end point was to detect new diagnoses of cardiovascular disease, we felt that these exclusion criteria were necessary to calculate the incidence of disease. Patient adherence cannot be assessed from pharmacy records. Other medications, family history, or lifestyle variables were not included in our analysis. All patients who were prescribed a tetracycline, regardless of duration, were included. It is not possible to know whether the prescription was truly given for acne/rosacea versus an infection. There are data that rosacea itself may increase the risk of cardiovascular disease (Duman et al., 2013), and it is possible that any indication for a tetracycline may perhaps positively affect the pro-inflammatory state of the rosacea patient. In private practice, the use of low-dose (20 or 40 mg daily) doxycycline is commonplace in order to take advantage of the sub-antimicrobial dose that also provides an anti-inflammatory effect in rosacea. This formulation is not available at our facility, and most patients were prescribed between 50-200 mg daily. Owing to sampling issues (three different medications in two different groups of patients, acne, and rosacea), we did not analyze the odds ratio for each of the tetracycline medications: tetracycline, doxycycline, and minocycline. Finally, as our veteran population was predominantly male, our results may not be generalizable to a more heterogeneous population. CONFLICT OF INTEREST JD has received fees from Galderma Laboratories for consulting that was performed after the design and completion of this study.

Calpastatin is an endogenous specific inhibitor of calpain, a calcium-dependent cysteine protease. Here we show that loss-of-function mutations in calpastatin (CAST) are the genetic causes of an autosomal-recessive condition characterized by generalized peeling skin, leukonychia, acral punctate keratoses, cheilitis, and knuckle pads, which we propose to be given the acronym PLACK syndrome. In affected individuals with PLACK syndrome from three families of different ethnicities, we identified homozygous mutations (c.607dup, c.424A>T, and c.1750delG) in CAST, all of which were predicted to encode truncated proteins (p.Ile203Asnfs∗8, p.Lys142∗, and p.Val584Trpfs∗37). Immunohistochemistry shows that staining of calpastatin is reduced in skin from affected individuals. Transmission electron microscopy revealed widening of intercellular spaces with chromatin condensation and margination in the upper stratum spinosum in lesional skin, suggesting impaired intercellular adhesion as well as keratinocyte apoptosis. A significant increase of apoptotic keratinocytes was also observed in TUNEL assays. In vitro studies utilizing siRNA-mediated CAST knockdown revealed a role for calpastatin in keratinocyte adhesion. In summary, we describe PLACK syndrome, as a clinical entity of defective epidermal adhesion, caused by loss-of-function mutations in CAST.

Our data suggest that the MTBDRplus line-probe assay is capable of detecting drug resistance for the CSF samples that have a PCR-positive result. We recommend PCR-based diagnosis and drug resistance test as routine assays for patients with suspected TBM.