Peptide ligands for the universal purification of exosomes by affinity chromatography.

Exosomes are gaining prominence as vectors for drug delivery, vaccination, and regenerative medicine. Owing to their surface biochemistry, which reflects the parent cell membrane, these nanoscale biologics feature low immunogenicity, tunable tissue tropism, and the ability to carry a variety of payloads across biological barriers. The heterogeneity of exosomes' size and composition, however, makes their purification challenging. Traditional techniques, like ultracentrifugation and filtration, afford low product yield and purity, and jeopardizes particle integrity. Affinity chromatography represents an excellent avenue for exosome purification. Yet, current affinity media rely on antibody ligands whose selectivity grants high product purity, but mandates the customization of adsorbents for exosomes with different surface biochemistry while their binding strength imposes elution conditions that may harm product's activity. Addressing these issues, this study introduces the first peptide affinity ligands for the universal purification of exosomes from recombinant feedstocks. The peptides were designed to (1) possess promiscuous biorecognition of exosome markers, without binding process-related contaminants and (2) elute the product under conditions that safeguard product stability. Selected ligands SNGFKKHI and TAHFKKKH demonstrated the ability to capture of exosomes secreted by 14 cell sources and purified exosomes derived from HEK293, PC3, MM1, U87, and COLO1 cells with yields of up to 80% and up-to 50-fold reduction of host cell proteins (HCPs) upon eluting with pH gradient from 7.4 to 10.5, recommended for exosome stability. SNGFKKHI-Toyopearl resin was finally employed in a two-step purification process to isolate exosomes from HEK293 cell fluids, affording a yield of 68% and reducing the titer of HCPs to 68 ng/mL. The biomolecular and morphological features of the isolated exosomes were confirmed by analytical chromatography, Western blot analysis, transmission electron microscopy, nanoparticle tracking analysis.

AI-based automated segmentation for ovarian/adnexal masses and their internal components on ultrasound imaging.

Purpose: Segmentation of ovarian/adnexal masses from surrounding tissue on ultrasound images is a challenging task. The separation of masses into different components may also be important for radiomic feature extraction. Our study aimed to develop an artificial intelligence-based automatic segmentation method for transvaginal ultrasound images that (1) outlines the exterior boundary of adnexal masses and (2) separates internal components. Approach: A retrospective ultrasound imaging database of adnexal masses was reviewed for exclusion criteria at the patient, mass, and image levels, with one image per mass. The resulting 54 adnexal masses (36 benign/18 malignant) from 53 patients were separated by patient into training (26 benign/12 malignant) and independent test (10 benign/6 malignant) sets. U-net segmentation performance on test images compared to expert detailed outlines was measured using the Dice similarity coefficient (DSC) and the ratio of the Hausdorff distance to the effective diameter of the outline ( R HD - D ) for each mass. Subsequently, in discovery mode, a two-level fuzzy c-means (FCM) unsupervised clustering approach was used to separate the pixels within masses belonging to hypoechoic or hyperechoic components. Results: The DSC (median [95% confidence interval]) was 0.91 [0.78, 0.96], and R HD - D was 0.04 [0.01, 0.12], indicating strong agreement with expert outlines. Clinical review of the internal separation of masses into echogenic components demonstrated a strong association with mass characteristics. Conclusion: A combined U-net and FCM algorithm for automatic segmentation of adnexal masses and their internal components achieved excellent results compared with expert outlines and review, supporting future radiomic feature-based classification of the masses by components.

Non-Breast Implantable Medical Devices and Associated Malignancies: A Systematic Review.

Innovation in healthcare has led to the development of numerous implantable medical devices (IMDs). However, advances in our knowledge of breast implant-associated malignancies have raised questions about the prevalence, etiology and management of malignancies associated with non-breast IMDs. The objective of this study was to examine the prevalence and characteristics of malignancies associated with non-breast IMDs. An expert medical librarian developed the search strategy for this review. Databases included MEDLINE (National Library of Medicine, Bethesda, MD; date range of search: 1946 to June 21, 2023), EMBASE (Elsevier, Amsterdam, the Netherlands; date range of search: 1946 to June 21, 2023), and Cochrane Central Register of Controlled Trials (CENTRAL; Cochrane Library, Wiley, Hoboken, NJ; date range of search: 1946 to June 21, 2023). In addition, grey literature sources were searched and relevant references from systematic reviews and meta-analyses were included. The PRISMA guideline was used to guide the review. Risk of bias was evaluated using the JBI Critical Appraisal tools. A total of 12,230 articles were reviewed with a total of 77 meeting inclusion criteria. Risk of bias was highest with case reports (moderate, average of 65.1% with range of 37.5 to 100%) and low for the remaining study types. In total, 616 cases of IMD-associated malignancies were identified. Malignancies associated with IMDs were reported in the head/neck (543, 88.1%), lower extremity (57, 9.6%), thorax (9, 1.4%), abdomen (3, 0.5%), and genitourinary system (2, 0.3%). The most common malignancy type in the lower extremity was sarcoma, in the head and neck was squamous cell carcinoma, and in the thorax was lymphoma. This study is the first comprehensive systematic review of its kind. Overall, the oncologic risk of IMDs is low. The discussion of malignancy is an important part to the overall consent process and malignancy should be considered with any new signs or symptoms in the anatomic area of an implant. More data is needed to better understand how primary malignancies occur around IMDs and how to reduce this risk.

A hippocampal circuit mechanism to balance memory reactivation during sleep.

Memory consolidation involves the synchronous reactivation of hippocampal cells active during recent experience in sleep sharp-wave ripples (SWRs). How this increase in firing rates and synchrony after learning is counterbalanced to preserve network stability is not understood. We discovered a network event generated by an intrahippocampal circuit formed by a subset of CA2 pyramidal cells to cholecystokinin-expressing (CCK+) basket cells, which fire a barrage of action potentials ("BARR") during non-rapid eye movement sleep. CA1 neurons and assemblies that increased their activity during learning were reactivated during SWRs but inhibited during BARRs. The initial increase in reactivation during SWRs returned to baseline through sleep. This trend was abolished by silencing CCK+ basket cells during BARRs, resulting in higher synchrony of CA1 assemblies and impaired memory consolidation.

Eligibility for Substance Use Clinical Trials Among Emergency Psychiatry Patients: The Impact of Exclusion Criteria.

Objective: The first objective was to identify common exclusion criteria used in clinical trials. The second objective was to quantify the degree to which these criteria exclude emergency psychiatry patients. Methods: Qualitative Content Analysis was used for the first objective, identifying common exclusion criteria used in recent high-impact substance use clinical trials. A retrospective record review was used for the second objective, which examined the frequency of these exclusion criteria in a 1-month sample of adults receiving psychiatric evaluation in an emergency department. Results: Most trials had exclusions for co-occurring psychiatric problems (76.6%), medical problems (74.0%), prior or current treatment (72.7%), motivation for change (61.1%), pregnancy or lactation (57.1%), or using other specified substances of abuse (54.6%). In the clinical sample, exclusions for co-occurring psychiatric problems would make 94.7% of patients ineligible. Other exclusions had a combined effect of making 76% of patients ineligible. Conclusions: Clinical trials using typical exclusion criteria exclude nearly all emergency psychiatry patients with substance use problems.

Fully automated determination of robotic pedicle screw accuracy and precision utilizing computer vision algorithms.

Historically, pedicle screw accuracy measurements have relied on CT and expert visual assessment of the position of pedicle screws relative to preoperative plans. Proper pedicle screw placement is necessary to avoid complications, cost and morbidity of revision procedures. The aim of this study was to determine accuracy and precision of pedicle screw insertion via a novel computer vision algorithm using preoperative and postoperative computed tomography (CT) scans. Three cadaveric specimens were utilized. Screw placement planning on preoperative CT was performed according to standard clinical practice. Two experienced surgeons performed bilateral T2-L4 instrumentation using robotic-assisted navigation. Postoperative CT scans of the instrumented levels were obtained. Automated segmentation and computer vision techniques were employed to align each preoperative vertebra with its postoperative counterpart and then compare screw positions along all three axes. Registration accuracy was assessed by preoperatively embedding spherical markers (tantalum beads) to measure discrepancies in landmark alignment. Eighty-eight pedicle screws were placed in 3 cadavers' spines. Automated registrations between pre- and postoperative CT achieved sub-voxel accuracy. For the screw tip and tail, the mean three-dimensional errors were 1.67 mm and 1.78 mm, respectively. Mean angular deviation of screw axes from plan was 1.58°. For screw mid-pedicular accuracy, mean absolute error in the medial-lateral and superior-inferior directions were 0.75 mm and 0.60 mm, respectively. This study introduces automated algorithms for determining accuracy and precision of planned pedicle screws. Our accuracy outcomes are comparable or superior to recent robotic-assisted in vivo and cadaver studies. This computerized workflow establishes a standardized protocol for assessing pedicle screw placement accuracy and precision and provides detailed 3D translational and angular accuracy and precision for baseline comparison.

High-Throughput Covalent Modifier Screening with Acoustic Ejection Mass Spectrometry.

Interests in covalent drugs have grown in modern drug discovery as they could tackle challenging targets traditionally considered "undruggable". The identification of covalent binders to target proteins typically involves directly measuring protein covalent modifications using high-resolution mass spectrometry. With a continually expanding library of compounds, conventional mass spectrometry platforms such as LC-MS and SPE-MS have become limiting factors for high-throughput screening. Here, we introduce a prototype high-resolution acoustic ejection mass spectrometry (AEMS) system for the rapid screening of a covalent modifier library comprising ∼10,000 compounds against a 50 kDa-sized target protein─Werner syndrome helicase. The screening samples were arranged in a 1536-well format. The sample buffer containing high-concentration salts was directly analyzed without any cleanup steps, minimizing sample preparation efforts and ensuring protein stability. The entire AEMS analysis process could be completed within a mere 17 h. An automated data analysis tool facilitated batch processing of the sample data and quantitation of the formation of various covalent protein-ligand adducts. The screening results displayed a high degree of fidelity, with a Z' factor of 0.8 and a hit rate of 2.3%. The identified hits underwent orthogonal testing in a biochemical activity assay, revealing that 75% were functional antagonists of the target protein. Notably, a comparative analysis with LC-MS showcased the AEMS platform's low risk of false positives or false negatives. This innovative platform has enabled robust high-throughput covalent modifier screening, featuring a 10-fold increase in library size and a 10- to 100-fold increase in throughput when compared with similar reports in the existing literature.

A wound-induced differentiation trajectory for neurons.

Animals capable of whole-body regeneration can replace any missing cell type and regenerate fully functional new organs, including new brains, de novo. The regeneration of a new brain requires the formation of diverse neural cell types and their assembly into an organized structure with correctly wired circuits. Recent work in various regenerative animals has revealed transcriptional programs required for the differentiation of distinct neural subpopulations, however, how these transcriptional programs are initiated in response to injury remains unknown. Here, we focused on the highly regenerative acoel worm, Hofstenia miamia, to study wound-induced transcriptional regulatory events that lead to the production of neurons and subsequently a functional brain. Footprinting analysis using chromatin accessibility data on a chromosome-scale genome assembly revealed that binding sites for the Nuclear Factor Y (NFY) transcription factor complex were significantly bound during regeneration, showing a dynamic increase in binding within one hour upon amputation specifically in tail fragments, which will regenerate a new brain. Strikingly, NFY targets were highly enriched for genes with neuronal function. Single-cell transcriptome analysis combined with functional studies identified soxC+ stem cells as a putative progenitor population for multiple neural subtypes. Further, we found that wound-induced soxC expression is likely under direct transcriptional control by NFY, uncovering a mechanism for the initiation of a neural differentiation pathway by early wound-induced binding of a transcriptional regulator.

Synthesis of Zintl Phase Metal Silicide Thermoelectric Materials in Magnesium/Zinc Flux.

Zintl phases have potential applications as thermoelectric materials for power generation and cooling owing to their complex crystal structures and unique electronic properties. We carried out reactions of silicon with barium and strontium in excess Mg/Zn flux to synthesize (Ba/Sr)5+xMg19-xSi12 Zintl phases, investigating the effect of varying Ba/Sr ratio on site mixing and thermoelectric properties. (Ba/Sr)5+xMg19-xSi12 compounds with 0 < x < 3 are charge-balanced Zintl phases which adopt the hexagonal Ho5Ni19P12 structure type (space group P6̅2m). Density of states calculations indicate that these materials are semimetals. Single-crystal X-ray diffraction data and elemental analysis for Ba5Mg19Si12, Ba4.86Sr2.94Mg16.20Si12, Ba3.63Sr4.20Mg16.17Si12, Ba1.93Sr5.99Mg16.08Si12, and Sr7.82Mg16.18Si12 show occupation of barium and strontium cations in Ho sites, while strontium mixes with magnesium on a specific Ni site. Powder XRD data of products show that they are single phase throughout the sample. Thermoelectric measurements indicate that increasing strontium content and mixing on three cation sites decreases thermal conductivity; it is hypothesized that improved overall thermoelectric behavior is likely due to the rattling of the Sr cations in their positions.

The crosstalk between fibroblast growth factor 21 (FGF21) system and substance use.

Existing literature indicates that communication between the central nervous system and the peripheral nervous system is disrupted by substance use disorders (SUDs), including alcohol use disorder (AUD). Fibroblast growth factor 21 (FGF21), a liver-brain axis hormone governing energy homeostasis, has been shown to modulate alcohol intake/preference and other substances. To further elucidate the relationship between FGF21, alcohol use, and other substance use, we conducted a scoping review to explore the association between FGF21 and SUDs. Increases in FGF21 reduce alcohol consumption while suppressing FGF21 increases alcohol consumption, demonstrating an inverse relationship. Alcohol elevates FGF21 levels primarily via the liver, subsequently promoting neuronal signals to curb alcohol intake. FGF21 activation engages molecular pathways that defend against alcohol-induced fat accumulation, oxidative stress, and inflammation. Considering the bidirectional association between FGF21 and alcohol, further studies on the FGF21 system as a potential pharmacotherapy for AUD and alcohol-associated liver disease are warranted.

Increasing physical activity among adults affected by COVID-19 social distancing restrictions: A feasibility trial of an online intervention.

In response to COVID-19 social distancing restrictions, digitally delivered health interventions present as a potential solution for maintaining or improving individuals' physical activity. This study explored the feasibility of a web-based intervention, informed by the multi-process action control (M-PAC) framework to promote PA among individuals affected by social distancing. Fifty adults self-reporting as insufficiently active were randomized to a 6-week web-intervention (n = 27) or wait-list control (n = 23). Primary feasibility outcomes included recruitment and retention rates and usability and satisfaction scores; secondary outcomes of MVPA and M-PAC constructs and tertiary outcomes of mental health and wellbeing were also assessed. Overall, feasibility of the intervention was high, with a 96% recruitment rate, 84% retention rate, high satisfaction and usability scores, and comparable website usage to similar eHealth interventions. Intervention participants trended towards improved MVPA and M-PAC constructs and outcomes of mental health and wellbeing. Findings suggest study extension to a full-scale RCT.

Morphology of the scaphotrapeziotrapezoid joint: A multi-domain statistical shape modeling approach.

The scaphotrapeziotrapezoid (STT) joint is involved in load transmission between the wrist and thumb. A quantitative description of baseline STT joint morphometrics is needed to capture the variation of normal anatomy as well as to guide staging of osteoarthritis. Statistical shape modeling (SSM) techniques quantify variations in three-dimensional shapes and relative positions. The objectives of this study are to describe the morphology of the STT joint using a multi-domain SSM. We asked: (1) What are the dominant modes of variation that impact bone and articulation morphology at the STT joint, and (2) what are the morphometrics of SSM-generated STT joints? Thirty adult participants were recruited to a computed tomography study of normal wrist imaging and biomechanics. Segmentations of the carpus were converted to three-dimensional triangular surface meshes. A multi-domain, particle-based entropy system SSM was used to quantify variation in carpal bone shape and position as well as articulation morphology. Articular surface areas and interosseous proximity distributions were calculated between mesh vertex pairs on adjacent bones within distance (2.0 mm) and surface-normal angular (35°) thresholds. In the SSM, the first five modes of variation captured 76.2% of shape variation and contributed to factors such as bone scale, articular geometries, and carpal tilt. Median interosseous proximities-a proxy for joint space-were 1.39 mm (scaphotrapezium), 1.42 mm (scaphotrapezoid), and 0.61 mm (trapeziotrapezoid). This study quantifies morphological and articular variations at the STT joint, presenting a range of normative anatomy. The range of estimated interosseous proximities may guide interpretation of imaging-derived STT joint space.

Influenza Vaccine Effectiveness Against Influenza A-Associated Emergency Department, Urgent Care, and Hospitalization Encounters Among US Adults, 2022-2023.

BACKGROUND: The 2022-2023 United States influenza season had unusually early influenza activity with high hospitalization rates. Vaccine-matched A(H3N2) viruses predominated, with lower levels of A(H1N1)pdm09 activity also observed. METHODS: Using the test-negative design, we evaluated influenza vaccine effectiveness (VE) during the 2022-2023 season against influenza A-associated emergency department/urgent care (ED/UC) visits and hospitalizations from October 2022 to March 2023 among adults (aged ≥18 years) with acute respiratory illness (ARI). VE was estimated by comparing odds of seasonal influenza vaccination among case-patients (influenza A test positive by molecular assay) and controls (influenza test negative), applying inverse-propensity-to-be-vaccinated weights. RESULTS: The analysis included 85 389 ED/UC ARI encounters (17.0% influenza A positive; 37.8% vaccinated overall) and 19 751 hospitalizations (9.5% influenza A positive; 52.8% vaccinated overall). VE against influenza A-associated ED/UC encounters was 44% (95% confidence interval [CI], 40%-47%) overall and 45% and 41% among adults aged 18-64 and ≥65 years, respectively. VE against influenza A-associated hospitalizations was 35% (95% CI, 27%-43%) overall and 23% and 41% among adults aged 18-64 and ≥65 years, respectively. CONCLUSIONS: VE was moderate during the 2022-2023 influenza season, a season characterized with increased burden of influenza and co-circulation with other respiratory viruses. Vaccination is likely to substantially reduce morbidity, mortality, and strain on healthcare resources.

A web-based physical activity intervention targeting affect regulation: a randomized feasibility trial.

Early career professionals (ECPs) are a critical target for physical activity (PA) promotion. Affect contributes to an established PA intention-behaviour gap and is pertinent among ECPs. OBJECTIVE: The purpose of this study was to examine the feasibility and acceptability of a web-based intervention and explore the effects on secondary outcomes (moderate-to-vigorous PA (MVPA), emotion regulation, multi-process action control constructs). METHODS: Adults aged 25-44 who were employed at least part-time in a desk-based job and not meeting PA guidelines (<150 min MVPA) were recruited and randomized into a 6-week online intervention integrating acceptance and commitment principles and affect regulation strategies, or a control group. RESULTS: Forty adults were recruited and randomized to the web-based intervention (n = 21) and waitlist control (n = 19). The recruitment rate was 29%, retention was 75%, engagement was 68%, and satisfaction was high in both quantitative and qualitative assessment. Participants allocated to the intervention improved MVPA (ηp2=0.30), emotion regulation (ηp2 =0.49), behavioural regulation (ηp2=0.53), affective attitude (ηp2=0.23), identity (ηp2=0.24), and constructs of mindfulness (ηp2=0.44), and valued living (ηp2=0.20). CONCLUSIONS: Primary outcomes concerning feasibility were adequate and secondary outcomes improved, suggesting a full-scale randomized controlled trial is feasible with minor modifications. A large-scale study is warranted to establish intervention effectiveness.

National trends and ecological factors of physical activity engagement among U.S youth before and during the COVID-19 pandemic: A cohort study from 2019 to 2021.

BACKGROUND: This study aims to investigate the trends and ecological determinants of physical activity among U.S. children and adolescents during the 2019-2021 period, encompassing the COVID-19 pandemic's onset and subsequent years. METHODS: Utilizing data from the National Survey of Children's Health over three years, this cohort study analyzed physical activity levels and ecological determinants among 82,068 participants aged 6-17. The sample included 36,133 children (44%) and 45,935 adolescents (56%), with variables assessed by caregiver reports. RESULTS: The analysis revealed a significant decline in physical activity among children from 2019 to 2020, followed by a recovery in 2021, whereas adolescents showed a continued decrease without recovery. Over the study period, children were consistently more active than adolescents. Better health status, normal weight, less screen time, stronger peer relationships, higher parental involvement, better family resilience and greater school participation were consistently correlated with increased physical activity in both age groups. Sleep duration was a predictor of activity only in children, while mental health status was solely a predictor in adolescents. Neighborhood environment consistently predicted children's activity levels but was a significant factor for adolescents only in 2020. CONCLUSIONS: These findings highlight the differing impacts of the pandemic on physical activity between children and adolescents, emphasizing the need for targeted public health interventions, particularly for adolescents whose activity levels have not recovered from the pandemic period. Age-specific physical activity interventions should consider sleep duration and neighborhood environmental factors when targeting children and mental health factors when focused on adolescents.

Structural switch in acetylcholine receptors in developing muscle.

During development, motor neurons originating in the brainstem and spinal cord form elaborate synapses with skeletal muscle fibres1. These neurons release acetylcholine (ACh), which binds to nicotinic ACh receptors (AChRs) on the muscle, initiating contraction. Two types of AChR are present in developing muscle cells, and their differential expression serves as a hallmark of neuromuscular synapse maturation2-4. The structural principles underlying the switch from fetal to adult muscle receptors are unknown. Here, we present high-resolution structures of both fetal and adult muscle nicotinic AChRs, isolated from bovine skeletal muscle in developmental transition. These structures, obtained in the absence and presence of ACh, provide a structural context for understanding how fetal versus adult receptor isoforms are tuned for synapse development versus the all-or-none signalling required for high-fidelity skeletal muscle contraction. We find that ACh affinity differences are driven by binding site access, channel conductance is tuned by widespread surface electrostatics and open duration changes result from intrasubunit interactions and structural flexibility. The structures further reveal pathogenic mechanisms underlying congenital myasthenic syndromes.

Consistency and variability in multimodal parent-child social interaction: An at-home study using head-mounted eye trackers.

Real-time attention coordination in parent-toddler dyads is often studied in tightly controlled laboratory settings. These studies have demonstrated the importance of joint attention in scaffolding the development of attention and the types of dyadic behaviors that support early language learning. Little is known about how often these behaviors occur in toddlers' everyday lives. We brought wireless head-mounted eye trackers to families' homes to study the moment-to-moment patterns of toddlers' and parents' visual attention and manual activity in daily routines. Our sample consisted of English- and Spanish-speaking families who all reported being middle- or upper middle-class. Toddlers were 2 to 3 years old. Consistent with the findings from previous laboratory studies, we found variability in how frequently toddlers attended to named objects in two everyday activities-Object Play and Mealtime. We then tested whether parent-toddler joint attention in the seconds before a naming utterance increased toddler's attention on the named object. We found that joint attention accompanied by the attended object being held increased the child's attention to the labeled object during naming. We posit that in the rich, noisy world of toddlers' everyday lives, embodied attention plays a critical role in coordinating dyadic behaviors and creating informative naming moments. Our findings highlight the importance of studying toddlers' natural behavior in the real world. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Clinical Course of TGA After Arterial Switch Operation in the Current Era.

Background: The number of patients with an arterial switch operation (ASO) for transposition of the great arteries (TGA) is steadily growing; limited information is available regarding the clinical course in the current era. Objectives: The purpose was to describe clinical outcome late after ASO in a national cohort, including survival, rates of (re-)interventions, and clinical events. Methods: A total of 1,061 TGA-ASO patients (median age 10.7 years [IQR: 2.0-18.2 years]) from a nationwide prospective registry with a median follow-up of 8.0 years (IQR: 5.4-8.8 years) were included. Using an analysis with age as the primary time scale, cumulative incidence of survival, (re)interventions, and clinical events were determined. Results: At the age of 35 years, late survival was 93% (95% CI: 88%-98%). The cumulative re-intervention rate at the right ventricular outflow tract and pulmonary branches was 36% (95% CI: 31%-41%). Other cumulative re-intervention rates at 35 years were on the left ventricular outflow tract (neo-aortic root and valve) 16% (95% CI: 10%-22%), aortic arch 9% (95% CI: 5%-13%), and coronary arteries 3% (95% CI: 1%-6%). Furthermore, 11% (95% CI: 6%-16%) of the patients required electrophysiological interventions. Clinical events, including heart failure, endocarditis, and myocardial infarction occurred in 8% (95% CI: 5%-11%). Independent risk factors for any (re-)intervention were TGA morphological subtype (Taussig-Bing double outlet right ventricle [HR: 4.9, 95% CI: 2.9-8.1]) and previous pulmonary artery banding (HR: 1.6, 95% CI: 1.0-2.2). Conclusions: TGA-ASO patients have an excellent survival. However, their clinical course is characterized by an ongoing need for (re-)interventions, especially on the right ventricular outflow tract and the left ventricular outflow tract indicating a strict lifelong surveillance, also in adulthood.