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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Arcari Paolo Stampa
RESPONSABILE DELLA U. O.

Cognome e Nome Arcari Paolo
Qualifica Professore Ordinario
Facoltà Medicina e Chirurgia, Università di Napoli Federic
Dipartimento Biochimica e Biotecnologie Mediche
Settore Scientifico Disciplinare BIO/10
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PERSONALE STRUTTURATO

Cognome e Nome Antonio Dello Russo
Qualifica PA
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Emilia Rippa
Qualifica
FT
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Emmanuele De
Qualifica
PO
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Gaetano Corso
Qualifica
PA
Dipartimento
Dipartimento di Scienze Biomediche
Ente di appartenenza Università di Foggia
Cognome e Nome Gennaro Raimo
Qualifica
PA
Dipartimento Dipartimento di Scienze e Tecnologie per l’Ambiente e il Territorio
Ente di appartenenza
Università del Molise
Cognome e Nome Maria Fiammetta Romano
Qualifica
RU
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Mariorosario Masullo
Qualifica
PA
Dipartimento Dipartimento di Scienze Farmacobiologiche
Ente di appartenenza
Universita` di Catanzaro Magna Graecia
Cognome e Nome Paolo Arcari
Qualifica
PO
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome
Rita Bisogni
Qualifica
FT
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Rosaria Ruocco
Qualifica
RU
Dipartimento
Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza

PERSONALE NON STRUTTURATO

Cognome e Nome Annalisa Lamberti
Qualifica
Contratto
Dipartimento Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Carmen Sanges
Qualifica Dottorato
Dipartimento Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Giovanna La Monica
Qualifica Dottorato
Dipartimento Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome Immacolata Ruggiero
Qualifica Dottorato
Dipartimento Dipartimento di Biochimica e Biotecnologie Mediche
Ente di appartenenza Università di Napoli Federrico II
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza
Cognome e Nome
Qualifica
Dipartimento
Ente di appartenenza

LINEE DI RICERCA

The translation elongation factors in extremophilic organisms and in eukaryotes (Prof. P. Arcari, Prof. M. Masullo, Prof. G Raimo, Dr. A. Lamberti, Dr. E. Rippa) The elongation factors SsEF-1α, SsEF-2 and SsEF-1β have been isolated from the archaeon Sulfolobus solfataricus and the corresponding genes have been isolated and sequenced. Recombinant forms of these elongation factors have been produced in E. coli. Also several mutated forms have been designed and produced in order to carry on experiments aimed to enlarge the knowledge on the structure-relationship in proteins. Recently, the 3D structure of SsEF-1α in complex with GDP has been resolved at 1.8 Ä resolution. The research project is to study the 3D structure of the mutated forms and toThe translation elongation factors in extremophilic organisms and in eukaryotes (Prof. P. Arcari, Prof. M. Masullo, Prof. G Raimo, Dr. A. Lamberti, Dr. E. Rippa) The elongation factors SsEF-1α, SsEF-2 and SsEF-1β have been isolated from the archaeon Sulfolobus solfataricus and the corresponding genes have been isolated and sequenced. Recombinant forms of these elongation factors have been produced in E. coli. Also several mutated forms have been designed and produced in order to carry on experiments aimed to enlarge the knowledge on the structure-relationship in proteins. Recently, the 3D structure of SsEF-1α in complex with GDP has been resolved at 1.8 Ä resolution. The research project is to study the 3D structure of the mutated forms and to obtain the 3D structure of SsEF-1α in complex with GTP and with the exchange factor SsEF-1β. The elongation factors EF-Tu and EF-Ts have been isolated and purified from the Antarctic microorganism Pseudoalteromonas haloplanktis (formerly Moraxella). Also in this case, the corresponding gene have been isolated and sequenced . The other elongation factor EF-G has been purified and partially characterized. The research will continue by producing mutated forms of these enzymes in order to study their biochemical properties and their resistance against physical and chemical denaturants. The aim of the research is to compare the biochemical and structural properties of the psichrophilic enzymes with those of the corresponding thermophilic and mesophilic ones. The phenylalanine-tRNA synthetase from S. solfataricus (SsFRS) has been characterized. This enzyme is very important for the formation or the complex between Phe-tRNAPhe and SsEF-1α. The enzyme is specific for tRNA from S. solfataricus. In addition, the enzyme shows a marked thermostability and resistance against chemical denaturants. The development of the project is directed toward the production of large quantities of the enzyme for 3D structural characterization. A recent project regarding the functional role of the translation elongation factor 1A in human cells has been initiated. The study is carried out by using as experimental system a human carcinoma epidermoid cell line (H1355). The involvement of EF-1A during the apoptotic process induced by interferon-a showed that EF-1A undergoes to a phosphorylation process.

DNA interferometric biosensor on porous silicon (Prof. P. Arcari, Dr. A. Lamberti) The project is focused on the construction of a DNA biosensor based on porous silicon technology. In fact, the use of porous silicon as solid support for chemical recognition gives enormous advantages because porous silicon has a very large porous surface that might allow more efficient interactions inside the porous cavity. The detection of the chemical interaction is traduced in an optical signal that is evaluated by the Fabry-Perot effect. In fact, the molecular recognition is based on the variation of the refractive index of the porous cavities that occurs if these cavities are filled with molecular masses. Porous silicon, once prepared, shows a surface terminating with Si-H bonds. The aim of the project is to try to chemically attach to the porous silicon surface linkers that act as bridge molecules between the silicon surface and DNA. The experiments until now performed suggest that the use of unsaturated organic acids protected or not appear to be good candidates for this chemistry.
Identification of ubiquitous mechanisms possessed by oxido-reductase activities involved in the control of reactive oxygen species (Prof. E. De Vendittis, Prof. M. Masullo, Dr. M. Ruocco) Superoxide dismutase (SOD), NADH/NADPH oxidase (NOX) and Thioredoxin reductase (TrxR) are key enzymes involved in the control of the physiological level of Reactive Oxygen Species (ROS). The research is aimed at the identification of possible common mechanisms of action through the study of the properties of these enzymes even in taxonomically distant organisms. A detailed characterization of the molecular, functional and structural properties of SOD from the hyperthermophilic archaeon Sulfolobus solfataricus has already been carried out. The research includes the Mn-SOD from rat mitochondria, and this study involves the production and characterization of wild-type and mutant forms of this enzyme. Furthermore, a SOD has been isolated and characterized from a psychrophilic micro-organism in order to study the adaptation in different growth conditions. Two distinct enzymes able to sustain a NOX activity have been isolated from S. solfataricus, but one has been reclassified as a TrxR. Moreover, the 3D-structure of wild-type and of a mutant form of TrxR have been solved in the presence of different cofactors. In the genome of S. Solfataricus three genes coding for two putative thioredoxin and another hypothetical thioredoxin reductase were found. The research will focus on the characterization of the thioredoxin system in S. solfataricus. The study will also include the characterization of NOX/TrxR activities from psychrophilic micro-organisms.
Biochemical and molecular characterization of the defects of the cholesterol biosynthesis (Prof. A. Dello Russo, Prof. G. Corso) Some congenital errors of the cholesterol biosynthesis have been found in humans. A classic example of these syndromes with congenital multiple anomalies and mental retardation (MCA/MR) is the Smith-Lemli-Opitz syndrome of (SLOS), due to the defect of 7-dehydrocholesterol reductase enzyme. Subsequently, other syndromes like the chondrodysplasia punctata X linked (CDPX2), the syndrome CHILD (sterol C4- demetyiase), the desmosterolosis (3-hydroxisterol-reductase) the dysplasia of Greenberg (sterol-C14 reductase) have been characterized. Recently our group has identified a new defect of the biosynthesis of the cholesterol in a patient affected from congenital multiple malformations, mental retardation and liver injury. In this patient we have found elevates levels in plasma and cells of lathosterol (lathosterolosis). We have demonstrated also that the reduced activity of the 3-hydroxysterol-5-desaturase was due to mutations of the corresponding gene (SC5D). In fact in the patient we have found 2 different mutations in trans on the two alleles. Currently we are working to search new defects that can be discovered by means of biochemical screening of patients affected by malformative diseases opportunely selected. Moreover, we are studying anomalous metabolites present in the lathosterolosis and/or in the patients affected from new defects of the cholesterol biosynthesis that could represent a start point in order to comprise the relationships between the malformative diseases, cholesterol biosynthesis and the embryonal development. In fact, in the patient affected from lathosterolosis we have found the presence of two anomalous metabolites. We are studying these compounds, sterols containing 28 and 29 carbon atoms, by mass spectrometry and spectroscopic nuclear magnetic resonance (N.M.R.). The results of this study could be useful in order to explain the pathogenetic role of the cholesterol deficit and/or of the accumulation of its precursors and/or the presence of anomalous metabolites. We have also transfected with both normal and mutated genes a K.O.S.C. strain (ERG3), and measured the enzymatic activity on the extracted microsomal fraction. We have found a significant correction of the defect of ERG3 when transfected with normal gene, we also suppose that the enzyme is nonspecific because the human enzyme reacts with endogenous ERG3 phytosterols. Finally, we are interested to search new defects by means of induction with inhibitors of enzymes of the cholesterol biosynthesis on cells in vitro.

Study of the role of immunophillins, and their inhibitor rapamycin, in cell survival of lymphoid neoplasias (Dr. M.F. Romano, Dr. R. Bisogni)) Our research is aimed at investigating the anti-leukemic effects of rapamycin possibly exerted independently of mTOR, through the inhibition immunophilins. First results of our research indicate that rapamycin, successfully targets two crucial signalling pathways involved in cell survival and chemoresistance of malignant lymphoblasts, i.e PI3k/Akt through mTOR and NF-kappaB through FKBP51, suggesting that this drug could improve the therapy of childhood ALL. We found that the immunosuppresant agent rapamycin was able to induce apoptosis of childhood acute lymphoblastic leukemia blasts saving normal hematopoietic cells. Although phosphatidylinositol triphosphate pathway has been widely found involved in tumorigenesis, our data showed that the block of this signaling pathway does not fully account for the efficacy of rapamycin against ALL, since the specific PI3k inhibitor wortmannin was less efficient in blast-killing compared to rapamycin. Therefore additional mechanisms appear to be involved in lymphoblast death that could be reasonably related to the FKBP-binding and -inhibiting property of the macrolide agent. Objectives Our project focuses on the anti-leukemic effects of rapamycin possibly exerted independently of mTOR, through the inhibition immunophilins. Methods The analysis of apoptosis is performed in flow cytometry, the study of gene expression through RT-PCR and western blot assay, the study of transcription factor activation is analysed by EMSA. The downmodulation of the immunophilins through siRNAs strategy will contribute to clarify their function. Results Although the block of phosphatidylinositol pathway appeared as being involved in the activation of death program of malignant lymphoblasts, nevertheless, rapamycin increased doxorubicin-induced apoptosis, even in non responder samples suggesting that further mechanisms were involved in the activity of rapamycin against ALL blasts. Anthracyclines activate NFkappaB and it is well known that disruption of this signalling pathway greatly increases the efficacy of apoptogenic stimuli. We demonstrated that FKBP51, the large immunophilin inhibited by rapamycin, was essential for drug-induced NF-kappaBactivation in human leukemia. Furthermore, we found that the macrolide was not able to increase doxorubicin-induced apoptosis in condition of NF-kappaB over-expression. In conclusion, our findings showed that rapamycin successfully targets two crucial signalling pathways involved in cell survival and chemoresistance of malignant lymphoblasts, i.e PI3k/Akt through mTOR and NF-kappaB through FKBP51, suggesting that this drug could improve the therapy of childhood ALL.

TECNOLOGIE IN POSSESSO DELL'U. O.

  • Heterologous protein expression
  • Methods of protein purification
  • 2D gel of proteins
  • Western blot analysis of protein expression
  • FT-IR and fluorescent spectroscopy
  • Recombinant DNA
  • Gene cloning and gene expression
  • RT-PCR
  • Cell culture and cell transformation
  • Cytofluorimetry

STRUMENTAZIONE

Denominazione
Struttura ove la strumentazione è allocata
Responsabile della strumentazione
Spettrofotometro UV-VIS a doppio raggio Cary 1E
Dipartimento di Biochimica e Biotecnologie Mediche
Antonio Dello Russo
Spettrofluorimetro Cary Eclipse
Dipartimento di Biochimica e Biotecnologie Mediche
Emmanuele De Vendittis
Spettrometro FT-IR con microscopia infrarosso Nicolet 5700
Dipartimento di Biochimica e Biotecnologie Mediche
Paolo Arcari
FPLC Pharmacia LCC-500 PLUS
Dipartimento di Biochimica e Biotecnologie Mediche
Rosaria Ruocco
Ultracentrifuga Beckman- Coulter Optima L-90K
Dipartimento di Biochimica e Biotecnologie Mediche
Maria Fiammetta Romano

PUBBLICAZIONI

1. L. De Stefano, L. Moretti, A. Lamberti, O. Longo, M. Rocchia, AM. Rossi, P. Arcari, I. Rendina. Optical Sensors for Vapors, Liquids, and Biological Molecules Based on Porous Silicon Technology . (2004) IEEE TRANS NANOTECHNOL 3, 49-54
2. L. Vitagliano, A. Ruggiero, M. Masullo, P. Cantiello, P. Arcari, A. Zagari. The Crystal Structure of Sulfolobus solfataricus Elongation Factor 1A in Complex with Magnesium and GDP. (2004) BIOCHEMISTRY-USA 43, 6630-6636
3. G. Raimo, B. Lombardo, M. Masullo, A. Lamberti, O. Longo, P. Arcari. Elongation Factor Ts from the Antarctic Eubacterium Pseudoalteromonas haloplanktis TAC 125: Biochemical Characterization and Cloning of the Encoding Gene. (2004) BIOCHEMISTRY-USA 43, 14759-14766
4. M.R. Ruocco, A. Ruggiero, L. Masullo, P. Arcari, M. Masullo. A 35 kDa NAD(P)H oxidase previously isolated from the archaeon Sulfolobus solfataricus is instead a thioredoxin reductase. (2004) BIOCHIMIE 86, 883-892
5. E. De Vendittis E, B. de Paola B, M.A. Gogliettino, B.S. Adinolfi, A. Fiengo, T. Duvold, V. Bocchini. Fusidic and helvolic acid inhibition of elongation factor 2 from the archaeon Sulfolobus solfataricus. (2002) BIOCHEMISTRY-USA 41, 14879-14884
6. MA. Gogliettino, F. Tanfani, A. Scire, T. Ursby, BS. Adinolfi, T. Cacciamani, E. De Vendittis. The Role of Tyr41 and His155 in the Functional Properties of Superoxide Dismutase from the Archaeon Sulfolobus solfataricus. (2004) BIOCHEMISTRY-USA 43, 2199-2208
7. MC. Turco, MF. Romano, A. Petrella, R. Bisogni, PF. Tassone, S. Venuta . NF-kB/Relmediated regulation of apoptosis in hematologic malignancies and normal hematopoietic progenitors. (2004) LEUKEMIA 18, 11-17
8. R. Avellino, S. Romano, R. Parasole, R. Bisogni, A. Lamberti, V. Poggi, S. Venuta, M. F. Romano . Rapamycin stimulates apoptosis of Childhood acute lymphoblastic leukemia cells . (2005) BLOOD 106, 1400-1406
9. G. Parenti, N. Brunetti-Pierri, G. Corso, M. Rossi, I. Annunziata, A. Battagliese, P. Ferrari, A. Ballabio, A. Dello Russo, G. Andria. Lathosterolemia, a novel defect of cholesterol biosynthesis in humans associated with multiple congenital malformation and mental retardation. (2002) J. INHER. METAB. DIS. 25:suppl-1(p:162).
10. M. Rossi, G. Federico, G. Corso, G. Parenti, A. Battagliese, A.R. Frascogna, R. Della Casa, A. Dello Russo, P. Strisciuglio, G. Saggese, G. Andria. Vitamin D status in patients affected by Smith-Lemli-Opitz syndrome. (2005) J. INHERIT. METAB. DIS. 28, 69-80.
LAMBERTI A., M. CARAGLIA, O. LONGO, M. MARRA, A. ABBRUZZESE, ARCARI P. . The translation elongation factor 1A in tumorigenesis, signal transduction and apoptosis: Review article. (2004) AMINO ACIDS.26, 443-448.
M. MASULLO, P. CANTIELLO, ARCARI P. Archaeal elongation factor 1 alpha from Sulfolobus solfataricus interacts with the eubacterial antibiotic GE2270A. (2004) EXTREMOPHILES. 8, pp. 499-505.
LUCA DE STEFANO, LUCIA ROTIROTI, ILARIA REA, LUIGI MORETTI, GIROLAMO DI FRANCIA, ETTORE MASSERA, ANNALISA LAMBERTI, ARCARI P., CARMEN SANGES AND IVO RENDINA. . Porous silicon-based optical biochips.(2006) JOURNAL OF OPTICS. A, PURE AND APPLIED OPTICS. 8, S540-S544.
RGGIERO I, RAIMO G, PALMA M, ARCARI P., MASULLO M. Molecular and functional properties of psychrophilic elongation factor G from the antarctic eubacterium Pseudoalteromonas haloplanktis TAC 125.(2007) EXTREMOPHILES 11, 699-709.
NARDONE G, RIPPA E, MARTIN G, ROCCO A, SICILIANO R.A, FIENGO A, CACACE G, MALORNI A, BUDILLON G, ARCARI P. Gastrokine 1 in patients with and without Helicobacter pylori infection.(2007) DIGESTIVE AND LIVER DISEASE. 39, 122-129.
LAMBERTI, A, LONGO O, MARRA M, TAGLIAFERRI P, BISMUTO E, FIENGO A, VISCOMI C, BUDILLON A, RAPP U.P. WANG E, VENUTA S, ABBRUZZESE A, ARCARI P., CARAGLIA M. C-Raf antagonizes apoptosis induced by INF-a in human lung cancer cells by phosphorylation and increase of the intracellular content of elongation factor 1A.(2007) CELL DEATH AND DIFFERENTIATION. 14, 952-962.
DE STEFANO L, ARCARI P., LAMBERTI A, SANGES C, ROTIROTI L, REA I, RENDINA I. DNA Optical Detection Based on Porous Silicon Technology: from Biosensors to Biochips. (2007) SENSORS (on line). 7, 214-221.
EMILIA RIPPA, GIOVANNI MARTIN, ALBA ROCCO, GIOVANNA LA MONICA, ANTONIO FIENGO, ROSA A, SICILIANO, GIUSI CACACE, ANTONIO MALORNI, GERARDO NARDONE, ARCARI P. Changes of protein expression in Helicobacter pylori-infected human gastric mucosa. (2007) CURRENT TOPICS IN BIOCHEMICAL RESEARCH. 8, 35-43.
ANNALISA LAMBERTI, MICHELE CARAGLIA, OLIMPIA LONGO, CARMEN SANGES, MONICA MARRA, ALBERTO ABBRUZZESE, ARCARI P. Functional role of translation Elongation Factor 1A in IFNa-induced apoptosis of human lung cancer cells. (2007) CURRENT TOPICS IN BIOCHEMICAL RESEARCH. 8, 51-60.

DOTTORATI DI RICERCA

Componente U.O. Dottorato di Ricerca Coordinatore Sede
Paolo Arcari
Biochimica e Biologia Cellulare e Molecolare

Università di Napoli Federrico II
Emmanuele De
Biochimica e Biologia Cellulare e Molecolare

Università di Napoli Federrico II
Antonio Dello Russo
Biochimica e Biologia Cellulare e Molecolare

Università di Napoli Federrico II

CONGRESSI C.I.B.

Congressi Partecipazione
CNB4

CNB5

CNB6

CNB7

CNB8

CNB9
CNB10