Changes in host and parasite-derived cellular and extracellular matrix components in developing cysts of Myxobolus pendula (Myxozoa)

Cysts of Myxobolus pendula from the gill arch of creek chub (Semotilus atromaculatus) were examined at various stages of development using light and electron microscopy. The subepithelial host connective tissue underwent dramatic changes, including degradation and remodelling of collagen and vascularisation, in response to the infection. Inflammatory cells lay in a fluid-filled space beneath the host's connective tissue and surrounded a distinctive parasite-derived matrix, composed of collagen fibril bundles embedded in cellular processes of the underlying secretory cells. This collagen matrix was resistant to degradation and invasion by leukocytes. Secretion of a matrix by M. pendula as a structural support, and a protective barrier against the host inflammatory cells is a novel observation for cyst-forming Myxosporea.     

AICAR and phlorizin reverse the hypoglycemia-specific defect in glucagon secretion in the diabetic BB rat

Individuals with type 1 diabetes demonstrate a hypoglycemia-specific defect in glucagon secretion. To determine whether intraislet hyperinsulinemia plays a role in the genesis of this defect, glucagon-secretory responses to moderate hypoglycemia induced by either insulin or a novel combination of the noninsulin glucose-lowering agents 5-aminoimidazole-4-carboxamide (AICAR) and phlorizin were compared in diabetic BB rats (an animal model of type 1 diabetes) and nondiabetic BB rats. The phlorizin-AICAR combination was able to induce moderate and equivalent hypoglycemia in both diabetic and nondiabetic BB rats in the absence of marked hyperinsulinemia. Diabetic BB rats demonstrated impaired glucagon and epinephrine responses during insulin-induced hypoglycemia compared with nondiabetic rats. In contrast, both glucagon (9- to 10-fold increase) and epinephrine (5- to 6-fold increase) responses were markedly improved during phlorizin-AICAR hypoglycemia. Combining phlorizin, AICAR, and insulin attenuated the glucagon response to hypoglycemia by 70% in the diabetic BB rat. Phlorizin plus AICAR had no effect on counterregulatory hormones under euglycemic conditions. We conclude that alpha-cell glucagon secretion in response to hypoglycemia is not defective if intraislet hyperinsulinemia is prevented. This suggests that exogenous insulin plays a pivotal role in the etiology of this defect.     

The influence of out-of-plane geometry on pulsatile flow within a distal end-to-side anastomosis

We present an experimental and computational investigation of time-varying flow in an idealized fully occluded 45 degrees distal end-to-side anastomosis. Two geometric configurations are assessed, one where the centerlines of host and bypass vessels lie within a plane, and one where the bypass vessel is deformed out of the plane of symmetry, respectively, termed planar and non-planar. Flow experiments were conducted by magnetic resonance imaging in rigid wall models and computations were performed using a high order spectral/hp algorithm. Results indicate a significant change in the spatial distribution of wall shear stress and a reduction of the time-averaged peak wall shear stress magnitude by 10% in the non-planar model as compared to the planar configuration. In the planar geometry the stagnation point follows a straight-line path along the host artery bed with a path length of 0.8 diameters. By contrast in the non-planar case the stagnation point oscillates about a center that is located off the symmetry plane intersection with the host artery bed wall, and follows a parabolic path with a 0.7 diameter longitudinal and 0.5 diameter transverse excursion. A definition of the oscillatory shear index (OSI) is introduced that varies between 0 and 0.5 and that accounts for a continuous range of wall shear stress vector angles. In both models, regions of elevated oscillatory shear were spatially associated with regions of separated or oscillating stagnation point flow. The mean oscillatory shear magnitude (considering sites where OSI>0.1) in the non-planar geometry was reduced by 22% as compared to the planar configuration. These changes in the dynamic behavior of the stagnation point and the oscillatory shear distribution introduced by out-of-plane graft curvature may influence the localization of vessel wall sites exposed to physiologically unfavorable flow conditions.     

Ca(2+)-induced movement of tropomyosin in skeletal muscle thin filaments observed by multi-site FRET

To obtain information on Ca(2+)-induced tropomyosin (Tm) movement in Ca(2+)-regulated muscle thin filaments, frequency-domain fluorescence energy transfer data were collected between 5-(2-iodoacetyl-amino-ethyl-amino)naphthalene-1-sulfonic acid at Cys-190 of Tm and phalloidin-tetramethylrhodamine B isothiocyanate bound to F-actin. Two models were used to fit the experimental data: an atomic coordinate (AC) model coupled with a search algorithm that varies the position and orientation of Tm on F-actin, and a double Gaussian distance distribution (DD) model. The AC model showed that little or no change in transfer efficiency is to be expected between different sites on F-actin and Tm if Ca(2+) causes azimuthal movement of Tm of the magnitude suggested by structural data (C. Xu, R. Craig, L. Tobacman, R. Horowitz, and W. Lehman. 1999. Biophys. J. 77:985-992). However, Ca(2+) produced a small but significant change in our phase/modulation versus frequency data, showing that changes in lifetime decay can be detected even when a change of the steady-state transfer efficiency is very small. A change in Tm azimuthal position of 17 on the actin filament obtained with the AC model indicates that solution data are in reasonable agreement with EM image reconstruction data. In addition, the data indicate that Tm also appears to rotate about its axis, resulting in a rolling motion over the F-actin surface. The DD model showed that the distance from one of the two chains of Tm to F-actin was mainly affected, further verifying that Ca(2+) causes Tm to roll over the F-actin surface. The width of the distance distributions indicated that the position of Tm in absence and in presence of Ca(2+) is well defined with appreciable local flexibility.     

Unsteady near wall residence times and shear exposure in model distal arterial bypass grafts

Building on previous studies of unsteady flow within model distal bypass grafts we analyse the near wall residence times and shear exposure in a 45 degrees anastomosis under symmetrical and symmetry breaking geometric configurations. We define residence time as the minimum time for a particle to exit a spherical region and shear exposure as a temporal integral of the Huber-Henky-von-Mises criterion along a particle path over a fixed time interval. Decomposing the pulsatile cycle into four equal intervals we find that the interval of peak residence time in the host vessel is from mid-deceleration to peak diastole and peak diastole to mid-acceleration. The asymmetric model is shown to have a significantly lower residence time during these intervals. Considering the shear exposure prior to the residence time evaluation we determine that a higher average shear exposure exists in the asymmetric model associated with the upstream geometry modification. Analysis of the regions of high residence time and shear exposure suggests that the "toe" region and the interface between the "heel" and bulk flow are more significant than the bed and heel region. Although the asymmetric model considered in this study reduces residence times in the host artery, the product of the measure of shear exposure and residence time is not found to be preferable. If shear exposure were to be considered as an important factor in particle activation, the findings imply that for junction optimisation, greater consideration needs to be given both to the local junction asymmetry and upstream influence on the shear history.     

Genetic Variation in Captive Koalas (Phascolarctos cinereus): Parentage Determination and Individual Identification

Highly repeatable randomly amplified polymorphicDNA (RAPD) markers were developed for parentage studiesin the koala (Phascolarctos cinereus). Of the 25 RAPDprimers screened, 5 (20.0%) produced 32 repeatable polymorphic RAPD bands (average/primer = 6.4± 4.2). A high level of polymorphism was observedfor each group of koalas (Featherdale, 71.9%; Lone Pine,84.4%). All 25 koalas could be uniquely identified using either RAPD or microsatellite markers. Of the32 RAPD markers generated in koalas, 25 were informativefor parentage analyses. These RAPD markers successfullydetermined both parents to three offspring and a male parent to a fourth offspring.Paternity analysis (where the female parent is known)succeeded in assigning the correct male parent to sevenoffspring. Our RAPD–PCR method generatesinformative genetic markers that are useful for parentagedetermination and individual identification of captivekoalas. This would provide genetic analysis to zoos andwildlife parks as a low-cost alternative to the more expensive microsatellite markers.     

Vestigial pit membrane remnants in perforation plates and helical thickenings in vessels of Ericaceae

Perforation plates from 15 species of 10 genera with scalarifom perforation plates, representing three subfamilies of woody Ericaceae (Rhododendroideae, Arbutoideae, Vaccinioideae) were studied with scanning electron microscopy (SEM). In most of them, pit membrane remnants were present, but these remnants were less extensive than in the ericalean families Clethraceae, Cyrillaceae, and Sarraceniaceae. Pit membrane remnants in perforations of vessels of Ericaceae are characteristically found at lateral ends of the perforations and in perforations (which may alternatively be called pits) transitional to lateral wall pitting. Pit membrane remnants were most extensive in Enkianthus. Phylogenetic and physiological factors for vestigial membrane presence in the perforations are discussed. Helical thickenings on vessel walls as seen with SEM are figured and described for Leucothoe and Pieris, and their significance is assessed.     

Ultrastructural observations on the erythrocytes and thrombocytes of the tuatara, Sphenodon punctatus (gray)

The erythrocytes of Sphenodon punctatus (Gray) are nucleated, ellipsoidal and flattened, and contain 55--65 microtubules in their marginal band. The thrombocytes are also flattened, ellipsoidal, nucleated cells and in electron-microscopic preparations occurred in aggregrates. The thrombocytes appeared to be 'activated' and possessed many pseudopodia which were devoid of organelles. The latter were concentrated in the perinuclear region and were encircled by a ring of microtubules. The organelles included ribosomes, mitochondria, membrane--bound dense material and numerous actin-like microfilaments. Cytoplasmic vacuoles contained a moderately dense, filamentous material and/or spheroidal electron-dense inclusions, beta-glycogen particles were scattered in the general cytoplasm and were most concentrated in the pseudopodia. The erythrocytes and thrombocytes of S. punctatus are compared with those in other vertebrates.     

PIT MEMBRANE REMNANTS IN PERFORATION PLATES OF PRIMITIVE DICOTYLEDONS AND THEIR SIGNIFICANCE

Perforations of vessel elements characteristically retain remnants of pit membranes (primary walls) in woods of species of more than 30 families of dicotyledons. Scanning electron microscopy is necessary to demonstrate presence and type of membrane remnant. Species with these remnants in perforations given in earlier literature as well as those newly reported here are listed. Perforation membrane remnants may take the form of flakes, strands, or webs, and particular types may characterize particular families (e.g., strands or bands in Illiciaceae). Some families have abundant perforation membrane remnants (e.g., Chloranthaceae, Illiciaceae). Where membranes are nearly intact, they are porose and closely resemble the porose pit membranes on end walls of Tetracentron tracheids. In Tetracentron, however, tracheary elements are monomorphic, so vessel origin cannot yet be said to have occurred. Membrane remnants in perforations are regarded as a relictual primitive feature that should be added to the list of primitive character states claimed for vessel elements in angiosperms; alternative hypotheses are considered and discussed, and evidence from DNA phylogenies is needed. In vessel-bearing dicotyledons with membrane remnants in perforations, many perforations are relatively clear, but an appreciable proportion of perforation plates do have membrane remnants.