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Airway smooth muscle is able to adapt and maintain a nearly constant maximal force generation over a large length range. This implies that a fixed filament lattice such as that found in striated muscle may not exist in this tissue and that plastic remodeling of its contractile and cytoskeletal filaments may be involved in the process of length adaptation that optimizes contractile filament overlap. Here, we show that isometric force produced by airway smooth muscle is independent of muscle length over a twofold length change; cell cross-sectional area was inversely proportional to cell length, implying that the cell volume was conserved at different lengths; shortening velocity and myosin filament density varied similarly to length change: increased by 69.4% +/- 5.7 (SE) and 76.0% +/- 9.8, respectively, for a 100% increase in cell length. Muscle power output, ATPase rate, and myosin filament density also have the same dependence on muscle cell length: increased by 35.4% +/- 6.7, 34.6% +/- 3.4, and 35.6% +/- 10.6, respectively, for a 50% increase in cell length. The data can be explained by a model in which additional contractile units containing myosin filaments are formed and placed in series with existing contractile units when the muscle is adapted at a longer length.

We investigated the role of mitochondria (MT) in calcium signaling in a culture of rat aortic smooth muscle cells. We used targeted aequorin to selectively measure [Ca2+] in this organelle. Our results reveal that smooth muscle cell stimulation with agonists causes a large, transient increase in mitochondrial [Ca2+] ([Ca2+]m). This large transient can be blocked with inhibitors of the sarco-endoplasmic reticulum Ca2+-ATPase, suggesting a close relationship between the sarcoplasmic reticulum (SR) and the mitochondria. FCCP completely abolished the response to agonists, and targeted mitochondrial GFP revealed a vast tubular network of MT in these cells. When added before stimulation with ATP, IP3 inhibitors partially blocked the ATP-induced rise in mitochondrial Ca2+ release. The role of the Na+/Ca2+ exchanger (NCX) was examined by removing extracellular Na+. This procedure prevented the decrease in the [Ca2+]m transient normally seen on removal of extracellular Ca2+. We propose a functional linkage of MT and SR dependent on a narrow junctional space between the two organelles in which Ca2+ diffusion is restricted. Approximately half of the mitochondria appear to be associated with the superficial SR, which communicates with the extracellular space via NCX.

Phosphorylation of the 20-kDa regulatory myosin light chain (MLC) of smooth muscle is known to cause monomeric myosins in solution to self-assemble into thick filaments. The role of MLC phosphorylation in thick filament formation in intact muscle, however, is not clear. It is not known whether the phosphorylation is necessary to initiate thick filament assembly in vivo. Here we show, by using a potent inhibitor of MLC kinase (wortmannin), that the MLC phosphorylation and isometric force in trachealis muscle could be abolished without affecting calcium transients. By measuring cross-sectional densities of the thick filaments electron microscopically, we also show that inhibition of MLC phosphorylation alone did not cause disassembly of the filaments. The unphosphorylated thick filaments, however, partially dissolved when the muscle was subjected to oscillatory strains (which caused a 25% decrease in the thick filament density). The postoscillation filament density recovered to the preoscillation level only when wortmannin was removed and the muscle was stimulated. The data suggest that in vivo thick filament reassembly after mechanical perturbation is facilitated by the cyclic MLC phosphorylation associated with repeated stimulation.

Vascular smooth muscle shows both plasticity and heterogeneity with respect to Ca(2+) signaling. Physiological perturbations in cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) may take the form of a uniform maintained rise, a transient uniform [Ca(2+)](i) elevation, a transient localized rise in [Ca(2+)](i) (also known as spark and puff), a transient propagated wave of localized [Ca(2+)](i) elevation (Ca(2+) wave), recurring asynchronous Ca(2+) waves, or recurring synchronized Ca(2+) waves dependent on the type of blood vessel and the nature of stimulation. In this overview, evidence is presented which demonstrates that interactions of ion transporters located in the membranes of the cell, sarcoplasmic reticulum, and mitochondria form the basis of this plasticity of Ca(2+) signaling. We focus in particular on how the junctional complexes of plasmalemma and superficial sarcoplasmic reticulum, through the generation of local cytoplasmic Ca(2+) gradients, maintain [Ca(2+)](i) oscillations, couple these to either contraction or relaxation, and promote Ca(2+) cycling during homeostasis.

Myosin thick filaments have been shown to be structurally labile in intact smooth muscles. Although the mechanism of thick filament assembly/disassembly for purified myosins in solution has been well described, regulation of thick filament formation in intact muscle is still poorly understood. The present study investigates the effect of resting calcium level on thick filament maintenance in intact airway smooth muscle and on thick filament formation during activation. Cross-sectional density of the thick filaments measured electron microscopically showed that the density increased substantially (144%) when the muscle was activated. The abundance of filamentous myosins in relaxed muscle was calcium sensitive; in the absence of calcium (with EGTA), the filament density deceased by 35%. Length oscillation imposed on the muscle under zero-calcium conditions produced no further reduction in the density. Isometric force and filament density recovered fully after reincubation of the muscle in normal physiological saline. The results suggest that in airway smooth muscle, filamentous myosins exist in equilibrium with monomeric myosins; muscle activation favors filament formation, and the resting calcium level is crucial for preservation of the filaments in the relaxed state.

Ionic interactions between the plasma membrane (PM) and the sarcoplasmic reticulum (SR) play a crucial role in smooth muscle activation and homeostasis. The most common form of Ca2+ signalling seen in vascular smooth musde of conduit arteries and capacitance veins consists of repetitive asynchronous Ca2+ waves. In the inferior vena cava of the rabbit these waves are initiated by Ca2+ release via InsP3 receptors (InsP3R) and propagated by regenerative Ca2+ release. Maintenance of the [Ca2+] oscillations is dependent on Ca2+ entry through the Na+/Ca2+-exchanger (NCX) which is driven in the reverse mode by Na+ entry through non-specific cation channels. The latter are also responsible for depolarization and activation of voltage-gated Ca2+ channels. The sarcoplasmic-endoplasmic reticulum Ca2+-ATPase (SERCA) in the sheet-like junctional SR is responsible for refilling and completing the cycle. Under resting conditions the interaction between the superficial SR and the NCX is reversed with Ca2+ release channels supplying Ca2+ to the NCX in the PM to be extruded in exchange of extracellular Na+. It is proposed that the above Ca2+ transport between the SR lumen and the extracellular space takes place at PM-SR junctions across a narrow junctional space.

In the present study, the immersion behavior of two kinds of sintered HA with different Ca/P ratios in two different extracellular simulated solutions (Tris buffer and Hank's solutions) was investigated and compared. Results indicated that an as-received Ca-deficient HA (FHA) had a lower Ca/P ratio, larger linear shrinkage and higher density than a stoichiometric HA (MHA). When FHA powder was calcined at 900 degrees C, its Ca-deficient apatite structure was unstable and a significant amount of beta-TCP phase was formed. When heated to 1250 degrees C in air, the highly crystalline apatite structure of MHA was still stable without any noticeable decomposition. The FTIR spectra indicated that both immersed MHA and FHA in Hank's solution were gradually covered with a layer of precipitated apatite during immersion. When immersed in Tris buffer solution, neither HA showed significant changes in their FTIR spectra. SEM observation indicated that the precipitation rate on immersed FHA surface was much higher than that on MHA surface when immersed in Hank's solution. The weight loss and pH data confirmed the higher dissolution rate of FHA than MHA in Hank's solution., (Copyright 2001 Kluwer Academic Publishers)

Airway smooth muscle adapts to different lengths with functional changes that suggest plastic alterations in the filament lattice. To look for structural changes that might be associated with this plasticity, we studied the relationship between isometric force generation and myosin thick filament density in cell cross sections, measured by electron microscope, after length oscillations applied to the relaxed porcine trachealis muscle. Muscles were stimulated regularly for 12 s every 5 min. Between two stimulations, the muscles were submitted to repeated passive +/- 30% length changes. This caused tetanic force and thick-filament density to fall by 21 and 27%, respectively. However, in subsequent tetani, both force and filament density recovered to preoscillation levels. These findings indicate that thick filaments in airway smooth muscle are labile, depolymerization of the myosin filaments can be induced by mechanical strain, and repolymerization of the thick filaments underlies force recovery after the oscillation. This thick-filament lability would greatly facilitate plastic changes of lattice length and explain why airway smooth muscle is able to function over a large length range.

The base-centered orthorhombic approximant Ga7Mn5 (a = 4.547(6) nm, b = 1.256(9) nm, c = 1.482(1) nm) consisting of flattened hexagon and concave octagon (called "crown" in this paper) subunits shown by high-resolution electron microscopy (HREM) was found to coexist with the Ga-Mn decagonal quasicrystal in the Ga58Mn42 alloy. The atomic sites in the flattened hexagon subunit have been deduced recently from a structural study of the orthorhombic Ga-Mn (1/1,1/1) approximant that consists only of the hexagon subunits. Similarly, the atomic sites in the crown subunit have been deduced from another orthorhombic Ga-Mn (2/1,1/1) approximant consisting only of the crown subunits. A structural model of the orthorhombic Ga7Mn5 can thus be constructed from these structural subunits. There are 332 Ga atoms and 232 Mn atoms in a unit cell, with a composition of Mn41.2Ga58.8, satisfying the symmetry of the space group Bmm2. The simulated electron diffraction patterns, [010] HREM images, and X-ray powder diffraction pattern based on this structural model agrees mainly with the experimental observations.

Nu-Al(80.61)Cr(10.71)Fe(8.68), P6(3)/m (No. 176), a = 40.68 (7), c = 12.546 (1) A, V = 17,983 (8) A3, atoms/cell = 1184.56, Dx = 3.518 g cm(-3), lambda(Mo K alpha) = 0.71069 A, mu = 5.032 mm(-1), F(000) = 18,433, T = 293 K, final R = 0.075 for 3854 reflections with Fo > 4sigma(Fo). The [001] high-resolution electron-microscopic image of the nu-AlCrFe phase clearly shows similar local characteristics to those given by the complex icosahedral cluster found in somewhat smaller hexagonal approximant structures, such as kappa-Al76Cr18Ni6 [a = 17.674 (3), c = 12.516 (3) A; Sato et al. (1997). Acta Cryst. C53, 1531-1533; Marsh (1998). Acta Cryst. B54, 925-926] and lambda-Al(4.32)Mn [a = 28.382 (9), c = 12.389 (2) A; Kreiner & Franzen (1997). J. Alloys Compd. 261, 83-104]. Using the known atomic distribution of this icosahedral cluster in the kappa and lambda phases as the starting point, the structure of the nu phase, a hexagonal intermetallic compound with probably the largest a parameter, was solved by X-ray single-crystal diffraction using direct methods. As in kappa and lambda phases, almost all TM (transition metal) atoms in the complex icosahedral cluster are icosahedrally coordinated. However, contrary to the lambda structure in which about 98% of the TM atoms have icosahedral-coordination, the TM atoms in the v structure also form capped pentagonal prisms in the region between these complex icosahedral clusters, yielding an average icosahedral coordination of about 70% for TM atoms. After rapid solidification, the v phase occurs together with a decagonal quasicrystal with a periodicity of about 12.5 A along its tenfold axis and thus also consists of six layers, two flat ones each sandwiched between two puckered layers in mirror reflection, stacked along the c axis.

The electrical activity of four pairs of synergistic muscles-the long head of triceps brachii and anconeus, lateral gastrocnemius and soleus, the long and short heads of biceps brachii, and rectus femoris and vastus medialis-was studied during isometric contractions of varying speed. Each muscle pair consists of a two-joint muscle and a one-joint muscle. Two of the one-joint muscles, anconeus and soleus, are composed predominantly of red muscle fibers and are called red muscles; all the remaining muscles are composed predominantly of pale fibers and are therefore called pale muscles. Synergistic pairs of muscles in which the muscles are both pale become active simultaneously in movement of all speeds. No difference in usage of such muscles was found as a function of movement speed. In synergistic muscle pairs composed of a red and a pale muscle, a slow movement was always initiated by the red muscle, whose electrical activity predominated throughout the movement. In a rapid movement, the pale muscle could be initially most active. However, movements of equal speed could be initiated by the red muscle. Thus the speed of a contraction is a necessary but not a sufficient condition for reversals of muscle activation.

The compound 4,5,6,7-tetrachloro-2-methylbenzimidazole (TMB), has been found to markedly modify the steady-state valinomycin-mediated conductance of potassium (K+) ions through lipid bilayer membranes. TMB alone does not contribute significantly to membrane conductance, being electrically neutral in solution. In one of two classes of experiments (I), valinomycin is first added to the aqueous phases, then changes of membrane conductance accompanying stepwise addition of TMB to the water are measured. In a second class of experiments (II), valinomycin is added to the membrane-forming solution, followed by TMB additions to the surrounding water. In both cases membrane conductance shows an initial increase with increasing TMB concentration which is more pronounced at lower K+ ion concentration. At TMB concentrations in excess of 10(-5) M, membrane conductance becomes independent of K+ ion concentration, in contrast to the linear dependence observed at TMB concentrations below 10(-7) M. This transition is accompanied by a change of high field current-voltage characteristics from superlinear (or weakly sublinear) to a strongly sublinear form. All of these observations may be correlated by the kinetic model for carrier-mediated transport proposed by Läuger and Stark (Biochim. Biophys. Acta 211:458, 1970) from which it may be concluded that valinomycin-mediated ion transport is limited by back diffusion of the uncomplexed carrier at high TMB concentrations. Experiments of class I reveal a sharp drop of conductance at high (greater than 10(-5) M) TMB concentration, not seen in class II experiments, which is attributed to blocked entry of uncomplexed carrier from the aqueous phases. Valinomycin initially in the membrane is removed by lateral diffusion to the surrounding torus. The time dependence of this removal has been studied in a separate series of experiments, leading to a measured coefficient of lateral diffusion for valinomycin of 5 x 10(-6) cm2/sec at 25 degrees C. This value is about two orders of magnitude larger than the corresponding coefficient for transmembrane carrier diffusion, and provides further evidence for localization of valinomycin in the membrane/solution interfaces.

Valinomycin selectively transports alkali cations, e.g. potassium ions, across lipid bilayer membranes. The blocking of this carrier-mediated transport by four substituted benzimidazoles has been investigated. The compounds are 4,5,6,7-tetrachloro-2-trifluoromethylbenzimidazole, (TTFB); 4,5,6,7,-tetrachloro-2-methylbenzimidazole, (TMB); 2-trifluoromethylbenzimidazole, (TFB); and 2-methylbenzimidazole, (MBM). Because of its low acidic dissociation constant (pKa = 5.04), the blocking efficiency of TTFB in both neutral and anionic forms in the aqueous phase could be studied. The compounds exhibit the blocking efficiency sequence, TTFB- greater than TTFB0 greater than TMB0 greater than TFB0 greater than MBM0. The corresponding scale of decreasing lipophilicity, as determined by octanol/water partitioning, is TTFB0 greater than TMB0 greater than TTFB- greater than TFB0 greater than MBM0. Comparison of neutral species establishes a positive correlation of blocking efficiency with lipophilicity, with the latter being conferred primarily by chlorination of the benzenoid nucleus. Anionic TTFB, on the other hand, is the most effective blocking agent studied in spite of the fact that its dissociation in the aqueous phase markedly impedes its entry (presumably as a neutral species) into a bulk hydrocarbon phase. This observation suggests that the blocking of valinomycin-mediated bilayer membrane conductance takes place at the membrane/solution interface.

High-resolution electron microscopy has been used to characterize the platinum particles supported on TiO2 or ZnO. After reduction at elevated temperatures, the metallic particles display a regular, faceted shape, and several superstructures, Pt3 Ti(C), Pt3 Ti(H), PtTi, and PtZn, have been found. These results, which may involve strong metal-support interaction, have been confirmed by optical diffraction and image simulation.

The crystal imperfections in thin films of lanthanide phthalocyanines (LnPc2H, Ln = Nd, Tb, Er, Tm, Yb, and Lu) grown expitaxially on KCl have been observed by molecular imaging. Grain and twin boundaries, stacking faults, point defects, vacancies, mosaic structures, and sometimes even some amorphous islands exist in the well-crystallized specimens. Combined with the results reported earlier, the packing characteristics of planar LnPc2H molecules can be well understood.

New findings on quasicrystals with icosahedral, octagonal, decagonal, and dodecagonal symmetries obtained recently in the Beijing Laboratory of Electron Microscopy, Chinese Academy of Sciences, are presented. Special emphasis is put on the relation between quasicrystalline and crystalline structures. The important role played by electron diffraction and high-resolution electron microscopy in revealing these quasiperiodic structures is pointed out.

Epitaxial films of lanthanide phthalocyanines (LnPc2H, Ln = Nd, Tb, Er, Tm, Yb, or Lu) formed on a KCl or NaCl crystal have been studied by means of high-resolution electron microscopy. These complexes are isomorphous and have both tetragonal and base-centered orthorhombic structures. Though radiation damage makes it difficult to study these beam-sensitive materials by electron microscopy, the behaviors of thin film growth and details of structural defects can still be identified at the molecular level.