This work describes a coordinate and comprehensive view on the time course of the alterations occurring at the level of the cell wall during adaptation of a yeast cell population to sudden exposure to a sub-lethal stress induced by acetic acid. Acetic acid is a major inhibitory compound in industrial bioprocesses and a widely used preservative in foods and beverages. Results indicate that yeast cell wall resistance to lyticase activity increases during acetic acid-induced growth latency, corresponding to yeast population adaptation to sudden exposure to this stress. This response correlates with: (i) increased cell stiffness, assessed by atomic force microscopy (AFM); (ii) increased content of cell wall β-glucans, assessed by fluorescence microscopy, and (iii) slight increase of the transcription level of the GAS1 gene encoding a β-1,3-glucanosyltransferase that leads to elongation of (1→3)-β-d-glucan chains. Collectively, results reinforce the notion that the adaptive yeast response to acetic acid stress involves a coordinate alteration of the cell wall at the biophysical and molecular levels. These alterations guarantee a robust adaptive response essential to limit the futile cycle associated to the re-entry of the toxic acid form after the active expulsion of acetate from the cell interior.

An important loss mechanism in organic electroluminescent devices is exciton quenching by polarons. Gradual electrochemical doping of various conjugated polymer films enabled the determination of the doping density dependence of photoluminescence quenching. Electrochemical doping was achieved by contacting the film with a solid electrochemical gate and an injecting contact. A sharp reduction in photoluminescence was observed for doping densities between 10 18 and 10 19 cm −3 . The doping density dependence is quantitatively modeled by exciton diffusion in a homogeneous density of polarons followed by either Förster resonance energy transfer or charge transfer. Both mechanisms need to be considered to describe polaron-induced exciton quenching. Thus, to reduce exciton-polaron quenching in organic optoelectronic devices, both mechanisms must be prevented by reducing the exciton diffusion, the spectral overlap, the doping density, or a combination thereof.

Summary Diacylglycerol kinases (DGKs) play a major role in the production of phosphatidic acid (PtdOH) and were implicated in endomembrane trafficking and signalling cascades. In plants, the role of DGKs is less clear, as PtdOH seems to arise mostly from phospholipase D activity. Here, we investigated the function of the Arabidopsis gene encoding DGK4, which is highly expressed in pollen. In vitro, pollen tubes from homozygous dgk4 plants showed normal morphology, but reduced growth rate and altered stiffness and adhesion properties (revealed by atomic force microscopy). In vivo, dgk4 pollen was able to fertilize wild‐type ovules, but self‐pollination in dgk4 plants led to fewer seeds and shorter siliques. Phenotypic analysis revealed that the dgk4 mutation affects not only the male germ line but also the vegetative tissue. DGK4‐green fluorescent protein fusion imaging revealed a cytosolic localization with a slightly higher signal in the subapical or apical region. dgk4 pollen tubes were found to exhibit perturbations in membrane recycling, and lipid analysis revealed a minor increase of PtdOH concomitant with decreased phosphatidylcholine, compared with wild‐type. In vitro, DGK4 was found to exhibit kinase and guanylyl cyclase activity. Quantitative PCR data revealed downregulation of genes related to actin dynamics and phosphoinositide metabolism in mutant pollen, but upregulation of the DGK6 isoform. Altogether, these results are discussed considering a role of DGK4 in signalling cross‐talk.

Acetic acid is a major inhibitory compound in several industrial bioprocesses, in particular in lignocellulosic yeast biorefineries. Cell envelope remodeling, involving cell wall and plasma membrane composition, structure and function, is among the mechanisms behind yeast adaptation and tolerance to stress. Pdr18 is a plasma membrane ABC transporter of the pleiotropic drug resistance family and a reported determinant of acetic acid tolerance mediating ergosterol transport. This study provides evidence for the impact of Pdr18 expression in yeast cell wall during adaptation to acetic acid stress. The time-course of acetic-acid-induced transcriptional activation of cell wall biosynthetic genes (FKS1, BGL2, CHS3, GAS1) and of increased cell wall stiffness and cell wall polysaccharide content in cells with the PDR18 deleted, compared to parental cells, is reported. Despite the robust and more intense adaptive response of the pdr18Δ population, the stress-induced increase of cell wall resistance to lyticase activity was below parental strain levels, and the duration of the period required for intracellular pH recovery from acidification and growth resumption was higher in the less tolerant pdr18Δ population. The ergosterol content, critical for plasma membrane stabilization, suffered a drastic reduction in the first hour of cultivation under acetic acid stress, especially in pdr18Δ cells. Results revealed a crosstalk between plasma membrane ergosterol content and cell wall biophysical properties, suggesting a coordinated response to counteract the deleterious effects of acetic acid.

Water menisci wet all sorts of cavities, produce among the most intense forces at the nanoscale and play a role in many physical and chemical processes. The physical properties of these menisci are therefore relevant to understand a multitude of phenomena at the nanoscale where these are involved. Here, using a force feedback microscope, we directly measured the capillary condensation time of a water meniscus, by approaching two surfaces at different speeds and monitoring the relative position of the surfaces at the instant the meniscus is formed.

The performance of a custom atomic force microscope for grazing-incidence X-ray experiments on hydrated soft and biological samples is presented.

Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR). A single recessive mutation, the deletion of phenylalanine 508 (F508del), causes severe CF and resides on 70% of mutant chromosomes. Disorganization of the actin cytoskeleton has been previously reported in relation to the CF phenotype. In this work, we aimed to understand this alteration by means of Atomic Force Microscopy and Force Feedback Microscopy investigation of mechanical properties of cystic fibrosis bronchial epithelial (CFBE) cells stably transduced with either wild type (wt-) or F508del-CFTR. We show here that the expression of mutant CFTR causes a decrease in the cell's apparent Young modulus as compared to the expression of the wt protein.

We report the observation of a transition in the dynamical properties of water nano-menicus which dramatically change when probed at different time scales. Using a AFM mode that we name Force Feedback Microscopy, we observe this change in the simultaneous measurements, at different frequencies, of the stiffness G'(N/m), the dissipative coefficient G"(kg/sec) together with the static force. At low frequency we observe a negative stiffness as expected for capillary forces. As the measuring time approaches the microsecond, the dynamic response exhibits a transition toward a very large positive stiffness. When evaporation and condensation gradually lose efficiency, the contact line progressively becomes immobile. This transition is essentially controlled by variations of Laplace pressure.Visco-elastic properties of water nanobridges[1] at very different time scales, have never been investigated despite ubiquitous presence of capillarity. Associated forces are among the most intense at nanoscales with important consequences in soils and granular media. Interest in dynamical properties is immediately raised if one considers interacting surfaces with roughness scales down to nanometer. Even at moderate speeds, such as v=1m/s, characteristic times of surface interaction down to microsecond appear in these conditions. Our measurements approaching these time scales, further strengthen the relevance of the dynamical properties to describe how real surfaces interact and are certainly of crucial importance in numerous AFM experiments [2]. We here report measurements of dynamical properties of a water nanobridge for a continuous range of the surface gap and a frequency bandwidth up to 0.1 MHz. We identify two regimes: one is the thermodynamical equilibrium; the second is out of equilibrium. Evaporation and condensation of water molecules between the liquid and the gas phase ensures that the nano-meniscus curvature is the one at thermodynamical equilibrium(2H=-1/r k ) where 2H is the water bridge curvature and r k is the Kelvin radius. At time short enough, molecule exchanges between the liquid and the gas phase are no longer efficient and the water nanobridge is led to acquire a constant volume. The liquid bridge relaxation time is the time needed for the bridge to adapt its shape as required by thermodynamical equilibrium, when its length h is abruptly changed by δh. This is controlled by molecular transport through diffusion mechanisms in gas phase. This relaxation time τ can be estimated as in SFA context, see Ref.[3]. τ = 2γρr 2 ln(R/ρ)/P sat r k 2 D