40 results on '"Pisano , Elisabetta"'
Search Results
2. La comunicazione nella/della ricerca, il caso CNR-IFC: uno studio pilota
- Author
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Pisano, Elisabetta, primary and Ferdeghini, Ezio-Maria, additional
- Published
- 2021
- Full Text
- View/download PDF
3. Detection and genotyping of human papillomavirus DNA in samples from healthy Sardinian patients: a preliminary study
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Montaldo, Caterina, Mastinu, Andrea, Quartuccio, Marta, Piras, Vincenzo, Denotti, Gloria, Pisano, Elisabetta, and Orrù, Germano
- Published
- 2007
4. Peptides of human gingival crevicular fluid determined by HPLC-ESI-MS
- Author
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Pisano, Elisabetta, Cabras, Tiziana, Montaldo, Caterina, Piras, Vincenzo, Inzitari, Rosanna, Olmi, Chiara, Castagnola, Massimo, and Messana, Irene
- Published
- 2005
5. N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M.
- Author
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Manconi, Barbara, Cabras, Tiziana, Sanna, Maria Teresa, Piras, V, Liori, B, Pisano, Elisabetta, Iavarone, Federica, Vincenzoni, Federica, Cordaro, Massimo, Faa, G, Castagnola, Massimo, Messana, Irene, Manconi , Barbara, Cabras , Tiziana, Sanna , Maria Teresa, Pisano , Elisabetta, Iavarone, Federica (ORCID:0000-0002-2074-5531), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene (ORCID:0000-0002-1436-6105), Manconi, Barbara, Cabras, Tiziana, Sanna, Maria Teresa, Piras, V, Liori, B, Pisano, Elisabetta, Iavarone, Federica, Vincenzoni, Federica, Cordaro, Massimo, Faa, G, Castagnola, Massimo, Messana, Irene, Manconi , Barbara, Cabras , Tiziana, Sanna , Maria Teresa, Pisano , Elisabetta, Iavarone, Federica (ORCID:0000-0002-2074-5531), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Castagnola, Massimo (ORCID:0000-0002-0959-7259), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213.
- Published
- 2016
6. Usefulness of real time PCR for the differentiation and quantification of 652 and JP2 Actinobacillus actinomycetemcomitans genotypes in dental plaque and saliva
- Author
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Piras Vincenzo, Baldoni Marco, Cotti Marina, Isola Daniela, Ciusa Maria, Marini Mario, Orrù Germano, Pisano Elisabetta, and Montaldo Caterina
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Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract Background The aim of our study is to describe a fast molecular method, able to distinguish and quantize the two different genotypes (652 and JP2) of an important periodontal pathogen: Actinobacillus actinomycetemcomitans. The two genotypes show differences in the expression of an important pathogenic factor: the leukotoxin (ltx). In order to evidence this, we performed a real time PCR procedure on the ltx operon, able to recognize Aa clinical isolates with different leukotoxic potentials. Methods The specificity of the method was confirmed in subgingival plaque and saliva specimens collected from eighty-one Italian (Sardinian) subjects with a mean age of 43.9, fifty five (68 %) of whom had various clinical forms of periodontal disease. Results This procedure showed a good sensitivity and a high linear dynamic range of quantization (107-102 cells/ml) for all genotypes and a good correlation factor (R2 = 0.97–0.98). Compared with traditional cultural methods, this real time PCR procedure is more sensitive; in fact in two subgingival plaque and two positive saliva specimens Aa was only detected with the molecular method. Conclusion A low number of Sardinian patients was found positive for Aa infections in the oral cavity, (just 10 positive periodontal cases out of 81 and two of these were also saliva positive). The highly leukotoxic JP2 strain was the most representative (60 % of the positive specimens); the samples from periodontal pockets and from saliva showed some ltx genotype for the same patient. Our experience suggests that this approach is suitable for a rapid and complete laboratory diagnosis for Aa infection.
- Published
- 2006
- Full Text
- View/download PDF
7. Chrono-proteomics of human saliva: variations of the salivary proteome during human development
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Messana, Irene, Cabras, Tiziana, Iavarone, Federica, Manconi, Barbara, Huang, Liling, Martelli, Claudia, Olianas, Alessandra, Sanna, Mt, Pisano, Elisabetta, Sanna, M, Arba, M, D'Alessandro, A, Desiderio, Claudia, Vitali, Alberto, Pirolli, Davide, Tirone, Chiara, Lio, Alessandra, Vento, Giovanni, Romagnoli, Costantino, Cordaro, Massimo, Manni, Armando, Gallenzi, Patrizia, Fiorita, Antonella, Scarano, Emanuele, Calo', Lea, Passali, Giulio Cesare, Picciotti, Pasqualina Maria, Paludetti, Gaetano, Fanos, V, Faa, G, Castagnola, Massimo, Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Iavarone, Federica (ORCID:0000-0002-2074-5531), Manconi , Barbara, Olianas , Alessandra, Pisano , Elisabetta, Pirolli, Davide (ORCID:0000-0003-2303-2577), Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Manni, Armando (ORCID:0000-0002-7784-1911), Gallenzi, Patrizia (ORCID:0000-0001-9805-4522), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Calo', Lea (ORCID:0000-0003-2671-336X), Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene, Cabras, Tiziana, Iavarone, Federica, Manconi, Barbara, Huang, Liling, Martelli, Claudia, Olianas, Alessandra, Sanna, Mt, Pisano, Elisabetta, Sanna, M, Arba, M, D'Alessandro, A, Desiderio, Claudia, Vitali, Alberto, Pirolli, Davide, Tirone, Chiara, Lio, Alessandra, Vento, Giovanni, Romagnoli, Costantino, Cordaro, Massimo, Manni, Armando, Gallenzi, Patrizia, Fiorita, Antonella, Scarano, Emanuele, Calo', Lea, Passali, Giulio Cesare, Picciotti, Pasqualina Maria, Paludetti, Gaetano, Fanos, V, Faa, G, Castagnola, Massimo, Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Iavarone, Federica (ORCID:0000-0002-2074-5531), Manconi , Barbara, Olianas , Alessandra, Pisano , Elisabetta, Pirolli, Davide (ORCID:0000-0003-2303-2577), Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Manni, Armando (ORCID:0000-0002-7784-1911), Gallenzi, Patrizia (ORCID:0000-0001-9805-4522), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Calo', Lea (ORCID:0000-0003-2671-336X), Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), and Castagnola, Massimo (ORCID:0000-0002-0959-7259)
- Abstract
An important contribution to the variability of any proteome is given by the time dimension that should be carefully considered to define physiological modifications. To this purpose, whole saliva proteome was investigated in a wide age range. Whole saliva was collected from 17 preterm newborns with a postconceptional age at birth of 178-217 days. In these subjects sample collection was performed serially starting immediately after birth and within about 1 year follow-up, gathering a total of 111 specimens. Furthermore, whole saliva was collected from 182 subjects aged between 0 and 17 years and from 23 adults aged between 27 and 57 years. The naturally occurring intact salivary proteome of the 316 samples was analyzed by low- and high-resolution HPLC-ESI-MS platforms. Proteins peculiar of the adults appeared in saliva with different time courses during human development. Acidic proline-rich proteins encoded by PRH2 locus and glycosylated basic proline-rich proteins encoded by PRB3 locus appeared following 180 days of postconceptional age, followed at 7 months (±2 weeks) by histatin 1, statherin, and P-B peptide. The other histatins and acidic proline-rich proteins encoded by PRH1 locus appeared in whole saliva of babies from 1 to 3 weeks after the normal term of delivery, S-type cystatins appeared at 1 year (±3 months), and basic proline-rich proteins appeared at 4 years (±1 year) of age. All of the proteinases involved in the maturation of salivary proteins were more active in preterm than in at-term newborns, on the basis of the truncated forms detected. The activity of the Fam20C kinase, involved in the phosphorylation of various proteins, started around 180 days of postconceptional age, slowly increased reaching values comparable to adults at about 2 years (±6 months) of age. Instead, MAPK14 involved in the phosphorylation of S100A9 was fully active since birth also in preterm newborns.
- Published
- 2015
8. N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M
- Author
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Manconi, Barbara, Cabras, Tiziana, Sanna, Maria Teresa, Piras, V, Liori, B, Pisano, Elisabetta, Iavarone, Federica, Vincenzoni, Federica, Cordaro, Massimo, Faa, G, Castagnola, Massimo, and Messana, Irene
- Subjects
Spectrometry, Mass, Electrospray Ionization ,Glycosylation ,Polysaccharides ,Protein ,Humans ,Settore BIO/10 - BIOCHIMICA ,Chromatography, High Pressure Liquid ,Salivary Proline-Rich Proteins - Abstract
In the present study, we show that the heterogeneous mixture of glycoforms of the basic salivary proline-rich protein 3M, encoded by PRB3-M locus, is a major component of the acidic soluble fraction of human whole saliva in the first years of life. Reversed-phase high-performance liquid chromatography with high-resolution electrospray ionization mass spectrometry analysis of the intact proteoforms before and after N-deglycosylation with Peptide-N-Glycosidase F and tandem mass spectrometry sequencing of peptides obtained after Endoproteinase GluC digestion allowed the structural characterization of the peptide backbone and identification of N- and O-glycosylation sites. The heterogeneous mixture of the proteoforms derives from the combination of 8 different neutral and sialylated glycans O-linked to Threonine 50, and 33 different glycans N-linked to Asparagine residues at positions 66, 87, 108, 129, 150, 171, 192, and 213.
- Published
- 2015
9. Top-down platform for deciphering the human salivary proteome
- Author
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Castagnola, Massimo, Cabras, Tiziana, Iavarone, Federica, Vincenzoni, Federica, Vitali, Andrea, Pisano, Elisabetta, Nemolato, S, Scarano, Emanuele, Fiorita, Antonella, Vento, Giovanni, Tirone, Chiara, Romagnoli, Costantino, Cordaro, Massimo, Paludetti, Gaetano, Faa, G, Messana, Irene, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Cabras , Tiziana, Iavarone, Federica (ORCID:0000-0002-2074-5531), Pisano , Elisabetta, Scarano, Emanuele (ORCID:0000-0003-2570-1121), Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Messana, Irene (ORCID:0000-0002-1436-6105), Castagnola, Massimo, Cabras, Tiziana, Iavarone, Federica, Vincenzoni, Federica, Vitali, Andrea, Pisano, Elisabetta, Nemolato, S, Scarano, Emanuele, Fiorita, Antonella, Vento, Giovanni, Tirone, Chiara, Romagnoli, Costantino, Cordaro, Massimo, Paludetti, Gaetano, Faa, G, Messana, Irene, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Cabras , Tiziana, Iavarone, Federica (ORCID:0000-0002-2074-5531), Pisano , Elisabetta, Scarano, Emanuele (ORCID:0000-0003-2570-1121), Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Cordaro, Massimo (ORCID:0000-0002-0797-5172), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
Proteomic platforms can be classified in bottom-up strategies, which analyze the sample after proteolytic digestion, and top-down strategies, which analyze the intact naturally occurring proteome. Bottom-up platforms are high-throughput because they can investigate a large number of proteins, regardless of their dimension. Nonetheless, information on post-translational modifications (PTMs) can be lost, especially those regarding naturally occurring cleavages and alternative splicing. Top-down platforms cannot cover vast proteomes, however, they can disclose subtle structural variations occurring during protein maturation and allow label-free relative quantifications in an unlimited number of samples. A repertoire of 256 masses belonging to naturally occurring proteins and peptides consistently detected by RP-HPLC-ESI-MS analysis of the acidic soluble fraction of human whole saliva is presented in this study. Of them, 233 have been identified, while 23 are still pending for the definitive characterization. The present review reports average and mono-isotopic masses of the peptides and proteins detected, RP-HPLC elution times, PTMs, origin and quali-quantitative variations observed in several physiological and pathological conditions. The information reported can be a reference for users of top-down RP-HPLC-ESI-MS proteomic platforms applied to the study of the human salivary proteome as well as of other human bodily fluids.
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- 2012
10. Potential applications of human saliva as diagnostic fluid
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Castagnola, Massimo, Picciotti, Pasqualina Maria, Messana, Irene, Fanali, Chiara, Fiorita, Antonella, Cabras, Tiziana, Calo', Lea, Pisano, Elisabetta, Passali, Giulio Cesare, Iavarone, Federica, Paludetti, Gaetano, Scarano, Emanuele, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Calo', Lea (ORCID:0000-0003-2671-336X), Pisano , Elisabetta, Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Iavarone, Federica (ORCID:0000-0002-2074-5531), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Castagnola, Massimo, Picciotti, Pasqualina Maria, Messana, Irene, Fanali, Chiara, Fiorita, Antonella, Cabras, Tiziana, Calo', Lea, Pisano, Elisabetta, Passali, Giulio Cesare, Iavarone, Federica, Paludetti, Gaetano, Scarano, Emanuele, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Calo', Lea (ORCID:0000-0003-2671-336X), Pisano , Elisabetta, Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Iavarone, Federica (ORCID:0000-0002-2074-5531), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), and Scarano, Emanuele (ORCID:0000-0003-2570-1121)
- Abstract
The use of human saliva as a diagnostic and prognostic fluid has until recently been somewhat disregarded. Although sample collection is non-invasive, physiological and genetic variations were largely responsible for its infrequent application in the past. Recently, several proteomic studies contributed to partial elucidation of the salivary proteome (more than 2400 protein components have been characterized), both in terms of composition, contributions to whole saliva and genetic/physiological variability. On this basis, is not too optimistic to believe that in the near future human saliva could become a relevant diagnostic fluid. In this review, the characterization by proteomic approaches of new salivary markers in oncology, head and neck carcinoma (oral cavity, oropharynx, larynx, and salivary glands), breast and gastric cancers, salivary gland function and disease, Sjögren syndrome, systemic sclerosis, dental and gingival pathology, systemic, psychiatric and neurological diseases, is described.
- Published
- 2011
11. Characterization of two isoforms of human SPRR3 from saliva of preterm human newborn and autoptic fetal oral mucosa, parotid and submandibular gland samples
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Manconi, Barbara, Cabras, Tiziana, Pisano, Elisabetta, Nemolato, S., Inzitari, Rosanna, Iavarone, Federica, Sanna, Maria Teresa, Tirone, Chiara, Vento, Giovanni, Romagnoli, Costantino, Faa, G., Castagnola, Massimo, Messana, Irene, Manconi , Barbara, Cabras , Tiziana, Pisano , Elisabetta, Iavarone, Federica (ORCID:0000-0002-2074-5531), Sanna , Maria Teresa, Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene (ORCID:0000-0002-1436-6105), Manconi, Barbara, Cabras, Tiziana, Pisano, Elisabetta, Nemolato, S., Inzitari, Rosanna, Iavarone, Federica, Sanna, Maria Teresa, Tirone, Chiara, Vento, Giovanni, Romagnoli, Costantino, Faa, G., Castagnola, Massimo, Messana, Irene, Manconi , Barbara, Cabras , Tiziana, Pisano , Elisabetta, Iavarone, Federica (ORCID:0000-0002-2074-5531), Sanna , Maria Teresa, Vento, Giovanni (ORCID:0000-0002-8132-5127), Romagnoli, Costantino (ORCID:0000-0003-1176-2943), Castagnola, Massimo (ORCID:0000-0002-0959-7259), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
RP-HPLC-ESI-MS profile of saliva samples from human preterm newborn showed a protein peak in the elution range 26.6-27.6min. Deconvolution of ESI-MS spectra revealed the presence of two proteins with average molecular mass (Mav) values of 17,239+/-3Da and 18,065+/-3Da in 9 samples, with Mav value of 17,239+/-3Da in 4 samples and Mav value of 18,065+/-3Da in 2 samples. MALDI-TOF-MS analysis of tryptic digest allowed identifying the proteins as two isoforms of small proline-rich protein 3 and cDNA amplification of RNA extracts from oral mucosa, parotid and submandibular gland samples, obtained at fetal autopsy, provided two nucleotide sequences in agreement with those reported in the literature. The two proteins differ for an octapeptide repeat (GCTKVPEP) and the substitution Leu-->Val, at position 148 and 140 of the mature form of the 18,065 and 17,239Da protein, respectively. During maturation the two proteins undergo two post-translational modifications, corresponding to N-terminal acetylation and removal of the initiator methionine. cDNA amplification did not allow to clarify if the proteins found in saliva originated from cellular shedding of the epithelium and/or secretion.
- Published
- 2010
12. Alteration of the salivary peptidome profile in children affected by type 1 diabetes.
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Cabras, Tiziana, Pisano, Elisabetta, Mastinu, Andrea, Denotti, Gloria, Pusceddu, Pietro Paolo, Inzitari, Rosanna, Fanali, Chiara, Nemolato, Sonia, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene (ORCID:0000-0002-1436-6105), Cabras, Tiziana, Pisano, Elisabetta, Mastinu, Andrea, Denotti, Gloria, Pusceddu, Pietro Paolo, Inzitari, Rosanna, Fanali, Chiara, Nemolato, Sonia, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
The acidic soluble fraction of whole saliva of type 1 diabetic children was analyzed by reversed phase (RP)(1)-HPLC-ESI-MS and compared with that of sex-and age-matched control subjects. Salivary acidic proline-rich phosphoproteins (aPRP), histatins, alpha-defensins, salivary cystatins, statherin, proline-rich peptide P-B (P-B), beta-thymosins, S100A8 and S100A9(star)(S100A9(star) corresponds to S100A9 vairant lacking the first four amino acids), as well some naturally occurring peptides derived from salivary acidic proline-rich phosphoproteins, histatins, statherin, and P-B peptide, were detected and quantified on the basis of the extracted ion current peak area. The level of phosphorylation of salivary acidic proline-rich phosphoproteins, histatin-1 (Hst-1), statherin and S100A9(star) and the percentage of truncated forms of salivary acidic proline-rich phosphoproteins was also determined in the two groups. The study revealed that statherin, proline-rich peptide P-B, P-C peptide, and histatins, were significantly less concentrated in saliva of diabetic subjects than in controls, while concentration of alpha-defensins 1, 2 and 4 and S100A9(star) was higher. The low concentration of P-C peptide was paralleled by high levels of some of its fragments. On the whole, the study highlighted the severe impairment of the repertoire of peptides involved in the safeguard of the oral cavity in children who have diabetes, as well as an higher concentration of the proinflammatory mediator S100A9(star) with respect to healthy children. Molecular & Cellular Proteomics 9:2099-2108, 2010.
- Published
- 2010
13. Age-Dependent Modifications of the Human Salivary Secretory Protein Complex
- Author
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Cabras, Tiziana, Pisano, Elisabetta, Boi, Roberto, Olianas, Alessandra, Manconi, Barbara, Inzitari, Rosanna, Fanali, Chiara, Giardina, Bruno, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Olianas , Alessandra, Manconi , Barbara, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene (ORCID:0000-0002-1436-6105), Cabras, Tiziana, Pisano, Elisabetta, Boi, Roberto, Olianas, Alessandra, Manconi, Barbara, Inzitari, Rosanna, Fanali, Chiara, Giardina, Bruno, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Olianas , Alessandra, Manconi , Barbara, Castagnola, Massimo (ORCID:0000-0002-0959-7259), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
Physiological variability of the naturally occurring, human salivary secretory peptidome was studied as a function of age. The qualitative and quantitative changes occurring in the secretion of proteins/peptides specific to the oral cavity (i.e., basic salivary proline-rich proteins, salivary acidic proline-rich phosphoproteins, statherin, proline-rich peptide P-B, salivary cystatins, and histatins) were investigated by high-performance liquid chromatography-electrospray ionization-mass spectrometry in 67 subjects aged between 3 and 44 years. Subjects were divided into five age groups: group A, 8 donors, 3-5 years; group B, 11 donors, 6-9 years; group C, 20 donors, 10-12 years; group D, 15 donors, 13-17 years; group E, 13 donors, 24-44 years. Basic salivary proline-rich proteins, almost undetectable in the 3-5 and 6-9 years groups, reached salivary levels comparable to that of adults (24-44 years) around puberty. Levels of peptide P-D, basic peptide P-F, peptide P-H, peptide P-J (a new basic salivary proline-rich protein characterized in this study), and basic proline-rich peptide IB-1 were significantly higher in the 10-12-year-old group than in the 3-5-year-old group, whereas the increase of proline-rich peptide II-2 was significant only after the age of 12 years. The concentration of salivary acidic proline-rich phosphoproteins, histatin-3 1/24, histatin-3 1/25, and monophosphorylated and diphosphorylated cystatin S showed a minimum in the 6-9-year-old group. Finally, the histatin-1 concentration was significantly higher in the youngest subjects (3-5 years) than in the other groups
- Published
- 2009
14. Alfa-Defensins levels in whole saliva of totally edentulous subjects
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Fanali, Chiara, Inzitari, Rosanna, Cabras, Tiziana, Pisano, Elisabetta, Castagnola, Massimo, Celletti, Renato, Manni, Armando, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Manni, Armando (ORCID:0000-0002-7784-1911), Messana, Irene (ORCID:0000-0002-1436-6105), Fanali, Chiara, Inzitari, Rosanna, Cabras, Tiziana, Pisano, Elisabetta, Castagnola, Massimo, Celletti, Renato, Manni, Armando, Messana, Irene, Cabras , Tiziana, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Manni, Armando (ORCID:0000-0002-7784-1911), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
Salivary levels of alpha-defensins 1-4 and histatins 1, 3 and 5 were determined in 11 totally edentulous patients, 11 younger healthy adults with normal gingival mucosa (Control group 1) and 8 subjects, age-matched with edentulous patients, having a minimum of 25 teeth (Control group II). Whole saliva was treated with trifluoroacetic acid and the acidic soluble fraction analyzed by High Performance Liquid Chromatography-Mass Spectrometry. The area of the extracted ion current peaks was used for peptide quantification. Levels of alpha-defensinsl-4, but not of histatins, were significantly lower in totally edentulous patients with respect to both Control group I and Control group II. The two control groups did not show significant differences. The reduced level of oral alpha-defensins, which are mainly of crevicular origin, is most likely due to the absence of the gingival sulcus in the edentulous subjects. The near absence of alpha-defensins might be in part responsible for the higher vulnerability of the oral cavity to oral pathogen infections observed in totally edentulous patients.
- Published
- 2008
15. Trafficking and post-secretory events responsible for the formation of secreted human salivary peptides. A proteomic approach
- Author
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Messana, Irene, Cabras, Tiziana, Pisano, Elisabetta, Sanna, Maria Teresa, Olianas, Alessandra, Manconi, Barbara, Pellegrini, Magi, Paludetti, Gaetano, Scarano, Emanuele, Fiorita, Antonella, Agostino, Stefania, Contucci, Alessia Maria, Calo', Lea, Picciotti, Pasqualina Maria, Manni, Armando, Bennick, Ander, Vitali, Alberto, Fanali, Chiara, Inzitari, Rosanna, Castagnola, Massimo, Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Pisano , Elisabetta, Sanna , Maria Teresa, Olianas , Alessandra, Manconi , Barbara, Pellegrini , Magi, Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Calo', Lea (ORCID:0000-0003-2671-336X), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Manni, Armando (ORCID:0000-0002-7784-1911), Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene, Cabras, Tiziana, Pisano, Elisabetta, Sanna, Maria Teresa, Olianas, Alessandra, Manconi, Barbara, Pellegrini, Magi, Paludetti, Gaetano, Scarano, Emanuele, Fiorita, Antonella, Agostino, Stefania, Contucci, Alessia Maria, Calo', Lea, Picciotti, Pasqualina Maria, Manni, Armando, Bennick, Ander, Vitali, Alberto, Fanali, Chiara, Inzitari, Rosanna, Castagnola, Massimo, Messana, Irene (ORCID:0000-0002-1436-6105), Cabras , Tiziana, Pisano , Elisabetta, Sanna , Maria Teresa, Olianas , Alessandra, Manconi , Barbara, Pellegrini , Magi, Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Calo', Lea (ORCID:0000-0003-2671-336X), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Manni, Armando (ORCID:0000-0002-7784-1911), and Castagnola, Massimo (ORCID:0000-0002-0959-7259)
- Abstract
To elucidate the localization of post-translational modifications of different classes of human salivary proteins and peptides (acidic and basic proline-rich proteins (PRPs), Histatins, Statherin, P-B peptide, and "S type" Cystatins) a comparative reversed phase HPLC-ESI-MS analysis on intact proteins of enriched granule preparations from parotid and submandibular glands as well as parotid, submandibular/sublingual (Sm/SI), and whole saliva was performed. The main results of this study indicate the following. (1) Phosphorylation of all salivary peptides, sulfation of Histatin 1, proteolytic cleavages of acidic and precursor basic PRPs occur before granule storage. (ii) In agreement with previous studies, basic PRPs are secreted by the parotid gland only, whereas all isoforms of acidic PRPs (aPRPs) are secreted by both parotid and Sm/SI glands. (iii) Phosphorylation levels of aPRPs, Histatin 1, and Statherin are higher in the parotid gland, whereas the extent of cleavage of aPRP is higher in Sm/SI glands. (iv) O-Sulfation of tyrosines of Histatin 1 is a posttranslational modification specific for the submandibular gland. (v) The concentration of Histatin 3, Histatin 5, and Histatin 6, but not Histatin 1, is higher in parotid saliva. (vi) Histatin 3 is submitted to the first proteolytic cleavage (generating Histatins 6 and 5) during granule maturation, and it occurs to the same relative extent in both glands. (vii) The proteolytic cleavages of Histatin 5 and 6, generating a cascade of Histatin 3 fragments, take place after granule secretion and are more extensive in parotid secretion. (viii) Basic PRPs are cleaved in the oral cavity by unknown peptidases, generating various small proline-rich peptides. (ix) C-terminal removal from Statherin is more extensive in parotid saliva. (x) P-B peptide is secreted by both glands, and its relative quantity is higher in submandibular/sublingual secretion. (xi) In agreement with previous studies, S type Cystatins are mainly the pr
- Published
- 2008
16. HPLC-MS characterization of cyclo-statherin Q37, a specific cyclization product of human salivary statherin generated by transglutaminase 2
- Author
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Cabras, Tiziana, Inzitari, Rosanna, Fanali, Chiara, Scarano, Emanuele, Patamia, Maria, Sanna, Maria Teresa, Pisano, Elisabetta, Giardina, Bruno, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Scarano, Emanuele (ORCID:0000-0003-2570-1121), Sanna , Maria Teresa, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), Messana, Irene (ORCID:0000-0002-1436-6105), Cabras, Tiziana, Inzitari, Rosanna, Fanali, Chiara, Scarano, Emanuele, Patamia, Maria, Sanna, Maria Teresa, Pisano, Elisabetta, Giardina, Bruno, Castagnola, Massimo, Messana, Irene, Cabras , Tiziana, Scarano, Emanuele (ORCID:0000-0003-2570-1121), Sanna , Maria Teresa, Pisano , Elisabetta, Castagnola, Massimo (ORCID:0000-0002-0959-7259), and Messana, Irene (ORCID:0000-0002-1436-6105)
- Abstract
In the present study the analytical potential of HPLC-MS/MS was utilized for the structural characterization of a post-translational modification of statherin. Human salivary statherin (M-av 5380.0 +/- 0.3 Da) is transformed by the action of transglutaminase 2 into a cyclic derivative with an average molecular mass of 5363.0 +/- 0.3 Da. The intra-molecular bridge is generated by the loss of an ammonia molecule between the unique lone-pair donating nucleophile Lys-6 and one acceptor among the seven glutamine residues of statherin. Digestion of the cyclic derivative with chymotrypsin, proteinase K, and carboxypeptidase Y, monitored by HPLC - electrospray ionization-ion trap-mass spectrometric analysis, demonstrated that cyclization involved almost specifically Gln-37 (> 95%), with the percentage of Gln-39 implicated in the cross-linking being less than 5%. The main derivative was named cyclo-statherin Q-37. Guinea pig transglutaminase 2 showed high affinity for statherin in vitro (K-m = 0.65 +/- 0.06 mu M). Cyclo-statherin was detected in vivo by HPLC-electros pray ionization ion trap-mass spectrometry analysis of whole human saliva and it accounted for about 1% of total statherin. Detection of cyclo-statherin in whole saliva is suggestive of a putative role of this molecule in the formation of the "oral protein pellicle".
- Published
- 2006
17. N- and O-linked glycosylation site profiling of the human basic salivary proline-rich protein 3M
- Author
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Manconi, Barbara, primary, Cabras, Tiziana, additional, Sanna, Monica, additional, Piras, Valentina, additional, Liori, Barbara, additional, Pisano, Elisabetta, additional, Iavarone, Federica, additional, Vincenzoni, Federica, additional, Cordaro, Massimo, additional, Faa, Gavino, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2016
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18. Chrono-Proteomics of Human Saliva: Variations of the Salivary Proteome during Human Development
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Messana, Irene, primary, Cabras, Tiziana, additional, Iavarone, Federica, additional, Manconi, Barbara, additional, Huang, Liling, additional, Martelli, Claudia, additional, Olianas, Alessandra, additional, Sanna, Maria Teresa, additional, Pisano, Elisabetta, additional, Sanna, Monica, additional, Arba, Morena, additional, D’Alessandro, Alfredo, additional, Desiderio, Claudia, additional, Vitali, Alberto, additional, Pirolli, Davide, additional, Tirone, Chiara, additional, Lio, Alessandra, additional, Vento, Giovanni, additional, Romagnoli, Costantino, additional, Cordaro, Massimo, additional, Manni, Armando, additional, Gallenzi, Patrizia, additional, Fiorita, Antonella, additional, Scarano, Emanuele, additional, Calò, Lea, additional, Passali, Giulio Cesare, additional, Picciotti, Pasqualina Maria, additional, Paludetti, Gaetano, additional, Fanos, Vassilios, additional, Faa, Gavino, additional, and Castagnola, Massimo, additional
- Published
- 2015
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19. Distribution of human papillomavirus genotypes in sardinian patients with oral squamous cell carcinoma
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Montaldo, Caterina, Mastinu, Andrea, Zorco, Stefania, Santini, Noemi, Pisano, Elisabetta, Piras, Vincenzo, Denotti, Gloria, Peluffo, Carla, Erriu, Matteo, Garau, Valentino, and Orrù, Germano
- Subjects
oral squamous cell carcinoma ,stomatognathic diseases ,HPV ,seminested PCR ,virus diseases ,Sardinian patients ,Article ,female genital diseases and pregnancy complications - Abstract
Human papillomaviruses (HPVs) seem to play an important role in the pathogenesis of gynecological carcinomas and in head and neck carcinomas. The aim of this study was to detect and genotype HPVs in fresh oral squamous cell carcinoma (OSCC) from a Sardinian population, and to determine whether HPV presence was significantly associated with the development of OSCC. The oral mucosa tissues were obtained from 120 samples (68 OSCC and 52 control samples) taken from a Sardinian population seen at the Dental Clinic of the Department of Surgery and Odontostomatological Sciences, University of Cagliari (Italy) and the “ Ospedale SS Trinità”, Cagliari (A.S.L. 8) between 2007 and 2008. PCR was used for the detection of HPV DNA and the genotype was determined by DNA sequencing. The frequency of HPV infection was evaluated in relation to age, sex, smoking and alcohol use. Statistical analysis was performed using the SPSS 11.5 software. The results showed the presence of HPV-DNA in 60.3% of OSCC with HPV-16 (51.2%) being the most frequent genotype. In these Sardinian OSCC patients, HPV-DNA was detected more in males (65.8%) than in females (34.1%) while controls show a 0% of HPV presence. HPV positive was highly associated with OSCC among subjects with a history of heavy tobacco and alcohol use and among those with no such history. A greater frequency of high risk HPV presence was observed in patients with OSCC compared to health control patients. In addition these results suggested that oral HPV presence could be associated in OSCC subjects. Our results need more analyses to detect the HPV-DNA integration into tumoral cells.
- Published
- 2010
20. Potential applications of human saliva as diagnostic fluid
- Author
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Castagnola, Massimo (ORCID:0000-0002-0959-7259), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Messana, Irene (ORCID:0000-0002-1436-6105), Fanali, Chiara, Fiorita, Antonella, Cabras, Tiziana, Calo', Lea (ORCID:0000-0003-2671-336X), Pisano, Elisabetta, Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Iavarone, Federica (ORCID:0000-0002-2074-5531), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), Scarano, Emanuele (ORCID:0000-0003-2570-1121), Castagnola, Massimo (ORCID:0000-0002-0959-7259), Picciotti, Pasqualina Maria (ORCID:0000-0002-1502-6508), Messana, Irene (ORCID:0000-0002-1436-6105), Fanali, Chiara, Fiorita, Antonella, Cabras, Tiziana, Calo', Lea (ORCID:0000-0003-2671-336X), Pisano, Elisabetta, Passali, Giulio Cesare (ORCID:0000-0002-8176-0962), Iavarone, Federica (ORCID:0000-0002-2074-5531), Paludetti, Gaetano (ORCID:0000-0003-2480-1243), and Scarano, Emanuele (ORCID:0000-0003-2570-1121)
- Abstract
The use of human saliva as a diagnostic and prognostic fluid has until recently been somewhat disregarded. Although sample collection is non-invasive, physiological and genetic variations were largely responsible for its infrequent application in the past. Recently, several proteomic studies contributed to partial elucidation of the salivary proteome (more than 2400 protein components have been characterized), both in terms of composition, contributions to whole saliva and genetic/physiological variability. On this basis, is not too optimistic to believe that in the near future human saliva could become a relevant diagnostic fluid. In this review, the characterization by proteomic approaches of new salivary markers in oncology, head and neck carcinoma (oral cavity, oropharynx, larynx, and salivary glands), breast and gastric cancers, salivary gland function and disease, Sjögren syndrome, systemic sclerosis, dental and gingival pathology, systemic, psychiatric and neurological diseases, is described.
- Published
- 2011
21. Significant Modifications of the Salivary Proteome Potentially Associated with Complications of Down Syndrome Revealed by Top-down Proteomics
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Cabras, Tiziana, primary, Pisano, Elisabetta, additional, Montaldo, Caterina, additional, Giuca, Maria Rita, additional, Iavarone, Federica, additional, Zampino, Giuseppe, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2013
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22. Top-down HPLC–ESI–MS detection of S-Glutathionylated and S-Cysteinylated Derivatives of Cystatin B and Its 1–53 and 54–98 Fragments in Whole Saliva of Human Preterm Newborns
- Author
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Iavarone, Federica, primary, Cabras, Tiziana, additional, Pisano, Elisabetta, additional, Sanna, Maria Teresa, additional, Nemolato, Sonia, additional, Vento, Giovanni, additional, Tirone, Chiara, additional, Romagnoli, Costantino, additional, Cordaro, Massimo, additional, Fanos, Vassilios, additional, Faa, Gavino, additional, Messana, Irene, additional, and Castagnola, Massimo, additional
- Published
- 2013
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23. Top-down platform for deciphering the human salivary proteome
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Castagnola, Massimo, primary, Cabras, Tiziana, additional, Iavarone, Federica, additional, Vincenzoni, Federica, additional, Vitali, Alberto, additional, Pisano, Elisabetta, additional, Nemolato, Sonia, additional, Scarano, Emanuele, additional, Fiorita, Antonella, additional, Vento, Giovanni, additional, Tirone, Chiara, additional, Romagnoli, Costantino, additional, Cordaro, Massimo, additional, Paludetti, Gaetano, additional, Faa, Gavino, additional, and Messana, Irene, additional
- Published
- 2012
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- View/download PDF
24. HPLC-ESI-MS and MS/MS structural characterization of multifucosylated N-glycoforms of the basic proline-rich protein IB-8a CON1+ in human saliva
- Author
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Cabras, Tiziana, primary, Boi, Roberto, additional, Pisano, Elisabetta, additional, Iavarone, Federica, additional, Fanali, Chiara, additional, Nemolato, Sonia, additional, Faa, Gavino, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2012
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25. The Surprising Composition of the Salivary Proteome of Preterm Human Newborn
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Castagnola, Massimo, primary, Inzitari, Rosanna, additional, Fanali, Chiara, additional, Iavarone, Federica, additional, Vitali, Alberto, additional, Desiderio, Claudia, additional, Vento, Giovanni, additional, Tirone, Chiara, additional, Romagnoli, Costantino, additional, Cabras, Tiziana, additional, Manconi, Barbara, additional, Teresa Sanna, Maria, additional, Boi, Roberto, additional, Pisano, Elisabetta, additional, Olianas, Alessandra, additional, Pellegrini, Mariagiuseppina, additional, Nemolato, Sonia, additional, Wilhelm Heizmann, Claus, additional, Faa, Gavino, additional, and Messana, Irene, additional
- Published
- 2011
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26. Alterations of the Salivary Secretory Peptidome Profile in Children Affected by Type 1 Diabetes
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Cabras, Tiziana, primary, Pisano, Elisabetta, additional, Mastinu, Andrea, additional, Denotti, Gloria, additional, Pusceddu, Pietro Paolo, additional, Inzitari, Rosanna, additional, Fanali, Chiara, additional, Nemolato, Sonia, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2010
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27. Distribution of Human Papillomavirus Genotypes in Sardinian Patients with Oral Squamous Cell Carcinoma~!2010-03-01~!2010-05-10~!2010-07-13~!
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Montaldo, Caterina, primary, Mastinu, Andrea, additional, Zorco, Stefania, additional, Santini, Noemi, additional, Pisano, Elisabetta, additional, Piras, Vincenzo, additional, Denotti, Gloria, additional, Peluffo, Carla, additional, Erriu, Matteo, additional, Garau, Valentino, additional, and Orru, Germano, additional
- Published
- 2010
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- View/download PDF
28. Characterization of two isoforms of human SPRR3 from saliva of preterm human newborn and autoptic fetal oral mucosa, parotid and submandibular gland samples
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Manconi, Barbara, primary, Cabras, Tiziana, additional, Pisano, Elisabetta, additional, Nemolato, Sonia, additional, Inzitari, Rosanna, additional, Iavarone, Federica, additional, Fanali, Chiara, additional, Sanna, Maria Teresa, additional, Tirone, Chiara, additional, Vento, Giovanni, additional, Romagnoli, Costantino, additional, Faa, Gavino, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2010
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- View/download PDF
29. Age-Dependent Modifications of the Human Salivary Secretory Protein Complex
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Cabras, Tiziana, primary, Pisano, Elisabetta, additional, Boi, Roberto, additional, Olianas, Alessandra, additional, Manconi, Barbara, additional, Inzitari, Rosanna, additional, Fanali, Chiara, additional, Giardina, Bruno, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2009
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30. HPLC-ESI-MS analysis of oral human fluids reveals that gingival crevicular fluid is the main source of oral thymosins β4and β10
- Author
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Inzitari, Rosanna, primary, Cabras, Tiziana, additional, Pisano, Elisabetta, additional, Fanali, Chiara, additional, Manconi, Barbara, additional, Scarano, Emanuele, additional, Fiorita, Antonella, additional, Paludetti, Gaetano, additional, Manni, Armando, additional, Nemolato, Sonia, additional, Faa, Gavino, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2008
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31. Trafficking and Postsecretory Events Responsible for the Formation of Secreted Human Salivary Peptides
- Author
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Messana, Irene, primary, Cabras, Tiziana, additional, Pisano, Elisabetta, additional, Sanna, Maria Teresa, additional, Olianas, Alessandra, additional, Manconi, Barbara, additional, Pellegrini, Mariagiuseppina, additional, Paludetti, Gaetano, additional, Scarano, Emanuele, additional, Fiorita, Antonella, additional, Agostino, Stefania, additional, Contucci, Alessia M., additional, Calò, Lea, additional, Picciotti, Pasqualina M., additional, Manni, Armando, additional, Bennick, Anders, additional, Vitali, Alberto, additional, Fanali, Chiara, additional, Inzitari, Rosanna, additional, and Castagnola, Massimo, additional
- Published
- 2008
- Full Text
- View/download PDF
32. HPLC–MS characterization of cyclo-statherin Q-37, a specific cyclization product of human salivary statherin generated by transglutaminase 2
- Author
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Cabras, Tiziana, primary, Inzitari, Rosanna, additional, Fanali, Chiara, additional, Scarano, Emanuele, additional, Patamia, Maria, additional, Sanna, Maria T., additional, Pisano, Elisabetta, additional, Giardina, Bruno, additional, Castagnola, Massimo, additional, and Messana, Irene, additional
- Published
- 2006
- Full Text
- View/download PDF
33. Usefulness of real time PCR for the differentiation and quantification of 652 and JP2 Actinobacillus actinomycetemcomitans genotypes in dental plaque and saliva
- Author
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Orrù, Germano, primary, Marini, Mario Francesco, additional, Ciusa, Maria Laura, additional, Isola, Daniela, additional, Cotti, Marina, additional, Baldoni, Marco, additional, Piras, Vincenzo, additional, Pisano, Elisabetta, additional, and Montaldo, Caterina, additional
- Published
- 2006
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34. HPLC- ESI- MS and MS/ MS structural characterization of multifucosylated N-glycoforms of the basic proline-rich protein IB-8a CON1+ in human saliva.
- Author
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Cabras, Tiziana, Boi, Roberto, Pisano, Elisabetta, Iavarone, Federica, Fanali, Chiara, Nemolato, Sonia, Faa, Gavino, Castagnola, Massimo, and Messana, Irene
- Abstract
This study describes the characterization of the glycan moieties and the peptide backbone of six glycoforms of IB-8a CON1
+ , a basic proline-rich protein present in human saliva. MS analyses on the intact glycoproteins before and after N-deglycosylation with PNGase F and high-resolution MS/ MS sequencing by LTQ Orbitrap XL of peptides and glycopeptides from tryptic digests allowed the structural characterization of the glycan moieties and the polypeptide backbone, as well as to establish the glycosylation site at the asparagine residue at 98th position. Five of the glycoforms carry a biantennary N-linked glycan fucosylated in the innermost N-acetylglucosamine of the core and showing from zero to four additional fucoses in the antennal region. The sixth glycoform carries a monoantennary monofucosylated oligosaccharide. The glycoform cluster was detected on 28 of 71 adult saliva specimens. Level of fucosylation showed interindividual variability with the major relative abundance for the trifucosylated glycoform. Nonglycosylated IB-8a CON1+ and the variant IB-8a CON1− , lacking of the glycosylation site, have been also detected in human saliva. [ABSTRACT FROM AUTHOR]- Published
- 2012
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- View/download PDF
35. HPLC‐ESI‐MSand MS/MSstructural characterization of multifucosylated N‐glycoforms of the basic proline‐rich protein IB‐8a CON1+in human saliva
- Author
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Cabras, Tiziana, Boi, Roberto, Pisano, Elisabetta, Iavarone, Federica, Fanali, Chiara, Nemolato, Sonia, Faa, Gavino, Castagnola, Massimo, and Messana, Irene
- Abstract
This study describes the characterization of the glycan moieties and the peptide backbone of six glycoforms of IB‐8a CON1+, a basic proline‐rich protein present in human saliva. MSanalyses on the intact glycoproteins before and after N‐deglycosylation with PNGase Fand high‐resolution MS/MSsequencing by LTQOrbitrap XLof peptides and glycopeptides from tryptic digests allowed the structural characterization of the glycan moieties and the polypeptide backbone, as well as to establish the glycosylation site at the asparagine residue at 98th position. Five of the glycoforms carry a biantennary N‐linked glycan fucosylated in the innermost N‐acetylglucosamine of the core and showing from zero to four additional fucoses in the antennal region. The sixth glycoform carries a monoantennary monofucosylated oligosaccharide. The glycoform cluster was detected on 28 of 71 adult saliva specimens. Level of fucosylation showed interindividual variability with the major relative abundance for the trifucosylated glycoform. Nonglycosylated IB‐8a CON1+and the variant IB‐8a CON1−, lacking of the glycosylation site, have been also detected in human saliva.
- Published
- 2012
- Full Text
- View/download PDF
36. HPLC‐ESI‐MS analysis of oral human fluids reveals that gingival crevicular fluid is the main source of oral thymosins β4and β10
- Author
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Inzitari, Rosanna, Cabras, Tiziana, Pisano, Elisabetta, Fanali, Chiara, Manconi, Barbara, Scarano, Emanuele, Fiorita, Antonella, Paludetti, Gaetano, Manni, Armando, Nemolato, Sonia, Faa, Gavino, Castagnola, Massimo, and Messana, Irene
- Abstract
Thymosin β4(Tβ4), its sulfoxide, and thymosin β10 (Tβ10) were detected in human saliva and identified by different strategies based on RP HPLC coupled to electrospray multidimensional IT MS. Tβ4 was almost always detected in whole saliva, its sulfoxide sporadically, Tβ10rarely. Tβ4was undetectable in parotid saliva and less concentrated in submandibular/sublingual saliva than in whole saliva. Analysis of gingival crevicular fluid revealed high relative amounts of Tβ4, Tβ4sulfoxide, and Tβ10in all the samples. Tβ4mean concentration was 200 times higher in crevicular fluid (20 μmol/L, N= 9) than in whole saliva (0.1 μmol/L, N= 9). Crevicular fluid concentration of Tβ4 (ca. 5% represented by its sulfoxide) and β10 significantly correlated (r= 0.856; N= 9), and their ratio was about 5. A significant correlation was also observed between Tβ4 concentrations in whole saliva and gingival crevicular fluid (r= 0.738;N= 9). Immunohistochemical analysis of the major salivary glands showed that immunoreactivity for Tβ4is restricted to ductal cells, with minor degree of focal positivity in some acinar cells. On the whole, results indicate that gingival sulcus is a main, although not the sole, source for oral Tβ4 and Tβ10.
- Published
- 2009
- Full Text
- View/download PDF
37. Trafficking and post-secretory events responsible for the formation of secreted human salivary peptides. A proteomic approach
- Author
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Irene Messana, Cabras, Tiziana, Pisano, Elisabetta, Sanna, Maria Teresa, Olianas, Alessandra, Manconi, Barbara, Pellegrini, Magi, Gaetano Paludetti, Scarano, Emanuele, Fiorita, Antonella, Agostino, Stefania, Contucci, Alessia Maria, Lea CALO', Picciotti, Pasqualina Maria, Manni, Armando, Bennick, Anders, Vitali, Alberto, Fanali, Chiara, Inzitari, Rosanna, and Massimo Castagnola
- Subjects
Proteomics ,Spectrometry, Mass, Electrospray Ionization ,SALIVARY ,Sulfates ,PROTEOMIC ,Molecular Sequence Data ,Protein Transport ,Humans ,Parotid Gland ,Amino Acid Sequence ,Phosphorylation ,Salivary Proteins and Peptides ,Peptides ,Saliva ,Settore BIO/10 - BIOCHIMICA ,Protein Processing, Post-Translational ,Alleles ,Chromatography, High Pressure Liquid - Abstract
To elucidate the localization of post-translational modifications of different classes of human salivary proteins and peptides (acidic and basic proline-rich proteins (PRPs), Histatins, Statherin, P-B peptide, and "S type" Cystatins) a comparative reversed phase HPLC-ESI-MS analysis on intact proteins of enriched granule preparations from parotid and submandibular glands as well as parotid, submandibular/sublingual (Sm/Sl), and whole saliva was performed. The main results of this study indicate the following. (i) Phosphorylation of all salivary peptides, sulfation of Histatin 1, proteolytic cleavages of acidic and precursor basic PRPs occur before granule storage. (ii) In agreement with previous studies, basic PRPs are secreted by the parotid gland only, whereas all isoforms of acidic PRPs (aPRPs) are secreted by both parotid and Sm/Sl glands. (iii) Phosphorylation levels of aPRPs, Histatin 1, and Statherin are higher in the parotid gland, whereas the extent of cleavage of aPRP is higher in Sm/Sl glands. (iv) O-Sulfation of tyrosines of Histatin 1 is a post-translational modification specific for the submandibular gland. (v) The concentration of Histatin 3, Histatin 5, and Histatin 6, but not Histatin 1, is higher in parotid saliva. (vi) Histatin 3 is submitted to the first proteolytic cleavage (generating Histatins 6 and 5) during granule maturation, and it occurs to the same relative extent in both glands. (vii) The proteolytic cleavages of Histatin 5 and 6, generating a cascade of Histatin 3 fragments, take place after granule secretion and are more extensive in parotid secretion. (viii) Basic PRPs are cleaved in the oral cavity by unknown peptidases, generating various small proline-rich peptides. (ix) C-terminal removal from Statherin is more extensive in parotid saliva. (x) P-B peptide is secreted by both glands, and its relative quantity is higher in submandibular/sublingual secretion. (xi) In agreement with previous studies, S type Cystatins are mainly the product of Sm/Sl glands.
38. Monitoraggio dell'attività motoria mediate dispositivi indossabili
- Author
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Pisano, Elisabetta, thesis supervisor: Lamberti, Claudio, Pisano, Elisabetta, and thesis supervisor: Lamberti, Claudio
39. Monitoraggio dell'attività motoria mediate dispositivi indossabili
- Author
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Pisano, Elisabetta, thesis supervisor: Lamberti, Claudio, Pisano, Elisabetta, and thesis supervisor: Lamberti, Claudio
40. HPLC-ESI-MS analysis of oral human fluids reveals that gingival crevicular fluid is the main source of oral thymosins beta(4) and beta(10).
- Author
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Inzitari R, Cabras T, Pisano E, Fanali C, Manconi B, Scarano E, Fiorita A, Paludetti G, Manni A, Nemolato S, Faa G, Castagnola M, and Messana I
- Subjects
- Adenoma, Pleomorphic pathology, Adenoma, Pleomorphic surgery, Adult, Chromatography, High Pressure Liquid, Female, Humans, Immunohistochemistry, Male, Reproducibility of Results, Salivary Gland Neoplasms pathology, Salivary Gland Neoplasms surgery, Sensitivity and Specificity, Spectrometry, Mass, Electrospray Ionization, Adenoma, Pleomorphic chemistry, Gingival Crevicular Fluid chemistry, Saliva chemistry, Salivary Gland Neoplasms chemistry, Thymosin analysis
- Abstract
Thymosin beta(4) (Tbeta(4)), its sulfoxide, and thymosin beta(10 )(Tbeta(10)) were detected in human saliva and identified by different strategies based on RP HPLC coupled to electrospray multidimensional IT MS. Tbeta(4 )was almost always detected in whole saliva, its sulfoxide sporadically, Tbeta(10) rarely. Tbeta(4) was undetectable in parotid saliva and less concentrated in submandibular/sublingual saliva than in whole saliva. Analysis of gingival crevicular fluid revealed high relative amounts of Tbeta(4), Tbeta(4) sulfoxide, and Tbeta(10) in all the samples. Tbeta(4) mean concentration was 200 times higher in crevicular fluid (20 micromol/L, N = 9) than in whole saliva (0.1 micromol/L, N = 9). Crevicular fluid concentration of Tbeta(4 )(ca. 5% represented by its sulfoxide) and beta(10 )significantly correlated (r = 0.856; N = 9), and their ratio was about 5. A significant correlation was also observed between Tbeta(4 )concentrations in whole saliva and gingival crevicular fluid (r = 0.738; N = 9). Immunohistochemical analysis of the major salivary glands showed that immunoreactivity for Tbeta(4) is restricted to ductal cells, with minor degree of focal positivity in some acinar cells. On the whole, results indicate that gingival sulcus is a main, although not the sole, source for oral Tbeta(4 )and Tbeta(10).
- Published
- 2009
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