Cooperative Retraction of Bundled Type IV Pili Enables Nanonewton Force Generation

The causative agent of gonorrhea, Neisseria gonorrhoeae, bears retractable filamentous appendages called type IV pili (Tfp). Tfp are used by many pathogenic and nonpathogenic bacteria to carry out a number of vital functions, including DNA uptake, twitching motility (crawling over surfaces), and attachment to host cells. In N. gonorrhoeae, Tfp binding to epithelial cells and the mechanical forces associated with this binding stimulate signaling cascades and gene expression that enhance infection. Retraction of a single Tfp filament generates forces of 50–100 piconewtons, but nothing is known, thus far, on the retraction force ability of multiple Tfp filaments, even though each bacterium expresses multiple Tfp and multiple bacteria interact during infection. We designed a micropillar assay system to measure Tfp retraction forces. This system consists of an array of force sensors made of elastic pillars that allow quantification of retraction forces from adherent N. gonorrhoeae bacteria. Electron microscopy and fluorescence microscopy were used in combination with this novel assay to assess the structures of Tfp. We show that Tfp can form bundles, which contain up to 8–10 Tfp filaments, that act as coordinated retractable units with forces up to 10 times greater than single filament retraction forces. Furthermore, single filament retraction forces are transient, whereas bundled filaments produce retraction forces that can be sustained. Alterations of noncovalent protein–protein interactions between Tfp can inhibit both bundle formation and high-amplitude retraction forces. Retraction forces build over time through the recruitment and bundling of multiple Tfp that pull cooperatively to generate forces in the nanonewton range. We propose that Tfp retraction can be synchronized through bundling, that Tfp bundle retraction can generate forces in the nanonewton range in vivo, and that such high forces could affect infection

Sl-ARF4,an Auxin Response Factor involved in the control of sugar metabolism during tomato fruit development

Successful completion of fruit developmental programs depends on the interplay between multiple phytohormones. However,besides ethylene, the impact of other hormones on fruit quality traits remains elusive. A previous study has shown that downregulation of SlARF4, a member of the tomato (Solanum lycopersicum) auxin response factor (ARF) gene family, results in a darkgreen fruit phenotype with increased chloroplasts (Jones et al., 2002). This study further examines the role of this auxin transcriptional regulator during tomato fruit development at the level of transcripts, enzyme activities, and metabolites. It is noteworthy that the dark-green phenotype of antisense SlARF4-suppressed lines is restricted to fruit, suggesting that SlARF4 controls chlorophyll accumulation specifically in this organ. The SlARF4 underexpressing lines accumulate more starch at early stages of fruit development and display enhanced chlorophyll content and photochemical efficiency, which is consistent with the idea that fruit photosynthetic activity accounts for the elevated starch levels. SlARF4 expression is high in pericarp tissues of immature fruit and then undergoes a dramatic decline at the onset of ripening concomitant with the increase in sugar content. The higher starch content in developing fruits of SlARF4 down-regulated lines correlates with the up-regulation of genes and enzyme activities involved in starch biosynthesis, suggesting their negative regulation by SlARF4. Altogether, the data uncover the involvement of ARFs in the control of sugar content, an essential feature of fruit quality, and provide insight into the link between auxin signaling, chloroplastic activity, and sugar metabolism in developing fruit

Beyond Post-release Mortality: Inferences on Recovery Periods and Natural Mortality From Electronic Tagging Data for Discarded Lamnid Sharks

Accurately characterizing the biology of a pelagic shark species is critical when assessing its status and resilience to fishing pressure. Natural mortality (M) is well known to be a key parameter determining productivity and resilience, but also one for which estimates are most uncertain. While M can be inferred from life history, validated direct estimates are extremely rare for sharks. Porbeagle (Lamna nasus) and shortfin mako (Isurus oxyrinchus) are presently overfished in the North Atlantic, but there are no directed fisheries and successful live release of bycatch is believed to have increased. Understanding M, post-release mortality (PRM), and variables that affect mortality are necessary for management and effective bycatch mitigation. From 177 deployments of archival satellite tags, we inferred mortality events, characterized physiological recovery periods following release, and applied survival mixture models to assess M and PRM. We also evaluated covariate effects on the duration of any recovery period and PRM to inform mitigation. Although large sample sizes involving extended monitoring periods (>90 days) would be optimal to directly estimate M from survival data, it was possible to constrain estimates and infer probable values for both species. Furthermore, the consistency of M estimates with values derived from longevity information suggests that age determination is relatively accurate for these species. Regarding bycatch mitigation, our analyses suggest that juvenile porbeagle are more susceptible to harm during capture and handling, that keeping lamnid sharks in the water during release is optimal, and that circle hooks are associated with longer recovery periods for shortfin mako.En prens

Overconfidence in Labor Markets

This chapter reviews how worker overconfidence affects labor markets. Evidence from psychology and economics shows that in many situations, most people tend to overestimate their absolute skills, overplace themselves relative to others, and overestimate the precision of their knowledge. The chapter starts by reviewing evidence for overconfidence and for how overconfidence affects economic choices. Next, it reviews economic explanations for overconfidence. After that, it discusses research on the impact of worker overconfidence on labor markets where wages are determined by bargaining between workers and firms. Here, three key questions are addressed. First, how does worker overconfidence affect effort provision for a fixed compensation scheme? Second, how should firms design compensation schemes when workers are overconfident? In particular, will a compensation scheme offered to an overconfident worker have higher-or lower-powered incentives than that offered to a worker with accurate self-perception? Third, can worker overconfidence lead to a Pareto improvement? The chapter continues by reviewing research on the impact of worker overconfidence on labor markets where workers can move between firms and where neither firms nor workers have discretion over wage setting. The chapter concludes with a summary of its main findings and a discussion of avenues for future research

Étude structurale et mécanismes de formation de complexes amylose-ligand (Influence sur la rétention de composés d'arôme dans une matrice amylacée)

L'objet de ce travail est d'étudier les mécanismes de complexation de l'amylose par des composés d'arômes et la nature des structures formées, afin d'appréhender l'influence de l'amidon sur la rétention d'arômes dans une matrice alimentaire (génoise). La structure semi-cristalline et la morphologie des 3 types de complexe obtenus ont été approchées par diffraction des rayons X, MET et modélisation moléculaire. La localisation des composés d'arômes et leur teneur dans les zones amorphes et cristallines ont été déterminées par utilisation couplée de spectroscopie FT-IR et d'hydrolyses acides. L'étude des cinétiques de cristallisation/fusion par AED et FT-IR a permis de proposer 2 mécanismes de complexation et de déterminer la stabilité thermique des différents complexes étudiés. L'ensemble des résultats suggèrent une réelle implication des complexes d'amylose sur la rétention des arômes dans une matrice alimentaire. Ceci a pu être vérifié au cours de la préparation d'une génoise.This work aims at studying the complexing of amylose by a series of aroma compounds in order to grasp the influence of starch on the trapping of flavours in food matrices as sponge-cakes. The semi-crystalline structure (crystallinity, polymorphic type, inclusion of the aroma compound in and between amylose helices) of the resulting 3 types of complex and the related morphology were assessed by X-Ray scattering, TEM. and molecular modelling. The location and the respective amount of aroma compounds in amorphous and crystalline regions stoichiometry were determined by using both FT-IR spectroscopy and acid hydrolysis. The kinetics of crystallisation and the thermal stability were investigated by DSC and FT-IR . It allowed to propose 2 different complexing mechanisms. The results suggest a real involvement of amylose complexes on the retention of flavours in real food matrices. This point was verified during sponge-cake baking.NANTES-BU Sciences (441092104) / SudocSudocFranceF

Metabolic acclimation to hypoxia revealed by metabolite gradients in melon fruit

International audienceA metabolomics approach using 1H NMR and GC–MS profiling of primary metabolites and quantification of adenine nucleotides with luciferin bioluminescence was employed to investigate the spatial changes of metabolism in melon fruit. Direct 1H NMR profiling of juice collected from different locations in the fruit flesh revealed several gradients of metabolites, e.g. sucrose, alanine, valine, GABA or ethanol, with increase in concentrations from the periphery to the center of the fruit. GC–MS profiling of ground samples revealed gradients for metabolites not detected using 1H NMR, including pyruvic and fumaric acids. The quantification of adenine nucleotides highlighted a strong decrease in both ATP and ADP ratios and the adenylate energy charge from the periphery to the center of the fruit. These concentration patterns are consistent with an increase in ethanol fermentation due to oxygen limitation and were confirmed by observed changes in alanine and GABA concentrations, as well as other markers of hypoxia in plants. Ethanol content in melon fruit can affect organoleptic properties and consumer acceptance. Understanding how and when fermentation occurred can help to manage the culture and limit ethanol production

Molecules

ε-Viniferin is a resveratrol dimer that possesses antioxidant or anti-inflammatory activities. However little is known about the metabolism of this oligostilbene. This study was thus undertaken as a first approach to identify and characterize the metabolites of ε-viniferin and to describe the kinetic profile of their appearance in humans and rats. The glucuronides and sulfates of ε-viniferin were first obtained by chemical hemi-synthesis and were fully characterized by UPLC-MS and NMR spectroscopy. Then, ε-viniferin was incubated with human or rat S9 liver fractions that led to the formation of four glucuronoconjugates and four sulfoconjugates. In both species, ε-viniferin was subjected to an intense metabolism as 70 to 80% of the molecule was converted to glucuronides and sulfates. In humans, the hepatic clearance of ε-viniferin (V/K) for glucuronidation and sulfation were 4.98 and 6.35 µL/min/mg protein, respectively, whereas, in rats, the hepatic clearance for glucuronidation was 20.08 vs. 2.59 µL/min/mg protein for sulfation. In humans, three major metabolites were observed: two glucuronides and one sulfate. By contrast, only one major glucuronide was observed in rats. This strong hepatic clearance of ε-viniferin in human and rat could explain its poor bioavailability and could help to characterize its active metabolites

Modelling central metabolic fluxes by constraint-based optimization reveals metabolic reprogramming of developing Solanum lycopersicum (tomato) fruit.

Modelling of metabolic networks is a powerful tool to analyse the behaviour of developing plant organs, including fruits. Guided by our current understanding of heterotrophic metabolism of plant cells, a medium-scale stoichiometric model, including the balance of co-factors and energy, was constructed in order to describe metabolic shifts that occur through the nine sequential stages of Solanum lycopersicum (tomato) fruit development. The measured concentrations of the main biomass components and the accumulated metabolites in the pericarp, determined at each stage, were fitted in order to calculate, by derivation, the corresponding external fluxes. They were used as constraints to solve the model by minimizing the internal fluxes. The distribution of the calculated fluxes of central metabolism were then analysed and compared with known metabolic behaviours. For instance, the partition of the main metabolic pathways (glycolysis, pentose phosphate pathway, etc.) was relevant throughout fruit development. We also predicted a valid import of carbon and nitrogen by the fruit, as well as a consistent CO2 release. Interestingly, the energetic balance indicates that excess ATP is dissipated just before the onset of ripening, supporting the concept of the climacteric crisis. Finally, the apparent contradiction between calculated fluxes with low values compared with measured enzyme capacities suggest a complex reprogramming of the metabolic machinery during fruit development. With a powerful set of experimental data and an accurate definition of the metabolic system, this work provides important insight into the metabolic and physiological requirements of the developing tomato fruits