RT-PCR for detecting ion channels expressed in human cardiac fibroblasts.

<p><i>A.</i> Images of RT-PCR products corresponding to significant gene expression of KCa1.1 (BK_Ca), Kv1.5 (IK_DR), Kv4.3 (I_to), and Kir2.1 (I_Kir) and Clcn3 (I_Cl.vol), and Na_V1.2, Na_V1.3, Na_V1.5, Na_V1.6 and Na_V1.7 in human cardiac fibroblasts. A weak expression of Kv4.2, Kir2.3, Clcn2 and Na_V1.1 was also found in human cardiac fibroblasts. <i>B.</i> No significant bands were observed in the PCR experiment when RT product was replaced by total RNA.</p

I_to in human cardiac fibroblasts.

<p><i>A.</i> I_to traces recorded in a representative cell with the voltage protocol showed in the <i>inset</i> in the absence and presence of 5 mM 4-AP. <i>B.</i> Normalized mean values of voltage-dependent availability (I/I_max) and activation conductance (g/g_max) of I_to were fitted to the Boltzmann function: y = 1/{1+exp[(V_m−V_0.5)/S]}, where V_m is membrane potential, V_0.5 is the estimated midpoint, and S is the slope factor. <i>C.</i> Normalized I_to (I₂/I₁) plotted vs. P₁−P₂ interval. The recovery curve was fitted to a mono-exponential function. The I_to was measured from the current peak to the ‘quasi’-steady-state level.</p

Effect of Ba²⁺ on membrane current in human cardiac fibroblasts.

<p><i>A.</i> Voltage-dependent currents were reversibly inhibited by 0.5 mM BaCl₂ in a representative cell. Currents were recorded with the protocol as shown in the <i>inset</i> (0.2 Hz). <i>B.</i> Voltage-dependent current recorded in another cell with voltage protocol shown in the <i>inset</i> of A was increased by elevating K⁺_o from 5 to 20 mM. Ba²⁺ (0.5 mM) remarkably suppressed the current. <i>C.</i> Left panel: <i>I-V</i> relationships of membrane currents recorded in a representative cell with a 2-s ramp protocol (−120 to 0 mV from a holding potential of −40 mV) in 5 mM K⁺_o, 20 mM K⁺_o, and after application of 0.5 mM Ba²⁺. Right panel: Ba²⁺-sensitive <i>I-V</i> relationships of the membrane current, typical of I_Kir.</p

I_Na.TTX and I_Na.TTXR in human cardiac fibroblasts.

<p><i>A.</i> An inward current with a persistent component (arrow) recorded in a representative cell under K⁺-free conditions using the voltage steps as shown in the <i>inset</i>. Nifedipine (10 µM) had no effect on the current, while the current disappeared when Na⁺_o was replaced with equimolar choline, and recovered as restoration of Na⁺_o. <i>B.</i> Similar inward current with persistent component (arrow) recorded in another cell was highly sensitive to inhibition by low concentrations of TTX. <i>C.</i> An inward current with fast inactivation recorded using the same voltage protocol as shown in the <i>inset</i> of A. The current was not affected by 10 nM TTX, but reversibly inhibited by 10 µM nifedipine. <i>D.</i> Similar current recorded in another cell disappeared with Na⁺_o removal, and recovered as restoration of Na⁺_o. The current was suppressed by a high concentration of TTX (10 µM). <i>E.</i> Concentration-dependent response of two types of inward currents to TTX. The data were fitted to the Hill equation: E = E_max/[1+(IC₅₀/C)^b], where E is the percentage inhibition of current at concentration C, E_max is the maximum inhibition, IC₅₀ is the concentration for a half inhibitory effect, and b is the Hill coefficient. The IC₅₀ of TTX for inhibiting TTX-sensitive I_Na was 7.8 nM (n = 5−9 for each concentration), the Hill coefficient was 0.94. The IC₅₀ of TTX for inhibiting TTX-resistant I_Na was 1.8 µM (n = 6−9 cell for each concentration), the Hill coefficient was 0.58. <i>F.</i> Concentration-dependent relationships of I_Na.TTX and I_Na.TTXR to nifedipine. The IC₅₀ of nifedipine for inhibiting I_Na.TTXR was 56.2 µM (n = 4−7 cells for each concentration) with a Hill coefficient of 0.59.</p

I_Cl in human cardiac fibroblasts.

<p>A. Voltage-dependent current was inhibited by the Cl⁻ channel blocker DIDS (150 µM). Current was elicited by the voltage steps as shown in the <i>inset</i> (0.2 Hz). <i>B. </i><i> I-V</i> relation curve of DIDS-sensitive current obtained by subtracting currents before and after DIDS application in A. <i>C.</i> Voltage-dependent current recorded in a representative cells during control, after 20 min 0.7T exposure and application of 100 µM NPPB. <i>D.</i><i> I-V</i> relationships for control current (1.0T), 0.7T and 0.7T with 100 µM NPPB. The 0.7T-induced current was significantly inhibited by NPPB at all test potentials (n<i> = </i>5, P<0.01). The arrows in the figure indicate the zero current level.</p

BK_Ca and IK_DR in human cardiac fibroblasts.

<p><i>A.</i> Voltage-dependent current was reversibly suppressed by the BK_Ca blocker paxilline (1 µM). Currents were elicited by the voltage protocol as shown in the <i>inset</i>. <i>B.</i> Current-voltage (<i>I-V</i>) relationships of membrane current were recorded by a 2-s ramp protocol (−80 to +80 mV from a holding potential −40 mV) in a representative cell in the absence and presence of 1 µM paxilline. <i>C.</i> Membrane currents recorded in a typical experiment with the same voltage protocol as in A were partially inhibited by 1 µM paxilline. The remaining current was suppressed by co-application of paxilline and 5 mM 4-AP.</p

Families of membrane currents in human cardiac fibroblasts.

<p><i>A.</i> Noisy current was activated at positive potential. Currents were elicited with the protocol shown in the <i>inset</i> (0.2 Hz). <i>B.</i> A transient outward current was activated in a human cardiac fibroblast by the same protocol as in A. <i>C.</i> A current with inward rectification activated by hyperpolarized potentials (<i>inset</i>) was co-present with the noisy current. <i>D.</i> Voltage-dependent current with outward rectification was recorded with the same protocol as in C. <i>E.</i> An inward current with fast inactivation activated by depolarization voltage steps (<i>inset</i>) was co-present with the noisy current. <i>F.</i> An inward current with slow inactivation (arrow) activated by the same protocol as in E was co-present with the noisy current.</p

Kinetics of I_Na.TTX and I_Na.TTXR.

<p><i>A.</i> Mean values of <i>I-V</i> relationships of I_Na.TTX and I_Na.TTXR. <i>B.</i> Left panel: inactivation time course of representative I_Na traces (at 0 mV) was fitted to a monoexponential function with time constant (τ) shown, 4.3 ms for I_Na.TTX and 1.82 ms for I_Na.TTXR. Right panel: mean values of voltage dependence of inactivation of I_Na.TTX (n = 8) and I_Na.TTXR (n = 10). P<0.05 or P<0.01 at −20 to +60 mV. <i>C.</i> Voltage-dependent availability (I/I_max) of I_Na was determined with the protocol as shown in the left <i>inset</i> (with 1-s conditioning pulses from voltages between −120 and −10 mV then a 50-ms test pulse to 0 mV). Curves of I/I_max and activation conductance (g/g_max) were fitted to a Boltzmann equation. <i>E.</i> Recovery curves of I_Na.TTX and I_Na.TTXR from inactivation were fitted to a monoexponential function.</p

Additional file 1: of PR interval prolongation in coronary patients or risk equivalent: excess risk of ischemic stroke and vascular pathophysiological insights

Table S1. Univariable and Multivariable Predictors for Carotid IMT. Table S2. Univariable and Multivariable Predictors for Cardiovascular Death. Table S3. Univariable and Multivariable Predictors for New-Onset Ischemic Stroke. Table S4. Univariable and Multivariable Predictors for New-Onset Myocardial Infarction. Table S5. Univariable and Multivariable Predictors for Combined Cardiovascular Endpoints of New-Onset Myocardial Infarction, Ischemic Stroke, Congestive Heart Failure and Cardiovascular Death. Table S6. Estimates of Sensitivity (Se), Specificity (Sp), Positive Predictive Value (PPV) and Negative Predictive Value (NPV) of PR Interval in the Prediction for Cardiovascular Events. (DOC 444 kb

Det är knappast ofarligt att demonstrera i Sverige

<p>(A) Kaplan-Meier curves for thromboembolic event-free survival. Log-rank: 19.714. <i>P</i><0.001. (B) Annual incidence of thromboembolic events. CHA₂DS₂-VASc = 1: 0.54% per year (95% CI 0.45–0.67); CHA₂DS₂-VASc = 2–3: 1.54% per year (95% CI 1.41–1.70); CHA₂DS₂-VASc = 4–5: 2.98% per year (95% CI 2.81–3.18); CHA₂DS₂-VASc ≥6: 5.04% per year (95% CI 4.59–5.60).</p