Abstract. An assay has been developed to quantitatively measure the tension and elasticity of the cytoskeleton in living plant cells. The cell optical displacement assay (CODA) uses a focused laser beam to optically trap and displace transvacuolar and cortical strands through a defined distance within the cell. Results from these experiments provide evidence for the classification of at least two rheologically distinct cytoskeletal assemblies, cortical and transvacuolar, that differ in their tension and response to both signaling molecules and reagents that perturb the cytoskeleton. It is further demonstrated that the tension of the transvacuolar strands can be significantly decreased by the addition of either linoleic acid, 1,2 dioctanoylsn-glycerol, or 1,3 dioctanoylglycerol. These decreases in tension could also be induced by lowering the cytoplasmic pH. In contrast, addition of Ca 2+, Mg 2+, or the ionophore A23187 to the cells caused a considerable increase in the tension of the transvacuolar strands. The data provides evidence that: (a) linoleic acid may be a signaling molecule in plant cells; (b) diacylglycerol functions as a signaling molecule through a protein kinase C-independent pathway mediated by PLA2; and (c) Ca 2+ and pH have regulatory roles for controlling cytoskeleton tension and organization.

A proper understanding of geomechanical behavior of methane hydrate‐bearing sediments is crucial for sustainable future gas production. There are a number of triaxial experiments conducted over synthetic and natural methane hydrate (MH)‐bearing sediments, and several soil constitutive models have been proposed to describe their behavior. However, the generality of a sophisticated model is questioned if it is tested only for a limited number of cases. Furthermore, it is difficult to experimentally determine the associated parameters if their physical meanings and significance are not elucidated. The objective of this paper is to demonstrate that a simple extension of the critical state framework is sufficient to capture the geomechanical behavior of MH‐bearing soils from various sources around the world, while the significance of each parameter is quantified through variance‐based global sensitivity analyses. Our results show that the influence of hydrates can be largely represented by one hydrate‐dependent parameter, pcd′, which controls the expansion of the initial yield surface. This is validated through comparisons with shearing and volumetric response of MH‐bearing soils tested at various institutes under different confining stresses and with varying degrees of hydrate saturation. Our study suggests that the behavior of MH‐bearing soils can be reasonably predicted based on pcd′ and the conventional critical state parameters of the host sediments that can be obtained through typical geotechnical testing procedures.

Fluorescent probe 1, the first inorganic phosphate (Pi) targeted colorimetric and fluorescent probe to detect endogenous Pi in hemichannel-closed cells, has been developed. Probe 1 undergoes a unique Pi induced hydrolytic reaction in DMSO-HEPES (V/V = 9:1) buffered (0.02 M, pH 7.4) solutions that produces a colorimetric change associated with a 62 nm red-shift in the UV-vis absorption maximum and up to a 780-fold enhancement in the fluorescence intensity. The mechanistic proposal that these spectroscopic changes are associated with reaction Pi with 1 to form coumarin gains support from the results of theoretical calculations and mass spectrometry studies. Observations made in fluorescence imaging studies with HeLa cells and C. elegans show that 1 can be employed to monitor Pi production in vivo caused by apyrase-catalyzed ATP hydrolysis. Moreover, probe 1 was utilized to show that apoptosis of hemichannel-closed Sf9 cells is caused by Inx3 promoted dephosphorylation of Akt (RAC serine/threonine-protein kinase), leading to an elevation of the concentration of Pi. Overall, the study has produced the first fluorescent sensor 1 for endogenous inorganic phosphate. Moreover, the utility of 1 for measuring Pi release in vitro has been demonstrated and utilized to elucidate the mechanism of Inx3 action in hemichannel-closed Sf9 cells.

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