Análise proteômica da saliva de usuários de cigarro eletrônico / Proteomic analysis of electronic cigarette users' saliva

O cigarro eletrônico, também conhecido como e-cig, tem sido amplamente utilizado nos últimos anos, principalmente pela população mais jovem. Substâncias citotóxicas e carcinogênicas podem estar presentes em sua composição, no entanto, pouco se sabe sobre os riscos do seu uso. O objetivo deste trabalho foi avaliar o perfil proteico da saliva de usuários de cigarro eletrônico por meio de estudo proteômico e as possíveis alterações salivares desses indivíduos. Para tanto, os participantes foram divididos em dois grupos: Grupo Cigarro Eletrônico (GCE), composto por 25 vaporizadores regulares e exclusivos de e-cig, e Grupo Controle (GCO), constituído por 25 indivíduos não fumantes e não vaporizadores de e-cig, pareados por sexo e idade ao GCE. Todos os participantes foram submetidos a exame clínico e coleta de saliva não estimulada para verificação da sialometria e avaliação do proteoma salivar. As amostras foram aliquotadas e submetidas às seguintes análises: viscosidade salivar, pH, capacidade tampão e mensuração da cotinina salivar realizada por meio do teste imunoenzimático (ELISA) utilizando Cotinine Enzyme Immunoassay Kit (Salimetrics). O perfil etílico dos pacientes foi avaliado por meio do teste AUDIT (Alcohol Use Disorders Identification Test) e por perguntas referentes ao consumo de álcool. Já o perfil de consumo do e-cig foi traçado por meio de perguntas referentes ao uso dos dispositivos e teste de concentração de monóxido de carbono no ar expirado. A análise proteômica foi realizada utilizando a metodologia shotgun e cromatografia líquida acoplada à espectrometria de massas (LC-MS). Os resultados obtidos, uma vez que não possuíam distribuição normal, foram submetidos ao teste de Mann Whitney. Foi observada maior concentração de CO exalado, maior concentração salivar de cotinina, maior pontuação no AUDIT, menor viscosidade salivar e resultado desfavorável na oximetria. Em relação ao proteoma salivar, 48 proteínas foram específicas do GCE, sendo que 3 das proteínas desse conjunto estão diferencialmente expressas nesse grupo. Conclui-se que os usuários de cigarro eletrônico apresentam maiores índices de consumo de bebida alcoólica e uma menor viscosidade salivar, alterações importantes na capacidade respiratória demonstrada pelo aumento da concentração de CO no ar expirado e diminuição da oximetria e altos níveis de cotinina presentes na saliva. Além disso, a maior abundância de proteínas, como Glutationa S-transferase P, Cadeia J de imunoglobulina e Acetína desidrogenase, sugerem uma resposta do organismo ao aumento do estresse oxidativo, presença de componentes inflamatórios e alterações microbiológicas salivares que podem favorecer o desenvolvimento de condições patológicas futuras.(AU)

The electronic cigarette, also known as e-cig, has been widely used in recent years, especially by younger populations. Cytotoxic and carcinogenic substances may be present in its composition; however, little is known about the risks of its use. The aim of this study was to evaluate the protein profile of saliva from electronic cigarette users through proteomic analysis and the possible salivary alterations in these individuals. For this purpose, participants were divided into two groups: the Electronic Cigarette Group (ECG), composed of 25 regular and exclusive e-cig users, and the Control Group (CG), consisting of 25 non-smokers and non-e-cig users, matched by sex and age to the ECG. All participants underwent clinical examination and unstimulated saliva collection for salivary flow rate measurement and evaluation of salivary proteome. The samples were aliquoted and subjected to the following analyses: salivary viscosity, pH, buffer capacity, and measurement of salivary cotinine performed using the enzyme-linked immunosorbent assay (ELISA) Cotinine Enzyme Immunoassay Kit (Salimetrics). The participants' alcohol consumption profile was assessed using the Alcohol Use Disorders Identification Test (AUDIT) and questions related to alcohol consumption. The e-cig consumption profile was traced through questions related to device usage and testing for carbon monoxide concentration in expired air. Proteomic analysis was performed using shotgun methodology and liquid chromatography coupled with mass spectrometry (LC-MS). Since the obtained results did not follow a normal distribution, they were subjected to the Mann-Whitney test. A higher concentration of exhaled CO was observed, along with a higher salivary concentration of cotinine, higher scores on the AUDIT, lower salivary viscosity, and an unfavorable result on oximetry. Regarding the salivary proteome, 48 proteins were specific to GCE, with 3 of these proteins being differentially expressed in this group. It is concluded that electronic cigarette users have higher rates of alcohol consumption and lower salivary viscosity, important alterations in respiratory capacity demonstrated by increased CO concentration in expired air and decreased oximetry, and high levels cotinine present in saliva. Furthermore, the higher abundance of proteins such as Glutathione S-transferase P, Immunoglobulin J chain, and Acetyl-CoA dehydrogenase, suggest an organism response to increased oxidative stress, presence of inflammatory components, and salivary microbiological alterations that may favor the development of future pathological conditions (AU)

Prenatal and Childhood Smoke Exposure Associations with Cognition, Language, and Attention-Deficit/Hyperactivity Disorder.

OBJECTIVE: To assess the relationships of prenatal and childhood smoke exposure with specific neurodevelopmental and behavioral problems during early childhood. STUDY DESIGN: A subsample (n = 386) of mother-child dyads from the Newborn Epigenetic Study (NEST) prebirth cohort participated in the study. Cotinine concentrations were used to objectively measure prenatal and childhood smoke exposure when youth were aged 3-13 years. Multivariable regression models were used to estimate associations of prenatal and childhood cotinine concentrations with performance on the National Institutes of Health (NIH) Toolbox and attention-deficit/hyperactivity disorder and behavioral symptoms, measured using the Behavior Assessment System for Children, 2nd edition (BASC-2). RESULTS: After adjusting for confounders, childhood cotinine concentrations were associated with poorer cognitive performance on tasks measuring cognitive flexibility (B = -1.29; P = .03), episodic memory (B = -0.97; P = .02), receptive language development (B = -0.58; P = .01), and inhibitory control and attention (B = -1.59; P = .006). Although childhood cotinine concentration was associated with higher levels of attention problems (B = 0.83; P = .004) on the BASC-2, after adjustment for confounders, the association is nonsignificant. Although associations for maternal cotinine concentrations were null, an interaction was detected between prenatal and childhood cotinine concentrations on the NIH Toolbox Picture Vocabulary Task (P = .02). CONCLUSIONS: Our findings suggest that childhood tobacco smoke exposure may lead to poorer attention regulation and language acquisition, complex visual processing ability, and attention problems.

Avaliação da influência do tabagismo no desenvolvimento da periodontite apical em ratos wistar / Evaluation of smoking influence on the development of apical periodontitis in wistar rats

A infecção endodôntica ocorre a após contaminação do tecido pulpar levando à uma colonização bacteriana dos canais radiculares resultando em uma resposta inflamatória dos tecidos periapicais e formação de uma lesão periapical, a periodontite apical (PA). O tabagismo, por sua vez, tem impactos prejudiciais à saúde oral e sistêmica sendo considerado um importante fator de risco para as doenças periodontais. Este trabalho tem por finalidade investigar a influência do tabagismo no desenvolvimento da periodontite apical em ratos. Trinta e dois ratos machos Wistar foram divididos em 4 grupos experimentais (n=8): Controle (sem periodontite apical induzida e sem inalação da fumaça do cigarro); FU (com inalação a fumaça do cigarro); PA (com periodontite apical induzida); FU+PA (com inalação a fumaça do cigarro e periodontite apical induzida). Para a inalar a fumaça do cigarro, grupo de cinco animais permaneceram em uma câmara de tabagismo inalando a fumaça de 10 cigarros por 8 minutos, três vezes ao dia, durante 50 dias. Decorridos 20 dias de inalação de fumaça do cigarro, foi realizada a cirurgia para indução da periodontite apical nos animais do grupo PA e FU+PA, no primeiro molar inferior direto, o qual permaneceu aberto na cavidade bucal por 30 dias. Nesses 30 dias subsequentes da indução da PA, os animais do grupo FU+PA e FU continuaram com a inalação a fumaça do cigarro. No 50º dia, os animais foram eutanasiados e coletados amostras de tecido hematológico para avalição dos níveis séricos de nicotina, cotinina, fosfatase alcalina, cálcio, fósforo, série vermelha e série branca. As hemimandibula removidas foram escaneadas em microtomógrafo para avaliar o volume da destruição óssea e processadas histologicamente para avaliação do perfil inflamatório e expressão das citocinas IL-6, IL-1ß, TNF-α e dos marcadores do metabolismo ósseo RANKL e OPG. Os dados foram tabulados e aplicados os testes estatísticos de Mann-Whitney para os dados não paramétricos e teste t para os dados paramétricos, a um nível de significância de P< 0.05. Com relação a análise histológica o grupo FU+PA apresentou infiltrado inflamatório mais intenso em relação aos demais grupos (P< 0,05). As citocinas pró-inflamatórias IL-6, IL-1ß, TNF-α apresentaram alto padrão de imunomarcação para o grupo FU+PA (P< 0,05). A análise histométrica mostrou maior área de reabsorção óssea no grupo FU+PA, assim como na análise microtomográfica (P< 0,05). As citocinas RANKL esteve mais expressa no grupo FU+PA (P< 0,05) e OPG teve maior expressão no grupo PA (P< 0,05). Na análise hematológica houve um aumento na concentração de hemácias, hemoglobina e leucócitos para o FU+PA (P< 0,05). Os níveis séricos de cálcio foram menores no FU+PA (P> 0,05), a fosfatase alcalina e o cálcio se manteve constante em todos os grupos experimentais (P> 0,05). A concentração sérica de nicotina e cotinina no grupo FU e FU+PA foram compatíveis com fumante humano(AU)

Endodontic infection occurs mainly after pulp tissue contamination by a carious process, leading to bacterial colonization of root canal system and inflammatory response of periapical tissues, followed by formation of apical periodontitis (AP). It is widely known that smoking has harmful impacts on oral and systemic health and is considered a risk factor for periodontal diseases. The objective of this research was to investigate the influence of smoking on the development of AP in rats. Thirty-two male Wistar rats were divided into 4 experimental groups (n = 8): C - Control (no induced AP and no cigarette smoke inhalation); CSI (cigarette smoke inhalation); AP (induced apical periodontitis); CSI + AP (cigarette smoke inhalation and induced apical periodontitis). To inhale cigarette smoke, group with five animals remained in a smoking chamber inhaling smoke from 10 cigarettes for 8 minutes, three times daily, for 50 days. After 20 days of cigarette smoke inhalation, pulp chamber access was performed to induce apical periodontitis in the first mandibular right molar, which remained with pulp chamber exposed to oral cavity for 30 days in animals in the AP and CSI + AP group. During these 30 days after the AP induction, animals in the CSI + AP and CSI group continued to inhale cigarette smoke. On the 50th day, animals were euthanized and blood sample collected to assess serum levels of nicotine, cotinine, alkaline phosphatase, calcium, phosphorus, red series and white series. The hemimandibles were removed and scanned in microtomograph to assess bone volume and histologically processed to assess inflammatory profile and expression of cytokines IL-6, IL-1ß, TNF-α and bone metabolism markers RANKL and OPG. Data were tabulated and statistically analyzed using Mann-Whitney tests for nonparametric data and analysis of variation test for parametric data, with a significance level of P< 0.05. Regarding histological analysis, CSI+AP group showed more intense inflammatory infiltrate compared to other groups (P < 0.05). Histometric analysis showed a larger area of bone resorption in the CSI + AP group, also observed in the microtomographic analysis (P< 0.05). Group CSI+AP had elevated pro-inflammatory cytokines IL-6, IL-1ß, TNF-α expression (P< 0.05). The RANKL cytokines were also more expressed in the CSI+AP group (P< 0.05), while OPG was more expressed in the AP group (P< 0.05). The hematological analysis revealed an increase in the concentration of red blood cells, hemoglobin, and leukocytes for CSI+AP (P< 0.05). Serum calcium levels were lower in CSI+AP (P> 0.05), and alkaline phosphatase and calcium remained constant in all experimental groups (P> 0.05). The serum concentrations of nicotine and cotinine in the CSI and CSI+AP group were compatible with human smokers(AU)

Brief Report: HIV Infection Does Not Explain Higher Nicotine Metabolism in People Living With HIV.

BACKGROUND: Smoking contributes to significant morbidity and mortality in people with HIV. People with HIV have relatively high nicotine metabolism rates, as measured by the nicotine metabolite ratio (NMR, 3-hydroxycotinine/cotinine). A higher NMR is associated with difficulty quitting smoking. We hypothesized that HIV infection might upregulate nicotine metabolism. SETTING: A retrospective study of male current smokers in the Multicenter AIDS Cohort Study who HIV seroconverted between 1985 and 1993. METHODS: Eligibility included having plasma stored before and after confirmed HIV seroconversion and current tobacco use. Samples were selected from the closest available visits before (median 3.3 months) and after (median 9.4 months) seroconversion. Antiretroviral therapy use was exclusionary. Cotinine and 3-hydroxycotinine were measured using liquid chromatography-tandem mass spectrometry. We compared NMR from plasma pre-HIV and post-HIV infection using signed-rank tests. We targeted a sample size of 71 pairs to achieve 80% power to detect a 0.1 unit increase in NMR with P = 0.05. RESULTS: We analyzed paired samples from 78 participants; the median age was 34.5 years [interquartile range (IQR 29-40 years)]. The median NMR pre-HIV and post-HIV was 0.45 (IQR 0.32-0.54) and 0.46 (IQR 0.34-0.56), respectively. The median change in NMR postseroconversion was +0.01 (IQR -0.05, +0.09), P = 0.25. Stratification of median change in NMR by timing between samples or time since HIV seroconversion did not alter this finding. CONCLUSIONS: Acquiring HIV had no measurable effect on NMR. We postulate that upregulation of the NMR may be due to direct pharmacologic effects of HIV medications or metabolic changes in response to HIV infection.

Exposure in the tobacco fields: Genetic damage and oxidative stress in tobacco farmers occupationally exposed during harvest and grading seasons.

Agricultural workers engaged in tobacco cultivation are constantly exposed to large amounts of harmful agents, such as pesticides and nicotine. Furthermore, most of the flue-cured tobacco leaves are manually graded exposing workers to agents such as tobacco-specific nitrosamines. This study aimed to evaluate genetic damage and oxidative stress in tobacco farmers occupationally exposed during the harvest and grading seasons. We obtained data on DNA damage detected in Comet assay in blood cells and micronucleus experiment with buccal cells from 241 individuals. The serum cotinine levels and nitrates were also evaluated. The Comet Assay results showed a showed an increased visual score for males and females during harvest time and tobacco grading. An increase of micronucleated and binucleated cells was observed in the grading group compared to the control and harvest groups. The oxidative stress measurements showed a clear increase of thiobarbituric acid reactive substances (TBARS) in tobacco farmers during harvest time, and trolox equivalent antioxidant capacity (TEAC) in individuals during harvest and grading time compared to the controls. Significant increases of the cotinine levels were observed during the harvest and grading period (harvest>grading), and nitrates for the grading period compared to the control. In this study, tobacco farmers presented compromised DNA integrity associated with enhanced oxidative stress levels.

Multilevel Intervention for Low-Income Maternal Smokers in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC).

Objectives. To test the efficacy of Babies Living Safe and Smokefree (BLiSS), a multilevel intervention initiated in a citywide safety net health system to improve low-income maternal smokers' abstinence and reduce child tobacco smoke exposure. Methods. This randomized controlled trial in Philadelphia, Pennsylvania (2015-2020), recruited low-income maternal smokers who received a brief smoking intervention (Ask, Advise, Refer [AAR]) from nutrition professionals in the Special Supplemental Nutrition Program for Women, Infants, and Children before randomization to (1) a multilevel intervention (AAR + multimodal behavioral intervention [MBI]; n = 199) or (2) an attention control intervention (AAR + control; n = 197). Results. AAR + MBI mothers had significantly higher 12-month bioverified abstinence rates than did AAR + control mothers (odds ratio [OR] = 9.55; 95% confidence interval [CI] = 1.54, 59.30; P = .015). There were significant effects of time (b = -0.15; SE = 0.04; P < .001) and condition by time (b = -0.19; SE = 0.06; P < .001) on reported child exposure favoring AAR + MBI, but no group difference in child cotinine. Presence of other residential smokers was related to higher exposure. Higher baseline nicotine dependence was related to higher child exposure and lower abstinence likelihood at follow-up. Conclusions. The multilevel BLiSS intervention was acceptable and efficacious in a population that experiences elevated challenges with cessation. Public Health Implications. BLiSS is a translatable intervention model that can successfully improve efforts to address the persistent tobacco-related burdens in low-income communities. Trial Registration. Clinical Trials.gov identifier: NCT02602288. (Am J Public Health. 2022;112(3):472-481. https://doi.org/10.2105/AJPH.2021.306601).

Low-cost apparatus for cigarette smoke exposure in rats.

BACKGROUND: The effects of tobacco smoke on the central nervous system are usually studied with isolated nicotine, ignoring other compounds present in cigarette smoke. The few studies that use in vivo whole-body cigarette smoke exposure are usually performed in expensive commercial apparatus. NEW METHOD: We presented a feasible, safe, and low-cost apparatus for cigarette smoke exposure in rodents. RESULTS: Rats exposed to cigarette smoke in this apparatus showed cotinine levels similar to human active smokers. Additional results showed that cigarette smoke exposure increased glutamate and aspartic acid levels and decreased leucine, isoleucine, ornithine, phenylalanine, and tryptophan levels in the cerebrospinal fluid of rats. COMPARISON WITH EXISTING METHOD(S): Our apparatus is feasible, safe, and costs 67-fold less than a commercial automatized smoking machine. Beyond the low cost, it does not require specialized knowledge for building or maintenance. CONCLUSIONS: We concluded that our low-cost apparatus is reliable and reproduces cigarette smoke use in humans.

Cross-sectional study examining the accuracy of self-reported smoking status as compared to urinary cotinine levels among workers at risk for chronic kidney disease of unknown origin in Guatemala.

OBJECTIVES: There is a lack of information on cotinine levels in rural populations in low-income and middle-income countries like Guatemala. Therefore, there is a need to explore smoking status and biomarkers of tobacco use in epidemiological research in rural, low-income populations, in particular those at-risk for chronic kidney disease of unknown origin (CKDu). DESIGN: We evaluated self-reported smoking status against urinary cotinine levels, the gold standard biomarker of tobacco smoke exposure, among agricultural workers at four separate cross-sectional time points. SETTING: Guatemala. PARTICIPANTS: 283 sugarcane workers. PRIMARY OUTCOME MEASURES: Compared self-reported smoking status and urinary cotinine levels in two agricultural worker studies. RESULTS: Self-reported smoking prevalence was 12% among workers. According to cotinine levels (≥50 ng/mL), the smoking prevalence was 34%. Self-reported smoking status had 28% sensitivity and 96% specificity. Urinary cotinine levels show that smoking prevalence is underestimated in this worker population. CONCLUSIONS: According to our findings, smoking status should be objectively measured with biomarkers rather than self-reported in CKDu epidemiological research. Self-reported smoking status is likely an underestimate of the true smoking prevalence among agricultural workers. Research on the CKDu epidemic in Central America and other parts of the world might be underestimating tobacco exposure as a potential contributor to the development of CKDu.

Socio-demographic and psychological features associated with smoking in pregnancy.

OBJECTIVE: To investigate how social and psychological characteristics differ between pregnant women who smoke and do not smoke. To explore associations between social and psychological features with changes of smoking habits by the end of pregnancy. METHODS: A case-control study was set up. Smokers cases were never-smokers and ex-smokers controls. Pregnant women (n=328) from public prenatal services were interviewed. Socio-demographic data and psychological variables - personality traits, anxiety, depression, perceived stress, maternal fetal-attachment - were measured. Saliva samples were collected to measure cotinine and to check self-informed smoking status. In addition, 66 smokers were also assessed regarding smoking habits by late pregnancy. Smoking status was defined as a dependent variable. Exposure factors were analyzed through odds ratios. Logistic models and contingency tables were employed according to the nature of variables. "Qualitative change in smoking" was defined as a dependent variable for the last evaluation, and a logistic regression model was built. RESULTS: Lower schooling, higher age, use of alcohol and drugs, living without a partner, and passive smoking showed associations with smoking. Anxiety, depression and perceived stress also exhibited positive association with smoking. Among personality traits, only Neuroticism was associated with smoking. None of the variables were associated with qualitative change in smoking by the end of pregnancy. CONCLUSION: Smoking during pregnancy is associated with more unfavorable social conditions. Pregnant women who smoke exhibit more negative psychological states than nonsmokers, including a profile of accentuated Neuroticism. None of the investigated variables could predict changes in smoking during pregnancy.

Short-term effects of nicotine gum on facial detection in healthy nonsmokers: a pilot randomized controlled trial.

Objective: The main purpose of this study was to investigate short-term effects of nicotine gum on facial detection. Methods: Fourteen participants (mean age = 26.8 years, SD = 2.5 years; eight males) were enrolled in this pilot randomized controlled trial of nicotine gum administration (placebo, 2-mg and 4-mg doses). The participants were instructed to detect the location of a face when it was presented in a face/nonface pair on the screen. A repeated multivariate analysis of variance was conducted to analyze the results for reaction time and discrimination index. Demographics were used to explore significant association on facial detection. Bayesian analyses were also carried out considering maximum robustness to avoid bias. Results: The results indicated that the 2-mg dose resulted in faster reaction time and better discrimination than the 4-mg dose (p < 0.001). The 4-mg dose resulted in slower reaction time and lower discrimination index compared to both placebo (p < 0.01) and 2-mg doses (p < 0.001). Demographic data were not related to the outcomes. Conclusions: The results indicate that nicotine improved facial detection, but only at low doses (i.e., 2-mg), following a U-shaped curve. We trust future studies will continue to advance this research field, and if further work supports these preliminary findings, nicotine can act as therapeutic target in populations such as those with low vision.

The role of glucose metabolism and insulin resistance in cardiac remodelling induced by cigarette smoke exposure.

The aim of this study is to evaluate whether the alterations in glucose metabolism and insulin resistance are mechanisms presented in cardiac remodelling induced by the toxicity of cigarette smoke. Male Wistar rats were assigned to the control group (C; n = 12) and the cigarette smoke-exposed group (exposed to cigarette smoke over 2 months) (CS; n = 12). Transthoracic echocardiography, blood pressure assessment, serum biochemical analyses for catecholamines and cotinine, energy metabolism enzymes activities assay; HOMA index (homeostatic model assessment); immunohistochemistry; and Western blot for proteins involved in energy metabolism were performed. The CS group presented concentric hypertrophy, systolic and diastolic dysfunction, and higher oxidative stress. It was observed changes in energy metabolism, characterized by a higher HOMA index, lower concentration of GLUT4 (glucose transporter 4) and lower 3-hydroxyl-CoA dehydrogenase activity, suggesting the presence of insulin resistance. Yet, the cardiac glycogen was depleted, phosphofructokinase (PFK) and lactate dehydrogenase (LDH) increased, with normal pyruvate dehydrogenase (PDH) activity. The activity of citrate synthase, mitochondrial complexes and ATP synthase (adenosine triphosphate synthase) decreased and the expression of Sirtuin 1 (SIRT1) increased. In conclusion, exposure to cigarette smoke induces cardiac remodelling and dysfunction. The mitochondrial dysfunction and heart damage induced by cigarette smoke exposure are associated with insulin resistance and glucose metabolism changes.

Fetal and Postnatal Nicotine Exposure Modifies Maturation of Gonocytes to Spermatogonia in Mice.

Studies in laboratory animals have shown that male offspring from dams, exposed to nicotine during pregnancy and postnatal periods, show alterations in fertility, although the origin of this is still uncertain. In this study, we examined in a mouse model if the process of gonocyte maturation to spermatogonia was affected in male offspring from dams with nicotine administration during pregnancy and postnatal periods. BALB/C mice, with and without nicotine administrations in pregnancy and postnatal periods, were studied. The animals were euthanized at 3, 7, 10, 16, and 35 days postpartum (dpp). Testicular tissue samples were processed for histological, ultrastructural, and immunohistochemical studies; and testicular lipoperoxidation was determined. It was observed that in the nicotine-exposed animals, there was increased apoptosis and a reduction in the number of gonocytes that matured to spermatogonia. This gonocyte-spermatogonia maturation reduction was associated with a greater immunoreactivity to nicotinic acetylcholine receptors in the germ cells. Lipoperoxidation was similar in both groups until 16 dpp, with significant reduction at 35 dpp. Our findings suggest that nicotine intake during pregnancy and postnatal periods can affect the process of maturation of gonocytes to spermatogonia and the pool of available spermatogonia for spermatogenesis.

Impact of nicotine-induced green tobacco sickness on DNA damage and the relation with symptoms and alterations of redox status in tobacco farmers.

During the harvest period, tobacco workers are exposed to nicotine and it is known that absorption of the alkaloid via the leaves causes green tobacco sickness (GST). We investigated if GST and its symptoms are associated with DNA damage and alterations of the redox status. DNA damage was measured in lymphocytes of tobacco workers and controls (n = 40/group) in single cell gel electrophoresis assays. Exposure to nicotine was determined by plasma cotinine measurements, alterations of the redox status by quantification of the total antioxidant capacity (TEAC) and of thiobarbituric acid reactive substances (TBARS). The symptoms of GTS included nausea, abdominal cramps, headache, vomiting and dizziness, and 50% of the workers had more than one symptom. Cotinine levels were enhanced in the workers (111 ng/mL); furthermore, the extent of DNA damage was ca. 3-fold higher than in the controls. This effect was more pronounced in participants with GST compared to healthy nicotine exposed workers and increased in individuals with specific symptoms (range 22-36%). TBARS levels did not differ between workers and unexposed controls, while TEAC values were even increased (by 14.3%). Contact with nicotine present in tobacco leaves causes GTS and leads to damage of the DNA; this effect is more pronounced in workers with GTS symptoms and is associated with alterations of the redox status. Damage of the genetic material which was found in the workers may lead to adverse long-term effects that are caused by genomic instability such as cancer and accelerated ageing.

Prevalence of urinary cotinine levels in children under 5 years of age during consultations for acute respiratory disease at the emergency department of the Universidad de La Sabana clinic.

BACKGROUND: Several environmental factors favour the occurrence of acute respiratory disease, which is the main reason for paediatric consultations in our country (Colombia). Tobacco smoke is considered a significant environmental pollutant with a great impact on health. The objective of this study is to estimate the prevalence of cotinine levels measured in urine, in children between 1 to 60 months of age who attended an emergency department with acute respiratory disease. METHODS: A cross-sectional study was conducted that included children between 1 and 60 months of age with acute respiratory disease who were admitted to the emergency department of the Universidad de La Sabana Clinic between April and July 2016. RESULTS: We included 268 patients and 36% were female. Of the total population examined, 33.96% showed positive results for urinary cotinine, of whom 97.8% had values between 10 and 100 ng/ml, which is considered positive for exposure to second-hand smoke. The principal pathology was recurrent wheezing in 43.96% of these cases. Regarding the presence of smokers at home, it is important to mention that in 54.95% of the children with positive urinary cotinine test was no related with smokers at home. And in 45.05% of positive urinary cotinine was evidence of smokers at home, being associated with the positive result P <  0.001 and smoking within the house P = 0.018; smoking when children were present did not have significant P = 0.105. The activities performed after smoking such as hand washing, change of clothes, eating, brushing teeth, did not influence the test result P = 0.627. CONCLUSIONS: A high prevalence of urinary cotinine was observed, which is associated with the presence of a smoker at home, and this relationship was independent of the activities performed by the smoker after smoking. In addition, a positive test for urinary cotinine was presented in some children without documented exposure to cigarette smoke inside the home, which may be explained by the presence of environmental cotinine. Therefore, it is necessary to perform educational interventions aimed at parents and caregivers who smoke.

Pregnant Smokers Receiving Opioid Agonist Therapy Have an Elevated Nicotine Metabolite Ratio: A Replication Study.

INTRODUCTION: Pregnant women exposed chronically to opioids smoked more cigarettes per day (CPD) and had a higher nicotine metabolite ratio (NMR), 3-hydroxycotinine/cotinine, a biomarker of nicotine metabolism and clearance, than those not receiving opioids. We examined CPD and NMR in a group of pregnant smokers, a quarter of whom were receiving opioid agonist therapy (OAT). AIMS AND METHODS: Pregnant smokers recruited to participate in a placebo-controlled trial of bupropion for smoking cessation provided a blood sample for measurement of NMR. RESULTS: Half (52.4%) of the 124 women with NMR data were African American. OAT-treated women (n = 34, 27.4%; 27 receiving methadone and 7 buprenorphine) were more likely to be white (79% vs. 30%, p < .001) and to have a lower mean PHQ-9 total score (2.91 [SD = 2.83] vs. 4.83 [SD = 3.82], p = .007). OAT-treated women reported smoking more CPD (9.50 [SD = 5.26] vs. 7.20 [SD = 3.65], p = .005) and had higher NMR (0.78 [SD = 0.36] vs. 0.56 [SD = 0.25], p = .001) than the non-OAT-treated group. In a linear regression analysis adjusting for race, depression severity, and CPD, NMR was greater in the OAT group (p = .025), among whom the daily methadone-equivalent dosage correlated with NMR (Spearman's ρ = 0.49, p = .003). CONCLUSIONS: Consistent with the findings of Oncken et al. (2019), we found that OAT smokers smoked more and had higher NMR than non-OAT smokers. As higher NMR is associated with a reduced likelihood of smoking cessation, the effects on NMR of both pregnancy and OAT could contribute to a lower smoking cessation rate in pregnant smokers receiving chronic opioid therapy. IMPLICATIONS: We replicated the finding that the NMR is significantly greater among pregnant smokers receiving OAT than those not receiving this treatment for opioid use disorder. Furthermore, we found that the dosage of the OAT was significantly associated with the NMR level. These findings may contribute to a poorer response to smoking cessation treatment in pregnant women treated with OAT, particularly those receiving high-dose therapy, and raise the question of whether novel approaches are needed to treat smoking in this subgroup of pregnant smokers.

Early life tobacco exposure and children's telomere length: The HELIX project.

Telomere length and mitochondrial DNA content are considered biomarkers of cellular aging, oxidative stress, and inflammation, but there is almost no information on their association with tobacco smoke exposure in fetal and early life. The aim of this study was to assess whether prenatal and childhood tobacco exposure were associated with leukocyte telomere length (LTL) and mitochondrial DNA (mtDNA) content in children. As part of a multi-centre European birth cohort study HELIX (Human Early-Life Exposome) (n = 1396) we assessed maternal smoking status during pregnancy through questionnaires, and through urinary cotinine levels that were then used to classify women as not exposed to smoking (<10 µg/L), exposed to secondhand smoke (SHS) (10-50 µg/L) and active smokers (>50 µg/L). When the children were around 8 years of age (range: 5.4-12.0 years), childhood SHS tobacco smoke exposure was assessed through an extensive questionnaire and through measurements of urinary cotinine (<3.03 µg/L non-detected, >3.03 µg/L detected). Leukocyte mtDNA content and LTL were measured in the children at 8 years employing real time polymerase chain reaction (qPCR). Effect estimates were calculated using multivariate linear regression models for prenatal and childhood exposures adjusted for potential confounders. Maternal cotinine levels indicative of SHS exposure during pregnancy were associated with a decrease of 3.90% in LTL in children (95% CI: -6.68, -0.91), compared with non-smoking, whereas the association for maternal cotinine levels indicative of active smoking did not reach statistical significance (-3.24%; 95% CI: -6.59, 0.21). Childhood SHS tobacco exposure was not associated with LTL in children. Global SHS exposure during childhood was associated with an increase of 3.51% (95% CI: 0.78, 6.27) in mtDNA content. Our findings suggest that tobacco smoke exposure during pregnancy, even at SHS levels, may accelerate telomere shortening in children and thus induce biological aging from an early age.

Associations Between Early Low-Level Tobacco Smoke Exposure and Executive Function at Age 8 Years.

OBJECTIVE: To investigate whether exposure to tobacco smoke during early brain development is linked with later problems in behavior and executive function. STUDY DESIGN: We studied 239 children in a prospective birth cohort. We measured tobacco exposures by caregiver report and serum cotinine 3 times during pregnancy and 4 times during childhood. We used linear regression to examine the association between prenatal and childhood serum cotinine concentrations and behavior (the Behavior Assessment System for Children-2) and executive function (the Behavior Rating Inventory of Executive Function) at age 8 years while adjusting for important covariates. RESULTS: Neither prenatal nor child serum cotinine were associated with behavior problems measured by the Behavior Assessment System for Children-2. On the Behavior Rating Inventory of Executive Function, prenatal and childhood exposure was associated with poorer task initiation scores (B = 0.44; 95% CI, 0.03-0.85 and B = 0.69, 95% CI, 0.06-1.32 respectively). Additionally, in a subset of 208 children with nonsmoking mothers, prenatal exposure was associated with task initiation scores (B = 1.17; 95% CI, 0.47-1.87) and additional components of the metacognition index (eg, working memory, B = 1.20; 95% CI, 0.34-2.06), but not components of the behavioral regulation index. CONCLUSIONS: Tobacco exposures during pregnancy (including low-level second-hand smoke) and childhood were associated with deficits in some domains of children's executive function, especially task initiation and metacognition. These results highlight that decreasing early exposure to tobacco smoke, even second-hand exposure, may support ideal brain functioning.

Parents' Self-efficacy for Tobacco Exposure Protection and Smoking Abstinence Mediate Treatment Effects on Child Cotinine at 12-Month Follow-up: Mediation Results from the Kids Safe and Smokefree Trial.

INTRODUCTION: Compared with the general smoking population, low-income smokers face elevated challenges to success in evidence-based smoking cessation treatment. Moreover, their children bear increased disease burden. Understanding behavioral mechanisms related to successful reduction of child tobacco smoke exposure (TSE) could inform future smoking interventions in vulnerable, underserved populations. METHODS: Smoking parents were recruited from pediatric clinics in low-income communities and randomized into a multilevel intervention including a pediatric clinic intervention framed in best clinical practice guidelines ("Ask, Advise, Refer" [AAR]) plus individualized telephone counseling (AAR + counseling), or AAR + control. Mediation analysis included treatment condition (independent variable), 12-month child cotinine (TSE biomarker, criterion), and four mediators: 3-month end-of-treatment self-efficacy to protect children from TSE and smoking urge coping skills, and 12-month perceived program (intra-treatment) support and bioverified smoking abstinence. Analyses controlled for baseline nicotine dependence, depressive symptoms, child age, and presence of other residential smokers. RESULTS: Participants (n = 327) included 83% women and 83% African Americans. Multilevel AAR + counseling was associated with significantly higher levels of all four mediators (ps < .05). Baseline nicotine dependence (p < .05), 3-month self-efficacy (p < .05) and 12-month bioverified smoking abstinence (p < .001) related significantly to 12-month child cotinine outcome. The indirect effects of AAR + counseling intervention on cotinine via self-efficacy for child TSE protection and smoking abstinence (ps < .05) suggested mediation through these pathways. CONCLUSIONS: Compared with AAR + control, multilevel AAR + counseling improved all putative mediators. Findings suggest that fostering TSE protection self-efficacy during intervention and encouraging parental smoking abstinence may be key to promoting long-term child TSE-reduction in populations of smokers with elevated challenges to quitting smoking. IMPLICATIONS: Pediatric harm reduction interventions to protect children of smokers from tobacco smoke have emerged to address tobacco-related health disparities in underserved populations. Low-income smokers experience greater tobacco-related disease burden and more difficulty with smoking behavior change in standard evidence-based interventions than the general population of smokers. Therefore, improving knowledge about putative behavioral mechanisms of smoking behavior change that results in lower child exposure risk could inform future intervention improvements.

Evaluation of oral mechanical and gustatory sensitivities and salivary cotinine levels in adult smokers.

Objective: The aim was to examine oral mechanical and gustatory sensitivities in adult smokers and to estimate salivary levels of cotinine by tobacco consumption. A total of 54 adults (20-45 years old; 28 males/26 females) were divided into two sex-paired groups: smoker group (n = 27), tobacco consumers with no other chronic disease/use of chronic medication, and a control non-smoker non-exposed group with similar age (n = 27).Materials and Methods: 24 h-Recall was used to gather information about tobacco consumption, date of onset and duration of the habit. Oral mechanical evaluation comprised touch detection threshold (MDT) of upper and lower lips and tongue tip and two-point discrimination (TPD) assessments. Taste sensitivities for sweet, salty, sour and bitter were evaluated in four concentrations. Salivary cotinine was determined by high performance liquid chromatography. Statistical analysis comprised Mann-Whitney, Two-way ANOVA test and regression analysis.Results: The mean smoking time was 13.6 years (mean 8.4 mg/day; 13 cigarettes/day). A sex-effect was observed on MDT of tongue tip (higher sensitivity in females), while group-effect was observed on TPD of lower lip, showing a smaller sensitivity among smokers (p < .05; moderate effect: Eta partial2 = 0.076). Although the total score of gustatory sensitivity did not differ between groups, smokers exhibited an irregular pattern of correctly identified tastants among the different concentrations of salty, sour and bitter. The predictive model showed that salivary cotinine was dependent on "nicotine consumption on the day before" (R2 = 49%).Conclusion: A difference in tactile sensitivity of the lower lip and qualitative changes in taste sensitivity were observed in smokers.

Cigarette smoking and antioxidant defences in packed red blood cells prior to storage.

BACKGROUND: Red blood cells from smoking donors can have more lesions from oxidative stress, decreasing the benefits of blood transfusion. We aimed to explore the effect of cigarette smoking on the oxidative status of packed red blood cells (PRBCs) prior to storage. MATERIALS AND METHODS: We compared serum vitamin C, plasmatic malondialdehyde (MDA), and non-protein thiol groups (GSH) levels in PRBCs, as well glutathione peroxidase (GPx) and glutathione s-transferase (GST) activity in PRBCs from smoking (n=36) and non-smoking (n=36) donors. We also correlated urinary cotinine levels with these parameters. RESULTS: Cigarette smoking was associated with decreased serum levels of vitamin C and GPx, and increased GST activity in PRBCs. We found negative correlations between cotinine, GPx activity and vitamin C levels, and a positive correlation between cotinine and GST activity. DISCUSSION: Cigarette smoking changed antioxidant defences of PRBCs prior to storage and these parameters are correlated with cotinine levels. Increased RBC antioxidants such as GST may reflect an exposure to oxidants during erythropoiesis. Because of the inability of mature RBCs to resynthesise antioxidants, PRBCs from smokers may have higher risk of storage lesions than those from non-smoker donors.