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BACKGROUND: Vegetative insecticidal proteins (Vip3) from Bacillus thuringiensis (Bt) have been utilized for control of lepidopteran insect pests. The majority of known Vip3 proteins possess exceptional high toxicity against Noctuid insects such as the fall armyworm (FAW, Spodoptera frugiperda), beet armyworm (BAW, Spodoptera exigua) and cotton bollworm (CBW, Helicoverpa armigera), but generally have relatively low or even no activity against some very important pest insects, such as Asian corn borer (ACB, Ostrinia furnacalis), European corn borer (ECB, Ostrinia nubilalis), rice stem borer (RSB, Chilo suppressalis) and oriental armyworm (OAW, Mythimna separata). RESULTS: Here, we report mutant Vip3Af with a single amino acid mutation, Vip3Af-T686R, which gains significantly higher insecticidal activity against ACB, OAW and BAW, while retaining high activity against FAW, CBW and RSB. Protein proteolytic activation in vitro showed that the proteolytic activation efficiency of the mutant protein was greater than the wild-type protein in the midgut juice of ACB, OAW and BAW. Transgenic corn expressing this mutant Vip3Af showed high levels of resistance to ACB, OAW, FAW, BAW and CBW. CONCLUSION: Our results suggest that Vip3Af may be a superior Vip3A mutant for the development of transgenic crops with resistance to a broad range of lepidopteran pest species. © 2024 Society of Chemical Industry.

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Background Electrical cardioversion (EC) is a procedure that restores normal sinus rhythm in patients with atrial fibrillation (AF). Data on post-EC outcomes relative to the success of inpatient EC is limited. Methods This is a retrospective study of patients admitted for AF who underwent inpatient EC from January 1, 2017, to January 1, 2021. We collected demographics and clinical, biochemical, and echocardiographic parameters that impact the success of EC. Outcome events were 30-day readmissions and mortality. Results Our study included 54 unique patients who either had EC in the emergency room or as part of their hospital admission course for atrial fibrillation. Most patients were men with an average age of 70 years with traditional risk factors for cardiovascular disease including heart failure, coronary artery disease, and chronic kidney disease. The group who had unsuccessful cardioversion was older than those in the ineffective EC. Mortality at 30 days (p < 0.01), 1 year (p < 0.01), and 30-day readmission rate (p < 0.01) were higher in patients with unsuccessful EC. Conclusion A predictive model for successful EC remains difficult to establish. Patients with unsuccessful in-hospital EC are at high risk for mortality and readmission at 30 days and require a comprehensive pre-discharge multidisciplinary approach and prioritized and individualized post-discharge integrated care.

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A new and highly sensitive voltammetric technique was described in this study for the concurrent detection of endocrine disruptors bisphenol A (BPA) and bisphenol AF (BPAF) based on carbon nanocages (CNCs) and copper oxide nanochains (CuONCs). The CNCs was prepared by the solvothermal method and characterized using various techniques. Utilizing the nanocomposite of CNCs and CuONCs, the voltammetric sensor demonstrated outstanding performance in detecting BPA and BPAF simultaneously with distinct oxidation peaks and increased current peaks. The voltammetric signals have linear relationships with the two bisphenols ranging from 0.500 µM to 100 µM with a detection limit of 0.16 µM for BPA and 0.14 µM for BPAF. The newly designed sensor showed reliable consistency, long-term durability and anti-interference ability, and performed well in analyzing real water samples, indicating great potential for environmental monitoring.

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BACKGROUND: Preserving parathyroid function during thyroidectomy is crucial, but remains challenging. Real-time near-infrared autofluorescence (NIRAF) aids surgeons in intraoperative parathyroid gland (PTG) identification. However, its role in detecting PTGs unintentionally removed during surgery is unclear. STUDY DESIGN: This prospective study included adult patients undergoing endoscopic thyroidectomy. Surgeons identified and documented PTGs visually. Excised specimens underwent visual inspection and NIRAF imaging (PDE-Neo II). All fluorescent tissues were dissected and pathologically evaluated (reference standard). One scanned image per lobe was chosen to quantify autofluorescence (AF) intensity. RESULTS: Overall, 95 patients underwent endoscopic thyroidectomies, with NIRAF imaging applied to 152 excised lobes. Of these, 19 lobes displayed a total of 23 spots with increased intensity. 175 specimens were sent for pathological evaluation, and 7 were confirmed to be parathyroid tissue. NIRAF demonstrated 100.0 % sensitivity and 90.5 % specificity for predicting parathyroid tissue, with 30.4 % positive predictive value, 100.0 % negative predictive value of and 90.9 % accuracy. Quantitatively normalized, the AF signal intensity was significantly higher in NIRAF-positive tissues than negative (4.3 vs 1.2 times, p < 0.0001). Additionally, the AF signal intensity in regions pathologically confirmed of parathyroid tissue was higher than non-parathyroid tissue (9.1 vs 2.1 times, p < 0.0001). CONCLUSION: This study suggests that NIRAF has high sensitivity and specificity for detecting inadvertently resected PTGs after endoscopic thyroidectomy, contributing to preservation efforts. However, NIRAF-positive tissues still require additional confirmation through multiple methods, emphasizing other examinations to verify that they are indeed parathyroid tissues. Further research is warranted to refine NIRAF imaging parameters.

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Atrial fibrillation (AF) is a common cardiac arrhythmia that seriously affects the quality of life of patients. Effective treatment and prevention are important to control the morbidity and mortality of AF. It has been found that cardiac fibrosis promotes the onset and progression of AF. It is now known that transforming growth factor ß (TGF-ß), an important fibrotic cytokine, plays an important role in cardiac fibrosis by inducing myofibroblast activation via the activation of classical (SMAD-based) and non-classical (non-SMAD-based) signaling pathways. In addition, specific activation of the Wnt/ß-catenin pathway has been shown to promote the transformation of fibroblasts into myofibroblasts. In recent years, a new family of proteins, namely Disheveled-associated antagonist of beta-catenin (DACT) 2, can affect the Wnt/ß-catenin and TGF-ß signaling pathways by regulating the phosphorylation levels of these target proteins, which in turn affects the progression of fibrosis. The present study focuses on the effect of DACT2-guided ß-catenin on atrial fibrosis. It is expected that the summarized information can be helpful in the treatment of AF.

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Recently, there has been a major push toward the development of next-generation treatments against snakebite envenoming. However, unlike current antivenoms that rely on animal-derived polyclonal antibodies, most of these novel approaches are reliant on an in-depth understanding of the over 2000 known snake venom toxins. Indeed, by identifying similarities (i.e., conserved epitopes) across these different toxins, it is possible to design cross-reactive treatments, such as broadly-neutralising antibodies, that target these similarities. Therefore, in this project, we built an automated pipeline that generates sequence and structural distance matrices and homology trees across all available snake venom toxin sequences and structures. To facilitate analysis, we also developed a user-friendly and high-throughput visualisation tool, coined "Venom TOxin CluStering" (V-ToCs). This tool allows researchers to easily investigate sequence and structure patterns in snake venom toxins for a wide array of purposes, such as elucidating toxin evolution, and will also hopefully help guide the discovery and development of increasingly broadly-neutralising antivenoms in the near future.

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Recent artificial intelligence (AI) advancements in cardiovascular medicine offer potential enhancements in diagnosis, prediction, treatment, and outcomes. This article aims to provide a basic understanding of AI enabled ECG technology. Specific conditions and findings will be discussed, followed by reviewing associated terminology and methodology. In the appendix, definitions of AUC versus accuracy are explained. The application of deep learning models enables detecting diseases from normal electrocardiograms at accuracy not previously achieved by technology or human experts. Results with AI enabled ECG are encouraging as they considerably exceeded current screening models for specific conditions (i.e., atrial fibrillation, left ventricular dysfunction, aortic stenosis, and hypertrophic cardiomyopathy). This could potentially lead to a revitalization of the utilization of the ECG in the insurance domain. While we are embracing the findings with this rapidly evolving technology, but cautious optimism is still necessary at this point.

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The recently established non-thermal, single-shot pulsed field ablation (PFA) is a potential tool for achieving rapid pulmonary vein isolation (PVI) to cause cell death by electroporation, yet data regarding this state-of-the-art technology remain sparse. In this meta-analysis, we included 3,857 patients from 20 studies. There was no significant difference in AF recurrence between the PFA and control groups. Subgroup analysis showed that additional ablation beyond PVI has a similar rate of AF recurrence to PVI alone (10% versus 13%, respectively). PVI durability was achieved in 83% (mean), 95% CI [65-99%] of the PFA group and in 79% (mean), 95% CI [60-98%] of the control group, with no significant difference in the rate of PVI durability between the two groups. The PFA group had considerably reduced procedure duration, but not fluoroscopy time. No statistically significant differences in periprocedural complications were observed. PFA is associated with shorter procedural time than thermal ablation. Cardiac complications were uncommon and mainly reversible in both the PFA and control groups.

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As a replacement for bisphenol A (BPA), bisphenol AF (BPAF) showed stronger maternal transfer and higher fetal accumulation than BPA. Therefore, concerns should be raised about the health risks of maternal exposure to BPAF during gestation on the offspring. In this study, SD rats were exposed to BPAF (0, 50, and 100 mg/kg/day) during gestation to investigate the bioaccumulation and adverse effects in liver, spleen, and kidney tissues of the offspring at weaning period. Bioaccumulation of BPAF in these tissues with concentrations ranging from 1.56 ng/mg (in spleen of males) to 55.44 ng/mg (in liver of females) led to adverse effects at different biological levels, including increased relative weights of spleen and kidneys, histopathological damage in liver, spleen, and kidney, organ functional damage in liver, spleen, and kidney, upregulated expression of genes related to lipid metabolism (in liver), oxidative stress response (in kidney), immunity and inflammatory (in spleen). Furthermore, dysregulated metabolomics was identified in spleen, with 217 differential metabolites screened and 9 KEGG pathways significantly enriched. This study provides a comprehensive insight into the systemic toxicities of prenatal exposure to BPAF in SD rats. Given the broad applications and widespread occurrence of BPAF, its safety should be re-considered.

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Hepatocellular carcinoma, also referred to as HCC, is the most frequent form of primary liver cancer. It is anticipated that the discovery of the molecular pathways related with HCC would open up new possibilities for the treatment of HCC.WGCNA (Weighted gene co-expression network analysis) and molecular docking analysis were used to study the structural characteristics of POU2AF1 recombinant protein and its interaction with related proteins. Normal samples were placed in one group, and tumor samples were placed in another group inside the GEO database. We continued our investigation of the DEGs by performing an enrichment analysis using GO and KEGG. The GSCA platform is utilized in the process of doing an analysis of the connection between gene expression and medication sensitivity. In the end, the core target and the active molecule were both given the green light for a molecular docking investigation. POU2AF1 is being considered as a possible therapeutic target for HCC, and the results of our work have presented novel concepts for the treatment of HCC.

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BACKGROUND: No data have been reported on cooling characteristics and the impact of variant pulmonary vein (PV) anatomy on atrial fibrillation (AF) recurrences after POLARx cryoballoon (CB) ablation. OBJECTIVE: The purpose of this study was to analyze the impact of PV anatomy variants and cooling characteristics after CB ablation from a large multicenter prospective registry. METHODS: The primary end point was defined as 1-year absence of any atrial tachyarrhythmias (ATAs: AF/atrial flutter/atrial tachycardia). Correlation between ATA recurrences and anatomy variants/cooling characteristics were evaluated. The secondary outcome was the rate of major periprocedural complications. RESULTS: A total of 429 consecutive patients diagnosed with paroxysmal AF (83.4%) or persistent AF (peAF; 16.6%) were enrolled. Twenty-eight patients (6.6%) exhibited an anatomical variant (common ostium: 4.0%; adjunctive PV: 2.6%). Nadir temperature, thaw time, and total deflation time were different between standard PVs and PV variants. After the blanking period, over a mean of 431 ± 99 days of follow-up, 63 patients (14.7%) suffered an ATA recurrence. Patients with recurrences had both a shorter thaw time (18.5 ± 7 seconds vs 19.8 ± 7 seconds; P = .0012) and a shorter total deflation time, whereas time to isolation was longer (57.4 ± 42 seconds vs 49.1 ± 33 seconds; P = .04). Patients with anatomy variants showed a similar ATA recurrence rate (5 of 28 [17.9%]) to the standard anatomy group (58 of 401 [14.5%]) (P = .584), with a hazard ratio (HR) of 1.43 (95% confidence interval [CI] 0.49-4.13; log-rank, P = .4384). After adjusting for confounders, heart failure (HR 4.12; 95% CI 1.75-9.73; P = .0013) and peAF (HR 1.81; 95% CI 1.03-3.18; P = .0433) remained associated with ATA recurrence during follow-up. CONCLUSION: The POLARx CB system demonstrated long-term efficacy, along with a safe profile, in both patients with paroxysmal AF and those with peAF, regardless of the presence PV variants. Time to isolation was longer in patients with ATA recurrences during follow-up. CLINICAL TRIAL REGISTRATION: Catheter Ablation of Arrhythmias with a High-Density Mapping System in Real-World Practice (CHARISMA). CLINICALTRIALS: gov identifier: NCT03793998. Registration date: January 4, 2019.

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Atrial fibrillation (AF) is a prevalent cardiac arrhythmia associated with an increased risk of stroke due to disrupted heart function and potential clot formation. This review examines current management strategies for stroke prevention in AF, focusing on the efficacy, safety, and long-term outcomes of anticoagulation therapies. Anticoagulants, including novel oral anticoagulants (NOACs) and vitamin K antagonists, play a crucial role in reducing stroke risk by preventing clot formation in the heart. Recent studies highlight NOACs as superior alternatives to traditional therapies, offering improved safety profiles and enhanced patient adherence. Despite the risk of bleeding complications, judicious use of anticoagulants significantly improves clinical outcomes in AF patients. The review synthesizes evidence from clinical trials and meta-analyses to underscore the pivotal role of NOACs in transforming stroke prevention strategies in AF. Moreover, it discusses emerging interventions such as left atrial appendage occlusion and emphasizes the importance of personalized, patient-centered care in optimizing treatment decisions for AF patients at risk of stroke.

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The influence of air pollution on dynamic changes in clinical state from healthy to atrial fibrillation (AF), further AF-related complications and ultimately, death are unclear. We aimed to investigate the relationships between air pollution and the occurrence and progression trajectories of AF. We retrieved 442,150 participants free of heart failure (HF), myocardial infarction (MI), stroke and dementia at baseline from UK Biobank. Exposures to air pollution for each transition stage were estimated at the geocoded residential address of each participant using the bilinear interpolation approach. The outcomes were incident AF, complications, and death. Multi-stage models were used to evaluate the associations between air pollution and dynamic progression of AF. Over a 12.6-year median follow-up, a total of 21,670 incident AF patients were identified, of whom, 4103 developed complications and 1331 died. PM2.5, PM10, NOx and NO2 were differentially positively associated, while O3 was negatively associated with risks of progression trajectories of AF. PM2.5 exposure was significantly associated with an increased risk of progression. The associations of PM2.5, PM10, NOx, and NO2 on incident AF were generally more pronounced compared to other transitions. The cumulative transition probabilities were generally higher in individuals with higher exposure levels of PM2.5, PM10, NOx, and NO2 and lower exposure to O3. Air pollution could potentially have a role in increasing the risk of both the occurrence and progression of AF, emphasizing the significance of air pollution interventions in both the primary prevention of AF and the management of AF-related outcomes.

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BACKGROUND: Atrial fibrosis has a significant impact on the success rate of catheter ablation (CA) treatment of atrial fibrillation (AF). The fibrotic tissues could be reflected by the amplitude of the fibrillatory wave (F-wave). METHODS AND RESULTS: 704 patients with persistent AF and at least 1-year follow-up after CA were included as the internal group. 101 patients from another hospital were used as the external validation cohort. A 12­lead ECG was performed before CA and the maximum FWA in three ECG leads (aVL, aVF, V1) were measured. The FWA score (0 to 6 points according to the amplitude range of the three leads) of each patients was calculated. Five models including clinical features, FWA score, CHA2DS2-VASc score, APPLE score and the fusion of clinical features and FWA score were built. The FWA score was superior to the model constructed by clinical variables, CHA2DS2-VASc score and APPLE score. It not only had good predictive performance for AF recurrence, with an AUC value of 0.812 (95% CI 0.724-0.900), but also showed a significant predictive value for the recurrence rate according to F-wave amplitude. In the external validation cohort, the FWA score showed similar results (AUC 0.768, 95% CI 0.672-0.865). CONCLUSIONS: The present study reveals the significant predictive value of the FWA score for persistent AF ablation recurrence.

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Somatic mutations in genes encoding components of the RNA splicing machinery occur frequently in multiple forms of cancer. The most frequently mutated RNA splicing factors in cancer impact intronic branch site and 3' splice site recognition. These include mutations in the core RNA splicing factor SF3B1 as well as mutations in the U2AF1/2 heterodimeric complex, which recruits the SF3b complex to the 3' splice site. Additionally, mutations in splicing regulatory proteins SRSF2 and RBM10 are frequent in cancer, and there has been a recent suggestion that variant forms of small nuclear RNAs (snRNAs) may contribute to splicing dysregulation in cancer. Here, we describe molecular mechanisms by which mutations in these factors alter splice site recognition and how studies of this process have yielded new insights into cancer pathogenesis and the molecular regulation of splicing. We also discuss data linking mutant RNA splicing factors to RNA metabolism beyond splicing.

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BACKGROUND: Microsatellite instability (MSI) due to mismatch repair deficiency (dMMR) is common in colorectal cancer (CRC). These cancers are associated with somatic coding events, but the noncoding pathophysiological impact of this genomic instability is yet poorly understood. Here, we perform an analysis of coding and noncoding MSI events at the different steps of colorectal tumorigenesis using whole exome sequencing and search for associated splicing events via RNA sequencing at the bulk-tumor and single-cell levels. RESULTS: Our results demonstrate that MSI leads to hundreds of noncoding DNA mutations, notably at polypyrimidine U2AF RNA-binding sites which are endowed with cis-activity in splicing, while higher frequency of exon skipping events are observed in the mRNAs of MSI compared to non-MSI CRC. At the DNA level, these noncoding MSI mutations occur very early prior to cell transformation in the dMMR colonic crypt, accounting for only a fraction of the exon skipping in MSI CRC. At the RNA level, the aberrant exon skipping signature is likely to impair colonic cell differentiation in MSI CRC affecting the expression of alternative exons encoding protein isoforms governing cell fate, while also targeting constitutive exons, making dMMR cells immunogenic in early stage before the onset of coding mutations. This signature is characterized by its similarity to the oncogenic U2AF1-S34F splicing mutation observed in several other non-MSI cancer. CONCLUSIONS: Overall, these findings provide evidence that a very early RNA splicing signature partly driven by MSI impairs cell differentiation and promotes MSI CRC initiation, far before coding mutations which accumulate later during MSI tumorigenesis.

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INTRODUCTION: Strategies beyond pulmonary vein isolation (PVI) in persistent atrial fibrillation (persAF) are debated. A novel mapping tool provides algorithmic detection of ablation targets based on electrogram (EGM) properties specific to stable localized rotational activations. METHODS: The mapping tool was used on 31 patients (20 de novo). The algorithm was used to optimize PVI line placement and guide additional ablations. Targets were detected by calculating local cycle length (L-CL) and local spread of activation within that L-CL (Duty Cycle; DC) for EGMs with consistent morphology and activation. At least two left atrial (LA) maps (pre-PVI and post-PVI) were acquired in atrial fibrillation (AF) in all patients (except those with AF termination during PVI). Extra-pulmonary vein (PV) targets were compared between the two LA maps in each patient. Follow-up included Holter monitoring every 3 months. RESULTS: Patients had a median of 3 extra-PV drivers/targets. The majority (81%) were localized in the same areas between the two LA maps. All patients had progressive AF organization demonstrated by global activation slowing: histogram peak L-CL increased from 162 to 171 ms (post-PVI; p = .0003) than to 175 ms (posttarget ablation; p = .04). Moreover, L-CL dispersion was reduced by ablation; in 50% their values tended to cluster around two dominant cycles. In de novo patients AF terminated to sinus rhythm or atrial tachycardia (AT) within 48 h postprocedure in 88% of cases, and at 18 months mean follow-up recurrence occurred in only five (25%) patients (three persAF, two AT). There were no complications. CONCLUSION: The algorithmic detection of EGMs consistent with localized reentry during sequential mapping of persAF provided reproducible targets for ablation. This allowed personalized PVI and limited, highly-selective, extra-PV ablation. Results of this initial experience included progressive organization of AF with ablation and a low recurrence rate after a single procedure.

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OBJECTIVE: The benefits of rhythm control in early atrial fibrillation (AF) are increasingly recognized. This study aimed to investigate whether early AF ablation contributes to long-term sinus rhythm maintenance and to identify a suitable predictive score. METHODS: According to diagnosis-to-ablation time, this study prospectively enrolled 245 patients with very early AF, 262 with early AF, and 588 with late AF for radiofrequency ablation from June 2017 to December 2022. Clinical data, risk scores, and follow-up results were collected and analyzed. RESULTS: Baseline characteristics were similar among the three cohorts. During a median follow-up period of 26 months, AF recurrence was observed in 61 (24.9%), 66 (25.2%), and 216 (36.7%) patients in the very early, early, and late AF cohorts, respectively. In the multivariable-adjusted model, very early and early AF were associated with a reduced risk of AF recurrence, with hazard ratios of 0.72 (95% confidence interval [CI] 0.52-0.99) and 0.57 (95% CI 0.41-0.78), respectively. The APPLE score demonstrated the highest predictive power for very early AF, with an area under the curve (AUC) of 0.74. However, its predictive power decreased with time from diagnosis, showing low predictive power for late AF (AUC = 0.58). In addition, the time-dependent concordance index showed consistent results. For very early AF, the Akaike information criterion and decision curve analysis showed that APPLE had the highest predictive value. CONCLUSION: Very early AF ablation was associated with a lower recurrence rate, and the APPLE score provided a higher predictive value for these patients. (URL: https://www.chictr.org.cn/; Unique identifier: ChiCTR-OIN-17013021).

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Introduction: Rotary systems have made significant advances to improve their root canal preparation efficacy. These instruments can properly preserve the root canal anatomy and morphology. The present in vitro study aimed to compare canal transportation and centering ability of RaCe and Af f-one systems using cone-beam computed tomography. Materials and Methods: Thirty-six mandibular molars were included. The samples were randomly assigned to two groups (n=18): group 1, RaCe, and group 2, Af f-one. Canal preparation was conducted using the respective files according to the manufacturers' instructions. The cone-beam computed tomographic scanning of the samples was performed before and after preparation. The data were analyzed by using two-way ANOVA. Results: In both the RaCe and Af f-one rotary systems, canal centrality and transportation were similar at coronal, middle, and apical cross-sections. In addition, canal transportation and centrality were identical in the RaCe and Af f-one rotary files (P<0.05). Conclusion: The two studied rotary systems did not exhibit significant differences in root canal transportation and preservation of root canal centrality in the apical, middle, and coronal thirds.

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Background: Observational studies have shown that heart rate (HR), heart rate variability (HRV), P-wave terminal force, P-wave duration, T-wave amplitude and PR interval are associated with risk factors for atrial fibrillation (AF) or bradycardia. Arrhythmias are associated with many causes of hospitalization. However, observational studies are susceptible to confounding factors that have not yet been identified. The objective of this study was to clarify the causal relationships by Mendelian randomization analysis. Methods: We conducted a two-sample and multivariate Mendelian randomization (MVMR) analysis using genome-wide association study (GWAS) data from a European population to assess the total and direct causal effects of HR, three HRV traits, P-wave terminal force, P-wave duration, T-wave top amplitude in five-lead modes, and the PR interval on the risk of AF (N=191,205), bradycardia (N=463,010), and supraventricular tachycardia (SVT) (N=463,010). Results: The results of the univariate MR analysis revealed the following significant causal effects: the higher the genetically predicted PR interval, the lower the risk of AF; the higher the HR and T-wave top amplitude (aVR leads and V3 + V4 + aVL leads), the lower the risk of bradycardia; and the higher HR and the lower PR interval, the higher the risk of SVT. The multivariate MR results indicated that the HRV_standard deviation of the normal-to-normal (SDNN) interval had an independent causal effect on the risk of AF [odds ratio (OR): 0.515; 95% confidence interval (CI): 0.278-0.954; P=0.03], and the T-wave top amplitude in the aVR leads (OR: 0.998; 95% CI: 0.996-0.999; P<0.001) and the HRV_SDNN (OR: 0.988; 95% CI: 0.976-1.000; P=0.045) had independent causal effects on the risk of bradycardia. Conclusions: The HRV_SDNN had an independent causal effect on AF, while the HRV_SDNN and T-wave top amplitude in the aVR leads had independent causal effects on bradycardia, which suggests that some of the electrocardiographic parameters have preventive effects on the incidence of AF and bradycardia.