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Zummo F.P.,University of Palermo | Marineo S.,Istituto Zooprofilattico Sperimentale della Sicilia A. Mirri | Pace A.,University of Palermo | Civiletti F.,University of Turin | And 2 more authors.
Applied Microbiology and Biotechnology

Most enzymes involved in tryptophan catabolism via kynurenine formation are highly conserved in Prokaryotes and Eukaryotes. In humans, alterations of this pathway have been related to different pathologies mainly involving the central nervous system. In Bacteria, tryptophan and some of its derivates are important antibiotic precursors. Tryptophan degradation via kynurenine formation involves two different pathways: the eukaryotic kynurenine pathway, also recently found in some bacteria, and the tryptophan-to-anthranilate pathway, which is widespread in microorganisms. The latter produces anthranilate using three enzymes also involved in the kynurenine pathway: tryptophan 2,3-dioxygenase (TDO), kynureninase (KYN), and kynurenine formamidase (KFA). In Streptomyces coelicolor, where it had not been demonstrated which genes code for these enzymes, tryptophan seems to be important for the calcium- dependent antibiotic (CDA) production. In this study, we describe three adjacent genes of S. coelicolor (SCO3644, SCO3645, and SCO3646), demonstrating their involvement in the tryptophan-to-anthranilate pathway: SCO3644 codes for a KFA, SCO3645 for a KYN and SCO3646 for a TDO. Therefore, these genes can be considered as homologous respectively to kynB, kynU, and kynA of other microorganisms and belong to a constitutive catabolic pathway in S. coelicolor, which expression increases during the stationary phase of a culture grown in the presence of tryptophan. Moreover, the S. coelicolor ΔkynU strain, in which SCO3645 gene is deleted, produces higher amounts of CDA compared to the wild-type strain. Overall, these results describe a pathway, which is used by S. coelicolor to catabolize tryptophan and that could be inactivated to increase antibiotic production. © Springer-Verlag 2012. Source

Licciardi M.,University of Palermo | Scialabba C.,University of Palermo | Fiorica C.,University of Palermo | Cavallaro G.,University of Palermo | And 3 more authors.
Molecular Pharmaceutics

In this paper the preparation of magnetic nanocarriers (MNCs), containing superparamagnetic domains, is reported, useful as potential magnetically targeted drug delivery systems. The preparation of MNCs was performed by using the PHEA-IB-p(BMA) graft copolymer as coating material through the homogenization-solvent evaporation method. Magnetic and nonmagnetic nanocarriers containing flutamide (FLU-MNCs) were prepared. The prepared nanocarriers have been exhaustively characterized by dynamic light scattering (DLS), transmission electron microscopy (TEM), and magnetic measurements. Biological evaluation was performed by in vitro cytotoxicity and cell uptake tests and in vivo biodistribution studies. Magnetic nanocarriers showed dimensions of about 300 nm with a narrow size distribution, an amount of loaded FLU of 20% (w/w), and a superparamagnetic behavior. Cell culture experiments performed on prostate cancer cell line LNCaP demonstrated the cytotoxic effect of FLU-MNCs. In vivo biodistribution studies carried out by the application of an external magnetic field in rats demonstrated the effect of the external magnet on modifying the biodistribution of FLU-MNCs. FLU-MNCs resulted efficiently internalized by tumor cells and susceptible to magnetic targeting by application of an external magnetic field. The proposed nanocarriers can represent a very promising approach to obtain an efficient magnetically targeted anticancer drug delivery system. © 2013 American Chemical Society. Source

Cristani M.,Messina University | Naccari C.,Messina University | Nostro A.,Messina University | Pizzimenti A.,Istituto Zooprofilattico Sperimentale della Sicilia A. Mirri | And 2 more authors.
Environmental Science and Pollution Research

Background, aim and scope Metal pollution is a serious problem for environmental safety and programmes of monitoring and bioremediation are needed. Among the processes of bioremediation, the use of microbes to remove and degrade contaminants is considered a biotechnological approach to clean up polluted environments. Aim The aim of this study was to evaluate the ability of Serratia marcescens in Pb, Cd and Cr removal and the potential use of these bacteria in toxic metal bioremediation from polluted environments. Methods A short-term study (120 min) was carried out to study the bacterial growth in the presence of sub-inhibitory concentrations of each metal analysed and the kinetics of metal biosorption in S. marcescens strain. In addition, metal influence on the biosynthesis of the red pigment 'prodigiosina' by S. marcescens was monitored. Results: The results obtained in this study show metals biosorption by S. marcescens (range: 0.0133-0.213 μg/g for Pb; 0.097-0.1853 μg/g for Cd; and 0.105-0.176 μg/g for Cr) and confirm the possible use of this bacterium to realize bioremediation processes, especially for Pb removal, and as a bioindicator of metal pollution. © 2011 Springer-Verlag. Source

Schillaci D.,University of Palermo | Cusimano M.G.,University of Palermo | Cunsolo V.,University of Catania | Saletti R.,University of Catania | And 4 more authors.
AMB Express

The present study aims to investigate coelomocytes, immune mediators cells in the echinoderm Holothuria tubulosa, as an unusual source of antimicrobial and antibiofilm agents. The activity of the 5kDa peptide fraction of the cytosol from H. tubulosa coelomocytes (5-HCC) was tested against a reference group of Gram-negative and Gram-positive human pathogens. Minimal inhibitory concentrations (MICs) ranging from 125 to 500 mg/ml were determined against tested strains. The observed biological activity of 5-HCC could be due to two novel peptides, identified by capillary RP-HPLC/nESI-MS/MS, which present the common chemical-physical characteristics of antimicrobial peptides. Such peptides were chemically synthesized and their antimicrobial activity was tested. The synthetic peptides showed broad-spectrum activity at 12.5 mg/ml against the majority of the tested Gram-positive and Gram-negative strains, and they were also able to inhibit biofilm formation in a significant percentage at a concentration of 3.1 mg/ml against staphylococcal and Pseudomonas aeruginosa strains. The immune mediators in H. tubulosa are a source of novel antimicrobial peptides for the development of new agents against biofilm bacterial communities that are often intrinsically resistant to conventional antibiotics. © 2013 Bel-Rhlid et al. Source

Juyena N.S.,University of Padua | Vencato J.,University of Padua | Pasini G.,University of Padua | Vazzana I.,Istituto Zooprofilattico Sperimentale della Sicilia A. Mirri | Stelletta C.,University of Padua
Reproduction, Fertility and Development

The aim of the present study was to evaluate the biochemical composition of seminal plasma, along with semen quality, of alpacas maintained on different diets (hay; hay+pasture grazing; pasture grazing+sheep concentrate; pasture grazing+horse concentrate; Periods 1-4, respectively). Alpacas (n≤5) were fed the four different diets for a period of 6 weeks each. During the period of feeding of each diet, semen was collected using an artificial vagina to determine its volume, viscosity, sperm concentration and sperm motility. Moreover, testicular volume and body condition score were evaluated. Seminal plasma was analysed biochemically to measure total protein, triglyceride, cholesterol, γ-glutamyl transferase, alanine aminotransferase (ALT) and alkaline phosphatase levels. Protein profiles were investigated using one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. There was high variability in semen parameters between different males maintained on the same diet. Semen volume increased significantly (P<0.05) when alpacas were fed diets containing commercial sheep and horse concentrates. In contrast, sperm concentration and motility decreased significantly (P<0.05) from Period 1 to Period 4. Dietary changes had no effect on viscosity. Significant reductions were seen in triglyceride and cholesterol content, as well as γ-glutamyl transferase, ALT and alkaline phosphatase concentrations, from Period 1 to Period 4. Regardless of experimental period, a wide variation was seen in seminal plasma enzyme concentrations between alpacas, whereas diet had no effect on glucose and total protein concentrations in the seminal plasma. Eight protein bands, with molecular weights ranging from 200 to 14kDa, were considered in electrophoresis gel after image analysis. Proteins fractions of the 14-kDa (total protein express in mddL-1 with a molecular weight of 14-kDa, TP8) and 21-kDa (total protein express in mddL-1 with a molecular weight of 21-kDa, TP7) bands were not present in all samples of alpaca seminal plasma. There were no significant changes in the concentration of any protein fractions during the four periods. Moreover, the protein fraction of the 60-kDa (total protein express in mddL-1 with a molecular weight of 60-kDa, TP3) band was the most prevalent in all periods. These results demonstrate that there are marked changes in semen quality, as well as some parameters related to the composition of alpaca seminal plasma, that are dependent on diet, which may indicate the need for specific diet formulation to improve reproductive performance. We hypothesise that, in alpacas, the mechanisms underlying the changes in some reproductive traits in response to feeding regimens could be related to changes in the endocrine-gonadal system. © 2013 CSIRO. Source

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