and matrix mineralization was evaluated by alizarin red staining and calcium deposition. The expression of differentiation-stimulating genes Runx2, Alp, and osteocalcin and mineralization-inhibiting genes Enpp1 and Ank was analyzed using semiquantitative RT-PCR. Supraphysiological doses of FGF23 did not stimulate proliferation or osteoblastic differentiation of MSCs. Matrix mineralization 1, 2, and 3 weeks after the FGF23 treatment did not vary between control and FGF23 groups, although time-dependent enhancement of mineralization was obvious. Calcium deposition was also unchanged after the FGF23 treatment. mRNA expression levels of differentiation- and mineralization-related genes were also similar between the groups. Despite these negative findings, FGF23 signaling through FGF receptors seemed to function normally, with phosphorylation of the Erk protein more evident in the FGF23 group than in controls. These findings suggest that unlike calvarial osteoblasts, FGF23 is not likely to affect osteoblastic differentiation and mineralization of MSCs.

Transcriptional regulation of soybean ${\beta}-conglycinin$ gene expression: -(II) Developmental change of soybean embryo factor 3 activity- (대두 ${\beta}-conglycinin$ 유전자 발현의 전사 조절에 관한 연구 -(II) 대두 발달과정 중의 대두 배 인자 3의 역가 변화-)

  • Lee, Kyung-Hoon;Chung, Dong-Hyo;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.553-556
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    • 1993
  • Soybean nuclear extracts were prepared to examine the expression of SEF3 (soybean embryo factors 3), which binds to the upstream region of soybean ${\beta}-conglycinin$ ${\alpha}'$ subunit gene and is presumed to be a trans-acting factor for the expression of the gene. The relative levels of SEF3 binding activity in nuclear extracts of maturing soybean embryos were determined using the SE3 DNA probe containing two AACCCA hexanucleotides for gel mobility shift assay. The SEF3 activity increased in developing embryos from 16 to 32 days after pollination, whereas the mobility of the SE3-SE3-SEF3 complex decreased. The mobility of the complex was increased by the treatment of nuclear extracts with alkaline phosphatese, which could be inhibited by phosphate. Formation of the SE3-SEF3 complex was not affected by the binding buffer pH between 6.8 and 8.5.

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A Cell-Based Assay System for Monitoring NF-$\kappa$B Activity in Human Epidermal Keratinocytes: A Screening Tool of the Antioxidants and Anti-inflammatories for Dermatological Purpose

  • Moon, Ki-Young;Hahn, Bum-Soo;Lee, Jinseon;Kim, Yeong-Shik
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.17-27
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    • 2001
  • A cell-based assay system for monitoring NF-$textsc{k}$B activity was developed to determine the influence of activated NF-$textsc{k}$B in human HaCaT cells. The pNF-$textsc{k}$B-SEAP-NPT plasmid that permits expression of the secreted alkaline phosphatase (SEAP) reported gene in response to the NF-$textsc{k}$B activity and contains neomycin phosphotransferase (NPT) gene for the geneticin resistance in host cells was constructed and transfected into human keratinocyte cell line HaCaT. Human HaCaT transfectant cells secreted the SEAP enzyme into the culture medium in a time-dependent manner until 72h. NF-$textsc{k}$B activities were measured in the SEAP reporter gene assay using a fluorescent detection method. The treatment of HaCaT cell transfectants with known antioxidants [e.g., N-acetyl-L-cysteine and vitamin C] showed inhibition of NF-$textsc{k}$B activity in a time-and concentration-dependent manner. The phorbol 12-myristate 13-acetate (PMA) known as a stimulator of NF-$textsc{k}$B expression demonstrated that it increased NF-$textsc{k}$B activity in a time- and concentration-dependent manner. This assay system could be used to determine the quantitative measurement of NF-$textsc{k}$B activity in the human skin and allow the screening of anti-inflammatory agents from various synthetic chemicals and natural products for dermatological purpose. Abbrevitions used: NF-$textsc{k}$B, nuclear factor kappa B; I-$textsc{k}$B, Inhibitory kappa B; SEAP, secreted alkaline phosphatase; NPT, neomycin phosphotransferease; PCR, polymerase chain reaction: dNTP, deoxynucleoside triphosphates; DMEM, dulbecco’s modified eagle medium; FBS, fetal bovine serum; PBs, phosphate-buffered saline; MUP, 4-methylumbellifery phosphate; NAC, N-acetyl-L-cysteine; DMSO, dimethyl sulfoxide; PMA, phorbol 12-myristate 13-acetate.

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Age-related Changes in blood Chemistry and Thyroid hormones in Senescence Accelerated Mice(SAM R1 and SAM P6) (노화 방지를 위한 한약재의 효능 연구 (I))

  • Kim, Chung-Sook;Lee, Je-Hyun;Ma, Jin-Yeul;Jeon, Won-Kyung
    • 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.

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Isolation and Identification of Alkali-tolerant Bacteria from Near-Shore Soils in Dokdo Island

  • Namirimu, Teddy;Kim, Jinnam;Zo, Young-Gun
    • Microbiology and Biotechnology Letters
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    • v.47 no.1
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    • pp.105-115
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    • 2019
  • Saline or alkaline condition in soil inhibits growth of most crop plants and limits crop yields in many parts of the world. Augmenting an alkaline soil with alkali-tolerant bacteria capable of promoting plant growth can be a promising approach in expanding fertile agricultural land. Near-shore environments of Dokdo Island, a remote island located in the middle of the East Sea, appear to have patches of seawater-influenced haloalkaline soil that is unsupportive for growth of conventional plants. To exploit metabolic capacities of alkali-tolerant bacteria for promoting plant growth in saline or alkaline soils, we isolated of alkali-tolerant bacteria from near-shore soil samples in Dokdo and investigated properties of the isolates. Alkali-tolerant bacteria were selectively cultivated by inoculating suspended and diluted soil samples on a plate medium adjusted to pH 10. Fifty colonies were identified based on their $GTG_5$-PCR genomic fingerprints and 16S rRNA gene sequences. Most isolates were affiliated to alkali-tolerant and/or halotolerant genera or species of the phyla Firmicutes (68%), Proteobacteria (30%) and Actinobacteria (2%). Unlike the typical soil bacterial flora in the island, alkali-tolerant isolates belonged to only certain taxa of terrestrial origin under the three phyla, which have traits of plant growth promoting activities including detoxification, phytohormone production, disease/pest control, nitrogen-fixation, phosphate solubilization or siderophore production. However, Firmicutes of marine origin generally dominated the alkali-tolerant community. Results of this study suggest that haloalkaline environments like Dokdo shore soils are important sources for plant growth promoting bacteria that can be employed in bio-augmentation of vegetation-poor alkaline soils.

Phosphate sorption to quintinite in aqueous solutions: Kinetic, thermodynamic and equilibrium analyses

  • Kim, Jae-Hyun;Park, Jeong-Ann;Kang, Jin-Kyu;Kim, Song-Bae;Lee, Chang-Gu;Lee, Sang-Hyup;Choi, Jae-Woo
    • Environmental Engineering Research
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    • v.20 no.1
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    • pp.73-78
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    • 2015
  • The aim of this study was to examine the phosphate (P) removal by quintinite from aqueous solutions. Batch experiments were performed to examine the effects of reaction time, temperature, initial phosphate concentration, initial solution pH and stream water on the phosphate adsorption to quintinite. Kinetic, thermodynamic and equilibrium isotherm models were used to analyze the experimental data. Results showed that the maximum P adsorption capacity was 4.77 mgP/g under given conditions (initial P concentration = 2-20 mgP/L; adsorbent dose = 1.2 g/L; reaction time = 4 hr). Kinetic model analysis showed that the pseudo second-order model was the most suitable for describing the kinetic data. Thermodynamic analysis indicated that phosphate sorption to quintinite increased with increasing temperature from 15 to $45^{\circ}C$, indicating the spontaneous and endothermic nature of sorption process (${\Delta}H^0=487.08\;kJ/mol$; ${\Delta}S^0=1,696.12\;J/(K{\cdot}mol)$; ${\Delta}G^0=-1.67$ to -52.56 kJ/mol). Equilibrium isotherm analysis demonstrated that both Freundlich and Redlich-Peterson models were suitable for describing the equilibrium data. In the pH experiments, the phosphate adsorption to quintinite was not varied at pH 3.0-7.1 (1.50-1.55 mgP/g) but decreased considerably at a highly alkaline solution (0.70 mgP/g at pH 11.0). Results also indicated that under given conditions (initial P concentration=2 mgP/L; adsorbent dose=0.8 g/L; reaction time=4 hr), phosphate removal in the stream water (1.88 mgP/g) was lower than that in the synthetic solution (2.07 mgP/g), possibly due to the presence of anions such as (bi)carbonate and sulfate in the stream water.

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.

소양호에서의 유기인산염 분해율

  • 최승익;안태석;김범철
    • Korean Journal of Microbiology
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    • v.30 no.2
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    • pp.113-118
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    • 1992
  • To elucidate a part of phosphorus cycle an'd the nutritional status of phosphorus. the degradation rates of organic phosphate by phosphatase activity (PA). were studied at the water column of Lake Soyang. from March 1900 to April 1991. Phosphatase activity showed the range of 1-2220 nMillhr. Its maximum value was recortled on August and minimum during October and November. The PA and chlorophyll u v~luess howed high correlation coefficent (0.69). and the values of specific astivity was highest during Winter, 45-12\ulcorner nMihripg chl. a, and lowest on October. 2 nMl hripg chl. 11. The values of PA and bioavailable dissolved inorganic phosphate (DIP) showed negative correlation in surface water. During destratification period (Oct. and Nov. 1990). the values of PA were about 11240 times lower than those during August albeit high concentration of chlorophyll a (1.7- 7.2 mglm'). Such results seem to be cause'd by DIP supply from the metalimnion. hypolimnion ant1 sediment. With these results. phosphate was sufficient to sustain the biomass in Autumn. and internal loading of phosphate shoultl accelerate the eutrophication in Lake Soyang.

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Phosphorus Release from Waste Activated Sludge by Microwave Heating (마이크로웨이브를 이용한 잉여 슬러지 가온과 인산염 방출)

  • Ahn, Johwan;Yang, Hoiweon;Kim, Jangho;Min, Sungjae;Kim, Junghwan
    • Journal of Korean Society on Water Environment
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    • v.33 no.4
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    • pp.387-393
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    • 2017
  • A chemical batch tests were conducted to evaluate if microwave heating enhances phosphorous release from waste activated sludge (WAS) at pH 2.5, 5, 7, 9 and 11. Polyphosphate-accumulating organisms have a unique physiological feature, which releases intracellular polyphosphate granules when they are exposed under high temperature environments. Microwave irradiation was found to encourage large amount of phosphorus release from WAS, depending on pH and temperature conditions. Most of phosphorus was released below $59^{\circ}C$ within 30 min. A marked increase in phosphorus release was observed under alkaline or acidic conditions. However, based on control tests for phosphorus release under different pH conditions without microwave heating, the largest amount of phosphorus released by microwave irradiation was found at pH 7, followed by 5, 9, 11. On the other hand, crystallization was conducted to obtain magnesium ammonium phosphate (MAP) from phosphate released by microwave heating at pH 7. X-ray diffraction analysis confirmed that the recovered crystalline materials were MAP. MAP is an environmentally friendly fertilizer, which slowly releases ammonia and phosphorus in response to the demand of plant root. Thus, the recovered MAP as a phosphate fertilizer is fully expected to play a important role in the reduction of agricultural non-point pollution.

Drug-Biomacromolecule Interaction VIII

  • Kim, Chong-Kook;Yang, Ji-Sun;Lim, Yun-Su
    • Archives of Pharmacal Research
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    • v.7 no.2
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    • pp.87-93
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    • 1984
  • The effects of ionic strength and pH on the binding of cefazolin to bovine serum albumin (BSA) were studied by UV difference spectrophotometry. As ionic strength at constant pH and temperature increases, the apparent bining constant decreased but the number of binding sites remained almost constant at 2. The constancy of the number of binding sites with increasing the ionic strength suggests that purely electrostatic forces between BSA and drug do not have great importance in the drug binding, even though there is a decrease in the apparent binding constant. Thus, the effect of ionic strength on the interaction between drug and BSA may be explained by the changes in ionic atmosphere of the aggregated BSA molecules and competitive inhibition by phosphate ions. In addition, the higher apparent binding constant at high ionic strength is explained by conformational changes of BSA from its aggregate forms into subunits. The pH effects on the afinity of interactions indicated that the binding affinity of cefazoline is higher in the neutral region than in the alkaline region. An d at high pH value, the number of binding sites decreased from 2 to 1 because of the conformational change of BSA in the alkaline region.

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