• The Korean Journal of Physiology
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• 제30권1호
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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.

• The Korean Journal of Physiology and Pharmacology
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• 제11권2호
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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.

• Applied Biological Chemistry
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• 제43권3호
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• pp.163-168
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• 2000

토양 및 수계에 집적, 유입되어 있는 과다한 인산의 제거에 이용 가능한 미생물 자원을 확보하기 위하여 활성오니에서 분리된 Acinetobacter lwoffi PO8의 배양조건 및 외부환경조건에 따른 인산흡수 양상을 조사하였다. 배양액 내 초기 pH가 $7.5{\sim}8.5$ 범위에서 균생육과 인산흡수율이 가장 높았으며, 탄소원으로 glycerol 및 arabinose을 첨가하였을 경우 각각 약 93 및 91%의 높은 인산 흡수율을 보였다. 질소원 형태에 따른 인산흡수양상은 아미노태 보다 암모늄염이 효과적이었으며, 특히, $NH_4NO_3$,와 $(NH_4)_2SO_4$가 각각 95와 96%의 인산흡수율을 나타냈다. 금속이온 중에서 $Co^{2+}$ 첨가 시 균생육이 제해되었으나, 이외의 다른 금속이온은 균의 성장 및 인산흡수에 큰영향을 미치지 않았다. 아미노산 중 arginine, methionine 및 lysine 등은 아미노산을 첨가하지 않은 경우보다 $10{\sim}20%$ 더 높은 인산흡수율을 나타냈었으며, 이때 배양기간 중 glucose의 공급으로 배지 중 잔존 인산이 완전히 제거되었다. 또한 $^{32}P$를 사용하여 균의 인산 분포를 조사한 결과로 세포내 흡수된 인산은 대부분 세포의 원형질 내에 polyphosphate의 형태로 분포되어 있었다.

• Journal of Acupuncture Research
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• 제17권3호
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• pp.208-218
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• 2000

This study was undertaken to determine if Salviae Radix (SR) exerts protective effect against oxidant-induced inhibition of phosphate uptake in renal proximal tubular cells. Membrane transport function and cell death were evaluated by measuring phosphate uptake and trypan blue exclusion, respectively, in opossum kidney (OK) cells, an established proximal tubular cell line. $H_2O_2$ was used as a model oxidant. $H_2O_2$ inhibited the phosphate uptake in a dose-dependent manner over the concentration range of 0.1-0.5 mM. Similar fashion was observed in cell death. However, the phosphate uptake was more vulnerable to $H_2O_2$ than cell death, suggesting that $H_2O_2$-induced inhibition of phosphate uptake is not totally attributed to cell death. Decreasedphosphate uptake was associated with ATP depletion and inhibition of $Na^+$-pump activity as determined by direct inhibition of $N^+-K^+$-ATPase activity. When cells were treated with $H_2O_2$ in the presence of 0.05% SR, the inhibition of phosphate uptake and cell death induced by $H_2O_2$ was significantly attenuated. SR restored ATP depletion and decreased $Na^+-K^+$-ATPase activity, and this is likely responsible for the protective effect of SR on decreased phosphate uptake. The protective effect of SR was similar to the $H_2O_2$ scavenger catalase. SR reacts directly with $H_2O_2$ to reduce the effective concentration of the oxidant. The iron chelator deferoxamine prevented the inhibition of phosphate uptake and cell death induced by $H_2O_2$, suggesting that $H_2O_2$-induced cell injury is resulted from an iron-dependent mechanism. These results indicate that SR exerts the protective effect against $H_2O_2$-induced inhibition of phosphate uptake by reacting directly with $H_2O_2$ like the $H_2O_2$scavenger enzyme catalase, in OK cells. However, the underlying mechanism remains to be explored.

• 한국수산과학회지
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• 제41권4호
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• pp.301-304
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• 2008

For bioremediation of the benthic layer uptake kinetics of phosphate by microphytobenthos Nitzschia sp.(JFH200406) were investigated. A short-term phosphate uptake revealed that the maximum uptake rate(${\rho}_{max}$) and half-saturation constant($K_s$) were 0.132 pmol/cell/hr and 502.6 ${\mu}M$, respectively. The maximum specific uptake rate calculated between ${\rho}_{max}$ and the phosphorus cell quota($Q_p$), calculated from Strathmann equation, was 14.4/day. The values of these parameters indicate that Nitzschia sp. accommodates well to surroundings of high phosphate, and can uptake over 14-times more than the phosphorus cell quota. Thus, microphytobenthos Nitzschia sp. may be a useful species for bioremediation of the benthic layer.

• 한국토양비료학회지
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• 제46권3호
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• pp.153-162
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• 2013

Most test methods for plant available soil phosphate are based on the extraction with a chemical solution. The objective of this study is to evaluate available phosphate of various tests at different soil phosphate levels. Two experiments were conducted as follows: i) Extracting capacities of soil phosphate tests - Mehlich III, Mehlich II, Bray I, Olsen, Kelowna, and Modified Lancaster(Mod. Lancaster) - were compared with that of Lancaster test for the soils collected from 32 paddy and 27 upland fields with various soil chemical properties. ii) Field trials on comparing to phosphate uptake by plant were accomplished by cultivating rice and corn plants in the pots filled with the soils. Available phosphate of Lancaster test was significantly correlated with those of Mehlich III, Mehlich II, Bray I, Olsen, Kelowna, and Mod. Lancaster. In upland soils, available phosphates of all the tests were curvilinearly regressed with phosphate uptake by corn. The determination coefficients ($R^2$) of the regression equation between available phosphate in soils and phosphate uptake by plants were ranged from 0.861 (Mehlich III) to 0.741 (Olsen). In paddy soils, the available phosphate measured by Mehlich III and Lancaster was significantly correlated with phosphate uptake by rice. In conclusion, Lancaster and Mehlich III tests could be used for predicting available phosphate in upland and paddy soils.

• 미생물학회지
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• 제29권2호
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• pp.117-122
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• 1991

In order to improve the removal ability of phosphate, Spheroplast fusions were performed among auxotrophic mutants of Aeromonas hydrophila isolated from waste water, named A13 and A14, Aci37 auxotrophic mutant of Acinetobactercalcoaceticus, and auxotrophic E. coli HR262/pCE27 carring pit gene. Eight fusants obtained from this experiment showed different biochemical characteristics. When the rate of phosphate uptake among fusants (F1-F8) was investigated in Phosphate Uptake Medium (PUM), F8 strain showed the highest rate for phosphate removal, 7 times as much as control after two hours incubation. The role of cations ($Mg^{++}$ ,$Ca^{++}$ , $K^{+}$ in phosphate uptade by F8 was also investigated in PUM without each salt. $K^{+}$ seemed to be crucial. Being compared with phosphate untake rate in PUM, that in PUM without $K^{+}$ was reduced 1.5 times. Therefore, by applying F8 strain and $K^{+}$ in practical environmental system, the increased efficiency in phosphate removal can be derived.

• 한국수산과학회지
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• 제40권2호
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• pp.95-101
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• 2007

We investigated the growth and phosphate uptake of a toxic dinoflagellate, Gymnodinium catenatum, isolated from Yeosuhae Bay, South Korea. A short-term phosphate uptake experiment revealed that its maximum uptake and the half-saturation constant were 1.39 pmol/cell/hr and $2.65{\mu}M$, respectively. In a semicontinuous culture, the maximum specific growth rate and minimum phosphorus cell quota of G. catenatum were 0.39/day and 1.27 pmol/cell, respectively. Thus, G. catenatum is a poor competitor in terms of inorganic nutrient use and is unlikely to form blooms in Yeosuhae Bay.

• 한국환경과학회지
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• 제14권9호
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• pp.873-879
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• 2005

This study was investigate to evaluate the phosphate uptake rate of green algae in relation to diurnal rhythm and algae control method. The phosphate uptake rates of Chlorella vulgaris and Ankistrodesmus convolutus increased in light period and decreased in dark period. On the contrary, those of Chlamydomonas sp. showed a peak in the late dark period. The differences among species in phosphate uptake in relation to diurnal rhythm were due to the severe competition among species and seemed to alleviate the competition for nutrient supplies. The compound of CellCaSi, Ca and Fe showed the effective removal of the phosphorus. The extracts from rice and barley straw exhibited a significant effect on the growth inhibition of Microcystis aeruginosa.

• The Korean Journal of Physiology and Pharmacology
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• 제13권1호
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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.