Engineered single nucleotide polymorphisms in the mosquito MEK docking site alter Plasmodium berghei development in Anopheles gambiae.

BackgroundSusceptibility to Plasmodium infection in Anopheles gambiae has been proposed to result from naturally occurring polymorphisms that alter the strength of endogenous innate defenses. Despite the fact that some of these mutations are known to introduce non-synonymous substitutions in coding sequences, these mutations have largely been used to rationalize knockdown of associated target proteins to query the effects on parasite development in the mosquito host. Here, we assay the effects of engineered mutations on an immune signaling protein target that is known to control parasite sporogonic development. By this proof-of-principle work, we have established that naturally occurring mutations can be queried for their effects on mosquito protein function and on parasite development and that this important signaling pathway can be genetically manipulated to enhance mosquito resistance.MethodsWe introduced SNPs into the A. gambiae MAPK kinase MEK to alter key residues in the N-terminal docking site (D-site), thus interfering with its ability to interact with the downstream kinase target ERK. ERK phosphorylation levels in vitro and in vivo were evaluated to confirm the effects of MEK D-site mutations. In addition, overexpression of various MEK D-site alleles was used to assess P. berghei infection in A. gambiae.ResultsThe MEK D-site contains conserved lysine residues predicted to mediate protein-protein interaction with ERK. As anticipated, each of the D-site mutations (K3M, K6M) suppressed ERK phosphorylation and this inhibition was significant when both mutations were present. Tissue-targeted overexpression of alleles encoding MEK D-site polymorphisms resulted in reduced ERK phosphorylation in the midgut of A. gambiae. Furthermore, as expected, inhibition of MEK-ERK signaling due to D-site mutations resulted in reduction in P. berghei development relative to infection in the presence of overexpressed catalytically active MEK.ConclusionMEK-ERK signaling in A. gambiae, as in model organisms and humans, depends on the integrity of conserved key residues within the MEK D-site. Disruption of signal transmission via engineered SNPs provides a purposeful proof-of-principle model for the study of naturally occurring mutations that may be associated with mosquito resistance to parasite infection as well as an alternative genetic basis for manipulation of this important immune signaling pathway

Development of a relationship between external measurements and reinforcement stress

As many countries around the world face an aging infrastructure crisis, there is an increasing need to develop more accurate monitoring and assessment techniques for reinforced concrete structures. One of the challenges associated with assessing existing infrastructure is correlating externally measured parameters such as crack widths and surface strains with reinforcement stresses as this is dependent on a number of variables. The current research investigates how the use of distributed fiber optic sensors to measure reinforcement strain can be correlated with digital image correlation measurements of crack widths to relate external crack width measurements to reinforcement stresses. An initial set of experiments was undertaken involving a series of small-scale beam specimens tested in three-point bending with variable reinforcement properties. Relationships between crack widths and internal reinforcement strains were observed including that both the diameter and number of bars affected the measured maximum strain and crack width. A model that uses measured crack width to estimate reinforcement strain was presented and compared to the experimental results. The model was found to provide accurate estimates of load carrying capacity for a given crack width, however, the model was potentially less accurate when crack widths were used to estimate the experimental reinforcement strains. The need for more experimental data to validate the conclusions of this research was also highlighted.The authors would like to thank the Natural Science and Engineering Research Council of Canada, the Canada Foundation for Innovation, and the Ontario Research Fund for their financial support of this research. The authors would also like to thank Adam Hoag, Jaime Escobar, Neil Porter, and Paul Thrasher for their assistance with the experimental program.This is the author accepted manuscript. The final version is available from SPIE at http://proceedings.spiedigitallibrary.org/proceeding.aspx?articleid=2213474

Impact of Tumor-Derived CCL2 on Macrophage Effector Function

Monocyte chemoattractant protein-1 (MCP-1, CCL2) is produced by many different types of cells. In the current investigation, the effect of tumor-derived CCL2 on macrophages was evaluated to determine the extent to which this chemokine influenced the innate immune response to cancer. To do this, we used the 4T1 murine mammary carcinoma cell line that constitutively expresses CCL2 and generated 4T1 expressing an antisense CCL2 transcript. The antisense-CCL2-expressing 4T1 produced no detectable CCL2. Macrophages from female BALB/c mice were exposed to supernatants from these tumor cells. The results showed that tumor-derived CCL2 was capable of modulating cytokine gene expression but not protein production in resting, activated, and tumor-associated macrophages. In addition, tumor-derived CCL2 did not affect phagocytic activity, nitric oxide production, or cytolytic activity of the macrophages. Overall, these data suggest that tumor-derived CCL2 does not directly influence macrophage-mediated antitumor activity

The Motion for New Trial and Its Constitutional Tension

Either the judge or the jury must decide facts and, to the extent that we take this responsibility, we lessen the jury function. Our duty to preserve this one of the Bill of Rights may be peculiarly difficult, for here it is our own power which we must restrain.

Introduction à la gouvernance clinique : historique, composantes et conceptualisation renouvelée pour l’amélioration de la qualité et de la performance des organisations de santé

La recherche de l’efficience et de l’excellence dans le domaine de la santé amène organisations et professionnels à travailler conjointement à l’amélioration des processus cliniques. La gouvernance clinique se veut un lieu d’action collective, de mobilisation de relations et de savoirs entre les acteurs impliqués dans l’organisation et la dispensation des soins et des services de santé. Elle est un espace où s’exerce l’autorité des différents acteurs en vue d’améliorer la qualité des soins et des services de santé. Elle vise à rapprocher la perspective organisationnelle et la perspective professionnelle par le développement et l’implantation d’initiatives visant les meilleures pratiques cliniques et organisationnelles.Pour améliorer la qualité des soins, Ferlie et al(MilbankQ2001.79:281)[1] suggèrent de créer un alignement entre les différents niveaux de soins: l’individu, l’équipe, l’organisation et le système. Les principes de la gouvernance clinique tentent de produire cette synergie en impliquant les professionnels dans le renouvellement de l’organisation et enimpliquant l’organisation dans l’instrumentation et la régulation des pratiques des professionnels en agissant aux différents niveaux de soins. Notre objectif est de présenter les origines du concept de gouvernance clinique, de décrire ses composantes et de proposer des assises théoriques afin demieux comprendre les dynamiquesde changement associées à la gouvernance clinique et faciliter son implantation. Nous exposons le courant britannique fondateur puis la notion de gouvernance développée par HatchuelA (Paris: ÉAube ; 2000) [2] et présentons les caractéristiques de l’organisation pouvant soutenir l’engagement des professionnels envers les principes de qualité. Enfin, nous proposons une modélisation renouvelée de la gouvernance clinique permettant de concilier l’organisationnel et la clinique, et de potentialiser les capacités d’action de chacun

First evidence for a gravitational lensing-induced echo in gamma rays with Fermi LAT

Aims. This article shows the first evidence for gravitational lensing phenomena in high energy gamma-rays. This evidence comes from the observation of a gravitational lens induced echo in the light curve of the distant blazar PKS 1830-211. Methods. Traditional methods for the estimation of time delays in gravitational lensing systems rely on the cross-correlation of the light curves of the individual images. In this paper, we use 300 MeV-30 GeV photons detected by the Fermi-LAT instrument. The Fermi-LAT instrument cannot separate the images of known lenses. The observed light curve is thus the superposition of individual image light curves. The Fermi-LAT instrument has the advantage of providing long, evenly spaced, time series. In addition, the photon noise level is very low. This allows to use directly Fourier transform methods. Results. A time delay between the two compact images of PKS 1830-211 has been searched for both by the autocorrelation method and the "double power spectrum" method. The double power spectrum shows a 3 {\sigma} evidence for a time delay of 27.5 $\pm$ 1.3 days, consistent with the result from Lovell et al. (1998). The relative uncertainty on the time delay estimation is reduced from 20% to 5%.Comment: submitted to A&

On the solar nickel and oxygen abundances

Determinations of the solar oxygen content relying on the neutral forbidden transition at 630 nm depend upon the nickel abundance, due to a Ni I blend. Here we rederive the solar nickel abundance, using the same ab initio 3D hydrodynamic model of the solar photosphere employed in the recent revision of the abundances of C, N, O and other elements. Using 17 weak, unblended lines of Ni I together with the most accurate atomic and observational data available we find log epsilon_Ni = 6.17 +/- 0.02 (statistical) +/- 0.05 (systematic), a downwards shift of 0.06 to 0.08 dex relative to previous 1D-based abundances. We investigate the implications of the new nickel abundance for studies of the solar oxygen abundance based on the [O I] 630 nm line in the quiet Sun. Furthermore, we demonstrate that the oxygen abundance implied by the recent sunspot spectropolarimetric study of Centeno & Socas-Navarro needs to be revised downwards from log epsilon_O = 8.86 +/- 0.07 to 8.71 +/- 0.10. This revision is based on the new nickel abundance, application of the best available gf-value for the 630 nm forbidden oxygen line, and a more transparent treatment of CO formation. Determinations of the solar oxygen content relying on forbidden lines now appear to converge around log epsilon_O = 8.7.Comment: v2 matches published versio