Susceptibility and possible resistance mechanisms in the palm species Phoenix dactylifera, Chamaerops humilis and Washingtonia filifera against Rhynchophorus ferrugineus (Olivier, 1790) (Coleoptera: Curculionidae).

Rhynchophorus ferrugineus, known as the Red Palm Weevil (RPW), is reported as a pest of up to 40 palm species. However, the susceptibility degree and the defense mechanisms of these species against this weevil are still poorly known. In Europe, the RPW is a major pest of Phoenix canariensis while other palm species, including the congeneric Phoenix dactylifera, seem to be less suitable hosts for this insect. The aim of our study was to compare the defensive response of P. dactylifera, Chamaerops humilis and Washingtonia filifera against R. ferrugineus and try to define the mechanisms of resistance that characterize these species. Bioassays were carried out to evaluate the mortality induced on RPW larvae by extracts from the leaf rachis of the studied palm species. Tests at semi-field scale were also conducted, based either on forced palm infestation, with larvae of RPW, or on natural infestation, with adult females. Rachis extracts from C. humilis and W. filifera caused 100% larval mortality after 2 days of exposure, while extracts of P. dactylifera did not impair larval survival. Independently of the effect of the leaf extracts, the weevils were unable to naturally infest the three palm species, although larval survival was high after forced infestation of the plants. We concluded that the observed lack of infestation of P. dactylifera by RPW is due to factors other than antibiosis. In W. filifera and C. humilis, although the presence of antixenosis mechanisms cannot be excluded, resistance to R. ferrugineus seems to rely on the presence of antibiosis compounds.

Thermomechanical transitions of egg-ceramide monolayers.

Ceramides have unique biophysical properties. Their high melting temperature and their ability to form lateral domains have converted ceramides into the paradigm of rigid lipids. Here, using shear surface rheology of egg-ceramide Langmuir monolayers, a solid to fluid transition was evidenced as a vanishing shear rigidity at lower temperatures than the lipid melting temperature. Such a mechanical transition, which depends on the lipid lateral pressure, was found in a broad range temperature (40-50 °C). The solid to fluid transition was correlated to a LC to LC+LE phase transition, as confirmed by BAM experiments. Interestingly, together with the softening transition, a supercooling process compatible with a glassy behavior was found upon freezing. A new phase scenario is then depicted that broadens the mechanical behavior of natural ceramides. The phase diversity of ceramides might have important implications in their physiological roles.

Membrane reconstitution of FtsZ-ZipA complex inside giant spherical vesicles made of E. coli lipids: large membrane dilation and analysis of membrane plasticity.

During the division process of Escherichia coli, the globular protein FtsZ is early recruited at the constriction site. The Z-ring, based on FtsZ filaments associated to the inner cell membrane, has been postulated to exert constriction forces. Membrane anchoring is mediated by ZipA, an essential transmembrane protein able to specifically bind FtsZ. In this work, an artificial complex of FtsZ-ZipA has been reconstituted at the inner side of spherical giant unilamellar vesicles made of E. coli lipids. Under these conditions, FtsZ polymerization, triggered when a caged GTP analogue is UV-irradiated, was followed by up to 40% vesicle inflation. The homogeneous membrane dilation was accompanied by the visualization of discrete FtsZ assemblies at the membrane. Complementary rheological data revealed enhanced elasticity under lateral dilation. This explains why vesicles can undergo large dilations in the regime of mechanical stability. A mechanical role for FtsZ polymers as promoters of membrane softening and plasticization is hypothesized.

Variance sum rule for entropy production.

Entropy production is the hallmark of nonequilibrium physics, quantifying irreversibility, dissipation, and the efficiency of energy transduction processes. Despite many efforts, its measurement at the nanoscale remains challenging. We introduce a variance sum rule (VSR) for displacement and force variances that permits us to measure the entropy production rate σ in nonequilibrium steady states. We first illustrate it for directly measurable forces, such as an active Brownian particle in an optical trap. We then apply the VSR to flickering experiments in human red blood cells. We find that σ is spatially heterogeneous with a finite correlation length, and its average value agrees with calorimetry measurements. The VSR paves the way to derive σ using force spectroscopy and time-resolved imaging in living and active matter.

Langmuir polymer films: recent results and new perspectives.

Langmuir polymer films (LPFs) are very interesting systems to probe quasi-two dimensional dynamics. Although adsorbed on water, the substrate is fluid enough to avoid irreversible pinning at adsorption sites, as with solid substrates. LPFs in dense states can exhibit a high degree of metastability, however reproducible measurements can be performed on films which have not been previously compressed. The shear rheology is one of the most active fields of research, especially because it allows investigation of flow behaviour in LPFs, thus of possible reptation motion in semidilute films under good solvent conditions. It also allows probing glassy behaviour in dense films under poor solvent conditions. In this perspective article, we review the recent literature and discuss unpublished results on the dynamics of the glass transition, recently observed in these quasi-2D systems at low temperatures. We conclude by listing new problems and open questions emerging from this research area.

Regulation of chemokine responses in intestinal epithelial cells by stress and Toxoplasma gondii infection.

Infection with Toxoplasma gondii induces chemokine up-regulation in several cell types. Here, we investigated the role of stress products (norepinephrine, NE) on chemokine production in mouse intestinal epithelial cells (IECs). Purified IECs were used to determine the expression levels of chemokines by real-time PCR. There was significantly increased expression in CCL2, CCL3, CCL5, CXCL2, CXCL9 and CXCL10 in IECs following peroral infection with T. gondii (INF) on day eight post-infection (PI) compared to infected mice subjected to cold-water stress (INF+CWS). In vitro studies using the MODE-K cell line showed increased chemokine mRNA and protein expression in infected but not in cells exposed to parasite antigen. Down-regulation of chemokine expression was more pronounced when active infection was used in combination with NE. Chemokine receptor expression was increased in IECs isolated from INF and decreased in the INF+CWS group. In MODE-K cells, there was decreased mRNA expression of chemokine receptors when incubated with ß-adrenergic antagonists. Neither, adrenergic antagonists blocked the effect of infection on chemokine receptor expression. Cold-water stress was able to decrease expression of chemokines and their receptors in IECs in vivo and in vitro. Cold-water stress-mediated modulation of innate intestinal responses are beneficial in C57BL/6 mice during T. gondii infection.

Amorphous freezing in two dimensions: from soft coils to rigid particles.

The topic of the gel transition in two dimensions is revisited by considering data on the shear elasticity of Langmuir monolayers of different spherical objects. Amorphous freezing can be associated to structural percolation in a lattice able to resist shear stresses. The shear modulus and its dependence on the packing fraction are found to strongly depend on the details of the interaction potential and largely differ from expectations for entropic networks. This behaviour can be interpreted in terms of more elaborated percolation theories including central forces and bond-bending forces.

Dissipative curvature fluctuations in bilayer vesicles: Coexistence of pure-bending and hybrid curvature-compression modes.

We have studied the relaxation dynamics of shape fluctuations in unilamellar lipid vesicles by neutron spin echo (NSE). The presence of a hybrid curvature-compression mode coexisting with the usual bending one has been revealed in the experimental relaxation functions at high q . Differently to the conventional relaxation approximately q (3) typical for bending modes, the hybrid mode was found to relax as approximately q (2) , which is compatible with a dissipation mechanism arising from intermonolayer friction. Complementary data obtained from flickering spectroscopy (FS) in giant unilamellar vesicles confirm the existence of both modes coexisting together. By combining NSE and FS data we have depicted the experimental bimodal dispersion diagram, which is found compatible with theoretical predictions for reliable values of the material parameters. From the present data two conventional dynamical methods (NSE and FS) have been shown to be suitable for measuring intermonolayer friction coefficients in bilayer vesicles.

Membrane stress and torque induced by Frank's nematic textures: A geometric perspective using surface-based constraints.

An elastic membrane with embedded nematic molecules is considered as a model of anisotropic fluid membrane with internal ordering. By considering the geometric coupling between director field and membrane curvature, the nematic texture is shown to induce anisotropic stresses additional to Canham-Helfrich elasticity. Building upon differential geometry, analytical expressions are found for the membrane stress and torque induced by splaying, twisting, and bending of the nematic director as described by the Frank energy of liquid crystals. The forces induced by prototypical nematic textures are visualized on the sphere and on cylindrical surfaces.

Regulation of toll-like receptors in intestinal epithelial cells by stress and Toxoplasma gondii infection.

Intestinal epithelial cells (IECs) form a barrier between invading microorganisms and the underlying host tissues. IECs express toll-like receptors (TLRs) that recognize specific molecular signatures on microbes, which activate intracellular signalling pathways leading to production of proinflammatory cytokines and chemokines. Stress hormones play an important role in modulation of proinflammatory cytokines and down-regulation of immune responses. Here we demonstrated that expression levels of TLR-2, TLR-4, TLR-9 and TLR-11 were significantly increased in mouse IECs following infection with Toxoplasma gondii on day 8 postinfection. In contrast, expression of TLRs was significantly decreased in infected mice subjected to cold water stress (CWS + INF). Expression of TLR-9 and TLR-11 in the mouse MODE-K cell line was significantly increased after infection. Expression of TLR-9 and TLR-11 in cells exposed to norepinephrine (NE) and parasites was significantly decreased when compared to cells exposed to parasites only. A significant increase was observed in SIGIRR, a negative regulator of TLRs in the CWS + INF group when compared to the INF group. Stress components were able to decrease expression levels of TLRs in IECs, decrease parasite load, and increase expression of a negative regulator thereby ameliorating intestinal inflammatory responses commonly observed during per oral T. gondii infection in C57BL/6 mice.

Bovine anaplasmosis and tick-borne pathogens in cattle of the Galapagos Islands.

A cross-sectional study was conducted to determine the species of Anaplasma spp. and estimate its prevalence in cattle of the three main cattle-producing Galapagos Islands (Santa Cruz, San Cristóbal and Isabela) using indirect PCR assays, genetic sequencing and ELISA. Ticks were also collected from cattle and scanned for 47 tick-borne pathogens in a 48 × 48 real-time PCR chip. A mixed effects logistic regression was performed to identify potential risk factors explaining Anaplasma infection in cattle. A. phagocytophilum was not detected in any of the tested animals. Genetic sequencing allowed detection of A. platys-like strains in 11 (36.7%) of the 30 Anaplasma spp.-positive samples analysed. A. marginale was widespread in the three islands with a global between-herd prevalence of 100% [89; 100]95% CI and a median within-herd prevalence of 93%. A significant association was found between A. marginale infection and age with higher odds of being positive for adults (OR = 3.3 [1.2; 9.9]95% Bootstrap CI ). All collected ticks were identified as Rhipicephalus microplus. A. marginale, Babesia bigemina, Borrelia theileri and Francisella-like endosymbiont were detected in tick pools. These results show that the Galapagos Islands are endemic for A. marginale.

Permeability modes in fluctuating lipid membranes with DNA-translocating pores.

Membrane pores can significantly alter not only the permeation dynamics of biological membranes but also their elasticity. Large membrane pores able to transport macromolecular contents represent an interesting model to test theoretical predictions that assign active-like (non-equilibrium) behavior to the permeability contributions to the enhanced membrane fluctuations existing in permeable membranes [Maneville et al. Phys. Rev. Lett. 82, 4356 (1999)]. Such high-amplitude active contributions arise from the forced transport of solvent and solutes through the open pores, which becomes even dominant at large permeability. In this paper, we present a detailed experimental analysis of the active shape fluctuations that appear in highly permeable lipid vesicles with large macromolecular pores inserted in the lipid membrane, which are a consequence of transport permeability events occurred in an osmotic gradient. The experimental results are found in quantitative agreement with theory, showing a remarkable dependence with the density of membrane pores and giving account of mechanical compliances and permeability rates that are compatible with the large size of the membrane pore considered. The presence of individual permeation events has been detected in the fluctuation time-series, from which a stochastic distribution of the permeation events compatible with a shot-noise has been deduced. The non-equilibrium character of the membrane fluctuations in a permeation field, even if the membrane pores are mere passive transporters, is clearly demonstrated. Finally, a bio-nano-technology outlook of the proposed synthetic concept is given on the context of prospective uses as active membrane DNA-pores exploitable in gen-delivery applications based on lipid vesicles.

Carbohydrate-binding plasma proteins from the gastropod Biomphalaria glabrata: strain specificity and the effects of trematode infection.

As lectins are believed to mediate non-self-recognition in molluscs, carbohydrate-binding proteins (CBP) from the circulating plasma of the gastropod Biomphalaria glabrata were harvested by affinity chromatography using six different monosaccharides as ligands. Pools of plasma were derived from B. glabrata of either the M line strain, which is susceptible to infection with the PR1 strain of the digenetic trematode Schistosoma mansoni, or from the 13-16-R1 strain, which is resistant to infection. For each strain, plasma was obtained from control snails and from snails exposed to infection 1 or 8 days previously with S. mansoni or the related digenean, Echinostoma paraensei, which is able to develop in either host strain. For control snails, only minor interstrain differences were noted. In M line snails exposed 8 days previously to either parasite, marked changes in CBP populations were observed. Only E. paraensei infections produced comparable alterations in 13-16-R1 snails. The most conspicuous changes noted were the increased production of 80-120 kDa CBP in both strains, 150-210 kDa in 13-16-R1 snails, and 190-210 kDa in M line snails. The results demonstrate 1) interstrain differences in CBP, particularly following exposure to trematodes; 2) that infection provokes increased production and diversity of CBP that bind with greater affinity to the columns; and 3) that snails of the same strain respond differently to the two parasites used.

The developmental expression of cysteine proteinases in Schistosoma mansoni.

In this study several probes were used to identify and characterize cysteine proteinases (CP) from Schistosoma mansoni developmental stages. Proteinase activity was detected in all developmental stages using fluorogenic substrates. Specific activity was 4- to 11-fold higher against CBZ-phe-arg-AFC than against CBZ-arg-arg-AFC. A 32 kDa S. mansoni CP (Sm32) was identified in all stages by the radiolabelled active site CP inhibitor, CBZ-[125I]tyr-ala-CHN2. A second CP, of 31 kDa (Sm31) was only detected in adult worms, primarily female worms and late schistosomula. Monoclonal antibodies for Sm32 and Sm31 reacted with homologous CP only in adult worms and late schistosomula. This study defines the developmental expression of CP activity in S. mansoni.

Low molecular weight immunosuppressors secreted by adult Nematospiroides dubius.

Adult Nematospiroides dubius excretory-secretory products (ES) were collected from worms cultured in vitro, separated by sodium dodecyl sulphate--polyacrylamide gel electrophoresis (SDS-PAGE) into four fractions (FI-IV), electroeluted and assessed for their ability to inhibit the proliferation of mouse lymphocytes stimulated by mitogens in vitro. The proliferation of mitogen- and ES-stimulated mouse spleen lymphocytes from normal and infected mice was inhibited by low mol. wt ES F-IV (less than 26,000).

Proteolytic enzymes in excretory-secretory products from adult Nematospiroides dubius.

Excretory-secretory products (ES), collected from in vitro cultures of adult Nematospiroides dubius, were examined for proteolytic enzyme activity. ES enzymes had a pH optimum of 8.0 and their activity was sensitive to serine-proteinase inhibitors. Three SDS-resistant proteases were identified in ES at molecular weight (mol. wt) 200,000, 105,000 and 48,000 by incorporating substrates into the matrices of sodium dodecyl sulfate-polyacrylamide electrophoresis (SDS-PAGE) gels.

Immunity in mice vaccinated with a molecular weight 60,000 glycoprotein secreted by adult Nematospiroides dubius.

A mol. wt 60,000 glycoprotein was purified from adult Nematospiroides dubius excretory-secretory products (ES) by polyacrylamide gel electrophoresis and electroelution and used to vaccinate BCF1 mice. This molecule is also released from the surface of the parasite and can be cleaved by pepsin. Mice immunized twice with ES 60,000 in Alum adjuvant harboured similar numbers of N. dubius as did control mice but voided 40% fewer parasite eggs in their faeces. The mol. wt 60,000 component in ES and from the surface of the parasite appears to influence how the parasite reproduces and may be important to its survival.

Survival of Trypanosoma musculi in the kidneys of chronically infected mice: kidney form reproduction and immunological reactions.

Trypanosoma musculi are detected in the blood of the mouse host within 3-5 days after infection. Peak parasitemia is reached within 10 days and parasites persist at a plateau level for 2-3 wk. There are no intracellular stages and the flagellates are eliminated from the peripheral blood within 4 wk. However, trypanosomes persist in the vasa recta of the kidneys and may be present for the life of the host. Infection provides lifelong resistance to reinfection. Kidney forms (KF) of T. musculi were isolated and studied to define their morphology, reproductive activity, and serological reactivity. Dividing epimastigotes and trypomastigote stages were present in kidneys. Multinucleate and rosette forms were common. Measurements of the coefficient of variation of the KF confirmed that the trypanosomes were actively reproducing. Direct immunofluorescence reactions with rabbit antimouse IgG + IgA + IgM detected antibodies on bloodstream forms (BSF) but not on the KF. However, indirect immunofluorescence tests using antisera collected from mice during early and late phases of the infections showed that KF were capable of reacting with antibodies. The reproductive activity displayed by the KF appears to be responsible for the continuous replacement of the trypanosomes that are killed by the immune responses of the resistant mice when they leave the vasa recta. Solute concentrations of the blood within the vasa recta appear to prevent antibodies from complexing with surface antigens of the parasites. These capillaries provide nutrients that allow the trypanosomes to reproduce and persist unaffected by the host immune responses.

Production of heterogeneous carbohydrate-binding proteins by the host snail Biomphalaria glabrata following exposure to Echinostoma paraensei and Schistosoma mansoni.

Hemolymph lectins may play an important role in the internal defense responses of gastropods to parasites. Two groups of known carbohydrate-binding polypeptides, of 150-220 kDa (designated as group 1 molecules, or G1M) and of 75-130 kDa (group 2 molecules, or G2M), were harvested from pooled plasma samples of Biomphalaria glabrata using affinity chromatography and examined using 2-dimensional electrophoresis. Plasma samples were derived from control snails or snails exposed 8 days earlier to the trematodes Echinostoma paraensei or Schistosoma mansoni. Plasma of control and S. mansoni-exposed snails contained little or no G1M, whereas plasma from E. paraensei-infected snails contained G1M covering a broad pI spectrum. G2M resolved as 1-2 isoforms in control plasma and up to 4 relatively faint isoforms in plasma from S. mansoni-exposed snails, and as 5-6 resolvable isoforms in plasma from E. paraensei-infected snails. Plasma from individual snails contained as many as 5 G2M polypeptides following exposure to E. paraensei. Exposure to trematode larvae stimulated production by B. glabrata of increased abundance and diversity of carbohydrate-binding proteins. The 2 trematode species provoked different responses, and 2 B. glabrata strains studied (M line and 13-16-R1 strains) differed from one another in their responses.

Trypanosoma musculi survival in the kidneys of chronically infected mice: kidney form ultrastructure, surface characteristics, and serological interactions.

After elimination of Trypanosoma musculi from the general circulation by the immune responses of infected mice, the animals are resistant to reinfection. Yet, parasites survive in the vasa recta of the kidneys for the life of these mice. These kidney forms (KF) actively reproduce in an environment that provides the necessary nutrients and appears to prevent their elimination from these capillaries by the hosts' immune responses. Comparative studies conducted with KF and the bloodstream forms (BSF) indicate that, although both forms appear to be similar morphologically at the ultrastructural level, they differ in their surface reactivities with lectins and tolerance to various pH and solute concentrations. Although antibodies are not detected on the surfaces of KF, urea levels approximating those in the vasa recta dissociate antibody from the surfaces of BSF. The data suggest that parasites found in the vasa recta of these chronically infected mice differ from the BSF and are protected from the humoral and cell-mediated immune responses of the murine hosts by the concentrated solutes present in these capillaries. The KF may be killed by these same immune effector mechanisms upon leaving the capillaries of the kidneys and, therefore, not be found in the general circulation of these chronically infected immune hosts.