Latest PUBLICATIONS

  • Glycolipids from sponges. Part 16.(1) discoside, a rare myo-inositol-containing glycolipid from the caribbean sponge Discodermia dissoluta.
    Publication Date: 01/10/2005, on Journal of natural products
    by Barbieri L, Costantino V, Fattorusso E, Mangoni A
    DOI: 10.1021/np050241q

    Discoside (1a), a glycolipid composed of 4,6-O-diacylated mannose attached to the 2-hydroxyl group of a myo-inositol unit, was isolated as a mixture of homologues from the marine sponge Discodermia dissoluta. The complete stereostructure of this new glycolipid was solved by interpretation of mass spectrometric and NMR data and CD analysis of degradation products.

  • Sporadic Creutzfeldt-Jakob disease: the extent of microglia activation is dependent on the biochemical type of PrPSc.
    Publication Date: 01/10/2005, on Journal of neuropathology and experimental neurology
    by Puoti G, Giaccone G, Mangieri M, Limido L, Fociani P, Zerbi P, Suardi S, Rossi G, Iussich S, Capobianco R, Di Fede G, Marcon G, Cotrufo R, Filippini G, Bugiani O, Tagliavini F
    DOI:

    In prion-related encephalopathies, microglial activation occurs early and is dependent on accumulation of disease-specific forms of the prion protein (PrPSc) and may play a role in nerve cell death. Previously, we found that different types of PrPSc (i.e. type 1 and type 2) coexisted in approximately 25% of patients with sporadic Creutzfeldt-Jakob disease (CJD); and a close relationship was detected between PrPSc type, the pattern of PrP immunoreactivity, and extent of spongiform degeneration. To investigate whether microglial reaction is related to the biochemical type and deposition pattern of PrPSc, we carried out a neuropathologic and biochemical study on 26 patients with sporadic CJD, including all possible genotypes at codon 129 of the prion protein gene. By quantitative analysis, we demonstrated that strong microglial activation was associated with type 1 PrPSc and diffuse PrP immunoreactivity, whereas type 2 PrPSc and focal PrP deposits were accompanied by mild microglia reaction. These findings support the view that the phenotypic heterogeneity of sporadic CJD is largely determined by the physicochemical properties of distinct PrPSc conformers.

  • Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer.
    Publication Date: 01/09/2005, on Molecular reproduction and development
    by Webster NL, Forni M, Bacci ML, Giovannoni R, Razzini R, Fantinati P, Zannoni A, Fusetti L, Dalprà L, Bianco MR, Papa M, Seren E, Sandrin MS, Mc Kenzie IF, Lavitrano M
    DOI: 10.1002/mrd.20316

    Multi-gene transgenic pigs would be of benefit for large animal models in medical, agricultural, and pharmaceutical applications; in particular for xenotransplantation, where extensive genetic manipulation of donor pigs is required to make them suitable for organ grafting to humans. We used the sperm mediated gene transfer (SMGT) method to produce with high efficiency multi-gene transgenic pigs using three genes coding for fluorescent proteins: enhanced blue (EBFP), green (EGFP), and red (DsRed2). All three fluorescent proteins were expressed in 171 out of 195 normally developed morula/blastocysts examined at day 6 post insemination (88%). Genomic DNA of 18 piglets born from two litters was screened by PCR, showing that all piglets were transgenic with at least one gene, 7/18 piglets were triple transgenic, 7/18 double transgenic, and 4/18 single transgenic. Fluorescence in situ hybridization (FISH) analysis revealed multiple sites of integration of the transgenes. RNA and protein expression was found in muscle, heart, liver, hair, and peripheral blood mononuclear cells (PBMCs). These results show that SMGT is an effective method for introducing multiple genes into pigs as shown by the simultaneous expression of three fluorescent proteins.

  • Leiomyosarcoma of the submandibular gland. Report of a case and review of the literature.
    Publication Date: 01/09/2005, on International journal of oral and maxillofacial surgery
    by Bucci T, Longo F, Mangone GM, Errico ME, Califano L
    DOI: 10.1016/j.ijom.2004.10.027

    This report describes the first case of primary leiomyosarcoma of the submandibular salivary glands and emphasizes the role of immunohistochemical study for a correct diagnosis of this tumour. In line with results of international literature, we associated surgery with radiotherapy and 2 years postoperatively there was no sign of recurrence.

  • Quadruplex-forming oligonucleotides as tools in anticancer therapy and aptamers design: energetic aspects.
    Publication Date: 01/09/2005, on Current medicinal chemistry. Anti-cancer agents
    by Petraccone L, Barone G, Giancola C
    DOI:

    Recent investigations on the G-quadruplex motif propose a new strategy for the making of antitumour drugs. Quadruplex-drug complexes have been suggested to inhibit telomerase activity; further, aptamers based on the quadruplex motif have been proved useful as tools aimed at binding and inhibiting particular proteins, thus serving as pharmaceutically active agents. However, the design of new aptamers is difficult because many factors affecting their activity and stability have not still been clarified. The knowledge of the energetics of quadruplex formation is a crucial point in view of their potential therapeutic utilization both as targets as well as therapeutic agents. In this review the energetic aspects of both quadruplex assembly and quadruplex-ligand interactions are discussed together with a summary of recent studies on physico-chemical properties in solution of quadruplex structures obtained from synthetic aptamers, including PNA-DNA chimeras.

  • A novel mutation in the RDS gene in an Italian family with pattern dystrophy.
    Publication Date: 01/08/2005, on The British journal of ophthalmology
    by Testa F, Marini V, Rossi S, Interlandi E, Nesti A, Rinaldi M, Varano M, Garré C, Simonelli F
    DOI: 10.1136/bjo.2004.064188

  • New unusual pregnane glycosides with antiproliferative activity from Solenostemma argel.
    Publication Date: 01/08/2005, on Steroids
    by Plaza A, Perrone A, Balestrieri ML, Felice F, Balestrieri C, Hamed AI, Pizza C, Piacente S
    DOI: 10.1016/j.steroids.2005.02.019

    Seven new 15-keto pregnane glycosides, namely Stemmosides E--K, were isolated from Solenostemma argel. Stemmosides E--J are characterized by the occurrence of an uncommon 14 beta proton configuration while stemmosides E and F possess in addition a rare enolic function in C-16. On the other hand, stemmosides G-J display an unusual C-17 alpha side chain. Their structures were established by ESI-MS and NMR experiments. Moreover, the effect of these compounds on the VEGF-induced in Kaposi's sarcoma cell proliferation was tested. Results indicated that all the compounds reduced the cell proliferation in a dose dependent manner.

  • Functional proteomics.
    Publication Date: 24/07/2005, on Clinica chimica acta; international journal of clinical chemistry
    by Monti M, Orrù S, Pagnozzi D, Pucci P
    DOI: 10.1016/j.cccn.2005.03.019

    With the increase in the number of genome sequencing projects, there is a concomitant exponential growth in the number of protein sequences whose function is still unknown. Functional proteomics constitutes an emerging research area in the proteomic field whose approaches are addressed towards two major targets: the elucidation of the biological function of unknown proteins and the definition of cellular mechanisms at the molecular level.

  • Pancreatic cancer-associated diabetes mellitus: an open field for proteomic applications.
    Publication Date: 24/07/2005, on Clinica chimica acta; international journal of clinical chemistry
    by Basso D, Greco E, Fogar P, Pucci P, Flagiello A, Baldo G, Giunco S, Valerio A, Navaglia F, Zambon CF, Pedrazzoli S, Plebani M
    DOI: 10.1016/j.cccn.2005.03.025

    Diabetes mellitus is associated with pancreatic cancer in more than 80% of the cases. Clinical, epidemiological, and experimental data indicate that pancreatic cancer causes diabetes mellitus by releasing soluble mediators which interfere with both beta-cell function and liver and muscle glucose metabolism.

  • Frontal dysfunction contributes to the genesis of hallucinations in non-demented Parkinsonian patients.
    Publication Date: 01/07/2005, on International journal of geriatric psychiatry
    by Grossi D, Trojano L, Pellecchia MT, Amboni M, Fragassi NA, Barone P
    DOI: 10.1002/gps.1339

    Hallucinations occur in patients with Parkinson's disease (PD) with reported prevalence ranging from 8% to 40%. Hallucinations are significantly associated with dementia in PD, but little is known about possible distinctive cognitive features of non-demented PD patients who develop hallucinations.

  • Direct interactions among Ret, GDNF and GFRalpha1 molecules reveal new insights into the assembly of a functional three-protein complex.
    Publication Date: 01/06/2005, on Cellular signalling
    by Amoresano A, Incoronato M, Monti G, Pucci P, de Franciscis V, Cerchia L
    DOI: 10.1016/j.cellsig.2004.10.012

    The glial-cell-line-derived neurotrophic factor (GDNF) ligand activates the Ret receptor through the assembly of a multiprotein complex, including the GDNF family receptor alpha1 (GFRalpha1) molecule. Given the neuroprotective role of GDNF, there is an obvious need to precisely identify the structural regions engaged in direct interactions between the three molecules. Here, we combined a functional approach for Ret activity (in PC12 cells) to cross-linking experiments followed by MS-MALDI to study the interactions among the purified extracellular region of the human Ret, GDNF and GFRalpha1 molecules. This procedure allowed us to identify distinct regions of Ret that are physically engaged in the interaction with GDNF and GFRalpha1. The lack of these regions in a recombinant Ret form results in the failure of both structural and functional binding of Ret to GFRalpha1/GDNF complex. Furthermore, a model for the assembly of a transducing-competent Ret complex is suggested.

  • Visual and spatial positive phenomena in the neglected hemifield--a case report.
    Publication Date: 01/06/2005, on Journal of neurology
    by Grossi D, Imperati F, Carbone G, Maiorino A, Angelillo V, Trojano L
    DOI: 10.1007/s00415-005-0724-0

  • tBid induces alterations of mitochondrial fatty acid oxidation flux by malonyl-CoA-independent inhibition of carnitine palmitoyltransferase-1.
    Publication Date: 01/06/2005, on Cell death and differentiation
    by Giordano A, Calvani M, Petillo O, Grippo P, Tuccillo F, Melone MA, Bonelli P, Calarco A, Peluso G
    DOI: 10.1038/sj.cdd.4401636

    Recent studies suggest a close relationship between cell metabolism and apoptosis. We have evaluated changes in lipid metabolism on permeabilized hepatocytes treated with truncated Bid (tBid) in the presence of caspase inhibitors and exogenous cytochrome c. The measurement of beta-oxidation flux by labeled palmitate demonstrates that tBid inhibits beta-oxidation, thereby resulting in the accumulation of palmitoyl-coenzyme A (CoA) and depletion of acetyl-carnitine and acylcarnitines, which is pathognomonic for inhibition of carnitine palmitoyltransferase-1 (CPT-1). We also show that tBid decreases CPT-1 activity by a mechanism independent of both malonyl-CoA, the key inhibitory molecule of CPT-1, and Bak and/or Bax, but dependent on cardiolipin decrease. Overexpression of Bcl-2, which is able to interact with CPT-1, counteracts the effects exerted by tBid on beta-oxidation. The unexpected role of tBid in the regulation of lipid beta-oxidation suggests a model in which tBid-induced metabolic decline leads to the accumulation of toxic lipid metabolites such as palmitoyl-CoA, which might become participants in the apoptotic pathway.

  • Autophagy is a prosurvival mechanism in cells expressing an autosomal dominant familial neurohypophyseal diabetes insipidus mutant vasopressin transgene.
    Publication Date: 01/06/2005, on FASEB journal : official publication of the Federation of American Societies for Experimental Biology
    by Castino R, Davies J, Beaucourt S, Isidoro C, Murphy D
    DOI: 10.1096/fj.04-3162fje

    Autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI) is a progressive, inherited neurodegenerative disorder that presents as polydipsia and polyuria as a consequence of a loss of secretion of the antidiuretic hormone vasopressin (VP) from posterior pituitary nerve terminals. VP gene mutations cause adFNDI. Rats expressing an adFNDI VP transgene (Cys67stop) show a neuronal pathology characterized by autophagic structures in the cell body. adFNDI has thus been added to the list of protein aggregation diseases, along with Alzheimer's, Parkinson's and Huntington's, which are associated with autophagy, a bulk process that delivers regions of cytosol to lysosomes for degradation. However, the role of autophagy in these diseases is unclear. To address the relationships between mutant protein accumulation, autophagy, cell survival, and cell death, we have developed a novel and tractable in vitro system. We have constructed adenoviral vectors (Ads) that express structural genes encoding either the Cys67stop mutant protein (Ad-VCAT-Cys67stop) or an epitope-tagged wild-type VP precursor (Ad-VCAT). After infection of mouse neuroblastoma Neuro2a cells, Ad-VCAT encoded material enters neurite processes and accumulates in terminals, while the Cys67stop protein is confined to enlarged vesicles in the cell body. Similar to the intracellular derangements seen in the Cys67stop rats, these structures are of ER origin, and colocalize with markers of autophagy. Neither Ad-VCAT-Cys67stop nor Ad-VCAT expression affected cell viability. However, inhibition of autophagy or lysosomal protein degradation, while having no effect on Ad-VCAT-expressing cells, significantly increased apoptotic cell death following Ad-VCAT-Cys67stop expression. These data suggest that activation of autophagy by the stress of the expression of an adFNDI mutant protein is a prosurvival mechanism.

  • Autophagy-dependent cell survival and cell death in an autosomal dominant familial neurohypophyseal diabetes insipidus in vitro model.
    Publication Date: 01/06/2005, on FASEB journal : official publication of the Federation of American Societies for Experimental Biology
    by Castino R, Isidoro C, Murphy D
    DOI: 10.1096/fj.04-3163fje

    Mutations in the human gene encoding the antidiuretic hormone vasopressin (VP) cause autosomal dominant familial neurohypophyseal diabetes insipidus (adFNDI), a rare inherited disorder that presents as polydipsia and polyuria as a consequence of a loss of secretion of VP from posterior pituitary nerve terminals. Work from our laboratories has shown that adFNDI, like other neurodegenerative diseases such as Alzheimer's, Parkinson's and Huntington's, is associated with autophagy. We have recently shown that the activation of autophagy in mouse neuroblastoma Neuro2a cells after adenoviral vector-mediated delivery of an adFNDI mutant VP transgene (Cys67stop) is a cell survival mechanism; its inhibition induces apoptosis. We now show that expression of Cys67stop sensitizes Neuro2a cells to the lethal effects of dopamine. This mode of cell death exhibits features typically associated with classical apoptosis. Yet inhibition of autophagy reversed these effects and rescued cell viability. We propose that autophagy-mediated cell death is a "two-hit" process: Following the cellular stress of the accumulation of a misfolded mutant protein, autophagy is prosurvival. However, a second insult triggers an autophagy-dependent apoptosis.