Structures of common AHLs synthesized by <i>P aeruginosa.</i>

<p>Structures of the most common AHL quorum sensing compounds synthesized by <i>Pseudomonas aeruginosa</i> (C₄-HSL, and 3-oxo-C₁₂-HSL) and those used in this study (3-oxo-C₁₄-HSL, 3-oxo-C₁₂-HSL, COOH-3-oxo-C₁₂-HSL and 3-oxo-C₁₀-HSL).</p

A strong negative correlation is observed between peripheral mononuclear cell proliferation and AHL lipophilicity.

<p>The IC₅₀ for human peripheral mononuclear cell proliferation (PBMCs) isolated from three donors (* denotes n = 2) in the presence of AHLs of chain lengths between 8 and 14 carbons (values obtained from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013522#pone.0013522-Chhabra1" target="_blank">[18]</a>) exhibit a strong negative correlation (Pearsons r = −0.895, <i>p</i> = 0.016) against the predicted octanol/water partition coefficient (LogP) for each compound (calculated using ACD/i-Lab LogP algorithm 12.0). n = 3, ±SEM. Projected LogP are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013522#pone-0013522-t001" target="_blank">Table 1</a>.</p

Binding profiles of the interactions of AHLs with Lymphocyte membranes.

<p>Binding profiles of 3-oxo-C₁₄-HSL (Δ, hyperbolic), 3-oxo-C₁₂-HSL (•, sigmoidal) and 3-oxo-C₁₀-HSL (×, neither) on titration to di-8-ANEPPS labeled T-Lymphocytes (40,000 cells/ml) at 37°C normalised to DMSO controls. Profiles were fitted to simple hyperbolic and sigmoidal binding models (equations 1 and 2) and F-Tests were used to determine the best fitting model. In each experiment n = 3, ±SEM.</p

The interactions of AHLs with artificial membrane systems perturbs the membrane dipole potential.

<p>Fluorescence difference spectra obtained by subtracting di-8-ANEPPS excitation spectra (λem = 590 nm) of PC(100%) [A] or PC(70%)Cholesterol(30%) [B] membrane vesicles (400 µM) from those obtained after these membranes were exposed to the following QS molecules; 65 µM 3-oxo-C₁₄-HSL (thick dashed line), 200 µM 3-oxo-C₁₂-HSL (solid black line) and 200 µM 3-oxo-C₁₀-HSL (thin dashed and dotted line). Before subtraction, each spectrum was normalized to the integrated areas so that the difference spectra would reflect only the spectral shifts. Each difference spectrum was then normalised to a DMSO control (grey line) and a three point moving average applied to reduce noise. In all experiments the dye concentration was 10 µM and temperature was maintained at 37°C. [C] A dual wavelength ratiometric measurement of the dipole potential variation in di-8-ANEPPS. Additions of 22 µM 3-oxo-C_14-HSL or equivalent volumes of DMSO were made to 400 µM PC(100%). Samples were excited at 460 nm and 520 nm. Emission was read at 590 nm and the ratio <i>R</i>(460/520) was calculated (shown). All experiments n = 3.</p

Appendix A. Modeling the second definition of nonoverlapping chains.

Modeling the second definition of nonoverlapping chains

Complete database Bombus decline

The complete database, for GLOBAL DECLINE OF BUMBLEBEES IS PHYLOGENETICALLY STRUCTURED AND INVERSELY RELATED TO SPECIES RANGE SIZE AND PATHOGEN INCIDENCE, including potential variables associated with decline, described in the main text

Supplementary Material from Global decline of bumblebees is phylogenetically structured and inversely related to species range size and pathogen incidence

Conservation biology can profit greatly from incorporating a phylogenetic perspective into analyses of patterns and drivers of species extinction risk. We applied such approach to analyse patterns of bumblebee (<i>Bombus</i>) decline. We assembled a database representing approximately 43% of the approximately 260 globally known species, which included species extinction risk assessments following IUCN Red List categories and criteria, and information on species traits presumably associated with bumblebee decline. We quantified the strength of phylogenetic signal in decline, range size, tongue length and parasite presence. Overall, about one-third of the assessed bumblebees are declining and declining species are not randomly distributed across the <i>Bombus</i> phylogeny. Susceptible species were overrepresented in the subgenus <i>Thoracobombus</i> (approx. 64%) and underrepresented in the subgenus <i>Pyrobombus</i> (approx. 6%). Phylogenetic logistic regressions revealed that species with small geographical ranges and those in which none of three internal parasites were reported (i.e. <i>Crithidia bombi</i>, <i>Nosema</i> spp. or <i>Locustacarus buchneri</i>) were particularly vulnerable. Bumblebee evolutionary history will be deeply eroded if most species from threatened clades, particularly those stemming from basal nodes, become finally extinct. The habitat of species with restricted distribution should be protected and the importance of pathogen tolerance/resistance as mechanisms to deal with pathogens needs urgent research

Popat_et_al_data

Popat_et_al_dat

BiofilmCheatingData

BiofilmCheatingDat