Taken together, our results indicate that although F-box gene gain and loss events do not occur as frequently in Euarchontoglires as they do in plants, the evolutionary pattern of 
the F-box gene family in these species is consistent with the birth-and-death evolution model. The protein structure may only be conserved in some parts of an active site responsible for catalysis, while the remaining peripheral regions may have changed considerably, causing change in substrate specificity. Indeed, the analysis of selective pressure at individual sites showed that the vast majority of functional domains are rich in residues with low v, whereas the residues in the core portion of the protein–protein interface underwent excess amino-acid fixation during the course of Euarchontoglires evolution. These results are consistent with previous findings in other organisms. In plants, while the F-box domain appears rather stable, some C-terminal protein-protein interaction domains such as Kelch and FBA show strong signatures of positive selection. Our results are also in agreement with previous findings in nematodes. Co-evolution of the substrate and Fbox protein interface may PR-957 explain the apparent fast evolution of the substrate-binding domain. Alternatively, mutation of residues that are solvent-exposed to a structural fold may be more tolerated than those located at the highly structured core. Therefore, further extensive research is required before the cause of this positive selection can be definitively determined. This study provides evidence that the cannabigerol quinone derivative, VCE-003, suppresses in vitro T cell responses, that it dampens the pro-inflammatory cytokine production by IL-17stimulated macrophages and attenuates the pathology development of EAE induced by MOG immunization. This improvement is evident through several measures of EAE pathology, including: a decreased neurological deficit score; reduced inflammatory cell infiltration, in particular that of CD4 cells; demyelination and axonal damage in the spinal cord. The mechanisms underlying the improvement in EAE induced by VCE-003 treatment are not clear, although the suppression of immune and inflammatory cell activity seems to be involved. In accordance with its BAY 43-9006 284461-73-0 pharmacological profile, the activation of both PPARc and CB2 receptors appear to be implicated in the amelioration of EAE by VCE-003. Microglia/macrophages play a dual role in the pathogenesis of MS as they contribute to lesion formation and axonal damage, but they also support repair mechanisms. The activation of microglia has been closely associated with the development of histopathological lesions, and the progression of MS and EAE. When microglia/ macrophages are activated they release some cytotoxic mediators that may provoke tissue injury, including pro-inflammatory cytokines like IL-1b, IFNc, IL-6, TNFa, nitric oxide and ROS. However, most importantly, several cytokines can reactivate microglia, which leads to feed-forward regulation of inflammation. The decrease in Iba-1 immunoreactivity following VCE-003 treatment reflected the reduced microglial activation in EAE mice. This observation suggests that VCE-003 might ameliorate EAE symptoms by dampening microglial activation in the spinal cord, thereby impairing important inflammatory steps that lead to the damage of myelin and axons.