La performance : un lieu d'échanges et de controverses

peer reviewe

Aminopeptidase B, a glucagon-processing enzyme: site directed mutagenesis of the Zn2+-binding motif and molecular modelling

<p>Abstract</p> <p>Background</p> <p>Aminopeptidase B (Ap-B; EC 3.4.11.6) catalyzes the cleavage of basic residues at the N-terminus of peptides and processes glucagon into miniglucagon. The enzyme exhibits, <it>in vitro</it>, a residual ability to hydrolyze leukotriene A₄into the pro-inflammatory lipid mediator leukotriene B₄. The potential bi-functional nature of Ap-B is supported by close structural relationships with LTA₄hydrolase (LTA₄H ; EC 3.3.2.6). A structure-function analysis is necessary for the detailed understanding of the enzymatic mechanisms of Ap-B and to design inhibitors, which could be used to determine the complete <it>in vivo </it>functions of the enzyme.</p> <p>Results</p> <p>The rat Ap-B cDNA was expressed in <it>E. coli </it>and the purified recombinant enzyme was characterized. 18 mutants of the H³²⁵EXXHX₁₈E³⁴⁸Zn²⁺-binding motif were constructed and expressed. All mutations were found to abolish the aminopeptidase activity. A multiple alignment of 500 sequences of the M1 family of aminopeptidases was performed to identify 3 sub-families of exopeptidases and to build a structural model of Ap-B using the x-ray structure of LTA₄H as a template. Although the 3D structures of the two enzymes resemble each other, they differ in certain details. The role that a loop, delimiting the active center of Ap-B, plays in discriminating basic substrates, as well as the function of consensus motifs, such as RNP1 and Armadillo domain are discussed. Examination of electrostatic potentials and hydrophobic patches revealed important differences between Ap-B and LTA₄H and suggests that Ap-B is involved in protein-protein interactions.</p> <p>Conclusion</p> <p>Alignment of the primary structures of the M1 family members clearly demonstrates the existence of different sub-families and highlights crucial residues in the enzymatic activity of the whole family. <it>E. coli </it>recombinant enzyme and Ap-B structural model constitute powerful tools for investigating the importance and possible roles of these conserved residues in Ap-B, LTA₄H and M1 aminopeptidase catalytic sites and to gain new insight into their physiological functions. Analysis of Ap-B structural model indicates that several interactions between Ap-B and proteins can occur and suggests that endopeptidases might form a complex with Ap-B during hormone processing.</p

Serum proteins electrophoresis changes in dogs infected with Dirofilaria immitis, with Dirofi/aria repens and with both of them

L’électrophorèse des protéines sériques sur acétate de cellulose (migration de 4 heures, bande et courbe de même longueur) a été réalisée sur 4 groupes de chiens: l.Non infestés. 2. Infestés par Dirofilaria immitis. 3. Infestés par Dirofilaria repens. 4. Infestation mixte. L’étude statistique des résultats indique une augmentation significative des globulines gamma pour les groupes 2 et 4.Serum proteins electrophoresis on cellulose ace tat (four hours migration, same length for bands and curves) was carried out on four groups of dogs. Group 1 : not infected, group 2 : only infected with Dirofilaria immitis, group 3 : only infected with Dirofilaria repens and group 4 : infected with both of them. A statistic study shows that the serum globulins level rises to a significative value in groups two and four

MICROSCOPE mission analysis, requirements and expected performance

The MICROSCOPE mission aimed to test the Weak Equivalence Principle (WEP) to a precision of $10^{-15}$. The WEP states that two bodies fall at the same rate on a gravitational field independently of their mass or composition. In MICROSCOPE, two masses of different compositions (titanium and platinum alloys) are placed on a quasi-circular trajectory around the Earth. They are the test-masses of a double accelerometer. The measurement of their accelerations is used to extract a potential WEP violation that would occur at a frequency defined by the motion and attitude of the satellite around the Earth. This paper details the major drivers of the mission leading to the specification of the major subsystems (satellite, ground segment, instrument, orbit...). Building upon the measurement equation, we derive the objective of the test in statistical and systematic error allocation and provide the mission's expected error budget.Comment: References update

In-Person and Remote Workshops for People With Neurocognitive Disorders: Recommendations From a Delphi Panel

Workshops using arts and board games are forms of non-pharmacological intervention widely employed in seniors with neurocognitive disorders. However, clear guidelines on how to conduct these workshops are missing. The objective of the Art and Game project (AGAP) was to draft recommendations on the structure and content of workshops for elderly people with neurocognitive disorders and healthy seniors, with a particular focus on remote/hybrid workshops, in which at least a part of the participants is connected remotely. Recommendations were gathered using a Delphi methodology. The expert panel (N = 18) included experts in the health, art and/or board games domains. They answered questions via two rounds of web-surveys, and then discussed the results in a plenary meeting. Some of the questions were also shared with the general public (N = 101). Both the experts and the general public suggested that organizing workshops in a hybrid format (some face-to-face sessions, some virtual session) is feasible and interesting for people with neurocognitive disorders. We reported guidelines on the overall structure of workshops, practical tips on how to organize remote workshops, and a SWOT analysis of the use of remote/hybrid workshops. The guidelines may be employed by clinicians to decide, based on their needs and constraints, what interventions and what kind of workshop format to employ, as well as by researcher to standardize procedures to assess the effectiveness of non-pharmacological treatments for people with neurocognitive disorders

Family trios analysis of common polymorphisms in the obestatin/ghrelin, BDNF and AGRP genes in patients with Anorexia nervosa: Association with subtype, body-mass index, severity and age of onset.

Anorexia nervosa (AN) affects 0.3% of young girls with a mortality of 6%/decade and is strongly familial with genetic factors. Ghrelin is an upstream regulator of the orexigenic peptides NPY and AgRP and acts as a natural antagonist to leptin's effects on NPY/AgRP-expressing neurons, resulting in an increase in feeding and body weight. Obestatin which counteracts ghrelin action on feeding is derived from the same propeptide than ghrelin. BDNF has been involved in body weight regulation and its Val66Met polymorphism associated with AN. We therefore re-investigated the association between AN and the Leu72Met and Gln90Leu polymorphisms of the prepro-ghrelin/obestatin gene, the Ala67Thr polymorphism of AgRP and the Val66Met polymorphism of BDNF taking into account clinical subtypes (restrictive-ANR-and bingeing/purging-ANB-subtypes). Family trios study of these 4 single nucleotide polymorphisms were performed in 114 probands with AN and both their parents recruited in two specialized French centres. A transmission disequilibrium was observed for the Leu72Met SNP of the preproghrelin gene and for the Ala67Thr SNP of the AgRP gene. When stratified by clinical subtype, these two polymorphisms were preferentially transmitted for the trios with a bingeing/purging proband. An excess of transmission of the Gln90Leu72 preproghrelin/obestatin haplotype in patients with AN was observed. These results do not provide evidence for a preferential transmission of the 66Met allele of BDNF but support the hypothesis that ghrelin and AGRP polymorphisms confers susceptibility to AN. Further simultaneous analysis of genetic variants of the biological determinants of energy metabolism and feeding behaviour in very large populations should contribute to the understanding of the high degree of heritability of eating disorders and to the description of pathophysiological patterns leading to life-threatening conditions in a highly redundant system

Characterization of Reemerging Chikungunya Virus

An unprecedented epidemic of chikungunya virus (CHIKV) infection recently started in countries of the Indian Ocean area, causing an acute and painful syndrome with strong fever, asthenia, skin rash, polyarthritis, and lethal cases of encephalitis. The basis for chikungunya disease and the tropism of CHIKV remain unknown. Here, we describe the replication characteristics of recent clinical CHIKV strains. Human epithelial and endothelial cells, primary fibroblasts and, to a lesser extent, monocyte-derived macrophages, were susceptible to infection and allowed viral production. In contrast, CHIKV did not replicate in lymphoid and monocytoid cell lines, primary lymphocytes and monocytes, or monocyte-derived dendritic cells. CHIKV replication was cytopathic and associated with an induction of apoptosis in infected cells. Chloroquine, bafilomycin-A1, and short hairpin RNAs against dynamin-2 inhibited viral production, indicating that viral entry occurs through pH-dependent endocytosis. CHIKV was highly sensitive to the antiviral activity of type I and II interferons. These results provide a general insight into the interaction between CHIKV and its mammalian host

Astrodynamical Space Test of Relativity using Optical Devices I (ASTROD I) - A class-M fundamental physics mission proposal for Cosmic Vision 2015-2025: 2010 Update

This paper on ASTROD I is based on our 2010 proposal submitted for the ESA call for class-M mission proposals, and is a sequel and an update to our previous paper [Experimental Astronomy 23 (2009) 491-527; designated as Paper I] which was based on our last proposal submitted for the 2007 ESA call. In this paper, we present our orbit selection with one Venus swing-by together with orbit simulation. In Paper I, our orbit choice is with two Venus swing-bys. The present choice takes shorter time (about 250 days) to reach the opposite side of the Sun. We also present a preliminary design of the optical bench, and elaborate on the solar physics goals with the radiation monitor payload. We discuss telescope size, trade-offs of drag-free sensitivities, thermal issues and present an outlook. ASTROD I is a planned interplanetary space mission with multiple goals. The primary aims are: to test General Relativity with an improvement in sensitivity of over 3 orders of magnitude, improving our understanding of gravity and aiding the development of a new quantum gravity theory; to measure key solar system parameters with increased accuracy, advancing solar physics and our knowledge of the solar system; and to measure the time rate of change of the gravitational constant with an order of magnitude improvement and the anomalous Pioneer acceleration, thereby probing dark matter and dark energy gravitationally. It is envisaged as the first in a series of ASTROD missions. ASTROD I will consist of one spacecraft carrying a telescope, four lasers, two event timers and a clock. Two-way, two-wavelength laser pulse ranging will be used between the spacecraft in a solar orbit and deep space laser stations on Earth, to achieve the ASTROD I goals.Comment: 15 pages, 11 figures, 1 table, based on our 2010 proposal submitted for the ESA call for class-M mission proposals, a sequel and an update to previous paper [Experimental Astronomy 23 (2009) 491-527] which was based on our last proposal submitted for the 2007 ESA call, submitted to Experimental Astronom

Human Immunodeficiency Virus Impairs Reverse Cholesterol Transport from Macrophages

Several steps of HIV-1 replication critically depend on cholesterol. HIV infection is associated with profound changes in lipid and lipoprotein metabolism and an increased risk of coronary artery disease. Whereas numerous studies have investigated the role of anti-HIV drugs in lipodystrophy and dyslipidemia, the effects of HIV infection on cellular cholesterol metabolism remain uncharacterized. Here, we demonstrate that HIV-1 impairs ATP-binding cassette transporter A1 (ABCA1)-dependent cholesterol efflux from human macrophages, a condition previously shown to be highly atherogenic. In HIV-1–infected cells, this effect was mediated by Nef. Transfection of murine macrophages with Nef impaired cholesterol efflux from these cells. At least two mechanisms were found to be responsible for this phenomenon: first, HIV infection and transfection with Nef induced post-transcriptional down-regulation of ABCA1; and second, Nef caused redistribution of ABCA1 to the plasma membrane and inhibited internalization of apolipoprotein A-I. Binding of Nef to ABCA1 was required for down-regulation and redistribution of ABCA1. HIV-infected and Nef-transfected macrophages accumulated substantial amounts of lipids, thus resembling foam cells. The contribution of HIV-infected macrophages to the pathogenesis of atherosclerosis was supported by the presence of HIV-positive foam cells in atherosclerotic plaques of HIV-infected patients. Stimulation of cholesterol efflux from macrophages significantly reduced infectivity of the virions produced by these cells, and this effect correlated with a decreased amount of virion-associated cholesterol, suggesting that impairment of cholesterol efflux is essential to ensure proper cholesterol content in nascent HIV particles. These results reveal a previously unrecognized dysregulation of intracellular lipid metabolism in HIV-infected macrophages and identify Nef and ABCA1 as the key players responsible for this effect. Our findings have implications for pathogenesis of both HIV disease and atherosclerosis, because they reveal the role of cholesterol efflux impairment in HIV infectivity and suggest a possible mechanism by which HIV infection of macrophages may contribute to increased risk of atherosclerosis in HIV-infected patients

MICROSCOPE mission: first results of a space test of the equivalence principle

According to the weak equivalence principle, all bodies should fall at the same rate in a gravitational field. The MICROSCOPE satellite, launched in April 2016, aims to test its validity at the 10−15 precision level, by measuring the force required to maintain two test masses (of titanium and platinum alloys) exactly in the same orbit. A nonvanishing result would correspond to a violation of the equivalence principle, or to the discovery of a new long-range force. Analysis of the first data gives δ(Ti,Pt)=[−1±9(stat)±9(syst)]×10−15 (1σ statistical uncertainty) for the titanium-platinum Eötvös parameter characterizing the relative difference in their free-fall accelerations