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University of Pavia

13 ARTICLES PUBLISHED IN JoVE

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Neuroscience

A Novel Strategy Combining Array-CGH, Whole-exome Sequencing and In Utero Electroporation in Rodents to Identify Causative Genes for Brain Malformations
Valerio Conti *1, Aurelie Carabalona *2,3,15, Emilie Pallesi-Pocachard 2,3,4, Richard J. Leventer 5,6,7, Fabienne Schaller 2,3,8, Elena Parrini 1, Agathe A. Deparis 2,3, Françoise Watrin 2,3, Emmanuelle Buhler 2,3,8, Francesca Novara 9, Stefano Lise 10, Alistair T. Pagnamenta 10, Usha Kini 11, Jenny C. Taylor 10, Orsetta Zuffardi 9,12, Alfonso Represa 2,3, David Antony Keays 13, Renzo Guerrini 1,14, Antonio Falace 2,3, Carlos Cardoso 2,3
1University of Florence, 2INSERM INMED, 3Aix-Marseille University, 4Plateforme Biologie Moléculaire et Cellulaire INMED, 5Royal Children's Hospital, 6Murdoch Children's Research Institute, 7University of Melbourne, 8Plateforme postgenomique INMED, 9University of Pavia, 10Wellcome Trust Centre for Human Genetics, 11Oxford Radcliffe NHS Trust, 12IRCCS Casimiro Mondino Foundation, 13Research Institute of Molecular Pathology, 14IRCCS Stella Maris, 15Columbia University

Periventricular nodular heterotopia (PNH) is the most common form of malformation of cortical development (MCD) in adulthood but its genetic basis remains unknown in most sporadic cases. We have recently developed a strategy to identify novel candidate genes for MCDs and to directly confirm their causative role in vivo.

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Medicine

Tissue Characterization after a New Disaggregation Method for Skin Micro-Grafts Generation
Valeria Purpura 1, Elena Bondioli 1, Antonio Graziano 2, Letizia Trovato 2, Davide Melandri 1, Martina Ghetti 1, Andrea Marchesini 3, Maria Gabriella Cusella De Angelis 4,5, Laura Benedetti 4,5, Gabriele Ceccarelli 4,5, Michele Riccio 3
1Burns Centre and Emilia Romagna Regional Skin Bank, 2Human Brain Wave srl, 3Plastic and Reconstructive Surgery, AOU “Ospedali Riuniti”, 4Department of Public Health, Experimental Medicine, Anatomy and Forensic, University of Pavia, 5C.H.T Centre for Health Technologies, University of Pavia

The protocol describes a new method to disaggregate human tissues and to create autologous micro-grafts that, combined with collagen sponges, give rise to human bio-complexes ready to use in the treatment of skin lesions. Further, this system preserves cell viability of micro-grafts at different times after mechanical disaggregation.

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Developmental Biology

Isolation and Characterization of Mouse Antral Oocytes Based on Nucleolar Chromatin Organization
Manuela Monti 1, Carlo Alberto Redi 1,2
1Research Center for Regenerative Medicine, Fondazione IRCCS Policlinico San Matteo, 2Department of Biology and Biotechnology "L. Spallanzani", University of Pavia

Here we present a protocol for the characterization of mouse antral oocytes based on nucleolar organization.

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Genetics

Delivery of Nucleic Acids through Embryo Microinjection in the Worldwide Agricultural Pest Insect, Ceratitis capitata
Paolo Gabrieli 1, Francesca Scolari 1
1Department of Biology and Biotechnology, University of Pavia

The Mediterranean fruit fly (medfly) Ceratitis capitata (Diptera: Tephritidae) is a worldwide pest of agriculture. A deeper understanding of its biology is key to control medfly populations and thus reduce economic impact. Embryo microinjection is a fundamental tool allowing both germ-line transformation and reverse genetics studies in this species.

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Bioengineering

An Experimental Protocol for Assessing the Performance of New Ultrasound Probes Based on CMUT Technology in Application to Brain Imaging
Giulia Matrone 1, Alessandro Ramalli 2, Alessandro Stuart Savoia 3, Fabio Quaglia 4, Gloria Castellazzi 5, Patrizia Morbini 6, Marco Piastra 1
1Department of Electrical, Computer and Biomedical Engineering, University of Pavia, 2Department of Information Engineering, University of Florence, 3Department of Engineering, Roma Tre University, 4FTMTR&D/SPA, STMicroelectronics, 5Brain Connectivity Center, BCC, Istituto Neurologico Nazionale Fondazione C. Mondino I.R.C.C.S., 6Department of Molecular Medicine - Unit of Pathology, University of Pavia, Foundation IRCCS Policlinico San Matteo

The development of new ultrasound (US) probes based on Capacitive Micromachined Ultrasonic Transducer (CMUT) technology requires an early realistic assessment of imaging capabilities. We describe a repeatable experimental protocol for US image acquisition and comparison with magnetic resonance images, using an ex vivo bovine brain as an imaging target.

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Developmental Biology

A Neural Network-Based Identification of Developmentally Competent or Incompetent Mouse Fully-Grown Oocytes
Federica Cavalera 1, Mario Zanoni 1, Valeria Merico 1, Thi Thu Hien Bui 1,2, Martina Belli 1,3, Lorenzo Fassina 4, Silvia Garagna 1, Maurizio Zuccotti 1
1Laboratorio di Biologia dello Sviluppo, Dipartimento di Biologia e Biotecnologie "Lazzaro Spallanzani", University of Pavia, 2Institute of Biotechnology, University of Helsinki, 3Department of Reproductive Medicine, University of California San Diego, 4Dipartimento di Ingegneria Industriale e dell'Informazione, University of Pavia

Here, we present a protocol for non-invasive assessment of oocyte developmental competence performed during their in vitro maturation from the germinal vesicle to the metaphase II stage. This method combines time-lapse imaging with particle image velocimetry (PIV) and neural network analyses.

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Immunology and Infection

Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers
Giulia Ercoli 1, Gianluca Lopez 1,2, Camilla Ciapponi 1,3, Chiara Corti 1,2, Luca Despini 4, Donatella Gambini 5, Letterio Runza 1, Concetta Blundo 4, Amedeo Sciarra 1, Nicola Fusco 1,6
1Division of Pathology, Fondazione IRCCS Ca' Granda, Ospedale Maggiore Policlinico, 2School of Medicine, University of Milan, 3School of Biology, University of Pavia, 4Division of Breast Surgery, Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico, 5Division of Medical Oncology, Fondazione IRCCS Ca' Granda - Ospedale Maggiore Policlinico, 6Department of Biomedical, Surgical, and Dental Sciences, University of Milan

Heterogeneous distribution of HER2-positive cells can be observed in a subset of breast cancers and generates clinical dilemmas. Here, we introduce a reliable and cost-effective protocol to define, quantify, and compare HER2 intra-tumor genetic heterogeneity in a large series of heterogeneously processed breast cancers.

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Cancer Research

Detection of a CDH1 Rare Transcript Variant in Fresh-frozen Gastric Cancer Tissues by Chip-based Digital PCR
Chiara Molinari 1, Raefa Abou Khouzam 2, Samanta Salvi 1, Tania Rossi 1, Guglielmina Nadia Ranzani 2, Daniele Calistri 1
1Biosciences Laboratory, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, 2Department of Biology and Biotechnology, University of Pavia

The manuscript describes a chip-based digital PCR assay to detect a rare CDH1 transcript variant (CDH1a) in fresh-frozen normal and tumor tissues obtained from patients with gastric cancer.

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Neuroscience

Meta-analysis of Voxel-Based Neuroimaging Studies using Seed-based d Mapping with Permutation of Subject Images (SDM-PSI)
Anton Albajes-Eizagirre 1,2, Aleix Solanes 1,2, Miquel Angel Fullana 2,3, John P. A. Ioannidis 4, Paolo Fusar-Poli 5,6,7, Carla Torrent 1,2,3,8, Brisa Solé 1,2,3,8, Caterina Mar Bonnín 1,2,3,8, Eduard Vieta 1,2,3,8, David Mataix-Cols 9, Joaquim Radua 1,2,5,9
1Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), 2Mental Health Research Networking Center (CIBERSAM), 3Institute of Neurosciences, Hospital Clinic de Barcelona, 4Departments of Medicine, of Health Research and Policy, and of Biomedical Data Science, Stanford University School of Medicine, and Department of Statistics, Stanford University School of Humanities and Sciences, 5Department of Psychosis Studies, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 6OASIS Service, South London and Maudsley NHS Foundation Trust, 7Department of Nervous System and Behavioral Sciences, University of Pavia, 8University of Barcelona, 9Centre for Psychiatric Research and Education, Department of Clinical Neuroscience, Karolinska Institutet

We detail how to conduct a meta-analysis of voxel-based neuroimaging studies using Seed-based d Mapping with Permutation of Subject Images (SDM-PSI).

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JoVE Core

Abbiategrasso Brain Bank Protocol for Collecting, Processing and Characterizing Aging Brains
Tino Emanuele Poloni *1,4, Valentina Medici *1, Arenn Faye Carlos 1, Annalisa Davin 2, Arcangelo Ceretti 1, Michela Mangieri 1, Paola Cassini 1,4, Roberta Vaccaro 3, Daniele Zaccaria 3, Simona Abbondanza 3, Matteo Bordoni 5, Valentina Fantini 2,6, Elena Fogato 7, Cristina Cereda 5, Mauro Ceroni 8, Antonio Guaita 1,2,3
1Department of Neurology and Neuropathology, Golgi-Cenci Foundation, 2Laboratory of Neurobiology and Neurogenetic, Golgi-Cenci Foundation, 3Department of Neuropsychology and Social Sciences, Golgi-Cenci Foundation, 4Department of Rehabilitation, ASP Golgi-Redaelli Geriatric Hospital, 5Genomic and Post-Genomic Center, IRCCS Mondino Foundation, 6Department of Brain and Behavioral Sciences, University of Pavia, 7Department of Pathology, ASP Golgi-Redaelli Geriatric Hospital, 8Department of Neurological Science, IRCCS Mondino Foundation, University of Pavia

This protocol describes a method to track individual brain-aging trajectories through a brain donation program and proper characterization of brains. Brain donors are involved in a long-term longitudinal study including serial multi-dimensional assessments. The protocol contains a detailed description of brain processing and an accurate diagnostic methodology.

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Behavior

Implementation of a Real-Time Psychosis Risk Detection and Alerting System Based on Electronic Health Records using CogStack
Tao Wang 1, Dominic Oliver 2, Yamiko Msosa 1, Craig Colling 3, Giulia Spada 2, Łukasz Roguski 4, Amos Folarin 1, Robert Stewart 3,5, Angus Roberts 1,3, Richard J. B. Dobson 1,3,4,6, Paolo Fusar-Poli 2,3,7,8
1Department of Biostatistics and Health Informatics, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 2Early Psychosis: Interventions and Clinical-detection (EPIC) lab, Department of Psychosis Studies, Institute of Psychiatry, Psychology & Neuroscience, King's College London, 3National Institute for Health Research, Maudsley Biomedical Research Centre, South London and Maudsley National Health Service (NHS) Foundation Trust, 4Institute of Health Informatics, University College London, 5Department of Psychological Medicine, Institute of Psychiatry, Psychology and Neuroscience, King's College London, 6Health Data Research UK London, University College London, 7OASIS service, South London and Maudsley National Health Service (NHS) Foundation Trust, 8Department of Brain and Behavioral Sciences, University of Pavia

We demonstrate how to deploy a real-time psychosis risk calculation and alerting system based on CogStack, an information retrieval and extraction platform for electronic health records.

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Bioengineering

Detergent-Free Decellularization of the Human Pancreas for Soluble Extracellular Matrix (ECM) Production
Riccardo Tamburrini 1,2,3, Deborah Chaimov 1,3, Amish Asthana 1,3, Carlo Gazia 3,4, Kevin Enck 3, Sean M. Muir 5, Justine Mariam Aziz 6, Sandrine Lablanche 7, Emily Tubbs 7, Alice A. Tomei 8,9, Mark Van Dyke 10, Shay Soker 3, Emmanuel C. Opara 3, Giuseppe Orlando 1,3
1Department of Surgery, Wake Forest Baptist Medical Center, 2Department of General Surgery, PhD Program in Experimental Medicine, University of Pavia, 3Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, 4Department of Surgery, Tor Vergata University of Rome, 5Wake Forest University College of Arts and Science, 6Wake Forest University School of Medicine, 7Laboratory of Fundamental and Applied Bioenergetics (LBFA), and Environmental and System Biology (BEeSy), Grenoble Alps University, 8Department of Biomedical Engineering, University of Miami, 9Diabetes Research Institute, University of Miami Miller School of Medicine, 10Department of Biomedical Engineering and Mechanics, School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University

The protocol described in this manuscript explains the steps for the fabrication of a soluble extracellular matrix (ECM) from the human pancreas. The solubilized ECM powder obtained through this protocol may be used for the recapitulation of pancreatic islets’ microenvironment in vitro and, potentially, in vivo settings.

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Biology

Isolation and Screening from Soil Biodiversity for Fungi Involved in the Degradation of Recalcitrant Materials
Marta Elisabetta Eleonora Temporiti 1, Chiara Daccò 1, Lidia Nicola 1
1Laboratory of Mycology, Department of Earth and Environmental Sciences, University of Pavia

Here, we present a protocol for screening soil biodiversity to look for fungal strains involved in the degradation of recalcitrant materials. First, fungal strains able to grow on humic acids or lignocellulose are isolated. Their activity is then tested both in enzymatic assays and on pollutants such as hydrocarbons and plastics.

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