People with Autism Spectrum Disorder (ASD) have difficulties in dealing with social interaction (DSM V, 2013). From long time, it has been reported deficit in recognizing gaze, face and intentions of others. Besides that, in the last decade numerous finds have showed an impairment in coding the human body movements. During my PhD I have explored the ability in recognizing the emotional valence of human movements in people with ASD and with typical development. In order to understand developmental changes, I have initially compared this ability between children and adults. In order to understand the role of intelligence, I have included people with high and low functioning ASD, either in children and adults samples. In the first study, we compared TD children and children with ASD with different level of functioning to investigate i) whether the difficulties in ASD population were associated to emotion comprehension or related to a more basic visual processing of biological motion, and ii) to explore whether this ability improves according to the age and to the non-verbal IQ. To this purposes, we presented point-light (PLDs) and full-light (FLDs) version of human movements with three different emotional valence (Happy, Fearful, Neutral). Our main findings showed that: -TD children were more accurate in recognizing the emotional content of body movements when the shape of the body was visible (i.e. FLDs), compared to the view of pure motion information (i.e. PLDs), but there were no differences in RT across display conditions. This result suggests that school-aged children are able to correctly identify the emotional valence of dynamic bodily expression, also when the form information is minimized by presenting PLDs. -According to the emotional content, we found that the valence had an effect on the performance of TD children. In particular, TD children recognized happy bodily expressions with lower accuracy than fearful and neutral movements. Besides, fearful movements were identified more rapidly. We hypothesized that, from an evolutionary point of view, it is more important to recognize fearful signals when they are far from us. Body movements are particularly relevant to convey other people’s state and intentions from a distance, hence bodies are preferential channels to convey fearful signals. -Comparing typical and atypical children, we found that TD were more accurate and more rapid than children with ASD in recognizing the emotional valence of body movements. Moreover, children with HF ASD outperformed children with LF ASD and their performance was significantly predicted from age and nonverbal IQ. This results suggest that the nonverbal abilities have a role in body movement comprehension, probably underpinning the development of compensatory mechanisms which improve with age. -Compared to TD children, children with ASD were impaired not only in understanding the emotional expressions, but also in recognizing the neutral actions. Irrespectively to their IQ level, children with ASD did not benefit from the richness of visual information, and their ability to recognize BM did not change according to the emotional valence. This finding suggests that the difficulty encountered by individuals with ASD in social interaction could be more generally related to biological motion elaboration, rather than being specific for the emotions comprehension. Hence, the lack in emotion recognition in ASD seems to be attributable to a deficit in elaborating motion cues rather than emotional information. To interpret the body movements, children with ASD may adopt compensatory mechanisms, which acquisition seems to be mediated by the nonverbal IQ and improving with age. If this was the case, we should expect an improvement in the ability of recognizing the BM in adults with ASD, with respect to children with ASD, and this improvement should have been correlated with the non-verbal IQ level. To explore this hypothesis, we run a second experiment, were we asked to a group of TD adults, a group of adults with high functioning Autism matched for age and non-verbal IQ, and a group of adults with low functioning Autism matched for age, to performed the same BM recognition task that children did in our first experiment. Results with adults confirmed what we found in children: the emotional valence and the richness of visual information did not modulate the Accuracy and the velocity in recognizing the BM stimuli in ASD, suggesting that the whole body movement is elaborated differently in ASD than in TD adults and that the impairment in ASD is more related to the processing of body movement, rather than being specific for the emotion comprehension. In a third study, we explored the developmental changes in the ability of recognizing the emotional meaning of human whole-body movements from childhood to adulthood. To this aim, we compared the performance between children and adults in each group of Functioning. Results showed that TD adults were more accurate and faster than TD children; FLDs were recognized with higher accuracy and quicker than PLDs; and fearful and neutral movements were recognized significantly better than happy ones. In participants with ASD, adults were significantly more accurate than children, but not faster. Also, we did not find any difference between displays or emotional categories. This results suggest that the understanding of body movements in individuals with ASD improve with age but the mechanism underpinning this ability works differently. In a fourth experiment we investigate whether the impairment in understanding social relevant signals in individuals with high functioning autism (HF ASD) was restricted to body movements or widespread to other social cues, such as facial expressions. Also, we investigated the role of movement in perceiving bodily expressions. To this aim, we explored whether there was a difference in recognizing the body expressions represented by dynamic or static stimuli. Finally, we investigated whether the vision of the body form, compared to the sight of pure motion information, could influence the identification of the emotional content of body movements. We did not found any group differences in accuracy, but there was a group difference in RTs specific for dynamic stimuli (TD were significantly faster than ASD in recognizing PLDs, and marginally in FLDs). These results suggest that HF ASD adopt compensatory mechanism to understand the meaning of facial and bodily expression that allow them to correctly recognize the emotional valence conveyed by face and body movements. However, this mechanism has a cost in terms of rapidity. In particular, ASD seems to need more time to identify dynamic body stimuli but not images of body expressions, suggesting a deficit in processing the actual body motion. In particular, the comprehension of happy body movements result difficult and time consuming. Indeed, happiness was better recognized from facial expressions and was harder to be identified when expressed by body movements. In the fifth experiment we used TMS-adaptation paradigm to explore the existence of a neural system specialized for the elaboration of emotional body movements. We first behaviourally investigated the existence of an adaptation for emotional PLDs. TD adult participant were adapted with point-light video clips depicting fearful, happy or neutral actions and then asked to recognize point-light with same/different emotional content. Results showed an adaptation after-effect only for incongruent stimuli, suggesting the existence of a neural mechanism for perceiving the body emotion specifically. Subsequently, in a TMS experiment we explore the possible brain location of this mechanism. The sites we stimulated are nodes of the neural network responsible for the human motion understanding, and they are reported to be abnormally activated in ASD. We found a reversed after-effect following TMS over aIPS, while the adaptation was still present after stimulation over pSTS and the control site. These results demonstrate that aIPS contains neurons that specifically code for the emotional body expressions, suggesting that the difficulties encountered by individuals with ASD in understanding the emotional signal during social interaction might rely to deficit in this mirror area.

Autism Spectrum Disorder (ASD) is characterized by difficulties in social communication and restricted, repetitive patterns of behavior and interests (American Psychiatric Association, 2013). These key characteristics of autism each create a number of difficulties for individuals with autism and also their parents. It is well established that parenting a child with ASD is stressful, impacting on mental health and overall quality of life (Baker-Ericzn, Brookman-Frazee, & Stahmer, 2005; Davis & Carter, 2008; Eisenhower, Baker, & Blacher, 2005; McStay, Dissanayake, Scheeren, Koot, & Begeer, 2013). However, there are some doubts or gaps in the literature of parenting children with ASD which we addressed in this thesis. Two main goals of this thesis were: a) examining well-being of parents of children with ASD, b) focusing on the parents’ responses to cries of infants with ASD and typical development, considering two types of responses: emotional and physiological. Therefore, this thesis is composed of two parts. The first part in which we aimed to examine well-being of parents of children with ASD has been divided into two sections. The first one dealt with similarities and differences between mothers and fathers in terms of parenting stress, parental mental health and attitude (Chapter 2). Results indicated that mothers of children with ASD reported higher level of depression than fathers and considerable percentages of both mothers and fathers had scores above the clinical cut-points of stress showing that they experience clinically significant levels of parenting stress. Moreover, in terms of parental attitude, mothers engaged with their children in more social exchange than fathers do. 8 The final chapter in Part 1 (Chapter 3) evaluated whether maternal well-being and sense of competence are affected by the outcomes of children receiving intervention. The results suggested that child and family factors, including mothers’ age, were linked to maternal well-being. However treatment-related changes in children’s communication, and parenting satisfaction contributed to well-being above and beyond other factors. A mediation analysis indicated that mothers whose children make treatment gains in communication skills experience a reduction in their level of negative well-being as a consequence of increased parental sense of competence with regards to parenting satisfaction. The second part of the thesis aimed to evaluate emotional and physiological responses of parents of children with ASD during infant crying. This part begins by laying out the theoretical dimensions of the research, and then proceeds as follows: a) the first experiment was concerned with the preparation and validation the cry episodes as unpleasant acoustic stimuli (Chapter 4); b) the second one examined emotional and physiological responses of non-parents adults while listening of infant crying (Chapter 5); c) the last experiment focused on parents of children with ASD and parents of typically developing children, examining their emotional and physiological responses during the listening of crying of children with ASD and of typically developing (TD) children (Chapter 6). Findings from Chapter 4 indicated that all cry episodes, regardless of the types (e.g., cries of children with ASD or cries of typically developing children) were perceived as unpleasant by non-parents adults. The following study in Chapter 5 showed that cries of children with ASD (ASD cry) were reported more stressed, aroused and less pleasant compare to cries of typically developing children (TD cry) by non-parents adults; however, similar pattern was not seen in the physiological responses of listeners. Results 9 from the final experiment (Chapter 6) showed that parents of children with ASD and parents of TD children were not differentiated in their self-reports of stress, arousal and valence levels for ASD cry and TD cry. However, their physiological responses showed that parents of children with ASD have higher heart rate than parents of TD children during ASD cry and TD cry. Moreover, the analysis on the comparisons between ASD cry and TD cry suggested both parents perceived ASD cry more stressful, aroused and less pleasant, but physiological responses of parents of children with ASD did not show the differences between ASD cry and TD cry. The overall structure of the thesis takes the form of seven chapters, including a general introduction (Chapter 1). The importance of those findings for future theoretical and clinical work is considered in detail in the end of related chapters and in the general discussion chapter (Chapter 7).

In the last two decades several studies have been trying to explore a possible role for gut microbiota in Autism Spectrum Disorders (ASD), supported by the high incidence of gastrointestinal disorders among ASD children and by the now well recognized existence of the brain-gut-microbiota axis (the complex system of bidirectional interactions between central nervous system, gastrointestinal tract and microorganisms inhabiting the gut). Nevertheless, results about alterations in gut microbiota composition and/or activity in ASD are to date strongly contrasting. A possible explanation could be that these studies tend to treat ASD as a unique pathology, whereas it includes different cognitive-behavioural phenotypes. Moreover, they do not consider factors which are important for children’s gut flora development, such as type of delivery, nutritional history (e.g. formula milk during lactation) and medical history (e.g. antibiotics intake) as well as factors that may affect the present composition of microbiota, such as the current diet (e.g. the strong food selectivity that often occurs in ASD children) and the presence of gastrointestinal disorders. In this study, I developed an interview to parents to assess whether there are differences related to the above mentioned aspects between ASD children and typically developing children and among ASD themselves, considering differences in cognitive level and severity of autistic traits. I also explored the use of special diets such as gluten-, lactose and casein free diets, the reasons for their adoption and the possible benefits for the child. Moreover, I decided to include in this interview also a section dedicated to parental difficulties in managing mealtime in order to collect information useful to plan future interventions. I found differences between ASD- and typical children in the incidence of gastrointestinal disorders and food selectivity. Especially, some children initially eat everything and then switch to a more and more restricted diet. This could be considered an early symptom of the pathology. I also found an association between gastrointestinal disorders and severity of autistic traits. Furthermore, I collected faecal samples from ASD families (two parents, an ASD child and a typically developing sibling) and analysed them with metaproteomics and bioinformatics techniques in order to assess microbiota activity and evaluate it in light of ASD phenotype, nutritional habits, gastrointestinal disorders and genetic proximity. Demonstrating the existence of a different microbiota composition in ASD or at least in a subgroup could allow to identify a biomarker of a possible development of ASD and to design preventive interventions, even through probiotics intake. Moreover, it could help to better understand the molecular mechanism underlying this pathology.

Clinical and research practice in the context of Autism rapidly evolved in the last decades. Finer diagnostic procedures, evidence-based models of intervention and higher social inclusivity significantly improved the possibility for autistic children to participate in the fabric of social life. In terms of health best practices, gold-standard procedures still need to be improved, and bridging research and clinical practice still presents several challenges. From the clinical standpoint, the role of process variables, predictors, mechanisms, and timing of change still requires extensive investigation in order to explain response variability and design optimized interventions, tailored to individual needs and maximally effective. Observational techniques represent the elective research methods in child development, especially in clinical contexts, due to their non-invasiveness. However, they still suffer from limited objectivity and poor quantification. Further, their main disadvantage is that they are highly time-consuming and labor-intensive. The aim of this thesis was moving forward to promote translational research in clinical practice of Autism intervention with preschool children. At first, we tried to design and apply quantitative observational techniques to longitudinally study treatment response trajectories during developmental intervention. We tried to characterize different response profiles, and which baseline predictors were able to predict the response over time. Secondly, we investigated mechanisms of change. In particular, we focused on the role of the child-therapist interaction dynamics as a possible active mediator of the process of intervention, especially in the developmental framework that stresses the importance of interpersonal aspects. We also aimed at understanding whether certain time-windows during the intervention were particularly predictive of the response, as well as which specific interaction aspects played a role. Finally, to promote the translational application of observational methods and to improve objective quantification, we proposed and validated an Artificial Intelligence (AI) system to automate data annotation in unconstrained clinical contexts, remaining completely non-invasive and dealing with the specific noisy data that characterize them, for the analysis of the child-therapist acoustic interaction. This effort represents a base building block enabling to employ downstream computational techniques greatly reducing the need for human annotation that usually prevents the application of observational research to large amounts of data . We discuss our findings stressing the importance of assuming a developmental framework in Autism, the key role of the interpersonal experience also in the clinical context, the importance of focusing on trajectories of change and the important need to promote the acquisition of large amounts of quantitative data from the clinical contexts exploiting AI-based systems to assist clinicians, improving objectivity, enabling treatment monitoring, and producing precious data-driven knowledge on treatment efficacy.

Mother child relationship is the first and the most important social relationship as it has implications on psychological and neural development of the individual. Here we investigated mother child relationship focusing on different aspects and using a combination of behavioural and neuroimaging techniques. In the first study we addressed the association between brain connectivity and interpersonal competences which are at the basis of every social interaction including the ones involved in mother-child relationship. Several studies suggests that higher White Matter (WM) integrity - an index of increased brain connectivity - , is associated with better cognition and behavioural performance. To test the hypothesis that higher WM integrity is associated with higher interpersonal competence we used Diffusion Tensor Imaging (DTI), a neuroimaging technique which allows to study in vivo the anatomy of boundless of axons conveying information in the brain. Then we correlated this information with a self-reported measure of interpersonal competences: the Adolescent Interpersonal Competence Questionnaire (AICQ). Results indicate that Interpersonal competence is associate with higher WM integrity in several major tracts of the right hemisphere, in specific the uncinate fasciculus, the cingulum, the forceps minor, the infero-fronto occipital fasciculus, the inferior longitudinal fasciculus, and the superior longitudinal fasciculus. These results provide the first direct analysis of the neuroanatomical basis of interpersonal competencies and young adult self-reported skills in social contexts. In the second work we used the same paradigm to test one of the main assumption of the attachment theory which states that social skills highly depends on the quality of attachment relationship. Results show higher integrity in four white matter association fibers in the left hemisphere: Uncinate Fasciculus, Cingulum, Superior Longitudinal Fasciculus and Inferior Fronto Occipital Fasciculus. This result supports the idea that the quality of the attachment relationship influences the emotional and social life of the individual from childhood to adulthood. Furthermore, the research represents an explorative approach to the study of mother-child relationship in healthy population, demonstrating the feasibility of using neuroimaging tools coupled with clinical investigations. Together those studies show that efficient structural connectivity is linked with secure attachment, improved social cognition and cognitive ability. Similarities and differences emerged in these studies will be discussed at the end of Chapter 3 in particular regarding left and right hemisphere specialization. In the second part of the thesis we switched the focus on parenting behaviour. Evidence from the literature suggest an association between Axonal Integrity measured with FA and functional connectivity measured with TMS in two region involved in preparing ad executing actions: premotor and motor cortex. Moreover neuroimaging reveals that infant cries activate parts of the premotor cortical system. In line with this evidence we linked parenting and brain functional connectivity conducting a study on motor cortex excitability in response of infant cries. We used event-related transcranial magnetic stimulation (TMS) to investigated the presence and the time course of modulation of motor cortex excitability in young adults who listened to infant cries. TMS was delivered from 0 to 250 ms from sound onset in six steps of 50 ms in 10 females and 10 males. Motor Evoked Potentials (MEPs) were recorded from the biceps brachii (BB) and interosseus dorsalis primus (ID1) muscles. Results indicate an excitatory modulation of MEPs at 100 ms from the onset of the infant cry specific to females and to the ID1 muscle. This modulation is considered as automatic response to natural cry as it was not present in response of control sounds and the effect is found at 100-ms latency which make this modulation not compatible with a voluntary reaction to the stimulus but suggests an automatic, bottom-up audiomotor association. These results indicate that the brains of adult females appear to be tuned to respond to infant cries with automatic motor excitation. This effect may reflect the greater and longstanding burden on females in caregiving infants. The second part of the thesis continue with a study addressing the natural condition in which baby cries arise when the parent is not attending for infant stimulation. In this study we investigated how infant crying, compared to control sounds, captures adults’ attentive resources. Participants were all nulliparous women and men, we investigated the effects of different sounds on cerebral activation of the default mode network (DMN) while listeners engaged in two different kind of tasks: one designed to activate the DMN ( self-referential decision task) and one designed to deactivate the DMN (syllabic counting tasks). We found a strong deactivation of DMN in woman during baby cry which suggest a shift of attention from self-referential thinking toward the baby cry stimuli. In men we found instead a weaker deactivation of DMN during woman cry while their attention was directed toward an external task and simultaneously a sudden woman crying arise. Gender differences found in our studies and in the literature will be discussed. In the third part of the thesis we investigated the ability to discriminate synchrony and asynchrony during interaction between mother and child with typical or atypical development. We tested two kind of population: in the first study we compared parents of children with Autism Spectrum Disorder (ASD) to parents of children with Typical Development (TD). In the second study we used the Autistic Quotient questionnaire to divide the sample in two groups according with their autistic traits. The relevance of this task is due to the extreme importance to promptly individuate cues of abnormal social behavior in those cases in which the child might shows deficit in the social development. We hypothesized that individual related with a child with ASD or an individual with high autistic traits, might show similar social difficulties as the individual with ASD finding more challenging to detect cue of appropriate or unappropriated social behavior. To test this hypothesis we asked parents of children with ASD and parents of children with TD to judge video of interactions between mothers and child with ASD and mothers and child with TD. Each video were 20s long and depicted either a synchrony or asynchrony interaction, as categorized by an expert clinician. Contrary to our initial hypothesis results indicate that parents of children with ASD are as accurate as parents of children with TD in discriminating synchrony and asynchrony interaction with ASD, however they are less accurate than parents of children with TD in judging interaction with TD. In the second study by testing individual with higher autistic traits (HAQ group) versus lower autistic traits (LAQ group) we confirmed this trend. Using the same paradigm we found that both groups were less accurate during asynchrony interaction. However HAQ was more accurate in judging synchrony interaction with ASD while LAQ was more accurate in judging synchrony interaction with TD. This result indicate a facilitation effects in understanding interaction which include people that share similar characteristic with the observer disconfirming the hypothesis that people with higher autistic traits would have more difficulties in understanding social interactions and pointing the attention on other factors which might contribute during this process. A discussion on the need of further investigation using neuroimaging techniques to understand similarities and differences on neural processing of social interactions is provided at the end of Chapter 4.

La capacità di esprimere adeguatamente le emozioni attraverso l’uso di strategie adattive di regolazione delle emozioni è cruciale per il sano sviluppo dei bambini. Secondo la teoria dell'attaccamento di Bowlby, la reattività e la regolazione emotiva dei bambini sono correlate ai loro stili di attaccamento. Il presente studio mira ad indagare se l'interazione tra la classificazione di attaccamento e l’espressione emotiva può predire i cambiamenti nella reattività fisiologica dei bambini durante un compito stressante. In un primo momento (T1), le rappresentazioni di attaccamento di N = 59 bambini in età scolare sono state misurate utilizzando la Child Attachment Interview. In un secondo momento (T2), i bambini hanno sostenuto un’impegnativa procedura di laboratorio standardizzata in cui dovevano cercare di risolvere una serie di puzzles risolvibili e irrisolvibili. I livelli di conduttanza cutanea (SCL), l'aritmia sinusale respiratoria (RSA) ed i livelli di cortisolo sono stati registrati come indici fisiologici durante l’intero compito. Le espressioni emotive facciali dei bambini (tristezza e vergogna) sono state codificate durante il quarto puzzle irrisolvibile. I risultati hanno evidenziato che l'espressione dell'emozione modera la relazione tra l'attaccamento evitante e le risposte fisiologiche: in alcuni compiti sono stati mostrati una RSA più bassa e dei SCL più alti; inoltre, l'interazione tra attaccamento ed espressione emotiva di ritiro era associata ad un aumento quasi significativo dei livelli di cortisolo durante l’intera esecuzione dei puzzles. I risultati rivelano che la presenza di attaccamento evitante modulata dall'espressione emotiva di ritiro induce uno stress fisiologico più elevato nei bambini. Comprendere i fattori che influenzano la reattività allo stress nei bambini è essenziale nello sviluppo di interventi che promuovano un adattamento funzionale in risposta al disagio.