BACKGROUND AND PURPOSE The cholinergic agonist levamisole is widely used to treat parasitic nematode infestations. This anthelmintic drug paralyses worms by activating a class of levamisole‐sensitive ...acetylcholine receptors (L‐AChRs) expressed in nematode muscle cells. However, levamisole efficacy has been compromised by the emergence of drug‐resistant parasites, especially in gastrointestinal nematodes such as Haemonchus contortus. We report here the first functional reconstitution and pharmacological characterization of H. contortus L‐AChRs in a heterologous expression system.
EXPERIMENTAL APPROACH In the free‐living nematode Caenorhabditis elegans, five AChR subunit and three ancillary protein genes are necessary in vivo and in vitro to synthesize L‐AChRs. We have cloned the H. contortus orthologues of these genes and expressed them in Xenopus oocytes. We reconstituted two types of H. contortus L‐AChRs with distinct pharmacologies by combining different receptor subunits.
KEY RESULTS The Hco‐ACR‐8 subunit plays a pivotal role in selective sensitivity to levamisole. As observed with C. elegans L‐AChRs, expression of H. contortus receptors requires the ancillary proteins Hco‐RIC‐3, Hco‐UNC‐50 and Hco‐UNC‐74. Using this experimental system, we demonstrated that a truncated Hco‐UNC‐63 L‐AChR subunit, which was specifically detected in a levamisole‐resistant H. contortus isolate, but not in levamisole‐sensitive strains, hampers the normal function of L‐AChRs, when co‐expressed with its full‐length counterpart.
CONCLUSIONS AND IMPLICATIONS We provide the first functional evidence for a putative molecular mechanism involved in levamisole resistance in any parasitic nematode. This expression system will provide a means to analyse molecular polymorphisms associated with drug resistance at the electrophysiological level.
Measurements of midrapidity charged-particle multiplicity distributions, dNch/dη, and midrapidity transverse-energy distributions, dET/dη, are presented for a variety of collision systems and ...energies. Included are distributions for Au+Au collisions at sNN=200, 130, 62.4, 39, 27, 19.6, 14.5, and 7.7 GeV, Cu+Cu collisions at sNN=200 and 62.4 GeV, Cu+Au collisions at sNN=200 GeV, U+U collisions at sNN=193 GeV, d+Au collisions at sNN=200 GeV, He3+Au collisions at sNN=200 GeV, and p+p collisions at sNN=200 GeV. Centrality-dependent distributions at midrapidity are presented in terms of the number of nucleon participants, Npart, and the number of constituent quark participants, Nqp. For all A+A collisions down to sNN=7.7 GeV, it is observed that the midrapidity data are better described by scaling with Nqp than scaling with Npart. Also presented are estimates of the Bjorken energy density, BJ, and the ratio of dET/dη to dNch/dη, the latter of which is seen to be constant as a function of centrality for all systems.
Isotopic effects in the fragmentation of excited target residues following collisions of 12C on (112,124)Sn at incident energies of 300 and 600 MeV per nucleon were studied with the INDRA 4pi ...detector. The measured yield ratios for light particles and fragments with atomic number Z < or = 5 obey the exponential law of isotopic scaling. The deduced scaling parameters decrease strongly with increasing centrality to values smaller than 50% of those obtained for the peripheral event groups. Symmetry-term coefficients, deduced from these data within the statistical description of isotopic scaling, are near gamma = 25 MeV for peripheral and gamma < 15 MeV for central collisions.
Azimuthal angle (Delta phi) correlations are presented for a broad range of transverse momentum (0.4 < p(T) < 10 GeV/c) and centrality (0-92%) selections for charged hadrons from dijets in Au+Au ...collisions at root s(NN) = 200 GeV. With increasing p(T), the away-side Delta phi distribution evolves from a broad and relatively flat shape to a concave shape, then to a convex shape. Comparisons with p + p data suggest that the away-side distribution can be divided into a partially suppressed "head" region centered at Delta phi similar to pi, and an enhanced "shoulder" region centered at Delta phi similar to pi +/- 1.1. The p(T) spectrum for the associated hadrons in the head region softens toward central collisions. The spectral slope for the shoulder region is independent of centrality and trigger p(T). The properties of the near-side distributions are also modified relative to those in p + p collisions, reflected by the broadening of the jet shape in Delta phi and Delta eta, and an enhancement of the per-trigger yield. However, these modifications seem to be limited to p(T)less than or similar to 4 GeV/c, above which both the hadron pair shape and per-trigger yield become similar to p + p collisions. These observations suggest that both the away- and near-side distributions contain a jet fragmentation component which dominates for p(T) greater than or similar to 5 GeV/c and a medium-induced component which is important for p(T) less than or similar to 4 GeV/c. We also quantify the role of jets at intermediate and low p(T) through the yield of jet-induced pairs in comparison with binary scaled p + p pair yield. The yield of jet-induced pairs is suppressed at high pair proxy energy (sum of the p(T) magnitudes of the two hadrons) and is enhanced at low pair proxy energy. The former is consistent with jet quenching; the latter is consistent with the enhancement of soft hadron pairs due to transport of lost energy to lower p(T).
Methotrexate is the first line of treatment of rheumatoid arthritis. Since many patients become unresponsive to methotrexate treatment, only very expensive biological therapies are effective and ...increased methotrexate tolerance strategies need to be identified. Here we propose the encapsulation of methotrexate in a new liposomal formulation using a hydrophobic fragment of surfactant protein conjugated to a linker and folate to enhance their tolerance and efficacy. In this study we aim to evaluate the efficiency of this system to treat rheumatoid arthritis, by targeting folate receptor β present at the surface of activated macrophages, key effector cells in this pathology. The specificity of our liposomal formulation to target folate receptor β was investigated both in vitro as in vivo using a mouse model of arthritis (collagen-induced arthritis in DBA/1J mice strain). In both systems, the liposomal constructs were shown to be highly specific and efficient in targeting folate receptor β. These liposomal formulations also significantly increase the clinical benefit of the encapsulated methotrexate in vivo in arthritic mice, together with reduced expression of CD39 and CD73 ectonucleotidases by joint-infiltrating macrophages. Thus, our formulation might be a promising cost effective way to treat rheumatoid arthritis and delay or reduce methotrexate intolerance.
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► A new method to analyze material outgassing is presented. ► From the experiment the ratio and nature of gases is detected. ► The knowledge of material outgassing properties is a key ...step for vacuum thin film packaging.
In this paper, a new method to characterize materials outgassing behavior is detailed. The understanding of these phenomena during all the process steps is required to improve vacuum thin film packaging. Starting from a standard hermetic fabrication process, each material of each layer is analyzed separately thanks to this innovative experimental method coupled with mass. Samples are sealed in an ampoule and a direct measure of pressure vs time and temperature is performed. From the variation of the pressure, the critical temperature (defined as the temperature for which pressure is above a given threshold) and the kinetics of the phenomenon are evaluated. The results are compared to thermodynamic simulations performed with the FactSage software. A full study of CVD deposited SiO2 is presented as an example. The results show all the potential of this experimental method and the improvements that have to be done to be able later onto choose best materials for thin film vacuum packaging.
Reaction mechanism analyses performed with a 4pi detector for the systems 208Pb + Ge, 238U + Ni and 238U + Ge, combined with analyses of the associated reaction time distributions, provide us with ...evidence for nuclei with Z=120 and 124 living longer than 10(-18) s and arising from highly excited compound nuclei. By contrast, the neutron deficient nuclei with Z=114 possibly formed in 208Pb + Ge reactions have shorter lifetimes, close to or below the sensitivity limit of the experiment.
In this paper, we present the first measurement of elliptic (v2) and triangular (v3) flow in high-multiplicity 3He+Aucollisions at √sNN=200 GeV. Two-particle correlations, where the particles have a ...large separation in pseudorapidity, are compared in 3He+Au and in p+p collisions and indicate that collective effects dominate the second and third Fourier components for the correlations observed in the 3He+Ausystem. The collective behavior is quantified in terms of elliptic v2 and triangular v3 anisotropy coefficients measured with respect to their corresponding event planes. The v2 values are comparable to those previously measured in d+Au collisions at the same nucleon-nucleon center-of-mass energy. Comparisons with various theoretical predictions are made, including to models where the hot spots created by the impact of the three 3He nucleons on the Au nucleus expand hydrodynamically to generate the triangular flow. Finally, the agreement of these models with data may indicate the formation of low-viscosity quark-gluon plasma even in these small collision systems.
This manuscript analyses the use of newly developed hybrid gadolinium oxide nanoparticles as cell-labeling tracers. The nanoparticles are core-shell particles composed of a core of gadolinium oxide ...of 2-4 nm and a protecting shell of polysiloxane 1-3 nm where different organic dyes (fluoresceine isothiocyanate (FITC) or rhodamine B isothiocyanate (RBITC)) are embedded. They are functionalized with poly(ethylene glycol)bis(carboxymethyl) to ensure their colloidal stability in biological buffers. These particles are potential multi-labeling tracers (magnetic and optical). In this paper, we show by optical imaging that they can be efficiently internalized in cells without cell alteration. The in-vitro uptake of the nanoparticles was followed in two cell lines (human fibroblasts and a human adenocarnima cell lines MCF7 cells). Nanoparticles distribution within cells was analysed by confocal analysis, and gadolinium concentration within cells was quantified by mass spectrometry (ICP-MS analysis). Nanoparticles uptake is found to be fast and efficient for both cell lines, with fluorescent labeling visible after 10 min of incubation whatever the nature of the fluorophore. The fluorescent intensity is mainly found as concentrated dots in the perinuclear region of the cells and decreases with the number of days in culture, but is still easily detectable after 3 days in culture. No significant effect on cell growth was detected. Finally, we show in this study the protective effect of the polysiloxane layer: encapsulation of RBITC within the polysiloxane shell, leads to a better photostability of this low cost dye than Cy3 and even reach a level comparable to Alexa 595. With their high photostability and long-lasting contrast properties, these hybrid luminescent nanoparticles appears thus as a versatile solution to assess multiple cell fate both in in-vitro cell model as well as in-vivo.
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