Measurement of the Interaction Between Recombinant I-domain from Integrin alpha 2 beta 1 and a Triple Helical Collagen Peptide with the GFOGER Binding Motif Using Molecular Force Spectroscopy.

The role of the collagen-platelet interaction is of crucial importance to the haemostatic response during both injury and pathogenesis of the blood vessel wall. Of particular interest is the high affinity interaction of the platelet transmembrane receptor, alpha 2 beta 1, responsible for firm attachment of platelets to collagen at and around injury sites. We employ single molecule force spectroscopy (SMFS) using the atomic force microscope (AFM) to study the interaction of the I-domain from integrin alpha 2 beta 1 with a synthetic collagen related triple-helical peptide containing the high-affinity integrin-binding GFOGER motif, and a control peptide lacking this sequence, referred to as GPP. By utilising synthetic peptides in this manner we are able to study at the molecular level subtleties that would otherwise be lost when considering cell-to-collagen matrix interactions using ensemble techniques. We demonstrate for the first time the complexity of this interaction as illustrated by the complex multi-peaked force spectra and confirm specificity using control blocking experiments. In addition we observe specific interaction of the GPP peptide sequence with the I-domain. We propose a model to explain these observations.

Normal shock interactions in rectangular channels

Experiments have been conducted to examine the mechanisms behind the coupling between corner separation and centreline separation when holding a normal shock in a rectangular channel. The study has focused on a M ∞ = 1.5 normal shock held in a wind tunnel with a parallel rectangular cross-section. The primary mechanism explaining the link between the corner separation size and the centreline separation appears to be the generation of compression waves which act to smear the adverse pressure gradient imposed upon other parts of the flow. In addition, the origin of the λ-foot leading leg appears to be depended upon the size of the corner separations. Experimental results indicate that the alteration of the λ-region, which occurs in the supersonic portion of the SBLI, is more important than the generation of any blockage in the subsonic region downstream of the shock wave. Copyright © 2012 by H. Babinsky, D.M.F. Burton.

Transducer and base compliance alter the in situ 6 dof force measured from muscle during an isometric contraction in a multi-joint limb.

Although musculoskeletal models are commonly used, validating the muscle actions predicted by such models is often difficult. In situ isometric measurements are a possible solution. The base of the skeleton is immobilized and the endpoint of the limb is rigidly attached to a 6-axis force transducer. Individual muscles are stimulated and the resulting forces and moments recorded. Such analyses generally assume idealized conditions. In this study we have developed an analysis taking into account the compliances due to imperfect fixation of the skeleton, imperfect attachment of the force transducer, and extra degrees of freedom (dof) in the joints that sometimes become necessary in fixed end contractions. We use simulations of the rat hindlimb to illustrate the consequences of such compliances. We show that when the limb is overconstrained, i.e., when there are fewer dof within the limb than are restrained by the skeletal fixation, the compliances of the skeletal fixation and of the transducer attachment can significantly affect measured forces and moments. When the limb dofs and restrained dofs are matched, however, the measured forces and moments are independent of these compliances. We also show that this framework can be used to model limb dofs, so that rather than simply omitting dofs in which a limb does not move (e.g., abduction at the knee), the limited motion of the limb in these dofs can be more realistically modeled as a very low compliance. Finally, we discuss the practical implications of these results to experimental measurements of muscle actions.

The inverse problem in magnetic force microscopy--inferring sample magnetization from MFM images.

Nanomagnetic structures have the potential to surpass silicon's scaling limitations both as elements in hybrid CMOS logic and as novel computational elements. Magnetic force microscopy (MFM) offers a convenient characterization technique for use in the design of such nanomagnetic structures. MFM measures the magnetic field and not the sample's magnetization. As such the question of the uniqueness of the relationship between an external magnetic field and a magnetization distribution is a relevant one. To study this problem we present a simple algorithm which searches for magnetization distributions consistent with an external magnetic field and solutions to the micromagnetic equations' qualitative features. The algorithm is not computationally intensive and is found to be effective for our test cases. On the basis of our results we propose a systematic approach for interpreting MFM measurements.

A canonical normal SBLI flow relevant to external compression inlets

The normal shock wave / boundary layer interaction (normal SBLI) is important to the operation and performance of a supersonic inlet, and the normal SBLI is particularly prominent in external compression inlets. To improve our understanding of such interactions, it is helpful to make use of fundamental flows which capture the main elements of inlets, without resorting to the level of complexity and system integration associated with full-geometry inlets. In this paper, several fundamental fiow-fleld configurations have been considered as possible test cases to represent the normal SBLI aspects found in typical external compression inlets, and it was found that the spillage-diffuser more closely retains the basic flow features of an external compression inlet than the other configurations. In particular, this flow-fleld allows the normal shock Mach number as well as the amount and rate of subsonic diffusion to be all held approximately constant mid independent of the application of flow control. In addition, a survey of several external compression inlets was conducted to quantify the flow and geometric parameters of the spillage-diffuser relevant to actual inlets. The results indicated that such a flow may be especially relevant if the terminal Mach number is about 1.3 to 1.4, the confinement parameter is around 10%, the width around twice or three times the height, and with the area expansion just downstream of the shock on the conservative side of the stall limit for incompressible diffusers. © 2013 by the American Institute of Aeronautics and Astronautics, Inc. All rights reserved.

Flow physics of a normal-hole bled supersonic turbulent boundary layer

An experimental study on normal hole bleed in a supersonic turbulent boundary layer has been conducted. A combination of LDV, Schlieren imagery and oil flow visualization were used to provide a better understanding of the three-dimensional flow field surrounding a supersonic bleed array. Experiments were performed at Mach numbers of 1.8 and 2.5, while previously published results at Mach numbers of 1.3 and 1.5 were also incorporated. The bleed system was capable of removing up to approximately 10% of the incoming boundary layer through a tunnel-spanning array of discrete holes with diameters the same order of magnitude of boundary layer displacement thickness. Inspection of boundary layer profiles downstream of the bleed region indicates that vorticity generated by the discrete holes can have a substantial influence on changes to the boundary layer shape factor and skin friction coefficient, through modification of the lower 20% of the boundary layer. This vorticity was visualized through oil-flow visualization, and LDV measurements, showing the development of two vortices off each bleed hole, and corresponding upwash and downwash regions with far-reaching three dimensional effects. © 2013 by J. M. Oorebeek and H. Babinsky.

The effects of various vortex generator configurations on a normal shock wave/boundary layer interaction

A number of VG configurations have been examined in a inlet relevant fiow-fleld which includes a terminal shock wave and subsequent subsonic diffuser. The flow-fleld was found to be highly sensitive to VG configuration. While the performance of one vane VG configuration was good over a wide range of streamwise positions, another quite similar vane configuration tended to perforin less well-especially when positioned further from the separation-and work is ongoing to determine the reasons behind tliis behavior. In addition, it was found that vane-type VG configurations were appreciably better at reducing separation than their micro-ramp counterparts. When combined with bleed in the centre-span region upstream of the VGs, the performance of vane type VGs was further enhanced and was the best of any configuration. © 2013 by Neil Titchener, Holger Babinsky and Eric Loth.

Canonical normal shock wave/boundary-layer interaction flows relevant to external compression inlets

The normal shock wave/boundary-layer interaction is important to the operation and performance of a supersonic inlet, and the normal shock wave/boundary-layer interaction is particularly prominent in external compression inlets. To improve understanding of such interactions, it is helpful to make use of fundamental flows that capture the main elements of inlets, without resorting to the level of complexity and system integration associated with full-geometry inlets. In this paper, several fundamental flowfield configurations have been considered as possible test cases to represent the normal shock wave/boundary-layer interaction aspects found in typical external compression inlets, and it was found that the spillage diffuser more closely retains the basic flow features of an external compression inlet than the other configurations. In particular, this flowfield allows the normal shock Mach number as well as the amount and rate of subsonic diffusion to all be held approximately constant and independent of the application of flow control. In addition, a survey of several external compression inlets was conducted to quantify the flow and geometric parameters of the spillage diffuser relevant to actual inlets. The results indicated that such a flow may be especially relevant if the terminal Mach number is about 1.3 to 1.4, the confinement parameter is around 10%, and the width is around twice or three times the height. In addition, the area expansion downstream of the shock should be limited to the conservative side of incipient stall based on incompressible diffusers. Copyright © 2013 by the authors.

Generating suspended cell monolayers for mechanobiological studies.

Cell monolayers line most of the surfaces and cavities in the human body. During development and normal physiology, monolayers sustain, detect and generate mechanical stresses, yet little is known about their mechanical properties. We describe a cell culture and mechanical testing protocol for generating freely suspended cell monolayers and examining their mechanical and biological response to uniaxial stretch. Cells are cultured on temporary collagen scaffolds polymerized between two parallel glass capillaries. Once cells form a monolayer covering the collagen and the capillaries, the scaffold is removed with collagenase, leaving the monolayer suspended between the test rods. The suspended monolayers are subjected to stretching by prying the capillaries apart with a micromanipulator. The applied force can be measured for the characterization of monolayer mechanics. Monolayers can be imaged with standard optical microscopy to examine changes in cell morphology and subcellular organization concomitant with stretch. The entire preparation and testing protocol requires 3-4 d.

Functionally different pads on the same foot allow control of attachment: stick insects have load-sensitive "heel" pads for friction and shear-sensitive "toe" pads for adhesion.

Stick insects (Carausius morosus) have two distinct types of attachment pad per leg, tarsal "heel" pads (euplantulae) and a pre-tarsal "toe" pad (arolium). Here we show that these two pad types are specialised for fundamentally different functions. When standing upright, stick insects rested on their proximal euplantulae, while arolia were the only pads in surface contact when hanging upside down. Single-pad force measurements showed that the adhesion of euplantulae was extremely small, but friction forces strongly increased with normal load and coefficients of friction were [Formula: see text] 1. The pre-tarsal arolium, in contrast, generated adhesion that strongly increased with pulling forces, allowing adhesion to be activated and deactivated by shear forces, which can be produced actively, or passively as a result of the insects' sprawled posture. The shear-sensitivity of the arolium was present even when corrected for contact area, and was independent of normal preloads covering nearly an order of magnitude. Attachment of both heel and toe pads is thus activated partly by the forces that arise passively in the situations in which they are used by the insects, ensuring safe attachment. Our results suggest that stick insect euplantulae are specialised "friction pads" that produce traction when pressed against the substrate, while arolia are "true" adhesive pads that stick to the substrate when activated by pulling forces.

Electrostatic force microscopy and potentiometry of realistic nanostructured systems

We investigate the dependency of electrostatic interaction forces on applied potentials in electrostatic force microscopy (EFM) as well as in related local potentiometry techniques such as Kelvin probe microscopy (KPM). The approximated expression of electrostatic interaction between two conductors, usually employed in EFM and KPM, may loose its validity when probe-sample distance is not very small, as often realized when realistic nanostructured systems with complex topography are investigated. In such conditions, electrostatic interaction does not depend solely on the potential difference between probe and sample, but instead it may depend on the bias applied to each conductor. For instance, electrostatic force can change from repulsive to attractive for certain ranges of applied potentials and probe-sample distances, and this fact cannot be accounted for by approximated models. We propose a general capacitance model, even applicable to more than two conductors, considering values of potentials applied to each of the conductors to determine the resulting forces and force gradients, being able to account for the above phenomenon as well as to describe interactions at larger distances. Results from numerical simulations and experiments on metal stripe electrodes and semiconductor nanowires supporting such scenario in typical regimes of EFM investigations are presented, evidencing the importance of a more rigorous modeling for EFM data interpretation. Furthermore, physical meaning of Kelvin potential as used in KPM applications can also be clarified by means of the reported formalism. © 2009 American Institute of Physics.

C1q-like inhibits p53-mediated apoptosis and controls normal hernatopoiesis during zebrafish embryogenesis

Except for the complement C1q, the immunological functions of other C1q family members have remained unclear. Here we describe zebrafish C1q-like, whose transcription and translation display a uniform distribution in early embryos, and are restricted to mid-hind brain and eye in later embryos. In vitro studies showed that C1q-like could inhibit the apoptosis induced by ActD and CHX in EPC cells, through repressing caspase 3/9 activities. Moreover, its physiological roles were studied by morpholino-mediated knockdown in zebrafish embryogenesis. In comparison with control embryos, the C1q-like knockdown embryos display obvious defects in the head and cramofacial development mediated through p53-induced apoptosis, which was confirmed by the in vitro transcribed C1q-like mRNA or p53 MO co-injection. TUNEL assays revealed extensive cell death, and caspase 3/9 activity measurement also revealed about two folds increase in C1q-like morphant embryos, which was inhibited by p53 MO co-injection. Real-time quantitative PCR showed the up-regulation expression of several apoptosis regulators such as p53, mdm2, p21, Box and caspase 3, and down-regulation expression of hbae1 in the C1q-like morphant embryos. Knockdown of C1q-like in zebrafish embryos decreased hemoglobin production and impaired the organization of mesencephalic vein and other brain blood vessels. Interestingly, exposure of zebrafish embryos to UV resulted in an increase in mRNA expression of C1q-like, whereas over-expression of C1q-like was not enough resist to the damage. Furthermore, C1q-like transcription was up-regulated in response to pathogen Aeromonas hydrophila, and embryo survival significantly decreased in the C1q-like morphants after exposure to the bacteria. The data suggested that C1q-like might play an antiapoptotic and protective role in inhibiting p53-dependent and caspase 3/9-mediated apoptosis during embryogenesis, especially in the brain development, and C1q-like should be a novel regulator of cell survival during zebrafish embryogenesis. (c) 2008 Elsevier Inc. All rights reserved.

Active control of normal shock wave/turbulent boundary layer interaction using "Smart" piezoelectric flap actuators

This paper looks at active control of the normal shock wave/turbulent boundary layer interaction (SBLI) using smart flap actuators. The actuators are manufactured by bonding piezoelectric material to an inert substrate to control the bleed/suction rate through a plenum chamber. The cavity provides communication of signals across the shock, allowing rapid thickening of the boundary layer approaching the shock, which splits into a series of weaker shocks forming a lambda shock foot, reducing wave drag. Active control allows optimum control of the interaction, as it would be capable of positioning the control region around the original shock position and control the rate of mass transfer. © 2004 by the American Institute of Aeronautics and Astronautics, Inc. All rights reserved.

Control of normal shock wave / turbulent boundary-layer interaction using streamwise slots

The effect of streamwise slots on the interaction of a normal shock wave / turbulent boundary layer has been investigated experimentally at a Mach number of 1.3. The surface pressure distribution for the controlled interaction was found to be significantly smeared, featuring a distinct plateau. This was due to a change in shock structure from a typical unseparated normal shock wave boundary layer interaction to a large bifurcated Lambda type shock pattern. Boundary layer velocity measurements downstream of the slots revealed a strong spanwise variation of boundary layer properties whereas the modified shock structure was relatively twodimensional. Oil flow visualisation indicated that in the presence of slots the boundary layer surface flow was highly three dimensional and confirmed that the effect of slots was mainly due to suction and blowing similar to that for passive control with uniform surface ventilation. Three hole probe measurements confirmed that the boundary layer was three dimensional and that the slots introduced vortical motion into the flowfield. Results indicate that when applied to an aerofoil, the control device has the potential to reduce wave drag while incurring only small viscous penalties. The introduction of streamwise vorticity may also be beneficial to delay trailing edge separation and the device is thought to be capable of postponing buffet onset. © 2001 by A N Smith.