The MAP kinase pathways are involved in the sensitization of <i>S. cerevisiae</i> to C2-phytoceramide.

<p>Survival of wild type W303-1A cells and of mutant strains after exposure to 30 µM C2-phytoceramide for 120 min. (<i>ssk1</i>Δ, P<0.0001, <i>sln1</i>Δ, P<0.0001 <i>hog1</i>, P<0.01 and <i>ste20</i>Δ, P<0.05), (<i>ste3</i>Δ, P<0.05), One-Way ANOVA. Data are given as mean ± SE of at least 3 independent experiments, with 5 replicas in each experiment.</p

Loss of cell viability is rescued by ergosterol biosynthesis inhibitors, amphotericin B and methyl-β-cyclodextrin.

<p>(<b>A</b>) Survival of W303-1A cells exposed to amphotericin B 1 µg/ml, ketoconazole 300 µM, clotrimazole 300 µM or methyl-β-cyclodextrin 5 mg/ml for 30 min followed by an incubation with C2-phytoceramide for 120 min. Values represent mean ± SE of at least 3 independent experiments, with 5 replicas in each experiment. (<b>B</b>) Survival of wild type W303-1A and <i>rvs161</i>Δ cells exposed to 30 µM C2-phytoceramide or 30 µM C2-ceramide. CFU values of the C2-phytoceramide-treated mutant cells are significantly different from wild type-treated cells P<0.01, One-Way ANOVA.</p

<i>S. cerevisiae</i> cells in G2/M are more sensitive to C2-phytoceramide.

<p>(<b>A</b>) Survival of W303-1A cells synchronized in nitrogen starvation medium. Cells were transferred to SC gal medium and then exposed to 30 µM C2-phytoceramide (▼-dashed lines) or equivalent volume of solvent (■), or maintained in nitrogen starvation medium and exposed to 30 µM C2-phytoceramide (∇-dashed lines) or equivalent volume of solvent (□). CFU values of cells transferred to SC gal and treated with 30 µM C2-phytoceramide are significantly different from C2-phytoceramide treated cells kept in nitrogen starvation medium and from DMSO-treated cells transferred or not to SC Gal, for all time points, P<0.001, Two-Way ANOVA. All CFU values represent mean ± SE of at least 3 independent experiments, with 5 replicas in each experiment. (<b>B</b>) Cell cycle progression of the cells described in (A). Data represent mean ± SE of at least 3 independent experiments.</p

<i>S. cerevisiae</i> cells are sensitive to ceramides.

<p>(<b>A</b>) Survival of W303-1A cells exposed to 30 µM C2-phytoceramide (∇), 30 µM C6-ceramide (*), 30 µM C2-ceramide (▼), or equivalent volume of solvent (■). CFU values of C2-treated cells significantly different from DMSO-treated cells, P<0.0001, Two-Way ANOVA. (<b>B</b>) Survival of W303-1A cells exposed to 10 µM (●), 20 µM (○), 30 µM (∇-dashed lines) and 40 µM (♦-dashed lines) C2-phytoceramide, or equivalent volume of solvent (■). CFU values of C2-phytoceramide-treated cells (30 µM or 40 µM) are significantly different from DMSO-treated cells for all time points, P<0.001, Two-Way ANOVA. All CFU values (A and B) represent mean ± SE of at least 3 independent experiments, with 5 replicas in each experiment. (<b>C</b>) Cell cycle progression of cells exposed to 30 µM C2-phytoceramide or equivalent volume of solvent. Data from a representative experiment (of 3 independent experiments) is shown.</p

Distribution of sterol-rich domains in <i>S. cerevisiae</i> cells exposed to C2-phytoceramide.

<p>(<b>A</b>) Fluorescence microscopy pictures of W303-1A cells exposed to 30 µM C2-phytoceramide, 40 µM C2-ceramide, 5 mg/ml methyl-®-cyclodextrin or 0.1% DMSO for 120 min and stained with filipin (5 mg/ml). (<b>B</b>) Percentage of yeast cells with ergosterol displacement. Cells were treated as described in (<b>A</b>) and the number of cells with ergosterol displacement was determined by counting at least 120 cells per sample, in three independent experiments. P<0.01 respectively, One-Way ANOVA. (<b>C</b>) Percentage of yeast cells with perturbed Pma1p-GFP distribution. W303-1A cells were transformed with a single copy vector derived from pRS316 expressing Pma1p-GFP (3). At least 300 cells per sample were counted. P<0.01.</p

Cell death markers observed after exposure of <i>S. cerevisiae</i> to C2-phytoceramide.

<p>(<b>A</b>) Percentage of PI positive cells in W303-1A cultures exposed to C2-phytoceramide or to equivalent volume of solvent. Values represent mean ± SE of at least 3 independent experiments. Percentages of PI-stained cells of C2-phytoceramide-treated cells (30 µM and 40 µM) significantly different from DMSO-treated cells, P<0.001 for time 60 and 120 min and P<0.01 for time 180 min, Two-Way ANOVA. (<b>B</b>) Percentage of yeast cells with chromatin condensation, determined by counting at least 300 cells per sample after a 120 min treatment with 30 µM C2-phytoceramide or equivalent volume of solvent, in three independent experiments. <i>P</i><0.05 One-Way ANOVA.</p

Teaching open and reproducible scholarship: a critical review of the evidence base for current pedagogical methods and their outcomes

In recent years, the scientific community has called for improvements in the credibility, robustness and reproducibility of research, characterized by increased interest and promotion of open and transparent research practices. While progress has been positive, there is a lack of consideration about how this approach can be embedded into undergraduate and postgraduate research training. Specifically, a critical overview of the literature which investigates how integrating open and reproducible science may influence student outcomes is needed. In this paper, we provide the first critical review of literature surrounding the integration of open and reproducible scholarship into teaching and learning and its associated outcomes in students. Our review highlighted how embedding open and reproducible scholarship appears to be associated with (i) students' scientific literacies (i.e. students’ understanding of open research, consumption of science and the development of transferable skills); (ii) student engagement (i.e. motivation and engagement with learning, collaboration and engagement in open research) and (iii) students' attitudes towards science (i.e. trust in science and confidence in research findings). However, our review also identified a need for more robust and rigorous methods within pedagogical research, including more interventional and experimental evaluations of teaching practice. We discuss implications for teaching and learning scholarship

Teaching open and reproducible scholarship: a critical review of the evidence base for current pedagogical methods and their outcomes

In recent years, the scientific community has called for improvements in the credibility, robustness and reproducibility of research, characterized by increased interest and promotion of open and transparent research practices. While progress has been positive, there is a lack of consideration about how this approach can be embedded into undergraduate and postgraduate research training. Specifically, a critical overview of the literature which investigates how integrating open and reproducible science may influence student outcomes is needed. In this paper, we provide the first critical review of literature surrounding the integration of open and reproducible scholarship into teaching and learning and its associated outcomes in students. Our review highlighted how embedding open and reproducible scholarship appears to be associated with (i) students' scientific literacies (i.e. students’ understanding of open research, consumption of science and the development of transferable skills); (ii) student engagement (i.e. motivation and engagement with learning, collaboration and engagement in open research) and (iii) students' attitudes towards science (i.e. trust in science and confidence in research findings). However, our review also identified a need for more robust and rigorous methods within pedagogical research, including more interventional and experimental evaluations of teaching practice. We discuss implications for teaching and learning scholarship.</p