RAPID ALDOSTERONE-INDUCED CELL VOLUME INCREASE OF ENDOTHELIAL CELLS MEASURED BY THE ATOMIC FORCE MICROSCOPE

Atomic force microscopy (AFM) is a useful technique for imaging the surface of living cells in three dimensions. The authors applied AFM to obtain morphological information of individual cultured endothelial cells of bovine aorta under stationary and strain conditions and to simultaneously measure changes in cell volume in response to aldosterone. This mineralocorticoid hormone is known to have acute, non-genomic effects on intracellular pH, intracellular electrolytes and inositol-1,4,5-triphosphate production. In this study whether endothelial cells under tension change their volume in response to aldosterone was tested. Such changes were already shown in human leukocytes measured by Coulter counter. In contrast to leukocytes that are more or less spherical and live in suspension, endothelial cells exhibit a complex morphology and adhere to a substrate. Thus, measurements of discrete cell volume changes in endothelial cells under physiological condition is only feasible with more sophisticated techniques. By using AFM we could precisely measure the absolute cell volume of individual living endothelial cells. Before the addition of aldosterone the cell volume of mechanically stressed endothelial cells mimicking arterial blood pressure was 1827±172fl. Cell volume was found to increase by 28% 5min after hormone exposure. Twenty-five minutes later cell volume was back to normal despite the continuous presence of aldosterone in the medium. Amiloride, a blocker of the plasma membrane Na⁺/H⁺exchanger prevented the initial aldosterone-induced volume increase. Taken together, AFM disclosed a transient swelling of endothelial cells induced by the activation of an aldosterone sensitive plasma membrane Na⁺/H⁺exchanger.     

Apioceridae (Insecta: Diptera): cladistic reappraisal and biogeography

Twenty members of the fly families Apioceridae and Mydidae are compared in terms of 77 adult characters. Cladistic analysis of 91 synapomorphies provides a well corroborated reconstruction of lineage relationships, and reveals that the Apioceridae is paraphyletic with respect to the Mydidae. In order to render the Apioceridae monophyletic, the genus Rhaphiomidas and the subfamily Megascelinae of the Apioceridae are transferred to the Mydidae. The subfamily Rhaphiomidinae is reinstated to accommodate Rhaphiomidas , comprising the most plesiomorphic mydids. The relationships of the remaining subfamilies of Mydidae are discussed in the context of these findings. The genus Apiocera is divided into four subgenera, the type subgenus Apiocera Westwood in Australia, Pyrocera subgen. nov. in North America, and the subgenera Ripidosyrma Hermann and Anypenus Philippi are applied to the South African and South American species, respectively. A key to the four subgenera of Apiocera is provided. The biogeographic relationships of the subgenera and genera of the Apioceridae and the Megascelinae are discussed. Although considered an example of a transantarctic or Gondwanan group, we argue that the distribution of the Apioceridae predates the Mesozoic supercontinent Gondwanaland and extends onto sections of Pangaea, and should be termed 'Pangaean'. The cladistic relationships between the genera of Apioceridae and Megascelinae are consistent with the geological vicariance of the fragments of Pangaea on which they now occur.     

A TOURNAISIAN BRACHIOPOD FAUNA FROM SOUTH-EAST WALES

Abstract:  An exceptionally preserved fauna within dolomites of the Friars Point Limestone Formation includes the most diverse brachiopod assemblage yet described from the Tournaisian of the British Isles, and the first from Wales. Each of the 16 brachiopod genera includes a single species, of which four are new ( Schellwienella cheuma , Schuchertella subcrona , Composita ptygmation , Fusella extrata ). Associated fossils are corals (one species), bryozoans (two species) and crinoids (one species). Spiriferoideans and schizophorides are numerically dominant, indicative of level-bottom, inner mid-ramp biotopes. Biogeographical comparisons reflect the cosmopolitanism of early Carboniferous brachiopod generic assemblages. Taxonomic comparisons involve selection of lectotypes for Syringothyris exoleta North, 1920, Syringothyris cyrtorhyncha North, 1920, Tylothyris laminosa beta North, 1920, and Tylothyris laminosa gamma North, 1920.     

Seasonal changes of osmotic pressure, symplasmic water content and tissue elasticity in the blades of dune grasses growing in situ along the coast of Oregon

Abstract Midday water potentials of blades of the dune grasses Ammophila arenaria (L.) Link and Elymus mollis Trin. ex Spreng. growing in situ declined over the summer growing period, indicating a trend of increasing water stress. An analysis of the water relations characteristics of these blades using pressure-volume techniques demonstrated that both species increased bulk osmotic pressure at full hydration () and, therefore, bulk turgor as an acclimation response. In A. arenaria, however, the increase of osmotic pressure (+ 0.35 MPa) was entirely the result of decreasing symplasmic water content. The increase of osmotic pressure (+ 0.54 MPa) observed in E. mollis blades was due to solute accumulation (72% of Δ) and to a lesser degree, decreased symplasmic water content (28% of Δ). Osmotic adjustment in E. mollis blades was accompanied by a significant decrease in tissue elasticity (_max went from 12 to 19 MPa). The elastic properties of A. arenaria blades remained constant over the same period and had a maximum modulus (10 MPa) that was always less than that of E. mollis, As estimated from Höfler plots, these seasonal adjustments of osmotic pressure and differences in tissue elasticity enabled plants in situ to maintain turgor pressure in the range of 0.5–0.6 MPa at the lowest water potentials of mid-August. Laboratorygrown plants exhibited the species-specific differences in osmotic pressure, turgor pressure, and tissue elasticity observed in field plants. Although certain alterations of leaf structure were expected to coincide with the observed changes and species-specific differences in symplasmic water content and tissue elasticity, these could not be detected by measurements of specific leaf weight or the ratio of dry matter to saturated water content.     

Genetic differentiation amongst fragrant orchids (Gymnadenia conopsea s.l.) in the British Isles

Genetic variation was examined in five microsatellite loci to seek evidence of genetic differentiation and restricted gene flow that would support the taxonomic division of Gymnadenia into three species ( G. borealis , G. conopsea , and G. densiflora ). A total of 107 alleles was detected in 17 populations from England, Scotland, and Ireland. The mean expected heterozygosities ranged from 0.48 to 0.81. The differentiation in allele frequencies amongst populations that had been assigned to each taxon on the basis of morphology was sufficiently large to support the taxa as distinct species. Phylogenetic trees based on microsatellite allele frequencies, as well as assignment tests, supported the existence of three distinct groups with at least partial restriction of gene flow between them. There was substantial homozygote excess, leading to high F _IS estimates, for most loci in most populations. This is unlikely to have been a result of widespread null alleles, and more probably reflects a high level of inbreeding in G. conopsea . This inference requires further investigation. The implications of the results of this and other taxonomic studies for the conservation of Gymnadenia in Britain are discussed.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 349–360.     

Are Roots a Source of Abscisic Acid for the Shoots of Flooded Pea Plants?

Flooding the soil for 2–5 d decreased stomatal conductancesof pea plants (Pisum sativum L., cv. Sprite) with six or sevenleaves. This coincided with slower transpiration, increasedleaf water potentials and increased concentrations of abscisicacid (ABA) in the leaves. No increase in ABA was found in theterminal 20 mm of roots of flooded plants over the same timeperiod. Small stomatal conductances associated with increases in foliarABA were also found in plants grown in nutrient solution whenaeration was halted, causing the equilibrium partial pressuresof dissolved oxygen to fall below 05 It Pa. No increase in ABAconcentration in young secondary roots of the non-aerated plantswas detected after 24, 48 or 72 h, even when the shoot, thepresumed site of deposition for any ABA from the roots, wasremoved 5–6 h before analysis. Similarly, ABA concentrations in roots were not increased whenthe nutrient solution was de-oxygenated by continuous purgingwith nitrogen gas. The abscisic acid concentration in leaf epidermis,the tissue most likely to be the recipient of any ABA movingin the transpiration stream from oxygen-deficient roots, waslower than in the remaining parts of the leaf when examinedin the mutant Argenteum which possesses easily removable epidermallayers. It is concluded that the leaves of plants subjectedto flooding of the soil or oxygen shortage in the root environmentare not enriched substantially with ABA from the roots. A moreprobable source of this growth regulator is the leaf itself. Key words: Pisum sativum, flooding, roots, hormones, aeration stress, abscisic acid, Argenteum mutant     

Reconciling the optimal and empirical approaches to modelling stomatal conductance

Models of vegetation function are widely used to predict the effects of climate change on carbon, water and nutrient cycles of terrestrial ecosystems, and their feedbacks to climate. Stomatal conductance, the process that governs plant water use and carbon uptake, is fundamental to such models. In this paper, we reconcile two long‐standing theories of stomatal conductance. The empirical approach, which is most commonly used in vegetation models, is phenomenological, based on experimental observations of stomatal behaviour in response to environmental conditions. The optimal approach is based on the theoretical argument that stomata should act to minimize the amount of water used per unit carbon gained. We reconcile these two approaches by showing that the theory of optimal stomatal conductance can be used to derive a model of stomatal conductance that is closely analogous to the empirical models. Consequently, we obtain a unified stomatal model which has a similar form to existing empirical models, but which now provides a theoretical interpretation for model parameter values. The key model parameter, g₁, is predicted to increase with growth temperature and with the marginal water cost of carbon gain. The new model is fitted to a range of datasets ranging from tropical to boreal trees. The parameter g₁ is shown to vary with growth temperature, as predicted, and also with plant functional type. The model is shown to correctly capture responses of stomatal conductance to changing atmospheric CO₂, and thus can be used to test for stomatal acclimation to elevated CO₂. The reconciliation of the optimal and empirical approaches to modelling stomatal conductance is important for global change biology because it provides a simple theoretical framework for analyzing, and simulating, the coupling between carbon and water cycles under environmental change.     

Microsatellite DNA markers for the study of Atlantic salmon (Salmo salar) kinship,population structure,and mixed‐fishery analyses

Eleven microsatellite DNA loci were identified and characterized for Atlantic salmon (Salmo salar) collected from the Penobscot River, Maine, USA and the River Nith, Scotland, UK. The markers revealed high levels of genetic diversity (seven to 48 alleles per locus), heterozygosity (to 100%), and allelic heterogeneity (all comparisons). Considerable differentiation was observed as the genetic distance (chord) between the two collections was 0.680 and the pairwise F_ST, 0.12, was highly significant. These findings are consistent with patterns of continental‐level differentiation observed previously using an alternate suite of microsatellite loci. Locus‐by‐locus analyses of molecular variance suggested that most markers were suitable for delineating kinships and population genetic structure.