We investigated the efficacy of standard (8-week nicotine, 16-week placebo) vs extended (24-week nicotine) patch therapy for smoking cessation, and the effect of slow nicotine metabolism, indicated by CYP2A6 reduced metabolizer (RM) genotype or low 3-hydroxycotinine/cotinine ratio (3HC/COT), on abstinence. RM versus normal genotype predicted lower 3HC/COT. Extended vs standard treatment produced higher abstinence at 24 weeks (32% vs 20%), but not at 52 weeks (both 14%). Low 3HC/COT and RM genotype predicted higher abstinence on extended versus standard treatment at 24 (47% vs 25%, 38% vs 17%) and 28 weeks (34% vs 19%, 23% vs 11%), while high 3HC/COT or normal genotype did not. Within extended treatment, low versus high 3HC/COT predicted higher abstinence at 8 (48% vs 29%), 24 (47% vs 25%), and 28 weeks (34% vs 16%), with similar trends for the genotype effect. Overall, extending nicotine treatment increased abstinence during therapy, particularly for slow metabolizers.

We investigated the efficacy of standard (8-week nicotine, 16-week placebo) vs extended (24-week nicotine) patch therapy for smoking cessation, and the effect of slow nicotine metabolism, indicated by CYP2A6 reduced metabolizer (RM) genotype or low 3-hydroxycotinine/cotinine ratio (3HC/COT), on abstinence. RM versus normal genotype predicted lower 3HC/COT. Extended vs standard treatment produced higher abstinence at 24 weeks (32% vs 20%), but not at 52 weeks (both 14%). Low 3HC/COT and RM genotype predicted higher abstinence on extended versus standard treatment at 24 (47% vs 25%, 38% vs 17%) and 28 weeks (34% vs 19%, 23% vs 11%), while high 3HC/COT or normal genotype did not. Within extended treatment, low versus high 3HC/COT predicted higher abstinence at 8 (48% vs 29%), 24 (47% vs 25%), and 28 weeks (34% vs 16%), with similar trends for the genotype effect. Overall, extending nicotine treatment increased abstinence during therapy, particularly for slow metabolizers.

ATP-sensitive K+ channels (KATP) channels were first described in the cardiac muscles. KATP channels are a complex of regulatory sulphonylurea receptor subunits and pore-forming inward rectifier subunits such as Kir6.1. Nicotine, an exogenous substance, adversely affects cardiovascular function in humans. Acetylcholine (ACh) is well known as a key neurotransmitter of the parasympathetic nervous system. ACh effects are usually related to binding to muscarinic receptors and stimulating second messengers that relay and direct the extracellular signals to different intracellular destinations, resulting in modulated cellular activity. We hypothesize that nicotine and ACh may modulate Kir6.1 channels via different mechanisms. Using the whole cell patch-clamp technique, the interactions of nicotine and ACh with Kir6.1 subunit permanently expressed in Human Embryonic Kidney (HEK-293) cells as well as the underlying mechanisms were studied.<p> Non-transfected HEK-293 cells possess an endogenous K+ current with current density of 3.2 ± 1.4 pA/pF at 150 mV (n = 9). Stable expression of Kir6.1 subunits cloned from rat mesenteric artery in HEK-293 cells yielded a detectable inward rectifier KATP current (-23.9 ± 1.6 pA/pF at 150 mV, n = 6). In the presence of 0.3 mM ATP in the pipette solution, nicotine at 30 and 100 µM increased the expressed Kir6.1 currents by 42 ± 11.8 and 26.2 ± 14.6%, respectively (n = 4-6, p<0.05). In contrast, nicotine at 1-3 mM inhibited Kir6.1 currents (p<0.05). Nicotine at 100 µM increased the production of superoxide anion (O2.-) by 20.3 ± 5.7% whereas at 1 mM it significantly decreased the production of O2.- by 37.7 ± 4.3%. The hypoxanthine/xanthine oxidase (HX/XO) reaction was used as a source of O2.-. Co-application of HX and XO to the transfected HEK-293 cells resulted in a significant and reproducible increase in Kir6.1 currents. Tempol, a scavenger of O2.-, abolished the stimulatory effect of HX/XO on Kir6.1 currents. Tempol also abolished the stimulatory effect of 30 mM nicotine on Kir6.1 currents (-28.3 ± 6.1 pA/pF vs. -31.2 ± 7.3 pA/pF at -150 mV, n = 6-9 for each group, p>0.05). <p> In the presence of 0.3 mM ATP in the pipette solution, ACh concentration-dependently increased the expressed Kir6.1 currents. At 1 µM, ACh increased Kir6.1 currents from -19 ± 2.5 to 31.7 ± 2.1 pA/pF (n = 8, p < 0.05). Pretreatment of the transfected HEK-293 cells with either 2 or 20 µM atropine, 100 nM a-bungarotoxin, 100 µM mecamylamine, 2 µM prazosin, 1 µM propranolol, or 10 µM dihydro-b-erythroidine hydrobromide did not alter the stimulatory effect of ACh on Kir6.1 currents (n = 4 - 5 for each group, p<0.05). When intracellular ATP was increased to 5 mM, ACh at 10 µM still exhibited its stimulatory effect (-16.4 ± 2.3 to 25.5 ± 3.8 pA/pF, n = 8, p<0.05). For the first time, the present study provides an insight for the interactions of nicotine and ACh with Kir6.1 subunits. Our data demonstrate that micromolar concentration of nicotine and ACh stimulated Kir6.1 channels. Nicotine at millimolar concentrations inhibited Kir6.1 channels. The dual effect of nicotine, not mediated by nAChR, are mediated partially by O2.- levels in the cells. The ACh excitatory effect is mediated neither by an AChR-dependent mechanism, nor by alteration in ATP metabolism. This study challenges the traditional explanations for the receptor-mediated effects of nicotine and ACh on ion channels and opens a new door to understand the effects of nicotine and ACh on KATP channels in many cellular systems.

The methadone maintenance treatment population suffers from high rates of comorbid psychiatric and substance use disorders. Despite a more than 40-year treatment history, not all patients are satisfied with methadone treatment and more than half of the patients complain of significant inter-dose withdrawal at least some of the time. The objectives of this research were to investigate the pharmacological response to methadone under the influence of comorbid major depressive disorder and smoking; and to identify factors other than physical withdrawal symptoms that can differentiate patients based on their complaints of dissatisfaction with treatment. In Study 1, seven depressed methadone maintenance patients experienced more opioid withdrawal symptomatology over a 24-hour methadone-dosing interval than 10 nondepressed methadone patients. Depression severity was significantly correlated with trough opioid withdrawal severity. This suggests that depression or depressive symptoms are related to reported opioid withdrawal. In Study 2, many factors other than physical opioid withdrawal symptoms were able to differentiate patients who were satisfied with treatment (holders, n=25), partially satisfied with treatment (partial holders, n=35), and not satisfied with treatment(nonholders, n=30). Results suggested that these patient satisfaction groups cluster differently depending on physical opioid withdrawal, mood, psychological distress, and personality. Nonholders experienced more physical withdrawal symptoms, craving for opioids, and negative drug effects. Holders had less psychological distress and experienced less negative mood states than the other groups. Partial holders had less agreeable personalities compared to patients in the other groups. In Study 3, opioid and nicotine withdrawal symptoms and effects were measured in 40 methadone-maintained patients who were current smokers during trough and peak methadone effects, both pre and post-nicotine administration. Cigarette smoking enhanced opioid withdrawal suppression during the peak methadone condition, methadone attenuated nicotine withdrawal, and methadone and nicotine shared many of the same main effects, suggesting that smoking and methadone effects may be inseparable dimensions. In summary, the results of these studies suggest that in addition to physical symptoms, mood related factors are important to opioid withdrawal perception and that the mood factors and drug interactions can impact on a patient’s perception of satisfaction with methadone treatment.

In Canada, nicotine replacement therapy (NRT) is a best practice for adult smoking cessation, but it is not recommended for use by youth smokers. Previous research has indicated that more than 20 percent of high school-aged smokers in Canada had used NRT, despite the cross-Canada requirement that youths under the age of 18 have a physician’s prescription to purchase NRT. The goal of this study was to examine both student and school-level characteristics associated with use of NRT by youths. Data from 29,296 grade 9 to 12 students who participated in the 2008-2009 National Youth Smoking Survey (YSS) were combined with Canadian census and built environment data in multilevel logistic regression models. The associations between lifetime and current NRT use with student characteristics (i.e., smoking status, social smoking connections) were examined alongside school environment factors such as urban/rural location and pharmacy density within a one kilometre radius of schools. In 2008-2009, 21.1% of youth smokers in Canada had ever used NRT and 5.1% were currently using NRT. Odds of NRT use were highest among daily smokers, boys, youths who had made multiple quit attempts, and youths who self-identified as smokers. Attending a school located within an urban area increased youths’ odds of NRT use, whereas higher density of pharmacies surrounding a school was inversely associated with NRT use. This study is the first to identify significant between school differences in NRT use. It also reveals that many youths are using NRT in the absence of a quit attempt. Further research is needed to identify school characteristics that impact NRT use, and understand how youths are accessing NRT.

ATP-sensitive K+ channels (KATP) channels were first described in the cardiac muscles. KATP channels are a complex of regulatory sulphonylurea receptor subunits and pore-forming inward rectifier subunits such as Kir6.1. Nicotine, an exogenous substance, adversely affects cardiovascular function in humans. Acetylcholine (ACh) is well known as a key neurotransmitter of the parasympathetic nervous system. ACh effects are usually related to binding to muscarinic receptors and stimulating second messengers that relay and direct the extracellular signals to different intracellular destinations, resulting in modulated cellular activity. We hypothesize that nicotine and ACh may modulate Kir6.1 channels via different mechanisms. Using the whole cell patch-clamp technique, the interactions of nicotine and ACh with Kir6.1 subunit permanently expressed in Human Embryonic Kidney (HEK-293) cells as well as the underlying mechanisms were studied.<p> Non-transfected HEK-293 cells possess an endogenous K+ current with current density of 3.2 ± 1.4 pA/pF at 150 mV (n = 9). Stable expression of Kir6.1 subunits cloned from rat mesenteric artery in HEK-293 cells yielded a detectable inward rectifier KATP current (-23.9 ± 1.6 pA/pF at 150 mV, n = 6). In the presence of 0.3 mM ATP in the pipette solution, nicotine at 30 and 100 µM increased the expressed Kir6.1 currents by 42 ± 11.8 and 26.2 ± 14.6%, respectively (n = 4-6, p<0.05). In contrast, nicotine at 1-3 mM inhibited Kir6.1 currents (p<0.05). Nicotine at 100 µM increased the production of superoxide anion (O2.-) by 20.3 ± 5.7% whereas at 1 mM it significantly decreased the production of O2.- by 37.7 ± 4.3%. The hypoxanthine/xanthine oxidase (HX/XO) reaction was used as a source of O2.-. Co-application of HX and XO to the transfected HEK-293 cells resulted in a significant and reproducible increase in Kir6.1 currents. Tempol, a scavenger of O2.-, abolished the stimulatory effect of HX/XO on Kir6.1 currents. Tempol also abolished the stimulatory effect of 30 mM nicotine on Kir6.1 currents (-28.3 ± 6.1 pA/pF vs. -31.2 ± 7.3 pA/pF at -150 mV, n = 6-9 for each group, p>0.05). <p> In the presence of 0.3 mM ATP in the pipette solution, ACh concentration-dependently increased the expressed Kir6.1 currents. At 1 µM, ACh increased Kir6.1 currents from -19 ± 2.5 to 31.7 ± 2.1 pA/pF (n = 8, p < 0.05). Pretreatment of the transfected HEK-293 cells with either 2 or 20 µM atropine, 100 nM a-bungarotoxin, 100 µM mecamylamine, 2 µM prazosin, 1 µM propranolol, or 10 µM dihydro-b-erythroidine hydrobromide did not alter the stimulatory effect of ACh on Kir6.1 currents (n = 4 - 5 for each group, p<0.05). When intracellular ATP was increased to 5 mM, ACh at 10 µM still exhibited its stimulatory effect (-16.4 ± 2.3 to 25.5 ± 3.8 pA/pF, n = 8, p<0.05). For the first time, the present study provides an insight for the interactions of nicotine and ACh with Kir6.1 subunits. Our data demonstrate that micromolar concentration of nicotine and ACh stimulated Kir6.1 channels. Nicotine at millimolar concentrations inhibited Kir6.1 channels. The dual effect of nicotine, not mediated by nAChR, are mediated partially by O2.- levels in the cells. The ACh excitatory effect is mediated neither by an AChR-dependent mechanism, nor by alteration in ATP metabolism. This study challenges the traditional explanations for the receptor-mediated effects of nicotine and ACh on ion channels and opens a new door to understand the effects of nicotine and ACh on KATP channels in many cellular systems.

Our study investigated the potential for a combination of diabetes and nicotine treatment to affect blood pressure in the rat. We used streptozotocin injection and oral nicotine feeding as models of type-1 diabetes and smoking respectively. Blood pressure was assessed using the indirect tail-cuff technique. In an attempt to further characterize our experimental model, we also observed body weight, plasma glucose and the contractility of aortic segments in various treatment groups. Our data was expressed as mean ± SEM, and significance was regarded as P < 0.05. We found that a combination of streptozotocin and nicotine treatment resulted in a significant elevation of systolic blood pressure compared with either treatment alone, or control. Furthermore, assessment of aortic contractility showed alteration of reactivity to both phenylephrine and sodium nitroprusside as a result of the combination treatment. We also observed a trend for our combination treatment to exacerbate the elevation of plasma glucose level seen in streptozotocin induced diabetic rat models. This study serves as an experimental basis to underline the importance of cessation of tobacco use for individuals with diabetes mellitus.

Thesis (M. Phil.)--University of Hong Kong, 2001. / Includes bibliographical references (leaves 112-126).

Thesis (Ph. D.)--Ohio State University, 2009. / Title from first page of PDF file. Includes bibliographical references (p. 153-165).

Little is currently known about the ability of electronic cigarettes to manage tobacco withdrawal symptoms and their abuse liability. In the current study eight conventional cigarette smokers completed nine within-subject study sessions. In the first session participants practiced using an electronic cigarette containing 16 mg/ml of nicotine over six 10-puff bouts. Remaining study sessions were comprised of four two-day blocks (one for each condition), which assessed measures of tobacco withdrawal symptoms and abuse liability following unrestricted cigarette smoking and 24-hour tobacco deprivation. Study conditions included an electronic cigarette with 0, 8, or 16 mg/ml nicotine concentrations, or preferred brand of conventional cigarette. Following 24-hours of tobacco deprivation, smoking conventional cigarettes ameliorated many of the self-report and physiological symptoms (decreased heart rate) associated with tobacco deprivation, while no attenuation of withdrawal symptoms was indicated following using electronic cigarettes, independent of nicotine dose. On abuse liability measures there were no significant changes following using an electronic cigarette (regardless of nicotine concentration), while conventional cigarettes engendered significant changes on abuse liability measures. Within the conditions of this study, first-generation electronic cigarettes had no measurable efficacy in ameliorating tobacco withdrawal symptoms and a reduced abuse liability compared to conventional tobacco cigarettes.