Extended-Spectrum-Beta-Lactamases, AmpC Beta-Lactamases and Plasmid Mediated Quinolone Resistance in Klebsiella spp. from Companion Animals in Italy

We report the genetic characterization of 15 Klebsiella pneumoniae (KP) and 4 isolates of K. oxytoca (KO) from clinical cases in dogs and cats and showing extended-spectrum cephalosporin (ESC) resistance. Extended spectrum beta-lactamase (ESBL) and AmpC genes, plasmid-mediated quinolone resistance (PMQR) and co-resistances were investigated. Among KP isolates, ST101 clone was predominant (8/15, 53%), followed by ST15 (4/15, 27%). ST11 and ST340, belonging to Clonal Complex (CC)11, were detected in 2012 (3/15, 20%). MLST on KP isolates corresponded well with PFGE results, with 11 different PFGE patterns observed, including two clusters of two (ST340) and four (ST101) indistinguishable isolates, respectively. All isolates harbored at least one ESBL or AmpC gene, all carried on transferable plasmids (IncR, IncFII, IncI1, IncN), and 16/19 were positive for PMQR genes (qnr family or aac(6')-Ib-cr). The most frequent ESBL was CTX-M-15 (11/19, 58%), detected in all KP ST101, in one KP ST15 and in both KP ST340. blaCTX-M-15 was carried on IncR plasmids in all but one KP isolate. All KP ST15 isolates harbored different ESC resistance genes and different plasmids, and presented the non-transferable blaSHV-28 gene, in association with blaCTX-M-15, blaCTX-M-1 (on IncR, or on IncN), blaSHV-2a (on IncR) or blaCMY-2 genes (on IncI1). KO isolates were positive for blaCTX-M-9 gene (on IncHI2), or for the blaSHV-12 and blaDHA-1 genes (on IncL/M). They were all positive for qnr genes, and one also for the aac(6')-Ib-cr gene. All Klebsiella isolates showed multiresistance towards aminoglycosides, sulfonamides, tetracyclines, trimethoprim and amphenicols, mediated by strA/B, aadA2, aadB, ant (2")-Ia, aac(6')-Ib, sul, tet, dfr and cat genes in various combinations. The emergence in pets of multidrug-resistant Klebsiella with ESBL, AmpC and PMQR determinants, poses further and serious challenges in companion animal therapy and raise concerns for possible bi-directional transmission between pets and humans, especially at household level

Thermal Conductivity of Ordered Mesoporous Nanocrystalline Silicon Thin Films Made from Magnesium Reduction of Polymer-Templated Silica

This paper reports the cross-plane thermal conductivity of ordered mesoporous nanocrystalline silicon thin films between 25 and 315 K. The films were produced by evaporation induced self-assembly of mesoporous silica followed by magnesium reduction. The periodic ordering of pores in mesoporous silicon was characterized by X-ray diffraction and direct SEM imaging. The average crystallite size, porosity, and film thickness were about 13 nm, 25-35%, and 140-340 nm, respectively. The pores were arranged in a face-centered cubic lattice. The cross-plane thermal conductivity of the mesoporous silicon thin films was measured using the 3ω method. It was between 3 and 5 orders of magnitude smaller than that of bulk single crystal silicon in the temperature range considered. The effects of temperature, film thickness, and copolymer template on the thermal conductivity were investigated. A model based on kinetic theory was used to accurately predict the measured thermal conductivity for all temperatures. On the one hand, both the measured thermal conductivity and the model predictions showed a temperature dependence of k proportional to T2 at low temperatures, typical of amorphous and strongly disordered materials. On the other hand, at high temperatures the thermal conductivity of mesoporous silicon films reached a maximum, indicating a crystalline-like behavior. These results will be useful in designing mesoporous silicon with desired thermal conductivity by tuning its morphology for various applications

Complete Nucleotide Sequence of CTX-M-15-Plasmids from Clinical Escherichia coli Isolates: Insertional Events of Transposons and Insertion Sequences

BACKGROUND: CTX-M-producing Escherichia coli strains are regarded as major global pathogens. METHODOLOGY/PRINCIPAL FINDINGS: The nucleotide sequence of three plasmids (pEC_B24: 73801-bp; pEC_L8: 118525-bp and pEC_L46: 144871-bp) from Escherichia coli isolates obtained from patients with urinary tract infections and one plasmid (pEC_Bactec: 92970-bp) from an Escherichia coli strain isolated from the joint of a horse with arthritis were determined. Plasmid pEC_Bactec belongs to the IncI1 group and carries two resistance genes: bla(TEM-1) and bla(CTX-M-15). It shares more than 90% homology with a previously published bla(CTX-M)-plasmid from E. coli of human origin. Plasmid pEC_B24 belongs to the IncFII group whereas plasmids pEC_L8 and pEC_L46 represent a fusion of two replicons of type FII and FIA. On the pEC_B24 backbone, two resistance genes, bla(TEM-1) and bla(CTX-M-15), were found. Six resistance genes, bla(TEM-1), bla(CTX-M-15), bla(OXA-1), aac6'-lb-cr, tetA and catB4, were detected on the pEC_L8 backbone. The same antimicrobial drug resistance genes, with the exception of tetA, were also identified on the pEC_L46 backbone. Genome analysis of all 4 plasmids studied provides evidence of a seemingly frequent transposition event of the bla(CTX-M-15)-ISEcp1 element. This element seems to have a preferred insertion site at the tnpA gene of a bla(TEM)-carrying Tn3-like transposon, the latter itself being inserted by a transposition event. The IS26-composite transposon, which contains the bla(OXA-1), aac6'-lb-cr and catB4 genes, was inserted into plasmids pEC_L8 and pEC_L46 by homologous recombination rather than a transposition event. Results obtained for pEC_L46 indicated that IS26 also plays an important role in structural rearrangements of the plasmid backbone and seems to facilitate the mobilisation of fragments from other plasmids. CONCLUSIONS: Collectively, these data suggests that IS26 together with ISEcp1 could play a critical role in the evolution of diverse multiresistant plasmids found in clinical Enterobacteriaceae

Antimicrobial resistance (AMR) nanomachines: mechanisms for fluoroquinolone and glycopeptide recognition, efflux and/or deactivation

In this review, we discuss mechanisms of resistance identified in bacterial agents Staphylococcus aureus and the enterococci towards two priority classes of antibiotics—the fluoroquinolones and the glycopeptides. Members of both classes interact with a number of components in the cells of these bacteria, so the cellular targets are also considered. Fluoroquinolone resistance mechanisms include efflux pumps (MepA, NorA, NorB, NorC, MdeA, LmrS or SdrM in S. aureus and EfmA or EfrAB in the enterococci) for removal of fluoroquinolone from the intracellular environment of bacterial cells and/or protection of the gyrase and topoisomerase IV target sites in Enterococcus faecalis by Qnr-like proteins. Expression of efflux systems is regulated by GntR-like (S. aureus NorG), MarR-like (MgrA, MepR) regulators or a two-component signal transduction system (TCS) (S. aureus ArlSR). Resistance to the glycopeptide antibiotic teicoplanin occurs via efflux regulated by the TcaR regulator in S. aureus. Resistance to vancomycin occurs through modification of the D-Ala-D-Ala target in the cell wall peptidoglycan and removal of high affinity precursors, or by target protection via cell wall thickening. Of the six Van resistance types (VanA-E, VanG), the VanA resistance type is considered in this review, including its regulation by the VanSR TCS. We describe the recent application of biophysical approaches such as the hydrodynamic technique of analytical ultracentrifugation and circular dichroism spectroscopy to identify the possible molecular effector of the VanS receptor that activates expression of the Van resistance genes; both approaches demonstrated that vancomycin interacts with VanS, suggesting that vancomycin itself (or vancomycin with an accessory factor) may be an effector of vancomycin resistance. With 16 and 19 proteins or protein complexes involved in fluoroquinolone and glycopeptide resistances, respectively, and the complexities of bacterial sensing mechanisms that trigger and regulate a wide variety of possible resistance mechanisms, we propose that these antimicrobial resistance mechanisms might be considered complex ‘nanomachines’ that drive survival of bacterial cells in antibiotic environments

Remarques concernant la céramique peinte du IIe Âge du Fer en pays éduen

Il faut comprendre dans les termes de « céramique peinte » toute poterie pour laquelle, dans un but artistique, on a fait usage d’une couleur sur le fond écru du récipient autrement que sous forme d’engobe. Dès la phase finale du Ier Age du Fer, la céramique peinte, dont l’origine est encore plus ancienne, devient d’un usage relativement fréquent. Selon certaines modes ou influences diverses, elle évolue. Dans un sens général, d’importants documents ont été livrés par les sépultures de la Mar..

Radiothérapie adaptative en routine ? État de l’art : point de vue du physicien médical

National audienceThe development of both image-guided and intensity-modulated radiotherapy has underlined the question of treatment adaptation to anatomical and/or biological changes occurring during radiotherapy course and modifying delivered dose to the patient. Adaptive radiotherapy has been introduced when several plans are used to treat a patient during radiotherapy. Adaptation may be performed online, offline or in a hybrid way. New images of the patient are needed for adaptive radiotherapy to perform many processes: image registration, segmentation and evaluation of cumulative dose. Deformable image registration methods are generally used to image registration and contours propagation. Fraction and cumulative dose evaluations use deformable image registration methods or more complex methods based on Monte-Carlo calculation. These methods have uncertainties and have to be evaluated. However, evaluation and validation tools are still being developed. The physicist's mission is to ensure that every new technology, such as adaptive radiotherapy, is deployed with highest safety, by technical validation and by implementing a specific quality assurance program. Adaptive radiotherapy implementation still raises many questions, so its potential clinical application requires great caution and should be carefully explored in prospective clinical trials

Calcul de dose à partir d’images Cone Beam CT : état de l’art

National audienceIn external beam radiotherapy, the dose planning is currently based on computed tomography (CT) images. A relation between Hounsfield numbers and electron densities (or mass densities) is necessary for dose calculation taking heterogeneities into account. In image-guided radiotherapy process, the cone beam CT is classically used for tissue visualization and registration. Cone beam CT for dose calculation is also attractive in dose reporting/monitoring perspectives and particularly in a context of dose-guided adaptive radiotherapy. The accuracy of cone beam CT-based dose calculation is limited by image characteristics such as quality, Hounsfield numbers consistency and restrictive sizes of volume acquisition. The analysis of the literature identifies three kinds of strategies for cone beam CT-based dose calculation: establishment of Hounsfield numbers versus densities curves, density override to regions of interest, and deformable registration between CT and cone beam CT images. Literature results show that discrepancies between the reference CT-based dose calculation and the cone beam CT-based dose calculation are often lower than 3%, regardless of the method. However, they can also reach 10% with unsuitable method. Even if the accuracy of the cone beam CT-based dose calculation is independent of the method, some strategies are promising but need improvements in the automating process for a routine implementation

CBCT dose calculation in head and neck adaptive radiotherapy accuracy assessment of four methods

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Planification à partir d’imagerie par résonance magnétique en radiothérapie [MRI-based radiotherapy planning]

National audienceMRI-based radiotherapy planning is a topical subject due to the introduction of a new generation of treatment machines combining a linear accelerator and a MRI. One of the issues for introducing MRI in this task is the lack of information to provide tissue density information required for dose calculation. To cope with this issue, two strategies may be distinguished from the literature. Either a synthetic CT scan is generated from the MRI to plan the dose, or a dose is generated from the MRI based on physical underpinnings. Within the first group, three approaches appear: bulk density mapping assign a homogeneous density to different volumes of interest manually defined on a patient MRI; machine learning-based approaches model local relationship between CT and MRI image intensities from multiple data, then applying the model to a new MRI; atlas-based approaches use a co-registered training data set (CT-MRI) which are registered to a new MRI to create a pseudo CT from spatial correspondences in a final fusion step. Within the second group, physics-based approaches aim at computing the dose directly from the hydrogen contained within the tissues, quantified by MRI. Excepting the physics approach, all these methods generate a synthetic CT called "pseudo CT", on which radiotherapy planning will be finally realized. This literature review shows that atlas- and machine learning-based approaches appear more accurate dosimetrically. Bulk density approaches are not appropriate for bone localization. The fastest methods are machine learning and the slowest are atlas-based approaches. The less automatized are bulk density assignation methods. The physical approaches appear very promising methods. Finally, the validation of these methods is crucial for a clinical practice, in particular in the perspective of adaptive radiotherapy delivered by a linear accelerator combined with an MRI scanner

Radiotherapy for T3b prostate cancer (with seminal involvement) clinical analyses

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