Membrane waves driven by actin and Myosin

We present a model which couples the membrane with the protrusive forces of actin polymerization and contractile forces of molecular motors, such as myosin. The actin polymerization at the membrane is activated by freely diffusing membrane proteins that have a spontaneous curvature. Molecular motors are recruited to the polymerizing actin filaments, from a constant reservoir, and produce a contractile force. All the forces and variables are treated in the linear limit. Our results show that for convex membrane proteins the myosin activity gives rise to robust transverse membrane waves, similar to those observed on different cells.     

Direct Visualization of Vesicle Maturation and Plasma Membrane Protein Trafficking

Internalization and intracellular trafficking of membrane proteins are now recognized as essential mechanisms that contribute to a number of cellular processes. Current methods lack the ability to specifically label the plasma membrane of a live cell, follow internalization of labeled membrane molecules, and conclusively differentiate newly formed membrane-derived vesicles from pre-existing endocytic or secretory structures in the cytoplasm. Here, we detail a visualization method for surface biotinylation of plasma membrane-derived vesicles that allows us to follow their progress from membrane to cytosol at specific time points. Using the transmembrane receptor RET as a model, we demonstrate how this method can be applied to identify plasma membrane-derived vesicle maturation, determine RET’s presence within these structures, and monitor RET’s recycling to the cell surface. This method improves on static and less discriminatory methods, providing a tool for analysis of real-time vesicle trafficking that is applicable to many systems.     

Correlated light and electron microscopy observations of the uterine epithelial cell actin cytoskeleton using fluorescently labeled resin-embedded sections

In order to perform correlative light and electron microscopy (CLEM) more precisely, we have modified existing specimen preparation protocols allowing fluorescence retention within embedded and sectioned tissue, facilitating direct observation across length scales. We detail a protocol which provides a precise correlation accuracy using accessible techniques in biological specimen preparation. By combining a pre-embedding uranyl acetate staining step with the progressive lowering of temperature (PLT) technique, a methacrylate embedded tissue specimen is ultrathin sectioned and mounted onto a TEM finder grid for immediate viewing in the confocal and electron microscope. In this study, the protocol is applied to rat uterine epithelial cells in vivo during early pregnancy. Correlative overlay data was used to track changes in filamentous actin that occurs in these cells from fertilization (Day 1) to implantation on Day 6 as part of the plasma membrane transformation, a process essential in the development of uterine receptivity in the rat. CLEM confirmed that the actin cytoskeleton is disrupted as apical microvilli are progressively lost toward implantation, and revealed the thick and continuous terminal web is replaced by a thinner and irregular actin band, with individually distinguishable filaments connecting actin meshworks which correspond with remaining plasma membrane protrusions.     

The rectum of Oxaea flavescens (Andrenidae) has a specialized structure among bees

The rectum of the bee Oxaea flavescens contains six hollow rectal papillae, in contrast to rectal pads found in others Hymenoptera. The rectal epithelium is formed by cubical cells with structural specializations for fluid transport, such as apical plasma membrane invaginations, endocytic vesicles, scalariform junctions, wide extracellular space and double-membrane vesicles. Globular-shaped cells form the rectal papillae with surface plasma membrane invaginations that vary from small depressions to very large sinuses. The cell-cell contact is narrow, exhibiting extensive regions with scalariform junction, while cytoplasmic free ribosome-like material is prominent. These ultrastructural features are discussed in relation to their role in excretion in this bee.     

Cytoskeletal turnover and Myosin contractility drive cell autonomous oscillations in a model of Drosophila Dorsal Closure

Oscillatory behaviour in force-generating systems is a pervasive phenomenon in cell biology. In this work, we investigate how oscillations in the actomyosin cytoskeleton drive cell shape changes during the process of Dorsal Closure (DC), a morphogenetic event in Drosophila embryo development whereby epidermal continuity is generated through the pulsatile apical area reduction of cells constituting the amnioserosa (AS) tissue. We present a theoretical model of AS cell dynamics by which the oscillatory behaviour arises due to a coupling between active myosin-driven forces, actin turnover and cell deformation. Oscillations in our model are cell-autonomous and are modulated by neighbour coupling, and our model accurately reproduces the oscillatory dynamics of AS cells and their amplitude and frequency evolution. A key prediction arising from our model is that the rate of actin turnover and Myosin contractile force must increase during DC in order to reproduce the decrease in amplitude and period of cell area oscillations observed in vivo. This prediction opens up new ways to think about the molecular underpinnings of AS cell oscillations and their link to net tissue contraction and suggests the form of future experimental measurements.     

Histology and ultrastructure of pericardial cells of Scaptotrigona postica Latreille (Hymenoptera, Apidae) in workers and queens of different ages

The paper presents a study of the pericardial cells of Scaptotrigona postica an eusocial Brazilian stingless bee. Light and electron microscopy was used in a comparative study on workers and queens of different ages, exerting different functions in the colony. The pericardial cells are found only in the pericardial sinus, mainly in groups around the dorsal vessel. Each cell is enclosed by the basal membrane and its peripheral region is characterized by folds of the plasma membrane, which form canals and loops. The points where the plasma membrane folds is frequently closed by diaphragms, that along with the basal lamina form a barrier to substances from hemolymph. Along the membrane limiting the canals and loops, an intense endocytic activity through coated vesicles takes place indicating a selective absorption of hemolymph components. In older individuals, workers or queens, the cells exhibit larger quantities of cytoplasm inclusions, heterogeneous vacuoles containing the final products of intracellular digestion, and autophagic vacuoles with concentric membranous structures. The pericardial cells general morphology is in accordance with the role in processing metabolites captured from hemolymph and storage of indigested residues.     

Intrinsic Fluorescence of Actin

In this work actin is used to illustrate connection of protein fluorescence characteristics with its structure. On one hand, it has been demonstrated what kind of information about the contribution of each tryptophan residues to the bulk fluorescence spectrum can be obtained from the special analysis of protein three-dimensional structure. On the other hand, potentials of intrinsic fluorescence for elucidation of proteins structure, dynamics and processes of folding-unfolding are shown. In particular, using this method a new essentially unfolded kinetic intermediate state of actin was detected and characterized, and the place of inactivated actin and its kinetic predecessor in the process of folding-unfolding was determined. It has been revealed that inactivated actin is not intermediate state between the native and completely unfolded states, as it has been accepted before, but a result of protein misfolding. On the basis of the obtained data a new model of actin folding-unfolding pathway has been proposed.     

Dynamics of cell membranes and the underlying cytoskeletons observed by noninterferometric widefield optical profilometry and fluorescence microscopy

Combining the noninterferometric wide-field optical profilometry technique with fluorescence microscopy, we observe the membrane activities of a living cell as well as the structures of its cytoskeletons. The membrane ripples of a lamellipodium are related to similar structures of the underlying actin filaments. However, we find the ripples appear prior to and disappear later than the corresponding actin filament structures, which supports the elastic Brownian ratchet model of cell motility. In addition, we measure the three-dimensional movement of a fibronectin-coated latex bead on the membrane. The bead motion is determined by the movement and branching of the actin molecules on the filament, as well as by the displacement of the filament itself.     

Formation of complex structures in dusty plasmas under temperature gradients

Thermophoretic effects on dust structures under temperature gradients in glow and radio-frequency discharge plasmas are studied experimentally. The geometry of dust structures consisting of micrometer-sized polydisperse grains depends on heat release in the plasma. Thermophoretic forces associated with heat release can control the formation of dust structures of different geometries. A theoretical model is proposed to describe dust separation with respect to grain size caused by the effects of radial electrostatic and thermophoretic forces. The glow discharge currents under critical conditions for grain separation predicted by the model agree with those observed experimentally.     

常气压辉光放电等离子体对边界层流动的影响

基于Shyy提出的常气压下均匀辉光放电等离子体与空气干扰的物理模型,通过求解电位势方程得到电场分布及作用于流体上的电场力.以NACA0015翼型低速绕流为对象,通过数值求解考虑等离子体作用的流体运动控制方程,研究等离子体位置和个数控制对翼型绕流分离的影响.位于分离点上游的等离子体能够有效地抑制流动分离,而在分离区的等离子体对流动影响很弱,这一结论同实验观察一致,并给出等离子体对翼型壁面压力和气动力影响的规律.     

Inductive interaction of rapidly rotating conductive bodies with a magnetized plasma

This paper discusses the magnetohydrodynamic flow of a supersonic plasma flux around a thin conductive body. The specific features of the inductive interaction of the body with the plasma and the process by which plasma waves are generated when the body rotates are investigated. The structure of the magnetic-field perturbations and the distribution of the plasma currents that result from the inductive interaction are studied. Expressions are obtained for the forces that act on a plate and the torques produced by these forces. A simple model is used to take into account the kinetic effects associated with the finiteness of the absorption and emission currents that transport charge from the plate to the plasma and back. A surface-potential distribution is found that can substantially accelerate particles in the neighborhood of the body. Zh. éksp. Teor. Fiz. 115, 542–563 (February 1999)     

Force balance and membrane shedding at the red-blood-cell surface

During the aging of the red-blood cell, or under conditions of extreme echinocytosis, membrane is shed from the cell plasma membrane in the form of nanovesicles. We propose that this process is the result of the self-adaptation of the membrane surface area to the elastic stress imposed by the spectrin cytoskeleton, via the local buckling of membrane under increasing cytoskeleton stiffness. This model introduces the concept of force balance as a regulatory process at the cell membrane and quantitatively reproduces the rate of area loss in aging red-blood cells.     

Electromagnetic Forces on Plasma Particles in the Presence of Resonant and Nonresonant Plasma Turbulence

Nonlinear forces on plasma particles in the presence of a test nonresonant wave and resonant plasma wave turbulence are calculated. The important feature of the considered nonlinear effect is that the forces due to the nonresonant test wave act on the plasma particles in the absence of linear and nonlinear resonances between the wave and the particles. Although in a closed plasma-wave system the process is balanced by the quasilinear interaction between the plasma resonant turbulence and plasma particles (leading to nonstationarity and inhomogeneity of the system), in open systems the effect can be significant.     

高密度等离子体工艺总体模型初探

介绍了与高密度等离子体工艺相关的模型和数值模拟方法,即连续流和动力学方法。在漂流－扩散方程的连续流模型和单元粒子／蒙特卡罗碰撞动力学模型的基础上,提出了一个等离子体工艺模型。讨论了对等离子体鞘层、等离子体刻蚀和淀积过程的模拟方法,提出了一个高密度等离子体工艺总体模型的初步方案。     

Effect of interparticle forces on the coagulation of ultrafine SiO2 particles

The mechanism of ultrafine (<100 nm) SiO₂ particle production under thermal arc plasma conditions is studied by modelling. Two cases of the process are considered in the model: (i) when it is determined as a pure free-molecular coagulation; (ii) when the coagulation is influenced by the interparticle forces. The Hamaker formula is used to present the van der Waals forces between the particles. Particle size distribution functions (PSDF) are calculated for both cases. It is shown that inclusion of the interparticle interactions does not affect the self-preservation of the PSDF. The mean particle sizes are obtained from the PSDF and compared. Higher values are observed in the case that includes van der Waals forces. Comparison between experimental and calculated PSDF shows better agreement in the case considering interparticle forces.     

On the dissipation of mechanical energy in a noncontact dynamic mode of a scanning probe microscope under vacuum

This paper presents a critical review of experiments on the measurement of conservative and dissipative forces in dynamic contacts of nanotips of scanning probe microscopes with surfaces (under high-vacuum conditions). In particular, it is noted that, although the perpendicular and parallel oscillatory motions of tips in the vicinity of the surface of various materials have been investigated experimentally, the question as to the nature of noncontact damping forces at distances ≥0.5 nm remains open. A phenomenological relationship between arbitrary conservative and dissipative forces acting on the tip is proposed. This relationship includes a model parameter characterizing the rate or time of the dissipative process and offers a correct explanation of the observed damping. It is shown that, despite the large differences in the damping coefficients, geometric sizes of tips, and types of conservative interactions in different experiments, the dissipative forces in perpendicular and parallel oscillatory motions of tips are adequately described within the proposed theory.     

Using response surface methodology for optimizing deposited partially stabilized zirconia in plasma spraying

This paper presents the application of the surface response methodology and fractional factorial experiment in developing a robust partially stabilized zirconia (PSZ) coating in plasma spraying processes. In this study, a two-step experimental design with a nonlinear regression model is proposed. After conducting a series of initial experiments in a controlled environment, significant factors for plasma spraying processes are selected to construct an appropriate response surface model for developing a robust performance for plasma spraying processes.The experimental results show that using a quadric response surface model with the proposed two-step design strategy is a simple, effective and efficient approach. In the optimized model, several response plots were generated to examine parameter effects on the profile hardness. Optimization of the PSZ performances in a plasma spray process has been performed.     

Experimental Investigation of Jet Stabilization in Near-Axis Region of Plasma Reactor

The paper presents the results of experimental investigations of aerodynamics and heat transfer in a vortex plasma reactor with gas jet injection along its axis. The process of mixing under quasi-isothermal conditions (injection of slightly heated jet) and inhomogeneous conditions (injection of helium and argon) as well as with the injection along the axis of plasma jet is studied consistently. A detailed comparison of temperature fields and concentrations is presented for these conditions. The process of jet expansion is shown to be defined mainly by three factors — centrifugal mass forces, buoyancy forces and initial velocity shift in a jet at the chamber inlet. The end flows contribute considerably to the process of jet mass transfer with peripheral flow.     

Limiting role of desorption in hydrogen transport across a deposited beryllium film

Hydrogen transport across a deposited beryllium layer has been investigated using the hydrogen permeability and concentration pulse methods. A layer of beryllium was deposited on a prepurified nickel membrane by cathode sputtering in a glow discharge plasma in “especially pure” grade hydrogen. An analysis of the experimental results showed that the main limiting process for hydrogen transport is desorption from the layer rather than diffusion in the bulk of the layer. A mathematical transport model is proposed and used to determine the rate constant of hydrogen desorption from beryllium. Zh. Tekh. Fiz. 68, 128–130 (January 1998)