Nancy J. Haley

Nicotine administration to rats: methodological considerations.

The effects of nicotine on normal physiological function are of increasing concern. Preliminary to studies on the effects of prenatal exposure to nicotine, we examined methods of administering nicotine to rats. Drinking water containing nicotine was not palatable to rats and was an unsatisfactory method in our hands, producing weight loss and large decreases in fluid intake. Administration of nicotine in a complete liquid diet produced better results but the data suggest that oral administration of nicotine may interfere with absorption of some nutrients. Osmotic mini-pumps were found to be the best mechanism of nicotine delivery of those tried. There were no significant effects on food or water intake nor on weight gain, particularly when using a short term anesthetic for pump implantation. Plasma nicotine and cotinine levels were directly correlated to dose of nicotine delivered. Plasma nicotine levels similar to levels reported in humans were obtained.

Smoking and cervical intraepithelial neoplasia: Nicotine and cotinine in serum and cervical mucus in smokers and nonsmokers

A blood sample before treatment was taken from 35 women with cervical intraepithelial neoplasia. Levels of nicotine and cotinine were analyzed by radioimmunoassay. Both cotinine and, especially, nicotine were shown to be strongly concentrated in cervical mucus compared with serum levels. These findings confirm the association shown in epidemiologic studies of smoking and cervical neoplasia. It cannot be concluded that smoking is a genuine cause of cervical neoplasia. However, the results support the concept that smoking is a true risk factor in cervical neoplasia.

Cigarette smoking and plasma cholesterol

Plasma cholesterol levels were determined for 51,723 participants of community-based cholesterol screenings in 10 United States cities during 1988. Among white adult men and women under the age of 60 without other cardiovascular disease risk factors, a dose-response relationship was found between the number of cigarettes smoked per day and increasing levels of plasma cholesterol. In men aged 18 to 60 years, average plasma cholesterol increased by 0.33 mg/dl for each cigarette smoked (p less than 0.001); in women aged 31 to 50 years, average plasma cholesterol increased by 0.48 mg/dl for each cigarette smoked (p less than 0.001). Plasma cholesterol levels among ex-smokers were found to be similar to those of nonsmokers. No association between cigarette smoking and levels of plasma cholesterol was observed in men and women over age 60. Possible mechanisms for this observed relationship include an antiestrogenic effect of cigarette smoking that makes the observation more noticeable in younger female cohorts, enhanced lipolysis that increases levels of plasma free fatty acids, or differences in dietary intake between smokers and nonsmokers.

Tobacco sidestream smoke: Uptake by nonsmokers

Some epidemiological studies indicate an association between passive smoking and an increased risk for cancer, especially for cancer of the lung. Other reports, however, have failed to confirm these findings. Biochemical analyses of the physiological fluids for markers of exposure to tobacco smoke are needed as measurements of the uptake of smoke components by nonsmokers and for the estimation of relative cancer risk to passively exposed persons compared with that to active cigarette smokers. This communication reports the uptake of carbon monoxide, hydrogen cyanide, and nicotine after passive smoke exposure under controlled conditions. The results indicate that salivary nicotine values reflect the level of recent passive smoke exposure within an hour and that urinary cotinine values indicate the level of passive smoke exposure in the preceding hours. N-Nitrosoproline has been shown to serve as an indicator of endogenous N-nitrosamine formation in cigarette smokers: yet, preliminary studies do not indicate that urinary excretion of N-nitrosoproline is increased following short-term passive smoke exposure. In infants, first field studies suggest a correlation between exposure to tobacco-smoke-polluted environments and levels of cotinine in both serum and urine.

Biochemical validation of self-reported exposure to environmental tobacco smoke.

Biochemical validation of reported exposure to environmental tobacco smoke (ETS) lends credibility to epidemiological studies investigating the association of passive inhalation of smoke to respiratory disease or lung cancer. In the current study, a series of questions regarding ETS exposure was self-administered to nonsmokers and self-reported intensity of exposure was compared with cotinine levels in urine samples obtained on site. The target population of this study was a group of municipal workers who reported exposure in a domestic setting and/or in the workplace. When asked if they were exposed to ETS on social occasions, both males and females who responded positively had higher urinary cotinine levels (P less than 0.02) than those who gave a negative response. Mean urinary cotinine concentrations were found to be elevated in both men and women who reported that they lived with a smoker. Cotinine levels in the urine of those reporting exposure were over twice as high as those in the urine of respondents who denied having been exposed. ETS exposure in the home was the greatest contributor to increased urinary cotinine levels in both men and women. Among individuals who were exposed at work only, the reported degree of exposure agreed well with the mean urinary cotinine values. Those findings emphasize that the validation of exposure status with a biomarker is an essential prerequisite for epidemiological studies investigating passive smoking.

Population screening for plasma cholesterol: Community-based results from Connecticut

Plasma cholesterol levels were measured in 15,892 participants of a cholesterol screening in Hartford, Connecticut. The screenees were typically health conscious as evidenced by the low prevalence of current cigarette smoking (12%). However, more than 25% of the men and 35% of the women over the age of 50 years had cholesterol levels in excess of 240 mg/dl, placing them at moderate to high risk for coronary artery disease. Results showed a pronounced dose-response relationship between cigarette smoking and cholesterol levels in men of all age groups and in women of premenopausal age; however, average cholesterol levels were similar in exsmokers and subjects who had never smoked. Risk analysis revealed a strong positive association between cholesterol and prevalence of nonfatal myocardial infarctions. Notably patients with newly diagnosed hypercholesterolemia reduced their cholesterol levels an average of 40 mg/dl under the care of a physician. These results indicate that cholesterol screening coupled with physician follow-up and treatment can have a substantial impact in lowering cholesterol levels and the attendant risk of cardiovascular disease.

Cigarette smoking as a risk for cardiovascular disease. Part VI. Compensation with nicotine availability as a single variable

In a biochemical investigation of human smoking behavior with filter cigarettes with high draw resistance that varied only in nicotine yield, we attempted to determine which nicotine levels provide desired nicotine intake with a minimum of physiologic and biochemical consequences. Twelve prescreened subjects were divided into two study groups and supplied with cigarettes that varied in nicotine delivery. Both groups were initially monitored while smoking their usual cigarette. At the following visit, smokers in group 1 received incremental increases and smokers in group 2 received incremental decreases in nicotine levels in assigned cigarettes. All subjects were monitored upon first exposure, after 1 week of acclimatization to each experimental cigarette, and upon return to their usual brands. Subjects in both groups were unable to compensate fully for their nicotine uptake from the lowest nicotine cigarette. In subjects in group 1, new nicotine baselines began to develop after 1 week of acclimatization to cigarettes containing 0.9 and 1.3 mg nicotine. New baseline nicotine levels were also noted in subjects receiving decreases in nicotine (group 2) after smoking the cigarette containing 1.3 mg nicotine for 1 week. Carboxyhemoglobin concentrations did not differ from those measured after the usual‐brand cigarettes. Plasma cotinine concentrations increased as nicotine content per cigarette increased, except when subjects smoked a 1.3 mg nicotine cigarette. Plasma thiocyanate levels did not vary in either group. Systolic and diastolic blood pressures were generally not different from control values.

Thyroid hormone levels and cigarette smoking in baboons.

Abstract Using a primate animal model, two studies were undertaken to examine the effects of cigarette smoking on thyroid hormone levels. In study 1, mean total triiodothyronine (total T3) and mean total thyroxine (total T4) levels were measured in two groups of baboons (Papio cynocephalus) who were taught to smoke cigarettes using operant conditioning techniques. The smokers were divided into established and naive smokers according to pack-years of exposure. A control group of never-smoker baboons was included for comparison. Blood sampling was done after long-term cigarette consumption and again 1 week after cigarette deprivation. In the naive smoker group, mean total T3 concentrations were reduced below control group values (P < 0.05). After cigarette deprivation for 1 week, mean total T3 values returned to normal. No significant differences in total T4 levels were observed in either group. In study 2, we assessed some other indices of thyroid function. The same groups of baboons were divided into good and poor smokers by plasma cotinine and blood carboxyhemoglobin (% COHb) levels during 28 weeks of cigarette smoking activity. Immediate fluctuations and reductions in total T3 levels were observed that were not accompanied by reductions in total T4. The animals were then cigarette deprived for 1 week and blood samples were obtained every other day during this period. Significant increases in total T3 concentrations were observed in poor smokers immediately after cessation. Both groups also exhibited significant reductions (P < 0.05) in T3 uptake and free T4 index (FT4I) when compared to control group values. These data suggest that poor smokers are more susceptible to thyroid hormone level shifts than more established smokers, since the established smokers become habituated to the compounds contained in cigarette smoke through repeated exposure.

Cigarette smoking as a risk for cardiovascular disease III: Biochemical effects with higher nicotine yield cigarettes

Subjects who smoked a medium range nicotine yield cigarette were given a higher nicotine yield cigarette (an increase of 0.34 mg nicotine) to smoke ad libitum for two weeks. Plasma nicotine, cotinine, thiocyanate and blood carboxyhemoglobin levels were determined as well as various physiological parameters including heart rate and blood pressure. Increases in plasma nicotine were most directly correlated to heart rate when smokers were first challenged with a higher nicotine yield cigarette (r = 0.85); less directly correlated after a two-week acclimatization period (r = 0.42) and poorly related to their customary product (r = 0.23). Interestingly, it was noted that subjects did not compensate for higher nicotine yield by smoking fewer cigarettes per day when incremental nicotine changes were realistic. They did, however, show higher plasma nicotine, thiocyanate and an upward trend in plasma cotinine with the stronger cigarettes. These increases in cigarette constituents present in plasma, coupled with increasing correlation of heart rate and nicotine uptake, lead us to suggest that uptitration of smokers might cause them to establish new baseline levels. These findings have important health implications in light of recent suggestions to increase the nicotine yet decrease the tar of cigarettes in an attempt to overcome smoker compensation phenomena observed with low yield products.

Uptake of sidestream smoke by syrian golden hamsters

An inhalation bioassay with Syrian golden hamsters is being conducted to evaluate the toxic and carcinogenic potential of cigarette sidestream smoke (SS) relative to mainstream smoke (MS). A Hamburg II smoking machine is used to deliver MS by nose-only exposure to hamsters and a modification allows for the simultaneous collection of SS for whole-body delivery to a different rack of animals. The tolerated dose of SS was determined by varying the air/smoke dilutions drawn through the animal restrainers. Preliminary data indicated that 20% carboxyhemoglobin (COHb) could be obtained in SS-exposed animals without fatality. Optimum exposure levels were determined. Monthly measurements of COHb, nicotine and cotinine indicate that the SS-exposed animals are absorbing slightly higher amounts of these smoke constituents than the MS-exposed hamsters. Tumor incidence and carcinogenicity data are being collected through complete necropsy and histology protocols and uptake data continue to be collected. These studies should help elucidate the carcinogenic potential of SS which has been suggested from its composition and from recent epidemiological data of cancer incidence in non-smokers.