Publications by authors named "Virginia Carbone"

24 Publications

  • Page 1 of 1

Grape Canes from Typical Cultivars of Campania (Southern Italy) as a Source of High-Value Bioactive Compounds: Phenolic Profile, Antioxidant and Antimicrobial Activities.

Molecules 2021 May 7;26(9). Epub 2021 May 7.

Research Institute on Terrestrial Ecosystems, National Research Council of Italy, Via Pietro Castellino 111, 80131 Naples, Italy.

The purpose of the current study was to determine the phenolic composition, antioxidant, and antimicrobial activities in grape cane extracts from typical cultivars of Southern Italy. Aqueous extracts at different pHs (1-13) were prepared from "Aglianico", "Fiano", and "Greco" grape canes. The results demonstrated that an alkaline pH (13.00) produced the best polyphenol-rich extracts, as the total phenolic content was more than double when compared to the respective extracts prepared at pH 1.00. "Greco" grape canes gave the highest quantity of phenolic compounds at each pH, ranging from 42.7 ± 0.4 to 104.3 ± 3.0 mg Gallic Acid Equivalents (GAE)/g Dry Extract (DE) from pH 1.00 to 13.00. The Radical Scavenging Activity (RSA) and the Ferric Reducing Antioxidant Power (FRAP) were measured. The highest antioxidant activity was showed by "Greco" extract at pH 7.00. Seventy-five compounds were identified in the extracts by HPLC-MS with six of them described for the first time in grape canes. Procyanidins were highly abundant in extracts at pH 7.00, whereas stilbenoids were the most represented compounds at pH 13.00. Very strong antiviral activity against herpes simplex viruses was recorded for the extracts at pH 7.00 and 13.00 that were active in the early stages of infection by acting directly against the viral particles. The overall results suggest that grape canes, currently underutilized, can be usefully valorised by providing active extracts to use as antioxidant and antiviral agents.
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http://dx.doi.org/10.3390/molecules26092746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8125794PMC
May 2021

Unveiling Kiwifruit Metabolite and Protein Changes in the Course of Postharvest Cold Storage.

Front Plant Sci 2019 4;10:71. Epub 2019 Feb 4.

Proteomics & Mass Spectrometry Laboratory, Istituto per il Sistema Produzione Animale In Ambiente Mediterraneo, National Research Council, Naples, Italy.

cv. Hayward fruit is renowned for its micro- and macronutrients, which vary in their levels during berry physiological development and postharvest processing. In this context, we have recently described metabolic pathways/molecular effectors in fruit outer endocarp characterizing the different stages of berry physiological maturation. Here, we report on the kiwifruit postharvest phase through an integrated approach consisting of pomological analysis combined with NMR/LC-UV/ESI-IT-MS- and 2D-DIGE/nanoLC-ESI-LIT-MS/MS-based proteometabolomic measurements. Kiwifruit samples stored under conventional, cold-based postharvest conditions not involving the use of dedicated chemicals were sampled at four stages (from fruit harvest to pre-commercialization) and analyzed in comparison for pomological features, and outer endocarp metabolite and protein content. About 42 metabolites were quantified, together with corresponding proteomic changes. Proteomics showed that proteins associated with disease/defense, energy, protein destination/storage, cell structure and metabolism functions were affected at precise fruit postharvest times, providing a justification to corresponding pomological/metabolite content characteristics. Bioinformatic analysis of variably represented proteins revealed a central network of interacting species, modulating metabolite level variations during postharvest fruit storage. Kiwifruit allergens were also quantified, demonstrating in some cases their highest levels at the fruit pre-commercialization stage. By lining up kiwifruit postharvest processing to a proteometabolomic depiction, this study integrates previous observations on metabolite and protein content in postharvest berries treated with specific chemical additives, and provides a reference framework for further studies on the optimization of fruit storage before its commercialization.
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http://dx.doi.org/10.3389/fpls.2019.00071DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6369206PMC
February 2019

A proteometabolomic study of Actinidia deliciosa fruit development.

J Proteomics 2018 02 11;172:11-24. Epub 2017 Nov 11.

Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy. Electronic address:

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http://dx.doi.org/10.1016/j.jprot.2017.11.004DOI Listing
February 2018

Carotenoids from the extreme halophilic archaeon Haloterrigena turkmenica: identification and antioxidant activity.

Extremophiles 2017 Sep 12;21(5):933-945. Epub 2017 Aug 12.

Institute of Agro-environmental and Forest Biology, National Research Council of Italy (CNR), Via Pietro Castellino 111, 80131, Naples, Italy.

Haloterrigena turkmenica was able to synthesize carotenoids when grown in halobacteria medium. These molecules have antioxidant properties and find application in food, cosmetic, and pharmaceutical fields. The carotenoids were extracted with methanol, separated by RP-HPLC, and identified by mass spectrometry and UV/Vis spectra analyses. The C carotenoids were the main pigments, and C, C, and C carotenoids were also detected. Seven geometric isomers were distinguished for bacterioruberin, monoanhydrobacterioruberin, and bisanhydrobacterioruberin. The assignment to a specific isomer was tentatively attempted through the analysis of the corresponding UV/Vis spectrum, the intensity of the cis peak, and its spectral fine structure. Lycopene, phytoene, and lycopersene were among the minor carotenoids further identified. The extract displayed antioxidant power higher than alpha-tocopherol, butylhydroxytoluene, and ascorbic acid used as reference compounds. Our studies identified for the first time seven geometric isomers of bacterioruberin derivatives and 30 carotenoids in a haloarchaeon.
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http://dx.doi.org/10.1007/s00792-017-0954-yDOI Listing
September 2017

CK2 and PI3K are direct molecular targets of quercetin in chronic lymphocytic leukaemia.

Oncotarget 2017 Jun;8(26):42571-42587

Institute of Food Sciences, National Research Council, Avellino, Italy.

Despite the encouraging results of the innovative therapeutic treatments, complete remission is uncommon in patients affected by chronic lymphocytic leukaemia, which remains an essentially incurable disease. Recently, clinical trials based on BH3-mimetic drugs showed positive outcomes in subjects with poor prognostic features. However, resistance to treatments occurs in a significant number of patients. We previously reported that the multi-kinase inhibitor quercetin, a natural flavonol, restores sensitivity to ABT-737, a BH3-mimetic compound, in both leukemic cell lines and B-cells isolated from patients. To identify the molecular target of quercetin, we employed a new cell line, HG3, obtained by immortalization of B-cells from a chronic lymphocytic leukaemia patient at the later stage of disease. We confirmed that quercetin in association with ABT-737 synergistically enhances apoptosis in HG3 (combination index < 1 for all fractions affected). We also reported that the cellular uptake of quercetin is extremely rapid, with an intracellular concentration of about 38.5 ng/106 cells, after treatment with 25 μM for 5 min. We demonstrated that the activity of protein kinase CK2, which positively triggers PI3K/Akt pathway by inactivating PTEN phosphatase, is inhibited by quercetin immediately after its addition to HG3 cells (0-2 min). PI3K activity was also inhibited by quercetin within 60 min from the treatment. The combined inhibition of CK2 and PI3K kinase activities by quercetin restored ABT-737 sensitivity and increased lethality in human leukemia cells.
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http://dx.doi.org/10.18632/oncotarget.17246DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5522089PMC
June 2017

A theoretical study on predicted protein targets of apple polyphenols and possible mechanisms of chemoprevention in colorectal cancer.

Sci Rep 2016 09 2;6:32516. Epub 2016 Sep 2.

CNR-ISA, National Research Council, Institute of Food Science, Avellino, Italy.

We investigated the potential role of apple phenolic compounds in human pathologies by integrating chemical characterization of phenolic compounds in three apple varieties, computational approaches to identify potential protein targets of the compounds, bioinformatics analyses on data from public archive of gene expression data, and functional analyses to hypothesize the effects of the selected compounds in molecular pathways. Starting by the analytic characterization of phenolic compounds in three apple varieties, i.e. Annurca, Red Delicious, and Golden Delicious, we used computational approaches to verify by reverse docking the potential protein targets of the identified compounds. Direct docking validation of the potential protein-ligand interactions has generated a short list of human proteins potentially bound by the apple phenolic compounds. By considering the known chemo-preventive role of apple antioxidants' extracts against some human pathologies, we performed a functional analysis by comparison with experimental gene expression data and interaction networks, obtained from public repositories. The results suggest the hypothesis that chemo-preventive effects of apple extracts in human pathologies, in particular for colorectal cancer, may be the interference with the activity of nucleotide metabolism and methylation enzymes, similarly to some classes of anticancer drugs.
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http://dx.doi.org/10.1038/srep32516DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5009435PMC
September 2016

Identification and Quantification of Flavonoids from Two Southern Italian Cultivars of Allium cepa L., Tropea (Red Onion) and Montoro (Copper Onion), and Their Capacity to Protect Human Erythrocytes from Oxidative Stress.

J Agric Food Chem 2015 Jun 22;63(21):5229-38. Epub 2015 May 22.

Institute of Food Sciences, National Research Council, 83100 Avellino, Italy.

Onions (Allium cepa) are consumed worldwide and represent an important source of dietary phytochemicals with proven antioxidant properties, such as phenolic acids, flavonoids, thiosulfinates, and anthocyanins. Epidemiological and experimental data suggest that regular consumption of onions is associated with a reduced risk of degenerative disorders. Therefore, it is of interest to investigate the biological properties of different varieties of onions. Here, we characterized for the first time a variety of onion, called Ramata di Montoro (coppery onion from Montoro), grown in a niche area in southern Italy, and compared its phenolic profile and antioxidant properties to a commercial ecotype of red onion, Tropea, also present in southern Italy. An analytical method based on high-performance liquid chromatography coupled with UV detection and mass spectrometry was used to separate and characterize the phenolic fraction (anthocyanins and flavonols) extracted from both coppery and red types. The main compounds detected in the two ecotypes were quercetin and quercetin glucosides, isorhamnetin glucosides, kaempferol glucoside, and, among anthocyanins, cyanidin glucosides. Tropea ecotype onion showed a higher content of flavonols (632.82 mg/kg fresh weight) than Montoro type onion (252.91 mg/kg fresh weight). Accordingly, the antioxidant activity of the former was 2.8-fold higher compared to the latter. More pronounced were the differences existing between the four anthocyanins detected in the two ecotypes, with those in the Tropea ecotype onion present at concentrations 20-230-fold higher than in the Montoro type onion. Both extracts reduced LDL oxidation about 6-fold and protected human erythrocytes from oxidative damage induced by HClO by about 40%. In addition, as a consequence of HClO treatment, glutathione concentration in erythrocytes was reduced about 50% and pretreatment with onion extracts induced a recovery of glutathione level by about 15-22%. Qualitative differences highlighted in the chemical composition of the two phenolic extracts, especially the total content of anthocyanins, which was 30-fold higher in Montoro type onion compared to Tropea ecotype, can be associated with the protective effects measured against oxidative damage induced in human erythrocytes.
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http://dx.doi.org/10.1021/acs.jafc.5b01206DOI Listing
June 2015

Analysis of different European hazelnut (Corylus avellana L.) cultivars: authentication, phenotypic features, and phenolic profiles.

J Agric Food Chem 2014 Jul 24;62(26):6236-46. Epub 2014 Jun 24.

Unità di Ricerca per la Frutticoltura (Fruit Tree Research Unit), Consiglio per la Ricerca e la Sperimentazione in Agricoltura , Via Torrino, 3, 81100 Caserta, Italy.

Hazelnuts exhibit functional properties due to their content in fatty acids and phenolic compounds that could positively affect human health. The food industry requires precise traits for morphological, chemical, and physical kernel features so that some cultivars could be more suitable for specific industrial processing. In this study, agronomical and morphological features of 29 hazelnut cultivars were evaluated and a detailed structural characterization of kernel polyphenols was performed, confirming the presence of protocatechuic acid, flavan-3-ols such as catechin, procyanidin B2, six procyanidin oligomers, flavonols, and one dihydrochalcone in all the analyzed cultivars. In addition, an innovative methodology based on the MALDI-TOF mass spectrometric analysis of peptide/protein components extracted from kernels was developed for the authentication of the most valuable cultivars. The proposed method is rapid, simple, and reliable and holds the potential to be applied in quality control processes. These results could be useful in hazelnut cultivar evaluation and choice for growers, breeders, and food industry.
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http://dx.doi.org/10.1021/jf5018324DOI Listing
July 2014

Identification of glutathione-methacrylates adducts in gingival fibroblasts and erythrocytes by HPLC-MS and capillary electrophoresis.

Dent Mater 2011 May 2;27(5):e87-98. Epub 2011 Mar 2.

Institute of Biochemistry and Clinical Biochemistry, Catholic University, Rome, Italy.

Objectives: Methacrylic monomers are released, from dental composite resins, either into the oral cavity or in pulpal tissues, where they can cause local or systemic adverse effects. The mechanisms of these effects are not well understood, probably because such molecules can act at different levels also inducing a depletion of intracellular glutathione (GSH). GSH can detoxify methacrylates by conjugating their α,β-unsaturated carbon-carbon moiety to the thiol group, with the catalysis of glutathione S-transferases (GST). This reaction determines a GSH cellular depletion and belongs to the metabolism of α,β-unsaturated esters, protecting the body against the toxic effects of electrophiles. On the basis of the above considerations, this work aim is to set up a method for the detection of the adducts formed by methacrylic monomers with GSH in cells using HPLC coupled to mass spectrometry (HPLC-MS) and micellar electrokinetic capillary chromatography (MECK) techniques.

Methods And Results: Adducts of glutathione with triethylene glycol dimethacrylate (TEGDMA) and hydroxyethyl methacrylate (HEMA) were incontrovertibly identified by HPLC-MS and MECK in human gingival fibroblasts and erythrocytes, both outside and inside cells. Molecular docking simulations of HEMA and TEGDMA in the experimental structure of glutathione S-transferase, are also reported to rationalize the effectiveness of such enzyme in the catalysis of the above described reaction.

Significance: The setup of a method for the identification of GSH-methacrylate adducts allows to determine when the metabolic pathway involving such compounds is employed by cells for the detoxification of monomers leached from composite resins.
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http://dx.doi.org/10.1016/j.dental.2011.01.002DOI Listing
May 2011

Characterization of flavonoid and naphthopyranone derivatives from Eriocaulon ligulatum using liquid chromatography tandem mass spectrometry.

Nat Prod Commun 2009 Dec;4(12):1651-6

São Paulo State University-UNESP, Institute of Chemistry, Organic Chemistry Department, CP 355, CEP 14800-900, Araraquara, SP, Brazil.

Liquid chromatography-electrospray ionization multistage ion trap mass spectrometry (LC-ESI-IT-MS(n)) was used to analyze the secondary metabolites in the methanol extract of the capitulae of Eriocaulon ligulatum. The major components were mono- and diglycosides of flavonoids and naphthopyranones. Eleven compounds, including four new flavonol glycosides, were identified based on their fragmentation patterns in MS experiments and on NMR analysis of the isolated compounds. The described data may contribute to a better understanding of the taxonomic classification of the Eriocaulaceae family.
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December 2009

Oxylipins from Dracontium loretense.

J Nat Prod 2009 May;72(5):813-7

Dipartimento di Scienze Farmaceutiche, Università degli Studi di Salerno, Via Ponte Don Melillo, 84084 Fisciano, SA, Italy.

Four novel oxylipins (1-4) were isolated from the n-butanol extract of the corms of Dracontium loretense. Their structures were assigned by 1D and 2D NMR analyses and electrospray ionization multistage ion trap mass spectrometry (ESI-ITMS(n)) data. Relative configurations were assigned on the basis of combined analysis of homonuclear and heteronuclear (2,3)J couplings, along with ROE data. Oxylipin 2 exhibited an immunostimulatory effect on human PBMC proliferation.
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http://dx.doi.org/10.1021/np8006205DOI Listing
May 2009

Transplastomic tobacco plants expressing a fatty acid desaturase gene exhibit altered fatty acid profiles and improved cold tolerance.

Transgenic Res 2008 Oct 24;17(5):769-82. Epub 2008 Jan 24.

CNR-IGV, Institute of Plant Genetics-Portici, Portici, NA, Italy.

The possibility of altering the unsaturation level of fatty acids in plant lipids by genetic transformation has implications for the stress tolerance of higher plants as well as for their nutritional value and industrial utilisation. While the integration and expression of transgenes in the plastome has several potential advantages over nuclear transformation, very few attempts have been made to manipulate fatty acid biosynthesis using plastid transformation. We produced transplastomic tobacco plants that express a Delta(9) desaturase gene from either the wild potato species Solanum commersonii or the cyanobacterium Anacystis nidulans, using PEG-mediated DNA uptake by protoplasts. Incorporation of chloroplast antibiotic-insensitive point mutations in the transforming DNA was used to select transformants. The presence of the transcript and the Delta(9) desaturase protein in transplastomic plants was confirmed by northern and western blot analyses. In comparison with control plants, transplastomic plants showed altered fatty acid profiles and an increase in their unsaturation level both in leaves and seeds. The two transgenes produced comparable results. The results obtained demonstrate the feasibility of using plastid transformation to engineer lipid metabolic pathways in both vegetative and reproductive tissues and suggest an increase of cold tolerance in transplastomic plants showing altered leaf fatty acid profiles. This is the first example of transplastomic plants expressing an agronomically relevant gene produced with the "binding-type" vectors, which do not contain a heterologous marker gene. In fact, the transplastomic plants expressing the S. commersonii gene contain only plant-derived sequences, a clear attraction from a public acceptability perspective.
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http://dx.doi.org/10.1007/s11248-008-9164-9DOI Listing
October 2008

Novel galactolipids from the leaves of Ipomoea batatas L.: characterization by liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight tandem mass spectrometry.

J Agric Food Chem 2007 Dec 8;55(25):10289-97. Epub 2007 Nov 8.

Dipartimento di Scienze Farmaceutiche, Università degli Studi di Salerno, Via Ponte Don Melillo, 84084 Fisciano, Salerno, Italy.

Sixteen novel and ten known galactolipids have been isolated and characterized from the leaves of Ipomoea batatas L. (sweet potato) using an analytical method based on high-performance liquid chromatography coupled with electrospray ionization-quadrupole time-of-flight tandem mass spectrometry. Using this technique, the structures and regiochemistries of the fatty acyl groups and the positions of the double bonds on the acyl chains were determined. Sugar moieties were identified by analysis of one- and two-dimensional nuclear magnetic resonance spectra. The positions of the double bonds of polyunsaturated fatty acids were confirmed, and in some cases their geometries determined, by gas chromatography-mass spectrometry. This is the first report of galactolipids in the leaves of sweet potato.
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http://dx.doi.org/10.1021/jf071331zDOI Listing
December 2007

Biochemical and structural characterization of mammalian-like purine nucleoside phosphorylase from the Archaeon Pyrococcus furiosus.

FEBS J 2007 May 10;274(10):2482-95. Epub 2007 Apr 10.

Dipartimento di Biochimica e Biofisica F. Cedrangolo, Seconda Università di Napoli, Napoli, Italy.

We report here the characterization of the first mammalian-like purine nucleoside phosphorylase from the hyperthermophilic archaeon Pyrococcus furiosus (PfPNP). The gene PF0853 encoding PfPNP was cloned and expressed in Escherichia coli and the recombinant protein was purified to homogeneity. PfPNP is a homohexamer of 180 kDa which shows a much higher similarity with 5'-deoxy-5'-methylthioadenosine phosphorylase (MTAP) than with purine nucleoside phosphorylase (PNP) family members. Like human PNP, PfPNP shows an absolute specificity for inosine and guanosine. PfPNP shares 50% identity with MTAP from P. furiosus (PfMTAP). The alignment of the protein sequences of PfPNP and PfMTAP indicates that only four residue changes are able to switch the specificity of PfPNP from a 6-oxo to a 6-amino purine nucleoside phosphorylase still maintaining the same overall active site organization. PfPNP is highly thermophilic with an optimum temperature of 120 degrees C and is characterized by extreme thermodynamic stability (T(m), 110 degrees C that increases to 120 degrees C in the presence of 100 mm phosphate), kinetic stability (100% residual activity after 4 h incubation at 100 degrees C), and remarkable SDS-resistance. Limited proteolysis indicated that the only proteolytic cleavage site is localized in the C-terminal region and that the C-terminal peptide is not necessary for the integrity of the active site. By integrating biochemical methodologies with mass spectrometry we assigned three pairs of intrasubunit disulfide bridges that play a role in the stability of the enzyme against thermal inactivation. The characterization of the thermal properties of the C254S/C256S mutant suggests that the CXC motif in the C-terminal region may also account for the extreme enzyme thermostability.
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http://dx.doi.org/10.1111/j.1742-4658.2007.05784.xDOI Listing
May 2007

Liquid chromatography/electrospray ionization tandem mass spectrometry profiling of compounds from the infusion of Byrsonima fagifolia Niedenzu.

Rapid Commun Mass Spectrom 2007 ;21(8):1393-1400

Departamento de Química Orgânica, Instituto de Química de Araraquara, São Paulo State University-UNESP, Rua Francisco Degni s/n, Bairro Quitandinha, CEP 14800-900 Araraquara, SP, Brazil.

A rapid analytical approach suitable to achieve a comprehensive characterization of the compounds present in the infusion prepared from the leaves of Byrsonima fagifolia Niedenzu (Malpighiaceae), a Brazilian plant used as an infusion to treat gastric disorders, was developed. The method was based on high-performance liquid chromatography coupled to electrospray negative ionisation multistage ion trap mass spectrometry (HPLC/ESI-ITMSn). The main ions in the ESI-ITMS spectra were attributed to a quinic acid core containing from one to five galloyl units. Quercetin derivatives containing one and two sugar moieties as well as galloyl esterification were also detected. These results indicated that HPLC/ESI-ITMSn is easily applicable to infusions of this plant and allows the rapid and direct identification of these compounds in crude plant extracts.
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http://dx.doi.org/10.1002/rcm.2971DOI Listing
May 2007

Study of the three-way interaction between Trichoderma atroviride, plant and fungal pathogens by using a proteomic approach.

Curr Genet 2006 Nov 29;50(5):307-21. Epub 2006 Aug 29.

Dipartimento di Arboricoltura, Botanica e Patologia Vegetale, Università degli Studi di Napoli Federico II, Portici (NA), Italy.

The main molecular factors involved in the complex interactions occurring between plants (bean), two different fungal pathogens (Botrytis cinerea, Rhizoctonia solani) and an antagonistic strain of the genus Trichoderma were investigated. Two-dimensional (2-D) electrophoresis was used to analyze separately collected proteomes from each single, two- or three-partner interaction (i.e., plant, pathogenic and antagonistic fungus alone and in all possible combinations). Differential proteins were subjected to mass spectrometry and in silico analysis to search for homologies with known proteins. In the plant proteome, specific pathogenesis-related proteins and other disease-related factors (i.e., potential resistance genes) seem to be associated with the interaction with either one of the two pathogens and/or T. atroviride. This finding is in agreement with the demonstrated ability of Trichoderma spp. to induce systemic resistance against various microbial pathogens. On the other side, many differential proteins obtained from the T. atroviride interaction proteome showed interesting homologies with a fungal hydrophobin, ABC transporters, etc. Virulence factors, like cyclophilins, were up-regulated in the pathogen proteome during the interaction with the plant alone or with the antagonist too. We isolated and confidently identified a large number of protein factors associated to the multi-player interactions examined.
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http://dx.doi.org/10.1007/s00294-006-0091-0DOI Listing
November 2006

Glycated fibroblast growth factor-2 is quickly produced in vitro upon low-millimolar glucose treatment and detected in vivo in diabetic mice.

Mol Endocrinol 2006 Nov 13;20(11):2806-18. Epub 2006 Jul 13.

Dipartimento di Ematologia, Oncologia e Medicina Molecolare, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Roma, Italy.

Angiogenesis impairment in hyperglycemic patients represents a leading cause of severe vascular complications of both type-1 and -2 diabetes mellitus (DM). Angiogenesis dysfunction in DM is related to glycemic control; however, molecular mechanisms involved are still unclear. Fibroblast growth factor-2 (FGF-2) is a potent angiogenic factor and, according to previous evidence, may represent a key target of molecular modifications triggered by high-sugar exposure. Therefore, the purpose of this study was to investigate whether short incubation with hyperglycemic levels of glucose affected FGF-2 and whether glucose-modified FGF-2 was detectable in vivo. Biochemical analyses carried out with SDS-PAGE, fluorescence emission, mass-spectrometry, immunoblot, and competitive ELISA experiments demonstrated that human FGF-2 undergoes a rapid and specific glycation upon 12.5-50 mm glucose exposure. In addition, FGF-2 exposed for 30 min to 12.5 mm glucose lost mitogenic and chemotactic activity in a time- and dose-dependent manner. Under similar conditions, binding affinity to FGF receptor 1 was dramatically reduced by 20-fold, as well as FGF receptor 1 and ERK-1/2 phosphorylation, and FGF-2 lost about 45% of angiogenic activity in two different in vivo angiogenic (Matrigel and chorioallantoic-membrane) assays. Such glucose-induced modification was specific, because other angiogenic growth factors, namely platelet-derived growth factor BB and placental-derived growth factor were not significantly or markedly less modified. Finally, for the first time, glycated-FGF-2 was detected in vivo, in tissues from hyperglycemic nonobese diabetic mice, in significantly higher amounts than in normoglycemic mice. In conclusion, hyperglycemic levels of glucose may strongly affect FGF-2 structure and impair its angiogenic features, and endogenous glycated-FGF-2 is present in diabetic mice, indicating a novel pathogenetic mechanism underlying angiogenesis defects in DM.
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http://dx.doi.org/10.1210/me.2005-0322DOI Listing
November 2006

Small surface-associated factors mediate adhesion of a food-isolated strain of Lactobacillus fermentum to Caco-2 cells.

Res Microbiol 2005 Aug;156(7):830-6

Dipartimento di Biologia Strutturale e Funzionale, Università Federico II, Napoli, Italy.

In a search for bacteria having putative probiotic activity, we screened a collection of food-isolated microorganisms for the ability to survive at low pH in the presence of bile salts and for the production of antimicrobial compounds active against a number of animal pathogens. Among these, we found a strain that we classified as a member of Lactobacillus fermentum sp., and we further investigated its features. This organism was able to adhere to human enterocyte-like (Caco-2) cells with high efficiency as compared to that of a well known indicator strain. Chromatographic analysis indicated that at least two small (less than 3 kDa) factors were involved in mediating the in vitro interaction of L. fermentum with Caco-2 cells. Adhesion activity could be abolished by mild treatment of the bacterial cells in buffer and rescued by incubating them with either the same buffer after its use in the treatment or with chromatographic fractions containing each of the two factors, which indicated that these factors were loosely associated with the cell wall and that each of them was sufficient to warrant the adhesiveness of L. fermentum to Caco-2 cells. These data are suggestive of a novel mechanism of bacterial adhesion to epithelial cells.
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http://dx.doi.org/10.1016/j.resmic.2005.05.001DOI Listing
August 2005

Flavonoids from the leaves of Cyclanthera pedata: two new malonyl derivatives.

Phytochem Anal 2005 May-Jun;16(3):210-6

Dipartimento di Scienze Farmaceutiche, Università di Salerno, Via Ponte don Melillo, 84084 Fisciano (SA), Italy.

Reversed-phase HPLC coupled with electrospray MS has been used for the simultaneous separation and determination of flavonoid metabolites in leaves of Cyclanthera pedata, an edible Peruvian plant mainly used in South America for its anti-inflammatory, hypoglycaemic and hypocholesterolaemic properties. The flavonoid content of the leaves of C. pedata was compared qualitatively and quantitatively with that of the fruits. The isolation and structural characterisation by MS and NMR of two new minor components of the fruits, namely, 6-C-fucopyranosyl-(3-malonyl)-chrysin and 6-C-fucopyranosyl-(4-malonyl)-chrysin, are described.
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http://dx.doi.org/10.1002/pca.847DOI Listing
August 2005

Hb Cardarelli [beta86(F2)Ala-->Pro]: a new unstable and hyperaffine variant in association with beta(+)-thalassemia.

Hemoglobin 2004 May;28(2):103-15

Centro Microcitemie A. Mastrobuoni, Azienda Ospedale Cardarelli, Napoli, Italia.

Hb Cardarelli [beta86(F2)Ala-->Pro] is a new unstable and high oxygen affinity variant found in several members of a family from Naples, Southern Italy. A detailed structural and functional characterization of the variant was performed on two subjects, at both the protein and DNA level. The first patient exhibited 43% of the variant hemoglobin (Hb) without major hematological problems. The proband showed 82% of the abnormal Hb in association with beta(+)-thalassemia (thal) that caused relevant erythrocytosis requiring frequent phlebotomies. Structural investigation of the Hb variant by mass spectrometric methodologies identified the amino acid replacement as Ala-->Pro at beta86. The corresponding DNA mutation GCC-->CCC at codon 86 of the beta-globin gene was assessed by both DNA sequencing and amplification refractory mutation system (ARMS) techniques. Functional studies carried out on whole blood and diluted hemolysates from both patients demonstrated increased oxygen affinity, decreased Bohr effect, reduced heme-heme interaction and nearly halved 2,3-diphosphoglycerate (2,3-DPG) and chloride effects.
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http://dx.doi.org/10.1081/hem-120035915DOI Listing
May 2004

Analysis of flavonoids from Cyclanthera pedata fruits by liquid chromatography/electrospray mass spectrometry.

J Pharm Biomed Anal 2004 Feb;34(2):295-304

Centro di Spettrometria di Massa Proteomica e Biomolecolare, Istituto di Scienze dell'alimentazione-C.N.R., Via Roma 52a-c, 83100 Avellino, Italy.

A liquid chromatography/mass spectrometry (LC/MS) based method was developed for the characterization of fruits of Cyclanthera pedata Scrabs (Caigua), a Peruvian food and medicinal plant. This method is based on the separation of flavonoid glycosides present in the methanolic extracts from C. pedata fruits using high performance liquid chromatography (HPLC) followed by detection with electrospray ionization mass spectrometry (ESI/MS). Chromatographic separation of the analytes of interest was achieved on a Symmetry C-18 column with detection in positive ion mode. Calibration graphs were obtained by determining the area ratio between external standard of each major compound and the internal standard naringine. Due to the sensitivity and the repeatability of the assay, this method is suitable for industrial quality control of raw materials and final products.
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http://dx.doi.org/10.1016/S0731-7085(03)00580-6DOI Listing
February 2004

Investigation of the tuber constituents of maca (Lepidium meyenii Walp.).

J Agric Food Chem 2002 Sep;50(20):5621-5

Dipartimento di Scienze Farmaceutiche, Università degli Studi di Salerno, via Ponte Don Melillo, 84084 Fisciano, Italy.

Lepidium meyenii, known in South America as maca, has received attention worldwide as a powerful energizer that improves physical and mental conditions and increases fertility. Because of these reports, we investigated the secondary metabolites of the tuber of maca. The methanol extract of the tuber of maca contained, in addition to free sugars and amino acids, the following: uridine, malic acid and its benzoyl derivative, and the glucosinolates, glucotropaeolin and m-methoxyglucotropaeolin. Because glucosinolates and their derived products have received increasing attention due to their biological activities, the occurrence of glucosinolate degradation products in the hexane extract was also investigated, and benzylisothiocyanate and its m-methoxy derivative were isolated. The two glucosinolates were semiquantified by HPLC, and benzylisothiocyanate was semiquantified by GC/MS. The methanol extract of maca tuber also contained (1R,3S)-1-methyltetrahydro-beta-carboline-3-carboxylic acid, a molecule which is reported to exert many activities on the central nervous system.
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http://dx.doi.org/10.1021/jf020280xDOI Listing
September 2002

Triterpenoid saponins from the shells of Argania spinosa seeds.

J Agric Food Chem 2002 Jul;50(16):4600-3

Laboratoire de Chimie des Plantes et de Synthèse Organique et Bioorganique, Faculté des Sciences, University of Rabat, B.P. 1041, RP Rabat, Morocco.

Two new oleanene saponins were isolated from the MeOH extract of the shell of Argania spinosa. They possess protobassic acid and 16alpha-protobassic acid as aglycons. The disaccharide moiety linked to C-3 of the aglycon is made up of two glucose units; the pentasaccharide moiety linked to C-28 is made up of arabinose, xylose, and three rhamnose units. Their structures were elucidated by 1D and 2D NMR experiments including (1)H-(1)H (DQF-COSY, 1D TOCSY, and 2D HOHAHA) and (1)H-(13)C (HSQC and HMBC) spectroscopy along with mass spectrometry.
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http://dx.doi.org/10.1021/jf0200117DOI Listing
July 2002

Hb Vila Real [beta36(C2)Pro-->His] in Italy: characterization of the amino acid substitution and the DNA mutation.

Hemoglobin 2002 Feb;26(1):21-31

Dipartimento di Chimica Organica e Biochimica, Università di Napoli Federico II, Napoli, Italia.

A rare high oxygen affinity hemoglobin variant was identified in a 22-year-old male patient from Napoli (Naples, Italy) affected by erythrocytosis. A detailed structural characterization of the variant hemoglobin was carried out, both at the protein and DNA levels essentially by mass spectrometric procedures and allele-specific amplification techniques. The amino acid substitution was determined by liquid chromatography tandem mass spectrometric analysis of the tryptic digest as beta36(C2)Pro-->His; the corresponding DNA mutation was identified as C-->A at the second position of codon 36 of the beta chain (CCT-->CAT). These variations identified the presence of Hb Vila Real, described only once before in a Portuguese woman. Haplotype analysis of DNA polymorphisms showed that the beta-globin gene of Hb Vila Real was associated with haplotype I.
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http://dx.doi.org/10.1081/hem-120002937DOI Listing
February 2002
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