Effects Of Smokeless Tobacco Research Articles

Effect of short-term use of oral smokeless tobacco on random-pattern skin flap survival in rats

We investigated the effects of smokeless tobacco on the survival of random-pattern skin flaps in rats. Twenty rats were divided into two groups (n=10 each). In the experimental group 200 mg smokeless tobacco (Maras powder) (1 mg nicotine) was inserted intraorally once a day for seven days under general anaesthesia. It was not given to the control group, but the rats were similarly anaesthetised with ketamine. On day 8, plasma cotinine concentrations were measured. The random-pattern dorsal skin flaps measuring 3 x 10 cm were then raised and resutured. Percentage survival area was assessed after a further eight days. The mean (SD) survival was 39 (7)% in the experimental, and 65 (8.9)% in the control, groups (p=0.0001). The mean (SD) plasma cotinine concentrations were 124.4 (73) ng/ml and <10 ng/ml, respectively. Smokeless tobacco use increased the incidence of flap necrosis in random-pattern skin flaps in rats.

Smokeless Tobacco Extracts Activate Complementin Vitro:A Potential Pathogenic Mechanism for Initiating Inflammation of the Oral Mucosa

The use of smokeless tobacco has been linked to an increased incidence of inflammation of the buccal and gingival mucosa. However, the mechanisms by which smokeless tobacco initiates inflammation are not well understood. The complement cascade is a ubiquitous source of proinflammatory molecules and can be activated rapidly by a wide variety of agents. Therefore, the effect of smokeless tobacco on complement was investigated as a potential pathogenic mechanism for triggering inflammation of the oral mucosa. Aqueous extracts of loose leaf chewing tobacco (1S1), dry snuff (1S2), and moist snuff (1S3), added to normal human serum, depleted complement hemolytic activity in a dose-dependent manner. Experiments utilizing sera deficient in one specific complement component indicated that the smokeless tobacco-induced depletion of hemolytic activity was due largely to consumption of C3. Furthermore, assays designed to test the activity of the alternative pathway of complement clearly showed that all three extracts depleted the hemolytic activity of this pathway. Finally, all three smokeless tobacco extracts activated the alternative pathway since significantly elevated levels of the cleavage fragments iC3b and Bb were detected in extract-treated serum. High quantities of the classical pathway cleavage fragment C4d also were detected in serum treated with moist snuff (1S3). The results clearly demonstrate that smokeless tobacco extracts activate the alternative pathway and also suggest some measure of classical pathway activation. Activation of complement by smokeless tobacco may be a mechanism for initiating inflammation of the oral mucosa.

Leukoplakia

Leukoplakia as a medical term has been encumbered by a hodgepodge of synonyms and interpretations. It also has been a term associated with strong emotions with respect to an implied malignant potential. It now is regarded as potentially malignant, with varying degrees of risk for transformation. Leukoplakia at its major anatomic site of occurrence, the oral cavity, recently has been redefined, classified, and staged as a preamble to a more rational approach to the clinicopathologic study of leukoplakia. These changes are presented, as are current assessments of proliferative verrucous leukoplakia, verrucous hyperplasia, palatal white lesions, effects of smokeless tobacco on the oral mucosa, hairy leukoplakia, and lichen planus.

Inhibition of Neutrophil and Monocyte Defensive Functions by Nicotine

To learn more about the effects of smokeless tobacco on the defensive functions of neutrophils, we studied the influence of nicotine on these cells in vitro, looking at their bactericidal activity against oral pathogens, and at their ability to produce microbicidal reactive oxygen species (oxygen radicals). Exposure of human blood neutrophils to nicotine (0.01% to 0.1%) inhibited their ability to kill Actinomyces naeslundii, Actinobacillus actinomycetemcomitans, and Fusobacterium nucleatum. Although these concentrations of nicotine are high, such concentrations are relevant to phagocytes in the gingival sulcus, because smokeless tobacco contains 0.5% to 3.5% nicotine by dry weight. Nicotine had no such inhibitory effect when the killing assay was performed in an anaerobic environment, implying that nicotine preferentially affected oxygen-dependent killing mechanisms. To further investigate the effects of nicotine on production of oxygen radicals, neutrophils were primed with lipopolysaccharide and triggered with f-met-leu-phe or phorbol ester in the presence of nicotine. Nicotine inhibited production of superoxide anion (measured by reduction of cytochrome c) and hydrogen peroxide (measured by oxidation of phenol red). Nicotine inhibition of superoxide production was reversible by washing away the nicotine. By observing that nicotine inhibited the reduction of cytochrome c by reagent potassium superoxide, we determined that nicotine directly absorbed superoxide. In addition, by examining nicotine inhibition of the uptake of oxygen by neutrophils, we determined that nicotine also interfered with the production of oxygen radicals by these cells. Nicotine also inhibited production of superoxide and interleukin-1 beta by monocytes. Nicotine did not affect the viability of neutrophils and monocytes, as determined by their ability to exclude trypan blue dye. Inhibition of the aerobic antimicrobial functions of neutrophils and monocytes by nicotine may alter the microbial ecology of the oral cavity, and this might be one mechanism by which nicotine compromises the oral health of users of tobacco products.

Snuffing it Out: A Smokeless Tobacco Intervention with Athletes at a Small Private College

This study assessed attitude, knowledge, and use of smokeless tobacco (moist snuff, dry snuff, or chewing tobacco) among male varsity athletes at a small private college. Forty-three athletes were surveyed and 17 (40%) were found to be current users. Of these users, 27% reported dipping more than once a day, everyday; 25% of all users had noticed physical changes, such as white spots, inside their mouths. The wide acceptance of using smokeless tobacco was clear from the finding that 49% of those surveyed believed that athletes should be allowed to use "chew" during practice. These athletes also showed limited knowledge of the nicotine content and physiological effects of smokeless tobacco. As a result of the study, the authors developed a two-part intervention that included an educational presentation and oral screening by a local dentist and an administrative directive enforcing the National Collegiate Athletic Association's policy on smokeless tobacco use. A follow-up survey indicated the intervention had had some positive effects on the athletes' attitudes toward and knowledge about using smokeless tobacco. The survey also showed an increase from 4 to 7 in the number of former users who said they had stopped using smokeless tobacco.

Effects of smokeless tobacco on chemically transformed hamster oral keratinocytes: role of angiotensin I-converting enzyme.

The purpose of this study was to determine whether exposure of chemically transformed golden Syrian hamster oral epidermoid carcinoma cell (HCPC-1) cultures to smokeless tobacco extract (STE) is associated with a decrease in specific angiotensin I-converting enzyme (ACE) activity and whether this decrease potentiates bradykinin-induced cell growth. We found that STE induced a significant concentration- and time-dependent decrease in ACE activity in cultured HCPC-1 cells (P < 0.05). STE alone had no significant effect on cell number. Bradykinin alone induced a slight, but significant, increase in cell number (P < 0.05). These effects were significantly potentiated by STE (P < 0.01). We conclude that STE potentiates bradykinin-induced HCPC-1 cell growth, in part by attenuating specific ACE activity in these cells.

The effects of smokeless tobacco (oral snuff) on dynamic visual acuity

The effects of the use of smokeless tobacco (oral snuff) on observers’ ability to resolve moving targets were assessed under four levels of target velocity for both light and heavy tobacco users among college students. Each observer was tested in several sessions, half immediately following the use of tobacco and half with no preceding tobacco use. No differences were found across the two types of sessions for any of the velocity conditions. The results are discussed in terms of the practical consequences of nicotine nystagmus reported by previous investigators.

Immunostimulatory potential of smokeless tobacco extract in in vitro cultures of murine lymphoid tissues.

Despite numerous studies on the general toxicologic effects of smokeless tobacco (ST) little immunotoxicologic information is available. As a first step in assessing the potential activity of ST on the immune system, the effects of an aqueous extract of ST was studied in in vitro cultures of mouse lymphoid cells. There was a significant increase in the proliferation of spleen cells cultured with different concentrations of ST extract. The polyclonal IgM antibody responses as determined by protein A plaque assay were also elevated in ST stimulated spleen cell cultures. Similar immunostimulatory results were seen in the mesenteric lymph node cell cultures also. ST extract was able to stimulate the spleen cells of the immune defective CBA/N mice. The mitogenic ability of ST extract may not be due to lipopolysaccharide (LPS) contamination as determined by its response in the LPS resistant C3H/HeJ mice spleen cells. ST extract was mitogenic not only to B cells but also to T cells. However the magnitude of response was less in T cells than in B cells. The proliferation of T cells was not accompanied by secretion of IL-2 or expression of IL-2 receptors on T cells. However there was an increase of IL-1 activity in spleen cells cultured with ST extract. Finally, activation of B or T lymphocytes by ST did not result in the elevation of intracellular calcium levels. Since ST is consumed orally, the chronic immunostimulation by ST in oral mucosal lymphoid tissues may be associated with the increased incidence of gingivitis, leukoplakia and oral cancer seen in human ST users.

Effect of Smokeless Tobacco on Cardiovascular Risk Factors in Professional Baseball Players

Effect of Smokeless Tobacco on Cardiovascular Risk Factors in Professional Baseball Players

The effects of smokeless tobacco on performance and psychophysiological response

The effects of smokeless tobacco (ST) on psychophysiological response and performance were examined in two studies. Study 1 compared heavy and moderate ST users on psychomotor tasks (simple reaction time/anticipation time and choice reaction time/movement time) in ST (2 g amount) and no smokeless tobacco (NST) conditions. Moderate users performed significantly better than heavy users; there were no significant psychomotor performance differences comparing ST/NST conditions. Heart rate and blood pressure were elevated equally for both groups with the use of ST. Heavy users reported significantly less state anxiety in both ST and NST conditions than moderate users. Study 2 replicated the psychomotor performance results of study 1 when comparing nonusers and ST users who were given four dose amounts of ST (NST, 1/3 mean dose, mean dose, and 5/3 mean) over 4 d. For the math and Stroop tasks, ST use resulted in 12.1% and 10.4% better performance than the NST condition. Physiological measures showed significant elevations during the ST conditions. No differences between dose amounts of ST were apparent for any of the measures. These studies demonstrate that ST users perform better than nonusers when having to react to cognitively challenging task situations.

Effect of smokeless tobacco and tobacco-related chemical carcinogens on survival of ultraviolet light-inactivated herpes simplex virus

Low doses of ultraviolet (UV) light, x-rays, photodynamic treatment, or aflatoxins increase the survival of UV-irradiated virus in cells. This effect is postulated to occur by enhancement of the error-prone cellular repair function, which could also be associated with oncogenic cell transformation. The present study was designed to investigate whether treatment of green monkey kidney cells with water extract of snuff (snuff extract), benzo [a]pyrene, nicotine, or tobacco-specific N′-nitrosamines would result in enhanced survival of UV-irradiated herpes simplex virus (HSV). Exposure of the cells with snuff extract, benzo [a]pyrene, N′-nitrosonornicotine, or 4-( N-methyl- N′-nitrosamino)-1-(3-pyridyl)-1-butanone resulted in an enhancement of survival of UV-irradiated HSV type 1 compared with the control whereas exposure of the cells with nicotine did not. These data indicate that the water-extractable component of snuff and tobacco-related chemical carcinogens increase the cellular repair mechanism and provides for increased survival of UV-irradiated HSV.

Effect of smokeless tobacco use in humans on mucosal immune factors

To assess the effects of smokeless tobacco on the secretory immune system and dental caries, we examined users of smokeless tobacco and non-tobacco users. There were no significant differences in the prevalence of DMFS between users and non-users. There was significantly more salivary IgA, IgA2 and J-chain in users. Levels of salivary lysozyme and lactoferrin were significantly lower in users than controls. Because there was no difference in levels of secretory component in relation to the increased IgA levels of smokeless tobacco users, this suggests an effect of smokeless tobacco on secretory epithelial cells responsible for synthesis of secretory component, lysozyme and lactoferrin, and for the packaging of secretory component on IgA. There were only slight differences in salivary or serum antibody levels to Streptococcus mutans. These findings indicate that although smokeless tobacco has a significant influence on the synthesis of secretory IgA, the numbers of DMFS were similar between smokeless tobacco users and controls.

Effects of smokeless tobacco on murine splenic lymphocytes in in vitro cultures

Effects of smokeless tobacco on murine splenic lymphocytes in in vitro cultures

Effects of smokeless tobacco on the ability of secretory component to bind to the IgA/J chain complex

Previously, we reported that smokeless tobacco users have significantly higher levels of immunoglobulin A and J chain in whole saliva than non-tobacco users. Because there was no difference in levels of secretory component between the two groups, the proportion of secretory component/immunoglobulin A was significantly lower in users than non-users. There was no significant difference in antibody function. In the present study, we examined immunoglobulin A from whole saliva of users and non-users to determine the effect of smokeless tobacco on the ability of secretory component to bind to immunoglobulin A containing J chain. Whole saliva was passed over an affinity chromatography filter unit coupled with anti-alpha heavy chain-specific antibody followed by passage over a molecular sieve high-performance liquid chromatography column. Peaks were collected and examined for immunoglobulin A, J chain and secretory component by enzyme-linked immunosorbent assay. Saliva from users had three significantly larger peaks (3-4 fold) at 280 nm than non-users, confirming the presence of a higher concentration of immunoglobulin A in users. The secretory component/J chain and secretory component/immunoglobulin A ratios for the largest peak were significantly less in users. This indicates that smokeless tobacco has an effect on the ability of secretory component to bind to immunoglobulin A without a loss in antibody function. This may occur either prior to immunoglobulin A/J chain binding to secretory component receptors on secretory epithelial cells or internal to the epithelial cells. These studies provide further evidence for the role of secretory component in mucosal immunity.

Smokeless Tobacco Use and Health Effects Among Baseball Players

The effects of smokeless tobacco (ST) use were studied in 1109 members of major and minor league professional baseball teams during spring training in 1988. The prevalence of current ST use was 39%. The median age at initiation among users was 18 years, and the median duration of use was 5 years. Among users, 75% cited a snuff brand as their usual ST product. Oral leukoplakia was present in 46% (196/423) of current-week ST users and 1.4% (7/493) of nonusers (odds ratio, 60; 95% confidence interval, 28 to 130). Prevalence of oral leukoplakia among ST users increased with hours used per day and decreased with time lapsed since last use, and was higher in snuff users than in chewing tobacco users. Of the subjects with oral leukoplakia who underwent punch biopsy, 91 had benign hyperkeratosis and one had mild dysplasia. Overall prevalence of dental caries, gingivitis, and plaque did not differ between ST users and nonusers. In analyses confined to facial surfaces of mandibular incisor teeth, where ST is most commonly used, there were significant increases among users in both gingival recession and attachment loss. Users of ST did not differ from nonusers in blood pressure, pulse, total or high-density lipoprotein cholesterol level, or white blood cell count, but among users high-density lipoprotein cholesterol levels were inversely associated with serum cotinine levels. The major health effects of ST use among professional baseball players are oral leukoplakia and localized periodontal disease. The study population was young, physically fit, and characterized by relatively moderate short-term ST use.

Effect of smokeless tobacco on the development of the CD-1 mouse fetus.

The objective of this study was to examine the effect of an aqueous extract of smokeless tobacco (ST) on the development of the CD-1 mouse fetus. Three ST dosages were administered three times daily by gastric intubation during gestational days 1-17: 1 X ST equivalent to a dose of 4 mg nicotine/kg body weight, 3 X ST equivalent to 12 mg nicotine, and 5 X ST equivalent to 20 mg nicotine/kg body weight. Maternal plasma nicotine levels were determined 30 minutes after the second daily intubation at five different times during the gestational period. At these ST dosages, the weight gain of ST-treated dams was not significantly affected in comparison to treated controls, though the difference was significant (P less than .05) in comparison to untreated controls. The mean maternal plasma nicotine level for the low dosage (1 X) group was 99.0 ng/ml, which reasonably approximates human consumption levels. The 3 X ST and 5 X ST dosages produced higher nicotine plasma values, 398 ng/ml and 623 ng/ml, respectively, were considerably more toxic to the dams, and resulted in 18% and 31% maternal deaths. Fetal weights were reduced by 7.4% (P less than .001) in the highest ST dosage group (5 X), whereas at the 1 X and 3 X dosages fetal weight differences were not significantly different from treated controls. Resorptions increased in a dose-related manner (P less than .05), ranging from 4.7% in the 1 X, to 6.4% in the 3 X and 8.9% in the 5 X dosage compared to 3.2% in treated controls. External malformations were few and minor in extent. Internal malformations increased in a linear, dose-related manner (P less than .05). Placental weights were unaffected by ST. The results of skeletal examinations were inconclusive. Precocious ossification was seen in 60% and 70% of the parameters measured in the 1 X and 3 X dosage groups, respectively, in comparison to controls. In the 5 X ST group ossification levels were less than in controls for 30% of the parameters measured. Under these experimental conditions the lowest ST dosage (1 X) produced a negligible effect on the CD-1 mouse fetus and the dam. The highest ST dose (5 X) demonstrated embryotoxicity, growth retardation, few malformations, and maternal toxicity. The intermediate dose (3 X) showed a range of effects between the highest and lowest doses to both the fetus and the dam.

Effects of smokeless tobacco on the buccal mucosa of HMT rats.

Fifteen male and 15 female HMT rats, 6 months of age, received weekly applications of smokeless tobacco to the buccal mucosa for one year. Hyperorthokeratosis, acanthosis, numerous binucleate spinous cells, and subepithelial connective tissue hyalinization were observed, whereas verrucous carcinoma and squamous cell carcinoma were not seen. Karyotyping revealed that lymphocytes of tobacco-treated as well as control rats had normal chromosome number (2N = 42) and morphology. However, approximately 25% of buccal epithelial cells of the tobacco-treated rats were tetraploid and 5% octaploid, compared to only 11% tetraploid and no octaploid in the controls. Results indicate that the effects of smokeless tobacco on the rat buccal mucosa are similar to those observed in humans and that the mitotic process can be disturbed by tobacco treatment.

The acute effects of smokeless tobacco on transport and barrier function of buccal mucosa.

The acute effects of smokeless tobacco (ST) on buccal mucosal transport and barrier function were studied by means of in vivo and in vitro techniques. In humans, in vivo exposure to 0.5 g ST transiently increased the transmural electrical potential difference (PD). However, despite continued exposure, PD returned to baseline within 20 min. The mechanisms for these changes were explored by use of dog buccal mucosa mounted in Ussing chambers. Luminal exposure to a Ringer-extract of ST (EOST) increased PD and short-circuit current (Isc) and decreased electrical resistance (R), with changes reversible upon removal of EOST from the bath. Further, radioisotopic fluxes showed that the increase in Isc in EOST-exposed tissues was accompanied by increased absorption of electrolytes (Na, Cl, and other ions), and the decrease in R was accompanied by increased permeability to mannitol. Light microscopy of tissues exposed to EOST showed no morphological changes after exposure to 0.5 g of ST, but after exposure to 1.5-2.5 g of ST, dilated intercellular spaces were identified. Contact of aqueous media with ST led to the release of electrolytes and other soluble compounds into solution. To determine the effect of electrolyte release on buccal function, we exposed mucosae luminally to a solution with ion composition and/or osmolality similar to EOST or to one with an EOST previously dialyzed against Ringer. Solutions with similar ion composition and/or osmolarity changed PD, Isc, and R in a manner similar to EOST, while dialyzed-EOST had no effect. In addition, luminal nicotine produced effects different from EOST, decreasing PD and Isc and increasing R.(ABSTRACT TRUNCATED AT 250 WORDS)

Smokeless tobacco contains a nonnicotine inhibitor of bone metabolism

The effects of smokeless tobacco on bone were investigated using tibiae from chick embryos. The bones were cultured in nicotine (15–1500 μg/ml) or in smokeless tobacco extract (STE, 15 mg tobacco/ml culture medium) and the effects on bone glucose metabolism (oxygen consumption and lactate production) and collagen synthesis ([ 3H]proline hydroxylation) were tested in vitro. Only the highest concentration of nicotine tested produced substantial effects as demonstrated by decreased oxygen consumption and [ 3H]hydroxyproline content (82 and 90%, respectively) and by a 15% increase in lactate production. Results with STE, which contained nicotine concentrations ranging from 104 to 125 μg/ml, showed similarly reduced [ 3H]hydroxyproline content (90%), but oxygen consumption was only reduced 36%, while lactate production was elevated 60%. The ability of the bones to recover from treatment with STE was demonstrated by increased oxygen consumption and [ 3H]hydroxyproline content (18 and 78%, respectively) and decreased lactate production (47%) compared to the STE-treated bones. These findings suggest the following conclusions: (1) both nicotine and STE at concentrations found in the saliva of smokeless tobacco users stimulate glycolysis and markedly inhibit bone collagen synthesis and mitochondrial activity; (2) effects of STE on bone are not due to nicotine; and (3) under the conditions studied, bone partially recovers from the effects of STE.

Introduction: Proceedings of a Symposium on the Health Effects of Smokeless Tobacco

Introduction: Proceedings of a Symposium on the Health Effects of Smokeless Tobacco