• Title/Summary/Keyword: efflux

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Effects of phenylephrine-induced PKC activation on Mg2+ release in guinea pig heart and isolated ventricular myocytes (기니픽 심장과 심근세포에서 Phenylephrine에 의한 PKC 활성화가 Mg2+ 유리에 미치는 영향)

  • Chang, Sung-eun;Kang, Hyung-sub;Kim, Jin-sang
    • Korean Journal of Veterinary Research
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    • v.38 no.1
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    • pp.29-42
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    • 1998
  • $Mg^{2+}$ is one of the most abundant divalent cations in mammalian body(0.2~1.0mM) and the important physiological roles are : first, the cofactor of many enzyme activities, second, the regulator of glycolysis and DNA synthesis, third, the important role of bioenergetics by regulating of phosphorylation, fourth, the influence of cardiac metabolism and function. In this work we have investigated the regulation of the $Mg^{2+}$ induced by ${\alpha}_1-adrenoceptor$ stimulation in perfused guinea pig hearts and isolated myocytes. The $Mg^{2+}$ content of the perfusate or the supernatant was measured by atomic absorbance spectrophotometry. The elimination of $Mg^{2+}$ in the medium increased the force of contraction of right ventricular papillary muscles, and the left ventricular pressure. Phenylephrine also enhanced the force of contraction in the presence of $Mg^{2+}-free$ medium. ${\alpha}_1-Agonists$ such as phenylephrine and methoxamine were found to induce $Mg^{2+}$ efflux in both perfused hearts and myocytes. These effects were blocked by prazosin, an ${\alpha}_1-adrenoceptor$ antagonist. The $Mg^{2+}$ influx could also be induced by phenylephrine and R59022, a diacylglycerol kinase inhibitor. In the presence of protein kinase C(PKC) inhibitors, phenylephrine produced an increase in $Mg^{2+}$ efflux from perfused hearts. Furthermore, $Mg^{2+}$ efflux by phenylephrine was amplified by phorbol 12-myristate 13-acetate(PMA). This enhancement of $Mg^{2+}$ efflux by PMA was blocked by prazosin in perfused hearts. By contrast, the $Mg^{2+}$ influx could be induced by verapamil, nifedipine, ryanodine in perfused hearts, but not in myocytes. $W^7$, a $Ca^{2+}$/calmodulin antagonist, completely blocked the phenylephrine-induced $Mg^{2+}$ efflux in perfused hearts. In conclusion, $Mg^{2+}$ is responsible for the cardiac activity associated with ${\alpha}_1-adrenoceptor$ stimulation. The mobilization of $Mg^{2+}$ is decreased or increased by ${\alpha}_1-adrenoceptor$ stimulation in guinea pig hearts. These responses may be related specifically to the respective pathways of signal transduction. A decrease in $Mg^{2+}$ efflux by ${\alpha}_1-adrenoceptor$ stimulation in hearts can be through PKC dependent and intracellular $Ca^{2+}$ levels.

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Analysis of the Fluoroquinolone Antibiotic Resistance Mechanism of Salmonella enterica Isolates

  • Kim, Soo-Young;Lee, Si-Kyung;Park, Myeong-Soo;Na, Hun-Taek
    • Journal of Microbiology and Biotechnology
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    • v.26 no.9
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    • pp.1605-1612
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    • 2016
  • Quinolone-resistant Salmonella strains were isolated from patient samples, and several quinolone-sensitive strains were used to analyze mutations in the quinolone resistance-determining region (QRDR) of gyrA, gyrB, parC, and parE and to screen for plasmid-mediated quinolone resistance. Among the 21 strains that showed resistance to nalidixic acid and ciprofloxacin (MIC 0.125-2.0 μg/ml), 17 strains had a mutation in QRDR codon 87 of gyrA, and 3 strains had a single mutation (Ser83 → Phe). Another cause of resistance, efflux pump regulation, was studied by examining the expression of acrB, ramA, marA, and soxS. Five strains, including Sal-KH1 and Sal-KH2, showed no increase in relative expression in an analysis using the qRT-PCR method (p < 0.05). In order to determine the genes involved in the resistance, the Sal-9 isolate that showed decreased susceptibility and did not contain a mutation in the gyrA QRDR was used to make the STM (MIC 8 μg/ml) and STH (MIC 16 μg/ml) ciprofloxacin-resistant mutants. The gyrA QRDR Asp87 → Gly mutation was identified in both the STM and STH mutants by mutation analysis. qRT-PCR analysis of the efflux transporter acrB of the AcrAB-TolC efflux system showed increased expression levels in both the STM (1.79-fold) and STH (2.0-fold) mutants. In addition, the expression of the transcriptional regulator marA was increased in both the STM (6.35-fold) and STH (21.73-fold) mutants. Moreover, the expression of soxS was increased in the STM (3.41-fold) and STH (10.05-fold) mutants (p < 0.05). Therefore, these results indicate that AcrAB-TolC efflux pump activity and the target site mutation in gyrA are involved in quinolone resistance.

Effects of Salinity, Temperature and Food Type on the Uptake and Elimination Rates of Cd, Cr, and Zn in the Asiatic Clam Corbicula fluminea

  • Lee, Jung-Suk;Lee, Byeong-Gweon
    • Ocean Science Journal
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    • v.40 no.2
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    • pp.79-89
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    • 2005
  • Laboratory radiotracer experiments were conducted to determine assimilation efficiencies (AE) from ingested algal food and oxic sediment particles, uptake rates from the dissolved phase, and the efflux rates of Cd, Cr and Zn in the Asiatic clam Corbicula fluminea. Among three elements, AE from both algal and sediment food was greatest for Cd, followed by Zn and Cr. The AEs of tested elements from algal food (Phaeodactylum tricornutum) were consistently higher than those from sediments at a given salinity and temperature. The influence of salinity (0, 4 and 8 psu) and temperature (5, 13 and $21^{\circ}C$) on the metal AEs was not evident for most tested elements, except Cd AEs from sediment. The rate constant of metal uptake from the dissolved phase $(k_u)$ was greatest for Cd, followed by Zn and Cr in freshwater media. However, in saline water, the $(k_u)$ of Zn were greater than those of Cd. The influx rate of all tested metals increased with temperature. The efflux rate constant was greatest for Cr $(0.02\;d^{-1})$, followed by Zn $(0.010{\sim}0.017\;d^{-1})$ and $Cd\;(0.006\;d^{-1})$. The efflux rate constant for Zn in clam tissues depurated in 0 psu $(0.017\;d^{-1})$ was faster than that in 8 psu $(0.010\;d^{-1})$. Overall results showed that the variation of salinity and temperature in estuarine systems can considerably influence the metal bioaccumulation potential in the estuarine clam C. fluminea. The relatively high Cd accumulation capacity of C. fluminea characterized by the high AE, high dissolved influx rate and low efflux rate, suggested that this clam species can be used as an efficient biomonitor for the Cd contamination in freshwater and estuarine environments.

Crystal Structure of the Regulatory Domain of MexT, a Transcriptional Activator of the MexEF-OprN Efflux Pump in Pseudomonas aeruginosa

  • Kim, Suhyeon;Kim, Songhee H.;Ahn, Jinsook;Jo, Inseong;Lee, Zee-Won;Choi, Sang Ho;Ha, Nam-Chul
    • Molecules and Cells
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    • v.42 no.12
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    • pp.850-857
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    • 2019
  • The Gram-negative opportunistic pathogen, Pseudomonas aeruginosa, has multiple multidrug efflux pumps. MexT, a LysR-type transcriptional regulator, functions as a transcriptional activator of the MexEF-OprN efflux system. MexT consists of an N-terminal DNA-binding domain and a C-terminal regulatory domain (RD). Little is known regarding MexT ligands and its mechanism of activation. We elucidated the crystal structure of the MexT RD at 2.0 Å resolution. The structure comprised two protomer chains in a dimeric arrangement. MexT possessed an arginine-rich region and a hydrophobic patch lined by a variable loop, both of which are putative ligand-binding sites. The three-dimensional structure of MexT provided clues to the interacting ligand structure. A DNase I footprinting assay of full-length MexT identified two MexT-binding sequence in the mexEF-oprN promoter. Our findings enhance the understanding of the regulation of MexT-dependent activation of efflux pumps.

Analysis of Soil CO2 efflux across three age classes of plantation Pinus koraiensis (임령이 다른 잣나무림에서의 토양 호흡 분석)

  • Nam, Ki-Jung
    • Journal of Wetlands Research
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    • v.20 no.2
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    • pp.116-123
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    • 2018
  • The objective of this study was to examine effects of stand age on soil $CO_2$ efflux in plantation Pinus koraiensis, and to elucidate what extent plant (fine) root and soil microbial biomass contribute to the whole soil $CO_2$ efflux. In three age classes (20-yr-old. 40-yr-old, 70-yr-old) of plantation Pinus koraiensis, in-situ soil respiration, plant fine root biomass and soil microbial biomass were measured from April to November in 2004. Regardless of stand age, soil temperature and soil $CO_2$ efflux increased until July then slowly decreased. Soil respiration was higher in 70-yr-old stand than in 20- and 40-yr stands. Fine root biomass and soil microbial biomass was also higher in 70-yr-old stand. Root exclusion decreased soil respiration in 40-yr stand, but not in 70-yr stand. Soil microbial biomass was higher in 70-yr stand, but there was no monthly variation between July and November. The results suggest that soil respiration may increase as plant stand ages and microbial contribution could play more roles in older stands.

Regulation of Magnesium Release by cAMP during Chemical Hypoxia in the Rat Heart and Isolated Ventricular Myocytes

  • Kim, Jin-Shang;Scarpa, Antonio
    • The Korean Journal of Physiology and Pharmacology
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    • v.3 no.1
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    • pp.59-68
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    • 1999
  • Chemically induced hypoxia has been shown to induce a depletion of ATP. Since intracellular free $Mg^{2+}\;([Mg^{2+}]_i)$ appears to be tightly regulated following cellular energy depletion, we hypothesized that the increase in $[Mg^{2+}]_i$ would result in $Mg^{2+}$ extrusion following hormonal stimulation. To determine the relation between $Mg^{2+}$ efflux and cellular energy state in a hypoxic rat heart and isolated myocytes, $[Mg^{2+}]_i,$ ATP and $Mg^{2+}$ content were measured by using mag-fura-2, luciferin-luciferase and atomic absorbance spectrophotometry. $Mg^{2+}$ effluxes were stimulated by norepinephrine (NE) or cAMP analogues, respectively. $Mg^{2+}$ effluxes induced by NE or cAMP were more stimulated in the presence of metabolic inhibitors (MI). Chemical hypoxia with NaCN (2 mM) caused a rapid decrease of cellular ATP within 1 min. Measurement of $[Mg^{2+}]_i$ confirmed that ATP depletion was accompanied by an increase in $[Mg^{2+}]_i.$ No change in $Mg^{2+}$ efflux was observed when cells were incubated with MI. In the presence of MI, the cAMP-induced $Mg^{2+}$ effluxes were inhibited by quinidine, imipramine, and removal of extracellular $Na^+.$ In addition, after several min of perfusion with $Na^+-free$ buffer, a large increase in $Mg^{2+}$ efflux occurred when $Na^+-free$ buffer was switched to 120 mM $Na^+$ containing buffer. A similar $Mg^{2+}$ efflux was observed in myocytes. These effluxes were inhibited by quinidine and imipramine. These results indicate that the activation of $Mg^{2+}$ effluxes by hormonal stimulation is directly dependent on intracellular $Mg^{2+}$ contents and that these $Mg^{2+}$ effluxes appear to occur through the $Na^+-dependent\;Na^+/Mg^{2+}$ exchange system during chemical hypoxia.

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Valuable Organic Liquid Fertilizer Manufacturing through $TAO^{TM}$ Process for Swine Manure Treatment

  • Lee, Myung-Gyu;Cha, Gi-Cheol
    • Journal of Animal Environmental Science
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    • v.9 no.1
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    • pp.45-56
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    • 2003
  • $TAO^{TM}$ System is an auto-heated thermophilic aerated digestion process using a proprietary microbe called as a Phototropic Bacteria (PTB). High metabolic activity results in heat generation, which enables to produce a pathogen-free and digested liquid fertilizer at short retention times. TAO$^{TM}$ system has been developed to reduce a manure volume and convert into the liquid fertilizer using swine manure since 1992. About 100 units have been installed and operated in Korean swine farms so far. TAO$^{TM}$ system consists of a reactor vessel and ejector-type aeration pumps and foam removers. The swine slurry manure enters into vessel with PTB and is mixed and aerated. The process is operated at detention times from 2 to 4 days and temperature of 55 to $65^{\circ}C$. Foams are occurred and broken down by foam removers to evaporate water contents. Generally, at least 30% of water content is evaporated, 99% of volatile fatty acids caused an odor are removed and pathogen destruction is excellent with fecal coliform, rotavirus and salmonella below detection limits. The effluent from TAO$^{TM}$ system, called as the "TAO EFFLUX", is screened and has superb properties as a fertilizer. Normally N-P-K contents of screened TAO Efflux are 4.7 g/L, 0.375 g/L and 2.8 g/L respectively. The fertilizer effect of TAO EFFLUX compared to chemical fertilizer has been demonstrated and studied with various crops such as rice, potato, cabbage, pumpkin, green pepper, parsley, cucumber and apple. Generally it has better fertilizer effects and excellent soil fertility improvement effects. Moreover, the TAO EFFLUX is concentrated through membrane technology without fouling problems for a cost saving of long distance transportation and a commercialization (crop nutrient commodity) to a gardening market, for example.

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Effect of onion and beet on plasma and liver lipids, platelet aggregation, and erythrocyte Na efflux in simvastatin treated hypercholesterolmic rats

  • Kim, Jung-Lye;Chae, In-Sook;Kang, Young-Hee;Kang, Jung-Sook
    • Nutrition Research and Practice
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    • v.2 no.4
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    • pp.211-217
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    • 2008
  • This study was purposed to investigate the effect of onion or beet on plasma and liver lipids, erythrocyte Na efflux channels and platelet aggregation in simvastatin (SIM) treated hypercholesterolemic rats. Forty Sprague Dawley rats were divided into four groups and fed 0.5% cholesterol based diets containing 2 mg/kg BW simvastatin or simvastatin with 5% onion or beet powder. Plasma total cholesterol was significantly increased in SIM group compared with the control (p<0.01), and the elevated plasma total cholesterol of SIM group was significantly decreased in SIM-onion and SIM-beet groups (p<0.05). HDL-cholesterol in SIM-beet group was significantly increased compared with other groups (p<0.05). Platelet aggregation in both the maximum and initial slope was significantly decreased in SIM group compared with SIM-onion group (p<0.05). Na-K ATPase was significantly decreased in SIM group compared with the control, SIM-onion and SIM-beet groups (p<0.05). Na passive leak was significantly increased in all groups treated with SIM compared with the control (p<0.05). The total Na efflux was decreased in SIM group and increased in SIM-onion group and the difference between these two groups was significant (p<0.05). There was no difference in intracellular Na among groups. In present study, simvastatin, a HMG CoA reductase inhibitor at dose of 2mg/kg BW/day rather increased plasma total cholesterol in rats, inferring that the action mechanism of simvastatin on cholesterol metabolism differ between rat and human. Onion and beet play favorable roles in cardiovascular system by restoring the reduced Na efflux through Na-K ATPase and Na-K cotransport in SIM treated rats.

Effects of Cholesterol Diet and Exercise on Plasma and Liver Lipids, Platelet Aggregation and Erythrocyte Na Efflux in Rats (콜레스테롤 식이와 운동이 흰쥐의 체내 지질수준, 혈소판 응집 및 적혈구막 Na 유출에 미치는 영향)

  • Kang, Min-Sook;Kim, Young-Pyo;Kang, Young-Hee;Kang, Jung-Sook
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.381-390
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    • 2008
  • Present study was conducted to investigate the effects of exercise and cholesterol diet on plasma and liver lipids, platelet aggregation, erythrocyte Na efflux and liver index such as GOT and GPT using Sprague Dawley rats. Forty rats were divided into four groups and fed control or 0.5% cholesterol diet with and without treadmill for six weeks. The final body weight of group fed cholesterol diet with exercise was somewhat decreased compared with group fed cholesterol diet without exercise. L.W/B.W ratio was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise tended to decrease this ratio. Plasma total cholesterol was significantly increased and HDL-cholesterol was decreased in groups fed cholesterol diet (p < 0.01). Plasma triglyceride was significantly decreased in groups fed cholesterol diet compared with groups fed control diet (p < 0.01). Plasma triglyceride of groups with exercise was significantly decreased compared with their non exercising counterparts regardless diet (p < 0.05). Liver total cholesterol and triglyceride was significantly increased in groups fed cholesterol diet (p < 0.01), but exercise did not affect on these levels. Na-K ATPase was somewhat decreased in groups fed cholesterol diet, and exercise tended to recover the reduced Na-K ATPase. Na passive transport was significantly decreased in group fed control diet without exercise and significantly increased in group fed cholesterol diet with exercise, there were significant differences between groups (p < 0.05). There were no differences in total Na efflux and intracellular Na among groups, and total Na efflux was not correlated with intracellular Na. Hematocrit was significantly lower (p < 0.05) in group fed cholesterol diet without exercise compared with other groups. Platelet aggregation in the initial slope and the maximum was increased in groups fed cholesterol diet, but not statistically significant. Exercise especially increased the initial slope of aggregation. Plasma GOT and GPT was significantly increased in groups fed cholesterol diet (p < 0.01), and exercise in group fed cholesterol diet significantly decreased both GOT and GPT compared with the non exercising counterpart (p < 0.01). This study showed that cholesterol diet increases plasma and liver lipids and GOT and GPT, and exercise improves plasma and liver lipid profile and liver index of GOT and GPT preventing fatty liver.

Effects of pH Shock on the Secretion System in Streptomyces coelicolor A3(2)

  • Kim, Yoon-Jung;Song, Jae-Yang;Hong, Soon-Kwang;Smith, Colin P.;Chang, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.658-662
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    • 2008
  • Effects of pH shock on the secretion system of S. coelicolor A3(2) have been investigated at a transcriptional level by using DNA microarrays. Actinorhodin secretion was observed to be highly enhanced when an acidic-pH shock was applied to surface grown cultures of S. coelicolor A3(2). In this culture, a gene of actVA-orf1 encoding a putative efflux pump or transporter protein for actinorhodin was strongly upregulated. A major number of efflux pumps for other metabolites and a major number of secretion proteins for protein secretion were also observed to be upregulated with pH shock. The secretion of actinorhodin was observed to be remarkably enhanced in liquid culture as well.