No abstract

-cAMP is a highly regulated secondary messenger involved in many biological processes. Chronic activation of the cAMP pathway by catecholamines results in cardiac hypertrophy and fibrosis; however, the mechanism by which elevated cAMP leads to cardiomyopathy is not fully understood. To address this issue, we increased intracellular cAMP levels in HL-1 cardiomyocytes, a cell line derived from adult mouse atrium, using either the stable cAMP analog N 6 ,2=-O-dibutyryladenosine 3=,5=-cyclic monophosphate (DBcAMP) or phosphodiesterase (PDE) inhibitors caffeine and theophylline. Elevated cAMP levels increased cell size and altered expression levels of cardiac genes and micro-RNAs associated with hypertrophic cardiomyopathy (HCM), including Myh6, Myh7, Myh7b, Tnni3, Anp, Bnp, Gata4, Mef2c, Mef2d, Nfatc1, miR208a, and miR208b. In addition, DBcAMP altered the expression of DNA methyltransferases (Dnmts) and Tet methylcytosine dioxygenases (Tets), enzymes that regulate genomic DNA methylation levels. Changes in expression of DNA methylation genes induced by elevated cAMP led to increased global DNA methylation in HL-1 cells. In contrast, inhibition of DNMT activity with 5-azacytidine treatment decreased global DNA methylation levels and blocked the increased expression of several HCM genes (Myh7, Gata4, Mef2c, Nfatc1, Myh7b, Tnni3, and Bnp) observed with DBcAMP treatment. These results demonstrate that cAMP induces cardiomyocyte hypertrophy and altered HCM gene expression in vitro and that DNA methylation patterns mediate the upregulation of HCM genes induced by cAMP. These data identify a previously unknown mechanism by which elevated levels of cAMP lead to increased expression of genes associated with cardiomyocyte hypertrophy.

A determination of the dynamical evolution of the asteroid belt is difficult because the asteroid belt has evolved since the time of asteroid formation through mechanisms that include: (1) catastrophic collisions, (2) rotational disruption, (3) chaotic orbital evolution and (4) orbital evolution driven by Yarkovsky radiation forces. The timescales of these loss mechanisms are uncertain and there is a need for more observational constraints. In the inner main belt, the mean size of the non-family asteroids increases with increasing inclination. Here, we use that observation to show that all inner main belt asteroids originate from either the known families or from ghost families, that is, old families with dispersed orbital elements. We estimate that the average age of the asteroids in the ghost families is a factor of 1/3 less than the Yarkovsky orbital evolution timescale. However, this orbital evolution timescale is a long-term average that must allow for the collisional evolution of the asteroids and for stochastic changes in their spin directions. By applying these constraints on the orbital evolution timescales to the evolution of the size-frequency distribution of the Vesta asteroid family, we estimate that the age of this family is greater than 1.3 Gyr and could be comparable with the age of the Solar system. By estimating the number of ghost families, we calculate that the number of asteroids that are the root sources of the meteorites and the near-Earth asteroids that originate from the inner main belt is about 20.