Family pedigree, clinical evaluation, and molecular genetics.

<p>(<b>A</b>) The index patient (III-1) is indicated by an arrow. Individuals indicated with black squares/circles carry the mutation and a clinical phenotype (III-1, III-2, II-2). Individuals indicated with grey circles (II-3 to II-5) were clinically diagnosed with DCM, but not genotyped. Abbreviation: DCM (dilated cardiomyopathy). (<b>B</b>) 12-lead ECG of the index patient showing third degree AV-block with a ventricular escape rhythm and a small QRS-complex with a heart rate of 43 bpm (artefact in lead V1). (<b>C</b>) Non-sustained ventricular tachycardia (220 bpm) occurred at a heart rate of 130 bpm and a work load of 192 W during an exercise stress test. (<b>D</b>) Different DHPLC eluting profiles at 59.8°C of the PCR products of exon 6 in the index patient compared to the control. Abbreviation: DHPLC (denaturing high performance liquid chromatography). (<b>E</b>) A heterozygous change of arginine CGC (R) to histidine CAC (H) resulted in the missense mutation R219H. (<b>F</b>) Sequence alignments of the S4 of domain 1 from Na⁺ and K⁺ (<i>Shaker</i> B) channels in different species.</p

Proton current-voltage relationship of the Na_v1.5/R219H channel recorded in an NMDG Na⁺-free solution.

<p>(<b>A</b>) Representative proton current traces from oocytes expressing the Na_v1.5/R219H channel recorded at pH_o 8.40, 7.40, 6.80, and 6.00, as indicated, in response to 200 ms voltage steps ranging from −140 mV to +40 mV in 5-mV increments from a holding potential of −80 mV (the protocol is given in the centre inset), without on-line leak subtraction. The dashed line represents the zero current. For clarity, only current every 10 mV are shown. (<b>B</b>) Current-voltage relationship where the currents in (<b>A</b>) were plotted as a function of the test potential (5 mV increments), after offline linear leak subtraction. Reversal potential determined in a Na⁺-free NMDG solution at pH_o 8.40 using voltage steps as described in (<b>A</b>). The pH_i was measured using a pH-sensitive electrode. Similar results were obtained with four separate batches of oocytes. The inset shows the pH_oand pH_i values and between parentheses is the predicted values calculated using the Nernst equation. The bleu trace shows the voltage-dependent of activation (Q–V), the grey zone illustrates the transitional zone corresponding to the probability of the voltage sensor being stabilized in the outward position. (<b>C</b>) Correlation between the peak Na⁺ current measured in Ringer's solution and the proton current measured at −140 mV and pH_o 4.00 (n = 31) on the same oocytes. The data were obtained from one batch of oocytes over three days. The straight line represents the linear regression of the data set and R² is the correlation coefficient and shows the goodness of fit. Similar results were obtained with three separate batches of oocytes. (<b>D</b>) Proton currents measured in response to a change in pH_o at −140 mV in an NMDG Na⁺-free solution. The currents were normalized to the currents obtained at pH_o = 4.00 for each cell. The mean data (n = 5) was fitted to the Henderson-Hasselbach equation, 1/[1+exp(2.3(pH_o−pK_a))]. Error bars are smaller than the symbols.</p

Na_v1.5/R219H induces an inward proton current and intracellular acidification.

<p><i>Xenopus</i> oocytes expressing Na_v1.5/WT or Na_v1.5/R219H channel were impaled with three electrodes, one filled with an H⁺ resin to measure pH_i, and two to clamp the oocyte at −80 mV in a Na⁺-free NMDG solution containing 1 µM TTX, as indicated. Typical proton current recordings (red traces) in response to different pH_o value and the pH_i measurement rate (bleu traces) from an oocyte expressing the Na_v1.5/R219H (<b>A</b>) or Na_v1.5/WT channel (<b>B</b>). Intracellular pH_i values before changing solutions in experiments similar to (<b>A</b>) and (<b>B</b>) were plotted against pH_o (***, p<0.001 compared to WT, n = 10–19)(<b>C</b>). Similar recordings were obtained with four batches of oocytes. (<b>D</b>) Changes in pH_i after incubating oocytes expressing the Na_v1.5/WT (triangles) or Na_v1.5/R219H (squares) channel, or water-injected oocytes (circles) in OR3 medium at different pH_o values (***, p<0.001, **; p<0.01; *, p<0.05; compared to WT, n = 7–13). pH_i measurements were carried out in Ringer's solution at pH_o of 7.40.</p

Biophysical characterization of the Na_v1.5/R219H DCM mutation proton current recordings.

<p>Representative current traces recorded using the cut-open oocyte technique from Na_v1.5/WT (<b>A</b>) and Na_v1.5/R219H (<b>B</b>) channels. Currents were elicited by depolarizing pulses from −100 mV to +60 mV, with 10 mV increments for each step. (<b>C</b>) The voltage dependence of steady-state activation and inactivation of WT (activation, n = 7; inactivation, n = 8) and R219H (activation, n = 8; inactivation, n = 8). Activation curves were derived from <i>I</i>–<i>V</i> curves and fitted to a standard Boltzmann equation: <i>G</i> (<i>V</i>)/<i>G </i>_max = 1/(1+exp ((<i>V</i>−<i>V</i>_1/2)/<i>k_v</i>)), with midpoints (V_1/2) is slow factors (<i>k_v</i>) listed in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038331#pone.0038331.s001" target="_blank">Table S1</a></b>. The voltage-dependence of inactivation was induced by applying conditioning pre-pulses to membrane potentials ranging from a holding potential of −150 to −20 mV for 500 ms with 10 mV increments and was then measured using a 20-ms test pulse to −30 mV for each step (see protocol in inset). The recorded inactivation data were fitted to a standard Boltzmann equation: <i>I</i> (<i>V</i>)/<i>I</i>_max = 1/(1+exp ((<i>V</i>−<i>V</i>_1/2)/<i>k_v</i>)), with midpoints (<i>V</i>_1/2) is slow factors (<i>k_v</i>) listed in <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038331#pone.0038331.s001" target="_blank">Table S1</a></b>. (<b>D</b>) Time courses of recovery from inactivation of Na_v1.5/WT and Na_v1.5/R219H channels. A 40 ms conditioning pre-pulse was used to monitor recovery using a 20-ms test pulse after a variable recovery interval ranging from 5 to 500 ms (see protocol in inset). A single-exponential function was used to determine the time constants of recovery.</p

Procedural data.

<p>Procedural data.</p

Baseline characteristics.

<p>Baseline characteristics.</p

(A) Anterior-posterior view of the right atrial map including the path of the ablation catheter into the left atrium. Note that the color of the proximal ring electrodes of the ablation catheter have switched from the normal grey to black indicating their position within the sheath, thereby ensuring left atrial position of the transseptal sheath. (B) The ablation catheter is then exchanged for the circular mapping catheter to obtain a map of the left atrium.

<p>The ablation catheter is then inserted into the second transseptal sheath and left atrial access is gained based on the known location and orientation of the PFO. The white line and the three white tags delineate the mitral annulus. Note that the right atrial map is switched to a different color (green) to clearly appreciate the edges of the two maps.</p

(A) Antero-posterior view of the fast anatomical map of the inferior vena cava. While advancing the catheter up to the right atrium, catheter tip orientation, force vector orientation, and contact force are monitored (outlined box). (B) Left anterior oblique view of the map of the right atrium with the 3D MRI reconstruction of the left atrium positioned based on the course of the coronary sinus and the septum.

<p>The yellow tag denotes the His position.</p

Table_1_Complement and endothelial cell activation in COVID-19 patients compared to controls with suspected SARS-CoV-2 infection: A prospective cohort study.docx

BackgroundThromboinflammation may influence disease outcome in COVID-19. We aimed to evaluate complement and endothelial cell activation in patients with confirmed COVID-19 compared to controls with clinically suspected but excluded SARS-CoV-2 infection.MethodsIn a prospective, observational, single-center study, patients presenting with clinically suspected COVID-19 were recruited in the emergency department. Blood samples on presentation were obtained for analysis of C5a, sC5b-9, E-selectin, Galectin-3, ICAM-1 and VCAM-1.Results153 cases and 166 controls (suffering mainly from non-SARS-CoV-2 respiratory viral infections, non-infectious inflammatory conditions and bacterial pneumonia) were included. Hospital admission occurred in 62% and 45% of cases and controls, respectively. C5a and VCAM-1 concentrations were significantly elevated and E-selectin concentrations decreased in COVID-19 out- and inpatients compared to the respective controls. However, relative differences in outpatients vs. inpatients in most biomarkers were comparable between cases and controls. Elevated concentrations of C5a, Galectin-3, ICAM-1 and VCAM-1 on presentation were associated with the composite outcome of ICU- admission or 30-day mortality in COVID-19 and controls, yet more pronounced in COVID-19. C5a and sC5b-9 concentrations were significantly higher in COVID-19 males vs. females, which was not observed in the control group.ConclusionsOur data indicate an activation of the complement cascade and endothelium in COVID-19 beyond a nonspecific inflammatory trigger as observed in controls (i.e., “over”-activation).</p

Datasheet1_Effects of SARS-COV-2 infection on outcomes in patients hospitalized for acute cardiac conditions. A prospective, multicenter cohort study (Swiss Cardiovascular SARS-CoV-2 Consortium).pdf

BackgroundAlthough the severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) causing coronavirus disease 2019 (COVID-19) primarily affects the respiratory system, the disease entity has been associated with cardiovascular complications. This study sought to assess the effect of concomitant SARS-COV-2 infection on clinical outcomes of patients hospitalized primarily for acute cardiac conditions on cardiology wards in Switzerland.MethodsIn this prospective, observational study conducted in 5 Swiss cardiology centers during the COVID-19 pandemic, patients hospitalized due to acute cardiac conditions underwent a reverse-transcriptase polymerase chain reaction test at the time of admission and were categorized as SARS-COV-2 positive (cases) or negative (controls). Patients hospitalized on cardiology wards underwent treatment for the principal acute cardiac condition according to local practice. Clinical outcomes were recorded in-hospital, at 30 days, and after 1 year and compared between cases and controls. To adjust for imbalanced baseline characteristics, a subgroup of patients derived by propensity matching was analyzed.ResultsBetween March 2020 and February 2022, 538 patients were enrolled including 122 cases and 416 controls. Mean age was 68.0 ± 14.7 years, and 75% were men. Compared with controls, SARS-COV-2-positive patients more commonly presented with acute heart failure (35% vs. 17%) or major arrhythmia (31% vs. 9%), but less commonly with acute coronary syndrome (26% vs. 53%) or severe aortic stenosis (4% vs. 18%). Mortality was significantly higher in cases vs. controls in-hospital (16% vs. 1%), at 30 days (19.0% vs. 2.2%), and at 1 year (28.7% vs. 7.6%: p ConclusionsIn this observational study of patients hospitalized for acute cardiac conditions, SARS-COV-2 infection at index hospitalization was associated with markedly higher all-cause and non-cardiovascular mortality throughout one-year follow-up. These findings highlight the need for effective, multifaceted management of both cardiac and non-cardiac morbidities and prolonged surveillance in patients with acute cardiac conditions complicated by SARS-COV-2 infection.</p