• Title/Summary/Keyword: H-uptake

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Effects of $aroP^{-}$ mutation on the tryptophan excretion in escherichia coli ($aroP^{-}$변이가 E.coli에서 트립토판 방출에 미치는 영향)

  • 지연태;안병우;이세영
    • Korean Journal of Microbiology
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    • v.23 no.1
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    • pp.9-12
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    • 1985
  • As a part of the host cell development for a amplified recombinant trp operon, $aroP^-$ mutation was introduced in a E. coli host strain. $aroP^-$ mutation was induced by transposon Tn10 and transduced into the E. coli host cell by bacteriophage P1Kc. The effect of $aroP^-$ mutation on the excretion of tryptophan in E. coli $trpR^{-ts}/ColE_1 -trp^+$ cells was investigated. Mutant lacking the general aromatic transport system was resistant to ${\beta}-2-thienylalanine\;(2{\times}10^{-4}\;M)$, p-fluorophenylalanine $(2{\times}10^{-4}M)$, or 5-methyltryptophan $(2{\times}10^{-4}\;M.)[^3H]-tryptophan$ uptake of the $aroP^-$ mutant strain was reduced considerably as compared with $aroP^+$ counterpart. The rate of $[^3H]-tryptophan$ uptake of the $aroP^-$ mutant strain treated with $NaN_3(3{\times}10^{-2}\;M)$ was much less affected than that of $aroP^+$ counterpart. The $aroP^-$ transductants increased the tryptophan excretion from E. coli $trpR^{-ts}/ColE_1 -trp^+$ four times more than $aroP^+$ counterpart.

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Experimental Design in Laboratory for Ecological Restoration in the Slag Dumping Area

  • Kim, Jin-Man;Kwak, Seck-Nam
    • Journal of Environmental Science International
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    • v.18 no.10
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    • pp.1065-1070
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    • 2009
  • Experimental design for ecological restoration approach was investigated from the P' company's slag dumping area with higher pH value. The degree of pH recovery was measured by the injection of carbon dioxide from the LNG exhaust gas, and the residual carbon, for example, calcium and carbonate which can be controlled by artificial seaweed beds. The degree of adaptability from 3 algaes (Undaria pinnatifida, Sargassum horneri and Ecklonia stolonifera) and uptake nutrient function of Ecklonia stolonifera chosen in the first treatment were measured in the laboratory to determine the transplanting algae in artificial seaweed beds. The higher value of pH was decreased to 7.0~8.5 by injection of LNG exhaust gas with flow rate $20\;m^3/min$. In the experiment design at laboratory, the upper part of frond of Undaria pinnatifida and Sargassum horneri began to decay, and the color changed after 10 days. However, those of Ecklonia stolonifera were after 14~20 days. The uptake rate of nutrient from Ecklonia stolonifera was higher than those of other algaes, and those was similar pattern in the control (e.g. seawater); The DIN concentration uptake of Ecklonia stolonifera was $1.88\;{\mu}g/L/hr$ avg. in leachate, but $2.19\;{\mu}g/L/hr$ avg. in seawater. However, the ${PO_4}^3$-P concentration uptake was $0.18\;{\mu}g/L/hr$ avg. in leachate, but $0.31\;{\mu}g/L/hr$ avg. in seawater. These result indicated Ecklonia stolonifera uptaked these nutrients in the leachate as well as in seawater, and it may suggested for this species to transplant for constructing artificial seaweed beds.

Regulation of Choline Transport by Oxidative Stress at the Blood-Brain Barrier In Vitro Model

  • Kang, Young-Sook;Lee, Hyun-Ae;Lee, Na-Young
    • Biomolecules & Therapeutics
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    • v.16 no.1
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    • pp.14-20
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    • 2008
  • In the present study, we examined how the transport of choline is regulated at the blood-brain barrier (BBB) under the central nervous system (CNS) cellular damages by oxidative stress using a conditionally immortalized rat brain capillary endothelial cells (TR-BBB), in vitro the BBB model. It was also tested whether the choline uptake is influenced by membrane potential, extracellular pH, protonophore (FCCP) and amiloride in TR-BBB cells. In result, $[^3H]choline$ uptake was inhibited by FCCP and dependent on extracellular pH. The treatment of TR-BBB cells with 20 ng/mL tumor necrosis $factor-{\alpha}$ $(TNF-{\alpha})$, 10 ng/mL lipopolysaccharide (LPS), 100 ${\mu}M$ diethyl maleate (DEM) and 100 ${\mu}M$ glutamate resulted in 3.0-fold, 2.6-fold, 1.8-fold and 2.0-fold increases of $[^3H]choline$ uptake at the respective peak time, respectively. In contrast, hydrogen peroxide and raffinose did not show any significant effects on choline uptake. In addition, choline efflux was significantly inhibited by $TNF-{\alpha}$, LPS and DEM producing cell damage states. In conclusion, the influx and efflux transport system for choline existed in TR-BBB cell line and this process was affected by several oxidative stress inducing agents.

New CPS-PPEES blend membranes for CaCl2 and NaCl rejection

  • Chitrakar, Hegde;Arun, M. Isloor;Mahesh, Padaki;Ahmad, Fauzi Ismail;Lau, W.J.
    • Membrane and Water Treatment
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    • v.3 no.1
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    • pp.25-34
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    • 2012
  • Carboxylated polysulfone (CPS), poly (1,4-phenylene ether ethersulfone) (PPEES), membranes were prepared and used for the separation of NaCl and $CaCl_2$, in efficient way with less energy consumption. In this work, nanofiltration and reverse osmosis membranes were employed to the salt rejection behavior of the different salt solutions. The influence of applied pressure (1-12 bar), on the membrane performance was assessed. In CM series of membranes, $CM_1$ showed maximum of 97% water uptake and 36% water swelling, whereas, $CM_4$ showed 75% water uptake and 28% water swelling. In RCM series, $RCM_1$ showed 85% water uptake and 32% water swelling whereas, in $RCM_4$ it was 68% for water uptake and 20% for water swelling. Conclusively reverse osmosis membranes gave better rejection whereas nanofiltration membrane showed enhanced flux. CM1 showed 58% of rejection with 12 L/($m^2$ h) flux and $RCM_1$ showed 55% of rejection with 15 L/($m^2$ h) flux for 0.1 wt.% NaCl solution. Whereas, in 0.1 wt.% $CaCl_2$ solution, membrane $CM_1$ showed 78% of rejection with 12 L/($m^2$ h) flux and $RCM_1$ showed 63% rejection with flux of 9 L/($m^2$ h).

Functional Characterization and Regional Expression of Dopamine Transporter (도파민 수송체의 기능적 특성 및 발현에 관한 연구)

  • 이상훈;이송득;성기욱;이동섭;이용성;고재경
    • YAKHAK HOEJI
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    • v.39 no.2
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    • pp.161-168
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    • 1995
  • Brain dopamine systems play a central role in the control of movement, hormone release, and many complex behavior. The action of dopamine at its synapse is terminated predominately by high affinity reuptake into presynaptic terminals by dopamine transporter (DAT). The dopamine transporter(DAT) is membrane protein localized to dopamine-containing nerve terminals and closely related with cocaine abuse, Parkinsonism, and schizophrenia. In present study, the recombinant plasmid pRc/CMV-DAT, constructed by subcloning of a cDNA encoding a bovine DAT into eukaryotic expression vector pRc/CMV, was stably transfected into CV-1 cells(monkey kidney cell line). The DAT activities in the cell lines selected by Geneticin$^{R}$ were determined by measuring the uptake of $[^3H]$-dopamine. The transfected cell lines showed 30-50 fold higher activities than untransfected CV-1 cell line, and this result implies that DAT is well expressed and localized in transfected cells. The transfected cells accumulated $[^3H]$-dopamine in a dose-dependent manner with a $K_{m}$ of 991.6nM. Even though high doses of norepinephrine, epinephrine, serotonin, and choline neurotransmitters inhibited the uptake of $[^3H]$-dopamine, DAT in transfected cell line was proven to be much more specific to dopamine. The psychotropic drugs such as GBR12909, CFT, normifensine, clomipramine, desipramine, and imipramine inhibited significantly the dopamine uptake in tissue culture cells stably transfected with DAT cDNA. Radioactive in situ hybridization was done to map the cellular localization of DAT mRNA-containing cells in the adult rat central nervous system. The strong hybridization signals were detected only in the substantia nigra pars compacta and ventral tegmental area. The restricted anatomical localization of DAT mRNA-containing cells confirms the DAT as a presynaptic marker of dopamine-containing cells in the rat brain.

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Characterization of Putrescine Uptake in Hamster Amelanocytic Melanoma AMEL-3 Cells

  • Garcia-Fernandez, Antonio J.;Rodriguez, Rosa A.;Perez-Pertejo, Yolanda;Balana-Fouce, Rafael
    • Molecules and Cells
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    • v.20 no.1
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    • pp.127-135
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    • 2005
  • The uptake of putrescine, spermidine and spermine by Fortner's hamster amelanocytic melanoma AMEL-3 cells was observed in this study to be time-dependent, temperature-sensitive, pH-dependent and saturable. Metabolic poisons nullified polyamine uptake, an indication that this is an energy-requiring mechanism. The presence of $Na^+$ ions was found to be requisite to full activity. Valinomycin, gramicidin, monensin and the calcium ionophore calcimycin were also observed to inhibit the process substantially. The transporter active site would seem to contain sulfhydryl groups. Other diamines and polyamine analogues, as well as cationic diamidines, suppressed putrescine uptake. The presence of the ornithine decarboxylase inhibitor DFMO in the culture medium induced putrescine inflows. Putrescine, in turn, induced the negative expression of the carrier, thus suggesting that this influx mechanism is governed by up/down regulation. The cationic diamidine CGP 40215A and its analogue CGP039937A competitively inhibited putrescine transport, with Ki values of 1.9 and $15{\mu}M$, respectively. The role of polyamine uptake in these cultures is discussed.

CYTOTOXICITY OF D-GALACTOSAMINE ON PRIMARY CULTURES OF ADULT RAT HEPATOCYTES

  • Yang, K.H.;Park, Kwan-Ha;Kim, Byung-Sam
    • Toxicological Research
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    • v.3 no.2
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    • pp.73-80
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    • 1987
  • Primary cultures of adult rat hepatocytes were used to study the cytotoxicity of D-galactosamine. Hepatocytes were isolated by a collagenase perfusion technique and maintained as monolayers in serum-free medium on collagen-coated culture dishes. Treatment of galactosamine to the culture markedly inhibited the uptake of ${\alpha}$-aminoisobutyric acid (AIB) inducible with glucagon and dexamethasone. At0.1 mM of galactosamine, AIB uptake was inhibited significantly when treated for 12 hr. At higher doses (0.25, 0.5 and 1.0mM), a significant inhibition was noticed after 1 hr exposure. Generally the magnitude of the inhibition was related to the dose and treatment time of galactosamine. Treatment of galactosamine also produced a dose- and treatment time-related suppression of the tyrosine aminotransferase (TAT) induction caused by dexamethasone. Meanwhile, uptake of ouabain was not affected by the treatment of galactosamine. The viability of the hepatocytes was decreased only slightly by the treatment of galactosamine; more than 87% of the cells excluded tryphane blue when treated 1 mM galactosamine for 12 hr. Galactosamine induced depressions of AIB uptake and TAT activity were prevented by the simultaneous addition of uridine to the culture. D-Galactosamine, cytotoxicity, hepatocytes culture, ${\alpha}$-aminoisobutyric acid uptake, tyrosine aminotransferase.

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Effects of Long-Term Fertilization for Cassava Production on Soil Nutrient Availability as Measured by Ion Exchange Membrane Probe and by Corn and Canola Nutrient Uptake

  • Hung T. Nguyen;Anh T. Nguyen;Lee, B.W.;J. Schoenau
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.2
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    • pp.108-115
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    • 2002
  • The effects of long-term fertilization on soil properties and nutrient availability are not well documented for cassava cultivation in Vietnam. In 1990, a field research plots were established with 12 treatments to test the effect of different rates of nitrogen (N), phosphorus (P) and potassium (K) on soil properties in Acrisols at Thai Nguyen University in Northern Vietnam. In 1999, composite soil samples (0 to 20cm depth) were collected from eight selected plots for measurements of nutrient supply rates by ion exchange membrane probes and for growing corn and canola in a growth chamber with and without added lime. Generally, long-term nitrogen (N) fertilization increased available N supply rates but decreased available potassium (K) and magnesium (Mg). Long-term phosphorus(P) applications increased canola N, calcium (Ca) and Mg uptake. Canola P uptake increased with increased P rates only when lime was added. Long-term K applications increased canola N, K, Ca, Mg uptake but only significantly increased corn N uptake. Liming significantly increased uptake of N, P, K, Ca, Mg and S for both corn and canola. However, N $H_{4-}$N, K and Mg soil supply rates were reduced when lime was added, due to competition between Ca from the added lime and other nutrients.

Effects of insulin and IGF on growth and functional differentiation in primary cultured rabbit kidney proximal tubule cells - Effects of IGF-I on Na+ uptake - (초대배양된 토끼 신장 근위세뇨관세포의 성장과 기능분화에 대한 insulin과 IGF의 효과 - Na+ uptake에 대한 IGF-I의 효과 -)

  • Han, Ho-jae;Park, Kwon-moo;Lee, Jang-hern;Yang, IL-suk
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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    • pp.783-794
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    • 1996
  • It has been suggested that ion transport systems are intimately involved in mediating the effects of growth regulatory factors on the growth of a number of different types of animal cells in vivo. The functional importance of the apical membrane $Na^+/H^+$ antiporter in the renal proximal tubule is evidenced by estimates that this transporter mediates the reabsorption of approximately one third of the filtered load of sodium and the bulk of the secretion of hydrogen ions. This study was designed to investigate the pathway utilized by IGF-I in regulating sodium transport in primary cultured renal proximal tubule cells. Results were as follows : 1. $Na^+$ was observed to accumulate in the primary cells as a function of time. Raising the concentration of extracellular NaCl induced an decrease in $Na^+$ uptake compared with control cells in a dose dependent manner. The rate of $Na^+$ uptake into the primary cells was about two times higher in the absence of NaCl($40.11{\pm}1.76pmole\;Na^+/mg\;protein/min$) than in the presence of 140mM NaCl($17.82{\pm}0.94pmole\;Na^+/mg\;protein/min$) at the 30 minute uptake. 2. $Na^+$ uptake was inhibited by IAA($1{\times}10^{-4}M$) or valinomycin($5{\times}10^{-6}M$) treatment($50.51{\pm}4.04$ and $57.65{\pm}2.27$ of that of control, respectively). $Na^+$ uptake by the primary proximal tubule cells was significantly increased by ouabain($5{\times}10^{-5}M$) treatment($140.23{\pm}3.37%$ of that of control). When actinomycin D($1{\times}10^{-7}M$) or cycloheximide($4{\times}10^{-5}M$) was applied, $Na^+$ uptake was decreased to $90.21{\pm}2.39%$ or $89.64{\pm}3.69%$ of control in IGF-I($1{\times}10^{-5}M$) treated cells, respectively. 3. Extracellular cAMP decreased $Na^+$ uptake in a dose-dependent manner($10^{-8}-10^{-4}M$). IBMX($5{\times}10^{-5}M$) also inhibited $Na^+$ uptake. Treatment of cells with pertussis toxin(50pg/ml) or cholera toxin($1{\mu}g/ml$) inhibited $Na^+$ uptake. Extracellular PMA decreased $Na^+$ uptake in a dose-dependent manner(1-100ng/ml). 100 ng/ml PMA concentration significantly inhibited $Na^+$ uptake in IGF-I treated cells. However, staurosporine($1{\times}10^{-7}M$) had no effect on $Na^+$ uptake. When PMA and staurosporine were added together, the inhibition of $Na^+$ uptake was not observed. In conclusion, sodium uptake in primary cultured rabbit renal proximal tubule cells was dependent on membrane potentials and intracellular energy levels. IGF-I stimulates sodium uptake through mechanisms that involve some degree of de novo protein and/or RNA synthesis, and cAMP and/or PKC pathway mediating the action mechanisms of IGF-I.

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Changes of Soil Chemical Properties and Uptake of Salts by the Plants according to the Application of the Food Waste Compost (음식물찌꺼기 퇴비의 시용에 따른 토양의 화학성 변화 밑 작물체내 염류의 흡수)

  • Lee, Sang-Suk;Chang, Ki-Woon
    • Journal of the Korea Organic Resources Recycling Association
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    • v.6 no.2
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    • pp.59-67
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    • 1998
  • This study was conducted to investigate the changes of soil chemical properties and uptake of salts by the plants(Raphanus and Lactuca) according to the application of the food waste compost(FWC), which was produced on a large scale at a pilot plant. Generally, FWC has a high electrical conductivity(EC) and contains much salts, such as Na, K, Ca, and Mg, and because of these when FWC are applied to soils there is a possibility of salt ac-cumulation in soils and growth inhibition on plants. The FWC were applied at the rates of 0, 20, 40, 80, and 160 Mg/ha in the 1/5,000a pots on the basis of dry weight, and the plants were cultivated for 60 days. And then, changes of pH and EC in soils and uptake of salts by the plants were investigated. The results obtained are summarized as follows; 1. After the cultivation of the Raphanus and Lactuca, pH increased and EC decreased in the soils. 2. Uptake rates of Na and K were slightly increased with the application of the FWC. In uptake rate of Ca, Raphanus and Lactuca was decreased, increased, respectively. In Raphanus, uptake rate of Mg was highest with the application of the FWC at 40 Mg/ha, and in Lactuca, continuously increased with application of the FWC.

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