Las Lunas, Yuncler (Toledo). Un depósito de materiales metálicos del Bronce Final en la Submeseta Sur de la Península Ibérica

Se exponen los resultados del primer estudio realizado sobre un nuevo conjunto de materiales metálicos del Bronce Final recuperado a finales de 2008 en las excavaciones arqueológicas del yacimiento de Las Lunas (Yuncler, Toledo, España). La localización geográfica del hallazgo, lejos de las principales zonas de dispersión conocidas para este tipo de conjuntos, la singularidad de los objetos que integra, y las relaciones atlánticas y mediterráneas que evidencian sus materiales, lo convierten en un ejemplo destacado para el estudio de este período en el centro de la Península Ibérica.Peer reviewe

Detection and Quantification of Leptospira interrogans in Hamster and Rat Kidney Samples: Immunofluorescent Imprints versus Real-time PCR

A major limitation in the clinical management and experimental research of leptospirosis is the poor performance of the available methods for the direct detection of leptospires. In this study, we compared real-time PCR (qPCR), targeting the lipL32 gene, with the immunofluorescent imprint method (IM) for the detection and quantification of leptospires in kidney samples from the rat and hamster experimental models of leptospirosis. Using a virulent strain of Leptospira interrogans serovar Copenhageni, a chronic infection was established in the rat model, which were euthanized 28 days post-infection, while the hamster model simulated an acute infection and the hamsters were euthanized eight days after inoculation. Leptospires in the kidney samples were detected using culture isolation, qPCR and the IM, and quantified using qPCR and the IM. In both the acute and chronic infection models, the correlation between quantification by qPCR and the IM was found to be positive and statistically significant (P<0.05). Therefore, this study demonstrates that the IM is a viable alternative for not only the detection but also the quantification of leptospires, particularly when the use of qPCR is not feasible

Impact du vagabondage mental et de la mindfulness sur le continuum des symptômes TDAH à l'adolescence et à l'âge adulte dans une population sans diagnostic

Le trouble du déficit de l'attention avec ou sans hyperactivité (TDAH) est un trouble neurodéveloppemental reconnu par la 5ème édition du Diagnostic and Statistical Manual of Mental Disorders (DSM-V ; American Psychiatric Association, 2013), dont les principaux symptômes sont l'inattention et l'hyperactivité/impulsivité (American Psychiatric Association, 2013). Ces difficultés d'inattention et d'hyperactivité/impulsivité peuvent également être présentes à divers degrés dans la population générale et peuvent être handicapantes à l'adolescence ainsi qu'à l'âge adulte. Il est important d'investiguer les processus sous-jacents afin de mieux comprendre le fonctionnement psychologique dans ces catégories d'âge. Cette recherche a investigué les relations entre le vagabondage mental et la mindfulness, mais aussi la contribution de ces deux variables au continuum de symptômes TDAH dans la population générale. L'un des principaux résultats de cette recherche est que le vagabondage mental contribue à expliquer le continuum de symptômes TDAH chez les adolescents et les adultes sans diagnostic

Post-translational Modification of LipL32 during <i>Leptospira interrogans</i> Infection

<div><p>Background</p><p>Leptospirosis, a re-emerging disease of global importance caused by pathogenic <i>Leptospira</i> spp., is considered the world's most widespread zoonotic disease. Rats serve as asymptomatic carriers of pathogenic <i>Leptospira</i> and are critical for disease spread. In such reservoir hosts, leptospires colonize the kidney, are shed in the urine, persist in fresh water and gain access to a new mammalian host through breaches in the skin.</p><p>Methodology/Principal Findings</p><p>Previous studies have provided evidence for post-translational modification (PTM) of leptospiral proteins. In the current study, we used proteomic analyses to determine the presence of PTMs on the highly abundant leptospiral protein, LipL32, from rat urine-isolated <i>L. interrogans</i> serovar Copenhageni compared to <i>in vitro</i>-grown organisms. We observed either acetylation or tri-methylation of lysine residues within multiple LipL32 peptides, including peptides corresponding to regions of LipL32 previously identified as epitopes. Intriguingly, the PTMs were unique to the LipL32 peptides originating from <i>in vivo</i> relative to <i>in vitro</i> grown leptospires. The identity of each modified lysine residue was confirmed by fragmentation pattern analysis of the peptide mass spectra. A synthetic peptide containing an identified tri-methylated lysine, which corresponds to a previously identified LipL32 epitope, demonstrated significantly reduced immunoreactivity with serum collected from leptospirosis patients compared to the peptide version lacking the tri-methylation. Further, a subset of the identified PTMs are in close proximity to the established calcium-binding and putative collagen-binding sites that have been identified within LipL32.</p><p>Conclusions/Significance</p><p>The exclusive detection of PTMs on lysine residues within LipL32 from <i>in vivo</i>-isolated <i>L. interrogans</i> implies that infection-generated modification of leptospiral proteins may have a biologically relevant function during the course of infection. Although definitive determination of the role of these PTMs must await further investigations, the reduced immune recognition of a modified LipL32 epitope suggests the intriguing possibility that LipL32 modification represents a novel mechanism of immune evasion within <i>Leptospira</i>.</p></div

LipL32 synthetic peptides generated for immune recognition studies.

<p>LipL32 synthetic peptides generated for immune recognition studies.</p

Modified lysine residues mapped onto the Ca²⁺ bound LipL32 structure [39].

<p><b>A.</b> Secondary structure and <b>B.</b> surface representations. The numbering system C¹-K²⁵³ corresponds to the mature LipL32 protein (lacking the 19 residue cleaved SPII signal sequence) established by Vivian <i>et al.</i><a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003280#pntd.0003280-Vivian1" target="_blank">[40]</a> and adopted by Tung <i>et al.</i><a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003280#pntd.0003280-Tung1" target="_blank">[39]</a>. Note, the Ca²⁺ binding site and predicted collagen binding surface in panel <b>B</b> are highlighted in yellow and frame Lys199. The figure was generated using PyMOL (Molecular Graphics System, Version 1.5.0.4 Schrödinger, LLC).</p

Leptospirosis patient serum reactivity against modified and unmodified versions of synthetic LipL32 peptides.

<p>Shown is the reactivity of pooled sera from laboratory-confirmed leptospirosis patients (black bars) and LipL32-specific polyclonal rabbit serum (white bars) to unmodified and tri-methylated (KMe³) versions of P1 and P2 peptides comprising experimentally-determined B-cell epitopes of LipL32 <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003280#pntd.0003280-Lottersberger1" target="_blank">[36]</a>. Two independent experiments were performed with reproducible results; the results from one representative experiment are shown. Error bars represent standard error of measurement from triplicate samples. For statistical analyses, the level of reactivity of patient sera against the unmodified version of P1 was compared to the level of reactivity against the P1 peptide containing a tri-methylation at position K¹⁵² (P1-K¹⁵²Me³) using the Student's two-tailed <i>t</i>-test (*<i>p</i><0.0001).</p

Two-dimensional gel electrophoresis (2DGE) of <i>Leptospira</i> whole proteome profiles indicating LipL32 protein spots analyzed by MS/MS.

<p>(<b>A</b>) 2DGE profile of <i>in vitro</i>-cultured <i>Leptospira</i> (IVCL) and (<b>B</b>) 2DGE profile of rat urine-isolated <i>Leptospira</i> (RUIL). Protein derived from whole cells were separated using immobilized pH gradient IPG strips (pH 3–10, non-linear) followed by SDS-PAGE. Circled protein spots corresponding to LipL32 were excised for analysis by LC-MS/MS.</p