Barbara Rašková

Research on autism spectrum disorders (ASDs) has so far focused primarily on the gut–brain axis, whereas the role of the gut–liver axis remains insufficiently explored. The aim of this study was to evaluate whether microbiota derived from girls with ASD induces dysbiotic changes in gut microbiota composition and leads to alterations in gut–liver axis processes in pseudo-germ-free (PGF) BALB/c mice. We also examined whether these processes could be modulated by altering the gut microbiota using the probiotic strains Lactiplantibacillus plantarum CCM 7512 and Limosilactobacillus reuteri CCM 8617 in combination with ground flaxseed (Linum usitatissimum L.) as a source of omega-3 polyunsaturated fatty acids and fermentable fiber. Colonization with fecal microbiota derived from girls with ASD resulted in dysbiotic changes in the composition of the cecal microbiota, characterized by an increased relative abundance of Escherichia–Shigella, Fusobacterium, Alistipes, and the Ruminococcus gnavus group. These changes were accompanied by impaired intestinal mucosal integrity, altered metabolomic pathways related mainly to aromatic amino acids and lipid metabolism, increased hepatic immunoreactivity of iNOS and COX-2, and elevated activity of the liver-specific LDH-5 isoenzyme. These results suggest that synbiotic intervention contributed to remodeling of the cecal microbiota composition, restoration of intestinal epithelial integrity, and modulation of metabolomic pathways, which was reflected by reduced immunoreactivity of iNOS and COX-2 in liver tissue and decreased activity of the LDH-5 isoenzyme. These findings support the role of microbiota-mediated metabolic processes in communication between the gut and the liver within the gut–liver axis.

Background: Autism spectrum disorder (ASD) is a heterogeneous group of neurodevelopmental disorders characterized by a complex, multifactorial etiology with a strong genetic contribution. Our study aimed to evaluate the link between the burden of rare genetic variants within a specific panel of ASD and intellectual disability-associated genes and phenotypic variability in a cohort of children with autism in Slovakia. Methods: Gene burden scores were calculated based on pathogenic, likely pathogenic, and uncertain significance rare DNA variants identified by whole-exome sequencing. We then assessed the effect of three different scoring methods on the variance across 15 psycho-behavioral parameters describing the phenotypic profiles of 117 ASD probands. Results: The burden score showed a significant multivariate effect on the combination of psycho-behavioral parameters. This score was associated with the social affect of ADOS-2, as well as with the socialization domain, and total adaptive behavior scores from the Vineland Adaptive Behavior Scales-3 (VABS). While a score based solely on count of pathogenic and likely pathogenic variants did not show a multivariate effect, incorporating variants of uncertain significance revealed a multivariate effect on two adaptive behavior parameters: daily living skills and total adaptive behavior score (VABS). Conclusions: Our findings partially explain the variability in phenotypic manifestation in our ASD patient cohort, highlighting the importance of considering the cumulative effect of rare genetic variants, including those of uncertain significance, in shaping the diverse clinical presentation of ASD.

The increasing incidence of autism spectrum disorder (ASD) increases the urgency of establishing the mechanism of its development for effective prevention and treatment. ASD’s etiology includes genetic predisposition and environmental triggers, both of which can play a role in the changed microbiota. Recent research has proved the impact of maternal microbiota on the neurodevelopment of the child. To investigate the co-play of genetic and microbiota factors in ASD development, we performed fecal microbiota transplantation (FMT) from children with ASD to female Shank3b^+/− mice and studied the autism-like symptoms in the male Shank3b^−/− and wild-type (WT) offspring. WT animals with prenatal exposure to ASD microbiota had delayed neurodevelopment and impaired food intake behavior, but also elevated plasma leptin concentration and body weight. Shank3b^−/− mice after FMT ASD exhibited impaired learning and exacerbated anxiety-like behavior in adulthood. Interestingly, FMT ASD improved learning in adolescent Shank3b^−/− mice. Prenatal exposure to ASD microbiota decreased the activity of hypocretin neurons of the lateral hypothalamic area in both genotypes. The combination of genetic predisposition and FMT ASD led to an increased colon permeability, evaluated by zonula occludens (ZO1, ZO3) and claudin factors. These results suggest the effect of parental FMT exposure on shaping offspring behavior in Shank3b^−/− mice and the potential of microbiota in the modulation of ASD.

Children with Autism Spectrum Disorder (ASD) differ in adaptive functioning, executive functioning, and the distribution of intellectual abilities compared to their typically developing (TD) peers. Research consistently demonstrates a link between executive functions and various aspects of adaptive behavior. This study aims to compare cognitive abilities, executive functions, and adaptive behavior in non- or minimally verbal preschool-aged (3–6 years) children with ASD (n = 53) and TD peers (n = 79). The study examines broad cognitive abilities and executive functions as predictors of adaptive behavior. For this purpose, the Vineland Adaptive Behavior Scales, Third Edition, the Behavior Rating Inventory of Executive Functioning–Preschool Version, and the SON-R 2½-7 intelligence test were utilized. The results indicate that non- or minimally verbal preschoolers with ASD exhibit significant executive difficulties, particularly in working memory, and show marked differences in cognitive abilities compared to TD controls. Significant group differences were observed across all adaptive behavior domains, with the largest effect on Socialization and the smallest on Daily Living Skills. In the ASD group, executive functions and cognitive domains accounted for a substantial portion of the variance in the Communication domain, with slightly less influence observed in Daily Living Skills and Socialization. In contrast, the significant role of cognitive abilities and executive difficulties as predictors of adaptive behavior in everyday functioning was not confirmed for TD children. These findings highlight the critical role of executive functioning as a predictor of adaptive behavior in preschool children with ASD.

Background: Multiple research teams have documented various abnormalities in erythrocyte properties in children with autism spectrum disorder (ASD) compared with neurotypical individuals. Reduced erythrocyte deformability, a crucial factor for microcirculation and oxygen delivery, may affect brain function. Other key factors like nitric oxide (NO) and Na,K-ATPase-regulated cation transport also play roles in both erythrocyte deformability and ASD, suggesting a possible relationship between erythrocyte parameters and autism severity. Thus, this study aims to describe these associations, exploring erythrocyte properties as potential biomarkers in ASD. Methods: A total of 179 ASD children were enrolled in this study. Diagnosis was confirmed by the Autism Diagnostic Observation Schedule—Second Edition (ADOS-2) and Autism Diagnostic Interview-Revised. The Vineland Adaptive Behavior Scales, Third Edition (VABS-3), was used to assess adaptive behavior. RBC deformability was measured using a filtration technique, while NO production by RBCs was assessed via DAF-2DA fluorescence. Na,K-ATPase kinetics and RBC osmotic resistance were evaluated spectrophotometrically. Results: Children with more severe ASD symptoms had more impaired deformability and osmotic resistance than children with mild symptoms. Higher RBC NO production was linked to better scores in some VABS-3 subdomains, and in the social affect domain of ADOS-2. Higher affinity of Na,K-ATPase for sodium negatively correlated with the occurrence of repetitive and restricted behavior—one of the core ASD symptoms. Conclusions: This study identified potential links between ASD severity and RBC properties. While erythrocyte quality can influence ASD symptomatology, the observed relationships—such as those involving RBC deformability, NO production, Na,K-ATPase kinetics, and osmotic resistance—were not strong or consistent enough to be considered reliable diagnostic or prognostic biomarkers.

The main goal of our research was to monitor changes in the mental health of Slovak families with children with autism spectrum disorder (ASD) compared to families with neurotypical children during three waves of the COVID-19 pandemic. We focused on the prevalence of depression, anxiety, stress and different stressors of parents. In children, we explored maladaptive behavior and the availability of interventions for children with ASD. The data were collected using an extensive questionnaire including the Depression, Anxiety, and Stress Scale-42 questionnaire (DASS-42) and two subscales of the Vineland Adaptive Behavior Scales (VABS-3). The research sample consisted of a total of 506 parents, 236 of whom have a child with ASD. Parents of children with ASD reported elevated anxiety during the first wave, while changes were found in parents of neurotypical children. During the second wave, the prevalence of anxiety, depression and stress experienced by parents in both groups increased, but significantly more in parents with ASD children. The internalizing maladaptive behavior of children with ASD also increased. During the third wave, no significant differences between the groups of parents were found in stress and anxiety, but parents of ASD children scored higher in depression. Externalized maladaptive behavior of neurotypical children increased, with minimal changes in children with ASD, which can be explained by the improved therapy availability for children with ASD, also observed in our study.