Simultaneous mainstream-sidestream smoke exposure systems II. The rat exposure system

A system for exposing rats to mainstream (MS) and sidestream (SS) smoke simultaneously from the same cigarette, and monitoring procedures, are described in detail. The equipment and procedures were used to expose Sprague-Dawley rats to mainstream smoke and to target deliveries of 10, 25, or 50% of the total SS smoke for 17 weeks. The estimated total particulate matter (TPM) dose was highly correlated with the increase in percent COHb for MS and SS smoke, but the COHb/TPM relationships were different for the 2 kinds of smoke. All SS TPM doses were much lower than the MS TPM dose, and the COHb/TPM ratio for SS smoke was much higher than for MS smoke. The TPM dose and per cent COHb for SS smoke were highly correlated with the per cent of SS sent to the exposure chambers. There were no significant differences in the total weight changes during the study for any of the smoke exposed groups, but weight changes during the 12-17-week period for sidestream exposed groups were inversely correlated with the level of sidestream exposure.     

Measurements of weight and activity in male mice following inhalation of cannabis smoke in a controlled smoke exposure chamber

A smoke exposure device delivering marijuana smoke to mice is described. Standardized smoking conditions were achieved with this unit yielding a concentration of 0.123 mg/liter of delta-9-tetrahydrocannabinol in air. A significant (p < 0.05) decrease in locomotor activity in treated animals was seen following the fourth treatment in a 21-day study where animals were exposed three times weekly for 3 weeks. A significant decrease in body weight for marijuana-exposed animals was also noted. In another study, animals were exposed daily for 7 days. Locomotor activity was significantly decreased in marijuana-exposed animals on Days 6 and 7. There was no significant change in body weight. Following removal from the exposure chamber, the marijuana-exposed animals showed transient hyperactivity (1–3 min) followed by a period of depressed activity lasting 1 hr. Some tolerance to the placebo smoke was seen after the fourth treatment in both studies. Cumulative effects were seen following repeated exposures. These preliminary data suggest that inhalation of marijuana smoke will initiate behavioral changes in mice.     

Pharmacokinetics of nicotine in rats after multiple-cigarette smoke exposure

The pharmacokinetics of nicotine and its major metabolites was evaluated in male rats after multiple-cigarette smoke exposure. A smoke-exposure apparatus was used to deliver cigarette smoke to the exposure chamber. The rats were exposed to smoke from a single cigarette every 8 hr for 14 days and to the smoke of a cigarette spiked with radiolabeled nicotine on the 15th day. Blood and urine samples were collected at timed intervals during the 10-min smoke-exposure period of the last cigarette and up to 48 hr thereafter. Nicotine, cotinine, and other polar metabolites were separated by thin-layer chromatography and quantified by liquid scintillation counting. The data were analyzed by computer fitting, and the derived pharmacokinetic parameters were compared to those observed after a single iv injection of nicotine and after a single-cigarette smoke exposure. The results indicated that the amount of nicotine absorbed from multiple-cigarette smoke was approximately 10-fold greater than that absorbed from a single cigarette. Also, unlike the single-cigarette smoke exposure experiment, nicotine plasma levels did not decay monotonically but increased after the 5th hr, and high plasma concentrations persisted for 30 hr. The rate and extent of the formation of cotinine, the major metabolite of nicotine, were decreased as compared with their values following a single-cigarette smoke exposure. It was concluded that nicotine or a constituent of tobacco smoke inhibits the formation of cotinine and may affect the biotransformation of other metabolites. Urinary excretion tended to support the conclusions that the pharmacokinetic parameters of nicotine and its metabolites were altered upon multiple as compared to single dose exposure.     

Effects of graded smoke inhalation on subsequent cigarette smoking behavior

Five smokers smoked a cigarette ad libitum one minute after inhaling either 0, 2, 4, 8 or 12 puffs of tobacco smoke according to a standardized smoking regimen. Heart rate and expired air carbon monoxide levels increased in a linear manner with increasing number of pretreatment puffs. Subjects took fewer puffs on, and spent less time smoking, and puffing on, the cigarette as the number of pretreatment puffs increased. The duration of individual puffs decreased with successive puffs as the cigarette was smoked, but was not affected by the puff pretreatments. Intervals between successive puffs (interpuff intervals) generally increased over the first half, and leveled off or decreased over the second half of the cigarette. Interpuff intervals occurring early in the cigarette tended to increase after the 12-puff pretreatment. The results are consistent with the suggestion that the observed increase in interpuff interval as a cigarette is smoked is the result of a satiation process.     

Studies on the deposition and distribution of catechol from whole cigarette smoke in BC3F1Cum mice

Tritiated catechol has been used to follow the pharmacokinetics and metabolic fate of inhaled catechol in cigarette smoke in $\text{BC3F1}\text{Cum}$ mice. The presence of [³H]catechol in the smoke was verified by silica gel chromatography, high-performance liquid chromatography, and gas chromatography/mass spectrometry. Mice were exposed to 10% ($\text{v}\text{v}$) 2R1 cigarette smoke on the Walton Horizontal Smoking Machine under standard conditions of 35 ml puff volume, 2 sec/puff, 10 puffs/cigarette. The deposition and distribution of inhaled catechol were determined in all internal tissues, urine, and feces. Data showed that clearance was occurring during the 10-min smoke exposure period. Immediately after exposure, over 50% of the radioactivity was found in the blood, with 10% found in the lung, and approximately 12% in the respiratory tract. Over 91% of the inhaled radioactivity was found in the urine 120 min after exposure. Less than 0.5% of the total dose was found in the lung at this time. We conclude that catechol in smoke is rapidly absorbed, redistributed, and excreted from mice exposed to whole cigarette smoke.     

Inhalation toxicity studies on cigarette smoke. V. Deposition of smoke particles in the respiratory system of rats under various exposure conditions

This paper describes a dosimetry experiment on rats which was designed to make a contribution towards the optimisation of exposure conditions for inhalation toxicology studies with smoke aerosols. The main conclusions drawn from the work are: (i) Under continuous exposure conditions the deposition of total particulate matter (TPM) in the respiratory system and carboxyhaemoglobin (COHb) levels in blood were linearly dependent on the concentration of smoke in the exposure chamber. (ii) Intermittent exposure gave relatively lower TPM deposition compared to continuous exposure, even after allowing for differences in actual exposure times. (iii) For arithmetically equivalent exposure levels, short exposure to high concentration gave greater TPM deposition than long exposures to low smoke concentrations. (iv) There was a good correlation between lower respiratory system (LRS) and lung deposition of TPM and blood COHb level for both continuous and intermittent exposure conditions. These findings are discussed in relation to the conduct of inhalation studies with tobacco smoke.     

Cigarette smoke inhalation and blood dose markers in beagle dogs

The time course of three tobacco-related blood dose markers was determined in beagle dogs during and after two-, four-, and six-cigarette acute smoke exposures. Venous blood samples were taken from a jugular catheter at selected intervals and analyzed for nicotine, cotinine, and carboxyhemoglobin (COHb). Smoke was generated from reference cigarettes (1.91 mg of nicotine) by a machine and delivered to the trachea of tracheostomized dogs. Cigarettes were chain-smoked in pairs with a 5-min break between pairs when four or six cigarettes were smoked. Blood nicotine, cotinine, and COHb levels all rose during smoke inhalation. Peak values of COHb were dosimetrically related to the number of cigarettes, while peak nicotine and cotinine values were not related consistently. The exposure protocol used in this study produced blood levels of all three measured parameters which were in the range of published values for human smokers, so that it is possible to approximate acute human cigarette smoke inhalation in the beagle dog model.     

Increased nasal epithelial ciliary beat frequency associated with lifestyle tobacco smoke exposure

The ciliated epithelium of the respiratory airways is one of the first vital systemic surfaces in contact with the ambient air. Ex vivo nasal epithelial ciliary beat frequency (CBF) at room temperature is on the order of 7–8 Hz but may be stimulated by irritant exposure. The upregulation of CBF in response to acute irritant exposure is generally considered to be a transient event with eventual return to baseline. However, studies of CBF dynamics in response to typical lifestyle exposures are limited. This study assessed nasal epithelial CBF among human subjects as a function of quantifiable lifestyle tobacco smoke exposure. Nasal epithelial biopsies were obtained from human subjects with well documented histories of tobacco smoke exposure. CBF was determined using a digital photometric technique and concurrent assays of nasal nitric oxide and urine cotinine and creatinine were performed. Mean CBF among active smokers and non-smokers exposed to environmental tobacco smoke (ETS) was elevated over non-smokers. Although there were dramatic differences in relative levels of tobacco smoke exposure, CBF values among tobacco smoke-exposed groups were comparable. Parallel in vitro studies of cultured nasal epithelium exposed to cigarette smoke condensate further supported these observations. These studies suggest that persistent elevation in nasal epithelial CBF is an early, subtle, physiologic effect associated with lifestyle tobacco smoke exposure. The molecular mechanisms that upregulate CBF may also create a cell molecular milieu capable of provoking the eventual emergence of more overt adverse health effects and the pathogenesis of chronic airway disease.     

The pharmacological activity of inhalation exposure to marijuana smoke in mice

Although the majority of cannabinoid users smoke marijuana, the preponderance of laboratory animal research is based on administration of Delta9-tetrahydrocannabinol (Delta9-THC) or other cannabinoid agents via injection. The aim of the present study was to evaluate the impact of inhaling marijuana, or ethanol-extracted placebo smoke in the mouse model of cannabinoid activity by assessing inhibition of spontaneous activity, antinociception, catalepsy, and body temperature. In order to determine dosimetry, blood levels of Delta9-THC were obtained following either marijuana exposure or intravenous injection of Delta(9)-THC. Inhalation exposure to marijuana produced dose-related increases in antinociception and catalepsy, with estimated ED50 doses of Delta9-THC of 2.4 and 3.8 mg/kg, respectively. However, hypothermia and locomotor depression occurred in both the placebo- and marijuana-exposed mice. The CB1 receptor antagonist, SR 141716A antagonized the antinociceptive effects of marijuana (AD50 = 0.6 mg/kg), but only slightly decreased marijuana-induced catalepsy, and failed to alter either the hypothermic or locomotor depressive effects. In contrast, SR 141716A antagonized the antinociceptive, cataleptic, and hypothermic effects of intravenously administered Delta9-THC in mice that were exposed to air alone, though all subjects exhibited locomotor depression, possibly related to the restraint. In accordance with reports of others, these data suggest that exposure to smoke alone has pharmacological consequences. Our findings also indicate that marijuana-induced antinociception is mediated through a CB1-receptor mechanism of action and are consistent with the notion that Delta9-THC is mainly responsible for this effect.     

Alterations in prostacyclin and thromboxane formation by chronic cigarette smoke exposure: temporal relationships and whole smoke vs. gas phase

Chronic cigarette smoke exposure in vivo causes decreased conversion of [14C]arachidonic acid (AA) to prostacyclin (PGI2) by isolated aortic tissue and increased conversion to thromboxane (TXA2) by isolated platelets from rats. Alterations in the PGI2/TXA2 balance may be part of the mechanism through which smoking increases the risk of cardiovascular disease. To study the influence of smoke exposure duration on this response, male rats were exposed daily to 10 puffs of freshly generated cigarette smoke. Animals were killed after 1, 4, 14, 28 and 57 days of smoke exposure and 3, 7, 14 and 28 days after cessation of the 57-day of smoke-exposure regimen. Elevated carboxyhemoglobin levels during the smoke-exposure sessions verified smoke (gas phase) inhalation. Statistically significant alterations in prostacyclin synthesis preceded those of thromboxane. A decrease of 20-25% (P less than 0.05) in PGI2 production from [14C]AA in isolated aortic tissue was found beginning 28 days after smoke was initiated and quickly rebounded when smoke exposure was terminated. Increased production of TXA2 from [14C]AA by isolated platelets became statistically significant (P less than 0.05) on the 57th day and returned to normal 7-14 days after cessation of smoke exposure. To determine the effect of gas phase constituents on the PGI2/TXA2 balance a second series of experiments divided male and female Sprague-Dawley rats into sham, whole smoke and gas phase groups. Gas phase was produced by passing whole smoke through a Cambridge filter to remove particulate matter. Per cent COHb averaged 1.4 for sham, 7.8 for whole smoke and 9.4 for gas phase groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pulmonary inflammation in mice exposed to mainstream cigarette smoke

Male C57Bl/6 (C57) and ICR mice were exposed by nose-only inhalation to mainstream cigarette smoke (MS) from 2R4F reference cigarettes, at concentrations of 75, 250, and 600 microg of total particulate matter (TPM) per liter, for up to 6 mo. Respiratory-tract tissue (nose, larynx, and lung), blood, and bronchoalveolar lavage fluid (BALF) samples were collected and analyzed at several time points. Blood samples were analyzed for biomarkers of exposure (COHb and nicotine). BALF was analyzed for biomarkers of cell injury, inflammation, oxidative stress, enzyme activity, and cytokines. Blood COHb and plasma nicotine concentrations increased in a dose-dependent manner, confirming smoke exposure. Mild emphysema was observed following 28 wk of exposure. Macrophage accumulation and inflammatory infiltrates were observed around the alveolar ducts and adjacent vasculature. There was an approximately 13% increase in mean linear intercept (Lm) only in ICR mice exposed to 600 microg/L TPM. There were no significant changes in biomarkers of oxidative stress secondary to smoke exposures; however, 8-isoprostane significantly increased following the 13-wk post-inhalation period. BALF macrophage and neutrophil counts were rapidly and consistently elevated, while lymphocyte counts gradually increased over time. MS-induced inflammatory responses observed in this study are comparable to changes reported in chronic smokers, supporting the role of chronic inflammation in the pathogenesis of emphysema. However, mild emphysema in minimal numbers of mice suggests that MS exposure concentration and/or duration in the current study were not sufficient to induce a definitive emphysema phenotype.     

Prior exposure to aged and diluted sidestream cigarette smoke impairs bronchiolar injury and repair.

The bronchiolar injury/repair response to naphthalene (NA) in mice includes acute distal airway epithelial injury that is followed by epithelial proliferation and redifferentiation, which result in repair of the epithelium within 14 days. To test whether prior exposure to aged and diluted sidestream cigarette smoke (TS) would alter the injury/repair response of the airway epithelium, adult mice were exposed to either filtered air (FA) or smoke for 5 days before injection with either corn oil carrier (CO) or naphthalene. Mice were killed 1 and 14 days after naphthalene injury. Lung and lobar bronchus were examined and measured using high-resolution epoxyresin sections. The control group (FACOFA) that was exposed to filtered air/corn oil/filtered air contained airway epithelium similar to untreated controls at all airway levels. The group exposed to tobacco smoke/corn oil/filtered air (TSCOFA) contained some rounded cells in the small airways and some expansion of the lateral intercellular space in the larger airways. Necrotic or vacuolated cells were not observed. As expected, the epithelium in the group exposed to filtered air/naphthalene/filtered air (FANAFA) contained many light-staining vacuolated Clara cells and squamated ciliated cells within distal bronchioles during the acute injury phase. Repair (including redifferentiation of epithelial cells and restoration of epithelial thickness) was nearly complete 14 days after injury. The extent of Clara cell injury, as assessed in lobar bronchi, was not different between the four groups. Although the FANAFA group contained greater initial injury in the distal airways at 1 day, the group exposed to tobacco smoke/naphthalene/filtered air (TSNAFA) had the least amount of epithelial repair at 14 days after naphthalene treatment; many terminal bronchioles contained abundant squamated undifferentiated epithelium. We conclude that tobacco smoke exposure prior to injury (1) does not change the target site or target cell type of naphthalene injury, since Clara cells in terminal bronchioles are still selectively injured; (2) results in slightly diminished acute injury from naphthalene in distal bronchioles; and (3) delays bronchiolar epithelial repair.     

Passive cannabis smoke exposure and oral fluid testing. II. Two studies of extreme cannabis smoke exposure in a motor vehicle

Two studies were conducted to determine if extreme passive exposure to cannabis smoke in a motor vehicle would produce positive results for delta-tetrahydrocannabinol (THC) in oral fluid. Passive exposure to cannabis smoke in an unventilated room has been shown to produce a transient appearance of THC in oral fluid for up to 30 min. However, it is well known that such factors as room size and extent of smoke exposure can affect results. Questions have also been raised concerning the effects of tobacco when mixed with marijuana and THC content. We conducted two passive cannabis studies under severe passive smoke exposure conditions in an unventilated eight-passenger van. Four passive subjects sat alongside four active cannabis smokers who each smoked a single cannabis cigarette containing either 5.4%, 39.5 mg THC (Study 1) or 10.4%, 83.2 mg THC (Study 2). The cigarettes in Study 1 contained tobacco mixed with cannabis; cigarettes in Study 2 contained only cannabis. Oral fluid specimens were collected from passive and active subjects with the Intercept Oral Specimen Collection Device for 1 h after smoking cessation while inside the van (Study 1) and up to 72 h (passive) or 8 h (active) outside the van. Additionally in Study 1, Intercept collectors were exposed to smoke in the van to assess environmental contamination during collection procedures. For Study 2, all oral fluid collections were outside the van following smoking cessation to minimize environmental contamination. Oral samples were analyzed with the Cannabinoids Intercept MICRO-PLATE EIA and quantitatively by gas chromatography-tandem mass spectrometry (GC-MS-MS). THC concentrations were adjusted for dilution (x 3). The screening and confirmation cutoff concentrations for THC in neat oral fluid were 3 ng/mL and 1.5 ng/mL, respectively. The limits of detection (LOD) and quantitation (LOQ) for THC in the GC-MS-MS assay were 0.3 and 0.75 ng/mL, respectively. Urine specimens were collected, screened (EMIT, 50 ng/mL cutoff), and analyzed by GC-MS-MS for THCCOOH (LOD/LOQ = 1.0 ng/mL). Peak oral fluid THC concentrations in passive subjects recorded at the end of cannabis smoke exposure were up to 7.5 ng/mL (Study 1) and 1.2 ng/mL (Study 2). Thereafter, THC concentrations quickly declined to negative levels within 30-45 min in Study 1. It was found that environmentally exposed Collectors contained 3-14 ng/mL in Study 1. When potential contamination during collection was eliminated in Study 2, all passive subjects were negative at screening/confirmation cutoff concentrations throughout the study. Oral fluid specimens from active smokers had peak concentrations of THC approximately 100-fold greater than passive subjects in both studies. Positive oral fluid results were observed for active smokers 0-8 h. Urine analysis confirmed oral fluid results. These studies clarify earlier findings on the effects of passive cannabis smoke on oral fluid results. Oral fluid specimens collected in the presence of cannabis smoke appear to have been contaminated, thereby falsely elevating THC concentrations in oral fluid. The risk of a positive test for THC was virtually eliminated when specimens were collected in the absence of THC smoke.     

Free lung cell response of mice and rats to mainstream cigarette smoke exposure

Male C57BL mice and F-344 rats were exposed through nose only to fresh mainstream smoke from one University of Kentucky Reference cigarette (2R1) daily under standardized conditions. At different exposure points, the lungs of room control (RM), sham control (SH), and smoke-exposed (SM) animals were lavaged and the number, composition, and properties of bronchoalveolar lavage (BAL) cells were studied. Significantly elevated levels of blood COHb and pulmonary aryl hydrocarbon hydroxylase activity indicated effective inhalation of smoke by animals. The BAL cell analysis showed that cigarette smoke induced a five- to sevenfold increase in the number of BAL cells in mice following 10- to 12-week exposure. The proportion of neutrophils (PMN) increased to about 18 +/- 3% in SM mice as compared to less than 1% in controls. Cessation of smoke treatments returned the PMN levels to those of controls within 5 weeks. Unlike mice, smoke exposure for up to 32 weeks failed to induce appreciable changes in the number and proportion of macrophages and neutrophils in rats. Large brown macrophages were observed in SM groups of both species. Functional analysis demonstrated that the BAL cells from SM mice but not rats released greater amounts of superoxides than controls under resting and phagocytically stimulated conditions. Enzymatic analysis of macrophages showed that the activity of N-acetylglucosaminidase was increased in SM groups of both species. The activity of 5'nucleotidase was significantly reduced in macrophages from SM mice but not rats. Activity of leucine aminopeptidase remained unaltered in both species. These results demonstrate distinct differences in the response of mice and rats to identically generated cigarette smoke.     

Deposition and distribution of the total particulate matter of cigarette smoke in mice using a large-capacity smoke exposure system

A newly developed automatic smoke exposure machine (SEM II) was used to generate [¹⁴C]dotriacontane-labeled University of Kentucky reference 2A1 or 2R1 cigarette smoke. The SEM II is a large-capacity (480 mice) dynamic smoke exposure system in which smoke is routed through the animal containment system as a continuously flowing stream. Mice are restrained about the neck in stock-like holders for “nose-only” exposure. Using standard smoke exposure conditions, the deposition and internal distribution of the total particulate matter (TPM) from cigarette smoke was determined in BC3Fl/Cum male and female mice. Results show: (a) smoke exposure conditions can be varied so that deposition from 30 to 200 μg TPM/lung can be obtained, (b) 80–90% of the TPM deposition was found in the respiratory tissues, (c) the mouse-to-mouse variation for TPM deposition in pulmonary tissue was ～20%, (d) similar deposition and distribution of TPM was observed in male and female mice, and (e) deposition and distribution of TPM was not altered in mice exposed to smoke on a daily basis over a 6-month period of time.     

Protective antioxidant mechanisms in rat and guinea pig tissues challenged by acute exposure to cigarette smoke.

Cellular damage from reactive intermediates formed during xenobiotic biotransformation is prevented by the presence of adequate levels of antioxidant chemicals in the tissues. Equally important for cell protection is the rate at which these chemicals are replaced if tissue stores are depleted. The present experiments, using adult male Sprague-Dawley rats and Hartley guinea pigs, were conducted to ascertain what effects mainstream (MS) and sidestream (SS) tobacco smoke would have on the water-soluble, cytoplasmic antioxidants, ascorbic acid (AA) and reduced glutathione (GSH). The animals were exposed by nose-only inhalation to varying doses (40, 120, 240 puffs) of a 1:5 dilution of a 35-ml volume of freshly generated MS from cigarettes made from different types of tobacco and delivered by a B.-A.T-Mason inhalation apparatus. The animals were euthanized either immediately following exposure or at 3 and 6 h. The blood, lungs, liver, kidneys, heart and bladder were removed for the quantitation of AA and GSH following homogenization and deproteinization. Immediately following exposure to MS, dose-dependent decreases in pulmonary and renal GSH were observed in rats whereas, in guinea pigs, reductions in pulmonary, hepatic and renal GSH were observed only at the highest level of exposure. No reductions in tissue AA were observed in either species at any exposure level. In both species, blood levels of GSH and AA remained unchanged following exposure. Mainstream smoke (240 puffs) from flue-cured or dark, air-cured tobaccos elicited a significant, immediate reduction in pulmonary and renal GSH, but MS from low tar, filter cigarettes was without effect. Within 3 h of exposure, GSH in all tissues has returned to pre-exposure levels. Whole-body, chamber exposure to concentrated SS, generated from smouldering cigarettes, caused a dose-dependent reduction in rat pulmonary, hepatic, renal, cardiac and bladder muscle GSH but only affected pulmonary GSH in the guinea pig. Lesser effects were observed in tissues of rats exposed to diluted SS. In the rat, a comparison of the results of diethylmaleate- and smoke-induced depletion of tissue GSH suggested that, even at exceptionally high levels of exposure, there was a significant store of GSH in tissues that did not interact with tobacco smoke.     

Breathhold duration and response to marijuana smoke

Marijuana smokers are frequently observed to hold the smoke in their lungs for prolonged periods (10-15 sec) apparently in the belief that prolonged breathholding intensifies the effects of the drug. The actual influence of breathhold duration on response to marijuana smoke has not been studied. The present study examined the effects of systematic manipulation of breathhold duration on the physiological, cognitive and subjective response to marijuana smoke in a group of eight regular marijuana smokers. Subjects were exposed to each of three breathhold duration conditions (0, 10 and 20 sec) on three occasions, scheduled according to a randomized block design. A controlled smoking procedure was used in which the number of puffs, puff volume and postpuff inhalation volume were held constant. Expired air carbon monoxide levels were measured before and after smoking to monitor smoke intake. Typical marijuana effects (increased heart rate, increased ratings of "high" and impaired memory performance) were observed under each of the breathhold conditions, but there was little evidence that response to marijuana was a function of breathhold duration.     

Comparative 13-week inhalation study of cigarette smoke from cigarettes containing cast sheet tobacco

A subchronic, nose-only inhalation study was conducted to compare the effects of mainstream smoke from a reference cigarette containing conventional reconstituted tobacco sheet at 30% of the finished blend to mainstream smoke from cigarettes containing 10% or 15% cast sheet (a specific type of reconstituted tobacco sheet) substituted for part of the conventional reconstituted tobacco. Male and female Sprague-Dawley rats were exposed for 1 h/day, 5 d/wk, for 13 wk to mainstream smoke at 0, 0.06, 0.20, or 0.80 mg wet total particulate matter per liter of air. Clinical signs, body and organ weights, clinical chemistry, hematology, carboxyhemoglobin (COHb), serum nicotine, plethysmography, gross pathology, and histopathology were determined. Exposure to cigarette smoke induced a number of changes in respiratory physiology, histopathology, and serum nicotine and COHb levels when compared to sham animals. When corresponding dose groups of reference and cast sheet mainstream smokes were compared, no biological differences were noted. At the end of the exposure period, subsets of rats from each group were maintained without smoke exposures for an additional 13 wk (recovery period). At the end of the recovery period, there were no statistically significant differences in histopathological findings observed between the reference and either cast sheet cigarette. Substitution of 10% or 15% cast sheet tobacco for conventional reconstituted tobacco sheet does not alter the inhalation toxicology of the mainstream smoke when compared to mainstream smoke from a reference cigarette containing conventional reconstituted tobacco sheet.     

Induction of aryl hydrocarbon hydroxylase in rat lung by marijuana smoke.

Rats were exposed to the smoke produced by burning four cigarettes of marijuana or of marijuana placebo material. Six hours later, maximally stimulated aryl hydrocarbon hydroxylase (AHH) activity was observed in the lung. Marijuana placebo material also induced pulmonary AHH activity. Actinomycin D (1 mg/kg) and cycloheximide (2 mg/kg), given ip 1 hr before exposure to marijuana smoke, partially blocked AHH induction. If rats were exposed repeatedly to marijuana smoke, they showed higher activities of pulmonary AHH within 6 hr of the last exposure than did animals exposed for the first time. Inhalation of marijuana smoke increased the number and size of debris-filled vacuoles in alveolar macrophages and in the ciliated cells of the bronchiolar epithelium. Smoke condensate from the marijuana material used in these studies contained 0.45 ng/mg of benzo(a)pyrene.     

Tobacco smoke exposure induces nicotine dependence in rats

Rationale

Tobacco smoke contains nicotine and many other compounds that act in concert on the brain reward system. Therefore, animal models are needed that allow the investigation of chronic exposure to the full spectrum of tobacco smoke constituents.

Objectives

The aim of these studies was to investigate if exposure to tobacco smoke leads to nicotine dependence in rats.

Methods

The intracranial self-stimulation procedure was used to assess the negative affective aspects of nicotine withdrawal. Somatic signs were recorded from a checklist of nicotine abstinence signs. Nicotine self-administration sessions were conducted to investigate if tobacco smoke exposure affects the motivation to self-administer nicotine. Nicotinic receptor autoradiography was used to investigate if exposure to tobacco smoke affects central α7 nicotinic acetylcholine receptor (nAChR) and non-α7 nAChR levels (primarily α4β2 nAChRs).

Results

The nAChR antagonist mecamylamine dose-dependently elevated the brain reward thresholds of the rats exposed to tobacco smoke and did not affect the brain reward thresholds of the untreated control rats. Furthermore, mecamylamine induced more somatic withdrawal signs in the smoke-exposed rats than in the control rats. Nicotine self-administration was decreased 1 day after the last tobacco smoke exposure sessions and was returned to control levels 5 days later. Tobacco smoke exposure increased the α7 nAChR density in the CA2/3 area and the stratum oriens and increased the non-α7 nAChR density in the dentate gyrus.

Conclusion

Tobacco smoke exposure leads to nicotine dependence as indicated by precipitated affective and somatic withdrawal signs and induces an upregulation of nAChRs in the hippocampus.