Biophysical Analysis of Mechanical Signals in Immotile Cilia of Mouse Embryonic Nodes Using Advanced Microscopic Techniques.

Immotile cilia of crown cells at the node of mouse embryos are required for sensing leftward fluid flow that gives rise to the breaking of left-right (L-R) symmetry. The flow-sensing mechanism has long remained elusive, mainly because of difficulties inherent in manipulating and precisely analyzing the cilium. Recent progress in optical microscopy and biophysical analysis has allowed us to study the mechanical signals involving primary cilia. In this study, we used high-resolution imaging with mechanical modeling to assess the membrane tension in a single cilium. Optical tweezers, a technique used to trap sub-micron-sized particles with a highly focused laser beam, allowed us to manipulate individual cilia. Super-resolution microscopy allowed us to discern the precise localization of ciliary proteins. Using this protocol, we provide a method for applying these techniques to cilia in mouse embryonic nodes. This method is widely applicable to the determination of mechanical signals in other cilia.

Pd2 MnGa Metamagnetic Shape Memory Alloy with Small Energy Loss.

Metamagnetic shape memory alloys (MMSMAs) are attractive functional materials owing to their unique properties such as magnetostrain, magnetoresistance, and the magnetocaloric effect caused by magnetic-field-induced transitions. However, the energy loss during the martensitic transformation, that is, the dissipation energy, Edis , is sometimes large for these alloys, which limits their applications. In this paper, a new Pd2 MnGa Heusler-type MMSMA with an extremely small Edis and hysteresis is reported. The microstructures, crystal structures, magnetic properties, martensitic transformations, and magnetic-field-induced strain of aged Pd2 MnGa alloys are investigated. A martensitic transformation from L21 to 10M structures is seen at 127.4 K with a small thermal hysteresis of 1.3 K. The reverse martensitic transformation is induced by applying a magnetic field with a small Edis (= 0.3 J mol-1 only) and a small magnetic-field hysteresis (= 7 kOe) at 120 K. The low values of Edis and the hysteresis may be attributed to good lattice compatibility in the martensitic transformation. A large magnetic-field-induced strain of 0.26% is recorded, indicating the proposed MMSMA's potential as an actuator. The Pd2 MnGa alloy with low values of Edis and hysteresis may enable new possibilities for high-efficiency MMSMAs.

Immotile cilia mechanically sense the direction of fluid flow for left-right determination.

Immotile cilia at the ventral node of mouse embryos are required for sensing leftward fluid flow that breaks left-right symmetry of the body. However, the flow-sensing mechanism has long remained elusive. In this work, we show that immotile cilia at the node undergo asymmetric deformation along the dorsoventral axis in response to the flow. Application of mechanical stimuli to immotile cilia by optical tweezers induced calcium ion transients and degradation of Dand5 messenger RNA (mRNA) in the targeted cells. The Pkd2 channel protein was preferentially localized to the dorsal side of immotile cilia, and calcium ion transients were preferentially induced by mechanical stimuli directed toward the ventral side. Our results uncover the biophysical mechanism by which immotile cilia at the node sense the direction of fluid flow.

Non-Hookean large elastic deformation in bulk crystalline metals.

Crystalline metals can have large theoretical elastic strain limits. However, a macroscopic block of conventional crystalline metals practically suffers a very limited elastic deformation of <0.5% with a linear stress-strain relationship obeying Hooke's law. Here, we report on the experimental observation of a large tensile elastic deformation with an elastic strain of >4.3% in a Cu-based single crystalline alloy at its bulk scale at room temperature. The large macroscopic elastic strain that originates from the reversible lattice strain of a single phase is demonstrated by in situ microstructure and neutron diffraction observations. Furthermore, the elastic reversible deformation, which is nonhysteretic and quasilinear, is associated with a pronounced elastic softening phenomenon. The increase in the stress gives rise to a reduced Young's modulus, unlike the traditional Hooke's law behaviour. The experimental discovery of a non-Hookean large elastic deformation offers the potential for the development of bulk crystalline metals as high-performance mechanical springs or for new applications via "elastic strain engineering."

Flexible and Tough Superelastic Co-Cr Alloys for Biomedical Applications.

The demand for biomaterials has been increasing along with the increase in the population of elderly people worldwide. The mechanical properties and high wear resistance of metallic biomaterials make them well-suited for use as substitutes or as support for damaged hard tissues. However, unless these biomaterials also have a low Young's modulus similar to that of human bones, bone atrophy inevitably occurs. Because a low Young's modulus is typically associated with poor wear resistance, it is difficult to realize a low Young's modulus and high wear resistance simultaneously. Also, the superelastic property of shape-memory alloys makes them suitable for biomedical applications, like vascular stents and guide wires. However, due to the low recoverable strain of conventional biocompatible shape-memory alloys, the demand for a new alloy system is high. The novel body-centered-cubic cobalt-chromium-based alloys in this work provide a solution to both of these problems. The Young's modulus of <001>-oriented single-crystal cobalt-chromium-based alloys is 10-30 GPa, which is similar to that of human bone, and they also demonstrate high wear and corrosion resistance. They also exhibit superelasticity with a huge recoverable strain up to 17.0%. For these reasons, the novel cobalt-chromium-based alloys can be promising candidates for biomedical applications.

Impact of rheological properties on bacterial streamer formation.

Bacterial biofilms, which can be found wherever there is water and a substrate, can cause chronic infections and clogging of industrial flow systems. Despite intensive investigation of the dynamics and rheological properties of biofilms, the impact of their rheological properties on streamer growth remains unknown. We numerically simulated biofilm growth in a pillar-flow and investigated the effects of rheological properties of a filamentous flow-shaped biofilm, called a 'streamer', on its formation by varying the viscoelasticity. The flow-field is assumed to be a Stokes flow and is solved by a boundary element method. A Maxwell model is used for extracellular matrix-mediated streamer growth to express the fluidity of streamer formations. Both high elastic modulus and viscosity are needed for streamer formation, and high viscosity promotes streamer growth at low cell concentrations. Our findings are consistent with experimental observations and can explain the relationship between the cell concentrations and viscosity at which streamers form.

Axial and Nonaxial Migration of Red Blood Cells in a Microtube.

Human red blood cells (RBCs) are subjected to high viscous shear stress, especially during microcirculation, resulting in stable deformed shapes such as parachute or slipper shape. Those unique deformed RBC shapes, accompanied with axial or nonaxial migration, cannot be fully described according to traditional knowledge about lateral movement of deformable spherical particles. Although several experimental and numerical studies have investigated RBC behavior in microchannels with similar diameters as RBCs, the detailed mechanical characteristics of RBC lateral movement-in particular, regarding the relationship between stable deformed shapes, equilibrium radial RBC position, and membrane load-has not yet been fully described. Thus, we numerically investigated the behavior of single RBCs with radii of 4 µm in a circular microchannel with diameters of 15 µm. Flow was assumed to be almost inertialess. The problem was characterized by the capillary number, which is the ratio between fluid viscous force and membrane elastic force. The power (or energy dissipation) associated with membrane deformations was introduced to quantify the state of membrane loads. Simulations were performed with different capillary numbers, viscosity ratios of the internal to external fluids of RBCs, and initial RBC centroid positions. Our numerical results demonstrated that axial or nonaxial migration of RBC depended on the stable deformed RBC shapes, and the equilibrium radial position of the RBC centroid correlated well with energy expenditure associated with membrane deformations.

Cilia and centrosomes: Ultrastructural and mechanical perspectives.

Cilia and centrosomes of eukaryotic cells play important roles in cell movement, fluid transport, extracellular sensing, and chromosome division. The physiological functions of cilia and centrosomes are generated by their dynamics, motions, and forces controlled by the physical, chemical, and biological environments. How an individual cilium achieves its beat pattern and induces fluid flow is governed by its ultrastructure as well as the coordination of associated molecular motors. Thus, a bottom-up understanding of the physiological functions of cilia and centrosomes from the molecular to tissue levels is required. Correlations between the structure and motion can be understood in terms of mechanics. This review first focuses on cilia and centrosomes at the molecular level, introducing their ultrastructure. We then shift to the organelle level and introduce the kinematics and mechanics of cilia and centrosomes. Next, at the tissue level, we introduce nodal ciliary dynamics and nodal flow, which play crucial roles in the organogenetic process of left-right asymmetry. We also introduce respiratory ciliary dynamics and mucous flow, which are critical for protecting the epithelium from drying and exposure to harmful particles and viruses, i.e., respiratory clearance function. Finally, we discuss the future research directions in this field.

Bacterial biomechanics-From individual behaviors to biofilm and the gut flora.

Bacteria inhabit a variety of locations and play important roles in the environment and health. Our understanding of bacterial biomechanics has improved markedly in the last decade and has revealed that biomechanics play a significant role in microbial biology. The obtained knowledge has enabled investigation of complex phenomena, such as biofilm formation and the dynamics of the gut flora. A bottom-up strategy, i.e., from the cellular to the macroscale, facilitates understanding of macroscopic bacterial phenomena. In this Review, we first cover the biomechanics of individual bacteria in the bulk liquid and on surfaces as the base of complex phenomena. The collective behaviors of bacteria in simple environments are next introduced. We then introduce recent advances in biofilm biomechanics, in which adhesion force and the flow environment play crucial roles. We also review transport phenomena in the intestine and the dynamics of the gut flora, focusing on that in zebrafish. Finally, we provide an overview of the future prospects for the field.

Swimming microorganisms acquire optimal efficiency with multiple cilia.

Planktonic microorganisms are ubiquitous in water, and their population dynamics are essential for forecasting the behavior of global aquatic ecosystems. Their population dynamics are strongly affected by these organisms' motility, which is generated by their hair-like organelles, called cilia or flagella. However, because of the complexity of ciliary dynamics, the precise role of ciliary flow in microbial life remains unclear. Here, we have used ciliary hydrodynamics to show that ciliates acquire the optimal propulsion efficiency. We found that ciliary flow highly resists an organism's propulsion and that the swimming velocity rapidly decreases with body size, proportional to the power of minus two. Accordingly, the propulsion efficiency decreases as the cube of body length. By increasing the number of cilia, however, efficiency can be significantly improved, up to 100-fold. We found that there exists an optimal number density of cilia, which provides the maximum propulsion efficiency for all ciliates. The propulsion efficiency in this case decreases inversely proportionally to body length. Our estimated optimal density of cilia corresponds to those of actual microorganisms, including species of ciliates and microalgae, which suggests that now-existing motile ciliates and microalgae have survived by acquiring the optimal propulsion efficiency. These conclusions are helpful for better understanding the ecology of microorganisms, such as the energetic costs and benefits of multicellularity in Volvocaceae, as well as for the optimal design of artificial microswimmers.

Active droplet driven by a collective motion of enclosed microswimmers.

Active fluids containing self-propelled particles are relevant for applications such as self-mixing, micropumping, and targeted drug delivery. With a confined boundary, active fluids can generate bulk flow inside the system, which has the potential to create self-propelled active matter. In this study, we propose that an active droplet is driven by a collective motion of enclosed microswimmers. We show that the droplet migrates via the flow field generated by the enclosed microswimmers; moreover, the locomotion direction depends on the swimming mode of these internal particles. The locomotion mechanism of the droplet can be well explained by interfacial velocity, and the locomotion velocity shows good agreement with the Lighthill-Blake theory. These findings are essential to understand the interplay between the motion of self-propelled particles and the bulk motion response of active matter.

Iron-based superelastic alloys with near-constant critical stress temperature dependence.

Shape memory alloys recover their original shape after deformation, making them useful for a variety of specialized applications. Superelastic behavior begins at the critical stress, which tends to increase with increasing temperature for metal shape memory alloys. Temperature dependence is a common feature that often restricts the use of metal shape memory alloys in applications. We discovered an iron-based superelastic alloy system in which the critical stress can be optimized. Our Fe-Mn-Al-Cr-Ni alloys have a controllable temperature dependence that goes from positive to negative, depending on the chromium content. This phenomenon includes a temperature-invariant stress dependence. This behavior is highly desirable for a range of outer space-based and other applications that involve large temperature fluctuations.

Harnessing random low Reynolds number flow for net migration.

Random noise in low Reynolds number flow has rarely been used to obtain net migration of microscale objects. In this study, we numerically show that net migration of a microscale object can be extracted from random directional fluid forces in Stokes flow, by introducing deformability and inhomogeneous density into the object. We also developed a mathematical framework to describe the deformation-induced migration caused by noise. These results provide a basis for understanding the noise-induced migration of a microswimmer and are useful for harnessing energy from low Reynolds number flow.

The shape effect of flagella is more important than bottom-heaviness on passive gravitactic orientation in Chlamydomonas reinhardtii.

The way the unicellular, biflagellated, green alga Chlamydomonas orients upward has long been discussed in terms of both mechanics and physiology. In this study, we focus on the mechanics, i.e. the 'passive' mechanisms, of gravitaxis. To rotate the body upwards, cellular asymmetry is critical. Chlamydomonas can be depicted as a nearly spherical cell body with two anterior, symmetric flagella. The present study looks at the question of whether the existence of the flagella significantly affects torque generation in upward reorientation. The 'density asymmetry model' assumes that the cell is spherical and bottom-heavy and that the shape and weight of the flagella are negligible, while the 'shape asymmetry model' considers the shape of the flagella. Both our experimental and simulation results revealed a considerable contribution from shape asymmetry to the upward orientation of Chlamydomonas reinhardtii, which was several times larger than that of density asymmetry. From the experimental results, we also quantified the extent of bottom-heaviness, i.e. the distance between the centers of gravity and the figure when the cell body is assumed to be spherical. Our estimation was approximately 30 nm, only one-third of previous assumptions. These findings indicate the importance of the viscous drag of the flagella to the upward orientation, and thus negative gravitaxis, in Chlamydomonas.

Shear-Induced Migration of a Transmembrane Protein within a Vesicle.

Biomembranes feature phospholipid bilayers and serve as the interface between cells or organelles and the extracellular and/or cellular environment. Lipids can move freely throughout the membrane; the lipid bilayer behaves like a fluid. Such fluidity is important in terms of the actions of membrane transport proteins, which often mediate biological functions; membrane protein motion has attracted a great deal of attention. Because the proteins are small, diffusion phenomena are often in play, but flow-induced transport has rarely been addressed. Here, we used a dissipative particle dynamics approach to investigate flow-induced membrane protein transport. We analyzed the drift of a membrane protein located within a vesicle. Under the influence of shear flow, the protein gradually migrated toward the vorticity axis via a random walk, and the probability of retention around the axis was high. To understand the mechanism of protein migration, we varied both shear strength and protein size. Protein migration was induced by the balance between the drag and thermodynamic diffusion forces and could be represented by the Péclet number. These results improve our understanding of flow-induced membrane protein transport.

Swimming of Spermatozoa in a Maxwell Fluid.

It has been suggested that the swimming mechanism used by spermatozoa could be adopted for self-propelled micro-robots in small environments and potentially applied to biomedical engineering. Mammalian sperm cells must swim through a viscoelastic mucus layer to find the egg cell. Thus, understanding how sperm cells swim through viscoelastic liquids is significant not only for physiology, but also for the design of micro-robots. In this paper, we developed a numerical model of a sperm cell in a linear Maxwell fluid based on the boundary element slender-body theory coupling method. The viscoelastic properties were characterized by the Deborah number (De), and we found that, under the prescribed waveform, the swimming speed decayed with the Deborah number in the small-De regime (De < 1.0). The swimming efficiency was independent of the Deborah number, and the decrease in the swimming speed was not significantly affected by the wave pattern.

Passive swimming of a microcapsule in vertical fluid oscillation.

The artificial microswimmer is a cutting-edge technology with applications in drug delivery and micro-total-analysis systems. The flow field around a microswimmer can be regarded as Stokes flow, in which reciprocal body deformation cannot induce migration. In this study, we propose a microcapsule swimmer that undergoes amoeboidlike shape deformations under fluid oscillation conditions. This is a study on the propulsion principle using a capsule with a solid membrane, and one of only a few studies using fluid oscillation. The microswimmer consists of an elastic capsule containing fluid and a rigid sphere. Opposing forces are generated when fluid oscillations are applied, because the densities of the internal fluid and sphere are different. The opposing forces induce nonreciprocal body deformation, which leads to migration of the microswimmer under Stokes flow conditions. Using numerical simulations, we found that the microswimmer propels itself in one of two modes, i.e., stroke swimming or drag swimming. We discuss the feasibility of the proposed microswimmer and show that the most efficient swimmer can migrate tens of micrometers per second. These findings pave the way for future artificial microswimmer designs.

Simulation of the nodal flow of mutant embryos with a small number of cilia: comparison of mechanosensing and vesicle transport hypotheses.

Left-right (L-R) asymmetry in the body plan is determined by nodal flow in vertebrate embryos. Shinohara et al. (Shinohara K et al. 2012 Nat. Commun.3, 622 (doi:10.1038/ncomms1624)) used Dpcd and Rfx3 mutant mouse embryos and showed that only a few cilia were sufficient to achieve L-R asymmetry. However, the mechanism underlying the breaking of symmetry by such weak ciliary flow is unclear. Flow-mediated signals associated with the L-R asymmetric organogenesis have not been clarified, and two different hypotheses-vesicle transport and mechanosensing-are now debated in the research field of developmental biology. In this study, we developed a computational model of the node system reported by Shinohara et al. and examined the feasibilities of the two hypotheses with a small number of cilia. With the small number of rotating cilia, flow was induced locally and global strong flow was not observed in the node. Particles were then effectively transported only when they were close to the cilia, and particle transport was strongly dependent on the ciliary positions. Although the maximum wall shear rate was also influenced by ciliary position, the mean wall shear rate at the perinodal wall increased monotonically with the number of cilia. We also investigated the membrane tension of immotile cilia, which is relevant to the regulation of mechanotransduction. The results indicated that tension of about 0.1 µN m-1 was exerted at the base even when the fluid shear rate was applied at about 0.1 s-1. The area of high tension was also localized at the upstream side, and negative tension appeared at the downstream side. Such localization may be useful to sense the flow direction at the periphery, as time-averaged anticlockwise circulation was induced in the node by rotation of a few cilia. Our numerical results support the mechanosensing hypothesis, and we expect that our study will stimulate further experimental investigations of mechanotransduction in the near future.

Asymmetry in cilia configuration induces hydrodynamic phase locking.

To gain insight into the nature of biological synchronization at the microscopic scale, we here investigate the hydrodynamic synchronization between conically rotating objects termed nodal cilia. A mechanical model of three rotating cilia is proposed with consideration of variation in their shapes and geometrical arrangement. We conduct numerical estimations of both near-field and far-field hydrodynamic interactions, and we apply a conventional averaging method for weakly coupled oscillators. In the nonidentical case, the three cilia showed stable locked-phase differences around ±π/2. However, such phase locking also occurred with three identical cilia when allocated in a triangle except for the equilateral triangle. The effects of inhomogeneity in cilia shapes and geometrical arrangement on such asymmetric interaction is discussed to understand the role of biological variation in synchronization via hydrodynamic interactions.

Ultra-large single crystals by abnormal grain growth.

Producing a single crystal is expensive because of low mass productivity. Therefore, many metallic materials are being used in polycrystalline form, even though material properties are superior in a single crystal. Here we show that an extraordinarily large Cu-Al-Mn single crystal can be obtained by abnormal grain growth (AGG) induced by simple heat treatment with high mass productivity. In AGG, the sub-boundary energy introduced by cyclic heat treatment (CHT) is dominant in the driving pressure, and the grain boundary migration rate is accelerated by repeating the low-temperature CHT due to the increase of the sub-boundary energy. With such treatment, fabrication of single crystal bars 70 cm in length is achieved. This result ensures that the range of applications of shape memory alloys will spread beyond small-sized devices to large-scale components and may enable new applications of single crystals in other metallic and ceramics materials having similar microstructural features.Growing large single crystals cheaply and reliably for structural applications remains challenging. Here, the authors combine accelerated abnormal grain growth and cyclic heat treatments to grow a superelastic shape memory alloy single crystal to 70 cm.