Tobacco Constituents Research Articles

Synthesis of Nano-Crystalline Materials in Open-Air Laboratory: A Case Study of Tobacco.

The work deals with synthesis of nano-crystalline materials in open-air laboratory and in-depth investigation of the tobacco sample of one branded cigarette and its ash using high-resolution transmission electron microscopy and associated techniques. It exhibits the presence of nanocrystals and nanorods of various oxides in cigarette ash. The structure, shape, size and composition of these nanocrystals and nanorods are explored. The energy dispersive X-ray spectra from different regions of the tobacco sample and its ash using high-angle annular dark field scanning/transmission electron microscopy mode are utilized to obtain elemental composition and their relative abundances. For a detailed distribution of different elements in the nanorods, elemental mapping using energy-filtered transmission electron microscopy is also presented. The results highlight the conversion of amorphous constituents of tobacco to nanomaterials on combustion at low temperatures, thus mixing up in the atmosphere.

What role does tobacco smoking play in multiple sclerosis disability and mortality? A review of the evidence.

There is increasing evidence of tobacco smoking as an associative factor in multiple sclerosis (MS). Numerous studies have been conducted investigating the effects of smoking before the onset of MS as well as its impact on disease course. This special report reviews the available evidence and summarizes the contribution of smoking to increased mortality in patients with MS. It also explores some putative mechanisms for the involvement of tobacco constituents in the pathology of MS and the effects of smoking on disease-modifying treatments.

Construction of a fractional condensation device and its application in the analysis of volatile compounds from tobacco

A novel fractional condensation device was constructed for the analysis of complex volatile constituents in cut tobacco.

Evidence supporting product standards for carcinogens in smokeless tobacco products.

Smokeless tobacco products sold in the United States vary significantly in yields of nicotine and tobacco-specific nitrosamines (TSNA). With the passage of the Family Smoking Prevention and Tobacco Control Act, the Food and Drug Administration now has the authority to establish product standards. However, limited data exist determining the relative roles of pattern of smokeless tobacco use versus constituent levels in the smokeless tobacco product in exposure of users to carcinogens. In this study, smokeless tobacco users of brands varying in nicotine and TSNA content were recruited from three different regions in the U.S. Participants underwent two assessment sessions. During these sessions, demographic and smokeless tobacco use history information along with urine samples to assess biomarkers of exposure and effect were collected. During the time between data collection, smokeless tobacco users recorded the amount and duration of smokeless tobacco use on a daily basis using their diary cards. Results showed that independent of pattern of smokeless tobacco use and nicotine yields, levels of TSNA in smokeless tobacco products played a significant role in carcinogen exposure levels. Product standards for reducing levels of TSNA in smokeless tobacco products are necessary to decrease exposure to these toxicants and potentially to reduce risk for cancer.

Animal models to assess the abuse liability of tobacco products: Effects of smokeless tobacco extracts on intracranial self-stimulation

BackgroundPreclinical models are needed to inform regulation of tobacco products by the Food and Drug Administration (FDA). Typically, animal models of tobacco addiction involve exposure to nicotine alone or nicotine combined with isolated tobacco constituents (e.g. minor alkaloids). The goal of this study was to develop a model using extracts derived from tobacco products that contain a range of tobacco constituents to more closely model product exposure in humans. MethodsThis study compared the addiction-related effects of nicotine alone and nicotine dose-equivalent concentrations of aqueous smokeless tobacco extracts on intracranial self-stimulation (ICSS) in rats. Extracts were prepared from Kodiak Wintergreen, a conventional product, or Camel Snus, a potential “modified risk tobacco product”. Binding affinities of nicotine alone and extracts at various nicotinic acetylcholine receptor (nAChR) subtypes were also compared. ResultsKodiak and Camel Snus extracts contained levels of minor alkaloids within the range of those shown to enhance nicotine's behavioral effects when studied in isolation. Nonetheless, acute injection of both extracts produced reinforcement-enhancing (ICSS threshold-decreasing) effects similar to those of nicotine alone at low to moderate nicotine doses, as well as similar reinforcement-attenuating/aversive (ICSS threshold-increasing) effects at high nicotine doses. Extracts and nicotine alone also had similar binding affinity at all nAChRs studied. ConclusionsRelative nicotine content is the primary pharmacological determinant of the abuse liability of Kodiak and Camel Snus as measured using ICSS. These models may be useful to compare the relative abuse liability of other tobacco products and to model FDA-mandated changes in product performance standards.

Smokers in addiction treatment see their risk of tobacco-related disease as lower than that of an average smoker

This chapter describes the major observations and principal controversies related to the effects of cigarette smoking and the constituents of tobacco on the female and male reproduction and the development of offspring. Maternal exposure is assessed relative to specific tobacco-related chemicals and the impact of mutagenic products on reproductive targets. Maternal smoking during pregnancy is associated with adverse pregnancy outcomes, including low birth weight, premature delivery, spontaneous abortion, placental abruption, and perinatal mortality. Important new information is being learned from clinical in vitro fertilization and assisted reproduction technologies that have allowed for a more thorough analysis of the negative consequences of smoking on pregnancy cycles. In men, the effects of smoking have been reported as reductions in sperm density, motility, and function. Additional evidence links cigarette smoke exposure in utero with the development of disease pathologies in older offspring, including childhood cancers, respiratory disorders, diabetes, obesity, and developmental and behavioral consequences. Lastly, increased emphasis is placed on smoking cessation during pregnancy using pharmacological intervention with nicotine replacement.

Effects of adolescent treatment with nicotine, harmane, or norharmane in male Sprague–Dawley rats

The initiation of tobacco use occurs most often in adolescence and may be especially detrimental as the adolescent brain is undergoing substantial development. In addition to nicotine, there are over 9000 other compounds present in tobacco products, including the β-carbolines harmane and norharmane. The present study aimed to determine the long-term effects of adolescent exposure to nicotine (NIC), harmane (HAR), or norharmane (NOR) on locomotor activity, learning and memory, anxiety-like behavior, motor coordination, and monoamine/metabolite concentrations in the striatum and nucleus accumbens of male Sprague–Dawley rats. Beginning on postnatal day (PND) 27 and continuing through PND 55, subjects received twice daily intraperitoneal injections of 1ml/kg saline (CON), 0.5mg NIC/kg, 0.5mg HAR/kg, or 0.5mg NOR/kg. Body weight, food, and water intake were measured daily (PNDs 27–96). Locomotor activity was assessed on PND 40 or 41, PND 55, and PNDs 81 and 82. Other behaviors (anxiety-like behavior, motor coordination, and spatial learning and memory) were assessed at least 25days after drug exposure ended (PNDs 80–91). On PND 97, subjects were decapitated and the striatum and nucleus accumbens were dissected and frozen for analysis. NIC treatment significantly decreased food intake, but did not alter locomotor activity during or after treatment. HAR and NOR treatment, however, caused significant open field hypoactivity. Motor coordination, water maze performance, and concentrations of monoamines and metabolites in the striatum and nucleus accumbens were unaltered by any drug treatment. These results indicate a long-lasting effect on activity levels from adolescent HAR or NOR treatment; however, there were few long-lasting NIC effects. Given the paucity of data describing effects of HAR or NOR exposure, these data should encourage additional studies of these tobacco constituents as well as constituent combination studies.

Determination of behaviorally effective tobacco constituent doses in rats.

While nicotine has been established as the primary addictive drug that promotes tobacco use, recent peer-reviewed studies suggest that tobacco smoke contains additional chemical constituents that may have addictive potential. Additional research is necessary to determine the addictive potential of these tobacco constituents individually and in combination with tobacco smoke condensate; however, the behaviorally effective constituent doses necessary to conduct such studies are unclear. The primary objective of this study was to conduct behavioral studies in adult rats to determine the relevant behaviorally effective doses of the tobacco constituents, cotinine, myosmine, and anatabine to be used in future studies assessing the addictive potential of these compounds. Separate groups of adult male Sprague Dawley rats were treated with vehicle, nicotine, or various doses of cotinine, mysomine, or anatabine. Effects on locomotor activity were measured in 10-min bins for 60min. Nicotine (0.8mg/kg) produced a biphasic effect on locomotor activity, with hypoactivity during the first 10min and hyperactivity at 40-50min. In contrast, cotinine (0.1mg/kg) and myosmine (10-50mg/kg) decreased activity without a later increase. Anatabine significantly increased locomotor activity at 1mg/kg, but decreased it at 10mg/kg. Prominent effects on overt behavior were observed at anatabine doses of 10mg/kg and above. Nicotine, cotinine, myosmine, and anatabine produced distinct time- and dose-dependent patterns of effects on locomotor activity. Results from the study will aid in the selection of relevant doses for future studies assessing the addictive potential of these non-nicotine tobacco constituents.

Influence of usage time on exposure of snus users to nicotine, NNN and NNK from snus pouches

Poster POS1-23 presented at 2012 SRNT 18th Annual Meeting, March 13-16, Houston, Texas, USA. This study was entirely funded by British American Tobacco INTRODUCTION Measuring the amount of constituents extracted by snus consumers during use is a valuable step in estimating exposure to tobacco constituents. A factor potentially influencing the extent of exposure is the length of time that individual consumers keep snus pouches in their mouths during use. A 2007/2008 survey of snus use in Sweden established that, on average, use of pouched snus extends from approximately 30 minutes to just less than 120 minutes1. The objective of this study therefore was to quantify the importance of use duration on constituent transfer from pouched snus to the consumer over this timescale. The constituents selected for evaluation were nicotine and the two tobacco-specific nitrosamines (TSNAs) classified by IARC as Group 1 (sufficient evidence of carcinogenicity in humans)2, 4-(N-Methylnitrosamino)-1-(3-pyridinyl)-1-butanone (NNK) and N-Nitrosonornicotine (NNN).

Effects of tobacco smoke constituents, anabasine and anatabine, on memory and attention in female rats.

Nicotine has been well characterized to improve memory and attention. Nicotine is the primary, but not only neuroactive compound in tobacco. Other tobacco constituents such as anabasine and anatabine also have agonist actions on nicotinic receptors. The current study investigated the effects of anabasine and anatabine on memory and attention. Adult female Sprague-Dawley rats were trained on a win-shift spatial working and reference memory task in the 16-arm radial maze or a visual signal detection operant task to test attention. Acute dose-effect functions of anabasine and anatabine over two orders of magnitude were evaluated for both tasks. In the radial-arm maze memory test, anabasine but not anatabine significantly reduced the memory impairment caused by the NMDA antagonist dizocilpine (MK-801). In the signal detection attentional task, anatabine but not anabasine significantly attenuated the attentional impairment caused by dizocilpine. These studies show that non-nicotine nicotinic agonists in tobacco, similar to nicotine, can significantly improve memory and attentional function. Both anabasine and anatabine produced cognitive improvement, but their effectiveness differed with regard to memory and attention. Follow-up studies with anabasine and anatabine are called for to determine their efficacy as therapeutics for memory and attentional dysfunction.

Reinforcing and neural activating effects of norharmane, a non-nicotine tobacco constituent, alone and in combination with nicotine

Tobacco use is the leading cause of preventable death. Although the health risks are well known, cessation rates remain low. Whereas behavioral and neuroanatomical studies on tobacco addiction conventionally use nicotine, there is evidence that other constituents, such as monoamine oxidase inhibitors, may be important factors for modeling smoking. The aims of the present study were therefore to determine whether norharmane, a tobacco constituent and monoamine oxidase inhibitor, is self-administered alone and/or in combination with nicotine, and to evaluate the neural mechanisms underlying acquisition of self-administration of the two drugs. Sprague–Dawley rats were catheterized and allowed to intravenously self-administer either saline, nicotine (7.5 μg/kg/inj), norharmane (0.25 or 2.5 μg/kg/inj), alone or combined together (7.5 + 2.5 μg/kg/inj) for five days at fixed ratio (FR)1, two days each at FR2 and FR5, and one day at progressive ratio. Animals acquired self-administration of norharmane alone (2.5 μg/kg/inj), and the reinforcing effects of nicotine and norharmane were additive. For neuroanatomical analyses, rats self-administered the same treatments for six days at FR1, then brains were collected and processed by in situ hybridization for cfos mRNA expression. Treatment-specific profiles of regional cfos expression and correlations between cfos mRNA levels and behavioral responding were observed. Thus, not only was norharmane behaviorally reinforcing but, when combined with nicotine, resulted in patterns of neural activation distinct from that of norharmane or nicotine alone. This suggests that non-nicotine constituents can have central activating effects independent of nicotine, further substantiating the need for their inclusion in preclinical investigations of tobacco dependence.

Occupational Health Problems amongst Women Beedi Rollers in Jhansi, Bundelkhand Region, Uttar Pradesh

The aim of this study was to investigate the prevalence of occupationally related health problems among women beedi rollers in Jhansi, Uttar Pradesh, Bundelkhand region, India. Beedi rolling is a serious occupational health hazard as these workers are constantly exposed to tobacco dust, fumes and other dangerous chemicals viz: nitrosamines and nicotine which are readily absorbed by the body through skin, respiratory epithelium and mucous membrane of mouth, nose and intestine. Exposure to tobacco dust is known to affect the respiratory tracts in humans. Present study revealed that women beedi rollers face numerous health problems possibly due to direct inhalation of tobacco flakes. This study included 216 females (mean age 39.17 ± 11.95 Years) actively involved in beedi rolling to ascertain the effect of beedi rolling on health. The study found that majority of the respondents complained problems i.e. joint pain, eye problems, Nervous and skin diseases. Increased systemic exposure to tobacco constituents was evident from the high levels of creatinine in urine samples. Thus beedi rolling causes serious health problems in women.

Pictorial health warning label content and smokers' understanding of smoking-related risks-a cross-country comparison.

The aim of the present study was to assess smokers' level of agreement with smoking-related risks and toxic tobacco constituents relative to inclusion of these topics on health warning labels (HWLs). 1000 adult smokers were interviewed between 2012 and 2013 from online consumer panels of adult smokers from each of the three countries: Australia (AU), Canada (CA) and Mexico (MX). Generalized estimating equation models were estimated to compare agreement with smoking-related risks and toxic tobacco constituents. For disease outcomes described on HWLs across all three countries, there were few statistical differences in agreement with health outcomes (e.g. emphysema and heart attack). By contrast, increases in agreement where the HWLs were revised or introduced on HWLs for the first time (e.g. blindness in AU and CA, bladder cancer in CA). Similarly, samples from countries that have specific health content or toxic constituents on HWLs showed higher agreement for that particular disease or toxin than countries without (e.g. higher agreement for gangrene and blindness in AU, higher agreement for bladder cancer and all toxic constituents except nitrosamines and radioactive polonium in CA). Pictorial HWL content is associated with greater awareness of smoking-related risks and toxic tobacco constituents.

The role of zinc, copper, plasma glutathione peroxidase enzyme, and vitamins in the development of allergic diseases in early childhood: The Polish mother and child cohort study.

It has been hypothesized that the increase in allergic disorders may, in part, be a consequence of changing diet. The primary aim of this study was to assess the associations between occurrence of atopic dermatitis; food allergy; the incidence of wheeze inhaled glucocorticosteroid use in children during the 1st year of life; and cord blood concentrations of copper, zinc, vitamins (A and E), and glutathione peroxidase activity. We evaluated 240 1-year-old children from the Polish Mother and Child Cohort Study. Women were interviewed during pregnancy to collect demographic and socioeconomic data and medical and reproductive history. Exposure to tobacco constituents was assessed based on questionnaire data. At delivery, umbilical cord blood plasma was sampled. One year after the birth, the child's exposure and health status were examined. In the analyses a multivariable model was used. Higher zinc and copper concentrations in cord blood were associated with increased likelihood of wheezing in 1-year-old children. This effect was seen only among children exposed to tobacco smoke at home. We also showed significantly lower activity of glutathione peroxidase enzyme 3 in umbilical cord blood plasma of children with atopic dermatitis during the 1st year of life. There were no significant associations between vitamin A and E concentrations in plasma and children's health. We showed imbalance in the antioxidant defense system in cord blood, which may lead to development of atopic dermatitis or wheezing in infancy. The association between maternal nutrient status during pregnancy and child's health is complex and interacts with other environmental factors such as tobacco exposure. This study was a part of the clinical trial NCT01861548 registered at ClinicalTrials.gov.

The effect of nicotine on osteoinduction by recombinant human bone morphogenetic protein 2

Nicotine, one of the constituents of tobacco, is known to have an adverse effect on human health. We sought to clarify the interaction between nicotine and recombinant human bone morphogenetic protein 2 (rhBMP-2) in terms of osteogenesis in vitro and osteoinduction in vivo. Nicotine did not inhibit or stimulate alkaline phosphatase (ALP) activity or the amount of osteocalcin in C2C12 cells in the presence of rhBMP-2 in vitro. Ectopic bone formation using a collagen sponge containing rhBMP-2 was evaluated with and without nicotine after 21 days using radiographic, histological, biochemical, and immunohistochemical analyses. ALP activity in the medium-dose group (2.2±0.9IU/mg protein; P=0.047) and the high-dose group (2.0±0.1IU/mg protein; P=0.03) was significantly lower than in the control group. The calcium content in the medium-dose group (35.4±12.9μg/mg tissue; P=0.0099) and high-dose group (34.8±10.5μg/mg tissue; P=0.006) was significantly lower than in the control group. The number of vascular endothelial growth factor-positive cells in the high-dose group (671.9±57.3cells/mm2; P=0.03) was significantly lower than in the control group. Results showed that nicotine did not inhibit the stimulatory effect of rhBMP-2 in vitro, but a high dose of nicotine inhibited bone formation in vivo by adversely affecting vascularization.

Monitoring Tobacco-Specific N-Nitrosamines and Nicotine in Novel Smokeless Tobacco Products: Findings From Round II of the New Product Watch

Analysis of novel smokeless tobacco products purchased in Round I of the New Product Watch (NPW)-a national tobacco monitoring network-demonstrated that some tobacco constituents vary not only across various brands but also regionally and over time within the same product. In this study, we analyzed snus and dissolvable tobacco products that were purchased in Round II of the NPW. We analyzed tobacco-specific N-nitrosamines (TSNA) and nicotine in snus and dissolvable tobacco products that were purchased in various regions of the country during the spring and summer of 2011. The results were compared against the Round I data, across different U.S. regions, and among products. A total of 216 samples were received from different states representing 6 regions of the country. Compared with the previous analyses, TSNA levels increased significantly in Marlboro and Camel Snus and some dissolvable Camel products. The levels of unprotonated nicotine in Marlboro Snus and Camel Snus in this study were not different from Round I but varied significantly by regions; the differences between the highest and the lowest average regional levels were ~3.2-fold in Marlboro Snus ~1.7-fold in Camel Snus. Our results indicate that some novel smokeless tobacco products contain TSNA at the levels found in the conventional moist snuff. Observation of regional variations in unprotonated nicotine content in both Round I and Round II of NPW suggest that manufacturers may tailor the levels of this constituent consistently to different regions.

Differential effects of non-nicotine tobacco constituent compounds on nicotine self-administration in rats

Tobacco smoking has been shown to be quite addictive in people. However, nicotine itself is a weak reinforcer compared to other commonly abused drugs, leading speculation that other factors contribute to the high prevalence of tobacco addiction in the human population. In addition to nicotine, there are over 5000 chemical compounds that have been identified in tobacco smoke, and more work is needed to ascertain their potential contributions to tobacco's highly addictive properties, or as potential candidates for smoking cessation treatment. In this study, we examined seven non-nicotine tobacco constituent compounds (anabasine, anatabine, nornicotine, myosmine, harmane, norharmane, and tyramine) for their effects on nicotine self-administration behavior in rats. Young adult female Sprague–Dawley rats were allowed to self-administer nicotine (0.03mg/kg/50μl infusion) under a fixed ratio-1 schedule of reinforcement. Each self-administration session lasted 45min. Doses of each tobacco constituent compound were administered subcutaneously 10min prior to the start of each session in a repeated measures, counterbalanced order two times. Anabasine displayed a biphasic dose–effect function. Pretreatment with 0.02mg/kg anabasine resulted in a 25% increase in nicotine self-administration, while 2.0mg/kg of anabasine reduced nicotine infusions per session by over 50%. Pretreatment with 2.0mg/kg anatabine also significantly reduced nicotine self-administration by nearly half. These results suggest that some non-nicotine tobacco constituents may enhance or reduce nicotine's reinforcing properties. Also, depending upon the appropriate dose, some of these compounds may also serve as potential smoking cessation agents.

Nicotine Exposure, Blood Pressure, and Inflammation in Tobacco Smokers and Chewers in a Rural Community in Nepal

Tobacco consumption is high amongst the people of Nepal. This study was carried out in 2011 in a rural community of Nepal, to compare pathological parameters associated with tobacco use in relation to nicotine metabolism between smokers, chewers, and a control group. A total of 216 volunteers provided blood and urine samples for testing nicotine metabolites, C-reactive protein, and cell counts. Data were analyzed using ANOVA, correlation, and t-tests using STATA. Differences in blood pressure amongst the groups indicate a role of smoking in preventing a rise in BP with age, likely attributable to a different mechanism of metabolism of tobacco constituents.

Simple Determination of Sugars in Cigarettes

Simple sugars are an important constituent of tobacco, varying between almost 0 and 20% on weight of tobacco. Some manufacturers add sugars to the natural content in tobacco. In the present report, a simple and fast method is presented to analyze the concentrations of the sugars sucrose, glucose and fructose in cigarettes. After an extraction step, the diluted samples are applied to an isocratic HPLC system with detection by evaporated light scattering (ELSD). The method is very stable and sensitive with a good reproducibility. In one working day 80 samples of cigarettes can be processed and prepared for overnight HPLC analysis by one technician. The average total sugar content in 58 commercial cigarette brands is 17.4% (w/w) with a range of 1.9 to 18.3%.

Effect of Nicotine-free Tobacco Extract on DNA Damage Responses in Cancerous and Non-cancerous Cells

The use of cigarette products tags tobacco as a crop with serious health concerns, but nutraceutical potentials of tobacco constituents other than nicotine have not been adequately explored. The objective of this study was to investigate the effect of nicotine-free tobacco extract on the activation of tumorigenesis barriers. Four lines of human cells were treated with a nicotine-free tobacco extract (0.1 and 1 mg/mL, 0-48 h). MRC-5 and CCD841 non-cancerous cells treated with the tobacco extract showed hypersensitivity, apoptosis and cell cycle arrest in S and G2/M phases, and induction of DNA damage response as evidenced by phosphorylation of ataxia telangiectasia mutated at Ser- 1981 and histone H2A.X at Ser-139. In contrast, HCT 116 cancerous cells, with or without functional DNA mismatch repair, were resistant to the tobacco extract treatment. These results suggest that tobacco components other than nicotine may have a chemoprevention potential that stifles tumorigenesis at the early stage of tumorigenesis