细胞离子在振荡电磁场作用下的受力模型分析

本文通过生物细胞模型,研究振荡电场、振荡磁场以及振荡磁场产生的感应电场对细胞离子的作用机理。模型分析结果表明,电场力和罗仑兹力对细胞膜两侧的自由离子将产生加速度,振荡离子将产生周期性电位移。该模型同时也解释了脉冲电磁场比同参教的连续场产生更多的生物效应,以及连续场在开始施加和切除时的效应最大。     

Mechanism for action of electromagnetic fields on cells

A biophysical model for the action of oscillating electric fields on cells, presented by us before [Biochem. Biophys. Res. Commun. 272(3) (2000) 634-640], is extended now to include oscillating magnetic fields as well, extended to include the most active biological conditions, and also to explain why pulsed electromagnetic fields can be more active biologically than continuous ones. According to the present theory, the low frequency fields are the most bioactive ones. The basic mechanism is the forced-vibration of all the free ions on the surface of a cell's plasma membrane, caused by an external oscillating field. We have shown that this coherent vibration of electric charge is able to irregularly gate electrosensitive channels on the plasma membrane and thus cause disruption of the cell's electrochemical balance and function [Biochem. Biophys. Res. Commun. 272(3) (2000) 634-640]. It seems that this simple idea can be easily extended now and looks very likely to be able to give a realistic basis for the explanation of a wide range of electromagnetic field bioeffects.     

Protein translocation across membranes.

Cellular membranes act as semipermeable barriers to ions and macromolecules. Specialized mechanisms of transport of proteins across membranes have been developed during evolution. There are common mechanistic themes among protein translocation systems in bacteria and in eukaryotic cells. Here we review current understanding of mechanisms of protein transport across the bacterial plasma membrane as well as across several organelle membranes of yeast and mammalian cells. We consider a variety of organelles including the endoplasmic reticulum, outer and inner membranes of mitochondria, outer, inner, and thylakoid membranes of chloroplasts, peroxisomes, and lysosomes. Several common principles are evident: (a) multiple pathways of protein translocation across membranes exist, (b) molecular chaperones are required in the cytosol, inside the organelle, and often within the organelle membrane, (c) ATP and/or GTP hydrolysis is required, (d) a proton-motive force across the membrane is often required, and (e) protein translocation occurs through gated, aqueous channels. There are exceptions to each of these common principles indicating that our knowledge of how proteins translocate across membranes is not yet complete.     

Mechanical state of airway smooth muscle at very short lengths.

Although the shortening of smooth muscle at physiological lengths is dominated by an interaction between external forces (loads) and internal forces, at very short lengths, internal forces appear to dominate the mechanical behavior of the active tissue. We tested the hypothesis that, under conditions of extreme shortening and low external force, the mechanical behavior of isolated canine tracheal smooth muscle tissue can be understood as a structure in which the force borne and exerted by the cross bridge and myofilament array is opposed by radially disposed connective tissue in the presence of an incompressible fluid matrix (cellular and extracellular). Strips of electrically stimulated tracheal muscle were allowed to shorten maximally under very low afterload, and large longitudinal sinusoidal vibrations (34 Hz, 1 s in duration, and up to 50% of the muscle length before vibration) were applied to highly shortened (active) tissue strips to produce reversible cross-bridge detachment. During the vibration, peak muscle force fell exponentially with successive forced elongations. After the episode, the muscle either extended itself or exerted a force against the tension transducer, depending on external conditions. The magnitude of this effect was proportional to the prior muscle stiffness and the amplitude of the vibration, indicating a recoil of strained connective tissue elements no longer opposed by cross-bridge forces. This behavior suggests that mechanical behavior at short lengths is dominated by tissue forces within a tensegrity-like structure made up of connective tissue, other extracellular matrix components, and active contractile elements.     

Ion channels activated by specific Ti or T3 antibodies in plasma membranes of human T cells.

T lymphocytes are activated to proliferate via a surface membrane receptor recognizing the antigen/major histocompatibility complex. This membrane component is comprised of at least five polypeptide subunits, collectively termed the Ti-T3 receptor complex. A transient increase in cytosolic free calcium occurs as an early event in the T-cell activation process and is necessary for induction of the endogenous IL-2 and certain other genes. Monoclonal antibodies specific to epitopes of either the Ti or the T3 components were shown to be effective agonists, also leading to such transient rises in cytosolic free calcium and activating the lymphocytes. Here we show, using micropipette-supported bilayers formed from membranes of the human T-cell line REX, that Ti- or T3-specific antibodies cause opening of ligand gated ion channels. Both types of specific antibodies yielded similar histograms of conductance amplitudes which show a channel with a conductance of 2-3 pS in symmetrical 100 mM CaCl2 solutions. These channels allow the passage of calcium and barium ions and are blocked by lanthanum ions, suggesting that they are specific for calcium. We propose that these channels, by allowing the entry of external calcium, may account for a large fraction of the rise in intracellular calcium observed upon triggering of the Ti-T3 receptor.     

Fabrication and optimal design of differential electromagnetic transducer for implantable middle ear hearing device

A differential electromagnetic transducer (DET), with similar frequency characteristics to those of a normal middle ear, is designed and implemented for use in an implantable middle ear hearing device (IMEHD). To optimize the characteristics of the DET that depend on the electromagnetically forced vibration, a theoretical analysis is conducted to design the vibrating part. The electromagnetic force of the DET is simulated according to the design parameters of the coil size using a finite element analysis (FEA). As a result, the maximal vibration force is achieved when the optimal length and thickness of the cylindrical coil is 70% of the length of the magnets and 56% of their radius. The vibration characteristics of the DET are then simulated when applying the maximal force. The optimally designed DET is implemented using MEMS technology and vibration experiments carried out with the fabricated DET in an unloaded state. The vibrating displacement of the DET is about 200 nm within a range between 0.1 and 1.5 kHz when a current of 1 mA(rms) is applied to the coil. To investigate the usefulness of the DET, in vitro and in vivo experiments are conducted using the ossicular chain of a cadaver and guinea pig, and the results verify that the implemented DET performs well as a transducer for an IMEHD.     

A Mathematical Model of the Human Circadian System and Its Application to Jet Lag

A mathematical model of the circadian system is described that is appropriate for application to jet lag. The core of the model is a van der Pol equation with an external force. Approximate solutions of this equation in which the external force is composed of a constant and an oscillating term are investigated. They lead to analytical expressions for the amplitude and period of free-running rhythms and for the frequency limits of the entrainment region. The free-running period increases quadratically with stiffness. Both period and amplitude depend on the value of the constant external force. The width of the range of entrainment is mostly determined by the external force, whereas the relative position of this range follows the intrinsic period of the oscillator. Experiments with forced and spontaneous internal desynchronization were evaluated using these analytical expressions, and estimates were obtained for the intrinsic period of the oscillator, its stiffness, and the external force. A knowledge of these model parameters is essential for predictions about circadian dynamics, and there are practical implications for the assessment of the adaptation after rapid transmeridian travel.     

A Mathematical Model of the Human Circadian System and Its Application to Jet Lag

A mathematical model of the circadian system is described that is appropriate for application to jet lag. The core of the model is a van der Pol equation with an external force. Approximate solutions of this equation in which the external force is composed of a constant and an oscillating term are investigated. They lead to analytical expressions for the amplitude and period of free-running rhythms and for the frequency limits of the entrainment region. The free-running period increases quadratically with stiffness. Both period and amplitude depend on the value of the constant external force. The width of the range of entrainment is mostly determined by the external force, whereas the relative position of this range follows the intrinsic period of the oscillator. Experiments with forced and spontaneous internal desynchronization were evaluated using these analytical expressions, and estimates were obtained for the intrinsic period of the oscillator, its stiffness, and the external force. A knowledge of these model parameters is essential for predictions about circadian dynamics, and there are practical implications for the assessment of the adaptation after rapid transmeridian travel.     

Magnitude of the protonmotive force in respiring Staphylococcus aureus and Escherichia coli.

The membrane potential and pH gradient developed across the plasma membranes of whole cells of Staphylococcus aureus and spheroplasts of Escherichia coli were estimated. The distributions of potassium ions in the presence of valinomycin and the pH gradient across the membrane were determined from the changes in pK and pH observed in the external medium during transition from the energized respiring state to the de-engerized resting condition. The protonmotive force in respiring cells was estimated at 211 mV for S. aureus and 230 mV for E. coli at external pH values of approximately 6.5. The adequacy of these protonmotive forces as a driving force for substrate accumulation or adenosine 5'-triphosphate synthesis is discussed.     

Development of an Exactly Solvable Model of Resonance Tunneling of Electromagnetic Waves through Gradient Barriers in a Nonuniform Magnetoactive Plasma

An exactly solvable one-dimensional model describing resonance tunneling (reflectionless transmission) of a transverse electromagnetic wave through wide layers of magnetoactive plasma is developed on the basis of the Helmholtz equation. The plasma layers include a set of spatially localized density structures the amplitudes and thicknesses of which are such that approximate methods are inapplicable for their analysis. The profiles of the plasma density structures strongly depend on the choice of the free parameters of the problem that determine the amplitudes of plasma density modulation, characteristic scale lengths of the density structures, their number, and the total thickness of the nonuniform plasma layer. The plasma layers can also include a set of random inhomogeneities. The propagation of electromagnetic waves through such complicated plasma inhomogeneities is analyzed numerically within the proposed exactly solvable model. According to calculations, there are a wide set of inhomogeneous structures for which an electromagnetic wave incident from vacuum can propagate through the plasma layer without reflection, i.e., the complete tunneling of thick plasma barriers takes place. The model also allows one to exactly solve a one-dimensional problem on the nonlinear transillumination of a nonuniform plasma layer in the presence of cubic nonlinearity. It is important that, due to nonlinearity, the thicknesses of the evanescent plasma regions can decrease substantially and, at a sufficiently strong nonlinearity, such regions will disappear completely. The problem of resonance tunneling of electromagnetic radiation through gradient wave barriers is of interest for various applications, such as efficient heating of dense plasma by electromagnetic radiation and transmission of electromagnetic signals from a source located in the near-Earth plasma or deep in the plasma of an astrophysical object through the surrounding evanescent regions.     

Optical measurements of Na-Ca-K exchange currents in intact outer segments isolated from bovine retinal rods

The properties of Na-Ca-K exchange current through the plasma membrane of intact rod outer segments (ROS) isolated from bovine retinas were studied with the optical probe neutral red. Small cellular organelles such as bovine ROS do not offer an adequate collecting area to measure Na-Ca-K exchange currents with electrophysiological techniques. This study demonstrates that Na-Ca-K exchange current in bovine ROS can be measured with the dye neutral red and dual-wavelength spectrophotometry. The binding of neutral red is sensitive to transport of cations across the plasma membrane of ROS by the effect of the translocated cations on the surface potential of the intracellular disk membranes (1985. J. Membr. Biol. 88: 249-262). Electrogenic Na+ fluxes through the ROS plasma membrane were measured with a resolution of 10(5) Na+ ions/ROS per s, equivalent to a current of approximately 0.01 pA; maximal electrogenic Na-Ca-K exchange flux in bovine ROS was equivalent to a maximal exchange current of 1-2 pA. Electrogenic Na+ fluxes were identified as Na-Ca-K exchange current based on a comparison between electrogenic Na+ flux and Na(+)-stimulated Ca2+ release with respect to flux rate, Na+ dependence, and ion selectivity. Neutral red monitored the net entry of a single positive charge carried by Na+ for each Ca2+ ion released (i.e., monitored the Na-Ca-K exchange current). Na-Ca-K exchange in the plasma membrane of bovine ROS had the following properties: (a) Inward Na-Ca-K exchange current required internal Ca2+ (half-maximal stimulation at a free Ca2+ concentration of 0.9 microM), whereas outward Na-Ca-K exchange current required both external Ca2+ (half-maximal stimulation at a free Ca2+ concentration of 1.1 microM) and external K+. (b) Inward Na-Ca-K exchange current depended in a sigmoidal manner on the external Na+ concentration, identical to Na(+)-stimulated Ca2+ release measured with Ca(2+)-indicating dyes. (c) The neutral red method was modified to measure Ca(2+)-activated K+ fluxes (half-maximal stimulation at 2.7 microM free Ca2+) via the Na-Ca-K exchanger in support of the notion that the rod Na-Ca exchanger is in effect a Na-Ca-K exchanger. (d) Competitive interactions between Ca2+ and Na+ ions on the exchanger protein are described.     

Linker-gating ring complex as passive spring and Ca(2+)-dependent machine for a voltage- and Ca(2+)-activated potassium channel

Ion channels are proteins that control the flux of ions across cell membranes by opening and closing (gating) their pores. It has been proposed that channels gated by internal agonists have an intracellular gating ring that extracts free energy from agonist binding to open the gates using linkers that directly connect the gating ring to the gates. Here we find for a voltage- and Ca(2+)-activated K+ (BK) channel that shortening the linkers increases channel activity and lengthening the linkers decreases channel activity, both in the presence and absence of intracellular Ca2+. These observations are consistent with a mechanical model in which the linker-gating ring complex forms a passive spring that applies force to the gates in the absence of Ca2+ to modulate the voltage-dependent gating. Adding Ca2+ then changes the force to further activate the channel. Both the passive and Ca(2+)-induced forces contribute to the gating of the channel.     

Configuration analysis and optimization on multipolar Galatea trap

Multipolar Galatea magnetic trap simulation model was established with the finite element simulation software COMSOL Multiphysics. Analyses about the magnetic section configuration show that better magnetic configuration should make more plasma stay in the weak magnetic field rather than the annular magnetic shell field. Then an optimization model was established with axial electromagnetic force, weak magnetic field area and average magnetic mirror ratio as the optimization goals and with the currents of myxines as design variables. Select appropriate weight coefficients and get optimization results by applying genetic algorithm. Results show that the superiority of the target value of typical application parameters, including the average magnetic mirror can reduce more than 5%, the weak magnetic field area can increase at least 65%, at the same time, axial electromagnetic force acting on the outer myxines can be reduced to less than 50 N. Finally, the results were proved by COMSOL Multiphysics and the results proved the optimized magnetic trap configuration with more plasma in the weak magnetic field can reduce the plasma diffusion velocity and is more conducive for the constraint of plasma.     

Mechanical vibration model for chromosomes in metaphase of mitosis and possible application to the interruption of cell division

The behavior of chromosomes subjected to external mechanical vibration in metaphase of mitosis is modeled, and its possible applications to the interruption of cell divisions is investigated. The modeling is based on a mass and string coupled system where the mass and the string correspond to the condensed chromosome and the kinetochore microtubule, respectively. After establishing the motion of equation for a chromosome, the frequency response to the forced vibration from the outside of the cell is formulated. This will lead to an efficient method for kinetic energy transfer to chromosomes by finding the resonant frequencies of the cell system. There is a possibility that the increased kinetic energy in one chromosome will affect the cell as a whole, thereby interrupting the mitosis by the mechanisms that are discussed in this report. Rough calculation shows that the vibration in the frequency range of 22-50 kHz is effective for controlling the mitosis of human cells.     

The sarcoplasmic calcium pump — A most efficient ion translocating system

In contrast to the sodium-potassium transporting plasma membranes, the sarcoplasmic membranes (SR) are highly specialized structures into which only two major intrinsic proteins, a calcium transporting protein and a calcium binding protein are embedded. The calcium transporting protein is a highly asymmetric molecule. It binds two calcium ions with a very high affinity at its external, and two calcium ions with low affinity at the internal section of the molecule. ATP is bound with high affinity to an external binding site, inducing a conformational change. When the vesicular membranes are exposed to solutions containing Ca⁺⁺, Mg⁺⁺ and ATP, ATP is hydrolyzed and simultaneously calcium ions are translocated from the external medium into the vesicular space. When calcium ions are translocated in the opposite direction, ATP is synthesized. The calcium-ATP ratio for ATP cleavage as well as for ATP synthesis is 2. Thus, the SR membranes can transform reversibly chemical into osmotical energy. Inward and outward movements of calcium ions are relatively slow processes connected with the appearance and disappearance of different phosphorylated intermediates. One phosphorylated intermediate is formed by phosphoryltransfer from ATP when calcium ions are present in the medium. In contrast, when calcium ions are absent from the external medium, two different intermediates can be formed by the incorporation of inorganic phosphate. Only when calcium ions present in the internal space of the vesicles are released, the incorporation of inorganic phosphate gives rise to an intermediate whose phosphoryl group can be transferred to ADP.Presented at the EMBO-Workshop on Transduction Mechanism of Photoreceptors, Jülich, Germany, October 4–8, 1976     

Effect of Tendon Vibration on Hemiparetic Arm Stability in Unstable Workspaces

Sensory stimulation of wrist musculature can enhance stability in the proximal arm and may be a useful therapy aimed at improving arm control post-stroke. Specifically, our prior research indicates tendon vibration can enhance stability during point-to-point arm movements and in tracking tasks. The goal of the present study was to investigate the influence of forearm tendon vibration on endpoint stability, measured at the hand, immediately following forward arm movements in an unstable environment. Both proximal and distal workspaces were tested. Ten hemiparetic stroke subjects and 5 healthy controls made forward arm movements while grasping the handle of a two-joint robotic arm. At the end of each movement, the robot applied destabilizing forces. During some trials, 70 Hz vibration was applied to the forearm flexor muscle tendons. 70 Hz was used as the stimulus frequency as it lies within the range of optimal frequencies that activate the muscle spindles at the highest response rate. Endpoint position, velocity, muscle activity and grip force data were compared before, during and after vibration. Stability at the endpoint was quantified as the magnitude of oscillation about the target position, calculated from the power of the tangential velocity data. Prior to vibration, subjects produced unstable, oscillating hand movements about the target location due to the applied force field. Stability increased during vibration, as evidenced by decreased oscillation in hand tangential velocity.     

Trapping single ions inside single ion channels.

Single Ca++-activated K+ channels from rat muscle plasma membranes are inhibited by Ba++. A single Ba++ entering the channel's conduction pore induces a long-lived blocked state. This study employs Ba++ as a probe of the channel's conduction pathway to show that the channel can be forced to close with a single Ba++ ion inside the pore. A Ba++ ion inside the closed channel is trapped and cannot escape until the channel opens. The results demonstrate that in the channel's closed state, the cytoplasmic side of the conduction pore is obstructed to the passage of ions.     

Role of membranes in disease

The membranes of mammalian cells are composed of an ordered array of lipids and proteins, the latter containing carbohydrate residues directed towards the exterior and important in the interaction of cells with each other and with external proteins. This external (plasma) membrane and other more simple membranes within the cell are damaged in all diseases which compromise the integrity of the cell. However, in many cases, chemical or functional changes in these membranes are central to the pathogenesis of the disease. These processes are illustrated, and a classification of membrane-related diseases is proposed. This includes: receptor-related diseases such as type II familial hypercholesterolaemia, Grave's disease, some lysosomal storage diseases and some forms of diabetes and obesity; structural instability as manifested by red cell abnormalities and multiple sclerosis; changes in lipid state as in muscular dystrophy and multiple sclerosis; altered permeability or transport as in cystic fibrosis, diseases associated with specific transport defects, and the action of many bacterial toxins, and abnormality of the cytoskeleton-membrane interface as in Chediak-Higashi disease and some diseases associated with red cell abnormalities. Different mechanisms can contribute to the membrane disorder in a single disease state and many of these are described to illustrate this diversity.     

The origin of membranes

The physicochemical conditions of the environment in which life arose are discussed, along with the appearance of protocells, their membranous envelope and the subsequent appearance of plasma membranes. The hypothesis that the first cells originated in reservoirs where potassium and magnesium salts (necessary for protein synthesis and thus for the formation of a cellular membrane) dominated, is substantiated. This was followed by adaptation of these cells to an external ocean-like environment, where sodium salts were prevalent. This stage of evolution required a plasma membrane capable of providing ion asymmetry between the cell’s cytoplasm and the external environment. At this stage of evolution in the predecessors of animals, the process of removal of sodium ions and accumulation of potassium ions began functioning in the plasma membrane. The problem of multicellular organisms was solved differently by animals and plants: animals developed a system of the extracellular fluids that provided stable physicochemical conditions on the external surface of the plasma membrane. Sodium ions were the stimulus for the formation of the polar cell, where sodium channels are situated on one side of the plasma membrane, and sodium pumps on the other, allowing the development of the absorption, excretion and breathing functions. The formation of fluids of the internal environment enabled the development of homeostasis and facilitated the biological progress of the animal kingdom.     

Structurally distinct plasma membrane regions give rise to extracellular membrane vesicles in normal and transformed lymphocytes

Shedding of extracellular membranes from the cell surface may be one of the means through which cells communicate with one another. In an attempt to elucidate whether cell surface exfoliation is a directed or random process, we investigated the membrane lipid and protein composition and membrane lipid order of shed extracellular membranes and of plasma membranes from which they arose in normal circulating lymphocytes and in the B-lymphoblastoid cell lines Raji, WI HF2 729 and the T-lymphoblastoid cell line Jurkat. Extracellular membranes derived from transformed cell lines were more rigid as assessed by steady state polarization of 1,6-diphenylhexatriene (DPH) and were highly enriched in cholesterol when compared with the corresponding plasma membrane. The extracellular membranes from normal lymphocytes, on the other hand, were more fluid and contained more polyunsaturated acyl chains than did the plasma membranes from these cells. Our results suggest that extracellular membranes are shed from specialized regions of the lymphocyte plasma membrane and that membrane exfoliation is likely to be a directed event.