The effect of pressure on the electrical breakdown in the membranes of Valonia utricularis

The interpretation of electrical breakdown in terms of electro-mechanical instabilities, predicts that the breakdown potential should decrease with increasing cell turgor pressure. Experiments were conducted to test this hypothesis on cells of Valonia utricularis over a turgor pressure range of 0.5-10(5)-5.0-10(5) N/m2. Electrical breakdown was measured using intracellular electrodes and 500 mus current pulses. The pressure was monitored by an intracellular micropipette pressure transducer. The results obtained show a linear decrease in the critical breakdown potential with pressure. The effective compressive modulus of the cell membrane, gamma, is calculated from the slope of this line to 69+/-10-10(5) N/m2 (average value of seven measurements). This is consistent with the theoretical prediction of the electromechanical model using our previously determined values of the elastic modulus of the membrane. A theoretical analysis is given of the effects of pressure on the breakdown, This includes also considerations of the indirect effect of pressure on the membrane via stretching of the cell wall with a possible coupling of such strains to the cell membrane. The results and analysis presented allow us to conclude on the basis of the experimentally determined breakdown P.K. of 959 mV that the region of membrane where electrical breakdown occurs is a dielectric with one of the following combinations of parameters: (A) a thickness delta=7-9 nm with a dielectric constant epsilon=greater than 10, e.g. a hydrated protein spanning the whole membrane. (B) delta=4-5 nm with epsilon=3-8, e.g. a lipoprotein of lipid bilayer dimensions. (C) delta approximately 2 nm with epsilon=2-3, e.g. a half lipid bilayer. If we assume that the breakdown P.D. of the tonoplast and plasmalemma are identical, that is 480 mV, then there is only one reasonable choice for the membrane thickness and the dielectric constant: delta=2 nm, epsilon=3-8, e.g. a (lipo) proteinaceous module facing a half life lipid bilayer.     

镀W碳纳米管增强Mg基复合材料的力学和电学性能

以羰基钨为前驱体,采用金属有机化学气相沉积在碳纳米管表面镀覆了金属W。利用磁力搅拌混粉和放电等离子体烧结制备了镀W碳纳米管(W-CNTs)/Mg复合材料,并研究了W-CNTs含量对材料力学性能和电导率的影响。结果表明:W-CNTs含量为0.75%(质量分数,下同)时,复合材料抗拉强度和维氏硬度最大,较纯Mg增加了40.1%和35.1%;基体中添加W-CNTs虽然使复合材料电导率下降,但0.75%W-CNTs/Mg复合材料电导率仍可达到纯Mg的94.8%;W-CNTs/Mg复合材料的力学性能和电导率均高于CNTs/Mg复合材料。     

Sphingomyelin breakdown and cell fate

A growing number of cell-surface receptors are now being shown to generate signals that trigger the hydrolysis of sphingomyelin to release diffusible ceramides. Ceramides have been implicated as key mediators in signaling pathways, with outcomes as diverse as cell proliferation, differentiation, growth arrest and apoptosis. The response depends on cell type, whether the signal is integrated with other signals originating from the same receptor and on the subcellular location of sphingomyelin hydrolysis and ceramide release.     

Freeze-fracturing studies on human erythrocyte membranes effected by external pulsed electrical field

Electron microscopic observations made on the freeze-fracturing replica of plasma membranes of human erythrocytes exposed to external pulsed electrical field have shown that under experimental conditions, some particles and fibers appear around the cells pulsed by higher intensities than 3 KV/cm. Electrophoretical analysis for the cell suspensions has proved that some of these particles and fibers are membrane proteins and membrane skeleton proteins escaped from cells. On account of this, the stability of cell membrane decreases and the cells are easy to change their shapes and to form pseudopodium like structures and protein free membrane lipids tend to form vesicular structure in cytoplasma. The changes of protein-protein interactions and protein-lipid interactions caused by electrical field are considered as the dominating mechanism of cell membrane electroporation. An argument about intramembrane particles (IMPS) and other possibilities of the contributions to IMPS, for instance, ice crystals formation which related to membrane hydrophilic or hydrophobic properties were discussed in this paper.     

矿山机电设备检修与优化措施

矿产资源的开发与矿山设备密切相关,广泛采用科学有效的矿山开采设备可以极大地提高矿山的开采效率,所以矿山开采所用的基础设备检修十分重要。本文从矿山设备故障判断和处理技术的现状出发,分析了矿山机电设备故障诊断的技术,着重论述了智能化诊断、数字模型诊断、仪器诊断和主观经验诊断四种技术,并针对这几种诊断方式的现状和应用,提出了一些建议和参考,期望能对矿山机电设备的检修和优化提供一些帮助。     

Optical and electrical studies on dansyllysine-valinomycin in thin lipid membranes

Dansyllysine-valinomycin, a fluorescent analogue of the ionophore valinomycin was synthesized and incorporated into black lipid membranes. Its concentration inside the membrane was measured fluorometrically and was also determined from electrical relaxation experiments, which were analyzed on the basis of a previously proposed carrier model. The results of both methods agreed within less than one order of magnitude. This appears satisfactory in view of the sources of error inherent in both procedures. A conductance increment per carrier molecule of about 3 - 10(-17) omega-1 was obtained for dansyllysine-valinomycin in diphytanoyllecithin membranes at 25 degrees C and 1 M RbCl in the aqueous phases. This is about 400 times smaller compared to unmodified valinomycin in monoolein membranes. The difference is mainly caused by the change in the membrane properties and to a smaller extent by the structural modification of the carrier.     

TGF-alpha-induced breakdown of stress fibers and degradation of tropomyosin in NRK cells is blocked by a proteasome inhibitor

Treatment of NRK cells with TGF-alpha in the presence of serum initiates disassembly of cytoskeletal stress fibers and suppresses the synthesis of tropomyosin isoforms (TMs) 1, 2, and 3 but not TMs 4 and 5 (Cooper et al., Cancer Res. 47, 4493-4500, 1987). In order to determine how the loss of tropomyosin is induced and what role it plays in cytoskeletal disruption, the turnover of tropomyosin was studied in the presence of the transforming growth factor and protease inhibitors. Cells were pulse-labeled with [35S]methionine and chased in the absence or the presence of the growth factor. It was found that TMs 1, 2, and 3 are degraded at about twice the rate of TMs 4 and 5 in control cells and that the rate of degradation of TMs 1-3 is accelerated by the growth factors. Degradation of TMs in control and growth factor-treated cells is blocked by a membrane-permeable inhibitor of cysteine proteases (LLnL) that acts upon calpains and proteasomes, and the cells maintain a flattened shape with a normal complement of stress fibers. Application of inhibitors that block calpains but not proteasomes does not block TM degradation. Treatments (suspension culture or cytochalasin B) that disrupt stress fibers without application of the growth factors also accelerate TM degradation, suggesting that acceleration of TM degradation is a consequence of its release from stress fibers during their breakdown. The normally more rapid turnover of the TM isoforms 1-3 that are lost in the phenotypically transformed cells could serve to facilitate the cytoskeletal reorganization that follows the activation of signal transduction pathways by the transforming growth factors observed in this study or during other rearrangements of the cytoskeleton such as occur during cell migration or mitosis.     

Contribution of mechanical activity and electrical activity to cellular electrical uncoupling in ischemic rabbit papillary muscle

We separated mechanical and electrical activity during ischemia and we assessed the contribution of each of these variables to ischemia induced cellular electrical uncoupling. In two groups of rabbit papillary muscles, mechanical and electrical activity at the onset of ischemia was abolished. We determined how this affected the time of onset of uncoupling. Also, action potential duration (APD80), conduction velocity and extracellular pH were recorded. In the first group, the BDM group, mechanical arrest was achieved with 10 mM BDM (2,3-butanedione monoxime). This had no effect on APD80 and conduction velocity. In the second group, the quiescent group, electrical as well as mechanical arrest was obtained by not stimulating the muscles. The effect of electrical arrest was estimated as the difference between this group and the BDM group. Mechanical arrest delayed uncoupling significantly (control: 14.6 +/- 0.9 (min), mean +/- S.E.M., n = 10; BDM: 19.3 +/- 1.1, n = 10; P < 0.025). Electrical arrest caused an even greater delay (quiescent: 41.6 +/- 4.6, n = 7; P < 0.001 v control and BDM). In the BDM group, these effects were not associated with a difference of APD80 during ischemia, but with a smaller decrease of conduction velocity. Delay of uncoupling correlated with delay of extracellular acidosis. We conclude: (1) mechanical and electrical activity both are significant determinants of ischemia induced uncoupling; (2) delay of uncoupling is associated with delay of extracellular acidosis, indicating a reduced rate of metabolism.     

Phosphatidylinositol 3-kinase is necessary and sufficient for insulin-stimulated stress fiber breakdown

Rat-1 fibroblasts overexpressing the human insulin receptor undergo rapid actin rearrangement in response to insulin. Breakdown of stress fibers present in quiescent cells is followed by transient membrane ruffling and a return of stress fibers. We investigated the signaling pathways that mediate this insulin-stimulated reorganization of the actin cytoskeleton, which was visualized with rhodamine-phalloidin. Treatment of cells with the phosphatidylinositol 3-kinase (PI3-kinase) inhibitor wortmannin prevented insulin action at the preliminary step of stress fiber breakdown. Cellular microinjection of a polyclonal antibody directed against the p85 subunit of PI3-kinase as well as a purified recombinant p85-SH2 domain protein also inhibited actin reorganization. Transient expression of a constitutively active form of PI3-kinase (p110*) was sufficient to cause both stress fiber breakdown and membrane ruffling in the absence of insulin. Microinjection of a polyclonal anti-Shc antibody or dominant negative N17-Ras protein did not affect actin dynamics, and although constitutively active V12-Ras caused modest cytoskeletal reorganization, this effect was blocked by pretreatment with wortmannin. In summary, activation of PI3-kinase is necessary and sufficient to stimulate actin rearrangement, indicating that PI3-kinase may initiate the only signaling cascade required for insulin to induce cytoskeletal restructuring.     

Vibrational stimulation of the skin using both electrical and mechanical stimuli

Pulmonary eosinophilic granuloma diagnosed by lung biopsy is described; the case demonstrates the remission and relapse that may occur.     

The effect of oxidative stress on structural transitions of human erythrocyte ghost membranes

The influenza virus hemagglutinin (HA) contains three highly conserved cysteine residues at positions 551, 559, and 562 close to the carboxyl-terminus of the HA2 subunit which serve as palmitylation sites. Wild-type HA of influenza virus A/FPV/Rostock/34 (H7N1) and HA permutated by exchange of the acylated cysteine to serine residues were expressed in CV-1 cells by a SV40 vector system. Since density of immunostained HA on the cell surface measured by flow cytometric analysis did not differ between wild-type and acylation mutants, it was possible to compare acylation mutants and wild-type HA for their capacity to induce membrane fusion at low pH. The following observations were made: (1) lateral diffusion of a lipid-like fluorophore (R-18) from the erythrocyte membrane to the plasma membrane of cells expressing HA on the surface occurred equally well with mutants and wild type. (2) Diffusion of a low-molecular-weight fluorescent water-soluble probe (calcein) from erythrocytes into the cytoplasm of HA-expressing cells was not altered either. (3) However, depending on the position and the number of the deleted acylation sites, the mutants showed a reduced ability to induce syncytia. The data indicate that deacylation of the cytoplasmic tail has no measurable effect on the capacity of HA to induce membrane fusion and pore formation but that it suppresses syncytia formation.     

Myosin synthesis increased by electrical stimulation of skeletal muscle cell cultures

When cultures of skeletal muscle cells of the chick embryo are subjected to repetitive, electrical stimulation, the contractions increase the amount of protein produced by these cells. The increase is greater for contractile proteins such as myosin heavy chain than for total cellular protein. This demonstrates that in a culture system of skeletal muscle cells that have differentiated in the absence of innervation, one can elicit the protein synthetic response associated with skeletal muscle hypertrophy in vivo.