Direzione e uffici C.I.B.

Direzione CIB:
Prof. Claudio Schneider
Email: claudio.schneider@Lncib.it

Segreteria CIB:
Prof. Roberto Gambari
Email: roberto.gambari@unife.it

SEGRETERIA ORGANIZZATIVA:
Elisabetta Lambertini
Tel: 0532/974451
Fax: 0532/974484
E-mail: lmblbt@unife.it

AMMINISTRAZIONE:
Vanessa Florit
Area di Ricerca
Padriciano, 99 - 34012 Trieste
Tel: 040/398979
Fax: 040/398990
E-mail: cib@lncib.it

Posta certificata C.I.B.:
cib@poste-certificate.it

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Finotti Paola Stampa
RESPONSABILE DELLA U. O.

Cognome e Nome Finotti Paola
Qualifica RICERCATORE CONFERMATO
Facoltà MEDICINA-CHIRURGIA
Dipartimento FARMACOLOGIA E ANESTESIOLOGIA
Settore Scientifico Disciplinare BIO/14
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PERSONALE STRUTTURATO

Cognome e Nome
Brunati Anna Maria
Qualifica
PA
Dipartimento Chimica Biologica
Ente di appartenenza Università di Padova
Cognome e Nome
Caparrotta Laura
Qualifica
PA
Dipartimento Farmacologia e Anestesiologia
Ente di appartenenza Università di Padova
Cognome e Nome
Facchinetti Antonella
Qualifica
Tecnico
Dipartimento
Scienze Oncologiche e Chirurgiche
Ente di appartenenza
Università di Padova
Cognome e Nome
Finotti Paola
Qualifica
RU
Dipartimento
Farmacologia e Anestesiologia
Ente di appartenenza
Università di Padova
Cognome e Nome
Montopoli Monica
Qualifica
Dottorando
Dipartimento Farmacologia e Anestesiologia
Ente di appartenenza
Università di Padova
Cognome e Nome
Pagetta Andrea
Qualifica
Tecnico
Dipartimento Farmacologia e Anestesiologia
Ente di appartenenza
Università di Padova
Cognome e Nome
Tibaldi Elena
Qualifica
Dottorando
Dipartimento
Chimica Biologica
Ente di appartenenza
Università di Padova
Cognome e Nome Tramentozzi Elisa
Qualifica Dottorando
Dipartimento
Farmacologia e Anestesiologia
Ente di appartenenza
Università di Padova
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza

PERSONALE NON STRUTTURATO

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Qualifica
Dipartimento
Ente di appartenenza
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza
Cognome e Nome
Qualifica
Dipartimento
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Cognome e Nome
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Dipartimento
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Dipartimento
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LINEE DI RICERCA

The research project is designed to identify the molecule(s) and mechanisms involved in the development of vascular complications which occur in diabetes at a higher frequency and severity compared with the normal population. To this aim, an extensive proteomic analysis on plasma proteins, from both control and type 1 diabetic subjects, is performed to identify and characterize the structure and function of molecules which best match requisites of markers of biological relevance. Studies are mainly oriented on proteins which display serine protease activity, although they do not belong to the class of classical serine proteases. In particular, attention is centered on some Heat Shock Proteins (HSPs) whose plasma concentration is observed to be abnormally increased in diabetic subjects, and on IgG antibodies (Abs) found complexed with HSPs. Since some HSPs, such as Grp94, bear serine-like protease activity, experiments are set up to investigate in depth both biochemical and immunological properties of these molecules, to shed light on the complex mutual influence the proteolytic activity may have on stimulation of the immune system. The research thus also deals with structural and functional characterization of Abs developed against HSPs, with the specific aim to investigate the mechanism by which Abs become catalytic, and to establish whether immune complexes of catalytic Abs drive any angiogenic effects. To this purpose, experiments are set up to study phenotypic changes of, and proliferative effects on human endothelial cells, and to identify the intracellular pathway involved in cell activation, proliferation and differentiation. These experiments include, among others, the detection of phosphorylated mitogen-activated protein kinase in both absence and presence of inhibitors acting on specific metabolic steps, also employing as positive controls classical serine proteases whose molecular mechanism is already known. In addition, HSPs of interest are expressed in cell cultures of E. coli and tested, both singularly and in complexes with human IgG, in experiments of comparison with plasma-purified HSPs and IgG Abs. The discovery of plasma molecules which are valuable vascular risk markers can be translated into diagnostics and therapeutics for early detection and cure of vascular diseases.


TECNOLOGIE IN POSSESSO DELL'U. O.

  • Protein expression and purification from E. coli
  • Protein purification from human plasma (ion-exchange, gel-filtration, affinity chromatography, FPLC, HPLC)
  • Immunoblotting, Immuno-precipitation, Two-dimensional electrophoresis
  • Spectral analysis: UV and fluorescence spectrometry, mass analysis (external service with collaboration)
  • Measurements of enzyme activity; zymography
  • Assays for phosphoprotein detection and protein kinase activities; cell signalling and MAP technologies
  • Cell cultures; tests of viability, proliferation and cytotoxicity; phenotypic screen and molecular target-based screen; flow cytometry and immunofluorescence

STRUMENTAZIONE

Denominazione
Struttura ove la strumentazione è allocata
Responsabile della strumentazione
Nikon C1 Confocal microscope (Software Nikon EZ-C1 2.10)
Dipartimento di Farmacologia
Dott. Daga Andrea
Cytofluorometer XL Coultar (2)
Dip. Farmacologia e Dip. Scienze Oncologiche e Chirurgiche
Dott.ssa Monopoli Monica e Dott.ssa Antonella Facchinetti
Pharmacia FPLC System
Dipartimento di Farmacologia
Dott. Pagetta Andrea
Shimadzu UV-visible recording Spectrophotometer UV-1601
Dipartimento di Farmacologia
Nessun responsabile specifico
Beckman Coulter Avanti J-20XP centrifuge
Dipartimento di Farmacologia
Dott. Pagetta Andrea
Perkin Elmer Luminescence Spectrofluorometer LS-50
Dipartimento di Farmacologia
Nessun responsabile specifico

PUBBLICAZIONI

1. Brunati AM. Contri A. Muenchbach M. James P. Marin O. Pinna LA. GRP94 (endoplasmin) co-purifies with and is phosphorylated by Golgi apparatus casein kinase. FEBS Lett. 2000;471(2-3):151-5.
1. Pagetta A. Folda A. Brunati AM. Finotti P. Identification and purification from the plasma of Type 1 diabetic subjects of a proteolytically active Grp94. Evidence that Grp94 is entirely responsible for plasma proteolytic activity. Diabetologia. 2003;46(7):996-1006.
3. Deana R. Turetta L. Donella-Deana A. Dona M. Brunati AM. De Michiel L. Garbisa S. Green tea epigallocatechin-3-gallate inhibits platelet signalling pathways triggered by both proteolytic and non-proteolytic agonists. Thromb Haemost. 2003;89(5):866-74.
4. Finotti P. Pagetta A. A heat shock protein70 fusion protein with alpha1-antitrypsin in plasma of type 1 diabetic subjects. Biochem Biophys Res Commun. 2004;315(2):297-305.
5. Finotti P. Pagetta A. Ashton T. The oxidative mechanism of heparin interferes with radical production by glucose and reduces the degree of glycooxidative modifications on human serum albumin. Eur J Biochem. 2001;268(8):2193-200.
6. Ragazzi E. Costa CV. Caparrotta L. Biasiolo M. Bertazzo A. Allegri G. Enzyme activities along the tryptophan-nicotinic acid pathway in alloxan diabetic rabbits. Biochim Biophys Acta. 2002;1571(1):9-17.
8. Finotti P. The role played by serine proteases in the development and worsening of vascular complications in Type 1 diabetes mellitus. Invited Review. Current Diabetes Review. 2006; in press.
9. Facchinetti A. Santa SD. Mezzalira S. Rosato A. Biasi G. A large number of T lymphocytes recognize Moloney-murine leukemia virus-induced antigens, but a few mediate long-lasting tumor immunosurveillance. J Immunol. 2005;174(9):5398-406.
10. A. Pagetta, E. Tramentozzi, L. Corbetti, M. Frasson, A. M. Brunati and P. Finotti. Characterization of immune complexes of idiotypic catalytic and anti-idiotypic inhibitory antibodies in plasma of Type 1 diabetic subjects. Mol. Immunol. 44: 2870-2883, 2007.

DOTTORATI DI RICERCA

Componente U.O. Dottorato di Ricerca Coordinatore Sede
Finotti Paola
International Research Doctorate on Molecular and Cellular Pharmacology
Prof. Sisto Luciani
Padova Innsbruck Freiburg
Caparrotta Laura
International Research Doctorate on Molecular and Cellular Pharmacology
Prof. Sisto Luciani
Padova Innsbruck Freiburg

CONGRESSI C.I.B.

Congressi Partecipazione
E. Tramentozzi, A. Pagetta and P. Finotti. Both a
Si