Stefano Cotgnoli has extensive experience presenting and publishing peer-reviewed research. He has expertise in protein biochemistry, molecular biology, cell biology, and small molecule analysis. His doctoral thesis focused on the Ctr family of copper transporters and the regulation and function of Ctr2 in copper homeostasis. He has strong skills in research design, bioinformatics, and scientific techniques.

1. Cotgnoli Stefano additonal CV informaton
SELECTION OF PEER REVIEWED PRESENTATIONS AND PUBLICATIONS
Biol Trace Elem Res 2009
Trace Elements and Metallothionein in Liver and Kidney of Felis catus.
Andreani G, Cotgnoli S, Perfet B, Kismali G, Carpené E, Isani G.
Metallothioneins as clinical biomarkers in veterinary medicine.
Andreani G, Cotgnoli S(presenter), Asseta B, Ricci A, Senzolo M, Carpené E, Isani G
SEVC 2009 – Southern European Veterinary Conference (October 17‐19, 2009).
Barcelona, Spain.
Studies on the cell biology and functon of mammalian Ctr2.
Cotgnoli S (presenter), Nose Y, Kim BE and Thiele DJ.
FASEB summer Research Conference. Trace Element Micronutrients: From Model Organisms to
Humans (June 15‐20, 2008).
Snowmass Village, CO, USA.
Sci Total Environ 2008;390(1):287‐94.
Metal distributon and metallothionein in loggerhead (Careta careta) and green (Chelonia
mydas) sea turtles.
Andreani G, Santoro M, Cotgnoli S, Fabbri M, Carpené E, Isani G.
Expression and cellular localizaton of endogenous mouse Ctr2.
Cotgnoli S (presenter), Nose Y, McNaughton K, Andreani G and Thiele DJ.
Gordon Research Conference. Cell Biology of Metals (July 29 ‐ August 3, 2007).
Salve Regina University, Newport, RI, USA.
Metallothionein cDNA cloning and RT‐PCR inhibitors in Scapharca inaequivalvis.
Andreani G, Capranico G, Cotgnoli S (speaker), Isani, Monari M, Fabbri M and
Carpené E.
23th ESCPB (European Society for Comparatve Physiology and Biochemisty) Congress
(September 23‐26, 2004).
Cesenatco, Italy.

2. DOCTORAL THESIS ABSTRACT
The Ctr family is an essental part of the copper homeostasis machinery and its members share
sequence homology, structural and functonal features. Higher eukaryotes express two members
of this family Ctr1 and Ctr2.
Numerous structural and functonal studies are available for Ctr1, the only high afnity Cu(I)
transporter thus far identfed. Ctr1 holigotrimers mediate cellular copper uptake playing a crucial
role in dietary copper acquisiton and it has been demonstrated to be essental for embryonic
development.
Instead very litle is known about Ctr2, it bears structural homology to the yeast vacuolar copper
transporter, which mediates mobilizaton of vacuolar copper stores.
Recent studies using over‐expressed epitope‐tagged forms of human Ctr2 suggested a functon as
a low afnity copper transporter, which can mediate either copper uptake from the extracellular
environment or mobilizaton of lysosomal copper stores.
Using an antbody that recognizes endogenous mouse Ctr2 as confrmed by antbody‐peptde
competton assay and siRNA knockdown experiments, we studied the expression and localizaton
of endogenous mouse Ctr2 in cell culture and in mouse models in order to understand its
regulaton and functon in copper homeostasis.
By immunoblot we observed a regulaton of mCtr2 protein levels in a copper and Ctr1 dependent
way. While additon of copper to the media caused a tme dependent decrease of Ctr2 protein
expression, copper chelaton treatment had an opposite efect, similarly to what had been
previously described for Ctr1.
Interestngly in cultured cell lines derived from Ctr1 knockout embryos, Ctr2 protein levels appear
to be strongly down‐regulated and almost undetectable when compared to cell lines derived from
wild type embryos.
We then tested the expression of Ctr2 in tssues of control and Ctr1 transgenic mice.
Of partcular relevance were the results obtained in Ctr1 conditonal knockout mice, lacking Ctr1
specifcally and exclusively in the intestnal epithelium cells (IECs), and characterized by severe
copper defciency in all peripheral tssues.
In Ctr1 null IECsCtr2 expression was basically non detectable, while in peripheral tssues, and in
partcular in the kidney, Ctr2 levels were higher in conditonal knockout mice when compared to
tssues of control animals, therefore recapitulatng in vivo both the copper and Ctr1 dependent
regulaton of Ctr2 previously observed in vitro.

3. By indirect immunofuorescence we observed in several cell lines exclusive intracellular localizaton
in a perinuclear compartment and lack of co‐localizaton with lysosomal markers.
As excepton only in Ctr1 null cells we detected partal localizaton of Ctr2 in granule‐like structures
co‐localizing with lysosomal markers.
These preliminary results suggest a model in which Ctr1 and Ctr2 are co‐regulated in response to
copper, possibly by direct interacton, and lack of such interacton cause trafcking and
degradaton of Ctr2 into the lysosomes.
More data will be required to support this hypothesis and to address the functon of Ctr2 in higher
eukaryotes.
In appendix and unrelated with the subject discussed above, a LC‐mass method for analysis of algal
biotoxins belonging to the family ofPsP (Paralytc Shellfsh Poisoning) is described.
Gradient eluton on a HILIC column employing low concentratons of TFA, allowed a good
resoluton and good signals for all the PsP standards commercially available.
Preliminary tests suggest the possibility of using a weak caton exchange SPE cartridge to partally
counteract the strong matrix efect observed in mollusk samples.
SUMMARY OF RESEARCH QUALIFICATIONS AND TRAINING
Scientfc Techniques/Laboratory Experience :
• Protein Biochemistry: extracton from cells and t ssues, diferental centrifugaton,
quantfcaton, gel fltraton chromatography, atomic absorpton spectrophotometry,SDS‐
page, western blotng and immunoprecipitaton, ingel and insoluton tryptc digeston,
Maldi‐Tof analysis, Mass fngerprintng.
• Molecular biology: Primer design, DNA & RNA extracton, enzymatc digeston and ligaton,
RT‐PCR, electrophoresis.
• Cell biology: media preparaton, expansion, cryostocking, siRNA gene silencing, indirect
immunofuorescence and live cell fuorescence microscopy.
• Small molecule analysis: solid phase extracton, HPLC and LC‐mass(SIR and SRM) analysis
of hydrophilic and lipophilic compounds in biological samples.
Bioinformatcs:
• Database browsers: nucleotde (Genebank), protein (UniProt/Swiss‐Prot) and genome
(NCBI, MGI, Ensembl, Vega).
• Sequence analysis: Protein and nucleotde alignment (BLAST, ClustalW), Transmembrane
helices predicton in proteins (TMHMM), oligonucleotde analysis (OligoCalc,
OligoAnalyzer), Peptde Mass Fingerprintng Tools (Aldente, Mascot).
• Primer and RNAi designs: Primer3, PrimerQuest, siDESIGN center Dharmacon, sIR,
sirnaSfold, RNAsx.

4. Project design skills:
• Designed and developed a research project aimed to evaluate expression of
metallothioneins as potental diagnostc biomarkers in canine tumor.
• Autonomously designed an original project to study producton of biodiesel precursors
from micro‐algae and submited it for an internatonal fellowship sponsored by ENI (the
major italian oil company) and ISSNAF (The Italian Scientsts and Scholars of North America
Foundaton). Click here to read the project.
COURSES
Name: AsterDOC 2009: Science and Technology in Business.
Organizaton: Aster (Consortum among the Emilia‐Romagna Regional Government, regional
Universites and Natonal Research Centers, ENEA, CNR, INFM and the Entrepreneurial Associatons
of the region).
Place and date: CNR Area della Ricerca, Bologna (Italy) 6‐11/07/2009.
Name: Peer Review in Theory and Practce: Training course for scientfc ofcers of grantng
insttutons.
Organizaton: The European Genetcs Foundaton, Telethon Italia, the PROGEN Consortum and the
University of Bologna.
Place and date: EuroMediterranean University Centre, Ronzano (Italy), 8‐10/11/2008.
REFERENCES
Prof. Francisco Veloso
Associate Professor at the School of Business and Economics, Universidade Catolica Portuguesa
(Lisbon, Portugal)
Associate Professor at the Department of Engineering and Public Policy, Carnegie Mellon University
(Pitsburgh, U.S.A.)
Phone: +351 217214270
Email: fveloso@clsbe.lisboa.ucp.pt
Cristna Campos
Mixed Specialty BF Director
Novarts Farma ‐ Produtos Farmaceutcos S.A. (Lisbon, Portugal).
Phone: +351 21 0008720
Email: cristna.campos@novarts.com
Prof. Emilio Carpené
Full Professor at the Department of Clinical Veterinary‐Faculty of Veterinary Medicine University of
Bologna (Bologna, Italy).
Director of the Inter‐Departmental Research Center for Marine Sciences
Doctoral thesis advisor
Phone: +39 051‐2097025
Email: emilio.carpene@unibo.it