EBioMedicine

Background: Alpha-1 antitrypsin deficiency (AATD) caused by the PI*ZZ mutation (Glu342Lys) results in hepatic accumulation of misfolded AAT-Z protein and reduced circulating AAT levels, leading to progressive liver disease and emphysema. Gene correction therapy represents a potentially curative approach by directly correcting the underlying genetic defect. We report the first case of successful hepatic gene correction with early histological and functional assessment. Methods/Case presentation: We report the case of a 66-year-old male patient with PI*ZZ AATD who underwent gene correction therapy within the YOLT-202 phase I/Ia clinical trial (clinical trial.gov ID NCT07193615). Ten weeks post treatment a liver biopsy was performed to re-evaluate pre-existing F2 liver fibrosis as measured by elastography before entering the study. Serum samples allowed functional assessment of the AAT-mediated elastase inhibition. Results: Liver biopsy did not show signs of hepatic inflammation and demonstrated 54% (Sanger) and 57% (Illumina) gene correction rate of the PI*ZZ variant on the DNA level with no bystander edits or off-target effects. Following a transient elevation of transaminases during the early post-treatment period, liver enzymes normalized. Monthly serum AAT measurements demonstrated biologically active and stable therapeutic levels throughout follow-up. Conclusions: This case demonstrates efficient and precise hepatic gene correction without concerning histological alterations and with substantial improvement of functional parameters, supporting the feasibility and safety of gene editing approaches for AATD.

Background Tuberculosis remains the leading infectious cause of death worldwide. In the WHO African region, declining incidence has coincided with antiretroviral therapy (ART) scale-up, though whether this reflects reduced progression to disease or reduced transmission is unclear. We evaluated how ART and symptom status influence within-household Mycobacterium tuberculosis complex (MTBC) transmission risk. Methods We conducted a case-contact household study in rural South Africa, enrolling index adults with bacteriologically-confirmed pulmonary tuberculosis. MTBC immunoreactivity was measured in all child household contacts (aged 2-14 years) as a proxy measure of within-household transmission. We assessed the influence of index person ART status and symptom status, and explored effect-measure modification of the association between index person HIV status and transmission risk by sex. Results Among 755 child contacts of 296 index persons, effective ART was not associated with within-household MTBC transmission risk (risk ratio [RR], 1.07; 95% CI, 0.66-1.74). Among PLHIV engaged in ART care, WHO TB four-symptom screen (WHO4SS) status was not associated with transmission risk (RR, 0.80; 95% CI, 0.43-1.47), although absence of reported cough reduced risk (RR, 0.61; 95% CI, 0.38-0.96). A pronounced interaction between sex and HIV status was observed: HIV-negative women had the highest within-household MTBC transmission risk (30.5% vs. 14.3% in women with HIV) whereas risks were similar between HIV-positive and HIV-negative men. Conclusions We found no evidence that effective ART or WHO4SS status influenced within-household MTBC transmission risk, though confidence intervals were wide. Absence of reported cough was associated with lower risk, and transmission risk was highest among child contacts of HIV-negative women. These findings suggest reported cough is a useful marker of transmission risk and that routine tuberculosis screening within ART care may reduce transmission from PLHIV; intensified efforts are nonetheless needed to achieve earlier tuberculosis detection in HIV-negative individuals.

Introduction: Synthetic oligopeptides provide a rapid and cost-efficient approach to developing antibodies and diagnostics for emerging viral variants. Methods: This study computationally and experimentally characterized a synthetic peptide analog of the SARS-CoV-2 spike subdomain 2 major disulfide loop (SD2MDL), designated S621 (CPVAIHADQLTPTWRVYSTC). Binding affinity was computationally estimated using the Heuristic Affinity Prediction Tool for Immune Complexes (HAPTIC), while experimental validation was performed using enzyme-linked immunosorbent assay (ELISA) with rabbit-derived antipeptide antibodies. Clinical diagnostic accuracy testing was done using plasma samples from RT-PCR-confirmed COVID-19 patients and pre-COVID-19 controls. Results: S621 demonstrated nanomolar binding affinity (Kdapp = 1.14 nM) and high avidity (3.67 nM), closely matching HAPTIC predictions (3.54 nM). Diagnostic evaluation yielded a sensitivity of 89.92% and specificity of 27.79%, corresponding to an overall accuracy of 71.79%. Discussion: These findings demonstrate that a single synthetic peptide derived from a conserved spike subdomain can function as a high-affinity surrogate for full-length antigens, supporting its potential application in rapid peptide-based immunodiagnostics.

Background: Neurological, neuropsychiatric, and neurodevelopmental disorders cluster in ALS families, sharing a common genetic architecture with ALS. Pathogenic variants in genes associated with other neurological, neurodevelopmental, or neuropsychiatric disorders may also co-occur in ALS and modify phenotype. We have sought to determine the prevalence and clinical pattern of likely-pathogenic/pathogenic (LP/P) non-ALS neurological, neurodevelopmental, and neuropsychiatric variants, alone and in combination with ALS-gene variants, in two large ALS cohorts. Methods: Whole-genome sequencing (WGS) of 469 Irish and 774 Answer ALS people with ALS (pwALS) was analysed for ClinVar LP/P variants associated with other neurological (n = 15541), neurodevelopmental (n = 9761), and neuropsychiatric (n = 321) phenotypes. Inheritance patterns for associated genes (autosomal recessive/autosomal dominant) along with the associated phenotype were validated using OMIM. Standardised clinical data included family history, site and age of onset, El Escorial category, survival, motor decline, and cognitive and behavioural assessments. Known ALS-gene variants and C9orf72 repeat expansion status were included for each cohort. Results: Non-ALS neurological variants were identified in 47/469 (10.0%) Irish and 69/774 (8.9%) Answer ALS participants, most frequently in hereditary spastic paraplegia-associated genes (3.2% Irish; 2.8% Answer ALS). Irish neurological variant carriers showed higher frequency of respiratory onset (10.6% vs 1.2%, Fisher's exact p = 0.002, {Phi} = 0.20) and fewer premorbid behavioural symptoms (0.92 +/- 0.56 vs 3.08 +/- 0.97, Cohen's d = -0.40). Neurodevelopmental variants occurred in 12/469 (2.6%) Irish and 20/774 (2.6%) Answer ALS participants. In the Irish cohort, neurodevelopmental variant carriers had significantly shorter survival in Cox proportional hazards model (log-rank p = 0.005), corresponding to a more than two-fold increased hazard of death (HR = 2.25, 95% CI 1.26-4.00), and had significantly increased familial burden of neuropsychiatric disorders among first- and second-degree relatives (negative binomial IRR for carriers = 2.41, 95% CI: 1.12-5.18, p = 0.025). Across combined cohorts, 18 individuals (Irish n = 8; Answer ALS n = 10) carried [≥]2 LP/P variants spanning ALS and non-ALS genes. Conclusion: Rare LP/P variants in genes associated with other neurological and neurodevelopmental disorders occur in up to 12% of pwALS across two independent cohorts. Carriers show distinct phenotypes, shorter survival, and characteristic family history patterns. These findings suggest that extended pleiotropic and oligogenic architectures may contribute to ALS heterogeneity.

Background World Symposium on Pulmonary Hypertension (WSPH) Group 2 pulmonary hypertension (PH) is a clinically integrated phenotype attributed to left heart disease, whereas pre- versus post-capillary classification is operationalized primarily by pulmonary capillary wedge pressure (PCWP). Although current recommendations emphasize contextual interpretation and provocative testing for intermediate PCWP values, the relationship between PCWP-based classification and underlying phenotype has not been systematically evaluated. We aim to quantify phenotype-hemodynamic discordance across the PCWP spectrum and evaluate a staged physiology-guided framework incorporating inhaled nitric oxide (iNO), ventricular geometry, and provocative testing. Methods We studied 1,032 participants from the NHLBI-sponsored PVDOMICS cohort with multidisciplinary adjudicated phenotypes integrating clinical, imaging, physiologic, and hemodynamic data. Stage-specific PCWP thresholds classified pre- versus post-capillary physiology at rest, during iNO, and during provocation (fluid challenge or invasive cardiopulmonary exercise testing [iCPET]). Echocardiographic right ventricular-to-left ventricular (RV/LV) ratio was evaluated as a marker of ventricular interdependence. Restricted cubic spline and staged concordance analyses defined certainty-based PCWP ranges and incremental diagnostic yield. Results Adjudicated Group 2 phenotype was present in 37.0% of participants. Resting PCWP demonstrated good discrimination (AUC 0.86), but substantial bidirectional phenotype-hemodynamic discordance persisted across intermediate PCWP ranges. At a resting PCWP of 12 mmHg, 25% of participants classified as pre-capillary had adjudicated Group 2 PH, whereas at 18 mmHg, 35% classified as post-capillary remained discordant non-Group 2. Concordance did not approach 90% until PCWP values were <9 mmHg or >24 mmHg. Dynamic testing incrementally improved concordance within these overlap zones. Nearly half of adjudicated Group 2 PH participants (46.5%) were not identified by resting PCWP alone; incorporation of iNO and provocative testing increased cumulative Group 2 identification by 63.4% and improved sensitivity from 79.9% to 83.7%. Model discrimination improved from an AUC of 0.863 to 0.908 (likelihood-ratio P<0.001). iNO increased PCWP in discordant Pre/G2 participants, unmasking latent left-sided limitation, while lowering PCWP in discordant Post/NonG2 participants, consistent with ventricular interdependence. RV/LV ratio [&ge;]0.94 reduced discordant Post/NonG2 classification by 70.5%, and incorporation of PCWP/cardiac output slope improved physiologic specificity during exercise. Conclusions Group 2 PH is a dynamic, load-dependent phenotype inadequately characterized by resting PCWP alone. Intermediate PCWP values represent continuous probabilities of bidirectional discordance rather than discrete diagnostic states. A staged physiology-guided approach integrating iNO, ventricular geometry, and provocative testing improves concordance between hemodynamic classification and clinically integrated phenotype assignment.

Amyotrophic lateral sclerosis (ALS) lacks a validated blood-based diagnostic, and the field is increasingly moving from single-molecule markers toward integrative, multi-component signatures. Here we present a liquid-biopsy strategy that transduces disease dependent serum-nanoparticle interactions into a learnable near-infrared spectral phenotype. A sensor array of twelve DNA-functionalized single-walled carbon nanotube (SWCNT) chiralities, functionalized with (GT)6 ssDNA coupled with a deep learning model was tested on serum from 20 ALS patients and 19 age- and sex-matched controls (n = 39, TargetALS). Our multiplexed sensor design (12 SWCNT chiralities) and data acquisition strategy based on excitation-emission matrices acquired at three timepoints (0, 6, 24 h) was conceived to maximize sensor carried information. Indeed, we show that the array generates partially independent temporal dynamics across chiralities governed primarily by tube diameter. To decode this multiplexed, time-resolved signal, we trained a dual-objective convolutional autoencoder that jointly optimizes reconstruction and classification, achieving 84.6% cross-validated accuracy (AUC = 0.87). Selected latent features were reproducible across an independent same-subject experimental batch and correlated with serum neurofilament light chain, linking the spectral phenotype to a clinically relevant neurodegeneration marker. Mass spectrometry supported a molecular basis for discrimination, revealing an ALS-biased protein corona enriched in adaptive-immune and inflammatory proteins. Together, these results establish proof of principle that time-resolved, multi-chirality SWCNT spectral sensing can compress complex serum composition into a reproducible near-infrared biomarker signature for ALS.

Background: Machine learning models trained on population health surveys offer scalable tools for cardiovascular screening, but recurring methodological weaknesses undermine their credibility and equity: data leakage from synthetic oversampling, qualitative rather than quantitative explainability evaluation, and the absence of demographic fairness auditing at the clinical operating threshold. Methods: We present EXHEART, a leakage-free stacked ensemble pipeline trained on BRFSS 2015 (n = 253,680) and validated on BRFSS 2020 (n = 319,795; temporal transport and retrain) and a clinical cardiovascular examination dataset (n = 68,730). The pipeline combines XGBoost, LightGBM, Random Forest, and a multi-layer perceptron as base learners with 5-fold out-of-fold logistic regression stacking and Platt scaling calibration. A quantitative SHAP-LIME consistency framework, based on Kendall-tau rank correlation and Jaccard overlap, accompanies a decision-curve analysis, a subgroup-stratified SHAP interaction analysis, and an intersectional fairness audit (Sex x Age x Income) with threshold-shifting mitigation and a frontier of the fairness-utility trade-off. The framework also adds cross-instrument fairness-disparity attribution, an empirical diagnostic that provides evidence on whether an observed subgroup disparity is more consistent with a measurement-induced or a substantive explanation by re-validating it on a dataset that measures the same clinical construct objectively. On heart disease, this diagnostic associates 89% of the sex TPR gap (95% CI [0.65, 0.99]) with the self-reported survey outcome rather than with a substantive risk difference. Results: On BRFSS 2015, EXHEART achieves AUC-ROC = 0.850, AUPRC = 0.371, Brier score = 0.071, and reduces ECE by 96% (0.256 to 0.011) via Platt scaling. Global SHAP-LIME rank agreement is moderate-to-strong (Kendall-tau = 0.580, Spearman-rho = 0.818) with a substantial top-3 divergence (Jaccard@3 = 0.200), where Stroke flips from SHAP rank 8 to LIME rank 1. The Sex TPR gap is 0.124 at the screening threshold; intersectional Sex x Age disparities reach 0.649 among adequately-powered cells, 5.2x the single-attribute gap. Temporal transport to BRFSS 2020 collapses sensitivity from 0.776 to 0.267, while retraining restores AUC = 0.840 and ECE = 0.012. On clinical examination data, the Sex TPR gap collapses to 0.014; the attribution test indicates this gap is instrument-dependent, consistent with a measurement or outcome-definition explanation rather than a substantive risk difference. Cross-domain SHAP analysis identifies four instrument-independent CVD risk factors and two major portability failures. Conclusions: EXHEART combines three practices that population-scale cardiovascular classifiers usually apply in isolation: leakage-free training with calibrated probabilities, a test of whether the model's explanations are stable, and a fairness audit that examines intersecting subgroups rather than single attributes. Bringing them together proved worthwhile. The intersectional audit revealed disparities that single-attribute auditing missed, and the cross-instrument comparison indicated that much of the sex gap reflects how the outcome is measured in survey data rather than a substantive difference in risk. The temporal transport findings indicate that deployed BRFSS models warrant periodic monitoring and retraining to maintain clinical utility. EXHEART is a retrospective methodological evaluation on public de-identified data; it is not validated for direct clinical decision-making, diagnosis, or treatment recommendation without prospective clinical validation.

Introduction: Variants in the polymerase gamma (POLG) gene are associated with a wide range of mitochondrial disorders. Emerging evidence suggests a potential link between POLG variants and Parkinson's disease (PD); yet, results remain inconclusive. Objectives: To investigate the genetic spectrum and prevalence of POLG variants in PD across diverse ancestries. Methods: We leveraged multi-ancestry genetic data from the Global Parkinson's Genetics Program (GP2), including genotyping data from 98,589 and short-read sequencing data from 36,022 individuals. We performed a POLG rare variant screen, case-control association, and gene-level burden analyses. Results: Five PD cases carried potentially biallelic rare pathogenic/likely pathogenic POLG variants. Additionally, 228 individuals (<1%; 161 PD cases, 28 individuals with other neurological disorders, and 39 controls) carried 34 distinct rare pathogenic/likely pathogenic heterozygous variants, with no significant frequency differences between cases and controls, except for the p.Ala467Thr variant in the European population. The co-inherited pathogenic variants p.Thr251Ile and p.Pro587Leu were present in <1% of both cases and controls, with no significant group differences. Burden and variant-level association analyses showed no association between rare POLG variant burden or common POLG variant enrichment and PD. Conclusions: POLG variants are overall rare in PD. The identification of rare pathogenic variants among PD cases suggests that POLG-related mitochondrial dysfunction may contribute to PD in isolated instances, particularly under recessive inheritance. Our findings support a role for POLG variants in select cases and underscore the need for larger-scale sequencing and functional studies.

Background: Cardiovascular-kidney-metabolic (CKM) syndrome is a leading driver of cardiovascular morbidity and mortality. Whole-body molecular imaging is well-positioned to phenotype such syndromes, yet no imaging biomarker quantifies cumulative CKM burden. Bone scintigraphy with 99mTc-labeled bisphosphonates is widely performed and expanding with transthyretin amyloidosis assessment, under which Perugini grade 0 (absent cardiac uptake) is considered clinically benign. Objective: We hypothesized that the soft tissue-to-bone ratio (STBR) on these scans captures CKM burden and is an independent prognostic biomarker. Methods: We retrospectively analyzed 8,769 consecutive patients without cardiac uptake on 99mTc-DPD whole-body planar scintigraphy. The primary endpoint was all-cause mortality. Secondary endpoints were major adverse cardiovascular events (MACE) and heart failure hospitalization. Cox models were adjusted for ten established cardiovascular risk factors. Imaging-phenotype association (IPA) analysis mapped STBR to 1,210 clinical traits. STBR distribution across CKM stages was assessed in four prespecified analyses, including a non-cancer subgroup. Results: During a median follow-up of 5.1 years (IQR 2.5-8.2), 2,418 deaths occurred. Patients with prespecified STBR >0.5 (n=772, 8.8%) had significantly higher mortality (adjHR 1.73, 95% CI 1.54-1.94, p<0.0001) with an adjHR of up to 3.42 at higher thresholds (95% CI 2.05-5.42, p<0.0001). Hazard increased monotonically with STBR. STBR >0.5 was independently associated with MACE (adjHR 1.51, 95% CI 1.11-2.05, p=0.008) and heart failure hospitalization (adjHR 1.31, 95% CI 1.02-1.67, p=0.03). The association was robust across all prespecified subgroups and sensitivity analyses, including continuous STBR and patients without renal insufficiency. IPA analysis identified significant associations with type 2 diabetes, chronic kidney disease, chronic ischaemic heart disease, heart failure, atrial fibrillation, liver disease, amyloidosis, and hypertension among binary traits, as well as with CRP, NT-proBNP, BUN, cholesterol (inverse), and hemoglobin (inverse) among continuous parameters. STBR increased monotonically across CKM stages in all sensitivity analyses (all p<0.0001). Conclusions: STBR derived from routine 99mTc-DPD bone scintigraphy in patients without cardiac uptake is an independent prognostic imaging biomarker associated with cumulative cardiovascular-kidney-metabolic burden. As an opportunistic measure from scans already acquired at scale, STBR could refine CKM risk stratification at no additional cost, radiation, or acquisition time.

Background. Capillary refill time, an examiner-dependent bedside test of distal microvascular perfusion, has become a resuscitation target in septic shock,1,2,3,4 motivating a continuous surrogate computed from the photoplethysmogram (PPG, the optical waveform the pulse oximeter on every ICU patient already records).5,6,7,8 Objective. We attempted three PPG-derived candidate measures on the MIMIC-IV Waveform Database (MIMIC-IV-WDB v0.1.0) and asked, by inspecting randomly drawn examples, whether each captured its intended physiology before any downstream modeling. Methods. MIMIC-IV-WDB v0.1.09 was linked to MIMIC-IV.10 The signals were a cuff-anchored perfusion-index recovery (reactive hyperemia when the cuff shares an arm with the probe), a slow Mayer-wave-band power ratio of the perfusion index (sympathetic vasomotor tone), and a per-beat diastolic exponential decay time constant (a refill-like recovery time). For each signal we drew 10 random examples at a fixed seed and checked them against a checklist fixed in advance. Each was read by the author and, separately, by MedGemma 1.5, a multimodal medical language model run locally. A synthetic test with a known time constant checked the third signal. Results. The cuff-anchored signal showed the expected occlusion-reperfusion shape on 268 of 6,236 evaluable cuff cycles (4.30%) in 15 of 19 patients, consistent with opposite-limb placement of the probe and cuff. The slow-band ratio returned a stable cohort value, but a clear, stationary peak appeared in only4 of 10 random windows. The per-beat fit met its goodness-of-fit threshold in 10 of 10 beats, yet a cardiac-frequency heuristic flagged a possible fit on the heart-rate oscillation in 7 of 10, and in 5 of 17 patients the time constant lay where an exponential is indistinguishable from a straight line. A 0.5Hz high-pass pre-filter implanted its own approximately 318 ms time constant regardless of truth. The language model tracked the human on clear positives but reported the pattern present on every call it returned, never absent. Conclusions. Two of the three candidate signals did not reflect their intended physiology in most examples, and the third was constrained by sensor placement. Inspecting a few random raw inputs against a checklist written in advance is an inexpensive upstream check before downstream inference on PPG-derived microvascular signals.

Synapse loss is an early feature of neurodegeneration and may provide sensitive biomarkers for experimental medicine. Positron emission tomography (PET) with the synaptic vesicle glycoprotein 2A radioligand [11C]UCB-J shows widespread signal reduction across dementias. However, it remains unclear which aspects of synaptic integrity [11C]UCB-J PET measures. We developed a histological-imaging pipeline to quantify structurally intact synapses in post-mortem brain tissue. We applied it to six donors with the tauopathy progressive supranuclear palsy (PSP) who had ante-mortem [11C]UCB-J-PET, alongside six controls across 11 brain regions. Synapse loss in PSP was widespread but region-specific across cortical, subcortical, and brainstem regions. Greater synapse loss was associated with higher tau burden and pathology, and cortical synaptic density correlated with ante-mortem cognition. Post-mortem synaptic density correlated with in vivo [11C]UCB-J-PET signal. This study provides validation of SV2A PET as a biomarker of synaptic density and supports integration of imaging with histopathology in neurodegenerative disease research.

Background. Fairness-aware machine learning increasingly targets demographic performance disparities in clinical prediction, yet whether standard bias mitigation strategies genuinely improve equity in physiological signal analysis remains unclear. Age-based disparities in photoplethysmography (PPG)-based heart rate prediction present a particular challenge, as age-related performance differences may reflect context-dependent physiological structure rather than correctable artifacts. Methods. We evaluated three fairness interventions, inverse-frequency weighting (IF), Group Distributionally Robust Optimization (GroupDRO), and adversarial debiasing (ADV), applied via fine-tuning of a PPG foundation model across three clinical datasets spanning intensive care unit, laboratory, and consumer wearable contexts. Outcomes were assessed using a 2x2 framework classifying each intervention-dataset combination by the joint direction of change in mean absolute error (MAE) and fairness gap (FG) across age groups, yielding four outcome types: genuine improvement (G), leveling down (L), selective benefit (S), and both worse (W). Results. Across nine intra-domain conditions, no intervention simultaneously improved both MAE and FG (0/9 genuine improvement). The dominant pattern was leveling down (5/9): FG decreased but was accompanied by MAE degradation, indicating that apparent fairness gains were achieved at the cost of overall predictive performance. Age-group difficulty ordering varied across clinical contexts at baseline and was not preserved under intervention. In 18 cross-domain transfer conditions, genuine improvement was rare (4/18) and observed exclusively in non-MIMIC source configurations; models fine-tuned on MIMIC-sourced data yielded no genuine improvements (0/6). Embedding-level representation changes following fine-tuning did not reliably predict fairness outcomes. Conclusions. Age-based fairness interventions in PPG heart rate prediction indicate a leveling-down pattern rather than genuine equity improvement, suggesting that age-related performance gaps reflect context-dependent physiological structure not fully addressable through standard bias mitigation. Cross-domain transfer further amplifies this instability. These findings suggest that fairness evaluation frameworks for age-stratified physiological prediction should account for context-dependent performance structure rather than treating observed gaps as correctable bias.

Background: Chronic low-grade inflammation drives cardiovascular-kidney-metabolic (CKM) syndrome. Clonal hematopoiesis of indeterminate potential (CHIP), an age-related driver of systemic inflammation, is linked to several cardiometabolic disorders. However, whether CHIP modifies CKM progression and contributes to heterogeneity in cardiovascular disease (CVD) risk within the CKM framework remains uninvestigated. Methods: This cohort study included 307,025 UK Biobank participants at CKM stages 0-3 free of baseline CVD. CHIP status was identified via whole-exome sequencing (WES). The association between CHIP and baseline CKM severity was examined, along with the independent and joint effects of CHIP and CKM stages on incident CVD risk. The joint effects of CHIP and polygenic risk scores (PRS) were further assessed, and the incremental predictive value of incorporating CHIP into the AHA PREVENT equations was evaluated. Results: CHIP carriers were more likely to present with advanced CKM stages [OR 1.14 (1.09-1.20), P < 0.001] and exhibited higher incident CVD risk during follow-up [HR 1.13 (1.08-1.18), P < 0.001]. Significant joint effects between CHIP and CKM stages were observed, with the highest risk among CHIP carriers at CKM stage 3 [HR 1.63 (1.50-1.78), P < 0.001]. Large or multiple CHIP mutations conferred greater hazards, with distinct gene-specific effects observed. Moreover, CHIP and high genetic risk also jointly amplified CVD susceptibility. Most importantly, incorporating CHIP into AHA PREVENT significantly improved risk discrimination. Conclusions: CHIP is a significant risk factor associated with more advanced CKM stages and amplifies incident CVD risk. Integrating CHIP into existing prevention strategies may refine CVD risk stratification.

Brain-derived tau (BD-tau) is an emerging blood-based biomarker for neurodegeneration, yet there are currently limited well validated BD-tau assays available for research and clinical use. To enhance access to this vital biomarker for neurological disorders including traumatic brain injury (TBI), we developed a novel blood-based immunoassay for BD-tau on the ultra-sensitive Quanterix HD-X platform using Single Molecule Array technology. Analytical validation assessed dilution linearity, specificity, precision, detection limits, and spike recovery, each recording robust metrics in agreement with international expert recommendations. The assay demonstrated robust validation metrics, achieving between-run stability of 95% when analyzing aliquots from six independent plasma and serum samples across five analytical runs. It also showed strong dilution linearity when diluted four-fold and achieved over 90% recovery when spiked with cerebrospinal fluid. Next, we evaluated the clinical utility of the assay in cohorts of individuals with traumatic brain injury (TBI), where strong performances were recorded whether using the 2-step or 3-step assay formats ({rho}= 0.94; p < 0.0001). Furthermore, plasma BD-tau distinguished samples from TBI patients based on time from injury and severity (AUC=0.93). Plasma BD-tau differentiated between favorable and unfavorable functional outcomes in the acute-severe group. Our findings underscore the significant potential of the BD-tau assay as a biomarker for TBI in the severe phase.

Vaccines to prevent infant group B streptococcus (GBS) disease are advancing, with licensure likely based on safety and immunologic endpoints rather than clinical efficacy data. This approach requires robust, generalisable serological thresholds of risk reduction (SToRRs). We combined data from six case-control studies in Europe and Africa to define SToRRs for early-onset (EOD) and late-onset (LOD) GBS disease. Across diverse epidemiological and healthcare settings, anti-capsular polysaccharide IgG concentrations were consistently higher in infants who remained disease free than in those who developed disease. Higher antibody concentrations were required to reduce the risk of EOD than LOD, and higher concentrations were required for serotype Ia than for serotype III. This study provides a quantitative framework to support correlates-based evaluation and potential licensure of maternal GBS vaccines.

As genetic testing becomes increasingly integrated into Parkinson disease (PD) research, including targeted testing for variants in LRRK2 and GBA1, the return of individual research results is becoming more common. However, limited qualitative data exists regarding how research participants experience genetic results disclosure and post-test genetic counseling in PD research settings. We conducted semi-structured qualitative interviews with participants (n=13) enrolled in the Parkinson Precision Medicine Initiative (formerly Parkinson Progression Markers Initiative; PPMI) who had received PD-related genetic test results and post-test genetic counseling. Interviews were conducted 1 to 3 weeks following result disclosure and analyzed using thematic analysis with a primarily deductive coding approach informed by study aims and inductive identification of emergent themes. Four primary themes were identified: (1) personal connection and motivations for participation, (2) centrality of result disclosure and information preferences, (3) emotional experiences and support needs, and (4) communication quality and alignment with participant needs. Overall, our findings underscore the importance of person-centered genetic counseling within PD research. As return of genetic and biomarker results in research and clinical trial contexts expand, thoughtful integration of relational, informational, and communication-focused practices will be essential to support participant engagement and trust.

Background Coeliac disease (CD) diagnosis on duodenal biopsies is limited by interobserver variability. We have previously demonstrated pathologist-level performance with our artificial intelligence (AI) model for the histopathological diagnosis of adult CD, but not in paediatric practice. As paediatric CD screening programmes expand internationally, accurate and scalable diagnostic tools are needed. We investigated whether an AI model trained exclusively on adult whole-slide images (WSIs) can generalise to paediatric CD diagnosis across independent centres. Methods A training and validation dataset of 9,958 WSIs from 8,421 adult patients (961 CD) from five centres was used to develop an ensemble of multiple-instance learning models using features from a foundation model. Testing was performed on 708 consecutive paediatric patients (86 CD) from two centres (Edinburgh and Southampton) not included in training. Model calibration was assessed, and probability outputs were grouped into clinically interpretable categories. Findings In adult cross-validation, the AI model achieved an area under the receiver operating characteristic curve (AUC) of 98.7%, sensitivity of 84.9%, specificity of 99.0%, and negative predictive value (NPV) of 98.1%. On testing (paediatric) datasets, performance remained high (AUC 98.8%, sensitivity 80.2%, specificity 98.4%, NPV 97.3%). Restricting analysis to predictions outside the intermediate-probability range (predicted CD probability <10% or [&ge;]65%; 85.3% of cases) improved sensitivity to 100% and specificity to 98.7%. No misclassifications were observed among high-confidence predictions (<2% or [&ge;]85%; 66.0% of cases). The expected calibration error was 0.03. Performance improved significantly when biopsies from both duodenal sites (bulb [D1] and descending [D2/3]) were considered. Interpretation Our AI model, trained on adult biopsies, generalises to paediatric CD diagnosis across centres and scanner platforms. Well-calibrated probability outputs provide clinically interpretable measures of diagnostic confidence and could support safe identification of CD-negative biopsies within defined thresholds. These findings demonstrate the feasibility of applying adult-derived AI models in paediatric populations and reinforce the importance of multi-site (D1 & D2) biopsy sampling.

Background: Mono-allelic Dehydrodolichyl Diphosphate Synthase (DHDDS) variants are associated with juvenile Parkinsonism, developmental delay and seizures. Symptoms are progressive, and various mechanisms, such as defective glycosylation, lysosomal dysfunction and cholesterol accumulation have been hypothesized to underlie disease symptoms. There is no treatment for DHDDS-related disease. Methods: Patient-derived cortical forebrain organoids were created to elucidate disease mechanisms and evaluate potential treatments. In these neuronal models, glycosylation, lipidomics, proteomics, cholesterol/ganglioside accumulation, mitochondrial function and electrophysiological activity were assessed. Finally, we investigated the effects of nicotinamide mononucleotide (NMN), identified through a yeast-based drug screen, in neuronal cell models and in six patients in an off-label, N-of-1, observational series. Results: DHDDS-patient derived organoids showed visual signs of degeneration after four months of culturing. This was accompanied by significant cholesterol accumulation in astrocytes, decreased mitochondrial respiration and loss of deep-layer neurons. In addition, we identified glycosylation abnormalities, showing for the first time that glycosylation in human tissue is affected by monoallelic DHDDS variants. Proteomic analysis revealed altered protein expression of proteins involved in lipid metabolism, cytoskeletal organization and neuronal development. We found that oral Nicotinamide Mononucleotide supplementation led to significant improvement in mitochondrial respiration and electrophysiological parameters in organoids, concurring with clinical improvements in all of the treated patients, particularly regarding their ataxia and tremor. Conclusion: Our findings reveal a progressive phenotype in DHDDS-patient-derived brain organoids, with mitochondrial dysfunction and astrocyte-specific metabolic alterations contributing to disease pathology. Notably, NMN treatment led to clinical improvements in patients with heterozygous DHDDS variants, highlighting its potential as a therapeutic strategy.

Background and Aims: MASLD has become the most prevalent chronic liver disease globally. Although MVPA and plasma fatty acids have been individually studied in relation to metabolic health, their independent and combined associations with MASLD incidence remain unclear. We aimed to investigate these associations. Methods: This study included 51,717 UK Biobank participants free of liver disease at baseline, with MVPA measured using wrist-worn accelerometers and plasma fatty acids quantified via NMR. Multivariable-adjusted Cox models and restricted cubic splines were used. Results: Over a median follow-up of 7.8 years, 472 incident cases were identified. In fully adjusted models, meeting recommended MVPA levels together with higher n-6 PUFA concentrations was associated with a 71% lower risk (HR 0.29, 95% CI 0.18-0.45). The MVPA-MASLD association was nonlinear, with risk reduction plateauing at approximately 189 minutes per week. Higher n-6 PUFA was associated with reduced risk, whereas n-3 PUFA showed no significant association. Conclusions: These findings suggest that behavioral and metabolic factors may jointly influence MASLD risk. Further studies in diverse populations are needed to confirm these associations.

Background: Treatment decisions for carotid atherosclerotic disease rely primarily on luminal stenosis, although plaque vulnerability and symptomatic status better reflect short-term cerebrovascular risk. A scalable CTA tool for automated phenotyping of symptomatic carotid disease is lacking. Materials & Methods: In this multi-institutional retrospective study, 689 patients (mean age, 67.9 {+/-} 7.7 years; 366 men) from four hospitals were analyzed after screening 705 CTA examinations. 423 patients from one center were used for five-fold development and internal validation, and 266 patients from three centers for independent external validation. CarotidMamba, a deep learning framework combining dual foundation-model encoders with Mamba-based sequence modeling, was developed and benchmarked against clinical, radiomics, clinic-radiomics, CNN, and transformer comparators. Results: In the development cohort, CarotidMamba achieved an AUC of 0.839 (95% CI, 0.799-0.879) and accuracy of 0.825 (95% CI, 0.793-0.857), outperforming the strongest comparator by 0.066 and 0.050, respectively. External validation yielded AUCs of 0.897 (95% CI, 0.835-0.959) in YCH, 0.809 (95% CI, 0.720-0.898) in DCH, and 0.762 (95% CI, 0.649-0.875) in GH-NTC. CarotidMamba showed the lowest Brier score and expected calibration error across cohorts, with calibration slopes near 1.0. Conclusion: CarotidMamba provides an interpretable, clinically oriented, and externally validated CTA framework for phenotyping symptomatic carotid plaques, supporting vulnerability-aware imaging assessment beyond stenosis alone.