R. Papi

AbstractJournal URL: http://www3.interscience.wiley.com/journal/119877016/tocgroupBrucella spp. are gram-negative intercellular pathogens that infect a variety of domestic and wild animals. Brucella melitensis causes brucellosis to sheep, cattle and humans as a secondary host. It continues to be of great health significance and economic importance in many European countries. Differentiation between species and their different biovars is currently based on serotyping, phage typing, biochemical characteristics, metabolic properties and dye-sensitivity. The finger printing method was implemented using microsatellite DNA to genotype B. melitensis. An eight-base pair tandem repeat sequence was discovered in eight genomic loci of the B. melitensis genome and it was found that existed in various, multiple and accidental repetitions. Specific primers were used to amplify these particular loci. The PCR products were analysed in LI-COR IR2 DNA Sequencer and their molecular size was determined...

Journal URL: http://www.springer.at/amino_acidsThe production of molecularly imprinted polymers (MIPs) for the recognition of C-terminal cholecystokinin pentapeptide (CCK-5P) in the presence or absence of metal ions is reported. Metal ions play an important role in recognition process allowing the monomers to assemble around the template in protic solvents by ligand formation. The MIPs were prepared under the same molar ratio of template to monomers (acrylamide N N0-methylene bisacrylamide) in the presence or absence of nitriloacetic acid nickel complex (Ni-NTA). Subsequently the produced Ni-NTA polymers were washed with EDTA (pH 8) to remove the Ni2ώ and incubated in aqueous solution of MgSO4 FeSO4 ZnSO4 CoSO4 or CuSO4 respectively at initial concentration 20mM in order to examine the contribution of these ions to recognition process. From the rebinding data a Langmuir adsorption isotherm was obtained for each metal ion used and the subsequent Scatchard plot analysis showed that th...

Journal URL: http://www.springer.at/amino_acidsAbstractIn this study site-directed mutagenesis was performed on coagulation factor IX (fIX) gene and their effect on the protein’s proteolytic activity was studied. Blood coagulation factor IX (fIX) is a vitamin K-dependent plasma serine protease that plays a major role in coagulation mechanism and haemostasis. It is synthesized in hepatocytes as a precursor protein of 461 amino acids. After several post-translational modifications the mature protein is secreted in the circulation as inactive zymogen. The zymogen is then activated during the coagulation cascade into an active serine protease. A truncated form of fIX gene (rf9) inserted in a pET22b vector was used in our experiments so as to be able to overexpress the protein in prokaryotic cells. Two mutations were induced on this gene. Both of them are described in databases as polymorphisms. The first one (A21975G) is on the protein’s activation peptide and causes the substitution of...

Journal URL: http://www.springerlink.com/content/104405/AbstractSerine protease inhibitors (Serpins) is a group of proteins with similar structures, which were first identified as a set of proteins able to inhibit proteases function. The human plasma proteins antithrombin and antitrypsin, which play key roles in controlling blood coagulation and inflammation, respectively, were the first members of the serpins superfamily to be extensively studied. Trypsin-like serine proteases are essential for many biological processes. Because of this a large number of synthetic peptides have been designed and synthesized, based on the structure of inhibitors, active against trypsin or chymotrypsin. In the present work we have synthesized a series of Na-hydrazinopeptoids and aza-peptoids and studied their trypsin inhibitory effect. These peptidomimetics are expected to show enhanced metabolitic stability and bioavailability in comparison with natural parent peptidic analogs. All the syntheses wer...

Journal URL: http://www.springer.at/amino_acidsMolecularly imprinted polymers (MIPs) are smart tailored-made materials used for the sensitive and selective recognition of small molecules and=or biologically important substances. The target molecule acting as a molecular template is copolymerized with an excess of afunctional monomer and a cross-linker. The template is entrapped in the formed polymer and following its removal complementary cavities in structure and in properties to the template are formed in the polymer. Consequently the polymer is able to rebind the template molecule with high affinity and specificity. The chemical and physical stability of MIPs in both aqueous and organic solvents their resistance at extreme pH and temperature conditions and their reusability revealed their promising applications in the field of biocatalysis diagnostics immunoassays bioseparations biomimetic sensors affinity chromatography while imprinting molecules of biological origin such as ami...

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

An entry from the Cambridge Structural Database, the world's repository for small molecule crystal structures. The entry contains experimental data from a crystal diffraction study. The deposited dataset for this entry is freely available from the CCDC and typically includes 3D coordinates, cell parameters, space group, experimental conditions and quality measures.

Phosphatase and tensin homolog deleted on chromosome ten (PTEN) is one of the most frequently mutated human tumor suppressor genes, implicated in cell growth and survival and suppressing tumor formation. Loss of PTEN activity, either at the protein or genomic level, has been related to many primary and metastatic malignancies including breast cancer. The present study investigates the heterozygosity, mutation spectrum and protein expression of PTEN in 43 patients with breast cancer or precursor lesions of the breast and 10 healthy individuals. Microsatellite analysis at the PTEN locus using D10S215, D10S541 and D10S579 markers indicated that the observed heterozygosity (Ho) is lower than the expected heterozygosity (Hs) in benign and malignant breast disease. Mutational analysis in exons 1, 5, 7 and 9 of the PTEN gene revealed several mutations, most of which cause truncation of the PTEN protein and consequently loss of activity. Increased circulating levels of PTEN and phosphorylat...

ABSTRACT Molecularly imprinted polymers (MIPs) have been prepared from methacrylic acid (MAA) and ethylene glycol dimethacrylate (EGDMA) as the functional monomer and crosslinker, respectively, for cholecystokinin C terminal pentapeptide (CCK-5), and screened for their rebinding characteristics. They were produced with alterations in the molar ratio of template/monomer/ crosslinker and the percentage of net rebinding and the imprinting factor (IF) were used for the evaluation of their imprinting efficacy. The dissociation constant values determined in the case of the polymers with better characteristics, further confirmed the results above. Scanning electron microscopy (SEM) images of MIPs were also obtained in an attempt to correlate their characteristics with polymer's morphology. Rebinding experiments with CCK-5 and CCK-8 peptides in the case of the polymer that presented higher performance, with net rebinding values 11.9 and 9%, respectively, indicated that this polymer may be a good candidate for the analysis of CCKrelated peptides.

Introduction. Coagulation factor IX plays an important intermediate role in the activation of blood coagulation. It is located within the blood plasma as a zymogen, in its inactivated state. Factor IX is dependent on the presence of Vitamin K. The structure of factor IX closely resembles the structures of many other Vitamin K dependent plasma proteins, such as prothrombin, factor X and protein C. After being activated, Factor IX forms a complex with calcium ions, membrane phospholipids and coagulation factor VIIIa to activate factor X. The exact locus of the coagulation factor IX gene was found to exist in the Xq26-q27 region of the X chromosome. The FIX gene spans 34 kb and contains eight exons. Over 300 different mutations have been identified in the FIX gene, all of which result in the production of inactive FIX, causing hemophilia B. Aim. In this study we searched for mutations in the FIX gene which result in an increased activity of FIX thus being the cause of thrombophilia syn...

Present work aimed to examine the antibiotic resistance of the Salmonella enterica serotype Enteritidis (SE) isolates from poultry, to study the plasmid-mediated ampicillin resistance and to detect and determine the distribution of sodCI, sopE and sefA genes. Thirty-five SE isolates from one-day chicks, layers and broilers were studied for susceptibility/resistance to sixteen antimicrobial agents; 23 (65.7%) of them showed resistance to ampicillin, 5 (14.3%) to ampicillin and tetracycline, 4 (11.45%) to tetracycline and 1 (2.9%) isolate showed multi-drug resistance. Ampicillin (AmpR) and ampicillin/tetracycline (AmpRTeR) resistance was easily transferred by conjugation, and all isolates except two possessed a common band. The molecular mass of the plasmid carrying ampicillin resistance was approximately determined at 41kb after DNA digestion with BamHI, HindIII, EcoRI, EcoRV and PstI restriction enzymes and ligation of EcoRI fragments to pET29c. For the detection of TEM-1 or/and TEM...

The involvement of histamine and H4 receptor (H4 R) in cancer has been investigated recently using the H4 R agonists and antagonists. The scope of the research project was synthesis and exploration of the consequences of a group of compounds with histamine H4 receptor (H4 R) affinity on the promoter of PTEN gene encoding the antitumor PTEN protein. The series of novel compounds based either on H4 R antagonists JNJ7777120 structure or 1,3,5-triazine scaffold were synthesized, evaluated for histamine H4 R affinity and used in this study. Compounds 5 and 7 belonging to the group of JNJ7777120 analogues showed the highest interaction with the promoter of PTEN gene and weak affinity against H4 R with Ki value >100 μm. These compounds showed no significant effect on neuroblastoma IMR-32 cells viability indicating no correlation between PTEN gene promoter affinity and antitumor activity. Compound 6, another JNJ7777120 analogue, showed the highest effect on IMR-32 viability with calculated IC50 = 23.27 μm. The 1,3,5-triazine derivatives exhibited generally low or medium interaction with PTEN gene promoter. However, the 1,3,5-triazine derivative 11 with the para-bromo substituent showed the highest affinity against H4 R with Ki value of 520 nm and may be considered as a new lead structure.

A new series of coordination compounds of the starting materials [Cu(dienX(2)Y(2))] and their adducts [Cu(dienXXY(2))(2a-5mt)] (where dien=diethylenetriamine, dienXX=Schiff bases of diethylenetriamine with 2-furaldehyde or 2-thiophene-carboxaldehyde, X=O, S, Y=Cl, Br, NO(3) and 2a-5mt=2-amino-5-methylthiazole) were synthesized by stepwise reactions and their structures were established by C, H, N, Cu analysis, spectroscopic, magnetic and molar conductivity measurements. The isolated compounds are monomers, paramagnetic and electrolytic compounds of the type 1:1. In all cases, the pentadentate Schiff base (dienXX) is bonded in a tridentate fashion through the 3 N atoms. In the CudienXXY(2) compounds the coordination sphere is completed by two Cl or Br or NO(3) groups in a square pyramidal arrangement. The proposed structure for this type of compound was further supported by X-ray diffraction analysis of the compound [Cu(dienOO)Cl(2)]. Its basal plane consists of three Cu-N contacts [2.017(2), 2.025(2) and 2.012(2) A] from dienOO, and the Cl(1) atom, while the Cl(2) atom possesses the apical position, the relevant distances being 2.2732(7) A for Cu-Cl(1) and 2.6051(7) A Cu-Cl(2). In the CudienX(2)Y(2).2a-5mt adducts the coordination sphere of copper is further completed by the nitrogen ring atom of the 2a-5mt, forming an octahedral configuration. The study of the biological activity of the compounds synthesized against a panel of different normal and cancer cell lines (MRC5, HeLa, MCF7, HT-29, OAW42, T47D) and bacteria (E. coli, B. cereus, B. subtilis) showed that the adducts of the type [Cu(dienXXY(2))(2a-5mt)] exhibit increased activity both in cancer cells and in bacteria, compared to the starting material of type [Cu(dienXXY(2))].

Assessment of genetic variation within and among populations is an essential parameter for the effective conservation of forest genetic resources. In this work, the genetic diversity within and among natural populations of the forest tree species Fraxinus angustifolia Vahl in Greece was studied using selected nuclear and chloroplast microsatellite DNA loci. Eight natural populations of F. angustifolia were identified in different locations of the mainland of Greece, and a total of 230 individuals were studied. High polymorphism was observed within populations, while the genetic differentiation among populations was moderate. Intra-population diversity was correlated with geographical coordinates, but no isolation by distance was observed. Of the three haplotypes identified, only one was dominant. Putative ancestral haplotypes were found at small spatial scales suggesting that population expansions could have originated in the region. This study located in sympatry haplotypes that in...