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In all four models, protein citrullination was decreased, PAD activities were decreased, disease was attenuated and remyelination was observed.
White matter from PBS-treated mice had elevated PAD activities, which were normalized after 2CA (Fig. 8B).
The pre-policy data collection also assessed current levels of participation in Performance Appraisal and Development (PAD) activities.
Spinal cord PAD activities of PBS-treated mice reached threefold elevations, but reductions to normal levels were once again observed following treatment (Fig. 8C).
PAD activities indeed showed a sharp rise at disease onset, peaking at the height of the initial disease phase and declining to still above-background levels thereafter.
In order to place protein hypercitrullination and elevated PAD activities in the important role of the pathogenesis in MS, we sought to establish that increased PAD activity preceded disease, which we demonstrated in the ND4 model of MS (Moscarello et al., 2002).
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Alternatively, localization of PAD2 to the vicinity of intracellular calcium channels could promote the transient activation of PAD2 upon channel opening as the localized calcium concentrations are thought to reach the millimolar levels required to activate PAD activity.
Depending on the methylation signal, i.e., activating or inhibiting, PADs might act as a repressor if activating arginine methylation is inhibited or an activator if the balance of PAD activity is shifted toward the suppression of a repressive arginine methylation mark.
The cleaved form of AIF was indeed present in thapsigargin treated-cells, but was not detected when PAD activity was inhibited by Cl-amidine, indicating that PAD activation is required for the release of AIF from the mitochondria.
Although this compound shows limited utility as a cellular probe of PAD activity, its discovery does demonstrate that it is possible to identify potent non-active-site-targeted reversible PAD inhibitors.
Interestingly, we found that citrullination was limited to the arginine residues at 2, 8, and 17 on the histone H3 tail suggesting that these residues are the primary targets of PAD activity in mammary epithelial cells (Figure 4D).
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