Bridging Genetic Insights with Neuroimaging in Autism Spectrum Disorder-A Systematic Review.

Autism Spectrum Disorder (ASD) is an early onset neurodevelopmental disorder characterized by impaired social interaction and communication, and repetitive patterns of behavior. Family studies show that ASD is highly heritable, and hundreds of genes have previously been implicated in the disorder; however, the etiology is still not fully clear. Brain imaging and electroencephalography (EEG) are key techniques that study alterations in brain structure and function. Combined with genetic analysis, these techniques have the potential to help in the clarification of the neurobiological mechanisms contributing to ASD and help in defining novel therapeutic targets. To further understand what is known today regarding the impact of genetic variants in the brain alterations observed in individuals with ASD, a systematic review was carried out using Pubmed and EBSCO databases and following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. This review shows that specific genetic variants and altered patterns of gene expression in individuals with ASD may have an effect on brain circuits associated with face processing and social cognition, and contribute to excitation-inhibition imbalances and to anomalies in brain volumes.

Identification of Neurotransmission and Synaptic Biological Processes Disrupted in Autism Spectrum Disorder Using Interaction Networks and Community Detection Analysis.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by communication deficits and repetitive behavioral patterns. Hundreds of candidate genes have been implicated in ASD, including neurotransmission and synaptic (NS) genes; however, the genetic architecture of this disease is far from clear. In this study, we seek to clarify the biological processes affected by NS gene variants identified in individuals with ASD and the global networks that link those processes together. For a curated list of 1216 NS candidate genes, identified in multiple databases and the literature, we searched for ultra-rare (UR) loss-of-function (LoF) variants in the whole-exome sequencing dataset from the Autism Sequencing Consortium (N = 3938 cases). Filtering for population frequency was carried out using gnomAD (N = 60,146 controls). NS genes with UR LoF variants were used to construct a network of protein-protein interactions, and the network's biological communities were identified by applying the Leiden algorithm. We further explored the expression enrichment of network genes in specific brain regions. We identified 356 variants in 208 genes, with a preponderance of UR LoF variants in the PDE11A and SYTL3 genes. Expression enrichment analysis highlighted several subcortical structures, particularly the basal ganglia. The interaction network defined seven network communities, clustering synaptic and neurotransmitter pathways with several ubiquitous processes that occur in multiple organs and systems. This approach also uncovered biological pathways that are not usually associated with ASD, such as brain cytochromes P450 and brain mitochondrial metabolism. Overall, the community analysis suggests that ASD involves the disruption of synaptic and neurotransmitter pathways but also ubiquitous, but less frequently implicated, biological processes.

Antimicrobial Effects and Antioxidant Activity of Myrtus communis L. Essential Oil in Beef Stored under Different Packaging Conditions.

The aim of this study was to assess the antimicrobial effects of myrtle (Myrtus communis L.) essential oil (EO) on pathogenic (E. coli O157:H7 NCTC 12900; Listeria monocytogenes ATCC BAA-679) and spoilage microbiota in beef and determine its minimum inhibitory concentration (MIC) and antioxidant activity. The behavior of LAB, Enterobacteriaceae, Pseudomonas spp., and fungi, as well as total mesophilic (TM) and total psychotropic (TP) counts, in beef samples, was analyzed during storage at 2 and 8 °C in two different packaging systems (aerobiosis and vacuum). Leaves of myrtle were dried, its EO was extracted by hydrodistillation using a Clevenger-type apparatus, and the chemical composition was determined using chromatographical techniques. The major compounds obtained were myrtenyl acetate (15.5%), ß-linalool (12.3%), 1,8-cineole (eucalyptol; 9.9%), geranyl acetate (7.4%), limonene (6.2%), α-pinene (4.4%), linalyl o-aminobenzoate (5.8%), α-terpineol (2.7%), and myrtenol (1.2%). Myrtle EO presented a MIC of 25 µL/mL for E. coli O157:H7 NCTC 12900, E. coli, Listeria monocytogenes ATCC BAA-679, Enterobacteriaceae, and E. coli O157:H7 ATCC 35150 and 50µL/mL for Pseudomonas spp. The samples packed in aerobiosis had higher counts of deteriorative microorganisms than samples packed under vacuum, and samples with myrtle EO presented the lowest microbial contents, indicating good antimicrobial activity in beef samples. Myrtle EO is a viable natural alternative to eliminate or reduce the pathogenic and deteriorative microorganisms of meat, preventing their growth and enhancing meat safety.

Disease similarity network analysis of Autism Spectrum Disorder and comorbid brain disorders.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder with heterogeneous clinical presentation, variable severity, and multiple comorbidities. A complex underlying genetic architecture matches the clinical heterogeneity, and evidence indicates that several co-occurring brain disorders share a genetic component with ASD. In this study, we established a genetic similarity disease network approach to explore the shared genetics between ASD and frequent comorbid brain diseases (and subtypes), namely Intellectual Disability, Attention-Deficit/Hyperactivity Disorder, and Epilepsy, as well as other rarely co-occurring neuropsychiatric conditions in the Schizophrenia and Bipolar Disease spectrum. Using sets of disease-associated genes curated by the DisGeNET database, disease genetic similarity was estimated from the Jaccard coefficient between disease pairs, and the Leiden detection algorithm was used to identify network disease communities and define shared biological pathways. We identified a heterogeneous brain disease community that is genetically more similar to ASD, and that includes Epilepsy, Bipolar Disorder, Attention-Deficit/Hyperactivity Disorder combined type, and some disorders in the Schizophrenia Spectrum. To identify loss-of-function rare de novo variants within shared genes underlying the disease communities, we analyzed a large ASD whole-genome sequencing dataset, showing that ASD shares genes with multiple brain disorders from other, less genetically similar, communities. Some genes (e.g., SHANK3, ASH1L, SCN2A, CHD2, and MECP2) were previously implicated in ASD and these disorders. This approach enabled further clarification of genetic sharing between ASD and brain disorders, with a finer granularity in disease classification and multi-level evidence from DisGeNET. Understanding genetic sharing across disorders has important implications for disease nosology, pathophysiology, and personalized treatment.

A Role for Gene-Environment Interactions in Autism Spectrum Disorder Is Supported by Variants in Genes Regulating the Effects of Exposure to Xenobiotics.

Heritability estimates support the contribution of genetics and the environment to the etiology of Autism Spectrum Disorder (ASD), but a role for gene-environment interactions is insufficiently explored. Genes involved in detoxification pathways and physiological permeability barriers (e.g., blood-brain barrier, placenta and respiratory airways), which regulate the effects of exposure to xenobiotics during early stages of neurodevelopment when the immature brain is extremely vulnerable, may be particularly relevant in this context. Our objective was to identify genes involved in the regulation of xenobiotic detoxification or the function of physiological barriers (the XenoReg genes) presenting predicted damaging variants in subjects with ASD, and to understand their interaction patterns with ubiquitous xenobiotics previously implicated in this disorder. We defined a panel of 519 XenoReg genes through literature review and database queries. Large ASD datasets were inspected for in silico predicted damaging Single Nucleotide Variants (SNVs) (N = 2,674 subjects) or Copy Number Variants (CNVs) (N = 3,570 subjects) in XenoReg genes. We queried the Comparative Toxicogenomics Database (CTD) to identify interaction pairs between XenoReg genes and xenobiotics. The interrogation of ASD datasets for variants in the XenoReg gene panel identified 77 genes with high evidence for a role in ASD, according to pre-specified prioritization criteria. These include 47 genes encoding detoxification enzymes and 30 genes encoding proteins involved in physiological barrier function, among which 15 are previous reported candidates for ASD. The CTD query revealed 397 gene-environment interaction pairs between these XenoReg genes and 80% (48/60) of the analyzed xenobiotics. The top interacting genes and xenobiotics were, respectively, CYP1A2, ABCB1, ABCG2, GSTM1, and CYP2D6 and benzo-(a)-pyrene, valproic acid, bisphenol A, particulate matter, methylmercury, and perfluorinated compounds. Individuals carrying predicted damaging variants in high evidence XenoReg genes are likely to have less efficient detoxification systems or impaired physiological barriers. They can therefore be particularly susceptible to early life exposure to ubiquitous xenobiotics, which elicit neuropathological mechanisms in the immature brain, such as epigenetic changes, oxidative stress, neuroinflammation, hypoxic damage, and endocrine disruption. As exposure to environmental factors may be mitigated for individuals with risk variants, this work provides new perspectives to personalized prevention and health management policies for ASD.

Gene Variants Involved in Nonsense-Mediated mRNA Decay Suggest a Role in Autism Spectrum Disorder.

Autism Spectrum Disorder (ASD) is a heterogeneous neurodevelopmental condition with unclear etiology. Many genes have been associated with ASD risk, but the underlying mechanisms are still poorly understood. An important post-transcriptional regulatory mechanism that plays an essential role during neurodevelopment, the Nonsense-Mediated mRNA Decay (NMD) pathway, may contribute to ASD risk. In this study, we gathered a list of 46 NMD factors and regulators and investigated the role of genetic variants in these genes in ASD. By conducting a comprehensive search for Single Nucleotide Variants (SNVs) in NMD genes using Whole Exome Sequencing data from 1828 ASD patients, we identified 270 SNVs predicted to be damaging in 28.7% of the population. We also analyzed Copy Number Variants (CNVs) from two cohorts of ASD patients (N = 3570) and discovered 38 CNVs in 1% of cases. Importantly, we discovered 136 genetic variants (125 SNVs and 11 CNVs) in 258 ASD patients that were located within protein domains required for NMD. These gene variants are classified as damaging using in silico prediction tools, and therefore may interfere with proper NMD function in ASD. The discovery of NMD genes as candidates for ASD in large patient genomic datasets provides evidence supporting the involvement of the NMD pathway in ASD pathophysiology.

Identification of biological mechanisms underlying a multidimensional ASD phenotype using machine learning.

The complex genetic architecture of Autism Spectrum Disorder (ASD) and its heterogeneous phenotype makes molecular diagnosis and patient prognosis challenging tasks. To establish more precise genotype-phenotype correlations in ASD, we developed a novel machine-learning integrative approach, which seeks to delineate associations between patients' clinical profiles and disrupted biological processes, inferred from their copy number variants (CNVs) that span brain genes. Clustering analysis of the relevant clinical measures from 2446 ASD cases in the Autism Genome Project identified two distinct phenotypic subgroups. Patients in these clusters differed significantly in ADOS-defined severity, adaptive behavior profiles, intellectual ability, and verbal status, the latter contributing the most for cluster stability and cohesion. Functional enrichment analysis of brain genes disrupted by CNVs in these ASD cases identified 15 statistically significant biological processes, including cell adhesion, neural development, cognition, and polyubiquitination, in line with previous ASD findings. A Naive Bayes classifier, generated to predict the ASD phenotypic clusters from disrupted biological processes, achieved predictions with a high precision (0.82) but low recall (0.39), for a subset of patients with higher biological Information Content scores. This study shows that milder and more severe clinical presentations can have distinct underlying biological mechanisms. It further highlights how machine-learning approaches can reduce clinical heterogeneity by using multidimensional clinical measures, and establishes genotype-phenotype correlations in ASD. However, predictions are strongly dependent on patient's information content. Findings are therefore a first step toward the translation of genetic information into clinically useful applications, and emphasize the need for larger datasets with very complete clinical and biological information.

Efficacy of a public promotion program on children's oral health / Eficácia de um programa público de promoção de saúde bucal em crianças

Abstract Objective: To assess the efficacy of the Baby's Mouth early dental care prevention and promotion program in preventing oral diseases (caries, gingivitis, or malocclusions) in children attended since 2010. Methods: This was a cross-sectional and cohort study that assessed 252 children between 36 and 60 months of age in both sexes. The children were divided into three groups: G1: effective participants of the program from birth; G2: children who have stopped participating for more than 24 months, and G3: children who have never attended a prevention program. The evaluation was carried out in two stages: first, an interview with the mothers and, afterwards, a clinical children examination to assess the presence of caries, gingivitis, and malocclusion. The chi-squared test was used for statistical analysis between groups (p < 0.05). Results: The diseases assessed were: caries (G1: 5.9%, G2: 54.7%, G3: 70%), gingivitis (G1: 8.3%, G2: 17.9%, G3: 40.5%), and malocclusion (G1: 22.6%; G2: 28.6%; G3: 50%). For gingivitis, there was no significant difference when comparing G1 and G2 (p = 0.107), but it was significant between G1 and G3 (p < 0.001). Regarding malocclusion, a statistically significant relationship was observed (p = 0.004) among all groups. Conclusion: The prevention and promotion program in public oral health was effective in preventing caries disease, gingivitis, and malocclusion in children under 5 years of age.

Resumo Objetivo: Avaliar a eficácia do programa de prevenção e promoção de cuidados dentários precoce da boca do bebê, a fim de prevenir doenças bucais (cáries, gengivite ou má oclusões) em crianças atendidas desde 2010. Métodos: Estudo transversal e de coorte com avaliação de 252 crianças entre 36 e 60 meses de idade de ambos os sexos. As crianças foram divididas em dois grupos: G1: participantes efetivos do programa a partir do nascimento; G2: crianças que pararam de participar por mais de 24 meses do programa; e G3: crianças que nunca participaram de um programa de prevenção. A avaliação foi feita em dois estágios: entrevista com as mães e, depois, um exame clínico nas crianças para analisar cáries, gengivite e oclusão. Foi utilizado o teste qui-quadrado para análise estatística entre os grupos (p < 0,05). Resultados: As doenças analisadas foram: cáries (G1: 5,9%, G2: 54,7%, G3: 70%), gengivite (G1: 8,3%, G2: 17,9%, G3: 40,5%) e má oclusão (G1: 22,6%; G2: 28,6%; G3: 50%). Para gengivite, não houve diferença significativa ao comparar G1 e G2 (p = 0,107), porém a diferença foi extremamente significativa entre G1 e G3 (p < 0,001). Nas oclusões, houve uma relação estatisticamente significativa (p = 0,004) entre todos os grupos. Conclusão: O programa de prevenção e promoção de saúde bucal pública foi efetivo na prevenção de cáries, gengivite e má oclusão em crianças com menos de cinco anos de idade.

Efficacy of a public promotion program on children's oral health.

OBJECTIVE: To assess the efficacy of the Baby's Mouth early dental care prevention and promotion program in preventing oral diseases (caries, gingivitis, or malocclusions) in children attended since 2010. METHODS: This was a cross-sectional and cohort study that assessed 252 children between 36 and 60 months of age in both sexes. The children were divided into three groups: G1: effective participants of the program from birth; G2: children who have stopped participating for more than 24 months, and G3: children who have never attended a prevention program. The evaluation was carried out in two stages: first, an interview with the mothers and, afterwards, a clinical children examination to assess the presence of caries, gingivitis, and malocclusion. The chi-squared test was used for statistical analysis between groups (p<0.05). RESULTS: The diseases assessed were: caries (G1: 5.9%, G2: 54.7%, G3: 70%), gingivitis (G1: 8.3%, G2: 17.9%, G3: 40.5%), and malocclusion (G1: 22.6%; G2: 28.6%; G3: 50%). For gingivitis, there was no significant difference when comparing G1 and G2 (p=0.107), but it was significant between G1 and G3 (p<0.001). Regarding malocclusion, a statistically significant relationship was observed (p=0.004) among all groups. CONCLUSION: The prevention and promotion program in public oral health was effective in preventing caries disease, gingivitis, and malocclusion in children under 5 years of age.

Range expansion underlies historical introgressive hybridization in the Iberian hare.

Introgressive hybridization is an important and widespread evolutionary process, but the relative roles of neutral demography and natural selection in promoting massive introgression are difficult to assess and an important matter of debate. Hares from the Iberian Peninsula provide an appropriate system to study this question. In its northern range, the Iberian hare, Lepus granatensis, shows a northwards gradient of increasing mitochondrial DNA (mtDNA) introgression from the arctic/boreal L. timidus, which it presumably replaced after the last glacial maximum. Here, we asked whether a south-north expansion wave of L. granatensis into L. timidus territory could underlie mtDNA introgression, and whether nuclear genes interacting with mitochondria ("mitonuc" genes) were affected. We extended previous RNA-sequencing and produced a comprehensive annotated transcriptome assembly for L. granatensis. We then genotyped 100 discovered nuclear SNPs in 317 specimens spanning the species range. The distribution of allele frequencies across populations suggests a northwards range expansion, particularly in the region of mtDNA introgression. We found no correlation between variants at 39 mitonuc genes and mtDNA introgression frequency. Whether the nuclear and mitochondrial genomes coevolved will need a thorough investigation of the hundreds of mitonuc genes, but range expansion and species replacement likely promoted massive mtDNA introgression.

Dental Anxiety and Behavior in Young Children Undergoing Different Distraction Techniques

Objective: To evaluate anxiety and behavior in groups of children undergoing various distraction techniques during dental treatment in a public clinic. Material and Methods: The research was a randomized study with a systematic convenience sample consisting of 62 children with 4-6 years (5.18±0.77) in both genders; they were divided in four groups (G1 - control group and three experimental Groups: G2 - mirror and conversation, G3 - toys and G4 - children's stories) and evaluated in the first 2 visits to the dentist. Age and previous experience were also evaluated. The Facial Image Scale (FIS) and the Behavior Rating Scale (BRS) were applied, the data was analyzed using the Chi-square test with a significance level of p < 0.05 and the Spearman correlation coefficient. Results: In comparison to the studied variables (anxiety and behavior), the distraction technique during dental care could not reduce anxiety and improve the behavior in all groups in the first visit, but the group receiving the distraction technique with a hand mirror reached the best results in behavior in the second visit (p=0.022; Raj:-2.68). There is no influence on anxiety among children with or without previous experience (p = 0.603), but the age of 4 years showed higher levels of anxiety (p=0.039). Conclusion: Only the distraction technique with the mirror was able to reduce anxiety and improve behavior in the second visit.

The elusive nature of adaptive mitochondrial DNA evolution of an arctic lineage prone to frequent introgression.

Mitochondria play a fundamental role in cellular metabolism, being responsible for most of the energy production of the cell in the oxidative phosphorylation (OXPHOS) pathway. Mitochondrial DNA (mtDNA) encodes for key components of this process, but its direct role in adaptation remains far from understood. Hares (Lepus spp.) are privileged models to study the impact of natural selection on mitogenomic evolution because 1) species are adapted to contrasting environments, including arctic, with different metabolic pressures, and 2) mtDNA introgression from arctic into temperate species is widespread. Here, we analyzed the sequences of 11 complete mitogenomes (ten newly obtained) of hares of temperate and arctic origins (including two of arctic origin introgressed into temperate species). The analysis of patterns of codon substitutions along the reconstructed phylogeny showed evidence for positive selection in several codons in genes of the OXPHOS complexes, most notably affecting the arctic lineage. However, using theoretical models, no predictable effect of these differences was found on the structure and physicochemical properties of the encoded proteins, suggesting that the focus of selection may lie on complex interactions with nuclear encoded peptides. Also, a cloverleaf structure was detected in the control region only from the arctic mtDNA lineage, which may influence mtDNA replication and transcription. These results suggest that adaptation impacted the evolution of hare mtDNA and may have influenced the occurrence and consequences of the many reported cases of massive mtDNA introgression. However, the origin of adaptation remains elusive.