Anaesthetic modulation of nicotinic ion channel kinetics in bovine chromaffin cells

1. We have investigated the action of the anaesthetics methoxyflurane, methohexitone and etomidate on the nicotinic acetylcholine receptor channel of bovine adrenal chromaffin cells using the whole cell patch clamp technique. 2. Spectral analysis of macroscopic currents evoked by 25 microM carbachol revealed that each of the agents tested reduced the lifetime of the channel open state in a dose-dependent manner. The whole cell current was inhibited in a concentration-dependent fashion by each agent. 3. Channel gating parameters were calculated from single channel studies and the results used to test models explaining the modulation of nicotinic acetylcholine receptor channels by anaesthetics. 4. Each of the agents studied reduced the mean channel open time in a concentration-dependent manner. Anaesthetic concentrations reducing mean open time by 50% were: 370 microM methoxyflurane, 30 microM methohexitone or 23 microM etomidate. 5. Methohexitone and etomidate produced an increase in the number of brief closures within bursts, while no such increase was observed with methoxyflurane. Despite these inter-burst gaps, mean burst length was reduced by each of the agents tested. 6. It is concluded that a simple sequential blocking model fails to account for the action of these anaesthetics. An extended model, in which blocked channels can close, may be applicable.     

Gramicidin channels in phospholipid bilayers with unsaturated acyl chains

In organic solvents gramicidin A (gA) occurs as a mixture of slowly interconverting double-stranded dimers. Membrane-spanning gA channels, in contrast, are almost exclusively single-stranded beta(6,3)-helical dimers. Based on spectroscopic evidence, it has previously been concluded that the conformational preference of gA in phospholipid bilayers varies as a function of the degree of unsaturation of the acyl chains. Double-stranded pi pi(5,6)-helical dimers predominate (over single-stranded beta(6,3)-helical dimers) in lipid bilayer membranes with polyunsaturated acyl chains. We therefore examined the characteristics of channels formed by gA in 1-palmitoyl-2-oleoylphosphatidylcholine/n-decane, 1,2-dioleoylphosphatidylcholine/n-decane, and 1,2-dilinoleoylphosphatidylcholine/n-decane bilayers. We did not observe long-lived channels that could be conducting double-stranded pi pi(5,6)-helical dimers in any of these different membrane environments. We conclude that the single-stranded beta(6,3)-helical dimer is the only conducting species in these bilayers. Somewhat surprisingly, the average channel duration and channel-forming potency of gA are increased in dilinoleoylphosphatidylcholine/n-decane bilayers compared to 1-palmitoyl-2-oleoylphosphatidylcholine/n-decane and dioleoylphosphatidylcholine/n-decane bilayers. To test for specific interactions between the aromatic side chains of gA and the acyl chains of the bilayer, we examined the properties of channels formed by gramicidin analogues in which the four tryptophan residues were replaced with naphthylalanine (gN), tyrosine (gT), and phenylalanine (gM). The results show that all of these analogue channels experience the same relative stabilization when going from dioleoylphosphatidylcholine to dilinoleoylphosphatidylcholine bilayers.     

Gramicidin A aggregation in supported gel state phosphatidylcholine bilayers

Using an atomic force microscope, supported bilayers of saturated phosphatidylcholine (in the gel state) containing various amounts of gramicidin A (gA) were imaged in aqueous solutions and at room temperature. gA clusters were directly observed for the first time under these conditions. It was found that, at a lower gA concentration, gA aggregated into domains, composed of small clusters along with a considerable amount of lipids. This basic aggregation unit, most likely a hexamer, remained the same for acyl chain lengths from 14 to 18 carbons. These small clusters were observed to form elongated aggregates (line type) but never into extended pure gA domains. When gA concentrations were increased, for bilayers with 16 carbons or less, gA aggregated into larger domains but the basic unit remained separated by lipid molecules. At about 5 mol % gA, a percolation-like transition occurred at which the line type aggregates were connected to each other. However, for bilayers with more than 16 carbons, multiple lamellar structures were formed at higher gA fractions and the top layer had a ripple-like surface morphology. The molecular mechanism for the formation of these peculiar structures remains to be elucidated.     

The stresses in a face-centered cubic single crystal under uniaxial tension

Finite element method (FEM) analysis was employed to examined the effect of end constraints on elastic and elasto-plastic stress and strain nonuniformities which develop in Cu single crystals. When no end constraints are applied, stresses are uniformly distributed throughout the specimen. Under end constraints, nonuniformities develop which can extend over considerable portions of the specimen and are strongly dependent on the ratio of diameter to length and on orientation. Results indicate that stress and strain nonuniformities are likely to be most significant for fatigue testing of single crystals.     

单轴拉伸作用下岩石裂隙起裂扩展的数值模拟

地下采矿工程开挖对天然岩石是一种卸荷行为,卸荷后的岩石容易受到拉应力作用,造成岩石内部原生裂隙的扩张贯通,从而极大影响工程岩体稳定性。为研究拉伸作用下岩石内部裂隙的萌生演化规律,采用扩展有限单元法,对含不同裂隙倾角的岩石进行了模拟研究。结果表明:拉伸应力下岩石裂隙起裂主要受Ⅰ型应力强度因子控制,裂隙倾角越大,起裂越困难,相应的起裂角也越大;裂隙扩展时的扩展角保持不变,扩展方向与最大拉应力方向保持垂直。     

Microstructure evolution in Zr under equal channel angular pressing

Pure polycrystalline Zr was deformed by equal channel angular pressing (ECAP), and the microstructural characteristics were analyzed. By repeated alternating ECAP, it was possible to refine the grain size from 200 to 0.2 μm. Subsequent annealing heat treatment at 550 °C resulted in a grain growth of up to 6 μm. Mechanical twinning was an important deformation mechanism, particularly during the early stage of deformation. The most active twinning system was identified as 85.2 deg {10 2}〈 011〉 tensile twinning, followed by 57.1 deg {10 1}〈 012〉 compressive twinning. Crystal texture as well as grain-boundary misorientation distribution of deformed Zr were analyzed by X-ray diffraction (XRD) and electron backscattered diffraction (EBSD). The ECAP-deformed Zr showed a considerable difference in the crystallographic attributes from those of cold-rolled Zr or Ti, in that texture and boundary misorientation-angle distribution tend toward more even distribution with a slightly preferential distribution of boundaries of a 20 to 30 deg misorientation angle. Furthermore, unlike the case of cold rolling, the crystal texture was not greatly altered by subsequent annealing heat treatment. Overall, the present work suggests ECAP as a viable method to obtain significant grain refining in hexagonal close-packed (hcp) metals. This article is based on a presentation made in the symposium entitled “Defect Properties and Mechanical Behavior of HCP Metals and Alloys” at the TMS Annual Meeting, February 11–15, 2001, in New Orleans, Louisiana, under the auspices of the following ASM committees: Materials Science Critical Technology Sector, Structural Materials Division, Electronic, Magnetic & Photonic Materials Division, Chemistry & Physics of Materials Committee, Joint Nuclear Materials Committee, and Titanium Committee.     

Microstructure evolution in Zr under equal channel angular pressing

Pure polycrystalline Zr was deformed by equal channel angular pressing (ECAP), and the microstructural characteristics were analyzed. By repeated alternating ECAP, it was possible to, refine the grain size from 200 to 0.2 μm. Subsequent annealing heat treatment at 550°C resulted in a grain growth of up to 6 μm. Mechanical twinning was an important deformation mechanism, particularly during the early stage of deformation. The most active twinning system was identified as 85.2 deg tensile twinning, followed by 57.1 deg compressive twinning. Crystal texture as well as grain-boundary misorientation distribution of deformed Zr were analyzed by X-ray diffraction (XRD) and electron backscattered diffraction (EBSD). The ECAP-deformed Zr showed a considerable difference in the crystallographic attributes from those of cold-rolled Zr or Ti, in that texture and boundary misorientation-angle distribution tend toward more even distribution with a slightly preferential distribution of boundaries of a 20 to 30 deg misorientation angle. Furthermore, unlike the case of cold rolling, the crystal texture was not greatly altered by subsequent annealing heat treatment. Overall, the present work suggests ECAP as a viable method to obtain significant grain refining in hexagonal close-packed (hcp) metals. This article is based on a presentation made in the symposium entitled “Defect Properties and Mechanical Behavior of HCP Metals and Alloys” at the TMS Annual Meeting, February, 11–15, 2001, in New Orleans, Louisiana, under the auspices of the following ASM committees: Materials Science Critical Technology Sector, Structural Materials Division, Electronic, Magnetic & Photonic Materials Division, Chemistry & Physics of Materials Committee, Joint Nuclear Materials Committee, and Titanium Committee.     

Immune response of channel catfish under different environmental conditions

Channel catfish were maintained under conditions of low (15.4 C) and fluctuating (15.4 to 26.9 C/24 hours) temperatures, low (126 mg/L NO3-N) and high (289 mg/L NO3-N) nitrate in recirculating systems, crowding (171 g body mass/L), and fasting. They were vaccinated with formalin-killed enteric red-mouth bacterium, and antibody titers were monitored weekly for 10 weeks. Only those fish maintained in low or fluctuating temperature environments had significant (P less than 0.01) immunosuppression. The other environmental conditions studies, which are commonly encountered in intensive fish culture operations, did not compromise the humoral immune response of channel catfish.     

拉力集中型与压力分散型预应力锚索锚固机理

利用有限差分程序FLAC对一种新型预应力锚索——压力分散型锚索的锚固机理进行了数值模拟研究.与拉力集中型预应力锚索相比,压力分散型锚索具有许多优点,尤其适用于对大变形软岩工程的锚固.对比分析结果表明:压力分散型锚索锚固段浆体轴力峰值仅为拉力集中型锚索的1/n(n为承载体的数量),且浆体处于受压状态;锚固段浆体-岩体界面上的剪力峰值也小于拉力集中型锚索,并沿内锚固段轴向均匀分布;在相同条件下,压力分散型锚索比拉力集中型锚索能提供更大的锚固力;适当增加承载体的数量是提高压力分散型锚索锚固力,改善锚固段浆体受力状态的有效途径。     

Precipitate shape evolution under applied stress—Thermodynamics and kinetics

The influence of an external stress field on the growth and morphological evolution of a coherent precipitate is examined within the context of the thermodynamics of crystalline solids and interfaces. Equilibrium interfacial concentrations are obtained that are shown to be strong functions of position along the precipitate-matrix interface. These variations in concentration may be larger than the usual capillarity term and are a result of elastic considerations and not a result of changes in the local interfacial curvature. The interfacial concentrations directly influence the precipitate growth rate and result in changes in the precipitate morphology. Depending upon the signs and magnitudes of the coherency strains, the applied stress field and the elastic inhomogeneity of the system, the shape evolution may be approximated by ellipsoids of revolution.     

Nucleation kinetics of a solidifying metal under an external action

A mathematical model is developed for the nucleation kinetics of metal solidification under an external action using nonequilibrium thermodynamics. The onset characteristics of nonequilibrium solidification are analyzed. The prenucleus metastable growth time is a function of the prenucleus shape, the physicochemical properties of the melt, and the particle volume. The external action energy intensifies the formation of stable nuclei with sizes smaller than the critical equilibrium size in a shorter metastability period than that during conventional solidification.     

Modeling the reversible kinetics of neutrophil aggregation under hydrodynamic shear

Neutrophil emigration into inflamed tissue is mediated by beta 2-integrin and L-selectin adhesion receptors. Homotypic neutrophil aggregation is also dependent on these molecules, and it provides a model system in which to study adhesion dynamics. In the current study we formulated a mathematical model for cellular aggregation in a linear shear field based on Smoluchowski's two-body collision theory. Neutrophil suspensions activated with chemotactic stimulus and sheared in a cone-plate viscometer rapidly aggregate. Over a range of shear rates (400-800 s-1), approximately 90% of the single cells were recruited into aggregates ranging from doublets to groupings larger than sextuplets. The adhesion efficiency fit to these kinetics reached maximum levels of > 70%. Formed aggregates remained intact and resistant to shear up to 120 s, at which time they spontaneously dissociated back to singlets. The rate of cell disaggregation was linearly proportional to the applied shear rate, and it was approximately 60% lower for doublets as compared to larger aggregates. By accounting for the time-dependent changes in adhesion efficiency, disaggregation rate, and the effects of aggregate geometry, we succeeded in predicting the reversible kinetics of aggregation over a wide range of shear rates and cell concentrations. The combination of viscometry with flow cytometry and mathematical analysis as presented here represents a novel approach to differentiating between the effects of hydrodynamics and the intrinsic biological processes that control cell adhesion.     

Calcium regulation of tension redevelopment kinetics with 2-deoxy-ATP or low [ATP] in rabbit skeletal muscle

The correlation of acto-myosin ATPase rate with tension redevelopment kinetics (k(tr)) was determined during Ca(+2)-activated contractions of demembranated rabbit psoas muscle fibers; the ATPase rate was either increased or decreased relative to control by substitution of ATP (5.0 mM) with 2-deoxy-ATP (dATP) (5.0 mM) or by lowering [ATP] to 0.5 mM, respectively. The activation dependence of k(tr) and unloaded shortening velocity (Vu) was measured with each substrate. With 5.0 mM ATP, Vu depended linearly on tension (P), whereas k(tr) exhibited a nonlinear dependence on P, being relatively independent of P at submaximum levels and rising steeply at P > 0.6-0.7 of maximum tension (Po). With dATP, Vu was 25% greater than control at Po and was elevated at all P > 0.15Po, whereas Po was unchanged. Furthermore, the Ca(+2) sensitivity of both k(tr) and P increased, such that the dependence of k(tr) on P was not significantly different from control, despite an elevation of Vu and maximal k(tr). In contrast, lowering [ATP] caused a slight (8%) elevation of Po, no change in the Ca(+2) sensitivity of P, and a decrease in Vu at all P. Moreover, k(tr) was decreased relative to control at P > 0.75Po, but was elevated at P < 0.75Po. These data demonstrate that the cross-bridge cycling rate dominates k(tr) at maximum but not submaximum levels of Ca(2+) activation.