Analysis of Copy Number Variants identifies new candidate genes for Autism Spectrum Disorder and Intellectual Disability

Autism spectrum disorder (ASD) and Intellectual Disability (ID) are complex neuropsychiatric disorders characterized by extensive clinical and genetic heterogeneity and with overlapping risk factors. The aim of my project was to further investigate the role of Copy Numbers Variants (CNVs), identified through genome-wide studies performed by the Autism Geome Project (AGP) and the CHERISH consortium in large cohorts of ASD and ID cases, respectively. Specifically, I focused on four rare genic CNVs, selected on the basis of their impact on interesting ASD/ID candidate genes: a) a compound heterozygous deletion involving CTNNA3, predicted to cause the lack of functional protein; b) a 15q13.3 duplication containing CHRNA7; c) a 2q31.1 microdeletion encompassing KLHL23, SSB and METTL5; d) Lastly, I investigated the putative imprinting regulation of the CADPS2 gene, disrupted by a maternal deletion in two siblings with ASD and ID. This study provides further evidence for the role of CTNNA3, CHRNA7, KLHL23 and CADPS2 as ASD and/or ID susceptibility genes, and highlights that rare genetic variation contributes to disease risk in different ways: some rare mutations, such as those impacting CTNNA3, act in a recessive mode of inheritance, while other CNVs, such as those occurring in the 15q13.3 region, are implicated in multiple developmental and/or neurological disorders possibly interacting with other susceptibility variants elsewhere in the genome. On the other hand, the discovery of a tissue-specific monoallelic expression for the CADPS2 gene, implicates the involvement of epigenetic regulatory mechanisms as risk factors conferring susceptibility to ASD/ID.

The biological basis of autism spectrum disorders: evaluation of oxidative stress and erytrocyte membrane alterations

This case-control study involved a total of 29 autistic children (Au) aged 6 to 12 years, and 28 gender and age-matched typically developing children (TD). We evaluated a high number of peripheral oxidative stress parameters, erythrocyte and lymphocyte membrane functional features and membrane lipid composition of erythrocyte. Erythrocyte TBARS, Peroxiredoxin II, Protein Carbonyl Groups and urinary HEL and isoprostane levels were elevated in AU (confirming an imbalance of the redox status of Au); other oxidative stress markers or associated parameters (urinary 8-oxo-dG, plasma Total antioxidant capacity and plasma carbonyl groups, erythrocyte SOD and catalase activities) were unchanged, whilst peroxiredoxin I showed a trend of elevated levels in red blood cells of Au children. A very significant reduction of both erythrocyte and lymphocyte Na+, K+-ATPase activity (NKA), a reduction of erythrocyte membrane fluidity, a reduction of phospatydyl serine exposition on erythrocyte membranes, an alteration in erythrocyte fatty acid membrane profile (increase in MUFA and in ω6/ω3 ratio due to decrease in EPA and DHA) and a reduction of cholesterol content of erythrocyte membrane were found in Au compared to TD, without change in erythrocyte membrane sialic acid content and in lymphocyte membrane fluidity. Some Au clinical features appear to be correlated with these findings; in particular, hyperactivity score appears to be related with some parameters of the lipidomic profile and membrane fluidity, and ADOS and CARS score are inversely related to peroxiredoxin II levels. Oxidative stress and erythrocyte structural and functional alterations may play a role in the pathogenesis of Autism Spectrum Disorders and could be potentially utilized as peripheral biomarkers.

Non Invasive Tools for Early Detection of Autism Spectrum Disorders

Autism Spectrum Disorders (ASDs) describe a set of neurodevelopmental disorders. ASD represents a significant public health problem. Currently, ASDs are not diagnosed before the 2nd year of life but an early identification of ASDs would be crucial as interventions are much more effective than specific therapies starting in later childhood. To this aim, cheap an contact-less automatic approaches recently aroused great clinical interest. Among them, the cry and the movements of the newborn, both involving the central nervous system, are proposed as possible indicators of neurological disorders. This PhD work is a first step towards solving this challenging problem. An integrated system is presented enabling the recording of audio (crying) and video (movements) data of the newborn, their automatic analysis with innovative techniques for the extraction of clinically relevant parameters and their classification with data mining techniques. New robust algorithms were developed for the selection of the voiced parts of the cry signal, the estimation of acoustic parameters based on the wavelet transform and the analysis of the infant’s general movements (GMs) through a new body model for segmentation and 2D reconstruction. In addition to a thorough literature review this thesis presents the state of the art on these topics that shows that no studies exist concerning normative ranges for newborn infant cry in the first 6 months of life nor the correlation between cry and movements. Through the new automatic methods a population of control infants (“low-risk”, LR) was compared to a group of “high-risk” (HR) infants, i.e. siblings of children already diagnosed with ASD. A subset of LR infants clinically diagnosed as newborns with Typical Development (TD) and one affected by ASD were compared. The results show that the selected acoustic parameters allow good differentiation between the two groups. This result provides new perspectives both diagnostic and therapeutic.

Analysis of the spectrum of the spin-1 biquadratic antiferromagnetic chain

In questo elaborato vengono discusse le catene di spin-1, modelli quantistici definiti su un reticolo unidimensionale con interazione tra siti primi vicini. Fra la ricca varietà di tipologie esistenti è stato scelto di porre attenzione primariamente sul modello antiferromagnetico con interazione puramente biquadratica. Vengono presentati diversi metodi di classificazione degli autostati di tale modello, a partire dalle simmetrie che ne caratterizzano l’Hamiltoniana. La corrispondenza con altri modelli noti, quali il modello XXZ di spin 1/2, la catena di Heisenberg SU (3) ed i modelli di Potts, è utile ad individuare strutture simmetriche nascoste nel formalismo di spin-1, le quali consentono di ricavare informazioni sullo spettro energetico. Infine, vengono presentati risultati numerici accompagnati da alcune considerazioni sulle modifiche dello spettro quando si aggiunge un termine bilineare alla Hamiltoniana biquadratica.

Analysis of aircraft load spectrum by means of flight simulator

Damage tolerance analysis is a quite new methodology based on prescribed inspections. The load spectra used to derive results of these analysis strongly influence the final defined inspections programs that for this reason must be as much as possible representative of load acting on the considered structural component and at the same time, obtained reducing both cost and time. The principal purpose of our work is in improving the actual condition developing a complete numerical Damage Tolerance analysis, able to prescribe inspection programs on typical aircraft critical components, respecting DT regulations, starting from much more specific load spectrum then those actually used today. In particular, these more specific load spectrum to design against fatigue have been obtained through an appositively derived flight simulator developed in a Matlab/Simulink environment. This dynamic model has been designed so that it can be used to simulate typical missions performing manually (joystick inputs) or completely automatic (reference trajectory need to be provided) flights. Once these flights have been simulated, model’s outputs are used to generate load spectrum that are then processed to get information (peaks, valleys) to perform statistical and/or comparison consideration with other load spectrum. However, also much more useful information (loads amplitude) have been extracted from these generated load spectrum to perform the previously mentioned predictions (Rainflow counting method is applied for this purpose). The entire developed methodology works in a complete automatic way, so that, once some specified input parameters have been introduced and different typical flights have been simulated both, manually or automatically, it is able to relate the effects of these simulated flights with the reduction of residual strength of the considered component.

Investigation of spectral stability of X-ray tubes by simulations and experimental spectrum measurements

L'obiettivo di questo lavoro è quello di analizzare la stabilità di uno spettro raggi X emesso da un tubo usurato per analisi cardiovascolari, in modo da verificare il suo comportamento. Successivamente questo tipo di analisi sarà effettuata su tubi CT. Per raggiungere questo scopo è stato assemblato un particolare set-up con un rivelatore al germanio criogenico in modo da avere la miglior risoluzione energetica possibile ed alcuni particolari collimatori così da ridurre il flusso fotonico per evitare effetti di pile-up. Il set-up è stato costruito in modo da avere il miglior allineamento possibile nel modo più veloce possibile, e con l'obiettivo di rendere l'intero sistema portabile. Il tubo usato è un SRM Philips tube per analisi cardiovascolari; questa scelta è stata fatta in modo da ridurre al minimo i fattori esterni (ottica elettromagnetica, emettitori) e concentrare l'attenzione solo sugli effetti, causati dalle varie esposizioni, sull'anodo (roughness e bending) e sul comportamento di essi durante il surriscaldamento e successivo raffreddamento del tubo. I risultati mostrano come durante un'esposizione alcuni fattori di usura del tubo possono influire in maniera sostanziale sullo spettro ottenuto e quindi alterare il risultato. Successivamente, nell'elaborato, mediante il software Philips di ricostruzione e simulazione dello spettro si è cercato di riprodurre, variando alcuni parametri, la differenza riscontrata sperimentalmente in modo da poter simulare l'instabilità e correggere i fattori che la causano. I risultati sono interessanti non solo per questo esperimento ma anche in ottica futura, per lo sviluppo di applicazioni come la spectral CT. Il passo successivo sarà quello di spostare l'attenzione su un CT tube e verificare se l'instabilità riscontrata in questo lavoro è persiste anche in una analisi più complessa come quella CT.

Long-term outcome of symptomatic severe ostial vertebral artery stenosis (OVAS)

The optimal management of patients with symptomatic severe ostial vertebral artery stenosis (OVAS) is currently unclear. We analyzed the long-term outcome of consecutive patients with OVAS who received either medical treatment (MT) or vertebral artery stenting (VAS).

Differences in Demographic Characteristics and Risk Factors in Patients With Spontaneous Vertebral Artery Dissections With and Without Ischemic Events

Spontaneous vertebral artery dissection (sVADs) mainly cause cerebral ischemia, with or without associated local symptoms and signs (headache, neck pain, or cervical radiculopathy), or with local symptoms and signs only.

Single fluoroscopic image based 2D/3D reconstruction of a scaled, patient-specific vertebral model

Purpose Accurate three-dimensional (3D) models of lumbar vertebrae can enable image-based 3D kinematic analysis. The common approach to derive 3D models is by direct segmentation of CT or MRI datasets. However, these have the disadvantages that they are expensive, timeconsuming and/or induce high-radiation doses to the patient. In this study, we present a technique to automatically reconstruct a scaled 3D lumbar vertebral model from a single two-dimensional (2D) lateral fluoroscopic image. Methods Our technique is based on a hybrid 2D/3D deformable registration strategy combining a landmark-to-ray registration with a statistical shape model-based 2D/3D reconstruction scheme. Fig. 1 shows different stages of the reconstruction process. Four cadaveric lumbar spine segments (total twelve lumbar vertebrae) were used to validate the technique. To evaluate the reconstruction accuracy, the surface models reconstructed from the lateral fluoroscopic images were compared to the associated ground truth data derived from a 3D CT-scan reconstruction technique. For each case, a surface-based matching was first used to recover the scale and the rigid transformation between the reconstructed surface model Results Our technique could successfully reconstruct 3D surface models of all twelve vertebrae. After recovering the scale and the rigid transformation between the reconstructed surface models and the ground truth models, the average error of the 2D/3D surface model reconstruction over the twelve lumbar vertebrae was found to be 1.0 mm. The errors of reconstructing surface models of all twelve vertebrae are shown in Fig. 2. It was found that the mean errors of the reconstructed surface models in comparison to their associated ground truths after iterative scaled rigid registrations ranged from 0.7 mm to 1.3 mm and the rootmean squared (RMS) errors ranged from 1.0 mm to 1.7 mm. The average mean reconstruction error was found to be 1.0 mm. Conclusion An accurate, scaled 3D reconstruction of the lumbar vertebra can be obtained from a single lateral fluoroscopic image using a statistical shape model based 2D/3D reconstruction technique. Future work will focus on applying the reconstructed model for 3D kinematic analysis of lumbar vertebrae, an extension of our previously-reported imagebased kinematic analysis. The developed method also has potential applications in surgical planning and navigation.

Vertebral body stenting: a new method for vertebral augmentation versus kyphoplasty

Vertebroplasty and kyphoplasty are well-established minimally invasive treatment options for compression fractures of osteoporotic vertebral bodies. Possible procedural disadvantages, however, include incomplete fracture reduction or a significant loss of reduction after balloon tamp deflation, prior to cement injection. A new procedure called "vertebral body stenting" (VBS) was tested in vitro and compared to kyphoplasty. VBS uses a specially designed catheter-mounted stent which can be implanted and expanded inside the vertebral body. As much as 24 fresh frozen human cadaveric vertebral bodies (T11-L5) were utilized. After creating typical compression fractures, the vertebral bodies were reduced by kyphoplasty (n = 12) or by VBS (n = 12) and then stabilized with PMMA bone cement. Each step of the procedure was performed under fluoroscopic control and analysed quantitatively. Finally, static and dynamic biomechanical tests were performed. A complete initial reduction of the fractured vertebral body height was achieved by both systems. There was a significant loss of reduction after balloon deflation in kyphoplasty compared to VBS, and a significant total height gain by VBS (mean +/- SD in %, p < 0.05, demonstrated by: anterior height loss after deflation in relation to preoperative height [kyphoplasty: 11.7 +/- 6.2; VBS: 3.7 +/- 3.8], and total anterior height gain [kyphoplasty: 8.0 +/- 9.4; VBS: 13.3 +/- 7.6]). Biomechanical tests showed no significant stiffness and failure load differences between systems. VBS is an innovative technique which allows for the possibly complete reduction of vertebral compression fractures and helps maintain the restored height by means of a stent. The height loss after balloon deflation is significantly decreased by using VBS compared to kyphoplasty, thus offering a new promising option for vertebral augmentation.

Desmosomal gene analysis in arrhythmogenic right ventricular dysplasia/cardiomyopathy: spectrum of mutations and clinical impact in practice

Five desmosomal genes have been recently implicated in arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) but the clinical impact of genetics remains poorly understood. We wanted to address the potential impact of genotyping.

Assertive community treatment as part of integrated care versus standard care: a 12-month trial in patients with first- and multiple-episode schizophrenia spectrum disorders treated with quetiapine immediate release (ACCESS trial)

The ACCESS trial examined the 12-month effectiveness of continuous therapeutic assertive community treatment (ACT) as part of integrated care compared to standard care in a catchment area comparison design in patients with schizophrenia spectrum disorders treated with quetiapine immediate release.