Search

RNA-binding proteins (RBPs) modulate alternative splicing outcomes to determine isoform expression and cellular survival. To identify RBPs that directly drive alternative exon inclusion, we developed tethered function luciferase-based splicing reporters that provide rapid, scalable and robust readouts of exon inclusion changes and used these to evaluate 718 human RBPs. We performed enhanced cross-linking immunoprecipitation, RNA sequencing and affinity purification-mass spectrometry to investigate a subset of candidates with no prior association with splicing. Integrative analysis of these assays indicates surprising roles for TRNAU1AP, SCAF8 and RTCA in the modulation of hundreds of endogenous splicing events. We also leveraged our tethering assays and top candidates to identify potent and compact exon inclusion activation domains for splicing modulation applications. Using these identified domains, we engineered programmable fusion proteins that outperform current artificial splicing factors at manipulating inclusion of reporter and endogenous exons. This tethering approach characterizes the ability of RBPs to induce exon inclusion and yields new molecular parts for programmable splicing control.

RNA binding proteins (RBPs) are key regulators of RNA processing and cellular function. Technologies to discover RNA targets of RBPs such as TRIBE (targets of RNA binding proteins identified by editing) and STAMP (surveying targets by APOBEC1 mediated profiling) utilize fusions of RNA base-editors (rBEs) to RBPs to circumvent the limitations of immunoprecipitation (CLIP)-based methods that require enzymatic digestion and large amounts of input material. To broaden the repertoire of rBEs suitable for editing-based RBP-RNA interaction studies, we have devised experimental and computational assays in a framework called PRINTER (protein-RNA interaction-based triaging of enzymes that edit RNA) to assess over thirty A-to-I and C-to-U rBEs, allowing us to identify rBEs that expand the characterization of binding patterns for both sequence-specific and broad-binding RBPs. We also propose specific rBEs suitable for dual-RBP applications. We show that the choice between single or multiple rBEs to fuse with a given RBP or pair of RBPs hinges on the editing biases of the rBEs and the binding preferences of the RBPs themselves. We believe our study streamlines and enhances the selection of rBEs for the next generation of RBP-RNA target discovery.

Abstract Background Catheter ablation in the pediatric population using fluoroscopy has been known to cause adverse events. This study aims to assess the effectiveness and safety of zero fluoroscopy (ZF) and near‐ZF‐guided catheter ablation for the treatment of arrhythmias in the pediatric population. Methods The PubMed, Embase, and Cochrane library databases were searched and reviewed for relevant studies. Outcomes of interest include safety, short‐term, and long‐term effectiveness. We classified patients ≤21 years old who underwent ZF or near‐ZF ablation with fluoroscopy time ≤1.5 min as our study group and patients within the same age range who underwent conventional fluoroscopy and/or near‐ZF ablation with a mean fluoroscopy time >1.5 min as our control group. Both ZF and near‐ZF ablation utilized 3D‐electroanatomical mapping (3D‐EAM). Results Ten studies composed of 2279 patients were included in this study. Total fluoroscopy time (MD –15.93 min, 95% CI (−22.57 – (−9.29), p

Abstract Background Ventricular tachycardia (VT) remains a risk in repaired Tetralogy of Fallot (rTOF); however, long‐term benefits of VT ablation have not been established. This study compares the outcomes of rTOF patients with and without VT ablation. Methods We searched multiple databases examining the outcomes of rTOF patients who had undergone VT ablation compared to those without ablation. Primary outcomes were VT recurrence, sudden cardiac death (SCD), and all‐cause mortality. Subgroup analysis was conducted based on the type of ablation (catheter and surgical). Slow‐conducting anatomical isthmus (SCAI)‐based catheter ablation (CA) was also analyzed separately. The secondary outcome was the risk factors for the pre‐ablation history of VT. Results Fifteen cohort studies with 1459 patients were included, 21.4% exhibited VTs. SCAI was found in 30.4% of the population, with 3.7% of non‐inducible VT. Factors significantly associated with VT before ablation included a history of ventriculostomy, QRS duration ≥180 ms, fragmented QRS, moderate to severe pulmonary regurgitation, high premature ventricular contractions burden, late gadolinium enhancement, and SCAI. Ablation was only beneficial in reducing VTs recurrence in SCAI‐based CA (risk ratio (RR) 0.11; 95% CI 0.03 to 0.33. p

HIGHLIGHTS Younger age, black race, normotension, and multiparity indicate a poorer prognosis for peripartum cardiomyopathy recovery, while bromocriptine therapy reduces adverse events. ABSTRACT Objectives: This study aimed to fill the significant knowledge gap regarding peripartum cardiomyopathy (PPCM), a heart failure phenotype linked to pregnancy. The main objectives were to explore the factors influencing the development and progression of PPCM and to assess the outcomes of bromocriptine. Materials and Methods: Systematic search across PubMed, ScienceDirect, and Cochrane Library identified studies until December 2022. This study includes non-randomized prospective and retrospective studies, as well as relevant randomized controlled trials. Risk factors were compared between the recovered and non-recovered PPCM groups, and bromocriptine therapy outcomes were evaluated against standard heart failure treatment as the primary endpoint. Results: The analysis included 24 observational studies and 1 randomized controlled trial involving 1,651 PPCM patients; 9 studies evaluating the outcomes of bromocriptine therapy. The most prevalent factors were caesarean delivery (proportion=53%, 95%CI=41%-66%) and anemia (proportion=51%, 95%CI=38%-65%). Non-recovered patients were younger (MD=-1.04 years old, 95%CI=-1.82-(-0.27), p=0.008) and predominantly black (RR=1.82, 95%CI=1.43-2.31, p

Background: The geographical diversity and clinical presentation differences of acute coronary syndrome (ACS) patients in Bali significantly influence the management performance and intervention outcomes. Bali, an island with varied geography, presents unique challenges for timely access to percutaneous coronary intervention (PCI) facilities, which are concentrated in South Bali. This study aims to describe annually the clinical characteristics of ACS patients in Bali, focusing on geographical factors and management performance. Methods: The regional data from the Indonesia Acute Coronary Syndrome Registry (One ACS) were utilized in this study. The study population consisted of patients aged over 18 years who were treated for ACS from 2017 to 2023 at Prof. Dr. I.G.N.G Ngoerah Central General Hospital in Denpasar, Bali. Several outcomes were described, including baseline characteristics, transfer time, door-to-balloon time (DTB), door-to-wire time (DTW), ambulance transport time, patient transport time, and major adverse cardiac event (MACE) outcomes. Results: A total of 4588 ACS patients based in Bali were included from 2017 to 2023. The majority of ACS patients originated from Denpasar city (44.1 %), followed by Badung (19.2 %) and Gianyar (8.5 %). More than half of the patients (57.9 %) were referred from primary healthcare, with a mean transfer time of 247.45 ± 698.26 min. Mortality rates were high, with significant numbers of cardiac arrests, heart failure, and cardiogenic shock. On multivariate analysis, shortness of breath, chest pain during light activity, patients seeking first medical contact (FMC) in Karangasem, and self-admission to the hospital were independently associated with mortality. Conclusions: The geographical challenges of Bali, particularly the limited distribution of PCI-capable facilities, contribute to delays in treatment and varied outcomes for ACS patients. Improvements in healthcare systems across Bali are required to conform with current guidelines and enhance intervention outcomes.

Abstract RNA binding proteins (RBPs) are key regulators of RNA processing and cellular function. Technologies to discover RNA targets of RBPs such as TRIBE (targets of RNA binding proteins identified by editing) and STAMP (surveying targets by APOBEC1 mediated profiling) utilize fusions of RNA base-editors (rBEs) to RBPs to circumvent the limitations of immunoprecipitation (CLIP)-based methods that require enzymatic digestion and large amounts of input material. To broaden the repertoire of rBEs suitable for editing-based RBP-RNA interaction studies, we have devised experimental and computational assays in a framework called PRINTER (protein-RNA interaction-based triaging of enzymes that edit RNA) to assess over thirty A-to-I and C-to-U rBEs, allowing us to identify rBEs that expand the characterization of binding patterns for both sequence-specific and broad-binding RBPs. We also propose specific rBEs suitable for dual-RBP applications. We show that the choice between single or multiple rBEs to fuse with a given RBP or pair of RBPs hinges on the editing biases of the rBEs and the binding preferences of the RBPs themselves. We believe our study streamlines and enhances the selection of rBEs for the next generation of RBP-RNA target discovery.

Abstract Background Inherited Primary Arrhythmias Syndromes (IPAS), especially Brugada syndrome (BrS), have been associated with arrhythmogenic substrates that can be targeted through ablation. This meta‐analysis evaluated the outcomes of catheter ablation (CA) in different types of IPAS based on procedural guidance and location. Methods A systematic search was conducted across multiple databases to identify studies reporting on ventricular arrhythmia (VA) events before and after CA in IPAS, including BrS, Long‐QT syndrome (LQTS), Early repolarization syndrome (ERS), and Idiopathic ventricular fibrillation (IVF). The primary outcomes were VA recurrence and VA burden, evaluated through conditional subgroup analysis. Procedural data were collected as secondary outcomes. Results A total of 21 studies involving 584 IPAS patients who underwent CA were included. Following a mean follow‐up duration of 33.5 months, substrate‐based ablation demonstrated efficacy in reducing VA recurrence across all types of IPAS [RR 0.23; 95% CI (0.13–0.39); p

Background: Ejection fraction (EF) is often used as a prognostic indicator and for classifying heart failure (HF) patients. This study evaluates the association of echocardiographic parameters with HF with improved EF (HFimpEF). Methods: This single-centre study retrospectively included patients with HF with reduced EF (HFrEF) from a cohort of admitted patients over 2018-2020, who were then followed up prospectively until 2023. The control group was categorized as patients with non-recovered HFrEF, and the population group was categorized as patients with HFimpEF. Results: A total of 176 patients with HFrEF were included in the study. Non-ischemic etiology was found to be the most prevalent cause of HFimpEF. The baseline echocardiography examination revealed that the HFimpEF group exhibited significantly higher values for tricuspid annular plane systolic excursion (TAPSE; P < 0.001) and inferior vena cava diameter (P < 0.001). The non-recovered HFrEF group demonstrated higher baseline left atrial volume index (LAVi) values (P < 0.001). In multivariate analysis, a higher value of TAPSE (odds ratio 3.071; P = 0.008) and a lower value of LAVi (odds ratio 2.034; P = 0.008) were independent echocardiography variables associated with HFimpEF. After a mean follow-up duration of 32.5 ± 9.1 months, the HFimpEF group had higher survival from rehospitalization due to worsening HF and lower all-cause mortality (log rank P < 0.001 and P = 0.005, respectively). Conclusions: Higher TAPSE and lower LAVi in baseline were associated with the transition from HFrEF to HFimpEF. The HFimpEF group had better survival compared to those with non-recovered HFrEF. Résumé: Contexte: La fraction d’éjection est souvent utilisée comme indicateur pronostique et comme élément de classification des patients atteints d’insuffisance cardiaque. La présente étude visait à évaluer l’association entre les paramètres échocardiographiques et l’insuffisance cardiaque avec fraction d’éjection améliorée (ICFEA). Méthodologie: Cette étude monocentrique a été menée de façon rétrospective auprès d’une cohorte de patients atteints d’insuffisance cardiaque avec fraction d’éjection réduite (ICFER) traités entre 2018 et 2020, et cette cohorte a été suivie de façon prospective jusqu’en 2023. Les patients du groupe témoin ont été classés comme ayant une ICFER ne s’étant pas résorbée, et les patients de la population étudiée ont été classés comme ayant une ICFEA. Résultats: Au total, 176 patients présentant une ICFER ont été inclus dans l’étude. La cause la plus fréquente d’ICFER était une étiologie non ischémique. Lors de l’évaluation échocardiographique initiale, les patients du groupe ayant progressé vers l’ICFEA présentaient des valeurs significativement plus élevées en ce qui concerne l’excursion systolique du plan de l’anneau tricuspide (TAPSE pour tricuspid annular plane systolic excursion) (p < 0,001) et le diamètre de la veine cave inférieure (VCI) (p < 0,001). D’autre part, les patients du groupe dont l’ICFER ne s’est pas résorbée présentaient des valeurs initiales plus élevées à l’indice de volume auriculaire gauche (IVAG) (p < 0,001). Lors d’une analyse multivariée, des valeurs de TAPSE plus élevées (rapport de cotes [RC] de 3,071; p = 0,008) et des valeurs plus faibles d’IVAG (RC de 2,034; p = 0,008) étaient deux variables échocardiographiques indépendantes associées avec la progression vers l’ICFEA. Après un suivi d’une durée moyenne de 32,5 ± 9,1 mois, le groupe présentant une ICFEA présentait un taux plus élevé de survie sans réhospitalisation due à une aggravation de l’IC et un taux plus faible de mortalités toutes causes confondues que le groupe dont l’ICFER ne s’était pas résorbée (p selon le test logarithmique par rangs < 0,001 et p = 0,005, respectivement). Conclusions: Une valeur de TAPSE élevée et un IVAG faible à l'évaluation initiale étaient associés à un passage de l’ICFER à l’ICFEA. La survie de patients présentant une ICFEA était supérieure à celle des patients présentant une ICFER non résorbée.

Abstract Background Ruptured sinus of Valsalva aneurysm (RSoVA) is an uncommon cardiac anomaly that may occasionally coexist with additional congenital heart disease (CHD). The presence of such congenital cardiac anomalies, frequently involving a ventricular septal defect (VSD), is of significant clinical importance and warrants vigilant detection during echocardiographic assessments to prevent oversight. Case presentation Three cases of RSoVA accompanied by VSD are presented in which all patients manifested symptoms of shortness of breath (SOB). In the first patient, right and left heart catheterization was undertaken; however, the images failed to reveal any evidence of VSD flow due to the occlusion of a small VSD by the prolapsed right coronary cusp (RCC). Prior to surgical intervention, multimodal imaging was conducted, revealing the presence of RSoVA extending into the right ventricle (RV) along with the VSD. The second patient had a prior childhood diagnosis of VSD but had not undergone further medical intervention. Transoesophageal echocardiography (TEE) was subsequently performed, identifying the presence of RSoVA, a small VSD, and valvular heart disease. The third patient presented with heart failure, exacerbated during her second pregnancy. TEE was also administered to this patient, revealing the presence of RSoVA accompanied by a small subaortic VSD with a left-to-right shunt. All three patients were scheduled for surgical repair of the ruptured sinus of Valsalva and closure of the VSD. Conclusions The coexistence of RSoVA and CHD, typically VSD, is frequently observed in patients experiencing symptoms of SOB. Recognizing the presence of CHD in individuals with RSoVA is of paramount importance, as it can significantly influence their medical management and treatment strategies.

Determining the layers of gene regulation within the innate immune response is critical to our understanding of the cellular responses to infection and dysregulation in disease. We identified a conserved mechanism of gene regulation in human and mouse via changes in alternative first exon (AFE) usage following inflammation, resulting in changes to the isoforms produced. Of these AFE events, we identified 95 unannotated transcription start sites in mice using a de novo transcriptome generated by long-read native RNA-sequencing, one of which is in the cytosolic receptor for dsDNA and known inflammatory inducible gene, Aim2. We show that this unannotated AFE isoform of Aim2 is the predominant isoform expressed during inflammation and contains an iron-responsive element in its 5'UTR enabling mRNA translation to be regulated by iron levels. This work highlights the importance of examining alternative isoform changes and translational regulation in the innate immune response and uncovers novel regulatory mechanisms of Aim2.

Not just anatomists but also clinicians from a variety of disciplines, including orthopedic doctors, surgeons, neurologists, and neurosurgeons, are interested in vertebral abnormalities. Numerous morphological anomalies, such as spinal synostosis, occipitalization, sacralization, and lumbarization, as well as the absence of the posterior vertebral arch, have been reported. Sequentially fused vertebral segments might result in spinal fusion, synostosis of the vertebrae, or blocked vertebrae. Movement limitations, early degenerative changes, and accompanying neurological abnormalities may all be brought on by the block vertebrae. The fusion may be whole or partial, acquired, or congenital. The cervical, lumbar, and thoracic vertebral levels are the most frequently affected by vertebral fusion, whether it is congenital or acquired. A systemic disorder, known as idiopathic diffuse idiopathic skeletal hyperostosis (DISH), is defined by distinctive ossification patterns that can run over at least three more vertebral levels or four successive vertebrae on their anterolateral aspect. Here, we present a series of spinal synostosis at various levels that were identified in museum specimens from our institution and that were associated with the clinical and developmental importance.

Abstract Background The lateral craniopharyngeal canal or Sternberg’s canal is located in the postero-lateral part of the sphenoid sinus. It is the weakest area of the skull base, originating due to defective fusion during embryological development in a small proportion of individuals. Hence, it can give rise to CSF leaks from that region. Due to the proximity of this region to the internal carotid artery, optic nerve, and cavernous sinus, it proves to be challenging to repair these defects. The exact location, role in the causation of CSF leaks, and clinical presentation with arachnoid herniation are also subject to much controversy. Results In our case series, we describe 7 patients with the rare clinical entity of CSF leak from the Sternberg’s canal, who had a mean age of 38.4 years, 4 of whom also had an arachnoid herniation through the canal. One patient already underwent prior endoscopic surgery for the CSF leak. We highlight a trans-nasal trans-sphenoid approach with multi-layer closure, i.e., fat plug, fascia, and a naso-septal flap with the use of an additional layer of muscle in one of the cases, following which all had good outcomes. Since the primary concern with this rare clinical scenario is correct identification and adequate exposure of the leak for repair, we additionally undertook a trans-pterygoid approach in 2 of our patients. Conclusions The key to the successful repair of the Sternberg’s canal leak lies in its accurate identification and exposure. In our series, we achieved good exposure by drilling the pterygoids and lateral sphenoid walls along with the use of angled endoscopes for visualization. Our case series highlights the linear relation of elevated BMI with spontaneous CSF leaks in the absence of benign intracranial hypertension and the reinforcement of repair with the use of a harvested naso-septal flap. Our multi-layered closure ensured good results and no recurrence of a leak in any of our patients.