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After centrifugation at 400 g for 6 min at 4 C, supernatants (cell-free BAL fluid) from the first two aliquots of 0.5 ml were stored at 80 C for cytokine analysis (ELISA), and cell pellets were used for flow-cytometry analysis

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Unfortunately, the psychological dependence often lasts longer than the physical one, and this could several months for the mental cravings to fade away