Additional file 1 of Caffeine activates HOG-signalling and inhibits pseudohyphal growth in Saccharomyces cerevisiae

Additional file 1: Figure S1. Growth of yeast cells (strain BY4743) with (+ caf; caffeine concentration 10 mM) and without (-caf) caffeine. Growth was followed at 30 °C by microcultivation in 350 μl synthetic YNB medium in a Bioscreen C (automated shaking and optical density readings), with optical density (OD) measurements every 20 min. Inoculation to cultures from a small number of cells taken from an overnight colony on synthetic YNB agar plates (without caffeine). The growth analysis was performed in two independent replicates (n = 2) with in each case 5 technical replicates. Typical growth curves are displayed

Additional file 3 of Caffeine activates HOG-signalling and inhibits pseudohyphal growth in Saccharomyces cerevisiae

Additional file 3: Figure S3. Pseudohyphal growth in diploid cells was inhibited by caffeine. The diploid strain Σ1278 was cultured on solid SLAD media at 30 °C for 2 days. Upper panel control (water), lower panels: different concentration of caffeine (3, 5, 7, 10 mM). The edge of the colony (indicated by arrows) of the control shows filamentous extensions, while the edges of colonies grown in the presence of different concentrations of caffeine appeared smooth and even. A typical result is shown from two independent replicates, each containing two technical replicates for the control and 10 mM caffeine, while 3, 5 and 7 mM caffeine is a typical result from one run with two technical replicates. The original microscopic magnification for all images is 100×. However, in this supplementary figure selected portions of the images have been further enhanced (zoomed-in) to emphasis the colony edge-effects, resulting in another 10× magnification compared to the images presented in Figure 3 in the main text

Additional file 4 of Caffeine activates HOG-signalling and inhibits pseudohyphal growth in Saccharomyces cerevisiae

Additional file 4: Figure S4. Invasive growth under caffeine stress. Cells of the haploid Σ1278 strain were grown on YPD plates containing different concentrations of caffeine (3 and 10 mM). A typical result is shown from two independent replicates, each containing two technical replicates

Additional file 2 of Caffeine activates HOG-signalling and inhibits pseudohyphal growth in Saccharomyces cerevisiae

Additional file 2: Figure S2. Western blot analysis showing Hog1 dual-phosphorylation in response to caffeine. Exponentially growing wild type cells (BY4743) in synthetic medium without caffeine were treated with media with different concentrations of caffeine (10, 20, 30, 40 mM) for 5 min. Negative control was basal growth medium without any caffeine addition, and positive control is treatment with medium containing 500 mM NaCl for 5 min. A) Hog1-p detected with dualphosphorylated-specific antibody. B) Loading control of total Hog1 detected by Hog1 specific antibodies (non-phosphorylated form). Molecular weight standard to the left-most lane in both cases. A typical result is shown from two independent replicates

Adherence to a Mediterranean-like Diet as a Protective Factor Against COPD: A Nested Case-Control Study

A diet rich in nutrients has been suggested to have protective effects against the development of chronic obstructive pulmonary disease (COPD). Since the traditional Mediterranean diet is high in nutrients, including antioxidants, vitamins, and minerals, it is of interest to study as a protective factor against COPD. Our aim was therefore to study its associations with development of COPD using population-based prospective data from the Västerbotten Intervention Programme (VIP) cohort. Data on diet from 370 individuals, who later visited the Department of Medicine at the University Hospital, Umeå, Sweden, with a diagnosis of COPD, were compared to 1432 controls. Adherence to a Mediterranean diet was assessed by a modified version of the Mediterranean diet score (MDS). Cases were diagnosed with COPD 11.1 years (mean) (standard deviation [SD] 4.5 years) after first stating their dietary habits in the VIP at a mean age of 55.5 years (SD 6.6 years). Higher MDS was associated with a higher level of education and not living alone. After adjustment for co-habiting and education level, individuals with an intermediate MDS and those with the highest MDS had a lower odds of developing COPD (odds ratio [OR] 0.73, 95% confidence interval [CI] 0.56–0.95; OR 0.56, 95% CI 0.37–0.86, respectively). These results remained also after adjustment for smoking intensity, i.e., numbers of cigarettes smoked per day (OR 0.73, 95% CI 0.53–0.99; OR 0.59, 95% CI 0.35–0.97), respectively). To conclude, adherence to a Mediterranean-like diet seems to be inversely associated with the development of COPD.</p

Table_2_Osmoregulation in Barnacles: An Evolutionary Perspective of Potential Mechanisms and Future Research Directions.DOCX

Barnacles form a globally ubiquitous group of sessile crustaceans that are particularly common in the coastal intertidal. Several barnacle species are described as highly euryhaline and a few species even have the ability to colonize estuarine and brackish habitats below 5 PSU. However, the physiological and/or morphological adaptations that allow barnacles to live at low salinities are poorly understood and current knowledge is largely based on classical eco-physiological studies offering limited insight into the molecular mechanisms. This review provides an overview of available knowledge of salinity tolerance in barnacles and what is currently known about their osmoregulatory strategies. To stimulate future studies on barnacle euryhalinity, we briefly review and compare barnacles to other marine invertebrates with known mechanisms of osmoregulation with focus on crustaceans. Different mechanisms are described based on the current understanding of molecular biology and integrative physiology of osmoregulation. We focus on ion and water transport across epithelial cell layers, including transport mechanisms across cell membranes and paracellular transfer across tight junctions as well as on the use of intra- and extracellular osmolytes. Based on this current knowledge, we discuss the osmoregulatory mechanisms possibly present in barnacles. We further discuss evolutionary consequences of barnacle osmoregulation including invasion-success in new habitats and life-history evolution. Tolerance to low salinities may play a crucial role in determining future distributions of barnacles since forthcoming climate-change scenarios predict decreased salinity in shallow coastal areas. Finally, we outline future research directions to identify osmoregulatory tissues, characterize physiological and molecular mechanisms, and explore ecological and evolutionary implications of osmoregulation in barnacles.</p

The comparison of protein domain organisation of IR25a, IR8a and IR93a from <i>D</i>. <i>melanogaster</i>, <i>P</i>. <i>argus</i> and <i>B</i>. <i>improvisus</i>.

ATD amino-terminal domain (grey), Lig_chan Ligated ion channel L-glutamate- and glycine-binding site (PF10613) (violet) overlapping with PBPe (PF00060) domain (green). White boxes represent transmembrane domains predicted with TMHMM Server v. 2.0.</p

<i>De novo</i> antennular transcriptome assembly and annotation statistics, nt = nucleotides.

De novo antennular transcriptome assembly and annotation statistics, nt = nucleotides.</p

Alignment of PBPe domains from <i>D</i>. <i>melanogaster</i> IR25a, IR8a, IR93a and candidate <i>B</i>. <i>improvisus</i> IRs.

The three glutamate binding positions characterised from Benton et al. (2009) are marked with asterisk. S1 and S2 stand for ligand-binding lobes of the “Venus flytrap” of the iGluRs.</p

Table of TRP candidates identified from cyprid antennules transcriptome.

Table of TRP candidates identified from cyprid antennules transcriptome.</p