Determination of thermodynamic parameters of Xerocomus chrysenteron lectin interactions with N-acetylgalactosamine and Thomsen-Friedenreich antigen by isothermal titration calorimetry

BACKGROUND: Lectins are carbohydrate-binding proteins which potentially bind to cell surface glycoconjugates. They are found in various organisms including fungi. A lectin from the mushroom Xerocomus chrysenteron (XCL) has been isolated recently. It shows insecticidal activity and has antiproliferative properties. RESULTS: As the monosaccharide binding specificity is an important determinant of lectin function, we determined the affinity of XCL for the galactose moiety. Isothermal titration calorimetry studies revealed a dissociation constant K(d )of 5.2 μM for the XCL:N-acetylgalactosamine interaction at 27degreesC. Higher affinities were observed at lower temperatures and higher osmotic pressures. The dissociation constant was five hundred times higher for the disaccharide beta-D-Gal(1–3)-D-GalNAc, Thomsen-Friedenreich (TF) antigen (Kd of 0.94 μM). By using fetuin and asialofetuin in interaction with the XCL, we revealed its ability to recognize the Thomsen-Friedenreich motif on glycoproteins. CONCLUSION: The XCL antiproliferative effect and the TF antigen specificity presented in this work suggest that XCL and ABL may have similar binding mechanisms. The recent structure determination of these two proteins lead us to analyse these interactions in the light of our thermodynamic data. The understanding of this type of interaction may be a useful tool for the regulation of cell proliferation

Protein encapsulation in liposomes: efficiency depends on interactions between protein and phospholipid bilayer.

BACKGROUND: We investigated the encapsulation mechanism of enzymes into liposomes. The existing protocols to achieve high encapsulation efficiencies are basically optimized for chemically stable molecules. Enzymes, however, are fragile and encapsulation requires in addition the preservation of their functionality. Using acetylcholinesterase as a model, we found that most protocols lead to a rapid denaturation of the enzyme with loss in the functionality and therefore inappropriate for such an application. The most appropriate method is based on lipid film hydration but had a very low efficiency. RESULTS: To improve it and to propose a standard procedure for enzyme encapsulation, we separate each step and we studied the effect of each parameter on encapsulation: lipid and buffer composition and effect of the different physical treatment as freeze-thaw cycle or liposomes extrusion. We found that by increasing the lipid concentration, increasing the number of freeze-thaw cycles and enhancing the interactions of the enzyme with the liposome lipid surface more than 40% of the initial total activity can be encapsulated. CONCLUSION: We propose here an optimized procedure to encapsulate fragile enzymes into liposomes. Optimal encapsulation is achieved by induction of a specific interaction between the enzyme and the lipid surface

Méthode de Monte Carlo et synthèse d'images : application à des milieux diffusants en transfert radiatif

International audienceL'objectif principal de cette étude est la prise en compte de la diffusion multiple pour le calcul de transferts radiatifs dans des géométries complexes. Pour traiter la diffusion, un algorithme basé sur la méthode de Monte Carlo a été développé et implémenté dans un environnement issu d'un code de synthèse d'images existant. A l'aide de propriétés d'invariance de la statistique des chemins de diffusion multiple, nous avons validé cette approche dans le cas de trois géométries tridimensionnelles usuelles : sphère, cylindre et pyramide

Liposome retention in size exclusion chromatography

BACKGROUND: Size exclusion chromatography is the method of choice for separating free from liposome-encapsulated molecules. However, if the column is not presaturated with lipids this type of chromatography causes a significant loss of lipid material. To date, the mechanism of lipid retention is poorly understood. It has been speculated that lipid binds to the column material or the entire liposome is entrapped inside the void. RESULTS: Here we show that intact liposomes and their contents are retained in the exclusion gel. Retention depends on the pore size, the smaller the pores, the higher the retention. Retained liposomes are not tightly fixed to the beads and are slowly released from the gels upon direct or inverted eluent flow, long washing steps or column repacking. Further addition of free liposomes leads to the elution of part of the gel-trapped liposomes, showing that the retention is transitory. Trapping reversibility should be related to a mechanism of partitioning of the liposomes between the stationary phase, water-swelled polymeric gel, and the mobile aqueous phase. CONCLUSION: Retention of liposomes by size exclusion gels is a dynamic and reversible process, which should be accounted for to control lipid loss and sample contamination during chromatography

Current-Based Detection of Mechanical Unbalance in an Induction Machine Using Spectral Kurtosis with Reference

This article explores the design, on-line, of an electrical machine’s healthy reference by means of statistical tools. The definition of a healthy reference enables the computation of normalized fault indicators whose value is independent of the system’s characteristics. This is a great advantage when diagnosing a broad range of systems with different power, coupling, inertia, load, etc. In this paper, an original method called spectral kurtosis with reference is presented in order to designa system’s healthy reference. Its principle is first explained on asynthetic signal. This approach is then evaluated for mechanicalunbalance detection in an induction machine using the stator currents instantaneous frequency. The normalized behaviour ofthe proposed indicator is then confirmed for different operatingconditions and its robustness with respect to load variationsis demonstrated. Finally, the advantages of using a statisticalindicator based on a healthy reference compared to a raw faultsignature are discussed

Statistic-based method to monitor belt transmission looseness through motor phase currents

Belt–pulley systems are widely used in the industry due to their high efficiency and their low cost. However, only few works exist about the monitoring of their degradation. This paper details the impact of belt looseness on electrical measurements under steady and transient state in order to identify spectral signatures. This analysis enlightens the advantage of the transient state to detect belt looseness because it exacerbates belt slip. An innovative methodology is then proposed based on the application of a square-wave speed reference in order to monitor belt looseness. A statistical-based indicator is defined from the phase currents in order to automatically detect drifting of the indicator. A normalization process is also applied to increase the detection robustness. The proposed indicatoris evaluated on a 30-kW induction machine and a direct current machine coupled with two trapezoidal belts for three speed and four load conditions. It reaches very good results with almost 90% correct detections for 1% false alarms. These results are way better than those obtained with a classic spectral analysis during the steady state. Moreover, results demonstrate that higher load conditions are more accurate for the monitoring of belt looseness. Index Terms—Belt–pulley systems, fault diagnosis, induction motors (IMs), robustness, spectral analysis, statistical-based indicator, torque–speed segmentation

Hippocampal neuroligin-2 overexpression leads to reduced aggression and inhibited novelty reactivity in rats

Disturbances of the excitation/inhibition (E/I) balance in the brain were recently suggested as potential factors underlying disorders like autism and schizophrenia resulting in associated behavioral alterations including changes in social and emotional behavior as well as abnormal aggression. Neuronal cell adhesion molecules (nCAMs) and mutations in these genes were found to be strongly implicated in the pathophysiology of these disorders. Neuroligin2 (nlgn2) is a postsynaptic cell adhesion molecule, which is predominantly expressed at inhibitory synapses and required for synapse specification and stabilization. Changes in the expression of nlgn2 were shown to result in alterations of social behavior as well as altered inhibitory synaptic transmission, hence modifying the E/I balance. In our study, we focused on the role of nlgn2 in the dorsal hippocampus in the regulation of emotional and social behaviors. To this purpose, we injected an AAV construct overexpressing nlgn2 in the hippocampus of rats and investigated the effects on behavior and on markers for the E/I ratio. We could show an increase in GAD65, a GABA-synthesizing protein in neuronal terminals, and furthermore, reduced exploration of novel stimuli and less offensive behavior. Our data suggest nlgn2 in the hippocampus to be strongly implicated in maintaining the E/I balance in the brain and thereby modulating social and emotional behavior

The interplay of conditional NCAM-knockout and chronic unpredictable stress leads to increased aggression in mice

Abstract The neural cell adhesion molecule (NCAM) is a key regulator of brain plasticity. Substantial evidence indicates that NCAM is down-regulated by exposure to sustained stress and chronic stress seems to lead to increased aggression. In addition, constitutional NCAM deletion in mice has been shown to lead to increased intermale aggression and altered emotionality Forebrain-specific postnatal NCAM knockout was previously shown to impair cognitive function, particularly when animals were exposed to subchronic stress, but the effects on emotional and social behavior remain unclear. In this study, we investigated the potential interplay of a forebrain-specific postnatal NCAM deletion and exposure to different lengths of repeated stress (i.e., subchronic: 14 days; chronic: 29 days) on aggressive and emotional behavior. Our results show that postnatal deletion of NCAM in the forebrain leads to increased aggression and altered emotionality depending on the duration of stress, whereas conditional NCAM knockout has no basal impact on these behaviors. These findings support the involvement of NCAM in the regulation of emotional and aggressive behaviors, suggesting that diminished NCAM expression might be a critical vulnerability factor for the development of these behavioral alterations under repeated exposure to stres

Genome analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea

Sclerotinia sclerotiorum and Botrytis cinerea are closely related necrotrophic plant pathogenic fungi notable for their wide host ranges and environmental persistence. These attributes have made these species models for understanding the complexity of necrotrophic, broad host-range pathogenicity. Despite their similarities, the two species differ in mating behaviour and the ability to produce asexual spores. We have sequenced the genomes of one strain of S. sclerotiorum and two strains of B. cinerea. The comparative analysis of these genomes relative to one another and to other sequenced fungal genomes is provided here. Their 38–39 Mb genomes include 11,860–14,270 predicted genes, which share 83% amino acid identity on average between the two species. We have mapped the S. sclerotiorum assembly to 16 chromosomes and found large-scale co-linearity with the B. cinerea genomes. Seven percent of the S. sclerotiorum genome comprises transposable elements compared t

Validating canopy clumping retrieval methods using hemispherical photography in a simulated Eucalypt forest

The so-called clumping factor (Ω) quantifies deviation from a random 3D distribution of material in a vegetation canopy and therefore characterises the spatial distribution of gaps within a canopy. Ω is essential to convert effective Plant or Leaf Area Index into actual LAI or PAI, which has previously been shown to have a significant impact on biophysical parameter retrieval using optical remote sensing techniques in forests, woodlands, and savannas. Here, a simulation framework was applied to assess the performance of existing in situ clumping retrieval methods in a 3D virtual forest canopy, which has a high degree of architectural realism. The virtual canopy was reconstructed using empirical data from a Box Ironbark Eucalypt forest in Eastern Australia. Hemispherical photography (HP) was assessed due to its ubiquity for indirect LAI and structure retrieval. Angular clumping retrieval method performance was evaluated using a range of structural configurations based on varying stem distribution and LAI. The CLX clumping retrieval method (Leblanc et al., 2005) with a segment size of 15° was the best performing clumping method, matching the reference values to within 0.05 Ω on average near zenith. Clumping error increased linearly with zenith angle to > 0.3 Ω (equivalent to a 30% PAI error) at 75° for all structural configurations. At larger zenith angles, PAI errors were found to be around 25–30% on average when derived from the 55–60° zenith angle. Therefore, careful consideration of zenith angle range utilised from HP is recommended. We suggest that plot or site clumping factors should be accompanied by the zenith angle used to derive them from gap size and gap size distribution methods. Furthermore, larger errors and biases were found for HPs captured within 1 m of unrepresentative large tree stems, so these situations should be avoided in practice if possible