Fibroblast growth factor receptor-1-mediated endothelial cell proliferation is dependent on the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk.

Stimulation of fibroblast growth factor receptor-1 (FGFR-1) expressed on endothelial cells leads to cellular migration and proliferation. We have examined the role of the Src homology (SH) 2/SH3 domain-containing adaptor protein Crk in these processes. Transient tyrosine phosphorylation of Crk in fibroblast growth factor-2-stimulated endothelial cells was dependent on the juxtamembrane tyrosine residue 463 in FGFR-1, and a Crk SH2 domain precipitated FGFR-1 via phosphorylated Tyr-463, indicating direct complex formation between Crk and FGFR-1. Furthermore, Crk SH2 and SH3 domains formed ligand-independent complexes with Shc, C3G, and the Crk-associated substrate (Cas). Tyrosine phosphorylation of C3G and Cas increased as a consequence of growth factor treatment. We examined the role of Crk in FGFR-1-mediated cellular responses by use of cells expressing chimeric platelet-derived growth factor receptor-alpha/FGFR-1 (alphaR/FR) wild type and mutant Y463F receptors. The kinase activity of alphaR/FR Y463F was intact, but both Crk and the adaptor FRS-2 were no longer tyrosine-phosphorylated in the mutant cells. Both wild type and mutant receptor cells migrated efficiently, whereas cells expressing the mutant alphaR/FR Y463F failed to proliferate and Erk2 and Jun kinase activities were suppressed in these cells. In wild type alphaR/FR cells transiently expressing an SH2 domain mutant of Crk, Erk and Jun kinase activities as well as DNA synthesis were attenuated. Our data indicate that Crk participates in signaling complexes downstream of FGFR-1, which propagate mitogenic signals.     

Different combinations of laccase paralogs nonredundantly control the amount and composition of lignin in specific cell types and cell wall layers in Arabidopsis

Vascular plants reinforce the cell walls of the different xylem cell types with lignin phenolic polymers. Distinct lignin chemistries differ between each cell wall layer and each cell type to support their specific functions. Yet the mechanisms controlling the tight spatial localization of specific lignin chemistries remain unclear. Current hypotheses focus on control by monomer biosynthesis and/or export, while cell wall polymerization is viewed as random and nonlimiting. Here, we show that combinations of multiple individual laccases (LACs) are nonredundantly and specifically required to set the lignin chemistry in different cell types and their distinct cell wall layers. We dissected the roles of Arabidopsis thaliana LAC4, 5, 10, 12, and 17 by generating quadruple and quintuple loss-of-function mutants. Loss of these LACs in different combinations led to specific changes in lignin chemistry affecting both residue ring structures and/or aliphatic tails in specific cell types and cell wall layers. Moreover, we showed that LAC-mediated lignification has distinct functions in specific cell types, waterproofing fibers, and strengthening vessels. Altogether, we propose that the spatial control of lignin chemistry depends on different combinations of LACs with nonredundant activities immobilized in specific cell types and cell wall layers.

The spatial control of lignin chemistry, and thus of specific cellular functions, depends on combinations of laccases with nonredundant activities in specific cell types and cell wall layers.

IN A NUTSHELL Background: Lignins are a diverse, complex group of aromatic polymers that accumulate in cell walls of vascular plants, reinforcing organs, and enabling long-distance water transport. The different cell wall layers of each cell type exhibit specific lignin chemistries with distinct proportions of specific aromatic substitutions and aliphatic functions. The spatial control of this lignin chemistry was supposed to depend exclusively on the chemical identity of the lignin monomers exported into the cell wall. However, monomer supply alone cannot fully explain the sharp spatial differences between each cell wall layer in the different cell types. We, therefore, investigated whether different paralogs of the lignin monomer-oxidizing LACCASE enzymes are responsible for spatially controlling lignin chemistry at the cell wall layer level for the different cell types in the vascular tissues of plants. Question: How are specific lignin chemistries spatially controlled by LACCASE paralogs in each cell wall layer and cell type? What are the roles of LACCASE-dependent lignin accumulation for the mechanical reinforcement and the waterproofing of different cell types in plant vascular tissues? Findings: We answered these questions by identifying the LACCASE paralogs specifically expressed in vascular cells undergoing lignin accumulation. We analyzed their functions using genetic engineering to switch off five of the six LACCASE paralog genes associated with lignin formation. Their importance in the cell wall layer and cell type lignin accumulation was determined by comparing plants sharing four of the five mutations in different LACCASE paralogs. We show that each LACCASE paralog exhibits specific substrate preference, pH optimum and localization differing between the cell wall layers of each cell type. Their lignin concentration and composition moreover depended on specific combinations of LACCASE paralogs, each enabling different aromatic substitutions and aliphatic functions to accumulate. Impairing these LACCASE-dependent lignin chemistries resulted in the loss of cell wall mechanical resistance of sap-conducting cells and the loss of cell wall waterproofing of organ-reinforcing fiber cells. Next steps: We are now pursuing research to understand the molecular mechanisms controlling the supply of lignin precursors as well as the temporal regulation activating lignification during the formation/maturation of each cell wall layer in the different cell types.     

Is hunting mortality additive or compensatory to natural mortality? Effects of experimental harvest on the survival and cause-specific mortality of willow ptarmigan

1. The effects of harvest on the annual and seasonal survival of willow ptarmigan Lagopus lagopus L. were tested in a large-scale harvest experiment. Management units were randomly assigned to one of three experimental treatments: 0%, 15% or 30% harvest. Seasonal quotas were based on the experimental treatment and estimates of bird density before the hunting season. Survival rates and hazard functions for radio-marked ptarmigan were then estimated under the competing risks of harvest and natural mortality. 2. The partially compensatory mortality hypothesis was supported: annual survival of ptarmigan was 0·54 ± 0·08 SE under 0% harvest, 0·47 ± 0·06 under 15% harvest, and was reduced to 0·30 ± 0·05 under 30% harvest. Harvest mortality increased linearly from 0·08 ± 0·05, 0·27 ± 0·05 and 0·42 ± 0·06 from 0% to 30% harvest, whereas natural mortality was 0·38 ± 0·08, 0·25 ± 0·05 and 0·28 ± 0·06 under the same treatments. 3. Realized risk of harvest mortality was 0·08-0·12 points higher than our set harvest treatments of 0-30% because birds were exposed to risk if they moved out of protected areas. The superadditive hypothesis was supported because birds in the 30% harvest treatment had higher natural mortality during winter after the hunting season. 4. Natural mortality was mainly because of raptor predation, with two seasonal peaks in fall and spring. Natural and harvest mortality coincided during early autumn with little potential for compensation during winter months. Peak risk of harvest mortality was 5× higher than natural mortality. Low natural mortality during winter suggests that most late season harvest would be additive mortality. 5. Environmental correlates of natural mortality of ptarmigan included seasonal changes in snow cover, onset of juvenile dispersal, and periods of territorial activity. Natural mortality of ptarmigan was highest during autumn movements and nesting by gyrfalcons Falco rusticolus L. Mortality was low when gyrfalcons had departed for coastal wintering sites, and during summer when ptarmigan were attending nests and broods. 6. Our experimental results have important implications for harvest management of upland gamebirds. Seasonal quotas based on proportional harvest were effective and should be set at ≤ 15% of August populations for regional management plans. Under threshold harvest of a reproductive surplus, 15% harvest would be sustainable at productivity rates ≥ 2·5 young per pair. Impacts of winter harvest could be minimized by closing the hunting season in early November or by reducing late season quotas.     

Breast cancer risk and 6q22.33: combined results from Breast Cancer Association Consortium and Consortium of Investigators on Modifiers of BRCA1/2

Recently, a locus on chromosome 6q22.33 (rs2180341) was reported to be associated with increased breast cancer risk in the Ashkenazi Jewish (AJ) population, and this association was also observed in populations of non-AJ European ancestry. In the present study, we performed a large replication analysis of rs2180341 using data from 31,428 invasive breast cancer cases and 34,700 controls collected from 25 studies in the Breast Cancer Association Consortium (BCAC). In addition, we evaluated whether rs2180341 modifies breast cancer risk in 3,361 BRCA1 and 2,020 BRCA2 carriers from 11 centers in the Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA). Based on the BCAC data from women of European ancestry, we found evidence for a weak association with breast cancer risk for rs2180341 (per-allele odds ratio (OR)?=?1.03, 95% CI 1.00-1.06, p?=?0.023). There was evidence for heterogeneity in the ORs among studies (I(2)?=?49.3%; p?=?<0.004). In CIMBA, we observed an inverse association with the minor allele of rs2180341 and breast cancer risk in BRCA1 mutation carriers (per-allele OR?=?0.89, 95%CI 0.80-1.00, p?=?0.048), indicating a potential protective effect of this allele. These data suggest that that 6q22.33 confers a weak effect on breast cancer risk.     

Identification of Lactic Acid Bacteria from Chili Bo, a Malaysian Food Ingredient

Ninety-two strains of lactic acid bacteria (LAB) were isolated from a Malaysian food ingredient, chili bo, stored for up to 25 days at 28°C with no benzoic acid (product A) or with 7,000 mg of benzoic acid kg⁻¹ (product B). The strains were divided into eight groups by traditional phenotypic tests. A total of 43 strains were selected for comparison of their sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) whole-cell protein patterns with a SDS-PAGE database of LAB. Isolates from product A were identified as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus farciminis, Pediococcus acidilactici, Enterococcus faecalis, and Weissella confusa. Five strains belonging to clusters which could not be allocated to existing species by SDS-PAGE were further identified by 16S rRNA sequence comparison. One strain was distantly related to the Lactobacillus casei/Pediococcus group. Two strains were related to Weissella at the genus or species level. Two other strains did not belong to any previously described 16S rRNA group of LAB and occupied an intermediate position between the L. casei/Pediococcus group and the Weissella group and species of Carnobacterium. The latter two strains belong to the cluster of LAB that predominated in product B. The incidence of new species and subspecies of LAB in chili bo indicate the high probability of isolation of new LAB from certain Southeast Asian foods. None of the isolates exhibited bacteriocin activity against L. plantarum ATCC 14917 and LMG 17682.     

The role of demography,intra‐species variation,and species distribution models in species' projections under climate change

Organisms are projected to shift their distribution ranges under climate change. The typical way to assess range shifts is by species distribution models (SDMs), which predict species’ responses to climate based solely on projected climatic suitability. However, life history traits can impact species’ responses to shifting habitat suitability. Additionally, it remains unclear if differences in vital rates across populations within a species can offset or exacerbate the effects of predicted changes in climatic suitability on population viability. In order to obtain a fuller understanding of the response of one species to projected climatic changes, we coupled demographic processes with predicted changes in suitable habitat for the monocarpic thistle Carlina vulgaris across northern Europe. We first developed a life history model with species‐specific average fecundity and survival rates and linked it to a SDM that predicted changes in habitat suitability through time with changes in climatic variables. We then varied the demographic parameters based upon observed vital rates of local populations from a translocation experiment. Despite the fact that the SDM alone predicted C. vulgaris to be a climate ‘winner’ overall, coupling the model with changes in demography and small‐scale habitat suitability resulted in a matrix of stable, declining, and increasing patches. For populations predicted to experience declines or increases in abundance due to changes in habitat suitability, altered fecundity and survival rates can reverse projected population trends.     

Fast screening method for benzophenanthridine alkaloids in plant cell cultures of California poppy using a dansyl-glycine-β-cyclodextrin molecular recognition system

The association constant K for the host-guest complex comprised of sanguinarine/dansyl-glycine--cyclodextrin was found to be 5.75 × 105 M. The sensitivity factor(), which is defined as the ratio of the change of the fluorescence intensity after complex formation to the intensity of free, uncomplexed dansyl-glycine--cyclodextrin, showed an almost linear dependence on analyte concentration up to 50M sanguinarine. This molecular recognition system can be exploited to rapidly estimate the total concentration of benzophenanthridine alkaloids in suspension cultures of Eschscholtzia californica. © Rapid Science Ltd. 1998     

RNA helicase-mediated regulation of snoRNP dynamics on pre-ribosomes and rRNA 2′-O-methylation

RNA helicases play important roles in diverse aspects of RNA metabolism through their functions in remodelling ribonucleoprotein complexes (RNPs), such as pre-ribosomes. Here, we show that the DEAD box helicase Dbp3 is required for efficient processing of the U18 and U24 intron-encoded snoRNAs and 2′-O-methylation of various sites within the 25S ribosomal RNA (rRNA) sequence. Furthermore, numerous box C/D snoRNPs accumulate on pre-ribosomes in the absence of Dbp3. Many snoRNAs guiding Dbp3-dependent rRNA modifications have overlapping pre-rRNA basepairing sites and therefore form mutually exclusive interactions with pre-ribosomes. Analysis of the distribution of these snoRNAs between pre-ribosome-associated and ‘free’ pools demonstrated that many are almost exclusively associated with pre-ribosomal complexes. Our data suggest that retention of such snoRNPs on pre-ribosomes when Dbp3 is lacking may impede rRNA 2′-O-methylation by reducing the recycling efficiency of snoRNPs and by inhibiting snoRNP access to proximal target sites. The observation of substoichiometric rRNA modification at adjacent sites suggests that the snoRNPs guiding such modifications likely interact stochastically rather than hierarchically with their pre-rRNA target sites. Together, our data provide new insights into the dynamics of snoRNPs on pre-ribosomal complexes and the remodelling events occurring during the early stages of ribosome assembly.