Macrolinea Cod.: 076.ISA.03

Autori: Bandiera A, Tell G, Marsich E, Scaloni A, Pocsfalvi G, Akintunde Akindahunsi A, Cesaratto L, Manzini G.

Autori CNR:  GABRIELLA KATALIN POCSFALVI

Titolo: Cytosine-block telomeric type DNA-binding activity of hnRNP proteins from human cell lines

Abstract:

Following the observation of the presence in mammalian nuclear extracts of a DNA binding activity quite specific for the single-stranded C-rich telomeric motif, we have isolated from the K562 human cell line by affinity chromatography and identified by mass spectrometry a number of proteins able to bind to this sequence. All of them belong to different heterogeneous nuclear ribonucleoprotein subgroups (hnRNP). Whereas many of them, namely hnRNP K, two isoforms of hnRNP I, and the factor JKTBP, appear to bind to this sequence with limited specificity after isolation, an isoform of hnRNP D (alias AUF1) and particularly hnRNP E1 (alias PCBP-1) show a remarkable specificity for the (CCCTAA)n repeated motif. Both have been obtained also as recombinant proteins expressed in Escherichia coli and have been shown to retain their binding specificity toward the C-block repeated sequence. In the light of the current knowledge about these proteins, their possible involvement in telomere functioning is discussed

Rivista: ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS

anno: 2003

volume n.: 409

pag. da: 305

a:314

Macrolinea Cod.: 076.ISA.04

Autori: Caira, S., Ferranti, P., Gatti, M., Fornasari, ME, Barone, F., Lilla, S., Mucchetti, G., Picariello, G., Chianese, L., Neviani, E., Addeo, F.

Autori CNR:  GIANLUCA PICARIELLO;  SIMONETTA CAIRA; 

Titolo: Synthetic peptides as substrate for assaying the proteolytic activity of Lactobacillus helveticus

Abstract:

Four Lactobacillus helveticus strains were studied for proteolytic capacity and general aminopeptidase (AP) and X-Pro dipeptidyl aminopeptidase (DAP) activity. The rate of hydrolysis and the activity against synthetic substrates with N-terminal residues of Arg, Lys, Leu, Glu or Pro, varied markedly among the strains. The X-Pro DAP activity was consistently high. The crude cell-wall and cytoplasm extracts from strain Lb. helveticus ISLC59 were analysed thoroughly for their proteolysis ability by using four synthetic peptide substrates, including alpha(s)1-CN(f1-23). Peptides formed during in vitro hydrolysis of the synthetic substrates by cell wall and cytoplasm preparations were identified by LC-ESI/MS. In doing so, it was possible to infer a prevalent endopeptidase activity splitting Lys7-His8 and Gln13-Glu14 bonds in the cytoplasm, and to deduce a secondary activity, which hydrolysed Glu14-Val15, Leu16-Asn17, Glu18-Asn19 and Lys3-His4 bonds lacking in the cell-wall. The presence of exopeptidases, as mainly AP, DAP, and carboxypeptidase (CPase) was deduced from the formation of several N- and C-terminally truncated peptides sets. The AP activity was higher in the cell-wall layer, where CPase activity was absent. The in vitro assays with cell extracts of the Lb. helveticus ISLC59 strain revealed extensive exopeptidase and endopeptidase activities. In several cases, the hydrolytic system of Lb. helveticus that splits in vitro alpha(s)1-CN(f1-23) peptide bonds was similar to that of Lactococcus lactis. The effects were also compared with those occurring in vivo in hard cheese such as Grana Padano.

Rivista: JOURNAL OF DAIRY RESEARCH

anno: 2003

volume n.: 70

pag. da: 315

a:325

Macrolinea Cod.: 076.ISA.03

Autori: Caporale C, Facchiano A, Leonardi L, Bertini L, Chilosi G, Buonocore V, Caruso C.

Autori CNR:  ANGELO FACCHIANO; 

Titolo: Comparing the modelled structures of PR-4 proteins from wheat

Abstract:

We have constructed three-dimensional models of four pathogenesis-related (PR) proteins from wheat (wheatwins) belonging to the PR-4 family. All the models were based on the knowledge of the tertiary structure of barwin, a highly homologous protein from barley. Wheatwin1 and wheatwin2 differ in two amino acid residues (positions 62 and 68) out of 125. Wheatwin4 differs from wheatwin2 in one residue at position 78, while wheatwin3 differs from wheatwin1 in one residue at position 88. The global folding and the secondary structures were very similar through all the sequences, including the regions of the amino acid substitutions. The main differences were found in the traits 15-21, 84-86 and 91-93. Trait 15-21 was predicted as ss-sheet in wheatwin4 and random-coil in the other proteins. Trait 84-86 was predicted as ss-sheet in wheatwin3 and random-coil in the other proteins. Trait 91-93 was predicted as random coil in wheatwin1 and wheatwin3 and ss-sheet in the other two proteins. Traits 15-21 and 84-86 were exposed, while trait 91-93 was quite hidden in all the proteins. The antifungal activities of the four proteins towards the specific pathogenic fungus Fusarium culmorum were distinct and well correlated to the structural differences. These results suggest that these regions may have a role in the action

Rivista: JOURNAL OF MOLECULAR MODELING

anno: 2003

volume n.: 9

pag. da: 9

a:15

Macrolinea Cod.: 076.ISA.03

Autori: Carla Esposito, Ph.D., Francesco Paparo, Ivana Caputo, Ph.D., Raffaele Porta, Ph.D.,

Autori CNR:  GIUSEPPE MAZZARELLA; 

Titolo: Expression and Enzymatic Activity of Small Intestinal Tissue Transglutaminase in Celiac Disease

Abstract:

OBJECTIVES: The molecular and functional properties of small intestinal tissue transglutaminase are largely unknown despite growing interest because of its role in celiac disease (CD). In this study, we aimed to evaluate tissue transglutaminase expression and enzymatic activity in bioptic fragments obtained from the duodenum of untreated individuals with CD and from control subjects. METHODS: Analysis of tissue transglutaminase mRNA expression was performed by reverse transcription-polymerase chain reaction (RT-PCR). The presence of the enzyme in bioptic fragments as well as in homogenates from CD patients and controls was revealed by immunohistochemistry and Western blot, respectively, using the antitissue transglutaminase CUB 7402 clone. To evaluate in situ transglutaminase activity, sections of bioptic fragments were incubated in the presence of 5 mmol/L CaCl(2) with 5-(biotinamido)pentylamine or, alternatively, with a biotinylated glutamine-containing hexapeptide (TVQQEL) and the biotinylated 31-43 A-gliadin-derived peptide. RESULTS: Tissue transglutaminase mRNA levels were 1.0-fold higher (p < 0.05) in CD patients than in controls. Immunohistochemistry and in situ demonstration of enzymatic activity in celiac mucosa clearly showed an increased expression of active tissue transglutaminase in the extracellular matrix of the subepithelial region and in the enterocytes. Staining of the biotinylated 31-43 A-gliadin peptide in the same area of tissue transglutaminase suggested the presence of lysine-donor substrates in intestinal mucosa. CONCLUSIONS: Tissue transglutaminase is more expressed and active in defined areas of the small intestinal mucosa from patients with CD. The presence in the celiac mucosa of proteins able to act as amine-donor substrates suggests that tissue transglutaminase-mediated post-translational modification of proteins cross-linked with gliadin peptides may represent a pathogenic mechanism of CD.

Rivista: AMERICAN JOURNAL OF GASTROENTEROLOGY

anno: 2003

volume n.: 98

pag. da: 1813

a:1820

Macrolinea Cod.: 076.ISA.04

Autori: Cereda, A., Forlani, F., Iametti, S., Bernhardt, R., Ferranti, P., Picariello, G., Pagani, S., Bonomi, F.

Autori CNR:  GIANLUCA PICARIELLO; 

Titolo: Molecular recognition between Azotobacter vinelandii rhodanese and a sulfur acceptor protein

Abstract:

The occurrence of rhodanese-like proteins in the major evolutionary phyla, together with the observed abundance of these proteins also within the same genome, suggests that their function cannot be limited to cyanide scavenging. The aim of the present study was to investigate whether Azotobacter vinelandii RhdA, an enzyme possessing unique biochemical and structural features with respect to other members of rhodanese homology superfamily, could recognize a suitable protein as a potential acceptor of the sulfane sulfur held on its catalytic Cys residue. Both the potential sulfur-delivery RhdA-S and the sulfur-deprived RhdA were found to interact with either holo- or apo-adrenodoxin, the 'substrate' protein used in this work. Interaction of RhdA-S with apo-adrenodoxin led to mobilization of RhdA-S sulfane sulfur. Under appropriate conditions, the sulfur released from RhdA-S was productively used for 2Fe-2S cluster reconstitution to yield holo-adrenodoxin from apo-adrenodoxin in the absence of any other sulfur source. A comparison of the reactivity of RhdA-S with protein and non-protein thiols allowed also some insights into the accessibility of the sulfane sulfur carried by RhdA.

Rivista: BIOLOGICAL CHEMISTRY

anno: 2003

volume n.: 384

pag. da: 1473

a:1481

Macrolinea Cod.: 076.ISA.01

Autori: Clemente G, Mancini M, Nazzaro F, Lasorella G, Rivieccio A, Palumbo AM, Rivellese AA, Ferrara L, Giacco R.

Autori CNR:  GENNARO CLEMENTE;  ROSALBA GIACCO;  FILOMENA NAZZARO; 

Titolo: Effects of different dairy products on postprandial lipemia

Abstract:

BACKGROUND AND AIM: To evaluate the effects on postprandial lipemia (PPL), of three fat rich meals, with similar composition but different physical structure (liquid, semisolid and solid). METHODS AND RESULTS: Eight type 2 diabetic patients of both genders (6M/2F), age 51+/-9 yrs (M+/-SD), BMI 29+/-3 kg/m2, with fasting plasma glucose levels 145+/-24 mg/dL, cholesterol 200+/-38 mg/dL and triglyceride 110+/-45 mg/dL. Participants consumed in the morning, after a 12-hour fast and at 1-week intervals, three test meals with similar volume and composition [protein 36 g, lipid 30 g, carbohydrate 115 g, energy 3556 kJ (850 Kcal)] but with the main source of fat represented by foods with different physical structure (milk, mozzarella-cheese, butter). Each patient underwent gastric emptying measurements by echography; plasma FFA, triglycerides, glucose and insulin were evaluated at baseline and every hour for six hours after each meal. Fasting plasma glucose, cholesterol and triglyceride concentrations were similar at the baseline of the three test meals. Average increases in postprandial plasma triglyceride levels after butter (88+/-8 mg/dL) and mozzarella-cheese (104+/-56 mg/dL) were not different than after milk (98+/-53 mg/dL). The plasma triglyceride peak was also similar after the three test meals but peak time after butter (315+/-42 min; p<0.01) and mozzarella-cheese (277+/-31 min; p<0.02) was significantly delayed compared to milk (225+/-28 min). Gastric emptying rate was similar after butter and milk (14+/-2, 13+/-6 mL/h) and significantly faster after mozzarella-cheese (18+/-5 mL/h; p<0.03). CONCLUSIONS: While the physical structure of fat-rich foods has no major effect on postprandial plasma triglyceride concentrations, it is able to influence the timing of triglyceride peak; gastric emptying time does not play a major role in modulating the postprandial response of triglycerides and glucose.

Rivista: NUTRITION METABOLISM AND CARDIOVASCULAR DISEASES

anno: 2003

volume n.: 13(6)

pag. da: 377

a:383

Macrolinea Cod.: 076.ISA.04

Autori: Dettin, M., Ferranti, P., Scarinci, C., Picariello, G., Di Bello, C.

Autori CNR:  GIANLUCA PICARIELLO; 

Titolo: Is the V3 loop involved in HIV binding to CD4?

Abstract:

The entry of the human immunodeficiency virus into cells requires the interaction of the viral envelope glycoprotein gp120 with CD4 and a chemokine receptor. The gp120 binding site has been previously mapped to the Ig-CDR2-like region of CD4 first domain. A second area of this domain (Ig-CDR3-like region) is involved in gp120-CD4 interactions, but its gp120 counterpart remained so far unknown. Using a photoaffinity labeling experiment, we demonstrate that a peptide, mapping the (307-330)m region of HIV-MN-gp120 V3 loop, binds a sequence including a part of the Ig-CDR3-like region. These results may contribute to explain the complex mechanism of human immunodeficiency virus penetration, helping the development of new therapeutic agents.

Rivista: BIOCHEMISTRY

anno: 2003

volume n.: 42

pag. da: 9007

a:9012

Macrolinea Cod.: 076.ISA.03

Autori: Di Biase AM, Pietrantoni A, Tinari A, Siciliano R, Valenti P, Antonini G, Seganti L, Superti F.

Autori CNR:  ROSA ANNA SICILIANO; 

Titolo: Heparin-interacting sites of bovine lactoferrin are involved in anti-adenovirus activity.

Abstract:

Lactoferrin, a member of the transferrin family of approximately 80 kDa, consists of a single polypeptide chain folded in two symmetric, globular lobes (N- and C-lobes), each able to bind one ferric ion. This glycoprotein, found in physiological fluids of mammals, plays an important role in immune regulation and in defense mechanisms against bacteria, fungi, parasites, and viruses. Although the antiviral activity of lactoferrin is one of the major biological functions of such protein, the mechanism of action is still under debate. We have investigated both the role of tryptic fragments of bovine lactoferrin and the mechanism of lactoferrin antiviral effect toward adenovirus infection in HEp-2 cells. The results obtained demonstrated that the anti-adenovirus activity of lactoferrin is mediated by the N-terminal half of the protein as the N-lobe was able to inhibit adenovirus infection, even if at lower extent than undigested lactoferrin, whereas C-lobe was ineffective. The results also showed that the anti-adenovirus action of lactoferrin and of its N-terminal peptide lactoferricin took place on virus attachment to cell membrane, mainly through competition for common glycosaminoglycan receptors. The data provide evidence that the anti-adenovirus activity of lactoferrin is mediated mainly by the cluster of positive charges at the N-terminus of whole molecule and that the N-terminal peptide lactoferricin alone is sufficient to prevent infection.

Rivista: JOURNAL OF MEDICAL VIROLOGY

anno: 2003

volume n.: 69(4)

pag. da: 495

a:502

Macrolinea Cod.: 076.ISA.03

Autori: Facchiano A, Facchiano A, Facchiano F.

Autori CNR:  ANGELO FACCHIANO; 

Titolo: (Active Sequences Collection) database: a new tool to assign functions to protein sequences.

Abstract:

Active Sequences Collection (ASC) is a collection of amino acid sequences, with an unique feature: only short sequences are collected, with a demonstrated biological activity. The current version of ASC consists of three sections: DORRS, a collection of active RGD-containing peptides; TRANSIT, a collection of protein regions active as substrates of transglutaminase enzyme (TGase), and BAC, a collection of short peptides with demonstrated biological activity. Literature references for each entry are reported, as well as cross references to other databases, when available. The current version of ASC includes more than 800 different entries. The main scope of this collection is to offer a new tool to investigate the structural features of protein active sites, additionally to similarity searches against large protein databases or searching for known functional patterns. ASC database is available at the web address http://crisceb.unina2.it/ASC/ which also offers a dedicated query interface to compare user-defined protein sequences with the database, as well as an updating interface to allow contribution of new referenced active sequences.

Rivista: NUCLEIC ACIDS RESEARCH

anno: 2003

volume n.: 31

pag. da: 379

a:382

Note:

Copyright: la banca dati oggetto della pubblicazione e' stata depositata secondo le procedure in uso in Italia

Macrolinea Cod.: 076.ISA.03

Autori: Facchiano A, Russo K, Facchiano AM, De Marchis F, Facchiano F, Ribatti D, Aguzzi MS, Capogrossi MC

Autori CNR: ANGELO FACCHIANO; 

Titolo: Identification of a novel domain of fibroblast growth factor- 2 controlling its angiogenic properties.

Abstract:

Fibroblast growth factor 2 (FGF-2) is a potent factor modulating the activity of many cell types. Its dimerization and binding to high affinity receptors are considered to be necessary steps to induce FGF receptor phosphorylation and signaling activation. A structural analysis was carried out and a region encompassing residues 48-58 of human FGF-2 was identified, as potentially involved in FGF-2 dimerization. A peptide (FREG-48-58) derived from this region strongly and specifically inhibited FGF-2 induced proliferation and migration of primary bovine aorta endothelial cells (BAEC) in vitro, and markedly reduced FGF-2-dependent angiogenesis in two distinct in vivo assays. To further investigate the role of region 48-58, a polyclonal antibody raised against FREG-(48-58) was tested and was found to block FGF-2 action in vitro. Human FGF-2 has three histidine residues, one falling within the region 48-58. Chemical modification of histidine residues blocked FGF-2 activity and FREG-(48-58) inhibitory effect in vitro, indicating that histidine residues, in particular the one within FREG-(48-58) region, play a crucial role in the observed activity. Additional experiments showed that FREG-(48-58) specifically interacted with FGF-2, impaired FGF-2-interaction with itself, with heparin and with FGF receptor 1, and inhibited FGF-2-induced receptor phosphorylation and FGF-2 internalization. These data indicate for the first time that region 48-58 of FGF-2 is a functional domain controlling FGF-2 activity.

Rivista: JOURNAL OF BIOLOGICAL CHEMISTRY

anno: 2003

volume n.: 278

pag. da: 8751

a:8760

Macrolinea Cod.: 076.ISA.04

Autori: Mamone, G., Caira, S., Garro, G., Nicolai, A., Ferranti, P., Picariello, G., Malorni, A., Chianese, L., Addeo, F.

Autori CNR:  GIANLUCA PICARIELLO;  GIANFRANCO MAMONE;  SIMONETTA CAIRA;  ANTONIO MALORNI; 

Titolo: Casein phosphoproteome: identification of phosphoproteins by combined mass spectrometry and two-dimensional gel electrophoresis

Abstract:

We report a fast and easy-to-use procedure that combines polyacrylamide gel electrophoresis with matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF) and nanoelectrospray-tandem mass spectrometry (nES-MS/MS) analysis for the identification of casein components and defined phosphorylated sites. This methodology ensured identification of more than 30 phosphorylated proteins, five beta-, fifteen alpha(s1)-, ten alpha(s2)-, and four kappa-casein (CN) components, including nonallelic, differently phosphorylated, and glycosylated forms. The sugar motif covalently bound to kappa-CN was identified as chains, trisaccharide GalNAc, Gal, NeuGc, and tetrasaccharide 1GalNAc, 1Gal, 2NeuGc. Also identified was a biantennary chain made up of both chains of trisaccharide 1GalNAc, 1Gal, 1NeuGc, and tetrasaccharide 1GalNAc, 1Gal, 2NeuGc moiety on a single kappa-CN component. The phosphate group on site Ser12 of tryptic peptide 8-22 of most phosphorylated alpha(s1)-CN (11 phosphate groups) was localized and the oligosaccharide sequence of the main tryptic glycopeptides of two kappa-CN components was determined by means of MS/MS analysis.

Rivista: ELECTROPHORESIS

anno: 2003

volume n.: 24

pag. da: 2824

a:2837

Macrolinea Cod.: 076.ISA.03

Autori: Mazzarella G, MacDonald TT, Salvati VM, Mulligan P, Pasquale L, Stefanile R, Lionetti P, Auricchio S, Pallone F, Troncone R, Monteleone G.

Autori CNR:  GIUSEPPE MAZZARELLA; 

Titolo: Constitutive activation of the signal transducer and activator of transcription pathway in celiac disease lesions.

Abstract:

The biological effects of interferon (IFN)-gamma rely mainly on the activity of the transcription factor signal transducer and activator of transcription (STAT) 1 and the intracellular levels of suppressor of cytokine signaling (SOCS)-1, a negative regulator that controls the amplitude and duration of STAT-1 activation. IFN-gamma is a key mediator of the immunopathology in celiac disease (CD, gluten-sensitive enteropathy). Thus we have investigated STAT-1 signaling and SOCS-1 expression in this condition. As expected, high local concentrations of IFN-gamma were invariably seen in duodenal biopsies from CD patients in comparison to controls. On the basis of immunohistochemistry, STAT-1 phosphorylation, nuclear localization, and DNA-binding activity, STAT-1 activation was consistently more pronounced in CD compared with controls. Despite samples from CD patients containing abundant SOCS-1 mRNA, SOCS-1 protein was expressed at the same level in CD patients and controls. In explant cultures of CD biopsies, gliadin induced the activation of STAT-1 but not SOCS-1. Furthermore, inhibition of STAT-1 prevented the gliadin-mediated induction of ICAM-1 and B7-2. These data suggest that persistent STAT-1 activation can contribute to maintaining and expanding the local inflammatory response in CD.

Rivista: AMERICAN JOURNAL OF PATHOLOGY

anno: 2003

volume n.: 162-6

pag. da: 1845

a:1855

Macrolinea Cod.: 076.ISA.03

Autori: Mazzarella G, Maglio M, Paparo F, Nardone G, Stefanile R, Greco L, van de Wal Y, Kooy Y, Koning F, Auricchio S, Troncone R.

Autori CNR:  GIUSEPPE MAZZARELLA; 

Titolo: An immunodominant DQ8 restricted gliadin peptide activates small intestinal immune response in in vitro cultured mucosa from HLA-DQ8 positive but not HLA-DQ8 negative coeliac patients.

Abstract:

BACKGROUND: Studies on intestinal T cell clones from the mucosa of patients with coeliac disease have led to the identification of immunogenic gliadin epitopes. One is HLA-DQ8 restricted, its recognition by T cells being increased by introduction of negatively charged residues operated by tissue transglutaminase. AIM: To test HLA-DQ8 restricted epitope in both native (QYPSGQGSFQPSQQNPQA) and deamidated (QYPSGEGSFQPSQENPQA) forms in an organ culture system of treated coeliac mucosa from HLA-DQ8 positive and HLA-DQ8 negative patients. PATIENTS AND METHODS: Jejunal biopsies obtained from 10 patients with coeliac disease (six HLA-DQ8 positive and four HLA-DQ8 negative) were cultured in vitro with a peptic-tryptic digest (PT) of gliadin, or with the native (peptide A) or deamidated (peptide B) peptide. Intraepithelial CD3(+) and lamina propria total CD25(+) and CD3(+)CD25(+) cells were counted, lamina propria intercellular adhesion molecule 1 (ICAM-1) expression was evaluated, as well as that of Fas molecules on epithelial cells. RESULTS: In HLA-DQ8 positive, but not in HLA-DQ8 negative, coeliacs the density of intraepithelial CD3(+) cells, lamina propria total CD25(+), and CD3(+)CD25(+) cells, as well as expression of ICAM-1 and Fas molecules were significantly increased in biopsies cultured with PT, peptide A, or peptide B compared with biopsies cultured in medium alone. CONCLUSION: These data show that the DQ8 restricted gliadin peptide is immunogenic only in the intestinal mucosa of HLA-DQ8 positive coeliac patients in both native and deamidated forms.

Rivista: GUT

anno: 2003

volume n.: 52(1)

pag. da: 57

a:62

Note:

cover illustration

Macrolinea Cod.: 076.ISA.03

Autori: Mazzeo M.F., De Giulio B., Senger S., Rossi M., Malorni A., Siciliano R.A.

Autori CNR:  ROSA ANNA SICILIANO;  BEATRICE DE GIULIO;  ANTONIO MALORNI;  MAURO ROSSI; 

Titolo: Identification of transglutaminase-mediated deamidation sites in a recombinant alpha-gliadin by advanced mass-spectrometric methodologies.

Abstract:

Celiac disease is a permanent immune-mediated food intolerance triggered by ingestion of wheat gliadins in genetically susceptible individuals. It has been reported that tissue transglutaminase plays an important role in the onset of celiac disease by converting specific glutamine residues within gliadin fragments into glutamic acid residues. This process increases binding affinity of gliadin peptides to HLA-DQ2/DQ8 molecules, thus enhancing the immune response. The aim of the present study was to achieve a detailed structural characterization of modifications induced by transglutaminase on gliadin peptides. Therefore, structural analyses were carried out on a recombinant alpha-gliadin and on a panel of 26 synthetic peptides, overlapping the complete protein sequence. Modified glutamine residues were identified by means of advanced mass-spectrometric methodologies on the basis of MALDI-TOF-MS and tandem mass spectrometry. Results led to the identification of 19 of 94 glutamine residues present in the recombinant alpha-gliadin, which were converted into glutamic acid residues by a transglutaminase-mediated reaction. This allowed us to achieve a global view of the modifications induced by the enzyme on this protein. Furthermore, results gathered could likely be utilized as relevant information for a better understanding of processes leading to T-cell recognition of gliadin peptides involved in celiac disease.

Rivista: PROTEIN SCIENCE

anno: 2003

volume n.: 12

pag. da: 2434

a:2442

Macrolinea Cod.: 076.ISA.03

Autori: Opsenica D, Angelovski G, Pocsfalvi G, Juranic Z, Zizak Z, Kyle D, Milhous WK, Solaja BA.

Autori CNR:  GABRIELLA KATALIN POCSFALVI; 

Titolo: Antimalarial and antiproliferative evaluation of bis-steroidal tetraoxanes

Abstract:

Several cis and trans bis-steroidal 1,2,4,5-tetraoxanes possessing amide terminus were synthesised and evaluated as antimalarials and antiproliferatives. The compounds exhibited submicromolar antimalarial activity against Plasmodium falciparum D6 and W2 strains. The existence of HN-C(O) moiety was found necessary for pronounced antimalarial and antiproliferative activity. In antiproliferative screen, the trans tetraoxane 6 was found to exhibit a pronounced cytotoxicity on 14 cancer cell lines. In addition, tetraoxanes 11 and 12 exhibited significant cytotoxic activity too; microscopic examination of treated HeLa cells showed morphological appearance reminiscent for apoptosis (condensed and/or fragmented nuclei).

Rivista: (vedi all.1) 01811

Rivista: BIOORGANIC & MEDICINAL CHEMISTRY

anno: 2003

volume n.: 11

pag. da: 2761

a:2768

Macrolinea Cod.: 076.ISA.03

Autori: Salvati VM, MacDonald TT, del Vecchio Blanco G, Mazzarella G, Monteleone I, Vavassori P, Auricchio S, Pallone F, Troncone R, Monteleone G.

Autori CNR:  GIUSEPPE MAZZARELLA; 

Titolo: Enhanced expression of interferon regulatory factor-1 in the mucosa of children with celiac disease.

Abstract:

Celiac disease (CD) is an enteropathy characterized by a Th1-type immune response to the dietary gluten. The transcriptional mechanisms or factors that control Th1 cell development in this condition remain to be elucidated. The aim of this study was to analyze in CD the expression of interferon (IFN) regulatory factor (IRF)-1, a transcription factor that regulates the differentiation and function of Th1 cells. Duodenal biopsies were taken from children with untreated CD and control children, and analyzed for IRF-1 by Southern blotting of reverse-transcriptase PCR products and Western blotting. IRF-1 DNA-binding activity was assessed by electrophoretic shift mobility assay. The effect of gliadin stimulation on IRF-1 induction was investigated in an ex vivo organ culture of treated CD biopsies. Enhanced IRF-1 was seen in untreated CD in comparison with controls. This was evident at both the RNA and protein level. Furthermore, untreated CD samples exhibited stronger nuclear accumulation and DNA-binding activity of IRF-1 than controls. In contrast, IRF-2, a transcriptional repressor that binds the same DNA element and competes with IRF-1, was expressed at the same level in nuclear proteins extracted from both untreated CD and control patients. In explant cultures of treated CD biopsies, gliadin enhanced both IRF-1 RNA and protein. This effect was prevented by a neutralizing IFN-gamma antibody. Furthermore, stimulation of normal duodenal biopsies with IFN-gamma enhanced IRF-1. These data indicate that IRF-1 is a hallmark of the gliadin-mediated inflammation in CD and suggest that IFN-gamma/IRF-1 signaling pathway can play a key role in maintaining and expanding the local Th1 inflammatory response in this disease.

Rivista: PEDIATRIC RESEARCH

anno: 2003

volume n.: 54(3)

pag. da: 312

a:318

Macrolinea Cod.: 076.ISA.03

Autori: Schlosser G, Pocsfalvi G, Malorni A, Puerta A, de Frutos M, Vekey K.

Autori CNR:  GABRIELLA KATALIN POCSFALVI;  ANTONIO MALORNI; 

Titolo: Detection of immune complexes by matrix-assisted laser desorption/ionization mass spectrometry.

Abstract:

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used to detect an immune complex formed between beta-lactoglobulin and polyclonal anti-beta-lactoglobulin antibody in the gas phase. The most important experimental parameters to detect such a specific antibody-antigen complex by MALDI were the use of solutions at near-neutral pH and of sinapinic acid matrix prepared by the dried-droplet method. Under such conditions, predominantly one but also two molecules of antigen protein were complexed by the antibody. Specific formation of the antibody-antigen complex was confirmed by performing competitive reactions. Addition of antibody to a 1:1 mixture of beta-lactoglobulin and one control protein resulted not only in the appearance of the expected antibody-antigen complex, but also in a strong decrease in the free beta-lactoglobulin signal, while the abundance of the control protein was not influenced. Copyright 2003 John Wiley & Sons, Ltd.

Rivista: RAPID COMMUNICATIONS IN MASS SPECTROMETRY

anno: 2003

volume n.: 17

pag. da: 2741

a:2747

Macrolinea Cod.: 076.ISA.03

Autori: Schlosser G, Takats Z, Vekey K, Pocsfalvi G, Malorni A, Windberg E, Kiss A, Hudecz F.

Autori CNR:  GABRIELLA KATALIN POCSFALVI;  ANTONIO MALORNI; 

Titolo: Mass spectrometric analysis of combinatorial peptide libraries derived from the tandem repeat unit of MUC2 mucin.

Abstract:

Four 19-member synthetic peptide libraries, based on the TX1TX2T epitope motif of the mucin-2 gastrointestinal glycoprotein (MUC2) and ranging in peptide length from dipeptides to 15-mers (XT, TXT, TQTXT and KVTPTPTPTGTQTXT), were synthesized by combinatorial solid phase peptide synthesis using the portioning-mixing combinatorial approach, and analysed by electrospray ionization mass spectrometry at different (1000-10000) resolutions. Most of the components of the individual libraries could be easily identified in a single-stage molecular mass screening experiment. The resolving power of the instrument becomes an important factor above 800-1000 Da molecular mass, when predominantly multiply charged molecular ions are formed. Approaches to the identification of isobars (glutamine/lysine), isomers leucine/isoleucine) and sequence variations by tandem mass spectrometry, and/or by high-performance liquid chromatography-mass spectrometry are outlined.

Rivista: JOURNAL OF PEPTIDE SCIENCE

anno: 2003

volume n.: 9(6):

pag. da: 361

a:374

Macrolinea Cod.: 076.ISA.02

Autori: Strazzullo P, Galletti F, Barba G.

Autori CNR:  GIANVINCENZO BARBA; 

Titolo: Altered renal handling of sodium in human hypertension: short review of the evidence

Abstract:

A pathogenic role of the kidney in hypertension has been strongly supported by experimental studies by Guyton and Dahl since the 1960s. In the early 1980s, de Wardener and MacGregor proposed that in hypertensive patients the ability of the kidneys to excrete a sodium load could be genetically impaired. Since then, "sodium-sensitive" hypertension has been the object of numerous studies, mostly on animal models because of the difficulty to investigate the renal handling of sodium in humans. More recently, considerable progress in this field has been made thanks to the in vivo study of segmental renal tubular function by the clearance of lithium and to the growing knowledge of the genetics of renal tubular sodium transport systems. The scope of this review is to briefly review the most relevant information gathered by the investigation of segmental renal tubular sodium handling in humans as related to blood pressure regulation and hypertension. In aggregate, the results of these studies strongly support the association between altered renal sodium handling and high blood pressure and suggest a causal role of genetic, nutritional, metabolic, and neurohormonal factors. All of these factors, alone or in combination, may be able to impair the normal renal tubular sodium handling and influence blood pressure homeostasis. The paradigm of the pathogenic role of the kidney in hypertension is thus relentlessly shifting toward the definition of inherited as well as acquired renal tubular defects and molecular alterations, providing a plausible explanation for the alteration in blood pressure levels.

Rivista: HYPERTENSION

anno: 2003

volume n.: 41

pag. da: 1000

a:1005

Macrolinea Cod.: 076.ISA.02

Autori: Strazzullo P, Iacone R, Iacoviello L, Russo O, Barba G, Russo P, D'Orazio A, Barbato A, Cappuccio FP, Farinaro E, Siani A.

Autori CNR:  ALFONSO SIANI;  PAOLA RUSSO;  GIANVINCENZO BARBA; 

Titolo: Genetic variation in the renin-angiotensin system and abdominal adiposity in men: the Olivetti Prospective Heart Study

Abstract:

BACKGROUND: The renin-angiotensin system is involved in adipocyte growth and differentiation and possibly in adipose tissue metabolism. OBJECTIVE: To investigate the association of polymorphism in the angiotensin-converting enzyme (ACE) I/D gene, angiotensinogen M235T gene, and angiotensin II type 1 receptor A1166C gene with body mass index, body fat pattern, and obesity-associated hypertension. DESIGN: Cross-sectional longitudinal study. SETTING: The Olivetti factories in Marcianise and Pozzuoli, suburbs of Naples, Italy. PARTICIPANTS: 959 adult men, 25 to 75 years of age. MEASUREMENTS: Renin-angiotensin system polymorphism, anthropometric indexes, blood pressure, and serum glucose and insulin levels. RESULTS: No association was detected between angiotensinogen or angiotensin II type 1 receptor gene polymorphism and anthropometric indexes or blood pressure. For ACE I/D polymorphism, significant age-genotype interaction was detected on cross-sectional observation; the relation of body mass index, waist circumference, and diastolic blood pressure to age was significantly greater in persons with the DD genotype than in those with the ID or II genotype. Overweight and abdominal adiposity were more common in men with the DD genotype, particularly among older participants (51.1% vs. 36.5% and 33.1% vs. 22.0%, respectively). Odds ratios were 1.82 (95% CI, 1.16 to 2.87) for overweight and 1.76 (CI, 1.06 to 2.90) for abdominal adiposity. Among 314 untreated men first examined 20 years earlier, those with the DD genotype had greater age-adjusted weight gain (1.45 kg [CI, 0.12 to 2.78 kg]) and change in diastolic blood pressure (2.83 mm Hg [CI, 0.39 to 5.28 mm Hg]). The relative risk for overweight was 2.34 (CI, 1.32 to 4.15) among participants with the DD genotype versus those with the ID or II genotype. CONCLUSIONS: The ACE I/D polymorphism was a significant predictor of overweight and abdominal adiposity in men. DD homozygosity was associated with larger increases in body weight and blood pressure in aging persons, as well as with higher incidence of overweight.

Rivista: ANNALS OF INTERNAL MEDICINE

anno: 2003

volume n.: 138

pag. da: 17

a:23

Macrolinea Cod.: 076.ISA.03

Autori: Troncone R, Franzese A, Mazzarella G, Paparo F, Auricchio R, Coto I, Mayer M, Greco L.

Autori CNR:  GIUSEPPE MAZZARELLA; 

Titolo: Gluten sensitivity in a subset of children with insulin dependent diabetes mellitus.

Abstract:

OBJECTIVES: The association between celiac disease and insulin dependent diabetes mellitus (IDDM) is well established. Rectal gluten challenge has been used in patients with celiac disease and in first degree relatives as a tool to assess the mucosal immune response to gluten. The aim of this study was to assess the mucosal immune response to gluten in IDDM children by rectal gluten challenge. METHODS: Rectal biopsy specimens were obtained from 19 children with IDDM before and 6 h after rectal challenge with 2 g of a peptic tryptic digest of gliadin. A total of 16 treated celiac patients and 10 control subjects were also investigated. Epithelium and lamina propria CD3(+) and gamma delta(+) lymphocytes were counted with reference to a standard reference area of muscularis mucosae (10(4) microm(2)). RESULTS: After a local instillation of gliadin, a significant (>mean + 1 SD) percentage increment of lamina propria and epithelium CD3(+) and of lamina propria and epithelium gamma delta(+) lymphocytes was observed in five IDDM children, as compared to 11 and 13 celiac patients and one and two controls, respectively. A discriminant analysis allowed correct classification of 100% of patients with celiac disease and controls. The same analysis classified four of 19 IDDM children in the group of celiac patients. The positivity was associated with normal serology (antigliadin antibody, antiendomysial antibody, and antitissue transglutaminase antibodies) and a morphologically normal jejunal mucosa. All four patients had HLA-DQ alleles associated with celiac disease. CONCLUSIONS: Approximately 20% of IDDM children react to rectal instillation of gliadin. Long term follow-up is necessary to establish whether these subjects are at increased risk for developing celiac disease.

Rivista: AMERICAN JOURNAL OF GASTROENTEROLOGY

anno: 2003

volume n.: 98-3

pag. da: 590

a:595