Complete Genome Sequence of a Multidrug-Resistant <i>Klebsiella pneumoniae</i> Environmental Isolate from Zanzibar, Tanzania, Harboring Novel Insertion Elements and Two <i>bla</i>CTX-M-15 Genes

Here, we report the annotated whole-genome sequence of Klebsiella pneumoniae strain KP_3b, isolated in Zanzibar, Tanzania, from plastic litter. The strain is extended-spectrum β-lactamase (ESBL) producing and multidrug resistant, encoding 17 resistance genes, most of which are located on a 230,544-bp plasmid. The isolate contains two copies of the bla(CTX-M-15) gene and novel insertion elements

A shallow water type model to describe the dynamic of thin partially wetting films

Liquid films moving onto a solid substrate are present in a lot of natural and industrial processes and have been the object of a lot of research studies for several decades. In the context of deicing, when a thermal protection system is activated, the supercooled water droplets impacting an aircraft surface don't freeze instantaneously and can coalesce and form a thin liquid film as a result of aerodynamic forces. Experimental studies show that this liquid film isn't always stable and can split into rivulets that may refreeze on unprotected surfaces. The modeling of rivulet flows and thus the accurate prediction of wet and dry surfaces is still an unsolved problem. The objective of this work is to model the motion and the instabilities of partially wetting thin liquid films to derive models for the formation of wet and dry surfaces. This has a direct influence on the estimation of the wall heat and mass fluxes such as evaporation or exchanges with the boundary layer. For thin films, capillary forces generally play an important role and could strongly influence both the motion of the contact line and the development of longitudinal (surface waves) and transversal instabilities (dewetting and rivulets formation). To predict such flows, shallow water models are generally preferred to the full Navier-Stokes equations. The derivation of such models is based on closure assumptions on the normal film velocity profile which can be justified either by asymptotic analysis or by empirical arguments. The main idea of the work consists in reformulating the shallow water equations by introducing a ""disjoining pressure"" to model the effects of a partial wetting. This new term appears like a regularization of the discontinuous forces at the contact line. Emphasis is put on the numerical treatment of the capillary forces, especially those acting in the vicinity of the contact line, since they can strongly influence the development of instabilities. Based on the work of Noble & Vila, we use an augmented conservative system that consists in reducing the order of the shallow water system by adding one evolution equation. This model is suited for numerical purposes since the surface tension term only involves second order derivatives instead of third order derivatives in the classical shallow water systems with two equations. A conservative formulation of the system and the associated energy are derived. One-dimensional numerical simulations using a first order implicit finite volume scheme have been performed. Droplet's stationnary shape, spreading length and time on an horizontal substrate is well recovered for all contact angle. Moreover, based on a linear stability analysis, unstable dewetting regimes of an infinite film of uniform thickness are identified and simulated. The add of a dynamic contact angle, the hysteresis effects and the 3D extension of the method are also in progress

The Role of ISCR1-Borne POUT Promoters in the Expression of Antibiotic Resistance Genes

The ISCR1 (Insertion sequence Common Region) element is the most widespread member of the ISCR family, and is frequently present within γ-proteobacteria that occur in clinical settings. ISCR1 is always associated with the 3′Conserved Segment (3′CS) of class 1 integrons. ISCR1 contains outward-oriented promoters POUT, that may contribute to the expression of downstream genes. In ISCR1, there are two POUT promoters named PCR1-1 and PCR1-2. We performed an in silico analysis of all publically available ISCR1 sequences and identified numerous downstream genes that mainly encode antibiotic resistance genes and that are oriented in the same direction as the POUT promoters. Here, we showed that both PCR1-1 and PCR1-2 significantly increase the expression of the downstream genes blaCTX-M-9 and dfrA19. Our data highlight the role of ISCR1 in the expression of antibiotic resistance genes, which may explain why ISCR1 is so frequent in clinical settings

Characterization of bacterial insertion sequences ISCR involved in antibiotic resistance

Les ISCR constituent une famille de séquences d’insertions bactériennes décrits récemment dans des contextes cliniques et d’antibiorésistance. Les transposases codées par ces IS appartiennent à la famille des HUH transposases qui transposent selon un mécanisme en cercle roulant. Néanmoins, aucune donnée expérimentale n’existe à ce jour. La famille ISCR compte 19 membres mais ont été peu caractérisés. C’est pourquoi, nous avons fait une mise à jour des informations sur ces éléments en analysant in silico les principales caractéristiques par une étude in silico. Nous nous sommes ensuite intéressés à l’implication de l’élément ISCR1 dans l’expression de la région variable en aval. Cet élément contient deux promoteurs orientés vers l’extérieur (POUT) dans sa région en 3’. Après une analyse de la diversité des gènes, nous avons remarqué que la plupart des gènes en aval étaient orientés dans le même sens que ces POUT et qu’ils pouvaient être exprimés à partir des deux promoteurs. Nous avons montré que pour deux gènes de résistance dfrA19 et blaCTX-M-9, ces promoteurs augmentent le niveau d’expression. De plus, la région contenant les deux promoteurs est nécessaire pour que l’expression de blaCTX-M-9 confère un phénotype de résistance. En parallèle, nous avons déterminé la régulation du promoteur du gène de la transposase de ISCR1. Nous avons identifié des motifs de régulation pour les régulateurs LexA et OmpR et déterminé expérimentalement que le promoteur du gène de la transposase de ISCR1 était régulé de façon négative par la protéine LexA, régulateur majeur de la réponse SOS et de façon positive par la protéine OmpR en conditions hypo-osmotiques. Nous proposons donc un modèle selon lequel ISCR1 est un élément dont la mobilité serait conditionnée par des facteurs environnementaux et en même temps, assurerait l’expression constitutive de gènes en aval, notamment impliqués dans l’antibiorésistance.ISCR are a bacterial insertion sequences, recently described in clinical settings, frequently related to antibiotic resistance. These ISCR-encoded transposases belong to the well-known HUH transposases family, which transpose by rolling-circle replication. However, the transposition mechanism of ISCR transposases has not been shown experimentally. ISCR family includes 19 members and has not been well characterized yet. Therefore, we updated in silico already known characteristics for each ISCR element. Then, we investigated the involvement of ISCR1 in the expression of the downstream genes. Indeed, ISCR1 carries two outward-oriented promoters called POUT. By analyzing the diversity of the downstream region, we found that most of genes were in the same orientation as POUT promoters, suggesting these downstream genes are expressed from POUT. It thus showed that these two promoters are able to express two antibiotic resistance genes (dfrA19 and blaCTX-M-9 ). Moreover, the region containing POUT is essential to provide an ESBL-resistance phenotype for blaCTX-M-9 gene. Moreover, we also wanted to analyze the regulatory network involved in the expression of the ISCR1 transposase, RCR1. We experimentally determined that two regulatory proteins LexA and OmpR, involved in response to different stress (DNA damages and osmotic shock), control the activity of rcr1 promoter. LexA protein represses Prcr1 whereas OmpR activates Prcr1 in hypo-osmotic conditions. Here, we propose a model in which ISCR1 transposition would be the control of environmental stresses and at the same time, insured the expression of downstream antibiotic resistance genes

Caractérisation des séquences d'insertions ISCR bactériennes impliquées dans la résistance aux antibiotiques

ISCR are a bacterial insertion sequences, recently described in clinical settings, frequently related to antibiotic resistance. These ISCR-encoded transposases belong to the well-known HUH transposases family, which transpose by rolling-circle replication. However, the transposition mechanism of ISCR transposases has not been shown experimentally. ISCR family includes 19 members and has not been well characterized yet. Therefore, we updated in silico already known characteristics for each ISCR element. Then, we investigated the involvement of ISCR1 in the expression of the downstream genes. Indeed, ISCR1 carries two outward-oriented promoters called POUT. By analyzing the diversity of the downstream region, we found that most of genes were in the same orientation as POUT promoters, suggesting these downstream genes are expressed from POUT. It thus showed that these two promoters are able to express two antibiotic resistance genes (dfrA19 and blaCTX-M-9 ). Moreover, the region containing POUT is essential to provide an ESBL-resistance phenotype for blaCTX-M-9 gene. Moreover, we also wanted to analyze the regulatory network involved in the expression of the ISCR1 transposase, RCR1. We experimentally determined that two regulatory proteins LexA and OmpR, involved in response to different stress (DNA damages and osmotic shock), control the activity of rcr1 promoter. LexA protein represses Prcr1 whereas OmpR activates Prcr1 in hypo-osmotic conditions. Here, we propose a model in which ISCR1 transposition would be the control of environmental stresses and at the same time, insured the expression of downstream antibiotic resistance genes.Les ISCR constituent une famille de séquences d’insertions bactériennes décrits récemment dans des contextes cliniques et d’antibiorésistance. Les transposases codées par ces IS appartiennent à la famille des HUH transposases qui transposent selon un mécanisme en cercle roulant. Néanmoins, aucune donnée expérimentale n’existe à ce jour. La famille ISCR compte 19 membres mais ont été peu caractérisés. C’est pourquoi, nous avons fait une mise à jour des informations sur ces éléments en analysant in silico les principales caractéristiques par une étude in silico. Nous nous sommes ensuite intéressés à l’implication de l’élément ISCR1 dans l’expression de la région variable en aval. Cet élément contient deux promoteurs orientés vers l’extérieur (POUT) dans sa région en 3’. Après une analyse de la diversité des gènes, nous avons remarqué que la plupart des gènes en aval étaient orientés dans le même sens que ces POUT et qu’ils pouvaient être exprimés à partir des deux promoteurs. Nous avons montré que pour deux gènes de résistance dfrA19 et blaCTX-M-9, ces promoteurs augmentent le niveau d’expression. De plus, la région contenant les deux promoteurs est nécessaire pour que l’expression de blaCTX-M-9 confère un phénotype de résistance. En parallèle, nous avons déterminé la régulation du promoteur du gène de la transposase de ISCR1. Nous avons identifié des motifs de régulation pour les régulateurs LexA et OmpR et déterminé expérimentalement que le promoteur du gène de la transposase de ISCR1 était régulé de façon négative par la protéine LexA, régulateur majeur de la réponse SOS et de façon positive par la protéine OmpR en conditions hypo-osmotiques. Nous proposons donc un modèle selon lequel ISCR1 est un élément dont la mobilité serait conditionnée par des facteurs environnementaux et en même temps, assurerait l’expression constitutive de gènes en aval, notamment impliqués dans l’antibiorésistance

An experimental study on gravity-driven film: influence of viscosity, wall surface treatment and transitory state

International audienceThe formation of water rivulet on airplane wings modifies the momentum and heat transfers between the liquid, the wall and the air. In icing conditions, this phenomenon impacts the shape of the ice formed and thus the level of the risk induced. Previous published work on rivulets mostly concerns high wetting and viscous fluids. In order to understand the influence of substrate wettability and fluid viscosity on wall liquid flow, a gravity driven wall film is studied for different liquid and wall conditions. After a transitory step, an equilibrium state is reached. Two steady flow configurations are observed: the converging film equilibrium state and the constant-width equilibrium film. During the transitory step, rivulets appear. Their behavior influences the final equilibrium state through the interaction between the liquid and the wall. So, their contact lines can either recede during the transitory state or stay pinned on the equilibrium state. Mechanisms involving a modification of the contact angle hysteresis are suspected of playing a part in the discrimination between the two equilibrium configurations. Observation shows that, as the fluid viscosity increases, the film is wider. Simulated films behavior agrees with experimental results for the receded contact lines equilibrium but fails to reproduce the other state formation

Landscape of Resistance-Nodulation-Cell Division (RND)-Type Efflux Pumps in Enterobacter cloacae Complex

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Prolonged inflammatory gene response following soman-induced seizures in mice.

International audienceFollowing exposure to the organophosphorus nerve agent soman, the development of long-lasting seizures and build-up of irreversible seizure-related brain damage (SRBD) still represent a therapeutic challenge. A neuro-inflammatory reaction takes place in the brain after poisoning but its characteristics and potential role in SRBD and post-status epilepticus epileptogenesis is not well understood. In the present study we have analyzed by quantitative RT-PCR the time course of changes in mRNA levels of IL-1beta, TNFalpha, IL-6, ICAM-1 and SOCS3 in hippocampus, whole cortex and cerebellum in a mouse model of severe seizures and neuropathy up to 7 days after poisoning. Mice received an injection of the oxime HI-6 (50mg/kg) 5 min prior to the administration of a convulsive dose of soman (172 microg/kg). An important and highly significant increase of the five mRNA levels was recorded in cortex and hippocampus. In the cortex, the activation was generally detected as early as 1h post-intoxication with a peak response recorded between 6 and 24h. In the hippocampus, the gene up-regulation was delayed to 6h post-soman and the peak response observed between 24 and 48 h. After peaking, the response declined (except for ICAM in the hippocampus) but remained elevated, some of them significantly, at day 7. Interestingly, in the cerebellum, some changes were also observed but were several fold smaller. In conclusion, the present study indicates a quick neuro-inflammatory gene response that does not subside over 7 days suggesting a potential role in the neurological consequences of soman-induced status epilepticus

Pallidal stimulation in advanced Parkinson's patients with contraindications for subthalamic stimulation.

International audienceThe aim of this study was to evaluate the efficacy and safety of bilateral pallidal (GPi) deep brain stimulation (DBS) 6 months after surgery in advanced parkinsonian patients whose dopa-resistant axial motor signs or cognitive decline constituted contraindications for subthalamic nucleus (STN) DBS. Seventeen patients with a mean age of 59.3 ± 7.1 years (range, 45-70), mean disease duration of 12.5 ± 4.3 years (range, 7-20), and contraindications for STN DBS, underwent bilateral GPi DBS. They were evaluated before surgery and 6 months afterward, in accordance with Core Assessment Program for Intracerebral Transplantation recommendations. There were mean improvements of 41.1% in the UPDRS III motor score in the off-dopa condition and 20.3% in the activities of daily living score. Motor fluctuations were reduced by 22.9% and dyskinesias by 68.6%. Axial motor signs improved in the off-dopa condition by 34.2%. Neuropsychological performances remained unchanged at the 6-month assessment. Bilateral GPi DBS is both safe and effective in advanced parkinsonian patients with untreatable motor fluctuations, for whom STN DBS is contraindicated due to dopa-resistant axial motor signs or cognitive decline. As such, it should be regarded as a viable option for these patients