Latest PUBLICATIONS

  • Identification of trains of sleep sequences in adult rats.
    Publication Date: 15/02/2001, on Behavioural brain research
    by Piscopo S, Mandile P, Montagnese P, Cotugno M, Giuditta A, Vescia S
    DOI:

    In previous studies based on high resolution EEG analyses of the 7 h baseline session of 18 adult male Wistar rats [6,14], we have identified four sleep sequences initiating with slow wave sleep (SS) and terminating with waking (W) or paradoxical sleep (PS). Two of these sequences contained an intervening episode of transition sleep (TS). Several variables of these sequences (SS-->W, SS-->TS-->W, SS-->TS-->PS, and SS-->PS) were selectively correlated with the capacity of rats to learn a two-way active avoidance task the following day, and were differently distributed in fast learning, slow learning and non learning rats [21]. The temporal organization of different sleep components in sequences suggested that a comparable temporal organization might concern the different sleep sequences, albeit on a longer time scale. We have now used waking periods longer than 60 s to separate clusters of baseline sleep sequences (trains) in the same rats. Trains containing the same sleep sequence (homogeneous trains) have been distinguished from trains containing different sleep sequences (mixed trains). In addition, mixed trains including the SS-->TS-->W sequence (+TSW trains) have been separated from mixed trains lacking that sequence (-TSW trains). Mixed trains of the +TSW type were longest and most numerous, while homogeneous trains were shortest and least abundant. Mixed trains of the -TSW type displayed intermediate values. Several variables of sleep sequences and sleep components differed within mixed trains and among mixed and homogeneous trains. The data indicate that baseline sleep sequences aggregate in relatively long strings in a non random fashion. The mechanism of this association is discussed.

  • Molecular analysis of arterial stenosis in rat carotids.
    Publication Date: 01/02/2001, on Journal of cellular physiology
    by Forte A, Di Micco G, Galderisi U, Guarino FM, Cipollaro M, De Feo M, Gregorio R, Bianco MR, Vollono C, Esposito F, Berrino L, Angelini F, Renzulli A, Cotrufo M, Rossi F, Cascino A
    DOI: 10.1002/1097-4652(200002)186:2<307::AID-JCP1029>3.0.CO;2-I

    A new model of surgical injury for the induction and development of stenosis in common rat carotids is described. This model differs from balloon angioplasty or vein graft systems currently applied on animals to develop stenosis, since it involves the entire vessel wall layers and mimics the injury occurring during arterial grafts, endarterectomy or organ transplantation. At different times following arterial damage, the pattern of expression of genes already known to be involved in the proliferation, differentiation, and apoptosis of smooth muscle cells (c-myc, Angiotensin II receptor 1, Bcl-2 and Bax alpha), as well as of Rb and Rb2 genes, whose pattern of expression after arterial injury has not yet been reported, was analyzed by semi-quantitative reverse transcription-polymerase chain reaction technique. Histological and histochemical analysis on carotid sections shows the morphological changes which occurred 30 days after surgical injury in the vessel wall. Molecular and histological data demonstrate that this model of surgical injury induces neointimal proliferation in about 30% of rats. In about 70% of the remaining rats, it induces the processes responsible for negative remodelling, namely the significant accumulation of extracellular matrix and fibers and disorganization of arterial tunics. This model is therefore available for further studies on the expression of genes involved in the arterial stenotic process, as well as for testing drugs aimed at limiting this recurrent pathophysiological phenomenon.

  • A nucleotide insertion and frameshift cause albumin Kénitra, an extended and O-glycosylated mutant of human serum albumin with two additional disulfide bridges.
    Publication Date: 01/01/2001, on European journal of biochemistry
    by Minchiotti L, Campagnoli M, Rossi A, Cosulich ME, Monti M, Pucci P, Kragh-Hansen U, Granel B, Disdier P, Weiller PJ, Galliano M
    DOI:

    Albumin Kenitra is a new type of genetic variant of human serum albumin that has been found in two members of a family of Sephardic Jews from Kenitra (Morocco). The slow-migrating variant and the normal protein were isolated by anion-exchange chromatography and, after treatment with CNBr, the digests were analyzed by two-dimensional electrophoresis in a polyacrylamide gel. The CNBr peptides of the variant were purified by reverse-phase high performance liquid chromatography and submitted to sequence analysis. Albumin Kenitra is peculiar because it has an elongated polypeptide chain, 601 residues instead of 585, and its sequence is modified beginning from residue 575. DNA structural studies showed that the variant is caused by a single-base insertion, an adenine at nucleotide position 15 970 in the genomic sequence, which leads to a frameshift with the subsequent translation to the first termination codon of exon 15. Mass spectrometric analyses revealed that the four additional cysteine residues of the variant form two new S-S bridges and showed that albumin Kenitra is partially O-glycosylated by a monosialylated HexHexNAc structure. This oligosaccharide chain has been located to Thr596 by amino-acid sequence analysis of the tryptic fragment 592-597.

  • [Peripheral nervous system involvement in HCV-related mixed cryoglobulinemia]
    Publication Date: 01/01/2001, on Reumatismo
    by Migliaresi S, Di Iorio G, Ammendola A, Ambrosone L, Sanges G, Ugolini G, Sampaolo S, Bravaccio F, Tirri G
    DOI:

    In HCV-related mixed cryoglobulinemia (MC) a peripheral neuropathy (PN) may occur. To evaluate the prevalence and the characteristics of PN, 133 consecutive patients with HCV-MC (117 type II, 16 type III) were studied. Neurologic evaluation was performed according to the guidelines of Italian Group for the Study of Cryoglobulinemias, using a neurological disability score and a neurological symptom score. In 52/133 patients an electrophysiologic study (ENG) of ulnar, peroneal and sural nerves was performed. For 27/52 patients ENG data registered at different times (interval 12-96 months) were available. In 11 patients a sural nerve biopsy was obtained. An overt PN, mostly as sensory asymmetrical or symmetrical nerve impairment, was found in 107/133 patients (80.4%). ENG abnormalities-reduction or absence of sensory and sometimes of motor action potential, normal or slightly impaired nerve conduction velocity, consistent with axonal damage- were detected in 48/52 patients (92.3%). In 26 out of the 27 patients observed at different times an evolution of PN was found. Nerve biopsies showed a prevalent axonal damage, swollen endothelial cells in epi- and perineurial vessels and scarce mononuclear perivascular infiltrates. No leukocytoclastic vasculitis was observed. Immunoglobulins and complement in sub-perineurial vessel wall were detected. CONCLUSIONS: In HCV-MC a PN is frequent. It is mostly a sensory and progressively worsening axonopathy. Different mechanisms may be involved in the pathogenesis of this disorder and a direct role of HCV cannot be excluded.

  • Neurofilament homeostasis and motoneurone degeneration.
    Publication Date: 01/01/2001, on BioEssays : news and reviews in molecular, cellular and developmental biology
    by Perrone Capano C, Pernas-Alonso R, di Porzio U
    DOI: 10.1002/1521-1878(200101)23:1<24::AID-BIES1004>3.0.CO;2-H

    Neurofilament disorganisation is a hallmark of various neurodegenerative diseases. We review here current knowledge of neurofilament structure, gene expression and function. Neurofilament involvement in motoneurone neurological diseases is discussed in view of recent data from transgenic and spontaneous mouse mutants. In the mammalian neurone, the three neurofilament subunits are assembled into intermediate filaments as obligate heteropolymers. The subunits are expressed differentially during development and adult life according to the cell type and its physiological state. In addition to the well-established role of neurofilaments in the control of axonal calibre, there is increasing evidence that neurofilaments can interact with other cytoskeletal components and can modulate the axoplasmic flow. Although the extent to which neurofilament abnormalities contribute to the pathogenesis in human diseases remains unknown, emerging evidence suggests that disorganised neurofilaments can provoke degeneration and death of neurones. BioEssays 23:24-33, 2001.

  • Cervical ganglioneuroma: report of a case.
    Publication Date: 01/01/2001, on Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery
    by Califano L, Zupi A, Mangone GM, Long F
    DOI: 10.1067/mhn.2001.111370

  • Intraosseous mucoepidermoid carcinoma. Report of a long-term evolution case.
    Publication Date: 01/01/2001, on Oral oncology
    by Maremonti P, Califano L, Mangone GM, Zupi A, Guida C
    DOI:

    Primary central mucoepidermoid carcinoma (CMC) of the jaws accounts only for 2-3% of all mucoepidermoid carcinomas reported. Bhaskar in 1963 first analysed the criteria for his central origin, histology and pathogenesis. The authors report a long-term evolution case of CMC of the mandible with peculiar clinical features observed at the Department of Maxillo-Facial Surgery of the School of Medicine and Surgery of the "Federico II" University of Naples (Naples, Italy) examining histopathologic and clinical features and problems related to the treatment.

  • Characterization of five new low-molecular-mass trypsin inhibitors from white mustard (Sinapis alba L.) seed.
    Publication Date: 01/11/2000, on European journal of biochemistry
    by Ruoppolo M, Amoresano A, Pucci P, Pascarella S, Polticelli F, Trovato M, Menegatti E, Ascenzi P
    DOI:

    Five new low-molecular-mass trypsin inhibitors belonging to the RTI/MTI-2 family were identified from white mustard (Sinapis alba L. ; MTI-2) seed. Purified MTI-2 consisted of a peptide mixture, displaying Ile or Arg at position 43, Trp or kynurenine (Kyn) at position 44, and C-terminal ragged ends. The occurrence of Ile or Arg at position 43 suggested that MTI-2 inhibitors originated from different genes. The presence of 5-oxo-proline (pyroglutamic acid; 5-oxoPro1) and Kyn44 reflected post-translational processing of the serine proteinase inhibitor. MTI-2 showed approximately 70% amino-acid identity with low-molecular-mass trypsin inhibitors isolated from oil rape (Brassica napus var. oleifera; RTI-III) seed and with serine proteinase inhibitors mapped in Arabidopsis thaliana chromosome II (ATTI). Furthermore, MTI-2 was homologous to brazzein, the sweet-tasting protein from Pentadiplandra brazzeana Baillon fruit ( approximately 30% amino-acid identity). Although snake-venom toxins showed a low amino-acid identity (< 20%) with MTI-2, RTI-III, and ATTI, some structurally relevant residues were conserved. The disulfide bridge pattern of MTI-2 (Cys5-Cys27, Cys18-Cys31, Cys42-Cys52, and Cys54-Cys57) corresponded to that of RTI-III and of snake-venom toxins, being different from that of brazzein. Therefore, protein similarity might be attributable to the three-dimensional arrangement rather than to the amino-acid sequence. Values of Ka for MTI-2 binding to bovine beta-trypsin (trypsin) and bovine alpha-chymotrypsin were 6.3 x 109 M-1 and 2.0 x 106 M-1, respectively, at pH 8.0 and 21.0 degrees C. Moreover, values of kon for MTI-2 binding to trypsin and of koff for the dissociation of the serine proteinase:inhibitor complex were 5.6 x 105 M-1.s-1 and 8.9 x 10-5 M-1.s-1, respectively, at pH 8.0 and 21.0 degrees C. Despite the heterogeneity of the purified inhibitor peptide mixture, the inhibition properties of the different MTI-2 inhibitors were indistinguishable.

  • Increased expression of IGF-binding protein-5 in Duchenne muscular dystrophy (DMD) fibroblasts correlates with the fibroblast-induced downregulation of DMD myoblast growth: an in vitro analysis.
    Publication Date: 01/10/2000, on Journal of cellular physiology
    by Melone MA, Peluso G, Galderisi U, Petillo O, Cotrufo R
    DOI: 10.1002/1097-4652(200010)185:1<143::AID-JCP14>3.0.CO;2-U

    In DMD the progressive loss of muscle ability and concomitant increasing fibrosis might originate from, besides other causes, the fibroblast paracrine inhibition of satellite cell "growth." In this study we report that in myoblast/fibroblast coculture experiments, the presence of DMD fibroblasts negatively interfered with DMD myoblast growth to an extent directly proportional to the percentage of DMD fibroblasts present in the mixed-cell cultures. Moreover, the observation that media conditioned with proliferating DMD fibroblasts inhibited the growth of DMD myoblasts more seriously than did control fibroblast-conditioned media suggested a paracrine effect by diffusible factors. IGF-binding proteins could act as such diffusible factors; in fact, IGFBP-5 transcript increased threefold in DMD fibroblasts proliferating in DMD muscle extracts, whereas IGFBP-3 mRNA decreased. In addition, high levels of IGFBP-5 protein were detected in DMD fibroblast-conditioned media. In neutralizing IGFBP-5 in DMD fibroblast-conditioned media by means of specific antibodies, or inhibiting IGFBP-5 gene expression in DMD fibroblasts by means of oligo antisense, the fibroblast-conditioned media lost inhibitory power over DMD myoblast proliferation.

  • Spatial cognition in children. II. Visuospatial and constructional skills in developmental reading disability.
    Publication Date: 01/09/2000, on Brain & development
    by Del Giudice E, Trojano L, Fragassi NA, Posteraro S, Crisanti AF, Tanzarella P, Marino A, Grossi D
    DOI:

    Cognitive models for developmental dyslexia are nowadays centered on the hypothesis of a specific deficit within the phonologic module of the language system. To ascertain whether defects of spatial cognition are associated with developmental reading disability, we investigated a sample of 43 school children (aged 8-9 years) found to be reading impaired during a wide screening survey for developmental dyslexia in the province of Naples, Italy. After one year all children were tested again and only 9/43 still presented reading impairment, while the remaining had achieved a variable range of spontaneous recovery. A detailed analysis was performed on all children to characterize their cognitive performances using on one hand classical conventional tests for constructional praxis, visuospatial cognition, and visuospatial memory and on the other a specific neuropsychological battery for constructional disorders. The results of our study demonstrated that children with long-lasting reading impairment exhibited normal performances on spatial cognition tasks. Moreover, one single child was found with relevant visuospatial deficits pointing to the possible existence of a visuospatial subtype for developmental dyslexia.

  • Spatial cognition in children. I. Development of drawing-related (visuospatial and constructional) abilities in preschool and early school years.
    Publication Date: 01/09/2000, on Brain & development
    by Del Giudice E, Grossi D, Angelini R, Crisanti AF, Latte F, Fragassi NA, Trojano L
    DOI:

    The present study aimed to investigate the acquisition of visuospatial and graphomotor capacities during the pre-school and early schooling years in order to follow the normal development of drawing-related abilities and spatial cognition. Eighty children aged 3-5 years, divided in four subgroups each different for a 6-month period, and 80 children aged 8-9 years were administered a neuropsychological battery for visuospatial and visuoconstructional analysis. The battery explored five cognitive domains: visual scanning, visuospatial perceptual and representational abilities, visuomotor control and graphomotor skills. Results showed that the total scores significantly improved in each group of children with respect to the previous one, but the pattern of skill acquisition was not homogeneous. We observed a gradient from explorative and visuomotor to perceptive, representational and graphomotor abilities. Explorative and visuomotor abilities were almost mature at a time when visuoperceptual capacities began to develop. On the contrary, at that time we found very low performances at representational and constructional tasks. Our findings could suggest that constructional abilities need both perceptual and representational competences to develop properly.

  • The lysosomal protease cathepsin D is efficiently sorted to and secreted from regulated secretory compartments in the rat basophilic/mast cell line RBL.
    Publication Date: 01/09/2000, on Journal of cell science
    by Dragonetti A, Baldassarre M, Castino R, Démoz M, Luini A, Buccione R, Isidoro C
    DOI:

    Basophils and mast cells contain a peculiar class of inflammatory granules that discharge their content upon antigen-mediated crosslinking of IgE-membrane receptors. The pathways for granule biogenesis and exocytosis in these cells are still largely obscure. In this study we employed the rat basophilic leukemia (RBL)/mast cell line to verify the hypothesis that inflammatory granules share common bioactive molecules and functional properties with lysosomes. We demonstrate that inflammatory granules, as identified by the monoclonal 5G10 antibody (which recognises an integral membrane protein) or by Toluidine Blue staining, have an intralumenal acidic pH, possess lysosomal enzymes and are accessible by fluid-phase and membrane endocytosis markers. In addition, we studied the targeting, subcellular localisation and regulated secretion of the lysosomal aspartic protease cathepsin D (CD) as affected by IgE receptor stimulation in order to obtain information on the pathways for granule biogenesis and exocytosis. Stimulation with DNP-BSA of specific IgE-primed RBL cells led to a prompt release of processed forms of CD, along with other mature lysosomal hydrolases. This release could be prevented by addition of EGTA, indicating that it was dependent on extracellular calcium influx. Antigen stimulation also induced exocytosis of immature CD forms accumulated by ammonium chloride, suggesting the existence of an intermediate station in the pathway for granule biogenesis still sensitive to regulated exocytosis. The targeting of molecules to secretory granules may occur via either a mannose-6-phosphate-dependent or mannose-6-phosphate-independent pathway. We conclude that endosomes and lysosomes in basophils/mast cells can act as regulated secretory granules or actually identify with them.

  • Regulation of the biosynthesis of acyl analogs of platelet-activating factor by purinergic agonist in endothlial cells.
    Publication Date: 11/08/2000, on FEBS letters
    by Balestrieri ML, Lee T
    DOI:

    We have previously shown that platelet-activating factor (PAF)-dependent transacetylase (TA) contains three catalytic activities, namely PAF: lysophospholipid TA (TAL), PAF: sphingosine TA (TAs) and PAF acetylhydrolase. It serves as a modifier of PAF actions by producing different lipid signal molecules. The TAL activity is involved in the biosynthesis of acyl analogs of PAF (acyl-PAF, 1-acyl-2-acetyl-sn-glycero-3-phosphocholine, acylacetyl-GPC) in agonist-stimulated endothelial cells. In the present investigation, we have studied the mechanism(s) by which the TA activity is regulated in ATP-treated endothelial cells. We have demonstrated that ATP, and thiol-modifying agents with ATP, specifically regulate only the TAL part of the TA activities.

  • Creutzfeldt-Jakob disease with a novel four extra-repeat insertional mutation in the PrP gene.
    Publication Date: 08/08/2000, on Neurology
    by Rossi G, Giaccone G, Giampaolo L, Iussich S, Puoti G, Frigo M, Cavaletti G, Frattola L, Bugiani O, Tagliavini F
    DOI:

    To investigate the role of a short insertional mutation in the prion protein (PrP) gene (PRNP) in prion disease pathogenesis.

  • Mass spectrometry study of ecto-5'-nucleotidase from bull seminal plasma.
    Publication Date: 01/08/2000, on European journal of biochemistry
    by Fini C, Amoresano A, Andolfo A, D'auria S, Floridi A, Paolini S, Pucci P
    DOI:

    The structure of ecto-5'-nucleotidase from bull seminal plasma, containing a glycosyl-phosphatidylinositol anchor, was studied using mass spectrometry. MALDI-MS analysis of intact protein indicated a mass of 65 568.2 Da for the monomeric form, and it also showed a heterogeneous population of glycoforms with the glycosidic moiety accounting for approximately 6000 Da. MALDI-MS analysis showed that Asn53, Asn311, Asn333 and Asn403 were four sites of N-glycosylation. GC-MS analysis provided information on the glycosidic structures linked to the four asparagines. Asn53, Asn311 and Asn333 were linked to high-mannose saccharide chains, whereas the glycan chains linked to Asn403 contained a heterogeneous mixture of oligosaccharides, the high-mannose type structure being the most abundant and hybrid or complex type glycans being minor components. By combining enzymatic and/or chemical hydrolysis with GC-MS analysis, detailed characterization of the glycosyl-phpsphatidylinositol anchor was obtained. MALDI spectral analysis indicated that the glycosyl-phosphatidylinositol core contained EtN(P)Man3GlcNH2-myo-inositol(P)-glycerol, principally modified by stearoyl and palmitoyl residues or by stearoyl and myristoyl residues to a minor extent. Moreover, 1-palmitoylglycerol and 1-stearoylglycerol outweighed 2-palmitoylglycerol and 2-stearoylglycerol. The combination of chemical and enzymatic digestions of the protein with the mass spectral analysis yielded a complete pattern of S-S bridges. The protein does not contain free thiols and its eight cysteines are linked by intramolecular disulfide bonds, the pairs being: Cys51-Cys57, Cys353-Cys358, Cys365-Cys387 and Cys476-Cys479. This work resolves details of the structure of ecto-5'-nucleotidase, with particular regard to the localization and composition of the glycidic moiety, number and localization of the disulfide bridges and characterization of the glycosyl-phosphatidylinositol anchor.