Osmotic Tolerance And Volume Regulation In In Vitro Cultures Of The Oyster Pathogen Perkinsus Marinus

Growth rate. cell size, osmotic tolerance, and volume regulation were examined in cells of Perkinsus marinus cultured in media of osmolalities ranging from 168 to 737 mOsm (6.5-27.0 ppt). Cells cultured at the low osmolalities of 168 and 256 mOsm (6.5 and 9.7 ppt) began log phase growth 4 days postsubculture, whereas cells cultured at the higher osmolalities 341, 433, and 737 mOsm (12.7. 16.0, and 27.0 ppt) began log phase growth 2 days postsubculture. During log phase growth, cells from the higher osmolalities 341, 433, and 737 mOsm had shorter doubling times than cells from the lower osmolalities 168 and 256 mOsm. During both log and stationary phase growth, the mean cell diameter of cells cultured at 168 mOsm was significantly greater than cells cultured at 341 and 737 mOsm; the mean diameters of cells cultured at 341 and 737 mOsm did not differ significantly from each other. P. marinus cells cultured in various osmolalities were exposed to artificial seawater treatments of 56-672 mOsm (2.5-24.7 ppt). After the hypoosmotic treatment of 56 mOsm, cells that had been cultured in medium of low osmolality, 168 mOsm, showed only 41% mortality whereas the cells from the 341-, 433-, and 737-mOsm culture groups experienced 100% mortality. During the hyperosmotic shock, all of the groups exhibited mortalities of less than 10%. In P. marinus cells cultured in medium of 737 mOsm and then placed in a 50% dilution, cell diameter increased 13% which was a volume increase of 44.5%, but cells returned to baseline size (size before osmotic shock) within 5 minutes. P. marinus cells cultured at low osmolalities can withstand both hypo- and hyperosmotic stress and use volume-regulatory mechanisms during hypoosmotic stress. Results suggest that transferring infected oysters to low salinity will result in strains of P. marinus acclimated to low salinity that will be able to withstand periodic events of extremely low salinity

Genome-wide meta-analysis identifies six novel loci associated with habitual coffee consumption.

Coffee, a major dietary source of caffeine, is among the most widely consumed beverages in the world and has received considerable attention regarding health risks and benefits. We conducted a genome-wide (GW) meta-analysis of predominately regular-type coffee consumption (cups per day) among up to 91 462 coffee consumers of European ancestry with top single-nucleotide polymorphisms (SNPs) followed-up in ~30 062 and 7964 coffee consumers of European and African-American ancestry, respectively. Studies from both stages were combined in a trans-ethnic meta-analysis. Confirmed loci were examined for putative functional and biological relevance. Eight loci, including six novel loci, met GW significance (log10Bayes factor (BF)>5.64) with per-allele effect sizes of 0.03-0.14 cups per day. Six are located in or near genes potentially involved in pharmacokinetics (ABCG2, AHR, POR and CYP1A2) and pharmacodynamics (BDNF and SLC6A4) of caffeine. Two map to GCKR and MLXIPL genes related to metabolic traits but lacking known roles in coffee consumption. Enhancer and promoter histone marks populate the regions of many confirmed loci and several potential regulatory SNPs are highly correlated with the lead SNP of each. SNP alleles near GCKR, MLXIPL, BDNF and CYP1A2 that were associated with higher coffee consumption have previously been associated with smoking initiation, higher adiposity and fasting insulin and glucose but lower blood pressure and favorable lipid, inflammatory and liver enzyme profiles (P<5 × 10-8).Our genetic findings among European and African-American adults reinforce the role of caffeine in mediating habitual coffee consumption and may point to molecular mechanisms underlying inter-individual variability in pharmacological and health effects of coffee