Heterogeneous flow kinematics of cellulose nanofibril suspensions under shear.

The rheology of NFC suspensions that exhibited different microstructures and colloidal stability, namely TEMPO and enzymatic NFC suspensions, was investigated at the macro and mesoscales using a transparent Couette rheometer combined with optical observations and ultrasonic speckle velocimetry (USV). Both NFC suspensions showed a complex rheology, which was typical of yield stress, non-linear and thixotropic fluids. Hysteresis loops and erratic evolutions of the macroscale shear stress were also observed, thereby suggesting important mesostructural changes and/or inhomogeneous flow conditions. The in situ optical observations revealed drastic mesostructural changes for the enzymatic NFC suspensions, whereas the TEMPO NFC suspensions did not exhibit mesoscale heterogeneities. However, for both suspensions, USV measurements showed that the flow was heterogeneous and exhibited complex situations with the coexistence of multiple flow bands, wall slippage and possibly multidimensional effects. Using USV measurements, we also showed that the fluidization of these suspensions could presumably be attributed to a progressive and spatially heterogeneous transition from a solid-like to a liquid-like behavior. As the shear rate was increased, the multiple coexisting shear bands progressively enlarged and nearly completely spanned over the rheometer gap, whereas the plug-like flow bands were eroded.

Flow-induced structures versus flow instabilities.

The Taylor-Couette flow of a dilute micellar system known to generate shear-induced structures is investigated through simultaneous rheometry and ultrasonic imaging. We show that flow instabilities must be taken into account since both Reynolds and Weissenberg numbers may be large. Before nucleation of shear-induced structures, the flow can be inertially unstable, but once shear-induced structures are nucleated, the kinematics of the flow become chaotic, in a pattern reminiscent of the elastically dominated turbulence known in dilute polymer solutions. We outline a general framework for the interplay between flow instabilities and flow-induced structures.

Polarity proteins in migration and invasion.

Cancer is the result of the deregulation of cell proliferation and cell migration. In advanced tumors, cells invade the surrounding tissue and eventually form metastases. This is particularly evident in carcinomas in which epithelial cells have undergone epithelial-mesenchymal transition. Increased cell migration often correlates with a weakening of intercellular interactions. Junctions between neighboring epithelial cells are required to establish and maintain baso-apical polarity, suggesting that not only loss of cell-cell adhesion but also alteration of cell polarity is involved during invasion. Accordingly, perturbation of cell polarity is an important hallmark of advanced invasive tumors. Cell polarity is also essential for cell migration. Indeed, a front-rear polarity axis has first to be generated to allow a cell to migrate. Because cells migrate during invasion, cell polarity is not completely lost. Instead, polarity is modified. From a nonmigrating baso-apically polarized epithelial phenotype, cells acquire a polarized migrating mesenchymal phenotype. The aim of this review is to highlight the molecular relationship between the control of cell polarity and the regulation of cell motility during oncogenesis.

Ultrasound velocimetry in a shear-thickening wormlike micellar solution: evidence for the coexistence of radial and vorticity shear bands.

We carried out pointwise local velocity measurements on 40 mM cetylpyridinium chloride-sodium salicylate (CPyCl-NaSal) wormlike micellar solution using high-frequency ultrasound velocimetry in a Couette shear cell. The studied wormlike solution exhibits Newtonian, shear-thinning and shear-thickening rheological behavior in a stress-controlled environment. Previous rheology, flow visualization and small-angle light/neutron scattering experiments in the shear-thickening regime of this system showed the presence of stress-driven alternating transparent and turbid rings or vorticity bands along the axis of the Couette geometry. Through local velocity measurements we observe a homogeneous flow inside the 1mm gap of the Couette cell in the shear-thinning (stress-plateau) region. Only when the solution is sheared beyond the critical shear stress (shear-thickening regime) in a stress-controlled experiment, we observe inhomogeneous flow characterized by radial or velocity gradient shear bands with a highly sheared band near the rotor and a weakly sheared band near the stator of the Couette geometry. Furthermore, fast measurements performed in the shear-thickening regime to capture the temporal evolution of local velocities indicate coexistence of both radial and vorticity shear bands. However the same measurements carried out in shear rate controlled mode of the rheometer do not show such rheological complexity.

Local velocity measurements in heterogeneous and time-dependent flows of a micellar solution.

We present and discuss the results of pointwise velocity measurements performed on a viscoelastic micellar solution made of cetyltrimethylammonium bromide and sodium salicylate in water, respectively, at the concentrations of 50 and 100 mmol. The sample is contained in a Couette device and subjected to flow in the strain controlled mode. This particular solution shows shear banding and, in a narrow range of shear rates at the right end of the stress plateau, apparent shear thickening occurs. Time-dependent recordings of the shear stress in this range reveal that the flow has become unstable and that large sustained oscillations of the shear stress and of the first normal stresses difference emerge and grow in the flow. Local pointwise velocity measurements clearly reveal a velocity profile typical of shear banding when the imposed shear rate belongs to the plateau, but also important wall slip in the entire range of velocity gradients investigated. In the oscillations regime, the velocity is recorded as a function of time at a fixed point close to the rotor of the Couette device. The time-dependent velocity profile reveals random fluctuations but, from time to time, sharp decreases much larger than the standard deviation are observed. An attempt is made to correlate these strong variations with the stress oscillations and a correlation coefficient r is computed. However, the small value found for the coefficient r does not allow us to draw a final conclusion as concerns the correlation between stress oscillations and velocity fast decreases.

A spatio-temporal study of rheo-oscillations in a sheared lamellar phase using ultrasound.

We present an experimental study of the flow dynamics of a lamellar phase sheared in the Couette geometry. High-frequency ultrasonic pulses at 36 MHz are used to measure time-resolved velocity profiles. Oscillations of the viscosity occur in the vicinity of a shear-induced transition between a high-viscosity disordered fluid and a low-viscosity ordered fluid. The phase coexistence shows up as shear bands on the velocity profiles. We show that the dynamics of the rheological data result from two different processes: (i) fluctuations of slip velocities at the two walls and (ii) flow dynamics in the bulk of the lamellar phase. The bulk dynamics are shown to be related to the displacement of the interface between the two differently sheared regions in the gap of the Couette cell. Two different dynamical regimes are investigated under applied shear stress: one of small amplitude oscillations of the viscosity delta eta/eta approximately equal to 3%) and one of large oscillations (delta eta/eta approximately equal to 25%). A phenomenological model is proposed that may account for the observed spatio-temporal dynamics.

Towards local rheology of emulsions under Couette flow using Dynamic Light Scattering.

We present local velocity measurements in emulsions under shear using heterodyne Dynamic Light Scattering. Two emulsions are studied: a dilute system of volume fraction phi = 20% and a concentrated system with phi = 75%. Velocity profiles in both systems clearly show the presence of wall slip. We investigate the evolution of slip velocities as a function of shear stress and discuss the validity of the corrections for wall slip classically used in rheology. Focussing on the bulk flow, we show that the dilute system is Newtonian and that the concentrated emulsion is shear-thinning. In the latter case, the curvature of the velocity profiles is compatible with a shear-thinning exponent of 0.4 consistent with global rheological data. However, even if individual profiles can be accounted for by a power law fluid (with or without a yield stress), we could not find a fixed set of parameters that would fit the whole range of applied shear rates. Our data, thus, raise the question of the definition of a global flow curve for such a concentrated system. These results show that local measurements are a crucial complement to standard rheological tools. They are discussed in the light of recent works on soft glassy materials.

Integrin-mediated activation of Cdc42 controls cell polarity in migrating astrocytes through PKCzeta.

We describe here a signal transduction pathway controlling the establishment of mammalian cell polarity. Scratching a confluent monolayer of primary rat astrocytes leads to polarization of cells at the leading edge. The microtubule organizing center, the microtubule cytoskeleton, and the Golgi reorganize to face the new free space, and directed cell protrusion and migration specifically occur perpendicularly to the scratch. We show here that the interaction of integrins with extracellular matrix at the newly formed cell front leads to the activation and polarized recruitment of Cdc42, which in turn recruits and activates a cytoplasmic mPar6/PKCzeta complex. Localized PKCzeta activity, acting through the microtubule motor protein dynein, is required for all aspects of induced polarity in these cells.

Scattering of sound by a vorticity filament: an experimental and numerical investigation.

A vorticity filament is investigated experimentally using the transmission of an ultrasonic wave through the flow. The analysis of the wave-front distortion provides noninvasive measurements of the vortex circulation and size. The latter is estimated by analytical calculations of the scattering of a plane wave by a vorticity filament. The case of a cylindrical wave incident on a vortex leads to similar experimental results which are successfully compared to a parabolic equation simulation. Finally, a finite-difference code based on linear acoustics is presented, in order to investigate the structure of the scattered wave numerically.

ICAM-1-coupled cytoskeletal rearrangements and transendothelial lymphocyte migration involve intracellular calcium signaling in brain endothelial cell lines.

Endothelium of the cerebral blood vessels, which constitutes the blood-brain barrier, controls adhesion and trafficking of leukocytes into the brain. Investigating signaling pathways triggered by the engagement of adhesion molecules expressed on brain endothelial cells using two rat brain endothelial cell lines (RBE4 and GP8), we report in this paper that ICAM-1 cross-linking induces a sustained tyrosine phosphorylation of the phosphatidylinositol-phospholipase C (PLC)gamma1, with a concomitant increase in both inositol phosphate production and intracellular calcium concentration. Our results suggest that PLC are responsible, via a calcium- and protein kinase C (PKC)-dependent pathway, for p60Src activation and tyrosine phosphorylation of the p60Src substrate, cortactin. PKCs are also required for tyrosine phosphorylation of the cytoskeleton-associated proteins, focal adhesion kinase and paxillin, but not for ICAM-1-coupled p130Cas phosphorylation. PKC's activation is also necessary for stress fiber formation induced by ICAM-1 cross-linking. Finally, cell pretreatment with intracellular calcium chelator or PKC inhibitors significantly diminishes transmonolayer migration of activated T lymphocytes, without affecting their adhesion to brain endothelial cells. In summary, our data demonstrate that ICAM-1 cross-linking induces calcium signaling which, via PKCs, mediates phosphorylation of actin-associated proteins and cytoskeletal rearrangement in brain endothelial cell lines. Our results also indicate that these calcium-mediated intracellular events are essential for lymphocyte migration through the blood-brain barrier.

ICAM-1-coupled signaling pathways in astrocytes converge to cyclic AMP response element-binding protein phosphorylation and TNF-alpha secretion.

In the CNS, astrocytes play a key role in immunological and inflammatory responses through ICAM-1 expression, cytokine secretion (including TNF-alpha), and regulation of blood-brain barrier permeability. Because ICAM-1 transduces intracellular signals in lymphocytes and endothelial cells, we investigated in the present study ICAM-1-coupled signaling pathways in astrocytes. Using rat astrocytes in culture, we report that ICAM-1 binding by specific Abs induces TNF-alpha secretion together with phosphorylation of the transcription factor cAMP response element-binding protein. We show that ICAM-1 binding induces cAMP accumulation and activation of the mitogen-activated protein kinase extracellular signal-regulated kinase. Both pathways are responsible for cAMP response element-binding protein phosphorylation and TNF-alpha secretion. Moreover, these responses are partially dependent protein kinase C, which acts indirectly, as a common activator of cAMP/protein kinase A and extracellular signal-regulated kinase pathways. These results constitute the first evidence of ICAM-1 coupling to intracellular signaling pathways in glial cells and demonstrate the convergence of these pathways onto transcription factor regulation and TNF-alpha secretion. They strongly suggest that ICAM-1-dependent cellular adhesion to astrocytes could contribute to the inflammatory processes observed during leukocyte infiltration in the CNS.