• Title/Summary/Keyword: $K^+$ efflux

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Effect on the Arginine Transport of Mutant MCAT1, Mouse Cationic Aminoacid Transporter (MCAT1의 돌연변이체가 Arginine 통과 능력에 미치는 영향)

  • Kim, Jung-Woo
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.35-41
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    • 1996
  • To find the substrate interacting site of the MCAT1, charged amino acid residues in the transmembrane domain were changed to opposite charged amino acids and studied the arginine uptake, gp70 binding, efflux and protein expression using the Xenopus oocyte expression method. Among the five mutants of MCAT1, the D403K showed the most interesting characteristics, which had normal gp70 binding but low arginine uptake function, that means the normal expression on the membrane but decreased transport function. All mutants except K211E showed decreased arginine efflux, and kinetic study showed decreased Vmax. Together, Clu(403) residue of MCAT1 may show the possible substrate interacting site in the transmembrane domain of MCAT1.

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A study on the soil $CO_2$ Efflux in Quercus acutissima stand at Mt. Bulam urban nature park (불암산 도시자연공원 상수리나무군락의 토양호흡 특성 연구)

  • Kim, Jeong-Seob;Kong, Seok-Jun;Yang, Keum-Chul
    • Korean Journal of Environment and Ecology
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    • v.28 no.6
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    • pp.762-768
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    • 2014
  • The purpose of this study is to analyze the soil $CO_2$ efflux and micro-climate of a preserved forest area located in a Mt. bulam urban nature park Quercus acutissima stand from June 2013 to May 2014. The research showed that the soil and heterotrophic $CO_2$ efflux were $28.14{\pm}7.99$ to $582.47{\pm}318.51$ and $12.32{\pm}8.04$ to $415.71{\pm}159.92mg\;CO_2{\cdot}m^{-2}{\cdot}h^{-1}$, respectively. In addition the seasonal soil $CO_2$ efflux of summer, autumn, winter, spring were 1169.1, 454.81, 72.08 and $494.23g\;CO_2{\cdot}m^{-2}{\cdot}month^{-1}$, respectively. On the other hand, the seasonal heterotrophic $CO_2$ efflux were 526.20, 340.09, 45.13 and $374.9g\;CO_2{\cdot}m^{-2}{\cdot}month^{-1}$, respectively. Moreover, the annual soil and heterotrophic $CO_2$ efflux was found to be 2190.22 and $1286.33g\;CO_2{\cdot}m^{-2}{\cdot}yr^{-1}$, respectively. The exponential function was also utilized for the regression analysis in order to correlate the environmental factors with the soil and heterotrophic $CO_2$ efflux. It was found out that both air and soil temperatures were positively correlated with the soil and heterotrophic $CO_2$ efflux. However, the amount of solar radiation and soil moisture has showed low correlation for both types of $CO_2$ efflux. Contribution of root $CO_2$ efflux to total soil $CO_2$ efflux in this Quercus acutissima stand was 33.60%.

Effect of sodium on transmembrane calcium movement in the cat ileal longitudinal muscle

  • Rho, Young-Jae;Yun, Il;Kang, Jung-Sook
    • Archives of Pharmacal Research
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    • v.10 no.2
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    • pp.80-87
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    • 1987
  • To get a better insight into the exxistence and the role of a Na-Ca exchange mechanism in smooth muscle, the effect of Na substitution with sucrose on tension development, cellular Ca uptake and $^{45}Ca$ efflux was investigated using isolated cat ileal longitudinal muscle strips. Experimental results were summarized as follows;1) Exposure of the cat ileal longitudinal muscle to Na-free solution induced a contraction, and the magnitude of the contraction increased after incubation of the muscle strips with ouabain ($2{\times10^{-}5}$M) for 1hr. 2) Cellular Ca uptake in Na-free solution increased with an increase in Na content of the Na-loading media, and a linear relationship existed between tissue Na content and cellular Ca uptake for 10 min 3) After tissues were equilibrated in PSS containing $^{45}Ca$ for 2hr, cellular Ca uptake decreased with rising the external Na concentration. 4)Removal of medium Na or inhibition of the Na-K pump decreased the rate of $^{45}Ca$ efflux. These results strongly suggested that Na substitution increases cellular Ca uptake and decreases the rate of $^{45}Ca$ efflux via a Na-Ca exchange mechanism.

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Annual Variation of Soil CO2 Efflux in a Broadleaved Deciduous Forest of the Geumsan (Mt.) Long-Term Ecological Research Site (금산장기생태조사지 낙엽활엽수림내 토양이산화탄소 방출량의 연변동)

  • Kim, Choonsig;Lee, Im Kyun;Lim, Jong Hwan;Park, Byung Bae;Chun, Jung Hwa
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.15 no.3
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    • pp.186-190
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    • 2013
  • Soil respiration in forest ecosystems play an important role in global carbon cycle. This study was carried out to determine the annual variation of soil $CO_2$ efflux for 4 years in a broadleaved deciduous forest of the Geumsan (Mt.) Long-Term Ecological Research (GLTER) site in Southern Korea. The soil $CO_2$ efflux in the GLTER site showed annual variations with the fluctuations of annual mean soil temperature, but not with those of soil water content. The annual mean soil $CO_2$ efflux except for winter season was 0.32 g $CO_2\;m^{-2}h^{-1}$ for 2008, 0.40 g $CO_2\;m^{-2}h^{-1}$ for 2009, 0.41 g $CO_2\;m^{-2}h^{-1}$ for 2007, and 0.54 g $CO_2\;m^{-2}h^{-1}$ for 2010. The lowest soil $CO_2$ effluxin 2008 was associated with the lowest soil temperature ($2.0^{\circ}C$) in comparison with those of other years ($13.0-13.5^{\circ}C$). The exponential relationships between monthly soil $CO_2$ efflux and the corresponding soil temperature at the soil depth of 20 cm were significant ($R^2$ = 0.31-0.75, P < 0.05). The results indicate that the annual variation of soil $CO_2$ efflux was attributed to the variations of soil temperature rather than soil water content in the GLTER site.

Azole Resistance Caused by Increased Drug Efflux in Candida glabrata Isolated from the Urinary Tract of a Dog with Diabetes Mellitus

  • Kim, Minchul;Lee, Hyekyung;Hwang, Sun-Young;Lee, Inhyung;Jung, Won Hee
    • Mycobiology
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    • v.45 no.4
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    • pp.426-429
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    • 2017
  • A yeast-like organism was isolated from a urine sample of a 6-year-old neutered male miniature poodle dog with urinary tract infection, diabetes ketoacidosis, and acute pancreatitis. We identified the yeast-like organism to be Candida glabrata and found that this fungus was highly resistant to azole antifungal drugs. To understand the mechanism of azole resistance in this isolate, the sequences and expression levels of the genes involved in drug resistance were analyzed. The results of our analysis showed that increased drug efflux, mediated by overexpression of ATP transporter genes CDR1 and PDH1, is the main cause of azole resistance of the C. glabrata isolated here.

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.

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.

Distribution of Multidrug Efflux Pump Genes in Enterococci spp. Isolated from Bovine Milk Samples and Their Antibiotic Resistance Patterns (원유 시료에서 분리한 장알균속 세균의 다중약물 유출 펌프(Multidrug Efflux Pump) 유전자의 분포도와 항생제 내성 패턴)

  • Kang, SoWon;Lee, SangJin;Choi, SungSook
    • Korean Journal of Microbiology
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    • v.49 no.2
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    • pp.126-130
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    • 2013
  • The major aim of this study was to investigate the distribution of genes that encode multidrug efflux pumps in Enterococci spp. isolates from bovine milk samples and antibiotic resistance patterns of these strains. Of the 245 isolates, 44.1% showed ampicillin resistance, 79.2% showed erythromycin resistance, 76.3% showed tetracycline resistance and 36.3% showed chloramphenicol resistance. In case of vancomycin and ciprofloxacin, all of the isolates were susceptible to these antibiotics. Of the 245 enterococcal isolates, 82.1% have MFS type eme(A) gene, 72.7% have ABC type efr(A) gene, 77.1% have ABC type efr(B) gene, and 71.8% have ABC type lsa gene. In case of Enterococcus faecalis, the original strain for these genes, 92.5% have eme(A), 87.4% have efr(A), 88.4% have efr(B), and 88.4% have lsa. Interestingly, in case of different species of Enterococci, eme(A) was also detected in four strains of E. faecium, seven strains of E. avium, four strains of E. durans and two strains of E. raffinosus. efr(A) was also detected in two strains of E. faecium and two strains of E. durans and efr(B) was also detected in four strains of E. faecium, five strains of E. avium and four strains of E. durans. This means the possibility of co-transfer of resistance genes between Enterococci species in natural environment. These results are the first report describing the presence of same multidrug efflux pumps in different species of Enterococci in Korea.

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.