Attività scientifica

Pubblicazioni su riviste internazionali

Titolo: Assessment of Presentation Patterns, Clinical Severity and Extinction Modalities of Stereotypies in Autism: a crosssectional cohort study deemed the VICTORY project (2019)

Autore: Enzo Grossi, Elisa Caminada, Beatrice Vescovo, Tristana Castrignanò, Franco Vanzulli

Info: Submitted to Research in Developmental Disabilities


Lay Abstract 
Standardized video-recordings can help identify autistic repetitive behavior patterns, thus allowing for a better definition of major phenotypes.Twenty expert caregivers wearing a body cam recorded specific stereotypic behavior in a natural context during the everyday activities of 67 autistic subjects for 3 months of close follow-up. A team consisting of a senior child neuro-psychiatrist and a senior psychologist reviewed all the video recordings (1868) and selected 780 of them as the most significant to summarizing the whole 
spectrum within the given time window for each subject. Each video was further classified according to: components (motor, sensorial, vocal, intellective), complexity (2 classes, simple and complex), body parts involved (n=18) and sensory channels involved (hearing, sight, proprioception, taste, pain, smell). Subjects on study showed from 1 to 33 different patterns of repetitive behavior. The most frequent pattern represented a combination of simple motor and sensorial components (accounting for 23% of the total number) followed by simple motor and simple sensorial (9% and 8% respectively). Forty subjects expressed 
prevalent simple pattern and 27 prevalent complex patterns. No significant differences are found between the two groups according to autism severity.This study represents a first attempt to systematically document expression patterns of stereotyped behavior in a cohort of autistic subjects closely followed by professional educators. Open access to this video library as well as to the clinical data will be made available to interested researchers in the hope of better understanding this complex phenomenon and its correlation to specific clinical and demographic features. 

Scientific abstract 
Stereotypies, despite their high frequency and relevant diagnostic significance within autism, have yet to be fully elucidated because they occur within a broad spectrum of complex patterns. The aim of this study is to analyze stereotypies patterns by means of video-recordings. Twenty expert caregivers wearing a body cam recorded specific stereotypic behavior in a natural context during the everyday activities of 67 autistic subjects for 3 months of close follow-up. A team consisting of a senior child neuro-psychiatrist and a senior psychologist reviewed all the video recordings (1868) and selected 780 of them as the most significant to summarizing the whole spectrum within the given time window for each subject. Each video was further classified according to: components, complexity, body parts and sensory channels involved. The stereotypies spectrum for each subject ranged from 1 to 33 different patterns (average= 11.6; S.D.= 6.82). 
The most frequent pattern represented a combination of simple motor and sensorial components (accounting for 23% of the total number) followed by simple motor and simple sensorial (9% and 8% respectively). Forty subjects expressed prevalent simple pattern and 27 prevalent complex patterns. No significant differences are found between the two groups according to ADOS score severity. This study represents a first attempt to systematically document expression patterns of stereotyped behavior in a cohort of autistic subjects closely followed by professional educators.

Notes: Autism Research Unit, Villa Santa Maria Foundation

Titolo: Severe Bitter Taste Associated with Apremilast (2019)

Autore: G Damiani 1, 3 2, NL Bragazzi 4, E Grossi 5, S Petrou 6, D Radovanovic 7, M Rizzi 8, F Atzeni 9, P Sarzi-Puttini 10, P Santus 7, PD Pigatto 2, 3, C Franchi 2

Info: Dermatologic Therapy




1-Study Center of Young Dermatologists Italian Network (YDIN), GISED, Bergamo, Italy

2-Clinical Dermatology, IRCCS Istituto Ortopedico Galeazzi, Milan, Italy

3-Department of Biomedical, Surgical and Dental Sciences, University of Milan, Milan, Italy

4-School of Public Health, Department of Health Sciences (DISSAL), University of Genoa, Genoa, Italy

5-Villa Santa Maria Foundation, Tavernerio, Italy

6-St. George's University School of Medicine, Grenada, West Indies

7-Department of Biomedical and Clinicl, Sciences (DIBIC), University of Milan, Milan, Italy

8-Respiratory Unit "Luigi Sacco" University Hospital; ASST Fatebenefratelli-SaccoMilan, Italy

9-Rheumatology Unit, University of Messina, Italy 10-Rheumatology Unit, "Luigi Sacco" University Hospital; ASST Fatebenefratelli-Sacco, Milan, Italy

Titolo: Directional Relationship between Vitamin D Status and Prediabetes: a New Approach From Artificial Neural Network in a Cohort of Workers with Overweight-Obesity (2019)

Autore: a Luisella Vigna, b Amedea Silvia Tirelli, c Enzo Grossi, d Stefano Turolo, e Laura Tomaino, b Filomena Napolitano, f Massimo Buscema, a Luciano Riboldi

Info: Journal of the American College of Nutrition


Abstract: Objective 
Despite the increasing literature on the association of diabetes with inflammation, cardiovascular risk, and Vitamin D (25(OH)D) concentrations, strong evidence on the direction of causality between these factors is still lacking. This gap could 
be addressed by means of artificial neural networks (ANN) analysis 
Retrospective observational study was carried out by means of an innovative data mining analysis –known as auto-contractive map (AutoCM)-, and semantic mapping 
followed by Activation and Competition System on data of workers referring to an occupational-health outpatient clinic Parameters analyzed included weight, height, waist circumference, BMI, percentage of fat mass, glucose, insulin, HbA1c, creatinine, total cholesterol, low and high density lipoprotein, triglycerides, uric acid, fibrinogen, homocysteine, CRP, diastolic and systolic blood pressure, and 25(OH)D. 
The study included 309 workers. Of these 23.6% were overweight or obese with rates of I, II and III level-obesity, respectively of 40.5%, 23.3% and 12.6%. All mean biochemical values were in normal range, except for T-Chol, HDL, LDL, CRP, and 
25(OH)D. HbAC1 was between 39-46 mmol/mol in 51.78%. 25(OH)D levels were sufficient in only 12.6%. Highest inverse correlation for hyperglycemia onset was with BMI and waste circumference, suggesting a protective role of 25(OH)D against their increase. Auto- CM processing and the semantic map evidenced direct association of 25(OH)D with high link strength (0.99) to low CRP levels and low HDL levels. Low 25(OH)D lead to changes in glucose which affected metabolic syndrome biomarkers, first of which HOMA index and blood glucose insulin, but not 25(OH)D. 
The use of ANN suggests a key role of 25(OH)D respect to all considered metabolic parameters in the development of diabetes and evidences a causation between low 25(OH)D and high glucose concentrations.


a Department of Preventive Medicine, Occupational Health Unit, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milan, Italy

b Department of Clinical Chemistry and Microbiology, Bacteriology and Virology Units; Ospedale Maggiore Policlinico, Milan, ITALY

c Villa Santa Maria Foundation, Tavernerio, Italy

d Pediatric Nephrology & Dialysis, Milano Fondazione IRCCS Cà Grande Ospedale Maggiore Policlinico, University of Milan, 20122 Milan, Italy

e Pediatric Intermediate Care Unit, Department of Clinical and Community Health Sciences (DISCCO), Fondazione IRCCS Ospedale Cà Granda-Ospedale Maggiore Policlinico, University of Milan, 20122 Milan, Italy

f Semeion Research Centre of Sciences of Communication, Rome, Italy and University of Colorado, Denver, CO, USA

Titolo: Motor skills as moderators of core symptoms in Autism Spectrum Disorders: preliminary data from an exploratory analysis with Artificial Neural Networks (2018)

Autore: Francesca Fulceri 1, Enzo Grossi 2, Annarita Contaldo 1, Antonio Narzisi 1, Fabio Apicella 1, Ilaria Parrini 1, Raffaella Tancredi 1, Sara Calderoni 1, 3*, Filippo Muratori 1, 3

Info: Frontiers


Motor disturbances have been widely observed in children with autism spectrum disorder (ASD), and motor problems are currently reported as associated features supporting the diagnosis of ASD in the current Diagnostic and Statistical Manual of Mental Disorders (DSM-5). Studies on this issue reported disturbances in different motor domains, including both gross and fine motor areas as well as coordination, postural control, and standing balance. However, they failed to clearly state whether motor impairments are related to demographical and developmental features of ASD. Both the different methodological approaches assessing motor skills and the heterogeneity in clinical features of participants analyzed have been implicated as contributors to variance in findings. However, the non-linearity of the relationships between variables may account for the inability of the traditional analysis to grasp the core problem suggesting that the “single symptom approach analysis” should be overcome.
Artificial neural networks (ANNs) are computational adaptive systems inspired by the functioning processes of the human brain particularly adapted to solving non-linear problems. This study aimed to apply the ANNs to reveal the entire spectrum of the relationship between motor skills and clinical variables. 32 male children with ASD [mean age: 48.5 months (SD: 8.8); age range: 30-60 months] were recruited in a tertiary care university hospital. A multidisciplinary comprehensive diagnostic evaluation was associated with a standardized assessment battery for motor skills, the Peabody Developmental Motor Scale-Second Edition.
Exploratory analyses were performed through the ANNs. The findings revealed that poor motor skills were a common clinical feature of preschoolers with ASD, relating both to the high level of repetitive behaviors and to the low level of expressive language. Moreover, unobvious trends among motor, cognitive and social skills have been detected.
In conclusion, motor abnormalities in preschoolers with ASD were widespread, and the degree of impairment may inform clinicians about the severity of ASD core symptoms. Understanding motor disturbances in children with ASD may be relevant to clarify neurobiological basis and ultimately to guide the development of tailored treatments.


1 Fondazione Stella Maris (IRCCS), Italy,
2 Autism Research Unit, Villa Santa Maria Scs, Italy,
3 Università degli Studi di Pisa, Italy

Titolo: Artificial neural networks help to identify disease subsets and to predict lymphoma in primary Sjögren’s syndrome (2018)

Autore: Chiara Baldini1, Francesco Ferro1, Nicoletta Luciano1, Stefano Bombardieri1, Enzo Grossi2

Info: Clinical and Experimental Rheumatology 2018, 36 (Suppl. 112): S137-S144



Abstract: Objective. Primary Sjögren’s syndrome (pSS) is a complex chronic systemic disorder, for which specific and effective therapeutic interventions are still lacking. In this era of precision medicine, there is a clear need for a better definition of disease phenotypes to foster the research of novel specific biomarkers and new therapeutic targets. 
The main objectives of this work are:1) to compare Auto Contractive Map (AutoCM), a data mining tool based on an artificial neural network (ANN) versus conventional Principal Component Analysis (PCA) in discriminating different pSS subsets and 2) to specifically focus on variables predictive of MALT-NHL development, assessing the previsional gain of the predictive models developed. 
Methods. Out of a historic cohort of 850 patients, we selected 542 cases of pSS fulfilling the AECG criteria 2002. Thirty-seven variables were analysed including: patient demographics, glandular symptoms, systemic features, biological abnormalities and MALT-NHLs. AutoCM was used to compute the association of strength of each variable with all other variables in the dataset. PCA was applied to the same data set. 
Results. Both PCA and AutoCM confirmed the associations between autoantibody positivity and several pSS clinical manifestations, highlighting the importance of serological biomarkers in pSS phenotyping. However, AutoCM allowed us to clearly distinguish pSS patients presenting with predominant glandular manifestations and no or mild extra-glandular features from those with a more severe clinical presentation. 
Out of 542 patients, we had 27 cases of MALT-NHLs. The AutoCM highlighted that, besides other traditional lymphoproliferative risk factors (i.e. salivary gland enlargement, low C4, leukocytopenia, cryoglobulins, monoclonal gammopathy, disease duration), rheumatoid factor was strongly associated to MALT-NHLs development. By applying data mining analysis, we obtained a predictive model characterised by a sensitivity of 92.5% and a specificity of 98%. If we restricted the analysis to the seven most significant variables, the sensitivity of the model was 96.2% and its specificity 96%. 
Conclusion. Our study has shed new light on the possibility of using novel tools to extract hidden, previously unknown and potentially useful information in complex diseases like pSS, facing the challenge of disease phenotyping as a prerequisite for discovering novel specific biomarkers and new therapeutic targets.

Notes: 1.Rheumatology Unit, University of Pisa

2.Villa Santa Maria Foundation, Tavernerio, Italy

Titolo: Artificial Neural Networks (ANNs) as a Reliable Tool for the Assessment of Fracture Risk in Postmenopausal Women (2018)

Autore: Gloria Bonaccorsi1, Carlo Cervellati2, Enzo Grossi3, Enrica Fila1, Leo Massari4, Nicola Veronese5, Francesco Pio Cafarelli6, Giuseppe Guglielmi6, 7*

Info: British Journal of Research ISSN 2394-3718



Abstract: Artificial neural networks (ANNs) are a computational tool, based on highly non-linear mathematics models with potential applications in the prediction of osteoporotic fractures. 
Therefore, the present study aimed to evaluate the potential of ANNs analysis in the prediction of bone fragility fractures in post-menopausal women. ANNs prognostic performance in identifying vertebral morphometric deformity was compared with that of the widely used tool FRAX® in a sample of 587 Caucasian postmenopausal women underwent densitometry and morphometric analyses for the detection of vertebral fractures. 
The analysis of areas under the curve (AUCs) showed that sensitivity for ANNs (74%) almost doubled that found for FRAX ® (38%), with the latter presenting a specificity higher than the proposed tool (96 vs. 77%). Overall, ANN-based analysis was able to highlight high-risk patients with a global higher accuracy (74%) compared to that obtained by FRAX (67%). 
In conclusion, our data showed that compared to WHO’s algorithm ANNs had higher sensitivity in identifying vertebral deformity, thus suggesting a “promising role” in the prediction of osteoporotic fracture in postmenopausal women. However, further studies on larger sample are needed to definitely establish the clinical reliability of ANNs.

Notes: 1.Department of Morphology, Surgery and Experimental Medicine, Menopause and Osteoporosis Centre, University of Ferrara, Via Boschetto 29, 44124, Ferrara, Italy

2.Department of Biomedical and Specialist Surgical Sciences, Section of Medical Biochemistry, Molecular Biology and Genetics, University of Ferrara, Via Borsari 46, 44121, Ferrara, Italy

3.Villa Santa Maria Foundation, Via IV Novembre, 22038 Tavernerio (CO), Italy

4.Department of Morphology, Surgery and Experimental Medicine, Section of Orthopedic Clinic, University of Ferrara, Via Aldo Moro 8, 44124, Cona, Ferrara, Italy

5.National Research Council, Neuroscience Institute, Aging Branch, via Giustiniani, 2, 35128, Padova, Italy

6.Department of Clinical and Experimental Medicine, Foggia University School of Medicine, Via L. Pinto, 1, Foggia, Italy

7.Department of Radiology, Scientific Institute Hospital “Casa Sollievo della Sofferenza”, San Giovanni Rotondo, Italy

Titolo: Plasma Fatty Acid Profile as Biomarker of Coronary Artery Disease: a Pilot Study Using Fourth Generation Artificial Neural Networks (2018)

Autore: E Dozio*, E Vianello*, E Grossi**, L Menicanti***, G Schmitz****, M M Corsi Romanelli*, *****

Info: Journal of Biological Regulators & Homeostatic Agents, Vol. 32, no. 4, 79-85 (2018)


Abstract: Many studies, focused on identifying new biomarkers for coronary artery disease (CAD) risk computation and monitoring, suggested a potential diagnostic role for fatty acids (FA). In the present study, we explored the potential diagnostic role of FA by using a data mining approach based on fourth generation artificial neural networks (ANN). Forty-one male subjects were enrolled. According to coronary angiography, 31 displayed CAD and 10 did not (non-CAD, control group). FA analysis was performed on plasma samples using a gas chromatography-mass spectrometry system and analyses were performed by an ANN method. The variables most closely related to CAD were low levels of alpha-linolenic acid, eicosapentaenoic acid, eicosatetraenoic and docosahexaenoic acids. High levels of 1,1-dimethoxyhexadecane, total dimethyl acetals and docosatetraenoic acid were related to non-CAD condition. This subset of variables, which were most closely correlated to the target diagnosis, achieved a consistent predictive rate. The average accuracy obtained was 76.5%, with 93% of sensitivity and 60% of specificity. The area under the ROC curve was equal to 0.79. In conclusion, our study highlighted the association between different plasma FA species, CAD and non-CAD conditions. The specific subset of variables could be of interest as a new diagnostic tool for CAD management.


Notes: *Department of Biomedical Sciences for Health, Università degli Studi di Milano, Milan, Italy

**Villa Santa Maria Institute, Tavernerio, Como, Italy

***Department of Cardiac Surgery, I.R.C.C.S. Policlinico San Donato, San Donato Milanese, Milan, Italy

****Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Regensburg, Germany

*****Service of Laboratory Medicine 1-Clinical Pathology, I.R.C.C.S. Policlinico San Donato, San Donato Milanese, Milan, Italy

Titolo: Toe walking assessment in autism spectrum disorder subjects: a systematic review (2018)

Autore: Giulio Valagussa*,**, Luca Trentin*, Alessio Signori***, Enzo Grossi*

Info: Accepted by Autism Research


Abstract: There is increasing evidence that autism spectrum disorder (ASD) subjects have also motor impairments. Toe walking (TW) is a phenomenon that can be found in ASD subjects during gait, even if this condition was found not to be necessarily related only to walking, since these children often also stand and run on their tiptoes. Since persistent TW in ASD subjects may contribute to secondary shortening of the Achilles’s tendon, it becomes important to have an assessment tool and/or outcome measure for both the clinical and rehabilitative settings. The aim of this systematic review is to critically evaluate and describe the methods employed to assess toe-walking in ASD subjects. 
The systematic review protocol was previously registered on PROSPERO. We conducted an extensive literature search in PubMed, CINAHL, PsycINFO, The Cochrane Library, and Scopus databases. There were no restrictions on the types of study design eligible for inclusion. Ten studies were included in the systematic review. Risk of bias of the included studies was conducted using the following instruments depending on the study types: STROBE Statement, Cochrane risk of bias tool, and CARE checklist. Almost all the included studies (8/10) proposed a TTB assessment only during walking. Nine out of ten of the included studies assessed TTB using a qualitative methodology. 
The results evidenced the heterogeneity of qualitative methods and a lack of a structured quantitative test to assess toe walking 
in ASD subjects.

Notes: *Autism Research Unit, Villa Santa Maria Foundation, Tavernerio (CO), Italy

**School of Medicine and Surgery – University of Milano Bicocca, Milan, Italy

***Department of Health Sciences (DISSAL), Section of Biostatistics, University of Genoa, Genoa, Italy

Titolo: Magic Moments: Determinants of Stress Relief and Subjective Wellbeing from Visiting a Cultural Heritage Site (2018)

Autore: Enzo Grossi (1), Giorgio Tavano Blessi (2), Pier Luigi Sacco (2, 3, 4)

Info: Culture, Medicine and Psychiatry



Abstract: We provide an experimental evaluation of the impact of aesthetic experiences in terms of stress reduction (cortisol levels) and wellbeing increase. The test experience is a visit to the vault of the Sanctuary of Vicoforte, Italy. Data have been collected using a double step method. A structured interview in relation to the individual subjective well-being has been administered to a sample of 100 subjects. 
In addition, a sample of their saliva has been taken, and its cortisol level measured, before and after the experience, and likewise for momentary wellbeing measured on a Visual Analogous Scale. Subjects reported an average increase of 40% in wellbeing and a decrease of the 60% in the cortisol level. The recorded cortisol level values dropped on average well beyond the decrease normally associated to its circadian cycle. The modulating role of various variables has been appreciated, and profiling of the typical subjects who are wellbeing respondents/non-respondents and cortisol respondents/non-respondents has been carried out. 
We conclude that aesthetic experience seems to have a noticeable impact on individual physical and mental health. In both dominions, cultural participation intensity is significantly correlated to the response. The study underlines the potential of the arts and culture as a new platform for public health practices and new approaches to welfare policy design.

Notes: 1 Villa Santa Maria Institute, Tavernerio, Italy 2 IULM University, Milan, Italy 3 FBK-IRVAPP, Trento, Italy 4 Harvard University and MetaLAB (at) Harvard, Cambridge, MA, USA

Titolo: Determinants of metabolic syndrome in obese workers: gender differences in perceived job-related stress and in psychological characteristics identified using artificial neural networks (2018)

Autore: Luisella Vigna (1), Amelia Brunani (2), Agostino Brugnera (3), Enzo Grossi (4), Angelo Compare (3), Amedea S. Tirelli (5), Diana M. Conti (1), Gianna M. Agnelli (1), Lars L. Andersen (6, 7), Massimo Buscema (8, 9), Luciano Riboldi (1)

Info: Eating and Weight Disorders - Studies on Anorexia, Bulimia and Obesity



Objective: The metabolic syndrome (MS) is a multifactorial disorder associated with a higher risk of developing cardiovascular diseases and type 2 diabetes. However, its pathophysiology and risk factors are still poorly understood. In this study, we investigated the associations among gender, psychosocial variables, job-related stress and the presence of MS in a cohort of obese Caucasian workers. 

Methods: A total of 210 outpatients (142 women, 68 men) from an occupational medicine service was enrolled in the study. Age, BMI, waist circumference, fasting glucose, blood pressure, triglycerides and HDL cholesterol were collected to define MS. In addition, we evaluated eating behaviors, depressive symptoms, and work-related stress. Data analyses were performed with an artificial neural network algorithm called Auto Semantic Connectivity Map (AutoCM), using all available variables. 

Results: MS was diagnosed in 54.4 and 33.1% of the men and women, respectively. AutoCM evidenced gender-specific clusters associated with the presence or absence of MS. Men with a moderate occupational physical activity, obesity, older age and higher levels of decision-making freedom at work were more likely to have a diagnosis of MS than women. Women with lower levels of decision-making freedom, and higher levels of psychological demands and social support at work had a lower incidence of MS but showed higher levels of binge eating and depressive symptomatology. 

Conclusion: We found a complex gender-related association between MS, psychosocial risk factors and occupational determinants. The use of these information in surveillance workplace programs might prevent the onset of MS and decrease the chance of negative long-term outcomes. 

Level of evidence Level V, observational study.


Notes: 1 Department of Preventive Medicine, Occupational Health Unit, Clinica del Lavoro Luigi Devoto, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico 2 Rehabilitation Medicine, IRCCS Istituto Auxologico Italiano, S. Giuseppe Hospital, Verbania, Italy 3 Department of Human and Social Sciences, University of Bergamo, Bergamo, Italy 4 Villa Santa Maria Foundation, Tavernerio, Italy 5 Laboratory of Clinical Chemistry and Microbiology, Fondazione IRCCS Ca’ Granda, Ospedale Maggiore Policlinico, Milan, Italy 6 National Research Centre for the Working Environment, Copenhagen, Denmark 7 Sport Sciences, Department of Health Science and Technology, Aalborg University, Aalborg, Denmark 8 Semeion Research Centre of Sciences of Communication, Rome, Italy 9 University of Colorado, Denver, CO, USA

Titolo: Current Knowledge on Endocrine Disrupting Chemicals (Edcs) from Animal Biology to Humans, From Pregnancy to Adulthood: Highlights from A National Italian Meeting (2018)

Autore: Maria Elisabeth Street *, Sabrina Angelini, Sergio Bernasconi, Ernesto Burgio, Alessandra Cassio, Cecilia Catellani, Francesca Cirillo, Annalisa Deodati, Enrica Fabbrizi, Vassilios Fanos, Giancarlo Gargano, Enzo Grossi, Lorenzo Iughetti, Pietro Lazzeroni

Info: International Journal of Molecular Sciences


Wildlife has often presented and suggested the effects of endocrine disrupting chemicals (EDCs). Animal studies have given us an important opportunity to understand the mechanisms of action of many chemicals on the endocrine system and on neurodevelopment and behaviour, and to evaluate the effects of doses, time and duration of exposure. Although results are sometimes conflicting because of confounding factors, epidemiological studies in humans suggest effects of EDCs on prenatal growth, thyroid function, glucose metabolism and obesity, puberty, fertility, and on carcinogenesis mainly through epigenetic mechanisms. This manuscript reviews the reports of a multidisciplinary national meeting on this topic. 

Titolo: Pregnancy risk factors related to autism: an Italian case-control study in mothers of children with ASD, their siblings and of typically developing children (2018)

Autore: Enzo Grossi, Lucia Migliore, Filippo Muratori

Info: Journal of Developmental Origins of Health and Disease, 23 April 2018


This study, carried out in two Italian Institutions, assesses the frequency of 27 potential autism risk factors related to pregnancy and peri- and postnatal periods by interviewing mothers who had children with autism, children with autism and one or two typically developing siblings, or only typically developing children. The clinical sample included three case groups: 73 children and adolescents with autism (Group A), 35 children and adolescents with autism (Group A1) having 45 siblings (Group B) and 96 typically developing children (Group C) matched for gender and age. Twenty-five out of 27 of risk factors presented a higher frequency in Group A in comparison with Group C and for nine of them a statistically significant difference was found. Twenty-one out of 27 of risk factors presented a higher frequency in Group A in comparison with Group B. A higher prevalence of environmental risk factors was observed in 11 risk factors in the Group A1 in comparison with Group B and for nine of them an odds ratio higher than 1.5 was found. For 13 factors there was a progressive increase in frequency going from Group C, B and A and a statistically higher prevalence of the mean number of stressful events per pregnancy was recorded in Group A when compared with Groups B and C. The results suggest that environmental, incidental phenomena and stressful life events can influence pregnancy outcome in predisposed subjects, pointing out a possible threshold effect in women who are predisposed to have suboptimal pregnancies.

Titolo: The role of carboxy-terminal cross-linking telopeptide of type I collagen, dual x-ray absorptiometry bone strain and Romberg test in a new osteoporotic fracture risk evaluation: A proposal from an observational study. (2018)

Autore: Fabio M Ulivieri, Luca P Piodi, Enzo Grossi, Luca Rinaudo, Carmelo Messina, Anna P Tassi, Marcello Filopanti, Anna Tirelli, Francesco Sardanelli

Info: Plos One January 5, 2018



The consolidated way of diagnosing and treating osteoporosis in order to prevent fragility fractures has recently been questioned by some papers, which complained of overdiagnosis and consequent overtreatment of this pathology with underestimating other causes of the fragility fractures, like falls. A new clinical approach is proposed for identifying the subgroup of patients prone to fragility fractures. This retrospective observational study was conducted from January to June 2015 at the Nuclear Medicine-Bone Metabolic Unit of the of the Fondazione IRCCS Ca' Granda, Milan, Italy. An Italian population of 125 consecutive postmenopausal women was investigated for bone quantity and bone quality. Patients with neurological diseases regarding balance and vestibular dysfunction, sarcopenia, past or current history of diseases and use of drugs known to affect bone metabolism were excluded. Dual X-ray absorptiometry was used to assess bone quantity (bone mineral density) and bone quality (trabecular bone score and bone strain). Biochemical markers of bone turnover (type I collagen carboxy-terminal telopeptide, alkaline phosphatase, vitamin D) have been measured. Morphometric fractures have been searched by spine radiography. Balance was evaluated by the Romberg test. The data were evaluated with the neural network analysis using the Auto Contractive Map algorithm. The resulting semantic map shows the Minimal Spanning Tree and the Maximally Regular Graph of the interrelations between bone status parameters, balance conditions and fractures of the studied population. A low fracture risk seems to be related to a low carboxy-terminal cross-linking telopeptide of type I collagen level, whereas a positive Romberg test, together with compromised bone trabecular microarchitecture DXA parameters, appears to be strictly connected with fragility fractures. A simple assessment of the risk of fragility fracture is proposed in order to identify those frail patients at risk for osteoporotic fractures, who may have the best benefit from a pharmacological and physiotherapeutic approach.

Titolo: The Role of Trabecular Bone Score and Hip Geometry in Thalassemia Major: A Neural Network Analysis (2017)

Autore: Marina Baldini, Enzo Grossi, Maria Domenica Cappellini, Carmelo Messina, Alessia Marcon, Elena Cassinerio, Lorena Airaghi, Giuseppe Guglielmi and Fabio Massimo Ulivieri*

Info: British Journal of Research ISSN 2394-3718


Osteopathy in thalassemia is a very heterogeneous condition; severity depends on multiple factors, interacting through nonlinear mechanisms. Classic statistics have limitations when applied to the study of such highly complex relationships. Currently, an alternative approach of analysis is represented by the artificial neural networks (ANNs), powerful mathematical tools, increasingly applied to analyze multifactorial databases, as considered more appropriate than classic statistics. We adopted this specialized mathematical method to 76 thalassemia major (TM) patients. In all of them dual energy X-ray absorptiometry (DXA) was performed to measure bone mineral density, and two recent developments were included: trabecular bone score, evaluating bone microarchitecture, and hip structural analysis, evaluating hip geometry. The relationships between bone status and endocrine, hematologic, and clinical parameters were investigated. Using a particular ANN (Auto Contractive Map algorithm), the strength of inter-variable association was defined and a connectivity map generated, visually representing the main connections among the entered variables. Iron status indices (ferritin, liver iron concentration) emerged as the most important variables, dividing the map into two sectors, with parameters indicating satisfactory bone condition in the upper, those indicating poor condition in the lower, near the variable “fractures”. The Auto Contractive Map highlighted the key role of bone quantity, bone geometry, and microarchitecture in defining thalassemic bone condition. Among numerous available indices, high femoral bone mineral density and low cross-sectional moment of inertia emerged as the gold standard to classify thalassemic patients for prognostic and therapeutic purposes.

Titolo: Role of the Human Breast Milk-Associated Microbiota on the Newborns’ Immune System: A Mini Review (2017)

Autore: Marco Toscano, Roberta De Grandi, Enzo Grossi and Lorenzo Drago*

Info: Frontiers in Microbiology, 25 October 2017



The human milk is a rich and complete nourishment that is essential for the correct development of the infant’s organism (Ballard and Morrow, 2013). The first milk produced by mothers after the delivery is called colostrum and it is biochemically and functionally different from the mature milk (Castellote et al., 2011). Colostrum, indeed, contains high concentration of lactoferrin, Immunoglobulin A (IgA), leukocytes and specific developmental factors, and a low amount of lactose, potassium and calcium, underlying its immunological functions rather than nutritional (Kulski and Hartmann, 1981; Pang and Hartmann, 2007). From 5 days to 2 weeks postpartum, there is the production of transitional milk which shares some characteristics of colostrum, although its main function is to support newborns at nutritional level (Henderson et al., 2008; Nommsen-Rivers et al., 2012). Finally, 2 weeks after the delivery the milk can be considered as mature and its composition tends to be stable over the time, even if slight variations can occur during lactation (Ballard and Morrow, 2013). The main components of human milk are: (i) macronutrients, such as protein, fat and lactose, which concentration depends on the stage of lactation and maternal characteristics; (ii) micronutrients, including vitamins A, B1, B2, B12 and D that vary in human milk in relation to maternal diet and body stores; (iii) growth factor, which are strongly active on the endocrine system, nervous system, vasculature and intestinal tract; (iv) immunological factors, which are essential to defend the newborn from inflammation and infection, and for this reason, the early milk is rich in immune components that can support infants in the first delicate stages of their life; (v) the microbiota, which comprises more than 200 different bacterial species with a pivotal role in the formation of the newborn’s first gut microbiota (Drago et al., 2017). The aim of the present Mini Review is to highlight the specific and fundamental role of human milk-associated bacteria in modulating and influencing the newborns’ immune system during their life.

Titolo: Impact of delivery mode on the colostrum microbiota composition (2017)

Autore: Marco Toscano, Roberta De Grandi, Diego Giampietro Peroni, Enzo Grossi, Valentina Facchin, Pasquale Comberiati and Lorenzo Drago.

Info: BMC Microbiology (2017) 17:205



Background Breast milk is a rich nutrient with a temporally dynamic nature. In particular, numerous alterations in the nutritional, immunological and microbiological content occur during the transition from colostrum to mature milk. The objective of our study was to evaluate the potential impact of delivery mode on the microbiota of colostrum, at both the quantitative and qualitative levels (bacterial abundance and microbiota network). Methods Twenty-nine Italian mothers (15 vaginal deliveries vs 14 Cesarean sections) were enrolled in the study. The microbiota of colostrum samples was analyzed by next generation sequencing (Ion Torrent Personal Genome Machine). The colostrum microbiota network associated with Cesarean section and vaginal delivery was evaluated by means of the Auto Contractive Map (AutoCM), a mathematical methodology based on Artificial Neural Network (ANN) architecture. Results Numerous differences between Cesarean section and vaginal delivery colostrum were observed. Vaginal delivery colostrum had a significant lower abundance of Pseudomonas spp., Staphylococcus spp. and Prevotella spp. when compared to Cesarean section colostrum samples. Furthermore, the mode of delivery had a strong influence on the microbiota network, as Cesarean section colostrum showed a higher number of bacterial hubs if compared to vaginal delivery, sharing only 5 hubs. Interestingly, the colostrum of mothers who had a Cesarean section was richer in environmental bacteria than mothers who underwent vaginal delivery. Finally, both Cesarean section and vaginal delivery colostrum contained a greater number of anaerobic bacteria genera. Conclusions The mode of delivery had a large impact on the microbiota composition of colostrum. Further studies are needed to better define the meaning of the differences we observed between Cesarean section and vaginal delivery colostrum microbiota.

Titolo: Effects of Improvisational Music Therapy vs Enhanced Standard Care on Symptom Severity Among Children With Autism Spectrum Disorder (2017)

Autore: Łucja Bieleninik (1), Monika Geretsegger (1), Karin Mössler (1), Jörg Assmus (1), Grace Thompson (2), Gustavo Gattino (3,4), Cochavit Elefant (5), Tali Gottfried (6), Roberta Igliozzi (7), Filippo Muratori (7,8), Ferdinando Suvini (7), Jinah Kim (9), Mike

Info: JAMA - The Journal of the American Medical Association, 2017; 318(6): 525-535



Importance Music therapy may facilitate skills in areas affected by autism spectrum disorder (ASD), such as social interaction and communication. Objective To evaluate effects of improvisational music therapy on generalized social communication skills of children with ASD. Design, Setting, and Participants Assessor-blinded, randomized clinical trial, conducted in 9 countries and enrolling children aged 4 to 7 years with ASD. Children were recruited from November 2011 to November 2015, with follow-up between January 2012 and November 2016. Interventions Enhanced standard care (n = 182) vs enhanced standard care plus improvisational music therapy (n = 182), allocated in a 1:1 ratio. Enhanced standard care consisted of usual care as locally available plus parent counseling to discuss parents’ concerns and provide information about ASD. In improvisational music therapy, trained music therapists sang or played music with each child, attuned and adapted to the child’s focus of attention, to help children develop affect sharing and joint attention. Main Outcomes and Measures The primary outcome was symptom severity over 5 months, based on the Autism Diagnostic Observation Schedule (ADOS), social affect domain (range, 0-27; higher scores indicate greater severity; minimal clinically important difference, 1). Prespecified secondary outcomes included parent-rated social responsiveness. All outcomes were also assessed at 2 and 12 months. Results Among 364 participants randomized (mean age, 5.4 years; 83% boys), 314 (86%) completed the primary end point and 290 (80%) completed the last end point. Over 5 months, participants assigned to music therapy received a median of 19 music therapy, 3 parent counseling, and 36 other therapy sessions, compared with 3 parent counseling and 45 other therapy sessions for those assigned to enhanced standard care. From baseline to 5 months, mean ADOS social affect scores estimated by linear mixed-effects models decreased from 14.08 to 13.23 in the music therapy group and from 13.49 to 12.58 in the standard care group (mean difference, 0.06 [95% CI, −0.70 to 0.81]; P = .88), with no significant difference in improvement. Of 20 exploratory secondary outcomes, 17 showed no significant difference. Conclusions and Relevance Among children with autism spectrum disorder, improvisational music therapy, compared with enhanced standard care, resulted in no significant difference in symptom severity based on the ADOS social affect domain over 5 months. These findings do not support the use of improvisational music therapy for symptom reduction in children with autism spectrum disorder. Trial Registration Identifier: ISRCTN78923965

Titolo: Diagnosis of autism through EEG processed by novel artificial adaptive systems: a proof of concept pilot study (2017)

Autore: Enzo Grossi, Chiara Olivieri & Massimo Buscema

Info: Computer Methods and Programs in Biomedicine 142 (2017) 73–79



Background MS-ROM/I-FAST is a new, complex algorithm used for blind classification of the original EEG tracing of each subject. This is accomplished by recording and analyzing a few minutes of their EEG without artifacts removing. A proof of concept study well equipped with cross-validation techniques previously published showed accuracy values ranging from 94%-98% in discerning subjects with Mild Cognitive Impairment and/or Alzheimer’s Disease from healthy elderly people. Since ASD is known to show deviant patterns in simple resting state EEG recordings, this supports the idea that the atypical organization of the cerebral cortex present in autism should result in an EEG signature open to detection through potent analytical systems like Artificial Neural networks (ANNs.) Aim of the study The aim of the study is to assess how effective this methodology distinguishes ADS subjects from typically developing ones. Methods Fifteen definite ASD subjects (13 males; 2 females; age range 7-14; mean value = 10.4) and ten typically developing subjects ( 4 males; 6 females; age range 7-12; mean value 9.2 ) were included in the study. Patients received independent Autism diagnoses according to DSM-V criteria, then subsequently confirmed by a qualified psychiatrist at Villa Santa Maria, where the patients reside, using the ADOS scale (overall severity score had a range from a minimum of 4 to a maximum of 10 points, average = 7.9). No autistic child was affected by genetic conditions and/or cerebral malformations documented by neuroimaging and epilepsy. A continuous segment of artefact-free EEG data lasting 60 s was used to compute multi-scale entropy values and for subsequent analyses. A Multi-scale ranked organizing map (MS-ROM), based on the self-organizing map (SOM) neural network, coupled with the TWIST system (an evolutionary system able to select predictive features) created an invariant features vector input of EEG on which supervised machine learning systems acted as blind classifiers. Results After MS-ROM/I-FAST preprocessing, the overall predictive capability of different machine learning systems in deciphering autistic cases from normal ones resulted extremely good reaching also perfect distinction with specific classifiers. These results were obtained at different times in separate experiments performed on the same training and testing subsets. The similarities among the ANN weight matrixes measured with apposite algorithms were not affected by the age of the subjects. This suggests that the ANNs do not read age-related EEG patterns, but rather invariant features related to the brain’s underlying disconnection signature. Conclusion This pilot study seems to open up new avenues for the development of non-invasive diagnostic testing for the early detection of ASD.rumors and disappointments at media level, while the third problem is specifically felt by Pharmaceutical Companies and Institutional Review Boards releasing the clearance for first in man trials.

Titolo: Assessment of presentation patterns, clinical severity and sensorial mechanism of tip-toe behavior in ASD children: a cohort observational study (2017)

Autore: Giulio Valagussa, Luca Trentin, Valeria Balatti, Enzo Grossi

Info: Autism Research 2017



We assessed presentation patterns, clinical severity and sensorial mechanism of tip-toe behavior (TTB ), more commonly known as toe walking , in a cohort of ASD subjects in two studies. The first study includes 69 consecutive ASD subjects (57 males, 12 females, mean age = 14 years – 3.7 SD) under observation at our institute. A therapist assessed the presence of TTB during standing, walking and running using direct observation and an interview of the main caregiver living with the children was conducted. The prevalence of TTB was 32%. We found three clinical presentation patterns of TTB: 1) present while standing, walking and running (45.5%), 2) present when walking and running (18.4%) or 3) present only when running (36.4%). TTB subjects were more frequently non-verbal than subjects without TTB (72.7% vs 44.6% - p= 0.03). On the other hand, no significant difference in ASD severity according to the ADOS scale was found between TTB and non-TTB subjects. In the second study, carried out in a subgroup of 14 ASD subjects (7 TTB and 7 non-TTB), we show that acting upon a soft floor surface (foam mats) made a substantial difference in reducing the phenomenon. TTB is a frequent phenomenon in individuals with ASD and may occur in three mutually exclusive modalities, which ultimately defines what is commonly known as toe walking. The presence of TTB seems correlated to the severity of language delay. Foot contact on soft surfaces reduces TTB both during static and/or dynamic tasks. Further evaluation is needed to clarify the potential pathophysiological implications of this phenomenon.

Titolo: Inequalities in access to biological treatments for psoriasis: Results from the Italian Psocare Registry (2016)

Autore: Luigi Naldi, Simone Cazzaniga, Marco Di Mercurio, Enzo Grossi, Antonio Addis

Info: British Journal of Dermatology Accepted manuscript online: 14 December 2016



Background: Limited evidence is available on the impact of socioeconomic factors in drug prescriptions for psoriasis. Objectives: To investigate factors influencing prescription of conventional versus biological treatment for psoriatic patients, based on the Psocare registry with a special focus on socioeconomic factors. Methods: This was a cross-sectional study evaluating the baseline data of patients included in the Italian Psocare Registry. All of the consecutive adult patients with a diagnosis of chronic plaque psoriasis (Ps) or psoriatic arthritis and who were prescribed a systemic treatment for Ps at the participating centres were included in this study. Univariate and multivariate analyses of the baseline factors associated with a biologics prescription were performed. Results: From September 2005 to September 2009, 12,838 patients were identified. A multivariate analysis revealed that, among other factors, completing a level of education higher than lower secondary school and being employed as a manager or a professional were independent factors associated with a biologics prescription at entry in the registry. Additional analyses on the association between these two variables and a severe Ps condition (i.e., psoriasis area and severity index [PASI] score > 20) revealed a significant increasing trend of severe disease towards lower educational attainment, while unemployed patients were more likely to have a more severe condition as compared to the other categories of workers. Conclusions: We documented inequalities of drug prescriptions for Ps in Italy, with a trend towards a higher frequency of prescription for more expensive biologics in higher socio-economic sectors of the population.

Titolo: Monitoring food selectivity in a group of children with autism spectrum disorders with direct observation: a comparative observational feasibility study (2016)

Autore: Enzo Grossi, Sara Melli, Marina Norsi

Info: Scientific Times Journal of Paediatrics, Volume 1 Issue 1



Food selectivity is a fairly common issue with children who have an autism spectrum disorder (ASD)’. Few studies based on direct observation of feeding behavior in youths with ASD are available in the literature.In this study we explore and monitor food selectivity by directly observing eating behaviors with a standardized protocol in a sample of10 children and adolescents with ASD residing at our rehabilitation institution comparing to 10 children and adolescents affected by mild-moderate mental retardation residing in the same institution. Carefully detailed food records for a 20 day period were analyzed.Subjects with ASD resulted significantly more selective than controls (lunch p = 0.016, dinner p = 0.042). Furthermore, the observed subjects with ASD had a bimodal distribution of weight percentiles ( 6 kids were < 25 % weight percentile. 2 kids > 75% weight percentile) while the control group revealed normal body weight distribution. We found a negative correlation between: food selectivity and duration of stay ( R = - 0.5848 ), as well as food selectivity and age ( R = - 0.6437 ), but a positive correlation between food refusal and disease severity measured with Autism Diagnostic Observation Scale (ADOS- 2) ( R = 0.4441 ). Our data confirm the feasibility of a direct observation monitoring protocol for feeding behavior importance of food selectivity in subjects affected by ASD. Younger children are more selective than older ones and the duration of institutional residency seems to positively impact this behavior pattern. Lastly, the severity of ASD symptoms resulted directly proportional to nutritional fixation severity

Titolo: SPSd - A New Rating Scale for Psychological Distress in Adolescence: a Validation Study on a Nationwide Italian Cohort Of 31,661 Adolescents (2016)

Autore: Enzo Grossi, Gabriele Manella, Giovanni Pieretti, Bruno Genetti, Milena Sperotto, Paolo Vian

Info: Psychological Distress, Nova Science Publishers Inc 2016



In this paper we present a new rating scale for adolescent psychological distress based on an Italian population study carried out in a cohort of 31,661 adolescents, and analyzed with Rasch Analysis in addition to the traditional tests. The Italian epidemiological survey on drug use in the school population was conducted by the Department of Anti Drug Policies in the first six months of 2014. It was based on a methodology of the European Monitoring Centre for Drugs and the Drug Addiction (EMCDDA - European School Survey Project on Alcohol and Other Drugs - ESPAD). The questionnaire and the survey procedures were taken from ESPAD and adjusted modified in order to address the specific characteristics of to the Italian school system. The total population sample consisted on 31,661 15-19 years old students who attended Italian public high schools. Among the items in the source scale we have selected nine items belonging to three dominions (Energy, Emotional Stability, Impulsivity and Risk-Taking) from which we expected the maximal indirect relation to wellbeing state and therefore as complement, to distress. To evaluate the psychometric properties of distress scale we carried out the homogeneity analysis or internal consistency of the scale, using the Cronbach's alpha coefficient, and analysis of the validity of the scale, using the Gamma coefficient of Goodman and Kruskal - index rank correlation. The data dimensionality was explored using Rasch Analysis and factor analysis. The Cronbach's alpha obtained considering all items was to 0.694, making it quite satisfactory and meeting the standards criteria of acceptability. Rasch analysis confirmed the items validity and that the items distribution remains in the area of the tolerance defined. A very good distribution emerges with a complete absorption of possible values along the scale range; neither ceiling or floor effect is observed. Whereas the level of distress measured by the scale is higher the lower the score detected, it is observed a significantly higher level of distress in females than in males (52.59 vs 59.61). For all psychotropic substances analyzed,significantly higher levels of distress in consumers than non-consumers have been found. In addition,the level of distress as measured by the scale appears to increase going from users of “soft” drugs to users of “hard” drugs: - 57.18 is the mean value obtained in the distress scale from the group of students who say they have never used any illegal drug; - 53.00 is the mean value obtained in the distress scale from the group of students who report use of cannabis at least once in their life; - 50.18 is the mean value obtained in the distress scale from the group of students who report use of cocaine at least once in their life; - 47.95 is the mean value obtained in the distress scale from the group of students who report use of heroin at least once in their life. This survey has detected several factors/conditions associated to the use of substances, which can be considered a reliable proxy of psychological distress: family background seems to be very important as well as the relationship with parents; this emerged as regards the presence of rules on behavior outside the home, the quality of relationships with prominent adult figures (particularly the father), their leadership role, and the family harmony.

Titolo: Psychological Interventions to Reduce Distress and Improve Quality of Life in Families with Autistic Children (2016)

Autore: Cristina Zarbo, Angelo Compare, Sara Melli, Enzo Grossi

Info: Psychological Distress, Nova Science Publishers Inc 2016



The experience of childbirth and becoming a parent can become particularly distressing when newborn has a severe chronic disability, like Autism Spectrum Disorder (ASD). Families with autistic children face an overload of their adaptive capacities and crisis situations that can in turn lead to various negative psychological consequences on each parent, the couple, children, and the whole family. Indeed, parents of autistic children show high incidence of psychological distress and mental disorders (e.g., anxiety disorders, depressive symptoms). Evidence suggests that parents of children with ASD may benefit of comprehensive and flexible psychological interventions to reduce parental distress and promote the wellbeing and improve the quality of life of the family.

Titolo: Mental Stress in Parents and Siblings of Autistic Children: Review of the Literature and Original Study of the Related Psychological Dimensions (2016)

Autore: Sara Melli, Enzo Grossi, Cristina Zarbo, Angelo Compare

Info: Psychological Distress, Nova Science Publishers Inc 2016



Parental mental stress is clinically common in families of autistic children and adversely affects the care of the child. Moreover, parents of autistic children frequently experience feelings of guilt, maladaptive coping styles, a lack of ability to forgive themselves and variations in mindfulness. However,it is unclear which of these dimensions is predominant in these families and their pattern of association with other components of this complex picture. While most notable in parents, this is also true of healthy siblings who intrinsically perceive more responsibility and often receive less attention than their afflicted brothers or sisters. The evidence available in the literature on the topic is quite controversial, revealing both positive and negative effects deriving from growing up with ASD siblings. Demographic and psychological information on mental stress, feelings of guilt, ability to forgive, mindfulness and coping styles were collected through clinical interviews and self-report questionnaires in parents and siblings of autistic children. Artificial Neural Networks (Auto-CM system) were applied to parents data to highlight the associations among the variables under investigation. Predominant dimensions in parents of autistic children indcluded low feelings of guilt, high levels of forgiveness and low levels of maladaptive coping responses. These three main dimensions were strictly related among themselves. While high parental mental stress was strictly related to high parental distress subscales, to high maladaptive coping styles, and to low self-forgiveness ability, conversely, low mental stress appeared to be marginal in relation to the other psychological dimensions. This behavior is typical of complex nonlinear systems. The severity of the ASD was not related to parental psychological dimensions. The ADOS scores, both low and high, were in fact marginal in the connectivity map in relation to the other dimensions. In siblings, the average distress scores level was extremely high irrespectively of ASD severity or family size. About one third of them were at risk of developing major depression. Distress appeared to be mainly related to difficulties in building a meaningful relationship with the ASD child and in managing the child’s behavioral problems. In conclusione the interplay of psychological factors related to stress in parents and siblings of ASD children is complex. Understanding these relationships is the starting point to activating and enhancing parental resources essential to the wellbeing of both children and caregivers.

Titolo: Artificial Neural Networks Link One-Carbon Metabolism to Gene-Promoter Methylation in Alzheimer's Disease (2016)

Autore: Enzo Grossi, Andrea Stoccoro, Pierpaola Tannorella, Lucia Migliore, Fabio Coppedè

Info: Journal Alzheimer Disease Jul 1;53(4):1517-22. doi: 10.3233/JAD-160210.



Background: There is increasing interest in DNA methylation studies in Alzheimer’s disease (AD), but little is still known concerning the relationship between gene-promoter methylation and circulating biomarkers of one-carbon metabolism in patients. Objective: To detect the connections among circulating folate, homocysteine (hcy) and vitamin B12 levels and promoter methylation levels of PSEN1, BACE1, DNMT1, DNMT3A, DNMT3B, and MTHFR genes in blood DNA. Methods:We applied a data mining system called Auto Contractive Map to an existing database of 100 AD and 100 control individuals. Results: Low vitamin B12 was linked to the AD condition, to low folates, and to high hcy. Low PSEN1 methylation was linked to low folate levels as well as to low promoter methylation of BACE1 and DNMTs genes. Low hcy was linked to controls, to high folates and vitamin B12, as well as to high methylation levels of most of the studied genes Conclusions: The present pilot study suggests that promoter methylation levels of the studied genes are linked to circulating levels of folates, hcy, and vitamin B12.

Titolo: Gut to brain interaction in Autism Spectrum Disorders. Role of probiotics on clinical, biochemical and neurophysiological parameters: a randomized controlled trial. (2016)

Autore: Elisa Santocchi, Letizia Guiducci, Francesca Fulceri, Lucia Billeci, Emma Buzzigoli, Fabio Apicella, Sara Calderoni, Enzo Grossi, Maria Aurora Morales, Filippo Muratori.

Info: BMC Psychiatry 16:183



Background A high prevalence of a variety of gastrointestinal (GI) symptoms is frequently reported in patients with Autism Spectrum Disorders (ASD). The GI disturbances in ASD might be linked to gut dysbiosis representing the observable phenotype of a “gut-brain axis” disruption. The exploitation of strategies which can restore normal gut microbiota and reduce the gut production and absorption of toxins, such as probiotics addition/supplementation in a diet, may represent a non-pharmacological option in the treatment of GI disturbances in ASD. The aim of this randomized controlled trial is to determine the effects of supplementation with a probiotic mixture (Vivomixx®) in ASD children not only on specific GI symptoms, but also on the core deficits of the disorder, on cognitive and language development, and on brain function and connectivity. An ancillary aim is to evaluate possible effects of probiotic supplementation on urinary concentrations of phthalates (chemical pollutants) which have been previously linked to ASD. Methods A group of 100 preschoolers with ASD will be classified as belonging to a GI group or to a Non-GI (NGI) group on the basis of a symptom severity index specific to GI disorders. In order to obtain four arms, subjects belonging to the two groups (GI and NGI) will be blind randomized 1:1 to regular diet with probiotics or with placebo for 6 months. All participants will be assessed at baseline, after three months and after six months from baseline in order to evaluate the possible changes in: GI symptoms; autism symptoms severity; affective and behavioral comorbid symptoms; plasmatic, urinary and fecal biomarkers related to abnormal intestinal function; neurophysiological patterns. Discussion The effects of treatments with probiotics on children with ASD need to be evaluated through rigorous controlled trials. Examining the impact of probiotics not only on clinical but also on neurophysiological patterns, the current trial sets out to provide new insights into the gut-brain connection in ASD patients. Moreover, results could add information to the relationship between phthalates levels, clinical features and neurophysiological patterns in ASD. Trial Registration: Identifier NCT02708901. Registered 4 March 2016. Key words Autism Spectrum Disorders (ASD), Gut-brain axis, endophenotype, probiotic Vivomixx®, quantitative electroencephalography (QEEG), phtalates.

Titolo: Microbiota Network and Mathematic Microbe Relation in Colostrum and Mature Milk of two different geographic areas: Italy versus Burundi (2016)

Autore: Lorenzo Drago, Marco Toscano, Roberta De Grandi, Enzo Grossi, Ezio Maria Padovani, Diego Peroni

Info: ISME Journal advance online publication 16 December 2016;



Human milk is essential for the initial development of newborns, as it provides all nutrients and vitamins, such as vitamin D, and it represents a great source of commensal bacteria. Here we explore the microbiota network of colostrum and mature milk of Italian and Burundian mothers. In both populations, by studying colostrum and mature milk microbiota with a new methodology based on an Artificial Neural Network (ANN) architecture, the Auto Contractive Map (AutoCM), it has been showed a different bacterial network, as diverse microbial hubs and central hubs have been observed in these samples. Interestingly, the Italian and Burundian mature milk showed a greater abundance of anaerobe intestinal bacteria if compared with colostrum samples, corroborating the hypothesis that intestinal microorganisms are able to reach the mammary glands by mean of dendritic cells and CD18+. In conclusion, the microbiota of human milk is a dynamic and complex ecosystem that not only differs between different populations, but it changes also during the transition from colostrum to mature milk. Finally, the association of complex mathematic systems such as ANN and AutoCM to metagenomics analysis represents an innovative approach to investigate in detail specific bacterial interactions in biological samples.

Titolo: Unexpected improvement in core autism symptoms after long term treatment with probiotics (2016)

Autore: Enzo Grossi, Delia Dunca, Sara Melli & Vittorio Terruzzi

Info: SAGE Open Medical Case Reports Volume 4: 1– 5



Background Many studies report the existence of so-called gut-brain axis: a physiological framework in which the gut microbiota communicates with the CNS and vice-versa through neural, endocrine and immune pathways. If this is true, then it is plausible to expect that the modulation gut microbiota may be a tractable strategy in developing novel therapeutics for complex CNS disorders. Autism is definitely one of these disorders. Objectives To report the case of a 12 year old boy with ASD and severe cognitive disability in which an unexpected improvement of autistic core symptoms was observed after four months of probiotic treatment. Method The case study refers to a 12 year old boy with ASD, severe cognitive disability and gastrointestinal disturbances attending the Villa Santa Maria Institute in resident care since 2009. The boy was first diagnosed with Pervasive Developmental Disorder (PDD) at the age of two. Later on ASD Diagnosis was made according to DSM-V criteria and was confirmed by the ADOS-2 assessment (Autism Diagnostic Observation Schedule). In February 2014, a probiotic treatment was prescribed to control gastrointestinal symptoms (VSL#3, a product with lyophilized Bifidobacteria, Lactobacilli and Streptococci), which lasted four months. The diet was consistently followed throughout the entire period and treatment compliance was ensured by a careful supervision. No other dietary supplements containing probiotics or prebiotics or antibiotics was administered during the treatment and follow-up period. The rehabilitation program already in place, based on behavior, communication, psychomotor therapy, individualized Education Plan/School Services and parents involvement, was regularly maintained. Improvement in ASD symptoms already became apparent and clinically evident from the few weeks after starting the probiotic treatment. The ADOS-2 assessment was repeated six times over the October 2013-March 2015 period (twice before starting the treatment, twice during (after month 1 and month 4) and twice after treatment interruption. Results The probiotic treatment reduced the severity of abdominal symptoms as expected. While the Restricted Repetitive Behaviors domain scores did not change, an unexpected improvement of the Autistic core symptoms did occur. The Social Affect Domain Score of ADOS improved, going from 20 to 18 after two months of treatment, with a further 1 point reduction during the following two months. The severity level 17 remained stable during the follow up period. Conclusions The appropriate use of probiotics merits further research, which hopefully will open new avenues in the fight against ASD.

Titolo: Predictors of Response to Long-Term Cholinesterase Inhibitors Treatment of Alzheimer Disease: Data Mining from Tredem Registry (2016)

Autore: Maurizio Gallucci , Pierpaolo Spagnolo, Maria Aricò, Enzo Grossi

Info: Journal Alzheimer Disease Vol. 50 969-979


Abstract. Background: the pharmacological treatment of Alzheimer Disease is based largely on cholinesterase inhibitors. Objective: to investigate whether or not some non-pharmacological and contextual factors measured prior to starting treatment such as past occupation, lifestyles, marital status, degree of autonomy and cognitive impairment, living alone or with others, the degree of brain atrophy are associated with a better response to cholinesterase inhibitors treatment. Methods: Eighty-four Alzheimer’s disease (AD) and six AD with cerebrovascular disease (AD + CVD) outpatients of Treviso Dementia (TREDEM) Registry, with an average cholinesterase inhibitors treatment length of four years, were considered. The outpatients had undergone a complete evaluation and some non-pharmacological and contextual factors were collected. We defined responder a patient with a delta score T0 – T1 equal or inferior to 2.0 points per year of MMSE and a non-responder a patient with a delta score T0 – T1 superior to 2.0 points per year. In order to identify hidden relationships between variables related to response and non-response, we use a special kind of artificial neural network called Auto-CM, able to create a semantic connectivity map of the variables considered in the study. Results: a higher cognitive profile, a previous intellectual work, healthier lifestyles, being married and not living alone, a higher degree of autonomy and lower degree of brain atrophy at baseline resulted to affect the response to long term ChEI therapy. Conclusion: non-pharmacological and contextual factors appear influence the effectiveness of treatment with ChEI in the long term.

Titolo: Pregnancy risk factors in autism: a pilot study with artificial neural networks (2016)

Autore: Enzo Grossi, Federica Veggo, Antonio Narzisi, Angelo Compare, Filippo Muratori

Info: Pediatric Research; Pediatr Res. 2016 Feb;79(2):339-47. doi: 10.1038/pr.2015.222. PubMed PMID: 26524714.


Background: Autism is a multi-factorial condition where a single risk factor can unlikely provide comprehensive explanation for the disease origin. Moreover, due to the complexity of risk factors interplay, traditional statistics is often unable to explain the core of the problem due to the strong inherent non-linearity of relationships. The aim of this study was to assess the frequency of twenty-seven potential risk factors related to pregnancy and peri-post natal period. Methods: The mothers of of forty-five autistic children and of 68 typical developing children completed a careful interview. 24 siblings of 19 autistic children formed an internal control group. Results: A higher prevalence of potential risk factors was observed in twenty-two and fifteen factors in external control and internal control group respectively. For six of them, the difference in prevalence was statistically significant. Specialized Artificial Neural Networks (ANNs) discriminated between autism and control subjects with 80.19% global accuracy. Conclusions: Pregnancy factors play an important role in autism development. ANNs are able to build up a predictive model, which could represent the basis for a diagnostic screening tool.

Titolo: Data Mining of Determinants of Intrauterine Growth Retardation Revisited Using Novel Algorithms Generating Semantic Maps and Prototypical Discriminating Variable Profiles (2015)

Autore: Massimo Buscema, Enzo Grossi, Luisa Montanini, Maria E. Street

Info: Plos One 7th July 2015



Objectives: Intra-uterine growth retardation is often of unknown origin, and is of great interest as a “FetalOrigin of Adult Disease” has been now well recognized. We built a benchmark based upon a previously analysed data set related to Intrauterine Growth Retardation with 46 subjectsdescribed by 14 variables, related with the insulin-like growth factor system and pro-inflammatorycytokines, namely interleukin -6 and tumor necrosis factor -α. Design and Methods: We used new algorithms for optimal information sorting based on the combination of twoneural network algorithms: Auto-contractive Map and Activation and Competition System. Auto-Contractive Map spatializes the relationships among variables or records by constructinga suitable embedding space where ‘closeness’ among variables or records reflects accurately their associations. The Activation and Competition System algorithm insteadworks as a dynamic non linear associative memory on the weight matrices of other algorithms, and is able to produce a prototypical variable profile of a given target. Results: Classical statistical analysis, proved to be unable to distinguish intrauterine growth retardationfrom appropriate-for-gestational age (AGA) subjects due to the high non-linearity of underlying functions. Auto-contractive map succeeded in clustering and differentiatingcompletely the conditions under study, while Activation and Competition System allowed to develop the profile of variables which discriminated the two conditions under study betterthan any other previous form of attempt. In particular, Activation and Competition System showed that ppropriateness for gestational age was explained by IGF-2 relative gene expression, and by IGFBP-2 and TNF-α placental contents. IUGR instead was explained by IGF-I, IGFBP-1, IGFBP-2 and IL-6 gene expression in placenta. Conclusions: This further analysis provided further insight into the placental key-players of fetal growthwithin the insulin-like growth factor and cytokine systems. Our previous published analysis could identify only which variables were predictive of fetal growth in general, and identified only some relationships.

Titolo: Networks in Coronary Heart Disease Genetics as a Step towards Systems Epidemiology. (2015)

Autore: Fotios Drenos, Enzo Grossi, Massimo Buscema, Steve E Humphries

Info: Plos One May 7, 2015



We present the use of innovative machine learning techniques in the understanding of Coronary Heart Disease (CHD) through intermediate traits, as an example of the use of this class of methods as a first step towards a systems epidemiology approach of complex diseases genetics. Using a sample of 252 middle-aged men, of which 102 had a CHD event in 10 years follow-up, we applied machine learning algorithms for the selection of CHD intermediate phenotypes, established markers, risk factors, and their previously associated genetic polymorphisms, and constructed a map of relationships between the selected variables. Of the 52 variables considered, 42 were retained after selection of the most informative variables for CHD. The constructed map suggests that most selected variables were related to CHD in a context dependent manner while only a small number of variables were related to a specific outcome. We also observed that loss of complexity in the network was linked to a future CHD event. We propose that novel, non-linear, and integrative epidemiological approaches are required to combine all available information, in order to truly translate the new advances in medical sciences to gains in preventive measures and patients care.

Titolo: Age-, Adiposity- and Insulin-Related Changes in Urinary Di-(2-Ethylhexyl) Phthalate Metabolites (2015)

Autore: Arianna Smerieri, Chiara Testa, Pietro Lazzeroni, Francesca Nuti, Enzo Grossi, Silvia Cesari, Luisa Montanini, Giuseppe Latini, Sergio Bernasconi, Anna Maria Papini, Maria E. Street.

Info: Plos One 10: 2. 02



Background and objectives: Phthalates might be implicated with obesity and insulin sensitivity. The aim of this study was to evaluate the levels of primary and secondary metabolites of DEHP in urine in obese and normal-weight subjects both before and during puberty, and to investigate their relationships with auxological parameters and indexes of insulin sensitivity. Methods: DEHP metabolites (MEHP, 6-OH-MEHP, 5-oxo-MEHP, 5-OH-MEHP, and 5-CX-MEHP), were measured in urine by RP-HPLC-ESI-MS. Traditional statistical analysis and a data mining analysis through the Auto-CM system, a fourth generation artificial neural network (ANN), were able to offer an insight of the complex biological connections between the studied variables. Results: The data showed changes in DEHP metabolites in urine, related with obesity, onset of puberty, and presence of insulin resistance. Changes in urine metabolites were related with age, height and weight, waist circumference and waist to height ratio, thus to fat distribution. In addition, clear relationships in both obese and normal-weight subjects were detected among MEHP, its products of oxidation and measurements of insulin sensitivity (insulin, fasting glucose to insulin ratio, HOMA-IR index, and whole body insulin sensitivity index). Conclusions: It remains to be elucidated whether exposure to phthalates per se is actually the risk factor or if the ability of the body to metabolize phthalates is actually the key point. Further studies that span from conception to elderly subjects besides further understanding of DEHP metabolism are warranted to clarify these aspects.

Titolo: Which are the better outcome predictors in preschoolers with autism who respond positively at the treatment as usual (TAU) ? Insights from an observational study using Artificial Neural Networks (2015)

Autore: Antonio Narzisi, Filippo Muratori, Massimo Buscema, Sara Calderoni, Enzo Grossi IRCCS Stella Maris Foundation - Department of Developmental Neuroscience, University of Pisa, Pisa, Italy, Semeion Research Center, Rome, Italy, 4Department of Mathematical



In Italy Treatment As Usual (TAU) for autism spectrum disorders (ASD) is composed of specific treatments performed by child neuropsychiatric services and of school inclusion with individual support teacher. The Artificial Neural Networks (ANNs) have never been used in order to study the effects of treatment in ASD. Auto Contractive Map (Auto-CM) is a special kind of ANNs able to find out consistent trends and associations among variables creating a semantic connectivity map. The matrix of connections, visualized through minimum spanning tree filter takes into account non linear associations among variables and captures connection schemes among clusters. Objectives: The main aim is to use Auto-CM in order to identify key variables to discriminate among responders versus no responders at TAU. Methods: 56 preschoolers with ASD aged between 24 and 48 months were recruited at different centers in Italy. They were evaluated by blind researchers at baseline and after six months using a multi-informant assessment protocol. All children were referred to community providers for available interventions. Results: At T1, the severity of autism measured through ADOS improved in 62% of involved children (Response) while it was the same or worse in 37% of 56 (No Response). The application of the the Semeion ANNs overcome the 85% of global accuracy (Sine–Net almost reaching 90%). Consequently, some of the tested algorithms were able to find a good correlation between some variables and TAU outcome. The semantic connectivity map obtained with the application of the Auto-CM system showed results that clearly indicated that ‘Response’ can be visually separated from the ‘No Response’ cases. Particularly it was possible to visualize a Response area characterized by’ Parents Involvement high’ and ‘MacArthur Expressive low’. No Response area resulted strongly connected only with ‘Parents Involvement low’. Conclusions: The ANN model that was used in this study appears to be a promising tool for the identification of the variables involved in the positive of negative response to TAU in autism.

Titolo: L'apporto dell'arte, della religione e della comunicazione nella "cura" delle persone con disturbi dello spettro autistico (2014)

Autore: Marina Norsi

Info: Dolentium Hominum n. 86



L'apporto della religione e dell'arte nella cura dei bambini autistici viene analizzata basandosi sui dati riportati nella letteratura e sull'esperienza degli ultimi 10 anni nelle strutture terapeutiche(nidi e asili) in Israele. L'influenza della religione nell'accettare la diagnosi ed affrontare la cura dell'autismo puo' essere positiva(accettazione della malattia come espressione della volonta' Divina) o negativa(malattia come punizione divina per i dubbi dei genitori nei confronti di Dio) . I dati riportati in letteratura confermano che il supporto delle organizzazioni religiose e dei ministri del culto sono di grande aiuto: diminuiscono lo stress e lo stato di ansia delle famiglie. L'arte terapia usata ormai in tutto il mondo come terapia complementare ha vantaggi rispetto alle terapie convenzionali:e' un mezzo di comunicazione non verbale, viene accettata dai bambini in modo positivo e non minaccioso,potenzia il contatto di sguardo, potenzia l'apprendimento di colori, forme geometriche, oggetti. Sono stati descritti in modo particolareggiato esempi concreti dell'uso dell'arte terapia e pratiche religiose nella stesura dei programmi cognitivi, educativi nelle strutture terapeutiche per bambini autistici.

Titolo: The Role of Intestinal Dysbiosis in the Pathogenesis of Autism: Minireview (2014)

Autore: Enzo Grossi & Vittorio Terruzzi

Info: International Journal of Microbiology & Advanced Immunology (IJMAI) 2: 201



Autism spectrum disorder is a complex neurodevelopmental disease where gastrointestinal disturbance is commonly reported. Here we review the evidence suggesting that gut microbota my play a role in this disease and summarize comparative studies we found in international literature on the topic. Discussion of results, methodology of the data collection, bias of selection and behavioral interferences lead to the conclusion that changes in the gut microbiota is a significant piece of autism spectrum disorder but further studies are needed to understand this pathogenetic role.

Titolo: Personality traits, cardiac risk factors, and their association with presence and severity of coronary artery plaque in people with no history of cardiovascular disease. (2014)

Autore: Angelo Compare, Paula M C Mommersteeg, Francesco Faletra, Enzo Grossi, Elena Pasotti, Tiziano Moccetti, Angelo Auricchio

pdf url


Coronary artery disease (CAD) is a complex, multifactorial disease in which personality seems to play a role but with no definition in combination with other risk factors. Objective: To explore the nonlinear and simultaneous pathways between traditional and personality traits risk factors and coronary stenosis by Artificial Neural Networks (ANN) data mining analysis. Method: Seventy-five subjectswere examined for traditional cardiac risk factors and personality traits.Analyseswere based on a new data mining method using a particular artificial adaptive system, the autocontractive map (AutoCM). Results: Several traditional Cardiovascular Risk Factors (CRF) present significant relations with coronary artery plaque (CAP) presence or severity. Moreover, anger turns out to be the main factor of personality for CAP in connection with numbers of traditional risk factors. Hidden connection map showed that anger, hostility, and the Type D personality subscale social inhibition are the core factors related to the traditional cardiovascular risk factors (CRF) specifically by hypertension. Discussion: This study shows a nonlinear and simultaneous pathway between traditional risk factors and personality traits associated with coronary stenosis in CAD patients without history of cardiovascular disease. In particular, anger seems to be the main personality factor for CAP in addition to traditional risk factors.

Titolo: PAHA study: Psychological Active and Healthy Aging: psychological wellbeing, proactive attitude and happiness effects of whole-body vibration versus Multicomponent Training in aged women: study protocol for a randomized controlled trial. (2014)

Autore: Angelo Compare, Cristina Zarbo, Elena Marín, Alessia Meloni, Jacobo A Rubio-Arias, Rosendo Berengüí, Enzo Grossi, Edo Shonin, Gianmaria Martini, Pedro E Alcaraz



Evidence demonstrates that physical exercise and psychological wellbeing are closely interlinked, particularly in older-aged women. However, research investigating how different forms of exercise influence mental health in older-aged women is underdeveloped. Methods/design: A randomized controlled trial (N = 300) will assess the relative effectiveness of two different exercise programs (whole-body vibration and Multicomponent Training) for improving psychological wellbeing in older-aged women. The following outcomes will be assessed at three time points (that is, pre, post, and follow-up): psychological wellbeing, proactive attitude, quality of life, and happiness.

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