A molecular phylogeny of Glaphyridae (Coleoptera: Scarabaeoidea): evolution of pollination and association with ‘Poppy guild’ flowers

Among Scarabaeoidea, pollen feeding occurs in two major lineages, pleurostict Scarabaeidae and Glaphyridae. Here we infer for the first time the phylogeny of the scarabaeoid lineage Glaphyridae (Coleoptera) based on molecular data using partial gene sequences for 28S rRNA, cytochrome oxidase I (cox1) and 16S rRNA (rrnL) for 41 species. Based on the resulting tree topology, we inferred the timing of the origin of pollination and of their coevolution with different flower host taxa, with particular focus on the prominent red-coloured ‘poppy guild’ flowers. All genera of Glaphyridae that were sampled with multiple species were recovered as monophyletic. According to this analysis, the origin of Glaphyridae was around 140 Ma, while crown group divergence was dated to have occurred c. 112 Ma. Pollen feeding originated in Glaphyridae only once and much later than in other important pollinator groups, between 97 and 67 Ma. According to the reconstruction of ancestral feeding traits, Asteraceae (Cicharioidae) were the first hosts of Glaphyridae. Presumably, a further adaptive radiation was triggered by feeding on and pollination of red flowers (poppy guild) which arose at a later stage. It occurred for the first time between 30 and 40 Ma, whereby the clades that use red Ranunculaceae (Pygopleurus spp.) are older than clades using exclusively red Papaveraceae (Eulasia spp.) (25–30 Ma). The rather young age of red Ranunculaceae would imply that Pygopleurus species only subsequently used red Ranunculus species as flower hosts, and that a broad parallel host shift probably from red Papaver spp. to red Ranunculus asiaticus has occurred rather recently.     

Protein synthesis during the cell cycle of L5178Y cells

There are two peaks of ³H-leucine incorporation in the cell cycle of L5178Y cells. The first, during S stage, corresponds to a peak of ³H-leucine incorporation into the nuclear fraction. The second, during S or early G2, corresponds to a peak of ³H-leucine incorporation into the mitochondrial fraction. The rate of protein synthesis is unique for the proteins from each of the four fractions, nuclear, mitochondrial, microsomal, and soluble.The SDS polyacrylamide-gel electrophoretic patterns of ³H-leucine incorporation were different among three subcellular fractions: nuclear, mitochondrial, and microsomal + soluble. However, the incorporation pattern for each fraction remains qualitatively the same throughout the cell cycle.     

CD95 tyrosine phosphorylation is required for CD95 oligomerization

Proapoptotic stimuli, such as CD95 ligand and hydrophobic bile acids induce an epidermal growth factor receptor (EGFR)-catalyzed tyrosine phosphorylation of CD95-death receptor in hepatocytes, as a prerequisite for CD95-translocation to the plasma membrane, formation of the death-inducing signalling complex and execution of apoptotic cell death. However, the molecular role played by CD95 tyrosine phosphorylation remained unclear. The present study shows that CD95-tyrosine phosphorylation is required for CD95-oligomerization. Fluorescence resonance energy transfer (FRET)-analysis in Huh7 hepatoma cells, which were cotransfected with CD95-YFP/CD95-CFP revealed that stimulation of these cells with CD95 ligand, proapoptotic bile acids or hyperosmolarity resulted within 30 min in an intracellular FRET-signal, suggestive for CD95/CD95-oligomerization. After 120 min the FRET-signal was detected in the plasma membrane, indicating translocation of the CD95/CD95-oligomer to the plasma membrane. CD95/CD95-oligomerization was abolished in presence of AG1478 or a JNK-inhibitory peptide, i.e. maneuvers known to prevent EGFR-catalyzed CD95-tyrosine phosphorylation. Transfection studies with YFP/CFP-coupled CD95-mutants, which contain tyrosine/phenylalanine-exchanges in positions 232 and 291 (CD95^Y232,291F), revealed that at least one tyrosine (Y^232,291)-phosphorylated CD95 is required for CD95/CD95-oligomerization. FRET-studies in mouse embryonic fibroblasts, which in contrast to Huh7 express endogenous CD95, revealed that EGF, but not CD95L induced EGFR-homomerization, whereas CD95 ligand, but not EGF resulted in EGFR/CD95-heteromerization. These findings suggest that EGFR-catalyzed CD95-tyrosine phosphorylation is involved in the CD95/CD95-oligomerization process, which is induced by proapoptotic stimuli and is required for apoptosis induction.     

Smoking cessation and bronchial epithelial remodelling in COPD: a cross-sectional study

Background

Chronic Obstructive Pulmonary Disease (COPD) is associated with bronchial epithelial changes, including squamous cell metaplasia and goblet cell hyperplasia. These features are partially attributed to activation of the epidermal growth factor receptor (EGFR). Whereas smoking cessation reduces respiratory symptoms and lung function decline in COPD, inflammation persists. We determined epithelial proliferation and composition in bronchial biopsies from current and ex-smokers with COPD, and its relation to duration of smoking cessation.

Methods

114 COPD patients were studied cross-sectionally: 99 males/15 females, age 62 ± 8 years, median 42 pack-years, no corticosteroids, current (n = 72) or ex-smokers (n = 42, median cessation duration 3.5 years), postbronchodilator FEV₁ 63 ± 9% predicted. Squamous cell metaplasia (%), goblet cell (PAS/Alcian Blue⁺) area (%), proliferating (Ki-67⁺) cell numbers (/mm basement membrane), and EGFR expression (%) were measured in intact epithelium of bronchial biopsies.

Results

Ex-smokers with COPD had significantly less epithelial squamous cell metaplasia, proliferating cell numbers, and a trend towards reduced goblet cell area than current smokers with COPD (p = 0.025, p = 0.001, p = 0.081, respectively), but no significant difference in EGFR expression. Epithelial features were not different between short-term quitters (<3.5 years) and current smokers. Long-term quitters (≥3.5 years) had less goblet cell area than both current smokers and short-term quitters (medians: 7.9% vs. 14.4%, p = 0.005; 7.9% vs. 13.5%, p = 0.008; respectively), and less proliferating cell numbers than current smokers (2.8% vs. 18.6%, p < 0.001).

Conclusion

Ex-smokers with COPD had less bronchial epithelial remodelling than current smokers, which was only observed after long-term smoking cessation (>3.5 years).

Trial registration

NCT00158847     

Gross changes in reconstructions of historic land cover/use for Europe between 1900 and 2010

Historic land‐cover/use change is important for studies on climate change, soil carbon, and biodiversity assessments. Available reconstructions focus on the net area difference between two time steps (net changes) instead of accounting for all area gains and losses (gross changes). This leads to a serious underestimation of land‐cover/use dynamics with impacts on the biogeochemical and environmental assessments based on these reconstructions. In this study, we quantified to what extent land‐cover/use reconstructions underestimate land‐cover/use changes in Europe for the 1900–2010 period by accounting for net changes only. We empirically analyzed available historic land‐change data, quantified their uncertainty, corrected for spatial‐temporal effects and identified underlying processes causing differences between gross and net changes. Gross changes varied for different land classes (largest for forest and grassland) and led to two to four times the amount of net changes. We applied the empirical results of gross change quantities in a spatially explicit reconstruction of historic land change to reconstruct gross changes for the EU27 plus Switzerland at 1 km spatial resolution between 1950 and 2010. In addition, the reconstruction was extended back to 1900 to explore the effects of accounting for gross changes on longer time scales. We created a land‐change reconstruction that only accounted for net changes for comparison. Our two model outputs were compared with five commonly used global reconstructions for the same period and area. In our reconstruction, gross changes led in total to a 56% area change (ca. 0.5% yr^?1) between 1900 and 2010 and cover twice the area of net changes. All global reconstructions used for comparison estimated fewer changes than our gross change reconstruction. Main land‐change processes were cropland/grassland dynamics and afforestation, and also deforestation and urbanization.     

Dectin-1 stimulation induces suppressor of cytokine signaling 1, thereby modulating TLR signaling and T cell responses

Suppressor of cytokine signaling (SOCS) proteins serve as negative regulators of cytokine receptor signaling. However, SOCS proteins are not only induced via the JAK/STAT pathway, but are also transcribed on triggering of pattern recognition receptors such as TLRs. We now show that SOCS1 can also be induced by the non-TLR pattern recognition receptor Dectin-1 in murine bone marrow-derived dendritic cells and macrophages (BMMs). The C-type lectin Dectin-1 binds to yeasts and signals either in an autonomous manner or can be triggered in combination with TLRs. In our study, SOCS1 was expressed independently of any TLR engagement as a direct target gene of the Dectin-1 ligand Zymosan. Induction of SOCS1 was mediated by a novel pathway encompassing the tyrosine kinases Src and Syk that activated the downstream kinase proline-rich tyrosine kinase 2. Proline-rich tyrosine kinase 2, in turn, caused activation of the MAPK ERK, thereby triggering SOCS1 induction. SOCS1 did not modulate Dectin-1 signaling but affected TLR signaling, leading to decreased and abbreviated NF-κB activation in BMMs triggered by TLR9. Furthermore, IL-12 and IL-10 secretion were inhibited by SOCS1. We additionally observed that IL-17-producing Th cells were clearly increased by SOCS1 in BMMs. Our results show that SOCS1 is expressed via a new, NF-κB-independent pathway in Dectin-1-triggered murine BMMs and influences TLR cross talk and T cell priming.