• Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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• 1998.11a
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• pp.236-239
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• 1998

This research was performed to check the transport characteristics of heavy metals in contaminated soil, that is, the influence of humic acid and phosphate on transport characteristics of heavy metals was studied. From the results of column mode experiments about heavy metal behavior, the order time to reach breakthrough and equilibrium was soil ＋ humic acid( 20g ) > soil ＋ humic acid ( 5 g ) > soil without Humic acid addition > soil+humic acid( 50g ). It is because the dissolved organic carbon content increased as the soil organic matter content increased. As the phosphate increased, so did the time to reach breakthrough and equilibrium. The order of time was soil + phosphate( 50 mg ) > soil + phosphate( 20 mg ) > soil . phosphate( 10 mg ) > soil without phosphate addition. It is because the phosphate ion worked as alkalinity donor and the calcium ion co-injected worked as the accelerator of coprecipitation of heavy metals.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.57-65
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• 2012

Phosphorous is an essential element for the synthesis of various biomolecules including phospholipids, carbohydrates and nucleic acids. Bacterial cells can uptake it as forms of phosphate and phosphate-containing nutrients from extracellular environments, and reserve extra phosphate to polyphosphate inside the cell. Among five phosphate transport systems, Pst plays central roles in phosphate transport, and its expression is coordinated by the regulation of PhoB-PhoR two component signal transduction system in response to extracellular levels of phosphate. Genomic studies on the response regulator PhoB reveal many genes independent of phosphate metabolism. Based on recent findings on phenotypes of bacteria lacking proper function of each phosphate transport system, this review discusses roles of phosphate transporters in maintaining optimum intracellular phosphate levels, and presents diverse phenotypes of phosphate transporters related with other environmental signals as well as phosphate, then finally points out functional redundancy among phosphate transport systems or their regulators, which emphasize importance of phosphate homeostasis in governing metabolism, adaptation, and virulence of bacteria.

• Journal of Acupuncture Research
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• v.19 no.4
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• pp.190-199
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• 2002

Objective : This study was performed to determine if Scutellaria balicalensis Georgi extract (SbGE) prevents oxidant-induced membrane transport dysfunction in renal tubular cells. Methods : Membrane transport function was estimated by measuring $Na^+$-dependent inorganic phosphate transport in opossum kidney (OK) cells. $H_2O_2$ inhibited phosphate transport in a dose-dependent manner. Results : The inhibitory effect of $H_2O_2$ was significantly prevented SbGE over concentration range of 0.005-0.05%. $H_2O_2$ caused ATP depletion, which was prevented by SbGE. $H_2O_2$ induced the loss of mitochondrial function as evidenced by decreased MTT reduction and its effect was prevented by SbGE. The $H_2O_2$-induced inhibition of phosphate transport was not affected by a potent antioxidant DPPD, but the inhibition was prevented by an iron chelator deferoxamine, suggesting that $H_2O_2$ inhibits $Na^+$-dependent phosphate transport via an iron-dependent nonperoxidative mechanism in renal tubular cells. Conclusion : These data suggest that SbGE may exert the protective effect against oxidant-induced membrane transport dysfunction by a mechanism similar to iron chelators in renal epithelial cells. However, furher studies should be carried out to find the active ingredient(s) of SbGE that exerts the protective effect.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.15-22
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• 2009

This study was undertaken to elucidate the underlying mechanisms of ATP depletion-induced membrane transport dysfunction and cell death in renal proximal tubular cells. ATP depletion was induced by incubating cells with 2.5 mM potassium cyanide(KCN)/0.1 mM iodoacetic acid(IAA), and membrane transport function and cell viability were evaluated by measuring $Na^+$-dependent phosphate uptake and trypan blue exclusion, respectively. ATP depletion resulted in a decrease in $Na^+$-dependent phosphate uptake and cell viability in a time-dependent manner. ATP depletion inhibited $Na^+$-dependent phosphate uptake in cells, when treated with 2 mM ouabain, a $Na^+$ pump-specific inhibitor, suggesting that ATP depletion impairs membrane transport functional integrity. Alterations in $Na^+$-dependent phosphate uptake and cell viability induced by ATP depletion were prevented by the hydrogen peroxide scavenger such as catalase and the hydroxyl radical scavengers(dimethylthiourea and thiourea), and amino acids(glycine and alanine). ATP depletion caused arachidonic acid release and increased mRNA levels of cytosolic phospholipase $A_2(cPLA_2)$. The ATP depletion-dependent arachidonic acid release was inhibited by $cPLA_2$ specific inhibitor $AACOCF_3$. ATP depletion-induced alterations in $Na^+$-dependent phosphate uptake and cell viability were prevented by $AACOCF_3$. Inhibition of $Na^+$-dependent phosphate uptake by ATP depletion was prevented by antipain and leupetin, serine/cysteine protease inhibitors, whereas ATP depletion-induced cell death was not altered by these agents. These results indicate that ATP depletion-induced alterations in membrane transport function and cell viability are due to reactive oxygen species generation and $cPLA_2$ activation in renal proximal tubular cells. In addition, the present data suggest that serine/cysteine proteases play an important role in membrane transport dysfunction, but not cell death, induced by ATP depletion.

• Journal of Plant Biology
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• v.21 no.1_4
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• pp.33-38
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• 1978

Phosphate absorption from a Na2H32PO4 solution by Oryza sativa L. was studied in order to elucidate kinetic mechanisms of ion transport. The rates of phosphate absorption from different concentraitons indicated the presence of dual mechanisms in root tips, one in the low (1$\times$10-6 to 8$\times$10-5M) and the other in the high (1$\times$10-4 to 8$\times$10-3M). A phosphate compensation point of phosphate transport was revealed with a 1$\times$10-6M solution of Na2H32PO4. The kinetic model that ion transport involves an exchange reaction of absorption and desorptin is prosposed as follows: where C represents an ionic-specific organic carrier in the membrane; M, Mo and Mi are the mineral ions, M-outside and M-inside; MC is a carrier-ion complex; and the K's represent rate constants. In this model, the Mi velocity, v, is given by: {{{{v= {dMi} over {dt}= {(K1K3Mo-K2K4Mi) Ct} over {(K2＋K3)＋K1Mo＋K4Mi} }} where Ct is equal to C＋MC, and t is time.

• The Korean Journal of Physiology
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• v.30 no.1
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• pp.63-76
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• 1996

The aim of present study was to characterize phosphate uptake and to investigate the mechanism for the insulin and insulin-like growth factor(IGF) stimulation of phosphate uptake in primary cultured rabbit renal proximal tubule cells. Results were as follows : 1. The primary cultured proximal tubule cells had accumulated $6.68{\pm}0.70$ nmole phosphate/mg protein in the presence of 140 mM NaCl and $2.07{\pm}0.17$ nmole phosphate/mg protein in the presence of 140 mM KCl during a 60 minute uptake period. Raising the concentration of extracellular phosphate to 100 mM$(48.33{\pm}1.76\;pmole/mg\;protein/min)$ induced decrease in phosphate uptake compared with that in control cells maintained in 1 mM phosphate$(190.66{\pm}13.01\;pmole/mg\;protein/min)$. Optimal phosphate uptake was observed at pH 6.5 in the presence of 140 mM NaCl. Phosphate uptake at pH 7.2 and pH 7.9 decreased to $83.06{\pm}5.75%\;and\;74.61{\pm}3.29%$ of that of pH 6.5, respectively. 2. Phosphate uptake was inhibited by iodoacetic acid(IAA) or valinomycin treatment $(62.41{\pm}4.40%\;and\;12.80{\pm}1.64%\;of\;that\;of\;control,\;respectively)$. When IAA and valinomycin were added together, phosphate uptake was inhibited to $8.04{\pm}0.61%$ of that of control. Phosphate uptake by the primary proximal tubule cells was significantly reduced by ouabain treatment$(80.27{\pm}6.96%\;of\;that\;of\;control)$. Inhibition of protein and/or RNA synthesis by either cycloheximide or actinomycin D markedly attenuated phosphate uptake. 3. Extracellular CAMP and phorbol 12-myristate 13 acetate(PMA) decreased phosphate uptake in a dose-dependent manner in all experimental conditions. Treatment of cells with pertussis toxin or cholera toxin inhibited phosphate uptake. cAMP concentration between $10^{-6}\;M\;and\;10^{-4}\;M$ significantly inhibited phosphate uptake. Phosphate uptake was blocked to about 25% of that of control at 100 ng/ml PMA. 3-Isobutyl-1-methyl-xanthine(IBMX) inhibited phosphate uptake. However, in the presence of IBMX, the inhibitory effect of exogenous cAMP was not significantly potentiated. Forskolin decreased phosphate transport. Acetylsalicylic acid did not inhibit phosphate uptake. The 1,2-dioctanoyl-sn-glycorol(DAG) and 1-oleoyl-2-acetyl-sn- glycerol(OAG) showed a inhibitory effect. However, staurosporine had no effect on phosphate uptake. When PMA and staurosporine were treated together, inhibition of phosphate uptake was not observed. In conclusion, phosphate uptake is stimulated by high sodium and low phosphate and pH 6.5 in the culture medium. Membrane potential and intracellular energy levels are also an important factor fer phosphate transport. Insulin and IGF-I stimulate phosphate uptake through a mechanisms that involve do novo protein and/or RNA synthesis and decrease of intracellular cAMP level. Also protein kinase C(PKC) is may play a regulatory role in transducing the insulin and IGF-I signal for phosphate transport in primary cultured proximal tubule cells.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.10
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• pp.1457-1465
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• 2012

Intestinal phosphate (Pi) absorption across the apical membrane of small intestinal epithelial cells is mainly mediated by the type IIb Na-coupled phosphate co-transporter (NaPi-IIb), but its expression and regulation in the chicken remain unclear. In the present study, we investigated the mRNA and protein levels of NaPi-IIb in three regions of chicken small intestine, and related their expression levels to the rate of net phosphate absorption. Our results showed that maximal phosphate absorption occurs in the jejunum, however the highest expression levels of NaPi-IIb mRNA and protein occurs in the duodenum. In response to a low-Pi diet (TP 0.2%), there is an adaptive response restricted to the duodenum, with increased brush border membrane (BBM) Na-Pi transport activity and NaPi-IIb protein and mRNA abundance. However, when switched from a low-(TP 0.2%) to a normal diet (TP 0.6%) for 4 h, there is an increase in BBM NaPi-IIb protein abundance in the jejunum, but no changes in BBM NaPi-IIb mRNA. Therefore, our study indicates that Na-Pi transport activity and NaPi-IIb protein expression are differentially regulated in the duodenum vs the jejunum in the chicken.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.5
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• pp.601-609
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• 1998

The present study was undertaken to examine the role of phospholipase $A_2\;(PLA_2)$ in oxidant-induced inhibition of phosphate transport in primary cultured rabbit renal proximal tubule cells. Uptakes of phosphate and glucose were dose-dependently inhibited by an oxidant t-butylhydroperoxide (tBHP), and the significant inhibition appeared at 0.025 mM of tBHP, whereas tBHP-induced alterations in lipid peroxidation and cell viability were seen at 0.5 mM. tBHP stimulated arachidonic acid (AA) release in a dose-dependent fashion. A $PLA_2$ inhibitor mepacrine prevented tBHP-induced AA release, but it did not alter the inhibition of phosphate uptake and the decrease in cell viability induced by tBHP. tBHP-induced inhibition of phosphate transport was not affected by a PKC inhibitor, staurosporine. tBHP at 0.1 mM did not produce the inhibition of $Na^+-K^+-ATPase$ activity in microsomal fraction, although it significantly inhibited at 1.0 mM. These results suggest that tBHP can inhibit phosphate uptake through a mechanism independent of $PLA_2$ activation, irreversible cell injury, and lipid peroxidation in primary cultured rabbit renal proximal tubular cells.

• The Korean Journal of Physiology and Pharmacology
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• v.11 no.2
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• pp.71-77
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• 2007

Cisplatin treatment increases the excretion of inorganic phosphate in vivo. However, the mechanism by which cisplatin reduces phosphate uptake through renal proximal tubular cells has not yet been elucidated. We examined the effect of cisplatin on $Na^+$-dependent phosphate uptake in opossum kidney (OK) cells, an established proximal tubular cell line. Cells were exposed to cisplatin for an appropriate time period and phosphate uptake was measured using $[^{32}P]$-phosphate. Changes in the number of phosphate transporter in membranes were evaluated by kinetic analysis, $[^{14}C]$phosphonoformic acid binding, and Western blot analysis. Cisplatin inhibited phosphate uptake in a time- and dose-dependent manner, and also the $Na^+$-dependent uptake without altering $Na^+$-independent uptake. The cisplatin inhibition was not affected by the hydrogen peroxide scavenger catalase, but completely prevented by the hydroxyl radical scavenger dimethylthiourea. Antioxidants were ineffective in preventing the cisplatin-induced inhibition of phosphate uptake. Kinetic analysis indicated that cisplatin decreased Vmax of $Na^+$-dependent phosphate uptake without any change in the Km value. $Na^+$-dependent phosphonoformic acid binding was decreased by cisplatin treatment. Western blot analysis showed that cisplatin caused degradation of $Na^+$-dependent phosphate transporter protein. Taken together, these data suggest that cisplatin inhibits phosphate transport in renal proximal tubular cells through the reduction in the number of functional phosphate transport units. Such effects of cisplatin are mediated by production of hydroxyl radicals.

• Korean Journal of Microbiology
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• v.33 no.1
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• pp.1-6
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• 1997

5-Enolpyruvylshikimate 3-phosphate(EPSP) synthase is the sixth enzyme of the shikimate pathway that synthesizes aromatic amino acids. The enzyme is a primary target for the glyphos'lte which is a broad-spectrum and environmetally safe herbicide. As a first step toward development of glyphpsate-resistant EPSP synthase, the EPSP synthase gene(aroA) was amplified by polymerase chain reaction and cluned into pET-25b vector. In this construct. designated pET-aro, the aroA gene is expressed under control of strong T7 promoter. and the EPSP synthase is produced as a fusion protein with pelB leader at N-terminus and HSV-tag and His-tag at C-terminus. When the pET-aro clone was induced to produce the enzyme, it was found that the EPSP synthase was successfully exported to peri plasmic space. The periplasmic transport was greatly dependent on the induction temperatures. Among the induction temperatures examined($25^{\circ}C$, $30^{\circ}C$, $34^{\circ}C$ and $37^{\circ}C$). induction at $34^{\circ}C$ gave rise to maximal periplasmic transport. The recomhinant EPSP synthase could have been purified hy $Ni^{2+}$ -affinity chromatography using the His-tag. and detected hy anti-HSV -tag antibody. The recombinant EPSP synthase also hound to phosphocellulose resin and was eluted hy shikimate 3-phosphate and phosphoenolpyruvate. as expected. The recombinant EPSP synthase purified from phosphocellulose resin showed typical EPSP synthase activity.