Electrophysiologic effects of the IK1 inhibitor PA-6 are modulated by extracellular potassium in isolated guinea pig hearts

The pentamidine analog PA‐6 was developed as a specific inward rectifier potassium current (I(K) (1)) antagonist, because established inhibitors either lack specificity or have side effects that prohibit their use in vivo. We previously demonstrated that BaCl(2), an established I(K) (1) inhibitor, could prolong action potential duration (APD) and increase cardiac conduction velocity (CV). However, few studies have addressed whether targeted I(K) (1) inhibition similarly affects ventricular electrophysiology. The aim of this study was to determine the effects of PA‐6 on cardiac repolarization and conduction in Langendorff‐perfused guinea pig hearts. PA‐6 (200 nm) or vehicle was perfused into ex‐vivo guinea pig hearts for 60 min. Hearts were optically mapped with di‐4‐ANEPPS to quantify CV and APD at 90% repolarization (APD (90)). Ventricular APD (90) was significantly prolonged in hearts treated with PA‐6 (115 ± 2% of baseline; P < 0.05), but not vehicle (105 ± 2% of baseline). PA‐6 slightly, but significantly, increased transverse CV by 7%. PA‐6 significantly prolonged APD (90) during hypokalemia (2 mmol/L [K+](o)), although to a lesser degree than observed at 4.56 mmol/L [K+](o). In contrast, the effect of PA‐6 on CV was more pronounced during hypokalemia, where transverse CV with PA‐6 (24 ± 2 cm/sec) was significantly faster than with vehicle (13 ± 3 cm/sec, P < 0.05). These results show that under normokalemic conditions, PA‐6 significantly prolonged APD (90), whereas its effect on CV was modest. During hypokalemia, PA‐6 prolonged APD (90) to a lesser degree, but profoundly increased CV. Thus, in intact guinea pig hearts, the electrophysiologic effects of the I(K) (1) inhibitor, PA‐6, are [K+](o)‐dependent

InAs/MoRe hybrid semiconductor/superconductor nanowire devices

Implementing superconductors capable of proximity-inducing a large energy-gap in semiconductors in the presence of strong magnetic fields is a major goal towards applications of semiconductor/superconductor hybrid materials in future quantum information technologies. Here, we study the performance of devices consisting of InAs nanowires in electrical contact to molybdenum-rhenium (MoRe) superconducting alloys. The MoRe thin films exhibit transition temperatures ~10 K and critical fields exceeding 6 T. Normal/superconductor devices enabled tunnel spectroscopy of the corresponding induced superconductivity, which was maintained up to ~10 K, and MoRe based Josephson devices exhibit supercurrents and multiple Andreev reflections. We determine an induced superconducting gap lower than expected from the transition temperature, and observe gap softening at finite magnetic field. These may be common features for hybrids based on large gap, type-II superconductors. The results encourage further development of MoRe-based hybrids

Engineering Hybrid Epitaxial InAsSb/Al Nanowire Materials for Stronger Topological Protection

The combination of strong spin-orbit coupling, large $g$-factors, and the coupling to a superconductor can be used to create a topologically protected state in a semiconductor nanowire. Here we report on growth and characterization of hybrid epitaxial InAsSb/Al nanowires, with varying composition and crystal structure. We find the strongest spin-orbit interaction at intermediate compositions in zincblende InAs$_{1-x}$Sb$_{x}$ nanowires, exceeding that of both InAs and InSb materials, confirming recent theoretical studies \cite{winkler2016topological}. We show that the epitaxial InAsSb/Al interfaces allows for a hard induced superconducting gap and 2$e$ transport in Coulomb charging experiments, similar to experiments on InAs/Al and InSb/Al materials, and find measurements consistent with topological phase transitions at low magnetic fields due to large effective $g$-factors. Finally we present a method to grow pure wurtzite InAsSb nanowires which are predicted to exhibit even stronger spin-orbit coupling than the zincblende structure.Comment: 10 pages and 5 figure

The KCNQ1 potassium channel is down-regulated by ubiquitylating enzymes of the Nedd4/Nedd4-like family

Objective: The voltage-gated KCNQ1 potassium channel regulates key physiological functions in a number of tissues. In the heart, KCNQ1 α-subunits assemble with KCNE1 β-subunits forming a channel complex constituting the delayed rectifier current IKs. In epithelia, KCNQ1 channels participate in controlling body electrolyte homeostasis. Several regulatory mechanisms of the KCNQ1 channel complexes have been reported, including protein kinase A (PKA)-phosphorylation and β-subunit interactions. However, the mechanisms controlling the membrane density of KCNQ1 channels have attracted less attention. Methods and results: Here we demonstrate that KCNQ1 proteins expressed in HEK293 cells are down-regulated by Nedd4/Nedd4-like ubiquitin-protein ligases. KCNQ1 and KCNQ1/KCNE1 currents were reduced upon co-expression of Nedd4-2, the isoform among the nine members of the Nedd4/Nedd4-like family displaying the highest expression level in human heart. In vivo expression of a catalytically inactive form of Nedd4-2, able to antagonize endogenous Nedd4-2 in guinea-pig cardiomyocytes, increased IKs significantly, but did not modify IK1. Concomitant with the reduction in current induced by Nedd4-2, an increased ubiquitylation as well as a decreased total level of KCNQ1 proteins were observed in HEK293 cells. Pull-down and co-immunoprecipitation experiments showed that Nedd4-2 interacts with the C-terminal part of KCNQ1. The Nedd4/Nedd4-like-mediated regulation of the KCNQ1 channel complexes is strictly dependent on a PY motif located in the distal part of the C-terminal domain. When this motif was mutated, the current and ubiquitylation levels were unaffected by Nedd4-2, and Nedd4-2 proteins were neither pulled-down nor co-immunoprecipitated. Conclusions: These results suggest that KCNQ1 internalization and stability is physiologically regulated by its Nedd4/Nedd4-like-dependent ubiquitylation. This mechanism may thereby be important in regulating the surface density of the KCNQ1 channels in cardiomyocytes and other cell type

Antiarrhythmic and electrophysiologic effects of flecainide on acutely induced atrial fibrillation in healthy horses

BACKGROUND: Only few pharmacologic compounds have been validated for treatment of atrial fibrillation (AF) in horses. Studies investigating the utility and safety of flecainide to treat AF in horses have produced conflicting results, and the antiarrhythmic mechanisms of flecainide are not fully understood. OBJECTIVES: To study the potential of flecainide to terminate acutely induced AF of short duration (≥15 minutes), to examine flecainide‐induced changes in AF duration and AF vulnerability, and to investigate the in vivo effects of flecainide on right atrial effective refractory period, AF cycle length, and ventricular depolarization and repolarization. ANIMALS: Nine Standardbred horses. Eight received flecainide, 3 were used as time‐matched controls, 2 of which also received flecainide. METHODS: Prospective study. The antiarrhythmic and electrophysiologic effects of flecainide were based on 5 parameters: ability to terminate acute pacing‐induced AF (≥15 minutes), and drug‐induced changes in atrial effective refractory period, AF duration, AF vulnerability, and ventricular depolarization and repolarization times. Parameters were assessed at baseline and after flecainide by programmed electrical stimulation methods. RESULTS: Flecainide terminated all acutely induced AF episodes (n = 7); (AF duration, 21 ± 5 minutes) and significantly decreased the AF duration, but neither altered atrial effective refractory period nor AF vulnerability significantly. Ventricular repolarization time was prolonged between 8 and 20 minutes after initiation of flecainide infusion, but no ventricular arrhythmias were detected. CONCLUSIONS AND CLINICAL IMPORTANCE: Flecainide had clear antiarrhythmic properties in terminating acute pacing‐induced AF, but showed no protective properties against immediate reinduction of AF. Flecainide caused temporary prolongation in the ventricular repolarization, which may be a proarrhythmic effect

Milestones toward Majorana-based quantum computing

We introduce a scheme for preparation, manipulation, and read out of Majorana zero modes in semiconducting wires with mesoscopic superconducting islands. Our approach synthesizes recent advances in materials growth with tools commonly used in quantum-dot experiments, including gate control of tunnel barriers and Coulomb effects, charge sensing, and charge pumping. We outline a sequence of milestones interpolating between zero-mode detection and quantum computing that includes (1) detection of fusion rules for non-Abelian anyons using either proximal charge sensors or pumped current, (2) validation of a prototype topological qubit, and (3) demonstration of non-Abelian statistics by braiding in a branched geometry. The first two milestones require only a single wire with two islands, and additionally enable sensitive measurements of the system’s excitation gap, quasiparticle poisoning rates, residual Majorana zero-mode splittings, and topological-qubit coherence times. These pre-braiding experiments can be adapted to other manipulation and read out schemes as well

Cellulose filtration of blood from malaria patients for improving <i>ex vivo</i> growth of <i>Plasmodium falciparum</i> parasites

BACKGROUND: Establishing in vitro Plasmodium falciparum culture lines from patient parasite isolates can offer deeper understanding of geographic variations of drug sensitivity and mechanisms of malaria pathogenesis and immunity. Cellulose column filtration of blood is an inexpensive, rapid and effective method for the removal of host factors, such as leucocytes and platelets, significantly improving the purification of parasite DNA in a blood sample. METHODS: In this study, the effect of cellulose column filtration of venous blood on the initial in vitro growth of P. falciparum parasite isolates from Tanzanian children admitted to hospital was tested. The parasites were allowed to expand in culture without subcultivation until 5 days after admission or the appearance of dead parasites and parasitaemia was determined daily. To investigate whether the filtration had an effect on clonality, P. falciparum merozoite surface protein 2 genotyping was performed using nested PCR on extracted genomic DNA, and the var gene transcript levels were investigated, using quantitative PCR on extracted RNA, at admission and 4 days of culture. RESULTS: The cellulose-filtered parasites grew to higher parasitaemia faster than non-filtered parasites seemingly due to a higher development ratio of ring stage parasites progressing into the late stages. Cellulose filtration had no apparent effect on clonality or var gene expression; however, evident differences were observed after only 4 days of culture in both the number of clones and transcript levels of var genes compared to the time of admission. CONCLUSIONS: Cellulose column filtration of parasitized blood is a cheap, applicable method for improving cultivation of P. falciparum field isolates for ex vivo based assays; however, when assessing phenotype and genotype of cultured parasites, in general, assumed to represent the in vivo infection, caution is advised. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12936-017-1714-2) contains supplementary material, which is available to authorized users