DEPARTMENT OF NEUROSCIENCE

STAFF

Head Gianluigi FORLONI, Biol.Sci.D. Laboratory of Biology of Neurodegenerative Disorders

Head Gianluigi FORLONI, Biol.Sci.D.

Genetic of Neurodegenerative disorders Unit Head Diego ALBANI, Biol Sci. D. Acute Spinal trauma and regeneration Unit Head Pietro VEGLIANESE, Ph D Laboratory of Cell Death and Neuroprotection Head Tiziana BORSELLO, Biol.Sci.D. Laboratory of Epidemiology and Social Psychiatry Head Barbara D’AVANZO, Philos.D. Laboratory of Experimental Neurology

Head Annamaria VEZZANI, Biol.Sci.D. Physopathology of glia-neuron communication Unit Head Teresa RAVIZZA, Biol. Sci.D Laboratory of Experimental Psychopharmacology Head Luigi CERVO, Ph.D. Laboratory of Geriatric Neuropsychiatry

Head Ugo LUCCA, MSc Geriatric Epidemiology Unit Head Mauro TETTAMANTI, Biol.Sci.D. Geriatric Pharmacology Unit Head Emma RIVA, M.D. Laboratory of Inflammation and Nervous System Diseases

Head Maria Grazia DE SIMONI, Biol.Sci.D Cell therapy and Acute Brain Injury Unit Head Elisa RONCATI ZANIER. M D

Laboratory of Molecular Neurobiology

Head Caterina BENDOTTI, Pharm.D. Laboratory of Neurochemistry and Behavior Head Roberto William INVERNIZZI, Biol. Sci D

Pharmacology of Cognitive Behavior Unit

Head Mirjana CARLI, Ph.D. Laboratory of Neurological Disorders

Head Ettore BEGHI, M.D. Laboratory of Prion Neurobiology Head Roberto CHIESA, Biol. Sci. D Laboratory of Quality Assessment of Geriatric Services Unit Head Alessandro NOBILI, M.D. Pharmacotherapy and Presciption Appropriateness Head Luca PASINA, Phar. Dr

CURRICULA VITAE

Gianluigi Forloni, obtained the Degree of Biological Science at the University of Milan in 1985. After two years of post doc at the Department of Neuroscience and Psychiatry at Johns Hopkins University in Baltimore, USA, he came back to the Mario Negri Institute and between 1992 and 1996 he was the head of the Neurobiology of Alzheimer's disease Unit; since 1996 he is the Head of the Biology of Neurodegenerative Diseases Lab and since 2002 the Head of the Neuroscience Department. His scientific interest is focused on the biological and genetic bases of aging-related disorders in particular Alzheimer’s disease, Prion-related encephalopathies and Parkinson’s disease. He has been member of several European committees for the examination of projects in the neuroscience field. He is now member of the Coordination group of the European IMI Consortium PharmaCog. He is President of the Italian Association on Brain Aging Research (AIRIC), member of the Scientific Committee of the Dementia section of the Italian Society of Neurology (SINDEM) and member of the European Academy of Sciences. He is the author of more than 270 peer-reviewed scientific articles and about 30 reviews or book chapters. Selected publications

Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O.,Tagliavini F. Neurotoxicity of a prion protein fragment. Nature 362: 543-546 (1993)

Forloni, G., Tagliavini, F.,Bugiani, O. and Salmona, M. Amyloid in Alzheimer’s disease and prion-related encephalopathies: Studies with synthetic peptides. Progr. Neurobiol. 49: 287- 315 (1996)

Forloni G. Iussich, S. Awan T. Colombo L. Angeretti, N. Girola, L. Bertani, I. Poli, G. Caramelli, M. Bruzzone, MG.Farina, L. Limido, L. Rossi, G. Giaccone G. Ironside, JW. Bugiani, O.Salmona M. and Tagliavini, F. Tetracyclines affect prion infectivity Proc. Natl. Acad. Sci . New York 99: 10849-10854 (2002)

Fioriti, L. Angeretti, N.. Colombo, L., De Luigi A., Manzoni, C., Colombo A., Morbin, M., Tagliavini, F., Salmona, M. Chiesa, R. Forloni, G. Neurotoxic and gliotrophic activity of a synthetic peptide homologous to Gerstmann-Sträussler-Scheinker disease amyloid protein. J. Neurosci. 27: 576-83 (2007)

Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron. 60: 598-609 (2008)

Balducci, C., Beeg, M., Stravalaci, M., Bastone, A.,, Sclip, A., Biasini, E., Tapelll., Colombo, L. Canzoni, C., Borsello, T., Chiesa, R., Gobbi, M., Salmona M. Forloni, G., A oligomers impair memory independently of cellular prion potei Proc. Natl. Acad. Sci USA, 107: 2295-2300 (2010)

Puoti, G., Bizzi, A., Forloni G., Safar JG.,Tagliavini, F., Gambetti, P. Sporadic human prion diseases: molecular insights and diagnosis. Lancet Neurology 11: 618-28 (2012)

Polito, l, Kehoe P, Davin, A., Ghidoni, R., Benussi, L., Quadri, PL, Lucca, U., Tettamanti, M., Mariani, C., Forloni, G., Albani, D. The SIRT2 polymorphism rs10410544 as susceptibility factor for Alzheimer’s disease. Alzheimer & Dementia 9: 392-399 (2013)

Balducci C., Minniti, S., Lavitola, P., Zotti, M., Sancini, G, Cagnotto, A, Salmona M, Haaparanta-Solin M, Forloni G, Masserini M, Re F. Efficacy of a nanomedicine-based therapeutics in Alzheimer's mouse model J Neurosci 34:14022-14031 (2014)

Haïk, S, Marcon, Coudert M, Tettamanti M, Welaratne A, Giaccone G, Azimi S, Pietrini V, Fabreguettes, J-R, Imperiale, D, Cesaro, P, Buffa, C, Aucan, C, Lucca, U, Mallet, A, Suardi, S, Tranchant, C, Zerr, I, Houillier, C, Redaelli, V, Vespignani, H, Campanella A, Sellal, F; Gobbi, M, Seilhean, D, Canovii, M, Sedel, F, Di Fede, G, Laplanche, JL, Pocchiari, M, Salmona, M, Forloni, G, Brandel, J-P Tagliavini, F. Doxycycline in Creutzfeldt-Jakob disease: a phase 2, randomized, double-blind, placebo-controlled trial Lancet Neurol. 13:150-8 (2014)

Forloni G, Tettamanti M, Lucca U, Albanese Y, Quaglio E, Chiesa R,Villani F, Erbetta E, Redaelli V, Tagliavini F, Artuso V, Roiter I. Preventive study in subjects at risk of FFI: innovative approach to rare diseases. Prion 9: 75-9 (2015)

Micotti E, Paladini A, Balducci C, Tolomeo D, Frasca A, Marizzoni M, Filibian M, Caroli A, Valbusa G, Dix S, O'Neill, M, Ozmen L, Czech C, Richardson CJ, Frisoni GB, Forloni G Striatum and entorhinal cortex common neuropathological targets in mouse models of Alzheimer's disease. Neurobiology of Aging 76: 776-788 (2015)

Ettore Beghi graduated in Medicine in 1972 and received his specialty in neurology in 1976 at the University of Milan. He trained in epidemiology with a fellowship at the Department of statistics and Epidemiology of the Mayo Clinic in Rochester, MN (USA). He is Head of the Laboratory of Neurological Disorders at the Mario Negri Institute, Director of the Neurophysiology/Epilepsy Unit and Professor of Neuroepidemiology at the University of Milano-Bicocca, Monza. He is member of the editorial board of the journals Epilepsia, Neuroepidemiology, Inpharma, Drugs in R & D, Clinical Drug Investigation, Neurological Sciences and is a referee of several national and international medical journals. The main areas of interest and research include studies on the descriptive, analytic, and experimental epidemiology in the field of epilepsy, peripheral neuropathies, headache, and amyotrophic lateral sclerosis. Selected publications

Beghi E, D'Alessandro R, Beretta S, Consoli D, Crespi V, Delaj L, Gandolfo C, Greco G, La Neve A, Manfredi M, Mattana F, Musolino R, Provinciali L, Santangelo M, Specchio Lm, Zaccara G; On Behalf Of The Epistroke Group. Incidence And Predictors Of Acute Symptomatic Seizures After Stroke Neurology 2011; 77:1785-1793.

E. Beghi, E. Pupillo, P. Messina, G. Giussani, A. Chio, S. Zoccolella, C. Moglia, M. Corbo, G. Logroscino, For The Eurals Group. Coffee And Amyotrophic Lateral Sclerosis: A Possible Preventive Role. Am J. Epidemiol 2011; 174 : 1002-1008.

Beghi E, Pupillo E, Bonito V, Buzzi P, Caponnetto C, Chiò A, Corbo M, Giannini F, Inghilleri M, Bella Vl, Logroscino G, Lorusso L, Lunetta C, Mazzini L, Messina P, Mora G, Perini M, Quadrelli Ml, Silani V, Simone Il, Tremolizzo L; Italian Als Study Group. Randomized Double-Blind Placebo-Controlled Trial Of Acetyl-L-Carnitine For ALS. Amyotroph Lateral Scler Frontotemporal Degener. 2013:14:397-405.

Pupillo E, Messina P, Logroscino G, Beghi E; the SLALOM Group. Long-term survival of amyotrophic lateral sclerosis. A population-based study. Ann Neurol 2014; 75: 287-297.

Pupillo E, Messina P, Giussani G, Logroscino G, Zoccolella S, Chiò A, Calvo A, Corbo M, Lunetta C, Marin B, Mitchell D, Hardiman O, Rooney J, Stevic Z, Bandettini di Poggio M, Filosto M, Cotelli MS, Perini M, Riva N, Tremolizzo L, Vitelli E, Damiani D, BEGHI E; the EURALS Consortium. Physical Activity and ALS. A European Population-based, Case-control Study. Ann Neurol 2014; 75: 708-716.

Giussani G, Franchi C, Messina P, Nobili A, BEGHI E; the EPIRES Group. Prevalence and incidence of epilepsy in a well-defined population of Northern Italy. Epilepsia 2014; 55: 1526-1533.

Global Burden of Disease Study 2013 Collaborators* Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet 2015; 386(9995): 743-800.

Pupillo E, Bianchi E, Messina P, Chiveri L, Lunetta C, Corbo M, Filosto M, Lorusso L, Marin B, Mandrioli J, Riva N, Sasanelli F, Tremolizzo L, Beghi E and the EURALS Consortium. Extrapyramidal and cognitive signs in amyotrophic lateral sclerosis. Amyotroph Lateral Scler Frontotemporal Degener 2015; 16(5-6): 324-330.

GBD 2013 DALYs and HALE Collaborators. Global, regional, and national disability-adjusted life years (DALYs) for 306 diseases and injuries and healthy life expectancy (HALE) for 188 countries, 1990-2013: quantifying the epidemiological transition. Lancet 2015;386(10009):2145-2191.

Nobile-Orazio E, Cocito D, Jann S, Uncini A, Messina P, Antonini G, Fazio R, Gallia F, Schenone A, Francia A, Pareyson D, Santoro L, Tamburin S, Cavaletti G, Giannini F, Sabatelli M, BEGHI E; for the IMC Trial Group. Frequency and time to relapse after discontinuing 6-month therapy with IVIg or pulsed methylprednisolone in CIDP. J Neurol Neurosurg Psychiatry 2015; 86(7): 729-734.

Sarchielli P, Corbelli I, Messina P, Cupini LM, Bernardi G, Bono G, Di Piero V, Petolicchio B, Livrea P, Prudenzano MP, Pini LA, Sandrini G, Allena M, Tedeschi G, Russo A, Caproni S, Beghi E, Calabresi P; SAMOHA Study Group. Psychopathological comorbidities in medication-overuse headache: a multicentre clinical study. Eur J Neurol 2016;23:85-91.

Caterina Bendotti, got her degree in Pharmacy at the University of Milano in 1984; Research assistant in Neuropharmacology la of the Mario Negri Institute (IRFMN) until 1986. From 1986 to 1988 post doc at the Genetic developmental Lab, Dept. of Physiology of the Johns Hopkins University, Baltimore, USA. 1988 -1992 research fellow in the laboratory of Neuropharmacology and in the 1992, she became head of the Molecular Neurobiology Unit at the IRFMN and in 1998 become head of laboratory. The major research interest is the study of pathogenetic mechanisms of familial Amyotrophic Lateral Sclerosis.. Member of different scientific boards either editorial (J. Neurochemistry(2002-2012) and CNS & Neurological Disorders - Drug Targets from 2009), of international meetings (International Symposia on ALS held in Milano in 2003 and 2013 , ENCALS meetings in Leuven-Belgium in 2014). Member of the Italian Ministry of Health Committees for the diagnosis, cure, care and assistance of patients with ALS (2003-2007), member of the Board of Directors of the Italian Society of (2005-2009). Member of the Research Advisory Panel di MNDA UK (2006-2010), of Thierry Latran Foundation for the ALS, France from 2008 and of the Italian ALS Association (AISLA) from 2014. Co-organizer of the first international meeting on” Mutant SOD1 and familial ALS:from the molecule to man” held in Milano in 2007, of the workshop on MND at IBRO symposium in Firenze, Italy (2011), of the satellite symposium on MND of FENS in Milano (2014). Since 2011 is responsible of the Animal Facility of AriSLA. Scientific reviewer of 15 international scientific journals and 7 grant international agencies. First /Last Author and Co-Author of 143 peer-reviewed Articles ( H index= 37). Rapporteur of many communications in national and international meetings. Selected publications

De Paola M, Sestito SE, Mariani A, Memo C, Fanelli R, Freschi M, Bendotti C, Calabrese V, Peri F. Synthetic and natural small molecule

TLR4 antagonists inhibit motoneuron death in cultures from ALS mouse model. Pharmacol Res. 2015 103:180-187 Caron I, Micotti E, Paladini A, Merlino G, Plebani L, Forloni G, Modo M, Bendotti C. Comparative Magnetic Resonance Imaging and

Histopathological Correlates in Two SOD1 Transgenic Mouse Models of Amyotrophic Lateral Sclerosis. PLoS One. 2015 10(7):e0132159.

Violatto MB, Santangelo C, Capelli C, Frapolli R, Ferrari R, Sitia L, Tortarolo M, Talamini L, Previdi S, Moscatelli D, Salmona M, Introna M, Bendotti C, Bigini P. Longitudinal tracking of triple labeled umbilical cord derived mesenchymal stromal cells in a mouse model of Amyotrophic Lateral Sclerosis. Stem Cell Res. 2015 15(1):243-53.

Tortarolo M, Vallarola A, Lidonnici D, Battaglia E, Gensano F, Spaltro G, Fiordaliso F, Corbelli A, Garetto S, Martini E, Pasetto L,

Kallikourdis M, Bonetto V, Bendotti C. Lack of TNF-alpha receptor type 2 protects motor neurons in a cellular model of amyotrophic lateral sclerosis and in mutant SOD1 mice but does not affect disease progression. J Neurochem. 2015 135:109-24.

Lauranzano E, Pozzi S, Pasetto L, Stucchi R, Massignan T, Paolella K, Mombrini M, Nardo G, Lunetta C, Corbo M, Mora G, Bendotti C, Bonetto V. Peptidylprolyl isomerase A governs TARDBP function and assembly in heterogeneous nuclear ribonucleoprotein complexes. Brain. 2015 138(Pt 4):974-91.

Marino M, Papa S, Crippa V, Nardo G, Peviani M, Cheroni C, Trolese MC, Lauranzano E, Bonetto V, Poletti A, DeBiasi S, Ferraiuolo L, Shaw PJ, Bendotti C. Differences in protein quality control correlate with phenotype variability in 2 mouse models of familial amyotrophic lateral sclerosis. Neurobiol Aging. 2015, 36(1):492-504.

Acquadro E, Caron I, Tortarolo M, Bucci EM, Bendotti C, Corpillo D. Human SOD1-G93A specific distribution evidenced in murine brain of a transgenic model for amyotrophic lateral sclerosis by MALDI imaging mass spectrometry. J Proteome Res. 2014 4;13:1800-9

Peviani M, Tortarolo M, Battaglia E, Piva R, Bendotti C Specific induction of Akt3 in spinal cord motor neurons is neuroprotective in a mouse model of familial amyotrophic lateral sclerosis Mol Neurobiol 2014 49 : 136-148

Nardo G, Iennaco R, Fusi N, Heath PR, Marino M, Trolese MC, Ferraiuolo L, Lawrence N, Shaw PJ, Bendotti C. Transcriptomic indices of fast and slow disease progression in two mouse models of amyotrophic lateral sclerosis. Brain. 2013; 136:3305-32

Tiziana Borsello got her Degree in Biological Science at the University of Torino in 1990 and she then obtained a PhD in Neuroscience at the University of Turin Medical School. She won a 1 year fellowship from the European Science Foundation to work at the Netherlands Research Institute of Amsterdam. From 1997 to 1999 she was a Researcher at the Institute of Neurobiology, CNR, Rome Italy. In the period 1999-2003 she was Premier Assistant at the Département de Biologie Cellulaire et de Morphologie, Université de Lausanne, Switzerland, and then became Maitre Assistant and group leader in the same institute in 2004. In 2004 joined the Biol. Neurodeg. Disorders Lab at the "Mario Negri” Institute. In 2005 won the Prize of the Pfizer Foundation, Neuroscience and Diseases Nervous System. Since 2006 she is the Head of the Unit: Neuronal Death and Neuroprotection. Her main scientific interests focus on understanding the role of signalling pathways in neuronal death after different stress-stimuli and neuroprotection. In particular, the present research is focused on the study of the mechanisms leading to excitotoxic stress, ischemia, Traumatic Brain Injury and cell death pathways in neurodegenerative diseases such as Alzheimer, with the challenge to design more specific methods of neuroprotection.

Selected publications

Cimini S., Sclip A., Mancini S., Colombo L., Messa M., Cagnotto A., Di Fede G., Tagliavini F., Salmona M., and Borsello T. (2015) Neurobiol Dis. pii: S0969-9961(15)30115-7.

Vercelli A., Biggi S., Sclip A., Repetto I.E., Cimini S., Falleroni F., Tomasi S., Monti R., Tonna N., Morelli F., Grande V., Stravalaci M., Biasini E., Marin O., Bianco F., di Marino D. and Borsello T. The cell-permeable Aβ1-6A2VTAT(D) peptide reverts synaptopathy induced by Aβ1-42wt. Exploring the role of MKK7 in excitotoxicity and cerebral ischemia: a novel pharmacological strategy against brain injury (2015) Cell Death Dis.;6:e1854

Relja B., Weber R., Maraslioglu M., Wagner N., Borsello T., Jobin C., Marzi I and Lehnert M.Differential Relevance of NF-κB and JNK in the Pathophysiology of Hemorrhage/Resususcitation-Induced Liver Injury after Chronic Ethanol Feeding. (2015) Plos One. 10(9):e0137875.

Magini A., Polchi A., Tozzi A., Tancini B., Tantucci M., Urbanelli L., Borsello T., Calabresi P., Emiliani C. Abnormal cortical lysosomal β-hexosaminidase and β-galactosidase activity at post-synaptic sites during Alzheimer's disease progression. (2015) Int J Biochem Cell Biol. 58:62-70.

Tozzi A., Sclip A, Tantucci A., De Lure A., Griglieri V., Costa C., Di Filippo M., Borsello T, Calabresi P. Region- and age-dependent reductions of hippocampal long-term potentiation and NMDA to AMPA ratio in a genetic model of Alzheimer's diseaseNeurobiol Aging. 2015; 36:123-33

Davoli E., Sclip A, Cecchi M., Cimini S., Carrà A., Salmona M, Borsello T 2014 Determination of tissue levels of a neuroprotectant drug: the cell permeable JNK inhibitor peptide. J Pharmacol Toxicol Methods. 70: 55-61.

Sclip A, Tozzi A., Cardinetti D., Colombo I., Calabresi P., Salmona M, Welker E., Borsello T (2014) C-Jun N-Terminal Kinase Has A Key Role In Alzheimer Disease Synaptic Dysfunction In Vivo. Cell Death & Disease2014 Jan 23;5:e1019

Sclip A, Arnaboldi A, Colombo I, Veglianese P, Colombo L, Messa M, Mancini S, Cimini S, Morelli F, Antoniou X, Welker E, Salmona M, Borsello T. Soluble Aβ oligomer-induced synaptopathy: c-Jun N-terminal kinase's role. J Mol Cell Biol. 2013;5 277-9.

Fluharty BR, Biasini E, Stravalaci M, Sclip A, Diomede L, Balducci C, La Vitola P, Messa M, Colombo L, Forloni G, Borsello T, Gobbi M, Harris DA.An N-terminal fragment of the prion protein binds to amyloid-β oligomers and inhibits their neurotoxicity in vivo. J Biol Chem. 2013; 288:7857-66

Forloni G, Sclip A, Borsello T, Balducci C. The neurodegeneration in Alzheimer disease and the prion protein. Prion. 2013 Jan 1;7: 60-5 Esposito S, Pristerà A, Maresca G, Cavallaro S, Felsani A, Florenzano F, Manni L, Ciotti MT, Pollegioni L, Borsello T, Canu N.

Contribution of serine racemase/d-serine pathway to neuronal apoptosis. Aging Cell. 2012;11:588-98. Repici M, Chen X, Morel MP, Doulazmi M, Sclip A, Cannaya V, Veglianese P, Kraftsik R, Mariani J, Borsello T, Dusart I. Specific

inhibition of the JNK pathway promotes locomotor recovery and neuroprotection after mouse spinal cord injury. Neurobiol Dis. 2012;46:710-21.

Feligioni M, Brambilla E, Camassa A, Sclip A, Arnaboldi A, Morelli F, Antoniou X, Borsello T. Crosstalk between JNK and SUMO signalling pathways:deSUMOylation is protective agaist HO-induced cell injury. PLoS One. 2011;6(12):e28185. Epub 2011 Dec 2.

Sclip A, Antoniou X, Colombo A, Camici GG, Pozzi L, Cardinetti D, Feligioni M, Veglianese P, Bahlmann FH, Cervo L, Balducci C, Costa C, Tozzi A, Calabresi P, Forloni G, Borsello T. c-jun N-terminal kinase regulates soluble Abeta oligomers and cognitive impairment in AD mouse model . J Biol Chem. 2011 286:43871-80

Luigi Cervo, Ph.D. (Open University, Milton Keynes, U. K.), since 2006 is the head of the Experimental Psychopharmacology Laboratory. From 1978 to 2001 he was a research fellow and then chief of the Behavioural Pharmacology Unit in the Laboratory of Neuropharmacology and in 1981 he was awarded the degree in Biochemical Research from the “M. Negri” Institute. Between 1981 and 1983 he spent two years as a research fellow in the Department of Psychiatry at the Chicago University, Illinois, U.S.A (Prof. Charles Robert Schuster). His main research interests span the areas of behavioral neuroscience and psychopharmacology. They mainly focuses on experimental animal models and their translational application to complex human disorders such as drug abuse, anxiety and depression. Author and co-author of several peer-review articles, author of communications in international meetings, he is reviewer of several international peer-reviewed scientific journals. He is member of the Society for Neuroscience,

European Behavioural Pharmacological Society, Italian Society for Neuroscience and Italian Society of Neuropsychopharmacology. Selected publications

Grignaschi G, Burbassi S, Zennaro E, Bendotti C, Cervo L. A single high dose of cocaine induces behavioural sensitization and modifies mRNA encoding GluR1 and GAP-43 in rats. Eur J Neurosci 2004; 20: 2833-2837.

Cervo L, Canetta A, Calcagno E, Burbassi S, Sacchetti G, Caccia S, Fracasso C, Albani D, Forloni G, Invernizzi R. Deficits of serotonin synthesis cause resistance to antidepressants, J Neurosci 2005; 25: 8165-8172.

Cervo L, Cocco A, Petrella C, Heidbreder CA. Selective antagonism at dopamine D3 receptors attenuates cocaine seeking behaviour in the rat. Int J Neuropsychopharmacol. 2007; 10: 167-181.

Burbassi S, Cervo L. Stimulation of serotonin(2C) receptors influences cocaine-seeking behavior in response to drug-associated stimuli in rats. Psychopharmacology (Berl). 2008; 196: 15-27.

Burattini C, Burbassi S, Aicardi G, Cervo L. Effects of naltrexone on cocaine- and sucrose-seeking behaviour in response to associated stimuli in rats. Int J Neuropsychopharmacol. 2008; 11,: 103-109.

Fumagalli F, Franchi C, Caffino L, Racagni G, Riva MA, Cervo L. Single session of cocaine intravenous self-administration shapes goal-oriented behaviours and up-regulates Arc mRNA levels in rat medial prefrontal cortex. Int J Neuropsychopharmacol. 2009; 12: 423-429.

Watson J, Guzzetti S, Franchi C, Di Clemente A, Burbassi S, Emri Z, Leresche N, Parri HR, Crunelli V, Cervo L. Gamma-hydroxybutyrate does not maintain self-administration but induces conditioned place preference when injected in the ventral tegmental area. Int J Neuropsychopharmacol. 2010; 13:143-153.

Di Clemente A, Franchi C, Orrù A, Arnt J, Cervo L. Bifeprunox: a partial agonist at dopamine D2 and serotonin 1A receptors, influences nicotine-seeking behaviour in response to drug-associated stimuli in rats. Addict Biol. 2012; 17: 274-286.

Orrù A, Fujani D, Cassina C, Conti M, Di Clemente A, Cervo L. Operant, oral alcoholic beer self-administration by C57BL/6J mice: effect of BHF177, a positive allosteric modulator of GABA(B) receptors. Psychopharmacology (Berl). 2012; 222: 685-700.

Fumagalli F, Moro F, Caffino L, Orrù A, Cassina C, Giannotti G, Di Clemente A, Racagni G, Riva MA, Cervo L. Region-specific effects on BDNF expression after contingent or non-contingent cocaine i.v. self-administration in rats. Int J Neuropsychopharmacol. 2013; 16: 913-918.

Cervo L, Di Clemente A, Orrù A, Moro F, Cassina C, Pich EM, Corsi M, Gozzi A, Bifone A. Inhibition of glycine transporter-1 reduces cue-induced nicotine-seeking, but does not promote extinction of conditioned nicotine cue responding in the rat. Addict Biol. 2013; 18: 800-811.

Caffino L, Cassina C, Giannotti G, Orrù A, Moro F, Di Clemente A, Racagni G, Fumagalli F, Cervo L. Short-term abstinence from cocaine self-administration, but not passive cocaine infusion, elevates αCaMKII autophosphorylation in the rat nucleus accumbens and medial prefrontal cortex. Int J Neuropsychopharmacol. 2014; 17: 323-329.

Sukhanov I, Caffino L, Efimova EV, Espinoza S, Sotnikova TD, Cervo L, Fumagalli F, Gainetdinov RR. Increased context-dependent conditioning to amphetamine in mice lacking TAAR1. Pharmacol Res. 2015 ;103:206-214.

Roberto Chiesa graduated in Biological Sciences with major in Genetics at the University of Pavia in 1991, and obtained a Ph.D. in Pharmacology at the Mario Negri Institute for Pharmacological Research of Milan in 1994. From 1996 through 2000 he was Research Associate at the Department of Cell Biology and Physiology of Washington University in St. Louis, MO, USA. In 2001 Dr. Chiesa moved back to the Mario Negri Institute where he held a Telethon Scientist position (Dulbecco Telethon Institute, Telethon Foundation) until December 2013. Since 2009 is head of the Laboratory of Prion Neurobiology. His research activity is mainly focused on the molecular mechanisms of neuronal dysfunction and phenotypic heterogeneity in genetic prion diseases. Dr. Chiesa received the James L. O’Leary Prize (1998) and Bruno Ceccarelli Prize (2000) for research in neuroscience. He is member of editorial boards of PloS ONE and Biochemical Journal. Selected publications

Fioriti L, Dossena S, Stewart LR, Stewart RS, Harris DA, Forloni G, Chiesa R. Cytosolic prion protein (PrP) is not toxic in N2a cells and primary neurons expressing pathogenic PrP mutations. J Biol Chem. 280:11320-8 (2005)

Biasini E, Massignan T, Fioriti L, Rossi V, Dossena S, Salmona M, Forloni G, Bonetto V, Chiesa R Analysis of the cerebellar proteome in a transgenic mouse model of inherited prion disease reveals preclinical alteration of calcineurin activity. Proteomics. 6:2823-34 (2006)

Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron. 2008, 60:598-609 (2008).

Biasini E., Tapella L., Mantovani S., Stravalaci M., Gobbi M., Harris D.A. and Chiesa R. (2009) Immunopurification of pathological prion protein aggregates. PloS ONE, 4(11): e7816

Biasini E., Tapella L., Restelli E., Pozzoli M., Massignan T., and Chiesa R. (2010) The hydrophobic core region governs mutant prion protein aggregation and intracellular retention. Biochem Journal 430: 477-86

Restelli E., Fioriti L., Mantovani S., Airaghi S., Forloni G., and Chiesa R. (2010) Cell type-spcific neuroprotective activity of untranslocated prion protein. PloS ONE, 5(10): e13725

Quaglio E., Restelli E., Garofoli A., Dossena S., De Luigi A., Tagliavacca L., Imperiale D., Migheli A., Salmona M., Sitia R., Forloni G., and Chiesa R. (2011) Expression of mutant or cytosolic PrP in transgenic mice and cells is not associated with endoplasmic reticulum stress or proteasome dysfunction. PloS ONE, 6(4): e19339

Senatore A., Colleoni S., Verderio C., Restelli E., Morini R., Condliffe S.B., Bertani I., Mantovani S., Canovi M, Micotti E., Forloni G, Dolphin A.C., Matteoli M., Gobbi M., and Chiesa R. (2012) Mutant PrP suppresses glutamatergic neurotransmission in cerebellar granule neurons by impairing membrane delivery of VGCC 2-1 subunit. Neuron, 74: 300-313

Tapella L., Stravalaci M., Bastone A., Biasini E., Gobbi M. and Chiesa R. (2013) Epitope scanning indicates structural differences

in brain-derived monomeric and aggregated mutant prion proteins related to genetic prion diseases. Biochem J. 454: 417-425

Alvarez-Laviada A., Kadurin I., Senatore A., Chiesa R., and Dolphin A.C. (2014). The inhibition of functional expression of calcium channels by prion protein demonstrates competition with 2 for GPI-anchoring pathways. Biochem. J. 458: 365-374

Bouybayoune I., Mantovani S., Del Gallo F., Bertani I., Restelli E., Tapella L., Comerio L., Bianchi S., Fernández-Borges N., Mangieri M., Bisighini C., Beznoussenko G.V., Paladini A., Balducci C., Micotti E., Forloni G., Castilla J., Fiordaliso F., Tagliavini F., Imeri L., Chiesa R. (2015). Transgenic fatal familial insomnia mice indicate prion infectivity-independent mechanisms of pathogenesis and phenotypic expression of disease. PLoS Pathogens, 11(4):e1004796

Chiesa R. (2015). The elusive role of the prion protein and the mechanism of toxicity in prion disease. PLoS Pathogens 11(5):e1004745

Barbara D’Avanzo obtained her master in philosophy at the University of Milan in 1989. Her main field of interest is epidemiologic research in mental health and quality evaluation of the mental health services. First involved in the analysis of the implementation of the psychiatric reform in Italy, then addressed the quality and the role of residential facilities and treatment and continuity of care in the community services network. She works at the effectiveness evaluation and implementation problems of the most common psychosocial and psychological interventions for severe mental illness. She e has implemented a monitoring system of suicide attempts and self-harm episodes in various areas of Italy, in the framework of suicide mortality monitoring and suicide prevention study and implementation, and is also working on issues related to recovery-oriented services, consumers’ empowerment, methods of consumers participation to service evaluation, and acknowledgment of the value of consumers’ knowledge and perspective about mental health services and treatments. Activity of education and training in the mental health services and mental health literacy to the organizations active in the community. She is head of the Laboratory of Epidemiology and Social Psychiatry since 2011, and is member of the Scientific National Board of the World Association for Psychosocial Rehabilitation. Selected publications

Barbato A, D'Avanzo B, D'Anza V, Montorfano E, Savio M, Corbascio G C. Involvement of users and relatives in mental health service evaluation. J Nerv Ment Dis 2014 202 : 479-486.

Barbato A, Bajoni A, Rapisarda F, D'Anza V, De Luca L F, Inglese C, Japichino S, Mauriello F, D'Avanzo B. Quality assessment of mental health care by people with severe mental disorders: a participatory research project. Community Ment Health J 2014 50 : 402-408

Barbato A, Bossini L, Calugi S, D'Avanzo B, Fagiolini A, Koukouna D, Parabiaghi A, Rapisarda F, Rucci P, Vallarino M.Validation of the Italian version of the Functioning Assessment Short Test (FAST) for bipolar disorder. Epidemiol Psychiatr Sci. 2013; 22:187-94

D'Avanzo B, Barbato A, Erzegovesi S, Lampertico L, Rapisarda F, Valsecchi L Formal and informal help-seeking for mental health problems. A survey of preferences of italian students. Clin Pract Epidemiol Ment Health. 2012;8:47-51

Parabiaghi A, D'Avanzo B, Tettamanti M, Barbato A, GISAS Study Group. The GiSAS study. Rationale and design of a pragmatic randomized controlled trial on aripiprazole, olanzapine and haloperidol in the long-term treatment of schizophrenia. Contemp Clin Trials 2011; 32:675-684.

Barbato A, Parabiaghi A, Panicali F, Battino N, D'Avanzo B, De Girolamo G, Rucci P, Santone G, PROGRES-Acute Group. Do patients improve after short psychiatric admission? A cohort study in Italy Nord J Psychiatry 2010; E-pub

Campi R, Barbato A, D'Avanzo B, Guaiana G, Bonati M Suicide in Italian children and adolescents J Affect Disord 2009; 113:291-295. Barbato A, D'Avanzo B. Efficacy of couple therapy as a treatment for depression: a meta-analysis. Psychiatr Q 2008; 79:121-132. D'Avanzo B, Aliprandini E, Beghi M, Cornaggia C M, Erlicher A, Frova M, Mascarini A, Miragoli P, Righi A. Strutture residenziali e

semiresidenziali nei servizi di salute mentale. Dove sta la differenza? Epidemiologia e Psichiatria Sociale 2008; 17:57-64. Barbato A, D'Avanzo B. Marital therapy for depression. Cochrane Database Systematic Reviews 2006; Issue 2. Parabiaghi A, Barbato A, D'Avanzo B, Erlicher A, Lora A. Assessing reliable and clinically significant change on Health of the Nation

Outcome Scales: method for displaying longitudinal data. Aust N Z J Psychiatry 2005; 39:719-725. Barbato A, D'Avanzo B. Involuntary placement in Italy. Br J Psychiatry 2005; 186:542-543. Guaiana G, Andretta M, Corbari L, Mirandola M, Sorio A, D'Avanzo B, Barbui C. Antidepressant drug consumption and public health

indicators in Italy, 1955-2000. J Clinical Psychiatry 2005; 66:750-755. D'Avanzo B, Battino R N, Gallus S, Barbato A. Factors predicting discharge of patients from community residential facilities: A

longitudinal study from Italy. Aust N Z J Psychiatry 2004; 38:619-628. D'Avanzo B, Barbato A, Barbui C, Battino N, Civenti G, Frattura L. Discharges of patients from public psychiatric hospitals in Italy

between 1994 and 2000. Int J Social Psychiatry 2003; 49 27-3.

Maria Grazia De Simoni received an Honours Doctoral Degree in Biological Sciences from the University of Milan, Italy I 1977 and subsequently a PhD degree in Neuropharmacology from the Mario Negri Institute of Milan. Having been awarded a European Community fellowship for "Advanced Professional Training", she worked as a post-doc at INSERM U 171, Université Claude Bernard, Lyon, France on the neurochemistry of sleep. Presently she is the Head of the Laboratory of Inflammation and Nervous System Diseases, Mario Negri Institute Milan, Italy. Scientific activity. Long standing experience on experimental models of brain diseases, including Alzheimer’s disease, epilepsy, stroke and trauma. Presently her main scientific interests include the study of the pathogenesis of cerebral ischemia and traumatic brain injury and the identification of molecular mechanisms and novel protective strategies, with particular focus on inflammation and immune system contribution to CNS conditions. She has demonstrated the pathogenetic role of the complement system in acute brain injury and the neuroprotective effects of complement inhibitors. She held more than 100 lectures in Italy, United States and Australia. She is the Author of more than 140 scientific papers on peer-reviewed international journals. Official H index: 40. Editorial activity. Board memberships: Senior Editor of

Stroke, American Heart Association, 2007-2010; Intensive Care Medicine experimental, Senior Editor, SRN Vascular Medicine; The Open Pathology Journal; Frontiers in Immunology: Frontiers in Molecular Innate Immunity. Selected publications

Zanier E R, Marchesi F, Ortolano F, Perego C, Arabian M, Zoerle T, Sammali E, Pischiutta F, De Simoni MG. Fractalkine receptor

deficiency is associated with early protection, but late worsening of outcome following brain trauma in mice. J Neurotrauma 2015; Epub. Fumagalli S, Perego C, Pischiutta F, Zanier ER, De Simoni MG. The ischemic environment drives microglia and macrophage function.

Front Neurol 2015; 6: 81. Llovera G, Hofmann K, Roth S, Salas-Perdomo A, Ferrer-Ferrer M, Perego C, Zanier E R, Mamrak U, Rex A, Party H, Agin V, Fauchon

C, Orset C, Haelewyn B, De Simoni MG, Dirnagl U, Planas A M, Plesnila N, Vivien D, Liesz A. Results of a multicenter trial in experimental stroke research: Anti-CD49d treatment for acute brain ischemia. Sci Transl Med 2015; 7(299): 299ra121.

Fumagalli S, Ortolano F and De Simoni MG. A close look at brain dynamics: cells and vessels seen by in vivo two-photon microscopy. Prog. Neurobiol. 2014; 21: 36-54.

Longhi L, Orsini F, De Blasio D, Fumagalli S, Ortolano F, Locatelli M, Stocchetti N, De Simoni M G. Mannose binding lectin expressed after clinical and experimental traumatic brain injury and its deletion is protective. Critical Care Medicine, 2014; 24: 1910-1918.

Pischiutta F, D’Amico G, Dander E, Biondi A, Biagi E, Citerio G, De Simoni MG, Zanier ER. Immunosuppression does not affect human bone marrow mesenchymal stromal cell efficacy after transplantation in traumatized mice brain. Neuropharmacology 2014, 79: 119-126.

Zanier ER, Zangari R, Munthe-Fog L, Hein E, Zoerle T, Conte V, Orsini F, Tettamanti M, Stocchetti N, Garred P, De Simoni MG. Ficolin-3 mediated lectin complement pathway activation in subarachnoid hemorrhage patients. Neurology 2014; 82,1-9..

Longhi L, Perego C, Ortolano F, Aresi S, Fumagalli S, Zanier E R, Stocchetti N, De Simoni MG. Tumor necrosis factor in traumatic brain injury: effects of genetic deletion of p55 or p75 receptor. J Cereb Blood Flow Metab 2013, 33: 1182-1189.

Fumagalli S, Perego C, Ortolano F, De Simoni MG. CX3CR1 deficiency induces an early protective M2 microglia/macrophage polarization in ischemic mice. Glia 2013; 61: 827-842.

Orsini F, Villa P, Parrella S, Zangari R, Zanier E, Gesuete R, Stravalaci M, Ottria R, Reina JJ, Paladini A, Micotti E, Ribeiro-Viana R, Rojo J, Pavlov VI, Stahl GL, Bernardi A, Gobbi M, and De Simoni MG. Targeting mannose binding lectin confers long lasting protection with a surprisingly wide therapeutic window in cerebral ischemia. Circulation 2012; 126: 1484-1494.

Helmy A, De Simoni MG, Guilfoyle MR, Carpenter KL, Hutchinson PJ. Cytokines and innate inflammation in the pathogenesis of human traumatic brain injury. Prog Neurobiol 2011; 95: 352-372.

Gesuete R, Storini C, Fantin A, Stravalaci M, Zanier ER, Orsini F, Vietsch H, Mannesse MLM, Ziere B, Gobbi M, De Simoni MG. Recombinant C1-inhibitor in Brain Ischemic Injury. Annals Neurol 2009; 66: 332-342

Roberto W. Invernizzi got his Biological Sciences degree at the Università Statale di Milano in 1986. He spent short periods at the Department of Pharmacology of the Karolinska Institutet, Stockholm (1988) and Nihon University, Tokyo (1995) where he consolidated his knowledge on the intracerebral microdialysis technique in the rat. In 1996 he was appointed Head of the Intracerebral Microdialysis Unit and from 2006 Head of the Laboratory of Neurochemistry and Behavior of the I.R.F. “Mario Negri”. Scientific Interests: Biological bases of psychotropic drugs action and role of brain neurotransmitters and circuits in cognitive symptoms of neuropsychiatric diseases. Recently, the laboratory focused on the development of experimental models of Rett syndrome (RTT) in mice with the aim of studying the underlying pathogenic mechanisms and identifying biological targets to develop novel therapeutics. He is member of the Società Italiana di Neuroscienze, and Società Italiana di Farmacologia, AIRETT Research Team, editorial board of the J. Neurochemistry and reviewer for several International journals in the field of pharmacology and neurochemistry. He published more than 80 scientific articles in peer-reviewed journals and some book chapters.

Selected publications Carli M, Kostoula C, Sacchetti G, Mainolfi P, Anastasia A, Villani C, Invernizzi RW. Tph2 gene deletion enhances amphetamine-induced

hypermotility: effect of 5-HT restoration and role of striatal noradrenaline release. J Neurochem 2015, 135:674-685 Macrì S, Ceci C, Proietti Onori M, Invernizzi RW, Bartolini E, Altabella L, Canese R, Imperi M, Orefici G, Creti R, Margarit I, Magliozzi

R Laviola G. Mice repeatedly exposed to group-A b-Haemolytic Streptococcus show perseverative behaviors, impaired sensori motor gating, and immune activation in rostral diencephalon. Scientific Rep. 2015, 5:13257

Batelli S, Invernizzi RW, Negro A, Calcagno E, Rodilossi S, Forloni G, Albani D. The Parkinson's Disease-Related Protein DJ-1 Protects Dopaminergic Neurons in vivo and Cultured Cells from Alpha-Synuclein and 6-Hydroxydopamine Toxicity. Neurodegener Dis. 2015, 15:13-23

Carli M and Invernizzi RW. Serotoninergic and dopaminergic modulation of cortico-striatal circuit in executive and attention deficits induced by NMDA receptor hypofunction in the 5-choice serial reaction time task. Frontiers in Neural Circuits 8: 58. doi: 10.3389/fncir.2014.00058

Agnoli L, Mainolfi P, Invernizzi RW and Carli M. Dopamine D1-Like and D2-Like Receptors in the Dorsal Striatum Control Different Aspects of Attentional Performance in the Five-Choice Serial Reaction TimeTask Under a Condition of Increased Activity of Corticostriatal Inputs. Neuropsychopharmacology 2013, 38:701-714

Revel FG, Meyer CA, Bradaia A, Jeanneau K, Calcagno E, Andre´ CB, Haenggi M, Miss M-T, Galley G, Norcross RD, Invernizzi RW, Wettstein JG, Moreau J-L and Hoener MC. Brain-Specific Overexpression of Trace Amine-Associated Receptor 1 Alters Monoaminergic Neurotransmission and Decreases Sensitivity to Amphetamine. Neuropsychopharmacology 2012, 37: 2580-92

Cristini S, Alessandri G, Acerbi F, Ciusani E, Colombo A, Fascio U, Nicosia R, Invernizzi RW, Gelati M, Parati EA, Invernici G. 3-DSelf-organizing Neural Architectures: a Neural Stem Cells reservoir and a system for neurodevelopmental studies. Tissue Engineering,Part C 2011, 17:1109-1120.

Carli M, Calcagno E, Mainolfi P, Mainini E, Invernizzi RWEffects of aripiprazole, olanzapine, and haloperidol in a model of cognitivedeficit of schizophrenia in rats: relationship with glutamate release in the medial prefrontal cortex. Psychopharmacology (Berl) 2011, 214 :639-652

Pozzi L, Baviera M, Sacchetti G, Calcagno E, Balducci C, Invernizzi RW, Carli M Attention deficit induced by blockade of N-methyl D-

Aspartate receptors in the prefrontal cortex is associated with enhanced glutamate release and cAMP response element binding proteinphosphorylation: role of metabotropic glutamate receptors 2/3. Neuroscience 2011, 176 : 336-348

D'Alessandro G, Calcagno E, Tartari S, Rizzardini M, Invernizzi RW, Cantoni L Glutamate and glutathione interplay in a motor neuronalmodel of amyotrophic lateral sclerosis reveals altered energy metabolism. Neurobiol Dis 2011, 43 : 346-355

Ugo Lucca got his Master of Science, University of Aberdeen - UK, 1999. At the Mario Negri Institute he was investigator from 1986- 1995, head of the "Clinical Evaluation of Antidementia Drugs Unit" (1995-1996) and, since 1996, head of the "Laboratory of Geriatric Neuropsychiatry". The main areas of interests include epidemiology and clinic features of dementia; natural history of dementia; neuropsychiatric disorders of the elderly; instruments for the screening diagnosis and clinical course assessment of dementia; clinical evaluation of anti dementia treatments and CNS active drugs (phase I, II, III, IV and observational studies). Selected publications

Lucca U, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Riva E. Association of Mild Anemia with Cognitive, Functional, Mood and Quality of Life Outcomes in the Elderly: The “Health and Anemia” Study. PlOS ONE 2008;3(4):e1920

Riva E, Tettamanti M, Mosconi P, Apolone G, Gandini F, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Guala A, Fasolo G, Lucca U. Association of mild anemia with hospitalization and mortality in the Elderly: The Health and Anaemia Population-based Study. Haematologica 2009;94:22-28

• Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, Nobili A, Tallone MV, Detoma P, Giacomin A, Clerico M, Tempia P, Savoia L, Fasolo G, Ponchio L, Della Porta MG, Riva E. Prevalence, incidence and types of mild anemia in the elderly: the “Health and Anemia”population-based study. Haematologica 2010; 95: 1849-1856.

Franchi C, Lucca U, Tettamanti M, Riva E, Fortino I, Bortolotti A, Merlino L, Pasina L, Nobili A. Cholinesterase inhibitor use in Alzheimer's disease: the EPIFARM-Elderly Project. Pharmacoepidemiol Drug Saf 2011; 20: 497-505.

Lucca U, Garrì M, Recchia A, Logroscino G, Tiraboschi P, Franceschi M, Bertinotti C, Biotti A, Gargantini E, Maragna M, Nobili A, Pasina L, Franchi C, Riva E, Tettamanti M. A Population-based study of dementia in the oldest old: the Monzino 80-plus study. BMC Neurology 2011; 11: 54.

Albani D, Tettamanti M, Batelli S, Polito L, Dusi S, Ateri E, Forloni G, Lucca U. Interleukin-1a, Interleukin-1b and Tumor Necrosis Factor-a genetic variants and risk of dementia in the very old: evidence from the “Monzino 80-plus” prospective study. Age (Dordr). 2012; 34:519-26

• Merlo A, Zemp D, Zanda E, Rocchi S, Meroni F, Tettamanti M, Recchia A Lucca U, Quadri P. Postural stability and history of falls in cognitively able older adults: the Canton Ticino study. Gait Posture. 2012; 36:662-6

Pasina L, Djade CD, Lucca U, Nobili A, Tettamanti M, Franchi C, Salerno F, Corrao S, Marengoni A, Iorio A, Marcucci M, Violi F, Mannucci PM. Association of anticholinergic burden with cognitive and functional status in a cohort of hospitalized elderly: comparison of the anticholinergic cognitive burden scale and anticholinergic risk scale: results from the REPOSI study. Drugs Aging. 2013; 30:103-12.

Polito L, Kehoe PG, Davin A, Benussi L, Ghidoni R, Binetti G, Quadri P, Lucca U, Tettamanti M, Clerici F, Bagnoli S, Galimberti D, Nacmias B, Sorbi S, Guaita A, Scarpini E, Mariani C, Forloni G, Albani D. The SIRT2 polymorphism rs10410544 and risk of Alzheimer's disease in two Caucasian case-control cohorts. Alzheimers Dement 2013;9(4):392-399.

Haïk S, Marcon G, Mallet A, Tettamanti M, Welaratne A, Giaccone G, Azimi S, Pietrini V, Fabreguettes JR, Imperiale D, Cesaro P, Buffa C, Aucan C, Lucca U, Peckeu L, Suardi S, Tranchant C, Zerr I, Houillier C, Redaelli V, Vespignani H, Campanella A, Sellal F, Krasnianski A, Seilhean D, Heinemann U, Sedel F, Canovi M, Gobbi M, Di Fede G, Laplanche JL, Pocchiari M, Salmona M, Forloni G, Brandel JP, Tagliavini F. Doxycycline in Creutzfeldt-Jakob disease: a phase 2, randomised, double-blind, placebo-controlled trial. Lancet Neurol 2014;13(2):150-158.

Lawlor B, Kennelly S, O'Dwyer S, Cregg F, Walsh C, Coen R, Kenny RA, Howard R, Murphy C, Adams J, Daly L, Segurado R, Gaynor S, Crawford F, Mullan M, Lucca U, Banzi R, Pasquier F, Breuilh L, Riepe M, Kalman J, Wallin A, Borjesson A, Molloy W, Tsolaki M, Olde Rikkert M. NILVAD protocol: a European multicentre double-blind placebo-controlled trial of nilvadipine in mild-to-moderate Alzheimer's disease. BMJ Open 2014 9;4(10):e006364. doi: 10.1136/bmjopen-2014-006364.

Piscopo P, Tosto G, Belli C, Talarico G, Galimberti D, Gasparini M, Canevelli M, Poleggi A, Crestini A, Albani D, Forloni G, Lucca U, Quadri P, Tettamanti M, Fenoglio C, Scarpini E, Bruno G, Vanacore N, Confaloni A. SORL1 Gene is Associated with the Conversion from Mild Cognitive Impairment to Alzheimer's Disease. J Alzheimers Dis 2015;46(3):771-776.

Forloni G, Tettamanti M, Lucca U, Albanese Y, Quaglio E, Chiesa R, Erbetta A, Villani F, Redaelli V, Tagliavini F, Artuso V, Roiter I. Preventive study in subjects at risk of fatal familial insomnia: Innovative approach to rare diseases. Prion 2015;9(2):75-79.

Lucca U, Tettamanti M, Logroscino G, Tiraboschi P, Landi C, Sacco L, Garrì M, Ammesso S, Bertinotti C, Biotti A, Gargantini E, Piedicorcia A, Nobili A, Pasina L, Franchi C, Djade CD, Riva E, Recchia A. Prevalence of dementia in the oldest old: the Monzino 80-plus population based study. Alzheimers Dement 2015;11(3):258-270.

Mandelli S, Riva E, Tettamanti M, Detoma P, Giacomin A, Lucca U. Mortality Prediction in the Oldest Old with Five Different Equations to Estimate Glomerular Filtration Rate: The Health and Anemia Population-based Study. PLoS One 2015;10(8):e0136039. doi: 10.1371/journal.pone.0136039.

Alessandro Nobili got his degree in Medicine (Milan, 1990). Master in Biotechonological Research, Regione Lombardia, Milan 1988. International School of Pharmacology, 31° Course on: Drug Epidemiology and Post-marketing Surveillance, Erice, September 1990. Course on: Methods in Epidemiological Research, Milan, October 1990. Course: Long Term Clinical Trials, Cogne January 1991. Main areas of interest Methodology of Randomized Clinical Trials; Pharmacoepidemiology and post-marketing surveillance research; Drug utilization studies; Quality assessment of geriatric services; Qualitative studies on caregiver role in the care of patients with dementia; Methodological evaluation of the Special Care Unit for Alzheimer Disease patients; Methodology of drug information. Employment and research experience Chief of the Unit of Quality Assessment of Geriatric Services Chief of the Drug Information Services for the Elderly, Laboratory of Geriatric Neuropsychiatry, Istituto di Ricerche Farmacologiche “Mario Negri”, Milan. Editorial Board of the MICROMEDEX

Inc., Englewood, Colorado 80111-4740 USA. National Expert accredited by Italian Ministry of Health for The Italian (AIFA) and European Agency for the Evaluation of Medicinal Products (EMEA). Head of the Laboratory of the Quality Assessment of Geriatric Services at the Mario Negri Institute since 2007. Selected publications

Franchi C, Ardoino I, Rossio R, Nobili A, Biganzoli EM, Marengoni A, Marcucci M, Pasina L, Tettamanti M, Corrao S, Mannucci PM;

REPOSI Investigators. Prevalence and Risk Factors Associated with Use of QT-Prolonging Drugs in Hospitalized Older People. Drugs Aging. 2016 Jan;33(1):53-61. doi: 10.1007/s40266-015-0337-y. PubMed PMID: 26693921.

Marengoni A, Nobili A, Onder G. Best Practices for Drug Prescribing in Older Adults: A Call for Action. Drugs Aging. 2015 Nov;32(11):887-90. doi: 10.1007/s40266-015-0324-3.

Riva S, Nobili A, Djade CD, Mancuso ME, Santagostino E, Pravettoni G. Cognitive and psychological profiles in treatment compliance: a study in an elderly population with hemophilia. Clin Interv Aging. 2015 Jul 9;10:1141-6. doi: 10.2147/CIA.S84749. eCollection 2015. PubMed PMID: 26185433; PubMed Central PMCID: PMC4501679.

Bellelli G, Nobili A, Annoni G, Morandi A, Djade CD, Meagher DJ, Maclullich AM, Davis D, Mazzone A, Tettamanti M, Mannucci PM; REPOSI (REgistro POliterapie SIMI) Investigators. Under-detection of delirium and impact of neurocognitive deficits on in-hospital mortality among acute geriatric and medical wards. Eur J Intern Med. 2015 Nov;26(9):696-704. doi: 10.1016/j.ejim.2015.08.006. Epub 2015 Aug 31. PubMed PMID: 26333532.

Bazzoni G, Marengoni A, Tettamanti M, Franchi C, Pasina L, Djade CD, Fortino I, Bortolotti A, Merlino L, Nobili A. The drug prescription network: a system-level view of drug co-prescription in community-dwelling elderly people. Rejuvenation Res. 2015 Apr;18(2):153-61. doi: 10.1089/rej.2014.1628. PubMed PMID: 25531938.

Mannucci PM, Nobili A; REPOSI Investigators. Multimorbidity and polypharmacy in the elderly: lessons from REPOSI. Intern Emerg Med. 2014 Oct;9(7):723-34. doi: 10.1007/s11739-014-1124-1. Epub 2014 Aug 28. PubMed PMID: 25164413.

Pasina L, Djade CD, Nobili A, Tettamanti M, Franchi C, Salerno F, Corrao S, Marengoni A, Iorio A, Marcucci M, Mannucci P. Drug-drug interactions in a color of hospitalized elderly patients. Pharmacoepidemiol Drug Saf. 2013 Oct;22(10):1054-60. doi: 10.1002/pds.3510. Epub 2013 Aug 30. PubMed PMID: 24038765.

Nobili A, Marengoni A, Tettamanti M, Salerno F, Pasina L, Franchi C, Iorio A, Marcucci M, Corrao S, Licata G, Mannucci P M. Association between clusters of diseases and polypharmacy in hospitalized elderly patients: Results from the REPOSI study. Eur J Intern Med 2011;22:597-602.

Nobili A, Franchi C, Pasina L, Tettamanti M, Baviera M, Monesi L, Roncaglioni C, Riva E, Lucca U, Bortolotti A, Fortino I, Merlino L. Drug utilization and polypharmacy in an Italian elderly population: the EPIFARM-elderly project. Pharmacoepidemiol Drug Saf. 2011 May;20(5):488-96. doi: 10.1002/pds.2108. Epub 2011 Jan 24. PubMed PMID: 21264988.

Nobili A, Licata G, Salerno F, Pasina L, Tettamanti M, Franchi C, De Vittorio L, Marengoni A, Corrao S, Iorio A, Marcucci M, Mannucci PM; SIMI Investigators. Polypharmacy, length of hospital stay, and in-hospital mortality among elderly patients in internal medicine wards. The REPOSI study. Eur J Clin Pharmacol. 2011 67:507-19.

Annamaria Vezzani got her Degree in Biological Science at the University of Milan in 1978 and she specialized in Neuropharmacology at the Mario Negri Institute in 1982. She spent her post-doctoral period in Baltimore at the University of Maryland in 1983-1984 working on the mechanisms of epileptogenesis in experimental models of epilepsy. She spent additional post-doctoral periods at the University of Stockholm and at the Karolinska Institute between 1985 and 1999. She was on sabbatical at the Albert Einstein College of Medicine in 2002 in the laboratory of Developmental Epilepsy. She is involved in studies on the biochemical and molecular mechanisms involved in the etiopathogenesis of seizures disorders using experimental models of epilepsy. The present research is focused on the functional role of neuroactive peptides and inflammatory mediators in the modulation of neuronal excitability and seizure-related neurodegeneration. Focus of the research is also on the mechanisms of pharmacoresistance. Since 1997 she is the Head of the Laboratory of Experimental Neurology at the Mario Negri Institute. She is member of the Editorial Board of various scientific journals and Associate Editor for basic science of Epilepsia, the official journal of the International League Against Epilepsy (ILAE). She has been appointed of the Chair of the Commission on Neurobiology of ILAE which is promoting initiatives for improving translational research in epilepsy. She has been awarded of the prestigeous Epilepsy Research Recognition Award for translational research in 2009 by the American Epilepsy Society

Selected publications

Vezzani A, Conti M, De Luigi A, Ravizza T, Moneta D, Marchesi F, De Simoni MG. Interleukin-1beta immunoreactivity and microglia

are enhanced in the rat hippocampus by focal kainate application: functional evidence for enhancement of electrographic seizures.(1999) J Neurosci.19:5054-65.

Vezzani A., Moneta D., Conti M., Richichi C., Ravizza T., De Luigi A., De Simoni M.G., Sperk, Andell-Jonsson S., Lundkvist J., Iverfeldt K. and Bartfai T. Powerful anticonvulsant action of IL-1 receptor antagonist upon intracerebral injection and astrocytic overexpression in mice (2000) Proc Natl Acad Sci USA, 97: 11534

Rizzi M, Caccia S, Guiso G, Richichi C, Gorter JA, Aronica E, Aliprandi M, Bagnati R, Fanelli R, D'Incalci M, Samanin R, Vezzani A. Limbic seizures induce P-glycoprotein in rodent brain: functional implications for pharmacoresistance (2002) J Neurosci, 22: 5833

Balosso S, Ravizza T, Perego C, Peschon J, Campbell I, De Simoni MG, Vezzani A. TNF-alpha inhibits kainic acid-induced seizures in mice via p75 receptors (2005) Ann Neurol, 57: 804-12

Ravizza T, Gagliardi B, Noè F, Boer K, Aronica E and Vezzani A. Innate and adaptive immunity during epiletogenesis and spontaneous seizures: evidence from experimental models and human temporal lobe epilepsy (2008) Neurobiol Dis, 29: 142

Noè F, Pool AH, Nissinen J, Gobbi M, Bland R, Rizzi M, Balducci C, Ferraguti F, Sperk G, During MJ, Pitkänen A, Vezzani A. Neuropeptide Y gene therapy decreases chronic spontaneous seizures in a rat model of temporal lobe epilepsy (2008) Brain, 131:1506

Balosso S, Maroso M, Sanchez-Alavez M, Ravizza T, Frasca A, Bartfai T, Vezzani A. A novel non-transcriptional pathway mediates the proconvulsive effects of interleukin-1beta. (2008) Brain, 131:3256

Maroso M, Balosso S, Ravizza T, Liu J, Aronica E, Iyer A, Rossetti C, Molteni M, Casalgrandi M, Manfredi AA, Bianchi ME and Vezzani A. Toll-Like Receptor 4 (TLR4) and High Mobility Group Box 1 (HMGB1)are involved in ictogenesis and can be targeted to reduce seizures (2010) Nature Medicine, 16:413-9.

Vezzani A, French J, Bartfai T, Baram TZ. The role of inflammation in epilepsy. (2011) Nat Rev Neurol, 7: 31-40. Nabbout R, Vezzani A, Dulac O, Chiron C. Acute encephalopathy with inflammation-mediated status epilepticus. (2011) Lancet Neurol.

10:99-108 Vezzani A. Before epilepsy unfolds: finding the epileptogenesis switch. Nat Med. 2012; 18:1626-7 Vezzani A Fetal brain inflammation may prime hyperexcitability and behavioral dysfunction later in life. Ann Neurol. 2013; 74:1-3 Balosso S, Liu J, Bianchi ME, Vezzani A. (2014) Disulfide-containing HMGB1 promotes NMDA receptor function and excitotoxicity by

activating TLR4-dependent signaling in hippocampal neurons. Antioxid Redox Signal, 21: 1726-40 PMCID24094148 Baron R, Ferriero DM, Frisoni GB, Bettegowda C, Gokaslan ZL, Kessler JA, Vezzani A, Waxman SG, Jarius S, Wildemann B, Weller

M.Neurology-the next 10 years. Nat Rev Neurol. 2015; 11:658-64

Diego Albani graduated in Biology in 1996 (110/110 cum laude), and attended the course of Specialist in Pharmacological Research at the Mario Negri where he operates since 2002, after a 3-year period of post-doctoral carried out in the laboratory of Prof. Renato Dulbecco at CNR-ITBA Milan. He became head of the Unit of Genetics of Neurodegenerative Disorders in 2011. His current interests include the biological basis of neurodegenerative diseases, including Alzheimer's disease (AD), Parkinson's disease (PD) and frontotemporal dementia (FTLD) , with particular attention to genetic aspects and oxidative stress. Dr. Albani is actively involved in research projects concerning topics of pharmacogenomics of AD, the development of biomaterials for drug delivery and modulation of neuronal enzymes of the family of sirtuins (SIRT1 and SIRT2) as a possible new strategy against AD and PD. He is currently a member of the Editorial Board of the Journal of Alzheimer's Disease and Associated Editor of Frontiers. Selected pubblications

Albani D, Zucchi I, Bini L, Valaperta R, Liberatori S, Montagna C, Susani L, Barbieri O, Pallini V, Vezzoni P, Dulbecco R “Dome formation in cell cultures as expression of an early stage of lactogenic differentiation of the mammary gland “ Proc Natl Acad Sci USA, 2002; 99:8660-5.

Albani D, Peverelli E, Rametta R, Batelli S, Veschini L, Negro A, Forloni G : “Protective effect of TAT-delivered a-synuclein: relevance of the C-terminal domain and involvement of HSP70” FASEB J, 2004; 18:1713-1715.

Albani D, Roiter I, Artuso V, Batelli S, Prato F, Pesaresi M, Galimberti D, Scarpini E, Bruni A, Franceschi M, Piras MR, Confaloni A, Forloni G “Presenilin-1 mutation E318G and familial Alzheimer's disease in the italian population”. Neurobiol Aging, 2007; 28:1682-8.

Albani D., Polito L., Batelli S., De Mauro S., Fracasso C., Martelli G., Colombo L., Manzoni C., Salmona M., Caccia S., Negro A., Forloni G The SIRT1 activator resveratrol protects SK-N-BE cells from oxidative stress and against toxicity caused by α-synuclein or amyloid-β (1-42) peptide. J Neurochem, 2009; 110:1445-56.

Batelli S, Peverelli E, Rodilossi S, Forloni G, Albani D. Macroautophagy and the proteasome are differently involved in the degradation of alpha-synuclein wild type and mutated A30P in an in vitro inducible model (PC12/TetOn). Neuroscience, 2011; 195:128-37.

Albani D, Tettamanti M, Batelli S, Polito L, Dusi S, Ateri E, Forloni G, Lucca U (2011) Interleukin-1a, Interleukin-1b and Tumor Necrosis Factor-a genetic variants and risk of dementia in the very old: evidence from the “Monzino 80-plus” prospective study. Age (Dordr). 2012; 34:519-26.

Martinelli-Boneschi F, Giacalone G, Magnani G, Biella G, Coppi E, Santangelo R, Brambilla P, Esposito F, Lupoli S, Clerici F, Benussi L, Ghidoni R, Galimberti D, Squitti R, Confaloni A, Bruno G, Pichler S, Mayhaus M, Riemenschneider M, Mariani C, Comi G, Scarpini E, Binetti G, Forloni G, Franceschi M, Albani D. Pharmacogenomics in Alzheimer's disease: a genome-wide association study of response to cholinesterase inhibitors. Neurobiol Aging. 2013; 34:1711.e7-13.

Polito L, Kehoe PG, Davin A, Benussi L, Ghidoni R, Binetti G, Quadri P, Lucca U, Tettamanti M, Clerici F, Bagnoli S, Galimberti D, Nacmias B, Sorbi S, Guaita A, Scarpini E, Mariani C, Forloni G, Albani D. The SIRT2 polymorphism rs10410544 and risk of Alzheimer's disease in two Caucasian case-control cohorts. Alzheimers Dement. 2013; 9:392-9.

Tunesi M, Prina E, Munarin F, Rodilossi S, Albani D, Petrini P, Giordano C. Cross-linked poly(acrylic acids) microgels and agarose as semi-interpenetrating networks for resveratrol release. J Mat Sci Mat Med 2015;26:5328.

Batelli S, Invernizzi RW, Negro A, Calcagno E, Rodilossi S, Forloni G, Albani D. The Parkinson’s disease-related protein DJ-1 protects dopaminergic neurons in vivo and cultured cells from alpha-synuclein and 6-hydroxydopamine toxicity. Neurodegenerative Diseases, 2015;15:13-23.

Mirjana Carli started her scientific career in the laboratory of Neuropharmacology of the “Istituto di Ricerche Farmacologiche Mario Negri” Milan in 1977, where, at present, she heads the Pharmacology of Cognitive Behaviour Unit. From 1981 to 1985 she worked in the laboratory of Cognitive Neuroscience, Dept. of Experimental Psychology, University of Cambridge (UK) directed by Prof. Trevor W. Robbins. Here she took interest in the role of brain monoamines in attention and response control, and for this purpose developed several behavioural tests for rats. In 1986 she returned to the laboratory of Neuropharmacology of the “Istituto di Ricerche Farmacologiche Mario Negri” and from 2006 in the laboratory of Neurochemistry and Behaviour were she leads the Unit of Pharmacology of Cognitive Behaviour. Here she devoted her efforts to the study of serotonin receptors in emotional behaviour and cognitive processes such as learning, memory and attention. She has made significant contribution to understanding the role of some serotonin receptors in the control of executive processes and attention. Recently the unit has expanded its interest to genetic bases of behaviour and cognition with a focus on Rett syndrome. In 2005 she obtained the PhD from the Open University (UK). She is member of The Society of Neuroscience and of European Behavioural Pharmacology Society (EBPS) and peer reviewer for several international journals in the field of neuropsychopharmacology and behavioural neuroscience Selected publications

Carli M, Kostoula C, Sacchetti G, Mainolfi P, Anastasia A, Villani C, Invernizzi RW. Tph2 gene deletion enhances amphetamine-induced hypermotility: effect of 5-HT restoration and role of striatal noradrenaline release. J Neurochem. 2015 Nov;135(4):674-85.

Carli M and Invernizzi RW. Serotoninergic and dopaminergic modulation of cortico-striatal circuit in executive and attention deficits induced by NMDA receptor hypofunction in the 5-choice serial reaction time task. Frontiers in Neural Circuits 8: 58. doi: 10.3389/fncir.2014.00058

Agnoli L, Mainolfi P, Invernizzi RW, Carli M.Dopamine D1-like and D2-like receptors in the dorsal striatum control different aspects of attentional performance in the five-choice serial reaction time task under a condition of increased activity of corticostriatal inputs.

Neuropsychopharmacology. 2013;38:701-14. Carli M, Calcagno E, Mainolfi P, Mainini E, Invernizzi RW. Effects of aripiprazole, olanzapine, and haloperidol in a model of cognitive

deficit of schizophrenia in rats: relationship with glutamate release in the medial prefrontal cortex. Psychopharmacology (Berl). 2011;214(3):639-52.

Carli M, Calcagno E, Mainini E, Arnt J, Invernizzi RW. Sertindole restores attentional performance and suppresses glutamate release induced by the NMDA receptor antagonist CPP. Psychopharmacology (Berl). 2011;214:625-37

Pozzi L, Sacchetti G, Agnoli L, Mainolfi P, Invernizzi RW, Carli M. Distinct Changes in CREB Phosphorylation in Frontal Cortex and Striatum During Contingent and Non-Contingent Performance of a Visual Attention Task. Front Behav Neurosci. 2011;5:65.

Pozzi L, Baviera M, Sacchetti G, Calcagno E, Balducci C, Invernizzi RW, Carli M. Attention deficit induced by blockade of N-methyl D-aspartate receptors in the prefrontal cortex is associated with enhanced glutamate release and cAMP response element binding protein phosphorylation: role of metabotropic glutamate receptors2/3. Neuroscience. 2011;176:336-48

Pozzi L, Greco B, Sacchetti G, Leoni G, Invernizzi RW, Carli M. Blockade of serotonin 2A receptors prevents PCP-induced attentional performance deficit and CREB phosphorylation in the dorsal striatum of DBA/2 mice. Psychopharmacology (Berl). 2010; 208:387-99.

Noe F, Vaghi V, Balducci C, Fitzsimons H, Bland R, Zardoni D, Sperk G, Carli M, During MJ, Vezzani A. Anticonvulsant effects and behavioural outcomes of rAAV serotype 1 vector-mediated neuropeptide Y overexpression in rat hippocampus. Gene Ther. 2010 17::643-52.

Calcagno E, Carli M, Baviera M, Invernizzi RW. Endogenous serotonin and serotonin2C receptors are involved in the ability of M100907 to suppress cortical glutamate release induced by NMDA receptor blockade. J Neurochem. 2009; 108:521-32.

Noè F, Frasca A, Balducci C, Carli M, Sperk G, Ferraguti F, Pitkonen A, Bland R, Fitzsimons H, During M, Vezzani A. Neuropeptide Y overexpression using recombinant adeno-associated viral vectors. Neurotherapeutics. 2009; 6:300-6.

Baviera M, Invernizzi RW, Carli M. Haloperidol and clozapine have dissociable effects in a model of attentional performance deficits induced by blockade of NMDA receptors in the mPFC. Psychopharmacology (Berl). 2008 196:269-80.

Dossena S, Imeri L, Mangieri M, Garofoli A, Ferrari L, Senatore A, Restelli E, Balducci C, Fiordaliso F, Salio M, Bianchi S, Fioriti L, Morbin M, Pincherle A, Marcon G, Villani F, Carli M, Tagliavini F, Forloni G, Chiesa R. Mutant prion protein expression causes motor and memory deficits and abnormal sleep patterns in a transgenic mouse model. Neuron. 2008; 60:598-609.

Luca Pasina obtained the doctor of Pharmacy degree at the University of Pavia during 2003 and is currently researcher at the Istituto di Ricerche Farmacologiche Mario Negri. The main research activity involve: pharmacoepidemiological study for the evaluation of appropriate drug use in the elderly, the risk of adverse drug reactions, drug-drug interactions and adverse effects on cognitive and physical functions; and the development and update of INTERCheckWeb, a Computerized Prescription Support System (CPSS) to support physicians during clinical practice. CPSS has been developed to improve the appropriateness of drug prescription; it sotres information about drug-drug interaction, potentially inappropriate medications, anticholinergic burden and dose adjustment in case of renal disease and modality for drug withdrawals (dose tapering). Research activities involve also studies to improve the use generic drugs and biosimilars. Selected publications

Pasina L, Zanotta D, Puricelli S, Djignefa DC, Bonoldi G. Proton pump inhibitors and risk of hypomagnesemia. Eur J Intern Med. 2015; 26:e25-6.

Pasina L, Brucato AL, Djade CD, Di Corato P, Ghidoni S, Tettamanti M, Franchi C, Salerno F, Corrao S, Marengoni A, Marcucci M, Mannucci PM, Nobili A; REPOSI Investigators. Inappropriate prescription of allopurinol and febuxostat and risk of adverse events in the elderly: results from the REPOSI registry. Eur J Clin Pharmacol. 2014; 70:1495-503.

Pasina L, Djade CD, Tettamanti M, Franchi C, Salerno F, Corrao S, Marengoni A, Marcucci M, Mannucci PM, Nobili A; REPOSI Investigators. Prevalence of potentially inappropriate medications and risk of adverse clinical outcome in a cohort of hospitalized elderly patients: results from the REPOSI Study. J Clin Pharm Ther. 2014; 39:511-5.

Pasina L, Brucato AL, Falcone C, Cucchi E, Bresciani A, Sottocorno M, Taddei GC, Casati M, Franchi C, Djade CD, Nobili A. Medication non-adherence among elderly patients newly discharged and receiving polypharmacy. Drugs Aging. 2014; 31:283-9.

Pasina L, Djade CD, Nobili A, Tettamanti M, Franchi C, Salerno F, Corrao S, Marengoni A, Iorio A, Marcucci M, Mannucci P. Drug-drug interactions in a cohort of hospitalized elderly patients. Pharmacoepidemiol Drug Saf. 2013; 22:1054-60.

Ghibelli S, Marengoni A, Djade CD, Nobili A, Tettamanti M, Franchi C, Caccia S, Giovarruscio F, Remuzzi A, Pasina L. Prevention of inappropriate prescribing in hospitalized older patients using a computerized prescription support system (INTERcheck(®)). Drugs Aging. 2013; 30:821-8.

Pasina L, Djade CD, Lucca U, Nobili A, Tettamanti M, Franchi C, Salerno F, anticholinergic burden with cognitive and functional status in a cohort of hospitalized elderly: comparison of the anticholinergic cognitive burden scale and anticholinergic risk scale: results from the REPOSI study. Drugs Aging. 2013;30:103-12.

Teresa Ravizza got her Doctoral Degree in Biological Sciences in 1996 at the University of Milano. Then she got a Master in “Research Specialist in Pharmacology” at Mario Negri Institute in 2000. She spent her post-doc training at the Albert Einstein College of Medicine of New York in 2000-2001, where she studied the mechanisms underlying epileptogenesis in experimental models of pediatric epilepsy. She spent additional post-doc periods at the Academic Medical Center of Amsterdam and at University of Irvine (UCI), California (USA) between 2005 and 2009. Since

2010, she is the head of the Unit of Pathophysiology of Neuron-Glia Communication. Her scientific interest is to characterize changes in the expression of molecules produced by astrocytes and microglia in various pathological conditions, such as epilepsy, trauma, excitotoxicity and inflammation. A special focus is given to the pro- and anti-inflammatory molecules, and to the role played by these mediators in mediating functional and biochemical alteration in the brain (neuronal cell loss, neuronal excitability, alteration in blood-brain barrier permeability). Selected pubblications

Ravizza T, Boer K, Redeker S, Spliet WGM, van Rijen PC, Troost D, Vezzani A, Aronica E. (2006) The IL-1 system in epilepsy-associated malformations of cortical development. Neurobiol Dis, 24, 128

Ravizza T, Lucas SM, Balosso S, Bernardino L, Ku G, Noè F, Malva J, Randle JC, Allan S, Vezzani A. (2006) Inactivation of caspase-1 in rodent brain: a novel anticonvulsive strategy. Epilepsia, 47, 1160

Ravizza T, Gagliardi B, Noé F, Boer K, Aronica E, Vezzani A. (2008) Innate and adaptive immunità during epiletogenesis and spontaneous seizures: evidence from experimental models and human temporal lobe epilepsy. Neurobiol Dis, 29, 142

Marcon J, Gagliardi B, Noé F, Morin M, Lerner-Natoli M, Vezzani A, Ravizza T. Age-dependent vascular changes induced by status epilepticus in rat forebrain: implication for epileptogenesis (2009) Neurobiol Dis, 34, 121

Maroso M, Balosso S, Ravizza T, Liu J, Aronica E, Iyer AM, Rossetti C, Molteni M, Casalgrandi M, Manfredi AA, Bianchi ME, Vezzani A. Toll-like receptor 4 (TLR4) and High Mobility Group Box 1 (HMGB1) are involved in ictogenesis and can be targeted to reduce seizures (2010) Nature Medicine, 16, 413

Dubé C, Ravizza T, Hamamura T, Zha Q, Keebaugh A, Fok K, Andres A, Nalcioglu O, Obenaus A, Vezzani, Baram TZ. Epileptogenesis provoked by prolonged experimental febrile seizures: mechanisms and biomarkers (2010) J Neurosci, 30, 7484

Ravizza T, Balosso S, Vezzani A Inflammation and prevention of epileptogenesis. Neurosci Lett. 2011; 497:223-30 Akin D, Ravizza T, Maroso M, Carcak N, Eryigit T, Vanzulli I, Aker RG, Vezzani A, Onat FY. IL-1β is induced in reactive astrocytes in

the somatosensory cortex of rats with genetic absence epilepsy at the onset of spike-and-wave discharges, and contributes to their occurrence. Neurobiol Dis. 2011; 44:259-69

Filibian M, Frasca A, Maggioni D, Micotti E, Vezzani A, Ravizza T. In vivo imaging of glia activation using 1H-magnetic resonance spectroscopy to detect putative biomarkers of tissue epileptogenicity. Epilepsia. 2012;53:1907-16.

Librizzi L, Noé F, Vezzani A, de Curtis M, Ravizza T. Seizure-induced brain-borne inflammation sustains seizure recurrence and blood-brain barrier damage. Ann Neurol. 2012; 72:82-90.

Noé FM, Polascheck N, Frigerio F, Bankstahl M, Ravizza T, Marchini S, Beltrame L, Banderó CR, Löscher W, Vezzani A. Pharmacological blockade of IL-1β/IL-1 receptor type 1 axis during epileptogenesis provides neuroprotection in two rat models of temporal lobe epilepsy. Neurobiol Dis. 2013 59:183-93.

Iori V, Maroso M, Rizzi M, Iyer AM, Vertemara R, Carli M, Agresti A, Antonelli A, Bianchi ME, Aronica E, Ravizza T, Vezzani A. Receptor for Advanced Glycation Endproducts is upregulated in temporal lobe epilepsy and contributes to experimental seizures. Neurobiol Dis. 2013;58:102-14.

Mishto M, Raza ML, de Biase D, Ravizza T, Vasuri F, Martucci M, Keller C, Bellavista E, Buchholz TJ, Kloetzel PM, Pession A, Vezzani A, Heinemann U. The immunoproteasome β5i subunit is a key contributor to ictogenesis in a rat model of chronic epilepsy. Brain Behav Immun. 2015; 49:188-96

Emma Riva, Medical Doctor degree in 1984 University of Milan, PhD in 1990 in Cardiovascular Pathophysiology at the University of London (UK) Training: Research Assistant, Department of Pharmacology, Medical School, University of Ottawa, Canada; Internship in Internal Medicine, Ospedale Luigi Sacco, Milan; Cardiac Fellow, St Thomas' Hospital, London, UK. Field of interest: Prevalence and effects of anemia on cognitive, functional and clinical variables in the elderly; Problem behaviors in dementia; Burden for care-givers of Alzheimer Disease patients; End of life care. Present and past roles in Institute Head of the Geriatric Pharmacology Unit, Istituto "Mario Negri", Milan; Scientific Director of the hospice “Via di Natale Franco Gallini”, Aviano, Italy; Consultant Istituto Geriatrico “Pio Albergo Trivulzio”, Milan: Project member of PREDICT (Policy Review and Evaluation of Dementia and Institutional Care Trends): a Transnational Comparison.

Selected publications

Avanzini F, Mafrici A, Riva E, Franzosi MG, Milani Va, Giudici V, Marelli G, Mariani G, Piatti PM, Roncaglioni MC on behalf of

GLICINE-SPIDER Collaborative Group A multicenter observational study on the management of hyperglycemia in patients with acute coronary syndrome Project. Nutr Metab Cardiovasc Dis 2015;25:916-925

Mandelli S, Riva E, Tettamanti M, Detoma P, Giacomin A, Lucca U. Reduced Estimated Glomerular Filtration Rate according to Five Different Equations and Risk of Mortality in the Oldest Old: The Health and Anemia Population-based Study. 2015 Plos One 10(8):e0136039.

Santalucia P, Baviera M, Cortesi L, Tettamanti M, Riva E, Roncaglioni MC. Epidemiological trends in stroke from 2002 to 2010: results from a large Italian population based study 2015 J Stroke Cerebrovasc Dis 2015 Jun 4. pii: S1052-3057(15)00253-0

Marelli G, Avanzini F, Iacuitti G, Planca E, Frigerio I, Busi G, Carlino L, Cortesi L, Roncaglioni MC, Riva E. Effectiveness of a nurse-managed protocol to prevent hypoglycemia in hospitalized patients with diabetes. J Diabetes Res 2015

Avanzini F, Marelli G, Saltafossi D, Longhi C, Carbone S, Carlino L, Planca, E Vilei V, Roncaglioni MC, Riva E. Effectivness safety and feasibility of an evidence-based insulin infusion protocol targeting moderate glycemic control in intensive cardiac care unit. Eur Heart J: Acute Cardiovascular Care 2015

Lucca U, Tettamanti M, Logroscino G, Tiraboschi P, Landi C, Sacco L, Garrì M, Ammesso S, Bertinotti C, Biotti A, Gargantini E, Piedicorcia A, Nobili A, Pasina L, Franchi C, Djade CD, Riva E, Recchia A. Prevalence of dementia in the oldest old: The Monzino 80-plus population based study. Alzheimer's & Dementia 2015;11:258-270

De LaHiguera L, Riva E, Djade CD, Mandelli S, Franchi C, Marengoni A, Salerno F, Corrao S, Pasina L, Tettamanti M, Marcucci M, Mannucci PM, Nobili A. Prognostic value of estimated glomerular filtration rate in hospitalized elderly patients. Intern Emerg Med 2014;9(7):735-47

Baviera M, Santalucia P, Cortesi L, Marzona I, Tettamanti M, Avanzini F, Nobili A, Riva E, Caso V, Fortino I, Bortolotti A, Merlino L, Roncaglioni MC. Sex differences in cardiovascular outcomes, pharmacological treatments and indicators of care in patients with newly diagnosed diabetes: analyses on administrative database. Eur J Intern Med 2014;25:270-275

L Monesi, M Tettamanti, L Cortesi, M Baviera, I Marzona, F Avanzini, G Monesi, A Nobili, E Riva, I Fortino, A Bortolotti, G Fontana, L Merlino, R Trevisan, MC Roncaglioni. Elevated risk of death and major cardiovascular events in subjects with newly diagnosed diabetes: study on administrative database. Nutrition, Metabolism & Cardiovascular Diseases 2014;24:263-270

Riva E, Coppa A, Tettamanti M, Lucca U, Garattini S, Gallini C, Marson R, Garattini S. Dieci anni di “esperienze” dell’hospice “via di Natale”. Rivista Italiana di Cure Palliative 2012;14(1):19-25

Franchi C, Tettamanti M, Marengoni A, Bonometti F, Pasina L, Cortesi L, Fortino I, Bortolotti A, Merlino L, Lucca U, Riva E, Nobili A. Changes in trend of antipsychotics prescription in patients treated with cholinesterase inhibitors after warnings from Italian Medicines Agency. Results from the EPIFARM-Elderly Project. Eur Neuropsychopharmacol. 2012; 22: 569-77

Monesi L, Baviera M, Marzona I, Avanzini F, Monesi G, Nobili A, Tettamanti M, Cortesi L, Riva E, Fortino I, Bortolotti A, Fontana G, Merlino L, Roncaglioni MC. Prevalence, incidence and mortality of diagnosed diabetes: evidence from an Italian population-based study: Diabet Med. 2012; 29:385-92

Franchi C, Lucca U, Tettamanti M, Riva E, Fortino I, Bortolotti A, Merlino L, Pasina L, Nobili A. Cholinesterasi inhibitor use in, in Alzheimer’s disease: the PEIFARM-Elderly Project. Pharmacoepidemiology and Drug Safety, 2011; 20:497-505

Nobil i A, Franchi C, Pasina L, Tettamanti M, Baviera M, Monesi L, Roncaglioni C, Riva E, Lucca U, Bortolotti A, Fortino, I Merlino L. Drug utilization and polypharmacy in an Italian elderly population: the EPIFARM-Elderly Project. Pharmacoepidemiology and Drug Safety, 2011; 20:488-96

Lucca U, Garrì MT, Recchia A, Logroscino G, Tiraboschi P, Franceschi M, Bertinotti C, Biotti A, Gargantini E, Maragna M, Nobili A, Pasina L, Franchi C, Riva E, Tettamanti M. A population-based study of dementia in the oldest old: The Monzino 80-plus. Study design, methodological challenges, and population characteristics. BMC Neurol. 2011; 25:11-4

Baviera M, Monesi L, Garzona I, Avanzino F, Monesi G, Nobili A, Tettamanti M, Riva E, Cortesi L, Bortolotti A, Fortino I, Merlino L, Fontana G, Roncaglioni MC. Trends in drug prescriptions to diabetic patients from 2000 to 2008 in Italy's Lombardy Region: a large population-based study. Diabetes Res Clin Pract. 2011; 93:123-30

Mauro Tettamanti got his Biology Degree at the Università degli Studi di Milano in 1986, and the specialisation in Epidemiology and Medical Statistics in 1993, at the Università degli Studi di Pavia. Teaching experience Introduction course to statistics, Master in Ergonomy, Politecnico di Milano, years 2001-2004 Areas of interest: Planning, conduction and analysis of clinical trials and epidemiologic researches in the geriatric field: Phase I, II, III and observational studies on the efficacy of drugs on neurologic disorders, with special emphasis on dementia; Effects of multi-disciplinary interventions on geriatric/dementia patients; Epidemiology and risk factors of dementia; Care of patients with terminal illness; Association of anemia with prevalence of diseases and cognitive problems Scholarship between 1989 and 1998, Senior Researcher since 1999 and Head of the Unit of Geriatric Epidemiology at the Mario Negri Institute since 2001. Selected publications

Lucca U, Nobili A, Riva E, Tettamanti M. Cholinesterase inhibitor use and age in the general population. Arch Neurol 2006; 63:154-155 Lucca U, Tettamanti M, Quadri P. Homocysteine lowering and cognitive performance. N Engl J Med 2006; 355:1390 Tettamanti M, Garri' M T, Nobili A, Riva E, Lucca U. Low folate and the risk of cognitive and functional deficits in the very old: The

Monzino 80-plus study. J Am Coll Nutr 2006; 25: 502-508 Tettamanti M, Lucca U, Gandini F, Recchia A, Mosconi P, Apolone G, et al. Prevalence, incidence and types of mild anemia in the

elderly: the "Health and Anemia" population-based study. Haematologica 2010; 95:1849-56 Nobili A, Franchi C, Pasina L, Tettamanti M, Baviera M, Monesi L, et al. Drug utilization and polypharmacy in an Italian elderly

population: the EPIFARM-elderly project. Pharmacoepidemiol Drug Saf 2011; 20:488-496. Parabiaghi A, Franchi C, Tettamanti M, Barbato A, D'Avanzo B, Fortino I, Bortolotti A, Merlino L, Nobili A. The declining use of

reboxetine in years 2000 to 2006: a pharmacoepidemiological comparative study.J Clin Psychopharmacol. 2012; 32:303-5 Haik S, Marcon G, Mallet A, Tettamanti M, Welaratne A, Giaccone G, et al. Doxycycline in Creutzfeldt-Jakob disease: a phase 2,

randomised, double-blind, placebo-controlled trial. Lancet Neurol 2014; 13:150-158 Lucca U, Tettamanti M, Logroscino G, Tiraboschi P, Landi C, Sacco L, Garri' M T, Ammesso S, Bertinotti C, Biotti A, Gargantini E,

Piedicorcia A, Nobili A, Pasina L, Franchi C, Djade C D, Riva E, Recchia A. Prevalence of dementia in the oldest old: The Monzino 80-plus population based study. Alzheimers Dement. 2015; 11:258-270

Pietro Veglianese got a degree in Chemistry and Pharmaceutical Technologies in 2000 at the University of Milan. In 2005 he got a degree as Pharmacological Research Specialist at the Department of Neuroscience (Laboratory of Molecular Neurobiology) Istituto di Ricerche Farmacologiche Mario Negri of Milan. In 2007 he got his PhD at the Open University (UK). Since January 2014 he is Head of Acute Spinal Cord Injury and Neuroregeneration unit at the Istituto di Ricerche Farmacologiche Mario Negri of Milan. His research interests involves the development and characterization of biomaterials able to deliver drugs in situ for treating spinal cord injury. His works have shown that the use of biomaterials, namely hydrogels and nanoparticles, have the potential to release and increase the effectiveness of different drugs, paving the base for a new delivery therapeutic strategy. In addition, some smart delivery tools (nanoparticles) developed by our group are able to target selectively drugs in specific cells of the central nervous system (microglia / macrophages), opening a new concept of therapy in the treatment of inflammation (cell-targeted therapy). With a bioengineering approach, the unit is also pursuing a project involving the development of a combined therapy of different drugs and stem cells (synergistic effect) administered at the site of injury through biomaterials. These projects are carried out with a multidisciplinary collaboration that involves Politecnico di Milano (biomaterials), SUPSI Lugano

(biomaterials) and Cell Factory of the Policlinico of Milan (stem cells). In addition, Pietro Veglianese was referee for several scientific journals (Brain, Journal of Controlled Release, Cell Biology and Toxicology, ACS Nano and British Journal of Pharmaceutical Research) Selected publications:

Papa, S., R. Ferrari, M. De Paola, F. Rossi, A. Mariani, I. Caron, E. Sammali, M. Peviani, V. Dell'oro, C. Colombo, M. Morbidelli, G. Forloni, G. Perale, D. Moscatelli and P. Veglianese (2013). "Polymeric nanoparticle system to target activated microglia/macrophages in spinal cord injury." J Control Release 174C: 15-26.

Papa, S., F. Rossi, R. Ferrari, A. Mariani, M. De Paola, I. Caron, F. Fiordaliso, C. Bisighini, E. Sammali, C. Colombo, M. Gobbi, M. Canovi, J. Lucchetti, M. Peviani, M. Morbidelli, G. Forloni, G. Perale, D. Moscatelli and P. Veglianese (2013). "Selective nanovector mediated treatment of activated proinflammatory microglia/macrophages in spinal cord injury." ACS Nano 7(11): 9881-9895.

Rossi, F., G. Perale, S. Papa, G. Forloni and P. Veglianese (2013). "Current options for drug delivery to the spinal cord." Expert Opin Drug Deliv 10(3): 385-396.

Perale, G., F. Rossi, M. Santoro, M. Peviani, S. Papa, D. Llupi, P. Torriani, E. Micotti, S. Previdi, L. Cervo, E. Sundstrom, A. R. Boccaccini, M. Masi, G. Forloni and P. Veglianese (2012). "Multiple drug delivery hydrogel system for spinal cord injury repair strategies." J Control Release 159(2): 271-280.

Perale, G., F. Rossi, E. Sundstrom, S. Bacchiega, M. Masi, G. Forloni and P. Veglianese (2011). "Hydrogels in spinal cord injury repair strategies." ACS Chem Neurosci 2(7): 336-345.

Elisa R Zanier. 1998, Medical Doctor degree (110/110) at the University of Milano, Italy. 1998/2001: Residency in Anesthesiology and Critical Care Medicine at the University of Milano. 2 years Post-doctoral fellowship at the Neurotrauma Laboratory-Neurosurgery Division, University of Los Angeles, California (UCLA), USA. 2003-2008 Assistant physician in the Neurosurgical Intensive Care Unit, Department of Anesthesia and Critical Care Medicine, Fondazione IRCCS Ospedale Maggiore Policlinico, Milano. Since 2007: Teaching assignment into postgraduate school of Critical Care Medicine and Anesthesiology, University of Milano. Since 2008: Associate researcher at the Laboratory of Inflammation and Nervous System Diseases, Mario Negri Institute. Since 2012 Head of the Unit of Cell Therapy and Acute Brain Injury, Mario Negri Institute, Milano. Present interests include: experimental models: traumatic brain injury and stroke. 2013: Fulfillment of the criteria for Associate Professor in Anesthesia and Critical Care Medicine (national certification). 2014 received the Rita Levi Montalcini prize from the European Society of Intensive Therapy, editor of Intensive Care Medicine Experimental. Scientific fields: pathophysiology of traumatic brain injury and brain ischemia; brain repair; inflammation as target of therapeutic strategies; the protective mechanisms of stem cells. Publications in PubMed: 47. H index 19 Selected publications

Zanier ER, Marchesi F, Ortolano F, Perego C, Arabian M, Zoerle T, Sammali E, Pischiutta F, De Simoni MG. Fractalkine receptor deficiency is associated with early protection but late worsening of outcome following brain trauma in mice. J Neurotrauma Sep 8, 2015. [Epub ahead of print].

Stocchetti N, Taccone FS, Citerio G, Pepe PE, Le Roux PD, Oddo M, Polderman KH, Stevens RD, Barsan W, Maas AI, Meyfroidt G, Bell MJ, Silbergleit R, Vespa PM, Faden AI, Helbok R, Tisherman S, Zanier ER, Valenzuela T, Wendon J, Menon DK, Vincent JL. Neuroprotection in acute brain injury: an up-to-date review. Crit Care 19; 186, 2015.

Zanier ER, Pischiutta F, Riganti L, Marchesi F, Turola E, Fumagalli S, Perego C, Parotto E, Vinci P, Veglianese P, D'Amico G, Verderio C, De Simoni MG. Neurotherapeutics 11; 679-95, 2014.

Zanier ER, Zangari R, Munthe-Fog L, Hein E, Zoerle T, Conte V, Orsini F, Tettamanti M, Stocchetti N, Garred P, De Simoni MG. Neurology 82; 126-34, 2014.

Pischiutta F, D'Amico G, Dander E, Biondi A, Biagi E, Citerio G, De Simoni MG, Zanier ER. Immunosuppression does not affect human bone marrow mesenchymal stromal cell efficacy after transplantation in traumatized mice brain. Neuropharmacology 79; 119-26, 2014.

Zanier ER, Montinaro M, Viganò M, Villa P, Fumagalli S, Pischiutta F, Longhi L, Leoni ML, Rebulla P, Stocchetti N, Lazzari L, De Simoni MG. Human umbilical cord blood mesenchymal stem cells protect mice brain after trauma. Crit Care Med. 39; 2501-2510, 2011.

Zanier ER, Brandi G, Peri G, Longhi L, ZoerleT, Tettamanti M, Garlanda C, Sigurtà A, Valaperta S, Mantovani A, De Simoni MG, Stocchetti N. Cerebrospinal fluid pentraxin 3 early after subarachnoid hemorrhage is associated with vasospasm. Intensive Care Med. 37; 302-9, 2011.

Stocchetti N, Colombo A, Ortolano F, Videtta W, Marchesi R, Longhi L, Zanier ER. Time course of intracranial hypertension after traumatic brain injury. J Neurotrauma 24; 1339-46, 2007.

Zanier ER, Ortolano F, Ghisoni L, Losappio S, Colombo A, Stocchetti N. Intracranial pressure monitoring in intensive care: clinical advantages of a computerized system over manual recording. Crit Care 11; R7, 2007.

Stocchetti N, Protti A, Lattuada M, Magnoni S, Longhi L, Ghisoni L, Egidi M, Zanier ER: Impact of pyrexia on neurochemistry and cerebral oxygenation after acute brain injury. J Neurol Neurosurg Psychiatry 76; 1135-1139, 2005.

Longhi L, Zanier ER, Royo N, Stocchetti N, McIntosh TK. Stem cell transplantation as a therapeutic strategy for traumatic brain injury. Transplant Immunology 15; 143–148, 2005.

Zanier ER, Lee S, Vespa P, Giza C, Hovda D: Increased hippocampal CA3 vulnerability to low-level kainic acid following lateral fluid percussion injury. J Neurotrauma 20; 409-420, 2003. Ip EY, Zanier ER, Moore AH, Lee SM, Hovda DA: Metabolic, Neurochemical, and Histologic

ACTIVITIES

The Department of Neuroscience is formed by eleven Laboratories; the activities of research are dedicated to the study

of neurological and psychiatric diseases, evaluated by the biological point of view, clinical and epidemiological aspects and the quality of care. In the Department, activities like drug information service, preparation of clinical trial protocols and epidemiological studies are developed not only in the neurological field. Traditionally, part of the Department is devoted to the development of experimental models for the pharmacological, neurochemical and pathogenetic studies in Alzheimer, Parkinson or prion's diseases, epilepsy, depression and cognitive impairment. More recently, consolidated expertise were established in the studies of amyotrophic lateral sclerosis (ALS) pathogenesis, cerebral stroke, trauma and drug abuse. Some of these disorders, like epilepsy, ALS and Alzheimer's disease, are investigated from the clinical and epidemiological points of view to evaluate drug and care efficacy. Genetic investigations and the use of biomaterial tools are the last acquired expertise. The activities of the Department are aimed to the integration of the different expertise to develop multidisciplinary approaches. The purpose is to address at different levels, knowledge, therapy and clinical practice to the numerous questions, largely unresolved, proposed by the disorders of nervous system.

MAIN FINDINGS The treatment with functionalized liposomes with ApoE peptide to facilitate the blood brain barrier passage and phosphatidic acid with anti-amyloidogenic activity, in murine models od Alzheimer’s disease reduces the brain amyloid burden and the presencve of amyloid oligomers with positive consequence on cognitive behavior. The intracerebral application of synthetic amyloid 1-40 e 1-42 in oligomeric form is associated with a cognitive damage, the effect is partially due to inflammatory mechanism mediated by non-neuronal cells. The cogntive damage induced by -synuclein, essential component of intracellular aggregates, named Lewy bodies, found in Parkinson disease brain and in other neurodegenerative disorders, injected in oligomeric form intracerebraventricularly, is associated with a glial activation distiguishable from the activation induced by amyloid oligomers The treatment with doxycycline non only antagonizes the toxic effects of amyloid oligomers but also the cognitive decline induced by LPS showing an anti-inflammatory component. An A peptide with transmembrane sequence TAT (TAT 1-6 A2V) including the mutation that in homozygosis is associated to the Alzhiemr’s disease, antagonized the toxic effect induced by amyloid in vitro and in vivo models The new peptide GADD45β-I (growth arrest and DNA damage-inducible 45β) inhibitor of MKK7 (mitogen-activated protein kinase kinase 7) protects from ischemic damage in two murine models with an interesting therapeutic window. We have identified new compounds that have the advantage of combining an anti-PrPSc activity with the ability to decrease PrPC level. These are being tested in cell and mouse models of acquired and genetic prion disease. The investigations in transgenic mice showing that the main cause of synaptic dysfunction in genetic prion's disorders is the accumulation of mutated PrP within the neuronal secretory pathway It has been established a clinical protocol to evaluate the efficacy of doxycycline in subjects with genetic risk to develop fatal insomnia (FFI), a genetic form of prion's disease. The incidence of the disease will be evaluated in subjects carryng the prion D178N mutation and treated with doxycycline for ten years. Evidence has been reported linking HSV1 to Alzheimer's disease. In a prospective population-based study in the oldest old (Monzino 80-plus Study), Frequency of herpes labialis infection was lower among subjects with dementia at first visit than among those without. This difference remained significant when corrected for age, sex, and education and also when the multivariable model was further adjusted for other clinical conditions.

Cognitive impairment and dementia have been related to an increased risk for falls in older adults. In the same prospective population-based study (Monzino 80-plus Study), the occurrence of falls was not significantly associated with dementia. The association remained non-significant when possible confounding variables were entered as covariates into a multivariable model relating dementia to falls In the Centenari a Trieste Project approximately 50% of the population residing in Trieste and born in or before 1913 was recruited. Centenarians were evaluated from a clinical point of view by a physician and subsequently by neuropsychologists relative to cognition, mood and daily living activities. Subjects with good/very good results were tested with further neuropsychological instruments. Data for those who died before being assessed were acquired through administrative databases with the collaboration of the Local Health Unit. During 2015 the randomization of mild-to-moderate AD patients to the EU-FP7 NILVAD Study was completed. At the end of March 2015, 510 patients had been randomized all over Europe, 55 of whom in Italy. In the Health and Anemia population-based study in the oldest old, a moderate renal impairment (defined as a creatinine concentration ≥ 1.3 mg/dL in women and ≥ 1.5 mg/dL in men was associated with a worse cognitive performance on the Mini-Mental State Examination test. Reduced kidney function could thus play a role in the complex, multifactorial aging-related cognitive decline. Patients with dementia resident in Alzheimer’s special care units (ASCU) had a lower rate of hospitalisation and use of physical restraints than those in traditional nursing homes In ASCU 60% of patients with dementia were taking at least one antipsychotic, 49% typical and 51% atypical. More than 50% of patients exposed to antipsychotics at baseline, were still taking the drug after 18 months of follow-up. The use of antipsychotic agents was strongly related to the presence of agitation, irritability, delusions, anxiety, night-time behaviour and aberrant motor behaviour In the Lecco Local Health Authority 16% of elderly patients were exposed to potential severe drug-drug interactions; age and number of chronic drugs were associated with an increasing risk of DDIs. Since physicians still have some difficulty in managing this topic, it is essential to provide them with adequate information on which factors raise the risk of DDIs. Age, local health unit (LHU) of residence, number of drugs and co-prescribed PIDs were predictors of hospitalization for hemorrhage. During 2005 in Lombardy Region, 76% of the elderly aged 65 years ore more (76% women and 75% men) received at least one chronic drug, 46% were exposed to polypharmacy (46% women and 45% men) and 20% to chronic polypharmacy (18% women and 22% men). Elderly in the age groups of 75-79, 80-84 and 85-89 years had the highest risk to be exposed to chronic polypharmacy (OR 2.25; 95%CI: 2.23-2.27, OR 2.68; 95%CI: 2.65-2.71, and OR 2.84; 95%CI: 2.79-2.89 respectively). During 2005, 34 % of the population living in Lombardy Region received at least one antibiotic drug prescription. The highest prescription prevalence was observed in the 0-17 and 80 or more year age ranges (41.6% and 41.9%, respectively). Patients aged <18 years (OR= 1.73; 95% CI 1.73, 1.74), aged 65 or older (OR= 1.64; 95% CI 1.63, 1.65), and those that live in Brescia (OR 1.66, 95% CI 1.65, 1.66) had a statistically significant higher risk of antibiotic drug exposure. In a large population sample of subject living in Lombardy Region, the use of paroxetine and fluoxetine peaked in 2002 and then decreased. The prescripition rates of mirtazapine gradually increased all through the study period: from 0.07% in 2000 to 0.13% in 2006. On the contrary, the prescription rates of reboxetine showed a different trend and progressively decreased from 0.20 in 2000 to 0.04 in 2006. In a sample of 38 internal medicine and geriatric wards, at hospital admission 52% of 1332 elderly patients aged 65 years or older taken five or more different drugs (polypharmacy) and were in the ward for a mean of

11 days. At hospital discharge there was an increase in the rate of patient with polypharamacy (+13%) and with multiple disease (+16%). Among elderly patients admitted with a diagnosis of AFF to internal medicine wards, an appropriate antithrombotic prophylaxis was taken by less than 50%, with an underuse of VKAs prescription independently of the level of cardio-embolic risk. Hospitalization did not improve the adherence to guidelines.

‐ After multiadjustment, the diagnosis of dementia was associated with in-hospital death (OR = 2.1; 95% CI = 1.0 - 4.5). Having dementia and at least one adverse clinical event during hospitalization showed an additive effect on in-hospital mortality (OR = 20.7 ;95% CI = 6.9 – 61.9).

The strongest association between clusters of diseases and polypharmacy was found for diabetes mellitus plus CHD plus CVD, diabetes plus CHD, and HF plus atrial fibrillation (AF).

The prescription of typical antipsychotics has been associated with an increased risk of CVEs. After stratification, persons prescribed with AChEI did not show any association with CVEs.

Nineteen percent of patients admitted to internal medicine and geriatric hospital wards are re-hospitalized at least once within 3 month after discharge. Adverse events during hospitalization, previous hospital admission, and vascular and liver diseases were significantly associated with likelihood of readmission.

We found a significant association with an increased risk of mortality at 3 months follow in patients exposed to at least 2 potentially severe DDIs (OR=2.62; 95% CI, 1.00-6.68; p=0.05). Hospitalization was associated to an increase in potentially severe DDIs. Careful monitoring for potentially severe DDIs, especially for those created at discharge or recently generated, is important to minimize the risk of associated harm.

We found that there were geographical differences in the prevalence of elderly people with chronic polypharmacy, only partly explained by health indicators. These findings highlight the need for targeted efforts on prescription practice to reduce polypharmacy.

In elderly hospitalized, severely reduced eGFR at the time of admission was associated with in-hospital mortality (OR 3.00; 95 % CI 1.20-7.39, p = 0.0230), but not with re-hospitalization (OR 0.97; 95 % CI 0.54-1.76, p = 0.9156) or mortality at 3 months after discharge (OR 1.93; 95 % CI 0.92-4.04, p = 0.1582). On the contrary, an increased risk (OR 2.60; 95 % CI 1.13-5.98, p = 0.0813) to die within 3 months after discharge was associated with decreased eGFR measured at the time of discharge.

REPOSI patients represent a population at high cardio-embolic and bleeding risks: most of them were at high cardio-embolic/high-intermediate bleeding risk (70.5% combining CHADS2 and HEMORR2HAGES, 98.3% combining CHA2DS2-VASc and HAS-BLED), and 50-60% of patients were classified in a cardio-embolic risk category higher than the bleeding risk category. In univariate and multivariable analyses, a higher bleeding score was negatively associated with warfarin prescription, and positively associated with aspirin prescription. The cardio-embolic scores were associated with the therapeutic choice only after adjusting for bleeding score or age.

Drugs with anticholinergic properties identified by the ACB scale and ARS are associated with worse cognitive and functional performance in elderly patients. The ACB scale might permit a rapid identification of drugs potentially associated with cognitive impairment in a dose-response pattern, but the ARS is better at rating activities of daily living.

The use of INTERCheck(®) was associated with a significant reduction in PIMs and new-onset potentially severe DDIs. CPSSs combining different prescribing quality measures should be considered as an important strategy for optimizing medication prescription for elderly patients.

The increasing number of drugs prescribed at hospital discharge is correlated to non-adherence and a high percentage of patients did not understand the purpose of their medications. Simplification of drug regimens and reduction of pill burdens should be targets for intervention.

Prevalence of inappropriate prescription of allopurinol remained almost the same at admission and discharge. Inappropriate use of this drugs is principally related to asymptomatic hyperuricemia. Careful assessment of clinical conditions and stricter adherence to evidence-based guidelines are essential for a rational use.

The documentation of delirium is poor in medical wards of Italian acute hospitals. Delirium was coded in 2.9%, while deficits in attention, orientation, and memory were found in 35.4%, 29.7% and 77.5% of patients. Neurocognitive deficits on objective testing (in a pattern suggestive of undiagnosed delirium) should be used to raise awareness of delirium, given their association with in-hospital mortality. The conservative attitude of Italian physicians to prescribe opioids in elderly patients changed very little between hospital admission and discharge through a period of 5 years. Reasons for such a low opioid prescription should be sought in physicians' and patients' concerns and prejudices.

Prevalence of inappropriate prescription of allopurinol remained almost the same at admission and discharge. Inappropriate use of this drugs is principally related to asymptomatic hyperuricemia. Careful assessment of clinical conditions and stricter adherence to evidence-based guidelines are essential for a rational use.

There was no evidence to consider couple therapy in the short-term as more or less effective than individual psychotherapy. The absence of a significant difference also held true when only distressed couples were included. However, in comparison to no/minimal treatment, the outcome for depressive symptoms and persistence of depression was better for couple therapy. There were no significant differences in the dropout rates between couple therapy and the individual psychotherapy, even when only distressed couples were analysed. Relationship distress was significantly lower and persistence of distress significantly less frequent in couple therapy than in the individual psychotherapy. This effect was enhanced when distressed couples were considered separately. The meta-synthesis of qualitative studies about frailty in the older populationshowed moderate trust of the stakeholders in screening and prevention of frailty. Whereas older people used practical and psychological strategies to resist frailty, these strategies were often overlooked by professionals. There is a direct correlation between ALS and mechanical trauma as a result of the following observations: The risk of ALS increases with the number of traumatic events and the severity of injuries. There is an inverse correlation between ALS and coffee intake. The prevalence of extrapyramidal signs in patients with ALS is higher than that expected in the general population. Early onset differs from late onset ALS for the higher exposure to lead, solvents, electromagnetic fields, and professional physical activity. There is an inverse correlation between physical exercise and ALS. However, among affected individuals the disease tends to occur at a younger age is the patient practiced physical exercise. Data on the 10-year mortality of ALS show a 12% survival rate with significant differences according to the phenotype at diagnosis. Predictors of long-term survival include younger age, possible/suspected ALS, spinal onset, and disease duration longer than 12 months. Survival of a male patient diagnosed at 75 years or older overlaps that of the general population. The diversity of ALS phenotype, incidence and outcome of ALS can be explained by the ethnic origin of the patients We have produced further evidence in support of the hypothesis that the speed of the disease depends not so much on the loss of motor neurons, but rather to a greater dysfunction of peripheral nerves leading to an earlier muscle denervation. This emphasizes that the ALS is a multysistem disorder and therefore in order to obtain a real therapeutic benefit is important to consider not only the protection of motor neurons but also to target other districts involved in the disease like the nerves, muscles and immune system. An immune response in the peripheral nervous system mediated by MHCI, CCL2 and complement C3 which seems aimed at preserving motor neuron This makes possible new direction for understanding the mechanisms of interaction between motor neurons and immune system and therefore highlight new therapeutic targets. For the first time a potential involvement of the chemokine CXCL13 in the pathogenesis and rapid progression of the disease.

A comprehensive rehabilitation program does not reduce the risk of falls in Parkinson disease when compared to usual care. In patients with epilepsy, an active monitoring of adverse events and drug interactions reduce significantly these events without addictive monetary costs. Epilepsy impairs all aspects of quality of life, although at different degree, both in children/adolescents and in their families. Parental apprehensiveness appears to have a role on this, and it may not reflect the severity of the disease. Administrative data are a cost-effective instrument to monitor epilepsy frequency. 1/6 of patients with active epilepsy in the general population have drug-resistant epilepsy and about 1/10 patients with newly diagnosed epilepsy will develop drug-resistant epilepsy. The long-term prognosis of epilepsy is favorable in most cases. Early seizure remission is not invariably followed by terminal remission and seizure outcome varies according to well-defined patterns. Prolonged seizure remission and terminal remission can be observed in patients with drug-resistant epilepsy suggesting that drug resistance is a dynamic process and, as such, can be reversible. Treatment of the first unprovoked seizure reduces the risk of a subsequent seizure but does not affect long-term seizure control. Antiepileptic drugs are associated with adverse events, but do not seem to affect the mortality rate. The decision to start antiepileptic drug treatment following a first unprovoked seizure should be individualized and based on patient preference, clinical, legal, and socio-cultural factors. In about one third of cases a confident diagnosis of PNES/ES can be established on clinical ground based on video data alone. This finding benefit all affected patients, particularly those with no access to video-EEG monitoring units. The Website can be successfully used by doctors working in different health systems. Concordance between neurologists assessing identical case scenarios for the diagnosis of epilepsy is moderate when based on history alone. In contrast, concordance on seizure type and etiology is poor. Concordance improves when results of investigations are included. The long-term remission of epilepsy occurs in almost one half of newly diagnosed patients and can be predicted on the basis of the clinical features of the subject. The predominance of a relapsing-remitting pattern suggests a dynamic course of the natural history of the disease, which is affected by drugs and requires a careful selection of patients for treatment discontinuation.

The crucial involvement of specific pro-inflammatory cytokines in seizure mechanisms using rodent models of epilepsy, thus describing a new pathological mechanism, i.e. neuroinflammation, which may be relevant for human epilepsy. This discovery highlights novel targets for developing anti-epiletogenic and disease modifying therapies translatable to the clinical setting.

In epilepsy animal models that astrocitic activation as assessed by MR spectroscopy can be used as a biomarker of epileptogenesis. This approach can be easily translated to the clinical setting for further validation in humans.

The membrane-bound drug transport efflux proteins are functionally activated by seizures and have a significant role in decreasing the brain concentrations of antiepileptic drugs in experimental models. Pharmacological intervention to block the activity of these proteins may contribute to reverse multidrug resistance in epilepsy.

The complement system is a relevant target in acute brain injury:

Recombinant complement inhibitor (rhC1-INH) has a powerful neuroprotective action and a wide therapeutic window in brain ischemia/reperfusion injury

Targeting mannose-binding lectin (MBL), an activator of the lectin complement pathway, leads to neuroprotection with a wide therapeutic window

In subarachnoid hemorrage (SAH) patients ficolin-3, one of the activators of the lectin complement pathway is associated to clinical and structural parameters of severity.

The lectin complement pathway initiators ficolins are consumed within 6h after stroke implicating activation of this pathway. Early ficolin-1 is selectively related to 3- months unfavourable outcome.

Microglia is associated to protective actions in the injured brain and mesenchymal stem cells drive protective microglia polarization in in vitro and in vivo injury.

Long term efficacy of human bone marrow mesenchymal stem cells in traumatized mice brain is not affected by immunosuppressive treatment, amniotic stem cells and their derivatives protect the injured brain. The deletion of Tph2, the gene responsible for the synthesis of brain serotonin, enhances the motor effect of amphetamine in mice through the inhibition of striatal NA release.

Patients affected by Rett syndrome may have alteration of brain cholesterol metabolism A single session of cocaine self-administration is sufficient to shape rat behaviour towards goal-directed behaviours and selectively up-regulate Arc expression in mPFC. This is the first evidence that the mPFC's function is already profoundly influenced by the first voluntary cocaine exposure. The use and the early phases of cocaine abstinence induce a finely tuned modulation of BDNF expression in the NAc and in the mPFC. Short abstinence from contingent cocaine i.v. self-administration elevates αCaMKII autophosphorilation in NAc and mPFC. The persistent enhancement in the mPFC of abstinent rats may represent a previously unappreciated contribution to initial incubation of cocaine-seeking. Environmental stimuli associated to drug self-administration induce drug-seeking behaviour when presented to rodents after a long period of abstinence. Bifeprunox, a partial agonist at DA D2 and 5-HT1A receptors, influences nicotine-seeking behaviour in response to drug-associated stimuli in rats. GlyT1-inhibitors might offer a therapeutic opportunity for acute cue-controlled nicotine-seeking. The lack of persistent effects of the sub-chronic treatment associated with nicotine cues exposure suggests that short-term administration of GlyT1-inhibitor SSR504734 is not sufficient to promote extinction of nicotine-cue conditioned responding.

NATIONAL COLLABORATIONS

Area di Neuroscienze, SISSA, Trieste Associazione Familiari Insonnia Familiare Fatale malattie da prioni, Treviso Associazione Mondiale di Riabilitazione Psicosociale, Sezione Italiana, Milano Associazione per la Ricerca Neurogenetica, Lamezia Terme (CS) e ASL 6, Regione Calabria Azienda Ospedaliera Ospedali Riuniti di Bergamo Azienda per l’Assistenza Sanitaria n.1, Triestina Bracco Imaging, Milano Cell Factory, Fondazione IRCCS Ospedale Maggiore Policlinico, Milano CEND, Centro Eccellenza per le Malattie Neurodegenerative, Università di Milano Centro di Ricerca “E. Menni” Poliambulanza, Brescia, Italy Centro di Terapie per l’Adolescenza, Milano Centro Medicina dell’Invecchiamento del Policlinico A. Gemelli, Università Cattolica a Roma Centro di Neurofarmacologia, Dipartimento di Scienze Farmacologiche, Università di Milano

Centro Nemo, Ospedale di Niguarda, Milano Centro Parkinson-Istituti Clinici di Perfezionamento Centro Studi sulla Sofferenza Urbana, SOUQ, Milano Clinica IRCSS S. Maria Nascente, Milano Clinica Neurologica, Università di Parma Clinica Psichiatrica, Università di Pavia Dipartimento di Salute Mentale, Azienda Provinciale Clinica Psichiatrica, Università di L’Aquila Clinica Psichiatrica Università degli Studi di Genova Consorzio MIA, Milano Department of Management, Economics and Industrial Engineering, Politecnico di Milano Department of Multi Criteria Decision Analysis (MCDA) for Decision Making, Politecnico di Milano Department of Quantitative Methods for Business Economic Sciences, Facoltà di Statistica, Università Bicocca, Milano DIBIT, San Raffaele Scientific Insitute, Milano Dipartimento di Anestesia e Rianimazione, Universià dif Milano, Fondazione IRCCS Ospedale Maggiore Policlinico, Milano Dipartimento di Biologia Funzionale e Strutturale Università dell’Insubria Dipartimento di Chimica Biologica, Università di Padova Dipartimento di Chimica, Università degli Studi di Milano Dipartimento di Chimica, Materiali e Ingegneria Chimica "Giulio Natta", Politecnico di Milano Dipartimento Dipendenze ASL di Como Dipartimento Endocrinologia, Università di Milano Dipartimento Farmaco Chimico Tecnologico, Università di Siena Dipartimento di Fisiologia Umana, Facoltà di Medicina, Università di Milano Dipartimento di Ingegneria e Scienza dell’Informazione, Università di Trento Dipartimento di Immunologia, Fondazione Humanitas per la Ricerca di Milano Dipartimento di Neurologia, Seconda Università di Napoli Dipartimento Neurologia, IRCCS Fondazione Maugeri, Pavia Dipartimento Neurologia, Ospedale Molinette, Torino Dipartimento di Neurologia Università di Milano, Ospedale Luigi Sacco. Dipartimento di Neuroscienze, Università di Parma, Parma Dipartimento di Neuroscienze e Organi di Senso, Università di Bari, Bari. Dipartimento di Neuroscienze, Oftalmologia e Genetica, Unità di Neuroimmunologia,Università di Genova Dipartimento di Salute Mentale di Azienda Ospedaliera di Lecco Dipartimento di Salute Mentale di Azienda Ospedaliera Sacco, Milano Dipartimento di Salute Mentale, Azienda Provinciale per i Servizi Sanitari di Trento, Trento Dipartimento di Salute Mentale di Grosseto Dipartimento di Salute Mentale di Niguarda, Milano Dipartimento di Scienze Biomediche e Cliniche Università di Milano, Milano Dip. di Scienze Biomolecolari e Biotecnologie, Università di Milano Dipartimento Scienze Cliniche e Sperimentali, Università degli Studi di Brescia Dipartimento di Scienze Cliniche e di Comunità, Università degli Studi di Milano Dipartimento di Scienze della Salute, Facoltà di Medicina e Chirurgia, Università degli Studi di Milano Dipartimento di Scienze della Salute, Università Milano- Bicocca, Monza Dipartimento di Scienze Fisiologiche Università di Pavia, Pavia Dipartimento di Scienze mediche e biologiche - Università di Udine Dipartimento Scienze Neurologiche, Università di Genova, Genova Dipartimento Scienze Neurologiche, Ospedale Maggiore Policlinico di Milano Direzione Generale Sanità, Regione Lombardia, Milano Divisione di Ematologia, Università di Pavia Fondazione IRCCS Policlinico S. Matteo, Pavia Federazione Alzheimer Italia, Milano Federazione delle Associazioni dei Dirigenti Ospedalieri Internisti, FADOI Federazione Italiana dei Medici di Medicina Generale Franco Calori Cell Factory, Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano Fondazione Cenci-Gallingani, Abbiategrasso, MI,.

Fondazione IRCCS Istituto Neurologico Carlo Besta, Milano Hospice “Franco Gallini”, Aviano (PN) IRCSS "Casa Sollievo della Sofferenza", San Giovanni Rotondo Istituto di Ricovero e Cura a Carattere Scientifico IRCCS (I.N.R.C.A.), Ancona IRCCS “Eugenio Medea” - Polo di Conegliano - Pieve di Soligo (TV) IRCSS Fatebenefratelli di Brescia IRCCS Ospedale Maggiore Policlinico, Milano IRCSS "San Raffaele", Milano Istituto di Farmacologia, Università di Milano Istituto di Fisiologia Umana II Università degli Studi di Milano, Milano Istituto Italiano di Tecnologia, Genova Istituto Neurologico "Casimiro Mondino", Pavia Istituto Scientifico Humanitas Istituto di Scienze e Tecnologie della Cognizione, CNR, Roma Istituto "Stella Maris", IRCCS, Calambrone (PI) Istituto Superiore di Sanità, Roma Istituto Zooprofilattico Piemonte Liguria Val D'Aosta,Torino Laboratorio di Epidemiologia e Neuroimaging e U.O. Alzheimer, IRCCS Fatebenefratelli, Brescia. Laboratory for Cell Therapy “Stefano Verri”, Paediatric Department, University of Milano-Bicocca, San Gerardo Hospital, Monza, Italy Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, Genova Nico, Neuroscience Institute Cavalieri Ottolenghi, Torino Ospedale del Bambin Gesu’, Roma Ospedale Regionale Ca Fondello, Treviso Ospedale "Molinette", Torino Ospedale “San Gerardo”, Monza Polo Tecnologico, IRCCS S. Maria Nascente, Fondazione Don Carlo Gnocchi Onlus, Milano Scuola Normale Superiore, Laboratorio NEST: National Enterprise nanoScience and nanoTechnology, Pisa Scuola di Specializzazione in Terapia Cognitiva “Studi Cognitivi”, Milano Servizio di Epidemiologia e Laboratorio di Analisi, ASL BI, Biella Società Italiana Medicina Interna, Roma Terzo Dipartimento di Medicina Interna, Medicine Operative, Unità Oderzo − ASL 9 Treviso Unità di Geriatria, Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico Unità di Neurologia, Opsedale San Gerardo di Monza. Unità Operativa di Neurologia, Casa di Cura S. Maria (Multimedica), Castellanza (VA). Unità Operativa di Neurologia Riabilitativa , Centro S. Maria Nascente, Fondazione Don Carlo Gnocchi Onlus, Milano. Unità di Patologia Umana e Istologia, Dip. Scienze mediche di Base, Università di Bari Unità di Patologia e Medicina Orale, Dipartimento di Scienze Chirurgiche Ricostruttive e Diagnostiche, Università degli Studi di Milano Unità di Statistica Medica, Biometria and Bioinformatica "Giulio A. Maccacaro", Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Milano Unità di Post-Genomica, Consorzio Mario Negri Sud, Santa Maria Imbaro, Chieti Università Carlo Cattaneo LIUC, CREMS Centro di Ricerca in Economia e Management in Sanità e nel Sociale, Health Care and Social Management, Castellanza (VA) Università Cattolica del Sacro Cuore di Roma Università dell’Insubria, Varese Università del Piemonte Orientale, Novara Università Milano-Bicocca, Monza Università di Catanzaro Università La Sapienza, Roma Web Medica, Grottaferrata, Roma

INTERNATIONAL COLLABORATIONS

Albert Eistein College of Medicine, Bronx, NY, USA Alzheimer Europe, Bruxelles, Belgium Aston University, Birmingham, UK Beaumont Hospital, Dublin, Ireland Brain Repair Centre, University of Cambridge, Cambridge, UK Brain Research Centre, University of British Columbia, Vancouver, Canada, Cardiovascular and Diabetes Medicine, University of Dundee, UK Cambridge Centre for Brain Repair, University of Cambridge, UK Calouste Gulbenkian Foundation, Lisbona, Portugal Center for Molecular Biology University of Vienna CIMUS Biomedical Research Institute, University of Santiago de Compostela, Spain Chorley & South Ribble General Hospital, Chorley, Cochrane Schizophrenia Group, Università di Nottingham, Nottingham, UK Cochrane Collaboration Depression Anxiety Neurotics Disorders, UK Department of Biochemistry, Boston University, Boston USA Department of Genetics, University of Leicesterm, Leicester,UK Department of Internal Medicine, Hospitales Universitarios Virgen del Rocío, Sevilla, Spain Department of Neurology, University of Lille, France Department of Neuroscience, Karolinska Institute of stocholm , Sweden Department (Neuro) Pathology, Academisch Medisch Centrum ,Amsterdam, The Netherlands Department of Pharmacology and Therapeutics, State University of Maringá, Maringá, PR, Brasil Department of Pediatrics, Neurology Division, David Geffen School of Medicine, UCLA, LA, CA, USA Department of Psychiatry, Geneva University Hospitals, Ginevra, Switzerland Division of Anaesthesia, University of Cambridge, Addenbrooke’s Hospital, Cambridge, UK Division of Medical Genetics, CHUV Lausanne, Switzerland Division of Polymer Chemistry, Department of Chemistry-Ångström, Uppsala University, Uppsala, Sweden Divisione di Geriatria, Ospedali Regionali di Lugano e Mendrisio, Switzerland Epidemiological and Social Psychiatric Research Institute, Erasmus MC, Parnassia Psychiatric Institute EPITARGET Consortium (FP7 EU programme 2013-2018) Hoffmann-La Roche AG, Switzerland HSPH Harvard University, Boston, USA IBCM, University of Lausanne, Lausanne, Switzerland INSERM U 751, Marseille, France Institut d’Investigació Biomèdica de Girona, Girona, Catalunya Institut National de la Santé et de la Recherche Médicale, Paris, France Institute of Biomedical Engineering, University of Oxford, Oxford, UK Institute for Stroke and Dementia Research (ISD) University Hospital Monaco, Germany International Early Psychosis Association King’s College Hospital, London, UK Lancaster University, Lancaster, UK. National Insitute on Aging, NIH, Baltimore, USA National Institutes of HealthsRocky Mountain Laboratories, Montana, USA Neurological Department of the University of Tirana, Albania Neurology, GlaxoSmithKline, New Frontiers Science Park North, Harlow UK Neuroscience Center, Univeristè de Geneve, Switzerland Ninewells Hospital and Medical School, Dundee, Scozia, UK Northern Illinois University, DeKalb, IL, USA Novartis Pharma, Basel, Switzerland NYU, NY, USA Omeros Corporation, Seattle, USA Queen Mary University of London Royal Manchester Children's Hospital, Manchester, UK Royal Preston Hospital, Preston, UK

School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Brasile Sección de Innovación Tecnológica, Hospital Universitario Virgen Macarena y Hospital Universitario Virgen del Rocío, Sevilla, Spain Sergievsky Center, Columbia University, New York, NY, USA Servicio Andaluz de Salud, Sevilla, Spain Service de Neurologie Pédiatrique et des Maladies Métaboliques - INSERM U676, Hôpital Robert Debré - AP-HP – Paris, France Servizio di Geriatria, Ospedale della Beata Vergine, Mendrisio, Switzerland Sheffield Care and Research Centre for Motor Neuron Disorders, University of Sheffield, UK Technological Park of BizkaiaBizkaia, Spain The Scripps Research Institute, Jupiter, Florida, USA Technology Park of Bizkaia, Bizkaia, Spain Toxicology Unit MRC, Leicester, UK Trinity College Dublin e Memory Clinic del St. James’s Hospital di Dublino, Ireland Universidade Nova de Lisboa, International Master in Mental Health Policy and Services, Lisbona, Portugal Universitat de Valencia, Spain Université de la Méditerranée -Hôpital de la Timone Marseille, France University College London, London, UK University of Auckland, New Zealand University Of Barelona, Spain University of Budapest, Hungary Univ of California at Irvine, Irvine, CA, USA University of Cardiff, UK University of Copenhagen, Denmark University Hospital, London, ON, Canada University of Innsbruck, Innsbruck, Austria University of Leicester, UK University of Liverpool, Liverpool, UK University of Maastricht, The Netherlands Università of New South Wales, Australia University of Rijeka Medical School, Rijeka, Croazia University of Rochester, Rochester, NY, United States University of Turku, Turku PET Centre, Turku, Finland University of Utrecht, the Netherlands Université Victor Segalen, Bordeaux, France University of Wroclaw Family Medicine Department, Wroclaw, Poland Unit of Molecular Genetics, CHUV Lausanne, Switzerland Vaccinex company (Rochester,NY,USA) Virtanen Institute for Molecular Sciences, University of Kuopio, Finland Walton Hospital, Liverpool, UK Weizmann Institute, Rehovot, Israel World Association for Psychosocial Rehabilitation World Health Organization, Disability and Rehabilitation Team

EDITORIAL BOARD MEMBERSHIP

Amyotrophic Lateral Sclerosis (Beghi) Annals Pharmacotherapy (Nobili) Biochemical Journal (Chiesa) Clinical Drug Investigation (Beghi) Clinical Neurology and Neurosurgery (Beghi) CNS & Neurological Disorders - Drug Targets (Bendotti) Cochrane Collaboration, Epilepsy (Beghi) Conference Papers in Pharmacology (Nobili)

Dialogo sui Farmaci (Nobili) Drugs in the R&D (Beghi) Early Intervention in Psychiatry (Barbato) Educazione Sanitaria e Promozione della Salute (Barbato) Epidemiologia e Prevenzione (Lucca) Epigenetic of Neurodegenerative Diseases (Forloni) Epilepsia (Beghi) Epilepsy Research (Vezzani) Epilepsy & Treatment (Vezzani) European Child and Adolescent Psychiatry (Barbato) European Journal of Internal Medicine (Nobili) Frontiers in Immunology: Frontiers in Molecular Innate Immunity (De Simoni) Inpharma (Beghi) Intensive Care Medicine experimental (De Simoni senior Zanier associate editor) International Journal of Cell Biology (Chiesa, Guest Editor) International Journal of Mental Health (Barbato) International Journal of Molecular Epidemiology and Genetics (Forloni, senior, Albani associate) International Journal of Neurological Disorders and Interventions (Forloni) ISRN Vascular Medicine (De Simoni) Journal of Alzhiemer’s disease (Albani, Borsello, Forloni) MAP Kinase (Borsello) Neuroepidemiology (Beghi) Neurological Sciences (Beghi) Neuroscience (Vezzani) Open Aging Journal (Forloni) Open Journal of Neuroscience (Forloni) PlosOne (Forloni, Chiesa, Accademic editors) Psychiatric Rehabilitation Journal (Barbato) Quality of Life Research (Barbato) Ricerca & Pratica (Nobili) Sistema Salute (Barbato) The Open Drug Metabolism Journal (Nobili) The Open Pathology Journal (De Simoni) The Scientific World Journal (Nobili) WebmedCentral Pharmacology (Nobili) World Journal of Pharmacology (Nobili) World Journal of Psychiatry(Barbato)

PEER REVIEW ACTIVITIES Acta Neurologica Scandinavica Acta Psychiatrica Scandinava Addiction Biology Age Age and Ageing Alzheimer's & Dementia Alzheimer Disease and Associated Disorders American Journal of Clinical Nutrition American Journal of Hematology American Journal of Human Genetics American Journal of Pathology American Journal of Physiology Amyotrophic Lateral Sclerosis

Annals of Neurology Annals of Pharmacotherapy Archives of Internal Medicine Arthritis Research & Therapy Behavioural Brain Research Behavioural Neuroscience Behavioural Pharmacology Biochimica et Biophysica Acta Biochemical Journal Biochemistry BioMed Central Neurology Biomaterials Biological Psychiatry BMC Geriatrics BMC Neurology BMC Psychiatry BMC Public Health Brain Brain Research Brain Research Bulletin Brain Research Review Canadian Journal of Neurological Sciences Clinical Drug Investigation Clinical Neurology and Neurosurgery Clin Pharm Therapy CNS Drugs Chronobioly International Dementia and Geriatric Cognitive Disorders Drugs Epidemiologia e Psichiatria Sociale Epidemiology and Psychiatric Sciences Epidemiology and Infection Epilepsia Epilepsy & Behavior Epilepsy Research European Journal of Clinical Pharmacology European Journal of Immunology European Journal of Internal Medicine European Journal of Neurology European Journal of Neuroscience European Journal of Pharmacology European Journal of Public Health Experimental Neurology European Neuropsychopharmacology Expert Opinion on Pharmacotherapy European Psychiatry Expert Review in Neurotherapeutics FASEB Journal FEBS letters Fundamental Clinical Psychopharmacology Future Drugs Giornale di Neuropsichiatria dell’Età Evolutiva Glia Global Health Promotion Health and Quality of Life Outcomes Human Molecular Genetics

Intensive Care Medicine International Journal of Mental Health Systems International Journal of Neuropsychopharmacology Journal of Alzhiemer’s disease JAMA Journal of the American Board of Family Practice Journal of Biological Chemistry Journal of Cell. Biology Journal of Cell Physiology Journal of Cerebral Blood Flow and Metabolism Journal of Chemical Neuroanatomy Journal of Clinical Pharmacy and Therapeutics Journal of Clinical Psychiatry Journal of Epidemiology & Community Health Journal of Functional Food Journal of Geriatric Psychiatry and Neurology Journal of Gerontology Journal of Headache and Pain Journal of Histochemistry and Cytochemistry Journal of Immunology Journal of Internal Medicine Journal of Neurochemistry Journal of Neuroimmunology Journal of Neurology, Neurosurgery and Psychiatry Journal of Neuroscience Journal of Pediatric Neurology Journal of Pharmacology and Experimental Therapeutics Journal of Pharmacy and Pharmacology Journal of Psychopharmacology Journal of Psychosomatic Research Journal of Structural Biology Journal of the American Board of Family Practice Journal of Virology Life Sciences Lancet Lancet Neurology Molecular Brain Research Molecular and Cellular Neuroscience Molecular Therapy Nature Clinical Practice Neurology Nature Neuroscience Nature Reviews Neurology Neurobiology of Learning and Memory Neurobiology of Aging Neurobiology of Disease Neuroepidemiology Neurology Neurological Sciences Neuromuscular Disorders Neuropharmacology Neuropsychopharmacology Neuroscience Neuroscience & Neurobehavioral Reviews Neuroscience Letters Neurotherapeutics Neurotoxicity Research

Nordic Journal of Psychiatry N.S. Archives Pharmacology Parkinsonism & Related Disorders Pharmacological Reports Pharmacological Research Pharmacoepidemiology and Drug Safety Pharmacology Biochemistry & Behavior PloS Biology PloS One PloS Pathogens Proc Natl Acad Sci, USA Progress in Neuro-Psychopharmacology & Biological Research Psychiatric Services Psychopharmacology Seizure Schizophrenia Research Social Psychiatry and Psychiatric Epidemiology Stroke Synapse Trends Molecular Medicine The International Journal of Neuropsychopharmacology Vaccine

NATIONAL AND INTERNATIONAL COMMITTEE MEMBERSHIP

Agenzia Europea di Valutazione dei Medicinali (EMEA) Agenzia Italiana per il Farmaco (AIFA) Associazione Italiana di Neuroepidemiologia (Presidente uscente) Associazione Italiana per la Ricerca sull’Invecchiamento Cerebrale (AIRIC, Presidenza) Benchmark’s stewards of the American Epilepsy Society (AES) Board of "Master in Advanced Technologies for the Study of Neurodegenerative Diseases", Milan University Board Assessorato alle Politiche Sociali e Cultura della Salute, Comune di Milano Comitato di coordinamento internazionale del progetto europeo”Quelles professionnalités en santé mentale. Comitato Etico Milano Area B Fondazione IRCCS "Cà Granda" Ospedale Maggiore Policlinico, A.O. Fatebenefratelli ed Oftalmico, A.O. Istituto Ortopedico “G. Pini”, ASL Milano. Perspectives croisées, usagers, élus professionnels”. Commissione di Epidemiologia dell’ILAE (Co-Chair) Commission on European Affairs (CEA, ILAE) Commissione sulla Health Care Policy della Lega Internazionale contro l’Epilessia (ILAE) Comitato Ordinatore del Master in "Tecnologie Avanzate Applicate alle Patologie Neurodegenerative", Università di Milano Comitato scientifico Congresso Mondiale Società per gli Interventi Precoci nelle Psicosi, International Early Psychosis Association Comitato Scientifico AISLA Comitato Scientifico SINDEM Committee for Proprietary Medicinal Products (CPMP) presso L’EMEA Committee for ENCALS Young Investigator Award Consiglio di amministrazione, Fondazione Cecchini Pace, Milano Consiglio Direttivo AIRIC Coordination Group IMI-PharmaCog project Direttivo della Lega Italiana contro l’Epilessia (LICE) Editorial Committee, Guidelines of community based rehabilitation, World Health Organization. Esperto Nazionale, accreditato dall’AIFA (Agenzia Italiana del Farmaco), per l’EMEA (Esperto per il

Etical Committe of the "Area B of Milan": Fondazione IRCCS "Cà Granda" Ospedale Maggiore Policlinico, A.O. Fatebenefratelli ed Oftalmico, A.O. Istituto Ortopedico “G. Pini”, ASL Milano. Medical Research Council (MRC), UK Gruppo di Approfondimento Tecnico per lo sviluppo dell’area ‘Promozione della salute mentale’, Regione Lombardia Gruppo di lavoro sull'epilessia dell'Organizzazione Mondiale della Sanità Gruppo di Studio sull’Epilessia della Società Italiana di Neurologia (SIN) Gruppo di Studio sulla Qualità della Vita della Società Italiana di Neurologia (SIN) Gruppo di Studio sulla Sclerosi Laterale Amiotrofica della Società Italiana di Neurologia (SIN) Medical Research Council Strategic Grant Application, UK Mental Health Working Party, gruppo di lavoro nominato dal Direttorato Generale per la Protezione del Consumatore della Commissione Europea (DG-SANCO), Bruxelles. Gruppo di coordinamento Neuroprion NoE, EU International Committee su “Epilepsy and the Law” International Organizing Committee e coordinator della segreteria al Global Forum for Community Mental International Affair Commission (ILAE) Health, istituito dal Department of Mental Health della World Health Organization. International Subcommittee della American Academy of Neurology International Steering Committee dell’European Network on mental health promotion and mental disorder prevention (EMHPA). International Subcommittee dell’American Academy of Neurology National Institutes of Health of the USA and World Health Organization supported project on The Future of Psychiatric Diagnosis: Refining the Research Agenda. Neurobiology Commission of the International League Against Epilepsy Neuroepidemiology Section of the American Academy of Neurology (Chair uscente) Research Advisory Panel, MND Association, UK Task Force sull’epidemiologia dell’epilessia della ILAE Scientific Advisory Board of Sheffield Institute Foundation for MND Scientific Advisory Board del Thierry Latran Foundation, Francia Scientific Advisory Board of ENCALS (European Network for the Cure of ALS Traslational Research Committee of AES Working Group on Epilepsy della World Health Organization (WHO)

EVENT ORGANIZATION

13a Giornata di studio sulla malattia di Alzheimer: Psicologia dell’invecchiamento: Le funzioni cognitive nei centenari 14 marzo 2015, Ateneo Veneto, Venezia (Lucca) International REPOSI Seminar 2015 Targeting the burden of polypharmacy in the elderly Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico. Milano, 24-25 Settembre 2015 (Nobili) Corso SIMI: Corso introduttivo alla gestione della complessità del paziente anziano ospedalizzato con polipatologia e politerapia. Milano, 1-3 Dicembre 2015 (Nobili)

GRANTS AND CONTRACTS Abbott GmbH & Co. KG ADDF,USA AFM, France Agenzia di Sanità Pubblica del Lazio

AIFA AiRett ALS Society for Canada AriSLA Alzheimer's Association Amgen, Milano AriSLA ASL 2 Piemonte. ASL TO1 Torino Assessorato alla Salute, Comune di Milano Association pour la recherché sur la SLA, France Auris medical Azienda USL 3 Pistoia e Valdinievole Bristol-Myers Squibb Boehringer Ingelheim Centro Studi in Psichiatria ASL TO2, Torino CPADs EU grant CURE Czech Science Foundation EISAI Epilepsy European Research Area Board – ERAB Dana Foundation Dipartimento di Salute Mentale, Azienda Ospedaliera Niguarda Ca’ Granda, Milano Dipartimento di Salute Mentale di Pistoia e Valdinievole Evidentia Medica, Grottaferrata (Roma) Fondazione Cariplo, Milano Fondazione Mariani, Milano Fondazione Italo Monzino, Milano Fondazione Sestini, Italia Fondazione Vialli e Mauro per la Ricerca German-Israeli Foundation for Scientific Research and Development Glaxo-SmithKline, Italy Grünenthal, Germany Hospice "via di Natale Franco Gallini", Aviano (PN) Human Frontiers Scientific Programme IMPHA II, DG-SANCO, Public Health and Consumers' Protection (Directorate General Istituto Comprensivo Statale "G.D. Romagnosi", Carate Brianza (MI) Istituto Regionale Lombardo di Formazione per l’Amministrazione Pubblica – IREF I.R.I.S Istituto San Paolo; Torino Istituto Superiore di Sanità Janssen-Cilag H. Lundbeck A/S, Danimark Hoffmann-La Roche AG, Svizzera Metis, Società Scientifica FIMMG Ministero della Ricerca Scientifica Ministero della Salute NIH-NINDS MND Association, UK Newron Ospedale “Casa Sollievo” di San Giovanni Rotondo Pharming Provincia Autonoma di Trento Progetto Itaca, Milano Regione Lombardia, Assessorato alla Famiglia e Solidarietà Sociale e Assessorato alla Sanità, Milano Rimoldi e Bergamini

Rotary Clubs Gruppo 1, Milano Rotary Clubs Milano Naviglio Grande San Carlo, Milano Scala, Inner Wheel Milano San Carlo Rotta-Pharm, Italy Sanofi-Aventis San Paolo Foundation SELECTA MEDICA, Pavia Servier Laboratories, Parigi Sienabiotech Sinthopharm Thierry Latran Foundation, France Telethon Unione Nazionale Associazioni per la Salute Mentale – UNASAM US-Israel Binational Science Foundation Vertex WebMedica, Grottaferrata (Roma). World Health Organisation

SCIENTIFIC PUBLICATIONS (2015)

Albani D, Mazzuco S, Chierchia A, Fusco F, Boeri L, Martines R, Giorgi ED, Frigato A, Durante E, Caberlotto L, Zanardo A, Siculi M, Gallucci M, Forloni G.The SIRT1 promoter polymorphic site rs12778366 increases IL-6 related human mortality in the prospective study "Treviso Longeva (TRELONG)". Int J Mol Epidemiol Genet. 2015; 6:20-6 Antoniazzi S, Chiarelli MT, Nobili A, Pasina L, Venturini F. The value of software that provides clinically relevant information on drug interactions. Eur J Intern Med. 2015 Nov;26(9):e52-3 Avanzini F, Mafrici A, Riva E, Franzosi MG, Milani Va, Giudici V, Marelli G, Mariani G, Piatti PM, Roncaglioni MC on behalf of GLICINE-SPIDER Collaborative Group A multicenter observational study on the management of hyperglycemia in patients with acute coronary syndrome Project. Nutr Metab Cardiovasc Dis 2015; 25: 916-925. Avanzini F, Marelli G, Saltafossi D, Longhi C, Carbone S, Carlino L, Planca, E Vilei V, Roncaglioni MC, Riva E. Effectivness Safety and feasibility of an evidence-based insulin infusion protocol targeting moderate glycemic control in intensive cardiac care unit. Eur Heart J: Acute Cardiovascular Care 2015; E-pub. Balducci C, Paladini A, Micotti E, Tolomeo D, La Vitola P, Grigoli E, Richardson JC, Forloni G. The Continuing Failure of Bexarotene in Alzheimer's Disease Mice. J Alzheimers Dis. 2015; 46:471-82 Baron R, Ferriero DM, Frisoni GB, Bettegowda C, Gokaslan ZL, Kessler JA, Vezzani A, Waxman SG, Jarius S, Wildemann B, Weller M. Neurology-the next 10 years. Nat Rev Neurol. 2015;11: 658-64. Batelli S, Invernizzi RW, Negro A, Calcagno E, Rodilossi S, Forloni G, Albani D.The Parkinson's disease-related protein DJ-1 protects dopaminergic neurons in vivo and cultured cells from alpha-synuclein and 6-hydroxydopamine toxicity. Neurodegener Dis. 2015; 15:13-23 Bazzoni G, Marengoni A, Tettamanti M, Franchi C, Pasina L, Djade C D, Fortino I, Bortolotti A, Merlino L, Nobili A. The drug prescription network: a system-level view of drug co-prescription in community-dwelling elderly people. Rejuvenation Res 2015; 18: 153-161. Beghi E,. Methodological Demands on Observational Studies of Outcomes After Epilepsy Surgery. In: Long-Term Outcomes of Epilepsy Surgery in Adults and Children. Kristina Malmgren, Sallie Baxendale, J. Helen Cross (Editors); Springer International Publishing Switzerland , 2015; pg. 5-18.

Beghi E, Giussani G, Sander JW. The natural history and prognosis of epilepsy. Epileptic Disord 2015; 17: 243-253. Bellelli G, Nobili A, Annoni G, Morandi A, Djade C D, Meagher D J, Maclullich A M J, Davis D, Mazzone A, Tettamanti M, Mannucci P M, REPOSI Investigators. Under-detection of delirium and impact of neurocognitive deficits on in-hospital mortality among acute geriatric and medical wards. Eur J Intern Med 2015; 26: 696-704. Benedetti MG, Ferrarin M, Cutti G, B E. Conferenza Nazionale di Consenso. Appropriatezza clinica e metodologica dell’analisi strumentale del cammino (“Gait Analysis”) con particolare riferimento alle applicazioni in Medicina Riabilitativa. Giornale Italiano di Medicina Riabilitativa 2015; 28 : 1-9. Benedetti MD, Pugliatti M, D'Alessandro R, Beghi E, Chiò A, Logroscino G, Filippini G, Galeotti F, Massari M, Santuccio C, Raschetti R; ITANG Study Group. A Multicentric Prospective Incidence Study of Guillain-Barré Syndrome in Italy. The ITANG Study. Neuroepidemiology 2015; 45: 90-99.

Benussi L, Rossi G, Glionna M, Tonoli E, Piccoli E, Fostinelli S, Paterlini A, Flocco R, Albani D, Pantieri R, Cereda C, Forloni G, Tagliavini F, Binetti G, Ghidoni R C9ORF72 hexanucleotide repeat number in frontotemporal lobar degeneration: a genotype-phenotype correlation study. J Alzheimers Dis. 2015;45:319 Bergin P, Jayabal J, Walker E, Davis S, Jones P, Dalziel S, Yates K, Thornton V, Bennett P, Wilson K, Roberts L, Litchfield R, Te Ao B, Parmer P, Feigin V, Jost J, Beghi E, Rossetti AO. Use of the EpiNet database for observational study of status epilepticus in Auckland, New Zealand. Epilepsy Behav 2015; 49:164-169. Bouybayoune I, Mantovani S, Del Gallo F, Bertani I, Restelli E, Comerio L, Tapella L, Baracchi F, Fernández-Borges N, Mangieri M, Bisighini C, Beznoussenko GV, Paladini A, Balducci C, Micotti E, Forloni G, Castilla J, Fiordaliso F, Tagliavini F, Imeri L, Chiesa R.Transgenic fatal familial insomnia mice indicate prion infectivity-independent mechanisms of pathogenesis and phenotypic expression of disease.PLoS Pathog. 2015 ;11(4):e1004796. Capovilla G, Beccaria F, Beghi E, Giussani G, Magaudda A, Magaudda L, Beltrami G, Bruttini F, Casasco M, Guerra E, Ieracitano V, Pezzano A, Verzelletti A, Veicsteinas A. EPILESSIA E ATTIVITÀ SPORTIVE. Raccomandazioni congiunte della Lega Italiana contro l’Epilessia (LICE) e della Federazione Medico Sportiva Italiana (FMSI). Medicina dello Sport 2015; 68 (Suppl. 1al N. 1): 5-22. Caproni S, Bianchi E, Cupini LM, Corbelli I, Beghi E, Calabresi P, Sarchielli P; SAMOHA Study Group. Migraine-specific quality of life questionnaire and relapse of medication overuse headache. BMC Neurol 2015; 15: 85. Carli M, Kostoula C, Sacchetti G, Mainolfi P, Anastasia A, Villani C, Invernizzi RW. Tph2 gene deletion enhances amphetamine-induced hypermotility: effect of 5-HT restoration and role of striatal noradrenaline release. J Neurochem 2015, 135:674-685 Chiesa R. (2015). The elusive role of the prion protein and the mechanism of toxicity in prion disease. PLoS Pathogens 11(5):e1004745 Cimini S., Sclip A., Mancini S., Colombo L., Messa M., Cagnotto A., Di Fede G., Tagliavini F., Salmona M., and Borsello T. (2015) Neurobiol Dis. pii: S0969-9961(15)30115-7. Costa DD, Beghi E,. Carignano P, Pagliacci C, Faccioli F, Pupillo E, Messina P, Gorio A, Redaelli T. Tolerability and efficacy of erythropoietin (EPO) treatment in traumatic spinal cord injury: a preliminary randomized comparative trial vs. methylprednisolone (MP). Neurol Sci 2015; 36: 1567-1574.

De Paola M, Sestito SE, Mariani A, Memo C, Fanelli R, Freschi M, Bendotti C, Calabrese V, Peri F. Synthetic and natural small molecule TLR4 antagonists inhibit motoneuron death in cultures from ALS mouse model. Pharmacol Res. 2015 Nov 27;103:180-187. Fabbri C, Crisafulli C, Gurwitz D, Stingl J, Calati R, Albani D, Forloni G, Calabrò M, Martines R, Kasper S, Zohar J, Juven-Wetzler A, Souery D, Montgomery S, Mendlewicz J, Girolamo GD, Serretti A Neuronal cell adhesion genes and antidepressant response in three independent samples. Pharmacogenomics J. 2015;15:538-48 Forloni G, Tettamanti M, Lucca U, Albanese Y, Quaglio E, Chiesa R, Erbetta A, Villani F, Redaelli V, Tagliavini F, Artuso V, Roiter I. Preventive study in subjects at risk of fatal familial insomnia: Innovative approach to rare diseases. Prion 2015; 9: 75-79.

Franchi C, Ardoino I, Rossio R, Nobili A, Biganzoli EM, Marengoni A, Marcucci M, Pasina L, Tettamanti M, Corrao S, Mannucci PM; REPOSI Investigators. Prevalence and Risk Factors Associated with Use of QT-Prolonging Drugs in Hospitalized Older People. Drugs Aging. 2015 Dec 22. [Epub ahead of print] PubMed PMID: 26693921. Franchi C, Baviera M, Sequi M, Cortesi L, Tettamanti M, Roncaglioni M C, Pasina L, Djade C D, Fortino I, Bortolotti A, Merlino L, Mannucci P M, Nobili A. Comparison of health care resource utilization by immigrants versus native elderly people. J Immigr Minor Health 2015, E-pub. Franchi C, Marcucci M, Mannucci PM, Tettamanti M, Pasina L, Fortino I, Bortolotti A, Merlino L, Nobili A. Changes in clinical outcomes for community-dwelling older people exposed to incident chronic polypharmacy: a comparison between 2001 and 2009. Pharmacoepidemiol Drug Saf. 2015 Dec 21. doi: 10.1002/pds.3938. [Epub ahead of print] PubMed PMID: 26687829 Fumagalli S, Perego C, Pischiutta F, Zanier ER, De Simoni MG. The ischemic environment drives microglia and macrophage function. Front Neurol. 2015 Apr 8;6:81 Gervasoni E, Cattaneo D, Messina P, Casati E, Montesano A, Bianchi E, Beghi E. Clinical and stabilometric measures predicting falls in Parkinson disease/parkinsonisms. Acta Neurol Scand 2015; 132: 235-241. Global Burden of Disease Study 2013 Collaborators Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet 2015; 386(9995): 743-800. Global Burden of Disease Study 2013 DALYs and HALE Collaborators. Global, regional, and national disability-adjusted life years (DALYs) for 306 diseases and injuries and healthy life expectancy (HALE) for 188 countries, 1990-2013: quantifying the epidemiological transition. Lancet 2015;386(10009):2145-2191. Giussani G, Canelli V, Bianchi E, Franchi C, Nobili A, Erba G, Beghi E; EPIRES Group. A population-based study of active and drug-resistant epilepsies in Northern Italy. Epilepsy Behav 2016; 55:30-37.

Guaita A, Vaccaro R, Davin A, Colombo M, Vitali SF, Polito L, Abbondanza S, Valle E, Forloni G, Ferretti VV, Villani S. Influence of socio-demographic features and apolipoprotein E epsilon 4 expression on the prevalence of dementia and cognitive impairment in a population of 70-74-year olds: the InveCe.Ab study. Arch Gerontol Geriatr. 2015; 60:334-43

Guekht A, Mizinova M, Ershov A, Guz D, Kaimovsky I, Messina P, Beghi E,. In-hospital costs in patients with seizures and epilepsy after stroke. Epilepsia 2015; 56: 1309-131

Infarinato F, Rahman A, Del Percio C, Lamberty Y, Bordet R, Richardson JC, Forloni G, Drinkenburg W, Lopez S, Aujard F, Babiloni C, Pifferi F; IMI project "PharmaCog" Consortium.

On-Going Frontal Alpha Rhythms Are Dominant in Passive State and Desynchronize in Active State in Adult Gray Mouse Lemurs. PLoS One. 2015 Nov 30;10(11): e0143719.

Iraci N., Stincardini C., Barreca M.L., Biasini E.Decoding the function of the N-terminal tail of the cellular prion protein to inspire novel therapeutic avenues for neurodegenerative diseases Virus Res 2014 ; E-pub Jette N, BeghiE, Hesdorffer D, Moshé SL, Zuberi SM, Medina MT, Bergen D.ICD coding for epilepsy: Past, present, and future-A report by the International League Against Epilepsy Task Force on ICD codes in epilepsy. Epilepsia 2015; 56: 348-355. Khazaal Y, Chatton A, Dieben K, Huguelet P, Boucherie M, Monney G, Lecardeur L, Salamin V, Bretel F, S, Pesenti E, Krychowski R, Costa Prata A, Bartolomei J, Brazo P, Traian A, Charpeaud T, Murys E, Poupart F, Rouvière S, Zullino D, Parabiaghi A, Saoud M, Favrod J Reducing delusional conviction through a cognitive-based group training game: a multicentre randomized controlled trial. Front. Psychiatry 6:66. Lauranzano E, Pozzi S, Pasetto L, Stucchi R, Massignan T, Paolella K, Mombrini M, Nardo G, Lunetta C, Corbo M, Mora G, Bendotti C, Bonetto V. Peptidylprolyl isomerase A governs TARDBP function and assembly in heterogeneous nuclear ribonucleoprotein complexes. Brain. 2015; 138 (Pt 4):974-91 Llovera G, Hofmann K, Roth S, Salas-Pérdomo A, Ferrer-Ferrer M, Perego C, Zanier ER, Mamrak U, Rex A, Party H, Agin V, Fauchon C, Orset C, Haelewyn B, De Simoni MG, Dirnagl U, Grittner U, Planas AM, Plesnila N, Vivien D, Liesz A. Results of a preclinical randomized controlled multicenter trial (pRCT): Anti-CD49d treatment for acute brain ischemia. Sci Transl Med. 2015; 7(299):299ra121. Lucca U, Tettamanti M, Logroscino G, Tiraboschi P, Landi C, Sacco L, Garrì M, Ammesso S, Bertinotti C, Biotti A, Gargantini E, Piedicorcia A, Nobili A, Pasina L, Franchi C, Djade CD, Riva E, Recchia A. Prevalence of dementia in the oldest old: The Monzino 80-plus population based study. Alzheimer's & Dementia 2015; 11: 258-270. Macrì S, Ceci C, Proietti Onori M, Invernizzi RW, Bartolini E, Altabella L, Canese R, Imperi M, Orefici G, Creti R, Margarit I, Magliozzi R Laviola G. Mice repeatedly exposed to group-A b-Haemolytic Streptococcus show perseverative behaviors, impaired sensory motor gating, and immune activation in rostral diencephalon. Sci Rep. 2015, 5:13257 Magini A., Polchi A., Tozzi A., Tancini B., Tantucci M., Urbanelli L., Borsello T., Calabresi P., Emiliani C. Abnormal cortical lysosomal β-hexosaminidase and β-galactosidase activity at post-synaptic sites during Alzheimer's disease progression. (2015) Int J Biochem Cell Biol. 58:62-70. Marino M, Papa S, Crippa V, Nardo G, Peviani M, Cheroni C, Trolese MC, Lauranzano E, Bonetto V, Poletti A, DeBiasi S, Ferraiuolo L, Shaw PJ, Bendotti C. Differences in protein quality control correlate with phenotype variability in 2 mouse models of familial amyotrophic lateral sclerosis. Neurobiol Aging. 2015; 36:492-504 Mandelli S, Riva E, Tettamanti M, Detoma P, Giacomin A, Lucca U. Mortality prediction in the oldest old with five different equations to estimate glomerular filtration rate: The Health and Anemia population-based Study. PLoS ONE 28 Aug, 2015. DOI:10.1371/journal.pone.0136039

Mannucci PM, Nobili A. Appropriateness of antithrombotic prophylaxis in the oldest old with non-valvular atrial fibrillation: ARAPACIS and REPOSI. Eur J Intern Med. 2015 Nov;26(9):e47-8. doi: 10.1016/j.ejim.2015.08.013. Epub 2015 Sep 5. PubMed PMID: 26350113.

Marelli G, Avanzini F, Iacuitti G, Planca E, Frigerio I, Busi G, Carlino L, Cortesi L, Roncaglioni MC, Riva E. Effectiveness of a nurse-managed protocol to prevent hypoglycemia in hospitalized patients with diabetes. J Diabetes Res 2015, E-pub. Marengoni A, Nobili A, Corli O, Djade C D, Bertoni D, Tettamanti M, Pasina L, Corrao S, Salerno F, Marcucci M, Mannucci P M, REPOSI Investigators. The stigma of low opioid prescription in the hospitalized multimorbid elderly in Italy. Intern Emerg Med 2015; 10: 305-313. Marengoni A, Nobili A, Onder G. Best Practices for Drug Prescribing in Older Adults: A Call for Action. Drugs Aging. 2015 Nov;32(11):887-90. doi: 10.1007/s40266-015-0324-3. PubMed PMID: 26586379 Micotti E, Paladini A, Balducci C, Tolomeo D, Frasca A, Marizzoni M, Filibian M, Caroli A, Valbusa G, Dix S, O'Neill M, Ozmen L, Czech C, Richardson JC, Frisoni GB, Forloni G.Striatum and entorhinal cortex atrophy in AD mouse models: MRI comprehensive analysis. Neurobiol Aging. 2015; 36:776-88. Mishto M, Raza ML, de Biase D, Ravizza T, Vasuri F, Martucci M, Keller C, Bellavista E, Buchholz TJ, Kloetzel PM, Pession A, Vezzani A, Heinemann U. The immunoproteasome β5i subunit is a key contributor to ictogenesis in a rat model of chronic epilepsy. Brain Behav Immun. 2015;49:188-96. Nobile-Orazio E, Cocito D, Jann S, Uncini A, Messina P, Antonini G, Fazio R, Gallia F, Schenone A, Francia A, Pareyson D, Santoro L, Tamburin S, Cavaletti G, Giannini F, Sabatelli M, BeghiE,; for the IMC Trial Group. Frequency and time to relapse after discontinuing 6-month therapy with IVIg or pulsed methylprednisolone in CIDP. J Neurol Neurosurg Psychiatry 2015; 86: 729-734. Papadimou E, Morigi M, Iatropoulos P, Xinaris C, Tomasoni S, Benedetti V, Longaretti L, Rota C, Todeschini M, Rizzo P, Introna M, De Simoni MG, Remuzzi G, Goligorsky M S, Benigni A. Direct reprogramming of human bone marrow stromal cells into functional renal cells using cell-free extracts. Stem Cell Reports 2015; 4: 685-698 Parabiaghi A, Tettamanti M, D'Avanzo B, Barbato A, GISAS Study Group. Metabolic syndrome and drug discontinuation in schizophrenia: a randomized trial comparing aripiprazole, olanzapine and haloperidol. Acta Psychiatr Scand 2015; E-pub. Parabiaghi A, Barbato A, Risso P, Fortino I, Bortolotti A, Merlino L, D'Avanzo B. Lithium use from 2000 to 2010 in Italy: A population-based study. Pharmacopsychiatry 2015; E-pub

Pasina L, Marengoni A, Ghibelli S, Suardi F, Djade CD, Nobili A, Franchi C, Guerrini G. A Multicomponent Intervention to Optimize Psychotropic Drug Prescription in Elderly Nursing Home Residents: An Italian Multicenter, Prospective, Pilot Study. Drugs Aging. 2015 Dec 21. [Epub ahead of print] PubMed PMID: 26689398.

Pasina L, Zanotta D, Puricelli S, Djignefa DC, Bonoldi G. Proton pump inhibitors and risk of hypomagnesemia. Eur J Intern Med. 2015 Sep;26(7):e25-6. doi: 10.1016/j.ejim.2015.06.019. Epub 2015 Jul 9. PubMed PMID: 26165831.

Pasina L, Urru SAM, Giua C, Minghetti P. Role of community pharmacies for the detection of potentially inappropriate xanthine oxidase inhibitor prescriptions. Drugs - Real World Outcomes 2015; 2: 81-86 Petitbarat M, Durigutto P, Macor P, Bulla R, Palmioli A, Bernardi A, De SimoniMG, Ledee N, Chaouat G, Tedesco F. Critical Role and Therapeutic Control of theLectin Pathway of Complement Activation in an Abortion-Prone Mouse Mating. J Immunol. 2015;195: 5602-7. Piscopo P, Tosto G, Belli C, Talarico G, Galimberti D, Gasparini M, Canevelli M, Poleggi A, Crestini A, Albani D, Forloni G, Lucca U, Quadri P, Tettamanti M, Fenoglio C, Scarpini E, Bruno G, Vanacore N, Confaloni A. SORL1 Gene is Associated with the Conversion from Mild Cognitive Impairment to Alzheimer’s Disease. J Alzheimers Dis 2015; E-pub.

Pupillo E, Bianchi E, Messina P, Chiveri L, Lunetta C, Corbo M, Filosto M, Lorusso L, Marin B, Mandrioli J, Riva N, Sasanelli F, Tremolizzo L, Beghi E and the EURALS Consortium. Extrapyramidal and cognitive signs in amyotrophic lateral sclerosis. Amyotroph Lateral Scler Frontotemporal Degener 2015; 16: 324-330 Relja B., Weber R., Maraslioglu M., Wagner N., Borsello T., Jobin C., Marzi I and Lehnert M.Differential Relevance of NF-κB and JNK in the Pathophysiology of Hemorrhage/Resususcitation-Induced Liver Injury after Chronic Ethanol Feeding. (2015) Plos One. 10(9):e0137875.

Riva S, Nobili A, Djade CD, Mancuso ME, Santagostino E, Pravettoni G. Cognitive and psychological profiles in treatment compliance: a study in an elderly population with hemophilia. Clin Interv Aging. 2015 Jul 9;10:1141-6. doi: 10.2147/CIA.S84749. eCollection 2015. PubMed PMID: 26185433; PubMed Central PMCID: PMC4501679.

Riva S, Nobili A, Pravettoni G. Light and shade behind the opt-out system for the HIV screening. Lancet HIV. 2015 Aug;2(8):e317. doi: 10.1016/S2352-3018(15)00135-6. Epub 2015 Jul 29. PubMed PMID: 26423371. Roseti C, van Vliet EA, Cifelli P, Ruffolo G, Baayen JC, Di Castro MA, Bertollini C, Limatola C, Aronica E, Vezzani A, Palma E. GABAA currents are decreased by IL-1β in epileptogenic tissue of patients with temporal lobe epilepsy: implications for ictogenesis. Neurobiol Dis. 2015;82:311-20. Rossio R, Franchi C, Ardoino I, Djade CD, Tettamanti M, Pasina L, Salerno F, Marengoni A, Corrao S, Marcucci M, Peyvandi F, Biganzoli E M, Nobili A, Mannucci P M, REPOSI Investigators. Adherence to antibiotic treatment guidelines and outcomes in the hospitalized elderly with different types of pneumonia. Eur J Intern Med 2015; 26: 330-337. Santalucia P, Baviera M, Cortesi L, Tettamanti M, Marzona I, Nobili A, Riva E, Fortino I, Bortolotti A, Merlino L, Roncaglioni M C. Epidemiologic trends in hospitalized ischemic stroke from 2002 to 2010: results from a large Italian population based-study. J Stroke Cerebrovasc Dis 2015; 24: 1917-1923. Santalucia P, Franchi C, Djade C D, Tettamanti M, Pasina L, Corrao S, Salerno F, Marengoni A, Marcucci M, Nobili A, Mannucci PM, REPOSI Investigators. Gender difference in drug use in hospitalized elderly patients. Eur J Intern Med 2015; 26: 483-490. Sarchielli P, Corbelli I, Messina P, Cupini LM, Bernardi G, Bono G, Di Piero V, Petolicchio B, Livrea P, Prudenzano MP, Pini LA, Sandrini G, Allena M, Tedeschi G, Russo A, Caproni S, Beghi E, Calabresi P; SAMOHA Study Group. Psychopathological comorbidities in medication-overuse headache: a multicentre clinical study. Eur J Neurol 2016; 23: 85-91. Staszewsky L, Cortesi L, Baviera M, Tettamanti M, Marzona I, Nobili A, Fortino I, Bortolotti A, Merlino L, Disertori M, Latini R, Roncaglioni MC. Diabetes mellitus as risk factor for atrial fibrillation hospitalization: incidence and outcomes over nine years in a region of Northern Italy. Diabetes Res Clin Pract 2015; 109: 476-484. Stocchetti N, Magnoni S, Zanier ER. My paper 20 years later: cerebral venous oxygen saturation studied with bilateral samples in the internal jugular veins. Intensive Care Med. 2015; 41:412-7. Stocchetti N, Taccone FS, Citerio G, Pepe PE, Le Roux PD, Oddo M, Polderman KH, Stevens RD, Barsan W, Maas AI, Meyfroidt G, Bell MJ, Silbergleit R, Vespa PM, Faden AI, Helbok R, Tisherman S, Zanier ER, Valenzuela T, Wendon J, Menon DK, Vincent JL. Neuroprotection in acute brain injury: an up-to-date review. Crit Care. 2015; 19:186. Sukhanov I, Caffino L, Efimova EV, Espinoza S, Sotnikova TD, Cervo L, Fumagalli F, Gainetdinov RR. Increased context-dependent conditioning to amphetamine in mice lacking TAAR1. Pharmacol Res. 2015 Dec 2;103:206-214. doi: 10.1016/j.phrs.2015.11.002. [Epub ahead of print].

Tortarolo M, Vallarola A, Lidonnici D, Battaglia E, Gensano F, Spaltro G, Fiordaliso F, Corbelli A, Garetto S, Martini E, Pasetto L, Kallikourdis M, Bonetto V, Bendotti C.Lack of TNF-alpha receptor type 2 protects motor neurons in a cellular model of amyotrophic lateral sclerosis and in mutant SOD1 mice but does not affect disease progression. J Neurochem. 2015 Oct;135(1):109-24. Tozzi A., Sclip A, Tantucci A., De Lure A., Griglieri V., Costa C., Di Filippo M., Borsello T, Calabresi P. Region- and age-dependent reductions of hippocampal long-term potentiation and NMDA to AMPA ratio in a genetic model of Alzheimer's diseaseNeurobiol Aging. 2015; 36:123-33

Urru SA, Pasina L, Minghetti P, Giua C. Role of community pharmacists in the detection of potentially inappropriate benzodiazepines prescriptions for insomnia. Int J Clin Pharm. 2015; 37: 1004-8.

Valbuena GN, Rizzardini M, Cimini S, Siskos AP, Bendotti C, Cantoni L, Keun HC.Metabolomic Analysis Reveals Increased Aerobic Glycolysis and Amino Acid Deficit in a Cellular Model of Amyotrophic Lateral Sclerosis. Mol Neurobiol. 2015 May 12. [Epub ahead of print] Vallarino M, Henry C, Etain B, Gehue L G, Macneil C, Scott E M, Barbato A, Conus P, Hlastala S A, Fristad M, Miklowitz D J, Scott J An evidence map of psychosocial interventions for the earliest stages of bipolar disorder. Lancet Psychiatry 2015; 2: 548-56 Vercelli A., Biggi S., Sclip A., Repetto I.E., Cimini S., Falleroni F., Tomasi S., Monti R., Tonna N., Morelli F., Grande V., Stravalaci M., Biasini E., Marin O., Bianco F., di Marino D. and Borsello T. The cell-permeable Aβ1-6A2VTAT(D) peptide reverts synaptopathy induced by Aβ1-42wt. Exploring the role of MKK7 in excitotoxicity and cerebral ischemia: a novel pharmacological strategy against brain injury (2015) Cell Death Dis.;6:e1854 Violatto MB, Santangelo C, Capelli C, Frapolli R, Ferrari R, Sitia L, Tortarolo M, Talamini L, Previdi S, Moscatelli D, Salmona M, Introna M, Bendotti C, Bigini P. Longitudinal tracking of triple labeled umbilical cord derived mesenchymal stromal cells in a mouse model of Amyotrophic Lateral Sclerosis. Stem Cell Res. 2015; 15:243-53. Walker A, Russmann V, Deeg CA, von Toerne C, Kleinwort KJ, Szober C, Rettenbeck ML, von Rüden EL, Goc J, Ongerth T, Boes K, Salvamoser JD, Vezzani A, Hauck SM, Potschka H. Proteomic profiling of epileptogenesis in a rat model: Focus on inflammation. Brain Behav Immun. 2015 Dec 11. pii: S0889-1591(15)30069-6. [Epub ahead of print] Weissberg I, Wood L, Kamintsky L, Vazquez O, Milikovsky DZ, Alexander A, Oppenheim H, Ardizzone C, Becker A, Frigerio F, Vezzani A, Buckwalter MS, Huguenard JR, Friedman A, Kaufer D. Albumin induces excitatory synaptogenesis through astrocytic TGF-β/ALK5 signaling in a model of acquired epilepsy following blood-brain barrier dysfunction. Neurobiol Dis. 2015;78:115-25. Zacchetti L, Magnoni S, Di Corte F, Zanier ER, Stocchetti N. Accuracy of intracranial pressure monitoring: systematic review and meta-analysis. Crit Care. 2015;19:420 Zanier ER, Fumagalli S, Perego C, Pischiutta F, De Simoni MG. Shape descriptors of the "never resting" microglia in three different acute brain injury models in mice. Intensive Care Med Exp. 2015; 3: 39. Zoerle T, Lombardo A, Colombo A, Longhi L, Zanier ER, Rampini P, Stocchetti N. Intracranial pressure after subarachnoid hemorrhage. Crit Care Med. 2015; 43: 168-76.

LAY PRESS SELECTION (2015) Ghislandi F, Beghi E,, A. Curto, L. Garattini. Revisione critica delle valutazioni economiche europee sulle terapie consolidate di prima linea per la Sclerosi Multipla. Quaderni di Farmacoeconomia 28 ottobre 2015.

Benedetti MG, Ferrarin M, Cutti G, B E. Conferenza Nazionale di Consenso. Appropriatezza clinica e metodologica dell’analisi strumentale del cammino (“Gait Analysis”) con particolare riferimento alle applicazioni in Medicina Riabilitativa. Giornale Italiano di Medicina Riabilitativa 2015; 28 : 1-9.

RESEARCH ACTIVITIES

Laboratory of Biology of Neurodegenerative Disorders Alzheimer's disease: genetic studies and clinical investigations In collaboration with different neurological centers and the laboratory of Geriatric Neuropsychiatry it has been created a bank of blood samples for DNA of patients with Alzheimer’s disease (AD), in familial (FAD) or sporadic form (SAD), and patients with vascular dementia (VD). In all subjects the diagnosis of dementia is performed according to the international guidelines. Since 2005 we started also the collection of blood samples from subjects with front-temporal dementia. The genetic studies are aimed to the identification of causal factors in FAD and risk factors in SAD. Mutations on genes encoding proteins involved in the physiopathology of AD were investigated. The pathogenic role of these mutations is under investigation using fibroblasts obtained from skin biopsy. Furthermore, we continued the screening of FAD samples for the genes encoding for presenilin 1 and 2 (PS-1 and PS-2) and APP, missense mutations in these three genes were associated with AD. Alzheimer's disease: preclinical studies The formation of amyloid (A) deposits in brain parenchyma and on the wall of cerebral blood vessels is an early event in AD and there are now numerous genetic, biochemical and neuropathological studies pointing to a causal role of A in the pathogenesis of AD. Thus, prevention the formation of A aggregates or their elimination once formed is a potential therapeutic approach to the disease. This aim is strongly persecuted with different strategies including the regulation of enzymes responsible of the synthesis and degradation of A and the enzymes influencing the metabolism of amyloid precursor protein (APP). In the lab, we developed the idea to interfere directly with the A deposits formation using anti-amyloidogenic drugs. The experimental studies have shown the potential therapeutic activity of these drugs in AD, and now they will be tested in a clinical setting. Alzheimer’s disease: Translational studies In the frame of the European Consortium IMI-PharmaCog have been set up several protocols for the MRI analysis in various transgenic mice models of Alzheimer’s disease (AD). The PharmaCog project focused on the optimization of the translational studies to facilitate the therapeutic approaches considering in experimental models and in the clinical studies the same parameters, behaviorally, biochemically and of imaging. In this contest it will be analyzed longitudinally in single, carrying human amyloid precursor protein mutated (APP) associated to AD, double carrying APP and mutated PS1 transgene, and triple transgenic mice carrying APP, PS2 and mutated tau transgene. We performed the MRI analysis in the same animals at 4, 8, 12, 18 and 24 months, the analysis has been structural, functional and spettroscopical. The strumental parameters (ROI, T2, DTI) have been harmonized with the partners developing similar approaches in humans. The analyis in PDAPP mice has been interrupted due to inconsistency of the phenotype. The main result was the progressive reduction with aging of striatal volume and the enterhinal cortex thickness. Also the hippocampal volume was smaller in both double and transgenic mice but in TASTPM (double) the shrinkage of this area was evident from the first months of life while in triple transgenic mice was progressive with age. The reduction of striatal volume and enterhinal cortex thickness has translational meaning since it was found also in the familial form of AD. During 2015 the structural analysis have been associated with fuctional examinations. Nanoparticles in experimental models of Alzheimer’s disease

One of the main problems that need to be addressed in the therapeutic approaches to central nervous system disorders is the passage of blood brain barrier (BBB) of the drugs and substances potentially active. In the last few years has been stressed the possibility that nanoparticles might represent a good vehicle to translate the drugs within the brain. As part of an European Consortium coordinates by Department of Biochemistry of the Bicocca University in Milan we had the possibility to test various types of nanoparticles in experimental model of Alzheimer’s disease. In collaboration with our Department of Biochemistry and the Bicocca University, liposome nanoparticles were functionalized with an ApoE peptidergic fragment and phosphatidic acid that in vitro was able to exert anti-amyloidogenic activity. Transgenic mice expressing human mutated amyloid precursor protein (APP) alone or in combination with PS1 (APP/PS1) were treated intravenously with functionalized liposomes every other day for three weeks. The treatment reduced the sizes of cerebral amyloid plaques and the content of amyloid oligomers, these effects were associated with recovery of cognitive performance determined with approprate tests The role of oligomers in the Alzheimer pathogenesis Recent data have shown the essential role plays by oligomers, small and soluble aggregates of Ain the Alzheimer pathogenesis and in particular in the cognitive decline associated to the disease. In collaboration with the Department of Biochemistry we developed some in vivo models to analyze the neuronal dysfunction induced by Abut not in monomeric or fibrillar species. The intracerebral application of these different forms confirmed that Aoligomers induced behavioral impairment while monomeric or fibrillar forms of Adid not affect the cognitive behavior. More recently it has been investigated the biological mechanisms responsible of the cognitive decline and the role inflammation in this deficit Sirtuins and neurodegeneration The sirtuins are a family of conserved proteins with de-acetylation activity. In human the sirtuins are coded by 7 different genes and are localized in the citosol, within the nuclei and in the cellular mitochondria. SIRT-1, the better known sirtuin, is involved in the aging physiology and energetic metabolism, its activation induced beneficial effects in Alzheimer and Parkinson experimental models. We studied sirtuins from different points of view, genetic, cellular and behaviorally. The genetic studies are devoted to identify alterations associated to AD in Italian populations. During the screening of all sirtuin genes, we found several single nucleic polymorphisms that now are investigated in larger population (560 AD subjects). The cellular studies are focused on the role of SIRT-1 and SIRT-2 in the cell death mechanisms and oxidative stress in cellular models of AD. Since sirtuins have been involved in the energetic metabolism, and mental as well as physical exercise exert protective effect in AD, we are evaluating in AD animal models if sirtuins are able to mediate the beneficial effects of physical exercise and environmental stimulation. Genetics of aging In collaboration with Geriatric Neuropsychiatry Lab for the Monzino 80-plus study and with dr. Maurizio Gallucci from the ARGel Association in Treviso for Trelong study we collected a large number of blood samples from subjects over seventy. In these samples we are performing a genetic analysis to identify genetic profiles associate to the longevity and /or to the aging-associated pathologies with specific attention to the dementias. The aim is to cross the genotype/phenotype profile with pathologies and environmental aspects including style of life, diet and economical conditions to identify risks and protective factors. Initially the subjects were genotypized for ApoE, whom allele E4 is a well-known risk factor for Alzheimer’s disease and several other disorders and sirt-1 a gene codified for protein member of a enzymatic family of sirtuins associated to the longevity in several experimental models. The results are interesting but before drawing any conclusion we need to consider the numerous other parameters collected in our database. Parkinson’s Disease: genetic studies Parkinson’s disease (PD) is the second more diffuse neurodegenerative disorder with an unknown pathogenesis, however for PD several therapies are available and, although at the symptomatic level, their efficacies is well-established. In the etiological studies on PD the genetic component has been traditionally considered with scarce interest whereas the environmental causes were carefully evaluated. This orientation was based on the evidence that the exposure to several toxins can mimic the PD pathology. However the genetic studies in the last few years have completely changed the perspective with the identification of

mutations on two genes, encoding for alpha-synuclein and parkin, associated to the juvenile forms of the disease. A mutation on alpha synuclein gene is an event extremely rare, only three mutations identified until now, the parkin mutations are numerous ether in puntiform or in deletion form. The mutations on alpha-synuclein gene are dominant while the parkin mutations are associated with PD in recessive form. We collected, in collaboration with several neurological centers, blood samples from PD subjects and the screening of the samples involved genes like alpha-synuclein, parkin, DJ-1 and other factors potentially involved in PD. Parkinson’s disease: in vitro studies and in vivo studies The identification of the mutations associated to Parkinson’s disease (PD) gave a substantial contribute to understand the disease and allowed the development of cellular models to investigate the pathogenesis of the disease. In the past we showed the potential neurotoxic activity of alpha-sinuclein using the synthetic peptide homologous to the fibrillogenic fragment 61-95 (NAC) of the protein. Successively with help of dr. Negro at the Department of Biochemistry at the University of Padova we prepared cDNA vectors including the sequence of wild type and mutated alpha-synuclein Their transfection to the PC12 cells induced in specific conditions a cellular damage. More recently in collaboration with the University of Insubria we obtained the synthesis of synuclein wild type and mutated, together with the in vitro experiments where the primary cells were exposed to the synuclein, we have developed a in vivo model similar to that setting up with amyloid oligomer. The small aggregates of -synuclein are injected intraventricularly and the effect of cognitive decline has been evaluated , with different pharmacological interactions are investigated the similarities and the differences from the application of amyloid Spinal injury and regeneration The mission of Spinal Acute Trauma and Regeneration Unit is to develop therapies to contrast the degenerative events associated to acute spinal injury using nanomaterials to a controlled release of drugs and cells. recently in collaboration with the Department of Chemistry, Material and Chemical Engineering "Giulio Natta" at the Polytechnic University of Milan, have been tested polymeric nanoparticles poly-ε-caprolactone and polyethylene glycol to vehicular drugs interfering with the secondary damage after spinal trauma. These nanoparticles are particularly interesting because they apparently interact exclusively with macrophages and microglia cells. In the lab the toxicological aspects have been verified in vitro before to load them with minocycline a tetracycline with anti-inflammatory effect. After in vitro investigations to optimize the proportion minocycline/nanoparticles, the nanoparticles have been implanted in a murine model of spinal injury. The microglial selectivity has been confirmed in vivo and apparently also the functional aspects have been improved with the treatment.

Laboratory of Cell Death and Neuroprotection AD and APP gene mutations APP gene was the first to have been found mutated in an inherited form of AD. In 2009, the group of Prof. Tagliavini has identified a novel mutation in the APP gene in an Italian family that causes disease only in the case of homozygosity. This mutation consists of the substitution of an alanine with a valine at position 673 of the APP (A673V), corresponding to position 2 of the peptide A. In vitro studies showed that the mutation A673V (A2V) moves the processing of APP towards the amyloidogenic pathway by increasing production of A. Also, biochemical analysis of synthetic peptides of -amyloid 1-40 with A2V mutation showed an aggregation kinetics faster than WT. On the contrary the equimolar co-incubation of two species leads to a block of oligomerization, mimicking the heterozygote state. These observations have important implications for the development of a potential new treatment for the familial and sporadic forms of AD, based on modified A peptides. In this regard, short cell-permeable synthetic peptides were generated and they were able to mimic the anti-aggregating effect of the A with the mutation A2V on the WT A. These peptides were made by first six amino acids of mutated Aβ, necessary to maintain the ability to prevent the formation of oligomers and amyloid fibrils in vitro. The aminoacidic residues were conjugated with the TAT sequence of the HIV virus to allow the delivery through cell membranes. We tested the peptide D-TAT 1-6 A2V in vitro and in vivo. In vitro experiments showed that the peptide alone is not toxic and when it is administered in combination with the A WT it is able to prevent alterations in PSD and the decrease in the number of dendritic spines. We obtained the same

results also in a preliminary study in vivo in a mouse model of AD. These results suggest that D-TAT 1-6 A2V peptide could represent a promising strategy to block the progression of AD. JNK’s role in Rett syndrome Rett syndrome (RTT) is a progressive neurodevelopmental disorder with an incidence of 1 in 10000. RTT is caused by heterozygous mutations in the X-linked MECP2 encoding methyl-CpG-bindng protein-2, a transcription factor. MeCP2 regulates activity-dependent synaptic maturation and maintenance. Studies in RTT mice established MeCP2 as a critical mediator of synaptic scaling up and raise the possibility that some of the neurological defects of Rett arise from a disruption of homeostatic plasticity. Rett syndrome appears as a synaptopathy. Analysis of neuron morphology on animal models of RTT showed a significant region-specific reduction in number and length of dendrites. The main topic of this project is the molecular characterization of early signalling pathways that underlies synaptic dysfunction in Rett Syndrome. At this aim we studied the biochemical changes of the PSD region using two different Mecp2 mouse models: 1) Mecp +/-, heterozygous mutation founded in RTT females; 2) Mecp2 Knockout, where the gene is completely knockout in order to study its role in synaptopathy. We will focus our studies on JNK kinase, which has a key role in excitatory synapses dysfunction. Infact many computational biology studies pointed out that JNK can directly interact with MeCP2. We are confirming this interaction through the immuno-precipitation technique, in mouse brain areas involved in sinaptopathy. Furthermore, in order to deeply comprehend JNK’s involvement in pathological events leading to neuronal damage, we will validate cell-permeable peptide D-JNKI1 for the treatment of this pathology and analyze the signalling pathways that underlie synaptic dysfunction in Rett Syndrome. JNK’s role in pre-synaptic vesicles mobilization and glutamate release from axonal ending. Even if a growing number of papers showed JNK kinase’s role in many neurodegenerative processes, such as early AD phases associated dendritic spines loss, its function in the pre-synaptic compartment is still unknown. At this purpose we initially confirmed JNK presence in the axonal ending and then studied its ability to modulate neurotransmitter release. It’s commonly known in fact, that many other kinases are able to phosphorylate pre-synaptic machinery proteins and induce an increase in vesicles fusion at the active zone. We proved that glutamatergic NMDA autoreceptors stimulation provokes a significant increase in pre-synaptic JNK activation and that glutamate release after stimulation is slightly reduced by JNK specific inhibitor (D-JNKI) administration. After an aminoacidic-sequence screening we discovered that SNARE proteins, constitutive elements of synaptic vesicles docking and fusion complex, contain possible JNK binding domains (JBD). Among these we proved that syntaxin-2 and SNAP25, internal membrane t-SNAREs, preferentially interact with JNK in the neurotransmitter release process. In addiction to that we proved that even phosphorylated, and therefore activated, JNK form it’s able to interact with the two targets, confirming the hypothesis that JNK activation after NMDA stimulation leads to an interaction with SNARE proteins. We then focused our attention on JNK isoforms (1,2 and 3) and found that JNK2 and JNK3 preferentially interacts with SNARE proteins. In order to elucidate how JNK is connected to the upstream NMDA receptor we studied calcium ionic currents on functional synaptosomes and found that JNK inhibition significantly reduced calcium efflux after NMDA stimulation. This is probably due to a reduction in NMDAR or voltage dependent calcium channels (VDCC) conductance, as has been proved for many other presynaptic kinases. We will then proceed evaluating JNK’s ability to phosphorylate syntaxin 2 and SNAP25, displacing SNARE complex assembly, and to interact with NMDAR and VDCC. Interaction/phosphorylation domain individuation will open the road to the design of new molecules able to displace this interaction and modulate glutamate release. Excessive glutamate release indeed, is associated to many neurodegenerative processes such as ischemia, epilepsy, AD and psychiatric disorders; for this reason the development of new compounds, aimed at blocking this phenomenon, is a promising research field, full of interesting therapeutic potentialities. Neuroprotective effect of new cell-permeable MKK7 inhibitor peptides in cerebral ischemia models

JNK MAP kinase activation is involved in the excitotoxic and inflammatory events related to early phases of cerebral ischemia. Previous works from our laboratory proved that only MKK7, one of the two JNK’s upstream activators (MKK7 and MKK4), is activated after an ischemic event. We therefore postulated that MKK7 is the only responsible of JNK’s pathological activation, while MKK4 has a homeostatic maintenance function. We hence chose JNK-MKK7 interaction as target for the development of a new therapeutic compound, in order to avoid interference with JNK’s physiological activation and risk of side effects. Modelling design preliminary studies brought us to synthesize a selective MKK7 inhibitor, based on Cell-Permeable Peptides technology, which allows therapeutic peptide to cross cell membranes thanks to its linkage to TAT peptide. Inhibitor molecule has been modelled on MKK7 binding domain site on Gadd45β, a member of NFkB pathway, which specifically blocks MKK7 catalytic activity without interfering with MKK4’s activation. We synthesized two peptides: GADD45β(69-86) which contains only the binding region, and GADD45β(60-86) which also contains a region essential for MKK7 inactivation. We firstly proved with in vitro experiments that the two peptides have no toxic effects on neurons and that their administration has a significant neuroprotective effect in two in vitro models of ischemia (NMDA treatment and Oxygen-Glucose Deprivation). Neuroprotective effect is specifically due to MKK7 inhibition, while MKK4 activation shows no significant reduction. Our promising in vitro results have then been reproduced in two in vivo models of cerebral ischemia and we proved that peptides administration, at different timepoints before and after lesion, induces a slightly reduction of ischemic volume. Evaluating in vivo signalling we confirmed that peptides administration significantly reduce MKK7 activation, without interfering with MKK4. We particularly observed that, 3 hours as well as 6 hours after ischemia, inhibitory effect causes a reduction in JNK’s activation, responsible for the neuroprotective effect of the peptides. Alzheimer’s disease and eye neurodegeneration Alzheimer’s disease leads to eye neurodegeneration at early stages: optical coherence tomography (OCT), a non-invasive test, shows in AD patients a thinning of the retinal nerve fiber layer (RNFL), formed directly from the optic nerve and therefore particularly sensitive to neurodegeneration. This evidence was also observed in CRND8 mice model used in our laboratory for AD: preliminary data show the presence in eye total homogenate of toxic proteins such as APP, P-APP and P-TAU and APP processing towards amyloidogenic pathway with A1-42 accumulation. We therefore observed that chronic treatment with D-JNKI peptide, a specific JNK inhibitor, leads to a reduction in p-APP and p-tau in the optic nerve. This data confirm the strong involvement of JNK kinase in AD pathogenesis, already proved in other brain areas. OCT is a powerful fast non-invasive diagnostic tool for patients: it allows AD early diagnosis, because RNFL thinning is already present in early stages of the disease. Our interest is to investigate, through biochemical and morphological techniques, which eye area is affected by the presence of the toxic proteins that determine the disease, and the efficacy of D-JNKI treatment, in order to discover the molecular mechanisms of the eye neurodegeneration and to inhibit ocular pathological processes. Molecular mechanism in synaptopathy The dysfunction of excitatory synapses is the first toxic event characterizing several neurodegenerative diseases, including Alzheimer's disease (AD). In AD patients there is a decrease in the number of dendritic spines that correlates with cognitive deficits. Soluble forms of oligomeric Aß are responsible of synaptopathy, and interfere with glutamatergic transmission by reducing the levels of glutamate receptors in the postsynaptic compartment, favoring the collapse of dendritic spines. We generated an in vitro model to study the synaptic degeneration and we identified potential protective molecules. Prevention of synaptopathy represents a new strategy for the development of effective therapies for AD treatment, currently incurable. In this model we studied signal transduction pathways activated by oligomeric Aß toxic stimulus and we demonstrated that exposure to oligomers strongly activates JNK kinase at the synaptic level. D-JNKI1, JNK kinase specific inhibitor, protected neurons from synaptic dysfunction, preventing dendritic spines loss and reduction of postsynaptic receptors induced by A� toxicity. Therefore D-JNKI1 is a promising molecule for Alzheimer's disease treatment. We therefore explored the mechanisms by which JNK exerts its function at the synaptic level and we demonstrated that it acts on two postsynaptic targets: caspase-3 and PSD95. JNK promotes the activation of caspase-3 in the postsynaptic compartment, which regulates the internalization of AMPA receptors from the membrane. JNK also interacts with PSD95 at postsynaptic level and phosphorylates it at Ser320 or Thr321. PSD95 phosphorylation results in the removal of the protein from the synaptic compartment. PSD95 is a scaffold protein that maintains the organization of dendritic spine,

recruiting glutamate receptors; for this reason its removal causes internalization of the receptors and dendritic spines loss. To confirm this hypothesis, we synthesized a cell-permeable peptide, drawn on the PSD95 portion involved in the interaction with JNK (309-REPRRIVI-316 in the PDZ domain) and we demonstrated that this peptide, by inhibiting JNK-PSD95 binding, is able to stabilize PSD95 on the postsynaptic membrane as in control conditions as in pathological conditions, preventing spines degeneration induced by Aß oligomers. The obtained results are the basis for the development of new molecules able to prevent the synaptic dysfunction. Sinaptopathology in animal models suffering of tauopathy In collaboration with a research group of Unimi, we decided to investigate sinaptopathy in a mouse model of neurodegeneration, P301L mice, which is able to replicate one of the characteristic hallmarks of neurodegenerative diseases, Tau protein hyperphosphorylation. This event generates neurofibrillary tangles, causing an alteration of axonal transport, synaptic damage, neuronal death and consequently cognitive and behavioral deficits. Data obtained by the researcher team that featured the model P301L (work submissive PLoS ONE on December 2015), showed a significant interaction between sex and disease, that had never been described in literature. TG females, in fact, have shown higher P-tau levels in cortex and hippocampus than TG males. Consequently, they developed more pronounced cognitive impairment, increased inflammatory damage, with an advanced state of astrogliosis, and increased synaptic damage in these areas. Behavioral tests, such as object recognition test and open field test, were performed to assess behavioral and cognitive deficits in groups of male and female aged 7 and 15 months. This time points corresponds to the first manifestations of the pathological symptoms (7 months) and maximum animal survival (15 months ). The same animals were sacrificed for brain immunohistochemical analysis, evaluating the presence of hyperphosphorylated Tau (AT8 and AT100 isoforms), the inflammatory state (using GFAP) and neuronal damage (using NeuN and Tunel). The data obtained showed not only that the presence of P-tau in cortex and hippocampus is age dependent, but also that it is more pronounced in females than in males, in close dependence with the extended neuronal damage. With these promising data, we decided to investigate the molecular mechanisms underlying neuronal damage and sinaptopathy, analyzing post-synaptic area in male and female mice at 15 months. Results seem to confirm the immunohistochemical data, showing a cortical and hippocampal accumulation of P-tau higher in TG females than TG males. Tau seems localize within dendritic spines, causing severe synaptic damage, postsynaptic receptors loss and to hyperactivation of JNK kinase, a key molecule in neurodegeneration.

Laboratory of Experimental Neurology Role of neuroinflammation in epilepsy In the last 15 years our laboratory carried out pioneer studies on the pathogenic role of neuroinflammation in epilepsy (Vezzani et al, Nature Neurol Rev, 2011), hence our research group is leading investigations related to pro-inflammatory molecules such as interleukin (IL)-1 and High Mobility Group Box 1 (HMGB1). Using experimental models of epilepsy in rats and mice, we demonstrated that epileptogenic injuries evoke neuroinflammatory processes in brain areas involved in seizure generation and spread. These processes include the release of IL-1beta and HMGB1 from glial cells and neurons, and these molecules contribute to seizure generation by decreasing excitability threshold in epileptogenic regions. Then, recurrent seizures and neuropathology trigger further neuroinflammation, thereby perpetuating a vicious pathologic cycle. We are now studying the therapeutic effects of specific anti-inflammatory treatments for developing novel clinical therapies for pharmacoristant epilepsies. We are also addressing the possibility of arresting epilepsy development after an inciting event, or modifying favourably the disease course after its onset, using specific anti-inflammatory drugs either alone or in combination. Finally, we are unraveling the molecular mechanisms mediating the pro-ictogenic activity of some neuroinflammatory mediators, and their involvement in cell loss and epilepsy co-morbidities. This project is part of a EU-FP7 framework programme sponsored study named EPITARGET (2013-2018). Role of Toll-like receptor signaling in epilepsy and neurological sequelae

Infection and fever, which are concomitant with increased levels of pro-inflammatory molecules in the periphery and the brain, can be precipitating events of seizures; moreover, a causal link between CNS infection and epilepsy has been proposed. In the context of convergence of brain infection and the epileptic process, an obvious candidate is represented by the Toll-like receptor (TLRs) family. These receptors are pivotal for the activation of innate immunity and inflammation following either infections or epileptogenic brain injuries (i.e. sterile neuroinflammation). We recently described that the activation of the HMGB1-TLR4 axis lowers excitability threshold, thereby promoting seizure generation (Maroso et al, Nature Med, 2010). We are now studying the role played by TLR3 activation since they sense viral infections (a risk factor for seizures in the pediatric population) and can be activated by endogenous molecules, such as genetic material released by damaged cells. We are studying the molecular mechanisms activated by TLR3 in neurons and astrocytes, and their impact on seizure threshold and cognitive deficits using in vivo murine models and in vitro cell cultures. Characterization and validation of epilepsy biomarkers The development of novel anti-epileptogenic therapies requires the development of non invasive biomarkers of epileptogenesis that could help to identify patients at risk of developing the disease, monitor the disease progression after its diagnosis, and possibly predict the therapeutic response to novel drugs. Our studies focus on the identification and validation of biomarkers in experimental models of epilepsy that could be measurable in blood, or with imaging techniques such as MRI or MR spectroscopy. This project is developed in particular within the Unit of Pathophysiolgy of glio-neuronal communication. Epigenetic control of neuroinflammation in epilepsy: the role of microRNAs

microRNAs (miRNAs) have a key role in post-transcriptional gene regulation of several biological processes in the brain. Specific miRNAs represent a new class of modulators of the inflammatory response in the brain. Our studies focus on specific inflammation-related miRNAs shown to be upregulated in epileptogenic foci both in experimental models as well as in humans. The overall goal is to implement their endogenous levels by ad hoc pharmacological approaches for improving their control of neuroinflammation. This is expected to prevent the deleterious effects of neuroinflammation, thereby mediating anti-ictogenic and anti-epileptogenic actions. Boosting the resolution of neuroinflammation in epilepsy A key role of the brain immune response to pathogens or injuries is to activate homeostatic programmes in immunocompetent cells for tissue defense or repair. This task is achieved by inducing the release of soluble inflammatory mediators acting as effector molecules on target cells. Resolution of inflammation is a highly coordinated and active process that is controlled by endogenous pro-resolving lipid mediators and proteins, and is instrumental to switch off inflammation before it becomes detrimental for tissue. If this mechanism fails then inflammation perpetuates, thereby resulting in tissue injury or dysfunction. A crucial question is how microglia and astrocytes, or leukocytes, balance these tissue demands after injury, and how their behavior can be modified to ameliorate inflammation outcomes. Our hypothesis is that the brain immune response triggered by epileptogenic injuries is inefficiently controlled by pro-resolving endogenous molecules and their cognate G-protein coupled receptors, thus resulting in persistent neuroinflammation. Using experimental models of seizures and post-injury epilepsy, we are studying the role of key pro-resolving molecules such as resolvins and lipoxins, as well as annexins, governing the post-injury inflammatory response. The overarching goal is to implement the brain pro-resolving mechanisms of inflammation for developing new therapeutic strategies. Time-lapse single-cell Ca2+ imaging as a read-out of pathophysiological cell activation This project investigates the neuromodulatory activity of pro-inflammatory mediators and the astrocitic cell response to them by analyzing changes in intracellular Ca2+ signals. These studies use time-lapse single-cell Ca2+ imaging in primary cell cultures from wild-type and mutant mice with impaired cytokine signaling. Bioluminescence and fluorescence functional imaging of IL-1βeta promoter activation

This project is aimed at studying neuroinflammation during epileptogenesis by in vivo bioluminescence and fluorescence functional imaging of IL-1βeta promoter activation. Bioluminescence or fluorescence can be recorded in the brain at multiple individual days during epileptogenesis in the same alive animal. The approach allows to study the time-course and extent of IL-1 promoter activity, its dependence on the seizure activity in the chronic phase, and its modulation by pharmacological interventions for therapeutic purposes. Laboratory of Geriatric Neuropsychiatry Prospective population study on the dementias in the oldest-old Parallel to the progressive increase of individuals aged 80 years or older within the elderly population (65+), the number of demented patients of 80 years or older makes up an ever increasing fraction of the total population affected by dementia. As very often happens, the exclusion from studies of subjects in the oldest age classes tends to inevitably underestimate the total number of individuals affected by dementia present in the population. To fill this gap, a door-to-door population study on the prevalence, incidence, risk factors and evolution of dementias and age-associated cognitive deficits has been set up in an elderly population aged 80 years or older living in eight small towns of Varese Province. The survey was subsequently extended to all registered individuals aged 100 or older residing in the province of Varese. The study is funded by a grant from the Fondazione Italo Monzino, Milano. Health and Anemia in the elderly population

A large survey in old residents of Biella (65 years or older) has been conducted in collaboration with the Local Health Authority of Biella (ASL 12) and with the Division of Hematology, University of Pavia and Fondazione IRCCS Policlinico S. Matteo, Pavia, to estimate the prevalence and incidence of anemia (mild, moderate and severe) in the elderly population and to investigate whether low hemoglobin concentration associated to alteration of CBC such as mean corpuscular volume, leukocytes and/or platatelet cell counts could predict or were associated with myelodysplastic syndrome in the elderly. Prevalence of chronic kidney disease increases considerably with age but little is known about its clinical significance in the oldest old. We have investigated the association of all-cause mortality with a reduced glomerular filtration rate estimated using five commonly used equations [Cockcroft-Gault (C-G), Modification of Diet in Renal Disease (MDRD), MAYO Clinic quadratic equation, Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI), Berlin Initiative Study-1 (BIS-1)] in 700 subjects aged 85 and older taking part in the “Health and Anemia” Study. We have also investigated in this population of oldest old the cross-sectional association between reduced kidney function (estimated as glomerular filtration rate using the MDRD formula) and cognitive performance (assessed with different neuropsychologic tests)

Evaluating risk profiles in ambulatory and hospitalised elderly subjects

In collaboration with the Geriatric Division of the Ospedali Regionali of Lugano and Mendrisio, Switzerland, hospitalized and ambulatory patients are evaluated from a neuropsychological, functional and mobility point of view to estimate the impact of these factors on heath-related outcomes and disease progression (Canton Ticino Study).

Longitudinal study of individuals with mild cognitive impairment (MCI)

In collaboration with the Geriatric Unit of Ospedali Regionali of Lugano and Mendrisio, Switzerland, the follow-up study of all Mild Cognitive Impairment or Questionable Dementia (CDR 0.5) patients seen at the Memory Clinic of the Hospitals is continuing to estimate the rate of conversion to dementia and to evaluate the possible risk factors associated with conversion (Canton Ticino Study).

The Centenari a Trieste Project (CaT): Study of Cognitive, Biological and Social Features of the Population of Centenarians in Trieste and construction of a data-base

Centenarians agreeing to partecipate in the study were interviewed about past and present life styles and evaluated for the presence of disability, depressive symptoms and, specifically, symptoms of cognitive impairment. The medical history and medication are being provided mainly by querying the Local Health Unit’s administrative databases. Sleep activity level and heart rate variability will also be investigated. Finally the consenting individuals were asked to donate a few milliliters of venous blood to perform genetic, epigenetic and proteomics determinations.

A European Multicentre Double-Blind Placebo Controlled trial of Nilvadipine in Mild to Moderate Alzheimer’s disease (NILVAD Project - European Union FP7 Program) In collaboration with the Trinity College Dublin and St. James’s Hospital Dublin together with other ten centres from eight European countries participating in the NILVAD Project. The study employs a randomized double-blind placebo controlled parallel design. The objectives of this study are to investigate the efficacy and safety of Nilvadipine (8 mg once a day) as a disease course modifying treatment for mild to moderate Alzheimer’s disease in a phase III double-blind placebo-controlled study. The primary efficacy outcome measures in this study is the change from baseline to week 78 in cognitive function, as assessed by the Alzheimer’s -Disease Assessment Scale (ADAS -Cog 12). A total of 500 subjects over age 50 years with mild to moderate Alzheimer’s disease (NINCDS-ADRDA criteria); 250 in the nilvadipine group and 250 in the placebo group. The total study duration will be 82 weeks. Patients will receive study medication for 78 weeks. Fatal Familial Insomnia (FFI): preventive treatment with doxycycline of at risk individuals

Department of Neuroscience, in collaboration with 3nd Department of Internal Medicine, Medicine Operative, Unit Oderzo − ASL 9 Treviso and with Fondazione IRCCS Istituto Neurologico "Carlo Besta"The objective of this study is to test whether the chronic administration of 100 mg of doxycycline can prevent (or postpone) the onset of FFI in members of a family carrying the genetic mutation of the prion protein. Survival of the treated individuals will be evaluated after 11 years. Analyses of health data taken from linked administrative databases Following the establishment of administrative databases to monitor medical expenditure reimbursed by the National Health Service, a new field is open to study health using indirect data coming from these sources. We are actively collaborating in analysing data on old subjects and patients with dementia related problems. Quality of care of terminally ill oncological subjects

In 2000 we started a collaborative programme with the hospice “via di Natale Franco Gallini” in Aviano (PN). The present aim of the collaborative research project is to investigate both the clinical and sociodemographic determinants associated with awareness of illness severity in a cohort of terminal cancer patients (n=1080) at the time of admission to the hospice, from 2001 to 2011. In addition we have studied the prescription frequency of pain killer drugs to oncologic terminally patients, in particular analgesics and oppioids, and investigated the prescription trends between 2001 and 2011. Laboratory of Inflammation and Nervous System Diseases The complement system in stroke and traumatic brain injury experimental models Previous studies of ours have indicated that the complement system may represent a novel target for reducing damage following acute brain injury. We showed that C1-INH, an endogenous inhibitor of the complement system currently used in humans as replacement therapy for angioedema, protects against brain injury with a wide therapeutic window. Our data strongly suggested that this remarkable property of C1-INH was due to its ability to bind mannose-binding lectin (MBL), a key protein of the complement lectin pathway. Consistently we later showed that MBL pharmacological inhibition reduces functional and anatomical damage with a wide time window of efficacy (up to 18-24h) in experimental mouse models of

ischemia. Ongoing studies show that MBL is deposited on the ischemic endothelium eliciting its toxic effect. Our present aim is to clarify the pathogenic mechanism driven by MBL on the ischemic endothelium focussing on the interplay between the lectin pathway and the ischemia-induced pro-coagulant endothelium. MBL presence has been demonstrated also after TBI, in human and mouse injured brain where its deletion is protective. Ongoing studies evaluate whether MBL inhibition could represent a new therapeutic strategy also for TBI. In patients we have recently shown that the activation of lectin pathway reflects the severity of brain injury after subarachnoid hemorrhage, a stroke subtype due to aneurismal rupture. Ongoing studies are aimed at assessing the lectin pathway activation in different stroke subtypes, longitudinally, in relation to injury severity, progression and occurrence of complications and to identify genetic factors contributing to interindividual differences. Morphology-phenotype-function relationship in microglia after brain injury Microglia, the major cellular contributors to post-injury inflammation, have the potential to act as markers of disease onset and progression and to contribute to neurological outcome of brain trauma or stroke. After acute injury, these resident cells are rapidly activated and undergo dramatic morphological and phenotypic changes. This intrinsic response is associated to recruitment of blood-born macrophages which migrate into the injured brain parenchyma. We have explored the ability of microglia and recruited macrophages (M/M), to affect neuronal function and promote neurotoxicity through the expression of several harmful components such as inflammatory cytokines, proteases, reactive oxygen and nitrogen species as well as through the interaction with other inflammatory systems such as the complement system. On the other hand we have also documented that these cells possess protective qualities and may promote neurogenesis and lesion repair. M/M are in fact involved in injury resolution and ristorative processes due to the dynamic properties of these cells that are able to rapidly change their phenotype according to the requirment of the injured tissue. The different activation states are characterized by a specific pattern of morphological changes and acquisition of phenotypic markers, whose expression depends on the temporal evolution and on the nature of the brain lesion. Our ongoing studies are aimed at getting insight on previously unexplored aspects of M/M phenotype changes induced by acute brain injury, namely, the morphology and dynamics of activated cells, the presence of specific phenotype markers, whether they are expressed at distinct phases or locations within the lesion, whether they co-label with some complement factors. A deeper knowledge of the M/M features will allow to properly manipulate the inflammatory response to promote a protective environment for therapeutic purposes.   

In vivo real time imaging in ischemic mouse brain by two-photon microscopy Biological events occur in a physical space within a specific time frame and require the action of solid objects such as blood vessels and cells, which interact in a complex network. Information on spatial motility, time-dependent dynamics and tissue integrity (e.g. simultaneous presence of all the cell types involved in the biological effect) is lost with conventional biochemical, biomolecular or histological techniques. In neurobiology in vivo two-photon microscopy offers a new way to tackle previously unexplored mechanisms in physiological or pathological conditions, thanks to its ability to provide a three-dimensional, high-resolution representation of the brain over time in living animals. We have developed in vivo two-photon microscopy protocols to image and quantify brain dynamics after cerebral ischemia and to map the cerebral distribution of injected molecules. We measured the vascular changes that ischemia induces such as the drop in blood flow velocity and flux rate, the occurrence of vessel leakage and the changes in vessel architecture. We used these parameters to study the vascular effects of protective manipulations targeting secondary thrombosis after ischemia. We have visualized immune cells that activate in the ischemic territory, e.g. T-cells and microglia and measured their dynamic behaviour, including cell motility, ramification dynamics and morphological changes after injury. We have also applied in vivo two-photon microscopy to follow brain distribution of fluo-labeled nanoparticles injected systemically, showing their long-lasting persitence with no extravasation. Understanding brain dynamics after injury as well as mapping cerebral drug distribution is aimed at the development of new effective therapeutic strategies for treating brain diseases.  

Understanding the mechanisms involved in the stem cell induced protection

We have provided evidence that mesenchymal stromal cells (MSC) stimulate protective and restorative processes through the secretion of bioactive factors, indicating the potential for a cell free approach. We are now investigating the efficacy of MSC derived secretome to understand whether it may represent a therapeutic strategy instead of cell therapy. Specifically we aim at: i) capturing the MSC-derived key effectors that induce protection after acute brain injury; ii) performing the synthetic reconstruction of the identified neuro-protective cocktail; iii) providing mechanistic insight onto how MSC derivatives affect systemic and brain cell populations. Definition of a successful protocol to be translated to the clinical setting We have shown the MSC efficacy after an immunological mismatched transplant (human MSC in injured animals). These data provide evidence of the possibility of an allogenic transplant in which donor and host are different individuals, allowing a rapid treatment after acute brain injury. These results are an important step towards translation to clinical practice. We are now analyzing the peculiar features of MSC isolated from different sources (bone marrow, cord blood, amnion, chorionic villi, adipose tissue). As stromal cells, they hold common traits, but they also posses peculiarities of their niche that could make a specific MSC source more or less prone to fulfil specific clinical needs. Finally, epidemiological data show a pick of stroke and TBI incidence in the elderly. We are now studying the MSC efficacy in the aged mice to provide evidence of efficacy in this specific population.

Laboratory of Molecular Neurobiology Study of the mechanisms governing the pathogenesis and the course of the Amyotrophic Lateral Sclerosis Comparative analysis of two murine models of familial ALS with phenotypic differences of disease for the identification of prognostic markers and therapeutic targets. Continue the comparative study between the two mouse models of ALS carrying the SOD1G93A mutation and showing a different disease phenotype in terms of disease onset and duration due to the different genetic background. Having further highlighted that the rapid course of the disease depends on an earlier denervation of neuromuscular junctions rather than from a greater loss of motor neurons, we focus the attention on the study of the peripheral nerves and neuromuscular junctions in these two mouse models. We have observed that at the onset of the disease, the sciatic nerves of mice fast progressor show low levels of NF200 and of acetylated tubulin, two markers of structural functionality of the axon, while this is not observed in mice slow progressing. On the contrary, infiltrates of macrophages and CD8 + T cells in the nerves are significantly higher in mice with a more benign course of the disease than those with the most aggressive form. This reflects increased levels of CCL2, complement C3 and MHCI that are much higher in motor axons of mice with slower disease. Therefore, these observations lead us to speculate that the preservation of the peripheral nervous system through a positive immune response mediated by CCL2, complement 3 and MHCI have a positive impact on the disease. We are therefore examining in more detail the role of the immune system to identify common targets on which to design new therapies. Below is a description of these studies. Also continues the collaboration with Dr. Malaspina of Queen Mary University of London for the analysis of various tissues isolated from two ALS mouse models with a system of proteome analysis very sensitive and innovative in order to identify the molecular signals in the blood, which can be indicators of disease prognosis as well as potential therapeutic targets. (Project funded by MNDA U. K. and EUROMOTOR FP7 program). Role of neuroinflammation and neuroimmunity in the pathogenesis and progression of ALS. Our recent studies and other evidence indicate that the involvement of the immune system in ALS is no longer a secondary event following the progressive degeneration of motor neurons but rather it can be a causative phenomenon in governing the development and in particular the severity of the ALS. Our interest in this regard is aimed to the study of the following mechanisms:

Pathogenic role and possible clinical use of the axis CCL2/CCR2 in the regulation of immune responses in ALS. This translational research project aims to understand the role of the axis CCL2/CCR2 in the induction and maintenance of the immune response in ALS. The project is based on the assumption that an early increase of CCL2/MCP-1 expression as found in the spinal cord of SOD1G93A mice and presumably in ALS patients, can attract monocytes and lymphocytes from the blood able to exert a potential protective effect on damaged motoneuron. The results in ALS patients (n = 73) versus healthy controls (n = 43) showed that in ALS patients the framework of the immune system in peripheral blood is quite variable and index of inflammation as evidenced by the intense and prolonged CD14 inflammatory monocytes and their levels of CCR2 receptor and by the decrease in CD16. We also find that the levels of Treg were significantly lower in ALS patients compared to healthy control in line with previous observations. In SOD1G93A mice there was no evidence of changes in inflammatory monocyte and their CCR2 receptor in peripheral blood. However, the study of CCL2 / CCR2 axis in the central and peripheral nervous system has shown that the motor neurons of SOD1G93A mice at the onset of symptoms, express high levels of CCL2 , and this is accompanied by the activation of microglia / macrophage but not monocytes. Increased CCL2 is also observed in the motor axons of sciatic nerve and is accompanied by macrophage infiltration. Since this phenomenon is prevalent in mice with a more begnin disease we hypothesize that this axis is activated to promote a protective immune response. In support of this hypothesis, we observed that in primary cultures of motor neurons the inhibition of CCR2 receptors is neurotoxic in a dose dependent manner. We also tried to exploit the axis CCL2 / CCR2 to favour the recruitment of protective Treg cells, suitably transfected with the CCR2 receptor, in the spinal cord of SOD1G93A mice and to assess the impact on disease progression. Unfortunately, the preliminary result showed that these cells are unable to migrate in the spinal cord when transplanted intracerebroventricularly and they are not recruited in the peripheral nerves when injected intravenously. We envisage to consider other strategies to favour the homing of these protective cells. This project supported by the Italian Agency for the research on ALS (ARISLA) is based on collaboration between our group, the Department of Immunology of Humanitas Foundation for the research of Milan and the Fondazione Salvatore Maugeri IRCCS, Scientific Institute in Milan. Role of The Major Histocompatibility Complex I (MHC-I) In Amyotrophic Lateral Sclerosis In order to understand the role that the MHC-I plays in the pathogenesis and progression of the disease in mice SOD1G93A, we have crossed them with mice lacking beta-2 microglobulin (b2m), an essential component in the function of MHCI and the adaptive immune response. After several crossings we got SOD1G93A mice lacking the protein b2m (SOD1G93A / b2m-/-) in which we analyzed the performance of neuromuscular deficits, histopathology and the adaptive immune response compared with mice with normal levels of SOD1G93A b2m (SOD1G93A / b2m+/+). The data show that SOD1G93A / b2m-/- have a significant anticipation in the onset of motor deficit than mice SOD1G93A / b2m+/+. This phenomenon correlates with a more precocious denervation and muscles atrophy of the hind legs and with more pronounced structural changes of motor axons in the peripheral nerves. In conjunction with these changes, it was found a reduced recruitment of immune cells (macrophages and CD8Tcells) in peripheral nerves and a reduced proliferation of Schwann cells (SCs), a phenomenon associated with axonal stress. Therefore, these results demonstrate that an adaptive immune response mediated by the activation of MHCI at the level of the peripheral nerves and neuromuscular junctions is critical to maintaining the neuromuscular activity thus favoring a more benign disease. On the contrary, the motor neurons in the lumbar spinal cord are more protected in SOD1G93A / b2m-/- mice, an unexpected effect which is under examination. In general, however, this study again demonstrates that the preservation of the motor neuron is by itself a phenomenon insufficient to halt the progression and to improve the clinical course of the disease. Furthermore, this study emphasizes the importance of maintaining the activated complex MHCI in peripheral nerve in order to maintain as long as possible a functional innervation and slow muscle atrophy. The manuscript with these findings has been submitted to the journal Brain for publication. This project is funded by the Foundation Thierry Latran for the Research on ALS from France and is made in collaboration with the Department of neuroscience of the Karolinska Institute in Stocholm, Sweden.

Role of the chemokine CXCL13 in ALS

Comparative transcriptome analysis conducted on motor neurons of the two mouse models of ALS with different clinical phenotypes, showed that in mice with rapid disease progression the increased expression of the chemokine CXCL13 was much more pronounced than in mice with slow disease progression already from the pre-symptomatic stage. These data were confirmed in terms of protein levels in the lumbar spinal cord with a preferntial expression in motor neurons and microglia and in cerebrospinal fluid (CSF). CXCL13, which is a chemoattractant of B cells, is known to be constitutively expressed in dendritic cells of the secondary lymphoid organs where regulates the structural and functional organization of the follicle and in some lymphocyte subtypes of type T. The specific receptor through which it acts, CXCR5 , is expressed in mature B cells, follicular T helper cells and also pro-inflammatory Th17 cells that produce IL-17. Increased levels of IL-17 were observed in the lumbar spinal cord of SOD1G93A mice particularly in those with rapid disease course, as well as in the central nervous system of ALS patients. Therefore we hypothesise that an aberrant recruitment of Th17 cells, could help the triggering of the inflammatory process. Currently studies are underway to investigate this hypothesis and to understand the role of CXCL13 in the pathogenesis of ALS.

For this study we established a collaboration with the company Vaccinex (Rochester, NY, USA) which kindly provided us the CXCL13 antibody and the IgG control to assess the effect of chronic treatment on the disease.

Effect of RNS60, an anti-inflammatory agent in in vitro and in vivo models of ALS. RNS60 is a fluid composed of saline and oxygen subjected to an electrokinetic processing to produce charge-stabilized nanostructures (CSNs). The CSNs consist of an oxygen core surrounded by layers of positive and negative electrical charges. In various experimental models both in vitro and in vivo RNS60 has been shown to reduce activated microglia, inhibit neuroinflammation and exhinit neuroprotective properties. In addition, RNS60 was reported to increase the protective Treg in a mouse model of experimental allergic encephalomyelitis (EAE), and this was accompanied by a significant improvemente of the disease course. Based on this evidence, we first evaluated the neuroprotective and anti-inflammatory RNS60 in co-cultures of astrocytes and spinal neurons from transgenic mice SOD1G93A. We found that RNS60 is able to protect motor neurons from death induced by the presence of the mutated SOD1 and to reduce the levels of iNOS produced by astrocytes. We then evaluated the effect of RNS60 on the disease course of SOD1G93A mice. RNS60 treatment was able to delay the onset of motor symptoms and to modestly increase the survival of mice. Further studies are underway to validate these preliminary results and to analyse the mechanism of action of RNS60. The RNS60 is a product of Revalesio Corporation Tacoma (Washington), which funded the project. Effect of repeated treatment with human umbilical stem cells (UC-MSCs) into mice with familial ALS. The objective of this study is to evaluate whether repeated administration of human umbilical mesenchymal stem cells have a long-term effectiveness in improving the performance of the disease and increase survival in transgenic SOD1G93A mice and whether this correlated with their immunomodulatory and cytoprotective effect. Mesenchymal stem cells from the human umbilical cord wall was provided by the laboratory of Cell Therapy "G.Lanzani" of Bergamo. In the first part of the study we assessed the distribution of the cells in the brain and spinal cord after intravenous or intracerebroventricular administration in order to optimize the protocol for a better efficacy. Therefore the cells were labeled with fluorescent nanoparticles provided by the Politecnico of Milan before being injected and then followed by analysis in vivo through whole body fluorescence scanning and ex vivo by the histology of the nerve tissue, in collaboration with the Unit of the Department of Biochemistry Nanobiology Mario Negri Institute. Based on the results we decided to administer the cells into the cerebral ventricles of mice SOD1G93A every 2 weeks from onset of symptoms, to a total of 4 doses in order to evaluate their effectiveness on the disease course.The results show that there is no effect on survival although there is a partial protection of motor neurons, associated with the reduction of activated microglia, in the lumbar spinal cord. Analyses are in progress to test the degree of

denervation of muscle junctions in order to determine whether the lack of effect on survival is due to the lack of protection in this district.

Laboratory of Experimental Psychopharmacology Drug Abuse: Neural basis of drug self-administration To separate the direct pharmacological effects of cocaine from those associated with active drug self-administration we employed a yoked control-operant paradigm and investigated the expression of well established markers of the rapid action of cocaine, i.e. the inducible early genes, such as Activity-Regulated Cytoskeletal-associated protein (Arc), and trophic factors, such as Brain Derived Neurotrophic Factor (BDNF) and alpha calcium/calmodulin-dependent protein kinase type II (αCaMKII), in rats after a intravenous (i.v.) cocaine self-administration session. Animals self-administering cocaine (SA) did more active lever-presses than yoked-cocaine (YC) and yoked-vehicle (YV) animals. This goal-oriented behaviour was accompanied by a selective increase in Arc mRNA levels in the medial prefrontal cortex (mPFC). These findings demonstrate that a single session of cocaine i.v. self-administration is sufficient to shape rat behaviour towards goal-directed behaviours and selectively up-regulate Arc expression in mPFC (of SA animals), providing the first evidence that the mPFC's function is already profoundly influenced by the first voluntary cocaine exposure. Ongoing studies are evaluating whether this effect is peculiar to cocaine or common to other drugs of abuse. BDNF dynamic changes were investigated in the nucleus accumbens (NAc) and mPFC during use and the early phases of cocaine abstinence after chronic exposure by employing a “yoked control-operant paradigm”. The effect on BDNF was region-specific and dependent on the withdrawal time. In the NAc, BDNF protein levels increased immediately after the last self-administration session, with a larger increase in passively cocaine-exposed rats. In the mPFC, BDNF expression was elevated 24 hours after the last self-administration session, independently of how the drug was encountered. No changes were found in NAc and mPFC 7 days after the last self-administration session. Analysis of transcript levels in the mPFC indicated that action on exon I might contribute to BDNF's cortical induction. Increases in alpha calcium/calmodulin-dependent protein kinase type II (αCaMKII) activity in the nucleus accumbens shell has been proposed as a core component in the motivation to self-administer cocaine and in priming-induced drug-seeking. Since cocaine withdrawal promotes drug-seeking, we hypothesized that abstinence from cocaine self-administration should enhance αCaMKII as well. Short-term abstinence from contingent, but not non-contingent, cocaine i.v. self-administration elevates αCaMKII autophosphorylation, but not the kinase expression, in a dynamic, time- and brain region-dependent manner. Increased αCaMKII autophosphorylation in the nucleus accumbens (NAc) and medial prefrontal cortex (mPFC), but not dorsolateral striatum (dlS), was found 24 h, but not immediately, after the last cocaine self-administration session. Notably, in the mPFC, but not NAc and dlS, αCaMKII autophosphorylation was still enhanced 7 d later.

Neural basis of “drug craving” and “relapse” in the drug abuse assumption Drug craving, defined as the desire to experience the effect(s) of a previously experienced psychoactive substance is a cardinal feature of drug addiction and is clinically significant because of its potential link to relapse. To provide useful indications to the development of novel therapeutic approaches to prevent the use and abuse and the relapse of drug assumption following the outcome of craving, we elaborated experimental models of self-administration and relapse induced by cocaine, nicotine and alcohol-associated cues, after a period of abstinence. Ongoing studies are evaluating the role of several neurochemical mechanisms potentially involved in the drug-seeking behaviour.

Search for pharmacological agents modulating drug craving and relapse Environmental stimuli associated with the intake of psychotropic substances of abuse may have the ability to induce the craving that often preludes to relapse in formally detoxified patients. Studying nicotine in an experimental model of extinction-reinstatement induced by the presentation of environmental stimuli associated with self-administration of psychotropic substance of abuse, it was found that bifeprunox, a high-affinity partial agonist of dopamine (DA) D2 receptors and serotonin1A (5-HT1A) receptors, preferentially reduced nicotine-seeking behaviour in response to drug-associated stimuli in rats after a long period of abstinence. Pharmacological stimulation of N-methyl-D-aspartate receptors (NMDAr) could enhance the outcome of cue-exposure therapy for smoking cessation. NMDAr stimulation can be achieved by increasing

pharmacologically the synaptic levels of glycine, a necessary co-agonist. Here, we evaluate the effects of SSR504734, a selective inhibitor of glycine type I transporter (GlyT1) in an extinction-reinstatement procedure inducing robust and lasting nicotine-seeking behavior in rats. Acute pre-treatment with SSR504734 reduced nicotine-seeking but not sucrose-seeking behavior without influencing rats' locomotor activity. Sub-chronic treatment during daily exposure to nicotine-conditioned cue reduced nicotine-seeking; however, this effect was transient, with return to responding at 72 hours. Full recovery of responding was observed after 1 month suggesting that SSR504734 sub-acute treatment did not engage the long-term plasticity mechanisms probably involved in nicotine-seeking. Laboratory of Epidemiology and Social Psychiatry Development of a Manual of Family therapy based on the systemic model integrated with the attachment theory for adoptive families with problematic adoptive children In order to standardize the phases of the family therapy, 23 sessions, covering all the phases, were observed and the phases identified. Concordance indices were computed, the manual revised accordingly and a fidelity scale was developed. Validation of the Italian versione of the Health of the Nation Ouitcome Scales Children and Adolescents, HoNOS-CA In order to introduce the HoNOS-CA in the routine activity of Child Psychiatry services the reproducibility of the scale was assessed and the collection of data for the validation process is ongoing. Ten services have collected data on 134 cases, comparing the HoNOS-Ca with the Strenghts and Difficulties Questionnaire, filled by the caregiver, and the CGI. Data were collected for each subject after four months too, in order to assess the responsiveness of the tool. Integrated interventions to promote early identification and treatment of severe mental illnesses in the young (15-24 yars) and in at risk population groups In the framework of psychoses prevention, the project addresses ultra high risk mental states, or prodromes of psychotic crisis. Ultra high risk mental states have a transition to psychosis rate ranging between 10 and 40%, which can be halved by appropriate intervention. Psychosis is more common in migrants, where access to services and identification of psychosis and symtoms may be more difficult. Mental health services are now trying to have a more integrated organization of the necessary knowledge and competencies. The Laboratory is involved in developing and disseminating such competencies and defining appropriate interventions offered according to severity. FOCUS - Frailty management Optimisation though EIP AHA Commitments and Utilisation of Stakeholders input In order to critically reduce the burden of frailty in Europe the project assists partners in better grounding prevention and management of frailty into real needs, expectations and experiences of all stakeholders. A meta-synthesis of qualitative studies about the conceptialization of frailty and experiences of care and prevention was conducted and five focus groups with healthy older individuals, frail older individuals, caregivers, health and social professionals were held.

Laboratory Prion Neurobiology Prion diseases Prion diseases, also known as transmissible spongiform encephalopathies, are progressive and invariably fatal degenerative disorders of the central nervous system that affect humans and other animals. Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker (GSS) syndrome, and fatal familial insomnia (FFI) are the most common forms in humans; scrapie of the goat and sheep, bovine spongiform encephalopathy (“mad cow disease”), and chronic wasting disease of deer and elk are the best-known examples of prion zoonoses. These diseases result from the conformational change of a cellular protein of unclear function (denominated

prion protein, PrP) into a self-propagating pathogenic isoform that accumulates in the brain of the patients and causes neuronal dysfunction and degeneration through an unknown mechanism. Three different manifestations of prion diseases are recognized: sporadic, infectious and genetic. Genetic prion diseases display autosomal dominant inheritance and are linked to insertional and point mutations in the PrP gene, on chromosome 20. These mutations are presumed to favor the conformational conversion of PrP into a pathogenic isoform. Interestingly, different mutations are associated with different types of prion disease (CJD, GSS or FFI). The research activity in the laboratory of Prion Neurobiology is focused on two main questions: 1) What causes neuronal dysfunction in inherited prion diseases? 2) How do different PrP mutations cause different diseases? We have developed a research program to tackle these questions, using transfected cells, transgenic mice and primary neuronal cultures for complementary exploration of responses to mutant prion proteins. These experimental models are being analyzed with a wide range of molecular and cell biology techniques, as well as protein chemistry and proteomics. The major achievements of the laboratory are the development of the first transgenic mouse model of Creutzfeldt-Jakob disease that recapitulates the cognitive, motor and neurophysiological abnormalities of the human disorder (Dossena et al., Neuron, 2008), and the discovery of the molecular mechanism by which mutant PrP induces neuronal dysfunction (Senatore et al., Neuron, 2012).

Alteration of intracellular trafficking in synaptic dysfunction in genetic prion diseases

Synaptic dysfunction is an important cause of neurological symptoms in prion diseases, a class of clinically heterogeneous neurodegenerative disorders caused by misfolding of the cellular prion protein (PrP). Experimental data suggest that accumulation of misfolded PrP in the endoplasmic reticulum may be crucial in synaptic failure, possibly because of the activation of the translational repression pathway of the unfolded protein response. We report that this pathway is not operative in mouse models of genetic prion disease. Building on our recent finding that intacellular retention of mutant PrP impairs the secretory trafficking of calcium channels essential for synaptic function, we propose a model of pathogenicity in which intracellular retention of misfolded PrP results in loss of function or gain of toxicity of proteins important for synaptic fucntion, such as neurotransmitter receptors and signaling complexes.

Prion protein and voltage-gated calcium channels

We previously found (Senatore et al., Neuron 2012) that PrP interacts with the 2 subunit of voltage-gated calcium channels (VGCCs), suggesting a possible role of PrP in the channel function. We discovered that expression of PrP inhibits the function of VGGCs because it interferes with the GPI-anchoring of the VGCC 2 subunit. This study reveals a new mechanism governing membrane expression of GPI-anchored proteins based on competition for rate-limiting components.

Laboratory of Neurochemistry and Behavior

Role of brain serotonin in the effects of psychostimulants: studies in tryptophan hydroxylase2 ko mice Serotonin (5-HT) is involved in the regulation of many brain functions and in the mechanism of action of various psychotropic drugs including psychostimulants, which motor effects are mainly dependent on the activity of meso-limbic and meso-striatal dopaminergic systems. It is well known that brain serotonergic neurons influence the effects of psychostimulants but their precise contribute remains to be clarified. About ten years ago tryptophan hydroxylase2 (Tph2) was identified as the gene coding for the enzyme responsible for the hydroxylation of tryptophan to form 5-hydroxytryptophan (5-HTP) in the brain, an essential step in the biosynthesis of 5-HT. The generation of Tph2 ko mice permitted to reconsider the role of brain 5-HT in the psychostimulant effect of amphetamine. Amphetamine-induced motor activity is strongly enhanced in Tph2 ko mice and the administration of 5-HTP restores the normal response to amphetamine. Lack of brain 5-HT does not affect the normal development of 5-HT neurons or the synaptic availability of striatal

dopamine in response to amphetamine. The results indicate a significant role of 5-HT in the individual response to amphetamine’s effects. Ongoing studies aim to assess the role of Tph2 on spontaneous activity.

Role of cholesterol metabolism in Rett syndrome: studies in patients and in a mouse model of the pathology Rett syndrome (RTT) is a severe and rare disease caused by mutations of the X-linked methyl-CpG-binding protein2 (Mecp2) gene, a regulator of gene transcription. No disease-modifying therapies are available yet. Recently, it has been shown that Mecp2 deletion affects cholesterol metabolism in mice and suppression of cholesterol synthesis with statins improved survival and motor deficits. Given the pleiotropic effect of statins, it is not clear whether their effect involves cholesterol-dependent or independent mechanisms. In collaboration with Analytical Instrumentation Unit of our institute (R. Bagnati) a HPLC-Electrospray-Mass spectrometry method has been set up to determine plasma levels of 25- hydroxycholesterol (25-OHC) and 24-OHC, a metabolite mainly derived from brain cholesterol and a useful index of brain cholesterol metabolism. Through a collaboration with (Prof. Renieri) and the Telethon biobank we obtained plasma samples of RTT girls where we found alterations of 24-OHC, 25-OHC and their ratio. These preliminary findings will be confirmed on a larger sample that will be available through the collaboration to a clinical study coordinated by Siena Hospital aimed at assessing the potential benefit of statins in RTT. To assess the role of oxysterols, studies in Mecp2 ko mice, an experimental model recapitulating most of the symptoms of RTT, are ongoing.

Laboratory of Neurological Disorders Epidemiological studies on amyotrophic lateral sclerosis (ALS) Included are studies on the incidence, risk factors and mortality of ALS. The data are obtained from a regional registry of the disease activated in 1998 and including all patients with newly diagnosed ALS identified in the Lombardy region. Using similar study protocols, the same data are collected in three additional regional registries (from Piemonte, Liguria and Puglia) included in a network with the Lombard registry. Information obtained from patients enrolled in the Lombard registry and from cases examined by members of the Italian ALS Study Group has been used to assess the validity and reliability of diagnostic criteria for ALS and selected disability scales. Based on the data recorded, the annual incidence of ALS is comparable to that obtained in other Western countries where ALS registries have been activated, and is among the highest ever published (1.9 per 100,000). In contrast, new data have been provided on long-term serviva of ALS. In a color of 483 patients from the Lombard registry, the cumulative time-dependent survival at 1, 5 and 10 years was 76, 23 and 12% respectively. In addition, in men with ALS aged 75 years or older, survival overlapped that of the general population. The study on the validation of the current diagnostic criteria for ALS (the El Escorial criteria) showed that, to be considered valid and reliable, the criteria should be used after proper training of the investigators. In October 2004, the Laboratory of Neurological Disorders has started a European collaborative group for the ALS registries (EURALS) with the intent to create a common database (completed in the year 2005) with the participation of the existing regional and national disease registries. With the collaboration of the UK and Irish groups participating in the EURALS collaboration, a scientific report has been published on a meta-analysis of the incidence of ALS, performed by pooling data from the 1998-99 cohorts of patients enrolled in the population-based registries. Two studies have been recently concluded: 1. A case-control study on trauma and risk of ALS (in collaboration with the Italian registries); 2. A survey of the prevalence of cognitive impairment and extrapyramidal signs in patients with newly diagnosed ALS (Italian registries); 3. A study on the correlation between ALS and coffee intake; 4. A comparative study of the genotype and phenotype of early onset and late onset ALS; 5. A case-control study of sport, physical activity, and trauma and risk of ALS (in collaboration with partners of the EURALS group); 6. A study on the long-term survival of ALS. The following investigations are still in progress: 1- A study in a population-based incident cohort 1998-2002, aimed to verify the correctness of the diagnosis during follow-up; 2. A study comparing cognitive impairment and extrapyramidal signs in a sample of ALS patients and in a matched control population in Lombardy; 3. An observational study to identify environmental and and genetic risk factors in

some European populations. 4. A survey on dietary factors in patients with ALS and healthy controls to investigate the effects of alimentary habits on the disease risk; 5. A systematic review of the literature on the association between ethnic origin and ALS phenotype, incidence and prognosis. Therapeutic trials in neurological disorders During the year 2014-2015 two therapeutic trials sponsored by the Italian Drug Agency (AIFA) and a therapeutic trial sponsored by the Italian Ministry of Health were ongoing. Included are: 1. A randomized open-label trial of the efficacy of a comprehensive rehabilitation program for the prevention of falls in Parkinson’s disease; 2. A randomized open-label trial on the efficacy of an active monitoring of the adverse effects of antiepileptic drugs and of relevant drug interactions.The first trial aims at assessing whether a comprehensive rehabilitation program compared to usual care is followed by a reduction in the incidence of falls in patients with Parkinson’s disease at risk of falls. The second trial aims at verifying the added value of an active monitoring of adverse drug interactions compared to usual care in patients receiving antiepileptic drugs associated to other compounds. The laboratory of neurological disorders is a partner in these trials, where the main tasks include protocol and CRF preparation, statistical analysis, and preparation of the final scientific report. During 2016, a new therapeutic trial will start on the rapid versus slow withdrawal of antiepileptic drugs in two-year seizure-free adult patients with epilepsy. Impact of epilepsy on the quality of life of children and adolescents and their families 345 relatives of children and adolescents with epilepsy were interviewed to verify the impact of the disease on the family’s quality of life. Disease severity and the degree of parents’ concerns correlated with a deterioration of quality of life. In all its aspects even though, compared to their families, adolescents had a more positive attitude towards the disease. Prevalence and incidence of epilepsy (and drug-resistant epilepsy) in northern Italy The study aim was to calculate the prevalence and incidence of epilepsy in a well-defined area of Lombardy, using administrative data for the period 2000-2008 provided by the regional database. Included were patients fulfilling the ICD 9 code for epilepsy and seizures and/or the disability exemption code for epilepsy, the presence of EEG, and antiepileptic drugs prescriptions in variable combinations. The validity of the diagnostic criteria was assessed examining a sample of patients with epilepsy through their caring physicians. The best and most conservative algorithm included EEG and selected treatment schedules (sensitivity 85.9%; specificity 99.8%; positive and negative predictive values 64.2% and 99.9%). Based on these values, data obtained from administrative records were adjusted to provide prevalence ratios and incidence rates of respectively 4.42 per 1,000 and 47.05 per100,000 per year. These data are comparable to those of accurate epidemiological surveys done in industrialized countries. Subsequently, we calculated the prevalence and the incidence of active epilepsy (patients currently being treated and/or having had at least one seizure in the previous five years) and drug-resistant epilepsy (failure of adequate trials of two tolerated and appropriately chosen antiepileptic drugs, whether as monotherapy or in combination, to achieve sustained seizure freedom) using data collected with the collaboration of general practitioners, pediatricians, child neurologists and neurologists of the study area. The prevalence and incidence of active epilepsy were respectively 4.67 per 1,000 and 28.46 per 100,000/year. The frequency of drug-resistance was 15.6% in the prevalent population and 10.5% among incident cases. The prevalence of drug-resistant epilepsy was 0.7 per 1,000, the incidence was 3.0 per 100.000/year. The long-term prognosis of epilepsy was analyzed using four distinct patterns: 1. Remission for at least two years in the first two years of treatment (early remission); 2. Remission after more than two years (late remission); 3. periods of remission followed by relapse; 4. Never remission. 747 subjects of all ages were included. The probability of entering remission for at least two years was 18% at the beginning of treatment and rose to 67% at 20 years. Early remission was seen in 101 people (19%), late remission in 175 (33%), remission followed by relapse in 85 (16%), and no remission in 166 (32%). Among the drug-resistant patients remissions were observed in 30% of cases, of which 13% without relapses. Cochrane Review on immediate antiepileptic drug treatment for first unprovoked seizure, versus deferred, no treatment or placebo.

There is considerable disagreement about the risk of recurrence following a first unprovoked epileptic seizure. A decision about whether to start antiepileptic drug treatment following a first seizure should be informed by information on the size of any reduction in risk of future seizures, the impact on long-term seizure remission, and the risk of adverse effects. The probability of seizure recurrence, seizure remission, mortality, and adverse effects of anti-epileptic drug (AED) treatment given immediately after the first seizure compared to controls, in children and adults, were reviewed. The included studies,randomised controlled trials, compared participants receiving immediate antiepileptic treatment versus those receiving deferred treatment, those assigned to placebo, and those untreated. After exclusion of uninformative papers, only six studies (nine reports) were selected for inclusion. For the two largest studies data were available for individual participant meta-analysis. Compared to controls, participants randomised to immediate treatment had a lower probability of relapse at 12 months, a higher probability of an immediate two-year and an immediate five-year remission. Antiepileptic drugs did not affect overall mortality after a first seizure. Compared to deferred treatment, placebo, or no treatment, treatment of the first seizure was associated with a significantly higher risk of adverse events.The quality of the evidence from the included studies was moderate to high for seizure recurrence, remission and mortality outcomes; with high quality evidence reported in the two large studies we were able to analyse individual participant data. Quality of the evidence for adverse events was low, with variable reporting across studies.

Study of psychogenic nonepileptic seizures (PNES)

Aim of this study was to investigate if, when and to what extent visual information contained in a video-recorded event allows experienced epileptologists to predict the diagnosis without the aid of the EEG. Five neurologists actively practicing in Epilepsy Centers in Italy and USA were asked to review 23 videos capturing representative events of 21 unselected consecutive patients admitted for long-term video-EEG monitoring (VEM). They were requested to 1) rate the videos for quality and content, 2) choose among four diagnoses: a) epileptic seizures (ES); b) psychogenic non-epileptic seizures (PNES); c) Other NES (syncope, moVEMent disorder, migraine, etc.); d) “Cannot Say”; 3) explain in their own words the main reasons leading to the diagnosis of choice. All raters predicted the diagnosis correctly in 7/23 videos (all ES or PNES) (30.4%) whereas all raters failed in 5/23 cases (three Other NES, one PNES, one Cannot Say) (21.7%). Interrater agreement was “moderate” for the overall group; “moderate” for ES (k = 0.53); “substantial” for PNES (k = 0.63); “fair” for Other NES (k = 0.21).

EPINET: An international web database for the collection of data in patients with epilepsy An epilepsy patient database that can be accessed via the Internet by neurologists from anywhere in the world. The database was designed to enroll and follow large cohorts of patients with specific epilepsy syndromes, and to facilitate recruitment of patients for investigator-initiated clinical trials. The EpiNet database records physician-derived information regarding seizure type and frequency, epilepsy syndrome, etiology, drug history, and investigations. A validation study was performed to determine variation in epilepsy diagnosis around the world when neurologists are presented with identical case scenarios. The difficulties in diagnosing epilepsy are under recognised. The EpiNet steering group assessed the inter-rater agreement among neurologists in diagnosing epilepsy, seizure type(s) and etiology. Epileptologists and neurologists with an interest in epilepsy were invited to participate in the study between December 2013 to September 2014. 32 case scenarios describing real patients (6 children, 26 adults) with various events were evaluated online. For each patient, participants were asked: to determine how likely it was the patient had epilepsy; if epilepsy, to classify the patients’ seizures using the ILAE 2010 classification; and to determine the etiology, and epilepsy syndrome when appropriate. The “gold standard” diagnosis was determined by the EpiNet steering committee. 21 patients had epileptic seizures, of whom 11 had focal seizures and 8 generalised seizures; in 2 the steering committee was uncertain if seizures were focal or generalised; 11 had non-epileptic events. In 24 cases, information was presented in 2 steps; clinical information was presented in Step 1, and after completion of this step,

neuroimaging and EEG studies were provided in Step 2. Results 322 neurologists commenced the study. 201 participants from 35 countries completed the 32 cases. Full data was available for the 24 cases with 2 steps from 191 investigators. Kappa values for step 1 were: diagnosis of epilepsy = 0.55; seizure type(s) = 0.36; etiology = 0.36. Kappa values increased at Step 2 for diagnosis of epilepsy (0.68), seizure type (0.48) and etiology (0.44). A study on the long-term prognosis of epilepsy (PRO-LONG) The purpose of the PRO-LONG study was to assess the long-term prognosis in patients with newly diagnosed epilepsy using pre-defined prognostic models. Unselected patients with newly diagnosed epilepsywere recruited by 13 epilepsy centers throughout the country and followed for at least 10 years. Baseline clinical and instrumental findings and data on periods of remission (absence of seizures for at least one year) were collected. Patients were classified according to five prognostic models: early remission, late remission, relapsing-remitting course, worsening and no remission. The sample comprised 1006 unselected patients with newly diagnosed epilepsy. The median follow-up was 16.0 years from the time of diagnosis. During follow-up, 91.9% of patients had a remission of at least one year and 44.4% of at least 10 years. 31.1% of patients early or late remission (23.9% of them off treatment at the end of follow-up). The commonest prognostic model was the relapsing-remitting pattern (52.2%), while in 16.7% a worsening pattern (ie remission followed by relapse and no further remission) or no remission ever was observed. The strongest predictors of long-term remission (10 years or more) were fewer seizures before treatment and a lower number of AED exposures. On the contrary, the probability of a worsening or pattern or no remssion ever was associated to a greater number of seizures before treatment, to a longer duration of epilepsy at diagnosis, to epilepsies with structural-metabolic etiology, to an increased number of drugs, and to epilepsy surgery.

Laboratory of Quality Assessment of Geriatric Services Pharmacoepidemiology

The availability of computerized system for the management and care of community-dwelling and in-hospital patients represents an opportunity for developing and implementing new strategy in the field of the evaluation, monitoring and implementation for the appropriateness of drug prescription and the continuity of care. In collaboration with the Health Directorate of Lombardy Region, several Local Health Units and hospitals, a collaborative study has been set up aimed to test, in some critical prescribing fields, the effectiveness of multidisciplinary integrated interventions and educational events in improving the prescribing practice and to implement the utilisation of generic drugs.

A. PHARMACOEPIDEMILOGICAL STUDY USING ADMINISTRATIVE DATABASE

‐ Prevalence and appropriateness of antidepressant use in elderly. Changes in the patterns of antidepressant prescribing from 2000 to 2007 were investigated and put into relation with the rates of depressive disorders in Lombardy. The 1-year prevalence of “AD use” increased dramatically from 2000 to 2007. The greatest shift occurred between 2000 and 2003 when the global prescription almost doubled increasing from 5.5% to 9.9%. The most pronounced increase was seen in females who in 2007 reached a 1-year prevalence of AD use of 13.8%. The prescription of TCAs and other ADs remained stable across the years, thus the observed changes were mainly attributable to SSRIs. The SSRIs accounted for 44.8% of “AD use” in 2000 and rose to 75.7% in 2007. The most prescribed antidepressant was citalopram: its 1-year prevalence increased about sixfold and, in 2007, peaked at 3.3%. Citalopram was followed by two SSRI: paroxetine (2.2%) and sertraline (1.9%).

‐ Antipsychotics prescription and cerebrovascular events in Italian older persons. Administrative claims from community-dwelling people aged 65 to 94 years living in Northern Italy were analysed using a retrospective case-control design, from 2003 to 2005. 3855 cases of cerebrovascular events (CVEs) were identified and matched with 15420 controls. When antipsychotics were categorized according to number of boxes prescribed during the observational period, being prescribed with at least 19 boxes of typical antipsychotics was significantly associated with CVEs (OR=2.4;95%CI=1.08-5.5). An interaction was found between any antipsychotic and acetilcholinesterase inhibitors (AChEI) co-

prescription on CVEs (OR=0.46;95%CI=0.23-0.92). In conclusions, only typical antipsychotics were associated with an increased odd of CVEs but the association was duration-dependent. Persons prescribed simultaneously with AChEI and antipsychotics may be at a lower risk of CVEs.

‐ New prescriptions of spironolactone associated with angiotensin-converting-enzyme inhibitors and/or angiotensin receptor blockers and their laboratory monitoring from 2001 to 2008. Only new users of spironolactone co-prescribed with ARBs increased from 2001 to 2008 (P<0.001). In the 6 months before starting the co-prescriptions 96 to 100% of patients measured serum creatinine (mean 99.3%), sodium (97.3%) and potassium (98.6%). Within 3 months of starting the co-prescriptions 96 to 99% of patients measured serum sodium (mean 97.3%) and potassium (98.6%), but on average only 48% of them (range 43 to 53%) measured serum creatinine. Our results support the need for greater awareness within the medical community of the potential renal toxicity of the association of spironolactone with ACE-Is and/or ARBs. Adequate short-term monitoring of serum creatinine in all older community-dwelling people who receive such co-prescription is necessary in order to ensure safe usage of these medications.

‐ Electrocardiographic monitoring for new prescriptions of quetiapine co-prescribed with acetylcholinesterase inhibitors or memantine from 2005 to 2009. A population study on community-dwelling older people in Italy. Overall 2,623 community-dwelling older people started therapy with quetiapine, co-prescribed with acetylcholinesterase inhibitors AChEIs or memantine from 2005 to 2009. At least one electrocardiographic ECG was performed in 714 cases (27%) in the 6 months before-and in 398 cases (15%) within 3 months after-the starting of this prescription. ECG monitoring was performed both before and after starting quetiapine in only 160 cases (6%). At multivariable analyses, number of drugs taken, beta-blocker and antiarrhythmic drug use were found to be independent correlates of ECG monitoring whereas female sex was associated with a lower probability of receiving an ECG within 3 months after the initiation of quetiapine (odds ratio 0.78, 95 % CI 0.62-0.98). In conclusion, this study showed that ECG monitoring for new prescriptions of quetiapine in older people suffering from behavioural and psychological symptoms in dementia was actually performed infrequently, independently of the age of drug users, especially in women. Our results support the need for greater awareness within the medical community of the importance of such ECG monitoring.

B. DRUG UTILIZATION STUDIES

‐ Within region differences in outpatient antibiotic prescription. A total of 3 120 851 people (34 % of the population) received at least one antibiotic drug prescription. The highest prescription prevalence was observed in the 0-17 and 80 or more year age ranges (41.6% and 41.9%, respectively). Large differences were found in prevalence rates between different LHUs (ranging from 28.7% in Milan to 39.4% in Brescia) and in DIDs (ranging from 12.2 DID in Sondrio to 19.8 DID in Brescia). The age and residence of the population were the main determinants of drug exposure. In particular, patients aged <18 years (OR= 1.73; 95% CI 1.73, 1.74), aged 65 or older (OR= 1.64; 95% CI 1.63, 1.65), and those that live in Brescia (OR 1.66, 95% CI 1.65, 1.66) had a statistically significant higher risk of antibiotic drug exposure.

‐ Antipsychotic use in a sample of Italian Alzheimer Special Care Units. In institutionalised patients with dementia in northern Italian Alzheimer’s special care units (ASCU), 60% of 319 patients were taking at least one antipsychotic, 49% typical and 51% atypical. Forty five percent were exposed to one antipsychotic, 14% two and 1% three. Risperidone was the most frequently prescribed antipsychotic followed by promazine, olanzapine and haloperidol. In 40% of the cases, another hypnotic or sedative drug was simultaneously administered. Antipsychotics were significantly associated with female sex, older age and higher NPI score, but did not significantly influence mortality, hospitalisation, falls or use of physical restraint at follow-up.

‐ Comparison of Health Care Resource Utilization by Immigrants Versus Native Elderly People. For each immigrant (an older people born out of Italy), one person born in Lombardy (native) was randomly selected and matched by age, sex and general practitioner. The 25,508 immigrants selected were less prescribed with at least one drug (OR 0.72, 95 % CI 0.67-0.76) and had a lesser use of health care services (OR 0.79, 95 % CI 0.75-0.84) than natives. No statistically significant differences were found for hospital admission rates (OR 0.99, 95 % CI 0.99-1.04). A lower rate of health care resource

utilization was observed in elderly immigrants who had been living in the host region for as many as 10 years.

c. EPIFARM-ELDERY PROJECT

‐ Drug utilisation in elderly patients. All prescription for elderly aged 65 years or older (n=1 767 239), reimbursed by the National Health Service (NHS) in the Lombardy Region between 1 January and 31 December 2005 were analyzed. During the year of the study, 1555142 elderly (88% of the elderly population) received at least one drug prescription (89% women and 87% men). The overall prescription prevalence rate was slightly higher in women than in men (OR 1.20; 95% CI: 1.19-1.21), and increased up to 75 years of age in both sexes, reaching a plateau which persisted until 85 years. Each treated elderly received an average of 5 drugs (active substances) (median 4, interquartile range 2-7), without any difference between genders. Age and LHU of residence were predictors for chronic polypharmacy exposure and at multivariate analysis, elderly in age groups of 75-79, 80-84 and 85-89 years had the highest risk to be exposed to chronic polypharmacy (OR 2.25; 95%CI: 2.23-2.27, OR 2.68; 95%CI: 2.65-2.71, and OR 2.84; 95%CI: 2.79-2.89 respectively).

‐ Pattern of Cholinesterase Inhibitors Use in Alzheimer’s disease: Results of the EPIFARM-Elderly Project. The rate of elderly who received at least one prescription of cholinesterase inhibitors (ChEIs) increased from 0.5% in 2002 to 0.7% in 2004 and then remained unchanged until to 2007. The percentage of mild to moderate Alzheimer’s disease (AD) cases taking ChEIs was rather low (19-20%), and fairly stable overtime in the less treated oldest age groups (80+), while decreased in the youngest (65-79 years). In incident AD cases, the percentage of newly treated patients decreased overtime in the overall group (from 11.7% in 2004 to 8.0% in 2007) as well as in each age class. In the cohort of incident AD cases who started the treatment during 2004, nearly 40% were also in treatment three years later.

‐ Co-prescription of antipsychotics in patients treated with cholinesterase inhibitors (ChEIs): the EPIFARM-Elderly Project. Co-prescribing of atypical antipsychotics in patients exposed to ChEIs declined from 21.0% in 2002 to 14.6% in 2008 (OR 0.92; 95%CI:0.90, 0.94; p<0.001), while the prescribing prevalence of typicals slightly increased (OR 1.08; 95%CI:1.03, 1.13; p=0.001). In relation to the two warnings, the prevalence of patients who received a co-prescription of antipsychotics was significantly lower in 2005 than 2004 (23.1% vs. 28.0%; OR 0.79; 95%CI:0.73-0.86; p<0.001) and in 2007 than 2006 (19.4% vs. 23.0%; OR 0.79; 95%CI:0.73-0.86; p<0.001). After the first safety warning the prevalence of prescriptions for risperidone and olanzapine dropped significantly, and there was a significant increase for quetiapine. Haloperidol prescriptions increased, especially after the second warning. Despite regulatory warnings issued to discourage the use of antipsychotics, they are still frequently prescribed to patients taking ChEIs.

‐ Geographical differences in the prevalence of chronic polypharmacy in elderly people. This study analyzed nearly two million patients aged 65-94 years recorded in the Drug Administrative Database of the Lombardy Region (Northern Italy) from 2000 to 2010. Chronic polypharmacy was defined as taking five or more drugs in one month for at least six months (consecutive or not) in a year. Our results showed clusters of high and low prevalence rates of chronic polypharmacy and they were not influenced by age. Chronic polypharmacy was just weakly correlated with hospital admission (2000: ρ=0.08, p=0.0032; 2005: ρ=0.11, p<0.0001; 2010: ρ=0.18, p<0.0001), but not with mortality.

‐ One-year outcome changes of community-dwelling elderly people exposed to chronic polypharmacy: a comparison between data from 2001 and 2009 of the EPIFARM-Elderly Project. Among community-dwelling elderly people, the prevalence of those with incident chronic polypharmacy was 22,822/1,567,575 (1.46%) in 2001, and 51,471/1,800,257 (2.86%) in 2009. Both among elderly with chronic polypharmacy and among controls, the outcomes occurred with a lower frequency in 2009 than in 2001. In univariable regression analyses, chronic polypharmacy was associated with a higher risk of hospitalization (HR 3.21, 95% CI 3.14-3.29), of institutionalization (OR 1.88, 95% CI 1.76-2.01) and of death (HR 2.27, 95% CI 2.24-2.30). Chronic polypharmacy remained an independent predictor of adverse outcomes also after adjusting for index year, sex, age class, and number of drugs. In univariable and multivariable analyses index year, sex, age class, and number of drugs were also statistically

significantly associated with the outcomes. In particular, the year 2001, an older age and a higher number of drugs were associated with a higher risk of events; men were more frequently hospitalized or more frequently died, but were less frequently institutionalized, compared with women.

‐ Angiotensin-converting enzyme inhibitors and motor neuron disease: an unconfirmed association. To investigate the association between angiotensin converting enzyme inhibitors (ACEIs) and motor neuron disease (MND), including amyotrophic lateral sclerosis (ALS), we performed a population-based nested case-control study. Data were obtained from a population registry and the administrative database of the Lombardy Region (Northern Italy) from 2000 through 2010. Included were 1,200 patients with newly diagnosed MND/ALS and 120,000 controls, randomly selected from the same population and matched for gender, age and area of residence. Exposure to ACEIs or ARBs was quantified using defined daily doses (DDDs). Cumulative DDD (cDDD), which indicates the duration of drug use, was estimated as the sum of dispensed DDDs in the preceding 5 years, excluding 1 year before the MND/ALS diagnosis. Overall exposure, levels of exposure, and individual drug use were all assessed. Overall, there was no significant association between MND/ALS and antecedent use of ACEIs or ARBs. Data were confirmed in multivariable models and in subgroups. In conclusions a protective role of ACEIs in MND was not confirmed; ARBs were also uneventful. Differences with the Taiwanese (that showed an inverse association between ACEIs and ALS) study can be explained by the populations at risk, that may differ in genetic background and susceptibility to the action of environmental exposures, including drugs.

‐ Emergency department visits in older people: pattern of use, contributing factors, geographical differences and outcomes. In order to assess pattern the of use of Emergency Departments (EDs), factors contributing to the visits, geographical distribution and outcomes in people aged 65 years or more living in the Italian Lombardy Region in 2012. About 2 million of older people were included in the analyses: 78% had no ED visit, 15% only 1, 7% 2 or more. Male sex, ≥85 years, high number of drugs, ED visits and hospital admissions in the previous year and the location of an ED within 10 km from the patient’s place were all factors associated with a higher risk to have more ED visits. Clusters of high and low prevalence of visits were found for occasional users. Most of patients (74%) with at least 1 visit were discharged to home: 83% with low and 51% with high emergency triage code at admission. In conclusions in older people several variables were associated to an increased risk to have a high number of ED visits. Most of the visits were done for non-urgent problems and significant geographic differences were observed for occasional users.

d. STUDY ON DRUG-DRUG INTERACTIONS IN ELDERLY POPULATION

‐ Drug interactions in elderly patients. Elderly who received at least two co-administered prescriptions were selected to assess the presence of DDIs. 9115 elderly (16%) were exposed to potentially severe drug-drug interactions and 61% were women. A total of 13.520 severe drug interactions were recognized, mainly involving cardiovascular drugs (56.8% of the cases). The prevalence of potentially severe DDI increased at rising of the patient’s age and of the number of chronic drugs prescribed. At univariate and multivariate analysis age and number of chronic drugs were associated with an increasing risk of DDIs. Elderly constitute a population at high risk of DDIs.

‐ Changes in co-prescribing warfarin and potentially interacting drugs and risk of major bleeding in community-dwelling elderly people. To analyze the rate and trend of co-prescribing warfarin and potentially interacting drugs (PIDs) and the risk of hospitalization for major bleeding in community-dwelling elderly people, a cohort of community-dwelling elderly people (aged 65 years or more) who received at least one prescription for warfarin during the period 2001-2007 was drawn from Lombardy Region administrative database (northern Italy) was analysed. Age, local health unit (LHU) of residence, number of drugs and co-prescribed PIDs were predictors of hospitalization for hemorrhage, but the risk decreased during the study period (OR 0.94; 95% CI, 0.89-0.99). Compared with prescribing warfarin alone, coprescribing antibacterial drugs, calcium antagonists, allopurinol, omeprazole and ranitidine increased the risk of hospitalization for major bleeds. Over time, the rate of users warfarin of alone increased, and the percentage of those co-prescribed of PIDs fell slightly (χ2 trend: 3.74; p<0.001). No differences were found in the interaction between the co-prescription of warfarin with PID and years of prescription.

‐ Adverse drug reactions caused by drug-drug interactions in elderly outpatients. This prospective cohort study was aimed to determine the incidence and characteristics of DDI-related ADRs among elderly outpatients (aged ≥60 years) as well as the factors associated with these reactions. The incidence of DDI-related ADRs was 6 %. Warfarin was the most commonly involved drug (37 % cases), followed by acetylsalicylic acid (17 %), digoxin (17 %), and spironolactone (17 %). Gastrointestinal bleeding occurred in 37 % of the DDI-related ADR cases, followed by hyperkalemia (17 %) and myopathy (13 %). The multiple logistic regression showed that age ≥80 years, a Charlson comorbidity index ≥4, consumption of five or more drugs, and the use of warfarin were associated with the occurrence of DDI-related ADRs. With regard to severity, approximately 37 % of the DDI-related ADRs detected in our cohort necessitated hospital admission. All DDI-related ADRs could have been avoided (87 % were ameliorable and 13 % were preventable). The incidence of ADRs not related to DDIs was 10 % (n�=�44). The incidence of DDI-related ADRs in elderly outpatients is high; most events presented important clinical consequences and were preventable or ameliorable.

e. STUDY ON EPILEPSY

‐ Validation of healthcare administrative data for the diagnosis of epilepsy. The best and most conservative algorithm included EEG and selected treatment schedules and identified 61/71 patients with epilepsy (sensitivity 85.9%, CI 76.0% to 92.2%) and 15 623/15 657 patients without epilepsy (specificity 99.8%,CI 99.7% to 99.8%). The positive and negative predictive values were 64.2% and 99.9%. Sensitivity (86.7%) and the positive predictive value (68.4%) increased only slightly when patients with single seizures were included. A diagnostic algorithm including EEG and selected treatment schedules is only moderately sensitive for the detection of epilepsy and seizures. These findings apply only to the Northern Italian scenario.

‐ Prevalence and incidence of epilepsy in a well-defined population of Northern Italy. The claim records from the administrative district of Lecco, Northern Italy (population 320,609), collected during the years 2000 through 2008, were the data source. Epilepsy was defined as two or more unprovoked epileptic seizures 24+ hours apart with neurological confirmation. The most valid and conservative algorithm to detect patients with epilepsy was the combination of EEG and AEDs. Using this algorithm, a 9-year period prevalence (2000-2008) of epilepsy was calculated. Incident epilepsy cases were identified as a subset of prevalent cases among patients not traced in the years 2000 through 2003. Annual incidence rates were calculated for the period 2004 through 2008. 2663 patients met the inclusion criteria, giving an overall period prevalence of 8.28 per 1,000 (women: 7.83; men 8.75). Prevalence ratios tended to slightly increase with age. 864 incident cases were found. The overall annual incidence rate was 52.82 per 100,000 (women 50.53; men 55.20). Incidence rates peaked in the oldest age group. After accounting for false positives and false negatives, the adjusted prevalence ratio was 5.95 and the adjusted incidence rate 46.68). Adjusted ratios and rates were still higher in men than in women, especially in the older age classes. The standardized prevalence ratio was 6.20 per 1000 and the standardized incidence rate was 48.35 per 100,000. In conclusion, the prevalence of epilepsy in the district of Lecco is comparable to other studies, while the incidence among the highest.

‐ Long-term prognosis of epilepsy, prognostic patterns and drug resistance: a population-based study. Seizures in most people with epilepsy remit but prognostic markers are poorly understood. There is also little information on the long-term outcome of people who fail to achieve seizure control despite the use of two antiepileptic drugs (drug resistance). We identified people with a validated diagnosis of epilepsy, in whom two antiepileptic drugs had failed, from primary care records. All were registered with one of 123 family physicians in an area of Northern Italy. We determined remission (uninterrupted seizure freedom lasting 2 years or longer) and prognostic patterns (early remission, late remission, remission followed by relapse, no remission).We found that 747 individuals (381 men) were followed for 11,045.5 person-years. 428 (59%) were seizure-free. The probability of achieving 2-year remission was 18% at treatment start, 34% at two years, 45% at five, 52% at ten and 67% at 20 years (terminal remission, 60%). Epilepsy syndrome and drug resistance were the only independent predictors of 2- and 5-year remission. Early remission was seen in 101 people (19%), late remission in 175 (33%), remission followed by relapse in 85 (16%), and no remission in 166 (32%). Treatment response was the only variable associated with differing prognostic patterns. In conclusion, the long-term prognosis of epilepsy

is favorable in most cases. Early seizure remission is not invariably followed by terminal remission and seizure outcome varies according to well-defined patterns. Prolonged seizure remission and prognostic patterns can be predicted by broad syndromic categories and the failure of two antiepileptic drugs

f. NETWORK ANALYSIS

‐ The Drug Prescription Network: a system-level view of drug co-prescription in community-dwelling elderly people. Networks are well suited to display and analyze complex systems that consist of numerous and interlinked elements. This study was aimed i) to generate a series of drug prescription networks (DPNs) displaying co-prescription in community-dwelling elderly people; ii) to analyse DPN structure and organization; iii) to compare various DPNs in order to unveil possible differences in drug co-prescription patterns across time and space. Data were extracted from the administrative prescription database of the Lombardy Region, Northern Italy, in 2000 and 2010. DPNs were generated, in which each node represents a drug chemical subclass, while each edge linking two nodes represents the co-prescription of the corresponding drugs to the same patient. At a global level, the DPN was a very dense and highly clustered network, while at the local level it was organized into anatomically homogeneous modules. In addition, the DPN was assortative by class, as similar nodes (representing drugs with the same anatomic, therapeutic and pharmacologic annotation) connected to each other more frequently than expected, which indicates that similar drugs are often co-prescribed. Finally, temporal changes in the co-prescription of specific drug subgroups (for instance, proton pump inhibitors) translated into topological changes of the DPN and its modules. Complementing more traditional pharmacoepidemiology methods, the DPN-based method allows appreciating (and representing) general trends in the co-prescription of a specific drug (e.g., its emergence as a heavily co-prescribed hub) in comparison with other drugs.

Study for the improvement of the appropriateness of drug prescription

‐ Rationalization of drug prescribing in patients resident in the Bergamo Local Health Authority. In a study aimed to improve the quality of drug prescribing of general practitioners (GPs) in selected therapeutic areas (non-steroidal anti-inflammatory drugs, proton pump inhibitors, antibiotics, and antihypertensive agents, conducted among 160 GPs of the Bergamo Local Health Authority, we found a reduction of inappropriate prescribing of nearly 3% in all the indicators of drug utilization and cost analyzed.

‐ Medication non-adherence among elderly patients newly discharged and receiving polypharmacy. Among 100 patients recruited information on medication adherence was available for respectively 89 and 79 patients at first and second follow-up. Non-adherence was reported for 49 patients (55.1%) at the first follow-up and for 55 (69.6%) three months from discharge. Voluntary withdrawal of a drug and change of dosage without medical consultation were the main reasons for non-adherence at both follow-ups. The number of drugs prescribed at discharge was related to medication non-adherence at both follow-up interviews. No association was found between age and non-adherence. Only 25 patients (28.1%) at the first follow-up and 20 (25.3%) at the second understood the reasons for their medications. Low medication adherence is a real, complex problem for older patients receiving polypharmacy. We found that the increasing number of drugs prescribed at hospital discharge is correlated to non-adherence and a high percentage of patients did not understand the purpose of their medications.

‐ Proton pump inhibitors and risk of hypomagnesemia

PPI-associated hypomagnesemia is a rare but potentially life-threatening side-effect that has recently emerged as a consequence of their massive use, but the prevalence has not been established. The aims of this observational study were: (i) to assess the prevalence of hypomagnesemia in a cohort of patients in an internal Medicine ward; (ii) to estimate the risk of hypomagnesemia related to PPI treatment and its duration and the concomitant use of diuretics. All patients admitted in the internal medicine ward of the Ospedale di Circolo in Busto Arsizio between February and November 2014 were enrolled. Among the 604 patients recruited hypomagnesemia was detected in 85 (14.1%) and 63 (74.1%) used PPI. The prevalence of hypomagnesemia was significantly higher among patients taking PPI (21.1% vs 7.2%, p<0.001). PPI was independently associated with higher risk of hypomagnesemia than in patients not receiving PPI, also after adjusting the analysis for known risk factors. Unlike other studies we found that PPI risk factors for hypomagnesemia were higher among patients taking PPI for less than one year,

although the severe hypomagnesemia was recorded among patients taking them for longer. We suggest physicians to carefully assess the appropriateness of PPI therapy before starting it, watching out for symptoms of hypomagnesemia even in short-term treatments and reconsider PPI therapy in patients with low serum magnesium levels

‐ Role of community pharmacies for the detection of potentially inappropriate xanthine oxidase inhibitor prescriptions Xanthine oxidase (XO) inhibitors are largely the treatment of choice for gout, but allopurinol is often inappropriately used for asymptomatic hyperuricemia. To investigate the appropriateness of XO inhibitor use in relation to evidence-based indications and examine the role of community pharmacies in the detection of signal of inappropriate prescriptions of these drugs we conducted an observational study in 8 community pharmacies. The study sample included 74 patients receiving XO inhibitors. We found that one third of patients reported to be treated for asymptomatic hyperuricemia and has never had a gout attack. Four asymptomatic hyperuricemic patients referred to receive allopurinol to treat hypertension. Inappropriate use of XO inhibitors was principally related to treatment of hyperuricemia in patients with cardiovascular diseases, although there is little evidence that allopurinol is useful in preventing of cardiovascular diseases. Community pharmacies may also have a central role in the pharmacovigilance activity, by contributing to the prevention and identification of potential inappropriate drug prescriptions.

‐ Role of community pharmacists in the detection of potentially inappropriate benzodiazepines prescriptions for insomnia 

‐ Regular and prolonged use of hypnotics should be avoided because of the risk of tolerance to effects, dependence and an increased risk of adverse events. In 2012, updated Beers criteria for potentially inappropriate medication in older adults suggested to avoid all benzodiazepines in older adults to treat insomnia. To investigate the appropriateness of benzodiazepines prescription for insomnia and explore the role that community pharmacists can have in identifying signals of potential inappropriate drug prescriptions we conducted an observational study in 8 community pharmacies. A total of 181 participants were interviewed. About half of respondents (n=81) reported to be treated for insomnia. Fifty-two patients (64%) were on long term treatment (> 3 years) while for thirteen patients (16%) duration of treatment was comprised between 1 and 3 years. Thirty-three patients were in favour of benzodiazepine-discontinuation but in all cases discontinuation was unsuccessful for incorrect modality of drug withdrawals. Among patients treated for insomnia, seven received more than one benzodiazepine: two benzodiazepines in four cases, three in 2 cases and four in one case. Use of community pharmacy survey data allowed us to obtain information about incorrect management of insomnia and inappropriate benzodiazepines prescriptions.

‐ The ELICADHE-AIFA Project.

Effect of an integrated e-learning intervention, focused on “Comprehensive Geriatric Assessment” to improve the quality of drug prescribing in hospitalized elderly patients. With the aim to evaluate whether an integrated e-learning program of medical education, focused on teaching and implementing Comprehensive Geriatric Assessment (CGA) added to geriatric pharmacological notions (GPNs) (intervention) is superior to delivering only GPNs (control) in reducing the prescription of potentially inappropriate drugs (PID) or potential drug-drug interactions (PDDI) in hospitalized elderly, a cluster randomized single-blind controlled study was set up in a sample of elderly patients (aged 75 years or more) consecutively admitted to 20 geriatric and internal medicine hospital wards, and randomized to study intervention or control group. Secondary aims are to assess the clinical impact of the integrated e-learning intervention on the length of hospitalization, in-hospital and overall mortality, re-hospitalization, institutionalization and persistence of the effect of improving quality of drug prescribing during a follow-up of 12 months. A total of 697 patients (347 in the intervention and 350 in the control arms) were enrolled. No statistically significant difference in the prevalence of PIM at discharge was found between arms (OR 1.29 95%CI 0.87-1.91). We also found no decrease in the prevalence of DDI (OR 0.67 95%CI 0.34-1.28) and potentially severe DDI (OR 0.86 95%CI 0.63-1.15) at discharge, nor in mortality rates and incidence of re-hospitalization at 12 month follow-up. This e-learning educational program had no clear effect on the quality of drug prescription and clinical outcomes in hospitalized elderly patients. Given the high prevalence of PIMs and potential DDIs recorded in the frame of this

study, other approaches should be developed in order to improve the quality of drug prescription in this population

‐ FARMAGOOD-BIOSIMILARS PROJECT

The FARMAGOOD-BIOSIMILARS Project is a collaborative study between the IRCCS-Istituto di Ricerche Farmacologiche Mario Negri (IRFMN) and the Health Directorate of the Lombardy Region aimed to improve the appropriate use of biosimilars through several interventions shared with physicians. These intervention will be aimed:

‐ to promote the appropriateness of care pathways and rationalize the requirements of biological medicinal products "originator" and "biosimilars";

‐ monitor the benefit-risk profile of the use of these drugs in clinical practice (real life utilization); ‐ to save and free up resources in the pharmaceutical and healthcare spending.

The project is carried out in collaboration between the Foundation IRCCS Cà Granda Ospedale Maggiore Policlinico of Milan (where the Service SM-VAP will be realized) and will be divided into two phases: 1. Organization of a multidisciplinary team of internists, geriatricians, hospital pharmacists, nurses and

general practitioners (GPs), who will constitute the operational staff of the service. Planning and standardization of procedures of the evaluation process in accordance with the methodologies of Evidence Based Medicine.

2. Feasibility study and assessment of the impact of the service. The feasibility study will involve a sample of five hospital wards (internal medicine and geriatrics) and 500 patients aged > 65 years, with multimorbidity (at least two chronic conditions) and in polytherapy (treated with 5 or more drugs). The evaluation service will proceed to the review of therapeutic profiles and send to the clinicians a report with suggestions of how to improve the appropriateness of drug prescription and how to manage in each patient the drug-related problems emerged during the evaluation. If these counselling will be shared and accepted by the clinician and the patient, the adjustments-variations in the therapeutic profile will be implemented. At 1 and 3 months there will be a follow-up to verify the degree of persistence of the new therapeutic regimen.

Study on multimorbidity and polypharmacy in hospitalized elderly patients

The REPOSI Registry

The REgistro POliterapie SImi (REPOSI) study is a collaborative effort between the Italian Society of Internal Medicine (SIMI-Società Italiani Medicina Interna) and the Mario Negri Institute for Pharmacological Research. It was designed with the purpose to set up a network of internal medicine and geriatric wards in order to investigate patients aged 65 years or older affected by multiple diseases and prescribed with polypharmacy. Participation to the network was on a voluntary basis. During a period of four weeks, three months apart each from the other, the 38 wards involved in the study, recruited 1332 elderly patients (aged 65 years or older). The main results from the analyses of this cohort of hospitalized elderly patients are the following:

1. at hospital admission 52% of patients taken five or more different drugs (polypharmacy) and were in the ward for a mean of 11 days.

2. The comparison discharge-admission showed an increasing rate of patient with polypahramacy (+13%) and with multiple disease (+16%).

3. No difference emerged in terms of in-hospital mortality between patients with polypharmacy and the other ones.

4. At multivariate analysis the in-hospital mortality and hospital stay were positively associated with age, adverse clinical events, and comorbidity (Charlson Index).

Furthermore, with aim of recognizing clusters of diseases among the hospitalized elderly, and of identifying groups of patients at risk of in-hospital death and adverse clinical events according to disease clustering, a

regression analysis was done. Patients affected by the clusters including heart failure (HF) and either chronic renal failure (CRF), or chronic obstructive pulmonary disease had a significant association with in-hospital death (OR=4.2;95%CI=1.6-11.4; OR=2.9;95%CI=1.1-8.1, respectively), as well as patients affected by CRF and anaemia (OR=6.0;95%CI=2.3-16.2). The cluster including HF and CRF was also associated with adverse clinical events (OR=3.5;95%CI=1.5-7.7). The effect of both HF and CRF and CRF and anaemia on in-hospital death was additive.

Other analyses:

‐ Among 1155 patients eligible for the analyses, elderly treated with drugs for the treatment of gastro-oesophageal reflux disease (GERD) or peptic ulcer were 466 (40.3%) at hospital admission and 647 (56.0%) at discharge. 65.2% of patients receiving a drug for peptic ulcer or GERD at admission and 64.1% at discharge were inappropriately treated. Among patients inappropriately treated the number of other drugs prescribed was associated with an increased use of drugs for peptic ulcer or GERD, also after adjustment for age, sex and number of diagnoses at admission (OR 95%CI=1.25 (1.18-1.34), p=.0001) or discharge (OR 95%CI= 1.11 (1.05-1.18), p=0.0003).

‐ A statistically significant association between the presence of bacterial infection and use of PPI was found. This association was greater in elderly receiving the drug for more than 14 days and even after adjusting results for age, sex and comorbidity.

‐ Among patients with AFF 26.5% at admission and 32.8% at discharge were not on antithrombotic therapy, and 43.7% at admission and 40.9% at discharge were not taking an appropriate therapy according to the CHADS2 score. Among elderly patients admitted with a diagnosis of AFF to internal medicine wards, an appropriate antithrombotic prophylaxis was taken by less than 50%, with an underuse of VKAs prescription independently of the level of cardio-embolic risk. Hospitalization did not improve the adherence to guidelines.

‐ After multiadjustment, the diagnosis of dementia was associated with in-hospital death (OR = 2.1; 95% CI = 1.0 - 4.5). Having dementia and at least one adverse clinical event during hospitalization showed an additive effect on in-hospital mortality (OR = 20.7 ;95% CI = 6.9 – 61.9). Acutely ill elderly patients affected by dementia are more likely to die shortly after hospital admission. Having dementia and adverse clinical events during hospital stay increases the risk of death.

‐ Among clusters of diseases, the highest mean number of drugs (N=8) was found in patients affected by heart failure (HF) plus chronic obstructive pulmonary disease (COPD), HF plus chronic renal failure (CRF), COPD plus coronary heart disease (CHD), diabetes mellitus plus CRF, and diabetesmellitus plus CHD plus cerebrovascular disease (CVD). The strongest association between clusters of diseases and polypharmacy was found for diabetes mellitus plus CHD plus CVD, diabetes plus CHD, and HF plus atrial fibrillation (AF).

‐ The number of patients treated with antidepressant medication at hospital admission was 115 (9.9%) and at discharge 119 (10.3%). In a multivariate analysis, a higher number of drugs (OR = 1.2; 95% CI = 1.1–1.3), use of anxiolytic drugs (OR = 2.1; 95% CI = 1.2–3.6 and OR = 3.8; 95% CI = 2.1–6.8), and a diagnosis of dementia (OR = 6.1; 95% CI = 3.1–11.8 and OR = 5.8; 95% CI = 3.3–10.3, respectively, at admission and discharge) were independently associated with antidepressant prescription. A specific diagnosis requiring the use of antidepressants was present only in 66 (57.4%) patients at admission and 76 (66.1%) at discharge.

‐ 19% of patients were re-admitted at least once within 3 month after discharge. Multivariate logistic regression analysis showed that only AEs during hospitalization, previous hospital admission, and vascular and liver diseases were significantly associated with likelihood of readmission.

‐ There was a dose-response relation between total Anticholinergic Cognitive Burden (ACB) score and cognitive impairment. Patients identified by the Anticholinergic Risk Scale (ARS) had more severe cognitive and physical impairment, and the dose-response relation was clear for ability in activities of daily living. No correlation was found with length of hospital stay.

‐ Multivariate analysis found a significant association with an increased risk of mortality at 3 months follow in patients exposed to at least 2 potentially severe DDIs (OR=2.62; 95% CI, 1.00-6.68; p=0.05). The cause of adverse clinical events was potentially related to severe DDIs in 2 patients who died during

hospitalization, in 5 patients re-hospitalized and in one who died at 3 months follow-up after discharge. Hospitalization was associated to an increase in potentially severe DDIs.

‐ Prevalence of patients receiving almost one PIM according to the Beers criteria were 20.1% and 23.5% with the 2003 and 2012 version, respectively. Prescription of PIMs according to those criteria was not associated with an increased risk of adverse clinical events, re-hospitalization and all-cause mortality at three month follow-up in both univariate and multivariate analysis, after adjusting for age, sex and CIRS comorbidity index. On the other hand, anticholinergic drugs assessed according to the ACB scale was associated with an increased risk of re-hospitalization in both univariate (OR=1.73, 95%CI, 1.16-2.56, p=0.006) and multivariate models (OR=1.67 95%CI, 1.12-2.53, p=0.01).

‐ In univariate and multivariable analyses, a higher bleeding score was negatively associated with warfarin prescription, and positively associated with aspirin prescription. The cardio-embolic scores were associated with the therapeutic choice only after adjusting for bleeding score or age.

‐ Multivariable logistic regression analysis showed that severely reduced eGFR at the time of admission was associated with in-hospital mortality (OR 3.00; 95 % CI 1.20-7.39, p = 0.0230), but not with re-hospitalization (OR 0.97; 95 % CI 0.54-1.76, p = 0.9156) or mortality at 3 months after discharge (OR 1.93; 95 % CI 0.92-4.04, p = 0.1582). On the contrary, an increased risk (OR 2.60; 95 % CI 1.13-5.98, p = 0.0813) to die within 3 months after discharge was associated with decreased eGFR measured at the time of discharge.

‐ Among 2.712 patients eligible for the analysis, 303 (11.2%) were treated with allopurinol at hospital admission and 292 (12.6%) among 2.314 patients discharged. Only 16 (5.3%) of patients receiving allopurinol at admission and 22 (7.5%) at discharge were appropriately treated. Among these, asymptomatic hyperuricemia, polytherapy, chronic renal failure, diabetes, ischemic cardiomyopathy, BPCO and atrial fibrillation was significantly associated with greater use of allopurinol. Prevalence of inappropriate prescription of allopurinol remained almost the same at admission and discharge.

‐ Only 38.8% of patients with a diagnosis of pneumonia received an empirical antibiotic regimen adherent to guidelines. However, no significant association was found between adherence to guidelines and outcomes. Having HAP, older age and higher CIRS Severity Index were the main factors associated with in-hospital mortality.

‐ The main gender differences in-patients included in the REPOSI registry were: polypharmacy (> 5 drugs) was more frequent in men both at hospital admission and discharge. Neuropsychiatric drugs were significantly more prescribed in women (p<0.0001); at admission men were more likely to be on antiplatelets (41.7% vs 36.7%; p=0.0029), ACE-inhibitors (28.7% vs 24.7%; p=0.0072 ) and statins (22.9% vs 18.3%; p=0.0008). At discharge, antiplatelets (43.7% vs 37.3%; p=0.0003) and statins (25,2% vs 19.6%; p<0.0001) continued to be prescribed more often in men, while women were given beta blockers more often than men (21.8% vs 18.9%; p=0.0340). Proton pump inhibitors were the most prescribed drugs regardless of gender.

‐ Women were older than men, more often widow and living alone or in nursing homes. Disease distribution showed that malignancy, diabetes, coronary artery disease, chronic kidney disease and chronic obstructive pulmonary disease were more frequent in men, but hypertension, osteoarthritis, anemia and depression were more frequent in women. Severity and comorbidity indexes according to the Cumulative Illness Rating Scale (CIRS-s and CIRS-c) were higher in men, while cognitive impairment evaluated by the Short Blessed Test (SBT), mood disorders by the Geriatric Depression Scale (GDS) and disability in daily life measured by Barthel Index (BI) were worse in women. In-hospital and 3-month mortality rates were higher in men.

‐ To investigate the social and clinical profiles of frailty and to study their association with related outcomes in hospitalized elderly patients (65 years or more) we performed a Multiple Correspondence Analysis (MCA) on data collected in 2010 and 2012. MCA identified two principal latent variables that may explain the “burden of diseases” and the “cognitive and functional impairment” of patients. The presence of three clusters were also pointed out: the first identified “healthy” subjects, the second distinguished patients with important organ diseases and the third identified the oldest old patients with cognitive and physically functional impairments. Significant higher in-hospital mortality was found among patients in cluster II and Cluster III (2.6% and 4.8% vs 0.6%) than among those in Cluster I. Also

significant higher re-hospitalization rate was observed among patients in Cluster II (19.8%). In conclusion this work enables us to describe frailty as a progressive manifold decline, and in particular it allows to distinguish frail elderly patients with organ diseases and functional reserve loss, and patients with cognitive impairment and psychological disorders. Our results also confirm that both frailty phenotypes are predictive of unfavorable outcomes.

‐ To evaluate the prevalence of QT-prolonging drugs prescription at hospital admission and discharge and risk factors associated with their use in older people, we analyzed data obtained from 4035 patients enrolled in 2008 (n=1332), 2010 (n=1380) and 2012 (n=1323). Among 3906 patients prescribed with at least one drug at admission, 2156 (55.2%) were taking at least one QT-prolonging drug. Risk factors independently associated with the use of any QT-prolonging drugs were found to be increasing age (OR 1.02, 95%CI 1.01-1.03), multimorbidity (OR 2.69, 95%CI 2.33-3.10), hypokalemia (OR 2.79, 95%CI 1.32-5.89), atrial fibrillation (OR 1.66, 95%CI1.40-1.98), and heart failure (OR 3.17, 95%CI 2.49-4.05). Furosemide, alone or in combination, was the most prescribed drug. Amiodarone was the most prescribed drug with a definite risk of TdP. Both the absolute number of QT-prolonging drugs (2890 vs 3549) and the number of patients treated with them (2456 vs 2156) increased at discharge. Among 1808 patients not prescribed QT-prolonging drugs at admission, 35.8% were prescribed with them at discharge. In conclusion, despite their risk, QT-prolonging drugs are widely prescribed to hospitalized older persons in internal and geriatric wards.

Assessment of quality of services for eledelry people

‐ Quality assessment of services on dementia. A sample of Lombardy Region Alzheimer Special Care Units (ASCU) was compared with traditional nursing homes to assess their effects on main clinical outcome in a sample of 450 residents followed for 18 months. Patients admitted at ASCU had a lower risk of hospitalisation, use of physical restraints, and a higher probability of withdrawing antipsychotics than patients admitted to NH. No difference was reported on overall mortality and falls.

‐ Census and quality assessment of the Lombardy Region Alzheimer Evaluation Unit (AEU). A collaborative study with the Italian Alzheimer Association (Federazione Alzheimr Italia) was organised with the aim to assess the quality of Lombardy Region AEU. After a census of the 81 AEU active in the Lombardy Region, a random sample of 18 AEU was selected for the quality evaluation by specific indicators that covered all the three axes of quality (structure, process and outcome). The overall quantitative score for each of the three axes was nearly 50% of the available score. The comparison of the 18 AEU sowed some differences in all the three quality axes, in particular the process axis. The results of the study highlight the need to improve the standard of these services in order to better meet the needs of families and patients with Alzheimer Disease.

‐ Caregivers’ perceptions of the therapeutic benefits of cholinesterase inhibitors. The aim of the study was to collect opinions, perceptions, and expectations on the therapeutic benefits of ChEIs and the impact on the care of the patient in a large sample of caregivers. This used an ad-hoc online questionnaire that was accessible for nearly four months on the Federazione Alzheimer Italia website and had three sections: 1) information on the patient with dementia; 2) information on the caregiver’s perception of the therapeutic benefits of ChEIs; 3) information on caregivers. During the access time, 439 questionnaires were filled, and 369 were validated for inclusion in the analysis; of these, 329 also had information on caregivers. Caregivers’ beliefs about the effectiveness of dementia treatment, their expectations and changes in their lives were clear.

Computerized prescription support system (intercheck®)

(https://clinicalweb.marionegri.it/intercheckweb/)

‐ Computerized Prescription Support Systems (CPSSs) are programs or software developed to highlight inappropriate prescribing and minimize the occurrence of adverse drug reactions (ADRs). We developed INTERcheck® in order to optimize drug prescribing in elderly people with complex co-morbidity and altered pharmacokinetics and pharmacodynamics. To evaluate the effectiveness of INTERcheck to review pharmacological profile and reduce the use of potentially inappropriate medications (PIMs), potentially severe drug-drug interactions (DDIs) and the correlation between the clinical relevance of

pDDIs as detected by INTERCheck® and the clinician's personal judgement we conducted some study in different setting: geriatric ward, nursing homes and hospital pharmacy.

‐ Prevention of inappropriate prescribing in hospitalized older patients using a Computerized Prescription Support System (INTERcheck) We conducted a prospective study on two samples of elderly patients hospitalized in an Italian geriatric ward. In the observational phase the number of patients exposed to at least one PIMs remained unchanged from admission (n=29, 39.1%) to discharge (n=28, 37.8%). In the intervention phase 25 patients (41.7%) were exposed to at least one PIMs at hospital admission and 7 (11.6%) at discharge (p<0.001). Similarly patients exposed to at least one potentially severe DDI decreased respectively from 27 (45.0%) to 20 (33.3%), p=0.703. The number of newly created potentially severe DDIs decreased from 37 (59.0%) of the observational phase to 9 (33%.0), p<0.001. Use of INTERCheck was associated to a significant reduction in PIMs and potentially severe DDIs.

‐ A multicomponent intervention to optimize psychotropic drug prescription in elderly nursing home residents: a Italian multicentre, prospective, pilot study The aim of this multicenter prospective study was to optimize the prescription of psychotropic drugs in a sample of older patients living in nursing homes in Italy, through a multicomponent intervention; education of general practitioners and the use of INTERcheck. Three educational interventions (‘ex cathedra’ presentations) were organized by the researchers involved in the project and a fourth training session was also held on the use of INTERCheck. Among the 272 patients included in the analysis there was a significant reduction in the mean number of drugs and in the percentage receiving psychotropic drugs and potentially inappropriate psychotropic drugs (respectively 11.5% and 30.6%, p<0.0001). Patients exposed to at least one potentially severe DDI also decreased from 145 (53.3%) at the first time-point to 87 (32.0%) at the last (p<0.0001). Results were confirmed in the 181 patients for whom information about drug treatment were available at all time-points. The combination of an educational intervention and the CPSS can achieve a significant reduction in potentially inappropriate psychotropic drug use, psychotropic duplicates and potentially severe DDIs in nursing homes.

‐ The value of software that provides clinically relevant information on drug interactions In order to define the clinicians' personal judgement about the potential drug-drug interactions (pDDIs) detected by INTERCheck, the hospital pharmacy of the Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico of Milan prepared four questions for each pDDIs: 1) is the actual pDDI known? 2) is it clinically relevant? 3) if yes, why? 4) would knowledge of the pDDI at the time of prescription have changed patient management (in terms of prescribing approach or clinical monitoring)? The questionnaire was administered to clinicians of three internal medicine wards and one geriatric ward. The revision of 60 medical records using INTERCheck identified 448 pDDIs evaluated by the clinicians. Overall, 51% of pDDIs (230/448) were judged clinically relevant, with a rate increasing according to the severity declared by the CPSS. Of 230 pDDIs, 67% was considered relevant for clinicians due to their potential clinical impact, 24% owing to patient complexity and/or comorbidities, and 7% because they were unknown. More than half of the electronically retrieved pDDIs were considered clinically relevant by clinicians and in 23% of the cases, CPSS use would have led to a different patient management, such as drug withdrawal, dose adjustment or clinical and biochemical monitoring.

Drug information service for the elderly

‐ A daily free of charge telephone service for drug and clinical information is available for physicians and elderly. Nearly 600 questions are answered each year.