Dickeya dadantii pectic enzymes necessary for virulence are also responsible for activation of the Arabidopsis thaliana innate immune system.

Soft-rot diseases of plants attributed to Dickeya dadantii result from lysis of the plant cell wall caused by pectic enzymes released by the bacterial cell by a type II secretion system (T2SS). Arabidopsis thaliana can express several lines of defence against this bacterium. We employed bacterial mutants with defective envelope structures or secreted proteins to examine early plant defence reactions. We focused on the production of AtrbohD-dependent reactive oxygen species (ROS), callose deposition and cell death as indicators of these reactions. We observed a significant reduction in ROS and callose formation with a bacterial mutant in which genes encoding five pectate lyases (Pels) were disrupted. Treatment of plant leaves with bacterial culture filtrates containing Pels resulted in ROS and callose production, and both reactions were dependent on a functional AtrbohD gene. ROS and callose were produced in response to treatment with a cellular fraction of a T2SS-negative mutant grown in a Pels-inducing medium. Finally, ROS and callose were produced in leaves treated with purified Pels that had also been shown to induce the expression of jasmonic acid-dependent defence genes. Pel catalytic activity is required for the induction of ROS accumulation. In contrast, cell death observed in leaves infected with the wild-type strain appeared to be independent of a functional AtrbohD gene. It was also independent of the bacterial production of pectic enzymes and the type III secretion system (T3SS). In conclusion, the work presented here shows that D. dadantii is recognized by the A. thaliana innate immune system through the action of pectic enzymes secreted by bacteria at the site of infection. This recognition leads to AtrbohD-dependent ROS and callose accumulation, but not cell death.

Complex nonequilibrium dynamics of stacked polystyrene films deep in the glassy state.

We investigate the kinetics of enthalpy recovery in stacked glassy polystyrene (PS) films with thickness from 30 to 95 nm over a wide temperature range below the glass transition temperature (Tg). We show that the time evolution toward equilibrium exhibits two mechanisms of recovery, in ways analogous to bulk PS. The fast mechanism, allowing partial enthalpy recovery toward equilibrium, displays Arrhenius temperature dependence with low activation energy, whereas the slow mechanism follows pronounced super-Arrhenius temperature dependence. In comparison to bulk PS, the time scales of the two mechanisms of recovery are considerably shorter and decreasing with the film thickness. Scaling of the equilibration times at various thicknesses indicates that the fast mechanism of recovery is compatible with the free volume holes diffusion model. Conversely, the slow mechanism of recovery appears to be accelerated with decreasing thickness more than predicted by the model and, therefore, its description requires additional ingredients. The implications, from both a fundamental and technological viewpoint, of the ability of thin polymer films to densify in relatively short time scales are discussed.

Reaching the ideal glass transition by aging polymer films.

Searching for the ideal glass transition, we exploit the ability of glassy polymer films to explore low energy states in remarkably short time scales. We use 30 nm thick polystyrene (PS) films, which in the supercooled state basically display the bulk polymer equilibrium thermodynamics and dynamics. We show that in the glassy state, this system exhibits two mechanisms of equilibrium recovery. The faster one, active well below the kinetic glass transition temperature (Tg), allows massive enthalpy recovery. This implies that the 'fictive' temperature (Tf) reaches values as low as the predicted Kauzmann temperature (TK) for PS. Once the thermodynamic state corresponding to Tf = TK is reached, no further decrease of enthalpy is observed. This is interpreted as a signature of the ideal glass transition.

Direct evidence of two equilibration mechanisms in glassy polymers.

We investigated the kinetics of enthalpy recovery of several glass-forming polymers at temperatures significantly below the glass transition temperature (Tg) and for aging times up to one year. We find a double-step recovery at relatively low aging temperatures for the longest investigated aging times. The enthalpy recovered after the two-step decay approximately equals that expected by extrapolation from the melt. The two-step enthalpy recovery indicates the presence of two time scales for glass equilibration. The equilibration time of the first recovery step exhibits relatively weak temperature dependence, whereas that of the second step possesses pronounced temperature dependence, compatible with the Vogel-Fulcher-Tammann behavior. These results, while leaving open the question of the divergence of the relaxation time and that of a thermodynamic singularity at a finite temperature, reveal a complex scenario of glassy dynamics.

Time dependence of the segmental relaxation time of poly(vinyl acetate)-silica nanocomposites.

The aging-time dependence of the segmental relaxation time of poly(vinyl acetate) (PVAc) in the glassy state is investigated in the bulk polymer and its nanocomposites with silica (SiO(2)). These systems present identical segmental dynamics, when this is probed in the equilibrium supercooled liquid by broadband dielectric spectroscopy. An acceleration of the physical aging process of PVAc with SiO(2) was detected by monitoring the enthalpy recovery through differential scanning calorimetry. The segmental relaxation time during physical aging, followed by means of BDS, has been shown to increase more rapidly the higher the SiO(2) concentration in PVAc is. Thermally stimulated depolarization current experiments show that this is the case over the whole probed glassy state. This means that nanocomposites displaying a relatively slow segmental mobility evolve toward equilibrium more rapidly than the bulk. Furthermore, despite the faster increase in the relaxation time with aging time, so-called self-retardation, the nanocomposites and their bulk counterpart reach the same values of equilibrium relaxation time. These findings not only confirm the assumption of identical equilibrium dynamics even in the aging regime for all nanocomposites and bulk polymers, proposed in previous works, but also highlight the fact that the physical aging rate is not determined solely by the polymer segmental dynamics, the amount of interface being an additional relevant parameter.

Cortical and subcortical projections to primary visual cortex in anophthalmic, enucleated and sighted mice.

The purpose of this study was to identify and compare the afferent projections to the primary visual cortex in intact and enucleated C57BL/6 mice and in ZRDCT/An anophthalmic mice. Early loss of sensory-driven activity in blind subjects can lead to activations of the primary visual cortex by haptic or auditory stimuli. This intermodal activation following the onset of blindness is believed to arise through either unmasking of already present cortical connections, sprouting of novel cortical connections or enhancement of intermodal cortical connections. Studies in humans have similarly demonstrated heteromodal activation of visual cortex following relatively short periods of blindfolding. This suggests that the primary visual cortex in normal sighted subjects receives afferents, either from multisensory association cortices or from primary sensory cortices dedicated to other modalities. Here cortical afferents to the primary visual cortex were investigated to determine whether the visual cortex receives sensory input from other modalities, and whether differences exist in the quantity and/or the structure of projections found in sighted, enucleated and anophthalmic mice. This study demonstrates extensive direct connections between the primary visual cortex and auditory and somatosensory areas, as well as with motor and association cortices in all three animal groups. This suggests that information from different sensory modalities can be integrated at early cortical stages and that visual cortex activations following visual deprivations can partly be explained by already present intermodal corticocortical connections.

Easy-dispersible poly(glycidyl phenyl ether)-functionalized graphene sheets obtained by reaction of "living" anionic polymer chains.

Excellent dispersion of functionalized graphene (FG) sheets in polystyrene was achieved relying on the reaction of "living" poly(glycidyl phenyl ether) chains onto graphene sheets. The physical aging of polystyrene was substantially accelerated by the presence of FG sheets at low filler content, retaining film transparency and increasing the electrical conductivity.

Free volume holes diffusion to describe physical aging in poly(mehtyl methacrylate)/silica nanocomposites.

The spontaneous thermodynamically driven densification, the so-called physical aging, of glassy poly(mehtyl methacrylate) (PMMA) and its nanocomposites with silica has been described by means of the free volume holes diffusion model. This mechanism is able to account for the partial decoupling between physical aging and segmental dynamics of PMMA in nancomposites. The former has been found to be accelerated in PMMA/silica nanocomposites in comparison to "bulk" PMMA, whereas no difference between the segmental dynamics of bulk PMMA and that of the same polymer in nanocomposites has been observed. Thus, the rate of physical aging also depends on the amount of interface polymer/nanoparticles, where free volume holes disappear after diffusing through the polymer matrix. The free volume holes diffusion model is able to nicely capture the phenomenology of the physical aging process with a structure dependent diffusion coefficient.

MtPM25 is an atypical hydrophobic late embryogenesis-abundant protein that dissociates cold and desiccation-aggregated proteins.

Late embryogenesis-abundant (LEA) proteins are one of the components involved in desiccation tolerance (DT) by maintaining cellular structures in the dry state. Among them, MtPM25, a member of the group 5 is specifically associated with DT in Medicago truncatula seeds. Its function is unknown and its classification as a LEA protein remains elusive. Here, evidence is provided that MtPM25 is a hydrophobic, intrinsically disordered protein that shares the characteristics of canonical LEA proteins. Screening protective activities by testing various substrates against freezing, heating and drying indicates that MtPM25 is unable to protect membranes but able to prevent aggregation of proteins during stress. Prevention of aggregation was also found for the water soluble proteome of desiccation-sensitive radicles. This inhibition was significantly higher than that of MtEM6, one of the most hydrophilic LEA protein associated with DT. Moreover, when added after the stress treatment, MtPM25 is able to rapidly dissolve aggregates in a non-specific manner. Sorption isotherms show that when it is unstructured, MtPM25 absorbs up to threefold more water than MtEM6. MtPM25 is likely to act as a protective molecule during drying and plays an additional role as a repair mechanism compared with other LEA proteins.

Arabidopsis thaliana expresses multiple lines of defense to counterattack Erwinia chrysanthemi.

Many taxonomically diverse plant species are attacked by Erwinia chrysanthemi, a member of the causal agents of soft-rotting diseases. Symptom development is due to the collective action of pectin-degrading enzymes secreted by the bacterium through a type II secretion system (T2SS). Using Arabidopsis thaliana as a susceptible host, we show that plants respond to E. chrysanthemi 3937 by expressing cell-wall reactions, production of an oxidative burst, and activation of salicylic acid (SA) and jasmonic acid (JA) or ethylene (ET) signaling pathways. We found that the oxidative burst is mainly generated via the expression of the AtrbohD gene, constitutes a barrier of resistance to bacterial attack, and acts independently of the SA-mediated response. To determine the importance of T2SS-secreted proteins in elicitation of these defenses, we used a T2SS deficient mutant and purified enzymatic preparations of representative members of strain 3937 pectate lyase activity. The T2SS-secreted proteins were responsible only partially for the activation of SA and JA or ET signaling pathways observed after infection with the wild-type bacterium and were not involved in the expression of other identified defense reactions. Our study shows the differential role played by pectate lyases isoenzymes in this process and highlights the complexity of the host immune network, which is finely controlled by the bacterium.

Effects of 4-hydroxytamoxifen, raloxifene and ICI 182 780 on survival of uterine cancer cell lines in the presence and absence of exogenous estrogens.

The impact of 17beta-estradiol and antiestrogens on uterine cancer cells is poorly understood. The aim of this study was to determine the impact of 17beta-estradiol, 4-hydroxytamoxifen, raloxifene and ICI 182 780 on the cell proliferation of six uterine cancer cell lines: HeLa, HEC-1-A, KLE, RL-95-2, Ishikawa and EN-1078D. The effects of these compounds on the cell proliferation of the six uterine cancer cell lines were studied in the presence and absence of estrogens (phenol red and serum deprivation of sex steroids). In a general manner, 17beta-estradiol and 4-hydroxytamoxifen showed similarities in their effects whereas raloxifene showed a different pattern of cell proliferation (agonistic and antagonistic) and ICI 182 780 had antagonistic activity. In the presence and absence of estrogens, we observed that each cell line had diverse expression of ERalpha, ERbeta, GPR30 and REA. GPR30 mRNA expression was significantly reduced in a serum/phenol-free medium. REA mRNA expression was not influenced by the media. Results demonstrated the importance of removing phenol red and the use of deprived serum when studying uterine cancer cells in relationship with 17beta-estradiol and antiestrogens. The affinity of each compound to the binding of ERalpha and ERbeta was very similar with the exception of raloxifene that had a preference for ERalpha binding. Akt phosphorylation/activity was reduced in cells cultured in a phenol red- and steroid-free culture medium indicating that the presence of steroids in the culture media can influence the activity of this survival pathway. Our results suggest that the expression of ERalpha, ERbeta and GPR30 are influenced by sex steroids and might play a role in the response of cells to 17beta-estradiol and antiestrogens but are not the only factors involved in this process.