• 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.

• Journal of Life Science
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• v.19 no.5
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• pp.566-572
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• 2009

A recombinant plasmid, pDH3, was obtained from the genomic library of Serattia marcescens KCTC 2172, and several recombinant subclones constructed from pDH3. The nucleotide sequence of a 5,137 bp segment, pPH4, was determined and three open reading frames were detected. The three ORFs encoded the phosphate specific transport (pst) operon, which was pstC, pstA, and pstB, with the same direction of transcription. Comparison of the pst operon of S. marcescens with that of other organisms revealed that the genes for pstS and phoU were missing. A potential CRP bonding site and pho box sequence was found in the upstream of the putative promoter at the regulatory region. Analysis of the nucleotide sequence showed that homology in amino acid sequences between the PstC protein and Yersinia sp., Vibrio sp., and Pseudomonas sp. were 49, 37 and 33%, respectively. The PstA protein and Yersinia sp., Vibrio sp., and Pseudomonas sp. showed homologies of 64, 51, and 47%, respectively. PstB protein and Methanocaldococcus sp., E. coli, and Mycoplasma sp. showed homologies of 60, 50, and 48%, respectively. The pst genes could be expressed in vivo and positively regulated by cAMP-CRP. The E. coli strain harboring plasmid pPH7, with pst genes, increased with the transport of phosphate.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.12
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• pp.25-29
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• 1972

This experiment was carried out to investigate the role of Silicon accumlated in rice plant under different conditions of light and humidity, using radioisotopes Ca-45, Mn-54, and P-32. This results obtained in are as follows; 1. Light effect is more severe in phosphate uptake by rice plant than is calcium. Amounts of phosphate uptake in light condition is six times more than in dark conditions, while that of calcium is double. 2. Change of relative humidity affects calcium absorption and transport from root to shoot. It seems not to be influenced in phosphate and manganese uptake by relative humidity. 3. More uptake of each element Ca-45, P-32, or Mn-54 was found in the rice plant applied with silicic acid. It is considered that there must be some relationship between silicon content and ion uptake in rice plant. 4. The transport ratio of nutrient from root to shoot shows a specific pattern that calcium is approximately 1.0 manganese 0.5 and phosphate 0.2 respectively.

• Proceedings of the Korean Society of Life Science Conference
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• 2002.03a
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• pp.99-99
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• 2002

• Clinical and Experimental Pediatrics
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• v.50 no.3
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• pp.230-235
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• 2007

In the early neonatal period, the neonate is challenged by the loss of the placental calcium transport and manifests a quick transition, from an environment in which PTHrP plays an important role to a PTH- and 1,25-dihydroxyvitamin D-controlled neonatal milieu. Disturbances in mineral homeostasis are common in the neonatal period, especially in premature infants and infants who are hospitalized in an intensive care unit. In many cases these disturbances are thought to be exaggerated responses to the normal physiological transition from the intrauterine environment to neonatal independence. Some disturbances in calcium and phosphate homeostasis are the result of genetic defects, which in many instances can now be identified at the molecular level. Although fetus develop remarkably normally in the presence of maternal calcium, PTH and vitamin D deficiency, the neonates demonstrate abnormalities that are consequences of the prior abnormal maternal calcium homeostasis. Evaluation and management of hypocalcemia and hypercalcemia in neonate requires specific knowledge of perinatal mineral physiology and the unique clinical and biochemical features of newborn mineral metabolism.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.11
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• pp.1673-1679
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• 2008

In order to study the biological function of gapdh gene in yak, and prove whether the gapdh gene was a useful intra-reference gene that can be given an important role in molecular biology research of yak, the cDNA sequence encoding glyceraldehyde-3-phosphate dehydrogenase from yak was cloned by the RT-PCR method using gene specific PCR primers. The sequence results indicated that the cloned cDNA fragment (1,008 bp) contained a 1,002 bp open reading frame, encoding 333 amino acids (AAs) with a molecular mass of 35.753 kDa. The deduced amino acids sequence showed a high level of sequence identity to Bos Taurus (99.70%), Xenopus laevis (94.29%), Homo sapiens (97.01%), Mus musculus (97.90%) and Sus scrofa (98.20%). The expression of yak's gapdh gene in heart, spleen, kidney and brain tissues was also detected; the results showed that the gapdh gene was expressed in all these tissues. Further analysis of yak GAPDH amino acid sequence implied that it contained a complete glyceraldehyde-3-phosphate dehydrogenase active site (ASCTTNCL) which ranged from 148 to 155 amino acid residues. It also contained two conserved domains, a NAD binding domain in its N-terminal and a complete catalytic domain of sugar transport in its C-terminal. The phylogenetic analysis showed that yak and Bos taurus were the closest species. The prediction of secondary structures indicated that GAPDH of yak had a similar secondary structure to other isolated GAPDH. The results of this study suggested that the gapdh gene of yak was similar to other species and could be used as the intra-reference to analyze the expression of other genes in yak.

• Journal of Applied Biological Chemistry
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• v.50 no.2
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• pp.58-62
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• 2007

Identification of Dual-specificity protein phosphatase (DSP) substrates is essential in revealing physiological roles of DSPs. We isolated VHZ-interacting proteins from extracts of 293T cells overexpressing a VHZ (C95S, D65A) mutant known to be substrate- trapping mutant. Analysis of specific proteins bound to VHZ by 2D gel electrophoresis and mass spectroscopy revealed that these proteins contained Chaperonin containing TCP1, Type II phosphatidylinositol phosphate kinase ${\gamma}$, Intraflagellar transport 80 homolog, and Kinesin superfamily protein 1B. VHZ-interacting proteins showed that VHZ is involved in many important cellular signal pathways such as protein folding, molecular transportation, and tumor suppression.

• Molecules and Cells
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• v.28 no.5
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• pp.479-487
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• 2009

Glutathione reductase (GR) is an enzyme that recycles a key cellular antioxidant molecule glutathione (GSH) from its oxidized form (GSSG) thus maintaining cellular redox homeostasis. A recombinant plasmid to overexpress a GR of Brassica rapa subsp. pekinensis (BrGR) in E. coli BL21 (DE3) was constructed using an expression vector pKM260. Expression of the introduced gene was confirmed by semi-quantitative RT-PCR, immunoblotting and enzyme assays. Purification of the BrGR protein was performed by IMAC method and indicated that the BrGR was a dimmer. The BrGR required NADPH as a cofactor and specific activity was approximately 458 U. The BrGR-expressing E. coli cells showed increased GR activity and tolerance to $H_2O_2$, menadione, and heavy metal ($CdCl_2$, $ZnCl_2$ and $AlCl_2$)-mediated growth inhibition. The ectopic expression of BrGR provoked the co-regulation of a variety of antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase, and glucose-6-phosphate dehydrogenase. Consequently, the transformed cells showed decreased hydroperoxide levels when exposed to stressful conditions. A proteomic analysis demonstrated the higher level of induction of proteins involved in glycolysis, detoxification/oxidative stress response, protein folding, transport/binding proteins, cell envelope/porins, and protein translation and modification when exposed to $H_2O_2$ stress. Taken together, these results indicate that the plant GR protein is functional in a cooperative way in the E. coli system to protect cells against oxidative stress.

• Korean Journal of Environmental Agriculture
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• v.42 no.1
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• pp.52-62
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• 2023

This study focused on determining the specific causes and prevention methods of mass fish deaths occurred in five reservoirs (Gagugi, Neupgokgi, Danggokgi, Sagokji, and Hangokji) in Andong-si. For this purpose, a survey of agricultural land and livestock in the upper part of the reservoirs and analysis of water quality in the reservoir irrespective of whether it rains or not were conducted. We attempted to examine the changes in dissolved oxygen (DO) in the surface and bottom layers of reservoirs and changes in DO depending on the amount of livestock compost and time. Based on the above investigations, treatment plans were established to efficiently control the inflow of contaminated water into reservoirs. The rainfall and farmland areas in the upper part of the reservoir were investigated using Google and aviation data provided by the Ministry of Land, Infrastructure, and Transport. The current status of livestock farms distributed around the reservoirs was also examined because compost from these farms can flow into the reservoir when it rains. Various water quality parameters, such as phosphate phosphorus (PO₄-P) and ammonium nitrogen (NH₃-N), were analyzed and compared for each reservoir during the rainy season. Changes in the DO concentration and electrical conductivity (EC) were also observed at the inlet of the reservoir during raining using an automated instrument. In addition, DO was measured until the concentration reached 0 ppm in 10 min by adding livestock compost at various concentrations (0.05%, 0.1%, 0.3%, and 0.5% by wt.), where the concentration of the livestock compost represents the relative weight of rainwater. The DO concentration in the surface layer of reservoirs was 3.7 to 5.3 ppm, which is sufficient for fish survival. However, the fish could not survive at the bottom layer with DO concentration of 0.0-2.1 ppm. When the livestock compost was 0.3%, DO required 10-19 h to reach 0 ppm. Considering these results, it was confirmed that the DO in the bottom layer of the reservoir could easily change to an anaerobic state within 24 h when the livestock compost in the rainwater exceeds 0.3%. The results show that the direct cause of fish mortality is the inflow of excessive livestock compost into reservoirs during the first rainfall in spring. All the surveyed reservoirs had relatively good topographical features for the inflow of compost generated from livestock farms. This keeps the bottom layer of the reservoir free of oxygen. Therefore, to prevent fish death due to insufficient DO in the reservoir, measures should be undertaken to limit the amount of livestock compost flowing into the reservoir within 0.3%, which has been experimentally determined. As a basic countermeasure, minerals such as limestone, dolomite, and magnesia containing calcium and magnesium should be added to the compost of livestock farms around the reservoir. These minerals have excellent pollutant removal capabilities when sprayed onto the compost. In addition, measures should be taken to prevent fish death according to the characteristics of each reservoir.