A Synthetic Minimal Beating Axoneme.

Cilia and flagella are beating rod-like organelles that enable the directional movement of microorganisms in fluids and fluid transport along the surface of biological organisms or inside organs. The molecular motor axonemal dynein drives their beating by interacting with microtubules. Constructing synthetic beating systems with axonemal dynein capable of mimicking ciliary beating still represents a major challenge. Here, the bottom-up engineering of a sustained beating synthoneme consisting of a pair of microtubules connected by a series of periodic arrays of approximately eight axonemal dyneins is reported. A model leads to the understanding of the motion through the cooperative, cyclic association-dissociation of the molecular motor from the microtubules. The synthoneme represents a bottom-up self-organized bio-molecular machine at the nanoscale with cilia-like properties.

Aeroacoustic characteristics of owl-inspired blade designs in a mixed flow fan: effects of leading- and trailing-edge serrations.

There is an increasing need in industry for noise reduction in fans. Inspired by owls' silent flight, we propose four owl-inspired blade designs for a mixed-flow fan to examine whether leading-edge (LE) and/or trailing-edge (TE) serrations can resolve the tradeoff between sound suppression and aerodynamic performance. We investigate the blades' aeroacoustic characteristics through various experimental methods and large-eddy simulation (LES)-based numerical analyses. Experimental results suggest that 'slotted', simply-fabricated LE serrations can achieve a lowering of the noise level while sustaining the aerodynamic performance of the fan, whereas TE serrations fail. In addition, the inclination angle can improve LE serration performance in aeroacoustic and aerodynamic performance with a reduction in the specific noise level by around 1.4 dB. LES results and noise spectral analysis indicate that the LE serrations can suppress flow separation, reducing the broadband noise at low-to-middle frequencies (40-4k Hz). This passive-flow-control mechanism, likely due to local higher incidence angles associated with LE serrations, is capable of alleviating the intensive pressure gradient while suppressing wall-pressure fluctuations over the LE region, hence weakening the Kelvin-Helmholtz instability. The tonal noise also shows a marked reduction at the highest peak frequency associated with fan-vane interaction. Moreover, we find that the high-frequency noise by-product radiates mainly from the LE serrations andsurroundings, due to the small eddies broken up when the vortical flows pass through the LE serrations. Our results demonstrate that the biomimetic design of the LE serrations can facilitate the break-up of LE vortices passively and effectively without negatively impacting aerodynamic performance, which can be utilized as an effective device to improve the aeroacoustic performance of fan blades.

Evaluation of intercellular cross-linking abilities correlated with cytotoxicities of bispecific antibodies with domain rearrangements using AFM force-sensing.

Bispecific antibodies (bsAbs) are a promising engineered antibody format; thus, technologies for the fabrication and evaluation of functional bsAbs are attracting increasing attention. Here, based on atomic force microscopy (AFM) force-sensing integrated with a metal cup-attached AFM chip (cup-chip) to ensure efficient capture of a target cell on a cantilever, we established a novel method for measuring cross-linking ability that is correlated with the cytotoxicities of bsAbs targeting two cells. We previously reported that domain rearrangements of bsAbs affected their cytotoxicities; however, no differences in cross-linking ability for soluble antigens were observed by surface plasmon resonance. We predicted that there would be differences in molecular configurations to avoid steric hindrance in the cross-linking of the two whole target cells. A picked-up T cell lymphoma cell on the cantilever using a cup-chip was moved to approach a cancer cell adhered to a dish, and force-curve measurements were performed. The resulting forces mediated by the cross-linking of bsAbs with different domain orders were well-correlated with their cytotoxicities. The AFM force-sensing method established herein may reflect steric hindrance of intercellular cross-linking, and thus has the potential to evaluate the net function of bsAbs and contribute to the generation of functional bsAbs.

Influence of Nivolumab for Intercellular Adhesion Force between a T Cell and a Cancer Cell Evaluated by AFM Force Spectroscopy.

The influence of nivolumab on intercellular adhesion forces between T cells and cancer cells was evaluated quantitatively using atomic force microscopy (AFM). Two model T cells, one expressing high levels of programmed cell death protein 1 (PD-1) (PD-1high Jurkat) and the other with low PD-1 expression levels (PD-1low Jurkat), were analyzed. In addition, two model cancer cells, one expressing programmed death-ligand 1 (PD-L1) on the cell surface (PC-9, PD-L1+) and the other without PD-L1 (MCF-7, PD-L1-), were also used. A T cell was attached to the apex of the AFM cantilever using a cup-attached AFM chip, and the intercellular adhesion forces were measured. Although PD-1high T cells adhered strongly to PD-L1+ cancer cells, the adhesion force was smaller than that with PD-L1- cancer cells. After the treatment of PD-1high T cells with nivolumab, the adhesion force with PD-L1+ cancer cells increased to a similar level as with PD-L1- cancer cells. These results can be explained by nivolumab influencing the upregulation of the adhesion ability of PD-1high T cells with PD-L1+ cancer cells. These results were obtained by measuring intercellular adhesion forces quantitatively, indicating the usefulness of single-cell AFM analysis.

Force-Generating Mechanism of Axonemal Dynein in Solo and Ensemble.

In eukaryotic cilia and flagella, various types of axonemal dyneins orchestrate their distinct functions to generate oscillatory bending of axonemes. The force-generating mechanism of dyneins has recently been well elucidated, mainly in cytoplasmic dyneins, thanks to progress in single-molecule measurements, X-ray crystallography, and advanced electron microscopy. These techniques have shed light on several important questions concerning what conformational changes accompany ATP hydrolysis and whether multiple motor domains are coordinated in the movements of dynein. However, due to the lack of a proper expression system for axonemal dyneins, no atomic coordinates of the entire motor domain of axonemal dynein have been reported. Therefore, a substantial amount of knowledge on the molecular architecture of axonemal dynein has been derived from electron microscopic observations on dynein arms in axonemes or on isolated axonemal dynein molecules. This review describes our current knowledge and perspectives of the force-generating mechanism of axonemal dyneins in solo and in ensemble.

Mechanical Property Changes in Breast Cancer Cells Induced by Stimulation with Macrophage Secretions in Vitro.

The contribution of secretions from tumor-associated macrophage (TAM)-like cells to the stimulation of mechanical property changes in murine breast cancer cells was studied using an in vitro model system. A murine breast cancer cell line (FP10SC2) was stimulated by adding macrophage (J774.2) cultivation medium containing stimulation molecules secreted from the macrophages, and changes in mechanical properties were compared before and after stimulation. As a result, cell elasticity decreased, degradation ability of the extracellular matrix increased, and the expression of plakoglobin was upregulated. These results indicate that cancer cell malignancy is upregulated by this stimulation. Moreover, changes in intercellular adhesion strengths between pairs of cancer cells were measured before and after stimulation using atomic force microscopy (AFM). The maximum force required to separate cells was increased by stimulation with the secreted factors. These results indicate the possibility that TAMs cause changes in the mechanical properties of cancer cells in tumor microenvironments, and in vitro measurements of mechanical property changes in cancer cells will be useful to study interactions between cells in tumor microenvironments.

Quantitative Measurements of Intercellular Adhesion Strengths between Cancer Cells with Different Malignancies Using Atomic Force Microscopy.

Intercellular adhesion strengths between two kinds of murine breast cancer cells with different malignancies were measured quantitatively using a metal cup-attached chip with atomic force microscopy (AFM). The cup-attached chip was used to approach a cell, pick it up, and then approach another cell, and the adhesion strengths were measured according to the contact time of the cells between 0 to 60 s. Separation work was used as a parameter for quantitative comparisons of the strengths. As a result, the work of a highly metastatic cancer cell (FP10SC2) was greater than a low metastatic cancer cell (4T1-LM) throughout all contact times examined. Adhesion was analyzed from a point of a view of binding kinetics of receptors on cells, and two possibilities were found: one was the number of cell adhesive receptors increased, and the other was the work to separate single molecular binding increased with increasing cancer cell malignancy. These results indicated quantitative measurements of intercellular adhesion strengths using AFM yielded information to understand the mechanism of the cancer progression from a new perspective.

Oscillation collapse in coupled quantum van der Pol oscillators.

The classical self-oscillations can collapse merely due to their mutual couplings. We investigate this oscillation collapse in quantum van der Pol oscillators. For a pair of quantum oscillators, the steady-state mean phonon number is shown to be lower than in the corresponding classical model with a Gaussian white noise that mimics quantum noise. We further show within the mean-field theory that a number of globally coupled oscillators undergo a transition from the synchronized periodic motion to the collective oscillation collapse. A quantum many-body simulation suggests that the increase in the number of oscillators leads to a lower steady-state mean phonon number, bounded below by the mean-field result.