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  • 99
    Millipore nifedipine
    Calcium influx into RBC upon sensitization with anti-9- O -AcSGP IgG VL antibodies. For all experiments, erythrocytes were washed and loaded with the Ca 2+ -indicator fluorochrome Fluo-3/AM in Ringer solution. A23187-treated Fluo-3/AM-loaded RBC VL served as positive controls (black line) and A23187-treated cells in the presence of EGTA as negative controls (gray background). A. Fluo-3/AM loaded RBC VL and RBC N (2×10 7 ) were left unsensitized or sensitized with varying concentrations of anti-9- O -AcSGP IgG VL or anti-9- O -AcSGP IgG NHS , respectively, (0–40 µg/ml) in Ringer solution for 30 min at 37°C. After washing the cells twice with same solution the fluorescence intensities of the Ca 2+ . B. Cells treated as in A with 0.5 µg/ml (red line), 1.0 µg/ml (green line) and 2.5 µg/ml (pink line) anti-9- O -AcSGP IgG VL were analyzed by flow cytometry to determine the fraction of responding RBC. C. Similarly, RBC N were analyzed without and with sensitization with 5 µg/ml anti-9-O-AcSGP IgG NHS (blue line). D–E. Time dependent increase of intracellular Ca 2+ in RBC VL after sensitization with anti-9- O -AcSGP IgG VL (2.5 µg/ml) for 5 min (red line), 10 min (green line), 20 min (pink line) and 30 min (blue line) at 37°C. Cells were washed and analyzed by flow cytometry. D. Histogram representation; E. Mean fluorescence intensities calculated from D. F–G. Inhibition of the Ca 2+ influx into RBC sensitized with anti-9- O -AcSGP IgG VL . Prior to sensitization with anti-9- O -AcSGP-IgG VL (2.5 µg/ml), the cells were incubated without (green line) or with the P/Q-type channel blocker ω-agatoxin TK (1 µM, pink line, F ), or without (green line) or with the L-type channel blocker <t>nifedipine</t> (10 µM, blue line, G ) and analyzed by flow cytometry.
    Nifedipine, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 2719 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Tocris nifedipine
    Spontaneous neuronal calcium activity in GC-P and PGN-P in the RMS, RMS-OB and deep GCL. A. Schematic representation of an olfactory bulb sagittal slice. B. Ai14/GCaMP6s P0 animals received lateral or medial electroporation of a Cre mRNAs. C. Calcium activity was recorded on acute brain slices between 6 and 10 dpe. Red and green cells represent medial and lateral cells migrating in the RMS, RMS-OB or in the deep GCL. D.F.H. Raster plot of calcium activity of GC-P (left) and PGN-P (right) recorded in the RMS (D), RMS OB (F) and deep GCL (H). E.G.I. Percentage of active cells, frequency and mean amplitude for GC-P and PGN-P in the RMS (E), RMS-OB (G) and in the deep GCL (I). Cells were defined as active if they showed at least one detectable calcium transient. The calcium activity of GC-P greatly increased upon arrival in the RMS-OB and the deep GCL, in contrast with PGN-P activity. N = 6 slices per condition. Error bars are SEM. J. Left: Raster plot of calcium activity of GC-P in the deep GCL in control and calcium-free medium (No Calcium + 2mM EGTA). Right: The percentage of active cells is highly decreased in calcium free medium. K. Left: Raster plot of calcium activity of GC-P in the deep GCL in control condition and <t>nifedipine</t> (10 μM) supplemented medium. Right: The percentage of active after incubation with the L-Type Voltage Gated Calcium Channels antagonist Nifedipine at 10 μM is highly decreased.
    Nifedipine, supplied by Tocris, used in various techniques. Bioz Stars score: 99/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Neuroscience Information Framework nifedipine
    Intracellular Ca 2+ signaling was reduced by L-803,087 in ganglion cells in retinal flat mount. A : applications of 52 mM K + solution for 30 s depolarized ganglion cells and produced transient increases in intracellular Ca 2+ concentration, measured with the dye fura-2. Traces ( left ) show that, in the absence of drug, the second of paired K + applications produced a smaller peak Ca 2+ signal. On the right , the paired K + pulses, with L-803,087 (100 nM) applied 30 s prior to the second K + application, show greater reduction of the Ca 2+ signal compared with control. Far right panel shows a fluorescent image (stimulated at 380 nm) of a fura-2-containing ganglion cell in the flat-mount retina. B : mean data show amplitudes of the second high K + response expressed as a percentage of first peak in control, L-803,087 (L-803, 100 nM), <t>nifedipine</t> (NIF, 10 μM), and nifedipine plus L-803,087. The mean Ca 2+ transient amplitudes of all 3 drug treatment groups (L-803, NIF, L-803 + NIF) were significantly smaller than those of the control group, but they did not differ among themselves ( n = 21–89 cells/group; * P
    Nifedipine, supplied by Neuroscience Information Framework, used in various techniques. Bioz Stars score: 93/100, based on 249 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Alomone Labs nifedipine
    Effect of ion channel blockers on L55P TTR-induced calcium influx and analysis of sensitivity to icilin and capsaicin , (A) L55P was applied to DRG growth cones in the presence of VGCC inhibitors <t>(nifedipine,</t> ω-agatoxin IVA, ω-conotoxin GIVA), Na V inhibitors (tetrodotoxin, ambroxol and carbamazepine) and TRP inhibitors (SKF-96365, BCTC). The resulting maximal calcium influx (max ΔF/F 0 ) calculated over the imaging period (7 min) was calculated. (B) Effect of capsaicin (1 μM) and icilin (100 μM) on cytosolic calcium in DRG growth cones in culture. When DRG cultures were pre-treated with ambroxol (5 μM), icilin-induced calcium fluorescence was significantly decreased. All graphs show maximal ΔF/F 0 ± SEM for n = 12-24 growth cones. Significant differences from control values are depicted as: * p
    Nifedipine, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 92/100, based on 53 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    FUJIFILM nifedipine
    Voltage-dependent Ba 2+ currents in the epithelioid cells Epithelioid cells were voltage clamped at -70 mV in the presence of tetrodotoxin (0.2 μM) and BaCl 2 (5 mM) instead of CaCl 2 . A , representative recordings of current responses of an epithelioid cell to depolarizing potentials for 30 ms between -60 and +60 mV in 10 mV increments before (Control) and during exposure to CoCl 2 (2 mM), and after its removal (Washout). B , peak inward current-voltage relationships were obtained from the cell before (▪) and during (•) exposure to CoCl 2 , and after its removal (▴). C , effects of <t>nifedipine</t> and ω-conotoxin GVIA on peak Ba 2+ currents in an epithelioid cell. At -70 mV in the presence of tetrodotoxin (0.2 μM), TEA-Cl (5 mM) and BaCl 2 (5 mM), Ba 2+ currents were evoked by depolarizing steps for 15 ms to +10 mV with 20 s intervals. The peak Ba 2+ currents are expressed as percentages of the first Ba 2+ current recorded in the absence of blockers and plotted against time. Nifedipine (1 μM) and ω-conotoxin GVIA (1 μM) were applied during the periods indicated by horizontal lines starting with an arrow. Inset, superimposed current traces with letters corresponding to those in the peak current trace. The dotted line indicates the zero current level.
    Nifedipine, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam nifedipine
    Effect of extracellular Ca 2+ removal, <t>nifedipine,</t> CPA, and ryanodine on PMA-induced airway SMC twitching. Line scans from airway wall regions obtained from phase-contrast images (similar to that presented in Fig. 1 ) showing airway SMC twitching induced by 10 µM PMA (added 30 min before the recordings started) and their sensitivity to: (A) Ca 2+ -free sHBSS, (B) 10 µM nifedipine, (C) 10 µM CPA, or (D) 25 µM ryanodine. PMA-induced SMC twitching was inhibited by removal of extracellular Ca 2+ , nifedipine, CPA, and ryanodine. Each line scan is representative of three experiments in lung slices from two mice.
    Nifedipine, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bayer AG nifedipine
    Displacement by <t>nifedipine</t> of [ 3 H]-R-PIA and [ 3 H]-XAC bound to membrane receptors. Open symbols [ 3 H]-R-PIA binding (K i app. 4.15 ± 1.37 × 10 −6 , B max app. 653 fmol/mg protein); closed symbols [ 3 H]-XAC (K i app. 1.86 ± 0.42 × 10 −6 M, B max app. 868 fmol/mg protein).
    Nifedipine, supplied by Bayer AG, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Biomol GmbH nifedipine
    High voltage activation calcium channels' L-type regulation and pathway signaling Ca 2+ /CaM-K II affect GH release in bovine somatotropes. (a) Bar graph illustrating the regulation of hormone release by GHRP-6 (100 nM) applied alone or in the presence of <t>nifedipine</t> (0.5 μ M) a blocker of HVACC L-type. (b) Average amount of GH released in the absences (control) and after treatment with the signaling pathway Ca 2+ /CaM-K II inhibitor KN-62 (10 μ M) alone or in combination with GHRP-6 (100 nM). Each value represents the mean ± SE of determinations performed in triplicate from three independent experiments. The asterisks denote significant differences ( P
    Nifedipine, supplied by Biomol GmbH, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Nacalai nifedipine
    Profiles of Relaxation by VDCC Blockers in Both WKY and SHR. Representative traces of <t>nifedipine</t> (A)-, verapamil (C)-, and Trp-His (E)-induced relaxation in PE (1 µM)-contracted aortic rings from both 8- and 40-week WKY and SHR. The relaxation activity was represented as EC 50 values for nifedipine (B), verapamil (D), and Trp-His (F). Results are expressed as the mean ± SEM values (n = 3–7). * P
    Nifedipine, supplied by Nacalai, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Bayer AG oral nifedipine
    Profiles of Relaxation by VDCC Blockers in Both WKY and SHR. Representative traces of <t>nifedipine</t> (A)-, verapamil (C)-, and Trp-His (E)-induced relaxation in PE (1 µM)-contracted aortic rings from both 8- and 40-week WKY and SHR. The relaxation activity was represented as EC 50 values for nifedipine (B), verapamil (D), and Trp-His (F). Results are expressed as the mean ± SEM values (n = 3–7). * P
    Oral Nifedipine, supplied by Bayer AG, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Tokyo Chemical Industry nifedipine
    Time-course analysis of drug absorption into PDMS, FEPM, and polystyrene (PS) culture plates (control). Drug concentrations after 0.5, 1, 2, 3, 6, and 24-h incubation of <t>nifedipine,</t> Bay K8644, and coumarin evaluated by high-performance liquid chromatography (HPLC, n = 3, * p
    Nifedipine, supplied by Tokyo Chemical Industry, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Mylan nifedipine gits
    Curves for ABPM data plotted using the local regression LOESS function in R statistical software. Readings from 64 timepoints over 24 h for systolic and diastolic blood pressure (SBP and DBP) and heart rate (HR) provided 2,560 values for each parameter from 20 patients over two study periods. Solid‐red lines represent data fit from <t>Mylan‐Nifedipine‐XL.</t> Dashed‐green lines represent data fit from <t>Nifedipine‐GITS.</t> SBP curves for two patients are statistically different and continue to further separate during the terminal 8 h (nocturnal period) of the dosing interval, as delivery from Mylan‐Nifedipine‐XL is predicted to decline in a first‐order pattern. DBP curves follow a similar pattern, but are less affected by action of dihydropyridines and were not statistically different. HR remains clinically unaffected by constant, low‐rate delivery of nifedipine, varying by only 15 bpm over the course of the day.
    Nifedipine Gits, supplied by Mylan, 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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    94
    Selleck Chemicals nifedipine
    Neuritic degeneration induced by polybrene is dependent on Ca 2+ influx. a , b Ca 2+ influx induced by polybrene was inhibited by EGTA or <t>nifedipine.</t> Neurons were labeled by Fluo-4. a The time course of relative Fluo-4 fluorescence intensity was recorded. b The relative Fluo-4 fluorescence intensity at 3000 s. The fluorescence intensity of Fluo-4 at initiating time was normalized to 100 ( n ≥ 6, *** P
    Nifedipine, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 94/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher nifedipine
    mAChR agonist carbachol does not affect the depolarization-induced whole cell steady-state K Ca 1.1 currents under conditions of pharmacological inhibition of the major cellular sources of Ca 2+ for K Ca 1.1 channel activation in freshly-isolated human DSM cells A) Representative original recordings illustrate the depolarization-induced whole cell steady-state K Ca 1.1 currents in the absence (control) and in the presence of carbachol (1 μM). B) The current-voltage relationship curve summarizes the lack of carbachol effects on the whole cell steady-state K Ca 1.1 currents (n=9, N=6; P > 0.05). Steady-state K Ca 1.1 currents were recorded in the presence of ryanodine (30 μM), thapsigargin (100 nM), and <t>nifedipine</t> (1 μM). The bath solution contained 134 mM NaCl, 6 mM KCl, and 2 mM CaCl 2 , and the pipette 110 mM potassium aspartate, 30 mM KCl, 10 mM NaCl, and
    Nifedipine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Valiant nifedipine
    Cumulative data showing the relative frequencies of LCR events during the peak of the responses to hyperosmotic sucrose, hyperosmotic CaCl 2 , or after transient exposure to a hypoosmotic solution in the absence or presence of <t>nifedipine.</t> ***, statistically significant differences (P
    Nifedipine, supplied by Valiant, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Simcyp nifedipine
    Predicted and observed (A) plasma concentration profile and (B) change in systolic blood pressure after a single dose of <t>nifedipine</t> 60 mg GITS in North European hypertensive subjects ( Meredith and Elliott, 2004 ). Predicted and observed (C) plasma concentration profile and (D) change in systolic blood pressure after the initial dose and daily dosing of nifedipine 30 mg GITS for 15 days in North European hypertensive subjects ( Brown and Toal, 2008 ).
    Nifedipine, supplied by Simcyp, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Bayer AG m nifedipine
    Representative traces of single N-channel activity recorded from cell-attached patches of control NG108-15 and cav1(+) cells. ( A ) Traces from a control NG108-15 cell obtained with pulses from −40 to +20 mV of 180 ms each. The averaged current at the bottom (−0.051 pA) was calculated over 257 sweeps including nulls. ( B ) Traces from a cav1(+) cell. The averaged current at the bottom is 69% smaller (−0.016 pA) than control (423 sweeps including nulls). The patch pipette contained 100 mM Ba 2+ and the L-type Ca 2+ channel blocker <t>nifedipine</t> (10 μ M).
    M Nifedipine, supplied by Bayer AG, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Calcium influx into RBC upon sensitization with anti-9- O -AcSGP IgG VL antibodies. For all experiments, erythrocytes were washed and loaded with the Ca 2+ -indicator fluorochrome Fluo-3/AM in Ringer solution. A23187-treated Fluo-3/AM-loaded RBC VL served as positive controls (black line) and A23187-treated cells in the presence of EGTA as negative controls (gray background). A. Fluo-3/AM loaded RBC VL and RBC N (2×10 7 ) were left unsensitized or sensitized with varying concentrations of anti-9- O -AcSGP IgG VL or anti-9- O -AcSGP IgG NHS , respectively, (0–40 µg/ml) in Ringer solution for 30 min at 37°C. After washing the cells twice with same solution the fluorescence intensities of the Ca 2+ . B. Cells treated as in A with 0.5 µg/ml (red line), 1.0 µg/ml (green line) and 2.5 µg/ml (pink line) anti-9- O -AcSGP IgG VL were analyzed by flow cytometry to determine the fraction of responding RBC. C. Similarly, RBC N were analyzed without and with sensitization with 5 µg/ml anti-9-O-AcSGP IgG NHS (blue line). D–E. Time dependent increase of intracellular Ca 2+ in RBC VL after sensitization with anti-9- O -AcSGP IgG VL (2.5 µg/ml) for 5 min (red line), 10 min (green line), 20 min (pink line) and 30 min (blue line) at 37°C. Cells were washed and analyzed by flow cytometry. D. Histogram representation; E. Mean fluorescence intensities calculated from D. F–G. Inhibition of the Ca 2+ influx into RBC sensitized with anti-9- O -AcSGP IgG VL . Prior to sensitization with anti-9- O -AcSGP-IgG VL (2.5 µg/ml), the cells were incubated without (green line) or with the P/Q-type channel blocker ω-agatoxin TK (1 µM, pink line, F ), or without (green line) or with the L-type channel blocker nifedipine (10 µM, blue line, G ) and analyzed by flow cytometry.

    Journal: PLoS ONE

    Article Title: Sialoglycosylation of RBC in Visceral Leishmaniasis Leads to Enhanced Oxidative Stress, Calpain-Induced Fragmentation of Spectrin and Hemolysis

    doi: 10.1371/journal.pone.0042361

    Figure Lengend Snippet: Calcium influx into RBC upon sensitization with anti-9- O -AcSGP IgG VL antibodies. For all experiments, erythrocytes were washed and loaded with the Ca 2+ -indicator fluorochrome Fluo-3/AM in Ringer solution. A23187-treated Fluo-3/AM-loaded RBC VL served as positive controls (black line) and A23187-treated cells in the presence of EGTA as negative controls (gray background). A. Fluo-3/AM loaded RBC VL and RBC N (2×10 7 ) were left unsensitized or sensitized with varying concentrations of anti-9- O -AcSGP IgG VL or anti-9- O -AcSGP IgG NHS , respectively, (0–40 µg/ml) in Ringer solution for 30 min at 37°C. After washing the cells twice with same solution the fluorescence intensities of the Ca 2+ . B. Cells treated as in A with 0.5 µg/ml (red line), 1.0 µg/ml (green line) and 2.5 µg/ml (pink line) anti-9- O -AcSGP IgG VL were analyzed by flow cytometry to determine the fraction of responding RBC. C. Similarly, RBC N were analyzed without and with sensitization with 5 µg/ml anti-9-O-AcSGP IgG NHS (blue line). D–E. Time dependent increase of intracellular Ca 2+ in RBC VL after sensitization with anti-9- O -AcSGP IgG VL (2.5 µg/ml) for 5 min (red line), 10 min (green line), 20 min (pink line) and 30 min (blue line) at 37°C. Cells were washed and analyzed by flow cytometry. D. Histogram representation; E. Mean fluorescence intensities calculated from D. F–G. Inhibition of the Ca 2+ influx into RBC sensitized with anti-9- O -AcSGP IgG VL . Prior to sensitization with anti-9- O -AcSGP-IgG VL (2.5 µg/ml), the cells were incubated without (green line) or with the P/Q-type channel blocker ω-agatoxin TK (1 µM, pink line, F ), or without (green line) or with the L-type channel blocker nifedipine (10 µM, blue line, G ) and analyzed by flow cytometry.

    Article Snippet: To determine the Ca2+ channel type, Fluo 3-loaded RBCVL (2×107 ) in Ringer solution containing Ca2+ (5 mM) were preincubated without or with the indicated doses of ω-agatoxin TK, a spider venom peptide (P/Q-type Ca2+ channel blocker, Calbiochem) or nifedipine (L-type Ca2+ channel blocker, Sigma) for 20 min at 37°C, washed, sensitized with anti-9- O -AcSGP IgG (2.5 µg/ml) as above and analyzed by flow cytometry .

    Techniques: Fluorescence, Flow Cytometry, Cytometry, Inhibition, Incubation

    The DHPR is the membrane potential sensor for the slow calcium signal. Myotubes were stimulated with 400 pulses of 1 ms at 45 Hz. ( A ) The myotubes in nominally zero calcium ( open circles ) were exposed to 1 μ M of (−)-S-Bay K 8644, a DHPR activator ( open squares ), and 1 μ M nifedipine ( open triangles ). The mean for each condition of at least 13 independent experiments ± SE is shown. ( B ) Myotubes derived from the dysgenic skeletal muscle cell line (GLT, open squares ), the wild-type mice derived cell line (NLT, open circles ), and a GLT clone stable-transfected with the DHPR- α 1 S subunit (GLT- α 1, open triangles ) were exposed to the same stimulation protocol as described in A . The experiments were performed in the absence of extracellular calcium. The mean of at least six independent experiments ± SE for each condition is shown. The lower trace indicates the time of stimulation.

    Journal: Biophysical Journal

    Article Title: Slow Calcium Signals after Tetanic Electrical Stimulation in Skeletal Myotubes

    doi:

    Figure Lengend Snippet: The DHPR is the membrane potential sensor for the slow calcium signal. Myotubes were stimulated with 400 pulses of 1 ms at 45 Hz. ( A ) The myotubes in nominally zero calcium ( open circles ) were exposed to 1 μ M of (−)-S-Bay K 8644, a DHPR activator ( open squares ), and 1 μ M nifedipine ( open triangles ). The mean for each condition of at least 13 independent experiments ± SE is shown. ( B ) Myotubes derived from the dysgenic skeletal muscle cell line (GLT, open squares ), the wild-type mice derived cell line (NLT, open circles ), and a GLT clone stable-transfected with the DHPR- α 1 S subunit (GLT- α 1, open triangles ) were exposed to the same stimulation protocol as described in A . The experiments were performed in the absence of extracellular calcium. The mean of at least six independent experiments ± SE for each condition is shown. The lower trace indicates the time of stimulation.

    Article Snippet: Before a specific experiment, the cells were incubated from 30 to 60 min with 10 μ M tetrodotoxin (TTX) (Sigma-Aldrich), 10–30 μ M ryanodine (Sigma-Aldrich), 1 μ M nifedipine (Sigma-Aldrich), 2 μ M (−)S-Bay K 8644 (RBI, Natick, MA), 5 μ M xestospongin C (Calbiochem, La Jolla, CA), or 30 μ M (Sigma-Aldrich), in Krebs buffer (145 mM NaCl, 5 mM KCl, 1 mM CaCl2 , 1 mM MgCl2 , 10 mM HEPES-Na, 5.6 mM glucose, pH 7.4).

    Techniques: Mass Spectrometry, Derivative Assay, Mouse Assay, Transfection

    Blocking of voltage-gated ion channels using a high dose of nifedipine (Nf) during dynamic culture had noticeable effects on joint shapes compared with dynamic vehicle controls, with the knee joints of the high-dose group being more similar to the static groups. Administration of the drug during static cultures had no obvious effects on shape. Low dose, 0.1 mM; high dose, 1 mM; squares, dominance of the lateral condyle over the medial condyle; hollow arrows, posterior curl of condyles seen in moved explants; *, dorsal groove between the two femoral condyles; single-edged bars, intercondylar fossa width; +, concave curve of the tibial plateau; filled arrows, anterior tibial crest.

    Journal: Philosophical Transactions of the Royal Society B: Biological Sciences

    Article Title: Blocking mechanosensitive ion channels eliminates the effects of applied mechanical loading on chick joint morphogenesis

    doi: 10.1098/rstb.2017.0317

    Figure Lengend Snippet: Blocking of voltage-gated ion channels using a high dose of nifedipine (Nf) during dynamic culture had noticeable effects on joint shapes compared with dynamic vehicle controls, with the knee joints of the high-dose group being more similar to the static groups. Administration of the drug during static cultures had no obvious effects on shape. Low dose, 0.1 mM; high dose, 1 mM; squares, dominance of the lateral condyle over the medial condyle; hollow arrows, posterior curl of condyles seen in moved explants; *, dorsal groove between the two femoral condyles; single-edged bars, intercondylar fossa width; +, concave curve of the tibial plateau; filled arrows, anterior tibial crest.

    Article Snippet: Voltage-gated calcium ion channels were blocked using nifedipine (Nf) dissolved in DMSO (Sigma Aldrich, USA).

    Techniques: Blocking Assay

    Blocking of voltage-gated ion channels using a high dose of nifedipine (Nf) during dynamic cultures led to significant reductions in 8 out of 11 measurements performed. No measurements were significantly different between the dynamic high-dose group and any of the static groups. Circle (for dynamic cultures) and triangles (for static cultures) represent individual data points. Black horizontal lines represent significant differences between groups (significance for p -value less than 0.05). Full details of statistical results, including means, standard deviations (s.d.) and p -values, are reported in the electronic supplementary material, tables S1–S10.

    Journal: Philosophical Transactions of the Royal Society B: Biological Sciences

    Article Title: Blocking mechanosensitive ion channels eliminates the effects of applied mechanical loading on chick joint morphogenesis

    doi: 10.1098/rstb.2017.0317

    Figure Lengend Snippet: Blocking of voltage-gated ion channels using a high dose of nifedipine (Nf) during dynamic cultures led to significant reductions in 8 out of 11 measurements performed. No measurements were significantly different between the dynamic high-dose group and any of the static groups. Circle (for dynamic cultures) and triangles (for static cultures) represent individual data points. Black horizontal lines represent significant differences between groups (significance for p -value less than 0.05). Full details of statistical results, including means, standard deviations (s.d.) and p -values, are reported in the electronic supplementary material, tables S1–S10.

    Article Snippet: Voltage-gated calcium ion channels were blocked using nifedipine (Nf) dissolved in DMSO (Sigma Aldrich, USA).

    Techniques: Blocking Assay

    CEPN inhibits ACh-induced precontraction. (a) ACh induced a contraction in a tracheal ring that was inhibited completely by CEPN. This experiment was repeated in 6 rings. (b) Nifedipine partially reduced the ACh-induced contraction in a dose-dependent manner. The resistant contraction was inhibited by CEPN. The summary results from 6 experiments are shown in (c). (d) In the presence of nifedipine, ACh induced a typical contraction, which was dose-dependently inhibited by CEPN. The summary results from 6 experiments exhibited in (e). These results show that CEPN inhibits VDCCs and another pathway to induce relaxation.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Relaxant Action of Plumula Nelumbinis Extract on Mouse Airway Smooth Muscle

    doi: 10.1155/2015/523640

    Figure Lengend Snippet: CEPN inhibits ACh-induced precontraction. (a) ACh induced a contraction in a tracheal ring that was inhibited completely by CEPN. This experiment was repeated in 6 rings. (b) Nifedipine partially reduced the ACh-induced contraction in a dose-dependent manner. The resistant contraction was inhibited by CEPN. The summary results from 6 experiments are shown in (c). (d) In the presence of nifedipine, ACh induced a typical contraction, which was dose-dependently inhibited by CEPN. The summary results from 6 experiments exhibited in (e). These results show that CEPN inhibits VDCCs and another pathway to induce relaxation.

    Article Snippet: Reagents Nifedipine, acetylcholine chloride (ACh), niflumic acid (NA), tetraethylammonium chloride (TEA), and pyrazole 3 (Pyr3) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques:

    Pyr3 inhibits ACh-induced contraction. (a) Pyr3, a blocker of TRPC3 and Orai1 channels, partially inhibited ACh-induced contraction in a dose-dependent manner. The remained contraction was completely blocked by 1 mg/mL CEPN. The dose-response from 4 experiments is shown in (b). (c) Comparison of the relaxant effects of CEPN, nifedipine, and Pyr3 on ACh-induced contraction. ** P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Relaxant Action of Plumula Nelumbinis Extract on Mouse Airway Smooth Muscle

    doi: 10.1155/2015/523640

    Figure Lengend Snippet: Pyr3 inhibits ACh-induced contraction. (a) Pyr3, a blocker of TRPC3 and Orai1 channels, partially inhibited ACh-induced contraction in a dose-dependent manner. The remained contraction was completely blocked by 1 mg/mL CEPN. The dose-response from 4 experiments is shown in (b). (c) Comparison of the relaxant effects of CEPN, nifedipine, and Pyr3 on ACh-induced contraction. ** P

    Article Snippet: Reagents Nifedipine, acetylcholine chloride (ACh), niflumic acid (NA), tetraethylammonium chloride (TEA), and pyrazole 3 (Pyr3) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques:

    CEPN blocks VDCC currents. (a) The protocol used to measure VDCC currents in single ASMCs. (b) VDCC currents, recorded following depolarizations, were blocked by CEPN and nifedipine, respectively. (c) I - V relationships constructed based on the results of 5 to 6 experiments. These data indicate that CEPN blocks VDCCs.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Relaxant Action of Plumula Nelumbinis Extract on Mouse Airway Smooth Muscle

    doi: 10.1155/2015/523640

    Figure Lengend Snippet: CEPN blocks VDCC currents. (a) The protocol used to measure VDCC currents in single ASMCs. (b) VDCC currents, recorded following depolarizations, were blocked by CEPN and nifedipine, respectively. (c) I - V relationships constructed based on the results of 5 to 6 experiments. These data indicate that CEPN blocks VDCCs.

    Article Snippet: Reagents Nifedipine, acetylcholine chloride (ACh), niflumic acid (NA), tetraethylammonium chloride (TEA), and pyrazole 3 (Pyr3) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: Construct

    Relaxant effects of CEPN on high K + -induced precontraction. (a) High K + induced a steady-state contraction in a mouse tracheal ring, which was inhibited by CEPN in a concentration-dependent manner. (b) Dose-relaxation curve of CEPN based on the results of 7 different experiments shown in (a). (c) High K + -induced precontraction was completely blocked by nifedipine. This experiment was performed in 8 tracheal rings from 8 mice, and the result was reproducible across experiments. (d) CEPN had no effect on resting tension in 4 rings. These results indicate that the CEPN-induced relaxation might result from blockade of VDCCs.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Relaxant Action of Plumula Nelumbinis Extract on Mouse Airway Smooth Muscle

    doi: 10.1155/2015/523640

    Figure Lengend Snippet: Relaxant effects of CEPN on high K + -induced precontraction. (a) High K + induced a steady-state contraction in a mouse tracheal ring, which was inhibited by CEPN in a concentration-dependent manner. (b) Dose-relaxation curve of CEPN based on the results of 7 different experiments shown in (a). (c) High K + -induced precontraction was completely blocked by nifedipine. This experiment was performed in 8 tracheal rings from 8 mice, and the result was reproducible across experiments. (d) CEPN had no effect on resting tension in 4 rings. These results indicate that the CEPN-induced relaxation might result from blockade of VDCCs.

    Article Snippet: Reagents Nifedipine, acetylcholine chloride (ACh), niflumic acid (NA), tetraethylammonium chloride (TEA), and pyrazole 3 (Pyr3) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques: Concentration Assay, Mouse Assay

    CEPN blocks ACh-evoked Ca 2+ influx. Representative result of 4 experiments in the presence of nifedipine. Under Ca 2+ -free conditions (0 Ca 2+ + 0.5 mM EGTA), ACh induced a fast transient contraction. Following the addition of 2 mM Ca 2+ , a strong sustained contraction occurred, which was entirely reversed by CEPN. These experiments indicate that CEPN inhibits the nifedipine-resistant Ca 2+ influx activated by ACh.

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Relaxant Action of Plumula Nelumbinis Extract on Mouse Airway Smooth Muscle

    doi: 10.1155/2015/523640

    Figure Lengend Snippet: CEPN blocks ACh-evoked Ca 2+ influx. Representative result of 4 experiments in the presence of nifedipine. Under Ca 2+ -free conditions (0 Ca 2+ + 0.5 mM EGTA), ACh induced a fast transient contraction. Following the addition of 2 mM Ca 2+ , a strong sustained contraction occurred, which was entirely reversed by CEPN. These experiments indicate that CEPN inhibits the nifedipine-resistant Ca 2+ influx activated by ACh.

    Article Snippet: Reagents Nifedipine, acetylcholine chloride (ACh), niflumic acid (NA), tetraethylammonium chloride (TEA), and pyrazole 3 (Pyr3) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

    Techniques:

    Effect of nifedipine on diameter and relative intracellular calcium level changes . Average data of diameter changes (A) and intracellular calcium level as indicated by R340/380 (B) of small mesenteric arteries isolated from young and old rats that were pre-contracted with 60 mM KCl and then relaxed with 1 μM nifedipine. Values are mean ± SEM of % changes from KCl contraction. * denotes significance at P ≤ 0.05.

    Journal: Frontiers in Physiology

    Article Title: Aging Reduces L-Type Calcium Channel Current and the Vasodilatory Response of Small Mesenteric Arteries to Calcium Channel Blockers

    doi: 10.3389/fphys.2016.00171

    Figure Lengend Snippet: Effect of nifedipine on diameter and relative intracellular calcium level changes . Average data of diameter changes (A) and intracellular calcium level as indicated by R340/380 (B) of small mesenteric arteries isolated from young and old rats that were pre-contracted with 60 mM KCl and then relaxed with 1 μM nifedipine. Values are mean ± SEM of % changes from KCl contraction. * denotes significance at P ≤ 0.05.

    Article Snippet: Chemicals Nifedipine, PE, SNP, and acetylcholine (ACh) were obtained from Sigma (Germany).

    Techniques: Isolation

    L-type voltage-gated calcium channel currents . (A) Representative traces of inward Ba 2+ currents generated by 8 mV steps from a holding potential of −70 to +58 mV in vascular smooth muscle cells (VSMCs) isolated from mesenteric arteries of young and old rats. Membrane capacitances were 32 and 48 pF, respectively. Currents from both animal groups (cont) were equally blocked by 1 μM nifedipine (nif). (B) Averaged current-voltage (I-V) relationships showing current density in VSMCs of arteries from young compared to old. Steady state activation (C) , and inactivation (D) curves are represented as I/I max and G/G max ; respectively.

    Journal: Frontiers in Physiology

    Article Title: Aging Reduces L-Type Calcium Channel Current and the Vasodilatory Response of Small Mesenteric Arteries to Calcium Channel Blockers

    doi: 10.3389/fphys.2016.00171

    Figure Lengend Snippet: L-type voltage-gated calcium channel currents . (A) Representative traces of inward Ba 2+ currents generated by 8 mV steps from a holding potential of −70 to +58 mV in vascular smooth muscle cells (VSMCs) isolated from mesenteric arteries of young and old rats. Membrane capacitances were 32 and 48 pF, respectively. Currents from both animal groups (cont) were equally blocked by 1 μM nifedipine (nif). (B) Averaged current-voltage (I-V) relationships showing current density in VSMCs of arteries from young compared to old. Steady state activation (C) , and inactivation (D) curves are represented as I/I max and G/G max ; respectively.

    Article Snippet: Chemicals Nifedipine, PE, SNP, and acetylcholine (ACh) were obtained from Sigma (Germany).

    Techniques: Generated, Isolation, Activation Assay

    Effect of calcium channel blockers on KCl-induced contraction . Responses of mesenteric arteries isolated from young and old rats that were pre-contracted with 100 mM KCl and relaxed with cumulative concentrations of three calcium channel blockers, nifedipine (A) , verapamil (B) , and diltiazem (C) . (D) Shows the relationship between cumulative concentrations of sodium nitroprusside (SNP) and % relaxation of phenylephrine (PE; 4 μM)-induced contraction.

    Journal: Frontiers in Physiology

    Article Title: Aging Reduces L-Type Calcium Channel Current and the Vasodilatory Response of Small Mesenteric Arteries to Calcium Channel Blockers

    doi: 10.3389/fphys.2016.00171

    Figure Lengend Snippet: Effect of calcium channel blockers on KCl-induced contraction . Responses of mesenteric arteries isolated from young and old rats that were pre-contracted with 100 mM KCl and relaxed with cumulative concentrations of three calcium channel blockers, nifedipine (A) , verapamil (B) , and diltiazem (C) . (D) Shows the relationship between cumulative concentrations of sodium nitroprusside (SNP) and % relaxation of phenylephrine (PE; 4 μM)-induced contraction.

    Article Snippet: Chemicals Nifedipine, PE, SNP, and acetylcholine (ACh) were obtained from Sigma (Germany).

    Techniques: Isolation

    Experimental comparison of OptoHTS versus ‘spark'-OptoHTS for functional drug testing. ( a – d ) OptoHTS (left) and sOptoHTS (right) provide qualitatively and quantitatively similar results for measured effects on APD ( a , c ) and CTD ( b , d ) for both nifedipine ( a , b ) and dofetilide, a blocker of the rapid delayed-rectifier, I Kr ( c , d ). N =3–16 samples (at least 600 single-cell records or more) for each condition and each data point in the panels above. Data are presented as mean±s.e.m., and each well is considered an independent sample.

    Journal: Nature Communications

    Article Title: OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology

    doi: 10.1038/ncomms11542

    Figure Lengend Snippet: Experimental comparison of OptoHTS versus ‘spark'-OptoHTS for functional drug testing. ( a – d ) OptoHTS (left) and sOptoHTS (right) provide qualitatively and quantitatively similar results for measured effects on APD ( a , c ) and CTD ( b , d ) for both nifedipine ( a , b ) and dofetilide, a blocker of the rapid delayed-rectifier, I Kr ( c , d ). N =3–16 samples (at least 600 single-cell records or more) for each condition and each data point in the panels above. Data are presented as mean±s.e.m., and each well is considered an independent sample.

    Article Snippet: Drugs Nifedipine (MW 346.33 g mol−1 ; Sigma) concentrations of 50 μM, 10 μM, 5 μM, 1μM, 0.5 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM and 0.0001 μM were prepared in Tyrode's solution.

    Techniques: Functional Assay

    Demonstration of OptoHTS for HT dose–response drug testing. Nifedipine, an L-type Ca 2+ channel ( I CaL ) blocker, is applied at 12-concentration-graded dosing (0–50 μM) to ChR2-CMs in 96-well plates ( a ). Optical recordings of multiple voltage ( b – d ) or calcium events ( e – g ) are obtained during optical pacing at 1 Hz, screening the full plate in under 10 min (see also Supplementary Fig. 5 ). Example averaged over 10 s ( b , e ) and quantitative results for APD and CTD ( c , d , f , g ) are shown. Expected APD ( n =4–7 samples, at least 800 single-cell records per concentration) ( b – d ) and CTD ( n =4–6 samples, at least 800 single-cell records per concentration) ( e – g ) shortening, especially at the APD25/CTD25 and APD50/CTD50 levels, occurred due to nifedipine blocking the inward L-type calcium current. Maximum APD shortening is observed at around 1 μM, consistent with maximum block of I CaL reached at that concentration ( d , inset). Beyond 1 μM, indirect (voltage-mediated) or non-specific action on other ion channels partially counters the block of inward Ca 2+ current and can reduce or eliminate the APD shortening ( d ). Nifedipine appears to monotonically shorten CTD up to 10 μM ( f , g ). Data are presented as mean±s.e.m, and each well is considered an independent sample, represented by a spatially averaged trace.

    Journal: Nature Communications

    Article Title: OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology

    doi: 10.1038/ncomms11542

    Figure Lengend Snippet: Demonstration of OptoHTS for HT dose–response drug testing. Nifedipine, an L-type Ca 2+ channel ( I CaL ) blocker, is applied at 12-concentration-graded dosing (0–50 μM) to ChR2-CMs in 96-well plates ( a ). Optical recordings of multiple voltage ( b – d ) or calcium events ( e – g ) are obtained during optical pacing at 1 Hz, screening the full plate in under 10 min (see also Supplementary Fig. 5 ). Example averaged over 10 s ( b , e ) and quantitative results for APD and CTD ( c , d , f , g ) are shown. Expected APD ( n =4–7 samples, at least 800 single-cell records per concentration) ( b – d ) and CTD ( n =4–6 samples, at least 800 single-cell records per concentration) ( e – g ) shortening, especially at the APD25/CTD25 and APD50/CTD50 levels, occurred due to nifedipine blocking the inward L-type calcium current. Maximum APD shortening is observed at around 1 μM, consistent with maximum block of I CaL reached at that concentration ( d , inset). Beyond 1 μM, indirect (voltage-mediated) or non-specific action on other ion channels partially counters the block of inward Ca 2+ current and can reduce or eliminate the APD shortening ( d ). Nifedipine appears to monotonically shorten CTD up to 10 μM ( f , g ). Data are presented as mean±s.e.m, and each well is considered an independent sample, represented by a spatially averaged trace.

    Article Snippet: Drugs Nifedipine (MW 346.33 g mol−1 ; Sigma) concentrations of 50 μM, 10 μM, 5 μM, 1μM, 0.5 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM and 0.0001 μM were prepared in Tyrode's solution.

    Techniques: Concentration Assay, Blocking Assay

    Dynamic functional drug testing. A demonstration of the utility of OptoDyCE for spatio-temporal characterization. Dynamic pacing provides a means of studying pacing-induced V m and Ca 2+ restitution and instabilities ( a ), or drug-induced instabilities, that is, 2 μM dofetilide leading to voltage alternans at relatively low pacing frequency (2 Hz) ( b ). High-content dynamic information is obtained from a single data run ( c – h ). For example, restitution and temporal or spatial variability (quantified by MAD) are shown as function of both drug dose and pacing frequency for peak calcium in the presence of nifedipine ( c – e ) and for APD in the presence of dofetilide ( f , g ). Nifedipine action on peak calcium (per cent change) is dose-dependent but frequency-independent ( c , d ). Nifedipine appears to reduce temporal variability of peak calcium (assessed by MAD), and this reduction is augmented by higher frequency pacing ( e ). Dofetilide shows enhanced action on APD50 at higher frequency (opposite to reverse-use dependence) ( f , g ). Spatial variation as a function of drug dose can also be assessed by analysing multiple ROI within the same well ( h ) (see also Supplementary Figs 5–6 ). Dofetilide at 2 μM seems to increase spatial variability in APD, that is, increase dispersion of repolarization, compared with control during 1-Hz pacing ( P

    Journal: Nature Communications

    Article Title: OptoDyCE as an automated system for high-throughput all-optical dynamic cardiac electrophysiology

    doi: 10.1038/ncomms11542

    Figure Lengend Snippet: Dynamic functional drug testing. A demonstration of the utility of OptoDyCE for spatio-temporal characterization. Dynamic pacing provides a means of studying pacing-induced V m and Ca 2+ restitution and instabilities ( a ), or drug-induced instabilities, that is, 2 μM dofetilide leading to voltage alternans at relatively low pacing frequency (2 Hz) ( b ). High-content dynamic information is obtained from a single data run ( c – h ). For example, restitution and temporal or spatial variability (quantified by MAD) are shown as function of both drug dose and pacing frequency for peak calcium in the presence of nifedipine ( c – e ) and for APD in the presence of dofetilide ( f , g ). Nifedipine action on peak calcium (per cent change) is dose-dependent but frequency-independent ( c , d ). Nifedipine appears to reduce temporal variability of peak calcium (assessed by MAD), and this reduction is augmented by higher frequency pacing ( e ). Dofetilide shows enhanced action on APD50 at higher frequency (opposite to reverse-use dependence) ( f , g ). Spatial variation as a function of drug dose can also be assessed by analysing multiple ROI within the same well ( h ) (see also Supplementary Figs 5–6 ). Dofetilide at 2 μM seems to increase spatial variability in APD, that is, increase dispersion of repolarization, compared with control during 1-Hz pacing ( P

    Article Snippet: Drugs Nifedipine (MW 346.33 g mol−1 ; Sigma) concentrations of 50 μM, 10 μM, 5 μM, 1μM, 0.5 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM and 0.0001 μM were prepared in Tyrode's solution.

    Techniques: Functional Assay

    The effect of changing [Cl − ] o on EJPs and associated contractions evoked by sympathetic nerve stimulation The upper pair of traces ( A ) show the EJP and associated contraction initiated by three stimuli, recorded in the presence of nifedipine (1 μM). Three minutes after a 90 % replacement of [Cl − ] o with isethionate ions, the first component was dramatically reduced in amplitude but the second component and contraction were little changed ( B ). Some 9 min after reducing the [Cl − ] o , the first component was abolished, the second component persisted and the contraction was reduced in amplitude ( C ). The calibration bars refer to each set of traces.

    Journal: The Journal of Physiology

    Article Title: Sympathetic neuroeffector transmission in the rat anococcygeus muscle

    doi: 10.1111/j.1469-7793.1999.101aa.x

    Figure Lengend Snippet: The effect of changing [Cl − ] o on EJPs and associated contractions evoked by sympathetic nerve stimulation The upper pair of traces ( A ) show the EJP and associated contraction initiated by three stimuli, recorded in the presence of nifedipine (1 μM). Three minutes after a 90 % replacement of [Cl − ] o with isethionate ions, the first component was dramatically reduced in amplitude but the second component and contraction were little changed ( B ). Some 9 min after reducing the [Cl − ] o , the first component was abolished, the second component persisted and the contraction was reduced in amplitude ( C ). The calibration bars refer to each set of traces.

    Article Snippet: Drugs used in this study were guanethidine, prazosin hydrochloride, phentolamine mesylate, WB4101, ryanodine, nifedipine hydrochloride, Hepes buffer and caffeine (Sigma Chemicals).

    Techniques:

    Effect of caffeine and nifedipine on changes in [Ca 2+ ] i , membrane potential and contraction evoked by sympathetic nerve stimulation in the rat anococcygeus muscle A train of stimuli triggered a biphasic EJP ( Aa ), a transient increase in [Ca 2+ ] i ( Ab ) and a contraction ( Ac ). Caffeine (3 mM) reduced the amplitudes of the first component of the EJP ( Ba ), the increase in [Ca 2+ ] i ( Bb ) and almost abolished the contraction ( Bc ). The further addition of nifedipine (1 μM) abolished the increase in [Ca 2+ ] i ( Cb ), the associated contraction ( Cc ) and the remaining part of the first component but again left the second component of the EJP intact ( Ca ). The resting membrane potential in all traces was -74 mV. The time calibration bar refers to all traces. The voltage calibration bar refers to all membrane potential recordings, the fluorescence calibration bar applies to all ratiometric measurements and the force calibration bar applies to all recordings of contractions.

    Journal: The Journal of Physiology

    Article Title: Sympathetic neuroeffector transmission in the rat anococcygeus muscle

    doi: 10.1111/j.1469-7793.1999.101aa.x

    Figure Lengend Snippet: Effect of caffeine and nifedipine on changes in [Ca 2+ ] i , membrane potential and contraction evoked by sympathetic nerve stimulation in the rat anococcygeus muscle A train of stimuli triggered a biphasic EJP ( Aa ), a transient increase in [Ca 2+ ] i ( Ab ) and a contraction ( Ac ). Caffeine (3 mM) reduced the amplitudes of the first component of the EJP ( Ba ), the increase in [Ca 2+ ] i ( Bb ) and almost abolished the contraction ( Bc ). The further addition of nifedipine (1 μM) abolished the increase in [Ca 2+ ] i ( Cb ), the associated contraction ( Cc ) and the remaining part of the first component but again left the second component of the EJP intact ( Ca ). The resting membrane potential in all traces was -74 mV. The time calibration bar refers to all traces. The voltage calibration bar refers to all membrane potential recordings, the fluorescence calibration bar applies to all ratiometric measurements and the force calibration bar applies to all recordings of contractions.

    Article Snippet: Drugs used in this study were guanethidine, prazosin hydrochloride, phentolamine mesylate, WB4101, ryanodine, nifedipine hydrochloride, Hepes buffer and caffeine (Sigma Chemicals).

    Techniques: Fluorescence

    The effect of phentolamine, prazosin and yohimbine on EJPs and associated contractions evoked by sympathetic nerve stimulation Both the first and second components of the EJP, along with the contraction, were abolished by phentolamine (1 μm; A ). The resting membrane potential was -72 mV. In this preparation the stimuli initiated purinergic EJPs and these persisted in the presence of phentolamine. The middle pair of traces ( B ) show control responses and the effect of increasing concentrations of prazosin (0.3, 1 and 10 nM) on the membrane potential changes and contractions triggered by a pair of sympathetic stimuli. It can be seen that the lowest concentration applied (0.3 nM) abolished the contraction and reduced the amplitudes of both the first and second components of the EJP. Increasing the concentration of prazosin to 1 and 10 nM abolished the first but failed to abolish the second component. The resting membrane potential of this cell was -74 mV. The lower pair of traces ( C ) show control responses and the effect of increasing concentrations of yohimbine (10 and 100 nM) on the membrane potential changes and contractions triggered by three sympathetic stimuli. It can be seen that yohimbine (10 nM) reduced the amplitude of the first component but not that of the second component. Increasing the concentration of yohimbine to 100 nM reduced the amplitudes of both components but that of the first component more dramatically. The resting membrane potential of this cell was -75 mV. The time calibration applies to all recordings. Nifedipine (1 μM) was present throughout.

    Journal: The Journal of Physiology

    Article Title: Sympathetic neuroeffector transmission in the rat anococcygeus muscle

    doi: 10.1111/j.1469-7793.1999.101aa.x

    Figure Lengend Snippet: The effect of phentolamine, prazosin and yohimbine on EJPs and associated contractions evoked by sympathetic nerve stimulation Both the first and second components of the EJP, along with the contraction, were abolished by phentolamine (1 μm; A ). The resting membrane potential was -72 mV. In this preparation the stimuli initiated purinergic EJPs and these persisted in the presence of phentolamine. The middle pair of traces ( B ) show control responses and the effect of increasing concentrations of prazosin (0.3, 1 and 10 nM) on the membrane potential changes and contractions triggered by a pair of sympathetic stimuli. It can be seen that the lowest concentration applied (0.3 nM) abolished the contraction and reduced the amplitudes of both the first and second components of the EJP. Increasing the concentration of prazosin to 1 and 10 nM abolished the first but failed to abolish the second component. The resting membrane potential of this cell was -74 mV. The lower pair of traces ( C ) show control responses and the effect of increasing concentrations of yohimbine (10 and 100 nM) on the membrane potential changes and contractions triggered by three sympathetic stimuli. It can be seen that yohimbine (10 nM) reduced the amplitude of the first component but not that of the second component. Increasing the concentration of yohimbine to 100 nM reduced the amplitudes of both components but that of the first component more dramatically. The resting membrane potential of this cell was -75 mV. The time calibration applies to all recordings. Nifedipine (1 μM) was present throughout.

    Article Snippet: Drugs used in this study were guanethidine, prazosin hydrochloride, phentolamine mesylate, WB4101, ryanodine, nifedipine hydrochloride, Hepes buffer and caffeine (Sigma Chemicals).

    Techniques: Concentration Assay

    Effect of nifedipine on changes in [Ca 2+ ] i , membrane potential and contraction evoked by sympathetic nerve stimulation in the rat anococcygeus muscle Three impulses triggered a biphasic EJP ( Aa ), a transient and persistent increase in [Ca 2+ ] i ( Ab ) and a contraction ( Ac ). Nifedipine (1 μM) reduced the amplitudes of the initial component ( Ba), the change in [Ca 2+ ] i ( Bb ) and of the contraction ( Bc ) and abolished the persistent increase in [Ca 2+ ] i associated with the second component. The resting membrane potential in traces A and B was -71 mV. When [K + ] o was increased to 20 mM, the membrane potential depolarized by about 20 mV ( C ). Even though nifedipine was still present, the stimuli continued to initiate a biphasic EJP ( Ca), a transient increase in [Ca 2+ ] i ( Cb ) and a contraction ( Cc ). The time calibration bar applies to all traces. The voltage calibration bar refers to all membrane potential recordings, the fluorescence calibration bar applies to all ratiometric measurements and the force calibration bar applies to all recordings of contractions.

    Journal: The Journal of Physiology

    Article Title: Sympathetic neuroeffector transmission in the rat anococcygeus muscle

    doi: 10.1111/j.1469-7793.1999.101aa.x

    Figure Lengend Snippet: Effect of nifedipine on changes in [Ca 2+ ] i , membrane potential and contraction evoked by sympathetic nerve stimulation in the rat anococcygeus muscle Three impulses triggered a biphasic EJP ( Aa ), a transient and persistent increase in [Ca 2+ ] i ( Ab ) and a contraction ( Ac ). Nifedipine (1 μM) reduced the amplitudes of the initial component ( Ba), the change in [Ca 2+ ] i ( Bb ) and of the contraction ( Bc ) and abolished the persistent increase in [Ca 2+ ] i associated with the second component. The resting membrane potential in traces A and B was -71 mV. When [K + ] o was increased to 20 mM, the membrane potential depolarized by about 20 mV ( C ). Even though nifedipine was still present, the stimuli continued to initiate a biphasic EJP ( Ca), a transient increase in [Ca 2+ ] i ( Cb ) and a contraction ( Cc ). The time calibration bar applies to all traces. The voltage calibration bar refers to all membrane potential recordings, the fluorescence calibration bar applies to all ratiometric measurements and the force calibration bar applies to all recordings of contractions.

    Article Snippet: Drugs used in this study were guanethidine, prazosin hydrochloride, phentolamine mesylate, WB4101, ryanodine, nifedipine hydrochloride, Hepes buffer and caffeine (Sigma Chemicals).

    Techniques: Fluorescence

    Spontaneous neuronal calcium activity in GC-P and PGN-P in the RMS, RMS-OB and deep GCL. A. Schematic representation of an olfactory bulb sagittal slice. B. Ai14/GCaMP6s P0 animals received lateral or medial electroporation of a Cre mRNAs. C. Calcium activity was recorded on acute brain slices between 6 and 10 dpe. Red and green cells represent medial and lateral cells migrating in the RMS, RMS-OB or in the deep GCL. D.F.H. Raster plot of calcium activity of GC-P (left) and PGN-P (right) recorded in the RMS (D), RMS OB (F) and deep GCL (H). E.G.I. Percentage of active cells, frequency and mean amplitude for GC-P and PGN-P in the RMS (E), RMS-OB (G) and in the deep GCL (I). Cells were defined as active if they showed at least one detectable calcium transient. The calcium activity of GC-P greatly increased upon arrival in the RMS-OB and the deep GCL, in contrast with PGN-P activity. N = 6 slices per condition. Error bars are SEM. J. Left: Raster plot of calcium activity of GC-P in the deep GCL in control and calcium-free medium (No Calcium + 2mM EGTA). Right: The percentage of active cells is highly decreased in calcium free medium. K. Left: Raster plot of calcium activity of GC-P in the deep GCL in control condition and nifedipine (10 μM) supplemented medium. Right: The percentage of active after incubation with the L-Type Voltage Gated Calcium Channels antagonist Nifedipine at 10 μM is highly decreased.

    Journal: bioRxiv

    Article Title: Intrinsic neuronal activity during migration controls the recruitment of specific interneuron subtypes in the postnatal mouse olfactory bulb

    doi: 10.1101/2020.07.28.224568

    Figure Lengend Snippet: Spontaneous neuronal calcium activity in GC-P and PGN-P in the RMS, RMS-OB and deep GCL. A. Schematic representation of an olfactory bulb sagittal slice. B. Ai14/GCaMP6s P0 animals received lateral or medial electroporation of a Cre mRNAs. C. Calcium activity was recorded on acute brain slices between 6 and 10 dpe. Red and green cells represent medial and lateral cells migrating in the RMS, RMS-OB or in the deep GCL. D.F.H. Raster plot of calcium activity of GC-P (left) and PGN-P (right) recorded in the RMS (D), RMS OB (F) and deep GCL (H). E.G.I. Percentage of active cells, frequency and mean amplitude for GC-P and PGN-P in the RMS (E), RMS-OB (G) and in the deep GCL (I). Cells were defined as active if they showed at least one detectable calcium transient. The calcium activity of GC-P greatly increased upon arrival in the RMS-OB and the deep GCL, in contrast with PGN-P activity. N = 6 slices per condition. Error bars are SEM. J. Left: Raster plot of calcium activity of GC-P in the deep GCL in control and calcium-free medium (No Calcium + 2mM EGTA). Right: The percentage of active cells is highly decreased in calcium free medium. K. Left: Raster plot of calcium activity of GC-P in the deep GCL in control condition and nifedipine (10 μM) supplemented medium. Right: The percentage of active after incubation with the L-Type Voltage Gated Calcium Channels antagonist Nifedipine at 10 μM is highly decreased.

    Article Snippet: PharmacologyAntagonists were bath applied at the following concentrations: D-APV (Tocris Bioscience, 50μM), NBQX (Tocris Bioscience, 10μM), MPEP (Tocris Bioscience, 30μM), Bicuculline (Tocris Bioscience, 10μM), Strychnine (Tocris Bioscience, 20μM), Nifedipine (Tocris Bioscience, 10μM).

    Techniques: Activity Assay, Electroporation, Incubation

    Spontaneous neuronal calcium activity in GC-P in the deep GCL is not affected by classical antagonists. Incubation with a calcium free medium decrease the frequency of calcium transients. (A) The L-type voltage-dependent calcium channel blocker nifedipine decrease the frequency of calcium transients. (B) The NMDA receptors antagonist D-APV (C), AMPA/Kainate receptors antagonist NBQX (D), metabotropic glutamate receptor 5 antagonist MPEP (E), GABAA receptors antagonist Bicuculline (F) and glycine receptor antagonist strychnine (G) do no modify the percentage of active cells or the amplitude of calcium transient.

    Journal: bioRxiv

    Article Title: Intrinsic neuronal activity during migration controls the recruitment of specific interneuron subtypes in the postnatal mouse olfactory bulb

    doi: 10.1101/2020.07.28.224568

    Figure Lengend Snippet: Spontaneous neuronal calcium activity in GC-P in the deep GCL is not affected by classical antagonists. Incubation with a calcium free medium decrease the frequency of calcium transients. (A) The L-type voltage-dependent calcium channel blocker nifedipine decrease the frequency of calcium transients. (B) The NMDA receptors antagonist D-APV (C), AMPA/Kainate receptors antagonist NBQX (D), metabotropic glutamate receptor 5 antagonist MPEP (E), GABAA receptors antagonist Bicuculline (F) and glycine receptor antagonist strychnine (G) do no modify the percentage of active cells or the amplitude of calcium transient.

    Article Snippet: PharmacologyAntagonists were bath applied at the following concentrations: D-APV (Tocris Bioscience, 50μM), NBQX (Tocris Bioscience, 10μM), MPEP (Tocris Bioscience, 30μM), Bicuculline (Tocris Bioscience, 10μM), Strychnine (Tocris Bioscience, 20μM), Nifedipine (Tocris Bioscience, 10μM).

    Techniques: Activity Assay, Incubation

    Intracellular Ca 2+ signaling was reduced by L-803,087 in ganglion cells in retinal flat mount. A : applications of 52 mM K + solution for 30 s depolarized ganglion cells and produced transient increases in intracellular Ca 2+ concentration, measured with the dye fura-2. Traces ( left ) show that, in the absence of drug, the second of paired K + applications produced a smaller peak Ca 2+ signal. On the right , the paired K + pulses, with L-803,087 (100 nM) applied 30 s prior to the second K + application, show greater reduction of the Ca 2+ signal compared with control. Far right panel shows a fluorescent image (stimulated at 380 nm) of a fura-2-containing ganglion cell in the flat-mount retina. B : mean data show amplitudes of the second high K + response expressed as a percentage of first peak in control, L-803,087 (L-803, 100 nM), nifedipine (NIF, 10 μM), and nifedipine plus L-803,087. The mean Ca 2+ transient amplitudes of all 3 drug treatment groups (L-803, NIF, L-803 + NIF) were significantly smaller than those of the control group, but they did not differ among themselves ( n = 21–89 cells/group; * P

    Journal: Journal of Neurophysiology

    Article Title: Modulation of Voltage-Gated Ion Channels in Rat Retinal Ganglion Cells Mediated by Somatostatin Receptor Subtype 4

    doi: 10.1152/jn.00098.2010

    Figure Lengend Snippet: Intracellular Ca 2+ signaling was reduced by L-803,087 in ganglion cells in retinal flat mount. A : applications of 52 mM K + solution for 30 s depolarized ganglion cells and produced transient increases in intracellular Ca 2+ concentration, measured with the dye fura-2. Traces ( left ) show that, in the absence of drug, the second of paired K + applications produced a smaller peak Ca 2+ signal. On the right , the paired K + pulses, with L-803,087 (100 nM) applied 30 s prior to the second K + application, show greater reduction of the Ca 2+ signal compared with control. Far right panel shows a fluorescent image (stimulated at 380 nm) of a fura-2-containing ganglion cell in the flat-mount retina. B : mean data show amplitudes of the second high K + response expressed as a percentage of first peak in control, L-803,087 (L-803, 100 nM), nifedipine (NIF, 10 μM), and nifedipine plus L-803,087. The mean Ca 2+ transient amplitudes of all 3 drug treatment groups (L-803, NIF, L-803 + NIF) were significantly smaller than those of the control group, but they did not differ among themselves ( n = 21–89 cells/group; * P

    Article Snippet: L-803,087 (0.001%) and nifedipine (NIF, 0.1%) were prepared as DMSO stock solutions, frozen at −20°C, and thawed immediately before experiments (numbers in parentheses indicate percentage of DMSO in the final working solution).

    Techniques: Produced, Concentration Assay

    Effect of ion channel blockers on L55P TTR-induced calcium influx and analysis of sensitivity to icilin and capsaicin , (A) L55P was applied to DRG growth cones in the presence of VGCC inhibitors (nifedipine, ω-agatoxin IVA, ω-conotoxin GIVA), Na V inhibitors (tetrodotoxin, ambroxol and carbamazepine) and TRP inhibitors (SKF-96365, BCTC). The resulting maximal calcium influx (max ΔF/F 0 ) calculated over the imaging period (7 min) was calculated. (B) Effect of capsaicin (1 μM) and icilin (100 μM) on cytosolic calcium in DRG growth cones in culture. When DRG cultures were pre-treated with ambroxol (5 μM), icilin-induced calcium fluorescence was significantly decreased. All graphs show maximal ΔF/F 0 ± SEM for n = 12-24 growth cones. Significant differences from control values are depicted as: * p

    Journal: Molecular Neurodegeneration

    Article Title: TRPM8 and Nav1.8 sodium channels are required for transthyretin-induced calcium influx in growth cones of small-diameter TrkA-positive sensory neurons

    doi: 10.1186/1750-1326-6-19

    Figure Lengend Snippet: Effect of ion channel blockers on L55P TTR-induced calcium influx and analysis of sensitivity to icilin and capsaicin , (A) L55P was applied to DRG growth cones in the presence of VGCC inhibitors (nifedipine, ω-agatoxin IVA, ω-conotoxin GIVA), Na V inhibitors (tetrodotoxin, ambroxol and carbamazepine) and TRP inhibitors (SKF-96365, BCTC). The resulting maximal calcium influx (max ΔF/F 0 ) calculated over the imaging period (7 min) was calculated. (B) Effect of capsaicin (1 μM) and icilin (100 μM) on cytosolic calcium in DRG growth cones in culture. When DRG cultures were pre-treated with ambroxol (5 μM), icilin-induced calcium fluorescence was significantly decreased. All graphs show maximal ΔF/F 0 ± SEM for n = 12-24 growth cones. Significant differences from control values are depicted as: * p

    Article Snippet: Pharmacological compounds used in experiments were obtained from the following sources: nifedipine, ω-agatoxin IVA, ω-conotoxin GIVA (Alomone Labs, Israel), SKF-96365 (Tocris, Bristol, UK), [N-(4-tert-butylphenyl)-4-(3-chloropyridin-2-yl)piperazine-1-carboxamide] (BCTC), (ENZO, NY, USA), ambroxol and carbamazepine (Sigma-Aldrich, MO, USA).

    Techniques: Imaging, Fluorescence

    Voltage-dependent Ba 2+ currents in the epithelioid cells Epithelioid cells were voltage clamped at -70 mV in the presence of tetrodotoxin (0.2 μM) and BaCl 2 (5 mM) instead of CaCl 2 . A , representative recordings of current responses of an epithelioid cell to depolarizing potentials for 30 ms between -60 and +60 mV in 10 mV increments before (Control) and during exposure to CoCl 2 (2 mM), and after its removal (Washout). B , peak inward current-voltage relationships were obtained from the cell before (▪) and during (•) exposure to CoCl 2 , and after its removal (▴). C , effects of nifedipine and ω-conotoxin GVIA on peak Ba 2+ currents in an epithelioid cell. At -70 mV in the presence of tetrodotoxin (0.2 μM), TEA-Cl (5 mM) and BaCl 2 (5 mM), Ba 2+ currents were evoked by depolarizing steps for 15 ms to +10 mV with 20 s intervals. The peak Ba 2+ currents are expressed as percentages of the first Ba 2+ current recorded in the absence of blockers and plotted against time. Nifedipine (1 μM) and ω-conotoxin GVIA (1 μM) were applied during the periods indicated by horizontal lines starting with an arrow. Inset, superimposed current traces with letters corresponding to those in the peak current trace. The dotted line indicates the zero current level.

    Journal: The Journal of Physiology

    Article Title: Characteristics of 5-HT-containing chemoreceptor cells of the chicken aortic body

    doi: 10.1111/j.1469-7793.1999.049ad.x

    Figure Lengend Snippet: Voltage-dependent Ba 2+ currents in the epithelioid cells Epithelioid cells were voltage clamped at -70 mV in the presence of tetrodotoxin (0.2 μM) and BaCl 2 (5 mM) instead of CaCl 2 . A , representative recordings of current responses of an epithelioid cell to depolarizing potentials for 30 ms between -60 and +60 mV in 10 mV increments before (Control) and during exposure to CoCl 2 (2 mM), and after its removal (Washout). B , peak inward current-voltage relationships were obtained from the cell before (▪) and during (•) exposure to CoCl 2 , and after its removal (▴). C , effects of nifedipine and ω-conotoxin GVIA on peak Ba 2+ currents in an epithelioid cell. At -70 mV in the presence of tetrodotoxin (0.2 μM), TEA-Cl (5 mM) and BaCl 2 (5 mM), Ba 2+ currents were evoked by depolarizing steps for 15 ms to +10 mV with 20 s intervals. The peak Ba 2+ currents are expressed as percentages of the first Ba 2+ current recorded in the absence of blockers and plotted against time. Nifedipine (1 μM) and ω-conotoxin GVIA (1 μM) were applied during the periods indicated by horizontal lines starting with an arrow. Inset, superimposed current traces with letters corresponding to those in the peak current trace. The dotted line indicates the zero current level.

    Article Snippet: The solutions containing veratridine (Sigma), nifedipine (Wako Pure Chemicals, Japan) and Bay K 8644 (Sigma) were prepared by the addition of stock solutions of the drugs dissolved in DMSO.

    Techniques: Mass Spectrometry

    Effect of extracellular Ca 2+ removal, nifedipine, CPA, and ryanodine on PMA-induced airway SMC twitching. Line scans from airway wall regions obtained from phase-contrast images (similar to that presented in Fig. 1 ) showing airway SMC twitching induced by 10 µM PMA (added 30 min before the recordings started) and their sensitivity to: (A) Ca 2+ -free sHBSS, (B) 10 µM nifedipine, (C) 10 µM CPA, or (D) 25 µM ryanodine. PMA-induced SMC twitching was inhibited by removal of extracellular Ca 2+ , nifedipine, CPA, and ryanodine. Each line scan is representative of three experiments in lung slices from two mice.

    Journal: The Journal of General Physiology

    Article Title: Ca2+ oscillations, Ca2+ sensitization, and contraction activated by protein kinase C in small airway smooth muscle

    doi: 10.1085/jgp.201210876

    Figure Lengend Snippet: Effect of extracellular Ca 2+ removal, nifedipine, CPA, and ryanodine on PMA-induced airway SMC twitching. Line scans from airway wall regions obtained from phase-contrast images (similar to that presented in Fig. 1 ) showing airway SMC twitching induced by 10 µM PMA (added 30 min before the recordings started) and their sensitivity to: (A) Ca 2+ -free sHBSS, (B) 10 µM nifedipine, (C) 10 µM CPA, or (D) 25 µM ryanodine. PMA-induced SMC twitching was inhibited by removal of extracellular Ca 2+ , nifedipine, CPA, and ryanodine. Each line scan is representative of three experiments in lung slices from two mice.

    Article Snippet: GF-109203X and cyclopiazonic acid (CPA) were from Enzo Life Sciences, whereas Y-27632, nifedipine, and ryanodine were from Abcam.

    Techniques: Mouse Assay

    Effect of extracellular Ca 2+ removal, nifedipine, CPA, and ryanodine on PMA-induced Ca 2+ oscillations. Ca 2+ oscillations in single airway SMCs induced by 10 µM PMA (added 30 min before the recording started) and its inhibition by: (A) superfusion with Ca 2+ -free sHBSS, (B) 10 µM nifedipine, (C) 10 µM CPA, or (D) 25 µM ryanodine. Each trace is representative of four experiments in lung slices from three mice.

    Journal: The Journal of General Physiology

    Article Title: Ca2+ oscillations, Ca2+ sensitization, and contraction activated by protein kinase C in small airway smooth muscle

    doi: 10.1085/jgp.201210876

    Figure Lengend Snippet: Effect of extracellular Ca 2+ removal, nifedipine, CPA, and ryanodine on PMA-induced Ca 2+ oscillations. Ca 2+ oscillations in single airway SMCs induced by 10 µM PMA (added 30 min before the recording started) and its inhibition by: (A) superfusion with Ca 2+ -free sHBSS, (B) 10 µM nifedipine, (C) 10 µM CPA, or (D) 25 µM ryanodine. Each trace is representative of four experiments in lung slices from three mice.

    Article Snippet: GF-109203X and cyclopiazonic acid (CPA) were from Enzo Life Sciences, whereas Y-27632, nifedipine, and ryanodine were from Abcam.

    Techniques: Inhibition, Mouse Assay

    Displacement by nifedipine of [ 3 H]-R-PIA and [ 3 H]-XAC bound to membrane receptors. Open symbols [ 3 H]-R-PIA binding (K i app. 4.15 ± 1.37 × 10 −6 , B max app. 653 fmol/mg protein); closed symbols [ 3 H]-XAC (K i app. 1.86 ± 0.42 × 10 −6 M, B max app. 868 fmol/mg protein).

    Journal: Pharmacology & toxicology

    Article Title: Interaction of Dihydropyridine Calcium Channel Agonists and Antagonists with Adenosine Receptors

    doi:

    Figure Lengend Snippet: Displacement by nifedipine of [ 3 H]-R-PIA and [ 3 H]-XAC bound to membrane receptors. Open symbols [ 3 H]-R-PIA binding (K i app. 4.15 ± 1.37 × 10 −6 , B max app. 653 fmol/mg protein); closed symbols [ 3 H]-XAC (K i app. 1.86 ± 0.42 × 10 −6 M, B max app. 868 fmol/mg protein).

    Article Snippet: Methyl 2,6-dimethyl-5-nitro-4-(2-trifluoromethylphenyl) 1,4-dihydropyridine-3-carboxylate, Bay K 8644, and nifedipine were from Bayer AG.

    Techniques: Binding Assay

    High voltage activation calcium channels' L-type regulation and pathway signaling Ca 2+ /CaM-K II affect GH release in bovine somatotropes. (a) Bar graph illustrating the regulation of hormone release by GHRP-6 (100 nM) applied alone or in the presence of nifedipine (0.5 μ M) a blocker of HVACC L-type. (b) Average amount of GH released in the absences (control) and after treatment with the signaling pathway Ca 2+ /CaM-K II inhibitor KN-62 (10 μ M) alone or in combination with GHRP-6 (100 nM). Each value represents the mean ± SE of determinations performed in triplicate from three independent experiments. The asterisks denote significant differences ( P

    Journal: Journal of Signal Transduction

    Article Title: Upregulation of Voltage-Gated Calcium Channel Cav1.3 in Bovine Somatotropes Treated with Ghrelin

    doi: 10.1155/2013/527253

    Figure Lengend Snippet: High voltage activation calcium channels' L-type regulation and pathway signaling Ca 2+ /CaM-K II affect GH release in bovine somatotropes. (a) Bar graph illustrating the regulation of hormone release by GHRP-6 (100 nM) applied alone or in the presence of nifedipine (0.5 μ M) a blocker of HVACC L-type. (b) Average amount of GH released in the absences (control) and after treatment with the signaling pathway Ca 2+ /CaM-K II inhibitor KN-62 (10 μ M) alone or in combination with GHRP-6 (100 nM). Each value represents the mean ± SE of determinations performed in triplicate from three independent experiments. The asterisks denote significant differences ( P

    Article Snippet: Chemicals Ghrelin (ALX-157-022-M001), tetrodotoxin (BML-NA120-0001), KN-62 (BML-EI230), and nifedipine (ALX-550-091-G005) were purchased from Enzo-Alexis-Biomol (ENZO Life Sciences), GHRP-6 (HOR-298-1) was purchased from ProSpec protein specialist, and D-Lys3-GHRP-6 (G-4535-5MG) was purchased from Sigma (St. Louis, MO).

    Techniques: Activation Assay, Chick Chorioallantoic Membrane Assay

    Profiles of Relaxation by VDCC Blockers in Both WKY and SHR. Representative traces of nifedipine (A)-, verapamil (C)-, and Trp-His (E)-induced relaxation in PE (1 µM)-contracted aortic rings from both 8- and 40-week WKY and SHR. The relaxation activity was represented as EC 50 values for nifedipine (B), verapamil (D), and Trp-His (F). Results are expressed as the mean ± SEM values (n = 3–7). * P

    Journal: PLoS ONE

    Article Title: Attenuation of L-Type Ca2+ Channel Expression and Vasomotor Response in the Aorta with Age in Both Wistar-Kyoto and Spontaneously Hypertensive Rats

    doi: 10.1371/journal.pone.0088975

    Figure Lengend Snippet: Profiles of Relaxation by VDCC Blockers in Both WKY and SHR. Representative traces of nifedipine (A)-, verapamil (C)-, and Trp-His (E)-induced relaxation in PE (1 µM)-contracted aortic rings from both 8- and 40-week WKY and SHR. The relaxation activity was represented as EC 50 values for nifedipine (B), verapamil (D), and Trp-His (F). Results are expressed as the mean ± SEM values (n = 3–7). * P

    Article Snippet: Nifedipine was purchased from Nacalai Tesque (Kyoto, Japan).

    Techniques: Activity Assay

    Time-course analysis of drug absorption into PDMS, FEPM, and polystyrene (PS) culture plates (control). Drug concentrations after 0.5, 1, 2, 3, 6, and 24-h incubation of nifedipine, Bay K8644, and coumarin evaluated by high-performance liquid chromatography (HPLC, n = 3, * p

    Journal: Micromachines

    Article Title: Tetrafluoroethylene-Propylene Elastomer for Fabrication of Microfluidic Organs-on-Chips Resistant to Drug Absorption

    doi: 10.3390/mi10110793

    Figure Lengend Snippet: Time-course analysis of drug absorption into PDMS, FEPM, and polystyrene (PS) culture plates (control). Drug concentrations after 0.5, 1, 2, 3, 6, and 24-h incubation of nifedipine, Bay K8644, and coumarin evaluated by high-performance liquid chromatography (HPLC, n = 3, * p

    Article Snippet: Three types of drugs, nifedipine (N0528, Tokyo Chemical Industry, Tokyo, Japan), Bay K8644 (B112 Sigma-Aldrich, St. Louis, MO, USA), and coumarin (031-16562, Fujifilm Wako Chemicals, Osaka, Japan), were used to test drug absorption.

    Techniques: Incubation, High Performance Liquid Chromatography

    Curves for ABPM data plotted using the local regression LOESS function in R statistical software. Readings from 64 timepoints over 24 h for systolic and diastolic blood pressure (SBP and DBP) and heart rate (HR) provided 2,560 values for each parameter from 20 patients over two study periods. Solid‐red lines represent data fit from Mylan‐Nifedipine‐XL. Dashed‐green lines represent data fit from Nifedipine‐GITS. SBP curves for two patients are statistically different and continue to further separate during the terminal 8 h (nocturnal period) of the dosing interval, as delivery from Mylan‐Nifedipine‐XL is predicted to decline in a first‐order pattern. DBP curves follow a similar pattern, but are less affected by action of dihydropyridines and were not statistically different. HR remains clinically unaffected by constant, low‐rate delivery of nifedipine, varying by only 15 bpm over the course of the day.

    Journal: Clinical and Translational Science

    Article Title: Therapeutic Differences in 24‐h Ambulatory Blood Pressures in Patients Switched Between Bioequivalent Nifedipine Osmotic Systems With Differing Delivery Technologies

    doi: 10.1111/cts.12442

    Figure Lengend Snippet: Curves for ABPM data plotted using the local regression LOESS function in R statistical software. Readings from 64 timepoints over 24 h for systolic and diastolic blood pressure (SBP and DBP) and heart rate (HR) provided 2,560 values for each parameter from 20 patients over two study periods. Solid‐red lines represent data fit from Mylan‐Nifedipine‐XL. Dashed‐green lines represent data fit from Nifedipine‐GITS. SBP curves for two patients are statistically different and continue to further separate during the terminal 8 h (nocturnal period) of the dosing interval, as delivery from Mylan‐Nifedipine‐XL is predicted to decline in a first‐order pattern. DBP curves follow a similar pattern, but are less affected by action of dihydropyridines and were not statistically different. HR remains clinically unaffected by constant, low‐rate delivery of nifedipine, varying by only 15 bpm over the course of the day.

    Article Snippet: In conclusion, the observation of blood pressure differences in patients being switched between Nifedipine‐GITS and Mylan‐Nifedipine‐XL suggests that concerns about the adequacy of using current bioequivalence criteria to assess highly‐modified‐release formulations have a real foundation.

    Techniques: Software

    Plots of mixed‐effects fit of SBP data over time using GAMM function in R statistical software. Solid‐red lines represent data fit from Mylan‐Nifedipine‐XL. Dashed‐green lines represent data fit from Nifedipine‐GITS. There is a clear, statistically significant separation of 3 ± 1.1 mmHg ( P = 0.0173) between the curves of best population fit for each formulation, which was even larger during the last 8 h of the dosing interval.

    Journal: Clinical and Translational Science

    Article Title: Therapeutic Differences in 24‐h Ambulatory Blood Pressures in Patients Switched Between Bioequivalent Nifedipine Osmotic Systems With Differing Delivery Technologies

    doi: 10.1111/cts.12442

    Figure Lengend Snippet: Plots of mixed‐effects fit of SBP data over time using GAMM function in R statistical software. Solid‐red lines represent data fit from Mylan‐Nifedipine‐XL. Dashed‐green lines represent data fit from Nifedipine‐GITS. There is a clear, statistically significant separation of 3 ± 1.1 mmHg ( P = 0.0173) between the curves of best population fit for each formulation, which was even larger during the last 8 h of the dosing interval.

    Article Snippet: In conclusion, the observation of blood pressure differences in patients being switched between Nifedipine‐GITS and Mylan‐Nifedipine‐XL suggests that concerns about the adequacy of using current bioequivalence criteria to assess highly‐modified‐release formulations have a real foundation.

    Techniques: Software

    Flowchart of treatment assignment and timing of ambulatory blood pressure monitor (ABPM) assessments. Vertical arrows mark time of study initiation and ABPM measurements for either Mylan‐Nifedipine‐XL (Mylan) or Nifedipine‐GITS (N‐GITS).

    Journal: Clinical and Translational Science

    Article Title: Therapeutic Differences in 24‐h Ambulatory Blood Pressures in Patients Switched Between Bioequivalent Nifedipine Osmotic Systems With Differing Delivery Technologies

    doi: 10.1111/cts.12442

    Figure Lengend Snippet: Flowchart of treatment assignment and timing of ambulatory blood pressure monitor (ABPM) assessments. Vertical arrows mark time of study initiation and ABPM measurements for either Mylan‐Nifedipine‐XL (Mylan) or Nifedipine‐GITS (N‐GITS).

    Article Snippet: In conclusion, the observation of blood pressure differences in patients being switched between Nifedipine‐GITS and Mylan‐Nifedipine‐XL suggests that concerns about the adequacy of using current bioequivalence criteria to assess highly‐modified‐release formulations have a real foundation.

    Techniques:

    Neuritic degeneration induced by polybrene is dependent on Ca 2+ influx. a , b Ca 2+ influx induced by polybrene was inhibited by EGTA or nifedipine. Neurons were labeled by Fluo-4. a The time course of relative Fluo-4 fluorescence intensity was recorded. b The relative Fluo-4 fluorescence intensity at 3000 s. The fluorescence intensity of Fluo-4 at initiating time was normalized to 100 ( n ≥ 6, *** P

    Journal: Cell Death & Disease

    Article Title: Polybrene induces neural degeneration by bidirectional Ca2+ influx-dependent mitochondrial and ER–mitochondrial dynamics

    doi: 10.1038/s41419-018-1009-8

    Figure Lengend Snippet: Neuritic degeneration induced by polybrene is dependent on Ca 2+ influx. a , b Ca 2+ influx induced by polybrene was inhibited by EGTA or nifedipine. Neurons were labeled by Fluo-4. a The time course of relative Fluo-4 fluorescence intensity was recorded. b The relative Fluo-4 fluorescence intensity at 3000 s. The fluorescence intensity of Fluo-4 at initiating time was normalized to 100 ( n ≥ 6, *** P

    Article Snippet: Briefly, neurons were treated with polybrene for 24 h. For the polybrene treatment with 5 mM EGTA or 50 µM nifedipine (S1808, Selleck), drugs were added simultaneously with polybrene for 12 h. Neurons were observed under a phase-contrast microscope.

    Techniques: Labeling, Fluorescence

    mAChR agonist carbachol does not affect the depolarization-induced whole cell steady-state K Ca 1.1 currents under conditions of pharmacological inhibition of the major cellular sources of Ca 2+ for K Ca 1.1 channel activation in freshly-isolated human DSM cells A) Representative original recordings illustrate the depolarization-induced whole cell steady-state K Ca 1.1 currents in the absence (control) and in the presence of carbachol (1 μM). B) The current-voltage relationship curve summarizes the lack of carbachol effects on the whole cell steady-state K Ca 1.1 currents (n=9, N=6; P > 0.05). Steady-state K Ca 1.1 currents were recorded in the presence of ryanodine (30 μM), thapsigargin (100 nM), and nifedipine (1 μM). The bath solution contained 134 mM NaCl, 6 mM KCl, and 2 mM CaCl 2 , and the pipette 110 mM potassium aspartate, 30 mM KCl, 10 mM NaCl, and

    Journal: Pflugers Archiv : European journal of physiology

    Article Title: Functional link between muscarinic receptors and large-conductance Ca2+-activated K+ channels in freshly-isolated human detrusor smooth muscle cells

    doi: 10.1007/s00424-014-1537-8

    Figure Lengend Snippet: mAChR agonist carbachol does not affect the depolarization-induced whole cell steady-state K Ca 1.1 currents under conditions of pharmacological inhibition of the major cellular sources of Ca 2+ for K Ca 1.1 channel activation in freshly-isolated human DSM cells A) Representative original recordings illustrate the depolarization-induced whole cell steady-state K Ca 1.1 currents in the absence (control) and in the presence of carbachol (1 μM). B) The current-voltage relationship curve summarizes the lack of carbachol effects on the whole cell steady-state K Ca 1.1 currents (n=9, N=6; P > 0.05). Steady-state K Ca 1.1 currents were recorded in the presence of ryanodine (30 μM), thapsigargin (100 nM), and nifedipine (1 μM). The bath solution contained 134 mM NaCl, 6 mM KCl, and 2 mM CaCl 2 , and the pipette 110 mM potassium aspartate, 30 mM KCl, 10 mM NaCl, and

    Article Snippet: Stock solutions of ryanodine (Enzo Life Sciences, Inc.,NY), nifedipine (Thermo Fisher Scientific, NJ), thapsigargin (MP Biochemicals, LLC, CA), and paxilline (Sigma) were prepared in DMSO or ethanol.

    Techniques: Inhibition, Activation Assay, Isolation, Transferring

    Cumulative data showing the relative frequencies of LCR events during the peak of the responses to hyperosmotic sucrose, hyperosmotic CaCl 2 , or after transient exposure to a hypoosmotic solution in the absence or presence of nifedipine. ***, statistically significant differences (P

    Journal: The Journal of General Physiology

    Article Title: DHPR activation underlies SR Ca2+ release induced by osmotic stress in isolated rat skeletal muscle fibers

    doi: 10.1085/jgp.200910191

    Figure Lengend Snippet: Cumulative data showing the relative frequencies of LCR events during the peak of the responses to hyperosmotic sucrose, hyperosmotic CaCl 2 , or after transient exposure to a hypoosmotic solution in the absence or presence of nifedipine. ***, statistically significant differences (P

    Article Snippet: Stocks of 1 mM fura-2 AM (Biotium Inc.), 1 mM fluo-4 AM (Invitrogen), 1 mM tetracaine, 100 µM nifedipine (MP Biomedicals), 100 µM 9-AC (Alfa Aesar), 200 µM furosemide, or 1 mM bumetanide were dissolved in DMSO.

    Techniques:

    Representative confocal x-y images (1/second) obtained from fluo-4–loaded FDB fibers before, during, and after exposure to solutions of increased or decreased osmolarity. For each cell, the number of LCR events observed per frame throughout the experiment is indicated in the graph on the right. (A) The application of a hyperosmotic sucrose solution was often associated with a marked elevation of [Ca 2+ ], and LCR was difficult to identify (cell 1). However, where the rise in [Ca 2+ ] was less pronounced, individual LCR events were apparent during the application of sucrose (cell 2). Prior exposure to 100 µM nifedipine, to inhibit the DHPR, prevented LCR upon sucrose exposure (cell 3). (B) The introduction of a hyperosmotic CaCl 2 solution induced LCR (cell 1), which was markedly inhibited in the presence of nifedipine (cell 2). (C) The application of a hypoosmotic (254 mOsm) solution had no apparent effect on [Ca 2+ ] i . However, returning to the isoosmotic solution precipitated LCR events (cell 1). In the presence of nifedipine, transient exposure to a hypoosmotic solution failed to induce LCR (cell 2). Fibers subject to hypoosmotic Tyrode's exposure were initially exposed to isoosmotic Tyrode for 30 s, and then for 2 min to a hypoosmotic before returning to isoosmotic Tyrode's solution for 4 min. Horizontal bar, 20 µm.

    Journal: The Journal of General Physiology

    Article Title: DHPR activation underlies SR Ca2+ release induced by osmotic stress in isolated rat skeletal muscle fibers

    doi: 10.1085/jgp.200910191

    Figure Lengend Snippet: Representative confocal x-y images (1/second) obtained from fluo-4–loaded FDB fibers before, during, and after exposure to solutions of increased or decreased osmolarity. For each cell, the number of LCR events observed per frame throughout the experiment is indicated in the graph on the right. (A) The application of a hyperosmotic sucrose solution was often associated with a marked elevation of [Ca 2+ ], and LCR was difficult to identify (cell 1). However, where the rise in [Ca 2+ ] was less pronounced, individual LCR events were apparent during the application of sucrose (cell 2). Prior exposure to 100 µM nifedipine, to inhibit the DHPR, prevented LCR upon sucrose exposure (cell 3). (B) The introduction of a hyperosmotic CaCl 2 solution induced LCR (cell 1), which was markedly inhibited in the presence of nifedipine (cell 2). (C) The application of a hypoosmotic (254 mOsm) solution had no apparent effect on [Ca 2+ ] i . However, returning to the isoosmotic solution precipitated LCR events (cell 1). In the presence of nifedipine, transient exposure to a hypoosmotic solution failed to induce LCR (cell 2). Fibers subject to hypoosmotic Tyrode's exposure were initially exposed to isoosmotic Tyrode for 30 s, and then for 2 min to a hypoosmotic before returning to isoosmotic Tyrode's solution for 4 min. Horizontal bar, 20 µm.

    Article Snippet: Stocks of 1 mM fura-2 AM (Biotium Inc.), 1 mM fluo-4 AM (Invitrogen), 1 mM tetracaine, 100 µM nifedipine (MP Biomedicals), 100 µM 9-AC (Alfa Aesar), 200 µM furosemide, or 1 mM bumetanide were dissolved in DMSO.

    Techniques:

    (A) Simultaneous records of E m (top) and the fura-2 fluorescence ratio (bottom) from fibers equilibrated with an isoosmotic solution before transient exposure to solutions made hyperosmotic (404 mOsm) by the addition of sucrose, mannitol, or CaCl 2 , or hypoosmotic (254 mOsm) by decreasing [NaCl] (left-right). (B) The effects of hyperosmotic sucrose or CaCl 2 solutions on E m (top) and the fura-2 fluorescence ratio (bottom) in cells exposed to 100 µM nifedipine to inhibit DHPR activation. (C) Substitution of 100 mM Na + for K + resulted in a sustained depolarization. Thereafter, the introduction of hyperosmotic sucrose solution had no apparent effect on [Ca 2+ ] i , whereas the application of 30 mM caffeine induced a robust [Ca 2+ ] i transient. (D) Accumulated data showing both the relative change in fluorescence ratio (filled bars) and E m (open bars) obtained using the protocols shown in A and B. Bars represent the mean (± SEM), and the number of preparations is indicated in parentheses. **, mean values significantly different to the peak values obtained after the introduction of hyperosmotic sucrose (P

    Journal: The Journal of General Physiology

    Article Title: DHPR activation underlies SR Ca2+ release induced by osmotic stress in isolated rat skeletal muscle fibers

    doi: 10.1085/jgp.200910191

    Figure Lengend Snippet: (A) Simultaneous records of E m (top) and the fura-2 fluorescence ratio (bottom) from fibers equilibrated with an isoosmotic solution before transient exposure to solutions made hyperosmotic (404 mOsm) by the addition of sucrose, mannitol, or CaCl 2 , or hypoosmotic (254 mOsm) by decreasing [NaCl] (left-right). (B) The effects of hyperosmotic sucrose or CaCl 2 solutions on E m (top) and the fura-2 fluorescence ratio (bottom) in cells exposed to 100 µM nifedipine to inhibit DHPR activation. (C) Substitution of 100 mM Na + for K + resulted in a sustained depolarization. Thereafter, the introduction of hyperosmotic sucrose solution had no apparent effect on [Ca 2+ ] i , whereas the application of 30 mM caffeine induced a robust [Ca 2+ ] i transient. (D) Accumulated data showing both the relative change in fluorescence ratio (filled bars) and E m (open bars) obtained using the protocols shown in A and B. Bars represent the mean (± SEM), and the number of preparations is indicated in parentheses. **, mean values significantly different to the peak values obtained after the introduction of hyperosmotic sucrose (P

    Article Snippet: Stocks of 1 mM fura-2 AM (Biotium Inc.), 1 mM fluo-4 AM (Invitrogen), 1 mM tetracaine, 100 µM nifedipine (MP Biomedicals), 100 µM 9-AC (Alfa Aesar), 200 µM furosemide, or 1 mM bumetanide were dissolved in DMSO.

    Techniques: Fluorescence, Activation Assay

    Predicted and observed (A) plasma concentration profile and (B) change in systolic blood pressure after a single dose of nifedipine 60 mg GITS in North European hypertensive subjects ( Meredith and Elliott, 2004 ). Predicted and observed (C) plasma concentration profile and (D) change in systolic blood pressure after the initial dose and daily dosing of nifedipine 30 mg GITS for 15 days in North European hypertensive subjects ( Brown and Toal, 2008 ).

    Journal: Frontiers in Pharmacology

    Article Title: Applications of linking PBPK and PD models to predict the impact of genotypic variability, formulation differences, differences in target binding capacity and target site drug concentrations on drug responses and variability

    doi: 10.3389/fphar.2014.00258

    Figure Lengend Snippet: Predicted and observed (A) plasma concentration profile and (B) change in systolic blood pressure after a single dose of nifedipine 60 mg GITS in North European hypertensive subjects ( Meredith and Elliott, 2004 ). Predicted and observed (C) plasma concentration profile and (D) change in systolic blood pressure after the initial dose and daily dosing of nifedipine 30 mg GITS for 15 days in North European hypertensive subjects ( Brown and Toal, 2008 ).

    Article Snippet: The PK and PD profiles for nifedipine in the treatment of hypertension were simulated using the Simcyp nifedipine compound file ( Table ), a minimal PBPK distribution model and elimination by enzyme kinetics.

    Techniques: Concentration Assay

    Representative traces of single N-channel activity recorded from cell-attached patches of control NG108-15 and cav1(+) cells. ( A ) Traces from a control NG108-15 cell obtained with pulses from −40 to +20 mV of 180 ms each. The averaged current at the bottom (−0.051 pA) was calculated over 257 sweeps including nulls. ( B ) Traces from a cav1(+) cell. The averaged current at the bottom is 69% smaller (−0.016 pA) than control (423 sweeps including nulls). The patch pipette contained 100 mM Ba 2+ and the L-type Ca 2+ channel blocker nifedipine (10 μ M).

    Journal: Biophysical Journal

    Article Title: Caveolin-1 Expression and Membrane Cholesterol Content Modulate N-Type Calcium Channel Activity in NG108-15 Cells

    doi: 10.1529/biophysj.105.065623

    Figure Lengend Snippet: Representative traces of single N-channel activity recorded from cell-attached patches of control NG108-15 and cav1(+) cells. ( A ) Traces from a control NG108-15 cell obtained with pulses from −40 to +20 mV of 180 ms each. The averaged current at the bottom (−0.051 pA) was calculated over 257 sweeps including nulls. ( B ) Traces from a cav1(+) cell. The averaged current at the bottom is 69% smaller (−0.016 pA) than control (423 sweeps including nulls). The patch pipette contained 100 mM Ba 2+ and the L-type Ca 2+ channel blocker nifedipine (10 μ M).

    Article Snippet: All experiments in cell-attached patches were carried out with the recording pipette containing 100 mM Ba2+ and 10 μ M nifedipine.

    Techniques: Activity Assay, Mass Spectrometry, Transferring