On testing Day 6, participants attended a 4-hr nicotine nasal spray laboratory session. During this session, participants used the nicotine nasal spray (2-mg dose at Time 0), and a series of eight-timed sellekchem blood samples were collected for serum nicotine analysis (Time: ?1, 5, 10, 20, 30, 45, 60, and 90min). Nicotine serum samples were later analyzed in batches at Hennepin County Medical Center Laboratories using gas chromatography with nitrogen phosphorus detection (Jacob, Wilson, & Benowitz, 1981). Intraassay and interassay coefficient of variations ranged between 2.7% and 5.6%, respectively. At each timepoint, participants completed the Subjective State Scale (al��Absi, Lovallo, McKey, & Pincomb, 1994; al��Absi et al., 1998) for measurement of positive (i.e., cheerful) and negative affect (i.

e., angry) using an 8-point scale. Blood pressure and heart rate were also measured via an automated blood pressure machine at each timepoint. Upon completion of the testing week, participants resumed ad libitum smoking until they arrived at the alternate menstrual phase (approximately 6 weeks later depending on cycle length). Identical data collection procedures were then completed. Participants were compensated for their time. All procedures were approved by the human subjects committee at the University of Minnesota. Analysis Maximal nicotine concentration (Cmax) was the highest observed nicotine concentration, and time to maximal nicotine concentration (Tmax) was the first timepoint at which this concentration occurred.

In instances when all of the concentrations for a given laboratory period were below the lower limit of quantification (which was 2ng/ml), a value of 1ng/ml was assigned as the Cmax and the 5-min timepoint was assigned as the Tmax. Descriptive statistics were calculated for demographics and baseline characteristics (mean and standard deviation for continuous variables, and count and percent for categorical variables). Change scores for heart rate, systolic blood pressure, and diastolic blood pressure were calculated by subtracting baseline values (Time ?1min) from the testing values (Time 5, 10, 20, 30, 45, 60, and 90min). Random-intercept models with variables for sequence (carry-over) and time effects were used to investigate the association of menstrual phase and depressive symptoms group with serum nicotine, heart rate, systolic blood pressure, and diastolic blood pressure.

In subsequent analyses, the depressive symptoms group-menstrual phase interactions were included in the models. To examine the effect of sex hormone values (estradiol, Dacomitinib progesterone, and progesterone/estradiol ratio) on serum nicotine, heart rate, systolic blood pressure, and diastolic blood pressure, the models were rerun replacing menstrual phase with the sex hormone value. The same analysis was done to assess the positive and negative affect outcomes.