Dose-dependent effects of peptide YY(3-36) on conditioned taste aversion in rats

We used a conditioned taste aversion test to assess whether PYY(3-36) reduces food intake by producing malaise. Two-hour IV infusion of PYY(3-36) (8, 15, and 30 pmol/kg/min) at dark onset in non-food-deprived rats produced a dose-dependent inhibition of feeding and a conditioned aversion to the flavored chow paired with PYY(3-36) infusion. In food-deprived rats, PYY(3-36) at 2 and 4 pmol/kg/min inhibited intake of a flavored saccharin solution without producing conditioned taste aversion, whereas higher doses (8 and 15 pmol/kg/min) inhibited saccharin intake and produced taste aversion. These results suggest that anorexic doses of PYY(3-36) may produce a dose-dependent malaise in rats, which is similar to that reported for PYY(3-36) infusion in humans. Previous studies have shown that PYY(3-36) potently inhibits gastric emptying, and that gut distention can produce a conditioned taste aversion. Thus, PYY(3-36) may produce conditioned taste aversion in part by slowing gastric emptying.     

CFTR regulates phagosome acidification in macrophages and alters bactericidal activity

Acidification of phagosomes has been proposed to have a key role in the microbicidal function of phagocytes. Here, we show that in alveolar macrophages the cystic fibrosis transmembrane conductance regulator Cl- channel (CFTR) participates in phagosomal pH control and has bacterial killing capacity. Alveolar macrophages from Cftr-/- mice retained the ability to phagocytose and generate an oxidative burst, but exhibited defective killing of internalized bacteria. Lysosomes from CFTR-null macrophages failed to acidify, although they retained normal fusogenic capacity with nascent phagosomes. We hypothesize that CFTR contributes to lysosomal acidification and that in its absence phagolysosomes acidify poorly, thus providing an environment conducive to bacterial replication.     

Vitamin D receptor signaling inhibits atherosclerosis in mice

Although vitamin D has been implicated in cardiovascular protection, few studies have addressed the role of vitamin D receptor (VDR) in atherosclerosis. Here we investigate the effect of inactivation of the VDR signaling on atherogenesis and the antiatherosclerotic mechanism of vitamin D. Low density lipoprotein receptor (LDLR)(-/-)/VDR(-/-) mice exhibited site-specific accelerated atherogenesis, accompanied by increases in adhesion molecules and proinflammatory cytokines in the aorta and cholesterol influx in macrophages. Macrophages showed marked renin up-regulation in the absence of VDR, and inhibition of renin by aliskiren reduced atherosclerosis in LDLR(-/-)/VDR(-/-) mice, suggesting that the renin-angiotensin system (RAS) promotes atherosclerosis in the absence of VDR. LDLR(-/-) mice receiving LDLR(-/-)/VDR(-/-) BMT developed larger lesions than LDLR(-/-) BMT controls. Moreover, LDLR(-/-) mice receiving Rag-1(-/-)/VDR(-/-) BMT, which were unable to generate functional T and B lymphocytes, still had more severe atherosclerosis than Rag-1(-/-) BMT controls, suggesting a critical role of macrophage VDR signaling in atherosclerotic suppression. Aliskiren treatment eliminated the difference in lesions between Rag-1(-/-)/VDR(-/-) BMT and Rag-1(-/-) BMT recipients, indicating that local RAS activation in macrophages contributes to the enhanced atherogenesis seen in Rag-1(-/-)/VDR(-/-) BMT mice. Taken together, these observations provide evidence that macrophage VDR signaling, in part by suppressing the local RAS, inhibits atherosclerosis in mice.     

Effects of hydrogen peroxide in a keratinocyte-fibroblast co-culture model of wound healing

Recently, there has been renewed interest in the role of reactive oxygen species (ROS), especially H(2)O(2), in wound healing. We previously showed that H(2)O(2) stimulates healing in a keratinocyte scratch wound model. In this paper, we used a more complex and physiologically relevant model that involves co-culturing primary keratinocytes and fibroblasts. We found that the two main cell types within the skin have different sensitivities to H(2)O(2) and to the widely used "antioxidant"N-acetyl-l-cysteine (NAC). Keratinocytes were very resistant to the toxicity of H(2)O(2) (250 and 500 μM) or NAC (5 mM). However, the viability of fibroblasts was decreased by both compounds. Using the co-culture model, we also found that H(2)O(2) increases re-epithelialization while NAC retards it. Our data further illustrate the possible role of ROS in wound healing and the co-culture model should be useful for screening agents that may influence the wound healing process.     

CFTR mediates bicarbonate-dependent activation of miR-125b in preimplantation embryo development

Although HCO₃⁻ is known to be required for early embryo development, its exact role remains elusive. Here we report that HCO₃⁻ acts as an environmental cue in regulating miR-125b expression through CFTR-mediated influx during preimplantation embryo development. The results show that the effect of HCO₃⁻ on preimplantation embryo development can be suppressed by interfering the function of a HCO₃⁻-conducting channel, CFTR, by a specific inhibitor or gene knockout. Removal of extracellular HCO₃⁻ or inhibition of CFTR reduces miR-125b expression in 2 cell-stage mouse embryos. Knockdown of miR-125b mimics the effect of HCO₃⁻ removal and CFTR inhibition, while injection of miR-125b precursor reverses it. Downregulation of miR-125b upregulates p53 cascade in both human and mouse embryos. The activation of miR-125b is shown to be mediated by sAC/PKA-dependent nuclear shuttling of NF-κB. These results have revealed a critical role of CFTR in signal transduction linking the environmental HCO₃⁻ to activation of miR-125b during preimplantation embryo development and indicated the importance of ion channels in regulation of miRNAs.     

Gametic models for multitrait selection schemes to study variance of response and drift under adverse genetic correlations

A major issue faced by breeders is how to effectively manage adverse correlations in breeding programs. We present results of a Monte Carlo allele-based simulation of the changes in response and variance of response under adverse genetic correlations by using the examples of two contrasting selection methods: the ‘Smith-Hazel’ selection index (SH) and independent culling (IC). We assumed several gene models, which included linkage and antagonistic pleiotropy as the primary drivers of adverse genetic correlations. The different behaviors of these selection methods allowed us to identify the mechanism behind the generation of uncertainty under antagonistic trait selection: IC had the properties of stabilizing selection, while SH behaved more similar to disruptive selection. Although SH outperformed IC in terms of genetic gain, this advantage happened at the cost of higher variance of response and loss of heterozygosity. Using an optimum selection algorithm (OS) to prevent the loss of heterozygosity through a constraint on inbreeding in SH/OS increased marginally the reliability, remaining still below that of IC under equal conditions. However, SH/OS had lower inbreeding (ΔF) than IC for equivalent levels of genetic gain, so a compromise between high selection reliability, low ΔF, and gain must be made by a breeder under antagonistic trait selection even with the use of optimization tools.     

Gain and diversity in advanced generation coastal Douglas-fir selections for seed production populations

Sublines are used in the third-generation breeding and testing of coastal Douglas-fir in British Columbia, with the original intent of selecting only one genotype per subline for production populations (e.g., seed orchards) to eliminate relatedness among parents (therein called “1/SL”). We evaluated three additional selection scenarios that did not consider the subline structure. One of the scenarios strictly selected on the basis of the highest breeding values of the trees (“TOP”); another scenario used the TOP selections, but assigned the number of ramets per selection proportionally to the selection breeding value (“LIND”); lastly, a simulated annealing technique was applied to maximize gain under explicit constraints on coancestry (“OPTS”). All three alternative selection scenarios resulted in some relatedness and coancestry among selections, but the last two provided increases in average breeding values compared to those obtained by the 1/SL scenario. Effective population sizes (and consequently inbreeding coefficients) varied among the three selection scenarios. Effects of the various selections on merchantable volume at rotation age were determined using a linear regression model based on an individual tree model (TASS), which was first run to determine the relationship between merchantable volume and inbreeding (f). LIND and TOP selections yielded the highest breeding values but, due to the increased coancestry among selections, paid a penalty in the merchantable volume determination. OPTS maximized merchantable volume at rotation age 60 after including more than 13 selections with an increase of around 3% over that obtained by the 1/SL selection scenario, with an associated increase in Ne of 50%. Other implications of the three alternative selection scenarios are discussed.