• Title/Summary/Keyword: phosphate depletion

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Alterations in Membrane Transport Function and Cell Viability Induced by ATP Depletion in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Lee, Sung-Ju;Kwon, Chae-Hwa;Kim, Yong-Keun
    • 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.

Effects of Phosphate, Precursor and Exogenous Berberine on the Production of Alkaloid in Plant Cell Cultures

  • Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.79-83
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    • 1991
  • The effects of phosphate concentration in the medium, feeding of biosynthetic precursor, and the addition of exogenous berberine on cell growth and berberine production were studied in cell suspension cultures of Thalictrum rugosum. The depletion of phosphate in the medium enhanced the specific productivity up to twofold with significant release of berberine into the medium. Extracellular berberine was 19% of the total in the culture without phosphate while it was 2-5% of total berberine in the culture with even low amounts of phosphate. Precursor feeding was not effective in enhancing alkaloid formation. Initial presence of exogenous berberine did not have much effect on cell growth and alkaloid production. It was found that the cells have the capacity to take up large quantities of berberine. When $500{\;}mg{\cdot}l^{-1}$ of berberine was added exogenously at the beginning, 81% of total berberine was found in the cells.

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Establishment of Phosphorus Flow Model in Urban Area using Material Flow Analysis

  • Lee, Mina;Kim, Kye-Hoon
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.2
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    • pp.80-84
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    • 2014
  • Phosphorus (P) is an essential nutrient for all living organisms. P is mostly obtained from mined rock phosphate. However, existing rock phosphate reserves could be exhausted in the next 50-100 years. As Korea is totally dependent on imported rock phosphate, we should seek for solution to overcome the P depletion by efficient use and recycling. For this, this study suggested a P flow model to identify the location and flow route of P in urban area based on traditional material flow analysis. The type of P entering the urban areas are fertilizer, food and feed. Each type of P is used in agriculture, human consumption and animal husbandry. After going through each process, P is moved to waste management facilities within food waste, excreta and sewage. Some portion of P in waste are buried, incinerated and discharged, which can be reservoir of P in the future.

The Effect of Salviae Radix on Oxidat-Inhibition of Phosphate Uptake in Renal Proximal Tubular Cells (단삼약침액(丹蔘藥鍼液)이 신장(腎臟) 근위세뇨관세포(近位細尿管細胞)에서 산화제(酸化劑)에 의한 인산(燐酸)의 이동억제(移動抑制)에 미치는 영향(影響))

  • Lee, Ho-Dong;Youn, Hyoun-Min;Jang, Kyung-Jeon;Song, Choon-Ho;Ahn, Chang-Beohm
    • Journal of Acupuncture Research
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    • v.17 no.3
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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.

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Espression of Alkaline Phosphatase Gene from Kluyveromyces fragilis in E. coli and S. cerevisiae (Kluyveromyces fragilis의 Alkaline Phosphatase 유전자의 E. coli 및 S. cerevisiae 에서의 발현)

  • Park, Soo-Young;Hwang, Seon-Kap;Lee, Dong-Sun;Kim, Jong-Guk;Nam, Joo-Hyun;Hong, Soon-Duck
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.131-137
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    • 1995
  • The alkaline phosphatase (K-ALPase) gene of Kluyveromyces fragilis has been cloned (1) and determined its base sequences (2) previously in our laboratory. When the K-ALPase gene was expressed in Escherichia coli and Saccharomyces cerevisiae, it showed a constitutive activity in E. coli, and a derepressed activity in S. cerevisiae in phosphate-limited medium. Northem hybridization experiment was performed to elucidate the transcription level of the K-ALPase gene. Northern experiment showed that transcription level of K-ALPase gene in S. cerevisiae was higher in phosphate depletion, but it was higher in high phosphate medium than in phosphate limited medium in K. fragilis. The transcription initiation site of the K-ALPase gene was determined by primer extension analysis. It matched nucleotide position - 169 in relation to the putative trnslational start site.

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Influence of Carbon and Nitrogen Sources in Solubilization of Hardly Soluble Mineral Phosphates by Penicillium Oxalicum CBPS-Tsa

  • Kim, Eun-Hee;Sundaram, Seshadri;Park, Myoung-Su;Shin, Wan-Sik;Sa, Tong-Min
    • Korean Journal of Environmental Agriculture
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    • v.22 no.3
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    • pp.197-202
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    • 2003
  • Phosphorus is one of the major plant growth limiting nutrients, despite being abundant in soils in both inorganic and organic forms. Phosphobioinoculants in the form of microorganisms can help in increasing the availability of accumulated phosphates for plant growth by solubilization. Penicillium oxalicum CBPSTsa, isolated from paddy rhizosphere, was studied for its phosphate solubilization. The influence of various carbon sources like glucose, sucrose, mannitol and sorbitol and nitrogen sources like arginine, sodium nitrate, potassium nitrate, ammonium chloride and ammonium sulphate were evaluated using liquid media with tricalcium phosphate (Ca-P), ferric phosphate (Fe-P) and aluminium phosphate (Al-P). Maximum soluble phosphate of 824 mg/L was found in the amendment of sucrose-sodium nitrate from 5 g/L of Ca-P. Mannitol, sorbitol, and ariginine were poor in phosphate solubilization. While sucrose was better carbon source in solubilization of Ca-P and Al-P, glucose fared better in solubilization of Fe-P. Though all the nitrogen sources enhanced P solubilization, nitrates were better than ammonium In the amendments of ammonium chloride and ammonium sulphate, higher uptake of available phosphates by the fungus was found, and this resulted in depletion of available P in Fe-P amendment Phosphate solubilization was accompanied by acidification of the media, and the highest pH decrease was observed in glucose amendment Among the nitrogen sources, ammonium chloride favored greater pH decrease.

Dynamic Gene Expression Profiling of Escherichia coli in Carbon Source Transition from Glucose to Acetate

  • Oh Min-Kyu;Cha Mee-Jeong;Lee Sun-Gu;Rohlin Lars;Liao James C.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.4
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    • pp.543-549
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    • 2006
  • DNA microarray was used to study the transcription profiling of Escherichia coli adapting to acetate as a sole carbon source. Bacteria grown in glucose minimal media were used as a reference. The dynamic expression levels of 3,497 genes were monitored at seven time points during this adaptation. Among the central metabolic genes, the glycolytic and glucose phosphotransferase genes were repressed as the bacteria entered stationary phase, whereas the glyoxylate pathway, TCA cycle, and gluconeogenic genes were induced. Distinct induction or repression patterns were recognized among different pathway genes. For example, the repression of glycolytic genes and the induction of gluconeogenic ones started immediately after glucose was depleted. On the other hand, the regulation of the pentose phosphate pathway genes and glyoxylate genes gradually responded to the glucose depletion or was more related to growth in acetate. When the whole genome was considered, many of the CRP, FadR, and Cra regulons were immediately responsive to the glucose depletion, whereas the $\sigma^s$, Lrp, and IHF regulons were gradually responsive to the glucose depletion. The expression profiling also provided differential regulations between isoenzymes; for example, malic enzymes A (sfcA) and B (maeB). The expression profiles of three genes were confirmed with RT-PCR.

Effect of Scutellaria Baicalensis Georgi Extraction (SbGE) on H2O2-induced Inhibition of Phosphate Transport in Renal Epithelial Cells (황금약침액(黃芩藥鍼液)이 신장상피세포(腎臟上皮細胞)에서의 H2O2에 의한 인산염(燐酸鹽) 운반(運搬)의 억제(抑制)에 미치는 영향(影響))

  • Cho, Eun-jin;Youn, Hyoun-min;Jang, Kyung-jeon;Song, Choon-bo;Ahn, Chang-beobm
    • 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.

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Continuous removal of phosphorus in water by physicochemical method using zero valent iron packed column (영가철 충진 컬럼을 이용한 연속적인 물리화학적 수중 인 제거)

  • Jeong, Jooyoung;Ahn, Byungmin;Kim, Jeongjoo;Park, Jooyang
    • Journal of Korean Society of Water and Wastewater
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    • v.27 no.4
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    • pp.439-444
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    • 2013
  • Excessive phosphorus in aquatic systems causes algal bloom resulting in eutrophication, DO depletion, decline in recreational value of water and foul tastes. To treat wastewater containing phosphorus including effluent of wastewater treatment plant, the continuous experiments were performed by using electrochemical way. The spherical ZVI and silica sand which act as physical filter are packed at appropriate volume ratio of 1:2. Electric potential is applied externally which can be changed as per the operational requirement. The results indicate that optimum hydraulic retention time of 36 minutes (10 mL/min at 1 L reactor) was required to meet the effluent standards. Lower concentrations of phosphorus (<10 mg/L as phosphate) were removed by precipitation by contact with iron. Thus, additional electric potential was not required. In order to remove high concentration phosphorus around 150 mg/L as phosphate, external electric potential of 600 V was applied to the reactor.

A Study On Structure Change of Binding Actin and Moysin On The Contracting Muscle Membrane (수축중인 근육막에서 actin과 myosin 결합의 구조변화에 관한 연구)

  • Kim, Duck-Sool;Park, Keun-Ho
    • Journal of the Korean Applied Science and Technology
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    • v.25 no.3
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    • pp.380-387
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    • 2008
  • The effects of the applied stretch and MgADP binding on the structure of the actin and myosin cross-bridges in rabbit fibers in the rigor state have been investigatedwith improved resolution by x-ray diffraction using synchrotron radiation. To clarify the structure of the ATP hydrolysis intermediates formed by actin and myosin cross-bridges,the effects of various phosphate analogs in the of MgADP on the structure of the thin and thick filaments in glycerinated rabbit muscle fibers in the rigor state investigated by x-ray diffraction with a short exposure time using synchrotron radiation. These results strongly suggest that when MgADP and phosphate analogs such as metallofluorides(BeF3 and AlF4)and vanadate(VO4(Vi)) were added the rigor fibers in the presence of the ATP-depletion backup system, the intensities of the actin-based layer lines were markedly weakened. We found that the intensity of the 14.5 nm-based meridional reflections increase by 20-50% when phosphate analogs such as metallofluorides(BeF3 and AlF4) and vanadate(VO4(Vi)) was added to the rigor muscle.