Live 3D imaging and mapping of shear stresses within tissues using incompressible elastic beads

To investigate the role of mechanical constraints in morphogenesis and development, we develop a pipeline of techniques based on incompressible elastic sensors. These techniques combine the advantages of incompressible liquid droplets, which have been used as precise in situ shear stress sensors, and of elastic compressible beads, which are easier to tune and to use. Droplets of a polydimethylsiloxane (PDMS) mix, made fluorescent through specific covalent binding to a rhodamin dye, are produced by a microfluidics device. The elastomer rigidity after polymerization is adjusted to the tissue rigidity. Its mechanical properties are carefully calibrated in situ, for a sensor embedded in a cell aggregate submitted to uniaxial compression. Thelocal shear stress tensor is retrieved from the sensor shape, accurately reconstructed through an active contour method. In vitro, within cell aggregates, and in vivo, in the prechordal plate of the Zebrafish embryo during gastrulation,our pipeline of techniques demonstrates its efficiency to directly measure the three dimensional shear stress repartition within a tissue.

Release of HIV-1 particles from the viral compartment in macrophages requires an associated cytoskeleton and is driven by mechanical constraints

A defining feature of HIV-1 replication in macrophages is that viral assembly occurs at the limiting membrane of a compartment often named VCC (virus-containing compartments) that is connected to the extracellular medium. The newly formed viral progeny pinches of the membrane and accumulates in the lumen of the VCC. While HIV budding has been extensively studied, very little is known about how viral particles present in the lumen of VCC are released in the extracellular medium. Here we show that the actin dynamics are critical for this process by combining ultrastructural analyses, time-lapse microscopy and perturbations of the actin cytoskeleton. We found that jasplakinolide, which stabilizes actin fibres, inhibited viral release from HIV-1-infected macrophages, but not from infected HeLa cells. Furthermore, in jasplakinolide-treated macrophages, VCC became scattered and no longer co-localized with the integrin CD18, nor the phosphorylated form of the focal adhesion kinase PYK2. Inhibition of PYK2 activity in infected macrophages promoted intracellular retention of viral particles in VCC that were no longer connected to the plasma membrane. Finally, we stimulated the rapid release of viral particles from the VCC by subjecting infected macrophages to frustrated phagocytosis. As macrophages spread on IgG-coated glass surfaces, VCC rapidly migrated to the basal membrane and released their viral content in the extracellular medium, which required their association with CD18 and the actin cytoskeleton. These results highlight that VCC trafficking and virus release are intimately linked to the reorganization of the macrophage actin cytoskeleton in response to external physical cues, suggesting that it might be regulated in tissues by the mechanical stress to which these cells are exposed.

Thermoregulatory morphodynamics of honeybee swarm clusters

During reproductive swarming, honeybees clusters of more than 10,000 individuals that hang from structures in the environment (e.g., tree branches) are exposed to diurnal variations in ambient temperature for up to a week. Swarm clusters collectively modulate their morphology in response to these variations (i.e., expanding/contracting in response to heating/cooling) to maintain their internal temperature within a tolerable range and to avoid exhausting their honey stores prematurely. To understand the spatiotemporal aspects of thermoregulatory morphing, we measured the change in size, shape and internal temperature profiles of swarm clusters in response to dynamic temperature ramp perturbations. We see that swarm clusters show a two-fold variation in their volume/density when heated from 15°C to 30°C. However, they do not reach an equilibrium size or shape when held at 30°C for 5 hours, long after the core temperature of the cluster has stabilized. Furthermore, the changes in cluster shape and size are hysteretic, contracting in response to cooling faster than expanding in response to heating. Although the base contact diameter of the cluster increased continuously when the swarm is heated, the change in length of the swarm (base totip) over time is non-monotonic. Consequently, the aspect ratio of the swarm fluctuated continuously even when held at a constant temperature. Taken together, our results quantify the hysteretic and anisotropic morphological responses of swarm clusters to ambient temperature variations while suggesting that both mechanical constraints and heat transfer govern their thermoregulatory morphodynamics.

An Analytical Prediction Model of Balanced and Unbalanced Faults in Doubly-Fed Induction Machines

This paper presents the exact transient solution to the unbalanced and balanced faults in the doubly-fed induction machine (DFIM). Stator currents, rotor currents, and stator fluxes have been validated using simulation and experiment. The work is meant to strengthen and fasten the predictability of large DFIMs in the design stage to comply with mechanical constraints or grid fault issues. Moreover, the analytical approach reduces the computational costs of large-scale stability studies and is especially suited to the initial phase where a plethora design computations must be carried out for the DFIM before it is checked for its transient interaction with the power system. The possibility to dynamically estimate DFIM performance is simplified by original equations derived from first principles. First, a case study of a large 265.50 MVA DFIM is used to verify the proposed "large machine approximation" using simulation, achieving an exact match. Then, laboratory measurements were conducted on a 10.96 kVA and a 1.94 kVA DFIM to validate the transient current peaks predicted in the proposed analytic expressions for two-phase and three-phase faults, respectively.

An Analytical Prediction Model of Balanced and Unbalanced Faults in Doubly-Fed Induction Machines

This paper presents the exact transient solution to the unbalanced and balanced faults in the doubly-fed induction machine (DFIM). Stator currents, rotor currents, and stator fluxes have been validated using simulation and experiment. The work is meant to strengthen and fasten the predictability of large DFIMs in the design stage to comply with mechanical constraints or grid fault issues. Moreover, the analytical approach reduces the computational costs of large-scale stability studies and is especially suited to the initial phase where a plethora design computations must be carried out for the DFIM before it is checked for its transient interaction with the power system. The possibility to dynamically estimate DFIM performance is simplified by original equations derived from first principles. First, a case study of a large 265.50 MVA DFIM is used to verify the proposed "large machine approximation" using simulation, achieving an exact match. Then, laboratory measurements were conducted on a 10.96 kVA and a 1.94 kVA DFIM to validate the transient current peaks predicted in the proposed analytic expressions for two-phase and three-phase faults, respectively.

An attempt to apply the kinematic method of rigid solids in the study of bearing capacity of shallow foundations

In the geotechnical engineering field, shallow foundations are frequently needed to ensure good fieldwork stability. They are also intended to permanently and uniformly transmit all load pressure on the seating floor. However, numerous mechanical constraints, such as bearing capacity of foundations, durability, stability, design of shallow foundations, lead, unfortunately, to a serious realization challenge. Finding an adequate solution presents the main goal and effort of both scholars and professionals. Indeed, the corresponding drawback is observed through the high number of reported damages that occurred in the structure of foundations and the punching failure. The failure mechanisms of shallow foundations, verified in full size or on scale models, show “sliding surfaces” and rigid (solid) blocks, which can be described with the kinematic method of rigid solids. The main objective of this study is the application of the kinematic method of rigid solids in the study of the stability of shallow foundations with respect to punching, the purpose of which is to determine the bearing capacity factors Nc, N γ, and the passive earth pressure coefficient Kp of foundations. In this context, two mechanical models have been proposed with 5 and 7 rigid solids, and a program developed via the MathCAD environment is applied to check the validity of the two previous models. The kinematic method of rigid solids gives results very close and comparable with that of Caquot/Kerisel for the factors of the bearing capacity and passive earth pressure coefficient - the ratio Kp - according to the five- and seven-solid model.

Pitting of malaria parasites in microfluidic devices mimicking spleen interendothelial slits

The spleen is a hematopoietic organ that participates in cellular and humoral immunity. It also serves as a quality control mechanism for removing senescent and/or poorly deformable red blood cells (RBCs) from circulation. Pitting is a specialized process by which the spleen extracts particles, including malaria parasites, from within circulating RBCs during their passage through the interendothelial slits (IES) in the splenic cords. To study this physiological function in vitro, we have developed two microfluidic devices modeling the IES, according to the hypothesis that at a certain range of mechanical stress on the RBC, regulated through both slit size and blood flow, would force it undergo the pitting process without affecting the cell integrity. To prove its functionality in replicating pitting of malaria parasites, we have performed a characterization of P. falciparum-infected RBCs (P.f.-RBCs) after their passage through the devices, determining hemolysis and the proportion of once-infected RBCs (O-iRBCs), defined by the presence of a parasite antigen and absence of DAPI staining of parasite DNA using a flow cytometry-based approach. The passage of P.f.-RBCs through the devices at the physiological flow rate did not affect cell integrity and resulted in an increase of the frequency of O-iRBCs. Both microfluidic device models were capable to replicate the pitting of P.f.-RBCs ex vivo by means of mechanical constraints without cellular involvement, shedding new insights on the role of the spleen in the pathophysiology of malaria.

Sieve tube structural variation in Austrobaileya scandens and its significance for lianescence

Lianas are characterized by large leaf areas and slender stems, a combination of features that require an efficient vascular system. The only extant member of the Austrobaileyaceae, is an endemic twining liana of the tropical Australian forests with well-known xylem hydraulic traits. However, the vascular phloem continuum through aerial organs remains understudied. We analyzed the structure of phloem conduits across leaf veins and stems of A. scandens, combining topological data obtained through light and electron microscopy, with current models of phloem transport. Leaves displayed a low xylem to phloem ratio compared with leaves of other angiosperms, with vascular elements invariant in diameter along the midrib, but tapered across vein hierarchies. Sieve plate pore radii were extremely small: 0.08μm in minor veins, increasing to 0.12μm in the petiole and only to 0.20μm at the base of the stem, tens of meters away. Searcher branches contained tube shaped phloem conduits with a pectin-rich wall, whereas twining stems displayed sieve elements with tangential connections that displayed a greater fraction of the tubes populated with an astonishing number of sieve plates. Hydraulic segmentation of the leaves in Austrobaileyaceae correlate with vesseless leaves that benefit photoassimilate export through volumetric scaling of the sieve tube elements. Yet, compared with canopy dominant trees, the geometrical properties of the sieve tube in twining stems, restrict considerably energy distribution in the sub-canopy layers, potentially favoring the allocation of assimilates toward the elongating branches. Thus, the conductive xylem of twining stems contrasts with a poorly conductive phloem that meets the mechanical constraints of lianescence.

Thermoregulatory morphodynamics of honeybee clusters

During reproductive swarming, honeybees form clusters of more than 10,000 bees that hang from structures in the environment (e.g., tree branches) and are exposed to diurnal variations in ambient temperature for up to one week during the search for a new nesting site. Swarm clusters collectively modulate their morphology in response to these variations (i.e., expanding/contracting in response to heating/cooling) to maintain their internal temperature within a tolerable range and to avoid exhausting their honey stores prematurely. To understand the spatiotemporal aspects of thermoregulatory morphing, we measured the change in size and shape of swarm clusters over time and the internal temperature profiles in response to dynamic temperature ramp perturbations. We found that swarm clusters can achieve a twofold increase/decrease their volume/density when heated from 15°C to 30°C, but they do not reach an equilibrium size or shape when held at 30°C for 5 hours, long after the core temperature of the cluster has stabilized. Furthermore, the changes in cluster shape and size are hysteretic, contracting in response to cooling faster than expanding in response to heating. Although the contact diameter of the cluster increased continuously when the swarm is heated, the change in length of the swarm (base to tip) over time is non-monotonic. Consequently, the aspect ratio of the swarm fluctuated continuously even when held at a constant temperature. Taken together, our results quantify the hysteretic and anisotropic morphological responses of swarm clusters to ambient temperature variations while suggesting that both mechanical constraints and heat transfer govern the thermoregulatory morphing dynamics of swarm clusters.