Staff: Prof. Malorni Antonio, Dr. Pocsfalvi Gabriella, Dr. Carbone Virginia, Dr. Siciliano Rosa, Dr. Perillo Bruno, Dr. Sorrentino Alida, Dr. De Giulio Beatrice, Faruolo Clementina, Ottombrino Antonio, Dr. De Caro Salvatore

No Staff: Dr. Cacace Giusi (Research fellow), Dr. Mazzeo Fiorella (Research fellow), Dr. Malorni Livia (PhD student), Dr. Vilasi Annalisa (PhD student), Saggese Paola (Student)

Following the decree of CNR n° 15422 of 13/12/1999 the International Mass Spectrometry Facilities Centre (CISESMA) was established at the site as a department of Institute of Food Science and Technology (ISA) of the C.N.R. in Avellino. All the instruments, including mass spectrometers and supporting techniques, were transferred from the laboratories of Naples to the new site at Avellino. The department gave also place to the direction of the Mass Spectrometry Network, an organization which brings together numerous southern Italian universities and CNR institutes and supported by the project "FERS Structural Funds1993-1999". The name of the department was changed by the Scientific Council of ISA to "Proteomic and Biomlecular Mass Spectrometry Centre (CeSMa-ProBio)" in September 2000. CeSMa-ProBio is equipped by eleven up to date mass spectrometers accompanied by various separation systems, some of them are unique in Italy and also rare in the world, such as the four-sector ZAB-T tandem mass spectrometer, the hybrid Autospec-oa-TOF tandem mass spectrometer and the complete "Proteomic System" of Micromass.

CeSMa-ProBio integrates the methodologies of modern mass spectrometry with the classical protein, carbohydrate and nucleotide chemistry and biochemistry. The activity is focused on the characterisation, optimisation and application of integrated approaches to resolve scientific problems of different chemical, biochemical, nutritional and environmental origin.

Research activities:

High-throughput proteomic studies based on the combined use of 2D gel electrophoresis and two-dimensional capillary chromatography and mass spectrometry. Development of a methodology for the rapid identification of bacteria by direct MALDI-TOF mass spectrometry analyses of intact micro-organisms. Set-up of an innovative and sensitive mass spectrometry based method for the analysis of selected bacterial protein toxins present in complex food matrixes Development of a new screening methodology using innovative peptide nucleic acid (PNA) in situ hybridization probes and biochip technology in combination with mass spectrometry detection. A novel approach based on the combined use of solid-phase affinity capture and mass spectrometry was applied to detect specific non-covalent interactions in model systems. Study of the modification of the biomacromolecules induced by genotoxic agents present in the environment. Structural characterization and post-translational modifications of proteins of relevant interest in food science , such as lattoferrin and glyadin. Isolation, purification and characterization of secondary metabolites present in complex matrixes. Determination of multi-element concentrations in food matrix using inductively coupled plasma high resolution sector field mass spectrometry ICP-MS. Microbiologic control of fermentative process in food production. Quality and safety food control. Microbic starters prodauction for milk products, bread and salted meats.

Fund obtained (2000-2003)

1. Fund:

Title of the research: Identification and functional analysis of molecular and genetic differences characteristic of hormone responsive breast cancer
Project co-ordinator: Prof. Alessandro Weisz
Ministry of Education, University and Research (PNR 2001-2003-FIRB art.8)

Research Unit ISA-AV:

Unit co-ordinator: Prof. Antonio Malorni

Proteomic studies of human breast cancer cell line ZR75.1 and evaluation of differences in protein expression profiles after estrogen stimulation

2. Fund:

Title of the research: Treatment of fresh highly perishable food stuff to guarantee quality, security and health (PROFSICURI)
Project co-ordinator: Prof. Raffaele Coppola
Ministry of University, Scientific Research and Technology (LEGGE 449/97-99)

Research Unit ISA-AV:

Unit co-ordinator: Prof. Antonio Malorni

Mass spectrometry methodologies for the rapid identification of bacteria

 3. Fund:

Title of the research: Hygienic security of typical Italian cheeses (SIFORTI)
Project co-ordinator: Dr. Germano Mucchetti
Ministry of University, Scientific Research and Technology (LEGGE 449/97-99)

Research Unit ISA-CNR:

Unit co-ordinator: Dr. Gabriella Pocsfalvi

Application of mass spectrometry to detect bacterial protein toxins

List of publication between 2000-2003

• 1. Minetti M., Pietraforte D., Carbone V., Salzano A. M., Scorza G. and Marino G. Scavenging of peroxynitrite by oxyemoglobin and identification of modified globin residues, Biochemistry 39 (22), 6689-6697, (2000).
• 2. Carbone V., Salzano A. M, Pagnini D., Papa G., Libertino R., Pagano L., De Rosa C. and Pucci P. The mutation associated with Hb Peterborough [b 111 (G13) Val(Phe] originated from Southern Italy, Hemoglobin 24 (3), 227-237, (2000).
• 3. Basile A., Ferranti P., Soleo L., Ambrosi L., Miraglia N., Malorni A., Sannolo N. Metodologie analitiche avanzate per ottimizzare la scelta di biomarker nella dosimetria molecolare degli addotti emoglobinici: il caso dell'1,3-butadiene, Folia Medica 71, 39-45, (2000).
• 4. Sannolo N., Miraglia N., Basile A., Ferranti P., Cuccovillo F., Acampora A., Malorni A. Lo studio dell'emoglobina modificata per la determinazione della dose biologicamente efficace: il caso del bromuro di metile Folia Medica, 71, 182-192, (2000).
• 5. Miraglia N., Pocsfalvi G., Ferranti P., Basile A., Sannolo N., Acampora A., Soleo L., Palmieri F., Malorni A. Caratterizzazione strutturale mediante tecniche di spettrometria di massa degli addotti emoglobina-epicloridrina a bassi livelli di concentrazione, Folia Medica, 71, 175-181, (2000).
• 6. Ferranti P., Mamone G. and Malorni A. Preparation and mass spectrometric analysis of S-nitrosohemoglobin, Methods Mol. Biol., 146, 147-165, (2000).
• 7. Denekamp C., Pócsfalvi G., Claeys M. Mechanism of Cross-Ring Cleavage Reactions in Dirhamnosyl Lipids: Effect of the Alkali Ion, Rapid. Commun. Mass Spectrom., 14, 794-799 (2000).
• 8. Lo Schiavo S., Pócsfalvi G., Serroni S., Cardiano G. P. and Piraino P. Self-Assembly of Square Molecular Boxes Containing Dirhodium(II,II) Units, Eur. J. Inorg. Chem., 6, 1371-1375 (2000).
• 9. Opsenica D., Pócsfalvi G., Jurani Z., Tinant B., Kyle D., Declercq J. P., Kyle D. E., Milhous W. K. and Solaja B. A. Cholic Acid Derivatives as 1,2,4,5-Tetraoxane Carriers: Structure and Antimalarial and Antiproliferative Activity, J. Med. Chem., 43, 3274-3282 (2000).
• 10. Pócsfalvi G., Ritieni A., Randazzo G., Dobó A. and Malorni A. Interaction of Fusarium Mycotoxins, Fusaproliferin and Fumonisin B1, with DNA Studied by Electrospray Ionization Mass Spectrometry, J. Agriculture and Food Chemistry , 48, 5794-5801 (2000).
• 11. Pócsfalvi G., Malorni A., Mancini I., Guella G. and Pietra F. Molecular Characterization of a Highly Heterogeneous Mixture of Glucosylceramides from a Deep-water Mediterranean Scleractinian Coral, Rapid Commun. Mass Spectrom., 14, 2247-225 (2000).
• 12. Cuyckens F., Ma Y. L., Pocsfalvi G. and Claeys M. Tandem mass spectral strategies for the structural characterization of flavonoid glycosides, Analusis, 28, 888-895 (2000).
• 13. Siciliano R.A., Rega B., Amoresano A. and Pucci P. Modern mass spectrometric methodologies in monitoring milk quality Anal. Chem. 72,408-415 (2000).
• 14. Amoresano A., Amedeo S., D'Andrea G., Siciliano R.A., Gagna C., Castagnaro M., Marino G. and Guarda, F. Structural characterization of oligosaccharide component of glycoproteins in normal and affected by endocardiosis pig mitral valves Eur. J. Biochem. 267,1299-1306 (2000).
• 15. Orru S., Amoresano A., Siciliano R.A., Napoleoni R., Finocchiaro O., Datola A., De Luca E., Sirna A. and Pucci P. Structural analysis of modified forms of recombinant IFN-beta produced under stress-simulating conditions. Biol. Chem. 381, 7-17 (2000).
• 16. Longobardo L., Melck D., Siciliano R.A., Santini A., Di Marzo V. and Cammarota G. Beta- casomorphins: substitution of phenylalanine with beta-homo phenylalanine increases the mu-type opioid receptor affinity Bioorg. Med. Chem. Lett. 10,1185-1188 (2000).
• 17. Miraglia N., Pocsfalvi G., Ferranti P., Basile A., Sannolo N., Acampora A., Soleo L., Palmieri F., Caira S., De Giulio B., Malorni A. Mass spectrometric identification of a candidate biomarker peptide from the in vitro interaction of epichlorohydrin with red blood cells, J. Mass Spectrom., 36, 47-57 (2001).
• 18. Basile A., Ferrante P., Pocsfalvi G., Mamone G., Miraglia N., Caira S., Ambrosi L., Soleo L., Sannolo N. and Malorni A. A Novel Approach for Detection and Measurement of Hemoglobin Adducts with 1,2,3,4 Diepoxibutane by LC/ES-MS and MALDI-MS/MS, Rapid Commun Mass Spectrom. 15, 527-540 (2001).
• 19. Carbone V., Salzano A. M., Pagano L., Buffardi S., De Rosa C. and Pucci P. Identification of Hb Villejuif [beta 123 (H1) Thr_Ile] in Southern Italy, Hemoglobin 25 (1), 67-78, (2001).
• 20. Montoro P., Carbone V., De Simone F., Pizza C., De Tommasi N. Studies on the constituent of Cyclanthera pedata fruits: isolation and structure elucidation of new flavonoid glycosides and their antioxidant activity, J. Agr. Food Chem. 49 (11), 5156-5160, (2001).
• 21. Di Maro A., Ferranti P., Mastronicola M., Polito L., Bolognesi A., Stirpe F., Malorni A. and Parente G. Reliable sequence determination of ribosome-inactivating proteins by combining electrospray mass spectrometry and Edman degradation, J. Mass Spectrom., 36: 38-46, (2001).
• 22. Metafora V., Franco P., Massa O., Morelli F., Stiuso P., Ferranti P., Mamone G., Malorni A., Stoppelli M. P. and Metafora S. Phosphorilation of seminal vescicle protein IV on Ser58 enhances its peroxidase-stimulating activity, Europ. J. Biochem., 268, 3858-3869, (2001).
• 23. Maurano F., Siciliano R.A., De Giulio B., Luongo D., Mazzeo M.F, Troncone R., Auricchio S. and Rossi M. Intranasal administration of one alpha gliadin can down-regulate the immune response to whole gliadin in mice. Scand. J. Immunol 53, 1-7 (2001).
• 24. Amoresano A., Orrù S., Siciliano R.A., De Luca E., Napoleoni R., Sirna A. and Pucci P. Assignment of the complete disulphide bridge pattern in the human recombinant follitropin beta-chain. Biol. Chem 382, 961-968 (2001).
• 25. Superti F., Siciliano R.A., Rega B., Giansanti F., Valenti P. and Antonini G. Involvement of bovine lactoferrin metal saturation, sialic acid and protein fragments in the inhibition of rotavirus infection. Biochim. Biophys. Acta 1528, 107-15 (2001).
• 26. Salzano A. M., Carbone V., Pagano L., Buffardi S., De Rosa C., Pucci P. Hb Vila Real [beta36(C2)Pro->His] in Italy: characterization of the amino acid substitution and the DNA mutation, Hemoglobin 26(1), 21-31, (2002).
• 27. Alaoui A., Charrouf Z., Pizza C., Carbone V., Malorni A., Piacente S. Triterpenoid Saponins from the shell of Argania spinosa seeds, J. Agr. Food Chem.; 50, 4600-4603, (2002).
• 28. Piacente S, Carbone V, Plaza A, Zampelli A, Pizza C. Investigation of the tuber constituents of maca (Lepidium meyenii Walp.), J. Agr. Food Chem.; 50, 5621-5, (2002).
• 29. Basile A., Ferranti P., Mamone G., Manco I., Pocsfalvi G., Malorni A., Acampora A., Sannolo N., Structural analysis of styrene oxide/haemoglobin adducts by mass spectrometry: identification of suitable biomarkers for human exposure evaluation, Rapid Commun, Mass Spectrom.;16(9), 871-8 (2002).
• 30. Solaja B. A., Terzic N., Pocsfalvi G., Gerena L., Tinant B., Opsenica D., Milhous W. K., Mixed steroidal 1,2,4,5-tetraoxanes: antimalarial and antimycobacterial activity, J. Med. Chem. 1;45(16):3331-6 (2002).
• 31. Miraglia N., Genovese G., Acampora A., Malorni L., Sannolo N., Ion trap mass spectrometry in the structural analysis of haemoglobin peptides modified by epichlorohydrin and diepoxybutane. Rapid communication in Mass Spectrometry 2002; 16 (9): 840-7.
• 32. Miraglia N., Basile A., Pieri M., Acampora A., Malorni L., De Giulio B., Sannolo N., A novel mass spectrometric technique for the evaluation of occupational exposure to methyl bromide. Journal of Mass Spectrometry 2002; 37 (11):1171-4.
• 33. Bandiera A., Tell G., Marsich E., Scaloni A., Pocsfalvi G., Akintunde Akindahunsi A., Cesaratto L., Manzini G. Cytosine-block telomeric type DNA-binding activity of hnRNP proteins from human cell lines, Arch. Biochem. Biophys. 15;409(2), 305-14 (2003).
• 34. Schlosser G., Takats Z., Vekey K., Pocsfalvi G., Malorni A., Windberg E., Kiss A., Hudecz F., Mass Spectrometric Analysis of Combinatorial Peptide Libraries Derived from the Tandem Repeat Unit of MUC2 Mucin, J. Peptide Sci., 9, 361-374 (2003).
• 35. Opsenica D., Angelovski G., Pocsfalvi G., Jurani_ Z., _ir_ak _., Kyle D., Milhous W. K., Solaja B. A.: Antimalarial and antiproliferative evaluation of Bis-Steroidal Tetraoxans, Bioorg. Med. Chem. 11, 2761-2768 (2003).