Disease Models & Mechanisms

High throughput sequencing necessitates robust computational tools to contextualize complex transcriptomic data. In this study, we analyzed RNA Seq data from peripheral blood of bacterial pneumonia patients and healthy controls (GEO GSE161731) to investigate systemic molecular responses to infection. Differential gene expression analysis identified significantly upregulated genes in pneumonia. We used GANGO for Gene Ontology (GO) term enrichment and BioFunctional for hierarchical exploration, providing an initial overview of affected biological processes. To deepen interpretation, we leveraged CURIE (www.curieai.tech ), an AI plat-form that cross-references GO terms with scientific literature. CURIE generates narrative explanations, link-ing molecular mechanisms, experimental evidence, and publications to each biological process, moving be-yond simple enrichment. Our combined analysis revealed a coordinated upregulation of pathways involved in DNA repair and replication, transcriptional activation, RNA processing, translation, ribosome biogenesis, mitochondrial respiration, and cofactor binding. By integrating these findings within a broad, literature-supported framework, we highlighted how multiple cellular compartments and processes collaborate to mount an effective response to infection-induced stress. This study underscores the potential of combining classical statistical enrichment with AI based interpretation for a holistic understanding of host-pathogen in-teractions at the transcriptomic level.

Background: Cervical spinal cord (CSC) compression and myelopathy are frequent, severe complications in mucopolysaccharidosis (MPS) types I and VI. Early detection is critical to prevent irreversible neurological deficits and preserve quality of life. This prospective study aimed to identify quantitative neuroimaging biomarkers of structural CSC damage and myelopathy in MPS I and VI. Methods: Structural and diffusion magnetic resonance imaging (MRI) of the CSC was analyzed in 17 MPS I patients, 3 MPS VI patients (19{+/-}7 years), and 19 case-matched healthy controls (20{+/-}7 years). CSC macrostructure (cross-sectional area, diameters) and microstructure (fractional anisotropy [FA], radial [RD], mean, axial diffusivities) were evaluated within and beyond the maximal compression areas. Results: MPS patients demonstrated reduced CSC cross-sectional area and anterior-posterior diameter at upper cervical levels, alongside widespread microstructural alterations (lower FA, higher RD) throughout the CSC versus controls (pFDR<0.05). Restricted foramen magnum space was associated with microstructural damage at lower cervical levels without overt macrostructural compression. Morphological and microstructural metrics were sensitive to CSC compression and myelopathy severity (pFDR<0.05). Lower CSC integrity correlated with more advanced growth retardation (p<0.001). Among patients receiving hematopoietic stem cell transplantation, earlier treatment correlated with preserved upper CSC morphology (p<0.05). Distinct neuroimaging phenotypes were observed across MPS genotypes (pFDR<0.05). Conclusions: CSC MRI provides candidate neuroimaging biomarkers for early detection and characterization of myelopathy in MPS I and VI. Structural damage extends beyond primary maximal compression areas, highlighting these markers for clinical monitoring and future therapeutic trials.

Background Cardiac amyloidosis is an increasingly recognized cause of heart failure in older adults, yet Colombian data, and data from the southwest of the country in particular, remain scarce. We describe the clinical, diagnostic, imaging and outcome profile of patients with cardiac amyloidosis at two high-complexity institutions in Cali. Methods Observational, descriptive, retrospective, two-center study. Patients were identified in the institutional registries of both centers using ICD-10 codes for amyloidosis, and clinical data were abstracted manually from medical records. Continuous variables are reported as median (interquartile range) and categorical variables as counts and percentages over the number of patients with available data. Results Thirty-two patients were analyzed (median age 77 years; 68.8% men). Heart failure (90.6%), atrial fibrillation (46.9%) and carpal tunnel syndrome (31.2%) were frequent. The etiological classification depended on the test considered: bone scintigraphy was positive (Perugini 2-3) in 23 of 26 patients studied (88.5%), but a monoclonal protein was screened for in only 22, and the full non-invasive criteria for ATTR were met in 16 of 32 (50.0%). Genetic testing was positive in 9 of 18 tested, and the variant was Val142Ile in all 8 patients in whom it was characterized. The working diagnosis was ATTR in 26 patients (81.2%) and AL in 6 (18.8%). Tafamidis was used in 37.5%. Eleven of the 31 patients with known vital status died (35.5%), and NT-proBNP was markedly higher among those who died (6640 vs 1186 pg/mL; p=0.003). Conclusions In this southwestern Colombian cohort, cardiac amyloidosis presented as the classic transthyretin phenotype, with the Val142Ile variant in all 8 patients in whom it was characterized. Bone scintigraphy was frequently positive, but screening for a monoclonal protein was often omitted, so only half the cohort met the full non-invasive criteria for ATTR. Mortality was high and closely associated with NT-proBNP.

Abstract Many women experience menopausal symptoms after stopping hormone replacement therapy (HRT). An evidence gap remains regarding how women and healthcare professionals (HCPs) make decisions about discontinuing and restarting HRT. Aim To explore experiences of HCPs that support women when they discontinue and restart HRT. Design and setting Qualitative study with HCPs working in NHS primary care. Method Semi-structured interviews with HCPs (n = 15) conducted between October 2025 and March 2026. A preliminary coding template was created using the discussion guide as a framework. Each interview was then coded inductively against the template which developed iteratively. Team based discussion helped refine codes into themes and sub-themes, and a subsequent conceptual model was created. Results The model describes HCPs experiences of supporting women to discontinue HRT as an iterative process characterised by uncertainty. Discussions about discontinuation were commonly prompted by changes in clinical risk or annual HRT review, but decisions about whether and when to stop also depended on womens symptoms, side effects, life stage and readiness. HCPs described agreeing an individualised discontinuation strategy, providing support during the process, and reviewing women after discontinuation to consider alternative treatments or, where appropriate, restarting HRT. Conclusion HCPs recognise the importance of readiness to discontinue and that decision making should be woman led, with HCP support, clear practice guidance and opportunities to revisit treatment. Further research is needed to support women weigh up their personalised risk and benefit of continuing/discontinuing and identify discontinuation strategies that limit resurgent symptoms.

Autistic adults face considerable challenges in receiving adequate support, from both paid/formal (such as from disability services) and unpaid/informal (for example, from friends and family) sources. Having adequate support can lead to better health and well-being outcomes. However, most instruments designed to assess adequacy of support were not made specifically for/with autistic adults. This paper presents a psychometric validation of the AASPIRE Adequacy of Supports and Information Scale (AASIS), a community-informed adaptation of a previously developed instrument (the Quality of Care and Support scale; Lucas-Carrasco et al., 2011), designed to be relevant and accessible to autistic adults. We also used the AASIS to explore correlates of perceptions of support in a sample of 798 autistic adults. Our confirmatory factor analysis supported a two factor structure for the AASIS (factor one: Supports; factor two: Information). Further testing confirmed the instrument was invariant across several groups, including race/ethnicity, gender, intellectual disability, communication fluency, and self versus proxy-report. Regression analyses suggested that the way in which participants joined the study (i.e., through community advertisements or via established healthcare/disability services) impacted whether they viewed their support as adequate. Additionally, a higher number of adverse childhood events and greater experience of discrimination were related to worse perceptions of support adequacy across both factors. Our findings support the preliminary validity of the AASIS across a variety of autistic people, though further testing is warranted. Findings also point to the importance of addressing the needs of autistic people who are minoritized and/or who have complex trauma histories.

Background Autism spectrum disorder (ASD) shows wide phenotypic variation, from adults without intellectual disability and with low support needs to minimally verbal children with intellectual disability and severe behavioral symptoms. Genetic studies of this heterogeneity have focused mainly on high-impact de novo variants and polygenic scores; here we investigate, at the molecular level, the role of common variants. Materials and methods We analyzed genotypic and phenotypic data from 9,332 individuals with ASD in the SPARK cohort. We considered 31 traits: six latent factors derived from the Social Communication Questionnaire and the Repetitive Behavior Scale-Revised, 22 co-occurring diagnoses, parent-reported IQ range, age at diagnosis, and age at first walking. We performed within-case genome-wide association studies (GWAS) and, for the 10 traits with evidence of SNP heritability, association analyses with genetically predicted gene and protein expression, splicing, and enhancer activity in brain and blood (xWAS). Nominated genes were tested for enrichment in biological processes, brain cell types, and developmental stages. xWAS-derived transcriptional signatures were compared with chemical perturbation signatures from CMap/LINCS L1000 to prioritize candidate drugs. Results GWAS identified seven genome-wide significant loci for five traits, including loci near GABRG3 and TENM2 associated with co-occurring sleep, feeding or toileting problems. xWAS identified seven gene-trait associations involving three phenotypes (FDR < 0.1), including CACNA1G with communication skills and NEGR1 with learning disability. GWAS and xWAS nominated different genes which however converged on synaptic transmission and gated channel activity, with preferential expression in neurons during later stages of brain development. PARP and p38 MAPK inhibitors were predicted to reverse the signatures of both social-communication and repetitive-behavior phenotypes, whereas several compound classes, including histone deacetylase inhibitors and mTOR inhibitors, were predicted to reverse the signatures of some phenotypes while mimicking those of others. Conclusion Combining within-case GWAS with xWAS nominates candidate genes mediating common-variant effects on ASD heterogeneity, provides biological insight, and suggests candidates for phenotypically stratified drug repositioning.

Background: Hereditary spastic paraplegia (HSP) comprises a heterogeneous group of neurodegenerative disorders characterized by progressive lower-extremity weakness and spasticity, with or without additional neurological symptoms. Objectives: To characterize the molecular genetic spectrum, inheritance patterns, molecular finding rate, and clinical characteristics of HSP within a multicenter Canadian cohort. Methods: We investigated 671 patients from 423 families who met clinical criteria for HSP identified through the Canadian HSP (CanHSP) registry from 7 medical centers across Canada. Patients were evaluated using known HSP and HSP mimic gene panels, whereas unsolved and more recent patients underwent whole-exome sequencing (WES). 435 individuals underwent gene-panel testing and 485 underwent WES. Copy-number variants (CNVs) were assessed using ExomeDepth, and candidate CNVs using multiplex ligation-dependent probe amplification (MLPA). Results: Molecular findings were identified in 426 of 671 individuals (63.5%): 351 (52.3%) had HSP-associated findings, 70 (10.4%) HSP-mimicking disorders, and five (0.7%) other molecular findings; 245 (36.5%) remained unresolved. Candidate variants were found in 80 relevant genes, 36 of which were HSP-associated genes. The most frequently implicated genes were SPAST (22.5%), SPG7 (5.7%), ATL1 (5.2%), SPG11 (3.6%), and SPG8 (3.3%). Phenotypically, SPAST, ATL1, and SPG8 w[zg1.1]ere predominantly associated with pure HSP, whereas SPG7 and SPG11 more often showed complicated phenotypes, with prominent cerebellar involvement in SPG7 and cognitive and developmental features in SPG11. Conclusion: Patients in CanHSP present with genetic and phenotypic heterogeneity. Reanalysis and complementary approaches, including whole-genome, RNA, and long-read sequencing, may increase diagnostic yield.

Introduction: Thyroid diseases are major health problems and among the most common endocrine disorders worldwide, including in Ethiopia. They are characterized by alterations in hormone secretion, enlargement of the thyroid gland, or both. Fine needle aspiration cytology is an important initial screening tool, and histopathologic examination is essential for the definitive diagnosis of thyroid disease. This study aimed to compare the diagnostic accuracy of fine needle aspiration cytology with histopathology in identifying thyroid diseases. Methods: A cross-sectional study was conducted at three health facilities in Hawassa City, Ethiopia, involving 304 patients who underwent both thyroid fine needle aspiration cytology and histopathologic examination of the same site between 30th October 2024 and 29th October 2025. FNAC results were classified using the Bethesda system, with histopathology as the gold standard. Data were entered into Kobo Toolbox, exported to SPSS version 27, and analyzed using cross-tabulation and chi-square ({chi}{superscript 2}) tests to determine diagnostic accuracy. Result: 304 patient records were included in the study. The fine needle aspiration cytology was determinate/diagnostic in 81.6% of cases. The study revealed the sensitivity and specificity of fine needle aspiration cytology to be 88.2% (CI 77.6-95.1%) and 94.4% (CI 90.6-97.1%), respectively. The false negative and false positive rates in this study were 2.4% and 4.4%, respectively. The positive predictive value was 80.3% (CI 68.7-89.3%), and the negative predictive value was 96.8% (CI 93.8-98.7%). The accuracy of fine needle aspiration cytology in differentiating benign from malignant thyroid lesions was 93.1%. Conclusion: FNAC demonstrated strong agreement with histopathology and remains a reliable, cost-effective first-line diagnostic tool for thyroid disease, particularly in resource-limited settings. Strengthening sample handling, ultrasound guidance, and adherence to Bethesda criteria may further improve diagnostic precision

Mitochondrial disease arises from variants in either the mitochondrial or nuclear genome that impair mitochondrial oxidative phosphorylation. Complex III (ubiquinol:cytochrome c oxidoreductase, CIII2) is a key enzyme of the mitochondrial respiratory chain, catalyzing electron transfer from reduced coenzyme Q10 (ubiquinol) to complex IV, via cytochrome c. CIII2 has one subunit encoded by mitochondrial DNA, the remainder being nuclear encoded. Individuals with CIII2 deficiency present with significant clinical heterogeneity, on account of pathogenic variants in genes encoding structural CIII2 subunits or assembly factors. UQCC1 (GenBank: NM_018244.5) encodes the mitochondrial complex III assembly factor Ubiquinol-cytochrome c reductase complex assembly factor 1, which is currently not associated with human disease. Here we describe nine subjects from eight unrelated families harboring biallelic UQCC1 variants with heterogeneous, early-onset mitochondrial disease presentations and lactic acidosis. Subject-derived fibroblasts demonstrated a decrease in the steady-state levels of UQCC1 and its interacting partner UQCC2, with structural modelling supportive of variant pathogenicity. BN-PAGE and quantitative proteomics demonstrated decreased CIII2 abundance whilst mass spectrometry (MS)-complexome profiling corroborated these findings, identifying impaired respiratory supercomplex formation and a delay in the incorporation of newly synthesized CIII2 subunits into native assemblies. Lentiviral transduction experiments in subject-derived fibroblasts restored the levels of both UQCC1 protein and CIII2 activity. Collectively, these findings provide convergent genetic, biochemical, proteomic and functional evidence that bi-allelic UQCC1 variants disrupt CIII2 assembly, establishing UQCC1 deficiency as a cause of mitochondrial CIII2 deficiency.

Genetic regulation of neurodevelopmental disorders that impair brain development and social interaction is complex and poorly understood. Although emerging evidence suggests a link between neurodevelopmental disorders and autophagy deficits, congenital autophagy gene disorders are rare in humans, and underlying molecular mechanisms remain unknown. Here we demonstrate that the autophagy protein BECN1 is a regulator of neuronal and social development in vertebrate species. We identify rare deleterious biallelic BECN1 variants that cause neurodevelopmental disorders in two unrelated families. Becn1 disruption also causes neuronal and social developmental defects in mouse and zebrafish models. BECN1 serves as an adaptor to recruit Arc to the E3 ubiquitin ligase UBE3A for polyubiquitination and degradation, thereby maintaining postsynaptic AMPA receptor (AMPAR) stability, AMPAR-mediated excitatory neurotransmission, and normal social interaction. Thus, our study illustrates new biology and a non-canonical mechanism by which BECN1 regulates neurodevelopment in humans, mice and zebrafish, via a crosstalk with the ubiquitin-proteasomal machinery.

X-linked adrenoleukodystrophy (X-ALD) is the most common monogenic disorder affecting brain white matter, presenting a broad clinical spectrum from slowly progressive adrenomyeloneuropathy (AMN) to rapidly advancing cerebral demyelination. Factors influencing the variable disease expressivity remain poorly understood. We conducted shotgun metagenomic profiling of fecal samples from X-ALD patients and integrated these data with targeted plasma metabolomic analyses of amino acids and key metabolic intermediates. We identified distinct microbiome-associated metabolic signatures across X-ALD phenotypes. Notably, Bacteroides cellulosilyticus and Helicobacter apodemus abundance were significantly altered in AMN compared with childhood CALD patients. In parallel, X-ALD patients exhibited evidence of microbial metabolic remodeling characterized by altered amino acid metabolism, reduced folate biosynthetic potential, and enrichment of stress-adaptive pathways, including redox-associated functions, glyoxylate shunt activity, lipid A modification, lipopolysaccharide assembly, biofilm adhesin biosynthesis, and curli fiber production. Consistent with these microbial changes, plasma levels of branched-chain amino acids and methionine, as well as serum folate concentrations, were significantly reduced. Importantly, methionine levels showed an inverse correlation with neurofilament light chain (NfL) concentrations in patients with cerebral disease. Our findings reveal a previously unrecognized link between gut microbial metabolic capacity and systemic metabolic alterations in X-ALD. Our results reinforce previous evidence implicating methionine metabolism in disease severity. Collectively, these data support a modulatory role of the microbiota-gut-brain axis in neuroinflammatory and mitochondrial-related processes underlying X-ALD progression and highlight new opportunities for biomarker development and microbiome-informed therapeutic interventions.

Hypothyroidism has been associated with increased skin cancer risk, whereas greater genetic susceptibility to hypothyroidism is associated with lower risk. These opposing associations suggest that the average risk associated with hypothyroidism may conceal substantial heterogeneity among patients, and that genetic background could help identify those at highest risk. In this study, we analyzed more than 650,000 individuals across All of Us and FinnGen cohorts to determine whether a hypothyroidism polygenic risk score (H-PRS) stratifies skin cancer risk among patients with autoimmune hypothyroidism and whether integration of a skin cancer polygenic risk score (SC-PRS) further refines risk. In FinnGen, autoimmune hypothyroidism was associated with increased skin cancer hazard (hazard ratio [HR] = 1.10; 95% CI [1.06, 1.15]), whereas H-PRS was inversely associated with risk (HR=0.97 per 1-SD increase; 95% CI [0.95, 0.98]). The hypothyroidism x H-PRS interaction was also negative (HR=0.94; 95% CI [0.91, 0.98]), indicating greater hypothyroidism-associated risk at lower H-PRS. The hypothyroidism and H-PRS associations replicated in All of Us, and similar H-PRS risk gradients were observed for melanoma, basal cell carcinoma, and squamous cell carcinoma in both cohorts. Among hypothyroid patients, predicted 10-year risk increased with higher SC-PRS and lower H-PRS. Combined clinical-genetic profiles had HRs ranging from 2.32 at the 20th H-PRS/80th SC-PRS percentiles to 3.37 at the 5th/95th percentiles, exceeding published point estimates for the established dermatological risk factors. Among women aged 55, representing the more commonly affected sex at approximately the median age of diagnosis, 10-year risk of skin cancer was 14.9% (95% CI [8.4%, 20.9%]) in the highest-SC-PRS/lowest-H-PRS quartiles versus 2.8% (95% CI [1.0%, 4.7%]) in the opposite quartiles. Overall, these findings support PRS-guided risk stratification for targeted skin cancer prevention and surveillance.

Artemisinin partial resistance (ART R) associated Plasmodium falciparum Kelch13 mutations is expanding across Africa. However, their population level consequences remain poorly understood, particularly in settings of moderate to high malaria transmission. We investigated associations between Kelch13 and partner drug mutation prevalence and monthly confirmed malaria incidence across 16 Ugandan districts from 2017 to 2022, using integrated molecular, epidemiological, climatic, malaria control, and antimalarial resistance data and negative binomial regression. Combined prevalence of WHO validated and candidate Kelch13 mutations increased over the study period. Each 10 percentage point increase in Kelch13 prevalence was associated with a 13.1% higher incidence after multivariable adjustment (aIRR 1.131, 95% CI 1.090 to 1.174). Furthermore, each 10% increase in pfcrt K76T was associated with a 24% increase in malaria transmission. As expected, ITN distribution >80%, IRS coverage >90%, and higher altitude were associated with lower incidence, whereas temperatures of 25 to 30C, relative humidity of 60 to 80%, and higher precipitation were associated with higher incidence. The persistence of the Kelch13 and pfcrt K76T association after adjustment suggests that expanding ART R associated parasite populations are independently associated with malaria incidence. The relationship may be bidirectional. These findings support interpreting ART R within its ecological and intervention context and integrating molecular, epidemiological, and malaria control surveillance to safeguard antimalarial efficacy.

Sickle cell disease (SCD) results from homozygosity or compound heterozygosity for a single point mutation in the beta hemoglobin gene. Despite carrying the same underlying causal mutation, patients exhibit highly heterogeneous clinical outcomes. Acute chest syndrome (ACS) is one of the leading causes of mortality in SCD. In this study, we aimed to identify genetic variants associated with ACS occurrence and recurrence (>5 episodes) in two SCD cohorts (OMG-SCD and Walk-PHaSST) of predominantly African ancestry and characterize their downstream molecular consequences. Although none of the SNPs reached the genome-wide threshold in the occurrence meta-analysis, we identified 40 independent genetic loci with p <10-5. However, the recurrence GWAS in OMG-SCD, which compared individuals with no prior history of ACS to individuals with a history of >5 episodes, identified one genome-wide significant locus along with 38 other independent loci with p <10-5. Survival analysis suggested lower survival probability among young ACS recurrence cases compared to controls or occurrence cases. Furthermore, molecular QTL analysis of ACS recurrence identified 42 SNP-protein and eight SNP-metabolite associations with p <10-3. Pathway and enrichment analysis identified secretory, platelet/hemostatic, and complement-mediated pathways, confirming the complex and heterogeneous pathophysiology of ACS. This study highlights the importance of a well-defined phenotype for identification of risk loci and elucidates the complex interplay of hemolytic, inflammatory, and cellular organization pathways that warrants further evaluation through large-scale multi-omic studies of SCD patients. Ultimately, the more extreme phenotype of ACS recurrence may be a more informative phenotype for analysis, risk prediction, and therapeutic targeting.

Several polyglutamine-encoding repeat expansions cause spinocerebellar ataxias (SCAs) and other movement disorders. SCAs are genetically heterogeneous with a substantial diagnostic gap. We used genome sequencing data from 266 individuals of European and Asian ancestry and screened genome-wide for repeat expansions using STRling, a dedicated repeat expansion caller. The cohort comprised patients with movement disorders, predominantly dystonia and ataxia, as well as unaffected participants. Further, we screened another 436 individuals with ataxia and 427 ethnically matched controls through fragment analysis of the most promising candidate locus in EP400. We applied targeted long-read sequencing to expansion carriers and one control individual to resolve the sequence structure of the repeat locus. Finally, we used microsatellite markers in chromosome 12q24.32-33 to test for a common founder. A genome-wide outlier analysis in STRling revealed a polyglutamine-encoding repeat expansion (GRCh38 chr12:132,062,525-132,062,590) in the E1A binding protein p400 (EP400) gene in two patients with ataxia. Through screening of additional ataxia patients, we identified in total 18 patients from 14 families with heterozygous repeat expansions in EP400. All expansion carriers were from Serbia. The repeat number in affected individuals ranged from 37 to 55, whereas the longest control allele had 32 repeats. We demonstrated that non-expanded alleles carried a CAA interruption within the 5-prime CAG tract; this interruption was lost in expanded alleles. Expansion carriers have pure SCA with onset mainly in their 40s and 50s. There is an inverse correlation between the number of repeats and age at onset. The EP400-encoded protein plays an essential role in chromatin remodeling, epigenetic regulation and DNA repair. Pathogenic sequence variants in EP400 have previously been implicated in a neurodevelopmental disorder. Our study identifies repeat expansions in EP400 as a novel and frequent cause of ataxia in Serbian patients. This finding further increases the diagnostic yield of ataxia testing and warrants investigation in ataxia patients worldwide. We add EP400 to the growing list of ataxia-associated repeat loci, which warrants inclusion in diagnostic ataxia panels.

Objective: Amphetamine (AMP) and its derivates are commonly prescribed for the management of attention-deficit/hyperactivity disorder (ADHD). This study aimed to 1) quantify AMP's transfer into human milk in lactating women receiving therapeutic doses and 2) contextualize infant exposure. Methods: Milk samples and dyad health histories of thirteen lactating women treated with varying doses of either mixed AMP, dextroamphetamine, or lisdexamfetamine were released for this study from the InfantRisk Human Milk Biorepository. Milk drug concentrations were quantified by validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) and used to calculate standard relative infant dose (RIDmat) and RID standardized to the defined daily dose (RIDDDD) of 15 mg AMP base. Results: Average milk concentrations (Cavg) of AMP base ranged from 23.3 to 297.0 ng/mL and increased linearly (R^2 = 0.92) with maternal daily dose. The RIDmat ranged 3.6-10.8% and RIDDDD ranged 1.4-21.8%. The fitted regression line crossed the 10% RIDDDD threshold at 1.17x DDD, with the upper 95% prediction limit crossing at 0.84x DDD. No adverse event reported by the mother-infant dyads were attributed to AMP exposure, notably including absence of reduced milk supply. Conclusions:The levels of AMP observed in human milk followed a dose predictable manner. The average patient was predicted to remain below the 10% RID DDD safety threshold at total daily doses at or below approximately 28 mg of mixed AMP salts or 59 mg of lisdexamfetamine. Ninety-five percent of patients were predicted to remain below the threshold at or below approximately 20 mg and 40 mg, respectively.

Background: Plasma biomarkers offer a minimally invasive alternative for detecting Alzheimer disease (AD) pathology, yet the genetic determinants influencing their levels remain largely underexplored. Objective: To identify genetic variants associated with AD-related plasma biomarker levels in a Midwestern Amish cohort. Methods: We analyzed plasma levels of amyloid beta (A{beta}42/A{beta}40), phosphorylated tau 181 (p-tau181), and the p-tau181/A{beta}42 ratio in the Midwestern Amish. Genome-wide association studies (GWAS) were used to examine associations with common genetic variants, while genetic linkage analysis was used to assess the potential impact of rare variants. Results: GWAS found associations between plasma biomarkers and the APOE gene. Additionally, novel candidate genes were implicated, including MME for A{beta}42/A{beta}40 ratio, and VEGFC and LRRTM4 for p-tau181. Linkage analysis identified significant signals for p-tau181 on chromosome 1, potentially involving the CSF1 and AHCYL1genes. Conclusions: The novel genetic variants associated with AD pathology implicate metabolic, synaptic, neuronal, and immune pathways. The use of GWAS and linkage analysis sheds light on the genetic architecture of AD-related plasma biomarkers.

Background: The relationship between recent viral infection and subsequent susceptibility to, or protection against, other viruses remains unclear, particularly across ages and symptom status. Methods: A prospective cohort study enrolled participants aged 6 months to 49 years in Washington and Oregon, USA, 2022-2024. Weekly nasal swabs were tested for SARS-CoV-2, respiratory syncytial virus (RSV), and influenza A/B; additional swabs were collected during symptomatic illness and were also tested for parainfluenza (PIV) 1-4, endemic human coronaviruses (hCOV), adenovirus, human metapneumovirus, human rhinovirus (hRV), and enteroviruses. Proportional hazards regression models were used to characterize associations between incident symptomatic infection (illness) and history of recent illness due to other viruses (within 1-7, 8-14, 15-30, 31-60, 61-90, 91-180 days), adjusted for geographic region, age, sex, daycare/school/work attendance, household size, and number of children in the household. For SARS-CoV-2, influenza, and RSV, these analyses were repeated to study incidence of infection (including asymptomatic cases). Findings: Of 3,560 participants, 2,969 (83%) had [&ge;]1 illness over 4,299 person-years. Adjusted hazard ratios (aHRs) in the short term ([&le;]2 weeks) after an illness were predominantly below 1, indicating that recent viral illness was often associated with lower risk of subsequent illness with other viruses. Across age groups, recent hRV illness was associated with lower risk of hCOV, PIV, and RSV illness (aHRs 0.07-0.27). SARS-CoV-2 illness was associated with a higher short-term risk of adenovirus and PIV illness (aHRs 3.20-3.33). In children aged 6 months-4 years, recent influenza illness was associated with higher short-term risk of SARS-CoV-2 illness. Including asymptomatic infections generally attenuated these associations. Interpretation: Some viral illnesses may alter short-term susceptibility to subsequent viral illnesses. Interactions may differ by age and symptom status. Funding: US Centers for Disease Control and Prevention; Washington State Department of Health.

BACKGROUND: The chromosome 9p21.3 locus is among the strongest genetic risk loci for cardiovascular disease (CVD), yet the regulatory mechanisms underlying this association remain poorly understood. Previous vascular disease studies focused predominantly on the role of this locus in smooth muscle cell (SMC) dysfunction, whereas its potential role in endothelial cells (ECs) remains less well characterized. Here, we sought to identify candidate regulatory variants at 9p21.3, their distal target genes, and effector transcripts that contribute to CVD-related phenotypes in ECs. METHODS: Promoter capture Hi-C was used to map chromatin interactions at 9p21.3 in primary human ascending aortic SMCs (n=6), supplementing the previously obtained ECs (n=16) dataset. To support the regulatory relevance of these interactions, expression quantitative trait locus analysis was performed in 136 vascular biopsies. Variant effects were prioritized in silico using AlphaGenome. The regulatory activity of the prioritized region was validated using CRISPR activation. RESULTS: Chromatin interaction mapping identified a complex regulatory architecture at the 9p21.3 locus involving 19 putative regulatory elements and 15 genes across ECs and SMCs. MIR31HG showed the most recurrent distal promoter connectivity with the 9p21.3 region. Consistent with the chromatin interaction map, expression of MIR31HG transcripts was associated with genetic variation at the 9p21.3 locus. In ECs, the lncRNA MIR31HG, but not miR-31, was responsive to hemodynamic flow. Depletion of MIR31HG in ECs was associated with endothelial activation. Epigenetic tracks and deep learning predictions highlighted an active enhancer element at 9p21.3, which was subsequently validated by CRISPR activation and increased MIR31HG expression, confirming its capacity to regulate MIR31HG over a large genomic distance of approximately 538 kb. CONCLUSIONS: Altogether, these data implicate lncRNA MIR31HG as a likely effector of the 9p21.3 CVD locus, linking the cis-regulatory element and heritable genetic variation in the respective region to endothelial activation through long-range enhancer regulation.

Background: Artificial intelligence (AI) may improve electrocardiographic differentiation of Takotsubo syndrome from acute anterior ST-segment elevation myocardial infarction (STEMI), but its impact on human diagnostic performance, the added value of visual explanations, and differences by reader background and experience remain uncertain. Methods: Using 12-lead electrocardiogram (ECG) images from two hospitals, we developed and externally validated an ensemble of three deep learning models after self-supervised pretraining and fine-tuning. The diagnostic task used ECGs recorded within 14 days after symptom onset, regardless of reperfusion status. We conducted a four-phase reader study. Phases 1-3 compared unaided interpretation, a reference sheet, and additional AI assistance. In Phase 4, readers evaluated a new ECG set after randomization to reference-sheet support alone, with AI predictions, or with AI predictions plus gradient-weighted class activation mapping (Grad-CAM) heatmaps. Diagnostic correctness was analyzed using generalized linear mixed-effects models. Exploratory analyses examined differences by cardiologist status and experience. Results: Development and external-validation cohorts, including healthy controls, comprised 832 and 95 participants, respectively. For Takotsubo-STEMI discrimination, external-validation accuracy was 0.914, with areas under the receiver operating characteristic and precision-recall curves of 0.970 and 0.984. Seventy healthcare providers completed the reader study. The estimated probability of correct diagnosis increased from 0.632 without assistance to 0.864 with AI assistance (odds ratio [OR], 3.71; P<0.001). In randomized Phase 4, AI assistance improved accuracy vs reference-sheet support alone (OR, 4.28; 95% CI, 3.08-5.95; P<0.001), and Grad-CAM provided additional benefit (OR, 1.53; 95% CI, 1.04-2.25; P=0.033). Exploratory analyses showed greater improvement among less experienced readers; with AI assistance, non-cardiologists outperformed cardiologists. AI-assisted readers remained less accurate overall than the model alone. Conclusions: An ECG image-based ensemble differentiated Takotsubo syndrome from anterior STEMI and improved reader accuracy. Visual explanations provided additional benefit, while experience-related differences highlight the importance of effective human-AI interaction.