• Title/Summary/Keyword: efflux

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Incidence of Erythromycin Resistance Genes, erm(B) and mef(A), in Streptococci Isolated from Dental Plaques of Koreans

  • Kim, Yeon-Hee;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.38 no.2
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    • pp.61-65
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    • 2013
  • Erythromycin is a macrolide antibiotic and inhibits bacterial protein synthesis by stimulating the dissociation of the peptidyl-tRNA molecule from the ribosomes during elongation. The use of macrolides has increased dramatically over the last few years and has led to an increase in bacterial resistance to these antibiotics. Bacterial resistance to erythromycin is generally conferred by the ribosome methylation and/or transport (efflux) protein genes. Among the identified erythromycin-resistant genes, erm(B) (erythromycin methylation) and mef(A) (macrolide efflux) are generally detectable in erythromycin-resistant streptococcal species. The distribution of these genes in oral streptococcal isolates has been reported in studies from other countries but has not been previously examined in a Korean study. We here examined by PCR the presence of erm(B) and mef(A) in oral streptococci isolated from Korean dental plaques. Among the 57 erythromycin-resistant strains tested, 64.9% harbored erm(B) whereas 40.4% were positive for mef(A). Eleven isolates had both the erm(B) and mef(A) genes. Twenty six isolates had only erm(B) and 12 isolates had only mef(A). Eight of the 57 strains examined were negative for both genes.

C-terminal truncated HBx reduces doxorubicin cytotoxicity via ABCB1 upregulation in Huh-7 hepatocellular carcinoma cells

  • Jegal, Myeong-Eun;Jung, Seung-Youn;Han, Yu-Seon;Kim, Yung-Jin
    • BMB Reports
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    • v.52 no.5
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    • pp.330-335
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    • 2019
  • Hepatitis B virus (HBV) encoding the HBV x protein (HBx) is a known causative agent of hepatocellular carcinoma (HCC). Its pathogenic activities in HCC include interference with several signaling pathways associated with cell proliferation and apoptosis. Mutant C-terminal-truncated HBx isoforms are frequently found in human HCC and have been shown to enhance proliferation and invasiveness leading to HCC malignancy. We investigated the molecular mechanism of the reduced doxorubicin cytotoxicity by C-terminal truncated HBx. Cells transfected with C-terminal truncated HBx exhibited reduced cytotoxicity to doxorubicin compared to those transfected with full-length HBx. The doxorubicin resistance of cells expressing C-terminal truncated HBx correlated with upregulation of the ATP binding cassette subfamily B member 1(ABCB1) transporter, resulting in the enhanced efflux of doxorubicin. Inhibiting the activity of ABCB1 and silencing ABCB1 expression by small interfering ribonucleic acid (siRNA) increased the cytotoxicity of doxorubicin. These results indicate that elevated ABCB1 expression induced by C-terminal truncation of HBx was responsible for doxorubicin resistance in HCC. Hence, co-treatment with an ABCB1 inhibitor and an anticancer agent may be effective for the treatment of patients with liver cancer containing the C-terminal truncated HBx.

Effects of Phosphodiesterase 5 Inhibition with Sildenafil on Atrial Contractile and Secretory Function

  • Quan, He Xiu;Kim, Sun-Young;Jin, Xuan-Shun;Park, Jong-Kwan;Kim, Sung-Zoo;Cho, Kyung-Woo
    • The Korean Journal of Physiology and Pharmacology
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    • v.10 no.3
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    • pp.149-154
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    • 2006
  • Selective inhibition of phosphodiesterase (PDE) 5 opened a new therapeutic approach for cardiovascular disorders. Therefore, the effect of PDE5 inhibition on the cardiac function should thoroughly be defined. The purpose of the present study was to define the effects of sildenafil, a selective inhibitor of PDE5, on the atrial cGMP efflux, atrial dynamics, and the release of atrial natriuretic peptide (ANP). By perfusing rabbit left atria to allow atrial pacing, changes in atrial stroke volume and pulse pressure, transmural extracellular fluid translocation, cGMP efflux, and ANP secretion were measured. SIN-I, an NO donor and soluble (s) guanylyl cyclase (GC) activator, and C-type natriuretic peptide (CNP), an activator of particulate (p) GC activator, were used. Sildenafil increased basal levels of cGMP efflux slightly but not significantly. Sildenafil in a therapeutic dose increased atrial dynamics (for atrial stroke volume, $2.84{\pm}1.71%$, n=12, vs $-0.71{\pm}0.86%$, n=21; p<0.05) and decreased ANP release ($-9.02{\pm}3.36%$, n=14, vs $1.35{\pm}3.25%$, n=23; p < 0.05), however, it had no effect on the SIN-1- or CNP-induced increase of cGMP levels. Furthermore, sildenafil in a therapeutic dose accentuated SIN-1-induced, but not CNP-induced, decrease of atrial pulse pressure and ANP release. These data indicate that PDE5 inhibition with sildenafil has a minor effect on cGMP levels, but has a distinct effect on pGC-cGMP- and sGC-cGMP-induced contractile and secretory function.

Hydrogen peroxide inhibits Ca2+ efflux through plasma membrane Ca2+-ATPase in mouse parotid acinar cells

  • Kim, Min Jae;Choi, Kyung Jin;Yoon, Mi Na;Oh, Sang Hwan;Kim, Dong Kwan;Kim, Se Hoon;Park, Hyung Seo
    • The Korean Journal of Physiology and Pharmacology
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    • v.22 no.2
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    • pp.215-223
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    • 2018
  • Intracellular $Ca^{2+}$ mobilization is closely linked with the initiation of salivary secretion in parotid acinar cells. Reactive oxygen species (ROS) are known to be related to a variety of oxidative stress-induced cellular disorders and believed to be involved in salivary impairments. In this study, we investigated the underlying mechanism of hydrogen peroxide ($H_2O_2$) on cytosolic $Ca^{2+}$ accumulation in mouse parotid acinar cells. Intracellular $Ca^{2+}$ levels were slowly elevated when $1mM\;H_2O_2$ was perfused in the presence of normal extracellular $Ca^{2+}$. In a $Ca^{2+}-free$ medium, $1mM\;H_2O_2$ still enhanced the intracellular $Ca^{2+}$ level. $Ca^{2+}$ entry tested using manganese quenching technique was not affected by perfusion of $1mM\;H_2O_2$. On the other hand, $10mM\;H_2O_2$ induced more rapid $Ca^{2+}$ accumulation and facilitated $Ca^{2+}$ entry from extracellular fluid. $Ca^{2+}$ refill into intracellular $Ca^{2+}$ store and inositol 1,4,5-trisphosphate ($1{\mu}M$)-induced $Ca^{2+}$ release from $Ca^{2+}$ store was not affected by $1mM\;H_2O_2$ in permeabilized cells. $Ca^{2+}$ efflux through plasma membrane $Ca^{2+}-ATPase$ (PMCA) was markedly blocked by $1mM\;H_2O_2$ in thapsigargin-treated intact acinar cells. Antioxidants, either catalase or dithiothreitol, completely protected $H_2O_2-induced$ $Ca^{2+}$ accumulation through PMCA inactivation. From the above results, we suggest that excessive production of $H_2O_2$ under pathological conditions may lead to cytosolic $Ca^{2+}$ accumulation and that the primary mechanism of $H_2O_2-induced$ $Ca^{2+}$ accumulation is likely to inhibit $Ca^{2+}$ efflux through PMCA rather than mobilize $Ca^{2+}$ ions from extracellular medium or intracellular stores in mouse parotid acinar cells.

Effect of Gentamicin on Sodium Transport in Human Erythrocytes (Gentamicin이 적혈구막을 통한 $Na^+$ 이동에 미치는 영향)

  • Kim, Kyung-Hyo;Park, Kae-Sook;Kim, Hee-Jin;Shin, Ho-Im;An, Mi-Ra;Kang, Bok-Soon
    • The Korean Journal of Physiology
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    • v.23 no.1
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    • pp.23-34
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    • 1989
  • Gentamicin (GM) is a polybasic, aminoglycoside antibiotic used frequently for the treatment of serious gram-negative infections. The major limiting factors in the clinical use of GM as well as other aminoglycoside antibiotics are their nephrotoxicity and ototoxicity. The primary mechanism of cell injury in aminoglycoside toxicity appears to be the disruption of normal membrane function and the inhibition of $Na^{+}-K^{+}$ ATPase activity. There are both indirect and direct evidences which suggests that the effect of aminoglycoside antibiotics on $Na^{+}-K^{+}$ ATPase may explain, or contribute to, their toxicity. It has been shown that aminoglycoside reduce total ATPase activity (Kaku et al., 1973) and $Na^{+}-K^{+}$ ATPase activity (linuma et al., 1967) in the stria vascularis and spiral ligament of the guinea-pig cochlea. Lipsky and Lietman (1980) reported that aminoglycoside antibitoics inhibited the activity of $Na^{+}-K^{+}$ ATPase in microsomal fractions of the cortex and medulla of the guinea-pig kidney, isolated rat renal tubule and human erythrocyte ghosts. The present invstigation was undertaken to elucidate the mechanism of GM on human erythrocytes by examining its effect on $Na^{+}-K^{+}$ ATPase activity, actives sodium and potassium transport across red blood cell and $^{3}H-ouabain$ binding to red blood cell membranes. The results obtained are summarized as follows: 1) CM inhibited significantly both the activity of total ATPase and $Na^{+}-K^{+}$ ATPase at all concentrations tested. 2) GM inhibited active $^{22}Na$ efflux across red blood cell. When ouabain is present, the rate of $^{22}Na$ efflux was completely inhibited. When both GM and ouabain were added, the inhibitory effect of active $^{22}Na$ efflux was more pronounced. 3) Active $^{86}Rb$ influx was inhibited significantly by GM. In the presence of ouabain, the rate of $^{86}Rb$ influx is markedly inhibited. But $^{86}Rb$ influx is not appreciably altered by the presence of both GM and ouabain. 4) In the presence of GM, $^{3}H-ouabain$ binding to red blood cell membrane increased. From the above results, it may be concluded that the inhibition of active sodium and potassium transport across red blood cell by gentamicin appears to be due to the inhibition of $Na^{+}-K^{+}$ ATPase activity and an increase in ouabain binding to red blood cell membranes.

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Prevalence and Molecular Characterization of Quinolone Antibiotic Resistance in Escherichia coli Isolates from Raw Bulk Milk in Gyeonggi-do (원유시료에서 분리한 대장균의 퀴놀론 항생제 내성 기전)

  • Kang, Sowon;Lee, Sangjin;Choi, Sungsook
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.185-190
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    • 2014
  • The aim of this study was to investigate the prevalence of quinolone resistant E. coli from raw bulk milk and to characterize the resistance determinants. In this study, the gyrA, gyrB, parC, and parE quinolone resistance determining regions (QRDR) were sequenced from quinolone resistant E. coli isolates. Also, the presence of plasmid-mediated quinolone resistance (PMQR) and the expression of efflux pump genes based on quantitative real-time PCR (qRT-PCR) were investigated. Of the 487 coliform bacteria, 9 strains showed nalidixic acid resistance, and 6 of the 9 nalidixic acid resistant isolates were also ciprofloxacin resistant. These 9 strains had a single mutation at codon 83 (S83L) in gyrA, 2 of them had double mutations at codon 83 and 87 (S83L and D87N) in gyrA and 3 of the 9 isolates had single mutations at codon 80 (S80I) in parC. None of the 9 isolates harbored PMQR determinants. Compared with wild-type E. coli ATCC 25922, an over-expression of the acrB gene (2.15-5.74 fold), encoding the pump component of the AcrAB-TolC efflux pump was observed in 4 of 6 ciprofloxacin resistant isolates. This study identified the quinolone resistance mechanism of E. coli isolated from raw milk samples in Gyeonggi-do.

Differential Cytotoxic Effects of Jaspine B in Various Cancer Cells (다양한 암세포주에서 Jaspine B의 함암활성 비교)

  • Lee, Jihoon;Choi, Kwangik;Kwon, Mihwa;Lee, Dongjoo;Choi, Min-Koo;Song, Im-Sook
    • Journal of Life Science
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    • v.26 no.12
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    • pp.1392-1399
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    • 2016
  • Jaspine B is an anhydrophytosphingosine that is isolated from a marine sponge. Because of its structural similarity to sphingosine, it shows anti-cancer effects in human carcinomas. Therefore, this study aims to investigate its anti-proliferative effect on various cancer cells and to correlate its association with the intracellular accumulation of Jaspine B in relevant cancer cells. The anti-proliferative effect of Jaspine B in various cancer cells was determined by a cell viability test, and the intracellular concentration of Jaspine B in relevant cancer cells was determined using mass spectrometry coupled with liquid chromatography. The correlation coefficient and p value between the cytotoxicity and the cell accumulation of Jaspine B were determined using SPSS 16.1. The cytotoxicity of Jaspine B varied depending on the type of cancer cell when compared the $EC_{50}$ values of Jaspine B. Breast and melanoma cancer cells were susceptible to Jaspine B, whereas renal carcinoma cells were resistant. The intracellular concentrations of Jaspine B had a reciprocal correlation with the $EC_{50}$ values in the same cells (r = 0.838). The results suggested that the anti-proliferative effect of Jaspine B was associated with the cellular accumulation of this compound. However, Jaspine B was not a substrate for P-glycoprotein and breast cancer resistance protein, as major efflux pumps caused multidrug resistance. The maintenance of a high intracellular concentration is crucial for the cytotoxic effect of Jaspine B; however, efflux pumps may not be a controlling factor for Jaspine B-related resistance in cancer cells.

The Lipid Efflux Effects of Dichloromethane Extract from Orostachys japonicus in 3T3-L1 Adipocyte Cells (3T3-L1 지방세포에 대한 와송 디클로로메탄 추출물의 지질 대사 개선에 관한 연구)

  • Kim, Soo-Hwan;Lee, Hyeong-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.4
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    • pp.514-520
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    • 2019
  • This study investigated the improved lipid metabolism effect of 3T3-L1 cells induced by adipocytes using the dichloromethane (DCM) fraction in the organic solvent extract of Wassong (Orostachys japonicus). To confirm the cell cytotoxicity, each of 6 fractions of organic solvent extracts (EtOH, Hexane, DCM, EtOAc, BuOH, and H2O) was examined using MTS assay. As a result, it was confirmed that the DCM extract was stable over the whole range of concentrations, and a DCM fraction was used to confirm the improved lipid metabolism effect. Lipid excretion was measured to confirm the change of lipid metabolism. 3T3-L1 cells induced by adipocytes were treated with DCM extract and stained with oil-red O to evaluate lipid accumulation. As a result, it was confirmed that the lipid efflux was significantly improved. In order to confirm the mechanism of lipid efflux, the mRNA expressions of ABCA1 and ABCG1, which are lipid transport proteins, were confirmed by real-time PCR. Therefore, the present study confirmed that the DCM extract from Orostachys japonicus has the effect of improving the lipid metabolism on 3T3-L1 adipocytes. In addition, the results of this study will be used as the basis for the development of functional foods using Orostachys japonicus and also for conducting research on the detailed mechanisms.

Carbon Budget of Pine Forest in Serpentine Area (사문암 지역 소나무림의 탄소수지 연구)

  • Yang, Keum-Chul;Namkung, Hyunmin;Kim, Jeong-Seob;Han, Mi-Kyoung;Shim, Jae-Kuk
    • Korean Journal of Environment and Ecology
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    • v.32 no.6
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    • pp.676-685
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    • 2018
  • This study is to compare carbon budget between serpentine sites and non-serpentine sites dominated by Pinus densiflora forest in the Andong serpentine area where has high values of magnesium and low values of calcium, and are usually deficient in nitrogen and phosphorus, but rich in heavy metals such as nickel, chrome, cobalt, etc. and to measure soil $CO_2$ efflux and environmental factors between January 2017 and December 2017. Soil $CO_2$ efflux was measured with LI-6400 once a month; the soil temperature at 10 cm depth, air temperature, soil moisture contents, and solar radiation were measured in continuum. Soil $CO_2$ efflux in the serpentine area and non-serpentine were $151.71{\pm}75.09g\;CO_2{\cdot}m^{-2}month^{-1}$(42.48 ~ 262.61 g $CO_2{\cdot}m^{-2}month^{-1}$) and $165.09{\pm}118.96g\;CO_2{\cdot}m^{-2}month^{-1}$(20.94 ~ 449.24 g $CO_2{\cdot}m^{-2}month^{-1}$), respectively. Carbon storage in the serpentine area and non-serpentine area were 91.90, $222.85ton{\cdot}ha^{-1}$, respectively. Carbon absorption in the serpentine area and non-serpentine area were 7.99, $17.41ton{\cdot}ha^{-1}yr^{-1}$, respectively. Carbon budget in the serpentine area and non-serpentine area were absorbs 5.3, $14.49ton{\cdot}Cha^{-1}yr^{-1}$, respectively.

Nitrous oxide and carbon dioxide efflux of cropland soil during fallow season (휴경기간 녹비재배 농경지 토양에서 아산화질소 및 이산화탄소 배출특성)

  • Lee, Sun-Il;Kim, Gun-Yeob;Choi, Eun-Jung;Lee, Jong-Sik;Jeong, Hyun-Cheol
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.20 no.4
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    • pp.386-396
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    • 2018
  • Cropland is sources of atmospheric nitrous oxide ($N_2O$) and carbon dioxide ($CO_2$). However, the contribution of the fallow season to emission of these gases has rarely been determined. In this study, a field experiment encompassing three treatments was conducted to determine efflux of $N_2O$ and $CO_2$ in cropland during fallow season. The treatments were hairy vetch (H.V.), rye and control (Con.). The H.V. and rye were sown in middle October and early November, respectively. The soil $N_2O$ efflux among all three treatments in the fallow season (November-April) were $0.014-2.956mg\;N_2O\;m^{-2}{\cdot}d^{-1}$. The cumulative $N_2O$ emissions were $104.4mg\;N_2O\;m^{-2}$ for Con., $85.8mg\;N_2O\;m^{-2}$ for H.V. and $85.0mg\;N_2O\;m^{-2}$ for Rye during the fallow season. The highest $N_2O$ emissions occurred in Con., while H.V. and Rye emissions were similar. Cumulative $CO_2$ emissions were $293.1g\;CO_2\;m^{-2}$ for Con., $242.2g\;CO_2\;m^{-2}$ for H.V., $275.2g\;CO_2\;m^{-2}$ for Rye during fallow season. This study showed that soil $N_2O$ and $CO_2$ average daily emission during fallow season were 28.3% and 27.4%, respectively of the growing season. Our results indicate that $CO_2$ and $N_2O$ emissions from agricultural systems continue throughout the fallow season.