Termination and activation of store‐operated cyclic AMP production

Diverse pathophysiological processes (e.g. obesity, lifespan determination, addiction and male fertility) have been linked to the expression of specific isoforms of the adenylyl cyclases (AC1‐AC10), the enzymes that generate cyclic AMP (cAMP). Our laboratory recently discovered a new mode of cAMP production, prominent in certain cell types, that is stimulated by any manoeuvre causing reduction of free [Ca²⁺] within the lumen of the endoplasmic reticulum (ER) calcium store. Activation of this ‘store‐operated’ pathway requires the ER Ca²⁺ sensor, STIM1, but the identity of the enzymes responsible for cAMP production and how this process is regulated is unknown. Here, we used sensitive FRET‐based sensors for cAMP in single cells combined with silencing and overexpression approaches to show that store‐operated cAMP production occurred preferentially via the isoform AC3 in NCM460 colonic epithelial cells. Ca²⁺ entry via the plasma membrane Ca²⁺ channel, Orai1, suppressed cAMP production, independent of store refilling. These findings are an important first step towards defining the functional significance and to identify the protein composition of this novel Ca²⁺/cAMP crosstalk system.     

Nest‐site selection and productivity of Vesper Sparrows breeding in grazed habitats

ABSTRACT Livestock grazing in the shortgrass steppe of the Intermountain region of British Columbia may have a negative impact on ground‐nesting birds, but evidence of such an impact is lacking. We examined nest‐site selection and productivity of ground‐nesting Vesper Sparrows (Pooecetes gramineus) across sites with different grazing histories. From 2006 to 2008, we monitored Vesper Sparrow nests and measured vegetation characteristics known to be affected by grazing within nest patches. We used an information‐theoretic approach to test the relative importance of grazing‐affected vegetation variables as predictors of nest‐site selection, nest survival, and nestling condition. Vesper Sparrows selected nest sites with greater cover of late‐seral grass species that decrease in occurrence in response to grazing (i.e., “decreasers”) than was available in random patches in the same territories. Daily nest survival was also lower for nests surrounded by shorter vegetation (odds ratio = 1.12). However, “decreaser” cover was not associated with either of the two indices of productivity measured (daily nest survival probability and nestling condition). In addition, vegetation height, although an important driver of success, was not linked with nest‐site selection, and no vegetation‐cover variable was positively associated with productivity, despite nest concealment being central to our predictions. This suggests that predation risk for nests in areas with shorter vegetation was being elevated through some factor unrelated to concealment. Our results show that grazing reduced both the availability of suitable habitat for and nesting success of Vesper Sparrows, indicating that grazing could pose a threat to population persistence at a broader scale and could potentially contribute to observed declines. Additional research is needed to determine if grazing guidelines in the Intermountain region of British Columbia should be amended, better enforced, or both to prevent regional declines in populations of ground‐nesting grassland birds.     

A five-strain probiotic combination reduces pathogen shedding and alleviates disease signs in pigs challenged with Salmonella enterica Serovar Typhimurium

Salmonella spp. infection is a major cause of gastroenteritis, with many thousands of cases reported in the European Union every year. The use of probiotics offers the potential to improve this situation. Here, we investigate the effects of oral treatment of pigs with a defined lactic acid bacteria culture mixture on both clinical and microbiological signs of Salmonella enterica serovar Typhimurium infection. Fifteen weaned pigs blocked by sex and weight were administered control milk or a mixture of five probiotic strains as either a milk fermentate or milk suspension for a total of 30 days. The mixture consisted of two strains of Lactobacillus murinus and one strain each of Lactobacillus salivarius subsp. salivarius, Lactobacillus pentosus, and Pediococcus pentosaceous. Following probiotic administration for 6 days, animals were challenged orally with serovar Typhimurium; the health of the animals and the microbiological composition of their feces were monitored for 23 days postinfection. Animals treated with probiotic showed reduced incidence, severity, and duration of diarrhea. These animals also gained weight at a greater rate than control pigs administered skim milk. Mean fecal numbers of Salmonella were significantly reduced in probiotic-treated animals at 15 days postinfection (P = 0.01). The administered probiotic bacteria improved both the clinical and microbiological outcome of Salmonella infection. These strains offer significant benefit for use in the food industry and may have potential in human applications.     

Rapid Fluorescent-Antibody Stain Technique with Group A Streptococci

A rapid fluorescent-antibody stain technique used with young broth cultures of trypsinized group A streptococci is described. Results from 1,214 trypsinized cultures of group A streptococci employing the rapid stain method were equivalent, both in specificity and sensitivity, when compared to results obtained from identical non-trypsinized cultures stained with the longer standard technique of Cherry, Goldman, and Carski. Moreover, the rapid method requires less than 2 min to complete.     

Biosynthesis of the piperidine nucleus metabolism of dl-ϵ-N-METHYL-3H-lysine-2-14c by Sedum acre

dl-?-N-Methyl-³H-lysine-2-¹⁴C was prepared and administered to excised shoots of Sedum acre from which dl-sedamine was isolated. The ?-N-methyllysine was not incorporated without degradation and, therefore, it is probably not a direct precursor of the piperidine ring of sedamine.     

Identification and reciprocal introgression of a QTL affecting body mass in mice

The aim of this study was to examine the effects of a QTL in different genetic backgrounds. A QTL affecting body mass on chromosome 6 was identified in an F₂ cross between two lines of mice that have been divergently selected for this trait. The effect of the QTL on mass increased between 6 and 10 weeks of age and was not sex-specific. Body composition analysis showed effects on fat-free dry body mass and fat mass. To examine the effect of this QTL in different genetic backgrounds, the high body mass sixth chromosome was introgressed into the low body mass genetic background and vice versa by repeated marker-assisted backcrossing. After three generations of backcrossing, new F₂ populations were established within each of the introgression lines by crossing individuals that were heterozygous across the sixth chromosome. The estimated additive effect of the QTL on 10-week body mass was similar in both genetic backgrounds and in the original F₂ population (i.e., ~0.4 phenotypic standard deviations); no evidence of epistatic interaction with the genetic background was found. The 95% confidence interval for the location of the QTL was refined to a region of approximately 7 cM between D6Mit268 and D6Mit123.