Tobacco experimental model to induce urinary bladder neoplasms

OBJECTIVE: to develop an experimental model of exposure to tobacco burning (cigarette) products to assess the effects of its chronic use in relation to cancers of the bladder. METHODS: the animals were chronically exposed to the burning tobacco products in a semi-open chamber to simulate smoking. Thirty young Wistar rats were divided into two groups: one with 20 animals simulating smoking for six months, and ten not exposed control animals for the same period. After exposure by inhalation of cigarette smoke, animals were euthanized and subjected to histopathological study of the bladder wall. RESULTS: no tumor was found but mild and non significant alterations. The studies of hemo-oximetry (carboxyhemoglobin and methemoglobin) and the concentration of carbon dioxide (CO2) confirm that the animals were exposed to high concentrations of tobacco smoke and its derivatives. CONCLUSION: no bladder mucosal neoplasia was found in the pathological study of animals. The developed experimental models were highly efficient, practical and easy to use and can be used in other similar studies to determine the harmful effects caused by smoking.

A tobacco cDNA reveals two different transcription patterns in vegetative and reproductive organs

In order to identify genes expressed in the pistil that may have a role in the reproduction process, we have established an expressed sequence tags project to randomly sequence clones from a Nicotiana tabacum stigma/style cDNA library. A cDNA clone (MTL-8) showing high sequence similarity to genes encoding glycine-rich RNA-binding proteins was chosen for further characterization. Based on the extensive identity of MTL-8 to the RGP-1a sequence of N. sylvestris, a primer was defined to extend the 5' sequence of MTL-8 by RT-PCR from stigma/style RNAs. The amplification product was sequenced and it was confirmed that MTL-8 corresponds to an mRNA encoding a glycine-rich RNA-binding protein. Two transcripts of different sizes and expression patterns were identified when the MTL-8 cDNA insert was used as a probe in RNA blots. The largest is 1,100 nucleotides (nt) long and markedly predominant in ovaries. The smaller transcript, with 600 nt, is ubiquitous to the vegetative and reproductive organs analyzed (roots, stems, leaves, sepals, petals, stamens, stigmas/styles and ovaries). Plants submitted to stress (wounding, virus infection and ethylene treatment) presented an increased level of the 600-nt transcript in leaves, especially after tobacco necrosis virus infection. In contrast, the level of the 1,100-nt transcript seems to be unaffected by the stress conditions tested. Results of Southern blot experiments have suggested that MTL-8 is present in one or two copies in the tobacco genome. Our results suggest that the shorter transcript is related to stress while the larger one is a flower predominant and nonstress-inducible messenger.

Induction of a gloverin-like antimicrobial polypeptide in the sugarcane borer Diatraea saccharalis challenged by septic injury

Diatraea saccharalis (Fabricius, 1794) (Lepidoptera: Crambidae) is an important pest for Brazilian sugarcane. In the present study, we detected two distinct spots in hemolymph from septic injured larvae (HDs1 and HDs2), which are separated by 2DE gel electrophoresis. Both spots were subjected to in-gel tryptic digestion and MALDI-TOF/TOF analysis, which revealed the sequence VFGTLGSDDSGLFGK present in both HDs1 and HDs2. This sequence had homology and 80% identity with specific Lepidoptera antimicrobial peptides called gloverins. Analyses using the ImageMaster 2D software showed pI 8.94 of the HDs1 spot, which is similar to that described to Hyalophora gloveri gloverin (pI 8.5). Moreover, the 14-kDa molecular mass of the spot HDs1 is compatible to that of gloverins isolated from the hemolymph of Trichoplusia ni, Helicoverpa armigera and H. gloveri. Antimicrobial assays with partially purified fractions containing the HDs1 and HDs2 polypeptides demonstrated activity against Escherichia coli. This is the first report of antimicrobial polypeptides in D. saccharalis, and the identification of these peptides may help in the generation of new strategies to control this pest.

Education, tobacco smoking, alcohol consumption, and IL-2 and IL-6 gene polymorphisms in the survival of head and neck cancer

The association of education, tobacco smoking, alcohol consumption, and interleukin-2 (IL-2 +114 and -384) and -6 (IL-6 -174) DNA polymorphisms with head and neck squamous cell carcinoma (HNSCC) was investigated in a cohort study of 445 subjects. IL-2 and IL-6 genotypes were determined by real-time PCR. Cox regression was used to estimate hazard ratios (HR) and 95% confidence intervals (95%CI) of disease-specific survival according to anatomical sites of the head and neck. Mean age was 56 years and most patients were males (87.6%). Subjects with 5 or more years of schooling had better survival in larynx cancer. Smoking had no effect on HNSCC survival, but alcohol consumption had a statistically significant effect on larynx cancer. IL-2 gene +114 G/T (HR = 0.52; 95%CI = 0.15-1.81) and T/T (HR = 0.22; 95%CI = 0.02-3.19) genotypes were associated with better survival in hypopharynx cancer. IL-2 +114 G/T was a predictor of poor survival in oral cavity/oropharynx cancer and larynx cancer (HR = 1.32; 95%CI = 0.61-2.85). IL-2 -384 G/T was associated with better survival in oral cavity/oropharynx cancer (HR = 0.80; 95%CI = 0.45-1.42) and hypopharynx cancer (HR = 0.68; 95%CI = 0.21-2.20), but an inverse relationship was observed for larynx cancer. IL-6 -174 G/C was associated with better survival in hypopharynx cancer (HR = 0.68; 95%CI = 0.26-1.78) and larynx cancer (HR = 0.93; 95%CI = 0.42-2.07), and C/C reduced mortality in larynx cancer. In general, our results are similar to previous reports on the value of education, smoking, alcohol consumption, and IL-2 and IL-6 genetic polymorphisms for the prognosis of HNSCC, but the risks due to these variables are small and estimates imprecise.

Expression of NR1I3 in mouse lung tumors induced by the tobacco-specific nitrosamine 4-(methylnitrosamino)-4-(3-pyridyl)-1-butanone

Nuclear receptor subfamily 1, group I, member 3 (NR1I3) is reported to be a possible novel therapeutic target for some cancers, including lung, brain and hematopoietic tumors. Here, we characterized expression of NR1I3 in a mouse model of lung carcinogenesis induced by 4-(methylnitrosamino)-4-(3-pyridyl)-1-butanone (NNK), the most potent tobacco carcinogen. Lung tumors were collected from mice treated with NNK (400 mg/kg) and euthanized after 52 weeks. Benign and malignant lesions were formalin-fixed and paraffin-embedded for histology and immunohistochemistry, with samples snap-frozen for mRNA analysis. Immunohistochemically, we found that most macrophages and type I and II pneumocytes expressed NR1I3, whereas fibroblasts and endothelial cells were NR1I3−. Compared with benign lesions, malignant lesions had less NR1I3+ tumor cells. Gene expression analysis also showed an inverse correlation between NR1I3 mRNA expression and tumor size (P=0.0061), suggesting that bigger tumors expressed less NR1I3 transcripts, in accordance with our immunohistochemical NR1I3 tests. Our results indicate that NR1I3 expression decreased during progression of malignant lung tumors induced by NNK in mice.

A qualitative case study of the processes of peer education in a young adult tobacco control initiative, Leave the Pack Behind

Peer education involves peers offering credible and reliable information about sensitive life issues through the means of an informal peer group setting (Topping & Ehly, 1998). The purpose of this instrumental case study was to examine the processes of peer education through the exploration of two teams within a young adult tobacco control initiative, Leave the Pack Behind (LTPB). This qualitative case study examined two peer education teams over an eight-month period. Interviews, focus groups and observations were conducted with 12 participants across two peer education teams. Findings show the complexities of the processes of peer education including a connection between the stages of change and the changing role of the peer educator across stages of the empowerment process. Peer education teams and factors in the macro environment were also found to impact the process of peer education. This study provides a new definition for the process of peer education: peer education is a fluid process of knowledge exchange in which peer educators adopt different styles of facilitation as people move through stages of empowerment and change. This study contributes to the academic hterature upon the processes of peer education by providing a definition, a model and an overall understanding through an ecological and empowerment framework. The findings from this study suggest peer educators can be further trained to: use specific peer educational approaches that fit with student smoker's stage of change; better understand their position as a peer educator on the LTPB team; understand the reciprocal relationship between the macro environment and the peer education teams having an effect on one another.

Students' knowledge about tobacco one year after receiving the PAL smoking prevention program

A review of literature revealed that the control of cigarette smoking could do more to improve health than any other single action in the field of preventive medicine. In Ontario, since 1989, both Public Health Units and Boards ofEducations have been mandated to provide educational studies related to tobacco use prevention. Even given this fact, there has been an increase in smoking behaviQurs at an earlier age and in females in particular. Smoking prevention progralns must use the most effective means to assist students to obtain the knowledge and skills required to remain or becom'e nonsmokers. In the Niagara Region, PAL smoking prevention programs are offered in some, but not all, schools. As a form of program evaluation, this research sought to determine if students who had PAL could answer correctly a greater number of smoking-related questions than students who did not have this program. Findings reported that students who had PAL in Grade 6 were able to correctly answer more knowledge-based questions (at a statistically significant level), could provide ways to refuse cigarettes at a greater rate, and were able to provide more reasons for remaining nonsmokers. Students who had smoking prevention programming re,ported smoking behavio'urs at a lower rate than those who did not receive this type of program.

Biochemical and histological investigations of viral localisation in the hypersensitive reaction of Phaselous vulgaris L. var Pinto to tobacco mosaic virus infection

STOBBS, Lorne,W ABSTRACT Biochemical and Histological Investigations of viral localisation in the hypersensitive reaction of Phaseolus vulgaris L. var Pinto to tobacco mosaic virus infection. The infection of Phaseolus vulgaris L. var Pinto with tobacco mosaic virus (TMV) results in the production of distinct necrotic lesions confining the virus to restricted areas of the leaf surface. Biochemical and histological changes in the leaf tissue as a result of infection have been described. Trace accumulations of fluorescent metabolites, detected prior to lesion expression represent metabolites produced, by the cell in response to virus infection. These substances, are considered to undergo oxidation and in diffusing into adjacent cells, react with cellular constituents causing the death of these cells. Such cellular necrosis in advance of infection effectively limits virus spread. Chromatographic studies on extracts from TMV infected Pinto bean leaf tissue suggests that a number of extra-fluorescent metabolites produced on lesion'expression represent end products of phenolic oxidation r,eactionsoccurring earlier in these cells. Inhibition of phenolic oxidation by ascorbate infiltration or elevated temperature treatment resulted in the absence of extra-fluorescent metabolites and the continued movement of virus in the absence of necrosis. Further studies with i ascorbate infiltration indicated that irreversible necrotic events were determined as early as 12 tci 18 hrs after viral inoculation. Histochemical tests indicated that callose formation was initiated at this time, and occurred in response to necrotisation. Inhibition of necrosis by either ascorbate infiltration or elevated temp8rature treatment resulted in the absence of callose deposition. Scanning electron'micrographs of infected tissue revealed severe epidermal and palisade cell damage. Histochemical tests indicated extensive callose formation in cells bordering the lesion, and suggested the role of callose iTh the blockage of intercellular connections limiting virus movement. The significance of these cellular changes is discussed. ii

The impact of tobacco control policies on university students' smoking in Ontario

Objective. Despite steady declines in the prevalence of tobacco use among Canadians, young adult tobacco use has remained stubbornly high over the past two decades (CTUMS, 2005a). Currently in Ontario, young adults have the highest proportion of smokers of all age cohorts at 26%. A growing body of evidence shows that smoking restrictions and other tobacco control policies can reduce tobacco use and consumption among adults and deter initiation among youth; whether young adult university students' smoking participation is influenced by community smoking restrictions, campus tobacco control policies or both remains an empirical question. The purpose of this study is to examine the relationship among current smoking status of students on university campuses across Ontario and various tobacco control policies, 3including clean air bylaws of students' home towns, clean air by-laws of the community where the university is situated, and campus policies. Methods. Two data sets were used. The 200512006 Tobacco Use in a Representative Sample of Post-Secondary Students data set provides information about the tobacco use of 10,600 students from 23 universities and colleges across Ontario. Data screening for this study reduced the sample to 5,114 17-to-24 year old undergraduate students from nine universities. The second data set is researcher-generated and includes information about strength and duration of, and students' exposure to home town, local and campus tobacco control policies. Municipal by-laws (of students' home towns and university towns) were categorized as weak, moderate or strong based on criteria set out in the Ontario Municipal By-law Report; campus policies were categorized in a roughly parallel fashion. Durations of municipal and campus policies were calculated; and length of students' exposure to the policies was estimated (all in months). Multinomial logistic regression analyses were used to examine the relationship between students' current smoking status (daily, less-than-daily, never-smokers) and the following policy measures: strength of, duration of, and students' exposure to campus policy; strength of, duration of, and students' exposure to the by-law in the university town; and, strength of, duration of, and students' exposure to the by-law in the home town they grew up in. Sociodemographic variables were controlled for. Results. Among the Ontario university students surveyed, 7.0% currently use tobacco daily and 15.4% use tobacco less-than-daily. The proportions of students experiencing strong tobacco control policies in their home town, the community in which their university is located and at their current university were 33.9%,64.1 %, and 31.3% respectively. However, 13.7% of students attended a university that had a weak campus policy. Multinomial logistic regressions suggested current smoking status was associated with university town by-law strength, home town by-law strength and the strength of the campus tobacco control policy. In the fmal model, after controlling for sociodemographic factors, a strong by-law in the university town and a strong by-law in students' home town were associated with reduced odds of being both a less-than-daily (OR = 0.64, 95%CI: 0.48-0.86; OR = 0.80, 95%CI: 0.66-0.95) and daily smoker (OR = 0.59, 95%CI: 0.39-0.89; OR = 0.76, 95%CI: 0.58-0.99), while a weak campus tobacco control policy was associated with higher odds of being a daily smoker (OR = 2.08, 95%CI: 1.31-3.30) (but unrelated to less-than-daily smoking). Longer exposure to the municipal by-law (OR = 0.93; 95%CI: 0.90-0.96) was also related to smoking status. Conclusions. Students' smoking prevalence was associated with the strength of the restrictions in university, and with campus-specific tobacco control policies. Lessthan- daily smoking was not as strongly associated with policy measures as daily smoking was. University campuses may wish to adopt more progressive campus policies and support clean air restrictions in the broader community. More research is needed to determine the direction of influence between tobacco control policies and students' smoking.

Contraband tobacco on post-secondary campuses in Ontario

In Ontario 27% of young adults smoke, and annual surveillance data suggests tobacco use is plateauing after years of decline. The availability of inexpensive contraband tobacco products maybe contributing to this situation. Limited research has been conducted on the use of contraband tobacco and despite the increasing availability of contraband 'Native cigarettes', no studies to date have examined their use among young adults. Accordingly, this study examines: (a) what proportion of cigarette butts discarded on post-secondary campuses are contraband; and (b) whether the proportion of contraband butts varies between colleges and universities, across seven geographical regions in the province and based on proximity First Nations reserves. In March and April 2009, discarded cigarette butts were collected from the grounds of 25 post-secondary institutions across Ontario. At each school, cigarette butts were collected on a single day from four locations. The collected cigarette butts were reliably sorted into five categories according to their filtertip logos: legal, contraband First NationslNative cigarettes, international and suspected counterfeit cigarettes, unidentifiable and unknown. Contraband use was apparent on all campuses, but varied considerably from school to school. Data suggest that contraband Native cigarettes account for as little as 1 % to as much as 38 % of the total cigarette consumption at a particular school. The highest proportion of contraband was found on campuses in the Northern part of the province. Consumption of Native contraband was generally higher on colleges compared to universities. The presence of contraband tobacco on all campuses suggests that strategies to reduce smoking among young adults must respond to this cohort's use of these products.

Patterns and correlates of tobacco use among young adults at college and university

This thesis describes college and university students' smoking behaviours and examines whether socioenvironmental and personal characteristics experienced during adolescence are differentially associated with their smoking participation. Results show more college students than university students currently smoke (37% and 21 % respectively) and more began smoking prior to post-secondary school (93% and 84% respectively). Early age of onset of alcohol use increased the odds of current smoking (main effect model, OR = 8.56 CI = 6.47, 11.33), especially for university students (interaction effect model, b = 2.35 CI = 7.50, 14.64). Lower levels of high school connectedness were associated with increased odds of current smoking but for university students only (interaction effect model, b = -0.15 CI = 0.84, 0.88). While limitations associated with convenience sampling and low response rate exist, this is the first Canadian study to examine college and university students separately. I t reveals that tobacco control programming needs to differ for college and university students, and early alcohol prevention and school engagement programs for adolescents may influence tobacco use. Given that both educational pathway and use of tobacco are associated with SES, future research may consider examining in more detail, SES-related socioenvironmental variables.

The Association between Tobacco Control Policies and Marijuana Use among Ontario Undergraduate Students

Background: Research indicates a steady increase in marijuana use and that it is concurrent with tobacco. There is speculation this concurrency reaches beyond use, to where policies aimed at reducing one may result in the reduction of the other. Purpose: To investigate the association between tobacco control policies and marijuana use among young adult undergraduates. Methods: A stratified sample of Ontario universities resulted in a sample of 4,966 participants. Results: Campuses with a moderately strong policy was found to be significantly associated with decreased marijuana use compared to campuses with a weak tobacco control policy. (OR=0.52, 95% CI: 0.36-0.76). Conclusions: The findings show tobacco control strategies are related to decreased odds of marijuana use among Ontario undergraduates. These findings are important to both policy makers and researchers interested in health strategies pertaining to marijuana and tobacco use and/or how health policies aimed at reducing one risk behaviour can affect another.

The Death of Brock at Queenston Heights, 1812: Imperial Tobacco Company silk card

General Isaac Brock was a British Army officer and administrator who was promoted to Major General. He was responsible for defending Upper Canada against the United States. He died at the Battle of Queenston Heights during the War of 1812. Bartholome Houde and George E. Tuckett (Tuckett was mayor of Hamilton Ontario in 1896) manufactured and sold tobacco before Confederation. In 1841, the company was called B. Houde and Grothe. When Houde retired in 1822 his son in law, Francis Xavier Dussault took over the company. In 1903, the company was called B. Houde and Company Ltee. and it was run by Dussault’s two sons J.A. Dussault and J.E. Dussault. The B. Houde Company had become part of American Tobacco Ltd. which merged with Empire Tobacco Co. Ltd. to form the Imperial Tobacco Company in 1908 in the St. Henri district of Montreal. In 2008, the company celebrated their centennial. Cigarette silks were either enclosed in packets of cigarettes or were redeemable by mail. People would sew these silks together to create quilts or table covers. Some cigarette packages even included instructions for making these items.

Minimal Contact Intervention for Tobacco Dependence

This study was conducted to measure the degree of adherence by public health care providers to a policy that requires them to implement minimal contact intervention for tobacco cessation with their clients. This study also described what components of the intervention may have contributed to the adherence of the policy and how health care providers felt about adhering to the policy. The intervention consisted of a policy for implementation of minimal contact intervention, changes to documentation, a health care provider mentor trained, a training session for health care providers, and ongoing paper and people supports for implementation. Data for this study were collected through a health care provider questionnaire, focus group interviews, and a compliance protocol including a chart audit. The findings of this study showed a high degree of adherence to the policy, that health care providers thought minimal contact intervention was important to conduct with their clients, and that health care providers felt supported to implement the intervention. No statistically significant difference was found between new and experienced health care providers on 17 of the 18 questions on the health care provider questionnaire. However there was a statistically significant difference between new and experienced health care providers with respect to their perception that “clients often feel like they have to accept tobacco cessation information from me.” Changes could be made to the minimal contact intervention and to documentation of the intervention. Implications for future research include implementation within other programs within Hamilton Public Health Services and implementation of this model within other public health units and other types of health care providers within Ontario.

Fractionation, chemical and toxicological characterization of tobacco smoke components

La fumée du tabac est un aérosol extrêmement complexe constitué de milliers de composés répartis entre la phase particulaire et la phase vapeur. Il a été démontré que les effets toxicologiques de cette fumée sont associés aux composés appartenant aux deux phases. Plusieurs composés biologiquement actifs ont été identifiés dans la fumée du tabac; cependant, il n’y a pas d’études démontrant la relation entre les réponses biologiques obtenues via les tests in vitro ou in vivo et les composés présents dans la fumée entière du tabac. Le but de la présente recherche est de développer des méthodes fiables et robustes de fractionnement de la fumée à l’aide de techniques de séparation analytique et de techniques de détection combinés à des essais in vitro toxicologiques. Une étude antérieure réalisée par nos collaborateurs a démontré que, suite à l’étude des produits de combustion de douze principaux composés du tabac, l’acide chlorogénique s’est avéré être le composé le plus cytotoxique selon les test in vitro du micronoyau. Ainsi, dans cette étude, une méthode par chromatographie préparative en phase liquide a été développée dans le but de fractionner les produits de combustion de l’acide chlorogénique. Les fractions des produits de combustion de l’acide chlorogénique ont ensuite été testées et les composés responsables de la toxicité de l’acide chlorogénique ont été identifiés. Le composé de la sous-fraction responsable en majeure partie de la cytoxicité a été identifié comme étant le catéchol, lequel fut confirmé par chromatographie en phase liquide/ spectrométrie de masse à temps de vol. Des études récentes ont démontré les effets toxicologiques de la fumée entière du tabac et l’implication spécifique de la phase vapeur. C’est pourquoi notre travail a ensuite été focalisé principalement à l’analyse de la fumée entière. La machine à fumer Borgwaldt RM20S® utilisée avec les chambres d’exposition cellulaire de British American Tobacco permettent l’étude in vitro de l’exposition de cellules à différentes concentrations de fumée entière du tabac. Les essais biologiques in vitro ont un degré élevé de variabilité, ainsi, il faut prendre en compte toutes les autres sources de variabilité pour évaluer avec précision la finalité toxicologique de ces essais; toutefois, la fiabilité de la génération de la fumée de la machine n’a jamais été évaluée jusqu’à maintenant. Nous avons donc déterminé la fiabilité de la génération et de la dilution (RSD entre 0,7 et 12 %) de la fumée en quantifiant la présence de deux gaz de référence (le CH4 par détection à ionisation de flamme et le CO par absorption infrarouge) et d’un composé de la phase particulaire, le solanesol (par chromatographie en phase liquide à haute performance). Ensuite, la relation entre la dose et la dilution des composés de la phase vapeur retrouvée dans la chambre d’exposition cellulaire a été caractérisée en utilisant une nouvelle technique d’extraction dite par HSSE (Headspace Stir Bar Sorptive Extraction) couplée à la chromatographie en phase liquide/ spectrométrie de masse. La répétabilité de la méthode a donné une valeur de RSD se situant entre 10 et 13 % pour cinq des composés de référence identifiés dans la phase vapeur de la fumée de cigarette. La réponse offrant la surface maximale d’aire sous la courbe a été obtenue en utilisant les conditions expérimentales suivantes : intervalle de temps d’exposition/ désorption de 10 0.5 min, température de désorption de 200°C pour 2 min et température de concentration cryogénique (cryofocussing) de -75°C. La précision de la dilution de la fumée est linéaire et est fonction de l’abondance des analytes ainsi que de la concentration (RSD de 6,2 à 17,2 %) avec des quantités de 6 à 450 ng pour les composés de référence. Ces résultats démontrent que la machine à fumer Borgwaldt RM20S® est un outil fiable pour générer et acheminer de façon répétitive et linéaire la fumée de cigarette aux cultures cellulaires in vitro. Notre approche consiste en l’élaboration d’une méthodologie permettant de travailler avec un composé unique du tabac, pouvant être appliqué à des échantillons plus complexes par la suite ; ex : la phase vapeur de la fumée de cigarette. La méthodologie ainsi développée peut potentiellement servir de méthode de standardisation pour l’évaluation d’instruments ou de l’identification de produits dans l’industrie de tabac.