ATP-binding on fibroblast growth factor 2 partially overlaps with the heparin-binding domain

Fibroblast growth factor 2 (FGF2), an intensively studied heparin-binding cytokine, is an important modulator of cell growth and differentiation under both physiological and pathophysiological conditions. It has been shown recently that ATP binds to FGF2 and that this binding is crucial for its biological function. In this study we demonstrated that divalent cations were not necessary for binding of ATP to FGF2, but it could be demonstrated that heparin blocked the labelling of FGF2 with ATP indicating an involvement of the heparin-binding domain (aa 128-144) in ATP-binding. FGF2, bound to Heparin Sepharose, could be eluted with ATP and GTP, but not with cAMP, AMP or ADP. Successive mutation of positively charged amino acid residues located in the heparin-binding domain drastically reduced the signal intensity of [gamma-(32)P]ATP labelled FGF2 indicating that this domain is not only important for heparin binding to FGF2 but also for ATP-binding.     

Cholesteryl ester hydroperoxides are biologically active components of minimally oxidized low density lipoprotein

Oxidation of low density lipoprotein (LDL) occurs in vivo and significantly contributes to the development of atherosclerosis. An important mechanism of LDL oxidation in vivo is its modification with 12/15-lipoxygenase (LO). We have developed a model of minimally oxidized LDL (mmLDL) in which native LDL is modified by cells expressing 12/15LO. This mmLDL activates macrophages inducing membrane ruffling and cell spreading, activation of ERK1/2 and Akt signaling, and secretion of proinflammatory cytokines. In this study, we found that many of the biological activities of mmLDL were associated with cholesteryl ester (CE) hydroperoxides and were diminished by ebselen, a reducing agent. Liquid chromatography coupled with mass spectroscopy demonstrated the presence of many mono- and polyoxygenated CE species in mmLDL but not in native LDL. Nonpolar lipid extracts of mmLDL activated macrophages, although to a lesser degree than intact mmLDL. The macrophage responses were also induced by LDL directly modified with immobilized 12/15LO, and the nonpolar lipids extracted from 12/15LO-modified LDL contained a similar set of oxidized CE. Cholesteryl arachidonate modified with 12/15LO also activated macrophages and contained a similar collection of oxidized CE molecules. Remarkably, many of these oxidized CE were found in the extracts of atherosclerotic lesions isolated from hyperlipidemic apoE(-/-) mice. These results suggest that CE hydroperoxides constitute a class of biologically active components of mmLDL that may be relevant to proinflammatory activation of macrophages in atherosclerotic lesions.     

Structural characterization of Ca(2+)-ATPase-bound phospholamban in lipid bilayers by solid-state nuclear magnetic resonance (NMR) spectroscopy

Phospholamban (PLN) regulates cardiac contractility by modulation of sarco(endo)plasmic reticulum calcium ATPase (SERCA) activity. While PLN and SERCA1a, an isoform from skeletal muscle, have been structurally characterized in great detail, direct information about the conformation of PLN in complex with SERCA has been limited. We used solid-state NMR (ssNMR) spectroscopy to deduce structural properties of both the A 36F 41A 46 mutant (AFA-PLN) and wild-type PLN (WT-PLN) when bound to SERCA1a after reconstitution in a functional lipid bilayer environment. Chemical-shift assignments in all domains of AFA-PLN provide direct evidence for the presence of two terminal alpha helices connected by a linker region of reduced structural order that differs from previous findings on free PLN. ssNMR experiments on WT-PLN show no significant difference in binding compared to AFA-PLN and do not support the coexistence of a significantly populated dynamic state of PLN after formation of the PLN/SERCA complex. A combination of our spectroscopic data with biophysical and biochemical data using flexible protein-protein docking simulations provides a structural basis for understanding the interaction between PLN and SERCA1a.     

Post-Fire Population Dynamics of Two Tree Species in High-Altitude Polylepis Forests of Central Ecuador

Fire is considered the main cause for the patchy distribution of high-montane tropical forests growing below the upper limit of tree growth, but there are little quantitative data on the impacts of burning on the respective tree populations. This study compares adult tree survival as well as sapling (0.05–1.3 m) and seedling (<0.05 m) recruitment of Polylepis incana , and the coexisting Gynoxis acostae in burned and unburned forest stands in the Páramo de Guamaní, central Ecuador. In P. incana , adult survival after burning was low, whereas all G. acostae individuals survived through resprouting. Two years after fire, the density of P. incana seedlings and saplings was higher than that of G. acostae , but still not sufficient for forest recovery. A sowing experiment revealed a significantly lower seedling emergence of both species in the burned than in the unburned plots. Seedling emergence was comparable to laboratory studies performed under optimal conditions, suggesting there was no evidence for climate constraining emergence at the given altitude. Interactions between seedling survival and burning for P. incana indicate higher seedling survival after burning, which could not be shown for G. acostae . Our data imply that single fire events strongly decrease adult and seedling population sizes in P. incana and thus may be the main reason for the discontinuous forest distribution below the upper distribution limit of the species. In contrast, the high resprouting potential of G. acostae explains its relatively high percentage in the remaining Ecuadorian P. incana stands.     

Lis1/dynactin regulates metaphase spindle orientation in Drosophila neuroblasts

Mitotic spindle orientation in polarized cells determines whether they divide symmetrically or asymmetrically. Moreover, regulated spindle orientation may be important for embryonic development, stem cell biology, and tumor growth. Drosophila neuroblasts align their spindle along an apical/basal cortical polarity axis to self-renew an apical neuroblast and generate a basal differentiating cell. It is unknown whether spindle alignment requires both apical and basal cues, nor have molecular motors been identified that regulate spindle movement. Using live imaging of neuroblasts within intact larval brains, we detect independent movement of both apical and basal spindle poles, suggesting that forces act on both poles. We show that reducing astral microtubules decreases the frequency of spindle movement, but not its maximum velocity, suggesting that one or few microtubules can move the spindle. Mutants in the Lis1/dynactin complex strongly decrease maximum and average spindle velocity, consistent with this motor complex mediating spindle/cortex forces. Loss of either astral microtubules or Lis1/dynactin leads to spindle/cortical polarity alignment defects at metaphase, but these are rescued by telophase. We propose that an early Lis1/dynactin-dependent pathway and a late Lis1/dynactin-independent pathway regulate neuroblast spindle orientation.     

Influence of canopy aspect and height on codling moth (Lepidoptera: Tortricidae) larval infestation in apple, and relationship between infestation and fruit size

Monitoring systems based on traps with female attractants are expected to enhance forecasting of insect population size and damage. The optimal placement of such traps should match the small-scale distribution of ovipositing females. In the codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), fruit infestation takes place in proximity to the oviposition site. We characterized the within-tree distribution of codling moth infestations and the size of uninfested fruit based on a survey of 40,000 apples (Malus spp.) from trees belonging to 160 different apple genotypes and growing in two different environments. Each tree was subdivided into 12 sectors, considering canopy aspect (north, east, south, and west) and canopy height (bottom, middle, and top). This study revealed that fruit infestation by the first but not by the second generation of larvae correlated significantly with canopy aspect. Similarly, fruit size differed significantly between the north- and the south-facing tree side for the period of infestation by the first but not by the second larval generation. Significantly lower fruit infestation was observed on the north- compared with the south- or east-facing tree side for the first generation. A significant influence of canopy height on larval infestation was observed in three of eight assessments, in which the middle height level showed the highest infestations. Significant differences in within-tree distribution of codling moth infestation suggest that oviposition preference is guided by nonrandom factors including microclimate, fruit phenology, and wind direction. These cultivar-independent findings should be considered in future monitoring systems that focus on female codling moth.     

Postnatal life events affect the severity of asthmatic airway inflammation in the adult rat

Genetic and hygienic factors influence susceptibility to asthma. In autoimmune and inflammatory diseases, additional effects of the psychosocial environment have been demonstrated that might also play a role in asthma. In this study, the impact of different early postnatal stressors on an OVA-induced model of asthma was tested in adulthood. Fischer 344 rats were subjected to either repeated handling stimulation (HA), maternal separation (MS), or were left undisturbed in their first 4 wk of life. Behavioral differences were characterized at the age of 4 mo. At 5 mo of age, immunological cellular and serologic changes were investigated and experimental asthma was induced. Results show significantly increased exploratory behavior and reduced anxiety in HA rats compared with MS and controls. Without further behavioral or immunological challenges, HA animals exhibited an increased ex vivo NK cell cytotoxicity but no other obvious immunological differences. After induction of asthma, in contrast, MS animals exhibited proinflammatory effects in leukocyte subset composition including increased eosinophil numbers, whereas levels of IgE and the allergy-specific cytokine IL-13 were reduced compared with HA. There was a most remarkable increase of adrenocorticotropin in HA animals, comparing pre- to postchallenge plasma levels. These data demonstrate for the first time that early postnatal stimulative or adverse experiences exert long-lasting changes of the "neuroendocrinoimmune" interface in adulthood, resulting in either protective or aggravating mechanisms in allergic airway disease. Thus, in addition to genetic and hygienic factors, nongenetically acquired individual differences contribute to the pathobiology of asthma.     

Responses of arthropods to plant diversity: changes after pollution cessation

Data collected from three different polluted sites in the vicinity of a phosphate fertilizer factory that was closed in 1990 are used to test with Mantel tests and smoothing techniques whether the rapid increase of plant species richness following cessation of pollution enhanced associated arthropod assemblage diversity. 132 plant species (between 1990 and 1999) and 66 413 individuals of 680 arthropod species (using sweep net sampling between 1990 and 1996) were recorded. Using top soil pH as a representative pollution parameter we detected an increase of plant species richness, effective diversity and evenness of plant community with decreasing pH both in space and time. While the richness of all studied functional groups of herbivores increased with plant species richness, only the richness of one carnivore functional group showed a similar pattern. Plant species richness was significantly correlated to the abundance patterns of two herbivore and two carnivore groups. But contrary to theoretical predictions consumer abundance tended to decrease with increasing plant diversity only between a plant species richness range of 10 to ca 35. Our results support the findings of previous studies that highlight how increased plant species and functional group richness may result in higher herbivore species richness, and that carnivore richness may be influenced by herbivore and detritivore richness. The functional group approach used in this study has enabled us to detected the very individual interaction patterns that occur between different groups within the same trophic level.     

Marine metapopulations: a useful concept?

We discuss the potential and limitations of the metapopulation concept in marine ecology. The usefulness of the concept in terrestrial ecology is neither based on its simplicity or generality nor on overwhelming empirical evidence. The usefulness is in the questions which are asked when the metapopulation concept is applied. These questions address spatial phenomena and processes on different spatial scales. They help in acknowledging that every population, be it terrestrial or marine, has a spatial organization. Understanding this spatial organization is also important for tackling specific applied problems, i.e. to avoid overexploitation of living marine resources or for configuring marine reserves. The 'openness' of coastal populations, whose larvae enter larval pools or which are holoplanktonic, is no reason for not asking the questions implied by the metapopulation concept. For marine ecology, the real problem is to delineate populations, which then may possibly correspond to the 'local populations' of metapopulations. Thus, the answer to the question in the title of this paper, whether 'marine metapopulation' is a useful concept, is 'yes', if the concept is considered a working hypotheses, if the concept is explicitly defined, and if the questions linked to the concept are clearly stated. Even if it eventually transpires that only very few marine metapopulations actually exist, marine ecology would still have gained some important new insights. Electronic Publication     

Empty habitat in coastal sediments for populations of macrozoobenthos

Species with wide dispersal are expected to have little empty habitat. This was tested by analysing habitat use in the Wadden Sea near the island of Sylt (North Sea). Sampling covered 222 intertidal sites spread across the K?nigshafen tidal flats (4.5 km², mapping approach conducted once) and 270 subtidal sites along 12 km of a tidal channel system (stratified random sampling in spring and autumn). For any single species, 'suitable habitat' was extrapolated from the ranges of water depth and sediment composition present at the ten sites with the highest frequency or abundance of the species. On average, macrobenthic species actually used less than half of the suitable sites within the scale of local populations; this was far less than expected from a lognormal distribution. In the subtidal, abundance of most species changed between the two sampling seasons. A 50% increase in overall abundance was accompanied by a decrease in empty habitat of only about 25%. Thus, a doubling of abundances would not fill the empty space but just of half of it. The polychaete Scoloplos armiger was an exception in occupying almost all of its suitable habitat in the intertidal and subtidal sediments. A distinct patch of high species richness occurred where flood waters persistently form a large gyre which may enhance larval settlement. We suggest that limitations to larval settlement and/or juvenile survival primarily cause the extent of the observed habitat emptiness. Electronic Publication