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At day 18 in vitro (DIV18), we stimulated the neurons using either field stimulation (3-s trains consisting of 32 1-ms 50-mA pulses at 20 Hz for a total of 1 or 5 min), or via replacement of culture medium with medium of identical composition (this mechanically stimulates the cultures and also provides a fresh source of glutamate)

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Despite the widely promulgated risks associated with cigarette smoking in CD, more incisive data are required to further elucidate the actual relationship between smoking and disease pathways, while accounting for the several negative cofactors prevalent in smokers which cast uncertainty on the magnitude of the direct effect of smoking on disease pathophysiology and the efficacy of therapy