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#29346565   // To Up

Apigenin induces cell shrinkage in Candida albicans by membrane perturbation.

Apigenin, a natural flavone, has been well characterized for its their anticarcinogenic property; however, its bioactivity against pathogenic fungi has not been investigated in detail. In this study, we examined the antifungal activity and mode of action of apigenin. Apigenin inhibited the growth of fungal pathogens, which induced superficial infection and reduced biofilm mass. Three-dimensional flow cytometric analysis demonstrated that apigenin induced morphological changes, especially cell shrinkage, in Candida albicans. We investigated the cause of cell shrinkage using the cyanine dye 3,3΄-dipropylthiacarbocyanine iodide. Results revealed that apigenin altered the cell membrane potential. Apigenin also induced membrane dysfunction, and increased cell permeability to 1,6-diphenyl-1,3,5-hexatriene and propidium iodide. We observed the influx and efflux of fluorescent molecules of varying molecular weights and radii across large unilamellar vesicles and live cells that had been treated with apigenin. Membrane disruption facilitates the release of small intracellular constituents such as ions and sugars, but not proteins. These findings suggested that apigenin exerted an antifungal activity by inducing membrane disturbances, which led to cell shrinkage and leakage of intracellular components.
Heejeong Lee, Eun-Rhan Woo, Dong Gun Lee

2633 related Products with: Apigenin induces cell shrinkage in Candida albicans by membrane perturbation.

2 Pieces/Box96 samples1 kit10 ug4 Membranes/Box5 mg100ug Lyophilized4 Membranes/Box100 μg1 mL

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#28988287   2017/10/07 To Up

Kinetic Measurements of Di- and Tripeptide and Peptidomimetic Drug Transport in Different Kidney Regions Using the Fluorescent Membrane Potential-Sensitive Dye, DiS-C-(3).

Tri- and dipeptides are transported in the kidney by PEPT1 and PEPT2 isoforms. The aim of this study was to investigate differences in transport kinetics between renal brush border (BBMV) and outer medulla (OMMV) membrane vesicles (where PEPT1 and PEPT2 are sequentially available) for a range of di- and tripeptides and peptidomimetic drugs. This was accomplished through the use of the potential-sensitive fluorescent dye 3,3'-dipropylthiacarbocyanine iodide [DiS-C-(3)]. BBMV and OMMV were prepared from the rat kidney using standard techniques. The presence of PEPT1 in BBMV and PEPT2 in OMMV was confirmed using Western blotting. Fluorescence changes were measured when extravesicular medium at pH 6.6 containing 0-1 mM substrates was added to a cuvette containing vesicles pre-equilibrated at pH 7.4 and 2.71 μM DiS-C-(3). An increase in fluorescence intensity occurred upon substrate addition reflecting the expected positive change in membrane potential difference. Of the range of substrates studied, OMMV manifested the highest affinity to cefadroxil and valacyclovir (K 4.3 ± 1.2 and 11.7 ± 3.2 µM, respectively) compared to other substrates, whilst the BBMV showed a higher affinity to Gly-His (K 15.4 ± 3.1 µM) compared to other substrates. In addition, OMMV showed higher affinity and capacity to Gly-Gln (K 47.1 ± 9.8 µM, 55.5 ± 2.8 ΔF/s/mg protein) than BBMV (K 78.1 ± 13.3 µM and 35.5 ± 1.7 ΔF/s/mg protein, respectively). In conclusion, this study successfully separated the expression of PEPT1 and PEPT2 into different vesicle preparations inferring their activity in different regions of the renal proximal tubule.
Othman A Alghamdi, Nicola King, Graham L Jones, Pierre D J Moens

1300 related Products with: Kinetic Measurements of Di- and Tripeptide and Peptidomimetic Drug Transport in Different Kidney Regions Using the Fluorescent Membrane Potential-Sensitive Dye, DiS-C-(3).

10 mg4 Membranes/Box14 Membranes/Box500 MG10 plates100ul

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#28608233   2017/06/12 To Up

Alleviation by GABA Receptors of Neurotoxicity Mediated by Mitochondrial Permeability Transition Pore in Cultured Murine Cortical Neurons Exposed to N-Methyl-D-aspartate.

Mitochondrial permeability transition pore (PTP) is supposed to at least in part participate in molecular mechanisms underlying the neurotoxicity seen after overactivation of N-methyl-D-aspartate (NMDA) receptor (NMDAR) in neurons. In this study, we have evaluated whether activation of GABA receptor (GABAR), which is linked to membrane G protein-coupled inwardly-rectifying K ion channels (GIRKs), leads to protection of the NMDA-induced neurotoxicity in a manner relevant to mitochondrial membrane depolarization in cultured embryonic mouse cortical neurons. The cationic fluorescent dye 3,3'-dipropylthiacarbocyanine was used for determination of mitochondrial membrane potential. The PTP opener salicylic acid induced a fluorescence increase with a vitality decrease in a manner sensitive to the PTP inhibitor ciclosporin, while ciclosporin alone was effective in significantly preventing both fluorescence increase and viability decrease by NMDA as seen with an NMDAR antagonist. The NMDA-induced fluorescence increase and viability decrease were similarly prevented by pretreatment with the GABAR agonist baclofen, but not by the GABAR agonist muscimol, in a fashion sensitive to a GABAR antagonist. Moreover, the GIRK inhibitor tertiapin canceled the inhibition by baclofen of the NMDA-induced fluorescence increase. These results suggest that GABAR rather than GABAR is protective against the NMDA-induced neurotoxicity mediated by mitochondrial PTP through a mechanism relevant to opening of membrane GIRKs in neurons.
Toshihiko Kinjo, Yoshino Ashida, Hiroshi Higashi, Satoshi Sugimura, Miho Washida, Hiroki Niihara, Kiyokazu Ogita, Yukio Yoneda, Nobuyuki Kuramoto

1391 related Products with: Alleviation by GABA Receptors of Neurotoxicity Mediated by Mitochondrial Permeability Transition Pore in Cultured Murine Cortical Neurons Exposed to N-Methyl-D-aspartate.

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#28007216   2016/09/10 To Up

Antifungal activity of the cationic antimicrobial polymer-polyhexamethylene guanidine hydrochloride and its mode of action.

The antifungal activity of polyhexamethylene guanidine hydrochloride (PHMGH) was studied against various pathogenic fungi. PHMGH had more potent antifungal activity than amphotericin B, which is a commonly used antifungal drug, and also showed no hemolytic and lactate dehydrogenase release activities in the range of 1.25-40.0 μg mL. PHMGH is a cationic polymer containing an amino group and a polymeric guanidine group. Based on its characteristics such as the cationic charge and hydrophobicity, the antifungal mechanism of PHMGH was investigated using Candida albicans, as a model organism. Flow cytometric contour-plot analysis and microscopy showed changes in the size and granularity of the cells after treatment with PHMGH. A membrane study using 1,6-diphenyl-1,3,5-hexatriene labelling indicated a great loss of phospholipid area in the plasma membrane following PHMGH treatment. To investigate the extent of the damage, fluorescein isothiocyanate-labelled dextran leakage from large unilamellar vesicles was observed, indicating that PHMGH acts on the fungal membranes by inducing pore formation, with the majority of pore size being between 2.3 and 3.3 nm. This mechanism was confirmed with ion transition assays using 3,3'-dipropylthiacarbocyanine iodide and an ion-selective electrode meter, which indicated that membrane depolarization involving K leakage was induced. Taken together, these results show that PHMGH exerts its fungicidal effect by forming pores in the cell membrane.
Hyemin Choi, Keuk-Jun Kim, Dong Gun Lee

1045 related Products with: Antifungal activity of the cationic antimicrobial polymer-polyhexamethylene guanidine hydrochloride and its mode of action.

100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized100ug Lyophilized

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#26880798   2016/02/15 To Up

(-)-Nortrachelogenin from Partrinia scabiosaefolia elicits an apoptotic response in Candida albicans.

This study analyzes the antifungal properties of (-)-nortrachelogenin and elucidates its mode of action against pathogenic fungi. We performed susceptibility tests against several pathogenic fungi and verified the absence of hemolysis against human erythrocytes. Its antifungal activity increased reactive oxygen species (ROS) in response to intracellular stress and increased concentrations of both intracellular and extracellular trehalose without causing hemolysis. In addition, a cell wall regeneration study indicated its action on the cytoplasmic membrane. A cell surface study using 3,3(')-dipropylthiacarbocyanine iodide [DiSC3(5)] and 1,6-diphenyl-1,3,5-hexatriene (DPH) demonstrated dissipation of the cytoplasmic membrane at high concentrations. Our study revealed a disturbance in the membrane at higher concentrations and externalization of phosphatidylserine in a dose-dependent manner, affecting other intracellular responses. Furthermore, we investigated the late stage of apoptosis using TUNEL and 4('),6-diamidino-2-phenylindole (DAPI) assays. (-)-Nortrachelogenin-treated cells underwent apoptosis which was triggered by mitochondrial dysfunction via depolarization of the mitochondrial membrane, release of cytochrome c and calcium ion signaling, resulting in the activation of metacaspases. Different concentrations of (-)-nortrachelogenin induced membrane disruption and caspase-dependent apoptosis.
Heejeong Lee, Eun-Rhan Woo, Dong Gun Lee

2121 related Products with: (-)-Nortrachelogenin from Partrinia scabiosaefolia elicits an apoptotic response in Candida albicans.

1 mL100 ug1 mL100ug1 mg100ug 100 UG1 mL100ug Lyophilized100 μg1 Set4 Membranes/Box

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#26514970   2015/10/30 To Up

Effect of extremely low frequency electromagnetic fields on bacterial membrane.

The effect of extremely low frequency electromagnetic fields (ELF-EMF) on bacteria has attracted attention due to its potential for beneficial uses. This research aimed to determine the effect of ELF-EMF on bacterial membrane namely the membrane potential, surface potential, hydrophobicity, respiratory activity and growth.
Sule Oncul, Esra M Cuce, Burak Aksu, Ayse Inhan Garip

1517 related Products with: Effect of extremely low frequency electromagnetic fields on bacterial membrane.

2000 pcs200 ug1 kit 6 ml Ready-to-use 25 ml 4 Membranes/Box2 Membrane supply1 g4 Membranes/Box

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#26420306   2015/09/29 To Up

Antibacterial Mechanism of (-)-Nortrachelogenin in Escherichia coli O157.

(-)-Nortrachelogenin is a lignan belonging to group of polyphenolic compounds. Its biological properties in mammalian cells are well-studied; however, its biological effects in microorganisms remain poorly understood. Its efficacy against pathogenic bacteria, including antibiotic-resistant strains, was investigated and it was found that bacteria are highly susceptible to the antibacterial effects of this compound. To investigate the antibacterial mode of action(s) against Escherichia coli O157, its effect on the penetration of SYTOX green into bacterial cells was assayed. The penetration of SYTOX Green into a bacterial cell is a measure of permeability of the plasma membrane. An increase in fluorescence intensity using bis-(1,3-dibutylbarbituric acid) trimethine oxonol [DiBAC4(3)] and 3,3'-dipropylthiacarbocyanine iodide [DiSC3(5)] was also observed, indicating membrane depolarization. Potassium ion efflux from the cytosol into the extracellular matrix showed that cellular damage due to (-)-nortrachelogenin treatment resulted in the loss of intracellular components. While cells were damaged by (-)-nortrachelogenin, large unilamellar vesicles containing fluorescein isothiocyanate-dextran were perturbed to migrate molecules between 3.3 and 4.8 nm. The release of calcein from giant unilamellar vesicles, occurring as a result of disruption in artificial membranes, was visualized. Taken together, our results indicate that (-)-nortrachelogenin exerts its antibacterial effect by disorganizing and perturbing the cytoplasmic membrane, demonstrating the potential of this compound as a candidate for antibiotic drug development.
Heejeong Lee, Young Rae Ji, Zae Young Ryoo, Myung-Sook Choi, Eun-Rhan Woo, Dong Gun Lee

2195 related Products with: Antibacterial Mechanism of (-)-Nortrachelogenin in Escherichia coli O157.

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#26342880   2015/09/03 To Up

A novel antimicrobial peptide, scolopendin, from Scolopendra subspinipes mutilans and its microbicidal mechanism.

A novel antimicrobial peptide (AMP) was identified from the centipede Scolopendra subspinipes mutilans by RNA sequencing, and the amino acid sequences predicted from the sequenced mRNAs were compared with those of known AMPs. We named this peptide scolopendin, according to its origin, and investigated the molecular mechanisms underlying its antimicrobial activity. Our findings showed that scolopendin had antimicrobial activity against several pathogenic microorganisms, but did not produce hemolysis of human erythrocytes. In addition, disturbances in the cell membrane potential, induction of potassium release from the cytosol, and increased membrane permeability of the microbes Candida albicans and Escherichia coli O157 were detected by the use of 3,3'-dipropylthiacarbocyanine iodide [DiSC3(5)] dye, potassium leakage assay, and propidium iodide influx assay, respectively, following scolopendin treatment. Further evidence to support the membrane-targeted action of scolopendin was obtained using artificial liposomes as models of the cell membrane. Use of calcein and FITC-labeled dextran leakage assays from scolopendin-treated giant unilamellar vesicles and large unilamellar vesicles showed that scolopendin has a pore-forming action on microbial membrane, with an estimated pore radius of 2.3-3.3 nm. In conclusion, scolopendin is a novel and potent AMP with a membrane-targeted mechanism of action.
Wonyoung Lee, Jae-Sam Hwang, Dong Gun Lee

2326 related Products with: A novel antimicrobial peptide, scolopendin, from Scolopendra subspinipes mutilans and its microbicidal mechanism.

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#25863426   2015/04/08 To Up

Single-centred experience with levosimendan in paediatric decompensated dilated cardiomyopathy.

Children with dilated cardiomyopathy in advanced heart failure may spend a long time awaiting heart transplantation. Consequently, mechanical circulatory support is sometimes required as a bridge to transplantation. Levosimendan, a positive inotropic agent, has been reported to be safe and efficient for the treatment of paediatric heart failure.
Pierre-Emmanuel Séguéla, Philippe Mauriat, Jean-Baptiste Mouton, Nadir Tafer, Jana Assy, Géraldine Poncelet, Karine Nubret, Xavier Iriart, Jean-Benoit Thambo

2696 related Products with: Single-centred experience with levosimendan in paediatric decompensated dilated cardiomyopathy.

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#25806026   2015/03/09 To Up

Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence.

Candida albicans is a major cause of opportunistic and life-threatening, systemic fungal infections. Hence new antifungal agents, as well as new methods to treat fungal infections, are still needed. The application of inhibitors of drug-efflux pumps may increase the susceptibility of C. albicans to drugs. We developed a new fluorescence method that allows the in vivo activity evaluation of compounds inhibiting of C. albicans transporters. We show that the potentiometric dye 3,3'-dipropylthiacarbocyanine iodide diS-C3(3) is pumped out by both Cdr1 and Cdr2 transporters. The fluorescence labeling with diS-C3(3) enables a real-time observation of the activity of C. albicans Cdr1 and Cdr2 transporters. We demonstrate that enniatin A and beauvericin show different specificities toward these transporters. Enniatin A inhibits diS-C3(3) efflux by Cdr1 while beauvericin inhibits both Cdr1p and Cdr2p.
Joanna Szczepaniak, Marcin Łukaszewicz, Anna Krasowska

2134 related Products with: Detection of inhibitors of Candida albicans Cdr transporters using a diS-C3(3) fluorescence.

1 mL1 mL1 mL100 ug1 mg1 kit1 kit1 ml10 plates1 kit1 kit

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