Sviluppo di tecniche di analisi ifenate per la quantificazione di biomarcatori di esposizione a tossici ambientali: gli acidi mercapturici

Human biomonitoring (HBM) is an ideal tool for evaluating toxicant exposure in health risk assessment. Chemical substances or their metabolites related to environmental pollutants can be detected as biomarkers of exposure using a wide variety of biological fluids. Individual exposure to aromatic hydrocarbon compounds (benzene, toluene, and o-xylene –“BTX”) were analysed with a liquid chromatography coupled to electrospray ionisation-mass spectrometry (μHPLC-ESI-MS/MS) method for the simultaneous quantitative detection of the BTX exposure biomarker SPMA, SBMA and o-MBMA in human urine. Urinary S-phenylmercapturic acid (SPMA) is a biomarker proposed by the American Conference of Governmental Industrial Hygienists (ACGIH) for assessing occupational exposure to benzene (Biological Exposure Index of 25 microg/g creatinine). Urinary S-benzylmercapturic (SBMA) and o-methyl S-benzyl mercapturic acid (o-MBMA) are specific toluene and o-xylene metabolites of glutathione detoxicant pathways, proposed as reliable biomarkers of exposure. To this aim a pre-treatment of the urine with solid phase extraction (SPE) and an evaporation step were necessary to concentrate the mercapturic acids before instrumental analysis. A liquid chromatography separation was carried out with a reversed phase capillary column (Synergi 4u Max-RP) using a binary gradient composed of an acquous solution of formic acid 0.07% v/v and methanol. The mercapturic acids were determinated by negative-ion-mass spectrometry and the data were corrected using isotope-labelled analogs as internal standards. The analytical method follows U.S. Food and Drug Administration guidance and was applied to assess exposure to BTX in a group of 396 traffic wardens. The association between biomarker results and individual factors, such as age, sex and tobacco smoke were also investigated. The present work also included improvements in the methods used by modifying various chromatographic parameters and experimental procedures. A partial validation was conducted to evaluate LOD, precision, accuracy, recovery as well as matrix effects. Higher sensitivity will be possible in future biological monitoring programmes, allowing evaluation of very low level of BTX human exposure. Keywords: Human biomonitoring, aromatic hydrocarbons, biomarker of exposure, HPLC-MS/MS.

La comunicazione della cooperazione allo sviluppo: gli Enti Pubblici. Analisi discorsive dei Siti Web

Our research takes place in the context of a discipline kwown as Communication for Development, sited inside the field of Communication for Social Change, characterized by the use of interpersonal ad mass communication theories and tools, applyied to international development cooperation. Our study aims at pointing out a change of paradigm in this field: our object is Public Administration’s communication, therefore, what we suggest is a shift from Communication for Development, to Development Communication. The object of our study, hence, becomes the discourse itself, in its double action of representation and construction of reality. In particular, we are interested in the discourse’s tribute to the creation of a collective immagination, wich is the perspective towards which we have oriented the analysis, through a structuralist semoitics-based methodology integrated with a socio-semiotic approach. Taking into consideartion the fact that in our contemporary society (that is to say a ‘Western’ and ‘First World’ society), the internet is a crucial public space for the mediation and the management of collective immagination, we chose the web sites of Public Bodies which are dedicated to International Cooperation has our analysis corpus. This, due to their symbolic and ideologic significance, as well as for the actual political responsibility we think these web sites should have. The result of our analysis allows us to suggest some discoursive strategies used in the web sites of Public Bodies. In these sites, there is a tendency to shift the discourses around international cooperation from the ideological axis - avoiding in so doing to explicit a political statement about the causes of injustices and un-balances which lead to the necessity of a support in development (i.e. avoiding to mention values such as social justice and democracy while acknowledging socio-economical institutions which contribute to foster underdevelopment on a global scale) -, to the ethical axis, hence referring to moral values concerning the private sphere (human solidarity and charity), which is delegated mainly to non governamental associations.

Innovazione organizzativa e istituzionale della pubblica amministrazione. Un approccio interpretativo e l'analisi di un dispositivo partecipativo nel settore della cultura

The thesis aims at inquiring into the issue of innovation and organizational and institutional change in the public administration with regard to the increasingly massive adoption of participatory devices and practices in various arenas of public policies. The field of reference regards transformations of the types of public actions and regulation systems, concerning governance. Together with the crisis of the public function and of the role played by the insitutions what is emerging are different levels of governement, both towards an over national and a local direction, and a plurality of social interlocutors, followed by a post-bureaucratic pattern of the public administration that is opening itself in the direction of environment and citizens. The public adminstration is no longer considered an inert object within the bureaucratic paradigm but as a series of communicative processes, choices, cultures and practices that actively builds itself and the environment it interacts with. Therefore, the output of the public administration isn’t the simple service being supplied but the relationship enacted with the citizen, relationship that becomes the constituent basis of adminstrative processes. The intention of thesis is to take into consideration the relation between innovation of the public administration and participatory experimentations and implementations regarded as exchanges in which citizens and the public administration hold talks and debates. The issue of the organizational change of the public administration as output and effect of inclusive deliberative practices has been analysed starting from an institutionalist approach, in other words examining the constituent features of institutions, “rediscovering” them with regard to their public nature, their ability to elaborate collective values and meanings, the social definition of problems and solutions. The participatory device employed by the Forlì city council that involved enterprises and cultural associations of the area in order to build a participatory Table, has been studied through a qualitative methodology (participant observation and semi-strutctured interviews). The analysis inquired into the public nature both of the participatory device and the administrative action itself as well as into elements pertaining the deliberative setting, the regulative reference framework and the actors which took part in the process.

Analisi critico-comparativa delle traduzioni italiane di Un amour de Swann di Marcel Proust

Affrontare l’analisi critica delle traduzioni italiane di Un amour de Swann di Marcel Proust implica necessariamente un attento esame dello stile esemplare di un autore, che ha modificato, in larga misura, la concezione del romanzo e della scrittura stessa; in secondo luogo significa considerare l’altrettanto complesso lavorio intellettuale rappresentato dalla traduzione, in qualità di atto critico fondamentale. Prenderemo in esame nel capitolo primo la genesi del TP, le sue specificità narratologiche e la ricezione critica francese e italiana, per comprendere appieno l’originalità di Proust rispetto al panorama letterario dell’epoca. Nella seconda parte del primo capitolo delineeremo un excursus lungo la storia di tutte le traduzioni italiane del romanzo, accordando particolare attenzione alla figura di ogni traduttore: Natalia Ginzburg (1946), Bruno Schacherl (1946), Armando Landini (1946), Oreste Del Buono (1965), Giovanni Raboni (1978), Maria Teresa Nessi Somaini (1981), Gianna Tornabuoni (1988), Eurialo De Michelis (1990) e il traduttore, rimasto anonimo, della casa editrice La Spiga Languages (1995). Nel secondo capitolo analizzeremo la peculiarità stilistica più nota dell’autore, la sintassi. I lunghi periodi complicati da un intreccio di subordinate e sciolti solo con il ricorso a vari nessi sintattici contribuiscono a rendere la sintassi proustiana una delle sfide maggiori in sede traduttiva. Nel capitolo successivo accorderemo attenzione all’eteroglossia enunciativa, espressa nei diversi idioletti del romanzo. In sede contrastiva affronteremo la questione della trasposizione dei niveaux de langue, per poter valutare le scelte intraprese dai traduttori in un ambito altamente complesso, a causa della diversità diafasica intrinseca dei due codici. All’interno del medesimo capitolo apriremo una parentesi sulla specificità di una traduzione, quella di Giacomo Debenedetti, effettuata seguendo una personale esegesi dello stile musicale e armonico di Proust. Riserveremo al capitolo quarto lo studio del linguaggio figurato, soffermandoci sull’analisi delle espressioni idiomatiche, che vengono impiegate frequentemente da alcuni personaggi. Rivolgeremo uno sguardo d’insieme alle strategie messe in atto dai traduttori, per cercare un equivalente idiomatico o per ricreare il medesimo impatto nel lettore, qualora vi siano casi di anisomorfismo. Analizzeremo nel quinto capitolo la terminologia della moda, confrontando il lessico impiegato nel TP con le soluzioni adottate nei TA, e aprendo, inevitabilmente, una parentesi sulla terminologia storica del settore. A compimento del lavoro presenteremo la nostra proposta traduttiva di un capitolo tratto dal saggio Proust et le style di Jean Milly, per mettere in luce, tramite la sua parola autorevole, la genialità e la complessità della scrittura proustiana e, conseguentemente, il compito ardimentoso e ammirevole che è stato richiesto ai traduttori italiani.

Analisi spaziale dei dati entomologici in ambiente GIS: prospettive di utilizzo nell'agricoltura di precisione

Scopo della ricerca è stato definire le dinamiche spazio-temporali degli insetti studiati mediante l’impiego di tecniche geostatistiche. La ricerca è stata condotta su due casi studio, il primo riguardante tre specie di Elateridi di elevata importanza economica su scala aziendale, il secondo inerente al monitoraggio di Diabrotica virgifera virgifera (diabrotica del mais) su scala regionale. Gli scopi specifici dei due casi studio sono stati: Caso studio 1 a) Monitorare l’entità della popolazione di Elateridi su scala aziendale mediante approccio geostatistico. b) Elaborazione di mappe di distribuzione spaziale interfacciabili all’ambiente GIS. c) Individuare i fattori predisponenti l’infestazione. d) Verificare la necessità dell’impiego di mezzi chimici per il controllo delle specie dannose. e) Proporre strategie alternative volte alla riduzione dell’impiego di trattamenti geodisinfestanti. Caso studio 2 a) Studiare la distribuzione spaziale su scala regionale la popolazione del fitofago D. virgifera virgifera. b) Applicare a livello regionale una griglia di monitoraggio efficace per studiarne la diffusione. c) Individuare le aree a rischio e studiare i fattori predisponenti l’infestazione e diffusione. d) Ottimizzare ed economizzare il piano di monitoraggio.

Analisi genetico-molecolare dei linfomi a cellule T periferiche

Molecular profiling of Peripheral T-cell lymphomas not otherwise specified Peripheral T-cell lymphomas (PTCLs) are a heterogeneous group of tumors that the WHO classification basically subdivides into specified and not otherwise specified (NOS). In Western countries, they represent around 12% of all non-Hodgkin's lymphomas. In particular, PTCL/NOS is the commonest subtype, corresponding to about 60-70% of all T-cell lymphomas. However, it remains a complex entity showing great variety regarding either morphology, immunophenotype or clinical behavior. Specially, the molecular pathology of these tumors is still poorly known. In fact, many alteration were found, but no single genes were demonstrated to have a pathogenetic role. Recently, gene expression profiling (GEP) allowed the identification of PTCL/NOS-associated molecular signatures, leading to better understanding of their histogenesis, pathogenesis and prognostication. Interestingly, proliferation pathways are commonly altered in PTCLs, being highly proliferative cases characterized by poorer prognosis. In this study, we aimed to investigate the possible role in PTCL/NOS pathogenesis of selected molecules, known to be relevant for proliferation control. In particular, we analyzed the cell cycle regulators PTEN and CDKN1B/p27, the NF-kB pathway, and the tyrosin kinase PDGFR. First, we found that PTEN and p27 seem to be regulated in PTCL/NOS as in normal T-lymphocytes, as to what expression and cellular localization are concerned, and do not present structural abnormalities in the vast majority of PTCL/NOS. Secondly, NF-kB pathway appeared to be variably activated in PTCL/NOS. In particular, according to NF-kB gene expression levels, the tumors could be divided into two clusters (C1 and C2). Specially, C1 corresponded to cases presenting with a global down-regulation of the entire pathway, while C2 showed over-expression of genes involved in TNF signaling. Notably, by immunohistochemistry, we showed that either the canonical or the alternative NK-kB pathway were activated in around 40% of cases. Finally, we found PGDFRA to be consistently over-expressed (at mRNA and protein level) and activated in almost all PTCLs/NOS. Noteworthy, when investigating possible causes for PDGFRA deregulation, we had evidences that PDGFR over-expression is due to the absence of miR-152, which appeared to be responsible for PDGFRA silencing in normal T-cells. Furthermore, we could demonstrate that its aberrant activation is sustained by an autocrine loop. Importantly, this is the first case, to the best of our knowledge, of hematological tumor in which tyrosin kinase aberrant activity is determined by deregulated miRNA expression and autocrine loop activation. Taken together, our results provide novel insight in PTCL/NOS pathogenesis by opening new intriguing scenarios for innovative therapeutic interventions.

Costruzione di un Sistema Informativo Geografico Digitale per analisi ambientali

Questo lavoro ha come obiettivo l’utilizzo del Geographical Information System (GIS) per effettuare analisi di sicurezza, monitoraggio e valutazioni di impatto ambientale. Oggi, la totalità delle operazioni GIS possono essere svolte con software open source e in questa sedi si è scelto di utilizzare il GIS GRASS (Geographic Resources Analysis Support System) disponibile nei termini della GNU public license (GPL), mostrando l’usabilità e le notevoli potenzialità di tale software, nonché la qualità dei prodotti ottenibili, mai inferiori ai prodotti e agli strumenti messi a disposizione dai più radicati e diffusi programmi proprietari. Nel capitolo 4, vedremo l’applicazione all’analisi delle conseguenze di ipotetici incidenti, durante le operazioni di dismissione dell’impianto di processamento del combustibile nucleare, di Bosco Marengo (AL). Nel capitolo 5, vedremo applicazioni nel campo del monitoraggio della qualità dell’aria tramite analisi di immagini satellitari.

Rapporti tra tettonica e clima durante l'esumazione nelle Alpi Centrali. Evidenze dalla termocronologia e dall'analisi strutturale lungo il traforo ferroviario del Sempione

(U-Th)/He and fission-track analyses of apatite along deep-seated tunnels crossing high-relief mountain ranges offer the opportunity to investigate climate and tectonic forcing on the topographic evolution. In this study, the thermochronologic analysis of a large set of samples collected in the Simplon railway tunnel (western-central Alps; Italy and Switzerland) and along its surface trace, coupled with kinematic and structural analysis of major fault zones intersecting the tunnel, constrains the phenomena controlling the topographic and structural evolution, during the latest stage of exhumation of the Simplon Massif, and the timing in which they operated. The study area is located at the western margin of the Lepontine metamorphic dome where a complex nappe-stack pertaining to the Penninic and Ultrahelvetic domains experienced a fast exhumation from the latest Oligocene onward. The exhumation was mainly accommodated by a west-dipping low-angle detachment (the Simplon Fault Zone) which is located just 8 km to the west of the tunnel. However, along the section itself several faults related to two principal phases both with important dip-slip kinematics have been detected. Cooling rates derived from our thermocronological data vary from about 10 °C/Ma at about 10 Ma to about 35 °C/Ma in the last 5 Ma. Such increase in the cooling rate corresponds to the most important climatic change recorded in the northern hemisphere in the last 10 Ma, i.e. the shift to wetter conditions at the end of the Messinian salinity crisis and the inception of glacial cycles in the northern hemisphere. In addition, (U-Th)/He and fission-track age patterns lack of important correlation with the topography suggesting that the present-day relief morphology is the result of recent erosional dynamics. More in details, the (U-Th)/He tunnel ages show an impressive uniformity at 2 Ma, whereas cooling rates calculated at 1 Ma increase towards the two major valleys. This indicates a focusing of erosive processes in the valleys which led to the shaping of present-day topography. Structural analysis documents the presence of two phases of brittle deformation postdating the metamorphic phases in the area. The first one is directly related to the last phase of activity along the Simplon Fault Zone and is characterized by extension towards SO and vertical shortening. The young one is characterized by extension towards NO and horizontal shortening in a along the NE-SO direction. Structures related to the first phase of brittle deformation generate important variations in the older ages' dataset, until 3 Ma, suggesting that tectonics controlled rocks exhumation up to that age. Structures related to the second phase generate some variations also in the younger age dataset, highlighting the activity of faults bordering the massif and suggesting a continuous activity also after 2 Ma. However, most of (U-Th)/He tunnel ages, varying slightly around 2 Ma, document that the Simplon area has experienced primarily erosional exhumation in this time span. In conclusion, all our data suggest that in the central Italian Alps the climatic signal gradually overrode the tectonic effects after about 5 Ma, as a consequence of the climatic instability started at end of Messinian salinity crisis and improved by the onset of glaciations in the northern hemisphere.

Gli assetti dipartimentali ospedalieri tra strutture formali e organizzazione informale

Il confronto in corso tra gli esperti di management sanitario sui dipartimenti ospedalieri, la crescente attenzione sui modelli di organizzazione a rete e le indagini sui servizi sanitari condotte con strumenti di analisi dei network hanno rappresentato la base su cui sviluppare il disegno dello studio. La prospettiva relazionale e le tecniche di social network analysis (Sna) sono state impiegate in un indagine empirica effettuata presso tre Dipartimenti Ospedalieri dell’Azienda USL di Bologna per osservare la struttura delle relazioni che intercorrono nell’ambito dei dipartimenti, tra le unità operative e tra i clinici, al fine di assicurare il quotidiano svolgersi dei processi clinico assistenziali dei pazienti. L’indagine si è posta tre obiettivi. Il primo è quello di confrontare la rete delle relazioni “reali” che intercorrono tra unità operative e tra clinici con le relazioni “progettate” attraverso l’afferenza delle unità operative nei dipartimenti e dei singoli clinici nelle unità operative. In sostanza si tratta di confrontare, con intenti esclusivamente conoscitivi, la struttura organizzativa formale – istituzionale con quella “informale”, che emerge dalle relazioni giornaliere tra i professionisti. In secondo luogo si intende comprendere se e come i fattori di natura attributiva che caratterizzano i singoli rispondenti, (es. età, sesso, laurea, anni di permanenza in azienda, ecc.) incidano sulla natura e sull’intensità delle relazioni e delle collaborazioni intrattenute con i colleghi all’interno dell’azienda. L’analisi ha un intento “esplicativo”, in quanto si cerca di indagare come le similitudini nelle caratteristiche individuali possano o meno incidere sull’intensità degli scambi e quindi delle collaborazioni tra professionisti. Il terzo obiettivo è volto a comprendere se e come i fattori attributivi e/o relazionali siamo in grado di spiegare l’attitudine mostrata dai singoli professionisti rispetto l’adozione di un approccio alla pratica clinica ispirato all’Evidence based medicine. Lo scopo è quello di verificare se la disponibilità / orientamento ad operare in una prospettiva evidence based sia più legata ad elementi e caratteristiche personali piuttosto che all’influenza esercitata da coloro con i quali si entra in contatto per motivi lavorativi. La relativa semplicità della fase di indagine ha indotto ad arricchire i contenuti e gli obiettivi originari del lavoro allo scopo di correlare indicatori relazionali e attributivi con indicatori di “performance”, in particolare di efficienza e appropriatezza. Le relazioni sono state rilevate attraverso un questionario sociometrico inserito in uno spazio web accessibile dalla rete ospedaliera e compilato online da parte dei medici. Il questionario è stato organizzato in tre sezioni: la prima per la raccolta di informazioni anagrafiche e dati attributivi dei clinici; la seconda volta a raccogliere i dati relazionali, funzionali e di consulenza, verso le equipe di professionisti (unità operative) e verso i singoli colleghi clinici; la terza sezione è dedicata alla raccolta di informazioni sull’utilizzo delle evidenze scientifiche a supporto della propria pratica clinica (consultazione di riviste, banche dati, rapporti di HTA, etc,) e sulla effettiva possibilità di accesso a tali strumenti. L’azienda ha fornito i dati di struttura e la base dati degli indicatori di attività delle UO arruolate nello studio. La compliance complessiva per i tre dipartimenti è stata pari a circa il 92% (302 rispondenti su un campione di 329 medici.). Non si sono rilevate differenze significative sulla compliance per i tre dipartimenti considerati. L’elaborazione dei dati è stata effettuata mediante specifici software per l’analisi delle reti sociali, UCINET 6 per il calcolo degli indicatori relazionali (centralità, densità, structural holes etc.), e Pajek per l’analisi grafica dei network. L’ultima fase è stata realizzata con l’ausilio del software statistico STATA vers. 10. L’analisi dei risultati è distinta in due 2 fasi principali. In primis è stato descritto il network di relazioni professionali rilevate, sono stai calcolati i relativi indicatori di centralità relazionale e verificato il grado di sovrapposizione tra struttura formale dei dipartimenti in studio con le relazioni informali che si stabiliscono tra di essi nell’ambito clinico. Successivamente è stato analizzato l’impatto che le relazioni esercitano sulla propensione da parte dei singoli medici a utilizzare nuove evidenze scientifiche I primi risultati emersi dallo studio forniscono interessanti evidenze, con particolare riguardo al dato di un discreto grado di “sovrapposizione” tra struttura formale e informale delle unità organizzative in studio e a correlazioni significative tra fattori relazionali e attitudine dei medici verso l’utilizzo dell’approccio EBM. Altre evidenze, in specie la correlazione tra “centralità” degli attori organizzativi e alcuni indicatori di performance /appropriatezza, meritano ulteriori approfondimenti e una definitiva validazione. In conclusione lo studio dimostra che la prospettiva relazionale e la Sna consentono di porre in evidenza caratteristiche dei dipartimenti, dei suoi attori e delle loro reti di reciproche relazioni, in grado di favorire la comprensione di alcune dinamiche ricercate proprio attraverso l’organizzazione dipartimentale e quindi di specifico interesse per il management, i clinici e quanti altri impegnati nella gestione e nello sviluppo di questo modello di organizzazione dell’ospedale.

Genetica molecolare dell'autismo: studi di associazione ed analisi di geni candidati

Autism is a neurodevelpmental disorder characterized by impaired verbal communication, limited reciprocal social interaction, restricted interests and repetitive behaviours. Twin and family studies indicate a large genetic contribution to ASDs (Autism Spectrum Disorders). During my Ph.D. I have been involved in several projects in which I used different genetic approaches in order to identify susceptibility genes in autism on chromosomes 2, 7 and X: 1)High-density SNP association and CNV analysis of two Autism Susceptibility Loci. The International Molecular Genetic Study of Autism Consortium (IMGSAC) previously identified linkage loci on chromosomes 7 and 2, termed AUTS1 and AUTS5, respectively. In this study, we evaluated the patterns of linkage disequilibrium (LD) and the distribution of haplotype blocks, utilising data from the HapMap project, across the two strongest peaks of linkage on chromosome 2 and 7. More than 3000 SNPs have been selected in each locus in all known genes, as well as SNPs in non-genic highly conserved sequences. All markers have been genotyped to perform a high-density association analysis and to explore copy number variation within these regions. The study sample consisted of 127 and 126 multiplex families, showing linkage to the AUTS1 and AUTS5 regions, respectively, and 188 gender-matched controls. Association and CNV analysis implicated several new genes, including IMMP2L and DOCK4 on chromosome 7 and ZNF533 and NOSTRIN on the chromosome 2. Particularly, my contribution to this project focused on the characterization of the best candidate gene in each locus: On the AUTS5 locus I carried out a transcript study of ZNF533 in different human tissues to verify which isoforms and start exons were expressed. High transcript variability and a new exon, never described before, has been identified in this analysis. Furthermore, I selected 31 probands for the risk haplotype and performed a mutation screen of all known exons in order to identify novel coding variants associated to autism. On the AUTS1 locus a duplication was detected in one multiplex family that was transmitted from father to an affected son. This duplication interrupts two genes: IMMP2L and DOCK4 and warranted further analysis. Thus, I performed a screening of the cohort of IMGSAC collection (285 multiplex families), using a QMPSF assay (Quantitative Multiplex PCR of Short fluorescent Fragments) to analyse if CNVs in this genic region segregate with autism phenotype and compare their frequency with a sample of 475 UK controls. Evidence for a role of DOCK4 in autism susceptibility was supported by independent replication of association at rs2217262 and the finding of a deletion segregating in a sib-pair family. 2)Analysis of X chromosome inactivation. Skewed X chromosome inactivation (XCI) is observed in females carrying gene mutations involved in several X-linked syndromes. We aimed to estimate the role of X-linked genes in ASD susceptibility by ascertaining the XCI pattern in a sample of 543 informative mothers of children with ASD and in a sample of 164 affected girls. The study sample included families from different european consortia. I analysed the XCI inactivation pattern in a sample of italian mothers from singletons families with ASD and also a control groups (144 adult females and 40 young females). We observed no significant excess of skewed XCI in families with ASD. Interestingly, two mothers and one girl carrying known mutations in X-linked genes (NLGN3, ATRX, MECP2) showed highly skewed XCI, suggesting that ascertainment of XCI could reveal families with X-linked mutations. Linkage analysis was carried out in the subgroup of multiplex families with skewed XCI (≥80:20) and a modest increased allele sharing was obtained in the Xq27-Xq28 region, with a peak Z score of 1.75 close to rs719489. In this region FMR1 and MECP2 have been associated in some cases with austim and therefore represent candidates for the disorder. I performed a mutation screen of MECP2 in 33 unrelated probands from IMGSAC and italian families, showing XCI skewness. Recently, Xq28 duplications including MECP2, have been identified in families with MR, with asymptomatic carrier females showing extreme (>85%) skewing of XCI. For these reason I used the sample of probands from X-skewed families to perform CNV analysis by Real-time quantitative PCR. No duplications have been found in our sample. I have also confirmed all data using as alternative method the MLPA assay (Multiplex Ligation dependent Probe Amplification). 3)ASMT as functional candidate gene for autism. Recently, a possible involvement of the acetylserotonin O-methyltransferase (ASMT) gene in susceptibility to ASDs has been reported: mutation screening of the ASMT gene in 250 individuals from the PARIS collection revealed several rare variants with a likely functional role; Moreover, significant association was reported for two SNPs (rs4446909 and rs5989681) located in one of the two alternative promoters of the gene. To further investigate these findings, I carried out a replication study using a sample of 263 affected individuals from the IMGSAC collection and 390 control individuals. Several rare mutations were identified, including the splice site mutation IVS5+2T>C and the L326F substitution previously reported by Melke et al (2007), but the same rare variants have been found also in control individuals in our study. Interestingly, a new R319X stop mutation was found in a single autism proband of Italian origin and is absent from the entire control sample. Furthermore, no replication has been found in our case-control study typing the SNPs on the ASMT promoter B.