Antimicrobial susceptibility of vancomycin-susceptible and -resistant enterococci isolated in Italy from raw meat products, farm animals, and human infections

The susceptibility of vancomycin-resistant (VRE) and vancomycin-susceptible (VSE) enterococci to 10 antimicrobial agents was evaluated. The strains, belonging to different species, were isolated in Italy from raw meat products, farm animals, and human clinical infections in the years 1997-2000. High frequency of resistance to tetracycline and erythromycin was observed in all the groups of strains. On the contrary, chloramphenicol was the only drug that showed a relatively low rate of resistance in all the groups examined. In general, the resistance rates observed for VSE did not differ from those observed for VRE of the same species and origin. Some differences could be noticed among the different enterococcal species, with Enterococcus faecium strains being usually more resistant to beta-lactams, and Enterococcus faecalis strains more resistant to gentamicin. However, the strongest differences were observed when the strains were compared according to their source, the human isolates being usually more resistant than the isolates of animal origin. No significant difference was observed between isolates of swine and poultry origin. Among VRE E. faecium, multiple resistance was much more frequent among the human strains (90%) than among poultry (48.9%) and swine (26.5%) strains. These results show that in Italy VRE isolates from human clinical infections are usually more resistant than isolates from meat products and farm animals, and possess different antimicrobial resistance profiles.     

Enantioselective hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues using a penicillin G acylase-based HPLC monolithic silica column

A technique based on liquid chromatography has been developed to facilitate studies of enantioselectivity in penicillin G acylase (PGA)-catalyzed hydrolysis of some 2-aryloxyalkanoic acid methyl esters and isosteric analogues. PGA was covalently immobilized on an aminopropyl monolithic silica support to create an immobilized HPLC-enzyme reactor. Two sets of experimental data were drawn to calculate the enantioselectivity (E) of the kinetically controlled enantiomer-differentiating reaction, the degree of substrate conversion and the enantiomeric excess of the product. The developed enzymatic reactor was coupled through a switching valve to an achiral analytical column for separation and quantitation of the hydrolysis products. The enantiomeric excess was determined off-line on a PGA-chiral stationary phase. In this way, highly precise E values were determined. A computational study related to the hydrolysis of the considered racemic esters was also carried out in order to unambiguously clarify both the substrate specificity and the enantioselectivity displayed by PGA.     

Occupancy of the Dictyostelium cAMP receptor, cAR1, induces a reduction in affinity which depends upon COOH-terminal serine residues

Many G-protein-coupled receptors display a rapid decrease in ligand binding following pretreatment with agonist. cAR1, a cAMP receptor expressed early in the developmental program of Dictyostelium, mediates chemotaxis, activation of adenylyl cyclase, and gene expression changes that bring about the aggregation of 10(5) amoebae to form a multicellular structure. Occupancy of cAR1 by cAMP initiates multiple desensitization processes, one of which is an apparent reduction in binding sites. In transformed cells expressing cAR1 constitutively, Scatchard analyses revealed that this apparent loss of ligand binding is largely due to a significant reduction in the affinity of cAR1 for cAMP. A parallel increase in the dose dependence of cAR1-mediated cAMP uptake was observed. Consistent with these findings, proteolysis of intact cells and immunofluorescence suggested that cAR1 remains on the cell-surface following cAMP treatment. Finally, agonist-induced loss of ligand binding is impaired in cAR1 mutants lacking a cluster of cytoplasmic serine residues, which are targets of cAMP-induced phosphorylation.     

Biomimetic poly(glycerol sebacate) (PGS) membranes for cardiac patch application

In this study biomimetic poly(glycerol sebacate) PGS matrix was developed for cardiac patch application. The rationale was that such matrices would provide conducive environment for the seeded cells at the interphase with PGS. From the microstructural standpoint, PGS was fabricated into dense films and porous PGS scaffolds. From the biological aspect, biomimetic PGS membranes were developed via covalently binding peptides Tyr-Ile-Gly-Ser-Arg (YIGSR) and Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), corresponding to the epitope sequences of laminin and fibronectin, respectively onto the surface. To improve and enhance homogenous binding of peptides onto the PGS surface, chemical modification of its surface was carried out. A sequential regime of alkaline hydrolysis with 0.01 M NaOH for 5 min and acidification with 0.01 M HCl for 25 s was optimal. More COOH chemical group was exposed without causing deleterious effect on the bulk properties of the polymer as revealed by the physicochemical analysis carried out. HPLC analysis, chemical imaging and ToF-SIMS were able to establish the successful homogenous functionalization of PGS membranes with the peptides. Finally, the developed biomimetic membranes supported the adhesion and growth of rat and human cardiac progenitor cells.     

Strong-field phenomena driven by mid-infrared ultrafast sources

The development of ultrafast and intense laser sources has paved the way to the study of laser–matter interaction in the strong-field regime, where the amplitude of the laser field becomes comparable to the Coulomb field seen by electrons in the proximity of their parent ion. This realm embraces numerous phenomena, such as high-order harmonic generation, tunnel photoionization, laser-induced electron diffraction and holography, ponderomotive electron streaking, and so forth. For a long time, the Ti:Sapphire laser has been the sole workhorse of strong-field scientists; the recent development of intense and tunable ultrafast sources operating in the mid-infrared added an extra dimension to strong-field investigations, with valuable benefits in the understanding of several phenomena and in the development of high-photon-energy coherent sources. In this work we review the recent achievements in strong-field science which were allowed by the exploitation of mid-infrared ultrafast lasers.     

Microdosimetry for nanosecond pulsed electric field applications: a parametric study for a single cell

A microdosimetric study of nanosecond pulsed electric fields, including dielectric dispersivity of cell compartments, is proposed in our paper. A quasi-static solution based on the Laplace equation was adapted to wideband signals and used to address the problem of electric field estimation at cellular level. The electric solution was coupled with an asymptotic electroporation model able to predict membrane pore density. An initial result of our paper is the relevance of the dielectric dispersivity, providing evidence that both the transmembrane potential and the pore density are strongly influenced by the choice of modeling used. We note the crucial role played by the dielectric properties of the membrane that can greatly impact on the poration of the cell. This can partly explain the selective action reported on cancerous cells in mixed populations, if one considers that tumor cells may present different dielectric responses. Moreover, these kinds of studies can be useful to determine the appropriate setting of nsPEF generators as well as for the design and optimization of new-generation devices.     

Applicability of SSR markers to the traceability of monovarietal olive oils

BACKGROUND: To protect the features and authenticity of food products, the European Commission enforces two certification labels: Protected Designation of Origin (PDO) and Protected Geographical Indication (PGI). EEC Regulation No. 510/2006 imposes criteria for labelling, production and commercialisation of olive oil. Since plant genotype is a major determinant in establishing the PDO and PGI labels, methods to ascertain the varieties present in a batch of olive oil are essential in validating product conformity. The traceability of olive oil can be assessed through simple sequence repeat (SSR) co‐dominant markers targeted to specific regions of DNA from olive cultivars. RESULTS: Twenty‐one monovarietal olive oils were analysed with nine nuclear and two shortened SSRs. For each marker the correspondence of allelic profile with the reference cultivar, the reproducibility of profiles in different DNA extractions and the polymorphism information content were determined. CONCLUSION: The results showed that using a panel of SSR markers such as those described in this paper allows one to make a reliable attribution of an olive oil to a specific cultivar. Copyright © 2011 Society of Chemical Industry     

A classification tool for N-way array based on SIMCA methodology

In the literature there are only few papers concerned with classification methods for multi-way arrays. The most common procedure, by far, is to unfold the multi-way data array into an ordinary matrix and then to apply the traditional multivariate tools for classification. As opposed to unfolding the data several possibilities exist for building classification models more directly based on the multi-way structure of the data. As an example, multi-way partial least squares discriminant analysis has been used as a supervised classification method, another alternative that has been investigated is to perform classification using Fisher's LDA or SIMCA on the score matrix from e.g. a PARAFAC or a Tucker model. Despite a few attempts of applying such multi-way classification approaches, no-one has looked into how such models are best built and implemented.In this work, the SIMCA method is extended to three-way arrays. Included in this work is also actual code that will work on general multi-way arrays rather than just three-way arrays. In analogy with two-way SIMCA, a decomposition model is separately built for the multi-way data for each class, using multi-way decomposition method such as PARAFAC or Tucker3. In the choice of the best class dimensionality, i.e. number of latent factors, both the results of cross-validation but mainly the sensitivity/specificity values are evaluated. In order to estimate the class limits for each class model, orthogonal and score distances are considered, and different statistics are implemented and tested to set confidence limits for these two parameters. Classification performance using different definitions of class boundaries and classification rules, including the use of cross-validated residuals and scores is compared.The proposed N-SIMCA methodology and code, besides simulated data sets of varying dimensionality, has been tested on two case studies, concerning food authentication tasks for typical food products.