Design of an epithermal column for BNCT based on D–D fusion neutron facility

Abstract Boron Neutron Capture Therapy (BNCT) is currently performed on patients at nuclear reactors. At the same time the international BNCT community is engaged in the development of alternative facilities for in-hospital treatments. This paper investigates the potential of a novel high-output D–D neutron generator, developed at Lawrence Berkeley National Laboratory (CA, USA), for BNCT. The simulation code MCNP-4C is used to realize an accurate study of the epithermal column in view of the treatment of deep tumours. Different materials and Beam Shaping Assemblies (BSA) are investigated and an optimized configuration is proposed. The neutron beam quality is defined by the standard free beam parameters, calculated averaging over the collimator aperture. The results are discussed and compared with the performances of other facilities

Age determination and authentication of ceramics: advancements in the thermoluminescence dating laboratory in Torino (Italy)

13Classified as an absolute dating method, thermoluminescence (TL) is a well-established radiation-based technique for the age determination and authentication of ceramic materials. Specifically, this method allows the determination of the time elapsed since kiln firing (or later fire events) by evaluating the luminescent emission of ceramics under heating at high temperatures. This paper provides a comprehensive presentation of the TL laboratory developed over the last decade at the Physics Department of the University of Torino. The laboratory was set up in collaboration with TecnArt S.r.l. and is also currently operating within the cultural heritage network of the National Institute of Nuclear Physics (INFN-CHNet). More than 10 years of experience in the field has resulted in improvements in procedures, with the development of customised alpha- and beta-irradiation systems and the optimisation of sampling approaches and chemical pre-treatment. Thanks to TecnArt S.r.l., the laboratory has been employed for dating and authenticating hundreds of archaeological sites and artworks, some of which are discussed in this work and compared, when possible, with radiocarbon dating.openopenGuidorzi, Laura; Fantino, Fulvio; Durisi, Elisabetta; Ferrero, Marco; Re, Alessandro; Vigorelli, Luisa; Visca, Lorenzo; Gulmini, Monica; Dughera, Giovanni; Giraudo, Giuseppe; Angelici, Debora; Panero, Elisa; Lo Giudice, AlessandroGuidorzi, Laura; Fantino, Fulvio; Durisi, Elisabetta; Ferrero, Marco; Re, Alessandro; Vigorelli, Luisa; Visca, Lorenzo; Gulmini, Monica; Dughera, Giovanni; Giraudo, Giuseppe; Angelici, Debora; Panero, Elisa; Lo Giudice, Alessandr

X-ray tomography of large wooden artworks: The case study of "Doppio corpo" by Pietro Piffetti

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RsaL-driven negative regulation promotes heterogeneity in Pseudomonas aeruginosa quorum sensing

In its canonical interpretation, quorum sensing (QS) allows single cells in a bacterial population to synchronize gene expression and hence perform specific tasks collectively once the quorum cell density is reached. However, growing evidence in different bacterial species indicates that considerable cell-to-cell variation in the QS activation state occurs during growth, often resulting in coexisting subpopulations of cells in which QS is active (quorate cells) or inactive (non-quorate cells). Heterogeneity has been observed in the las QS system of the opportunistic pathogen Pseudomonas aeruginosa. However, the molecular mechanisms underlying this phenomenon have not yet been defined. The las QS system consists of an incoherent feedforward loop in which the LasR transcriptional regulator activates the expression of the lasI synthase gene and rsaL, coding for the lasI transcriptional repressor RsaL. Here, single-cell-level gene expression analyses performed in ad hoc engineered biosensor strains and deletion mutants revealed that direct binding of RsaL to the lasI promoter region increases heterogeneous activation of the las QS system. Experiments performed with a dual-fluorescence reporter system showed that the LasR-dependent expression of lasI and rsaL does not correlate in single cells, indicating that RsaL acts as a brake that stochastically limits the transition of non-quorate cells to the quorate state in a subpopulation of cells expressing high levels of this negative regulator. Interestingly, the rhl QS system that is not controlled by an analogous RsaL protein showed higher homogeneity with respect to the las system

A GFP-lacZ Bicistronic Reporter System for Promoter Analysis in Environmental Gram-Negative Bacteria

Here, we describe a bicistronic reporter system for the analysis of promoter activity in a variety of Gram-negative bacteria at both the population and single-cell levels. This synthetic genetic tool utilizes an artificial operon comprising the gfp and lacZ genes that are assembled in a suicide vector, which is integrated at specific sites within the chromosome of the target bacterium, thereby creating a monocopy reporter system. This tool was instrumental for the complete in vivo characterization of two promoters, Pb and Pc, that drive the expression of the benzoate and catechol degradation pathways, respectively, of the soil bacterium Pseudomonas putida KT2440. The parameterization of these promoters in a population (using β-galactosidase assays) and in single cells (using flow cytometry) was necessary to examine the basic numerical features of these systems, such as the basal and maximal levels and the induction kinetics in response to an inducer (benzoate). Remarkably, GFP afforded a view of the process at a much higher resolution compared with standard lacZ tests; changes in fluorescence faithfully reflected variations in the transcriptional regimes of individual bacteria. The broad host range of the vector/reporter platform is an asset for the characterization of promoters in different bacteria, thereby expanding the diversity of genomic chasses amenable to Synthetic Biology methods

A Novel Extracytoplasmic Function (ECF) Sigma Factor Regulates Virulence in Pseudomonas aeruginosa

Next to the two-component and quorum sensing systems, cell-surface signaling (CSS) has been recently identified as an important regulatory system in Pseudomonas aeruginosa. CSS systems sense signals from outside the cell and transmit them into the cytoplasm. They generally consist of a TonB-dependent outer membrane receptor, a sigma factor regulator (or anti-sigma factor) in the cytoplasmic membrane, and an extracytoplasmic function (ECF) sigma factor. Upon perception of the extracellular signal by the receptor the ECF sigma factor is activated and promotes the transcription of a specific set of gene(s). Although most P. aeruginosa CSS systems are involved in the regulation of iron uptake, we have identified a novel system involved in the regulation of virulence. This CSS system, which has been designated PUMA3, has a number of unusual characteristics. The most obvious difference is the receptor component which is considerably smaller than that of other CSS outer membrane receptors and lacks a β-barrel domain. Homology modeling of PA0674 shows that this receptor is predicted to be a bilobal protein, with an N-terminal domain that resembles the N-terminal periplasmic signaling domain of CSS receptors, and a C-terminal domain that resembles the periplasmic C-terminal domains of the TolA/TonB proteins. Furthermore, the sigma factor regulator both inhibits the function of the ECF sigma factor and is required for its activity. By microarray analysis we show that PUMA3 regulates the expression of a number of genes encoding potential virulence factors, including a two-partner secretion (TPS) system. Using zebrafish (Danio rerio) embryos as a host we have demonstrated that the P. aeruginosa PUMA3-induced strain is more virulent than the wild-type. PUMA3 represents the first CSS system dedicated to the transcriptional activation of virulence functions in a human pathogen

Manipulating virulence factor availability can have complex consequences for infections

Given the rise of bacterial resistance against antibiotics, we urgently need alternative strategies to fight infections. Some propose we should disarm rather than kill bacteria, through targeted disruption of their virulence factors. It is assumed that this approach (i) induces weak selection for resistance because it should only minimally impact bacterial fitness, and (ii) is specific, only interfering with the virulence factor in question. Given that pathogenicity emerges from complex interactions between pathogens, hosts and their environment, such assumptions may be unrealistic. To address this issue in a test case, we conducted experiments with the opportunistic human pathogen Pseudomonas aeruginosa, where we manipulated the availability of a virulence factor, the iron-scavenging pyoverdine, within the insect host Galleria mellonella. We observed that pyoverdine availability was not stringently predictive of virulence and affected bacterial fitness in nonlinear ways. We show that this complexity could partly arise because pyoverdine availability affects host responses and alters the expression of regulatorily linked virulence factors. Our results reveal that virulence factor manipulation feeds back on pathogen and host behaviour, which in turn affects virulence. Our findings highlight that realizing effective and evolutionarily robust antivirulence therapies will ultimately require deeper engagement with the intrinsic complexity of host-pathogen systems