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Methods for one-photon fluorescent imaging of calcium dynamics in vivo are popular due to their ability to simultaneously capture the dynamics of hundreds of neurons across large fields of view, at a low equipment complexity and cost. In contrast to two-photon methods, however, one-photon methods suffer from higher levels of crosstalk between cell bodies and the surrounding neuropil, resulting in decreased signal-to-noise and artifactual correlations of neural activity. Here, we address this problem by engineering cell body-targeted variants of the fluorescent calcium indicator GCaMP6f. We screened fusions of GCaMP6f to both natural as well as engineered peptides, and identified fusions that localized GCaMP6f to within approximately 50 microns of the cell body of neurons in live mice and larval zebrafish. One-photon imaging of soma-targeted GCaMP6f in dense neural circuits reported fewer artifactual spikes from neuropil, increased signal-to-noise ratio, and decreased artifactual correlation across neurons. Thus, soma-targeting of fluorescent calcium indicators increases neuronal signal fidelity and may facilitate even greater usage of simple, powerful, one-photon methods of population imaging of neural calcium dynamics.

Methods for highly multiplexed RNA imaging are limited in spatial resolution, and thus in their ability to localize transcripts to nanoscale and subcellular compartments. We adapt expansion microscopy, which physically expands biological specimens, for long-read untargeted and targeted in situ RNA sequencing. We applied untargeted expansion sequencing (ExSeq) to mouse brain, yielding readout of thousands of genes, including splice variants and novel transcripts. Targeted ExSeq yielded nanoscale-resolution maps of RNAs throughout dendrites and spines in neurons of the mouse hippocampus, revealing patterns across multiple cell types; layer-specific cell types across mouse visual cortex; and the organization and position-dependent states of tumor and immune cells in a human metastatic breast cancer biopsy. Thus ExSeq enables highly multiplexed mapping of RNAs, from nanoscale to system scale.One Sentence SummaryIn situ sequencing of physically expanded specimens enables multiplexed mapping of RNAs at nanoscale, subcellular resolution.

In a revision of Berberis Sect. Wallichianae in Taiwan and Luzon, the Philippines, two of the authors recognized 11 species from Taiwan, including three new species. This article recognizes a further new species, Berberis morii, and places it in the same clade with other species from Taiwan. Rather than reproduce all the observations that were made there, we simply note that Taiwan is an important center of endemism of B. Sect. Wallichianae. In comparing the floral structure and overall morphology of B. morii with similar taxa from Taiwan, China and India, a series morphological differences, both qualitatively and quantitatively, were noted. Phylogenetic analysis was also used to evaluate the endemicity of B. morii.

As has long been known Berberis Sect. Wallichianae in Taiwan evidence a high species diversity all of which are endemic to the island. Though cytological data has been used to understand adaptive speciation for certain Berberis species elsewhere, little is still known about the chromosome numbers of Taiwanese Berberis. In this study, the chromosomes of eleven Taiwanese Berberis Sect. Wallichianae were counted for the first time and were all determined to be diploids (2n = 28). This result not only reconfirms previous studies elsewhere which have concluded that polyploidization may not be a dominant evolutionary process in Berberis but also suggests that geographic isolation or other extrinsic factors may play a more important role on the rapid speciation of Berberis in Taiwan.

We explored the temporal and spatial diversification of the plant genus Sedum L. (Crassulaceae) in Taiwan based on molecular analysis of nrITS and cpDNA sequences from East Asian Sedum members. Our phylogenetic and ancestral area reconstruction analysis showed that Taiwanese Sedum comprised two lineages that independently migrated from Japan and Eastern China. Furthermore, the genetic distances among species in these two clades were smaller than those of other East Asian Sedum clades, and the Taiwanese members of each clade occupy extremely varied habitats with similar niches in high-mountain regions. These data indicate that species diversification occurred in parallel in the two Taiwanese Sedum lineages, and that these parallel radiations could have occurred within the small continental island of Taiwan. Moreover, the estimated time of divergence for Taiwanese Sedum indicates that the two radiations might have been correlated to the formation of mountains in Taiwan during the early Pleistocene. We suggest that these parallel radiations may be attributable to the geographical dynamics of Taiwan and specific biological features of Sedum that allow them to adapt to new ecological niches.

Introduction A novel amyloid β (Aβ) synthetic peptide vaccine (UB-311) has been evaluated in a first-in-human trial with patients of mild-to-moderate Alzheimer's disease. We describe translational research covering vaccine design, preclinical characterization, and phase-I clinical trial with supportive outcome that advances UB-311 into an ongoing phase-II trial. Methods UB-311 is constructed with two synthetic Aβ1–14–targeting peptides (B-cell epitope), each linked to different helper T-cell peptide epitopes (UBITh®) and formulated in a Th2-biased delivery system. The hAPP751 transgenic mouse model was used to perform the proof-of-concept study. Baboons and macaques were used for preclinical safety, tolerability, and immunogenicity evaluation. Patients with mild-to-moderate Alzheimer's disease (AD) were immunized by intramuscular route with 3 doses of UB-311 at weeks 0, 4, and 12, and monitored until week 48. Safety and immunogenicity were assessed per protocol, and preliminary efficacy was analyzed by Alzheimer's Disease Assessment Scale–Cognitive Subscale (ADAS-Cog), Mini–Mental State Examination (MMSE), and Alzheimer's Disease Cooperative Study–Clinician's Global Impression of Change (ADCS-CGIC). Results UB-311 covers a diverse genetic background and facilitates strong immune response with high responder rate. UB-311 reduced the levels of Aβ1–42 oligomers, protofibrils, and plaque load in hAPP751 transgenic mice. Safe and well-tolerated UB-311 generated considerable site-specific (Aβ1–10) antibodies across all animal species examined. In AD patients, UB-311 induced a 100% responder rate; injection site swelling and agitation were the most common adverse events (4/19 each). A slower rate of increase in ADAS-Cog from baseline to week 48 was observed in the subgroup of mild AD patients (MMSE ≥ 20) compared with the moderate AD subgroup, suggesting that UB-311 may have a potential of cognition improvement in patients with early stage of Alzheimer's dementia. Discussion The UBITh® platform can generate a high-precision molecular vaccine with high responder rate, strong on-target immunogenicity, and a potential of cognition improvement, which support UB-311 for active immunotherapy in early-to-mild AD patients currently enrolled in a phase-II trial (NCT02551809).

Visual Abstract, Highlights • In systolic heart failure, decreased renal perfusion due to impaired cardiac pumping activates the renal nerves, which send a signal to the brain to call for help. • The brain thus activates the neurohormonal system to increase organ perfusion, which may predispose the heart to ventricular arrhythmia. • Chemical renal denervation with phenol cuts the signal sent to the brain and thus decreases the susceptibility to ventricular arrhythmia in rats with systolic heart failure., Summary Several studies have shown the beneficial effect of renal denervation (RDN) in the treatment of ventricular arrhythmia, especially in the setting of heart failure (HF). However, the underlying mechanism of antiarrhythmic effect of RDN is unknown. Arrhythmogenic cardiac alternans, particularly spatially discordant repolarization alternans, characterized by simultaneous prolongation and shortening of action potential duration (APD) in different myocardial regions, is central to the genesis of ventricular fibrillation in HF. Whether RDN decreases the susceptibility to arrhythmogenic cardiac alternans in HF has never been addressed before. The authors used a rat model of post-myocardial infarction HF and dual voltage-calcium optical mapping to investigate whether RDN could attenuate arrhythmogenic cardiac alternans that predisposes to ventricular arrhythmias, as well as the hemodynamic effect of RDN in HF. The HF rats had increased body weights, dilated hearts, and lower blood pressure. The HF rats also had longer ventricular APDs and a delay in the decay of the calcium transient, typical electrophysiological features of human HF. Susceptibility to calcium transient alternans, APD alternans, and spatially discordant APD alternans was increased in the HF hearts. RDN significantly attenuated a delay in the decay of the calcium transient, calcium transient and APD alternans, and importantly, the discordant APD alternans, and thereby decreased the incidence of induced ventricular arrhythmia in HF. RDN did not further decrease blood pressure in HF rats. In conclusion, RDN improves calcium cycling and prevents spatially discordant APD alternans and ventricular arrhythmia in HF. RDN does not aggravate hemodynamics in HF.

Mutant allele specific imbalance (MASI) was initially coined to describe copy number alterations associated with the mutant allele of an oncogene. The copy number gain (CNG) specific to the mutant allele can be readily observed in electropherograms. With the development of genome-wide analyses at base-pair resolution with copy number counts, we can now further differentiate MASI into those with CNG, with copy neutral alteration (also termed acquired uniparental disomy; UPD), or with loss of heterozygosity (LOH) due to the loss of the wild-type (WT) allele. Here we summarize the occurrence of MASI with CNG, aUPD, or MASI with LOH in some major oncogenes (such as EGFR, KRAS, PIK3CA, and BRAF). We also discuss how these various classifications of MASI have been demonstrated to impact tumorigenesis, progression, metastasis, prognosis, and potentially therapeutic responses in cancer, notably in lung, colorectal, and pancreatic cancers.

Expansion microscopy (ExM) physically magnifies biological specimens to enable nanoscale-resolution imaging on conventional microscopes. Current ExM methods permeate biological specimens with free radical-polymerized polyacrylate hydrogels, whose network structure limits the microscopy resolution enabled by ExM. Here we report that ExM is possible using hydrogels with more homogeneous network structure, assembled via non-radical terminal linking of monomers of tetrahedral shape. As with earlier forms of ExM, such “tetra-gel”-embedded specimens can be iteratively expanded for greater physical magnification. Iterative tetra-gel expansion of herpes simplex virus type 1 (HSV-1) virions by ~10x in linear dimension results in a viral envelope deviation from sphericity of 9.2 nm, rather than the 14.3 nm enabled by free radical-polymerized hydrogels used in earlier versions of ExM. Thus, tetra-gel polymer chemistry may support new forms of ExM imaging that introduce fewer spatial errors than earlier versions, and raise the question of whether single biomolecule precision may be achievable.

Expansion microscopy (ExM) enables imaging of preserved specimens with nanoscale precision on diffraction-limited instead of specialized super-resolution microscopes. ExM works by physically separating fluorescent probes after anchoring them to a swellable gel. The first ExM method did not result in the retention of native proteins in the gel and relied on custom-made reagents that are not widely available. Here we describe protein retention ExM (proExM), a variant of ExM in which proteins are anchored to the swellable gel, allowing the use of conventional fluorescently labeled antibodies and streptavidin, and fluorescent proteins. We validated and demonstrated the utility of proExM for multicolor super-resolution (∼70 nm) imaging of cells and mammalian tissues on conventional microscopes.

Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia. Previous genome-wide association studies had identified single-nucleotide polymorphisms in several genomic regions to be associated with AF. In human genome, copy number variations (CNVs) are known to contribute to disease susceptibility. Using a genome-wide multistage approach to identify AF susceptibility CNVs, we here show a common 4,470-bp diallelic CNV in the first intron of potassium interacting channel 1 gene (KCNIP1) is strongly associated with AF in Taiwanese populations (odds ratio=2.27 for insertion allele; P=6.23 × 10−24). KCNIP1 insertion is associated with higher KCNIP1 mRNA expression. KCNIP1-encoded protein potassium interacting channel 1 (KCHIP1) is physically associated with potassium Kv channels and modulates atrial transient outward current in cardiac myocytes. Overexpression of KCNIP1 results in inducible AF in zebrafish. In conclusions, a common CNV in KCNIP1 gene is a genetic predictor of AF risk possibly pointing to a functional pathway., Tsai et al. here utilize a multi-stage genome-wide association study in Taiwanese population to show a copy number variation in the intron of potassium interacting channel 1 gene (KCNIP1) to be strongly associated with atrial fibrillation. The study also examines the functionality of KCNIP1 in heart electrophysiological function using cultured myocytes and zebrafish.

Objectives Whether the spironolactone treatment remains effective for the prevention of atrial fibrillation (AF) in dialysis patients is unclear. Methods We used a database from the Registry for Catastrophic Illness from the National Health Research Institute. All dialysis patients aged 18 or older without history of AF before ESRD were incorporated. A total of 113,191 dialysis patients were enrolled in the study. The median follow-up time was 4.17years. We collected information on prescribed drug dosage, number of days of treatment and the total number of pills dispensed from the outpatient pharmacy prescription database. All individuals in the study cohort with the first occurrence of AF were included as cases. Results In spironolactone group, the incidence of developing new AF was significantly lower than that in the control group both before (0.8% vs. 3.3%, P=0.019) and after PS matching (1.2% vs. 3.0%, P=0.019). Before PS matching, Cox's proportional hazard regression analyses showed that spironolactone was associated with 60% reduction of new AF (HR=0.372 [0.200–0.692], P=0.002) and the protective effect is dose-responsive in accumulated dose, treatment duration and mean daily dose. After PS matching, the overall AF prevention effect remained significant (HR=0.400 [0.179–0.895], P=0.026) while the dose–response relationship became borderline significant. Subgroup analyses showed that the protective effect was more evident in some specific subgroup patients. Conclusion Our study showed that spironolactone therapy was associated with lower risk of developing AF in a dose-responsive manner in patients with dialysis. Further randomized study is needed to confirm this observation.

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AIMS This was a randomized controlled study performed to compare 8 mm-tip catheter cryoablation (CRYO) with radiofrequency ablation (RFA) in treating atrioventricular nodal re-entrant tachycardia (AVNRT). METHODS AND RESULTS A total of 158 patients (103 women, mean age 48.9 ± 14.1) with symptomatic AVNRT (140 typical; 18 atypical) were randomized to undergo CRYO with an 8 mm-tip catheter (n = 80) or RFA (n = 78). The primary endpoint was a composite of acute procedural failure, inadvertent permanent atrioventricular block (AVB) and recurrence at 12 months. No significant difference was observed between CRYO and RFA groups in primary endpoint (7.5 vs. 11.5%; P = 0.764), 12-month recurrence rate (3.8 vs. 1.3%; P = 0.358), inadvertent permanent AVB (0 vs. 1.3%; P = 0.307), and acute procedural failure (3.7 vs. 9%; P = 0.128). In patients with acute procedure failure, success was achieved in 5 of 7 patients (71.4%) in RFA group and 2 of 3 patients (66.7%) in CRYO group on cross-over. There was no significant difference in procedural duration between CRYO and RFA groups (72.4 ± 41.6 vs. 63.7 ± 29.8 min; P = 0.13), but fluoroscopic duration in CRYO group was significantly shorter (3.4 ± 6.3 vs. 6.7 ± 7.4 min; P = 0.005). Patient pain score (2.7 ± 2.7 vs. 4.6 ± 2.7; P

Background/Purpose Brugada syndrome (BrS) is a hereditable sudden cardiac death (SCD). Mutations in the SCN5A gene (the most common BrS-causing gene) are responsible for 20–25% of this disease in Caucasian populations. However, the prevalence of SCN5A mutations in patients with BrS in the Chinese Han population in Taiwan remains unknown. Therefore, in this study, we investigated the prevalence of the SCN5A mutation in the largest BrS cohort in Taiwan. Methods We consecutively enrolled 47 unrelated patients with BrS from medical centers and hospitals in Taiwan between 2000 and 2010. Mutations within all the 27 translated exons, and exon–intron boundaries of the SCN5A -encoded cardiac sodium channel were screened in all patients with BrS using direct sequencing. A total of 500 unrelated healthy volunteers with a normal electrocardiogram were genotyped as a control group. Results SCN5A genetic variants were identified in 14 of the 47 patients with BrS and four of the 14 patients with BrS had missense mutations (1651 G>A, 1776 C>G, 3578 G>A). The prevalence rate of SCN5A mutations was approximately 8% (4/47), which was significantly lower than that reported in Caucasian populations (20–25%; p = 0.0007). The average age of these 14 BrS patients with SCN5A variants at diagnosis (12 men and 2 women) was 40 ± 13 years. Four patients experienced SCD, and six presented with seizure or syncope. Only three patients (3/14, 21.4%) had a family history of SCD. Conclusion The prevalence of SCN5A mutations in the Chinese Han population in Taiwan may be lower than that reported in the Caucasian populations. In addition, most patients with BrS did not have a family history of SCD.

Background— The transmural distribution of apamin-sensitive small conductance Ca 2+ -activated K + (SK) current ( I KAS ) in failing human ventricles remains unclear. Methods and Results— We optically mapped left ventricular wedge preparations from 12 failing native hearts and 2 rejected cardiac allografts explanted during transplant surgery. We determined transmural action potential duration (APD) before and after 100 nmol/L apamin administration in all wedges and after sequential administration of apamin, chromanol, and E4031 in 4 wedges. Apamin prolonged APD from 363 ms (95% confidence interval [CI], 341–385) to 409 (95% CI, 385–434; P P =0.001) in 12 native failing hearts at 1000 ms pacing cycle length (PCL). The percent APD prolongation is negatively correlated with baseline APD and positively correlated with PCL. Only 1 wedge had M-cell islands. The percentages of APD prolongation in the last 4 hearts at 2000 ms PCL after apamin, chromanol, and E4031 were 9.1% (95% CI, 3.9–14.2), 17.3% (95% CI, 3.1–31.5), and 35.9% (95% CI, 15.7–56.1), respectively. Immunohistochemical staining of subtype 2 of SK protein showed increased expression in intercalated discs of myocytes. Conclusions— SK current is important in the transmural repolarization in failing human ventricles. The magnitude of I KAS is positively correlated with the PCL, but negatively correlated with APD when PCL is fixed. There is abundant subtype 2 of SK protein in the intercalated discs of myocytes.

Objectives: We tested the acute effects of resynchronization in heart failure patients with a normal (>50%) left ventricular (LV) ejection fraction (HFNEF) and mechanical dyssynchrony. Methods: Twenty-four HFNEF patients (72 ± 6 years, 5 male) with mechanical dyssynchrony (standard deviation of electromechanical time delay among 12 LV segments >35 ms) were studied with temporary pacing catheters in the right atrium, LV, and right ventricle (RV), and high-fidelity catheters for pressure recording. Using selected atrioventricular (AV) intervals of 60, 90, 120, 150, and 180 ms to optimize transmitral flow during simultaneous biventricular pacing, the RV-LV (VV) interval was then evaluated at RV30, RV15, 0, LV15, LV30, and LV45 (RV or LV indicates which ventricle was paced first, the number indicates by how many ms). Results: During simultaneous pacing, longer AV intervals were associated with improved LV pressure-derivative minimums and increased aortic pressures (p < 0.05 vs. normal sinus rhythm). In the VV interval from RV30 to LV45, there was a graded increase in the aortic velocity time integral and a decrease in dyssynchrony during simultaneous or LV-first pacing (p < 0.05 vs. normal sinus rhythm). Conclusions: For HFNEF patients with mechanical dyssynchrony, acute simultaneous biventricular or LV-first pacing with longer AV intervals reduced mechanical dyssynchrony and improved diastolic and systolic hemodynamics.

The roundworm C. elegans is a mainstay of aging research due to its short lifespan and easily manipulable genetics. Current, widely used methods for long-term measurement of C. elegans are limited by low throughput and the difficulty of performing longitudinal monitoring of aging phenotypes. Here we describe the WorMotel, a microfabricated device for long-term cultivation and automated longitudinal imaging of large numbers of C. elegans confined to individual wells. Using the WorMotel, we find that short-lived and long-lived strains exhibit patterns of behavioral decline that do not temporally scale between individuals or populations, but rather resemble the shortest and longest lived individuals in a wild type population. We also find that behavioral trajectories of worms subject to oxidative stress resemble trajectories observed during aging. Our method is a powerful and scalable tool for analysis of C. elegans behavior and aging. DOI: http://dx.doi.org/10.7554/eLife.26652.001, eLife digest Aging affects almost all living things, yet little is known about the biological changes that occur as we get older. Scientists often study aging in the microscopic roundworm Caenorhabditis elegans because it reproduces quickly and its lifespan is short (about 2–3 weeks on average). To date, investigations have helped to reveal genes that affect overall lifespan. However, it is not known how much these genes also affect the animal’s healthy lifespan or “healthspan”, that is to say, the length of time before advancing age begins to negatively affect health. Until now, studies with worms have often been limited because measuring health and aging required time-consuming and difficult manual experiments. This also meant that worms were studied together as groups, rather than as individuals, providing a simplified picture of what was going on. An automated system in which many single worms can be analyzed and assessed would provide a much more detailed view of the effects of aging on health. Churgin et al. have now developed a device called the WorMotel to allow simultaneous automated examination of 240 worms throughout their entire adult lifespan. The WorMotel is a rectangular slab of clear silicone rubber with small wells in it. A single worm is confined in each well with a source of bacteria for food, and a camera is used to track and monitor each worm’s behavior over time. This device confirmed that worms move more slowly as they get older, which was taken to be a measurement of the worms’ declining health. Worms that lived the longest declined over the first few days and then had a long plateau of very low activity before eventually dying. Short-lived worms became slower and died fairly promptly. Churgin et al. also showed that the worms with mutations that increase lifespan declined in a similar way to the longest-lived normal worms, and that mutants with shorter lifespans declined like the shortest-lived normal worms. Also, normal worms that had been exposed to a chemical called paraquat – which stresses the worm's cells and shortens the worm’s lifespans to a few days – slowed down in a similar manner as aging worms, suggesting that the stress is similar to the aging process. Tools like the WorMotel can improve our understanding of the links between lifespan and healthspan. The tool is designed to be versatile and can be used with standard imaging systems and automated tools, meaning it can be scaled up to deal with tens of thousands of worms at once. Churgin et al. are now using the WorMotel to find other genes that influence healthspan and understand how they contribute to deteriorating health as animals age. Aging affects us all and learning more about healthspan could lead to drugs or interventions to help more people to live healthily for longer. DOI: http://dx.doi.org/10.7554/eLife.26652.002

The roundworm C. elegans is a mainstay of aging research due to its short lifespan and easily manipulable genetics. Current, widely used methods for long-term measurement of C. elegans are limited by low throughput and the difficulty of performing longitudinal monitoring of aging phenotypes. Here we describe the WorMotel, a microfabricated device for long-term cultivation and automated longitudinal imaging of large numbers of C. elegans confined to individual wells. Using the WorMotel, we find that short-lived and long-lived strains exhibit patterns of behavioral decline that do not temporally scale between individuals or populations, but rather resemble the shortest and longest lived individuals in a wild type population. We also find that behavioral trajectories of worms subject to oxidative stress resemble trajectories observed during aging. Our method is a powerful and scalable tool for analysis of C. elegans behavior and aging.

KRAS is a major oncogenic driver in pancreatic ductal adenocarcinoma (PDAC). Intriguingly, human tumors with RAS mutations often display loss of the remaining wild-type (WT) allele, including PDAC. We previously reported that selective loss of WT Kras allele is associated with PDAC progression to metastasis in both mice and humans. These data suggest that WT KRAS functions as a tumor suppressor in the context of mutant KRAS and loss of WT KRAS offers growth advantage in clonal evolution and tumor progression to metastasis. To investigate the underlying mechanism, we have examined the impact of inducible WT KRAS expression on human PDAC cell lines that had spontaneously lost the WT allele. We observed re-expression of WT KRAS significantly attenuated the malignancy of pancreatic cancer cells in vitro and in vivo. RNA-Seq and proteomic analyses identified HIPPO signaling, specifically YAP1 activation, is inhibited by WT KRAS restoration. We observed a marked reduction of YAP1 nuclear localization and a consequent inhibition of YAP1 transcriptional activity upon restoration of WT KRAS. Expression of constitutively activated YAP1 mitigated the growth-inhibitory effects of WT KRAS, confirming that YAP1 is a downstream effector of the pathway. Furthermore, YAP1 nuclear localization was found to be associated with poor prognosis in PDAC patients. Together, these results offer a novel mechanism that WT KRAS suppresses tumor metastasis via inhibition of YAP1 activation and further demonstrate that genomic loss of the WT KRAS allele is not a mere consequence of genomic instability but rather a selective event that promotes pancreatic tumor progression to metastasis. Citation Format: Han Yan, Chih-Chieh Yu, Stuart Fine, Ayman Lee, Ye-Ran Yang, Juan Yan, Dario Garcia-Carracedo, Dillon Karg, Edwin Cheung, Richard Friedman, Emily Chen, Ji Luo, Yi Miao, Wanglong Qiu, Gloria Su. Genomic loss of the wild-type KRAS allele in pancreatic tumor progression and metastasis [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr I17.

The electrical properties of Al2O3/n-InGaAs metal–oxide–semiconductor capacitors (MOSCAPs) with In content of 0.53, 0.7, and 1 (InAs) have been investigated. Results show small capacitance–voltage (C–V) frequency dispersion in accumulation (1.70% to 1.85% per decade) for these MOSCAPs, mostly being assigned to border traps in Al2O3. With higher In content, shorter minority-carrier response time and smaller C–V hysteresis are observed. The reduction of C–V hysteresis might be related to the reduction of Ga-bearing oxides in Al2O3/InGaAs interfaces as indicated by x-ray photoelectron spectroscopy.

Background The association between diastolic abnormality and postexercise contractile decompensation is uncertain in heart failure (HF) patients with reduced left ventricular ejection fraction (LVEF). Hypothesis The higher mitral E/annular early diastolic velocity (E/e′) is relevant to postexercise regional myocardial contractile maladaptation. Methods Seventy HF patients with LVEF 65 ms (OR: 64, 95% CI: = 6–651, P < 0.001) in HF patients with reduced LVEF in multivariate analysis. Conclusions The higher mitral E/e′-related postexercise maladaptation of myocardial contractile motion and synchronicity suggests the involvement of systolic abnormality in exercise pathophysiology in HF patients with reduced LVEF.

We investigated the measurement of soluble ST2 (sST2) in stable heart failure (HF) with a normal ejection fraction (HFNEF) in hypertensive patients.Echocardiography and serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) and sST2 concentrations were evaluated in 107 hypertensive patients (65 ± 12 years, 57 male) with ejection fraction (EF)50%. Among them, 68 patients with stable HF in functional class II and III were the HFNEF group. We found that the area under the receiver operating characteristic curve (AUC) for sST2 was 0.80 (95% CI 0.70-0.89; P.001), relatively better than that for NT-proBNP (AUC 0.70, 95% CI 0.58-0.79; P = .003) to detect HFNEF. However, the NT-proBNP concentration, rather than sST2, was higher in HFNEF patients with functional class III (562 ± 891 vs 185 ± 242 pg/mL in functional class II; P = .009), and correlated better with mitral E/e' (annular early diastolic velocity) (r = 0.327; P = .008) than sST2 concentrations in HFNEF patients. Multivariate analysis showed that sST213.5 ng/mL was independently associated with HFNEF in hypertensive patients (odds ratio 11.7, 95% CI = 2.9-47.4; P = .001).sST2 measurement provides diagnostic aid of stable HFNEF for hypertensive patients. Addition of NT-proBNP to sST2 could give further information regarding HF functional class and diastolic abnormality.

Background Apamin-sensitive small-conductance calcium-activated potassium (SK) channels are gated by intracellular Ca 2+ through a constitutive interaction with calmodulin. Objective We hypothesize that arrhythmogenic human calmodulin mutations impede activation of SK channels. Methods We studied 5 previously published calmodulin mutations (N54I, N98S, D96V, D130G, and F90L). Plasmids encoding either wild-type or mutant calmodulin were transiently transfected into human embryonic kidney 293 cells that stably express subtype 2 of SK protein channels (SK2 cells). Whole-cell voltage-clamp recording was used to determine apamin-sensitive current densities. We also performed optical mapping studies in normal murine hearts to determine the effects of apamin in hearts with (n=7) or without (n=3) pretreatment with sea anemone toxin. Results SK2 cells transfected with wild-type calmodulin exhibited an apamin-sensitive current density of 33.6 pA/pF (31.4–36.5 pA/pF) (median and confidence interval 25th–75th percentile), which was significantly higher than that observed for cells transfected with N54I (17.0 pA/pF [14.0–27.7 pA/pF]; P = .016), F90L (22.6 pA/pF [20.3–24.3 pA/pF]; P = .011), D96V (13.0 pA/pF [10.9–15.8 pA/pF]; P = .003), N98S (13.7 pA/pF [8.8–20.4 pA/pF]; P = .005), and D130G (17.6 pA/pF [13.8–24.6 pA/pF]; P = .003). The decrease in SK2 current densities was not associated with a decrease in membrane protein expression or intracellular distribution of the channel protein. Apamin increased the ventricular action potential duration at 80% repolarization (from 79.6 ms [63.4–93.3 ms] to 121.8 ms [97.9–127.2 ms]; P = .010) in hearts pretreated with anemone toxin but not in control hearts. Conclusion Human arrhythmogenic calmodulin mutations impede the activation of SK2 channels in human embryonic kidney 293 cells.

Atrial fibrillation (AF) is the most common sustained and most important arrhythmia in clinical practice. The mechanisms underlying AF initiation and maintenance are known to be complex and heterogeneous. The general understanding of the detailed molecular basis of AF is still incomplete. Recently, these is increasing evidence that small conductance calcium-activated potassium (SK) channels are associated with atrial action potential repolarization and the pathogenesis of AF. Although the functional role of SK channels in the genesis of AF is not entirely clear, new insights into the basic pathophysiological mechanism of AF have been provided. Besides, genome-wide association studies also implicate that genes coding for SK channels are related to the risk of developing AF. This article reviews recent work on the association of SK channels and AF, genetic studies of SK channels, and discuss future investigation and developments regarding this field.Atrial fibrillation; Genetics; Small conductance calcium-activated potassium channels.

Radiofrequency catheter ablation (RFCA) for atrial tachyarrhythmias in postoperative congenital heart disease (CHD) patients has a low success rate and a high recurrence rate. This study explores the reasons for these constraints.A total of 49 consecutive postoperative CHD patients who received RFCA for atrial tachyarrhythmias between 1993 and 2010 were enrolled.Overall, there were 86 RFCA procedures performed, 32 with the conventional method and 54 using CARTO-guided mapping. The interval between the operation and the first ablation was 13 years. Isthmus-dependent atrial flutter (AFL) was the most common type of tachycardia (37, 76%), followed by intra-atrial re-entry tachycardia (IART; 37%), and ectopic atrial tachycardia (EAT; 31%). By applying CARTO-guided mapping, the success rate was elevated compared to that of conventional ablation (84% vs. 56%, p = 0.006), but there was no improvement in the recurrence rate (22% vs. 28%, p = 0.75). Multiple atrial tachyarrhythmias occurred in 26 (53%) patients, and 17 presented during the initial electrophysiological study. The presence of multiple arrhythmias during the initial study predicted ablation failure or multiple ablations (11/17 vs. 3/32, p0.001). Among the 15 patients with new tachyarrhythmias, EAT and IART predominated. However, applying antiarrhythmia agents immediately following ablation may decrease arrhythmia recurrence (1/10 vs. 14/25, p = 0.02).Although electroanatomical mapping improves the results of RFCA in atrial tachyarrhythmias, the recurrence rate remains high because of multiple and new atrial tachyarrhythmias. Therefore, short-term pharmacological treatment following RFCA for positive remodeling should be considered.Ablation; Antiarrhythmia agents; Atrial tachyarrhythmia; Congenital heart disease; Multiple arrhythmias.

Fukuyama-type congenital muscular dystrophy (FCMD), the second most common childhood muscular dystrophy in Japan, is caused by alterations in the fukutin gene. Mutations in fukutin cause abnormal glycosylation of α-dystroglycan, a cell surface laminin receptor; however, the exact function and pathophysiological role of fukutin are unclear. Although the most prevalent mutation in Japan is a founder retrotransposal insertion, point mutations leading to abnormal glycosylation of α-dystroglycan have been reported, both in Japan and elsewhere. To understand better the molecular pathogenesis of fukutin-deficient muscular dystrophies, we constructed 13 disease-causing missense fukutin mutations and examined their pathological impact on cellular localization and α-dystroglycan glycosylation. When expressed in C2C12 myoblast cells, wild-type fukutin localizes to the Golgi apparatus, whereas the missense mutants A170E, H172R, H186R, and Y371C instead accumulated in the endoplasmic reticulum. Protein O-mannose β1,2-N-acetylglucosaminyltransferase 1 (POMGnT1) also mislocalizes when co-expressed with these missense mutants. The results of nocodazole and brefeldin A experiments suggested that these mutant proteins were not transported to the Golgi via the anterograde pathway. Furthermore, we found that low temperature culture or curcumin treatment corrected the subcellular location of these missense mutants. Expression studies using fukutin-null mouse embryonic stem cells showed that the activity responsible for generating the laminin-binding glycan of α-dystroglycan was retained in these mutants. Together, our results suggest that some disease-causing missense mutations cause abnormal folding and localization of fukutin protein, and therefore we propose that folding amelioration directed at correcting the cellular localization may provide a therapeutic benefit to glycosylation-deficient muscular dystrophies.

In this study, atomic layer deposition (ALD) has been applied to deposit aluminum doped zinc oxide (AZO) films as gas barriers, successfully. Diethylzinc and trimethylaluminum were used as precursors and water as oxidant. Sequential deposition steps of 400 ALD cycles have been adopted to prepare AZO films. Influences of deposition temperatures on electro-optical properties have studied by using Hall-effect measurement and transmittance spectrometer. AZO film deposited at 130 oC has a low resistivity of 5×10-3 Ω-cm. Water vapor transmittance rate has been carried out by using Mocon Permatran-W 3/61. The WVTR performances of ALD barriers, which highly related to the physical conditions of interfaces between nanolaminated structures, have been measured as 0.001 – 2 g/m2*25oC*day.

Recently zebrafish has become a powerful vertebrate model system for cardiac arrhythmia and another advantage is that the heart could be rapidly excised for in-vitro electrophysiological recording.We made direct in-vitro recordings of adult zebrafish heart ECG using the microelectrode and tested the effects of various drugs on zebrafish heart ECG.The QT interval change in the ECG was verified by optically mapped action potential duration (APD). The mean in-vitro heart rate (HR) of the adult zebrafish was 118 ± 22 beats/min. The mean QT interval was 312 ± 36 ms and mean HR corrected QT interval (QTc) 439 ± 39 ms. The mean optically mapped APD was 308 ± 30 ms, which was close to the QT interval in ECG (p=0.815). We found that sympathomimetic drug isoproterenol increased HR, whereas L-type calcium channel blocker verapamil decreased HR. Sodium channel blocker quinidine and L-type calcium channel activator BayK8644 prolonged QTc interval in a dose-dependent manner (515 ± 24 ms and 519 ± 27 ms, respectively, both p0.01). The APD was also prolonged accordingly. Both rapidly and slowly activating delayed rectifier potassium channel (IKr and IKs) blockers E4031 and chromanol 293B, respectively, also prolonged QTc interval in a dose-dependent manner (523 ± 25 ms and 529 ± 27 ms, respectively, both p0.01). Quinidine, E4031 and chromanol 293B also decreased HR.Direct in-vitro recording of adult zebrafish heart is an efficient platform for test of drugs which exert electrophysiological effects on cardiomyocytes, and perturbation of ionic channels that are responsible for human long QT syndrome also modulates QT interval in adult zebrafish heart.

Wing scales of male Euploea mulciber (E. mulciber) and Troides aeacus (T. aeacus) butterflies were investigated from interest in photonic crystal by scanning electron microscopy and optical reflectance measurement. On the basis of the structural observation, the colouration in different areas in their wings was discussed. It was particularly deduced that a violet-green iridescence characteristic of E. mulciber's forewing is caused only in a wavelength range from ∼380 to ∼510nm by multiple interference from a highly tilted, triple-layered cuticle arrangement on the brown scales. It was also found that T. aeacus does not produce a blue-green sheen such as observed by Troides magellanus because its scales have no multiple cuticle layers but microrib layers unable to produce any backscattering diffraction.

Considering full resolution and without 3D glasses, time-multiplexed type is the better choice for 3D display, such as directional sequential backlight system. However, the current directional sequential backlight system can only produce a single-user 3D display provided 3D image for single observer. Thus, the multi-user 3D film was proposed to add in front of the directional sequential backlight system and make the display become a multiuser (each observer watches the same image) 3D display provided full-resolution 3D images for three observers.

Marfan syndrome (MFS) is a rare disease that has a devastating outcome when cardiovascular complications occur. MFS diagnostic criteria have been revised several times since the causative mutation was detected. The most specific diagnostic phenotype is cardiovascular manifestation, which also signals the need for close follow-up; MFS patients may ultimately develop lethal complications during their third or fourth decade of life. An echocardiographer must be familiar with the cardiovascular presentation of the MFS patient and be aware of specific measures to be followed if MFS is suspected. Patients should be examined carefully for other disease symptoms to differentiate MFS from alternative diagnoses.

Background/Purpose This study examined the psychometric properties and clinical validity of the EORTC QLQ-PR25, a questionnaire for assessing the quality of life of patients with prostate cancer. Methods The Taiwan Chinese version of the prostate cancer module (EORTC QLQ-PR25) and the core questionnaires (EORTC QLQ-C30) were administered to 81 patients with prostate cancer after they had been treated with surgery or hormone therapy or both. The QLQ-PR25 module assesses urinary symptoms, bowel symptoms, hormonal treatment-related symptoms, sexual activity and sexual functioning. Results The questionnaires were well accepted by the patients and very few of the items had missing data. Only the urinary symptom scale showed satisfactory internal consistency. Scales were able to differentiate clinical groups of patients with corresponding symptoms, but the differences were smaller than that of major functioning scales in the core questionnaire. Conclusion The Taiwan Chinese version of the EORTC QLQ-PR25 is acceptable in patients with prostate cancer in Taiwan, able to differentiate corresponding symptoms, but the scale structure needs further improvement.

With atomic-layer-deposition grown zinc oxide as the electron selective layer, we developed plastic substrate compatible processing for organic photovoltaic devices and demonstrated flexible inverted organic solar cells on poly(ethylene naphthalate) with a power conversion efficiency of 4.18%.

The aim of this study was to establish a national database of mutations in the fibrillin-1 (FBN1) gene that cause Marfan syndrome (MFS) in the Taiwanese population. In this study, we screened 294 patients from 157 families for the presence of FBN1 mutations using polymerase chain reaction/ denaturing high performance liquid chromatography (PCR/DHPLC). We identified 56 mutations in 62 of the 157 (40%) families including 49 single-base substitutions (36 missense mutations, seven nonsense mutations, and six splicing sites), one small insertion, four small deletions, one small indel (insertion and deletion), and one exonic deletion (Exon 36). When family history was taken into consideration, the mutation detection rate rose to 91% (29 of 32). We further investigated the phenotypic data and found that one third (47 of 157) of the families fit the Ghent criteria for MFS. Based on that data, the mutation rate was 98% (46/47). That finding implies that family history and the Ghent criteria play a more important role than clinical manifestations in establishing a clinical diagnosis of Marfan syndrome. Among the 56 mutations found in this study, 40 (71%) have not been registered in the Human Gene Mutation Database (HGMD) or in the Universal Mutation Database (UMD). This is the first study of the mutation spectrum of MFS in a cohort of patients in Taiwan. The database is expected to considerably improve genetic counseling for and medical care of MFS families.

Solution cast and spin coating techniques were performed to deposit ferroelectric films of PVDF-PZT on Al 3+-ZnO/polyethylene terephthalate(PET) flexible substrates. The composites with 10wt% PZT in PVDF exhibited higher dielectric permittivity and showed better ferroelectric property, might be due to internal field generated by the nano PZT particles, which favours the orientation of PVDF molecules. Higher remanent polarization and low leakage current were successfully demonstrated by coating PVDF layer on PVDF-PZT composite film. The surface of the obtained PVDF films were smooth with less porosity above PVDF-PZT composite film, enabling a significant decrease in the leakage current of composite thin films.

Marfan syndrome has been associated with approximately 562 mutations in the fibrillin-1 ( FBN1 ) gene. Mutation scanning of the FBN1 gene with DNA direct sequencing is time-consuming and expensive because of its large size. This study analyzed the diagnostic value of high-resolution melting analysis as an alternative method for scanning of the FBN1 gene. A total of 75 polymerase chain reaction (PCR) amplicons (179–301 bp, average 256 bp) that covered the complete coding regions and splicing sites were evaluated on the 96-well LightCycler system. Melting curves were analyzed as fluorescence derivative plots (−d F /d T vs. temperature). To determine the sensitivity of this method, a total of 82 samples from patients with Marfan syndrome and 50 unaffected individuals were analyzed. All mutations reported in this study had been confirmed previously by direct sequencing analysis. Melting analysis identified 48 heterozygous variants. The variant c.3093 G>T (exon 25) was incorrectly identified by melting curve analysis. The sensitivity of the technique in this sample was 98.78% (81/82). This study demonstrated that high-resolution melting analysis is a reliable gene scanning method with greater speed than DNA sequencing. Our results support the use of this technology as an alternative method for the diagnosis of Marfan syndrome as well as its suitability for high-throughput mutation scanning of other large genes.

Pellets consisting of the para-selective HZSM-5 crystals prepared by the chemical liquid deposition (CLD) with tetraethyl orthosilicate (TEOS) were used as the catalyst for alkylation of toluene with ethylene in supercritical carbon dioxide. The pellets were formed by compressing the CLD modified HZSM-5 crystals with and without addition of the binder alumina. When the binder was used, the CLD was applied again right after the compression. For the addition of 0.1 ml of TEOS per gram of pellet, p -ethyltoluene yield and selectivity were found to be sufficiently close to those using the modified crystals and the compressed pellets without containing the binder. The alkylation was carried out in a tubular reactor packed with the prepared HZSM-5 pellets. From a study of the operating variables including temperature, pressure, weight hourly space velocity of toluene, and molar ratio of toluene to ethylene, the highest yield and selectivity of p -ethyltoluene were determined as 70 and 97%, respectively, at a temperature of 623 K, a pressure of 13.1 MPa, a weight hourly space velocity of 5.0 (g of toluene)/(g of catalyst)/h and a molar ratio of toluene to ethylene of 5. The obtained results indicate a pronounced shape-screening effect of the prepared HZSM-5 pellets and an applicability of using supercritical CO 2 as the carrier.

Low temperature processing of sol-gel spin coated ferroelectric Pb(Zr0.52Ti0.48)TiO3 (PZT) thin film and sputtered (La 0.7 Sr 0.3 )MnO 3 (LSMO) buffer layer on Cr/SiO2/Si and SUS430 substrates using a continuous wave CO2 laser annealing technique was studied in the present work. The laser annealed PZT films coated on LSMO/Cr/SiO2/Si substrates shows an enhancement in the remanent polarization from 6.22 to 11.94 μC/cm2 as the PZT irradiated with a laser fluence of 267 to 334 W/cm2. The Cr electrodes were oxidized easily which deteriorate the electrical properties of specimens, even upon using CO2 laser annealing. On the other hand, the LSMO films (0.2 μm) sputtered on SUS430 substrate and spin coated PZT thin films (0.2 μm) were laser annealed sequentially, showed remanent polarization from 18.07 to 24.01 μC/cm2. CO2 laser annealing not only improves ferroelectric properties and electric leakage currents of PZT films, but also diminishes the inter-diffusion of elements in electrode and ferroelectrics. More...

Pellets consisting of highly shape-selective HZSM-5 crystals prepared by chemical liquid deposition using tetraethyl orthosilicate (TEOS) as the silicon precursor were observed to be an effective c...

Permanent pacing is the treatment for chronotropically incompetent hearts. However, the right ventricular (RV) apical pacing-induced asynchrony, even maintaining the atrioventricular (AV) sequential activation, has depressed left ventricular contractility. Whether RV septal pacing would less compromise the electromechanical performance of the left ventricle and the chronotropic effect on myocardial contractility, is unknown.We prospectively studied 42 patients without structural heart diseases and with symptomatic bradycardia. There were 10 patients receiving atrial pacing (AAI) pacemakers, 18 patients having AV sequential pacing at RV apex (DDDapx) and 14 patients being AV sequentially paced at septum (DDDspt). Echocardiography was performed before and within 72 h after the pacemaker implantation. The ventricular mechanical performance and asynchrony was compared in conditions of programmed rates of 60, 80 and 100/min.Myocardial performance index was significantly better in DDDspt than in DDDapx patients (p=0.003). With faster programmed rate, the QRS/RR increased (p0.05) in DDDapx patients with more inter- and intraventricular asynchrony, implicating the disadvantage of prolonged depolarization time. The DDDspt group demonstrated comparable parameters of diastolic function to AAI patients and preserved mechanical performance during accelerated pacing.RV septal pacing showed the advantages of shorter depolarization time, less ventricular contractile asynchrony, better mechanical performance and preserved chronotropic response on myocardial contractility in comparison with apical pacing.

Aims While overexpression of TGF alpha has been reported in human pancreatic ductal adenocarcinoma (PDAC), mice with overexpressed TGF alpha develop premalignant pancreatic acinar-to-ductal metaplasia (ADM) but not PDAC. TGF-beta signaling pathway is pivotal to the development of PDAC and tissue fibrosis. Here we sought to investigate the interplay between TGFa and TGF-beta signaling in pancreatic tumorigenesis and fibrosis, namely via Smad4 inactivation. Methods The MT-TGF alpha mouse was crossed with a new Smad4 conditional knock-out mouse (Smad4(flox/flox); p48-Cre or S4) to generate Smad4(flox/flox); MT-TGF alpha; p48-Cre (STP). After TGFa overexpression was induced with zinc sulfate water for eight months, the pancreata of the STP, MT-TGF alpha, and S4 mice were examined for tumor development and fibrotic responses. PanIN lesions and number of ducts were counted, and proliferation was measured by Ki67 immunohistochemistry (IHC). Qualitative analysis of fibrosis was analyzed by Trichrome Masson and Sirius Red staining, while vimentin was used for quantification. Expression analyses of fibrosis, pancreatitis, or desmoplasia associated markers (alpha-SMA, Shh, COX-2, Muc6, Col1a1, and Ctgf) were performed by IHC and/or qRT-PCR. Results Our STP mice exhibited advanced ADM, increased fibrosis, increased numbers of PanIN lesions, overexpression of chronic pancreatitis-related marker Muc6, and elevated expression of desmoplasia-associated marker Col1A1, compared to the MT-TGFa mice. The inactivation of Smad4 in the exocrine compartment was responsible for both the enhanced PanIN formation and fibrosis in the pancreas. The phenotype of the STP mice represents a transient state from ADMs to PanINs, closely mimicking the interface area seen in human chronic pancreatitis associated with PDAC. Conclusion We have documented a novel mouse model, the STP mice, which displayed histologic presentations reminiscent to those of human chronic pancreatitis with signs of early tumorigenesis. The STP mice could be a suitable animal model for interrogating the transition of chronic pancreatitis to pancreatic cancer.

Parallel process units are often encountered in chemical process systems for different considerations, e.g., parallel reactors from reactor network synthesis, parallel columns for heat integration. One notable example is the feed split configuration of heatintegrated distillation columns. In this work, the control of parallel processes is illustrated with the heat-integrated distillation column example. Results show that more consistent control performance can be achieved using the proposed control structure for both feed composition and feed flow disturbances. More importantly, the improvement is obtained using simpler instrumentation and much less engineering effort.

Berberis sect. Wallichianae are species of evergreen shrubs that in Taiwan are found in subalpine and montane-temperate areas and which have a notoriously controversial taxonomic history. Based on multivariate statistical analyses of morphometric data and an explicitly stated species criterion, the taxonomy of the group in Taiwan and its close relative in Luzon ( B. barandana ) is revised and their endemicity is evaluated by molecular data. In addition to the six species recognized in the Flora of Taiwan, 2 nd ed. (i.e., B. aristatoserrulata , B. brevisepala , B. chingshuiensis , B. kawakamii , B. mingetsensis , and B. tarokoensis ), B. hayatana (synonymized under B. mingetsensis ) and B. nantoensis (synonymized under B. brevisepala ) are reinstated, and three new species ( B. pengii , B. ravenii , and B. schaaliae ) are described and illustrated. Phylogenetic analyses using three chloroplast DNA sequence regions ( rbcL , ycf6-psbM , and psbA-trnH ) place all Taiwanese species and B. barandana in a strongly supported clade derived from within the continental Asian species of sect. Wallichianae , indicating their independent evolutionary history and supporting their endemic status.

Brugada syndrome (BrS) is one of the ion channelopathies associated with sudden cardiac death (SCD). The most common BrS-associated gene (SCN5A) only accounts for approximately 20–25% of BrS patients. This study aims to identify novel mutations across human ion channels in non-familial BrS patients without SCN5A variants through disease-targeted sequencing. We performed disease-targeted multi-gene sequencing across 133 human ion channel genes and 12 reported BrS-associated genes in 15 unrelated, non-familial BrS patients without SCN5A variants. Candidate variants were validated by mass spectrometry and Sanger sequencing. Five de novo mutations were identified in four genes (SCNN1A, KCNJ16, KCNB2, and KCNT1) in three BrS patients (20%). Two of the three patients presented SCD and one had syncope. Interestingly, the two patients presented with SCD had compound mutations (SCNN1A:Arg350Gln and KCNB2:Glu522Lys; SCNN1A:Arg597* and KCNJ16:Ser261Gly). Importantly, two SCNN1A mutations were identified from different families. The KCNT1:Arg1106Gln mutation was identified in a patient with syncope. Bioinformatics algorithms predicted severe functional interruptions in these four mutation loci, suggesting their pivotal roles in BrS. This study identified four novel BrS-associated genes and indicated the effectiveness of this disease-targeted sequencing across ion channel genes for non-familial BrS patients without SCN5A variants.