Determining physical properties of the cell cortex

Actin and myosin assemble into a thin layer of a highly dynamic network underneath the membrane of eukaryotic cells. This network generates the forces that drive cell and tissue-scale morphogenetic processes. The effective material properties of this active network determine large-scale deformations and other morphogenetic events. For example,the characteristic time of stress relaxation (the Maxwell time)in the actomyosin sets the time scale of large-scale deformation of the cortex. Similarly, the characteristic length of stress propagation (the hydrodynamic length) sets the length scale of slow deformations, and a large hydrodynamic length is a prerequisite for long-ranged cortical flows. Here we introduce a method to determine physical parameters of the actomyosin cortical layer (in vivo). For this we investigate the relaxation dynamics of the cortex in response to laser ablation in the one-cell-stage {\it C. elegans} embryo and in the gastrulating zebrafish embryo. These responses can be interpreted using a coarse grained physical description of the cortex in terms of a two dimensional thin film of an active viscoelastic gel. To determine the Maxwell time, the hydrodynamic length and the ratio of active stress and per-area friction, we evaluated the response to laser ablation in two different ways: by quantifying flow and density fields as a function of space and time, and by determining the time evolution of the shape of the ablated region. Importantly, both methods provide best fit physical parameters that are in close agreement with each other and that are similar to previous estimates in the two systems. We provide an accurate and robust means for measuring physical parameters of the actomyosin cortical layer.It can be useful for investigations of actomyosin mechanics at the cellular-scale, but also for providing insights in the active mechanics processes that govern tissue-scale morphogenesis.Comment: 17 pages, 4 figure

Pulsatory Patterns in Active Fluids

We show that pulsatory patterns arise in thin active films in which two chemical species regulate active stress. The regulating species diffuse within the film and are advected by self-generated flows resulting from active stress gradients. Spontaneous pulsatory patterns emerge when the following conditions are met: (i) the fast-diffusing species up-regulates and the slow-diffusing species down-regulates active stress, or (ii) the active stress up-regulator turns over faster compared to the active stress down-regulator. Our study, motivated by pulsatory patterns in the actomyosin cortex in cells and tissues, provides a simple generic mechanism for oscillatory patterns in active fluids

Polarization of PAR Proteins by Advective Triggering of a Pattern-Forming System

In the Caenorhabditis elegans zygote, a conserved network of partitioning-defective (PAR) polarity proteins segregates into an anterior and a posterior domain, facilitated by flows of the cortical actomyosin meshwork. The physical mechanisms by which stable asymmetric PAR distributions arise from transient cortical flows remain unclear. We present evidence that PAR polarity arises from coupling of advective transport by the flowing cell cortex to a multistable PAR reaction-diffusion system. By inducing transient PAR segregation, advection serves as a mechanical trigger for the formation of a PAR pattern within an otherwise stably unpolarized system. We suggest that passive advective transport in an active and flowing material may be a general mechanism for mechanochemical pattern formation in developmental systems

HIV Infection of Naturally Occurring and Genetically Reprogrammed Human Regulatory T-cells

A T-cell subset, defined as CD4(+)CD25(hi) (regulatory T-cells [Treg cells]), was recently shown to suppress T-cell activation. We demonstrate that human Treg cells isolated from healthy donors express the HIV-coreceptor CCR5 and are highly susceptible to HIV infection and replication. Because Treg cells are present in very few numbers and are difficult to expand in vitro, we genetically modified conventional human T-cells to generate Treg cells in vitro by ectopic expression of FoxP3, a transcription factor associated with reprogramming T-cells into a Treg subset. Overexpression of FoxP3 in naïve human CD4(+) T-cells recapitulated the hyporesponsiveness and suppressive function of naturally occurring Treg cells. However, FoxP3 was less efficient in reprogramming memory T-cell subset into regulatory cells. In addition, FoxP3-transduced T-cells also became more susceptible to HIV infection. Remarkably, a portion of HIV-positive individuals with a low percentage of CD4(+) and higher levels of activated T-cells have greatly reduced levels of FoxP3(+)CD4(+)CD25(hi) T-cells, suggesting disruption of the Treg cells during HIV infection. Targeting and disruption of the T-cell regulatory system by HIV may contribute to hyperactivation of conventional T-cells, a characteristic of HIV disease progression. Moreover, the ability to reprogram human T-cells into Treg cells in vitro will greatly aid in decoding their mechanism of suppression, their enhanced susceptibility to HIV infection, and the unique markers expressed by this subset

Excitonic photoluminescence in symmetric coupled double quantum wells subject to an external electric field

The effect of an external electric field F on the excitonic photoluminescence (PL) spectra of a symmetric coupled double quantum well (DQW) is investigated both theoretically and experimentally. We show that the variational method in a two-particle electron-hole wave function approximation gives a good agreement with measurements of PL on a narrow DQW in a wide interval of F including flat-band regime. The experimental data are presented for an MBE-grown DQW consisting of two 5 nm wide GaAs wells, separated by a 4 monolayers (MLs) wide pure AlAs central barrier, and sandwiched between Ga_{0.7}Al_{0.3}As layers. The bias voltage is applied along the growth direction. Spatially direct and indirect excitonic transitions are identified, and the radius of the exciton and squeezing of the exciton in the growth direction are evaluated variationally. The excitonic binding energies, recombination energies, oscillator strengths, and relative intensities of the transitions as functions of the applied field are calculated. Our analysis demonstrates that this simple model is applicable in case of narrow DQWs not just for a qualitative description of the PL peak positions but also for the estimation of their individual shapes and intensities.Comment: 5 pages, 4 figures (accepted in Phys. Rev. B

Engagement of Fusiform Cortex and Disengagement of Lateral Occipital Cortex in the Acquisition of Radiological Expertise

The human visual pathways that are specialized for object recognition stretch from lateral occipital cortex (LO) to the ventral surface of the temporal lobe, including the fusiform gyrus. Plasticity in these pathways supports the acquisition of visual expertise, but precisely how training affects the different regions remains unclear. We used functional magnetic resonance imaging to measure neural activity in both LO and the fusiform gyrus in radiologists as they detected abnormalities in chest radiographs. Activity in the right fusiform face area (FFA) correlated with visual expertise, measured as behavioral performance during scanning. In contrast, activity in left LO correlated negatively with expertise, and the amount of LO that responded to radiographs was smaller in experts than in novices. Activity in the FFA and LO correlated negatively in experts, whereas in novices, the 2 regions showed no stable relationship. Together, these results suggest that the FFA becomes more engaged and left LO less engaged in interpreting radiographic images over the course of training. Achieving expert visual performance may involve suppressing existing neural representations while simultaneously developing others

Mechanochemical feedback regulates the dynamics of the PAR system in C. elegans zygotes

The interplay between regulatory biochemistry and cell mechanics is critical for a broad range of morphogenetic changes. Cell mechanics can induce transport via growth and flow-fields, which in turn affect concentration-fields of regulators. Such systems exhibit an intrinsic feedback-architecture between regulators of cell mechanics and mechanical deformation. While we anticipate that this feedback between biochemistry and cell mechanics is widespread in Morphogenesis, there are few examples that are studied with respect to their potential for generating spatiotemporal patterns. Here we establish at a quantitative level that PAR polarization of C. elegans zygotes represents a coupled mechanochemical system. Using Fluorescence Recovery After Photobleaching (FRAP) and RNA interference (RNAi), we first demonstrate that the biochemistry in form of the PAR domains feeds back on the mechanics by establishing and maintaining a non-muscle myosin II (nmy-2) gradient. Additionally, we characterize the effect of the polarity cue associated with the centrosome of the male pronucleus on the local myosin concentration at the posterior pole. We show that it induces a reduction in myosin concentration and thereby triggers the onset of cortical flows. Furthermore we measure the spatiotemporal profile of the anterior and posterior PAR concentration, the myosin II concentration and the induced flow-field. Finally, we capture the feedback-architecture of the coupled actomyosin – PAR system in a quantitative model, based on coupling a thin film active fluid description of cortical mechanics [1] to a reaction-diffusion PAR patterning system [2]. We show that this mathematical model can quantitatively recapitulate the spatiotemporal profile of PAR polarity establishment. Furthermore, we demonstrate that the model predicts the existence of a threshold in cortical flow velocity, which separates the nonpolarizing and the polarizing regime and confirm the existence of this threshold velocity in the living C. elegans zygote

Two-Dimensional Molecular Patterning by Surface-Enhanced Zn-Porphyrin Coordination

In this contribution, we show how zinc-5,10,15,20-meso-tetradodecylporphyrins (Zn-TDPs) self-assemble into stable organized arrays on the surface of graphite, thus positioning their metal center at regular distances from each other, creating a molecular pattern, while retaining the possibility to coordinate additional ligands. We also demonstrate that Zn-TDPs coordinated to 3-nitropyridine display a higher tendency to be adsorbed at the surface of highly oriented pyrolytic graphite (HOPG) than noncoordinated ones. In order to investigate the two-dimensional (2D) self-assembly of coordinated Zn-TDPs, solutions with different relative concentrations of 3-nitropyridine and Zn-TDP were prepared and deposited on the surface of HOPG. STM measurements at the liquid-solid interface reveal that the ratio of coordinated Zn-TDPs over noncoordinated Zn-TDPs is higher at the n-tetradecane/HOPG interface than in n-tetradecane solution. This enhanced binding of the axial ligand at the liquid/solid interface is likely related to the fact that physisorbed Zn-TDPs are better binding sites for nitropyridines.

Improving outcome over time of percutaneous coronary interventions in unstable angina

AbstractOBJECTIVEThis study was performed to evaluate the recent changes in the outcome of coronary interventions in patients with unstable angina (UA).BACKGROUNDAn early invasive strategy has not been shown to be superior to conservative treatment in patients with UA. Earlier studies had utilized older technology. Interventional approaches have changed in the recent past, but to our knowledge, no large studies have addressed the impact of these changes on the outcome of coronary interventions.METHODSWe analyzed the in-hospital and intermediate-term outcome in 7,632 patients with UA who underwent coronary interventions in the last two decades. The study population was divided into three groups: group 1, n = 2,209 who had coronary intervention from 1979 to 1989; group 2, n = 2,212 with interventions from 1990 to 1993; and group 3, n = 3,211 treated from 1994 to 1998.RESULTSGroup 2 and 3 patients were older and sicker compared with group 1 patients. The clinical success improved significantly in group 3 (94.1%) compared with group 2 (87%) and group 1 (76.5%) (p < 0.001). There was a significant reduction in in-hospital mortality, Q-wave myocardial infarction and need for emergency bypass surgery in group 3 compared with the earlier groups. One-year event-free survival was also significantly higher in the recent group compared with the earlier groups: 77% in group 3, 70% in group 2 and 74% in group 1 (p < 0.001). With the use of multivariate models to adjust for clinical and angiographic variables, treatment during the most recent era was found to be independently associated with improved in-hospital and intermediate-term outcomes.CONCLUSIONSThere has been significant improvement in the in-hospital and intermediate-term outcome of coronary interventions in patients with UA in recent years; newer trials comparing conservative and invasive strategies are therefore needed