Foliar and wood chemistry of sugar maple along a gradient of soil acidity and stand health

The decline of sugar maple (Acer saccharum Marsh.) in forest of north-eastern North America is an important environmental issue. In this study, relationships between, soil, wood and foliar chemistry were assessed for 17 stands distributed within a large area of the Quebec sugar maple forest and that were growing on soils with a strong gradient of acidity and base saturation. There were many significant relationships between variables describing the acid-base status of the top-B soil (Ca and Mg concentrations, exchangeable acidity and base saturation) and Ca and Mn concentrations and Ca/Mn and Mg/Mn in tree tissues. Manganese was the element that showed the strongest inverse non-linear relationships with top-B soil base saturation with variance explanation of 71 and 65%, for wood and foliage, respectively. The 17 sites were divided in two groups according to their level of decline. The declining stands had significantly higher wood Mn and Mg concentrations and lower Ca/Mn ratios and significantly higher foliar Mn and lower Ca and Al concentrations. It was impossible to determine if these differences were a cause or a symptom of sugar maple health. However, the increase in Mn concentrations in tree tissues with increasing soil acidity, as well as the higher Mn concentrations in declining as compared to healthy stands suggest that Mn, as well as low Ca availability, could be an important contributing factor in the sugar maple decline.     

Evaluation of the <Emphasis Type="Italic">OPTC</Emphasis> gene in primary open angle glaucoma: functional significance of a silent change

Background  

We investigated the molecular basis of primary open-angle glaucoma (POAG) using Opticin (OPTC) as a candidate gene on the basis of its expression in the trabecular meshwork cells involved in the disease pathogenesis. Two hundred POAG patients and 100 controls were enrolled in this study. The coding sequence of OPTC was amplified by PCR from genomic DNA of POAG patients, followed by SSCP, DHPLC and DNA sequencing. Subsequent bioinformatic analysis, site-directed mutagenesis, quantitative RT-PCR and western blot experiments were performed to address the functional significance of a 'silent' change in the OPTC coding region while screening for mutations in POAG patients.     

Mutation accumulation and the effect of copia insertions in Drosophila melanogaster

Repeated efforts to estimate the genomic deleterious mutation rate per generation (U) in Drosophila melanogaster have yielded inconsistent estimates ranging from 0.01 to nearly 1. We carried out a mutation-accumulation experiment with a cryopreserved control population in hopes of resolving some of the uncertainties raised by these estimates. Mutation accumulation (MA) was carried out by brother sister mating of 150 sublines derived from two inbred lines. Fitness was measured under conditions chosen to mimic the ancestral laboratory environment of these genotypes. We monitored the insertions of a transposable element, copia, that proved to accumulate at the unusually high rate of 0.24 per genome per generation in one of our MA lines. Mutational variance in fitness increased at a rate consistent with previous studies, yielding a mutational coefficient of variation greater than 3%. The performance of the cryopreserved control relative to the MA lines was inconsistent, so estimates of mutation rate by the Bateman-Mukai method are suspect. Taken at face value, these data suggest a modest decline in fitness of about 0.3% per generation. The element number of copia was a significant predictor of fitness within generations; on average, insertions caused a 0.76% loss in fitness, although the confidence limits on this estimate are wide.     

Glucocorticoid ligands specify different interactions with NF-kappaB by allosteric effects on the glucocorticoid receptor DNA binding domain

Glucocorticoids inhibit inflammation by acting through the glucocorticoid receptor (GR) and powerfully repressing NF-kappaB function. Ligand binding to the C-terminal of GR promotes the nuclear translocation of the receptor and binding to NF-kappaB through the GR DNA binding domain. We sought how ligand recognition influences the interaction between NF-kappaB and GR. Both dexamethasone (agonist) and RU486 (antagonist) promote efficient nuclear translocation, and we show occupancy of the same intranuclear compartment as NF-kappaB with both ligands. However, unlike dexamethasone, RU486 had negligible activity to inhibit NF-kappaB transactivation. This failure may stem from altered co-factor recruitment or altered interaction with NF-kappaB. Using both glutathione S-transferase pull-down and bioluminescence resonance energy transfer approaches, we identified a major glucocorticoid ligand effect on interaction between the GR and the p65 component of NF-kappaB, with RU486 inhibiting recruitment compared with dexamethasone. Using the bioluminescence resonance energy transfer assay, we found that RU486 efficiently recruited NCoR to the GR, unlike dexamethasone, which recruited SRC1. Therefore, RU486 promotes differential protein recruitment to both the C-terminal and DNA binding domain of the receptor. Importantly, using chromatin immunoprecipitation, we show that impaired interaction between GR and p65 with RU486 leads to reduced recruitment of the GR to the NF-kappaB-responsive region of the interleukin-8 promoter, again in contrast to dexamethasone that significantly increased GR binding. We demonstrate that ligand-induced conformation of the GR C-terminal has profound effects on the functional surface generated by the DNA binding domain of the GR. This has implications for understanding ligand-dependent interdomain communication.     

Regulation of vascular endothelial growth factor receptor 2-mediated phosphorylation of focal adhesion kinase by heat shock protein 90 and Src kinase activities

Exposure of endothelial cells to vascular endothelial growth factor (VEGF) induced tyrosine phosphorylation of focal adhesion kinase (FAK) on site Tyr(407), an effect that required the association of VEGF receptor 2 (VEGFR2) with HSP90. The association of VEGFR2 with HSP90 involved the last 130 amino acids of VEGFR2 and was blocked by geldanamycin, a specific inhibitor of HSP90. Moreover, geldanamycin inhibited the VEGF-induced activation of the small GTPase RhoA, which resulted in an inhibition of phosphorylation of FAK on site Tyr(407). In this context, the inhibition of RhoA kinase (ROCK) with Y27632 or by expression of dominant negative forms of RhoA or ROCK impaired the VEGF-induced phosphorylation of Tyr(407) within FAK. In contrast to phosphorylation of Tyr(861), the phosphorylation of site Tyr(407) was insensitive to Src kinase inhibition by 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d] pyrimidine (PP2). We also found that the recruitment of paxillin to FAK was inhibited by geldanamycin but not by PP2, whereas both geldanamycin and PP2 inhibited the recruitment of vinculin to FAK. In accordance, the recruitment of paxillin and vinculin to FAK was inhibited in cells that express the mutant FAK-Y407F, whereas the expression of the mutant Y861F inhibited the recruitment of paxillin but not of vinculin. Importantly, cell migration was abolished in cells in which the signal from the VEGFR2-HSP90 pathway was blocked by the expression of Delta130VEGFR2, a deletant of VEGFR2 that does not associate with HSP90. Our findings underscore for the first time the key role played by the VEGFR2-HSP90-RhoA-ROCK-FAK/Tyr(407) pathway in transducing the VEGF signal that leads to the assembly of focal adhesions and endothelial cell migration.     

Cdx1 autoregulation is governed by a novel Cdx1-LEF1 transcription complex

The Cdx1 gene product is essential for normal anterior-posterior vertebral patterning. Expression of Cdx1 is regulated by several pathways implicated in anterior-posterior patterning events, including retinoid and Wnt signaling. We have previously shown that retinoic acid plays a key role in early stages of Cdx1 expression at embryonic day 7.5 (E7.5), while both Wnt3a signaling and an autoregulatory loop, dependent on Cdx1 itself, are involved in later stages of expression (E8.5 to E9.5). This autoregulation is reflected by the ability of Cdx1 to affect expression from proximal Cdx1 promoter sequences in tissue culture. However, this region is devoid of a demonstrable Cdx response element(s). We have now found that Cdx1 and LEF1, a nuclear effector of Wnt signaling, synergize to induce expression from the Cdx1 promoter through previously documented LEF/T-cell factor response elements. We also found a direct physical interaction between the homeodomain of Cdx1 and the B box of LEF1, suggesting a basis for this synergy. Consistent with these observations, analysis of Cdx1 Wnt3a(vt) compound mutants demonstrated that Wnt and Cdx1 converged on Cdx1 expression and vertebral patterning in vivo. Further data suggest that Cdx-high-mobility group box interactions might be involved in a number of additional pathways.     

The N Terminus of the OB Domain of Telomere Protein TPP1 Is Critical for Telomerase Action

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Interactions between resources and abiotic conditions control plant performance on subarctic coastal dunes

Both abiotic conditions and resource levels affect the performance of plants on coastal dune systems. On the foredune, environmental factors are particularly limiting for plant growth and these vary along a short topographical gradient, from the foot to the ridge. On subarctic coastal dunes in northeastern Canada, this topographical gradient is paralleled by a plant sequence that typically involves Honckenya peploides, Elymus mollis, and Lathyrus japonicus. In this study, field nutrient additions were carried out to evaluate the importance of N and/or P limitation on foredune plant performance. Also, glasshouse experiments were done to determine the significance of interactions between substrate resources (i.e., nutrients and water), and between substrate resources (i.e., nutrients) and an abiotic condition (i.e., salt spray) on the growth of a dune species. Field nutrient additions did not result in any significant increase in plant biomass, although nutrients were accumulated in the rhizomes of all three species present on the foredune and in the aboveground tissues of Elymus. Glasshouse experiments on Elymus showed that nutrient addition could increase plant biomass. However, water availability and salt spray interfered with nutrient use by the plants. I suggest that such interactions between resources and abiotic conditions may significantly affect plant performance and plant sequence on the foredune of coastal dune systems.     

The influence of understory nectar resources on parasitism of the spruce budworm Choristoneura fumiferana in the field

1 Nectar provided by flowering plants has been suggested as an important factor enhancing parasitism rates of herbivorous insects. Artificial nectar has been shown to increase parasitoid longevity and fecundity in laboratory studies. 2 We studied the influence of understory nectar on parasitism of the spruce budworm Choristoneura fumiferana in a field experiment in which we either removed understory flowering herbs or sprayed a sucrose solution on the understory vegetation in 0.25 ha quadrats. 3 Cohorts of laboratory-reared second instars were transferred the field to evaluate parasitism in the quadrats. The larvae were harvested as fifth instars and reared for parasitoids. 4 Parasitoids of early instars caused greater mortality in the supplemental sucrose treatment than in the vegetation-removal treatment or the control. Parasitoids of older larvae did not respond to the treatments. Larger-scale treatments may be necessary to elicit a response by the larger parasitoids that attack older larvae.