P Martelli

Università degli Studi di Siena, Siena, Tuscany, Italy

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Publications (61)100.27 Total impact

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    ABSTRACT: Abstract Sections of Ammi visnaga (L.) Lam. ripe fruits were examined by U.V. microscopy under U.V. 365 nm light and under mixed U.V. 365 nm and visible light. The examination revealed a substance with blue autofluorescence inside the primary rib channels; a blue colour was shown also by the endosperm, whilst vittae were empty. In sections extracted with a selective method for the furanochromones khellin and visnagin, the substance filling primary rib channels was completely dissolved, and the colour of endosperm faded. The extract was analysed by high-performance liquid chromatography: the solubilized materials were composed almost exclusively of khellin and visnagin. These data are in contrast with the previous consensus which considered coumarins and coumarin-like compounds to be located in the vittae of Umbelliferae fruits, and never present in endosperms, whilst primary rib channels were thought empty in ripe fruits.
    09/2008; 25(3):137-144.
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    ABSTRACT: Meningococcal disease is a global problem. Multivalent (A, C, Y, W135) conjugate vaccines have been developed and licensed; however, an effective vaccine against serogroup B has not yet become available. Outer membrane vesicle (OMV) vaccines have been used to disrupt serogroup B epidemics and outbreaks. Postgenomic technologies have been useful in aiding the discovery of new protein vaccine candidates. Moreover, proteomic technologies enable large-scale identification of membrane and surface-associated proteins, and provide suitable methods to characterize and standardize the antigen composition of OMV-based vaccines.
    Expert Review of Proteomics 11/2007; 4(5):667-77. · 3.90 Impact Factor
  • International Journal of Ecodynamics. 01/2006; 1(3):266-283.
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    ABSTRACT: Phospholipase activity, one of Helicobacter pylori pathogenicity factors, has not been investigated enough, so far, although it may induce a remarkable damage to the gastric mucosa. In the present work, we have compared the whole phospholipase activity of H. pylori strains isolated from patients with gastric carcinoma with that of strains isolated from dyspeptic patients without gastric carcinoma. We measured the phospholipase activity of one distinct H. pylori colony isolated from each of 10 patients with gastric carcinoma and 10 controls, dyspeptic patients without endoscopic and histological signs of gastric carcinoma. We also determined the phospholipase activity of 20 additional strains isolated from different areas of neoplastic and non-neoplastic tissue of two patients with gastric carcinoma, the cagA and vacA positive G27 and 328 wild strains and their respective vacA and cagA negative isogenic mutants. The whole phospholipase activity of strains was determined by measuring the release of (14)C-labeled palmitic acid from the radioactive l-3-phosphatidylcholine, 1,2-di[1-(14)C]palmiloyl substrate; results were expressed in pmol of palmitic acid per mg of protein. H. pylori strains isolated from patients with gastric carcinoma had levels of phospholipase activity significantly higher than those of strains isolated from controls (99.37 [S.D. 40.45] versus 34.46 [S.D. 16.46], P<0.001). In patients with gastric carcinoma, the mean phospholipase activity of strains isolated from neoplastic tissue was similar to that of strains isolated from non-neoplastic tissues (123.02 [S.D. 44.36] and 115.77 [S.D. 81.48], respectively. Interruption of cagA gene caused a ca. 20% reduction of phospholipase activity (36.38 versus 45.22 of the wild strain); that of vacA caused no reduction of phospholipase activity (26.53 and 25.37 of the wild strain). The infection by H. pylori strains that produce high levels of phospholipase may increase the risk of developing gastric carcinoma. We hypothesise that indirect products of phospholipase activity, such as prostaglandins, leukotrienes and lysophospholipids, may mediate carcinogenesis.
    Digestive and Liver Disease 04/2005; 37(4):232-9. · 3.16 Impact Factor
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    ABSTRACT: Rosacea and chronic urticaria are two common skin disorders existing in idiopathic forms. A role of Helicobacter pylori bacterium infection in the aetiopathogenesis of rosacea or chronic urticaria has been suggested although still controversial. The aim of the present study was to establish a relationship between H. pylori infection and rosacea chronic urticaria by means of an immunoproteomic investigation. We analyzed immunoglobulin A (IgA)-, IgG-, and IgE-mediated immune-responses against H. pylori antigens and we identified some bacterial immunoresponsive proteins. A general IgA- and IgE-mediated immune response against antioxidative bacterial proteins was observed. A correlation between the bacterial occurrence and skin diseases pathogenesis is discussed.
    PROTEOMICS 03/2005; 5(3):777-87. · 4.13 Impact Factor
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    ABSTRACT: Neisseria meningitidis is an encapsulated Gram-negative bacterium responsible for significant morbidity and mortality worldwide. Meningococci are opportunistic pathogens, carried in the nasopharynx of approximately 10% of asymptomatic adults. Occasionally they enter the bloodstream to cause septicaemia and meningitis. Meningococci are classified into serogroups on the basis of polysaccharide capsule diversity, and serogroup A strains have caused major epidemics mainly in the developing world. Here we describe a two-dimensional gel electrophoresis protein map of the serogroup A strain Z4970, a clinical isolate classified as ancestral to several pandemic waves. To our knowledge this is the first systematically annotated proteomic map for N. meningitidis. Total protein samples from bacteria grown on GC-agar were electrophoretically separated and protein species were identified by matrix-assisted laser desorption/ionization time of flight spectrometry. We identified the products of 273 genes, covering several functional classes, including 94 proteins so far considered as hypothetical. We also describe several protein species encoded by genes reported by DNA microarray studies as being regulated in physiological conditions which are relevant to natural meningococcal pathogenicity. Since menA differs from other serogroups by having a fairly stable clonal population structure (i.e. with a low degree of variability), we envisaged comparative mapping as a useful tool for microevolution studies, in conjunction with established genotyping methods. As a proof of principle, we performed a comparative analysis on the B subunit of the meningococcal transferrin receptor, a vaccine candidate encoded by the tbpB gene, and a known marker of population diversity in meningococci. The results show that TbpB spot pattern variation observed in the maps of nine clinical isolates from diverse epidemic spreads, fits previous analyses based on allelic variations of the tbpB gene.
    PROTEOMICS 11/2004; 4(10):2893-926. · 4.13 Impact Factor
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    ABSTRACT: Plasma protein adsorption patterns on surfaces may give vital information to evaluate biocompatibility of biomaterials designed for direct blood-contacting applications or tissue integration. Adsorption of human serum proteins on four different types of biomaterials (glass, aminosilanized glass, hyaluronan and sulfated hyaluronan) was analyzed by two-dimensional electrophoresis. Desorption of proteins from the surfaces was first classically achieved by sodium dodecyl sulfate (SDS) elution. We introduced a second elution step (by use of isoelectric focusing (IEF) sample buffer consisting of urea, 3-[(3-cholamidopropyl)dimethylammonio]-1-propansulfonate, and dithioerythritol) which allows more stringent elution conditions and is a tool to evaluate the protein adsorption strength to biomaterials. Moreover, the two-step elution may discriminate between irreversible and reversible adsorption of plasma proteins for biomaterials, thus helping to elucidate the structure of protein multilayers which form a complex system at the surfaces. The IEF sample buffer proved not to alter the biomaterial structure and integrity. Hydrophobic bonds resulted to be the main strength driving protein adsorption onto our biomaterials. Apolipoproteins were the most important proteins interacting with the surfaces suggesting that high-density lipoprotein (HDL) particles could play a role in biocompatibility due to their beneficial effects on endothelial cells.
    Electrophoresis 08/2004; 25(14):2413-24. · 3.26 Impact Factor
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    ABSTRACT: The cytotoxin-associated protein CagA is a Helicobacter pylori immunodominant antigen whose gene resides in the cag pathogenicity island. Our purpose was to determine if the disruption or deletion of cagA gene could have an effect on the expression of other proteins at the proteome level. We analyzed two H. pylori strains, 328 and G27 wild-type, bearing the cag pathogenicity island, and their respective isogenic cagA(-) mutants. The proteomes of two H. pylori strains (328 and its isogenic mutant SPM328_DeltacagA) were resolved by two-dimensional electrophoresis and the digitalized images obtained were analysed both quantitatively and qualitatively. Peculiar spots of each strain were identified by mass spectrometry or by Western blotting. The comparison between the proteome expression of an H. pylori cagA(+) strain and an isogenic mutant strain where the cagA gene was disrupted showed that, as well as the lack of expression of CagA, both flagellin A and flagellin B expressions were significantly decreased. The cagA(-) isogenic mutant was nonmotile. G27_DeltacagA, in which CagA was inactivated by gene deletion, was nonmotile as well respecting to motile G27 wild-type strain. Moreover, reintroduction of cagA in G27_DeltacagA restored motility. Our results suggest that CagA could quantitatively influence flaA and flaB transcription or their subsequent translation and/or correct folding.
    Helicobacter 07/2004; 9(3):185-93. · 3.51 Impact Factor
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    ABSTRACT: This paper describes the project "Biotech a Scuola" ("Biotech at School"), financed by the Italian Ministry of Education within the SeT program (Special Project for Scientific-Technological Education). The project involved the University of Siena, five senior and junior secondary schools in the Siena area, and a private company. Twenty-three teachers from diverse fields and 318 students from 15 classes were involved. The aim of the project was to improve scientific-technological teaching by providing schools with the support and materials necessary to understand some fundamental aspects of biotechnology. With this project we propose a model of close cooperation among various educational sectors with the goal of teaching junior and senior high school students some of the theory and practice of modern biotechnology.
    Biochemistry and Molecular Biology Education 03/2004; 32(2):78-83. · 0.70 Impact Factor
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    ABSTRACT: Saccharomyces cerevisiae is the optimal eukaryotic model system to study mammalian biological responses. At the same time Saccharomyces cerevisiae is also widely utilized as a biotechnological tool in the food industry. Enological Saccharomyces cerevisiae strains have been so far routinely analyzed for their microbiological aspects. Nevertheless, wine yeasts are gaining an increasing interest in the last years since they strongly affect both the vinification process and the organoleptic properties of the final product wine. The protein repertoire is responsible of such features and, consequently, 2D-PAGE can be an useful tool to evaluate and select optimal wine yeast strains. We present here the first proteomic map of a wild-type wine Saccharomyces cerevisiae strain selected for the guided fermentation of very high quality wines.
    The Italian journal of biochemistry 01/2004; 52(4):145-53.
  • The Italian journal of biochemistry 01/2004; 52(4):125-9.
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    ABSTRACT: We report a study on the adaptive response of a wild-type wine Saccharomyces cerevisiae strain, isolated from natural spontaneous grape must, to mild and progressive physiological stresses due to fermentation. We observed by two-dimensional electrophoresis how the yeast proteome changes during glucose exhaustion, before the cell enters its complete stationary phase. On the basis of their identification, the proteins representing the S. cerevisiae proteomic response to fermentation stresses were divided into three classes: repressed proteins, induced proteins and autoproteolysed proteins. In an overall view, the proteome adaptation of S. cerevisiae at the time of glucose exhaustion seems to be directed mainly against the effects of ethanol, causing both hyperosmolarity and oxidative responses. Stress-induced autoproteolysis is directed mainly towards specific isoforms of glycolytic enzymes. Through the use of a wild-type S. cerevisiae strain and PMSF, a specific inhibitor of vacuolar proteinase B, we could also distinguish the specific contributions of the vacuole and the proteasome to the autoproteolytic process.
    Biochemical Journal 03/2003; 370(Pt 1):35-46. · 4.65 Impact Factor
  • Digestive and Liver Disease 06/2002; 34. · 3.16 Impact Factor
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    ABSTRACT: We have previously shown that human plasma atrial alpha-natriuretic peptide (alpha-hANP) sequestering is a protective phenomenon against amyloid aggregation. In the present work, the possible role of lipoproteins as alpha-hANP binding factors has been investigated in vitro using an experimental model, developed in our laboratory, that allows to work at physiological concentrations. This approach consists of gel filtration on Sephacryl S-300 HR of big alpha-[(125)I]hANP generated in phosphate buffered saline or in human normal plasma supplemented or not with lipoproteins. The results of these experiments indicate that high density lipoproteins (HDL) are responsible for the ANP binding phenomenon observed in vitro, while low density lipoproteins and very low density lipoproteins do not directly interact with ANP. Moreover, the HDL remodeling process occurring in vitro has been analyzed during plasma incubation by monitoring the redistribution of lipids and apolipoproteins among the HDL subclasses. The changes in HDL size and composition observed in incubated plasma were compared with the redistribution of endogenous and labeled big ANP. The obtained results revealed that both tend to follow the molecular rearrangement in plasma of apolipoprotein A-I containing particles and suggested that, among HDL species, the small particles are mainly involved in the ANP binding phenomenon. This hypothesis was further demonstrated by ligand blotting experiments that confirmed the existence of differences in the ability of HDL particles to bind alpha-[(125)I]hANP.
    Biochimica et Biophysica Acta 06/2001; 1536(2-3):123-32. · 4.66 Impact Factor
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    ABSTRACT: We recently studied the protein composition of a Saccharomyces cerevisiae wine yeast strain (K310) of enological interest. About 2500 spots of 8—250 kDa observed molecular mass were resolved by two-dimensional gel electrophoresis. Experimental molecular masses and isoelectric points were calculated for most of them. Twenty-seven proteins were subjected to Edman microsequencing. N-terminal sequences of 12/27 proteins were determined, whereas internal sequences of 6/27 proteins were obtained following in situ proteolysis. Comparison between the experimental data and those reported in the SWISS-PROT database revealed some differences between genotypic and phenotypic sequences. These are indicative of the changes a protein can undergo with respect to the primary structure coded by the genomic DNA. Our results highlight the need to complement genomic analysis with detailed proteomics in order to refine the vast amount of information provided by DNA sequencing and to find an exact correlation between genome and proteome.
    Electrophoresis 12/2000; 21(17):3717 - 3723. · 3.26 Impact Factor
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    ABSTRACT: We recently studied the protein composition of a Saccharomyces cerevisiae wine yeast strain (K310) of enological interest. About 2,500 spots of 8-250 kDa observed molecular mass were resolved by two-dimensional gel electrophoresis. Experimental molecular masses and isoelectric points were calculated for most of them. Twenty-seven proteins were subjected to Edman microsequencing. N-terminal sequences of 12/27 proteins were determined, whereas internal sequences of 6/27 proteins were obtained following in situ proteolysis. Comparison between the experimental data and those reported in the SWISS-PROT database revealed some differences between genotypic and phenotypic sequences. These are indicative of the changes a protein can undergo with respect to the primary structure coded by the genomic DNA. Our results highlight the need to complement genomic analysis with detailed proteomics in order to refine the vast amount of information provided by DNA sequencing and to find an exact correlation between genome and proteome.
    Electrophoresis 12/2000; 21(17):3717-23. · 3.26 Impact Factor
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    ABSTRACT: The existence of a Surface Tension Lower Substance (STLS) in the lung and the Eustachian tube was established a few decades ago. The histological and physiological characteristics of the epithelium of the whole respiratory tract, from the nasal cavities to rhinopharynx and bronchiolar segments, induced the Authors to search for such a similar substance also in nasal secretion. To this purpose nasal secretions were studied using High Pressure Liquid Chromatography (HPLC) to determine the presence of phospholipids. In addition, ultrastructural examination of the nasal mucosa was performed looking for lamellar body-like structures of 2nd degree pulmonary cells.
    Acta otorhinolaryngologica Italica: organo ufficiale della Società italiana di otorinolaringologia e chirurgia cervico-facciale 05/1997; 17(2):93-6. · 0.79 Impact Factor
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    ABSTRACT: Although Ras and Rap1 share interaction with common candidate effector proteins, Rap1 lacks the transforming activity exhibited by Ras proteins. It has been speculated that Rap antagonizes Ras transformation through the formation of nonproductive complexes with critical Ras effector targets. To understand further the distinct biological functions of these two closely related proteins, we searched for Rap1b-binding proteins by yeast two-hybrid screening. We identified multiple clones that encode the COOH-terminal sequences of a protein that shares sequence identity with RalGDS and RGL, which we have designated RGL2. A 158-amino acid COOH-terminal fragment of RGL2 (RGL2 C-158) bound to Ras superfamily proteins which shared identical effector domain sequences with Rap1 (Ha-Ras, R-Ras, and TC21). RGL2 C-158 binding was impaired by effector domain mutations in Rap1b and Ha-Ras. Furthermore, RGL2 C-158 bound exclusively to the GTP-, but not the GDP-bound form of Ha-Ras. Finally, coexpression of RGL2 C-158 impaired oncogenic Ras activation of transcription from a Ras-responsive promoter element and focus-forming activity in NIH 3T3 cells. We conclude that RGL2 may be an effector for Ras and/or Rap proteins.
    Journal of Biological Chemistry 12/1996; 271(47):29903-8. · 4.65 Impact Factor
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    ABSTRACT: Khellin, a furochromone structurally related to furocoumarins, is a phototherapeutic agent used against psoriasis and vitiligo. This paper reports that pretreatment of HeLa cells with pertussis toxin reverses both the inhibition of NaF-stimulated adenylyl cyclase activity and the stimulation of GTPase by khellin alone and plus UVA light, as previously reported. Our results demonstrate, for the first time, that khellin is able to elicit an intracellular response by Gi alpha protein.
    Biochimica et Biophysica Acta 12/1996; 1314(1-2):105-8. · 4.66 Impact Factor
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    ABSTRACT: Until recently, the therapeutic effects of furocoumarins and furochromones plus UV-A light were thought to be due to their ability to form photoadducts with DNA in the cell nuclei; now it appears that membrane effector systems may be involved as targets. Here we show that in HeLa cells khellin at 1 and 5 microM final concentration, in combination with UV-A light, inhibits NaF-stimulated adenylyl cyclase activity and Pertussis Toxin (PT)-catalyzed ADP-ribosylation of alpha-subunits of inhibitory guanine nucleotide regulatory proteins (Gi) and increases GTPase activity. In the same experimental conditions, 8-methoxypsoralen (8-MOP), either alone or plus UV-A, does not affect adenylyl cyclase and GTPase activities. Our results suggest that in HeLa cells, through an interaction with a receptor and the mediation of Gi proteins, the adenylyl cyclase system is a target for khellin but not for 8-MOP.
    Biochimica et Biophysica Acta 12/1995; 1269(2):162-6. · 4.66 Impact Factor