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1 h prior to the assay time point, the medium was exchanged to the Seahorse XF Base Medium (Agilent Technologies) with 1 mM sodium pyruvate, 2 mM l-glutamine and 10 mM d-(+)-glucose (all Sigma-Aldrich), pH 7.4 and incubated at 37 C without CO 2

Participants self-reported their cigarette and e-cigarette dependence at one, three and six months using validated measures of dependence, including a Penn State-developed questionnaire that ranges from 0 (not at all dependent) to 20 (highly dependent)

This is why nicotine users often experience decay in unusual locations, like the roots of teeth or along the gumline, where saliva normally provides protection