Search Results (2,573)

Nano- and pico-bubble water (NPB), containing hydrogen or ozone, is widely used as a cleaning agent due to its bactericidal and antiviral properties. However, some products—such as certain hydrogen waters—contain only large bubbles or have an extremely low bubble count, making them sometimes indistinguishable from ordinary drinking water. To accurately evaluate NPB activity, we developed a method for producing ultra-nano–pico-bubble water (NanoGAS water [NGW]), an ultra-fine bubble water that is stable and non-volatile over extended periods. By combining a mixed gas–liquid fluid rotary mixer and a shear filter, we produced ultra-fine bubbles that could be sealed in water. This method produced bubbles that remained stable in water even after 10 years since production. NGW has been clinically evaluated as a solvent for fecal microbiota transplantation (FMT) and has been demonstrated to be effective at improving bacterial engraftment in the intestinal tract in patients with autism spectrum disorder (ASD). Furthermore, encapsulating specific gases (hydrogen and ozone) can achieve more diverse effects. In this study, we evaluated the aroma-encapsulating effects, as well as the strength and persistence of the antimicrobial activity, of novel NGW formulations (Air-NGW, H₂-NGW, S-O₃-NGW, and L-O₃-NGW). Both H₂-NGW and O₃-NGW generated in this study demonstrated slight inclusion activity with volatile aromatic compounds (citral). Furthermore, both H₂-NGW (at ≥10% dilution) and O₃-NGW (even at a 1% dilution) exhibited sustained antibacterial efficacy against general viable bacteria for 24 weeks. Moreover, additive effects were observed when combined with antibacterial and antiviral compounds (polyoxometalates) developed by the authors. While further consideration, including cost-effectiveness, is needed to translate these findings into practical applications, they provide a fundamental framework for future research. Full article

Autism spectrum disorder (ASD) is genetically heterogeneous, involving rare and common variants that disrupt neurodevelopmental pathways. To explore this complexity, we performed whole-exome sequencing in children with ASD. Clinical phenotypes were systematically recorded, and severity was classified according to DSM-5 criteria. Variants were interpreted using ACMG guidelines, with recurrence analysis to identify genes shared across individuals and cohort enrichment testing against gnomAD. To examine genotype-phenotype relationships, we applied SKAT/SKAT-O across 16 phenotypes after covariate adjustment. Among 25 included individuals, pathogenic or likely pathogenic variants were found in 9, yielding a diagnostic yield of 36%. These involved genes linked to neurodevelopmental, epileptic, metabolic, and syndromic disorders. Recurrence analysis identified 586 genes present in at least two individuals, with PABPC1, GTF2I, PCLO, PKD1, and EP400 being the most frequent. SKAT/SKAT-O revealed the strongest burden associations for motor delay, aggressive behavior, mutism, anxiety, unresponsiveness to spoken voice, digestive disorder, and sleep disturbances, with limited overlap across phenotypes. Several recurrent genes also showed phenotype-specific associations. Overall, this integrative WES study provides clinically actionable diagnoses, highlights recurrent genes, and uncovers phenotype-specific signals, supporting convergent pathways with gene-level heterogeneity. Full article

Background/Objectives: Autism spectrum disorder (ASD) is a group of developmental disorders featured by social dysfunction, for which there still lacks effective treatments. Our previous study demonstrated that XAV939, a tankyrase inhibitor, could alleviate social dysfunction in two ASD mouse models via suppressing Wnt and glycolytic signaling. However, its further application is limited by poor brain–blood barrier penetration and low bioavailability. Methods: XAV939 was structurally optimized by esterification. The effects of XAV939-derivatives on Wnt/glycolysis were assessed by Western blotting, Topflash luciferase assay, lactate levels and the extracellular acidification rate. Social behaviors were evaluated by a three-chamber test, a resident–intruder test and ultrasonic vocalization. Biomimetic brain targeting was achieved by neuron-astrocyte hybrid cell membrane encapsulation. Periphery toxicities were examined by biochemical and histological analysis. Results: Three esterified XAV939 were synthesized. Data from both 293FT cells and primary Shank3b^-/- neurons revealed that an alkyl ester prodrug of XAV939 (named XAV939-L1) exhibited dual inhibition of Wnt/glycolysis. Intravenous injection of XAV939-L1 effectively improved the social function of Shank3b^-/- mice but showed hepatic side effects. Further, we made brain-targeting XAV939-L1 (XAV939-L1-BT) by neuron–astrocyte hybrid cell membrane encapsulation, which greatly enhanced the accumulation of XAV939-L1-BT in the brain and reduced its distribution in peripheral tissues (liver and intestine). At a half-dose of XAV939-L1, XAV939-L1-BT exhibited significant social improvement effects without obvious hepatic and intestinal toxicity. Conclusions: Our data demonstrated an esterified XAV939 and its biomimetic brain-targeted formula as a potential drug candidate for treating ASD-associated social dysfunction. Full article

Previous research has shown that children with autism spectrum disorder (ASD) often experience challenges related to sensory processing, spatial orientation, and navigation within built environments. Although environmental design factors have been recognized as important contributors to wayfinding performance, the specific role of interior color in supporting navigation and behavioral regulation remains insufficiently explored. This paper examines the effects of interior color on wayfinding performance in an autism healthcare facility. A mixed-methods design was used, involving experimental testing, behavioral observation, and questionnaire analysis. Twenty children with ASD were separated into control and experimental groups and exposed to color-modified environments as a result of a preliminary preference assessment. The success rate, time taken to navigate, and behavioral indicators were used to assess the performance. To determine differences between the color conditions, a statistical analysis was conducted. The findings indicate that cool colors, especially blue and green, are associated with improved wayfinding performance, including higher success rates, reduced navigation time, and calmer behavior. Conversely, warm colors like orange and pink were associated with poorer performance and greater distractibility. The findings indicated statistically significant differences between the tested color conditions, F(5, 14) = 5.87, p = 0.004, η²_p = 0.68. However, given the exploratory nature of the study and the limited sample size, these results should be interpreted cautiously. The study nevertheless provides preliminary evidence supporting the consideration of interior color as a potential design factor in autism-friendly environments. Full article

Background: Blue rubber bleb nevus syndrome (BRBNS) is a rare vascular disorder that primarily affects the skin and gastrointestinal tract, driven predominantly by somatic mosaic mutations, most commonly in TEK. Here, we report a case of a child with BRBNS and autism spectrum disorder (ASD). Case presentation: A 13-year-old boy diagnosed with ASD at 3 years presented with recurrent abdominal pain, blood in the stool, and severe anemia persisting for 6 months. At admission, his hemoglobin level was 5.6 g/dL. He had received a transfusion at age 6 for unexplained anemia. On examination, he appeared pale but stable, with bluish, compressible nodules on the right index finger and great toe, typical of BRBNS. Laboratory findings were consistent with chronic bleeding-induced iron deficiency. Endoscopic findings revealed multiple vascular lesions in the stomach, duodenum, ileum, and colon. Several colonic lesions were removed and pathologically confirmed as cavernous hemangiomas. Magnetic resonance enterography revealed additional small intestinal lesions. Whole-exome sequencing performed on buccal swab-derived DNA identified a heterozygous PDGFRA variant (c.2075G>T, p.Ser692Ile) classified as a variant of uncertain significance; no variants were identified in TEK, PIK3CA, or GNAQ. The patient underwent endoscopic resection of the larger lesions and received oral iron and a proton pump inhibitor. Hemoglobin stabilized at 11–12 g/dL, and no further transfusions were required. Conclusions: This case raises, but does not confirm, the possibility that genes other than TEK may contribute to BRBNS. The coexistence of ASD may be coincidental; a mechanistic link remains unproven. Careful endoscopic therapy and medical management controlled bleeding and anemia in this child. Full article

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental condition characterized by impairments in social communication, restricted and repetitive behaviors, sensory dysregulation, and frequent psychiatric comorbidities. Increasing attention has been directed toward acupuncture as a complementary neuro-modulatory intervention; however, its underlying molecular mechanisms remain incompletely understood. This review synthesizes evidence from preclinical, clinical, and molecular studies published between 2015 and 2025 to examine how acupuncture influences neurobiological pathways relevant to ASD. Current evidence indicates that acupuncture modulates multiple neurotransmitter systems, including glutamatergic, GABAergic, dopaminergic, serotonergic, and noradrenergic signaling, while also influencing neurotrophin-mediated plasticity, neuroinflammatory responses, and synaptic function. Studies conducted directly in ASD models demonstrate regulation of excitatory/inhibitory balance, monoaminergic signaling, neurotrophin pathways, and ASD-associated behavioral outcomes, whereas evidence from related neuropsychiatric conditions provides complementary mechanistic support for these pathways. Collectively, the findings suggest that acupuncture may act through coordinated modulation of interconnected neurotransmitter and neuroplasticity networks rather than a single molecular target. However, direct mechanistic evidence in ASD-specific models and clinical populations remains limited, and considerable heterogeneity exists among acupuncture protocols and outcome measures. Future studies integrating standardized stimulation paradigms with molecular, electrophysiological, neuroimaging, and behavioral assessments will be essential to validate the proposed mechanisms and clarify the translational potential of acupuncture in ASD. Full article

Background/Objectives: Fecal microbiota transplantation (FMT) is an established treatment for recurrent or refractory Clostridioides difficile infection (rCDI) and is increasingly investigated for other gastrointestinal and neuropsychiatric conditions in children. However, pediatric safety data remains limited. This study aimed to evaluate the short-term safety of colonoscopy-delivered FMT in a heterogeneous pediatric cohort. Methods: We conducted a retrospective single-centre cohort study of all FMT procedures performed at Vilnius University Hospital Santaros Klinikos between 2017 and 2025. Fecal material from three screened healthy pediatric donors was delivered to the caecum via colonoscopy under general anesthesia. Adverse events (AEs) arising within 72 h of each procedure were identified from the clinical records and assigned a severity grade using version 5.0 of the Common Terminology Criteria for Adverse Events (CTCAE). AEs were analyzed on a per-procedure basis. Associations between sex, age, and underlying indication and AE occurrence were assessed using Fisher’s exact test and the Mann–Whitney U test. Results: A total of 108 FMT procedures were performed on 73 children aged 2 to 17 years. The predominant indication was autism spectrum disorder (ASD) and other developmental disorders (80.6%). AEs were reported after 11 procedures (10.2%); all were classified as CTCAE grade 1 or 2, and no serious AEs occurred. The most common AEs were diarrhoea (5.6%) and vomiting (2.8%). In an exploratory comparison, procedures performed for ASD were associated with a lower AE rate compared with other indications combined (6.9% vs. 23.8%; OR 0.24, 95% CI 0.06–0.87). Conclusions: Colonoscopy-delivered FMT was well tolerated in pediatric patients, with a favorable short-term safety profile consistent with published pediatric FMT literature. As follow-up was largely passive and the cohort was predominantly children with ASD, the observed rate should be regarded as a lower bound, and these findings warrant confirmation in prospective studies. Full article

Early detection of autism spectrum disorder (ASD) in young children is essential for timely referral, developmental monitoring, and access to early intervention. However, conventional screening and diagnostic pathways often depend on parent-report instruments, episodic clinical observation, and specialist-administered assessments, which may delay identification during the first years of life. This scoping review maps the methodological landscape of early ASD detection from traditional clinical screening to multimodal artificial intelligence (AI). A structured literature search was conducted across major biomedical, psychological, and engineering databases for studies published between January 2010 and May 2026. After screening and eligibility assessment, 65 evidence sources were included in the qualitative synthesis, with additional methodological guidelines used to support reporting and appraisal. The reviewed evidence shows that early ASD detection is increasingly shifting from single-session clinical assessment toward multidimensional risk characterization. Clinical and behavioral screening tools remain the foundation of early identification, while eye tracking, video-based motor analysis, acoustic and vocal biomarkers, electroencephalography (EEG), functional near-infrared spectroscopy (fNIRS), and molecular or genomic indicators provide complementary information across different developmental windows. AI-based methods, including machine learning, deep learning, Transformer architectures, multimodal fusion strategies, and foundation-model-based representation learning, may improve the objective quantification of gaze, movement, vocalization, neural activity, and biological risk. Nevertheless, most AI-assisted systems remain limited by small and heterogeneous datasets, insufficient external validation, population bias, privacy concerns, computational burden, and limited interpretability. This review argues that future early ASD detection systems should be developed as clinician-supervised decision-support tools rather than autonomous diagnostic instruments. Clinically meaningful progress will require robust external validation, privacy-preserving deployment, age-appropriate risk stratification, and intrinsically interpretable architectures that align model outputs with developmental and clinical knowledge. Full article

The present pilot study examined the feasibility, acceptability, and preliminary effectiveness of a VR-based social skills intervention for children with Autism Spectrum Disorder (ASD). Five participants aged 7–17 years participated in a six-session intervention protocol, with each session lasting approximately 45 min. The study employed a randomized AB/BA crossover design, in which participants completed two sessions under an Active condition involving instructional prompts, corrective feedback, and positive reinforcement, and two sessions under a Neutral condition without prompts or reinforcement. Following the intervention phases, participants completed a Generalization session in a novel virtual environment and a Follow-up session to assess skill retention over time. The intervention incorporated two interactive VR scenarios: (a) a simulated classroom environment, where a teacher-avatar engaged participants in structured question-and-answer activities, and (b) a virtual school playground, where participants interacted with peer avatars in social communication tasks designed to promote conversational engagement, turn-taking, and social reciprocity. All avatar interactions and trial progressions were controlled by the therapist through a dedicated monitoring interface, ensuring standardized implementation across participants. Evaluation was based on qualitative observation, analysis of recorded video material, and brief post-intervention parent interviews. Preliminary findings suggest that the intervention was feasible, well accepted by participants, and associated with improvements in social communication performance, supporting the potential of VR as a complementary tool for social skills training in children with ASD. Full article

Background: Although genetic-environmental interactions are established in autism spectrum disorder (ASD), how environmental toxicants confer susceptibility remains unclear. This study aimed to investigate potential relationship between genetically predicted cerebrospinal fluid (CSF), metabolite levels and ASD liability, and to prioritize regulatory genes, key pathways, and candidate environmental toxicants. Methods: Using two ASD GWAS datasets (exploration data: 18,381 ASD cases/27,969 controls; validation data: 18,235 ASD cases/36,741 controls), we applied multi-omics approaches to prioritize ASD-associated CSF metabolites, regulatory SNPs, and genes. Enrichment analysis and protein–protein interaction (PPI) network analysis were performed on these metabolite-related genes to explore the potential mechanisms linking CSF metabolic disturbances to ASD. Finally, candidate environmental neurotoxicants were screened through protein-chemical interaction analysis, with binding relationships assessed via molecular docking prediction. Results: Two-sample Mendelian randomization (MR) analysis prioritized adenine and proline as candidate CSF metabolites with potential risk associations with ASD. Summary-data-based MR (SMR) prioritized 39 brain-specific quantitative trait loci (QTL) involving 35 candidate regulatory genes, including dual-metabolite modulator GRM8. Functional enrichment analyses suggested potential associations with mitochondrial dysfunction, Hippo signaling pathway, and microtubule dynamics impairment, with protein–protein interaction networks highlighting KATNA1/KATNAL2 as hubs. Protein-chemical interaction screening nominated 14 candidate environmental toxicants, including established chemicals (acetaminophen, valproic acid, estradiol) and novel candidates (SB-431542, K 7174, benzo[a]pyrene), with docking affinity assessed computationally. Conclusions: Our study provides suggestive evidence that elevated adenine and proline may be potential risk factors for ASD and suggests possible involvement of the mitochondrial–Hippo–microtubule pathway. We also propose benzo[a]pyrene as a candidate environmental toxicant that may perturb CSF metabolism. However, given the limited statistical significance, these findings require further validation. Full article

Background: Digital behavioral phenotyping of autism spectrum disorder (ASD) offers a promising approach for developing more scalable diagnostic frameworks across diverse global contexts. Machine learning (ML) models show promise for ASD diagnosis using behavioral videos, but critical questions remain regarding whether models trained on data from one country work in another, and how the background of the raters affects the accuracy. Our work addresses these questions by testing whether ML models can accurately diagnose ASD across different populations and rater groups. Methods: This work evaluates the performance of a supervised ML framework for binary classification of ASD versus non-ASD [speech, language and communication disorders (SLC) + neurotypical (NT)] in a cohort of 227 children in Bangladesh. We first assessed the cross-domain model transferability of a clinical-instrument-trained logistic regression model (LR-9) on behavioral ratings that were based on videos of Bangladeshi children interacting with caregivers and toys at two major child development centers in Dhaka, Bangladesh. We then trained five diverse classifiers (Logistic Regression, Random Forest, XGBoost, SVM, and RuleFit) on the full annotated Bangladeshi dataset. Using SHAP-based consensus elbow feature selection, we identified a compact set of features that maintained the performance. Finally, we developed ensemble models to improve predictive stability. Results: The LR-9 model, originally trained on U.S. clinical instrument data, was evaluated on video-based behavioral ratings from 214 Bangladeshi children. When tested on Bangladeshi clinician ratings, the LR-9 model achieved a sensitivity of 86.1% (95% CI: [0.78–0.93]) and AUC of 0.79 (95% CI: [0.73–0.86]). The distinction across rater groups was between trained raters (clinicians and students) and crowd workers, who showed lower sensitivity 28.5% (95% CI: [0.21, 0.39]). When tested on the aggregated ratings from all groups, the model achieved an AUC of 0.78 (95% CI: [0.72–0.84]). Inter-rater reliability followed the same pattern: individual agreement was fair (Krippendorff’s α = 0.26), but the multi-rater consensus was reliable (ICC(1,k) = 0.84), with Bangladeshi clinicians showing the highest agreement (α = 0.34) and crowd workers the lowest (α = 0.20). We then trained new models directly on the Bangladeshi ratings. All model types achieved similar AUC values (0.86–0.89), with overlapping confidence intervals. Using just 8–11 key behaviors kept the similar performance while cutting the features by 66–75%. Combining ensembles gave similar results (e.g., Bayesian averaging: AUC 0.88 [0.78, 0.95]) but with more stable predictions. Conclusion: This study provides evidence that mobile video-based ASD diagnosis can achieve comparable performance (AUC: 0.89 [0.76, 0.96]) to models trained on clinical instrument data. This work contributes to the development of broader adaptable autism detection tools, bypassing the dependence on traditional clinical instrument data. Full article

Background: Neurodevelopmental disorders, including Autism Spectrum Disorder (ASD), Attention-Deficit/Hyperactivity Disorder (ADHD), and Specific Learning Disorder (SLD), as well as neuromuscular conditions such as Duchenne Muscular Dystrophy (DMD), are frequently associated with emotional and behavioral difficulties that may affect children’s functioning and family well-being. However, direct comparisons across these heterogeneous clinical populations remain limited. This study aimed to compare emotional and behavioral profiles across children and adolescents with ASD, ADHD, SLD, and DMD using the Child Behavior Checklist (CBCL/6–18). Methods: This cross-sectional comparative study included 197 children and adolescents (ASD: 47; SLD: 50; ADHD: 50; DMD: 50) assessed at the Clinical Psychology Unit of the Fondazione Policlinico Universitario Agostino Gemelli IRCCS (Rome, Italy) between 2017 and 2024. Emotional and behavioral functioning was evaluated using the parent-report CBCL/6–18. Group differences were examined using one-way ANOVA or Welch’s ANOVA, as appropriate, followed by Tukey’s HSD or Games–Howell post hoc comparisons. Results: Significant group differences emerged for Internalizing Problems, Externalizing Problems, and Total Problems (all p < 0.001). ADHD showed the highest overall emotional and behavioral burden, with 76% of participants scoring in the clinical range for both Internalizing and Total Problems and 60% for Externalizing Problems. Elevated internalizing difficulties characterized ASD and SLD, whereas children with DMD showed generally lower CBCL scores, although 26% fell within the clinical range for Internalizing Problems. Effect sizes were moderate to large (η² = 0.168–0.273), supporting the presence of both shared and disorder-specific emotional and behavioral patterns across heterogeneous developmental conditions. Conclusions: The findings highlight the heterogeneity of emotional–behavioral profiles across neurodevelopmental and neuromuscular conditions. The use of a common standardized assessment framework enabled the identification of both shared and disorder-specific patterns, supporting the potential value of a transdiagnostic approach to individualized assessment. Future longitudinal and multi-informant studies are warranted to clarify developmental trajectories and improve the identification of clinically meaningful emotional and behavioral needs across heterogeneous pediatric populations. Full article

Background: Duchenne muscular dystrophy (DMD) is an X-linked disorder caused by out-of-frame variants in the DMD gene, resulting in dystrophin deficiency and progressive muscle degeneration. Beyond motor involvement, evidence links DMD to cognitive impairment and an emotional–behavioral (EB) burden, potentially related to the altered expression of brain dystrophin isoforms (Dp71, Dp140, Dp427). Objectives: To screen for major neurodevelopmental, cognitive and/or EB difficulties in a monocentric cohort of children with DMD. Methods: This cross-sectional study included 21 children with DMD. Neuropsychiatric difficulties were assessed using a multimodal psychometric battery. Cognitive, neurodevelopmental and genetic data were retrospectively collected and analyzed. Results: In the cohort, attention-deficit/hyperactivity disorder (ADHD)-related findings were predominantly inattentive, with 3/20 children (15.0%) scoring within the clinical range on at least one inattention subscale. For measures assessing autism spectrum disorder (ASD)-related features, scores above the normative cutoff emerged in 7/20 children (35.0%), while only 2/20 (10.0%) scored within the clinical range. Internalizing problems represented the predominant EB difficulties, and emotional dysregulation (ED) emerged as a plausible area of vulnerability in the cohort. Among the 11 of 21 children with available Full-Scale Intelligence Quotient (FSIQ) data, five (45.5%) had an FSIQ below 85. Of these, four of five (80.0%) presented the predicted Dp140−/Dp71+ brain dystrophin isoform expression pattern, whereas one of five (20.0%) presented the Dp140+/Dp71+ pattern. Conclusions: Children with DMD showed heterogeneous neuropsychiatric and cognitive features in the employed screening battery. These preliminary findings, if confirmed in larger cohorts, support the potential of a broader neuropsychiatric screening assessment to optimize care pathways. Full article

Functional near-infrared spectroscopy (fNIRS) is a portable, non-invasive tool for studying cortical function in children with neurodevelopmental and neurological disorders. Although fNIRS use is increasing, heterogeneous study paradigms and cortical targets have limited cross-condition comparisons and the identification of shared research priorities. This systematic review maps cortical regions and reporting patterns in five key conditions—autism spectrum disorder (ASD), attention-deficit/hyperactivity disorder (ADHD), cerebral palsy (CP), hypoxic–ischemic encephalopathy (HIE), and epilepsy (Ep)—from January 2015 to December 2025. A systematic search across five databases identified 72 relevant studies meeting PRISMA 2020 criteria, revealing both similarities and differences across disorders. The prefrontal cortex (PFC) was studied in all five conditions (5/5: 100%), making it the most consistently investigated cortical region. The parietal and temporal cortices were studied in 4 of 5 conditions (80%). The frontal cortex had the most regions investigated, while the temporal cortex showed the most consistent coverage across conditions (2.25 conditions per region). Beyond regional preferences, condition-specific patterns were aligned with their disorder phenotypes: social brain networks in ASD, prefrontal executive systems in ADHD, sensorimotor changes in CP, cerebrovascular monitoring in HIE, and state-dependent changes in Ep. Across conditions, researchers found altered prefrontal activity, disrupted connectivity, and compensatory brain responses, supporting broader frameworks. Collectively, these findings identify the PFC as a shared target for transdiagnostic fNIRS investigations while highlighting important gaps in regional coverage and methodological consistency. Despite methodological differences, fNIRS shows promise for identifying both shared and unique brain patterns in pediatric neurodevelopmental and neurological disorders. This review provides a framework for prioritizing cortical targets and guiding future standardized fNIRS research in pediatric populations. Full article

Background/Objectives: Gut dysbiosis in children with autism spectrum disorder (ASD) has been associated with alterations in microbial ecology and metabolic function that may contribute to gastrointestinal dysfunction and the severity of clinical manifestations. Synbiotic and probiotic supplementation has emerged as a promising microbiome-targeted strategy for ASD; however, its effects on gut microbiome composition, functional potential, and clinical outcomes remain incompletely understood. We conducted a longitudinal study of Mexican children diagnosed with ASD to analyze changes in the composition, diversity, and functional potential of the gut microbiome during six months of multi-strain synbiotic supplementation. Methods: Stool samples were collected from 25 children with ASD at baseline and after 3 and 6 months of multi-strain synbiotic supplementation. Gut microbiome composition and diversity were analyzed by 16S rRNA gene sequencing, whereas whole metagenome sequencing (WMS) was performed in a subset of samples to evaluate the functional potential of the fecal microbiome. Gastrointestinal symptoms were assessed using the Rome IV criteria, and ASD severity was evaluated with the Childhood Autism Rating Scale (CARS). Results: Twenty-five children with ASD completed the 6 months of synbiotic supplementation. Overall, ASD severity decreased, reflected by a reduction in total CARS score, and improvements in several CARS domains. Gastrointestinal symptoms also decreased significantly. Longitudinal microbiome profiling revealed significant taxonomic and diversity changes over the supplementation period, while WMS identified changes in microbial metabolic potential, including enrichment of tryptophan biosynthesis pathways and reduced L-rhamnose degradation. Conclusions: This exploratory research provides proof-of-concept evidence supporting multi-strain synbiotic supplementation in children with ASD. Larger controlled studies are needed to confirm these findings and clarify their relevance to microbiota–gut–brain axis interactions. The observed concordance between clinical improvements and microbiome remodeling supports further investigation of microbiome-targeted interventions according to ASD severity and duration of supplementation. Full article