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Complex-periodic spiral waves in confluent cardiac cell cultures induced by localized inhomogeneities.

Spatiotemporal wave activities in excitable heart tissues have long been the subject of numerous studies because they underlie different forms of cardiac arrhythmias. In particular, understanding the dynamics and the instabilities of spiral waves have become very important because they can cause reentrant tachycardia and their subsequent transitions to fibrillation. Although many aspects of cardiac spiral waves have been investigated through experiments and model simulations, their complex properties are far from well understood. Here, we show that intriguing complex-periodic (such as period-2, period-3, period-4, or aperiodic) spiral wave states can arise in monolayer tissues of cardiac cell culture in vitro, and demonstrate that these different dynamic states can coexist with abrupt and spontaneous transitions among them without any change in system parameters; in other words, the medium supports multistability. Based on extensive image data analysis, we have confirmed that these spiral waves are driven by their tips tracing complex orbits whose unusual, meandering shapes are formed by delicate interplay between localized conduction blocks and nonlinear properties of the culture medium.

Action Potentials↗

Preventing ventricular fibrillation by flattening cardiac restitution.

Ventricular fibrillation is the leading cause of sudden cardiac death. In fibrillation, fragmented electrical waves meander erratically through the heart muscle, creating disordered and ineffective contraction. Theoretical and computer studies, as well as recent experimental evidence, have suggested that fibrillation is created and sustained by the property of restitution of the cardiac action potential duration (that is, its dependence on the previous diastolic interval). The restitution hypothesis states that steeply sloped restitution curves create unstable wave propagation that results in wave break, the event that is necessary for fibrillation. Here we present experimental evidence supporting this idea. In particular, we identify the action of the drug bretylium as a prototype for the future development of effective restitution-based antifibrillatory agents. We show that bretylium acts in accord with the restitution hypothesis: by flattening restitution curves, it prevents wave break and thus prevents fibrillation. It even converts existing fibrillation, either to a periodic state (ventricular tachycardia, which is much more easily controlled) or to quiescent healthy tissue.

Action Potentials↗

Effects of cytochaslasin B and colcemide on myogenic cultures.

Muscle cultures treated with cytochalasin B yield mono- and oligonucleated cells of two kinds: (i) arborized, replicating precursor myogenic cells and fibroblasts; and (ii) round, post-mitotic, terminally differentiating myoblasts and myotubes. The arborized cells do not bind fluorescein-labeled antibody against myosin, do not contract rhythmically, and do not display hexagonally stacked thick and thin filaments. The round, mono-nucleated myoblasts and round, oligonucleated myotubes bind the fluorescein-labeled antibody against myosin, contract rhythmically, and display clusters of hexagonally-stacked thick and thin filaments. When cytochalasin B is removed and replaced by colcemide, the arborized cells, but not the post-mitotic muscle cells, acquire a radial symmetry and are induced to assemble massive, meandering cables that may occupy over 25% of the cell volume. These tortuous calbes are positively birefringent and consist exclusively of enormous numbers of 100-A, intermediate-sized filaments.

Animals↗

Antibodies to liver cell adhesion molecule perturb inductive interactions and alter feather pattern and structure.

Cell adhesion molecules (CAMs) may act as regulators of morphogenesis by constraining cell motion, forming borders, and controlling intercellular communications that lead to embryonic induction. This postulated causal role of CAMs in inductive events was tested here in an in vitro system of feather induction. In the developing chicken skin, an ectodermal sheet of epithelium interacts with mesodermal cell collectives to form more or less circular feather germs arranged in a hexagonal pattern. Cells of the epidermal epithelium are linked by liver CAM (L-CAM) and mesodermal cells in dermal condensations are linked by neural CAM (N-CAM); neither of these CAMs links cells in one tissue of this inductive couple to cells in the other. After perturbation of the L-CAM linkage in epidermis by antibodies to L-CAM, nonhexagonal striped patterns of dermal condensations were observed in culture. The stripes did not follow straight lines but meandered in lateral and oblique directions. Histological examination of the perturbed tissues showed extensive changes in dermal cell density distributions. After 10 days of culture, the perturbed tissues developed a cobbled or plaque-like morphology resembling scales rather than the feather-like filamentous structures that formed in unperturbed skin cultures. The results indicate that perturbation of CAM binding in tissues linked by one CAM can alter fates and interactions of cells linked by another, presumably by altering the amount or effect of inductive signals crossing the border between the inducing cell collectives. A computer model based on the notion that the response of L-CAM-linked epidermal cells to signals from N-CAM-linked dermal cells depends cooperatively on the degree of L-CAM linkage was found to generate hexagonal patterns for the unperturbed case and stripes after perturbation of L-CAM bonds.

Animals↗

Three-dimensional structure of human basic fibroblast growth factor, a structural homolog of interleukin 1 beta.

The three-dimensional structure of the 146-residue form of human basic fibroblast growth factor (bFGF), expressed as a recombinant protein in yeast, has been determined by x-ray crystallography to a resolution of 1.8 A. bFGF is composed entirely of beta-sheet structure, comprising a three-fold repeat of a four-stranded antiparallel beta-meander. The topology of bFGF is identical to that of interleukin 1 beta, showing that although the two proteins share only 10% sequence identity, bFGF, interleukin 1, and their homologs comprise a family of structurally related mitogenic factors. Analysis of the three-dimensional structure in light of functional studies of bFGF suggests that the receptor binding site and the positively charged heparin binding site correspond to adjacent but separate loci on the beta-barrel.

Amino Acid Sequence↗

Three-dimensional structure of human protein kinase C interacting protein 1, a member of the HIT family of proteins.

The three-dimensional structure of protein kinase C interacting protein 1 (PKCI-1) has been solved to high resolution by x-ray crystallography using single isomorphous replacement with anomalous scattering. The gene encoding human PKCI-1 was cloned from a cDNA library by using a partial sequence obtained from interactions identified in the yeast two-hybrid system between PKCI-1 and the regulatory domain of protein kinase C-beta. The PKCI-1 protein was expressed in Pichia pastoris as a dimer of two 13.7-kDa polypeptides. PKCI-1 is a member of the HIT family of proteins, shown by sequence identity to be conserved in a broad range of organisms including mycoplasma, plants, and humans. Despite the ubiquity of this protein sequence in nature, no distinct function has been shown for the protein product in vitro or in vivo. The PKCI-1 protomer has an alpha+beta meander fold containing a five-stranded antiparallel sheet and two helices. Two protomers come together to form a 10-stranded antiparallel sheet with extensive contacts between a helix and carboxy terminal amino acids of a protomer with the corresponding amino acids in the other protomer. PKCI-1 has been shown to interact specifically with zinc. The three-dimensional structure has been solved in the presence and absence of zinc and in two crystal forms. The structure of human PKCI-1 provides a model of this family of proteins which suggests a stable fold conserved throughout nature.

Amino Acid Sequence↗

Corrosion casts of convoluted testicular arteries in mice and rats.

The preparation of vascular corrosion casts enables us to observe the minute features of blood vessels. For mice and rats, reports on the macroscopical appearance of testicular arteries with the cast technique have been quite limited. Therefore, in the present study, we tried to prepare corrosion casts of such arteries in order to observe the three-dimensional structures in mice and rats. Vascular casts of testicular arteries were produced by intravenous injection of a polymethylmethacrylate solution, followed by treatment of the whole animal bodies with NaOH. Additionally, the gross appearance of human testicular arteries in cadavers was compared with that of the vascular casts. The results showed that the testicular arteries of humans, mice, and rats run from the abdominal aorta to the testes, with various configurations (straight, meandering, spiral, or coiled forms), and each species exhibits a specific pattern. The specific forms of testicular arteries may play several roles in protection of normal spermatogenesis, such as allowing wide mobility of the testes on physical attack, heat emission with the entwined pampiniform plexus, and reduction of the blood flow rate. The great extension of the testicular arteries during the fetal period from the upper abdomen to the scrotum when the testes descend might be another feature of the specific development of their running configuration.

Animals↗

Molecular dynamics simulations of beta-turn forming tetra- and hexapeptides.

It was previously shown that the structural ensemble of model peptides DDKG and GKDG (H. Ishii et al. Biopolymers 24, 2045-2056, 1985), DEKS (A. Otter et al. J. Biomol. Struct. Dyn. 7, 455-476, 1989) NPGQ (F. R. Carbone et al. Int. J. Pept. Protein. Res. 26, 498-508, 1985), SALN (H. Santa et al. J. Biomol. Struct. Dyn. 16, 1033-1041, 1999), SYPFDV and SYPYDV (J. Yao et al. J. Mol. Biol. 243, 736-753, 1994), VP(D)AH and VP(D)SH (B. Imperiali et al. J. Am. Chem. Soc. 114, 3182-3188, 1992) in solution contains a significant - or in some cases dominant - proportion of beta-turn conformation. In this study, a protein database was searched for the above, unprotected sequences which incorporate only L-amino acid residues. Simulated annealing and 25 ns MD simulations of structures were also performed. The DSSP and STRIDE secondary structure-assigning algorithms and clustering were used to analyze trajectories and i, i+3 hydrogen bonds were also sought. The DSSP analysis showed a fluctuation between beta-turn and random meander structure, although bend structures were not detected because of the insufficient length of peptide chains. This alternating trend was confirmed when the STRIDE algorithm was used to analyze trajectories, but STRIDE assigned more turn structures. The population of the strongest clusters was above 40% and the middle structures adopted beta-turn structure for most sequences. These results are in good agreement with previous experimental results and support the idea of the ultra-marginal stability of turns in the absence of stabilizing long-range interactions of the neighboring segments of a polypeptide chain. However, interactions between the side-chains in tetrapeptides could also contribute to turn stability and result in unusual stability in some cases. Our observations suggest that such interactions are the consequence rather than the driving force of turn formation.

Amino Acid Sequence↗

Meteorological factors of ozone predictability at Houston, Texas.

Several ozone modeling approaches were investigated to determine if uncertainties in the meteorological data would be sufficiently large to limit the application of physically realistic ozone (O3) forecast models. Three diagnostic schemes were evaluated for the period of May through September 1997 for Houston, TX. Correlations between measured daily maximum and model calculated O3 air concentrations were found to be 0.70 using a linear regression model, 0.65 using a non-advective box model, and 0.49 using a three-dimensional (3-D) transport and dispersion model. Although the regression model had the highest correlation, it showed substantial underestimates of the highest concentrations. The box model results were the most similar to the regression model and did not show as much underestimation. The more complex 3-D modeling approach yielded the worst results, likely resulting from O3 maxima that were driven by local factors rather than by the transport of pollutants from outside of the Houston domain. The highest O3 concentrations at Houston were associated with light winds and meandering trajectories. A comparison of the gridded meteorological data used by the 3-D model to the observations showed that the wind direction and speed values at Houston differed most on those days on which the O3 underestimations were the greatest. These periods also tended to correspond with poor precipitation and temperature estimates. It is concluded that better results are not just obtained through additional modeling complexity, but there needs to be a comparable increase in the accuracy of the meteorological data.

Air Pollution↗

Three-dimensional modeling of air flow and pollutant dispersion in an urban street canyon with thermal effects.

Effects of excess ground and building temperatures on airflow and dispersion of pollutants in an urban street canyon with an aspect ratio of 0.8 and a length-to-width ratio of 3 were investigated numerically. Three-dimensional governing equations of mass, momentum, energy, and species were modeled using the RNG k-epsilon turbulence model and Boussinesq approximation, which were solved using the finite volume method. Vehicle emissions were estimated from the measured traffic flow rates and modeled as banded line sources, with a street length and bandwidths equal to typical vehicle widths. Both measurements and simulations reveal that pollutant concentrations typically follow the traffic flow rate; they decline as the height increases and are higher on the leeward side than on the windward side. Three-dimensional simulations reveal that the vortex line, joining the centers of cross-sectional vortexes of the street canyon, meanders between street buildings and shifts toward the windward side when heating strength is increased. Thermal boundary layers are very thin. Entrainment of outside air increases, and pollutant concentration decreases with increasing heating condition. Also, traffic-produced turbulence enhances the turbulent kinetic energy and the mixing of temperature and admixtures in the canyon. Factors affecting the inaccuracy of the simulations are addressed.

Air Movements↗

The cytoskeletal and contractile apparatus of smooth muscle: contraction bands and segmentation of the contractile elements.

Confocal laser scanning microscopy of isolated and antibody-labeled avian gizzard smooth muscle cells has revealed the global organization of the contractile and cytoskeletal elements. The cytoskeleton, marked by antibodies to desmin and filamin is composed of a mainly longitudinal, meandering and branched system of fibrils that contrasts with the plait-like, interdigitating arrangement of linear fibrils of the contractile apparatus, labeled with antibodies to myosin and tropomyosin. Although desmin and filamin were colocalized in the body of the cell, filamin antibodies labeled additionally the vinculin-containing surface plaques. In confocal optical sections the contractile fibrils showed a continuous label for myosin for at least 5 microns along their length: there was no obvious or regular interruption of label as might be expected for registered myosin filaments. The cytoplasmic dense bodies, labeled with antibodies to alpha-actinin exhibited a regular, diagonal arrangement in both extended cells and in cells shortened in solution to one-fifth of their extended length: after the same shortening, the fibrils of the cytoskeleton that showed colocalization with the dense bodies in extended cells became crumpled and disordered. It is concluded that the dense bodies serve as coupling elements between the cytoskeletal and contractile systems. After extraction with Triton X-100, isolated cells bound so firmly to a glass substrate that they were unable to shorten as a whole when exposed to exogenous Mg ATP. Instead, they contracted internally, producing integral of 10 regularly spaced contraction nodes along their length. On the basis of differences of actin distribution two types of nodes could be distinguished: actin-positive nodes, in which actin straddled the node, and actin-negative nodes, characterized by an actin-free center flanked by actin fringes of 4.5 microns minimum length on either side. Myosin was concentrated in the center of the node in both cases. The differences in node morphology could be correlated with different degrees of coupling of the contractile with the cytoskeletal elements, effected by a preparation-dependent variability of proteolysis of the cells. The nodes were shown to be closely related to the supercontracted cell fragments shown in the accompanying paper (Small et al., 1990) and furnished further evidence for long actin filaments in smooth muscle. Further, the segmentation of the contractile elements pointed to a hierarchial organization of the myofilaments governed by as yet undetected elements.

Actinin↗

Electron microscopy of peripheral nerves and neuromuscular junctions in the wasp leg.

The peripheral nerve branch innervating the femoral muscles of the common yellow jacket (Vespula carolina) has been found to possess a thick lemnoblast basement membrane and a complex mesaxon. The term "tunicated nerve" is proposed to designate the type of peripheral nerve in which one or several axons are loosely mantled by meandering, cytoplasm-enclosing membranes of the lemnoblast. The peripheral axon courses longitudinally in a groove in the muscle fiber between the plasma membrane of the muscle fiber and a cap formed by lemnoblast and tracheoblast. The junction is characterized by apposition of plasma membranes of axon and muscle fiber, abundant mitochondria, and synaptic vesicles in the axon, and aggregates of "aposynaptic granules" plus mitochondria and endoplasmic reticulum on the muscle side of the synapse. Unlike the vertebrate striated muscle fiber, no complex infolding of the synapsing plasma membrane of the muscle fiber occurs. The "connecting tissue" of the insect is formed by tracheoblasts, their basement membranes, and the basement membranes of other cells. Further mechanical support is given by the ramifying tracheoles. The physiologic roles of the specialized structures are considered.

Animals↗

Neuronal perikarya with dispersed, single ribosomes in the visual cortex of Macaca mulatta.

Numerous small and medium-sized neuronal perikarya in layers III and IV of the visual cortex display an unusual pattern of ribosomal distribution. Instead of being aggregated in clusters, spirals, rows, and other regular polysomal configurations, the ribosomes, whether free or attached to the endoplasmic reticulum, are randomly dispersed, with no discernible pattern. The endoplasmic reticulum in such cells is reduced to a few (perhaps only one) meandering, broad cisternae, which delimit broad fields of cytoplasmic matrix occupied almost solely by scattered, single ribosomes. The Golgi apparatus is elaborate. Mitochondria are either small and numerous or large and infrequent. The other organelles, including the nucleus and nucleolus, are not remarkable. Axonal terminals synapse in the normal fashion on the surfaces of these cells and their dendrites. Associated with these cells are more numerous intermediate cells in which a few to many polysomal clusters can be found. It is proposed that the neurons with dispersed, single ribosomes are inactive in protein synthesis and that the suspension of such an important metabolic activity is probably temporary. Thus, these cells are considered to be part of a population undergoing cyclic fluctuations in the intensity of protein synthesis that should be correlated with their specific neural behavior.

Animals↗

Size and shape of transverse tubule openings in frog twitch muscle fibers.

The openings of transverse tubules in frog twitch fibers are described. The tubules open to the extracellular space by a narrow neck, with an inner diameter of 20 nm. The most peripheral portion of the tubules is tortuous and has a variable diameter. The similarity in size of the openings of T tubules and caveolae and the meandering path of the tubules are sufficient to account for the paucity of observed openings.

Animals↗

The fine structure of Meissner's touch corpuscles of human fingers.

Thin slices of the finger pads of six individuals were fixed in buffered 1 per cent osmic acid, embedded in deaerated, nitrogenated methacrylate, and cut into thin sections for electron microscopic study. Before embedding, the slices were trimmed so as to include several digital tactile corpuscles. Some thin sections were stained in 10 per cent aqueous phosphotungstic acid solution. The principal part of Meissner's corpuscle is made up of flattened laminar cells stretching across the corpuscle in irregular layers. The perinuclear cytoplasm of these cells contains numerous small mitochondria, a sparse granular endoplasmic reticulum, and a large number of small vesicles. Nerve fibers enter the side or base of the corpuscle, lose their myelin sheaths, and follow a meandering course between the laminar cell plates. The nerve endings enter into a close appositional relationship with the flattened portions of the laminar cells. In some areas the apposed axolemma and cell membranes are slightly thickened with small vesicles located along the cell membrane or on both surfaces. These regions are interpreted as synapses. The most prominent feature of the nerve endings is an extraordinary accumulation of small mitochondria which vary in size and internal density. The nerve endings also contain vacuoles, groups of dense concentric membranes, and small dense vesicles of irregular distribution. The laminar cells are separated from one another by a dense intercellular substance of uniform thickness which also envelops the entire corpuscle. This material contains randomly oriented collagen fibers and fine fibrils bound together by a dense material at nodal points recurring at regular intervals of approximately 120 mmicro. These findings are discussed in relation to the problems of the function of Meissner's corpuscle, neural material loss and replacement, and the presence of synapses.

Animals↗

Biogeography of luminous marine ostracod driven irreversibly by the Japan current.

The biogeography of the luminous marine ostracod Vargula hilgendorfii, also called "Umihotaru," shows that this organism may have arrived relatively recently on the Japanese islands during the final glacier period approximately 10,000 years ago. Phylogenetic relationships also strongly indicate that the Japan Current drove the Umihotaru ostracod northward. It is evident that the Umihotaru ostracod spread rapidly to the major Japanese islands 3,000 km north, whereas its spread was slow in the southwest of the Japanese islands, covering a distance of 400 km. The meandering of the Japan Current, where it passes by the Tokara Gap at 28 degrees N latitude, may be a barrier to Umihotaru ostracod extension.

Animals↗

MICROANATOMY, ULTRASTRUCTURE, AND SYSTEMATIC SIGNIFICANCE OF THE EXCRETORY SYSTEM AND MANTLE CAVITY OF AN ACOCHLIDIAN GASTROPOD (OPISTHOBRANCHIA).

The microanatomy and ultrastructure of the excretory system of an undescribed mesopsammic gastropod of the genus Hedylopsis have been examined by means of semithin serial sections, reconstructions, and transmission electron microscopy. The functional metanephridial system comprises a monotocardian heart with a single ventricle and auricle in a spacious pericardium as well as a single, large kidney. Podocytes in the auricular epicardium represent the site of ultrafiltration and formation of the primary urine, whereas the flat epithelium of the kidney with extensive basal infoldings, large vacuoles and the apical microvillous border indicates modification of the primary filtrate. Solitary rhogocytes (pore cells) represent additional loci of ultrafiltration with an identical fine-structure as those of the podocytes (meandering slits with diaphragms covered by extracellular matrix). The presence of podocytes situated in the epicardial wall of the auricle is regarded as plesiomorphic for the Opisthobranchia and is confirmed for the Acochlidia for the first time. Kidney and rectum both open into a small, yet distinct mantle cavity. Within the Acochlidia this condition represents a plesiomorphic character only known from one further Hedylopsis species until now. Special cells (here termed microvillous pit-cells) with a presumed absorptive function are interspersed between the epithelial cells of the mantle cavity. They are mainly characterized by a prominent invagination of the apical border with densely arranged, very large microvilli. The presence of a mantle cavity that has been lost in all other acochlidian genera supports the systematic placement of the Hedylopsidae at the base of the Achochlidia.

Journal Article↗

Electron microscopy analysis of the boundary layer structure of SrTiO3 semiconducting ceramic

In a boundary layer (BL) semiconducting SrTiO3-based ceramic condenser, the BL structure has been investigated using high-resolution field-emission scanning electron microscopy (FE-SEM) and field-emission (scanning) transmission electron microscopy (FE-(S)TEM). In an initial TEM observation, a double layered structure was observed at the grain boundary region. It consisted of a grain boundary (second phase) and a pair of the metal diffusion layers of up to several nanometres in width across the grain boundary where the change of the crystal lattice distance was undetected by the high-resolution TEM image. A facet structure was often observed on the grain boundaries. It was particularly formed on (020) plane of the grain crystal. High resolution SEM showed a jagged striped structure on the surface of the bulk material and on the inside grain as revealed by fracture. Using the similarity in shape and size, it can be identified to correspond to the facet boundary structure. Its formation mechanism can be explained as that during the reoxidization process when the oxide flux of the mixture of Bi2O3, PbO and CuO, painted on the bulk material, surfaces migrates into the ceramic along the grain boundary. The oxide corrodes the grain surfaces including the bulk surfaces. This corrosion particularly occurs on (020) plane of the grain so that the facet structure is produced. In this paper, by using the atomic scale high angle annular dark field STEM, it has been determined that Bi atoms preferentially replace Sr atoms on (020) in the diffusion layers. The atom position displacement was also detected at the grain surfaces and this altered atomic assignment can be determined as an origin of production of Sr2Bi4Ti5O18 at the grain boundary. Also, it was observed that the layer width of the metal diffusion layers was often different between the both grains and changed locally so that the ribbon of the diffusion layers meandered around the straight grain boundary. Its possible mechanism is also proposed.

Journal Article↗