amiloride Millipore Search Results


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  • 95
    Millipore amiloride hydrochloride hydrate
    AP301 activation of ENaC requires pore-forming α - or δ -subunit coexpression. (A) <t>Amiloride</t> (10 µ M)-sensitive current in different subunits and subunit combinations expressed in HEK-293 cells. Cells were patched in the whole-cell
    Amiloride Hydrochloride Hydrate, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 104 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore amiloride
    Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of <t>amiloride</t> (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.
    Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1415 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore powdered amiloride
    Averaged HCO 3 − -independent conductances in the presence and absence of Na + ( A ), DIDS ( B ), tenidap ( C ) and <t>amiloride</t> ( D ) for H 2 O-injected oocytes, oocytes expressing A WT and oocytes expressing A A799V
    Powdered Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Merck KGaA amiloride hydrochloride
    Averaged HCO 3 − -independent conductances in the presence and absence of Na + ( A ), DIDS ( B ), tenidap ( C ) and <t>amiloride</t> ( D ) for H 2 O-injected oocytes, oocytes expressing A WT and oocytes expressing A A799V
    Amiloride Hydrochloride, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 81/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Millipore benzyl amiloride
    Inhibition of Cl − but not Na + transport restores HSV-1 replication in the presence of NaCl: HeLa cells were treated for 24 hours with increasing concentrations of ralfinamide ( a , b ) a voltage gated sodium channel blocker; or benzyl <t>amiloride</t> (c,d) an epithelial sodium channel blocker or ( e,f ) 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) a chloride channel blocker and infected with HSV-1-GFP at a MOI of 0.5. After 1 hour the inoculum was removed and replaced with increasing concentrations of inhibitor and either 0 mM (blue) or 50 mM NaCl (grey). Virus replication was monitored as a function of GFP fluorescence over time and normalised to the no salt control for each channel blocker concentration. Error bars represent the standard error of the mean of multiple replicates. ( b , d , f ) HeLa cells were treated with increasing concentrations of ralfinamide ( b ), benzyl amiloride ( d ) or NPPB ( f ) with 0 mM or 50 mM NaCl. After 24 and 48 hours, cell viability was determined and normalised to untreated cells (0 mM NaCl, 0 mM inhibitor). Error bars represent the standard error of the mean of three experiments carried out in triplicate. **p
    Benzyl Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 84/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore amiloride amd
    Inhibition of Cl − but not Na + transport restores HSV-1 replication in the presence of NaCl: HeLa cells were treated for 24 hours with increasing concentrations of ralfinamide ( a , b ) a voltage gated sodium channel blocker; or benzyl <t>amiloride</t> (c,d) an epithelial sodium channel blocker or ( e,f ) 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) a chloride channel blocker and infected with HSV-1-GFP at a MOI of 0.5. After 1 hour the inoculum was removed and replaced with increasing concentrations of inhibitor and either 0 mM (blue) or 50 mM NaCl (grey). Virus replication was monitored as a function of GFP fluorescence over time and normalised to the no salt control for each channel blocker concentration. Error bars represent the standard error of the mean of multiple replicates. ( b , d , f ) HeLa cells were treated with increasing concentrations of ralfinamide ( b ), benzyl amiloride ( d ) or NPPB ( f ) with 0 mM or 50 mM NaCl. After 24 and 48 hours, cell viability was determined and normalised to untreated cells (0 mM NaCl, 0 mM inhibitor). Error bars represent the standard error of the mean of three experiments carried out in triplicate. **p
    Amiloride Amd, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore dimethyl amiloride dma
    Effects of F 2 on H 2 O 2 -induced and Ang II-induced NHE activity. Notes: ( A ) Intracellular pH (pH i ) was measured with BCECF in rat ventricular myocytes during exposure to NH 4 Cl, followed by removal of external Na + (to induce an acid load) and reintroduction of Na + (Krebs solution). The rapid removal of NH 4 Cl caused an immediate decrease in pH i . Recovery of pH i did not occur until the cells were perfused with Na + -containing Krebs solution. ( B ) Na + -dependent pH i recovery from acid load was completely blocked when 20 μM <t>DMA</t> was present during Na + -free treatment, as well as during recovery in Na + -containing Krebs solution. ( C ) Exposure to 100 μM H 2 O 2 caused an increase in Na + -dependent recovery of pH i from acid load. ( D ) DMA blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( E ) Pretreatment with U0126 inhibited the H 2 O 2 -induced increase in Na + -dependent recovery of pH i . ( F ) Perfusion with 1 μM F 2 completely blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( G ) Perfusion with F 2 completely blocked Na + -dependent pH i recovery in the presence of 1 nM Ang II. Abbreviations: F 2 , N -n-butyl haloperidol iodide; H 2 O 2 , hydrogen peroxide; Ang, angiotensin; NHE, Na + /H + exchanger; BCECF, 2,7-bis(2-carboxyethyl)-5(6)-carboxyfluorescein-acetoxymethyl; U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -aminophenylmercapto)butadiene; DMA, <t>5-(N,N-dimethyl)-amiloride;</t> Na + , sodium ion ; NH 4 Cl, ammonium chloride.
    Dimethyl Amiloride Dma, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore asic blocker amiloride
    The <t>ASIC</t> channel blocker <t>amiloride</t> inhibits extracellular [H + ] evoked currents potentiated by DTT. A. Representative traces. VSMCs pretreated with 1mM DTT for 4 min were exposed to normal external solution ± 100 μM amiloride for 45 sec
    Asic Blocker Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore ethylisopropyl amiloride eipa
    The <t>ASIC</t> channel blocker <t>amiloride</t> inhibits extracellular [H + ] evoked currents potentiated by DTT. A. Representative traces. VSMCs pretreated with 1mM DTT for 4 min were exposed to normal external solution ± 100 μM amiloride for 45 sec
    Ethylisopropyl Amiloride Eipa, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore enac inhibitor amiloride
    UBC response to NaCl is <t>ENaC</t> dependent and not an osmolarity effect. Urethral epithelial cells of ChAT-eGFP reporter mice were isolated and UBC were identified due to eGFP fluorescence. Experiments were performed during continuous superfusion with Tyrode III buffer containing 145 mM Na + . NaCl, mannitol and <t>amiloride</t> were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. In case of NaCl, concentration changes on top of the baseline of concentrations in Tyrode III are indicated. Thus, the total sodium concentration after addition of 1–150 mM ranged from 146 to 295 mM. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence recorded by confocal laser scanning microscopy, correlating to [Ca 2+ ] i . (A) NaCl evokes an increase in [Ca 2+ ] i ; shown are mean and SEM. (B) Representative recording of changes in Calcium Orange® fluorescence in a single cholinergic (eGFP + ) UBC in response to increasing concentrations of NaCl (blue arrowhead) and mannitol (orange arrowhead). (C) Depicted are peak values after stimulation with mannitol (150 mM, N = 22) and NaCl (1 mM, N = 22; 10 mM, N = 22, 50 mM, N = 24, 100 mM, N = 23; 150 mM, N = 21); * P
    Enac Inhibitor Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore channel asics blocker amiloride
    UBC response to NaCl is <t>ENaC</t> dependent and not an osmolarity effect. Urethral epithelial cells of ChAT-eGFP reporter mice were isolated and UBC were identified due to eGFP fluorescence. Experiments were performed during continuous superfusion with Tyrode III buffer containing 145 mM Na + . NaCl, mannitol and <t>amiloride</t> were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. In case of NaCl, concentration changes on top of the baseline of concentrations in Tyrode III are indicated. Thus, the total sodium concentration after addition of 1–150 mM ranged from 146 to 295 mM. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence recorded by confocal laser scanning microscopy, correlating to [Ca 2+ ] i . (A) NaCl evokes an increase in [Ca 2+ ] i ; shown are mean and SEM. (B) Representative recording of changes in Calcium Orange® fluorescence in a single cholinergic (eGFP + ) UBC in response to increasing concentrations of NaCl (blue arrowhead) and mannitol (orange arrowhead). (C) Depicted are peak values after stimulation with mannitol (150 mM, N = 22) and NaCl (1 mM, N = 22; 10 mM, N = 22, 50 mM, N = 24, 100 mM, N = 23; 150 mM, N = 21); * P
    Channel Asics Blocker Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore n n hexamethylene amiloride hma
    The latency of the first bombykal (BAL)-dependent AP was prolonged by all compounds tested. (A,B) The <t>amiloride</t> <t>HMA</t> was significantly more potent than OLC15 and MIA at both Zeitgebertimes. Box plots with whiskers from 5 to 95 percentiles. Different scales were used with interrupted error bars to improve illustration of differences. Significant differences are indicated by asterisks (exact P-values see Table 2 ; *P
    N N Hexamethylene Amiloride Hma, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Millipore epithelial na channel blocker amiloride
    The latency of the first bombykal (BAL)-dependent AP was prolonged by all compounds tested. (A,B) The <t>amiloride</t> <t>HMA</t> was significantly more potent than OLC15 and MIA at both Zeitgebertimes. Box plots with whiskers from 5 to 95 percentiles. Different scales were used with interrupted error bars to improve illustration of differences. Significant differences are indicated by asterisks (exact P-values see Table 2 ; *P
    Epithelial Na Channel Blocker Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 77/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore dimethylamiloride
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
    Dimethylamiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Millipore amide coupling strategy
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
    Amide Coupling Strategy, supplied by Millipore, used in various techniques. Bioz Stars score: 91/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Millipore amide
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
    Amide, supplied by Millipore, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Millipore nhe1 inhibitor 5 n ethyl n isopropyl amiloride
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
    Nhe1 Inhibitor 5 N Ethyl N Isopropyl Amiloride, supplied by Millipore, used in various techniques. Bioz Stars score: 80/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Millipore sodium amide
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
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    92
    Millipore lithium amide
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
    Lithium Amide, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Millipore novapeg rink amide resin
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
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    90
    Millipore sieber amide resins
    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of <t>dimethylamiloride</t> (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response
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    79
    Merck KGaA amiloride hcl
    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    95
    Millipore mitotempo
    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    75
    Millipore tgr amide resin
    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    77
    Millipore novasyntgr amide resin
    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), <t>amiloride</t> ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.
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    Fig. 1. ( A ) Sequence alignments for TM1 and TM2 regions within different members of the ENaC/Degenerin gene family. The alignment was obtained with the GCG Pileup program. Identical and similar residues are printed white on black and white on gray, respectively. Acid-sensing ion channels (ASICs), brain–liver–intestine <t>amiloride-sensitive</t> Na + channel (BLINaC) and epithelial Na + channel (ENaC) correspond to the rat sequences; MEC4 and UNC105 are C.elegans degenerins and dGNaC1 is a Drosophila gonad-specific amiloride-sensitive Na + channel. Note the low conservation between the sequences of TM1 when compared with TM2. ( B ) Current–voltage relationship for wild-type FaNaC expressed in HEK293 cells, in the presence of 30 µM <t>FMRF-amide.</t> Cells were maintained in the whole-cell configuration and voltage steps were applied from –100 to +100 mV in 20 mV increments. (Circles) Symmetrical conditions (140 mM NaCl, 0 mM KCl pH 7.4) between intracellular and extracellular medium. (Squares) Extracellular medium: 95 mM KCl, 35 mM NaCl; intracellular medium: 140 mM NaCl, 0 mM KCl. (Diamonds) Residual currents in the absence of FMRF-amide. Similar residual currents were observed in the presence of 30 µM FMRF-amide and 100 µM amiloride.
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    Fig. 1. ( A ) Sequence alignments for TM1 and TM2 regions within different members of the ENaC/Degenerin gene family. The alignment was obtained with the GCG Pileup program. Identical and similar residues are printed white on black and white on gray, respectively. Acid-sensing ion channels (ASICs), brain–liver–intestine <t>amiloride-sensitive</t> Na + channel (BLINaC) and epithelial Na + channel (ENaC) correspond to the rat sequences; MEC4 and UNC105 are C.elegans degenerins and dGNaC1 is a Drosophila gonad-specific amiloride-sensitive Na + channel. Note the low conservation between the sequences of TM1 when compared with TM2. ( B ) Current–voltage relationship for wild-type FaNaC expressed in HEK293 cells, in the presence of 30 µM <t>FMRF-amide.</t> Cells were maintained in the whole-cell configuration and voltage steps were applied from –100 to +100 mV in 20 mV increments. (Circles) Symmetrical conditions (140 mM NaCl, 0 mM KCl pH 7.4) between intracellular and extracellular medium. (Squares) Extracellular medium: 95 mM KCl, 35 mM NaCl; intracellular medium: 140 mM NaCl, 0 mM KCl. (Diamonds) Residual currents in the absence of FMRF-amide. Similar residual currents were observed in the presence of 30 µM FMRF-amide and 100 µM amiloride.
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    Image Search Results


    AP301 activation of ENaC requires pore-forming α - or δ -subunit coexpression. (A) Amiloride (10 µ M)-sensitive current in different subunits and subunit combinations expressed in HEK-293 cells. Cells were patched in the whole-cell

    Journal: Molecular Pharmacology

    Article Title: Mechanism of Action of Novel Lung Edema Therapeutic AP301 by Activation of the Epithelial Sodium Channel

    doi: 10.1124/mol.113.089409

    Figure Lengend Snippet: AP301 activation of ENaC requires pore-forming α - or δ -subunit coexpression. (A) Amiloride (10 µ M)-sensitive current in different subunits and subunit combinations expressed in HEK-293 cells. Cells were patched in the whole-cell

    Article Snippet: Both amiloride hydrochloride hydrate and TEA were purchased from Sigma-Aldrich GmbH.

    Techniques: Activation Assay

    Amiloride-sensitive AP301-induced current activation in transiently and endogenously expressed ENaC. (A) AP301 activated the Na + current in transiently expressed αβγ -hENaC in HEK-293 and CHO cells with similar efficacy and potency

    Journal: Molecular Pharmacology

    Article Title: Mechanism of Action of Novel Lung Edema Therapeutic AP301 by Activation of the Epithelial Sodium Channel

    doi: 10.1124/mol.113.089409

    Figure Lengend Snippet: Amiloride-sensitive AP301-induced current activation in transiently and endogenously expressed ENaC. (A) AP301 activated the Na + current in transiently expressed αβγ -hENaC in HEK-293 and CHO cells with similar efficacy and potency

    Article Snippet: Both amiloride hydrochloride hydrate and TEA were purchased from Sigma-Aldrich GmbH.

    Techniques: Activation Assay

    AP301 selectively activates αβγ -hENaC expressed in HEK-293 cells. (A) Mean values of inward currents during the control phase, after sequential addition of l - cis (300 µ M), AP301 (120 nM) and final addition of amiloride

    Journal: Molecular Pharmacology

    Article Title: Mechanism of Action of Novel Lung Edema Therapeutic AP301 by Activation of the Epithelial Sodium Channel

    doi: 10.1124/mol.113.089409

    Figure Lengend Snippet: AP301 selectively activates αβγ -hENaC expressed in HEK-293 cells. (A) Mean values of inward currents during the control phase, after sequential addition of l - cis (300 µ M), AP301 (120 nM) and final addition of amiloride

    Article Snippet: Both amiloride hydrochloride hydrate and TEA were purchased from Sigma-Aldrich GmbH.

    Techniques:

    AP301 induced amiloride-sensitive current in proteolytically cleaved hENaC. (A) Cells were patched in the whole-cell mode; inward current was elicited at −100 mV. AP301 (up to 200 nM) did not induce inward sodium current from hENaC endogenously

    Journal: Molecular Pharmacology

    Article Title: Mechanism of Action of Novel Lung Edema Therapeutic AP301 by Activation of the Epithelial Sodium Channel

    doi: 10.1124/mol.113.089409

    Figure Lengend Snippet: AP301 induced amiloride-sensitive current in proteolytically cleaved hENaC. (A) Cells were patched in the whole-cell mode; inward current was elicited at −100 mV. AP301 (up to 200 nM) did not induce inward sodium current from hENaC endogenously

    Article Snippet: Both amiloride hydrochloride hydrate and TEA were purchased from Sigma-Aldrich GmbH.

    Techniques:

    Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of amiloride (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.

    Journal: PLoS ONE

    Article Title: Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice

    doi: 10.1371/journal.pone.0057317

    Figure Lengend Snippet: Representative V TE recordings. The recordings obtained with the final protocol are shown for WT (A) and cftr -/- B6;129 mice (B). Three phases was observed: baseline V TE after Cl - solution perfusion, V TEAmil. after the addition of amiloride (Amil.) and V TELowCl - after the replacement of Cl - solution with a solution of low Cl - concentration (LowCl - ). The inhibitory effect on Cl - secretion of inhibitor-172 and inhibitor-172 plus DIDS was demonstrated in WT mice.

    Article Snippet: The following solutions were perfused through the second lumen at a flow rate of 1.5 ml/h: (1) Cl- solution for basal measurement, (2) 100 µM amiloride in Cl- solution (Sigma-Aldrich, USA), to block ENaC Na+ absorption (3) a low-Cl- solution (140 mM sodium gluconate, 6 mM potassium gluconate, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2 , 6 mM calcium gluconate, pH adjusted to 7.4 with NaOH) containing amiloride (100 µM) to drive Cl- secretion.

    Techniques: Mouse Assay, Concentration Assay

    Cutoff point determination for nasal potential difference parameters distinguishing between FVB WT and F508del-CFTR mice. Receiver operating characteristics (ROC) curves for (A) baseline potential difference (V TE ), (B) amiloride response (ΔV TEAmil. ) and (C) low Cl - solution response (ΔV TElowCl - ), for WT and F508del-CFTR FVB mice are shown. AUC: area under the curve; 95% CI: 95% confidence interval. Cutoff points were determined by the best positive likelihood ratio of sensitivity/(1-specificity).

    Journal: PLoS ONE

    Article Title: Characterization of Nasal Potential Difference in cftr Knockout and F508del-CFTR Mice

    doi: 10.1371/journal.pone.0057317

    Figure Lengend Snippet: Cutoff point determination for nasal potential difference parameters distinguishing between FVB WT and F508del-CFTR mice. Receiver operating characteristics (ROC) curves for (A) baseline potential difference (V TE ), (B) amiloride response (ΔV TEAmil. ) and (C) low Cl - solution response (ΔV TElowCl - ), for WT and F508del-CFTR FVB mice are shown. AUC: area under the curve; 95% CI: 95% confidence interval. Cutoff points were determined by the best positive likelihood ratio of sensitivity/(1-specificity).

    Article Snippet: The following solutions were perfused through the second lumen at a flow rate of 1.5 ml/h: (1) Cl- solution for basal measurement, (2) 100 µM amiloride in Cl- solution (Sigma-Aldrich, USA), to block ENaC Na+ absorption (3) a low-Cl- solution (140 mM sodium gluconate, 6 mM potassium gluconate, 10 mM HEPES, 10 mM glucose, 1 mM MgCl2 , 6 mM calcium gluconate, pH adjusted to 7.4 with NaOH) containing amiloride (100 µM) to drive Cl- secretion.

    Techniques: Mouse Assay

    Effects of cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor-172 and flufenamic acid (FFA) on ATP-induced I sc in mIMCD-K2 cells. A : representative trace of I sc in response to sequential addition of amiloride (Am; apical addition at

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells

    doi: 10.1152/ajprenal.00709.2010

    Figure Lengend Snippet: Effects of cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor-172 and flufenamic acid (FFA) on ATP-induced I sc in mIMCD-K2 cells. A : representative trace of I sc in response to sequential addition of amiloride (Am; apical addition at

    Article Snippet: The apical side of all cell sheets was treated with 10−5 M amiloride (Sigma, St. Louis, MO), an ENaC inhibitor.

    Techniques:

    Effects of the P2Y 1 receptor agonist MRS2365 and the P2Y 2 receptor agonist thio-UTP on I sc in mIMCD-K2 cells. A : representative trace of I sc in response to sequential addition of the following: Am, 10 −5 M amiloride (apical addition); MRS(b) and

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells

    doi: 10.1152/ajprenal.00709.2010

    Figure Lengend Snippet: Effects of the P2Y 1 receptor agonist MRS2365 and the P2Y 2 receptor agonist thio-UTP on I sc in mIMCD-K2 cells. A : representative trace of I sc in response to sequential addition of the following: Am, 10 −5 M amiloride (apical addition); MRS(b) and

    Article Snippet: The apical side of all cell sheets was treated with 10−5 M amiloride (Sigma, St. Louis, MO), an ENaC inhibitor.

    Techniques:

    Sidedness of ATP-induced short-circuit current ( I sc ) in inner medullary collecting duct (mIMCD)-K2 cells. A : representative trace of I sc ; Am indicates 10 −5 M amiloride added to the apical bath; ATP(b) indicates 10 −5 M ATP added to the

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Activation of P2Y1 and P2Y2 receptors induces chloride secretion via calcium-activated chloride channels in kidney inner medullary collecting duct cells

    doi: 10.1152/ajprenal.00709.2010

    Figure Lengend Snippet: Sidedness of ATP-induced short-circuit current ( I sc ) in inner medullary collecting duct (mIMCD)-K2 cells. A : representative trace of I sc ; Am indicates 10 −5 M amiloride added to the apical bath; ATP(b) indicates 10 −5 M ATP added to the

    Article Snippet: The apical side of all cell sheets was treated with 10−5 M amiloride (Sigma, St. Louis, MO), an ENaC inhibitor.

    Techniques:

    NGF enhances macropinocytosis of dextran but not phagocytosis of beads. (a) Phagocytosis of beads : Primary microglia from CX3CR1‐GFP mice were incubated with 6 µm beads and 10 ng/ml IFNγ, 100 ng/ml NGF, 100 nM PMA, and 1 µg/ml Rho/Rac/Cdc42 activator I for 3 hr. (b) Macropinocytosis of dextran : Primary microglia from CX3CR1‐GFP mice were incubated with 2.5 mg/ml Dextran and 10 ng/ml IFNγ, 50 µM Amiloride, 5 µg/ml Cytochalasin‐D, 100 ng/ml NGF, 100 nM PMA, and 1 µg/ml Rho/Rac/Cdc42 activator I for 3 hr. (mean ± SD , * p

    Journal: Glia

    Article Title: NGF steers microglia toward a neuroprotective phenotype, et al. NGF steers microglia toward a neuroprotective phenotype

    doi: 10.1002/glia.23312

    Figure Lengend Snippet: NGF enhances macropinocytosis of dextran but not phagocytosis of beads. (a) Phagocytosis of beads : Primary microglia from CX3CR1‐GFP mice were incubated with 6 µm beads and 10 ng/ml IFNγ, 100 ng/ml NGF, 100 nM PMA, and 1 µg/ml Rho/Rac/Cdc42 activator I for 3 hr. (b) Macropinocytosis of dextran : Primary microglia from CX3CR1‐GFP mice were incubated with 2.5 mg/ml Dextran and 10 ng/ml IFNγ, 50 µM Amiloride, 5 µg/ml Cytochalasin‐D, 100 ng/ml NGF, 100 nM PMA, and 1 µg/ml Rho/Rac/Cdc42 activator I for 3 hr. (mean ± SD , * p

    Article Snippet: Number 485‐MI), Amiloride 50 µM (Sigma‐Aldrich; #A3085), CytochalasinD 10 µg/ml (Sigma‐Aldrich; #C827).

    Techniques: Mouse Assay, Incubation

    H 2 S decreases Na + /K + -ATPase currents of Xenopus lung epithelia. A ) Representative current trace of a control recording. The apical membrane of the lung epithelium was permeabilized with nystatin (100 µM, apical) in the presence of amiloride (10 µM). This resulted in a current increase. When the current was stable, the Na + /K + -ATPase inhibitor ouabain (1 mM) was applied to the basolateral side. The ouabain-sensitive current fraction (ΔI ouab ) represents the activity of the Na + /K + -ATPase. B ) After permeabilisation with nystatin, NaHS (1 mM) was applied to the apical bath. I SC decreased significantly from 20.83±3.99 µA/cm 2 to 16.17±5.23 µA/cm 2 (n = 6, N = 6, p≤0.05). Subsequently, ouabain-sensitive current fractions were determined. C ) Statistical evaluation of experiments as shown in panels A and B. NaHS significantly inhibited Na + /K + -ATPase activity (ΔI ouab ; n = 6, N = 6, p≤0.05). D ) Without permeabilisation with nystatin, ouabain had only a minor effect on transepithelial ion current in the presence of amiloride. Depicted are mean values of non-permeabilized lung epithelia which have been treated with amiloride (10 µM, apical) followed by ouabain (1 mM, basolateral).

    Journal: PLoS ONE

    Article Title: Actions of Hydrogen Sulfide on Sodium Transport Processes across Native Distal Lung Epithelia (Xenopus laevis)

    doi: 10.1371/journal.pone.0100971

    Figure Lengend Snippet: H 2 S decreases Na + /K + -ATPase currents of Xenopus lung epithelia. A ) Representative current trace of a control recording. The apical membrane of the lung epithelium was permeabilized with nystatin (100 µM, apical) in the presence of amiloride (10 µM). This resulted in a current increase. When the current was stable, the Na + /K + -ATPase inhibitor ouabain (1 mM) was applied to the basolateral side. The ouabain-sensitive current fraction (ΔI ouab ) represents the activity of the Na + /K + -ATPase. B ) After permeabilisation with nystatin, NaHS (1 mM) was applied to the apical bath. I SC decreased significantly from 20.83±3.99 µA/cm 2 to 16.17±5.23 µA/cm 2 (n = 6, N = 6, p≤0.05). Subsequently, ouabain-sensitive current fractions were determined. C ) Statistical evaluation of experiments as shown in panels A and B. NaHS significantly inhibited Na + /K + -ATPase activity (ΔI ouab ; n = 6, N = 6, p≤0.05). D ) Without permeabilisation with nystatin, ouabain had only a minor effect on transepithelial ion current in the presence of amiloride. Depicted are mean values of non-permeabilized lung epithelia which have been treated with amiloride (10 µM, apical) followed by ouabain (1 mM, basolateral).

    Article Snippet: Chemicals Amiloride (Sigma) was used as an inhibitor for ENaC and ouabain (Sigma) in order to block the Na+ /K+ -ATPase.

    Techniques: Activity Assay

    Amiloride attenuates the H 2 S induced current decrease. A ) Representative current trace of a control experiment showing the effect of apical treatment with NaHS (1 mM). B ) Lungs were treated with 1 µM amiloride apically and NaHS (1 mM) was subsequently applied for the same duration as the parallel conducted control experiment as depicted in panel A. C ) Statistical evaluation. Amiloride significantly reduced the NaHS-mediated current decrease (ΔI NaHS ; n = 6, N = 6, p≤0.05).

    Journal: PLoS ONE

    Article Title: Actions of Hydrogen Sulfide on Sodium Transport Processes across Native Distal Lung Epithelia (Xenopus laevis)

    doi: 10.1371/journal.pone.0100971

    Figure Lengend Snippet: Amiloride attenuates the H 2 S induced current decrease. A ) Representative current trace of a control experiment showing the effect of apical treatment with NaHS (1 mM). B ) Lungs were treated with 1 µM amiloride apically and NaHS (1 mM) was subsequently applied for the same duration as the parallel conducted control experiment as depicted in panel A. C ) Statistical evaluation. Amiloride significantly reduced the NaHS-mediated current decrease (ΔI NaHS ; n = 6, N = 6, p≤0.05).

    Article Snippet: Chemicals Amiloride (Sigma) was used as an inhibitor for ENaC and ouabain (Sigma) in order to block the Na+ /K+ -ATPase.

    Techniques:

    Interplay of basolateral K + channels and the Na + /K + -ATPase. A ) Representative current trace of a control recording. The apical membrane of the lung epithelium was permeabilized with nystatin (100 µM, apical) in the presence of amiloride (10 µM). This resulted in a current increase. When the current was stable, the Na + /K + -ATPase inhibitor ouabain (1 mM) was applied to the basolateral side. The ouabain-sensitive current fraction (ΔI ouab ) represents the activity of the Na + /K + -ATPase. B ) After permeabilisation with nystatin, lidocaine (1 mM) was applied to the basolateral bath. This resulted in a decrease of the I SC from 17.20±2.58 µA/cm 2 to 12.00±1.67 µA/cm 2 (n = 5, N = 5, p≤0.05). Subsequently, ouabain-sensitive current fractions were determined. C ) Statistical evaluation of experiments as shown in panels A and B. Lidocaine (lido.) significantly inhibited Na + /K + -ATPase activity (ΔI ouab ; n = 5, N = 5, p≤0.05). D ) Without permeabilisation with nystatin, lidocaine had only a minor effect on transepithelial ion current. Depicted are mean values of non-permeabilized lung epithelia which have been treated with amiloride (10 µM, apical) followed by lidocaine (1 mM, basolateral). E ) The application of NaHS (1 mM) to nystatin-permeabilized lungs which have been pre-treated with lidocaine (1 mM) additionally decreased ion current. Note that the subsequent application of ouabain was without any further effect. F ) NaHS significantly decreased I SC under conditions as shown in panel E (n = 6, N = 6, p≤0.05).

    Journal: PLoS ONE

    Article Title: Actions of Hydrogen Sulfide on Sodium Transport Processes across Native Distal Lung Epithelia (Xenopus laevis)

    doi: 10.1371/journal.pone.0100971

    Figure Lengend Snippet: Interplay of basolateral K + channels and the Na + /K + -ATPase. A ) Representative current trace of a control recording. The apical membrane of the lung epithelium was permeabilized with nystatin (100 µM, apical) in the presence of amiloride (10 µM). This resulted in a current increase. When the current was stable, the Na + /K + -ATPase inhibitor ouabain (1 mM) was applied to the basolateral side. The ouabain-sensitive current fraction (ΔI ouab ) represents the activity of the Na + /K + -ATPase. B ) After permeabilisation with nystatin, lidocaine (1 mM) was applied to the basolateral bath. This resulted in a decrease of the I SC from 17.20±2.58 µA/cm 2 to 12.00±1.67 µA/cm 2 (n = 5, N = 5, p≤0.05). Subsequently, ouabain-sensitive current fractions were determined. C ) Statistical evaluation of experiments as shown in panels A and B. Lidocaine (lido.) significantly inhibited Na + /K + -ATPase activity (ΔI ouab ; n = 5, N = 5, p≤0.05). D ) Without permeabilisation with nystatin, lidocaine had only a minor effect on transepithelial ion current. Depicted are mean values of non-permeabilized lung epithelia which have been treated with amiloride (10 µM, apical) followed by lidocaine (1 mM, basolateral). E ) The application of NaHS (1 mM) to nystatin-permeabilized lungs which have been pre-treated with lidocaine (1 mM) additionally decreased ion current. Note that the subsequent application of ouabain was without any further effect. F ) NaHS significantly decreased I SC under conditions as shown in panel E (n = 6, N = 6, p≤0.05).

    Article Snippet: Chemicals Amiloride (Sigma) was used as an inhibitor for ENaC and ouabain (Sigma) in order to block the Na+ /K+ -ATPase.

    Techniques: Activity Assay

    H 2 S inhibits amiloride-sensitive Na + absorption. A ) Representative current trace of a control experiment. In order to estimate the amount of Na + absorption, the ENaC inhibitor amiloride (10 µM) was applied apically. B ) Similar experiment showing the effects of amiloride after apical pre-treatment of the lung epithelium with apical NaHS (1 mM). C ) Statistical evaluation of experiments as shown in panels A and B. Depicted are amiloride-sensitive (ΔI ami ) current fractions. NaHS significantly reduced ΔI ami from 45.04±5.55 µA/cm 2 to 14.84±1.77 µA/cm 2 (n = 5; N = 3; p≤0.001).

    Journal: PLoS ONE

    Article Title: Actions of Hydrogen Sulfide on Sodium Transport Processes across Native Distal Lung Epithelia (Xenopus laevis)

    doi: 10.1371/journal.pone.0100971

    Figure Lengend Snippet: H 2 S inhibits amiloride-sensitive Na + absorption. A ) Representative current trace of a control experiment. In order to estimate the amount of Na + absorption, the ENaC inhibitor amiloride (10 µM) was applied apically. B ) Similar experiment showing the effects of amiloride after apical pre-treatment of the lung epithelium with apical NaHS (1 mM). C ) Statistical evaluation of experiments as shown in panels A and B. Depicted are amiloride-sensitive (ΔI ami ) current fractions. NaHS significantly reduced ΔI ami from 45.04±5.55 µA/cm 2 to 14.84±1.77 µA/cm 2 (n = 5; N = 3; p≤0.001).

    Article Snippet: Chemicals Amiloride (Sigma) was used as an inhibitor for ENaC and ouabain (Sigma) in order to block the Na+ /K+ -ATPase.

    Techniques:

    Proposed effects of amiloride derivation on the hGABA-(A)ρ1 receptor. When the guanidine group is accessible and the pyrazine ring remains unchanged, as in the case of amiloride, there is positive allosteric modulatory activity and weak direct activity on the hGABA-A ρ1 receptor. The HMA structure is produced from derivation of the pyrazine ring along with accessibility of the guanidine group. If there is a substitution on the guanidine group, however, the resulting derivative fails to elicit positive allosteric modulatory activity from the receptor, as in the case of benzamil and phenamil.

    Journal: Channels

    Article Title: 5-(N, N-Hexamethylene) amiloride is a GABA-A ρ1 receptor positive allosteric modulator

    doi: 10.1080/19336950.2016.1207021

    Figure Lengend Snippet: Proposed effects of amiloride derivation on the hGABA-(A)ρ1 receptor. When the guanidine group is accessible and the pyrazine ring remains unchanged, as in the case of amiloride, there is positive allosteric modulatory activity and weak direct activity on the hGABA-A ρ1 receptor. The HMA structure is produced from derivation of the pyrazine ring along with accessibility of the guanidine group. If there is a substitution on the guanidine group, however, the resulting derivative fails to elicit positive allosteric modulatory activity from the receptor, as in the case of benzamil and phenamil.

    Article Snippet: Amiloride-derivatives were purchased from Sigma-Aldrich.

    Techniques: Activity Assay, Produced

    5-(N,N-Hexamethylene) amiloride (HMA) potentiates the hGABA-(A)ρ1 receptor. (A) Resulting representative wild type hGABA-A ρ1 receptor current generated by the co-application of GABA (10 µM) with increasing concentrations of HMA are shown. (B) Summary of potentiation of the hGABA-A ρ1 receptor in the presence of increasing HMA. Data are presented as the mean ± SEM of n ≥ 4 cells. Significance is indicated by: *, p ≤ 0.05 using a One-way ANOVA comparison with Bonferroni post-hoc test compared to the control. Dashed square highlights the guanidine group.

    Journal: Channels

    Article Title: 5-(N, N-Hexamethylene) amiloride is a GABA-A ρ1 receptor positive allosteric modulator

    doi: 10.1080/19336950.2016.1207021

    Figure Lengend Snippet: 5-(N,N-Hexamethylene) amiloride (HMA) potentiates the hGABA-(A)ρ1 receptor. (A) Resulting representative wild type hGABA-A ρ1 receptor current generated by the co-application of GABA (10 µM) with increasing concentrations of HMA are shown. (B) Summary of potentiation of the hGABA-A ρ1 receptor in the presence of increasing HMA. Data are presented as the mean ± SEM of n ≥ 4 cells. Significance is indicated by: *, p ≤ 0.05 using a One-way ANOVA comparison with Bonferroni post-hoc test compared to the control. Dashed square highlights the guanidine group.

    Article Snippet: Amiloride-derivatives were purchased from Sigma-Aldrich.

    Techniques: Generated

    Selected Ala mutants alter the activity of the human αT663 variant. A : αT663βγ (wild-type), αA664βγ, and αA667βγ channels were expressed in oocytes. Amiloride-sensitive whole

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Multiple residues in the distal C terminus of the ?-subunit have roles in modulating human epithelial sodium channel activity

    doi: 10.1152/ajprenal.00493.2011

    Figure Lengend Snippet: Selected Ala mutants alter the activity of the human αT663 variant. A : αT663βγ (wild-type), αA664βγ, and αA667βγ channels were expressed in oocytes. Amiloride-sensitive whole

    Article Snippet: ENaC-mediated current was defined as the difference in whole oocyte current at either −100- or −60-mV holding potential (adjusted for resting membrane potential) before and after addition of 10 μM amiloride-HCl (Sigma) to the bath solution.

    Techniques: Activity Assay, Variant Assay

    αC664M mutation selectively reduces functional expression of the αT663 variant. αT663 and αA663 variants were expressed in oocytes with or without the αC664M mutation. Amiloride-sensitive whole cell currents were

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Multiple residues in the distal C terminus of the ?-subunit have roles in modulating human epithelial sodium channel activity

    doi: 10.1152/ajprenal.00493.2011

    Figure Lengend Snippet: αC664M mutation selectively reduces functional expression of the αT663 variant. αT663 and αA663 variants were expressed in oocytes with or without the αC664M mutation. Amiloride-sensitive whole cell currents were

    Article Snippet: ENaC-mediated current was defined as the difference in whole oocyte current at either −100- or −60-mV holding potential (adjusted for resting membrane potential) before and after addition of 10 μM amiloride-HCl (Sigma) to the bath solution.

    Techniques: Mutagenesis, Functional Assay, Expressing, Variant Assay

    SP-Adv cellular uptake is independent of CAR-regulated endocytosis. Notes: MSCs (2×10 4 cells/well) were pretreated with or without amiloride, MBCD, or CPA for 1 hour. Cells were then transduced with Advs or SP-Advs encoding luciferase. After culturing for 24 hours, luciferase expression was measured (n=6). All data are represented as mean ± sd (** P

    Journal: International Journal of Nanomedicine

    Article Title: Coating with spermine-pullulan polymer enhances adenoviral transduction of mesenchymal stem cells

    doi: 10.2147/IJN.S109897

    Figure Lengend Snippet: SP-Adv cellular uptake is independent of CAR-regulated endocytosis. Notes: MSCs (2×10 4 cells/well) were pretreated with or without amiloride, MBCD, or CPA for 1 hour. Cells were then transduced with Advs or SP-Advs encoding luciferase. After culturing for 24 hours, luciferase expression was measured (n=6). All data are represented as mean ± sd (** P

    Article Snippet: The following day, the control group was pretreated with PBS, others were pretreated with one of the following endocytosis inhibitors for 1 hour: 6.65 mg/mL methyl-β-cyclodextrin (MBCD) (Kaiyang Bio Co, Shanghai, People’s Republic of China), 10 μg/mL chlorpromazine (CPA) (Kaiyang Bio Co) or 2.5 mM amiloride HCl hydrate (Sigma-Aldrich Co.).

    Techniques: Transduction, Luciferase, Expressing

    Averaged HCO 3 − -independent conductances in the presence and absence of Na + ( A ), DIDS ( B ), tenidap ( C ) and amiloride ( D ) for H 2 O-injected oocytes, oocytes expressing A WT and oocytes expressing A A799V

    Journal: The Journal of Physiology

    Article Title: HCO3\u2212-independent conductance with a mutant Na+/HCO3\u2212 cotransporter (SLC4A4) in a case of proximal renal tubular acidosis with hypokalaemic paralysis

    doi: 10.1113/jphysiol.2011.224733

    Figure Lengend Snippet: Averaged HCO 3 − -independent conductances in the presence and absence of Na + ( A ), DIDS ( B ), tenidap ( C ) and amiloride ( D ) for H 2 O-injected oocytes, oocytes expressing A WT and oocytes expressing A A799V

    Article Snippet: Amiloride-containing solutions were prepared at working concentration by the addition of powdered amiloride (Sigma-Aldrich) to the solution.

    Techniques: Injection, Expressing

    Inhibition of Cl − but not Na + transport restores HSV-1 replication in the presence of NaCl: HeLa cells were treated for 24 hours with increasing concentrations of ralfinamide ( a , b ) a voltage gated sodium channel blocker; or benzyl amiloride (c,d) an epithelial sodium channel blocker or ( e,f ) 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) a chloride channel blocker and infected with HSV-1-GFP at a MOI of 0.5. After 1 hour the inoculum was removed and replaced with increasing concentrations of inhibitor and either 0 mM (blue) or 50 mM NaCl (grey). Virus replication was monitored as a function of GFP fluorescence over time and normalised to the no salt control for each channel blocker concentration. Error bars represent the standard error of the mean of multiple replicates. ( b , d , f ) HeLa cells were treated with increasing concentrations of ralfinamide ( b ), benzyl amiloride ( d ) or NPPB ( f ) with 0 mM or 50 mM NaCl. After 24 and 48 hours, cell viability was determined and normalised to untreated cells (0 mM NaCl, 0 mM inhibitor). Error bars represent the standard error of the mean of three experiments carried out in triplicate. **p

    Journal: Scientific Reports

    Article Title: Antiviral innate immune response in non-myeloid cells is augmented by chloride ions via an increase in intracellular hypochlorous acid levels

    doi: 10.1038/s41598-018-31936-y

    Figure Lengend Snippet: Inhibition of Cl − but not Na + transport restores HSV-1 replication in the presence of NaCl: HeLa cells were treated for 24 hours with increasing concentrations of ralfinamide ( a , b ) a voltage gated sodium channel blocker; or benzyl amiloride (c,d) an epithelial sodium channel blocker or ( e,f ) 5-nitro-2-(3-phenylpropyl-amino) benzoic acid (NPPB) a chloride channel blocker and infected with HSV-1-GFP at a MOI of 0.5. After 1 hour the inoculum was removed and replaced with increasing concentrations of inhibitor and either 0 mM (blue) or 50 mM NaCl (grey). Virus replication was monitored as a function of GFP fluorescence over time and normalised to the no salt control for each channel blocker concentration. Error bars represent the standard error of the mean of multiple replicates. ( b , d , f ) HeLa cells were treated with increasing concentrations of ralfinamide ( b ), benzyl amiloride ( d ) or NPPB ( f ) with 0 mM or 50 mM NaCl. After 24 and 48 hours, cell viability was determined and normalised to untreated cells (0 mM NaCl, 0 mM inhibitor). Error bars represent the standard error of the mean of three experiments carried out in triplicate. **p

    Article Snippet: The following day the medium was removed, and cells were pre-treated with increasing concentrations of sodium/chloride channel inhibitor (ralfinamide (Tocris) − voltage gated sodium channels; benzyl amiloride (Sigma) − epithelial sodium channels; NPPB (Sigma) − chloride channels) or the myeloperoxidase inhibitor 4-Aminobenzoic hydrazide (0, 1, 2.5, 5, 10, 20 or 40 μM) (Abcam).

    Techniques: Inhibition, Infection, Fluorescence, Concentration Assay

    Effects of F 2 on H 2 O 2 -induced and Ang II-induced NHE activity. Notes: ( A ) Intracellular pH (pH i ) was measured with BCECF in rat ventricular myocytes during exposure to NH 4 Cl, followed by removal of external Na + (to induce an acid load) and reintroduction of Na + (Krebs solution). The rapid removal of NH 4 Cl caused an immediate decrease in pH i . Recovery of pH i did not occur until the cells were perfused with Na + -containing Krebs solution. ( B ) Na + -dependent pH i recovery from acid load was completely blocked when 20 μM DMA was present during Na + -free treatment, as well as during recovery in Na + -containing Krebs solution. ( C ) Exposure to 100 μM H 2 O 2 caused an increase in Na + -dependent recovery of pH i from acid load. ( D ) DMA blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( E ) Pretreatment with U0126 inhibited the H 2 O 2 -induced increase in Na + -dependent recovery of pH i . ( F ) Perfusion with 1 μM F 2 completely blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( G ) Perfusion with F 2 completely blocked Na + -dependent pH i recovery in the presence of 1 nM Ang II. Abbreviations: F 2 , N -n-butyl haloperidol iodide; H 2 O 2 , hydrogen peroxide; Ang, angiotensin; NHE, Na + /H + exchanger; BCECF, 2,7-bis(2-carboxyethyl)-5(6)-carboxyfluorescein-acetoxymethyl; U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -aminophenylmercapto)butadiene; DMA, 5-(N,N-dimethyl)-amiloride; Na + , sodium ion ; NH 4 Cl, ammonium chloride.

    Journal: Drug Design, Development and Therapy

    Article Title: N-n-butyl haloperidol iodide inhibits H2O2-induced Na+/Ca2+-exchanger activation via the Na+/H+ exchanger in rat ventricular myocytes

    doi: 10.2147/DDDT.S63163

    Figure Lengend Snippet: Effects of F 2 on H 2 O 2 -induced and Ang II-induced NHE activity. Notes: ( A ) Intracellular pH (pH i ) was measured with BCECF in rat ventricular myocytes during exposure to NH 4 Cl, followed by removal of external Na + (to induce an acid load) and reintroduction of Na + (Krebs solution). The rapid removal of NH 4 Cl caused an immediate decrease in pH i . Recovery of pH i did not occur until the cells were perfused with Na + -containing Krebs solution. ( B ) Na + -dependent pH i recovery from acid load was completely blocked when 20 μM DMA was present during Na + -free treatment, as well as during recovery in Na + -containing Krebs solution. ( C ) Exposure to 100 μM H 2 O 2 caused an increase in Na + -dependent recovery of pH i from acid load. ( D ) DMA blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( E ) Pretreatment with U0126 inhibited the H 2 O 2 -induced increase in Na + -dependent recovery of pH i . ( F ) Perfusion with 1 μM F 2 completely blocked Na + -dependent pH i recovery in the presence of H 2 O 2 . ( G ) Perfusion with F 2 completely blocked Na + -dependent pH i recovery in the presence of 1 nM Ang II. Abbreviations: F 2 , N -n-butyl haloperidol iodide; H 2 O 2 , hydrogen peroxide; Ang, angiotensin; NHE, Na + /H + exchanger; BCECF, 2,7-bis(2-carboxyethyl)-5(6)-carboxyfluorescein-acetoxymethyl; U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -aminophenylmercapto)butadiene; DMA, 5-(N,N-dimethyl)-amiloride; Na + , sodium ion ; NH 4 Cl, ammonium chloride.

    Article Snippet: HEPES (4,[2-hydroxyethyl]-1-piperazine-ethanesulphonic acid]), CsCl, 1,2-bis(2-aminophenoxy)-ethane-N ,N ,N ′,N ′-tetraacetic acid (BAPTA), ouabain, nifedipine, ryanodine, epidermal growth factor (EGF), angiotensin (Ang) II and 5-(N ,N -dimethyl)-amiloride (DMA) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA).

    Techniques: Activity Assay

    Effects of U0126 and DMA on the H 2 O 2 -induced I NCX increase. Notes: ( A ) I – V curves of control (a), or in the presence of 100 μM H 2 O 2 (b), H 2 O 2 + U0126 (5 μM) (c), and 5 mM NiCl 2 (d). ( B ) I – V curves of net Ni 2+ -sensitive currents, obtained by subtracting the corresponding I – V curves in ( A ). ( C ) I – V curves of control (a), or in the presence of 100 μM H 2 O 2 (b), H 2 O 2 + DMA (20 μM) (c), and 5 mM NiCl 2 (d). ( D ) I – V curves of net Ni 2+ -sensitive currents, obtained by subtracting the corresponding I – V curves in ( C ). Abbreviations: U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -aminophenylmercapto)butadiene; DMA, 5-( N , N -dimethyl)-amiloride; H 2 O 2 , hydrogen peroxide; NiCl 2 , nickel chloride; I NCX , current of Na + /Ca 2+ exchanger; NiCl 2 , nickel chloride; I – V , current–voltage; Ni 2+ , nickel ion.

    Journal: Drug Design, Development and Therapy

    Article Title: N-n-butyl haloperidol iodide inhibits H2O2-induced Na+/Ca2+-exchanger activation via the Na+/H+ exchanger in rat ventricular myocytes

    doi: 10.2147/DDDT.S63163

    Figure Lengend Snippet: Effects of U0126 and DMA on the H 2 O 2 -induced I NCX increase. Notes: ( A ) I – V curves of control (a), or in the presence of 100 μM H 2 O 2 (b), H 2 O 2 + U0126 (5 μM) (c), and 5 mM NiCl 2 (d). ( B ) I – V curves of net Ni 2+ -sensitive currents, obtained by subtracting the corresponding I – V curves in ( A ). ( C ) I – V curves of control (a), or in the presence of 100 μM H 2 O 2 (b), H 2 O 2 + DMA (20 μM) (c), and 5 mM NiCl 2 (d). ( D ) I – V curves of net Ni 2+ -sensitive currents, obtained by subtracting the corresponding I – V curves in ( C ). Abbreviations: U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -aminophenylmercapto)butadiene; DMA, 5-( N , N -dimethyl)-amiloride; H 2 O 2 , hydrogen peroxide; NiCl 2 , nickel chloride; I NCX , current of Na + /Ca 2+ exchanger; NiCl 2 , nickel chloride; I – V , current–voltage; Ni 2+ , nickel ion.

    Article Snippet: HEPES (4,[2-hydroxyethyl]-1-piperazine-ethanesulphonic acid]), CsCl, 1,2-bis(2-aminophenoxy)-ethane-N ,N ,N ′,N ′-tetraacetic acid (BAPTA), ouabain, nifedipine, ryanodine, epidermal growth factor (EGF), angiotensin (Ang) II and 5-(N ,N -dimethyl)-amiloride (DMA) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA).

    Techniques:

    Effect of F 2 on NHE and NCX protein expression. Notes: ( A and C ) Western blot analysis of total NHE protein. ( B and D ) Western blot analysis of total NCX protein. Upper, representative blot of three independent experiments; lower, quantitative densitometric data were normalized as a percentage of those of the control group, which was plotted as 100%. Abbreviations: F 2 , N -n-butyl haloperidol iodide; NHE, Na + /H + exchanger; NCX, Na + /Ca 2+ exchanger; Ang, angiotensin; U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -amino-phenyl mercapto)butadiene; DMA, 5-( N,N -dimethyl)-amiloride; H 2 O 2 , hydrogen peroxide; EGF, epidermal growth factor.

    Journal: Drug Design, Development and Therapy

    Article Title: N-n-butyl haloperidol iodide inhibits H2O2-induced Na+/Ca2+-exchanger activation via the Na+/H+ exchanger in rat ventricular myocytes

    doi: 10.2147/DDDT.S63163

    Figure Lengend Snippet: Effect of F 2 on NHE and NCX protein expression. Notes: ( A and C ) Western blot analysis of total NHE protein. ( B and D ) Western blot analysis of total NCX protein. Upper, representative blot of three independent experiments; lower, quantitative densitometric data were normalized as a percentage of those of the control group, which was plotted as 100%. Abbreviations: F 2 , N -n-butyl haloperidol iodide; NHE, Na + /H + exchanger; NCX, Na + /Ca 2+ exchanger; Ang, angiotensin; U0126, 1,4-diamino-2,3-dicyano-1,4-bis( o -amino-phenyl mercapto)butadiene; DMA, 5-( N,N -dimethyl)-amiloride; H 2 O 2 , hydrogen peroxide; EGF, epidermal growth factor.

    Article Snippet: HEPES (4,[2-hydroxyethyl]-1-piperazine-ethanesulphonic acid]), CsCl, 1,2-bis(2-aminophenoxy)-ethane-N ,N ,N ′,N ′-tetraacetic acid (BAPTA), ouabain, nifedipine, ryanodine, epidermal growth factor (EGF), angiotensin (Ang) II and 5-(N ,N -dimethyl)-amiloride (DMA) were purchased from Sigma-Aldrich Co. (St Louis, MO, USA).

    Techniques: Expressing, Western Blot

    The ASIC channel blocker amiloride inhibits extracellular [H + ] evoked currents potentiated by DTT. A. Representative traces. VSMCs pretreated with 1mM DTT for 4 min were exposed to normal external solution ± 100 μM amiloride for 45 sec

    Journal: Cellular Physiology and Biochemistry

    Article Title: ASIC-like Currents in Freshly Isolated Cerebral Artery Smooth Muscle Cells are Inhibited by Endogenous Oxidase Activity

    doi: 10.1159/000325215

    Figure Lengend Snippet: The ASIC channel blocker amiloride inhibits extracellular [H + ] evoked currents potentiated by DTT. A. Representative traces. VSMCs pretreated with 1mM DTT for 4 min were exposed to normal external solution ± 100 μM amiloride for 45 sec

    Article Snippet: Cells were preincubated with the ASIC blocker amiloride (100 μM, Sigma Chemicals) for 45s to confirm DTT potentiated currents were generated by ASIC-like channels.

    Techniques: Size-exclusion Chromatography

    UBC response to NaCl is ENaC dependent and not an osmolarity effect. Urethral epithelial cells of ChAT-eGFP reporter mice were isolated and UBC were identified due to eGFP fluorescence. Experiments were performed during continuous superfusion with Tyrode III buffer containing 145 mM Na + . NaCl, mannitol and amiloride were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. In case of NaCl, concentration changes on top of the baseline of concentrations in Tyrode III are indicated. Thus, the total sodium concentration after addition of 1–150 mM ranged from 146 to 295 mM. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence recorded by confocal laser scanning microscopy, correlating to [Ca 2+ ] i . (A) NaCl evokes an increase in [Ca 2+ ] i ; shown are mean and SEM. (B) Representative recording of changes in Calcium Orange® fluorescence in a single cholinergic (eGFP + ) UBC in response to increasing concentrations of NaCl (blue arrowhead) and mannitol (orange arrowhead). (C) Depicted are peak values after stimulation with mannitol (150 mM, N = 22) and NaCl (1 mM, N = 22; 10 mM, N = 22, 50 mM, N = 24, 100 mM, N = 23; 150 mM, N = 21); * P

    Journal: Frontiers in Cell and Developmental Biology

    Article Title: ENaC in Cholinergic Brush Cells

    doi: 10.3389/fcell.2018.00089

    Figure Lengend Snippet: UBC response to NaCl is ENaC dependent and not an osmolarity effect. Urethral epithelial cells of ChAT-eGFP reporter mice were isolated and UBC were identified due to eGFP fluorescence. Experiments were performed during continuous superfusion with Tyrode III buffer containing 145 mM Na + . NaCl, mannitol and amiloride were added under continuous flow of Tyrode III into the chamber, so that indicated concentrations were reached initially and then washed out. In case of NaCl, concentration changes on top of the baseline of concentrations in Tyrode III are indicated. Thus, the total sodium concentration after addition of 1–150 mM ranged from 146 to 295 mM. Y-Axis depicts arbitrary units (AU) of Calcium Orange® fluorescence recorded by confocal laser scanning microscopy, correlating to [Ca 2+ ] i . (A) NaCl evokes an increase in [Ca 2+ ] i ; shown are mean and SEM. (B) Representative recording of changes in Calcium Orange® fluorescence in a single cholinergic (eGFP + ) UBC in response to increasing concentrations of NaCl (blue arrowhead) and mannitol (orange arrowhead). (C) Depicted are peak values after stimulation with mannitol (150 mM, N = 22) and NaCl (1 mM, N = 22; 10 mM, N = 22, 50 mM, N = 24, 100 mM, N = 23; 150 mM, N = 21); * P

    Article Snippet: Test stimuli and concentrations were denatonium benzoate (25 mM; Molekula, Munich, Germany), ATP (0.5 mM; Sigma-Aldrich/Merck, Darmstadt, Germany) and NaCl (1–150 mM; Carl Roth, Karlsruhe, Germany), and inhibitors and controls included the osmolarity control mannitol (1–150 mM; Sigma-Aldrich/Merck, Darmstadt, Germany) and the ENaC inhibitor amiloride (0.01–100 μM; Sigma-Aldrich/Merck, Darmstadt, Germany).

    Techniques: Mouse Assay, Isolation, Fluorescence, Flow Cytometry, Concentration Assay, Confocal Laser Scanning Microscopy

    The latency of the first bombykal (BAL)-dependent AP was prolonged by all compounds tested. (A,B) The amiloride HMA was significantly more potent than OLC15 and MIA at both Zeitgebertimes. Box plots with whiskers from 5 to 95 percentiles. Different scales were used with interrupted error bars to improve illustration of differences. Significant differences are indicated by asterisks (exact P-values see Table 2 ; *P

    Journal: PLoS ONE

    Article Title: No Evidence for Ionotropic Pheromone Transduction in the Hawkmoth Manduca sexta

    doi: 10.1371/journal.pone.0166060

    Figure Lengend Snippet: The latency of the first bombykal (BAL)-dependent AP was prolonged by all compounds tested. (A,B) The amiloride HMA was significantly more potent than OLC15 and MIA at both Zeitgebertimes. Box plots with whiskers from 5 to 95 percentiles. Different scales were used with interrupted error bars to improve illustration of differences. Significant differences are indicated by asterisks (exact P-values see Table 2 ; *P

    Article Snippet: 5-(N-methyl-N-isobutyl)amiloride (MIA) and 5-(N,N-hexamethylene)amiloride (HMA) were obtained from Sigma-Aldrich (Sigma-Aldrich Co., Munich, Germany).

    Techniques:

    Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of dimethylamiloride (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response

    Journal: BMC Physiology

    Article Title: The gastric H,K-ATPase in stria vascularis contributes to pH regulation of cochlear endolymph but not to K secretion

    doi: 10.1186/s12899-016-0024-1

    Figure Lengend Snippet: Inhibition by amiloride analogs of strial H + flux from stria vascularis. a Summary traces of perfusion of dimethylamiloride (DMA; 30 μM; N = 5); box shows the duration of the experimental period. b summary family of curves for four Na + ,H + exchanger inhibitors: HOE694 ( N = 3–7), amiloride ( N = 4–6), S3226 ( N = 3–6) and DMA ( N = 3–9). Data from DMA and S3226 were fitted with a dual Michaelis-Menton equation consisting of one component that contributes 18 % (putative apical NHEs) and a second component that contributes 82 % (putative metabolic pathway target) of the total response

    Article Snippet: Lang), S3226 (3-[2-(3-guanidino-2-methyl-3-oxo-propenyl)-5-methy]-N-isopropylidene-2-methyl-acrylamide; gift from Sanofi-Aventis Deutschland GmbH), dimethylamiloride (DMA; Sigma A4562), ouabain (Sigma O3125), bumetanide (Sigma B3023), SCH28080 (Sigma S4443), DIDS (4,4’-Diisothiocyanatostilbene-2,2’-disulfonic acid; Sigma D3514)] and used at a final concentration of < =0.1 % DMSO or dissolved directly in bath solution [bafilomycin A1 (Sigma B1793)].

    Techniques: Inhibition

    Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), amiloride ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.

    Journal: Cephalalgia

    Article Title: Topiramate is more effective than acetazolamide at lowering intracranial pressure

    doi: 10.1177/0333102418776455

    Figure Lengend Snippet: Effect of subcutaneous administration of clinical or high dose drugs on ICP. Rats received a subcutaneous injection of topiramate ((a) and (b)), acetazolamide ((c) and (d)), amiloride ((e) and (f)), octreotide ((g) and (h)) and furosemide ((i) and (j)). ((a), (c), (e), (g), (i)) Line graphs showing the change in ICP from baseline (mmHg) ± SEM, after subcutaneous injection with either high or clinical dose of drug measured every 15 minutes for 120 minutes. ((b), (d), (f), (h), (j)) Bar graphs showing percentage of control ICP AUC over 120 minutes ± SEM, with clinical or high dose of drug. Controls: 2% NaCl for acetazolamide clinical dose, 4% Na Cl for acetazolamide high dose, and 0.9% NaCl for all other drugs tested.

    Article Snippet: Amiloride HCl (129876-100, Merck Millipore) and Octreotide acetate salt (H-5972, Bachem) were dissolved in sterile water.

    Techniques: Injection

    Fig. 1. ( A ) Sequence alignments for TM1 and TM2 regions within different members of the ENaC/Degenerin gene family. The alignment was obtained with the GCG Pileup program. Identical and similar residues are printed white on black and white on gray, respectively. Acid-sensing ion channels (ASICs), brain–liver–intestine amiloride-sensitive Na + channel (BLINaC) and epithelial Na + channel (ENaC) correspond to the rat sequences; MEC4 and UNC105 are C.elegans degenerins and dGNaC1 is a Drosophila gonad-specific amiloride-sensitive Na + channel. Note the low conservation between the sequences of TM1 when compared with TM2. ( B ) Current–voltage relationship for wild-type FaNaC expressed in HEK293 cells, in the presence of 30 µM FMRF-amide. Cells were maintained in the whole-cell configuration and voltage steps were applied from –100 to +100 mV in 20 mV increments. (Circles) Symmetrical conditions (140 mM NaCl, 0 mM KCl pH 7.4) between intracellular and extracellular medium. (Squares) Extracellular medium: 95 mM KCl, 35 mM NaCl; intracellular medium: 140 mM NaCl, 0 mM KCl. (Diamonds) Residual currents in the absence of FMRF-amide. Similar residual currents were observed in the presence of 30 µM FMRF-amide and 100 µM amiloride.

    Journal: The EMBO Journal

    Article Title: Exploration of the pore structure of a peptide-gated Na+channel

    doi: 10.1093/emboj/20.20.5595

    Figure Lengend Snippet: Fig. 1. ( A ) Sequence alignments for TM1 and TM2 regions within different members of the ENaC/Degenerin gene family. The alignment was obtained with the GCG Pileup program. Identical and similar residues are printed white on black and white on gray, respectively. Acid-sensing ion channels (ASICs), brain–liver–intestine amiloride-sensitive Na + channel (BLINaC) and epithelial Na + channel (ENaC) correspond to the rat sequences; MEC4 and UNC105 are C.elegans degenerins and dGNaC1 is a Drosophila gonad-specific amiloride-sensitive Na + channel. Note the low conservation between the sequences of TM1 when compared with TM2. ( B ) Current–voltage relationship for wild-type FaNaC expressed in HEK293 cells, in the presence of 30 µM FMRF-amide. Cells were maintained in the whole-cell configuration and voltage steps were applied from –100 to +100 mV in 20 mV increments. (Circles) Symmetrical conditions (140 mM NaCl, 0 mM KCl pH 7.4) between intracellular and extracellular medium. (Squares) Extracellular medium: 95 mM KCl, 35 mM NaCl; intracellular medium: 140 mM NaCl, 0 mM KCl. (Diamonds) Residual currents in the absence of FMRF-amide. Similar residual currents were observed in the presence of 30 µM FMRF-amide and 100 µM amiloride.

    Article Snippet: FMRF-amide (Sigma), amiloride (Sigma), MTSET (Toronto Research Chemicals) and MTSES (Toronto Research Chemicals) were diluted in the 140 mM NaCl solution to final concentrations of 30 µM, 100 µM, 1 mM and 10 mM, respectively.

    Techniques: Sequencing

    Fig. 2. Susceptibility of wild-type FaNaC and several mutants to 1 mM MTSET. Positive transfectants in the whole-cell clamp configuration were maintained at –40 mV. Bath and pipette solutions were of identical composition (140 mM NaCl, 10 mM HEPES, 1 mM CaCl 2 pH 7.4). Cells were briefly perfused twice with the same solution containing 30 µM FMRF-amide to determine the maximal amplitude of the stimulation ( I max ) before being rinsed until the baseline current was reached, and then perfused with 1 mM MTSET in the same solution for 3 min. The third peak corresponds to the channel stimulation by 30 µM FMRF-amide after 3 min of MTSET application followed by 1 min of rinse. In these conditions, the wild-type channel (WT) and mutants such as Ile75Cys are not affected by 1 mM MTSET, while mutants such as Val74Cys show a marked inhibition. Note that the Ser86Cys mutant, which exhibits a strong desensitization upon FMRF-amide repeated stimulations, regains activity and does not desensitize following 3 min of 1 mM MTSET exposure. F, 30 µM FMRF-amide; R, rinse.

    Journal: The EMBO Journal

    Article Title: Exploration of the pore structure of a peptide-gated Na+channel

    doi: 10.1093/emboj/20.20.5595

    Figure Lengend Snippet: Fig. 2. Susceptibility of wild-type FaNaC and several mutants to 1 mM MTSET. Positive transfectants in the whole-cell clamp configuration were maintained at –40 mV. Bath and pipette solutions were of identical composition (140 mM NaCl, 10 mM HEPES, 1 mM CaCl 2 pH 7.4). Cells were briefly perfused twice with the same solution containing 30 µM FMRF-amide to determine the maximal amplitude of the stimulation ( I max ) before being rinsed until the baseline current was reached, and then perfused with 1 mM MTSET in the same solution for 3 min. The third peak corresponds to the channel stimulation by 30 µM FMRF-amide after 3 min of MTSET application followed by 1 min of rinse. In these conditions, the wild-type channel (WT) and mutants such as Ile75Cys are not affected by 1 mM MTSET, while mutants such as Val74Cys show a marked inhibition. Note that the Ser86Cys mutant, which exhibits a strong desensitization upon FMRF-amide repeated stimulations, regains activity and does not desensitize following 3 min of 1 mM MTSET exposure. F, 30 µM FMRF-amide; R, rinse.

    Article Snippet: FMRF-amide (Sigma), amiloride (Sigma), MTSET (Toronto Research Chemicals) and MTSES (Toronto Research Chemicals) were diluted in the 140 mM NaCl solution to final concentrations of 30 µM, 100 µM, 1 mM and 10 mM, respectively.

    Techniques: Transferring, Inhibition, Mutagenesis, Activity Assay