Relazione 2007 - BTBS

‘07
A N N U A
R E P O R
L
T
DEPARTMENT OF
BIOTECHNOLOGY
AND BIOSCIENCES
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INDEX
INTRODUCTION
1
STRUCTURE AND ORGANIZATION
1.1
1.2
1.3
1.4
1.5
1.6
Department Management Structure and Staff
Organization and Structure
Instrumentation and Facilities
Teaching Activity
Advanced Training
Master and PhD theses
3
4
8
12
14
15
19
2
RESERCH GROUPS
25
3
SCIENTIFIC PUBLICATION INDEX, GRANTS
57
3.1
3.2
3.3
3.4
Research grants and contracts
Publications
Book chapters
Patents
58
60
66
67
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INTRODUCTION
This report is a description of the Department of Biotechnology and
Biosciences (BtBs) of the State University of Milano-Bicocca, its structure
and organization, its permanent, temporary and in training staff. It also includes information on financing and a detailed description of teaching and research activities and scientific results obtained.
The research areas of the Department benefit from multidisciplinary expertise in the fields of Biotechnology and Biosciences. Great attention is paid to
advanced topics that allow the Department to establish scientific cooperation
with other national and international bodies including industrial and pharmaceutical R&D departments.
About 230 people are active in a total usable floor space of 6.000 m2, working
on both basic and contract research as well as in teaching in the curricula of
Biotechnology, Biological Sciences and Bioinformatics. The Department’s
budget in 2007 was over 8 MIO euros coming from regional and national
sources, European research programs, research contracts and financing
from private foundations.
BTBS participates to different consortia such as BIOMILANO, BIOBRESSO
(Bioinformatics), CINMPIS (Innovative Methods and Processes in Chemical
Synthesis); GENOPOLIS (Functional Genomics), STEMGEN (Stem cells), IBC
(Biocatalysis), BARCODING and hosts a laboratory of the Foundation Rita Levi
Montalcini devoted to Neurosciences
FUNDING
STAFF
MIUR (PRIN, FISR, FIRB)
FULL PROFESSORS
EC GRANTS
ASSOCIATED PROFESSORS
REGIONE LOMBARDIA GRANTS
RESEARCHS
CARIPLO GRANTS
PhD STUDENTS
OTHER FUNDING AGENCIES
FELLOWS
REGIONE LOMBARDIA GRANTS
POST-DOCS
RESEARCH SERVICES
ADMINISTRATIVE STAFF
DEPARTMENT FUNDS
TECHNICAL STAFF
TEACHING RESOURCES
PhD COURSES
MASTER
OTHER RESOURCES
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1
S TRUCTURE AND
O R G A N I Z AT I O N
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1.1
DEPARTMENT
MANAGEMENT
STRUCTURE & STAFF
Director:
Prof. Francesco Nicotra
Vice Director:
Prof. Marina Lotti
Management Board:
Prof. Francesco Nicotra,
Prof. Antonio Zaza,
Prof. Enzo Martegani,
Prof. Giorgio Moro,
Prof. Luca De Gioia,
Prof. Silvia Barabino,
Dr. Paola Branduardi,
Dr. Anastasia Sguera.
Chief Financial Officer:
Dr. Anastasia Sguera
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FULL PROFESSORS
NAME
Alberghina Lilia
Castagnoli Paola
Fantucci Piercarlo
Lotti Marina
Lucchini Giovanna
FIELD
BIO/10
MED/04
CHIM/03
BIO/10
BIO/18
NAME
FIELD
Martegani Enzo
Nicotra Francesco
Ottolenghi Sergio
Porro Danilo
Tortora Paolo
BIO/11
CHIM/06
BIO/18
CHIM/11
BIO/10
NAME
FIELD
Giagnoni Gabriella
Grandori Rita
Granucci Francesca
Longhese Mariapia
Moro Giorgio
Nicolis Silvia
BIO/14
BIO/10
MED/04
BIO/18
CHIM/02
BIO/18
NAME
FIELD
Colombo Sonia
Combi Romina
Costa Barbara
De Filippis Lidia
Foti Maria
Fraschini Roberta
Frascotti Gianni
Fusi Paola
Galli Paolo
Gelain Fabrizio
BIO/11
BIO/13
BIO/14
BIO/13
MED/04
BIO/18
CHIM/11
BIO/10
BIO/07
BIO/13
NAME
FIELD
Vai Marina
Vanoni Marco
Wanke Enzo
Zaza Antonio
Zullini Aldo
BIO/11
BIO/10
BIO/09
BIO/09
BIO/05
ASSOCIATE PROFESSORS
NAME
Barabino Silvia
Becchetti Andrea
Cipolla Laura
Crosti Paolo
De Gioia Luca
Doglia Silvia Maria
FIELD
BIO/11
BIO/09
CHIM/06
BIO/01
CHIM/03
FIS/01
NAME
FIELD
Peri Francesco
Piatti Simonetta
Polissi Alessandra
Ronchi Antonella
Vescovi Angelo
CHIM/06
BIO/18
BIO/19
BIO/18
BIO/13
ASSISTANT PROFESSORS
NAME
Ambrosini Roberto
Bertini Luca
Brambilla Luca
Branduardi Paola
Brocca Stefania
Casiraghi Maurizio
Ceriani Michela
Chiaradonna F.
Coccetti Paola
Colangelo A.Maria
FIELD
BIO/07
CHIM/03
CHIM/11
CHIM/11
BIO/10
BIO/05
BIO/11
BIO/10
BIO/10
BIO/10
NAME
FIELD
Gorletta Tatiana
Labra Massimo
La Ferla Barbara
Lecchi Marzia
Orlandi Ivan
Regonesi Elena
Rocchetti Marcella
Tisi Renata
Zampella Giuseppe
MED/04
BIO/01
CHIM/06
BIO/09
BIO/11
BIO/10
BIO/09
BIO/11
CHIM/03
TECHNICAL AND ADMINISTRATIVE STAFF
Bottani Elena
Bruno Stefania
Campbell Neil
Colombo Marzia*
Comi Roberto
Delcarro Francesca
Gotti Maria Cristina
Lisini Alessandra *
Marchegiano Maria*
Mormile Bruno
Nanni Marco *
Sguera Anastasia
Smeraldi Carla*
Accardo Elena
Citterio Stefania
D’Urzo Annalisa
Farinaccio Antonella
Malerba Massimo
Marinoni Sara
Mostacciuolo Gaspare
Passolunghi Simone
Pedroni Paolo
Soler Valentina
Tonelli Maria Grazia
Urbano Matteo
Villa Anna Maria
Sacchetti Francesco
* temporary positions
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[6]
PhD STUDENTS
Acquati Serena
Alemanni Matteo
Amigoni Loredana
Aringhieri Chiara
Balestrieri Chiara
Bilotta Denis
Bodio Caterina
Bonetti Diego
Bosè Francesca
Bruni Ilaria
Busti Stefano
Caccia Roberta
Cantù Claudio
Casalgrandi Maura
Chisci Riccardo
Colombo Daniele
Comelli Francesca
Conforti A. Cristina
Consonni Silvia
Contran Nicla
Dato Laura
De Mattia Fabrizio
Di Domizio Alessandro
Di Resta Chiara
Falcetta Francesca
Farinaccio Antonella
Gaglio Daniela
Galati Elena
Galbusera Elena
Galliani Paolo
Gatti Lafranconi Pietro
Greco Claudio
Gregori Maria
Guerini Ilaria
Invernizzi Gaetano
Lancini Cesare
Losio Dario
Mantiero Davide
Mapelli Valeria
Marangoni Stefano
Mariani Jessica
Mazzantini Elisa
Mazzucchelli Serena
Merlini Laura
Metalli David
Orsato Alessandro
Ostuni Renato
Paiardi Chiara
Palmioli Alessandro
Panseri Silvia
Pasi Marco
Passolunghi Simone
Piazza Matteo
Piccirillo Sara
Pontiroli Francesca
Pozzi Chiara
Riva Matteo
Rossio Valentina
Santambrogio Carlo
Scandiuzzi Cristina
Schiavon Emanuele
Spreafico Roberto
Strona Giovanni
Taraballi Francesca
Torri Anna
Tosetti Valentina
Tripodi Farida
Venturetti Marianna
Vitali Caterina
Vivarelli Silvia
Zanda Valeria
Chiroli Elena
Clerici Michela
Codazzi Vera
Dante Mario
Del Favero Marta
Ferrari Daniela
Galimberti Andrea
Guaitolini Chiara
Lottersberger Francisca
Mainoldi Federica
Mastroianni Fabrizia
Mereghetti Paolo
Mezzasalma Valerio
Morini Raffaella
Natalello Antonino
Nwachukwu Joyce
Pagani Roberto
Panza Andrea
Papagna Angela Maria
Pastori Valentina
Piccirillo Sara
Ranghetti Anna
Redaelli Elisa
Rigamonti Valeria
Rossini Clara
Rota Nodari Laura
Rovelli Francesca
Soldati Chiara
Sommaruga Silvia
Sperandeo Paola
Spinosa Valerio
Stefani Fabrizio
Thurner Marina
Tsiarentsyeva Viktoria
Valotta Menella
Viganò Matteo
Villa Chiara
Viscardi Valeria
Vitullli Federico
Zarovni Natasha
Zolezzi Francesca
Favaro Rebecca
Ferri Anna Lucia
Fiocco Roberta
Fossati Tiziana
Gullo Francesca
Latorre Elisa
Lenzken Carolina
Mingozzi Francesca
Mortellaro Alessandra
Occhipinti Emanuela
Papaleo Elena
Redaelli Cristina
Restano Cassulini Rita
Sacco Elena
Samalikova Maria
Saracino Gloria
Splendiani Andrea
Zanoni Ivan
FELLOWS
Airoldi Cristina
Aracri Patrizia
Baldo Veronica
Barbuto Michela
Benzoni Francesca
Bettoni Isabella
Beretta Ottavio
Bigi Alessandra
Binda Elena
Bini Davide
Bonanno Davide
Caldarazzo Serena
Cassinelli Letizia
Chiarlone Anna Paola
POST-DOCS
Altomare Claudia
Baldissera Michela
Belotti Fiorella
Benzoni Francesca
Biancolini Donatella
Cirulli Claudia
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INTERNATIONAL MOBILITY 2007
INCOMING
from
group
Sofie Peeters
Katholieke Universiteit Leuven
(B)
Martegani
Feb-May 2007
Bert Vrancx
Katholieke Universiteit Leuven
(B)
Martegani
Feb-May 2007
Ana Catarina Araujo
Universidade de Lisboa, Lisboa
(P)
Nicotra
Jan-Mar 2007
Alexandre Orsato
Universidade Federal do Parana
Curitiba – Parana (Brasil)
Nicotra
From Nov 2007
Iwona Stawoska
Jagiellonian University Kraków
(PL)
Tortora
April-May 2007
OUTGOING
to
group
Pietro Gatti
Lafranconi
Universitat Autònoma de Barcelona
(E)
Lotti
Sept-Nov 2007
Maria Gregori
University of Southern Denmark,
Odense M (DK)
Nicotra
Sept-Nov 2007
Elena Galbusera
University Hospital of Geneva
Tortora
Oct 2007-Mar2008
Fabrizio Gelain
MIT Boston
(USA)
Vescovi
May and Nov 2007
Francesca Taraballi
A&M University, TEXAS
(USA)
Vescovi
May-Jun 2007
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1.2
ORGANIZATION
AND STRUCTURE
The Department
is organized as follows:
a. Managing Director’s Office
b. Administration Office
c. Students Office
d. Facilities and Assistance for
Teaching Activities
e. Technical Services
f. Software and Hardware
Assistance
g. Wastes Disposal Service
h. Workshop
i. Safety and Prevention Service
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a. MANAGING DIRECTOR’S OFFICE
Francesco Nicotra - Director; Francesca Loreto – Executive assistant
b. ADMINISTRATION OFFICE
Anastasia Sguera – Chief Financial Officer; Stefania Bruno - Foreign payments, VAT accounting; Roberto Comi Accounts Payable (Contracts, scholarships, travel); Francesca Delcarro - Purchase orders, Travel, Supplier’s
accounting; Bruno Mormile - Supplier’s accounting; Marco Nanni - Supplier’s accounting; Francesco Sacchetti Property inventory, Technical Attendance.
The Administrative Office works together with higher management to comply with all administrative aspects and procedures of the department management: arranges the Department and administrative board meetings, compiles
minutes and relays them to the appropriate office. The Office is responsible for general budget planning and financial reports; it manages the funds of the research groups and those dedicated to teaching activities; it manages relations with suppliers and external contractors
c. STUDENTS OFFICE
Maria Cristina Gotti, Elena Bottani, Marzia Colombo, Carla Smeraldi
The student administrative office manages all administrative aspects related to the teaching activities of the first level
degrees in Biotechnology and Biology and second level degrees in Industrial Biotechnology, Biology and Bioinformatics,
and of the Master in Bioinformatics. The student administrative office assists all students in the bureaucratic aspects of
their career; it is responsible for the content of the web pages of the department web site with regard to teaching activities; it organizes the calendar of lessons and exams and manages the data related to all degree courses through the
information system SIFA ON LINE.
d. FACILITIES AND ASSISTANCE FOR TEACHING ACTIVITIES
The Department hosts the following teaching laboratories
LABORATORY
PRACTICAL TEACHING ACTIVITIES
1011-1015 Chemistry Lab
Lab. of General and Inorganic Chemistry (BT)
Lab. of Organic Chemistry (BT)
Bio-Organic Chemistry (BT)
Purification and Downstream (BT)
Lab. of Chemistry (SB)
1026 Biochemistry Lab
Lab. of Biochemical Techniques (BT)
Lab. of Biomolecular Techniques (BT)
Molecular Pharmacology (BT)
1027-1029 Cell Biochemistry and Immunology Lab
Cell Biochemistry (BT)
Lab. of Immunological Techniques (BT)
Lab. of Cell Biochemistry Techniques (BT)
1028 Genetics and Microbiology Lab
Fermentation Technology (BT)
Lab. of Fermentative Techniques (BT)
Lab. of Genetic Techniques (BT)
Applied Microbiology (BT)
2010 Zoology and Comparative Anatomy Lab
Lab. of Experimental Biology (SB)
Zoology (SB)
2011 Experimental Biology Lab
Lab. of Experimental Biology (SB)
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2012 Experimental Biology Lab
Lab. of Experimental Biology (SB)
2013 Microscopy Lab
Fundamentals of Biology (SB)
Cytology and Histology (SB)
Plant Systems (SB)
Anatomy I (SB)
Anatomy II (SB)
2013 Microscopy Lab
Fundamentals of Biology (SB)
Cytology and Histology (SB)
Plant Systems (SB)
Anatomy I (SB)
Anatomy II (SB)
2029 Multimedia Lab
Supramolecular Chemistry (BT)
Physical Chemistry of Biological Systems (BT)
Bioinformatics: Basics (BT)
Bioinformatics: Structure-Function
Relationships (BT)
Molecular Biology (BT)
Numerical Methods for Bioinformatics (BT)
Fundamentals of Informatics (BT)
Computational Biochemistry (BT)
Immunogenomics (BT)
BT = Courses in Biotechnology
SB= Courses in Biology
e. TECHNICAL SERVICES
The technical staff is in charge of common services and maintainance of department or laboratory
instruments. The staff duties are as follows:
Elena Accardo
Chemistry lab 1011-1015, mass spectrometry
Stefania Citterio
Cytofluorimetry
Annalisa D’Urzo
Bioreactors Lab 2026, Biacore (plasmon resonance)
Antonella Farinaccio
Software technician, Lab 2029
Massimo Malerba
Lab of Morphological Microscopy
Sara Marinoni
Waste Disposal, Lab 1026
Gaspare Mostacciuolo
Technical gases (fluids), workshop
Simone Passolunghi
Fermentation technology Lab
Paolo Pedroni
Computer Manager
Valentina Soler
Microscopy Laboratory 2010-2013
Maria Grazia Tonelli
Genetic and Microbiology 1028
Matteo Urbano
Molecular Immunology 1027-1029, Liquid Nitrogen Service
Cooperation for Laboratory PL2 planning
Anna Maria Villa
Confocal Microscopy
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f. SOFTWARE AND HARDWARE ASSISTANCE
Paolo Pedroni
Services include: Departmental Server management, Database accounting management, Information technology departmental instruments management, Network of department management, Communication network management,
Accounts activation, Cooperation for computer lecture rooms management
g. WASTES DISPOSAL SERVICE
Sara Marinoni
The Service guarantees, according to law, disposal of chemical and biological wastes produced by the Department of
Biotechnologies and Biosciences and by the Department of Geological Sciences and Geotechnologies.
h. WORKSHOP
Gaspare Mostacciuolo
The Department includes a workshop for small mechanical repairs, for designing and making electronic instruments,
which may not be available commercially, for teaching and research uses. The workshop contributes to the maintenance and to the updating of equipment of teaching (Experimental Biology) and research laboratories (Cell Physiology).
i. SAFETY AND PREVENTION SERVICE
The service consists in following activities:
• maintaining contact with the Protection and Safety Office of the University
• collection, evaluation and filing of forms and news about Safety
• checking the correct working of security systems
• Cooperation for training and communication in safety issues
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1.3
INSTRUMENTATION
AND FACILITIES
A number of platform technologies and advanced instrumentations are available both to the
groups of the Department and
for scientific collaborations and
external services.
a. TECHNOLOGICAL PLATFORMS
BBC (BICOCCA BIOTECHNICUM CENTER)
A structure based on the most advanced scientific and
technological platforms required for upstream, bioreactor production, downstream and analysis of the product
(GMPlike env.). This structure is aimed at implementing
technological transfer through the development of biotechnological processes, pilot fermentations and research
contracts with companies as well as assistance in business plan preparation and patent writing and defence.
SYSTEMS BIOLOGY PLATFORM
Iterative integration of molecular and computational
approaches, aiming to structure genetic, biochemical
and genome-wide data (that produce detailed molecular descriptions of physiological and pathological pro-
cesses) so to foster the ability to comprehend and predict complex cellular functions.
TRANSCRIPTOMICS
Microarray technology allows for rapid measurement
and visualisation of differential expression between
genes at the whole genome scale. DNA microarrays, or
DNA chips, are fabricated by high-speed robotics,
generally on glass in which probes with known identity
are used to determine complementary binding, thus
allowing massively parallel gene expression and gene
discovery studies. Microarrays may be used to compare gene expression in two different cell types or tissue
samples, such as in healthy and diseased tissue.
Arrays are currently available for many biologically
relevant model organisms including human, mouse,
rat and arabidopsis.
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b. ADVANCED INSTRUMENTATION AND RESEARCH SERVICES
MASS SPECTROMETRY
The mass spectrometry (MS) facility at BtBs is equipped
with a quadrupole-time-of-flight (QSTAR elite, Applied
Biosystems) and a triple quadrupole/linear-ion-trap
(QTRAP, Applied Biosystems) instruments, for electrospray-ionization (ESI)-MS analysis and coupling to liquid
chromatography (LC). The QSTAR can operate in nanoESI also in connection with a nano-HPLC system. This
instrumentation allows peptide analysis by MS and
MS/MS for protein identification, sequencing and for analysis of posttranslational modifications. Furthermore, the
QSTAR machine allows for detection and characterization of intact proteins and protein non-covalent complexes for binding analysis and conformational studies.
BIOINFORMATICS AND MOLECULAR MODELLING
The Laboratory of Molecular Modelling provides high
performance computing facilities for Computer-Aided
Drug Design, computational methods in protein structure analysis and dynamics, bioinorganic quantum chemistry. Instruments available: High Performance Cluster :
15 blade Exadron Gbit LAN; each blade: 2 Intel Xeon 2.8
GHz, 512Mb RAM - HP proliant 4 opteron 2.4GHz, 4Gb HP proliant 4 opteron dual-core 2.8GHz, 8Gb RAMStorage unit Thecus, 2Tb
BIACORE X
Biacore uses Surface Plasmon Resonance (SPR) for in
real time label-free biomolecular interaction analysis
and generates unique data on molecular interactions
allowing to characterize molecules in terms of their specificity of interaction, on and off rates (kinetics), binding
strength (affinity) and providing accurate concentration
measurements. Major application areas include drug
discovery, general life science research, antibody characterization, immunogenicity, biotherapeutic development and manufacture.
NUCLEAR MAGNETIC RESONANCE (NMR) SPECTROMETRY
Structural characterization of molecules of smallmedum size (up to 5 kDa molecular weight). Proton and
carbon monodimensional and bidimensional experiments, gradient probe allowing 2D experiments in short
time (gCOSY, gTOCSY, gHMQC, gHSQC). Instruments
available: MERCURY 400 MHz, Varian. Software facilities
for advanced experiments (DOSY, Saturation Transfer
Difference, tr-NOESY).
CELL SORTING
The Flow Cytometry Core Facility provides instrumentation and expertise for automated cytofluorimetric analysis and sterile sorting of specific mammalian cell types.
The equipment of our facility consists of a MoFlo® high
speed cell sorter (Cytomation-BeckmanCoulter) equipped with three lasers (354 nm; 488 nm and 635 nm)
which enables to perform 9-colours analyses. The
MoFlo® has a dedicated operator who will help with
experimental design and running your samples.
MOLECULAR ANALYSES ON ANIMAL AND PLANT SAMPLES
DNA from animal and plant samples is sequenced and
analysed. Animal applications are in bird sex determination for monomorphic species and genotyping of animal
samples. The first issue allows for sex discrimination
based on sex specific PCRs, working on as few material
as a single feather in bird monomorphic species that do
not display clear morphological differences between
males and females. The second issue finds applications
in general activities of genotyping using PCRs, DNA
sequencing, DNA barcoding, microsatellites, SNPs.
Plant applications offer the possibility to characterize
plant by DNA markers. DNA, extracted from fresh or dry
plant materials, is analysed by microsatellites tools to
define the DNA fingerprinting and to evaluate the genetic relationship among close related species.
PHARMACOLOGY
The Pharmacology laboratory offers the following professional services: analgesic and antiinflammatory tests
in animal models, receptor binding assays, ELISA tests
for cytokines, hormons and transcription factors, cytotoxicity tests. Instruments available: Dynamic plantar
aesthesiometer, plantar test, plethysmometer, RandallSelitto analgesimeter, spectrophotometer, Liquid scintillation analyzer.
HPLC ANALYSIS
Analytical HPLC with gradient of eluents.
Semipreparative HPLC. Dual absorbance detection.
Quantitative analysis (Millennium and Borwin softwares).
Instruments available: Jasco (PU-2080 plus, UV-2075)
and Waters (515 pumps, 2487 dual absorbance UV)
instruments for analytical and semipreparative analysis.
EVALUATION OF MYOCARDIAL EXCITATION-CONTRACTION COUPLING
Expertise and tools for complete evaluation of myocardial
excitation-contraction coupling (electrophysiology, contractility and Ca2+ handling) provide a platform for the analysis of the mechanism of action of compounds developed
for cardiovascular therapy. Instruments available: patch
clamp units, video-edge detection of cell motion, intracellular Ca2+ measurement with high time-resolution, confocal analysis of “Ca2+ sparks”; Ca2+ imaging.
More information about instrumentation and costs of
analyses can be found on www.btbs.unimib.it
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1.4
T E AC H I N G ACT I V I T Y
BACHELOR IN BIOTECHNOLOGY
www.biotecnologie.unimib.it
The course lasts three years and is composed of a common two-year period, while the third year is devoted to
the following subjects: Industrial Biotechnology,
Molecular Biotechnology, Medical Biotechnology, this
last course carried out in cooperation with the Faculty of
Medicine and Surgery.
The overall number of enrolled students for the academic year 2006/2007 was 1.000
BACHELOR IN BIOLOGY
www.biologia.unimib.it
The graduation course is made up of a common year and a
two-year period devoted to the following subjects; bioecology, biomolecular and phyisiopathological studies.
The overall number of enrolled students for the academic year 2006/2007 was 850
MASTER IN BIOINFORMATICS
www.biotecnologie.unimib.it
The Master course in Bioinformatics is carried out over
two years in cooperation with the courses in Informatics
The overall number of enrolled students for the academic year 2006/2007 was 20
MASTER IN INDUSTRIAL BIOTECHNOLOGY
www.biotecnologie.unimib.it
The master course in Industrial Biotechnology is developed along two years and organized in the following areas:
1) Pharmaco-genomics and 2) Processes and Products.
The overall number of enrolled students for the academic year 2006/2007 was 170
MASTER IN BIOLOGY
www.biologia.unimib.it
The master course in Biology is developed along two
years and organized into the following areas: 1)
Functional and Molecular Biology 2) Bio-Ecology
The overall number of enrolled students for the academic year 2006/2007 was 70
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1.5
ADVANCED TRAINING
The Department hosts two
PhD programs in the frame of
the School of Doctorate in
Sciences (www.scuoladottorato.scienze.unimib.it), an international PhD program and a
master course.
PHD PROGRAM IN INDUSTRIAL BIOTCHNOLOGY
www.btbs.unimib.it
Coordinator: Prof. M. Vanoni
of the bio-medical research area and on the applications
that derive from the sequencing of complex genomes
such as of human and mouse.
The objective of the PhD in Industrial Biotechnology operative since the Academic Year 1999/2000, is to train
Researchers able to coordinate basic and applicative
research projects, in all fields of the biotechnological
industry. The training program plans a first preparatory
year and a two-year period devoted to one of the following subjects: biostructures, biosystems, bioprocesses
Moreover, members of BtBs participate in other PhD programs of the University of Milano-Bicocca and of the State
University of Milan: Physiology, Biochemistry, Chemistry,
Biomolecular and Genetic Sciences, Pharmacology,
Biology Applied to the evaluation an preservation of natural
resources, Nanotechnology.
PHD PROGRAM IN BIOLOGY
www.btbs.unimib.it
Coordinator: Prof. P. Tortora
The Biology Graduate Studies course is a complete cultural answer to the increasing demand for scientific and
professional training of young researchers able to operate with up to date technologies.
PHD PROGRAM IN TRANSLATIONAL
AND MOLECULAR MEDICINE
www.dimet.org
Coordinator: Prof. P. Castagnoli
DIMET is an international project supported by the programme Marie Curie and focuses on the recent advances
MASTER IN INTERNATIONAL
SCIENTIFIC NATURE SPECIALIST
www.isnweb.it
Coordinator: Dr. P. Galli
This Master’s course creates professional experts able to
merge both knowledge of marine natural environments
(Mediterranean and Tropics), communications, and teaching skills.
INTERNATIONAL MOBILITY: ERASMUS PROGRAM
Co-ordinator for Biology: Prof. Silvia Nicolis
Co-ordinator for Biotechnology: Prof. Marina Lotti
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SEMINARS 2007
26.1.2007
Anna Elisabetta Boccaccio
SISSA, Trieste
Meccanismi molecolari nella trasduzione olfattiva
29.1.2007
Shuguang Zhang
Massachusetts Institute of Technology (MIT)
Cambridge, MA, USA
Designer self-assembling peptides:
from repairing rat brains to harnessing
biosolar energy
5.2.2007
Cecilia Saccone
CNR – Istituto di Tecnologie Biomediche
Bari
Il genoma mitocondriale ed il suo ruolo come
marcatore di specie nei metazoi
I6.2.2007
Cecilia Saccone
CNR – Istituto di Tecnologie Biomediche
Bari
Duplicazione genica ed evoluzione delle
famiglie geniche OXPHOS nei vertebrati
9.2.2007
Roland Wohlgemuth
Sigma-Aldrich
Buchs, Switzerland
Enzymes in organic synthesis –
New tools and platforms
19 .2.2007
Andrea Tintori
Università di Milano
La durofagia tra i vertebrati marini
27.2.2007
Reiner Veitia
Université Denis Diderot/Paris VII
Polyalanines in health and disease
1.3.2007
Martino Bolognesi
Università di Milano
Basic principles and current high-throughput
applications in protein crystallography
9.3.2007
Piotr Chrzastowski-Wachtel
University of Warsaw, Poland
How to design information flow
in a structured way
21.3.2007
Salvador Ventura Zamora
Universitat Autònoma de Barcelona, Spain
Protein models to study folding,
aggregation and interactions
23.3.2007
Giovanni Malerba
Università degli studi di Verona
Studio della componente genetica
in malattie complesse
30.3.2007
Maria Luisa Rúa Rodríguez
University of Vigo, Spain
Lipolytic enzymes from Thermus thermophilus
HB27: from the plate to the structure
12.4.2007
Antonella Viola
Istituto Humanitas Milano
I recettori chemotattici nella sinapsi immunologica
17.4.2007
Wilfrida Decraemer
University og Gent, Belgium
The biogeography of plant parasitic Nematodes
20.4.2007
Valeria Wanke
Centre Médical Universitaire (CMU)
Geneve, Switzerland
La proteina kinasi Rim15 dirige l’entrata in G0
integrando segnali dalle vie PKA, TORC1
e Pho80-Pho85
20.4.2007
Silvia Brunelli
Università di Milano-Bicocca
and Institute San Raffaele, Milano
Wnt signalling in myogenesis
7.5.2007
Roberto Spagnoli
Sanofi Aventis, France
Metabolic engineering of baker yeast 1
8.5.2007
Roberto Spagnoli
Sanofi Aventis, France
Metabolic engineering of baker yeast 2
relazione 07
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[ 17 ]
SEMINARS 2007
9.5.2007
Roberto Spagnoli
Sanofi Aventis, France
Industrial specificities of bioconversions.
Traditional and cutting-edge approaches
10.5.2007
Roberto Spagnoli
Sanofi Aventis, France
Industrial specificities of biotechnology.
Production of proteins
14.5.2007
Simonetta Piatti
Università Milano-Bicocca
Regolazione della transizione
metafase-anafase negli eucarioti
15.5.2007
Andrea Musacchio
IFOM-IEO, Milano
Le basi molecolari del checkpoint mitotico
16.5.2007
Peter De Wulf
IFOM-IEO, Milano
Chromosome segregation in mitosis
and its regulation by the kinetochore
17.5.2007
Rosella Visintin
IFOM-IEO, Milano
Exit from mitosis in budding yeast Cdc14 promotes
its own return in the nucleolus through the
APC/C-Cdh1
18.5.2007
Roberta Fraschini
Università Milano-Bicocca
Regolazione dell'uscita dalla mitosi e citochinesi
da parte del checkpoint da posizionamento del fuso
18.5.2007
T. Achsel
IRCCS, Roma
Gli RNA messageri nei dentriti: implicazione per
la funzione neuronale del controllo del trasporto
nucleo-citoplasma, della stabilità e della traduzione
1.6.2007
Francesca Granucci
Università Milano-Bicocca
Induction and maintenance of T
cell tolerance in peripheral lymphoid organs
5.6.2007
S. Biffo
Università del Piemonte Orientale
Controllo della traduzione nel differenziamento
e nella trasformazione tumorale: principi ed esempi
8.6.2007
Nicoletta Landsberger
Università dell’Insubria
La sindrome di Rett, un paradigma dell'importanza
della metilazione del DNA per un corretto
funzionamento del Sistema Nervoso Centrale
12.6.2007
Ivan Zanoni
Università di Milano-Bicocca
Intercellular mechanism of DC-mediated NK
cell migration and activation
12.6.2007
Ernesto Gruccione
IEO, Milano
Myc and chromatin: a mutual relationship
15.6.2007
Tatiana Goletta
Università di Milano-Bicocca
Genetica dei tumori: studio della leucemia
mieloide acuta mediante tecniche high-throughput
18.6.2007
Andrea Morrione
Thomas Jefferson University
Growth factor signaling, receptor trafficking
and tumor development
19.6.2007
Alessandra Mortellaro
Università di Milano-Bicocca
Le immunodeficienze primarie: cause e terapie
19.6.2007
Gianmaria Maccaferri
Feinberg School of Medicine and Institute
of Neuroscience Northwestern University,
Chicago, USA
Tutto quello che avreste sempre voluto sapere
sugli interneuroni, ma non avete mai osato chiedere
21.6.2007
V. Zappavigna
Università di Modena
Le proteine Hox come fattori di trascrizione e non solo
relazione 07
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[ 18 ]
SEMINARS 2007
22.6.2007
A. Villa
Istituto Tecnologie Biomediche, CNR, Milano
Analisi dei meccanismi patogenetici della
Sindrome di Omenn
28.6.2007
Kristian Helin,
DIMET, Milano
Coordinated regulation of differentiation and cancer by
histone lysine methyl transferases and demethylases
3.7.2007
Luigi Casella
Università di Pavia
Biological oxidation and nitration by metalloproteins:
mechanistic aspects, biological effects and some
applications
7.9.2007
Enrico Rizzarelli
Università di Catania
Metalli e neurodegenerazioni
21.9.2007
Stephen Best
University of Melbourne
Chemical modeling of [FeFe]-Hydrogenase
24.9.2007
Lorenzo Lamberti
Milano
Le biotecnologie in Cina
25.9.2007
Lourival D. Possani
National Autonomous
University of Mexico Cuernavaca, Mexico
Toxins and venoms from scorpions:
medical and scientific aspects
19.10.2007
Lidia De Filippis
Università Milano-Bicocca
A novel, immortal, and multipotent human neural
stem cell line generating functional neurons and
oligodendrocytes
13.11.2007
Francesca Granucci
Università Milano-Bicocca
Intercellular and intracellular mechanisms
of dendritic cell-mediated NK cell activation
14.11.2007
Michael B. Hall
Department of Chemistry Texas
A &M University of Texas
Carbon-Hydrogen Bond Activation
21.11.2007
Maria Foti
Università Milano-Bicocca
Inflammatory signatures and molecular mechanisms
induced in Dendritic cells by microrganisms
27.11.2007
Elena Ghibaudi
Università degli Studi di Torino
Investigating protein unfolding and conformational
dynamics: an approach based on complementary
biophysical techniques and biochemical assays
29.11.2007
William Kelley
University Hospital of Geneva, Switzerland
From bacteria to viruses and man:
regulation of the Hsp70 chaperone by J-domain
partner proteins
4.12.2007
Frédéric Carrière
CNRS, Marseille, France
Pancreatic lipase: controlling substrate specificity
using an active site with a lid
6.12.2007
Sonia Longhi
CNRS et Universités d'Aix-Marseille I et II,
Marseille, France
Cartografia della transizione alfa-elicale della regione
intrinsecamente disordinata della nucleoproteina del
virus del morbillo tramite spettroscopia EPR
19.12.2007
Giovanni Cenci
Università dell'Aquila
Meccanismi di protezione dei telomeri in Drosophila
melanogaster
relazione 07
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14:31
Pagina 19
[ 19 ]
1.6
MASTER
AND PHD THESES
INDUSTRIAL BIOTECHNOLOGY
Annoni Emanuele “Ruolo della cisteina 121 nel processo di aggregazione in vivo e nella stabilità di beta-lattoglobulina bovina ricombinante”. M. Lotti
Buratti Claudia Laura Maria “Studio biochimico della
funzione citoplasmatica di Prep1 (pKnox1) nel controllo
della traduzione”. M. Vai
Apostolidis Emanuele “Proteine associate alla cisplatino resistenza. Analisi mediante tecnologie di linee cellulari di tumore dell’ovaio sensibili (2008) e resistenti
(2008/C13*) al cisplatino”. R. Grandori
Caldarazzo Serena Maria “Studio della stabilità di un
enzima adattato al freddo mediante strategie di evoluzione guidata”. M. Lotti
Avonto Cristina “Sviluppo di bioadditivi per l’industria
cartaria”. L. Brambilla
Callegarin Dario “Determination of ∆9-tetrahydrocannabinol (THC) effects on human breast cancer cell proliferation”. B. Costa
Balivo V “Sviluppo e caratterizzazione di mutanti ipersecretori per la produzione di proteine eterologhe in
Zygosaccharomyces bailii”. P. Branduardi
Calvi Francesca “Identificazione e caratterizzazione di
varianti iperattive della proteina di checkpoint Mec1 in
Saccharomyces cerevisiae”. M. P. Longhese
Bartesaghi Luca “Generazione di un modello murino
difettivo in distroglicano ed integrina ß1 nelle cellule di
schwann: ruolo nel “sorting” assonale”. E. Martegani
Bazzotti Riccardo “Il rimodellamento della superficie
cellulare in seguito ad esocitosi di enlargeosomi: specificità delle “shedding vesicles” rilasciate dalla membrana plasmatica”. E. Martegani
Caprioglio Manuela “Nuovi inibitori della proteina Ras
umana derivati dal monosaccaride D-glucosio”. F. Peri
Cardano Marina “Il coinvolgimento di SEL1 nello stress
del reticolo endoplasmatico”. E. Martegani
Castelnuovo Marilena “Caratterizzazione biochimica e
funzionale del prodotto del gene CLN8 associato ad una
forma di ceroidolipofuscinosi”. E. Martegani
Beretta Riccardo “Valutazione dell’aggressione del peptide beta-amiloide via Dynamic Light scattering e
MALDI-TOF”. R. Grandori
Cazzaniga Silvia “Studio in vitro dei meccanismi antinfiammatori dell’eparina. Possibili implicazioni per la terapia della malattia di Alzheimer”. F. Granucci
Bonfanti Katia “Analisi in vitro ed in silico dell’interazione recettore-ligando e valutazione del profilo di estrogenicità di composti di neo-sintesi”. P. Fantucci
Cerato Marco “Ricerca di un metodo per la determinazione quantitativa dell’Ammonio Metacrilato RL nelle
forme farmaceutiche retard: sviluppo di un nuovo
approccio analitico cromatografico e prime prove di
applicazione”. G. Frascotti
Braga Noemi “Metabolismo dell’acido ialuronico in
modelli in vitro di tumore della mammella”. R. Tisi
Brioschi Matteo “Analisi mediante stem-loop RT-PCR di
micro-RNA- ematopoietici su frazioni di mononucleate
nella leucemia mieloide acuta”. F. Granucci
Cimbro Raffaello “Analisi del loop V2 della proteina virale GP120 di HIV: costruzione di mutanti per lo studio della
possibile struttura assunta e per la futura creazione di un
vaccino”. M. Foti
relazione 07
23-04-2008
14:31
Pagina 20
[ 20 ]
Clementi Giorgia “Sintesi di multiple antigen peptide
come vaccini contro il virus dell’afta epizootica”. F. Peri
in seguito ad infezione con Listeria monocytogenes e
Listeria innocua”. M. Foti
Codazzi Vera “Espressione eterologa e purificazione di
proteine coinvolte nel controllo del ciclo cellulare del lievito Saccharomyces cerevisiae”. S. Brocca
Galati Elena “Ricerca di mutanti di lievito alterati nell’adattamento al checkpoint mitotico”. S. Piatti
Colleoni Lara “Ricerca mutazioni alleliche nel gene tiopurina-S-metiltransferasi (TPMT) in pazienti affetti da miastenia grave: correlazione tra genotipo e risposta al trattamento farmacologico con azatioprina”. B. Costa
Colombo Laura “Individuazione e determinazione di fattori di rischio biochimici e genetici coinvolti nella patogenesi della malattia aterosclerotica”. A. Becchetti
Cunietti Michela “Purificazione e caratterizzazione della
sticolisina I e di cinque mutanti non emolitici della sticolisina II”. B. Costa
D’ Amato Lisa “La policistina-1 induce migrazione cellulare regolando il riarrangiamento del citoscheletro attraverso PI3K e l’adesione meccanica cellula-cellula attraverso GSK-3ß. P. Coccetti
De Agostini Cristina “Sindrome da delezione 22q13.3:
identificazione e caratterizzazione molecolare in dodici
nuovi casi”. M. P. Longhese
Decio Alessandra Agnese “Role of CB2 receptors in neuroprotective effects of cannabinoids in animal models of
striatal degeneretion”. B. Costa
Del Mare Sara “Disegno razionale di peptidi antigenici
associati a tetrameri di HLA-DR1101 per l’identificazione
di linfociti T CD4+ antitumorali”. F. Granucci
Di Vona Chiara “Funzione dell’ubiquitino ligasi cop1 nella
risposta all’irraggiamento UV”. G. Lucchini
Donelli Ilaria “Studio della modificazione superficiale del
poli-(etilen tereftalato) mediante impiego di enzimi lipolitici”. R. Grandori
Fenaroli Federico “Il trapianto di cellule staminali mesenchimali quale approccio terapeutico al dolore neuropatico”. G. Giagnoni
Fialà Stefano “Studio di attività e stabilità di BaeyerVilliger monoossigenasi in miscele acqua-solvente organico”. M. Lotti
Fracasso Valentina “Caratterizzazione funzionale
della proteina coinvolta nella patogenesi di una nuova
forma di atassia spinocerebellare dominante: SCA28”.
L. Alberghina
Fumagalli Elisabetta “Effetti dell’espressione del gene
Osmyb4 sul metabolismo primario e secondario in diverse specie vegetali”. M. Vai
Fumagalli Silvia “Studio dell’interazione tra cellule dendritiche e batteri: analisi globale dell’espressione genica
Galioto Maria Eleonora “Caratterizzazione del ruolo delle
proteine Dma di lievito nel controllo della dinamica dell’anello di septine e della citochinesi”. S. Piatti
Giudici Jacopo “Studi di localizzazione di Ras2-GTP nel
lievito Saccharomyces cerevisiae”. E. Martegani
Isella Francesca “Studio del Grafting di substrati cheratinici con transglutaminasi microbica (Streptoverticillum
mobaraense)”. M. Vanoni
Isolato Marco “Sviluppo di un metodo per la microstampa
di proteine di interesse farmacologico”. M. Vanoni
Issi Luca “Caratterizzazione chimica ed enzimatica di
galattomannanni da Cyamopsis tetragonolobus per applicazioni industriali ”. M. Vanoni
Jirillo Giuseppe “Analisi delle alterazioni genomiche nell’adenocarcinoma polmonare”. G. Lucchini
Jocollè Genny “La melanocortina sintetica (CKPV)2 come
terapia innovativa della flogosi”. F. Nicotra
Lattanzi Marco “Studio della fisiologia di Saccharomyces
cerevisiae in coltura continua limitata da azoto”. L.
Brambilla
Libri Domenico Vladimiro “Sviluppo e caratterizzazione di
un anticorpo diretto contro C-terminale di SEL1L”. E.
Martegani
Livrieri Silvio “Immobilizzazione della fenilacetone monoossigenasi e sue applicazioni nella sintesi organica asimmetrica”. F. Nicotra
Locatelli Giuseppe “Ruolo della proteina adattatrice Rai
(ShcC) in cellule staminali tumorali estratte da glioblastomi”. F. Chiaradonna
Manfrini Nicola “Ruolo della protein chinasi di S. cerevisiae Rad53 nell’attivazione del checkpoint durante il ciclo
cellulare meiotico”. M. P. Longhese
Marcassoli Filippo “Una singola traslocazione BIT VIIIXV provoca l’alterata espressione di molte proteine
trasformando S. cerevisiae in una cellula gravemente
aberrante”. D. Porro
Medina Filippo “Studio del ruolo delle cellule dendritiche
nell’attivazione delle funzioni antitumorali delle cellule
NK”. F. Granucci
Mercuri Dario “Studio dei cambiamenti conformazionali indotti da ligando della D-aminoacido ossidasi
umana”. R. Grandori
relazione 07
23-04-2008
14:31
Pagina 21
[ 21 ]
Modica Livia “Materiali auto-assemblanti per la rigenerazione del tessuto nervoso periferico”. A. Vescovi
e tossicologico di inibitori chinasici su meccanismi di
segnalazione intracellulare”. P. Coccetti
Montana Emanuela “Sviluppo di un biocatalizzatore per la
desacetilazione di cefalosporine semisintetiche”. P.
Branduardi
Rossini Clara “Sintesi di nuovi antagonisti del lipide A attivi contro lo shock settico, l’ischemia e il dolore neuropatico”. F. Peri
Orrù Roberto “Modulazione dell’affinità per l’ossigeno
della D-aminoacido ossidasi mediante irrational
design”. M. Lotti
Rovelli Elisabetta “Caratterizzazione funzionale della ferritina mitocondriale espressa in cellule HeLa e in fibroblasti di pazienti con atassia di Friederich”. M. Vanoni
Ostuni Renato “Definizione del pathway di trasduzione del
segnale indotto dal legame di LPS liscio al complesso
recettoriale TLR4/CD14”. F. Granucci
Sala Simone Giovanni “La simvastatina riduce la proliferazione indotta da terreno in linfociti immortalizzati con
EBV, provenienti da pazienti affetti dal morbo di
Alzheimer, indipendentemente dagli effetti conseguenti
l’abbassamento del livello di colesterolo”. E. Martegani
Palazzotto Floriana “Meccanismi molecolari responsabili
dell’azione promigratoria dell’ossitocina in cellule endoteliali umane”. G. Giagnoni
Pellegatta Marta “Il ruolo di Neurofibromina, il prodotto
del gene NF1, nell’attivazione di Ras ed ERK: un’analisi in
vitro ed in vivo”. E. Martegani
Perrella Marco “Modulazione nicotinica delle correnti
post-sinaptiche inibitorie nella corteccia prefrontale di
topo”. A. Becchetti
Pianaroli Allegra “Studio del ruolo del gene RaLP nella
differenziazione embrionale murina”. M. Vanoni
Piccolo Francesco “Caratterizzazione della risposta
immunitaria a carico dei linfociti T che riconoscono molecole CD1 associate ad antigeni endogeni”. F. Granucci
Pileggi Silvana “Valutazione del contributo degli aplotipi
del citocromo P450 (2C9) e di VKORC1 nel dosaggio del
farmaco nella terapia anticoagulante” E. Martegani
Protta Giulia “Strategie di inibizione/antagonismo di
HMGB1, una chemochina pro-infiamatoria coinvolta nell’induzione e/o nel mantenimento di gravi patologie
umane”. M. Vanoni
Razini Paola “Studio delle capacità proliferative e di differenziamento mio-endoteliale delle cellule staminali
CD133+: nuove prospettive terapeutiche nella distrofia
muscolare di Duchenne”. B. Costa
Rigamonti Nicolò “Espressione intracellulare di uno specifico Affibody per inibire la funzionalità di HER-1,
mediante ritenzione nel reticolo endoplasmatico”. M.
Lotti
Ronchi Carlo Luigi “Ruolo della miosina6 nella regolazione del segnale e del traffico intracellulare del
recettore dell’EGF”. S. Piatti
Rossi Giorgia “Integrazione di approcci computazionali e
di ingegneria metabolica volti alla modulazione del flusso
glicolitico in Saccharomyces cerevisiae”. P. Branduardi
Rossi Paola Maria “Valutazione dell’effetto farmacologico
Saracino Filippo “Farmacocinetica del gimatecan, una
nuova captotecina orale, in pazienti con sarcoma di tessuti molli, in sperimentazione clinica di fase II”. F. Nicotra
Sartori Matteo Francesco “Caratterizzazione e differenziamento di cellule staminali adulte umane isolate da tessuto adiposo sottocutaneo per l’impiego in ingegneria tissutale”. G. Giagnoni
Savoldi Boles Michela “Ingegnerizzazione di mutanti di
batteri rossi non sulfurei per lo sviluppo di processi fermentativi idonei alla produzione di H2”. P. Tortora
Scalcinati Gionata “Metabolic engineering of Saccharomyces
cerevisiae for biochemical production from xylose”. D.
Porro
Sedini Valentina “Sviluppo di microsistemi analitici
per la rivelazione di microorganismi patogeni in ambito alimentare”. I. Orlandi
Serra Domenico Maria “Studio sugli effetti degli acidi
grassi polinsaturi n-3 (n-3 PUFA) su cardiomiociociti isolati di ratto esposti a danno da ipossia/riossigenazione”.
A. Zaza
Simonetti Elena “Caratterizzazione dell’enzima responsabile della sintesi di cADPR e di NAADP” B. Costa
Solaroli Michele “Purificazione e caratterizzazione di
mutanti deglicosilati di lipasi ricombinante da Candida
rugosa” S. Brocca
Somaschini Alessio “Degradazione proteolitica della
atassina-3: analisi dei frammenti e della loro localizzazione sub-cellulare”. P. Fusi
Tagliabò Claudia “Fluorescence analysis of the immobilized Lactobacillus brevis alcohol dehydrogenase used in
the gas phase reaction”. L. Cipolla
Thurner Marina Daniela Francesca “Realizzazione di
sistemi ad alta efficienza per l’espressione di proteine di
membrana ricombinanti”. M. Lotti
relazione 07
23-04-2008
14:31
Pagina 22
[ 22 ]
Tomelli Marina “Veicolazione di antigeni vaccinali
mediante nanosfere polimeriche biocompatibili” F. Peri
Tumaini Barbara “Studio del ruolo di uPAR nella progressione neoplastica: possibile cooperazione di questo recettore con i due oncosoppressori murini p16INK4a e
p19ARF”. G. Lucchini
Uccellini Lorenzo “Studio del ruolo del sistema IL7/IL7R,
della citochina IL15 e dei polimorfismi dei recettori di IL7
e di IL15 nella progressione dell’infezione da HIV-1”. G.
Lucchini
Vaghi Valentina “Effetti anticonvulsivanti mediati dall’espressione del gene del neuropeptide Y (NPY) attraverso l’utilizzo di vettori virali adenoassociati” B. Costa
Viganò Eleonora Maria “Caratterizzazione degli effetti
farmacologici di alcuni Growth Hormone Secretagogues”.
G. Giagnoni
Zanda Valeria Maria “Studi sul targeting di farmaci antitumorali”. B. La Ferla
BIOLOGY
Altara Raffaele “Clonaggio e caratterizzazione molecolare del repertorio autoanticorpale di isotipo IGM di una
paziente HCV-positiva con crioglobulinemia”. A. Polissi
Ambrosi Paola “Proprietà funzionali del recettore nicotinico alfa2-I279N-beta4, implicato in forme di epilessia
notturna”. A. Becchetti
Aquaro Giovanni “Nuovi approcci metodologici per lo studio di un sistema ospite parassita”. P. Galli
Asnaghi Valentina “Monitoraggio dei popolamenti superficiali delle coste rocciose liguri: qualità ecologica ai sensi
della water framework directive 2000/60/EC”. P. Galli
Bodio Caterina ”Ruolo delle proteine ELAV nella regolazione post-trascrizionale del gene Nova1 nel modello
motoneuronale NSC-34”. S. Barabino
Bonandrini Barbara “Effetto dell’antagonista del recettore cannabico CB1, Rimonabant, in un modello murino di
neuropatia diabetica”. G. Giagnoni
Buscarino Michela “Filogeografia di Austropotamobius
pallipes (Decapoda, astacidae) nel bacino padano”. P. Galli
Caniatti Giacomo Upendo “Sbiancamento (bleaching) dei
coralli: un approccio ecologico e molecolare”. M.
Casiraghi
Delai Stefania “Un innovativo approccio farmacologico per
il trattamento del dolore neuropatico: gli antagonisti del
Toll-like receptor 4 come modulatori della microglia attivata”. B. Costa
D’Errico Paolo “Atrofia muscolare spinale (SMA):
caratterizzazione morfologica ed espressione delle
proteine smn in motoneuroni di pazienti affetti e di
topi transgenici”. S. Ottolenghi
Di Virgilio Silvia “Analisi filogeografica di Cladocora
caespitosa (L., 1767) nel Mediterraneo settentrionale”.
P. Galli
Donadelli Silvia “Uno studio sui nematodi crenofili del
Trentino”. A. Zullini
Donadoni Carla “Risposte immunitarie mucosali in
individui esposti all’infezione dell’HIV-1 ma non infetti”.
F. Granucci
Dose Alice “La teoria delle reti applicata allo studio delle
simbiosi”. M. Casiraghi
Ferrario Filippo “Biodiversità e distribuzione della
Spongofauna del parco marino di Ras Mohammed,
Egitto”. P. Galli
Fontana G “Studio della regolazione dell’espressione di
EFA6As in cellule neuronali in cultura”. S. Barabino
Galimberti Andrea “Dilemma Tassonomico in specie
alloctone invasive: il caso di Paradoxornis webbianus E P.
alphonsianus nella riserva naturale palude brabbia (VA)”.
M. Casiraghi
Gallina Enrico “Autoecologia di Silurus glanis nel lago di
Comabbio (VA)”. P. Galli
Gugerotti Laura “Diagnostica di laboratorio della malattia
tubercolare: esame microscopico, esame colturale e biologia molecolare”. A. Polissi
Ierardi Rossella ”Il ruolo del fattore trascrizionale EBF2
nella formazione delle circuiterie cerebellari”. S. Nicolis
Longhin Eleonora “Ciclo riproduttivo di Cladocora caespitosa (Scleractinia, Faviidae) nel Mar Ligure”. P. Galli
Maggioni Melissa “Ricerca e caratterizzazione degli interattori della proteina Bert in cellule staminali embrionali
murine”. S. Nicolis
Magni Giorgio Federico “Caratterizzazione delle biocenosi nell’area del Parco Marino di Watamu, Kenya”. P. Galli
Castagnaro Laura “Cellule di pazienti Down e murine
overesprimenti il fattore trascrizionale Prep-1 presentano
un incremento di apoptosi p53 dipendente”. S. Nicolis
Marangoni Stefano “Cellule staminali cardiache murine:
analisi molecolare e funzionale”. A. Zaza
Colombo Federica “Struttura di sistemi ospite-parassiti
alloctoni”. P. Galli
Mauri Mario “Espressione e localizzazione dei recettori
alfa e beta per il trombossano”. G. Giagnoni
relazione 07
23-04-2008
14:31
Pagina 23
[ 23 ]
Scaccabarozzi Daniela “La vegetazione della Valsassina –
storia olocenica e fasce altitudinali attuali. Analisi pollinica dei depositi lacustri di barzio”. P. Crosti
Segrè Valentina “Il ruolo della SUMOrilazione nei complessi di regolazione trascrizionale contenenti deacetilasi
istoniche 1 e 2”. S. Ottolenghi
Siesa Matteo Elio “Il genere Lucanus Scopoli, 1763
(Coleoptera Lucanide) nella fauna italiana: aspetti molecolari, morfologici ed ecologici”. M. Casiraghi
Tomasin Elisa “Incongruenze filogenetiche nella sistematica dei coralli (Cnidaria: scleractinia): il caso dei
Siderastreidae”. P. Galli
Vallebona Sara “Ruolo delle sinapsi elettriche e delle correnti di “pacemaker” in reti di neuroni corticali di topo”. E.
Wanke
Vismara Elena Maria Carlotta “Identificazione di partner
molecolari dell’atassina-3”. P. Tortora
Zulian Alessandra “Refrattarietà alla neurodegenerazione calcio-dipendente e aumento della longevità nei topi
knockout per Surf1”. S. Ottolenghi
BIOINFORMATICS
Bossi Alice “Data mining su dati molecolari per il drug
discovery”. P. Fantucci
Cozzi Alessandro “Sviluppo di un database per la validazione di procedure di correlazione tra funzionalità proteiche e superfici molecolari”. L. De Gioia
Di Leo Giuseppe “Studio computazionale di composti
attivi del tamoxifene sui recettori degli estrogeni
(E.R.)”. P. Fantucci
Negroni Jacopo “Sviluppo ed automatizzazione di un protocollo bioinformatico per l’analisi dell’interazione proteina-ligando con calcolo ad alta prestazione”. G. Moro
Riccardi Laura “Protein dynamics, in silico mutagenesis,
and unfolding: a molecular dynamics approach of coldadapted serine-proteases”. L. De Gioia
Visconti Marta “Simulazione dell’interazione della catena
leggera del fattor V di coagulazione con una membrana
fossolipidica mediante dinamica molecolare”. P. Fantucci
OTHER
Santambrogio Carlo “Transizioni conformazionali e formazione di complessi non covalenti di proteine analizzate
tramite nano-ESI MS”. R. Grandori. Degree in Physics
PhD DISSERTATIONS
Bilotta Denis “Study of structure-function relationship in
proteins via comparative molecular dynamics simulations: the case of SpeB”. PhD in Industrial Biotechnology.
Tutor: P. Fantucci
Bonanno Cinzia “Development of a novel real-time PCR
technology (MS-FLAG) for the detection of aberrant
methylation in gene promoters”. PhD in Industrial
Biotechnology. Tutor M. Vai
Dato Laura “Yeast stress in biotechnological processes:
stress response analysis and strain improvement strategies”. PhD in Industrial Biotechnology. Tutor: P.
Branduardi
Gatti Lafranconi Pietro “Cold-adaptation, thermal stability and in vivo aggregation of a lipolytic enzyme”. PhD in
Industrial Biotechnology. Tutor: M. Lotti
Greco Claudio “A DFT and QM/MM investigation on
models related to the [FeFe]-hydrogenase active site”.
PhD in Chemistry. Tutor: L. De Gioia
Invernizzi Gaetano “In vitro and in vivo studies on the conformational stability of model proteins”. PhD in Industrial
Biotechnology. Tutor: M. Lotti
Losio Dario “Correlation among cultural condition, metabolism and physiology in Saccharomyces cerevisiae“. PhD
in Industrial Biotechnology. Tutor: L. Brambilla.
Mantiero Davide “Tel1/ATM and Sae2/CtIP govern different aspects of the DNA damage response”. PhD in
Biology. Tutor: Giovanna Lucchini.
Mingozzi Francesca “Studio dei meccanismi di induzione
di tolleranza in cellule T autoreattive da parte di diverse
APC negli organi linfoidi periferici”. PhD in Immunology,
University of Rome. Tutor: F. Granucci
Paiardi Chiara “Studies on Ras/Cyr1/Cdc25 signalling
complex localization and evidences for Cyr1 as scaffold
protein for Ras2-Ira interaction in Saccharomyces cerevisiae”. PhD in Industrial Biotechnology. Tutor: E.
Martegani.
Scandiuzzi Cristina “The NGF specific receptor TrkA:
regulation of turnover and downstream pathway”. PhD in
Industrial Biotechnology. Tutor: E. Martegani.
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2
R ESEARCH
G ROUPS
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1
MOLECULAR GENETICS
OF DEVELOPMENT
AND CELL DIFFERENTIATION
IN MOUSE AND MAN
PARTICIPANTS
Silvia Nicolis, Sergio Ottolenghi,
Antonella Ronchi, Rebecca Favaro, Anna Ferri.
The development of complex organs and tissues, such as
brain and the hematopoietic system, requires the ordered
expression of key transcription factors controlling cell
type- and tissue-specific gene expression. Stem cells
represent the self renewing compartment of rapidly replicating cell types, as in the hematopoietic system, but are
present, in small numbers, also in adult brain, heart and
other organs which do not show active cell replication in
adults. The group uses a common set of approaches (conditional and standard targeted mutagenesis in mouse,
cell culture and gene transduction, chromatin studies,
etc.) to investigate the role of key transcription factors in
the development, maintenance and differentiation of a
variety of stem cells.
ter gene in Primordial Germ Cells, and some of their
descendants, as well as in Hematopoietic Stem Cells and
early progenitors. Using Chromatin Immunoprecipitation
and other techniques (3C assay) we are trying to define
transcription factors interacting with the main regulatory
areas of the gene (with B. Gottgens, Cambridge). The
regulation of the same transgene is also being studied in
embryonic stem cells and in spermatogonia (with S. Dolci,
Univ. Roma 2). GFP expression driven by the same transgene is also used to track stem cells migrating from the
bone marrow to the heart, and to test their ability to develop in vitro as cardiac stem cells (with E. Messina, Roma
and C. Magli, CNR, Pisa).
THE ROLE OF SOX2 IN STEM CELLS.
THE ROLE OF SOX AND OTHER TRANSCRIPTION
FACTORS IN HEMATOPOIETIC DEVELOPMENT AND
GLOBIN REGULATION.
S.Nicolis, R.Favaro, A.Ferri, C.Lancini, E.Latorre, J.Mariani, R.Caccia, V.Tosetti.
Sox2 is a transcription factor critically involved in multipotency. Using Cre-mediated conditional ablation of Sox2, in
vivo and in vitro, the group is investigating the mechanisms of Sox2-dependent regulation of the development
of Embryonic and Neural Stem cells, and of their neuronal differentiation. Targets of Sox2, required for these
activities, are being identified by genomic and proteomic
studies, and by Chromatin Immunoprecipitation and 3C
conformation assays. Their functional role is then validated by in vitro lentiviral overexpression of the identified
genes. Collaboration on these topics is ongoing with
N.Dillon (Londra) and A.Smith (Cambridge, UK).
Additional aspects of the role of Sox2, in other stem cell
types, such as germ or skin cells, are being studied with
S. Dolci (Univ. Roma 2) and F. Watt (Cambridge).
MOLECULAR REGULATION OF THE c-KIT GENE
S.Ottolenghi, L.Cassinelli.
Using transgenic constructs, we identified a subset of ckit genomic sequences which drive expression of a repor-
A.Ronchi, S.Ottolenghi, C.Cantù, M.Casalgrandi, M.Baldissera.
We previously identified a large sets of genes which are
differentially expressed during the development of the
mouse hematopoietic system and its initial differentiation
(in fetal liver). We are presently focusing our work on a
number of differentially expressed transcription factors,
among which are some Sox-family factors, as well as on
some molecules (GATA-1 and GATA-2, CP2) already
known to control the expression of globins and other erythroid specific genes. By in vitro transfection, in vitro protein interaction studies, Chromatin Immunoprecipitation
assays, proteomic analysis and lentiviral transduction in
primary mouse and human hematopoietic cells, we are
trying to identify relevant functional targets of these transcription factors and to assess their effects on proliferation, differentiation and, for some of them, on the regulation of embryonic, fetal and adult globin synthesis.
Collaborations are with M.D. Cappellini (Univ. di Milano),
M. Ruoppolo (Napoli) and T. Enver in Oxford, UK.
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MECHANISMS OF
POST-TRANSCRIPTIONAL
REGULATION OF MAMMALIAN
GENE EXPRESSION AND THEIR
ROLE IN HUMAN DISEASE
2
PARTICIPANTS
Silvia M.L. Barabino, Carolina Lenzken,
Chiara Aringhieri, Silvia Vivarelli, Davide Bonanno,
Simona Paro, Reinaldo Alvarez.
Our Laboratory is interested in decoding the mechanisms involved in biogenesis of messenger RNA (mRNA)
and in contributing to a better understanding of the
pathogenesis of diseases caused by deregulation of this
process. Eukaryotic messenger RNA precursors (premRNAs) are synthesized and processed in the nucleus
prior to their export to the cytoplasm, where they serve
as templates for protein synthesis. Transcription is coupled spatially and temporally to capping of the premRNA at the 5’ end, to splicing of introns and to 3’ end
polyadenylation. Very little is known about how these
events are spatially and temporally coordinated within
the context of the nucleus to allow for the regulation of
accurate gene expression.
3’ END PROCESSING AND TRANSPORT OF mRNAs
Although stimulation of mRNA export by polyadenylation
has long been observed, so far little is known about the
possible mechanism. We have characterized the intracellular localization of the 3’ end processing factor CF Im
and we have shown that it shuttles continuously between the nucleus and the cytoplasm in association to
mRNA. Nucleo-cytoplasmic shuttling reflects its role in
coupling mRNA processing to later events in the life of
mRNA. We have found that CF Im68 interacts with the
major mRNA export receptor in vivo and in vitro.
Silencing of CF Im68 expression by RNAi results in the
accumulation of mRNAs in the nucleus. Our findings
argue for a direct involvement of CF Im68 in mRNA
export as a novel adapter protein for mRNA export factors.
CELL STRESS AND RNA SPLICING
Coupling of pre-mRNA splicing to extracellular signals
is crucial for altering splicing patterns according to the
physiological state of cells. Since protein phosphorylation is often the response of cells to external signals, our
working hypothesis is that phosphorylation-dependent
signal transduction cascades will ultimately regulate
alternative splicing pathways. We have recently established a cellular model that will allow us to elucidate the
molecular changes in the alternative splicing machinery
induced by the cellular stress response. In collaboration
with the Genopolis consortium we are using splicingsensitive microarrays to analyze changes in the expression of alternative mRNA isoforms in a cellular model of
neurodegenration.
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CONTROL OF GENETIC
INTEGRITY BY THE DNA DAMAGE
CHECKPOINT PATHWAYS
3
PARTICIPANTS
Maria Pia Longhese, Giovanna Lucchini, Michela
Clerici, Veronica Baldo, Valeria Viscardi,
Diego Bonetti, Davide Mantiero, Ilaria Guerini,
Nicola Manfrini and Francesca Calvi.
The genome of living organisms can suffer both spontaneous and induced DNA damage. DNA double-strand
breaks (DSBs) are among the most deleterious types of
damage that can occur in the genome of eukaryotic
cells, because failure to repair these lesions can lead to
genetic instability. Eukaryotic cells have to cope with
three different types of DSBs: accidental DSBs, programmed DSBs and natural DSBs. Accidental DSBs can
arise during both mitosis and meiosis of eukaryotic cells
either by DNA replication problems or by exposure to
environmental factors, whereas site-specific DSBs are
introduced into the genome in a programmed manner to
initiate meiotic recombination in germ cells. Finally,
eukaryotic cells contain natural DSBs that are represented by the ends of their linear chromosomes.
The cellular response to either accidental or programmed DSBs requires highly conserved surveillance
pathways, called DNA damage checkpoint and recombination checkpoint, respectively, which delay mitotic and
meiotic cell cycle progression until DSBs are repaired.
Mechanistically, the DNA damage checkpoint is related
to the recombination checkpoint. In fact, highly conserved protein kinases, including mammalian ataxia telangiectasia mutated (ATM) and ataxia telangiectasia and
RAD3-related (ATR), as well as their S. cerevisiae orthologs Tel1 and Mec1, are necessary to activate both the
DNA damage and the recombination checkpoint. Not
surprisingly, defects in these networks result in a variety of diseases ranging from severe genetic disorders to
cancer predisposition and accelerated aging.
In contrast to accidental and programmed DSBs, the
physical ends of eukaryotic chromosomes are protected
from checkpoints and other events that normally promote DSB repair. This differentiation is thought to be the
consequence of a unique organization of chromosomal
ends into specialized nucleoprotein complexes called
telomeres. When chromosome end protection fails,
dysfunctional telomeres are targeted by the DNA repair
and recombination pathways. The outcomes of such
events at telomeres range from the generation of chromosomal abnormalities, general hallmarks for cancer
cells in humans, to permanent cell cycle arrest and cell
death. Given the different fates of DSBs and telomeres, it
is remarkable that Tel1/ATM and Mec1/ATR are telomere-associated and are involved in ensuring telomere
length and identity, implying that the difference between
a DNA break and a telomere is less pronounced than
previously assumed. Our research project aims to elucidate the molecular mechanisms protecting telomeric
ends and controlling the cellular response to DSBs
during both the mitotic and meiotic cell cycles. In particular, we are using different approaches in order to provide new insights into the roles of ATM/Tel1 and
ATR/Mec1 checkpoint kinases in sensing, processing
and signalling mitotic and meiotic DSBs and telomeres.
Moreover, we are searching for new molecular targets of
these kinases and we are studying how these mechanisms are coupled to cell cycle progression and interconnected with each other.
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MITOTIC PROCESSES
PREVENTING GENOME INSTABILITY
AND ANEUPLOIDY
4
PARTICIPANTS
Roberta Fraschini, Elena Chiroli, Valentina Rossio,
Marianna Venturetti, Laura Merlini, Elena Galati, Ilaria Catusi,
Giovanna Lucchini and Simonetta Piatti.
Genetic instability involves gain or loss of genetic information and is thought to be one of the major causes of cancer
development. An altered chromosome number, referred to
as aneuploidy, is a hallmark of cancer cells. Mistakes
during mitosis may be responsible for the abnormal
karyotypes of many human tumour cells and have an
important role in oncogenesis.
The integrity of the genome depends upon surveillance
mechanisms, or checkpoints, which monitor the completion of critical cell cycle events and delay cell cycle progression until errors have been corrected. Thus, these
control mechanisms ensure the genetic stability of a cell’s
lineage. Checkpoint defects can pave the road to chromosome alterations and, ultimately, to cancer. Similarly,
recent findings indicate that human cells undergoing a
faulty cytokinesis accumulate numerical and structural
chromosome aberrations, presumably due to the formation of multipolar spindles. Thus, cytokinesis needs to be
tightly regulated in order to avoid aneuploidy. Our group
studies these issues using the budding yeast S. cerevisiae
as model organism. In particular, we are focusing on three
main research topics:
1. REGULATION OF MITOTIC PROGRESSION BY THE
SPINDLE ASSEMBLY CHECKPOINT.
Once mitotic chromosomes are duplicated into two sister
chromatids, their segregation is mediated by a bipolar
microtubule spindle, to which they attach via their kinetochores. When the sister kinetochores of each chromatid
pair are captured by microtubules emanating from opposite spindle poles, the chromosome becomes bi-oriented.
Finally, the onset of anaphase, where sister chromatids
split and migrate to the spindle poles, is one of the major
points of no return in the cell cycle, and unbalanced chromosome segregation at this stage will inevitably result in
the production of aneuploid cells. Therefore, anaphase
must be kept under check and delayed until all chromosomes are bi-oriented, a task carried out by the spindle
assembly checkpoint (SAC). In case of errors, the SAC
sends an inhibitory signal that delays the separation of
sister chromatids and mitotic exit until bipolar attachment
is achieved. The target of the SAC is the Cdc20/APC ubiquitin ligase, which is normally required for sister chromatid
separation and mitotic exit. We study some aspects of SAC
activation and switch-off in yeast.
2. CONTROL OF MITOTIC EXIT AND CYTOKINESIS BY THE
SPINDLE POSITION CHECKPOINT
In most eukaryotic cells the site where cytokinesis takes
place is dictated by the position of the mitotic spindle. In
budding yeast, conversely, the site of cell division, the bud
neck, is established already at the G1/S transition, concomitantly with bud emergence and much earlier than bipolar spindle formation. A surveillance mechanism called
spindle position checkpoint delays cytokinesis in the presence of misoriented spindles. The spindle position checkpoint operates through down regulation of the small
GTPase Tem1, acting at the top of the mitotic exit network
(MEN), a signal transduction cascade that drives inactivation of mitotic cyclin-dependent kinases and is strictly
necessary for mitotic exit and cytokinesis. We are investigating the molecular mechanisms of this process.
3. REGULATION OF CYTOKINESIS BY Dma1/2 PROTEINS
We implicated two previously uncharacterized yeast proteins that we named Dma1 and Dma2 in the control of
cytokinesis. We showed that they are required, together
with the PAK kinase Cla4, for deposition of the septin ring
at the bud neck, which is in turn essential for proper spindle positioning and subsequent cytokinesis. In addition,
Dma1 and Dma2 participate to the spindle position checkpoint. Therefore, Dma1 and Dma2 are likely to be crucial
for preserving genome stability. Dma1/2 proteins are functionally redundant and they share the same structural
organization as S. pombe Dma1 and human Chfr and Rnf8,
which are all involved in checkpoint mechanisms. Dma1/2
proteins, as well as Chfr and Rnf8, are ubiquitin ligases
with a forkhead-associated domain that is normally implicated in the interaction with phosphorylated proteins, and
a Ring-finger domain typical of E3 ligases. We hypothesised that Dma1/2 may ubiquitinate protein(s) that regulate
septin ring assembly or function and we are trying to identify their possible targets through genetic screens and biochemical analysis of candidate proteins.
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5
SYSTEMS BIOLOGY
OF CELL PROLIFERATION
AND DIFFERENTIATION
PARTICIPANTS
Lilia Alberghina, Marco Vanoni, Paola Coccetti, Ferdinando Chiaradonna,
Anna Maria Colangelo, Elena Sacco, Claudia Cirulli, Valeria Mapelli,
Farida Tripodi, Daniela Gaglio, David Metalli. Daniele Colombo, Chiara
Balestrieri, Flavio Amara, Viktoria Tsiarentsyeva, Annalisa D’ Urzo.
The research groups of L. Alberghina and M. Vanoni are
developing a modular systems biology approach to the
study of cell cycle (most notably of the G1/S transition) in
the model organism, Saccharomyces cerevisiae, as well
as in normal and transformed mammalian cells. The
approach involves both “wet” experiments as well as
computer modelling and simulation. Experimental data
are used to extract information on network topology
leading to mathematical models and to estimate parameter
values. In order to understand this complex phenomenon,
it is mandatory not only to study the core machinery
driving the cell cycle, but also its modulation by genetic
and enviromental conditions, including nutrient and
growth factor availability, as well as the interconnections
with differentiation, signal transduction and cell death
pathways. These approaches should lead to a more rational
and more efficient drug discovery process.
CK2 CONTROL OF THE G1 TO S TRANSITION: NETWORK
IDENTIFICATION AND PARAMETER ESTIMATION
Paola Coccetti, Farida Tripodi, Claudia Cirulli, Marco Vanoni, Lilia Alberghina.
CK2 is a highly conserved enzyme ubiquitously distributed
among eukaryotes that phosphorylates a wide range of
substrates. Genetic studies indicate that CK2 activity is
required for cell cycle progression in both mammals and
yeast. In the latter organism, following CK2 depletion, cells
arrest cell cycle either in G1 and in G2/M. Recent results
newly indicate a major involvement of CK2 in the regulation of cell cycle progression in budding yeast since: (i) the
inhibitor of cyclin dependent kinase Sic1 is phosphorylated
in vitro and in vivo by CK2, (ii) Sic1 accumulates in a
cka1∆cka2ts strain and inhibits the Clb5-Cdk1 complex at
37°C, (iii) SIC1 deletion does not rescue the overall cell
growth defect of a cka1∆cka2ts strain, but almost completely abolishes the G1 block, (iv) the ubiquitin-conjugating
activity of Cdc34 involves CK2-mediated phosphorylation
of its catalytic domain. Based on our previous work on
CK2-mediated phosphorylation of Sic1 and Cdc34
(Coccetti et al 2006, Coccetti et al 2008), the goal of our
research is to elucidate the role of CK2 phosphorylation on
the G1/S transition in budding yeast studing by mass spectrometry the phosphorylation state of its relevant substrates (Sic1, Cdc34) as a function of growth conditions and cell
cycle position in exponential and perturbed growth.
NUTRITIONAL MODULATION OF CELL CYCLE PROGRESSION IN SACCHAROMYCES CEREVISIAE STUDIED BY
BIOCHEMICAL AND POST-GENOMIC TECHNIQUES
Marco Vanoni, Paola Coccetti, Stefano Busti, Valeria Mapelli, Farida
Tripodi, Viktoria Tsiarentsyeva, Lilia Alberghina.
Cell proliferation requires an exquisite coordination
between continuous events of the growth cycle and
discontinuous events of the nuclear division cycle which
results in cell mass homeostasis and correct duplication
and segregation of the genetic material.
In Saccharomyces cerevisiae, the mechanism that
controls entry into S phase requires overcoming of two
sequential thresholds –involving the cyclins Cln3 and Clb5
and their cognate inhibitors Far1 and Sic1 – that cooperate
in carbon source modulation of cell size. Mutants in glucose
transport, glucose sensing and physiological, biochemical
and post-genomic techniques are being used to study
nutritional modulation of the cell cycle both during exponential growth and during transient shifts from one carbon
source to the other with the final aim to obtain a qualiquantitative characterization of signalling pathways
affecting the G1/S transition and their connection to the
cell cycle machinery.
The relevance of two signalling pathways: TOR and
Snf1/AMPK on the G1 to S transition has been analysed:
TOR inhibition affects mainly Sic1 expression and stability
and Cyclins/Cdk activity (Zinzalla et al, 2007), while Snf1
deletion affects the translation, but not the transcription of
Clb5. Taking together all the previous results we are
moving towards a more complete network identification of
the G1 to S transition.
MODELLING OF THE G1/S TRANSITION IN YEAST
AND IN MAMMALIAN CELLS
Lilia Alberghina, Matteo Barberis, Ferdinando Chiaradonna, Daniela
Gaglio, Marco Vanoni.
We have previously proposed that the Cki Far1 cooperates
with Cln3 in a nutritionally modulated threshold controlling
Start execution and obtained experimental evidence for
the role of Far1 in the G1/S transition (Alberghina et al,
2004). These findings have been incorporated into a
mathematical model of the G1 to S network that newly
takes into account nucleo/cytoplasmic localization, the
role of Cki Sic1 in facilitating nuclear import of its cognate
1
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2
Cdc34
1_Mitochondrial morphology (green
staining) and cytoskeleton (red staining) NIH3T3 cells
2_A schematic representation of Cdc34 functional domains
Cdk1-Clb5, Whi5 control, and carbon source regulation of
Sic1 and Sic1-containing complexes (Rossi et al, 2005). In
collaboration with Edda Klipp (MPI, Berlin) the model was
implemented by a set of ordinary differential equations
that describe the temporal change of the concentration of
the involved proteins and protein complexes. The model
was tested by simulation in several genetic and nutritional set-ups and was found to be neatly consistent with
experimental data. A novel relevant conclusion comes
from our analysis: Ps is an emergent property of network
that strongly depends on growth rate (Barberis et al,
2007). A mathematical model of the entire cell cycle is
now under construction. Time-course analysis of key players in the G1/S transition of normal mammalian fibroblasts have been collected and are being integrated with
literature data to develop a model for the G1/S transition
of normal mammalian fibroblasts assuming a conservation during evolution of the basic structure of this network.
MECHANISMS OF NEURONAL APOPTOSIS AND OF
NEUROPROTECTION
Anna Maria Colangelo, Daniele Colombo, Flavio Amara, Lilia Alberghina.
Apoptosis is regarded as the main form of neuronal death
during neurodegenerative diseases. Global analysis of
neuronal apoptosis in Alzheimer Disease (AD) has led to a
modular molecular model where mitochondrial function
is modulated by molecules regulating survival/differentiation in response to Nerve Growth Factor (NGF)
(Alberghina & Colangelo, 2006). Given that neuronal
apoptosis is preceded by events of cell cycle re-entry, we
are using NGF-differentiated PC12 cells as in vitro model
to elucidate mechanisms linking mitochondrial dysfunction and cell cycle re-entry during neuronal apoptosis
following NGF deprivation and/or oxidative stress.
Besides, the different pathways of cell death and their
relation to longevity have been analyzed (Salvioli et al, 2008).
rhNGF AND NGF-LIKE PEPTIDES FOR THE THERAPY OF
NEUROPATHIES
Lilia Alberghina, Anna Maria Colangelo, Daniele Colombo, Enzo Martegani.
The role of NGF deprivation in several neuropathies, and
more specifically in Alzheimer Disease development and
progression is well established. Based on our previous
work on production of recombinant human (rhNGF)
(Colangelo et al., 2005), we are currently working in collaboration with two Companies, Primm and BluePrint
Biotech, on projects aiming at: i) bioassays for development of rhNGF; ii) development and in vitro and in vivo
analysis in comparison to rhNGF of NGF-like molecules
that might be characterized by better pharmacological
properties (Colangelo et al., 2008). An evolutionary study
of neurothrophins and of their receptors recently performed by our lab (La Nave et al, 2007) offers further information on their structure/function relation.
CANCER AND METABOLISM: ROLE OF ONCOGENIC
K-RAS PROTEIN IN METABOLIC REPROGRAMMING OF
CANCER CELL
Ferdinando Chiaradonna, Daniela Gaglio, Marco Vanoni, Lilia Alberghina.
The relation between alteration of metabolism and transformed phenotype have received increased attention
during the last few years. We study the mechanisms involved in such metabolic alterations with aim to elucidate the
role of the ras oncogene in cancer cell metabolic reprogramming. We showed that selective growth advantage
ras-transformed fibroblasts is lost upon growth in suboptimal glucose concentration (Chiaradonna et al., 2006).
Glutamine shortage, strongly reduces of the proliferation
ability of transformed cells, without inducing apoptosis and
with no effect on overall protein synthesis or ATP level. The
growth potential of transformed cells could be restored by
adding the four deoxiribonucleotides (but not Krebs cycle
intermediates), indicating that fragility of ras-transformed
cells to glutamine depletion is largely due to a reduced supply of DNA replication precursors in the presence of active
signalling inputs leading to execution of the G1/S transition.
DESIGN AND DEVELOPMENT OF RASGRF1-DERIVED
RAS INHIBITORS
Elena Sacco, David Metalli, Annalisa D’Urzo, Marco Vanoni.
Mutation of the ras genes is a critical event in the pathogenesis of different human tumors and in some developmental disorders termed “neuro-cardio-facial-cutaneous” (NCFC) syndromes.
Peptides derived from the Ras activator RasGRF1, showing both in vitro and in vivo Ras-sequestering properties
have been isolated (Sacco et al., 2005). RasGRF1-derived
peptides able to penetrate the plasma membrane by
means of their fusion with protein transduction domain
(PTD) of the Tat protein from HIV virus have been produced in collaboration with the Company Creabilis
Therapeutics and shown to be internalized by mouse
fibroblasts, when added to culture medium. The exogenously given peptides attenuate serum-mediated activation of the Ras-effector MAPK, reduce cell migration in
wound healing assays and reduce cell proliferation.
Further structure-guided down-sizing of the peptides is
underway.
REAL TIME ANALYSIS OF PROTEIN-PROTEIN INTERACTION
Marco Vanoni, Annalisa D’urzo, Elena Sacco.
The BIAcore technology is being used as an effective tool
to analyze protein/protein interaction and protein/ligand
interaction in real time. The technique is being applied
mostly to interaction of proteins of potential pharmaceutical interest, including the Ras oncoprotein, prion-derived peptides, cell cycle inhibitors and ataxin.
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1
6
SIGNAL TRANSDUCTION
IN EUKARYOTIC CELLS
1_UBPy in cortical neurons
(rat brain)
2_Localization of Cdc25-GFP
(green) in yeast nuclei.
DAPI (in red) stains DNA
PARTICIPANTS
Enzo Martegani, Sonia Colombo, Renata
Tisi, Michela Ceriani,
Fiorella Belotti, Chiara Paiardi, Cristina
Scandiuzzi, Loredana Amigoni
SIGNAL TRANSDUCTION IN YEAST:
Enzo Martegani, Sonia Colombo, Renata Tisi, Fiorella Belotti, Chiara Paiardi.
Collaboration with: Joris Winderixck and Rogelio Brandao
In Saccharomyces cerevisiae one of the main signalling
transduction pathways is the Ras/cAMP/adenylate cyclase
pathway, finely tuning pKA activity in the cell. The Ras-GEF
Cdc25 is essential for viability of yeast cells. Beside this
essential function related to its GEF activity, this protein is
revealing additional functions. Cell membrane fractionation
allowed to localize the Cdc25 protein in the internal membranes, but nuclei purification reveals that Cdc25 is also physiologically imported and efficiently retained in the nucleus.
Surprisingly, overexpression of the full length or of the
catalytical domain alone confers hyper-sensitivity to rapamycin, while the overexpression either of the N-terminal
region or of the 353-1100 aa region improves rapamycin
resistance, suggesting a cross-talk between Cdc25 and
Tor2 signalling. Cell membrane fractionation suggests that
also the Ras proteins localize on internal membranes and
not only on the plasma membrane. To study the localization
of active Ras2-GTP in vivo, we expressed the RBD of Raf1
fused with GFP. Our results show that active Ras appears
to be localised mainly inside the cells. As expected, upon
activation there is a rapid redistribution of the fluorescence
in the wild type strain. Adenylate cyclase activity is controlled by the Ras proteins and also by Gpa2, a heterotrimeric
G-protein. At present two Ras binding sites have been
mapped on adenylate cyclase (a RAD and a C-terminal
domain), while the regions of interactions with Gpa2 are not
yet known. Preliminary experiments suggest that the RAD
domain of Cyr1 is also involved in the interaction with Gpa2.
Another glucose-induced signal in Saccharomyces cerevisiae is a transient calcium increase mediated by phospholipase-C. The cytosolic calcium increase is mainly due to an
influx from external medium. Glucose-induced calcium
influx activates in its turn calcineurin, a calcium dependent
protein phosphatase involved in calcium homeostasis
regulation.
SIGNAL TRANSDUCTION IN MAMMALIAN CELLS:
Signal transduction mechanisms in NGF-mediated
differentiation.
Enzo Martegani, Michela Ceriani, Cristina Scandiuzzi, Loredana Amigoni
Collaboration with: Stefano Morara
The binding of NGF to TrkA activates the receptor and in
PC12 cells this promotes an activation of MAP kinases and
neuronal differentiation. Recently, the interaction with the
receptor and the phosphorylation of Ras Guanine
Releasing Factor 1 (RasGRF1) by TrkA have been shown in
Cos7 cells. RasGRF1 is a guanine nucleotide exchange factor for the Ras and Rho GTPases. Western blot analysis
demonstrated the TrkA-RasGRF1 interaction in PC12-TrkA
cells. PC12-TrkA cells, transfected with RasGRF1-GFP,
were also able to extend neurites even when they are not
stimulated with NGF. These observations bring us to identify RasGRF1 as a factor involved in the differentiation and
able to act synergistically with TrkA in the signalling
pathway. In addition we are interested to study the involvement of the deubiquitinating enzyme UBPy (USP8) in the
internalization and stability of the TrkA receptor. UBPy has
been found to interact with receptor tyrosine kinase like
EGFR and MET. This interaction is relevant for the recycling
of the receptor and modulates the signal transducion. We
found that mUBPy interacts with TrkA either in PC12 and in
Hek293 cells. This interaction reduced the level of ubiquitination of TrkA and therefore could stabilize the protein. In
addition we found that mUBPy is widely distributed in the
rat brain and is highly expressed in some brain areas that
also expressed TrkA, therefore we would like to verify the
functional role of the interaction between mUBPy and TrkA
in mammalian brain.
Small G-protein and APP processing:
Enzo Martegani, Michela Ceriani, Loredana Amigoni
Collaboration with: Tommaso Russo
The beta-amyloid precursor protein (APP) plays a central
role in the generation of Alzheimer’s disease. In this study
our attention was focused on identification of molecules
involved in the signal transduction mechanisms that affect
the enzymatic processing of APP. We used a system based
on HeLa cells expressing an APP-Gal4 fusion protein.
Factors able to affect the APP cleavage trigger also the cleavage of the fusion protein with release of Gal4 that activates
an UAS/Gal-luciferase reporter. We found that p21-hRas stimulated the APP cleavage, while no effect was observed with
other small G-protein (RalA, Rap1). A strong activation was
observed with an activated Ras (k-Ras Leu 61) and also
when Ras was cotransfected with an activated tyrosine kinase receptor (EGFR or TrkA).
2
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YEAST AS A MODEL
SYSTEM FOR STUDYING AGING AND
STRESS-RELATED PROCESSES
7
PARTICIPANTS
Marina Vai, Ivan Orlandi, Mario Dante, Matteo Viganò,
Ambra Corti, Pietro Giani, Gabriella Marincola, Eleonora Ottina,
Stefania Pessina, Marta Solinas, Daniele Stracka.
HISTONE MODIFICATIONS AND AGING IN
SACCHAROMYCES CEREVISIAE.
Marina Vai, Ivan Orlandi, Mario Dante, Ambra Corti, Pietro Giani, Gabriella
Marincola, Eleonora Ottina, Stefania Pessina, Marta Solinas, Daniele Stracka
DNA of eukaryotes is wrapped around nucleosomes
and packaged into chromatin. The details of this packaging are crucial for many cellular processes including
aging. Changes in chromatin are mediated by histones
modifications that include acetylation, methylation and
ubiquitination. The Sir2 family (Sirtuins), comprises the
unique class of NAD + -dependent deacetylases.
Sirtuins are phylogenetically conserved and beyond
silencing, they promote longevity. In yeast, proper
association of Sir2p to silent chromatin requires the
activity of a deubiquitinating enzyme, Ubp10 that regulates the levels of H2B-Lys123 ubiquitination. In addition, ubp10 cells display some features of aged cells.
We are focusing on the role of Ubp10 enzyme in the
regulation of the chromatin state, studying histones
modifications in some selected mutants in different
experimental conditions related to aging. Therefore,
processes such as glycolysis, respiration and NAD+
synthesis, that influence the pool of nicotinamide
metabolities, have a profound effect on Sirtuins activity. In this context, a molecular characterization of yeast
strains that have altered mitochondrial NADH/NAD+
ratios is underway (in collaboration with L. Palmieri,
Università di Bari, Italy).
logical role in fungal cells and its absence in mammalian ones. Since the integrity of the cell wall depends on
the synthesis and correct assembly of its components,
special emphasis has been directed towards a family of
beta (1-3)glucanosyltranferases that play an essential
role in fungal cell wall biogenesis. A functional characterization of these enzymes of Paracoccidioides brasiliensis is underway (in collaboration with C.M. de
Almeida Soares, Universidade Federal de Goiás,
Brazil). This fungus is the etiologic agent of one of the
most prevalent human systemic mycosis in Latin America.
RIBOSOME BIOGENESIS AND CELL SIZE CONTROL
Marina Vai, Matteo Viganò.
SFP1 encodes a zing-finger protein that promotes the
transcription of a large cluster of genes involved in
ribosome biogenesis in response to nutrients and
stress. Moreover, Sfp1p functions as a dose-dependent
cell size regulator at START. Chemostat cultivations
and Gene Chip analyses have been performed to assess
the effect of metabolism and carbon source on Sfp1p
activity (in collaboration with J.H. De Winde, Delft
University of Technology, The Netherlands). On-going
analyses in batch, aim to better define the alteration of
regulatory circuits observed by changing SFP1 dosage.
Particular attention is devoted to the PKA and TOR
pathways (in collaboration with L.Alberghina,
Università di Milano-Bicocca, Italy).
YEAST IN SPACE
THE FUNGAL CELL WALL AS A TARGET FOR ANTIFUNGAL
DRUGS Marina Vai, Ivan Orlandi.
Marina Vai, Ivan Orlandi.
Opportunistic fungal infections have increased dramatically in recent years as a result of increased immunosuppression associated with AIDS, organ transplants,
aggressive treatment of cancer and autoimmune disorders. Clinically important fungal pathogens display
varying degrees of tolerance to the widely used antifungals principally due to their lack of fungicidal activity.
The cell wall is regarded as a target for the development of new antifungal agents due to its essential bio-
On autumn, a suitable experimental equipment containing yeast living cells has been sent together with
astronauts in one of the space flights organized by ESA.
In this context, yeast has been used as a model system
for studying the effects of lack of gravity. On yeast cells,
returned from the space, experiments testing the activation of some pathways involved in the stress response have been performed (in collaboration with S.
Bradamante, C.N.R., Milano, Italy).
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8
PROTEIN
MASS SPECTROMETRY
PARTICIPANTS
Rita Grandori, Carlo Santambrogio,
Maria Samalikova, Elena Accardo.
Mass spectrometry (MS) is employed on one side as an
analytical tool for proteomics. The focus is on the phosphorylation states and intracellular interactions of regulatory proteins involved in the control of the yeast cell
cycle. On the other side, mass spectrometry is applied to
the direct investigation of non-covalent interactions and
intact protein structures for conformational studies and
binding analysis. This information is complemented by
data obtained by other spectroscopic methods, such as
infrared spectroscopy (IR) and circular dichroism (CD).
SIC1 PHOSPHORYLATION
Rita Grandori, Maria Samalikova.
The cyclin-dependent protein kinase inhibitor Sic1 is
the yeast homologue of mammalian p21Cip1 and
p27Kip1 and is the key regulator of cell cycle progression and its interdependence with cell growth. It is
phosphorylated in vitro and in vivo by the CK2 protein
kinase, but the physiological role of such a modification is not understood yet. Analysis of Sic1 phosphorylation inside the cell is hindered by the low protein
concentration and the transient and labile nature of
the modification. We have developed a robust immunoaffinity/mass spectrometry (MS) approach for Sic1
phosphopeptide analysis in integrative transformant
yeast cells. The method has been employed to investigate the influence of carbon source on Sic1 phosphorylation by CK2. The results indicate that Sic1 phosphorylation is modulated by the cell-cycle phase and
by the carbon source. These findings allow us to formulate hypotheses about the molecular mechanisms
that mediate Sic1 role in the G1-to-S transition of the
yeast cell cycle. In collaboration with L.Albeghina and
M.Vanoni, this Department.
BETA-LACTOGLOBULIN FOLDING INTERMEDIATES AND
DIMERIZATION
Rita Grandori, Gaetano Invernizzi, Carlo Santambrogio.
Partially folded forms are of big relevance for the understanding of the mechanism of protein folding.
Trifluoroethanol (TFE) at low concentrations induces formation of a beta-lactoglobulin (BLG) intermediate that
contains non-native helical structure and that is thought
to form also under physiological conditions, playing a
role in BLG folding in vivo by preventing aggregation.
Such an intermediate could be detected by nano-ESI-MS
spectra with maximum accumulation at 16% TFE, in
excellent agreement with results from solution experiments. The synergism of TFE with generic denaturing
agents in promoting formation of the BLG intermediate
could be analyzed.
The BLG monomer-dimer equilibrium was analyzed by
nano-ESI-MS at variable protein concentration, pH and
interface temperature. The apparent dissociation constant and its pH dependence are in good agreement with
solution data. The effect of temperature provides apparent deltaH values that will be compared to calorimetry
data under identical solvent conditions.
PROTEIN-LIGAND INTERACTIONS IN THE TETRAMERIC
FLAVODOXIN-LIKE PROTEIN WRBA
Rita Grandori
WrbA represents a conserved family of proteins involved
in cellular stress response that bridges structurally and
functionally flavodoxins to quinone oxidoreductases.
Combined MS, CD and IR data indicate that FMN (flavin
mononucleotide) binding promotes WrbA oligomerization
resulting in increased protein thermostability. This study
illustrates the specific contribution that each technique
can give to the analysis of linked binding equilibria, particularly when the complexity of the system is magnified,
like in this case, by a high number of different species
involved and by irreversible unfolding due to protein
aggregation. In collaboration with S.M. Doglia, this
Department and Jannette Carey, Princeton University, NJ.
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PROTEIN ENGINEERING AND
INDUSTRIAL ENZYMOLOGY
9
PARTICIPANTS
Marina Lotti, Stefania Brocca, Pietro GattiLafranconi, Gaetano Invernizzi, Serena Caldarazzo,
Vera Codazzi, Marina Francesca Thurner.
Enzymes employed in biocatalysis, in particular lipases, and model proteins such as lactoglobulin are studied by a combined approach of mutagenesis (both
directed evolution and site directed mutagenesis) and
biochemical and biophysical characterization, in order
to highlight the molecular bases of their stability, function and propensity to aggregate upon expression in
bacterial cells. Cold adapted enzymes are used as
models to understand the structural determinants of
activity at low temperature. Moreover, novel biocatalysts are isolated from non commercial sources or
produced by protein engineering.
SOLUBILITY AND AGGREGATION OF RECOMBINANT
PROTEINS
The fine detail of the aggregation of recombinant proteins upon overexpression in E. coli cells is investigated
using several proteins and mutants thereof. The kinetics
of inclusion bodies formation as well as the residual activity and native-like structure of aggregated proteins is studied by biochemical and biophysical means. We observed
that the physiology of expression affects both the partition
of the protein between the soluble and aggregated fraction
and also the conformational state of the aggregated proteins. This work is performed in collaboration with S. M.
Doglia (Biophysics, our Department) and A. De Marco
(IFOM, Milano).
CONFORMATIONAL STABILITY OF PROTEINS
The stability of proteins to different experimental and/or
physiological conditions strongly impacts on their functionality and propensity to aggregation. We are studying different model systems to understand how function and
conformation are related by a combined approach of biochemical assays and mutagenesis strategies as well as by
biophysical techniques enclosing Fourier Transform infrared spectroscopy and nano-electrospray-ionization mass
spectrometry (nano ESI-MS) performed in partner laboratories. Proteins employed in these studies are beta-lactoglobulins, bacterial lipases and proteins involved in the cell
cycle. The effect of protein sequence as well as of posttranslational modifications (i.e. glycosylation) is also investigated. This work is performed in collaboration with S.M.
Doglia, R.Grandori and L.Alberghina from this Department.
MOLECULAR BASES OF SPECIFICITY, STABILITY AND
COLD-ACTIVITY IN LIPOLYTIC ENZYMES
The aim of this research is to gain insight in the molecular
determinants of specificity and stability in enzymes of interest in biocatalysis. We are investigating in depth the lipase produced by the bacterium Pseudomonas fragi (PFL).
Our interest in this protein relies on two aspects: i) it is a
cold-active, extremely thermosensitive enzyme, ii) it
displays a marked and unusual preference toward substrates containing short-chain fatty acids. A combined
approach of directed evolution and domain swapping with
regions of more stable homologous lipases is applied to
understand the molecular mechanisms at the basis of
adaptation to low temperture. In this project, a biocatalyst
stable enough to be employed in biotransformations and
yet retaining cold-activity was obtained.
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10
STRUCTURE
FUNCTION-PATHOGENICITY
RELATIONSHIPS
IN PROTEINS
PARTICIPANTS
Paolo Tortora, Maria Elena Regonesi, Emanuela Occhipinti, Marta Del Favero,
Matteo Riva, Elena Galbusera, Elisa Mazzantini, Serena Mazzucchelli.
Major topics in protein chemistry are the understanding of
the structure-function relationship and of the mechanisms by which some proteins are capable of triggering
specific diseases. We address these issues by studying
structural and functional properties of the proteins under
investigation, as well as their intracellular localization
and interactors. As far as enzyme proteins are concerned,
their catalytic behavior and sensitivity to inhibitors and activators are characterized. Structural features, in particular
the aggregation state, are explored by FT-infrared spectroscopy and atomic force microscopy. Intracellular interactors are identified by advanced mass spectrometry techniques. These approaches are matched with the development and characterization of mutated forms of the proteins under investigation, which helps clarify the structural properties associated with function and pathogenicity.
STRUCTURAL STUDIES ON PROTEINS CONTAINING
GLUTAMINE REPEATS RESPONSIBLE FOR NEURODEGENERATIVE DISORDERS
Maria Elena Regonesi, Paolo Tortora, Matteo Riva, Elena Vismara.
Some neurodegenerative disorders result from the
expansion of glutamine repeats (poly-Q diseases) in a set
of proteins. Their misfolding and aggregation are likely to
be involved in these disorders. The aim of this investigation is to gain insight into the molecular mechanism(s) by
which expanded poly-Q stretches in ataxin-3 lead to the
Machado-Joseph neurodegenerative disease. We are
focusing on two major issues related to the molecular
mechanism of the pathogenesis, i.e., the understanding of
the protein’s physiological role, and the mechanisms by
which ataxin-3 generates amyloid fibrils.
ROLE OF POLYNUCLEOTIDE PHOSPHORYLASE IN
MATURATION OF PROKARYOTIC TRANSCRIPTS
Paolo Tortora, Marta Del Favero, Elisa Mazzantini.
Polynucleotide phosphorylase (PNPase) is a prokaryotic
enzyme that degrades RNAs phosphorolytically. It plays a
major role in regulation of their stability, degradation and
maturation. This project is aimed at providing a better
Amyloid fibrils generated by human
ataxin-3, as shown by atomic force
microscopy. The arrows highlight
regularly spaced ridges.
insight into the role of PNPase in the aforementioned
degradative mechanisms and the factors which control its
activity. To this end, we take advantage of a set of mutants,
which are being characterized in terms of physiological
behavior, enzymatic properties and aggregation state.
INVESTIGATIONS ON STRUCTURE, STABILITY AND
FUNCTIONS OF PROTEINS FROM THE ARCHAEON
SULFOLOBUS SOLFATARICUS
Paolo Tortora, Emanuela Occhipinti.
S. solfataricus carboxypeptidase (CPSso) is a thermostable
metalloenzyme endowed with broad substrate specificity
and the ability to withstand extreme chemical-physical
conditions, such as temperatures up to 85°C and high
concentrations of organic solvent. A process aimed at
synthesizing N-blocked amino acids in organic medium is
being developed by taking advantage of the properties of
CPSso. Also, by combining mass spectrometry, molecular
modeling, and site-directed mutagenesis we could identify
key structural features responsible for its thermostability.
STUDIES ON THE PHYSIOLOGICAL ROLES OF HUMAN
CGI-58 PROTEIN
Paolo Tortora, Emanuela Occhipinti, Serena Mazzucchelli.
The intracellular accumulation of triglycerides is finely
regulated in adipocytes at the level of the so-called “lipid
droplets”, with which several proteins are associated.
Such organelles play a role in triglyceride storage,
transport and degradation. CGI-58, a lipid dropletassociated protein, is of unknown function. However,
individuals suffering from the Chanarin-Dorfman syndrome, a rare hereditary disorder resulting in lipid accumulation, carry different mutations in the CGI-58-encoding
gene. This points to a major role for this protein in lipid metabolism. Our investigation is aimed at elucidating the physiological role(s) of CGI-58, and the molecular mechanisms of the aforementioned disease.
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STRUCTURAL AND
FUNCTIONAL STUDIES
ON PROTEINS
11
PARTICIPANTS
Alessandra Bigi, Lavinia Morosi, Valentina Pastori, Alessio
Somaschini, Chiara Pozzi and Paola Fusi.
STUDIES ON ATAXIN-3 PHYSIOLOGICAL ROLE
Valentina Pastori, Alessio Somaschini, Chiara Pozzi and Paola Fusi.
Spinocerebellar ataxia type 3 (Sca3) is a poly-Q disease,
caused by the presence of an expanded polyQ stretch inside the coding sequence of ataxin-3, which leads to the formation of amyloid fibrils. Ataxin-3 physiological function is
still debated, although a role in ubiquitin mediated protein
degradation is widely accepted. In our laboratory we have
been carrying out a subcellular localization study, in transfected cells, using ataxins-3 with different polyQ lengths.
Results showed a mainly cytosolic localization for both
pathological and non pathological ataxins-3, but also showed that ataxin-3 is found in mitochondria. Toxic effects of
pathological ataxin-3 on mitochondria are currently investigated. Our results also showed that ataxin-3 is extensively proteolyzed, while the pathological form is more resistant to proteolysis. Edman degradation and mass spectrometry analysis of fragments allowed to identify cleavage sites, in collaboration with Tedeschi (University of
Milan). The role of Ataxin-3 phosphorylation by casein
kinase 2 (CK2) is also investigated in collaboration with
P.Coccetti (This Department).
CHARACTERIZATION OF HUMAN SIALIDASES
Alessandra Bigi, Lavinia Morosi, Chiara Pozzi and Paola Fusi
Sialidases or neuraminidases are widely distributed glycohydrolytic enzymes removing sialic acid residues from glycoproteins and glycolipids. In mammals, several sialidases
have been described: a lysosomal form (NEU1), a soluble
enzyme (NEU2) and two membrane-associated forms
(NEU3 and NEU4). The structure of the soluble human sialidase NEU2 was elucidated by our group in collaboration
with S. Wakatsuki (Head of KEK Structural Biology Group,
Tzukuba, Ibaraki, Japan). A detailed kinetic characteriza-
tion showed that this enzyme has a broad range of action,
as well as a specificity towards substrate supramolecular
organization. Mutants have been produced to validate
NEU2 crystallographic structure and verify the proposed
catalytic mechanism. More recently, molecular dynamic
studies have been undertaken, in collaboration with L. De
Gioia and G. Zampella (This Department), to elucidate binding to ancillary substrate site, with the aim of designing
inhibitors more selective towards viral sialidases than
human ones, to be used as antiviral agents.
Characterization of membrane bound sialidase NEU4 is
also studied. Solubilization studies showed that NEU4 is
an extrinsic membrane protein, anchored to the membrane though interaction with other protein(s). Cross-linking
studies are currently carried our to identify these proteins.
Moreover, NEU4 membrane anchoring mechanism is
investigated, through site-directed mutagenesis.
STUDY OF THE MECHANISM OF CROSS-PRESENTATION
OF TUMOR ANTIGENS FROM BACTERIA-INFECTED
MELANOMA CELLS
Chiara Pozzi and Paola Fusi.
In M Rescigno’s laboratory, at the European Institute of
Oncology (IEO) in Milan, a new immunotherapy protocol for
metastatic melanoma patients has been developed, based
on the vaccination of patients against Salmonella followed
by the intratumoral injection of a non-virulent, but invasive, strain of S. typhimurium. We collaborate with this
group in the frame of a study aimed at understanding the
basis of the observed systemic anti-tumor response and at
elucidating the bacterial determinants responsible for this
phenomenon. Results suggest that bacteria facilitate processing of tumor antigens within the tumor cell and that
these antigens are transferred to the dendritic cells (DCs)
via gap junctions without the need of phagocytosis.
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wavenumber (cm-1)
12
1_WrbA thermal unfolding by FTIR spectroscopy
MOLECULAR
AND CELLULAR
BIOPHYSICS
second derivatives
1
PARTICIPANTS
Silvia Maria Doglia, Antonino Natalello,
Anna Maria Villa.
2_Confocal fluorescence image of
mitochondria in breast carcinoma
cells after EB (red)-R123 (green)
staining
PROTEIN SECONDARY STRUCTURE, STABILITY AND
AGGREGATION
S.M.Doglia, A.Natalello.
Protein stability and aggregation, central issues in biotechnology and medicine, have been studied in vitro and in
vivo on several model systems through complementary
biophysical and biochemical methods.
In particular, the effect of several chemical and physical
effectors, as well as of the osmolyte betaine, on protein
misfolding and aggregation has been investigated in vitro,
in collaboration with the research group of Dr.A. de Marco
(IFOM, Milan).
In addition, the role of Flavin mononucleotide on the
stability and oligomerization of the tryptophan repressor
binding protein A (WrbA) has been studied in collaboration
with Prof. R. Grandori of this Department by Fourier
transform infrared (FT-IR) spectroscopy.
Furthermore, in collaboration with the group of Dr M.
Salmona (Istituto di Ricerche Farmacologiche “Mario
Negri”, Milan) we characterised the kinetics of aggregation
of the human prion peptide PrP82-146 and the structural
properties of its oligomers and fibrils by FT-IR spectroscopy,
electron microscopy and confocal fluorescence microscopy.
High resolution atomic force microscopy was also performed
in collaboration with V. Prokhorov (Institute of Bioorganic
Chemistry, Russian Academy of Sciences, Moscow).
Preliminary results indicated that the PrP82-146 peptide
was capable of undergoing several aggregation pathways,
with end products displaying different secondary structures and intermolecular interactions. These findings point
to a high plasticity of the prion peptide, a crucial feature of
prion proteins to overcome species barriers.
Concerning the study of protein aggregation in vivo, we
recently proposed a new FT-IR method (Ami et al. FEBS
Lett. 2005) to monitor the aggregation of recombinant
proteins in bacterial cells in the form of inclusion bodies
(IB). This approach enabled us, in collaboration with the
group of Prof. M. Lotti of this Department, to follow the
2
kinetics of IB formation in a rapid way and to obtain
structural information on proteins within IBs. The analysis
of protein FT-IR spectra indicates that a residual nativelike structure is retained to a varying extent within IB,
depending on the level of the recombinant protein expression, with interesting biotechnological implications. The
disaggregation of IBs and the role of the chaperone DnaK
has been also studied in collaboration with the research
group of Prof. A. Villaverde (Autonomous University of
Barcelona, E).
CONFOCAL FLUORESCENCE MICROSCOPY AND FT-IR
MICROSPECTROSCOPY OF INTACT CELLS.
S.M.Doglia, A.M.Villa, A.Natalello.
Recently, by fluorescence confocal microscopy of
Rhodamine 123 (R123) we detected - at single cell level - the
presence of two populations of mitochondria in carcinoma
cells, differing for their localization, morphology and
membrane potential. Further studies of intracellular
distribution of ethidium bromide (EB) in the same cell
lines, unexpectedly, indicated that EB cannot be taken as
an overall marker of mitochondria. Indeed, in breast and
in lung carcinoma cells, a different EB fluorescence
intensity was observed for the two populations. We,
therefore, investigated the possible correlation between
EB fluorescence and the accessibility of mtDNA in active
transcription and replication. To this goal, in collaboration
with the group of Dr P. Fusi of this Department, we studied
mitochondria of neuroblastoma cells, where mtDNA
replication was modulated by differentiation. Preliminary
results indicated a correlation between the intensity of EB
fluorescence and the percentage of mtDNA nascent
strands.
In collaboration with the group of Prof. C.A. Redi
(University of Pavia, Italy), we studied the differentiation of
murine embryonic stem cells in their early development
by FT-IR microspectroscopy. As supported by cytochemical
essays and by the statistical analysis of the spectral data,
the FT-IR approach enabled us to monitor in a rapid way
the temporal evolution of the ES cell differentiation (Ami
et al. BBA-MCR 2008).
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THERAPEUTICAL
STRATEGIES
FOR CHRONIC PAIN
13
PARTICIPANTS
Gabriella Giagnoni, Barbara Simona Costa,
Francesca Comelli, Isabella Bettoni
EFFECT OF THE MODULATION OF THE ENDOCANNABINOID
SYSTEM ON INFLAMMATORY AND NEUROPATHIC PAIN.
In the past, the Cannabis sativa plant was used for both
recreational and medicinal purposes. Only in 1990s the
interest in natural and synthetic cannabinoids was reconsidered with the discovery of cannabinoid receptors and
their endogenous ligands such as anandamide and 2-arachidonoylglycerol. These endocannabinoids are synthesized and released on demand in response to pathological
and physiological stimuli, such as inflammation and pain.
On these bases, we studied whether the modulation of the
endocannabinoid system results in relief of pathological
pain. Particularly we showed that: a) both the non-psychoactive Cannabis component, cannabidiol, and a Cannabis
extract enriched in cannabidiol, have a potent analgesic
effect in two models of neuropathic and inflammatory
pain; b) the employment of selective compounds able to
inhibit the endocannabinoid reuptake (AM404) or intracellular degradation (URB597 and URB602), was an innovative and efficacious tool to alleviate neuropathic pain avoiding the psychoactive effects normally associated with
exogenous administration of cannabic agonists; c) a stable
structural hybrid between an endocannabinoid and an
endovanilloid, like arvanil, evoked a significant decrease of
chronic inflammatory pain, so suggesting that the simultaneous modulation of both endogenous systems is a valid
approach to treat this painful condition.
MICROGLIA AS A NEW TARGET FOR THE TREATMENT OF
NEUROPATHIC PAIN
Recent evidence indicated that microglial cells in the spinal cord are critically involved in the development and
maintenance of neuropathic pain, with a pivotal role of
TLR4 receptor. Binding of an endogenous ligand to TLR4
might be considered an important step in the regulation of
microglia activity in pain facilitation, suggesting that a
mechanism aimed to inhibit such a binding could be
potentially effective against neuropathic pain. Prof. Peri of
our Department recently synthesized new ligands to TLR4
with antagonistic activity. In the present work we evaluated
the efficacy of the most potent TLR4 antagonist synthesized by us (FP-1), administered in mice with painful neuropathy. The repeated treatment of neuropathic mice with
FP-1 evoked a relief of both thermal hyperalgesia and
mechanical allodynia, whereas its administration to TLR4
knockout neuropathic mice revealed that in the absence of
TLR4 receptor, the compound lost its efficacy. As consequence of TLR4 binding, the repeated treatment with FP-1
prevented the activation of the transcription factor NF-kB
and the TNFalpha overproduction in the spinal cord.
Together, our findings support the previous evidence indicative for a contribution of glial TLR4 to the initiation of
neuropathic pain, suggest it as potential innovative target
to treat this debilitating disease, and propose FP-1 as lead
compound for the development of new effective drugs.
MESENCHYMAL STEM CELL TRANSPLANTATION AS A
THERAPEUTIC APPROACH FOR NEUROPATHIC PAIN
Neuronal transplantation is one of the most innovative
therapeutic potential approach for the treatment of peripheral neuropathies or degenerative pathologies.
Recently, also stromal stem cells derived from the bone
marrow (MSCs) could be use for neuronal transplantation,
because they are easily collected from bone marrow, rapidly expanded in culture and, under specific conditions, they
can differentiate, both in vitro and in vivo, in neuronal cells,
in astrocytes or in cells able to product myelin, thus facilitating nerve regeneration and remyelination. Thus we
decided to, transplant MSCs in the spinal cord of mice with
chronic constriction injury of the sciatic nerve. This transplantation partially reduced thermal hyperalgesia and
mechanical allodynia, already 7 days after the nerve injury.
In order to improve the antinociceptive effect of the stem
cell transplantation, a major number of cells were injected
to neuropathic mice. This treatment ameliorated thermal
hyperalgesia without improving mechanical allodynia. The
employed model of neuropathic pain is characterized by
morphological modifications of Schwann cells, axonal
degeneration, myelin loss, all critical events responsible
for pain. Since MSCs are able to promote nervous tissue
reparation releasing cytokines and neurotrophic factors,
we further investigated whether the stem cell transplantation in neuropathic mice promote nerve regeneration and
remyelination. The histological analysis performed on
sciatic nerve portion proximal to nerve ligation showed
that MSCs transplantation favoured the remyelinization
process. In conclusion, the data clearly suggested that the
cellular therapy with MSCs could be an innovative approach for the management of peripheral neuropathies.
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REGULATION OF NEURAL STEM
CELLS IN PHYSIOLOGY AND EXPERIMENTAL
THERAPY FOR CANCER AND
NEURODEGENERATIVE DISORDERS.
14
PARTICIPANTS
Angelo L.Vescovi, Serena Acquati, Elena Binda,
Roberta Fiocco, Fabrizio Gelain, Daniela Ferrari,
Lidia De Filippis, Silvia Panseri, Sara Piccirillo,
Valeria Rigamonti, Laura Rota Nodari, Joyce
Nwachukwu, Francesca Taraballi, Natasa Zarovni,
A
B
Engrafted IhNSC into the corpus callosum.
(A) and hippocampal dentate gyrus (B) of ischemic adult rat brains at 1
month from transplantation (magnification 40x).
NERVOUS REGENERATION VIA NANO-STRUCTURED
SCAFFOLDS
Fabrizio Gelain, Francesca Taraballi, Silvia Panseri, Valeria Rigamonti,
Angelo L.Vescovi.
A traumatic injury to adult nervous system often leads
to persistent deficits, due to the inability of mature
axons to regenerate after damage, which results on a
significant impact on quality of life and life expectancy
for the patients. Our project focuses on traumatic injury both in central and peripheral nervous system. In
order to enhance nervous regeneration our approach
make joint use of diverse nanotechnology derived biomaterials: electrospun micro- and nanofiber channels
and self-assembling peptides. Both are bio-reabsorbable and have been shown not to elicit marked immune response, nor inflammatory reactions in animals.
Electrospun tubes are flexible tubular scaffolds showing high porosity and surface/volume relation. Selfassembling peptides are made from natural amino
acids, they undergo self-assembly into nanofibers forming a scaffold, they can be mixed with growth factors
before the self-assembling takes place upon exposure
to neutral pH solutions. In the last year we demonstrated how electrospun tubes can be used to successfully
regenerate a 10-mm nerve gap of transacted rat sciatic nerve in vivo. Our work provided evidence that electrospun micro- and nanofiber channels are promising
bioabsorbable scaffolds for stimulating and guiding
peripheral nerve regeneration in rat models of sciatic
nerve transection. Our approach is going to be further
ameliorated via complementary strategies like hydro-
gels for drug delivery, cell transpantation and techniques adopted in clinics like physiotherapy. Other experiments are assessing the regenerative potential of a
similar nanotechnology based methodology in animal
models of contusive spinal cord injuries.
TRANSPLANTATION OF ADULT NEURAL STEM CELLS
(HNSC) FOR TISSUE REPAIR AND STEM CELL THERAPY: CHARACTERIZATION OF HNSCS IMMORTALIZED
WITH v-myc (ihNSC) IN VITRO AND IN VIVO
Lidia De Filippis, Daniela Ferrari, Laura Rota Nodari, Angelo L.Vescovi.
Neural stem cells represent a source of fundamental
relevance for the therapeutic approach to neurodegenerative diseases like Parkinson’s or multiple sclerosis. A cell line of human neural stem cells already
available in our lab (IhNSC) was immortalized with a
retroviral vector carrying the v-myc gene. IhNSCs were
characterized in vitro by proliferation and differentiation assays showing they retain basal stem cell properties like self-renewal and multipotency. In particular, their proliferative potential was 4-5 fold increased
with respect to parental cells in the presence of the
growth factors EGF and FGF2; upon removal of mitogens, IhNSC spontaneously generate astrocytes and
significative quantities of neurons and oligodendrocytes able to survive up to 40 days in vitro. To validate
IhNSC as a non tumorigenic cell line, we transplanted
IhNSC into the brain of nude SCID mice and after 6
months from transplantation no tumor developed. In
order to characterize the potentialities for therapeutic
purposes of IhNSC in vivo, IhNSCs were transplanted
into the ischemic brain of adult rats. By immunofluo-
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rescence assay, IhNSC were detected after 1 month
from transplantation in the corpus callosum and cortex and dentate gyrus of hippocampus, as both astroglial GFAP-IR cells, and neuronal cells labeled with
the ß-tubulinIII and the MAP2 (dendritic) markers.
Current studies are aimed to assess a functional integration of human cells as mature neurons.
ISOLATION AND FUNCTIONAL CHARACTERIZATION
OF BRAIN TUMOUR STEM CELLS DERIVED FROM
HUMAN ADULT GLIOBLASTOMAS
Sara Piccirillo, Elena Binda, Joyce Nwachukwu, Angelo L.Vescovi.
Gliomas are the most common brain tumours, and, in
particular, grade IV glioma, named glioblastoma multiforme (GBM), represents the most aggressive type. Due to
the highly diffuse pattern of this heterogeneous tumour,
prognosis is severe and life expectancy generally ranges
from 17 to 50 weeks. In essence they are incurable.
In the last years, transformed stem cells have been isolated from some non-solid and solid human cancer. We
reported that, unlike other brain tumours, the lethal
GBM contains neural precursors endowed with all the
critical features expected from neural stem cells. Most
importantly, these cells are tumorigenic, in fact they are
able to reproduce the original neoplasia upon injection
into the brain of immunosuppressed mice.
Starting from these results, we have collected GBM
specimens from patiens undergoing neurosurgical
resection for high grade glioma and from these we
have established stable tumour neural stem cells lines
(TNSCs) that have been characterized at the cellular,
chromosomal and molecular levels, particularly with
regard to their stem cells properties and expression of
novel candidate markers, with an immunophenotypic
analysis of approximately forty markers.
We also developed in vitro high throughput drug screening assays, to identify panels of chemotherapy drugs
to which TNSCs may be more susceptible. The most
efficacious treatment emerging from the in vitro
screenings shall then be used to try to prevent or cure
tumours in our GBM animal model. This will allow to
determine the actual degree of reliability of these
assays in predicting the drugs efficacy in slowing or
blocking tumour development.
PLURIPOTENCY ASSOCIATED GENES IN NSCs (PLURIGENES)
Angelo L. Vescovi, Natasa Zarovni, Serena Acquati, Roberta Fiocco, Joyce
Nwachukwu.
Understanding the mechanisms controlling multi/pluripotency in adult NSCs and ES holds promise of
extending the potential and plasticity of neural cells
and thus develop new sources and strategies for regenerative therapies. A systematic analysis of available
experimental data on gene expression during NSCs
differentiation defined the list of interesting candida-
tes candidate genes potentially involved in regulation
of self renewal and fate choice in adult NSCs in vitro
and in vivo.
Recent findings show that certain adult progenitors as
well as adult somatic cells remain competent to interpret key signals that are responsible for maintenance
of pluripotency in ES. In view of these results a clustering analysis between expression patterns in adult
murine NSC and ES could enable us to identify stemness factors conserved in different stem cell populations. Genes of interest are expressed in neurogenic
areas of adult mouse brain. Moreover, for a subset of
selected genes a “pathological” activation has been
reported in tumor cell lines and some human cancers.
In order to study their role in the maintenance of the
stem cell functional determinants and cell plasticity,
knock down and over-expression experiments are
ongoing using both tools for constitutive and tetracycline regulated systems for shRNA and cDNAs expression. Stable murine and human NSC clones have been
selected for evaluation of kinetics of expansion, cell
viability and differentiation profile.
INDUCTION OF OLIGODENDROCYTES FROM NSCs AND
ES-DERIVED NEURAL PRECURSORS
Angelo L.Vescovi, Serena Acquati, Natasa Zarovni.
Several studies have demonstrated that oligodendrocyte precursors (OPCs) can promote remyelination and
regeneration after being transplanted in injured animals. OPCs, are lineage-restricted progenitors deriving from multipotent NSCs that can yield mature oligodendrocytes during adult life as a response to environmental stimuli. Oligodendrocyte development both
in vivo and in vitro is a good example of how the identity of a distinct cell type is affected by extracellular
factors and the specific signalling pathways but also of
how the cellular memory is important for responding
to extracellular cues.
We optimized protocols for OL induction from adult
murine NSCs as well as ES derived NSC- like lines.
Stable ES and NSC cell lines have been generated in
which GFP expression is driven by the CNP promoter
activity. Thus, premyelinating and proliferating OPCs
can be selected, purified and amplified for transplant
studies. Final milestone of the project is to test OPCs
for their therapeutic potential in a mouse model of MS.
The yield of oligodendrocytes from murine ES-derived
NSC like cells is found to be dependent on the neuralization protocol used. In the adult NSCs the oligodendrocyte fate was favored upon: treatment with purine
nucleotides/nucleosides, long term culture and growth
factor modulation. In addition, in some brain tumors a
percentage of oligodendrocytes is increased with
respect to “normal” proportions.
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1
15
DENDRITIC CELLS IN
INNATE AND ADAPTIVE
IMMUNITY
PARTICIPANTS
Paola Castagnoli, Francesca Granucci, Maria Foti, Ivan Zanoni,
Matteo Urbano, Federica Mainoldi, Anna Ranghetti, Anna Torri,
Silvia Fumagalli, Francesca Pontiroli, Roberto Spreafico,
Caterina Vitali, Caterina Bodio, Renato Ostuni, Simona Barresi.
2
1_Dendritic cells
and Bacteria Interactions
2_Dendritic cell-T
cell crosstalk.
Dendritic cells (DC) are a special type of leukocytes able to
alert the immune system for the presence of infections.
They are extremely versatile antigen presenting cells
involved in the initiation of both innate and adaptive immunity, but also in the differentiation of regulatory T cells
required for the maintenance of self-tolerance. Multiple
animal models of infections and autoimmunity are used to
investigate how DC can mediate all these diverse and
almost contradictory functions.
DENDRITIC CELLS BIOLOGY AND MOLECULAR MEDICINE
Development of innate and adaptive immune response
during the course of a microbial infection is dependent
upon early interactions between incoming microrganisms with immature dendritic cells (iDCs) which are the
first immune cells interacting with the microbial agents.
The recent improvements of sequencing technologies,
and in particular the publication of the initial version of
the human and mouse genome sequences, have opened
the field of large-scale functional approaches of biological systems. We employ high-throughput technologies to
investigate fundamental aspects of the immune system
and their roles in health and disease. In order to identify
key cellular genes involved in these processes, we use a
transcriptomic approach in which modifications of cellular transcriptome are analysed at several times postinfection.
DENDRITIC CELLS AND NATURAL KILLER CELLS
Natural Killer (NK) cells exert a direct anti-tumor and antimicrobial effect and can influence the development of
adaptive T cell responses. Activation of NK cells is regulated by accessory cells such as dendritic cells (DC).
Following activation, NK cells accumulate at the lymph
nodes draining the site of infection, the key place in which
DC and NK cell interactions occur. Taking advantage of the
two-photon intravital microscopy technology the capacity
of activated NK cells to reach the draining lymph nodes is
investigated together with the DC-derived signals necessary for NK cell priming in inflammatory conditions induced by lipopolysaccharides.
DENDRITIC CELLS AND REGULATION OF IMMUNE
TOLERANCE
The immune system of vertebrate animals has the capacity to respond to perturbations (invading pathogens, stress
signals) limiting self-tissue damage. Tolerance to tissue
antigens is achieved through a combination of thymic and
peripheral events that eliminate or inactivate potentially
dangerous T cells. Several mechanisms have been proposed to explain the induction of tolerance in peripheral
autoreactive T cells. Taking advantage of different transgenic and knock out mouse models the mechanisms through
which dendritic cells induce T cell tolerance in peripheral
lymphoid organs are investigated.
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NEUROPHYSIOLOGY
16
PARTICIPANTS
Francesca Gullo, Enzo Wanke.
FUNCTIONAL STUDIES ON NA+ CHANNEL MUTATIONS
SPATIOTEMPORAL EVOLUTION OF NEURONAL
IN FEBBRILE EPILEPSY AND GENERALIZED EPILEPSY
NETWORKS INVESTIGATED WITH MULTIELECTRODE
WITH FEBBRILE SEIZURE (GEFS+)
ARRAYS (MEA)
Enzo Wanke.
Francesca Gullo, Enzo Wanke.
Febrile seizures (FS) affect 5-12 % of infants up to six
With the acquisition of a novel multielectrode array
years of age. Familial epilepsies are often caused by
(MEA) electrophysiological system we aim at studying
mutations of voltage-gated Na+ channels, but correlation
genotype-phenotype is not clear yet. We have found that
a missense mutation (M145T) on SCN1A (the alpha subunit of the voltage-gated Na+ channel) cosegregates in a
large italian family affected by simple FS. Overall, the
M145T substitution appears to determine a “loss-of-function” phenotype, suggesting a putative expression of
mutated channels in inhibitory neurons capable to
produce a network hyperexcitability that selectively causes
neuronal networks (~3 mm2, ~3000 neurons, ) by recordings from 60 electrodes, in parallel and in real time.
Excitable activity is produced by the balanced interaction
of excitatory and inhibitory neurons connected by synapses (~106), therefore it has intrinsic properties characterized by well defined statistical properties: mean
discharge frequency, correlation between neighbouring
neurons, stimulation-dependent local field potentials,
FS (Mantegazza et al., (2005) Proc. Natl. Acad. Sci.
etc. We are starting to investigate the following
102, 18177-82; Colosimo et al., (2007) Epilepsia 48:16911696). We also studied Nav1.1 Na+ channel alpha subunit M1841T mutation, found in an epileptic family charac-
problems: 1) the properties of the cortical spreading
terized by a particularly large phenotypic spectrum. The
present in KO mice for the K+ channel Kv1.1, 3) the
mutant resulted to be a loss of function because is resul-
reduced cortical activity in a mouse model of the
ted to be “trafficking-defective” (Rusconi et al., (2007)
Rett syndrome; 4) the defective neurogenesis in citro
J. Neurosci. 27:11037–11046).
kinase knockout mice.
depression in KI mice which mimics the human channelopaty of Ca2+ channels (FHM-1), 2) the epileptic seizures
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17
NICOTINIC RECEPTORS
AND VOLTAGE-GATED K+
CHANNELS IN PHYSIOLOGY
AND PATHOLOGY
PARTICIPANTS
Andrea Becchetti, Patrizia Aracri, Raffaella Morini, Silvia Consonni,
Chiara Di Resta, Paola Ambrosi, Elisabetta Cavallucci
NICOTINIC MODULATION OF THE THALAMOCORTICAL
SYSTEM
In the mammalian brain, the cholinergic fibres ascending
from the basal forebrain and mesopontine nuclei contribute to regulate the transitions between states with different
level of vigilance, including the transition between the non
rapid-eye-movement and the rapid-eye-movement phases of sleep. ACh release is also involved in the control of
synaptic plasticity and, consequently, of memory and learning. The mechanisms by which the cortical cholinergic
transmission brings about its functions are poorly understood. We are devoting particular attention to the cholinergic modulation of transmitter release and its contribution
to the regulation of the cortical functions in normal and
pathological conditions (such as sleep-related epilepsy).
We carry out patch-clamp recording in murine brain slices
and couple the electrophysiological approach with neuroanatomical and molecular biological methods.
NEURONAL NICOTINIC RECEPTORS AND SLEEPRELATED EPILEPSY
We study the properties of mutant subunits of the human
neuronal nicotinic receptors, linked to certain forms of
nocturnal epilepsy. Normal and mutant channels are
expressed in cell lines and their biophysical and pharmacological properties studied in patch-clamp. In addition,
we will address the nicotinic modulation of the thalamocortical function in murine models of these pathologies, by
applying the approaches outlined in the previous paragraph.
MOLECULAR COMPLEXES AND SIGNALING BETWEEN
INTEGRIN RECEPTORS AND ION CHANNELS
By mediating cell adhesion to the extracellular matrix,
integrins regulate many developmental processes in the
broadest sense (from cell choice between differentiation
and proliferation, to tissue remodeling and organogenesis). Ion channels would appear instead to be better suited
for rapid cellular signalatory tasks. Increasing evidence
shows however that considerable cross-talk occurs between integrins and ion channels, mediated by direct (i.e.
formation of macromolecular complexes) or indirect interaction (e.g. through G proteins). In addition, ion channel
stimulation frequently controls integrin activation or
expression. The study of channel-integrin interplay has
important mechanistic implications for understanding how
the extracellular matrix regulates as disparate processes
as muscle excitability, synaptic plasticity and lymphocyte
activation, just to mention a few. The derangement of these
processes has clear implications for pathogenetic processes, such as tumour invasivity and neurology.
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CARDIAC CELL
PHYSIOLOGY
18
PARTICIPANTS
Antonio Zaza,
Marcella Rocchetti, Claudia Altomare,
Matteo Alemanni, Riccardo Chisci,
Stefano Marangoni, Francesca Menduni.
The research of the cardiac cell physiology group is centered on the ontogenesis and modulation of myocardial
excitation-contraction (EC) coupling. The research activity
in 2007 was articulated in the following projects.
EVALUATION OF FUNCTIONAL DIFFERENTIATION IN
STEM-CELL DERIVED CARDIOMYOCYTES
In this study we tested the possibility to obtain information
on the functional differentiation of precursors by imaging
methodologies which could be applied to cell populations.
The strategy is the search of muscle-specific Ca2+ signaling, triggered by suitable agonists (caffeine, ATP etc), in
populations of precursor-derived cells. This was implemented, through the use of Ca2+-sensitive fluorescent
dyes, by counting the number of cells responding to agonist challenge in wide-field confocal images. We developed an image-analysis software to automatically count
the responsive cells and study the time course of the Ca2+
response in individual cells. The frequency of Ca2+ responding cells and the pattern of Ca2+ responses in a population was then matched with the expression of molecular
markers of muscle differentiation in the same population.
The approach developed in this investigation may be suitable to identify early functional differentiation toward
muscle phenotype and will be applied to test the differentiation of specific cell populations.
MODULATION OF MYOCARDIAL EC-coupling BY THE
PI3K/Akt PATHWAY
Recent observations indicate that the PI3K/Akt signaling
pathway is deeply involved in controlling the cardiac ECcoupling machinery. We tested the possibility to modulate
the PI3K/Akt pathway by molecules specifically designed
to bind the pleckstrin-homology (PH) domain of Akt protein, thus obstructing Akt recruitment and activation. This
is an innovative approach with still undefined functional
consequences. For this purpose we used two chemically
unrelated PH-domain antagonists, obtained through a
collaboration with the organic chemistry group of this
Department (Prof. Cipolla and Nicotra) and with a company (Nerviano Medical Science). The effects of these com-
pounds on myocyte contractility and Ca2+ handling were
compared to those of highly selective Akt1 silencing by the
RNA-interference technique. The results obtained show
that the compounds exert significant effects on contractility by cooperatively interacting with‚ ß-adrenergic receptor stimulation and that these effects are indeed mediated
by Akt modulation. This study identifies a novel target for
pharmacological modulation of cardiac function.
EFFECTS OF CHRONIC HYPOXIA ON MYOCARDIAL
ELECTROPHYSIOLOGY
Chronic hypoxia is common in respiratory diseases, a
condition in which secondary myocardial involvement is
common. Moreover, chronic hypoxia results from
uncompensated heart failure and might contribute to its
evolution. The aim of this project is to study the effects of
chronic (4 weeks) exposure of rats to hypoxia (10% O2) on
the physiology of cardiac myocytes. This might lead to
identification of molecular targets suitable to pharmacological prevention of the myocardial effects of hypoxia.
Myocytes isolated from the right ventricle (mechanically
overloaded by pulmonary hypertension), were compared
to those of the left ventricle (not mechanically overloaded). The studies initially focused on the expression of
the “late Na+ current” (INaL), which is enhanced by acute
hypoxia and may contribute to both electrical and contractile derangements. The results obtained so far indicate that the hypoxia protocol used caused marked right
ventricular hypertrophy without clear-cut derangements
on left ventricular function. Although the effects of hypoxia on electrical activity (action potential) differed between the right and left ventricles, this was not accompanied by changes in INaL. Preliminary biochemical observations, obtained through collaboration with Prof Meneveri
(Milano-Bicocca Faculty of Medicine), also suggest that
the pulmonary tissue expression of eNOS was increased
by hypoxia. This project will continue with the identification of the molecular events responsible for action
potential abnormalities and with evaluation of contractility and the Ca2+ handling function.
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19
ECOLOGY OF MARINE
AND MIGRANT BIRDS
PARTICIPANTS
Roberto Ambrosini, Pietro Bertoglio.
Maternal effects comprise a class of phenotypic effects
where the genotype of a mother is expressed in the phenotype of her offspring, unaltered by paternal genetic
influence via e.g. molecules deposited by the mother in
the egg cell. We are currently studying maternal effects
mediated by carotenoids content in the eggs of the yellow-legged gull (Larus michahellis).
Migratory connectivity is a new ecological concept that
refers to the extent of the connection between the areas
where populations spend different phases of their
annual life-cycle. We have developed a novel method for
quantifying migratory connectivity that may have broad
applications in studies of migratory systems and in conservation projects.
Phenology is the study of the times of recurring natural
phenomena. There is compelling evidence of significant
temporal changes in the phenology of bird migration,
that are probably linked to recent climate change.
Currently, we are developing a large collaborative project with the aim to establish a database of European
bird arrival/migration dates to promote research and
dissemination of scientific knowledge concerning phenological changes.
IDENTIFICATION AND ANALYSIS OF THE MOLECULAR BASIS AND PREDISPOSING FACTORS OF
IDIOPATHIC EPILEPSIES
20
PARTICIPANTS
Romina Combi
Epilepsy is a common and devastating neurological disorder in which genetic background and physiopathological
mechanisms underlying the clinical phenotype are not
fully characterized yet. Recent data indicate the existence
of several mutations in voltage dependent ion channels,
neurotransmitters receptors and other types of proteins
involved in determining epilepsies. However, these gene
discoveries have been in epilepsies with Mendelian
modes of inheritance, which comprise only a tiny fraction
of all epilepsy. Even in these forms (of both adult and paediatric age), a high degree of complexity is often displayed.
As in other complex inheritance diseases, this variability
is likely to result from the modifying effects of other genes
or environmental factors. By means of an integrated clinical and molecular approach (comprising genetic counselling, DNA analysis, DNA sequencing, linkage analysis), we
study a large cohort of families affected by different forms
of epilepsy searching for new genes and new mutations
involved in the pathogenesis of such diseases. Moreover,
we perform functional in vitro studies to evaluate the
effect of the identified mutations.
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ZOOLOGY
AND ANIMAL BIOLOGY
21
PARTICIPANTS
Aldo Zullini, Maurizio Casiraghi,
Michela Barbuto, Andrea Galimberti.
STUDIES ON INTRASPECIFIC VARIABILITY IN FREELIVING NEMATODES
The freshwater nematode Eumonhystera dispar is studied in specimens from both standing and running
waters. They present slightly different molecular features (at ITS1 ribosomial region) and also parallel somatometric differences (detected by Principal Component
Analysis studies). The aim of this project is to assess biogeographic insulation among different populations and
possibly micro-evolutive trends and adaptations.
Moreover, two species of Eumonhystera were studied by
18S rDNA to assess their phylogenetic relationship with
other nematodes.
Moreover we examined nematodes from mosses of different ecological conditions from biocenotic and genetic
point of view.
STUDIES ON ALPINE SPRINGS INHABITING NEMATODES
A taxonomic and ecological study of freshwater nematodes collected in 96 Alpine springs by the Trento Natural
History Museum is in progress. The collected data will be
correlated with other biological (algae, protozoa, molluscs, aquatic insects, etc.) and non biological parameters
to assess the ecological status and quality of these mountain springs. Such microhabitats are an important feature
of the Alpine environment.
DNA BARCODING: A LINK BETWEEN BASIC AND
APPLICATIVE SCIENCES
Ever since 250 years taxonomy has played a key role in
biological studies. However, only a little portion of the
living beings present on the earth has been identified and
formally described, while the study of taxonomy in the
universities and the financial supports to taxonomic studies is decreasing year after year. To renew taxonomy, the
main challenges are informatization and molecularization. The informatization has the aim to allow a better
and broader access to the taxonomic results. Molecular
taxonomy has the aim to provide, even to a not taxonomist,
a rapid and sure way to discriminate organisms on the
basis of the variability of some molecular markers
(mitochondrial genes in almost all the metazoans), a
method known as DNA Barcoding of living beings.
Given these considerations, the goal of our research
project is to develop a DNA Barcoding approach for
different organisms. The molecular approach will be
then compared to the traditional taxonomy, to allow, or
not, its validation.
THESE FOLLOWING ARE OUR RUNNING PROJECTS ON
DNA BARCODING:
1) Food tracking: on fish (in particular part of fish) in
collaboration with the Milan Fish Market and the Nucleus
Antisophistication of the Corps of Carabinieri.
2) Nematode symbioses: using filarial nematodes and
their endosymbionts (Wolbachia) as target for our analyses.
3) Free-living nematodes: analysing natural population of
free-living nematodes hosted in different habitats
(i.e. water, moss, soil).
4) Birds: studying populations of non-autochthon species
of birds.
5) Bats: studying national populations of bats species.
6) Fish and their parasites: to reconstruct the possible
coevolution of host and symbionts, and their population
structure and speciation.
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22
FRESHWATER
AND MARINE ECOLOGY
PARTICIPANTS
Paolo Galli, Fabrizio Stefani,
Francesca Benzoni, Giovanni Strona.
A HOST-PARASITE MODEL FOR THE DISPERSAL OF
LESSEPSIAN SPECIES IN MEDITERRANEAN.
The 1869 opening of the Suez Canal created a direct link
between Mediterranean and Red Sea, allowing the entry
into the Levantine aquatic system of non-native species,
particularly from Erythrean basin, process that has
accelerated in the recent years concurrently to the warming trend of the seawater. Among fish Siganus luridus,
has proven to be extremely successful in colonizing a
large part of Eastern Mediterranean coasts until Linosa
Island, that constitutes the western boundary of the species distribution. The aim of the work is to provide a
theoretical framework, through a metapopulation model,
to explore alternative assumptions on the Lessepsian
invasion by using information on the presence of fish
parasite as fingerprint of the adult host arrival time. In
the model, host populations are divided into identical
interconnected sub-populations that are linked by
dispersal and well-mixed with respect to parasite
transmission.
MONOGENOIDEA OF RED SEA.
Monogenea with more than 24.000 species (Whittington,
1998) represents one of the main group of fish parasites.
Although Monogenoidea species are abundant, few studies have been carried out in the Red Sea. The primary
purpose of the project is to provide an update of the knowledge on monogenoids dispersion in the area of Ras
Mohammed National Parks. In particularly the goals
are: 1) realisation of check-list of monogenoids from
native fish; 2) realisation of check-list of monogenoids
from endemic fish; 3) creation of a parasite collection.
ENVIRONMENTAL AND BIOLOGICAL FACTORS AFFECTING JUVENILE AND ADULT CORAL COMMUNITIES IN
KUWAIT OFF-SHORE REEFS.
The Arabian Gulf is a shallow sedimentary basin where
environmental conditions can be limiting for coral reef
growth. Such conditions include, among others, extre-
mes (both high and low) in seawater temperature regime and the Shatt al Arab freshwater input. a waterway
into which the Tigris, Euphrates and Karun rivers
discharge their waters. Coral reefs fringing the offshore islands of Kubbar, Qaro and Um Al Maradeem off
the southern shores of Kuwait are the most northerly
reefs in the Arabian Gulf. These shallow high-latitude
reefs are characterized by a low coral species diversity,
and a relatively high cover dominated by Acropora spp.
and Porites spp. paucispecific assemblages, a typical
condition in other high-latitude reefs in the Arabian
region. A survey of kuwaiti off-shore coral reefs was carried out in order to assess coral community structure,
and adult and juvenile coral diversity and distribution. A
survey of the abundance and distribution of the sea
urchin Echinometra mathaei was also conducted in
order to investigate possible relationships between the
density of coral juveniles, and grazing sea urchins.
EFFECTS OF BEDOUIN ARTISANAL FISHERIES ON
CORAL COMMUNITIES STRUCTURE AND COMPOSITION
IN THE NABQ MANAGED RESOURCE PROTECTED AREA,
SOUTH SINAI, EGYPT.
The Nabq Managed Resource Protected Area (MRPA) is
located on the southern part of the Gulf of Aqaba coast
of South Sinai, Egypt. A network of four no-take zones
(NTZs) was established in 1995 within the Nabq MRPA to
promote sustainable management of finfish stocks
exploited by artisanal Bedouin fishermen. Despite the
MRPA and NTZs establishment, no published account of
coral reef zonation and benthos composition exist for
Nabq. Gathering of Tridacna spp. and of other edible
invertebrates on the reef-top is also part of the traditional fishery practised by the local Bedouin population,
mainly by women. To assess the effects of these NTZs on
gleaned invertebrates a survey of Tridacna and other
invertebrates, along with reef structure and coral community composition, was undertaken within NTZs boundaries, and in four adjacent fished areas.
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PLANT MOLECULAR AND
CELLULAR BIOLOGY
23
PARTICIPANTS
Paolo Crosti, Massimo Labra,
Massimo Malerba, Fabrizio De Mattia, Ilaria Bruni,
Fabrizio Grassi e Nicla Contran.
PLANT BIODIVERSITY AND MOLECULAR MARKERS
PROGRAMMED CELL DEATH (PCD) IN PLANTS
Ilaria Bruni, Fabrizio De Mattia, Fabrizio Grassi, Massimo Labra.
Paolo Crosti, Massimo Malerba, Nicla Contran.
DNA molecular markers represent a suitable tools to
investigate plant genetic diversity. Our group uses these
tools to analyze Vitis vinifera genome to define the
relationships among different cultivars and to elucidate grapevine domestication processes. Considering that
the most relevant differentiating trait between wild and
cultivated grapevine is the mating system our investigation
was also directed on genes involved in sexual organs and
flower development.
Programmed cell death (PCD), a genetically controlled
process, plays a pivotal role in several developmental
processes of plants and is involved in responses to
environmental stresses and in defence mechanisms
against pathogens. Researches to elucidate the basic
mechanisms of plant PCD are in rapid expansion.
During the last years we studied this process in Acer
pseudoplatanus L. cultured cells by means of the phytotoxin fusicoccin (FC). A role for mitochondrion as integrator of cell stress and regulator of PCD has been proposed for plants too; in fact cytochrome c (cyt c) release
is a common event in several PCD of plants, including
FC-induced PCD. The mechanism of cyt c release in
plant mitochondria is poorly understood. In the last year
we investigated this process by means of the immunosuppressive drug cyclosporin A (CsA), which allows to
discriminate between two different mechanisms of cyt c
release. Our results suggest that the FC-induced cyt c
release occurs through a CsA-sensitive system, similar
to the permeability transition pore of animal mitochondria.
In the case of spontaneous plant our work was focused
on the Rhododendron species dispersed in the Alps and
Apennines. In order to clarify the phylogenetic relationships among the close related species of Rododendron
and to define the genetic variability among different
populations our research programme was focused on
the set up of different DNA markers suitable for different
Rhododendron genome.
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COMPUTATIONAL INVESTIGATION
OF STRUCTURE-ACTIVITY
RELATIONSHIPS IN PROTEINS
AND BIOMIMETIC
COMPLEXES
24
PARTICIPANTS
Piercarlo Fantucci, Luca Bertini,
Denis Bilotta, Luca De Gioia,
Francesca Falcetta, Elena Papaleo,
Marco Pasi, Giuseppe Zampella.
DFT INVESTIGATIONS OF METALLO PROTEINS AND
BIOMIMETIC METAL COMPLEXES
Luca Bertini, Luca De Gioia, Piercarlo Fantucci, Giuseppe Zampella.
The project is aimed at elucidating both the activity
mechanism and the stereo-electronic properties of some
active sites in metalloenzymes, as well of the key regions
of proteins, involved in their biological role. Effort is put in
determining the chemical features which characterize a
transition metal when bound to the polypeptide. Ab initio
Density Functional Theory (DFT) approaches are used in
order to compute the electronic structures and perform a
detailed analysis of models employed to simulate the biosystems under study.
COMPUTATIONAL INVESTIGATIONS OF STRUCTUREACTIVITY RELATIONSHIPS IN PROTEINS
Luca De Gioia, Piercarlo Fantucci, Elena Papaleo, Marco Pasi, Giuseppe
Zampella.
Molecular dynamics simulations and homology modelling are used as main techniques with the aim of investi-
gating structure-function relationship in enzymes and
proteins. In fact, long and multiple simulations of biomolecular systems can allow obtaining insights into biomolecular processes at the atomic level, which are often
hardly accessible to experimental methods. Attention is
addressed to the effect of the temperature on protein
stability and the interaction between enzymes and their
cofactor or some inhibitors.
DEVELOPMENT OF BIOINFORMATICS TOOLS FOR
ANALYSIS OF PROTEINS AND THEIR POST-TRANSLATION
MODIFICATIONS
Luca Bertini, Denis Bilotta, Francesca Falcetta, Piercarlo Fantucci.
In order to overcome the limitations of proteomic techniques to determine post-translational modifications
(PTM), computer programs have been developed to analyze amino acid sequences for PTMs and compute modifications of molecular mass and isoeletric point. The
structural bases of S. pyogenes pyrogenic exotoxin B
activation are under study, and the design of specific
inhibitors is underway.
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DESIGN, SYNTHESIS
AND MOLECULAR RECOGNITION STUDIES
ON BIOACTIVE COMPOUNDS
25
PARTICIPANTS
Francesco Nicotra, Laura Cipolla, Barbara La Ferla,
Cristina Airoldi, Maria Gregori, Paolo Galliani,
Davide Bini, Valerio Spinosa.
The area of investigation of the research group ranges in
the field of design, synthesis and biological evaluation of
bioactive compounds and their conjugates. Particular
attention is devoted to the generation of inhibitors, agonists and antagonists not only as new lead compounds in
drug research, but also as tools to understand unknown
biological pathways (chemical genomics studies).
SYNTHETIC TARGETS FOCUSED IN 2007 ARE:
• Inhibitors of bacterial LPS biosynthesis as potential
antibacterial agents
• Inhibitors of Protein Kinase B as potential antitumor
agents and cardiac modulators
• Inhibitors of glycosidases as potential antiviral
agents and metabolic diseases regulators
• Cholera Toxin antagonists based on D-galactose
scaffolds
• Drugs fused into glycidic structures, in particular
glyco-benzodiazepines and GABA-receptor antagonists,
in order to modulate the pharmacokinetic and the
conformational properties
The conjugation of drugs to specific ligands or transporters for drug delivery and drug targeting studies has also
been performed, and the possibility to generate clusters
of drugs/ligands in order to increase the affinity exploit
the multivalency phenomenon has been studied.
NMR STUDIES ARE PERFORMED FOR:
• Structure elucidation
• Conformational analysis
• Epitope mapping studies (ligand-receptor
interactions studies at atomic level)
• Adhesion kinetic studies
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26
BIOORGANIC AND
MEDICINAL CHEMISTRY
PARTICIPANTS
Francesco Peri, Alessandro Palmioli, Matteo Piazza, Clara Rossini,
Anna Paola Chiarlone, Silvia della Fiorentina, Cristina Airoldi.
Synthesis of bioactive and pharmacologically active
molecules and investigation of protein-ligand interaction.
SUGAR-DERIVED RAS PATHWAY INHIBITORS
We are interested in the synthesis of novel molecules that
are able to interfere with the signal transduction pathway
associated to human Ras proteins. We developed some
small molecules that are able to bind human p21hRas
protein. As constitutively active Ras mutants are responsible of the generation and growth of about the 30% of
human tumor (in particular prostatic and colorectal cancers), small organic molecules that bind and deactivate
Ras are potential highly selective antitumor drugs. The
new compounds developed in our lab have been synthesized starting from natural sugars such as glucose and arabinose and tested in vitro their capacity to bind Ras and to
inhibit GTP/GDP exchange. We found a nucleotide exchange inhibition in vitro. We are also testing our compounds
on cell lines that are representative of Ras-dependent
tumors (such as the HCT-116 colorectal cancer cell line).
We have very recently collected promising results on the
selective toxicity of our compounds on cell lines containing the oncogenic Ras mutant G13D. Our aim is to elucidate the molecular mechanism of Ras inhibition. This project is highly interdisciplinary and is developed in collaboration with the following research units: Biochemistry and
Biology of Ras, group of prof. Enzo Martegani and Marco
Vanoni; Cell biology, Oncology, group of prof. Alberto
Bardelli (ISMCC, Candiolo and University of Torino); molecular modelling: group of prof. Luca de Gioia (Unimib).
NOVEL MONOSACCHARIDES ACTIVE AS LIPID A ANTAGONISTS ON THE TLR-4 RECEPTOR
We are developing a new class of compounds derived
from the monosaccharide D-glucose that are able to inhibit the signal pathway associated to the TLR-4 receptor.
Bacterial lipopolysaccharides (LPS) and their bioactive
portion, the lipid A, bind to TLR-4 initiating the signal
cascade that causes cytokine production. Several inflammatory, autoimmune and tumoral syndromes as well as
sepsis and septic shock depend on the activation of TLR-
4 pathway. Our compounds have been patented as hits for
the development of innovative anti-inflammatory and
anti-sepsis drugs. We are refining the structure of our
molecules, taking advantage from the very recently published crystal structure of TLR-4 with an inhibitor (eritoran)
in the hope to obtain a new generation of highly selective
TLR-4 agonists or antagonists. We are using our compounds to shed light on the still unclear mechanism of
TLR-4 receptor complex activation and consequent signal
transduction. This project is developed in collaboration
with: prof. Francesca Granucci (Immunology), Dr. Barbara
Costa (Pharmacology), Dr. Paola Fusi (Biochemistry),
prof. Theresa Gioannini (University of Iowa, USA)
(Biochemistry, binding experiments with purified receptors).
NEW DENDRIMERIC MOLECULES FOR MULTIPLE ANTIGEN PRESENTATION AND SIGNAL AMPLIFICATION
We are developing in collaboration with Diasorin S.p.A.
(Nerviano, MI) new molecules that present several copies
of clinically important antigens and of luminescent molecules (isoluminol derivatives). These compounds, with a
tree-like shape (dendrimers) will be used as components
of kits for the detection of antigens in the fluids of
patients. The multiple presentation to immobilized antibodies of the antigen and the presence of several signalgenerating units in the molecules, should ensure signal
amplification that is very important for the sensitivity and
reliability of immunochemistry tests. This project is in collaboration with the immunology and immunochemistry
unit of Diasorin S.p.A.
CHEMOSELECTIVE GLYCOSYLATION METHODS FOR THE
GENERATION OF BIOACTIVE MOLECULES
We are investigating new chemical strategies to glycosylate bioactive compounds in a rapid, chemoselective and
efficient way. We are designing new reversible glycosidic
bonds to link sugars to pharmacologically active compounds and have access to pro-drugs with improved bioavailability and pharmacokinetic.
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MOLECULAR MODELLING
AND COMPUTATIONAL
CHEMISTRY
27
PARTICIPANTS
Giorgio Moro, Gloria Saracino,
Flavio Amara.
Computational approaches based on Molecular Dynamics
simulations, Quantum Mechanical methods and 3D
Quantitative Structure-Activity Relationships are employed
to study biological processes at the molecular level. The
computational approach taken in our research on biological processes focuses mainly on three methodological
areas. One includes a variety of methods based on
Molecular Mechanics (MM) and Molecular Dynamics (MD).
The second is an approach based on advanced Quantum
Mechanical (QM) methods applied to model systems. The
third is an approach aimed at obtaining statistical models
through an analysis of data inferring relative Quantitative
Structure-Activity Relationships (QSAR).
As is well known, approaches based on MD theories are
the only ones presently available to study complex systems
like proteins in solution. The approach to the problem of
protein structure at the classical level is even more acute
when there is the modelling of interaction between proteins themselves, between protein and DNA fragments or
between protein and substrates (as in drug discovery, toxicology studies or virtual enzyme engineering).
However, MD methods are not completely free of difficulties, which are generated just by the very high number of
degrees of freedom (about 105). In practice it is impossible
to sample the phase space exhaustively due to the limitations in reliability of the final results. Given our awareness
of the difficulties involved, we took great care when
applying the MD to maximize the degree of phase space
sampling, using the repeated trajectory technique, the
essential dynamics technique extensively, in order to
extract the low frequency motions of biological relevance,
and the repilca exchange technique to overcome the
potential energy holes problem. These studies are carried
out in collaboration with U. Cosentino (Dept. of Science of
Environment and Territory, UNIMIB)
SPECIFIC TOPICS OF INTEREST ARE:
• Properties of prion protein peptides (collaborations
with A.Villa - Max-Planck-Institute for Polymer
Research – Mainz – Germany; M. Salmona – Istituto
Mario Negri – Milano)
• Thermal stability of the Sulfolobus solfataricus
Caroxypeptidase active site (collaborations with P.
Tortora - Dipartimento Biotecnologie e Bioscienze)
• Characterization of a new contrast agent for selective
targeting in Magentic Resonance Molecular Imaging
(collaborations with F. Nicotra and L. Cipolla –
Dipartimento Biotecnologie e Bioscienze)
• Interaction of the HIV-1 viral protein R with the adenine
nucleotide translocator protein
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28
OUTER MEMBRANE
BIOGENESIS IN
ESCHERICHIA COLI
1
PARTICIPANTS
Alessandra Polissi. Paola Sperandeo,
Silvia Sommaruga, Riccardo Villa.
2
1_Escherichia coli
Rod-shaped Bacterium
with Multiple Flagella
2_Escherichia coli strains
undergoing conjugation
The cell envelope of Gram-negative bacteria represents
an effective permeability barrier against external noxious
agents and cell envelope components are primarily involved in host colonization or infection. However many
aspects of cell envelope biogenesis remain still obscure.
A peculiar structure of Gram-negative envelope is the
outer membrane an asymmetric lipid bilayer with phospholipids and LPS forming the inner and outer leaflet,
respectively. LPS is a complex essential molecule relevant
to initial bacterial attachment, evasion of host defenses,
and establishment of infection. Despite structure and
composition of the OM have long since been known, many
aspects of its biogenesis still remain obscure.
MOLECULAR MECHANISMS OF LPS TRANSPORT TO THE OM
Alessandra Polissi, Paola Sperandeo, Riccardo Villa.
Genetic and biochemical approaches are being used to
identify new proteins implicated in the LPS biogenetic
pathway and to study how these proteins interact. By dissecting the mechanisms of LPS transport and identifying
new components involved we aim at obtaining a deeper
knowledge of outer membrane biogenesis, a fundamental
process for bacterial cell life and pathogenicity. This not
only will allow a better understanding of the mechanisms
that control bacteria-host interactions but is also a prerequisite and a significant step forward to the second objective of this research. Collaboration with Gianni Dehò University of Milano and Thomas Silhavy - University of
Princeton.
THE LPS BIOGENETIC PATHWAY AS TARGET FOR THE
DESIGN AND SYNTHESIS OF NOVELS ANTIBACTERIALS
Alessandra Polissi, Paola Sperandeo, Silvia Sommaruga.
Structural and functional studies of target proteins known
to play key roles in the biogenesis of LPS are currently
ongoing. The information will be used to design and synthesize novel lead compounds that inhibit the LPS biogenetic pathways in the hope to develop new therapeutic
strategies against infectious diseases. Collaboration with
the group of Francesco Nicotra, UNIMIB and Martino
Bolognesi, University of Milano.
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INDUSTRIAL BIOTECHNOLOGY:
ADAPTATION OF THE MICROBIAL CELL FACTORY
TO TECHNICAL CONSTRAINTS
29
PARTICIPANTS
Danilo Porro, Luca Brambilla, Paola Branduardi, Gianni Frascotti,
Simone Passolunghi, Tiziana Fossati, Laura Dato, Dario Losio,
Valerio Mezzasalma, Roberto Pagani, Luca Riboldi, Giusy Adamo, Valerio Balivo,
Alessio Casati, Monica Corbetta, Lorenzo Ferrera, Marco Lattanzi, Riccardo
Posteri, Giorgia Rossi, Irene Sberna.
Evolution has produced a huge variety of organisms living
in radically different environments. Some of these organisms have evolved metabolic pathways leading to the
synthesis of potentially useful compounds that are difficult to produce by the chemical industry or that are environmentally harmful to manufacture. It has to be reminded that the fundamental basis of evolution is the need to
survive and reproduce, not to produce potentially important and commercially valuable products. Indeed, interesting proteins and metabolites are very often produced by
wild type organisms in such low concentrations that biotechnological exploitation is today still impractical.
rDNA platforms allow, sometimes in a quite simple way,
the development of new micro-organisms leading to the
production of new products. The existing rDNA applications for eukaryotic microbial hosts are the results of less
than three decades of global experience developing processes for the production of heterologous proteins, fine
chemicals, vitamins, nutraceuticals, biofuels and animal
nutritional aids such as amino acids. Unfortunately, the
majority of the rDNA engineering processes, besides the
challenges encountered during the research and development phase, fail during the scale-up phase. Indeed, in an
industrial process, the microorganism used as a mean of
production, is exposed to several stresses that can lead to
lower production, lower productivity and lower yield of the
product. A stress is typically caused by stressors (or stimuli), i.e. agents of a physical, chemical or biological
nature that represent a change in the usual intracellular
or extracellular conditions. It is therefore highly desirable
to consider strategies for minimizing stress. In this
respect, our laboratory has developed (i) a series of cell
factories producing heterologous compounds, like proteins, enzymes, organic acids, biofuels and nutraceuticals
(ii) a series of yeast strains with improved resistance to
specific constraints imposed by the process itself and (iii)
a study and a model of the correlation between the size of
the single yeast cell and its cellular metabolism.
(I) MICROBIAL CELL FACTORIES AND MAIN PRODUCTS
For twenty five years our group has been involved in the
production of homologous and heterologous proteins in a
variety of yeast hosts, from the conventional S. cerevisiae,
to the non conventional Kluyveromyces lactis,
Torulaspora delbrueckii, Zygosaccharomyces bailii
applying different fermentative technologies (batch, continuous and fed-batch). As an example, we developed yeast
strains capable of producing organic acids from glucose
(i.e. lactic and ascorbic acid). More recently, our attention
is also focused on the production of biofuels.
(II) IMPROVING RESISTANCE IN MICROBIAL CELL FACTORIES
In order to develop an effective process of production, cell
factories not only have to produce the molecule of interest, but they also have to face the constraints often imposed by the process itself. We proved that yeast cells engineered to produce ascorbic acid acquire an increased
robustness in respect to different limiting conditions such
as low pH, oxidative stress and the presence of high concentrations of organic acids. In addition, said resistance
can be achieved also by modulating other key elements.
(III) PHYSIOLOGICAL AND MODELLING STUDIES OF THE
CELL FACTORIES
The control of both metabolism and cell cycle progression
by the cellular environment has a key role in the regulation of growth and cell proliferation and production in all
organisms. Specific attention has been devoted to study
and to model the correlation between the size of the single yeast cells and its metabolism.
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3
SCIENTIFIC
PUBLICATION
INDEX , GRANTS
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RESEARCH GRANTS AND CONTRACTS
3.1
ALBERGHINA L. Rete Italiana di Bioinformatica
(ItalBioNet). – FIRB-MIUR
ALBERGHINA L. Eukaryotic unicellular organism biology
– systems biology of the control of cell growth and proliferation (UNICELLSYS). – European Commission
BARABINO S. A role for the pre-mRNA processing factor
CF Im in quality control of mRNA function. – PRIN 2006,
MIUR
BARABINO S. Genomic and Proteomic Analysis of PremRNA Processing in Amyotrophic Lateral Sclerosis. –
Fondazione Cariplo
BECCHETTI A. Recettori nicotinici cerebrali e patologie
epilettiche. – PRIN 2005 MIUR
BECCHETTI A. Functional Test of Ion Channels. –
Telethon (GTF03007)
BECCHETTI A. Recettori nicotinici cerebrali e patologie
epilettiche. – BML Foundation
CASTAGNOLI P. Integrated functional genomics in
mutant mouse models as tools to investigate the complexity of human immunological disease. – European
Commission
CASTAGNOLI P. Molecular Basis of vascular events leading to thrombotic stroke. – European Commission
CASTAGNOLI P. Molecular markers of M. tuberculosis
interactions with host phagocytes. – European
Commission
CASTAGNOLI P. Microbial Action on immune survival. –
European Commission
CASTAGNOLI P. Microarrays a DNA per lo studio della
variabilita genetica: Piattaforme micro e nanotecnologiche per diagnostica medica avanzata e nuove procedureterapeutiche. – FIRB MIUR
CASTAGNOLI P. Identificazione dei pathways molecolari
indotti nelle cellule dendritiche da stimoli promotori della
polarizazzione dei T linfociti verso un fenotipo funzionale
Th1 o Th2. – COFIN MIUR
CASTAGNOLI P. Bioinformatic tools for identifying,
understanding and attacking targets in cancer. – AIRC
Bioinformatics Center Grant/ BICG
DE GIOIA L. Basi molecolari dell'adattamento alle basse
temperature degli enzimi da organismi antartici. –
Progetto Nazionale di Ricerca in Antartide, CNR
DOGLIA S.M. Processi di funzionalizzazione di polimeri
per la modifica della biocompatibilità e della adesione di
proteine. – Fondazione Cariplo
FOTI M. Generation of a coronavirus-based multigene
AIDS vaccine and evaluation in a preclinical siv model. –
European Commission
FUSI P. Sialidasi umane: biologia strutturale, biochimica
funzionale e implicazioni patologiche. – PRIN 2006 MIUR
GALLI P. Strategie da adottare per migliorare e tutelare le
stato ecologico della fauna ittica. – Agenzia Regionale per
la Protezione dell’Ambiente della Lombardia
GALLI P. Modellizzazione della produzione primaria fitoplanctonica nel mediterraneo in rapporto alla distribuzione dei cetacei. – Consorzio Interuniversitario per lo
Studio del Mare
GIAGNONI G. Effect of transplantation of the human stem
mesenchymal cells and of mGlu2/3 agonists on the
expression and level of pro- and anti-inflammatory cytokines in dorsal root ganglia and in spinal cord of neuropathic mice. – PRIN MIUR
GRANUCCI F. Meccanismi di induzione di tolleranza in
cellule T auto reattive coinvolte nella risposta autoimmune presente nella Cheratite Erpetica Stromale. –
Fondazione Cariplo
GRANUCCI F. Key regulators of DC-primed anti-tumor
NK cell functions. – AIRC
GRANUCCI F. Dendritic cells for novel immunotherapies.
– European Commission
LABRA M. Salvaguardia della biodiversita’: conservazione
ex situ delle piante lombarde a rischio estinzione. Fondazione Banca del Monte di Lombardia
LONGHESE M.P. Genetic integrity maintenance: interrelationships between DNA damage checkpoints and
telomere metabolism. – AIRC/2005
LONGHESE M.P. Genetic integrity maintenance: interrelationships between DNA damage checkpoints and
telomere metabolism. – AIRC/2006
LONGHESE M.P. Identificazione e caratterizzazione di
geni coinvolti nel mantenimento dell'integrità del genoma e nella prevenzione della cancerogenesi. –
Fondazione Cariplo 2005-2007
LONGHESE M.P. Meccanismi di controllo dell'integrità
dei cromosomi. – PRIN 2005-2007, MIUR
LOTTI M. Valorizzazione delle risorse biologiche.
Sviluppo di nuove tecnologie per l’ dentificazione, caratterizzazione e produzione di molecole di interesse farmaceutico e industriale presenti nelle Brassicacee. –
Projects for Industrial Research MIUR.
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LUCCHINI G. Mantenimento dell'integrità genetica: interrelazioni fra checkpoints da danni al DNA e metabolismo
telomerico. – PRIN 2005-2007, MIUR
variegated aneuploidy (MVA) genetic disease by studying
the mechanisms controlling aneuploidy occurrence in the
budding yeast model system. – Telethon
MARTEGANI E. Coinvolgimento delle GTPasi Ras e Ral
nella proteolisi della proteina precursore del peptide
beta-amiloide e nella patogenesi del morbo di Alzheimer
– PRIN 2005 MIUR
PIATTI S. Identificazione e caratterizzazione di geni coinvolti nel mantenimento dell’integrità del genoma e nella
prevenzione della cancerogenesi. – Fondazione Cariplo
MARTEGANI E. Purificazione e valutazione dell’ attività
biologia di NGF ricombinante umano. – PRIMM/Blueprint
MORO G. Nuove strategie computazionali per il modelling
di nano-agglomerati vetrosi e delle loro proprietà spettroscopiche. – PRIN 2006, MIUR
NICOLIS S. Ruolo e meccanismi d'azione del fattore trascrizionale Sox2 nelle cellule staminali neurali: un
approccio genetico mediante ablazione condizionale di
Sox2 nel topo. – MIUR 2005
NICOLIS S. Il fattore trascrizionale Sox2 nella genesi e
mantenimento delle cellule staminali neurali e dei neuroni: ablazione condizionale di Sox2 in topo – Fondazione
Cariplo
NICOLIS S. NS-toolkit - A genetic toolkit for the analysis
of mouse neural stem cells. – Cariplo NOBEL.
NICOLIS S. An in vivo genetic approach to the role and
mechanisms of action of the transcription factor Sox2 in
neural stem cells: conditional cell type-specific, developmentally inducible deletion of Sox2. – Telethon 2005
NICOTRA F. Sistemi di separazione ad elevate prestazioni basati sul riconoscimento molecolare chemo- e stereoselettivo. – PRIN 2005, MIUR
NICOTRA F. Piattaforma integrata per la progettazione e
la produzione high throughput di enzimi e peptidi ingegnerizzati. Valutazione della loro attività biologica rispetto a specifici substrati molecolari di interesse farmaceutico, con riferimento allo screening di prodotti oncologici
ed alla produzione di antibiotici e nutracetici (PANDA). –
Metadistretti Regione Lombardia
POLISSI A. Antibacterial properties of Silver nanoparticles. – Menphis S.p.A.
OTTOLENGHI S. “Programmi genetici” comuni a vari tipi
di cellule staminali: uno studio in cellule staminali cardiache e in cellule germinali. – MIUR 2005
OTTOLENGHI S. Genomica e proteomica funzionale per la
prevenzione della patologia ematologica nelle emoglobinopatie ereditarie, in particolare le talasemie. –
Fondazione Cariplo
RONCHI A. Genomica funzionale della transizione
embrionico-adulta nell'ematopoiesi. – MIUR
TORTORA P. Transcriptomics and Proteomics
Approaches to Diseases of High Sociomedical Impact: a
Technology Integrated Network. – Fondazione Cariplo
(N.O.B.E.L. project)
TORTORA P. Network tecnologico integrato per lo studio
proteomico e trascrittomic odi malattie neurodegenerative correlate a deposizione di amiloidi. - Ministero della
Salute/Regione Lombardia
VANONI M. Sviluppo di peptidi e loro derivati con attività
inibitoria della via di traduzione del segnale mediata da
Ras. – Creabilis
VANONI M. Sensing extracellulare ed intracellulare di
nutrienti e progressione del ciclo cellulare nel lievito
Saccharomyces cerevisiae. – PRIN MIUR
VESCOVI A. Cellule staminali neurali umane ed ingegneria dei tessuti biologici per la rigenerazione di lesioni al
sistema nervoso centrale e periferico. – Fondazione
Cariplo
NICOTRA F. Materiali innovativi per lo sviluppo di bio-protesi articolari. – FIRB MIUR
VESCOVI A. Plurigenes: De-differentiation of terminally
differentiated cells into pluripotent cells. – European
Commission
NICOTRA F. Nanoparticles for therapy and diagnosis of
Alzheimer disease. – European Commission
VESCOVI A. Neuro: Towards the neuronal machine European Commission
PERI F. Functionalized Viral Nanoparticles for multiple
antigen presentation and chemoluminescent signal
amplification. – DIASORIN S.p.A (Nerviano, MI)
VESCOVI A. Evi-Genoret: Functional genomics of the retina in health and disease. - European Commission
PIATTI S. Functions and interactions of mitotic checkpoint proteins during the cell cycle. – AIRC
PIATTI S. Unravelling the molecular bases of the mosaic
VESCOVI A. Stabilità funzionale e fenotipica delle cellule
staminali cerebrali umane: ruolo nella terapia cellulare
delle malattie neurodegenerative e neurooncologiche. ISS: Protocollo d'intesa IRCSS Istituto Nazionale dei
Tumori Conv. 69
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R ESEARCH
GRANTS AND CONTRACTS
VESCOVI A. Tumor neural stem cells in the in vitro and in
vivo modelling and studying of the adult human glioblastomas. – PRIN 2006 MIUR
ZAZA A. Analysis of Istaroxime effect on the stability of
intracellular Ca2+ stores and on Ryanodine receptors properties. – Debiopharm, Lausanne (CH)
VESCOVI A. Trapianto di cellule staminali adulte –
Regione Lombardia
ZAZA A. Analisi dell’effetto del PST2744 sulla stabilità del
deposito intracellulare di Ca2+ e sulle proprietà dei canali RyR. – Istituto di Ricerche Prassis-Sigma Tau (MI)
VESCOVI A. Tumor neural stem cells in the vitro and in
vivo modeling and studying of the adult human glioblastomas. - AIRC
WANKE E. Canalopatie dei canali voltaggio-dipendenti
Na+ e K+ nel sistema nervoso centrale: studi avanzati con
metodiche MEA (multielettrode arrays). – PRIN 2005
MIUR
WANKE E. Studio delle proprietà biofisiche di specifici
sottotipi del canale del sodi in modelli in vitro. – NewronMilano-ricerche
WANKE E. Studio di ricerche funzionali su proteine di
membrana. – Axxam-Milano Ricerche
ZAZA A. Application and process optimization of human
stem cells for myocardium repair" (SC & CR). – European
Commission
ZAZA A. Investigation of cardiotoxicity related to commonly inhibited signaling pathways in cancer cells. –
Nerviano Medical Science (MI)
ZAZA A. Role of the “late Na+ current” in myocardial and
neuronal effects of chronic hypoxia. - CVT Therapeutics,
Palo Alto CA (USA)
ZAZA A. Funzione del reticolo sarcoplasmico e stabilità
del deposito di Ca2+ nel muscolo cardiaco. – Interlink
Montpellier (II04C570GL), France
ZULLINI A. Applicazione del barcoding molecolare ai
nematodi zooparassiti (Spirurina) e integrazione con la
tassonomia tradizionale. – Progetto Galileo – Università
Italo-Francese
S EVERAL YOUNG SCIENTISTS WERE SUPPORTED THROUGH THE PROJECT P ROGETTO I NGENIO GRANTED BY F ONDO
S OCIALE E UROPEO , M INISTERO DEL L AVORO E DELLA P REVIDENZA S OCIALE AND R EGIONE L OMBARDIA
PUBLICATIONS
3.2
AINA R, LABRA M, VANNINI C, MARSONI M, CUCCHI U,
BRACALE M, SGORBATI S, FUMAGALLI P, CITTERIO S
(2007) Thiol peptide level and proteomic changes in
response to cadmium toxicity in Oryza sativa L. root. ENVIRONMENTAL AND EXPERIMENTAL BOTANY Vol 59, pp.
381-392.
AIROLDI C, PALMIOLI A, D’URZO A, COLOMBO S, VANONI
M, MARTEGANI E, PERI F (2007) Glucose-derived Ras
pathway inhibitors: evidence of Ras-ligand binding and
Ras-GEF (Cdc25) interaction inhibition. CHEMBIOCHEM
2007, Vol. 8 (12), pp. 1376-1379.
AIVATIADOU E, MATTEI E, CERIANI M, TILIA L, BERRUTI G
(2007) Impaired fertility and spermiogenetic disorders with
loss of cell adhesion in male mice expressing an interfering Rap1 mutant. MOL BIOL CELL Vol. 18(4), pp. 15301542.
BARBERIS M, KLIPP E, VANONI M, ALBERGHINA L (2007)
Cell size at S Phase Initiation: an Emergent Property of the
G1/S Network. PLOS COMPUT BIOL Vol. 3:e64.
BENZONI F, STEFANI F, STOLARSKI J, PICHON M, MITTA
G, GALLI P (2007) Debating phylogenetic relationships of
the scleractinian Psammocora: molecular and morphological evidences. CONTRIBUTIONS TO ZOOLOGY Vol 76 (1),
pp. 33-52.
BERTINI L, BRUSCHI M, DE GIOIA L, FANTUCCI P (2007)
Structure and energetics of Fe2(CO)(8) singlet and triplet
electronic states. JOURNAL OF PHYSICAL CHEMISTRY A,
Vol. 111, pp. 12152-12162.
BERTINI L, CARGNONI F, GATTI C (2007) Chemical insight
into electron density and wave functions: software developments and applications to crystals, molecular complexes
and materials science. THEORETICAL CHEMISTRY
ACCOUNTS Vol. 117, pp. 847-884.
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BORST J, RICCIARDI-CASTAGNOLI P (2007) Shaping
immunity in healthy and diseased tissues. EUR J IMMUNOL. Vol. 37(8), pp.2055-2058.
BOVOLENTA S, FOTI M, LOHMER S, CORAZZA S (2007)
+
Development of a Ca(2 )-activated photoprotein, Photina, and
its application to high-throughput screening. J BIOMOL
SCREEN. Vol. 12(5), pp. 694-704.
BRANDUARDI P, FOSSATI T, SAUER M, PAGANI R, MATTANOVICH D, PORRO D (2007) Biosynthesis of Vitamin C by
Yeast Leads to Increased Stress Resistance. PLoS ONE Vol.
2(10), e1092.
BRIANI F, DEL FAVERO M, CAPIZZUTO R, CONSONNI C,
ZANGROSSI S, GRECO C, DE GIOIA L, TORTORA P, DEHO G
(2007) Genetic analysis of polynucleotide phosphorylase
structure and functions, BIOCHIMIE Vol. 89, pp. 145-157.
CAMATTARI A, BIANCHI MM, BRANDUARDI P, PORRO D,
BRAMBILLA L (2007) Induction by hypoxia of heterologous
proteins production with the KIPDC1 promoter in yeasts.
APPL. ENVIRON. MICROBIOL. Vol. 73(3), pp. 922-929.
CARDINALE S, CISTERNA B, BONETTI P, ARINGHIERI C,
BIGGIOGERA M, BARABINO SML (2007) Subnuclear localization and dynamics of the pre-mRNA 3’ end processing
factor CF Im68. MOL. BIOL. CELL Vol. 18, pp. 1282-1292.
CAREY J, BRYNDA J, WOLFOVA J, GRANDORI R,
GUSTAVSSON T, ETTRICH R, SMATANOVA IK (2007)
WrbA bridges bacterial flavodoxins and eukaryotic
NAD(P)H: quinone oxidoreductases. PROTEIN SCI. Vol.
16, pp. 2301-2305.
CERIANI M, SCANDIUZZI C, AMIGONI L, TISI R, BERRUTI
G, MARTEGANI E (2007) Functional analysis of RalgPS2, a
murine guanine nucleotide exchange factor for RalA
GTPase. EXP CELL RES. Vol. 313(11), pp. 2293-2307.
CHIROLI E, ROSSIO V, LUCCHINI G, PIATTI S (2007) The
budding yeast PP2ACdc55 protein phosphatase prevents the
onset of anaphase in response to morphogenetic defects. J.
CELL BIOL. Vol. 177, pp. 599-611.
CIPOLLA L, REIS FERNANDES M, GREGORI M, AIROLDI C,
NICOTRA F (2007) Synthesis and biological evaluation of a
small library of nojirimycin derived bicyclic iminosugars.
CARBOHYDR. RES. Vol. 342, pp. 1813-1830.
CIPOLLINA C, VAI M, PORRO D, HATZIS C (2007) Towards
understanding of the complex structure of growing yeast
populations. J. BIOTECHNOL. Vol. 128(2), pp.393-402.
CLEMENT H, ODELL G, ZAMUDIO FZ, REDAELLI E,
WANKE E, ALAGON A, POSSANI LD. (2007) Isolation and
characterization of a novel toxin from the venom of the spider Grammostola rosea that blocks sodium channels.
TOXICON Vol. 50, pp. 65-74.
COLOSIMO E, GAMBARDELLA A, MANTEGAZZA M, LABATE A, RUSCONI R, SCHIAVON E, ANNESI F, RESTANO CASSULINI R, CARRIDEO S, CHIFARI R, CANEVINI MP, CANGER R, FRANCESCHETTI S, ANNESI G, WANKE E, QUATTRONE A (2007) Electroclinical Features of a Family with
Simple Febrile Seizures and Temporal Lobe Epilepsy
Associated with SCN1A Loss-of-Function Mutation. EPILEPSIA Vol. 48, pp. 1691-1696.
COMBI R GRIONI D, TENCHINI ML, BERTOLINI M, TREDICI G, DALPRA L (2007) Gene symbol: SCN1A. In Novel
human pathological mutations /Human Genet./ Vol.
120, 911.
COMELLI F, GIAGNONI G, BETTONI I, COLLEONI M, COSTA
B (2007) The inhibition of monoacylglycerol lipase by
URB602 showed an anti-inflammatory and anti-nociceptive
effect in a murine model of acute inflammation. BRITISH
JOURNAL OF PHARMACOLOGY Vol. 152, pp. 787-794.
CONSONNI R, AROSIO I, RECCA T, FUSI P, ZETTA L (2007)
Structural determinants responsible for the thermostability of Sso7d and its single point mutants. PROTEINS Vol. 67
(3), pp. 766-775.
CONTRAN N, CERANA R, CROSTI P, MALERBA M (2007)
Cyclosporin A inhibits the fusicoccin-induced cytochrome c
release-mediated programmed cell death of sycamore
cells. PROTOPLASMA Vol 231, pp. 193-199.
CONTRAN N, PAOLETTI E (2007) Visible foliar injury and
physiological responses to ozone in italian provenances of
Fraxinus excelsior and F. ornus. THE SCIENTIFIC WORLD
JOURNAL Vol 7(S1), pp. 90–97.
COSTA B (2007). On the pharmacological properties of
Delta9-tetrahydrocannabinol (THC). CHEMISTRY & BIODIVERSITY Vol. 4, pp. 1664-1677.
COSTA B (2007). Rimonabant: more than an anti-obesity
drug? BRITISH JOURNAL OF PHARMACOLOGY Vol. 150,
pp. 535-537.
COSTA B, TROVATO AE, COMELLI F, GIAGNONI G, COLLEONI M (2007) The non-psychoactive cannabis constituent cannabidiol is an orally effective therapeutic agent in
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rat chronic inflammatory and neuropathic pain. EUROPEAN JOURNAL OF PHARMACOLOGY Vol. 556, pp. 75-83.
DE BAKKER JM, ZAZA A (2007). Special issue on biopacemaking: clinically attractive, scientifically a challenge. MED
BIOL ENG COMPUT Vol. 45(2), pp. 115-118.
DE FILIPPIS L, LAMORTE G, SNYDER EY, MALGAROLI
A, VESCOVI AL (2007). A novel, immortal, and multipotent human neural stem cell line generatine functional
neurons and oligodendrocytes. STEM CELLS. Vol 25(9),
pp 2312-2321.
DE MATTIA F, IMAZIO S, GRASSI F, LOVICU G, TARDAGUILA J, MAITTI C, SCIENZA A, LABRA M (2007) Genetic characterization of Sardinia grapevine cultivars by SSR markers analysis. INTERNATIONAL JOURNAL OF VIGNE ET
VIN SCIENCES Vol 41, pp. 175-184.
DOULATY BANEH H, GRASSI F, MOHAMMADI A, NAZEMIEH A, DE MATTIA F, IMAZIO S, LABRA M (2007) The use
of AFLP and morphological markers to study Iranian grapevine germplasm to avoid genetic erosion. JOURNAL OF
HORTICULTURAL SCIENCE AND BIOTECHNOLOGY Vol 82,
pp. 745-752.
DOULATY BANEH H, MOHAMMADI S.A., LABRA M, NAZAMIEH A, DE MATTIA F, MARDI M (2007) Chloroplast microsatellites markers to assess genetic diversity in wild and
cultivated grapevine of Iran. PAKISTAN JOURNAL OF BIOLOGICAL SCIENCES Vol 10, pp. 1855-1859.
EILERS G, SCHWARTZ L, STEIN M, ZAMPELLA G, DE GIOIA
L, OTT S, LOMOTH R (2007) Ligand versus metal protonation of an iron hydrogenase active site mimic. CHEMISTRYA EUROPEAN JOURNAL Vol.13, pp.7075-7084.
ESTRADA G, GARCIA BI, SCHIAVON E, ORTIZ E, CESTELE
S, WANKE E, POSSANI LD, CORZO G (2007) Four disulfidebridged scorpion beta neurotoxin CssII: heterologous
expression and proper folding in vitro. BIOCHEM. BIOPHYS.
A. GEN. SUB. Vol. 1170, pp. 1161-1168.
FERRARI D, VESCOVI AL, BOTTAI D (2007) The stem cells
as a potential treatment for neurodegeneration. METHODS
MOL BIOL. Vol. 399, pp.199-213.
FILIPPONI D, HOBBS RM, OTTOLENGHI S., ROSSI P, JANNINI EA, PANDOLFI PP, DOLCI S (2007) Repression of kit
expression by Plzf in germ cells. MOLECULAR AND CELLULAR BIOLOGY [CD-ROM]. Vol. 27., pp. 6770-6781.
FORONI C, GALLI R, CIPELLETTI B, CAUMO A, ALBERTI S,
FIOCCO R, VESCOVI A (2007) Resilience to transformation
and inherent genetic and functional stability of adult neural
stem cells ex vivo. CANCER RES. Vol. 67(8), 3725-3733.
FOTI M, RICCIARDI-CASTAGNOLI P, GRANUCCI F (2007)
Gene expression profiling of dendritic cells by microarray.
METHODS MOL BIOL. Vol. 380, pp. 215-224.
GALLI P, BENZONI F, STRONA G, STEFANI F, KRITSKY D C
(2007) Monogenoidean parasites of fishes associated with
coral reefs in the Ras Mohammed National Park, Egypt:
preliminary results. HELMINTHOLOGIA Vol. 44, pp. 76-79.
GALLI P, STRONA G, BENZONI F, CROSA G, STEFANI F
(2007) Monogenoids From Freshwater Fish In Italy, With
Comments On Alien Species. COMPARATIVE PARASITOLOGY Vol. 74(2), pp. 264–272.
GALLI P, STRONA G, VILLA AM, BENZONI F, STEFANI F,
DOGLIA S M, KRITSKY DC (2007) Two-dimensional versus
three-dimensional morphometry of monogenoidean sclerites. INTERNATIONAL JOURNAL FOR PARASITOLOGY. Vol.
37, pp. 449-456.
GELAIN F, LOMANDER A, VESCOVI AL, ZHANG S (2007)
Systematic studies of a self-assembling peptide nanofiber
scaffold with other scaffolds. J NANOSCI NANOTECHNOL.
Vol. 7(2), pp. 424-434.
GIANAZZA E, WAIT R, SOZZI A, REGONDI S, SACO D,
LABRA M, AGRADI E (2007) Growth and protein profile
changes in Lepidium sativum L. plantlets exposed to cadmium. ENVIRONMENTAL AND EXPERIMENTAL BOTANY
Vol 59, pp. 179–187.
GRECO C, BRUSCHI M, DE GIOIA L, RYDE U (2007)
A QM/MM investigation of activation and catalytic mechanism of Fe-only hydrogenases. INORGANIC CHEMISTRY
Vol. 46, pp. 5911-5921.
GRECO C, BRUSCHI M, FANTUCCI P, DE GIOIA L (2007)
Influence of a large sigma-donor ligand on structural and
catalytic properties of di-iron compounds related to the
active site of Fe-hydrogenase - A DFT investigation. EUROPEAN JOURNAL OF INORGANIC CHEMISTRY Vol.13,
pp.1835-1843.
GRECO C, BRUSCHI M, HEIMDAL J, FANTUCCI P, DE GIOIA
L, RYDE U (2007) Structural insights into the active-ready
form of [FeFe]-hydrogenase and mechanistic details of its
inhibition by carbon monoxide. INORGANIC CHEMISTRY
Vol. 46, pp. 7256-7258.
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GRECO C, ZAMPELLA G, BERTINI L, BRUSCHI M, FANTUCCI P, DE GIOIA L (2007) Insights into the mechanism of
electrocatalytic hydrogen evolution mediated by
Fe2(S2C3H6)(CO)6: the simplest functional model of the
Fe-hydrogenase active site. INORGANIC CHEMISTRY Vol.
46, pp. 108-116.
INVERNIZZI G, GRANDORI R (2007) Detection of the equilibrium folding intermediate of beta-lactoglobulin in the presence of trifluoroethanol by mass spectrometry. RAPID
COMMUN. MASS SPECTROM. Vol. 21, pp. 1049-1052.
INVERNIZZI G, NATALELLO A, SAMALIKOVA M, GRANDORI R (2007) Protein-protein and protein-ligand interactions
studied by electrospray-ionization mass spectrometry.
PROTEIN AND PEPTIDE LETTERS Vol. 14, pp. 894-902.
JUSTICE AK, ZAMPELLA G, DE GIOIA L, RAUCHFUSS T B,
VAN DER VLUGT J I, WILSON, (2007) Chelate control of diiron(I) dithiolates relevant to the Fe-only hydrogenase active
site. INORGANIC CHEMISTRY Vol. 46, pp. 1655-1664.
JUSTICE AK, ZAMPELLA G, DE GIOIA L, RAUCHFUSS T B
(2007) Lewis vs. Bronsted-basicities of diiron dithiolates:
spectroscopic detection of the "rotated structure'' and
remarkable effects of ethane- vs. propanedithiolate. CHEMICAL COMMUNICATIONS Vol.20, pp.2019-2021.
KRITSKY C D, GALLI P, TINGBAO Y (2007) Dactylogyrids
(Monogenoidea) Parasitizing the Gills of Spinefoots
(Teleostei, Siganidae): Revision of Tetrancistrum Goto
and Kikuchi, 1917, with descriptions of T. strophosolenum and T. yamagutii sp. n. from Siganus spp. from the
Red Sea and Celebes. JOURNAL NATURAL HISTORY Vol.
41, pp. 1513-1551.
SGORBATI S, AIROLDI R, CITTERIO S (2007) Toxic and genotoxic effect of potassium dichromate in Pseudokirchneriella
subcapitata detected by microscopic and AFLP analysis.
AQUATIC BOTANY Vol 86, pp. 229-235.
LANAVE C, COLANGELO AM, SACCONE C, ALBERGHINA L
(2007) Molecular evolution of the neurotrophin family
members and their Trk receptors. GENE Vol. 394, pp. 1-12.
LOPEZ-VERA E, AGUILAR MB, SCHIAVON E, MARINZI C,
ORTIZ E, RESTANO CASSULINI R, BATISTA CVF, POSSANO
LD, HEIMER DE LA COTEIRA EP, PERI F, BECERRIL B,
WANKE E (2007) Novel alpha conotoxin from Conus spurius and the alpha conotoxin EI share high-affinity potentiation and low-affinity inhibition of nicotinic acetylcoline
receptors”, FEBS JOURNAL Vol. 274, pp. 3972-3985.
LOTTERSBERGER F, PANZA A, LUCCHINI G, LONGHESE
M.P (2007) Functional and physical interactions between
yeast 14-3-3 proteins, acetyltransferases, and deacetylases
in response to DNA replication perturbations. MOLECULAR
AND CELLULAR BIOLOGY. Vol. 27, pp. 3266-3281.
MALFATTO G, ZAZA A, FACCHINI M (2007) Different
effects of antiarrhythmic drugs on the rate-dependency of
QT interval: a study with amiodarone and flecainide. J CARDIOCASC PHARMACOL Vol. 50(5), pp. 535-540.
MANTIERO D, CLERICI M, LUCCHINI G, LONGHESE MP
(2007) Dual role for Saccharomyces cerevisiae Tel1 in the
checkpoint response to double-strand breaks. EMBO
REPORTS. Vol. 8, pp. 380-387.
MARZI I, D'AMICO M, BIAGIOTTI T, GIUNTI S, CARBONE
MV, FREDDUCCI D, WANKE E, OLIVOTTO M (2007) Purging
of the neuroblastoma stem cell compartment and tumor
regression on exposure to hypoxia or cytotoxic treatment.
CANCER RES. Vol. 67, pp. 2402-2407.
KRITSKY D C, GALLI P, TINGBAO Y (2007) Dactylogyrids
(Monogenoidea) Parasitizing the Gills of Spinefoots
(Teleostei, Siganidae): Proposal of Glyphidohaptor n.
gen., with Description of Two New Species from the Great
Barrier Reef, Australia, and Redescription of G. plectocirra (Paperna, 1972) comb. n. from Ras Mohammed
National Park, Egypt. JOURNAL OF PARASITOLOGY Vol.
93, pp.39-46.
MATUS-ORTEGA ME, REGONESI ME, PINA-ESCOBEDO A,
TORTORA P, DEHÒ G, GARCIA-MENA J (2007) The KH and
S1 domains of Escherichia coli polynucleotide phosphorylase
are necessary for autoregulation and growth at low temperature. BIOCHIM. BIOPHYS. ACTA. Vol. 1769, pp. 194-203.
KRITSKY D C. GALLI P (2007) Dactylogyrids (Monogenoidea)
Parasitizing the Gills of Spinefoots (Teleostei: Siganidae):
Revision of Pseudohaliotrema, with Redescriptions of P.
sphincteroporus and P. molnari from the Great Barrier
Reef, Australia. COMPARATIVE PARASITOLOGY Vol. 74(1),
pp. 9–22.
MONZANI E, FACCHETTI F, GALMOZZI E, CORSINI E,
BENETTI A, CAVAZZIN Z, GRITTI A, PICCININI A, PORRO D,
SANTINAMI M, INVERNICI G, PARATI E, ALESSANDRI G,
LA PORTA CA (2007) Melanoma contains CD133 and
ABCG2 positive cells with enhanced tumourigenic potential. EUR J CANCER Vol. 43(5), pp. 935-46.
LABRA M, BERNASCONI M, GRASSI F, DE MATTIA F,
MORO G, BONATI L, BRUSCHI M, COSENTINO U, DE GIOIA
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L, FANTUCCI PC, PANDINI A, PAPALEO E, PITEA D, SARACINO GAA, ZAMPELLA G (2007) Computational approaches
to shed light on molecular mechanisms in biological processes. THEORETICAL CHEMISTRY ACCOUNTS Vol.117,
pp.723-741.
NATALELLO A, DOGLIA SM, CAREY J, GRANDORI R (2007)
Role of flavin mononucleotide in thermostability and oligomerization of Escherichia coli stress-defense protein WrbA.
BIOCHEMISTRY Vol. 46, pp. 543-553.
NICOLIS SK (2007) Cancer stem cells and "stemness"
genes in neuro-oncology. NEUROBIOL DIS. Vol. 25,
pp.217-29.
NICOTRA F, CIPOLLA L, PERI F, LA FERLA B, REDAELLI C
(2007) Chemoselective Neoglycosylation. ADV. CARB.
CHEM. BIOCHEM. Vol. 61, pp. 353-398.
PAIARDI C, BELOTTI F, COLOMBO S, TISI R, MARTEGANI E
(2007) The large N-terminal domain of Cdc25 protein of the
yeast S. cerevisiae is required for glucose-induced Ras2
activation. FEMS YEAST RES Vol. 7(8), pp. 1270-1275.
PAOLETTI E, CONTRAN N, MANNING WJ, TAGLIAFERRO F
(2007) Ethylenediurea (EDU) affects the growth of ozonesensitive and tolerant ash (Fraxinus excelsior) trees under
ambient O3 conditions. THE SCIENTIFIC WORLD JOURNAL
Vol 7(S1), pp. 128–133.
PAPALEO E, OLUFSEN M, DE GIOIA L, BRANDSDAL B
(2007) Optimization of electrostatics as a strategy for coldadaptation: A case study of cold- and warm-active elastases. JOURNAL OF MOLECULAR GRAPHICS & MODELLING
Vol. 26, pp.93-103.
PAVELKA N, FOURNIER ML, SWANSON SK, PELIZZOLA M,
RICCIARDI-CASTAGNOLI P, FLORENS L, WASHBURN MP
(2007) Statistical similarities between transcriptomics and
quantitative shotgun proteomics data. MOL CELL PROTEOMICS 2007 Nov 19.
PERI F, GRANUCCI F, COSTA B, ZANONI I, MARINZI C,
NICOTRA F (2007) Inhibition of Lipid A stimulated activation
of human dendritic cells and macrophages by amino and
hydroxylamino monosaccharides. ANGEWANDTE CHEMIE.
INTERNATIONAL EDITION. Vol. 46, pp. 3308-3312.
PICCIRILLO SG, VESCOVI AL (2007) Brain tumour stem
cells: possibilities of new therapeutic strategies. EXPERT
OPIN BIOL THER. Vol. 7(8): pp. 1129-35.
PICHON M, BENZONI F (2007) Taxonomic re-appraisal of
zooxanthellate Scleractinian Corals in the Maldive
Archipelago. ZOOTAXA Vol. 1441, pp. 21-33.
PILLOZZI S, BRIZZI MF, BERNABEI PA, BARTOLOZZI B,
CAPORALE R, BASILE V, BODDI V, PEGORARO L, BECCHETTI A, ARCANGELI A (2007) VEGFR-1 (FLT-1), ß1 integrin and hERG K+ channel form a macromolecular signaling complex in acute myeloid leukemia: role in cell migration and clinical outcome. BLOOD Vol 110, pp. 1238-1250.
PODLIPNIK Â, VELTER I, LA FERLA B, MARCOU G, BELVISI L, NICOTRA F, BERNARDI A (2007) First round of focused library for cholera toxin inhibitor search. CARBOHYDR.
RES. Vol. 342, pp. 1651-1660.
PROSPERI D, MARASSO C, TORTORA P, MONTI D, BELLINI T (2007) Avidin decorated core-shell nanoparticles for
biorecognition studies by elastic light scattering. CHEMBIOCHEM. Vol. 8, pp. 1021-1028.
QUERIN L, SANVITO R, MAGNI F, BUSTI S, VANDORSSELAER A, ALBERGHINA L, VANONI M (2007) Proteomic analysis of a nutritional shift-up in S. cerevisiae identifies
Gvp36 as a BAR-containing protein involved in vesicular
traffic and nutritional adaptation J BIOL CHEM Epub (2007)
Dec 21.
RICCHELLI F, FUSI P, TORTORA P, VALTORTA M, RIVA M,
TOGNON G, CHIEREGATO K, BOLOGNIN S, ZATTA P (2007)
Destabilization of non-pathological variants of ataxin-3 by
metal ions results in aggregation/fibrillogenesis. INT. J.
BIOCHEM. CELL BIOL. Vol. 39, pp. 966-977.
RIZZI CT, CARVALHO-DE-SOUZA JL, SCHIAVON E, CASSOLA AC, WANKE E, TRONCONE LR (2007) Crotamine inhibits preferentially fast-twitching muscles but is inactive on
sodium channels. TOXICON Vol. 50, pp. 553-562.
RUBOLINI D, AMBROSINI R, CAFFI M, BRICHETTI P,
ARMIRAGLIO S, SAINO N (2007). Long-term trends in first
arrival and first egg laying dates of some migrant and resident bird species in northern Italy. INTERNATIONAL JOURNAL OF BIOMETEOROLOGY Vol. 51, pp. 553-563.
RUSCONI R, SCALMANI P, RESTANO CASSULINI R, GIUNTI G, GAMBARDELLA A, FRANCESCHETTI S, ANNESI G,
WANKE E, MANTEGAZZA M (2007) Modulatory proteins
can rescue a trafficking defective epileptogenic Nav1.1
(SCN1A) Na+ channel mutant. J. NEUROSCI. Vol. 27, pp.
11037–11046.
SAINO N, AMBROSINI R (2007). Climatic connectivity between Africa and Europe may serve as a basis for phenotypic
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adjustment of migration schedule of trans-Saharan migratory birds. GLOBAL CHANGE BIOLOGY Vol. 14, pp. 1-14.
and EFTu. AMERICAN JOURNAL OF HUMAN GENETICS
Vol. 80 pp.44-58.
SCHNEIDER C, ZAMPELLA G, GRECO C, PECORARO V L,
DE GIOIA L (2007) Mechanistic analysis of nucleophilic substrates oxidation by functional models of vanadium-dependent haloperoxidases: A density functional theory study.
EUROPEAN JOURNAL OF INORGANIC CHEMISTRY Vol. 4,
pp. 515-523.
VELTER AI, POLITI M, PODLIPNIK C, NICOTRA F (2007)
Natural and Synthetic Cholera Toxin Antagonists, MINIREV. MED. CHEM Vol. 7, pp. 159-170
SPERANDEO P, CESCUTTI R, VILLA R, DI BENEDETTO C.
CANDIA D, POLISSI A (2007) Characterization of lptA and
lptB, two essential genes implicated in lipopolysaccharide
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ZAZA A. (2007) Fisiologia. Molecole, cellule e sistemi. A
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2007
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PORRO D, BRANDUARDI P, MATTANOVICH D, SAUER M.
Increase in stress tolerance with ascorbic acid during fermentation. US2007141687 Filing Date: 21/06/07
VALLI M, BRANDUARDI P, ALBERGHINA L, PORRO D.
Producing a protein useful as pharmaceuticals, e.g. medicine and vaccine, or in food or paper production, comprises expressing and secreting a protein expressed by
Zygosaccharomyces bailii strain. DE10252245 Filing Date:
2007-05-27
PORRO D, SAUER M. Ascorbic acid production from yeasts.
DE60118200T T2 Filing Date: 12/04/07 ; DE60127470D
Filing Date: 03/05/07; AT357533 Filing Date: 15/04/07;
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Date: 29/11/07
VALLI M, SAUER M, PORRO D, BRANDUARDI P, MATTANOVICH D. Strains for the Production of Organic Acids.
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SAUER M, PORRO D, BRANDUARDI P, MATTANOVICH D.
Improved strains for the production of organic acids.
WO2007038130 A3 Filing Date: 31/05/07; WO2007038130
A2 Filing Date: 05/04/07
BRANDUARDI P, PORRO D, VALLI M, ALBERGHINA L.
Process for expression and secretion of proteins by the
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