• BMB Reports
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• 제38권4호
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• pp.468-473
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• 2005

The energetics of nitrate uptake by intact cells of the halotolerant cyanobacterium Aphanothece halophytica were investigated. Nitrate uptake was inhibited by various protonophores suggesting the coupling of nitrate uptake to the proton motive force. An artificially-generated pH gradient across the membrane (${\Delta}pH$) caused an increase of nitrate uptake. In contrast, the suppression of ${\Delta}pH$ resulted in a decrease of nitrate uptake. The increase of external pH also resulted in an enhancement of nitrate uptake. The generation of the electrical potential across the membrane ($\Delta\psi$) resulted in no elevation of the rate of nitrate uptake. On the other hand, the valinomycin-mediated dissipation of $\Delta\psi$ caused no depression of the rate of nitrate uptake. Thus, it is unlikely that $\Delta\psi$ participated in the energization of the uptake of nitrate. However, $Na^+$-gradient across the membrane was suggested to play a role in nitrate uptake since monensin which collapses $Na^+$-gradient strongly inhibited nitrate uptake. Exogenously added glucose and lactate stimulated nitrate uptake in the starved cells. N, N'-dicyclohexylcarbodiimide, an inhibitor of ATPase, could also inhibit nitrate uptake suggesting that ATP hydrolysis was required for nitrate uptake. All these results indicate that nitrate uptake in A. halophytica is ATP-dependent, driven by ${\Delta}pH$ and $Na^+$-gradient.

• Journal of Pharmaceutical Investigation
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• 제25권2호
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• pp.137-143
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• 1995

The peptide transporter can be utilized for improving the bioavailability of compounds that are poorly absorbed. Characterization of the dipeptide uptake into the human intestinal epithelial cells, HT-29 was investigated. The uptake of tritiated glycylsarcosine $([^3H]-Gly-Sar,\;0.1\;{\mu}Ci/ml)$ was measured in confluent or subconfluent HT-29, Caco-2, and Cos-7 cells. Uptake medium was the Dulbecco's Modified Eagle's Media (DMEM) adjusted to pH 6.0. Both HT-29 and Caco-2 cells expressed the dipeptide transporter significantly (p<0.005) but Cos-7 did not. Certain portions of passive uptake were observed in all three cell lines. Uptake of Gly-Sar was largest at 7 days after plating HT-29 cells with significant inhibition with 25 mM cold Gly-Sar (p<0.05). but expression ratio of the dipeptide transporter was 0.7, suggesting lower expression. The effect of pH on Gly-Sar uptake was not significant in the range of pH 6 to 8. Gly-Sar uptake was also inhibited with 50 mM carnosine, 25 mM Gly-Sar, and 35 mM cephalexin significantly (p<0.05). From above results the dipeptide transporter was expressed well in HT-29 cells and was similar to that in the small intestine, suggesting that large amounts of mRNA of the transporter from the cells can be obtained.

• 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
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• 제22권2호
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• pp.281-293
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• 1988

가토 신장 근위세뇨관에서 분리한 brush border membrane vesicle(BBMV)과 basolateral membrane vesicle(BBMV)에서 rapid filtration 방법으로 PAH 이동에 대한 pH의 영향을 관찰하였다. BLMV에서 용액내 Na이 없을 때 외부 $pH(pH_0)$를 8.0에서 5.5까지 감소시켰을 때 probenecid-sensitive PAH 이동은 유의하게 증가되었다. 용액내 Na이 있을 때 $pH_0$가 8.0에서 6.0까지 변화하여도 PAH 이동에는 영향이 없었으나 5.5까지 더욱 감소시켰을 때 PAH 이동이 증가하였다. 그러나 vesicle 내 외에 pH gradient없이 $pH_0$를 내부 $pH(pH_1)$와 동일하게 변화시켰을 때 PAH 이동은 영향을 받지 않았다. PH gradient가 없을 때 시험된 pH범위에서 Na은 PAH 이동을 증가시켰다. BBMV에서도 BLMV에서와 유사한 pH 의존성을 보였으나 Na의 존재는 PAH 이동에 영향을 미치지 못하였다. BLMV에서 동력학적 분석 결과 일정한 $pH(pH_1)$에서 $pH(pH_0)$ 감소는 Km에 변화없이 PAH 이동에 대한 Vmax를 유의하게 증가시켰다. 이러한 결과로 BBMV에서 PAH 이동에 대한 pH의 영향은$OH^-/PAH$교환기전에 기인하는 것으로 추측되나 BLMV에서 pH 의존성은 음이온 교환기전만으로 설명될 수 없다. 또한 BLMV에서는 PAH 이동이 Na에 의존하나 BBMV에서는 Na에 의존하지 않음을 가르킨다.

• 분석과학
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• 제11권2호
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• pp.130-134
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• 1998

봄무우의 어린자엽을 이용하여 polyamine의 수송을 분석하였다. Spermidine(SPD)은 20분까지는 빠른 속도로 세포내로 수송되었고, 1시간 이후에는 포화상태에 도달하였다. SPD의 수송속도는 세포내에 polyamine(PA)의 농도가 증가될수록 감소되었다. PA의 수송속도는 외부배지의 pH에 의존되었고, 수송속도는 pH 7.5에서 최고에 도달되었다. $K_m-$과 $V_{max}-$값도 외부 배지의 pH에 의존되었다. KCl은 세포막전위를 낮추어 PA의 수송을 방해하였다.

• Environmental Engineering Research
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• 제10권5호
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• pp.257-263
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• 2005

A batch and a continuous type experiments were conducted to test the conditions for simultaneous phosphorus release and uptake, and denitrification, taking place in one process. The bacteria able to denitrify as well as to remove phosphorus were evaluated for the application to biological nutrient removal(BNR) process. In the batch-type experiment, simultaneous reactions of phosphorus release and uptake, and also denitrification were observed under anoxic condition with high organic and nitrate loading. However the rate and the degree of P release were lower than that occurred under anaerobic condition. BNR processes composed of anaerobic-anoxic-oxic(AXO), anoxic-anaerobic-oxic(XAO) and anoxic-oxic(XO) were operated in continuous condition. The anoxic reactors in each process received nitrate loading. In the AXO process, P release in anaerobic reactor and the luxury uptake in oxic reactor proceeded actively regardless to nitrate loading. However in XAO and XO processes, P release and luxury uptake occurred only with the nitrate loading less than $0.07\;kg{NO_3}^--N$/kgMLSS-d. With higher nitrate load, P release increased and the luxury uptake decreased. Therefore, it appeared that the application of denitrifying phosphorus-removing bacteria (DPB) to BNR process must first resolve the problem with decrease of luxury uptake of phosphorus in oxic reactor.

• Environmental Engineering Research
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• 제10권6호
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• pp.265-268
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• 2005

A batch-type study was conducted to investigate the phosphorus release and uptake under anaerobic and aerobic conditions according to organic loading changes. As organic loading increased, anaerobic P release increased but aerobic P uptake decreased. Where organic carbon contents remain high in aerobic conditions, PHB consumption within the microbial cells diminished, therefore it was found that in order to enhance P uptake rate, it should reach the endogenous growth stage where the entire organic loading was consumed.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.967-971
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• 2004

The kinetics of sulfite uptake were examined in a wild-type laboratory strain of Saccharomyces cerevisiae to determine if carrier-mediated sulfite uptake involved a proton symport, as previous studies on sulfite uptake have suggested both an active process and facilitated diffusion. Accumulation of intracellular sulfite was initially rapid and linear up to 50 sec. Uptake was saturable at final concentrations equal to or greater than 3 mM sulfite, and increased 2-fold in the presence of 2% glucose. Uptake was significantly reduced in cells pretreated with 100-500 $\mu$M carbonyl cyanide mchlorophenylhydrazone (CCCP) or 2,4-dinitrophenol (DNP), both of which dissipate proton gradients. Uptake was also significantly inhibited in the presence of 1 mM arsenate, an inhibitor of ATP synthesis. Extracellular alkalization was observed in cells incubated with 1-2 mM sulfite in a weak tartrate buffer at pH 3.5 and 4.5. These findings suggest that the bisulfite ion, $HSO_3^-$, an anionic form of sulfite, is taken up by a carrier-mediated proton symport. A met16 sull sul2 mutant, impaired in both sulfite formation and sulfate uptake, was found able to grow on a medium with sulfite as the sole Sulfur source, indicating that the sulfate transporters Sul1p and Sul2p are not required for sulfite uptake.

• The Korean Journal of Physiology and Pharmacology
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• 제3권5호
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• pp.529-538
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• 1999

This study was undertaken to examine the effect of ethanol on $Na^+ -dependent$ phosphate $(Na^+-P_i)$ uptake in opossum kidney (OK) cells, an established renal proximal tubular cell line. Ethanol inhibited ^Na^+-dependent$ component of phosphate uptake in a dose-dependent manner with $I_{50}$ of 8.4%, but it did not affect $Na^+-independent$ component. Similarly, ethanol inhibited $Na^+-dependent$ uptakes of glucose and amino acids (AIB, glycine, alanine, and leucine). Microsomal $Na^+-K^+-ATPase$ activity was not significantly altered when cells were treated with 8% ethanol. Kinetic analysis showed that ethanol increased $K_m$ without a change in $V_{max}$ of $Na^+-P_i$ uptake. Inhibitory effect of n-alcohols on $Na^+-P_i$ uptake was dependent on the length of the hydrocarbon chain, and it resulted from the binding of one molecule of alcohol, as indicated by the Hill coefficient (n) of 0.8-1.04. Catalase significantly prevented the inhibition, but superoxide dismutase and hydroxyl radical scavengers did not alter the ethanol effect. A potent antioxidant DPPD and iron chelators did not prevent the inhibition. Pyrazole, an inhibitor of alcohol dehydrogenase, did not attenuate ethanol-induced inhibition of $Na^+-P_i$ uptake, but it prevented ethanol-induced cell death. These results suggest that ethanol may inhibit $Na^+-P_i$ uptake through a direct action on the carrier protein, although the transport system is affected by alterations in the lipid environment of the membrane.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권4호
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• pp.221-224
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• 2002

Biomass of non-viable and dried brown marine algae Ecklonia radiata Turn. was used to examine Its cadmium uptake capability. Twelve different pretreatments on the algal biomass were prepared. Among these pretreatments, the algal biomass, which treated with 0.1 M NaOH and kept in water. bath (100$\^{C}$, 18 h) followed by washing with distilled water and squeezing, showed the highest amount of cadmium uptake as 1634 $\pm$ 195 mg/g dry biomass at pH 4.0 and 50$\^{C}$. Adsorption temperatures and pH levels played some Important role In cadmium uptake. However, cadmium uptake decreased dramatically at a lower pH than 4.0. Freundlich adsorption isotherm showed potent cadmium uptake capacity of the non-viable biomass. Pretreatments on the non-viable algal biomass shown in this study nay enhance the Eadmium removal in the industrial wastewater.