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

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

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

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De Luca Giancarlo Stampa
RESPONSABILE DELLA U. O.

Cognome e Nome De Luca Giancarlo
Qualifica
full professor
Facoltà
School of Medicine
Dipartimento Deparetment of Biomedical Experimental and Clinical Sciences (DSBSC) University of Insubria Varese
Settore Scientifico Disciplinare
Bio/10
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PERSONALE STRUTTURATO

Cognome e Nome Karousou Evgenia
Qualifica
RU
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome
Moretto Paola
Qualifica
Technician
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome
Pallotti Francesco
Qualifica
PA
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome
Passi Alberto
Qualifica
PO
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome Vigetti Davide
Qualifica
RU
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome
Viola Manuela
Qualifica
RU
Dipartimento
DSBSC
Ente di appartenenza University of Insubria Varese
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza

PERSONALE NON STRUTTURATO

Cognome e Nome
Bartolini Barbara
Qualifica
Post doc
Dipartimento DSBSC
Ente di appartenenza
University of Insubria Varese
Cognome e Nome Clerici Moira
Qualifica PhD student
Dipartimento DSBSC
Ente di appartenenza
University of Insubria Varese
Cognome e Nome
Genasetti Anna
Qualifica Post doc
Dipartimento DSBSC
Ente di appartenenza
University of Insubria Varese
Cognome e Nome
Rizzi Manuela
Qualifica PhD student
Dipartimento DSBSC
Ente di appartenenza
University of Insubria Varese
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza

LINEE DI RICERCA

The main interest of the De Luca’s group is the glycosaminoglycan (GAG) metabolism. GAGs are linear sulphated polysaccharides containing uronic acid and hexosamine residues. These molecules are very common in nature and linked to a protein core constitute the proteoglycan family. Hyaluronan (HA) is a special GAG without core protein and completely unsulphated. The aim of our work is the study of the mechanisms driving the GAG synthesis. In order to achieve this target we use a biochemical approach integrated with molecular biology techniques and morphology. To address the GAG chemistry we set up a new electrophoretic method based on the carbohydrate fluorescence (FACE). This method is very simple, fast and able to detect few pmoles of samples. Our preliminary data indicate that over expression in cells of UGDH, the enzyme involved in the synthesis of UDP-glucuronic acid (UDPGlcUA), the UDP uronic precursors of all GAGs, induced an increase in the cells of this precursor and consequently an increase of HA synthesis. On the other hand the silencing of the same enzyme by siRNA induced in the cells the opposite effect, confirming that the availability of UDPGlcUA controls the HA synthesis. In order to quantify the UDP sugars precursors in the cells, we developed a method using zone capillary electrophoresis (CZE) and HPLC able to separate and quantify all UDP sugars involved in GAG synthesis. Moreover, the altered UDP sugar concentration in the cells was able to alter the expression of the enzymes involved in the UDP sugars synthesis and even the enzymes involved in GAG synthesis. On this basis we are now elucidating the mechanisms involved in this control of enzymes of the pathway of GAG synthesis. For this purpose we also consider the post transcriptional modification of the enzymes using a proteomic approach based on bi-dimensional electrophoresis and mass spectrometry analysis coupled with a genomic approach base on the microarray chips and real time RT-PCR.


TECNOLOGIE IN POSSESSO DELL'U. O.

  • HPLC and carbohydrate
  • FACE electrophoresis for carbohydrate
  • Zone Capillary electrophoresis CZE for carbohydrate
  • Confocal Cell Fluorescence
  • Cell cultures
  • Transient transfection by nucleofection
  • Signal silencing using siRNA
  • Cell membrane impedance (ECIS).
  • Carbohydrate Mass spectrometry
  • Real time RT-PCR

STRUMENTAZIONE

Denominazione
Struttura ove la strumentazione è allocata
Responsabile della strumentazione
Real time PCR
DSBSC
Moretto
Ultracentrifuge
DSBSC
Moretto
Capillary electrophoresis
DSBSC
Passi
HPLC/FPLC with fluorescence detector
DSBSC
Viola
Nucleofector and cell cultures
DSBSC
Vigetti
Fluorescence microscopy
DSBSC
Karousou

PUBBLICAZIONI

Vigetti D., Viola M., Gornati R., Ori M., Nardi I., Passi A., De Luca G., Bernardini G. “Molecular cloning, genomic organization and developmental expression of the Xenopus laevis hyaluronan synthase 3”. (2003) In Matrix biology, 22, pagg 511-517.

Passi A., Sadeghi P., Kawamura H., Anand S., Sato N., White L.E., Hascall V.C., Maytin E.V. hyaluronan suppresses late markers of epidermal differentiation in organotypic cultures of rat keratinocytes (2004) Experimental Cell Research 296/2, 123-134

Karousou E.G., Militsopoulou M., Porta G., De Luca G., Hascall VC, Passi A. Polyacrylamide gel electrophoresis of fluorophore-labeled hyaluronan and chondroitin sulfate disaccharides. Application to the analysis in cells and tissues. (2004) Electrophoresis vol. Volume 25, Issue 17 , Pages 2919 - 2925

Luigi Raio, Antonella Cromi, Fabio Ghezzi, Alberto Passi, Eugenia Karousou, Manuela Viola, Davide Vigetti , Giancarlo De Luca and Pierfrancesco Bolis. Hyaluronan content of Wharton´s jelly in healthy and Down syndrome fetuses. (2005) Matrix biology, vol 24 (2) : 166-74.

Negrini D, Tenstad O, Passi A, Wiig H. Differentiale degradation of matrix proteoglycans and edema development in rabbit. (2006) Am J Physiol Lung Cell Mol Physiol. 290 (3): L470-L477

Vigetti D, Ori M, Viola M, Genasetti A, Karousou E, Rizzi M, Pallotti F, Nardi I, Hascall VC, De Luca G and Passi A Molecular cloning and characterization of UDP-glucose dehydrogenase from the amphibian xenopus laevis and its involvement in hyaluronan synthesis (2006) J. Biol. Chem, 281 8254-8263

Vigetti D, Moretto P, Viola M, Genasetti A, Rizzi M, Karousou E, Pallotti F, De Luca G and Passi A. Matrix metalloproteinase 2 and tissue inhibitors of metalloproteinases regulate human aortic smooth muscle cell migration during in vitro aging. (2006) FASEB J. 20, 1118-1130 (2006).

Moriondo A., Pelosi P., Passi A., Viola M., Marcozzi C., Severgnini P., Ottani V., Quaranta M., Negrini D. Proteoglycans fragmentation and respiratory mechanics in mechanically ventilated healthy rats. J Appl Physiol. 2007 Sep;103(3):747-56.

Genasetti A, Valentino ML, Carelli V, Vigetti D, Viola M, Karousou EG, Melzi d'Eril GV, De Luca G, Passi A, Pallotti F. Assessing Heteroplasmic Load in Leber's Hereditary Optic Neuropathy Mutation 3460G->A/MT-ND1 with A Real-Time PCR Quantitative Approach. J Mol Diagn. 2007 Sep;9(4):538-45. Epub 2007 Jul 25. PMID: 17652639

Vigetti D, Viola M, Karousou E, Rizzi M, Moretto P, Genasetti A, Clerici M, Hascall VC, De Luca G, Passi A. Hyaluronan-CD44-ERK1/2 regulate human aortic smooth muscle cell motility during aging. J Biol Chem. 2008 Feb 15;283(7):4448-58.

DOTTORATI DI RICERCA

Componente U.O. Dottorato di Ricerca Coordinatore Sede
De Luca
Biotecnologie
De Luca
Varese
Passi
Biotecnologie
De Luca
Varese
Vigetti
Biotecnologie
De Luca
Varese

CONGRESSI C.I.B.

Congressi Partecipazione
CNB4

CNB5

CNB6

CNB7

CNB8

CNB9
CNB10