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)

Nicotinic acetylcholine receptors and learning and memory deficits in Neuroinflammatory diseases.

Animal survival depends on cognitive abilities such as learning and memory to adapt to environmental changes. Memory functions require an enhanced activity and connectivity of a particular arrangement of engram neurons, supported by the concerted action of neurons, glia, and vascular cells. The deterioration of the cholinergic system is a common occurrence in neurological conditions exacerbated by aging such as traumatic brain injury (TBI), posttraumatic stress disorder (PTSD), Alzheimer's disease (AD), and Parkinson's disease (PD). Cotinine is a cholinergic modulator with neuroprotective, antidepressant, anti-inflammatory, antioxidant, and memory-enhancing effects. Current evidence suggests Cotinine's beneficial effects on cognition results from the positive modulation of the α7-nicotinic acetylcholine receptors (nAChRs) and the inhibition of the toll-like receptors (TLRs). The α7nAChR affects brain functions by modulating the function of neurons, glia, endothelial, immune, and dendritic cells and regulates inhibitory and excitatory neurotransmission throughout the GABA interneurons. In addition, Cotinine acting on the α7 nAChRs and TLR reduces neuroinflammation by inhibiting the release of pro-inflammatory cytokines by the immune cells. Also, α7nAChRs stimulate signaling pathways supporting structural, biochemical, electrochemical, and cellular changes in the Central nervous system during the cognitive processes, including Neurogenesis. Here, the mechanisms of memory formation as well as potential mechanisms of action of Cotinine on memory preservation in aging and neurological diseases are discussed.

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.

Profile of tobacco growers with green tobacco sickness in Southern Brazil.

Introduction: Green tobacco sickness affects tobacco growers while handling tobacco leaves, regardless of the cultivation stage. Objectives: To characterize the sociodemographic and occupational profiles of tobacco growers with green tobacco sickness, as well as their health habits. Methods: This was a cross-sectional, descriptive study with information from a database obtained from a previous study conducted in a Southern Brazilian municipality. The data were analyzed using absolute and relative frequency. Results: We identified 8 cases of tobacco growers with green tobacco sickness, whose sociodemographic and occupational profiles and health habits were described. Conclusions: The data obtained in this study corroborate the existing literature on tobacco growers. The sociodemographic and occupational profiles and the health habits of our study participants have already been described in other studies, as well as of tobacco growers without green tobacco sickness.

Is secondhand smoke exposure associated with poor periodontal status in children and adolescents? A systematic review and meta-analysis.

PURPOSE: We aimed to systematically evaluate whether SHS exposure is associated with poor periodontal status in individuals up to 15 years. METHODS: Seven databases were searched by two independent reviewers according to pre-specified eligibility criteria up to November 2021. The methodological quality of included studies was appraised using The Newcastle-Ottawa Scale and GRADE was used for assessing the certainty of evidence. Random-effects pairwise meta-analyses compared the periodontal status of those exposed and unexposed to SHS through standardized mean differences (SMDs) and associated confidence intervals (95% CI). RESULTS: Eight cross-sectional studies were eligible for inclusion and two present high methodological quality. All studies contributed to the meta-analysis for gingival index scores (GI) and four for probing pocket depth (PPD). Those exposed exhibited significantly higher levels of GI compared to unexposed (SMD = 1.03, 95% CI 0.17-1.89), but no difference was observed for PPD (SMD = 0.34, 95% CI - 0.14-0.82), with overall very low certainty on evidence. CONCLUSION: Therefore, very low certainty evidence supports that children and adolescents exposed to SHS possibly present poorer periodontal status due to higher levels of GI.

Child Labor in Family Tobacco Farms in Southern Brazil: Occupational Exposure and Related Health Problems.

Tobacco farming is considered Hazardous Child Labor in Brazil. This study examined the work of children and adolescents in tobacco farming, characterizing the level of urinary cotinine and the occurrence of Green Tobacco Sickness (GTS), pesticide poisoning, respiratory symptoms, and musculoskeletal disorders. A cross-sectional descriptive study was conducted with a random sample of tobacco growers under 18 years old in Southern Brazil. Ninety-nine young people were interviewed at 79 family farms. The majority began working in agriculture before they were 14 and worked harvesting and tying hands of tobacco; 60% were 16 or 17 years old, and 51.5% were male. During their lifetime, 24.5% reported GTS, and 3% reported pesticide poisoning. In the previous year, 29.3% reported low back pain, 6.1% wheezing, and 16.2% coughing without having a cold. Half of the 12 young people evaluated had over 100 ng/mL of urinary cotinine. The study indicates that child laborers do various activities and present a high prevalence of health problems. Health workers should be trained to identify child laborers and their impacts on health. Full-time farm schools could provide knowledge about sustainable agricultural production, reducing the rates of age-grade mismatch, without taking young people away from rural areas.

Cholinergic Receptor Modulation as a Target for Preventing Dementia in Parkinson's Disease.

Parkinson's disease (PD) is a neurodegenerative condition characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) in the midbrain resulting in progressive impairment in cognitive and motor abilities. The physiological and molecular mechanisms triggering dopaminergic neuronal loss are not entirely defined. PD occurrence is associated with various genetic and environmental factors causing inflammation and mitochondrial dysfunction in the brain, leading to oxidative stress, proteinopathy, and reduced viability of dopaminergic neurons. Oxidative stress affects the conformation and function of ions, proteins, and lipids, provoking mitochondrial DNA (mtDNA) mutation and dysfunction. The disruption of protein homeostasis induces the aggregation of alpha-synuclein (α-SYN) and parkin and a deficit in proteasome degradation. Also, oxidative stress affects dopamine release by activating ATP-sensitive potassium channels. The cholinergic system is essential in modulating the striatal cells regulating cognitive and motor functions. Several muscarinic acetylcholine receptors (mAChR) and nicotinic acetylcholine receptors (nAChRs) are expressed in the striatum. The nAChRs signaling reduces neuroinflammation and facilitates neuronal survival, neurotransmitter release, and synaptic plasticity. Since there is a deficit in the nAChRs in PD, inhibiting nAChRs loss in the striatum may help prevent dopaminergic neurons loss in the striatum and its pathological consequences. The nAChRs can also stimulate other brain cells supporting cognitive and motor functions. This review discusses the cholinergic system as a therapeutic target of cotinine to prevent cognitive symptoms and transition to dementia in PD.

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.

Strategies for the Treatment of Parkinson's Disease: Beyond Dopamine.

Parkinson's disease (PD) is the second-leading cause of dementia and is characterized by a progressive loss of dopaminergic neurons in the substantia nigra alongside the presence of intraneuronal α-synuclein-positive inclusions. Therapies to date have been directed to the restoration of the dopaminergic system, and the prevention of dopaminergic neuronal cell death in the midbrain. This review discusses the physiological mechanisms involved in PD as well as new and prospective therapies for the disease. The current data suggest that prevention or early treatment of PD may be the most effective therapeutic strategy. New advances in the understanding of the underlying mechanisms of PD predict the development of more personalized and integral therapies in the years to come. Thus, the development of more reliable biomarkers at asymptomatic stages of the disease, and the use of genetic profiling of patients will surely permit a more effective treatment of PD.

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.

Prenatal Exposure to Tobacco and Offspring Neurocognitive Development in the Healthy Start Study.

OBJECTIVE: To explore the associations between prenatal exposure to tobacco and neurocognitive development, in the absence of prematurity or low birth weight. STUDY DESIGN: We followed mother-child pairs within Healthy Start through 6 years of age. Children were born at ≥37 weeks of gestation with a birth weight of ≥2500 g. Parents completed the Third Edition Ages and Stages Questionnaire (n = 246) and children completed a subset of the National Institutes of Health Toolbox Cognition Battery (n = 200). The Ages and Stages Questionnaire domains were dichotomized as fail/monitor and pass. Maternal urinary cotinine was measured at approximately 27 weeks of gestation. Separate logistic regression models estimated associations between prenatal exposure to tobacco (cotinine below vs above the limit of detection) and the Ages and Stages Questionnaire domains. Separate linear regression models estimated associations between prenatal exposure to tobacco and fully corrected T-scores for inhibitory control, cognitive flexibility, and receptive language, as assessed by the National Institutes of Health Toolbox. A priori covariates included sex, maternal age, maternal education, daily caloric intake during pregnancy, race/ethnicity, household income, maternal psychiatric disorders, and, in secondary models, postnatal exposure to tobacco. RESULTS: Compared with unexposed offspring, exposed offspring were more likely to receive a fail/monitor score for fine motor skills (OR, 3.9; 95% CI, 1.5-10.3) and decreased inhibitory control (B: -3.0; 95% CI, -6.1 to -0.7). After adjusting for postnatal exposure, only the association with fine motor skills persisted. CONCLUSIONS: Prenatal and postnatal exposures to tobacco may influence neurocognitive development, in the absence of preterm delivery or low birth weight. Increased developmental screening may be warranted for exposed children.

Occurrence of green tobacco sickness and associated factors in farmers residing in Dom Feliciano Municipality, Rio Grande do Sul State, Southern Region of Brazil / Ocorrência da doença da folha verde do tabaco e fatores associados entre fumicultores no Município de Dom Feliciano, Rio Grande do Sul, Região Sul do Brasil / Ocurrencia de la enfermedad del tabaco verde y factores asociados en agricultores que residen en el Municipio de Dom Feliciano, Estado de Rio Grande do Sul, Región Sur de Brasil

Abstract: This study aimed to evaluate the occurrence of the green tobacco sickness (GTS) and its associated factors in tobacco familiar farmers residing in Dom Feliciano, Rio Grande do Sul State, Brazil. A cross-sectional study was conducted evaluating the sickness in 354 small tobacco farmers, between October 2011 and March 2012. The urinary concentration of cotinine, a biomarker of exposure to nicotine, was determined during the tobacco harvest period. Subjects presenting cotinine urinary levels ≥ 50ng/mL, that had contact with tobacco leaves up to 48 hours before the sample collection and reported at least one disease symptom were deemed as cases. A non-conditional logistic analysis was performed to evaluate the association between GTS and the population characteristics. A total of 122 (34.5%) cases were identified, with 39% of them being smokers and 61% being males. The median cotinine urinary concentrations were 75.6ng/mL (74.1ng/mg of creatinine) for non-cases and 755.8ng/mL (632.1ng/mg of creatinine) for the cases (p-value ≤ 0.01). The multivariate analysis showed a positive association between GTS and sunlight exposure time, exposure to pesticides, worse health status, and inverse association with wood cultivation. This study presented a high GTS prevalence and suggest that the use of urinary cotinine is a significant biomarker to determine GTS cases, influencing in the distribution by sex. Once the tobacco production involves health hazards, is important to implement measures to prevent the harm caused to tobacco farmers, as set in articles 17 and 18 of the World Health Organization Framework Convention on Tobacco Control.

Resumo: O estudo teve como objetivo avaliar a ocorrência de doença da folha verde (DFV) e fatores associados entre fumicultores familiares no Município de Dom Feliciano, Estado do Rio Grande do Sul, Brasil. Foi realizado um estudo transversal sobre DFV em 354 pequenos produtores de tabaco, entre outubro de 2011 e março de 2012. Foi medida a concentração urinária da cotinina, um biomarcador da exposição à nicotina, durante a temporada de colheita do tabaco. Os casos de DFV foram definidos como indivíduos com níveis urinários de cotinina ≥ 50ng/mL, que tiveram contato com folhas de tabaco em até 48 horas antes da coleta da amostra sanguínea e que relatavam pelo menos um sintoma da doença. Foi realizada uma análise logística não condicional para avaliar a associação entre DFV e as características da população. Foram identificados um total de 122 (34,5%) casos da doença, entre os quais 39% eram fumantes e 61% eram do sexo masculino. As concentrações urinárias medianas de cotinina foram 75,6ng/mL (74,1ng/mg de creatinina) para não-casos e 755,8ng/mL (632,1ng/mg de creatinina) para os casos (valor de p ≤ 0,01). A análise multivariada mostrou uma associação positiva entre DFV e tempo de exposição à luz solar, exposição a pesticidas e pior estado de saúde geral, e uma associação inversa com cultivo de lenha. O estudo identificou alta prevalência de DFV e sugere o uso da cotinina como biomarcador significativo para identificar os casos de DFV, influenciando a distribuição por sexo. Uma vez que a fumicultura envolve danos à saúde, é importante implementar medidas para prevenir os impactos para os fumicultores, de acordo com os artigos 17 e 18 da Convenção-Quadro para o Controle do Tabaco da Organização Mundial da Saúde.

Resumen: Este estudio visó evaluar la ocurrencia de la enfermedad del tabaco verde (GTS por sus siglas en inglés) y sus factores asociados en pequeños agricultores de tabaco, que residen en Dom Feliciano, Rio Grande do Sul, Brasil. Se realizó un estudio transversal, evaluando la enfermedad del tabaco verde, en 354 pequeños agricultores de tabaco, entre octubre de 2011 y marzo 2012. La concentración urinaria de cotinina, un biomarcador de la exposición a la nicotina, se determinó durante el periodo de cosecha del tabaco. Los individuos que presentan niveles de cotinina urinarios ≥ 50ng/mL, que tuvieron contacto con hojas de tabaco hasta 48 horas antes de la recogida de la muestra, e informaron de al menos un síntoma de la enfermedad, fueron considerados casos. Se realizó un análisis logístico no condicional para evaluar la asociación entre GTS y las características de la población. Se identificaron un total de 122 (34,5%) casos, siendo un 39% de ellos fumadores y un 61% hombres. Las concentraciones medias de cotinina urinaria fueron 75,6ng/mL (74,1ng/mg de creatinina) para los no casos y 755,8ng/mL (632,1ng/mg de creatinina) para los casos (valor de p ≤ 0,01). El análisis multivariado mostró una asociación positiva entre el GTS y las horas de exposición a la luz solar, exposición a los pesticidas, un estatus peor de salud, y la asociación inversa con la cultura maderera. Este estudio presentó una alta prevalencia de GTS y sugiere que el uso de la cotinina urinaria es un biomarcador significativo para identificar casos de GTS, influenciando en la distribución por sexo. Debido a que la producción de tabaco implica perjuicios para la salud, es importante implementar medidas para prevenirlos en los agricultores del tabaco, como está estipulado en los artículos 17 y 18 del Convenio Marco para el Control del Tabaco de la Organización Mundial de la Salud.

Uso de biomarcador cotinina em trabalhadores para detecção da doença da folha verde do tabaco / Use of cotinine biomarker in workers to detect green tobacco sickness / Uso de biomarcador cotinina en trabajadores para detectar la Enfermedad del Tabaco Verde

Objetivo verificar, por meio da tecnologia de biomarcador de cotinina urinária, a ocorrência da doença da folha verde do tabaco em trabalhadores que cultivam tabaco do tipo Burley. Método estudo caso-controle pareado, com base no status tabágico e na proporção 1:4, com participação de 20 trabalhadores-caso e 91 controles. Para a coleta de dados foi realizada entrevista por meio de inquérito domiciliar e coleta de urina para exame de cotinina. Foram utilizados os testes t de Student, Mann-Whitney, qui-quadrado de Pearson ou exato de Fisher. Resultados dos 23 casos suspeitos, 20 apresentaram níveis elevados de cotinina, sinais e sintomas de dor de cabeça, irritação de pele, náusea, enjoo e mal-estar geral, principalmente pela manhã. A maioria trabalhou com tabaco molhado decorrente do sereno e o clima estava quente. Conclusão verificam-se sinais sugestivos da doença da folha verde do tabaco nos trabalhadores do tabaco Burley. Faz-se necessária a atuação de profissionais de saúde no desenvolvimento de ações promotoras de saúde e preventivas sobre essa doença relacionada ao trabalho.

Objective using the urinary cotinine biomarker to verify the occurrence of green tobacco sickness in workers who cultivate Burley tobacco. Method paired case-control study, based on smoking status and on the 1:4 ratio, with participation of 20 case workers and 91 controls. Data collection included household surveys and urine collection for cotinine examination. Student's T-Test, the Mann-Whitney test, Pearson's chi-square or Fisher's exact tests were used. Results of the 23 suspected cases, 20 showed elevated levels of cotinine, signs and symptoms of headache, skin irritation, nausea, sickness and general malaise, especially in the morning. Most had worked with tobacco that was wet from the morning dew and when the weather was warm. Conclusion there are signs suggestive of green tobacco sickness in Burley tobacco workers. The action of health professionals is necessary for the development of health promotion and preventive actions addressing work-related illness.

Objetivo certificar, por medio de la tecnología de biomarcador de cotinina urinaria, la ocurrencia de la Enfermedad del Tabaco Verde en trabajadores que cultivan tabaco del tipo Burley. Método estudio caso-control pareado, con base en el status tabáquico y en la proporción 1:4, con la participación de 20 trabajadores-caso y 91 controles. Para la recopilación de datos se realizó una encuesta por medio de cuestionario domiciliar y de recolección de orina para análisis de cotinina. Se utilizaron las siguientes pruebas t de Student, Mann-Whitney, chi-cuadrado de Pearson o exacta de Fisher. Resultados de los 23 casos sospechosos, 20 presentaron niveles elevados de cotinina, señales y síntomas de dolor de cabeza, irritación de la piel, náusea, mareo y malestar general, principalmente por la mañana. La mayoría trabajó con tabaco mojado por el sereno y en un clima caluroso. Conclusión se certifican señales sugestivas de Enfermedad del Tabaco Verde en los trabajadores del tabaco Burley. Se hace necesaria la actuación de profesionales de la salud en el desarrollo de acciones promotoras de salud y preventivas de esta enfermedad relacionado con el trabajo.

Self-reported smoking status and urinary cotinine levels in patients with asthma / Tabagismo entre asmáticos: avaliação por autorrelato e dosagem de cotinina urinária

ABSTRACT Objective: To determine the frequency of active smoking among patients with asthma and individuals without asthma by self-report and urinary cotinine measurement. Methods: This was a cross-sectional study conducted in the city of Salvador, Brazil, and involving 1,341 individuals: 498 patients with severe asthma, 417 patients with mild-to-moderate asthma, and 426 individuals without asthma. Smoking status was determined by self-report (with the use of standardized questionnaires) and urinary cotinine measurement. The study variables were compared with the chi-square test and the Kruskal-Wallis test. Results: Of the sample as a whole, 55 (4.1%) reported being current smokers. Of those, 5 had severe asthma, 17 had mild-to-moderate asthma, and 33 had no asthma diagnosis. Of the 55 smokers, 32 (58.2%) were daily smokers and 23 (41.8%) were occasional smokers. Urinary cotinine levels were found to be high in self-reported nonsmokers and former smokers, especially among severe asthma patients, a finding that suggests patient nondisclosure of smoking status. Among smokers, a longer smoking history was found in patients with severe asthma when compared with those with mild-to-moderate asthma. In addition, the proportion of former smokers was higher among patients with severe asthma than among those with mild-to-moderate asthma. Conclusions: Former smoking is associated with severe asthma. Current smoking is observed in patients with severe asthma, and patient nondisclosure of smoking status occurs in some cases. Patients with severe asthma should be thoroughly screened for smoking, and findings should be complemented by objective testing.

RESUMO Objetivo: Descrever a frequência de tabagismo ativo entre pacientes com asma e indivíduos sem asma, usando questionários padronizados e dosagem da cotinina urinária. Métodos: Estudo transversal realizado em Salvador (BA), com 1.341 indivíduos, sendo 498 com asma grave, 417 com asma leve/moderada e 426 sem asma. O tabagismo foi identificado por meio de autorrelato utilizando questionários e por mensuração da cotinina urinária. Para a comparação das variáveis estudadas, utilizaram-se os testes do qui-quadrado e de Kruskal-Wallis. Resultados: Dos 55 participantes (4,1%) que se declararam tabagistas atuais, 5, 17 e 33 eram dos grupos asma grave, asma leve/moderada e sem asma, respectivamente. Desses 55, 32 (58,2%) eram tabagistas diários e 23 (41,8%) eram tabagistas ocasionais. Observaram-se níveis elevados de cotinina urinária entre não fumantes autodeclarados e tabagistas pregressos, especialmente no grupo asma grave, o que sugere omissão do hábito atual de fumar. A carga tabágica entre os fumantes e a proporção de ex-tabagistas foram maiores no grupo asma grave do que no grupo asma leve/moderada. Conclusões: O tabagismo pregresso esteve associado à asma grave. Tabagismo atual também foi observado em alguns pacientes com asma grave e detectou-se omissão em alguns casos. A investigação de tabagismo deve ser meticulosa em pacientes com asma grave e a entrevista desses deve ser complementada por uma avaliação objetiva.

Intranasal Cotinine Plus Krill Oil Facilitates Fear Extinction, Decreases Depressive-Like Behavior, and Increases Hippocampal Calcineurin A Levels in Mice.

Failure in fear extinction is one of the more troublesome characteristics of posttraumatic stress disorder (PTSD). Cotinine facilitates fear memory extinction and reduces depressive-like behavior when administered 24 h after fear conditioning in mice. In this study, it was investigated the behavioral and molecular effects of cotinine, and other antidepressant preparations infused intranasally. Intranasal (IN) cotinine, IN krill oil, IN cotinine plus krill oil, and oral sertraline were evaluated on depressive-like behavior and fear retention and extinction after fear conditioning in C57BL/6 mice. Since calcineurin A has been involved in facilitating fear extinction in rodents, we also investigated changes of calcineurin in the hippocampus, a region key on contextual fear extinction. Short-term treatment with cotinine formulations was superior to krill oil and oral sertraline in reducing depressive-like behavior and fear consolidation and enhancing contextual fear memory extinction in mice. IN krill oil slowed the extinction of fear. IN cotinine preparations increased the levels of calcineurin A in the hippocampus of conditioned mice. In the light of the results, the future investigation of the use of IN cotinine preparations for the extinction of contextual fear memory and treatment of treatment-resistant depression (TRD) in PTSD is discussed.

Cotinine: A Therapy for Memory Extinction in Post-traumatic Stress Disorder.

Post-traumatic stress disorder (PTSD) is a mental disorder that may develop after exposure to exceptionally threatening or unescapable horrifying events. Actual therapies fail to alleviate the emotional suffering and cognitive impairment associated with this disorder, mostly because they are ineffective in treating the failure to extinguish trauma memories in a great percentage of those affected. In this review, current behavioral, cellular, and molecular evidence supporting the use of cotinine for treating PTSD are reviewed. The role of the positive modulation by cotinine of the nicotinic acetylcholine receptors (nAChRs) and their downstream effectors, the protection of astroglia, and the inhibition of microglia in the PTSD brain are also discussed.

Cotinine Plus Krill Oil Decreased Depressive Behavior, and Increased Astrocytes Survival in the Hippocampus of Mice Subjected to Restraint Stress.

Restraint stress (RS) is a condition affecting millions of people worldwide. The investigation of new therapies to alleviate the consequences of prolonged RS is much needed. Cotinine, a nicotine-derivative, has shown to prevent the decrease in cerebral synaptic density, working memory deficits, anxiety, and depressive-like behavior after prolonged restraint stress (RS) in mice. Furthermore, post-treatment with cotinine reduced the adverse effects of chronic RS on astrocyte survival and architecture. On the other hand, the nutritional supplement krill oil (KO), has shown to be beneficial in decreasing depressive-like behavior and oxidative stress. In this study, in the search for effective preventative treatments to be used in people subjected to reduced mobility, the effect of co-treatment with cotinine plus KO in mice subjected to prolonged RS was investigated. The results show that cotinine plus KO prevented the loss of astrocytes, the appearance of depressive-like behavior and cognitive impairment induced by RS. The use of the combination of cotinine plus KO was more effective than cotinine alone in preventing the depressive-like behavior in the restrained mice. The potential use of this combination to alleviate the psychological effects of reduced mobility is discussed.

Urinary cotinine in tobacco farmers in Southern Brazil

ABSTRACT OBJECTIVE To describe urinary cotinine levels in tobacco farmers. METHODS A cross-sectional study was conducted in 2,570 tobacco farmers. All participants that reported green tobacco sickness in the week prior to the interview plus a subsample of 492 pesticide applicators were included. We collected urinary samples and information about sociodemographic, behavioral, dietary, occupational characteristics, and pesticide poisoning during their lifetime. Stratification by sex and smoking was performed and the Wilcoxon and Kruskal-Wallis non-parametrical tests were used to analyze cotinine means. RESULTS This study included 582 individuals. There was no difference in urinary cotinine means between green tobacco sickness symptomatic and asymptomatic individuals. Among non-smokers, having picked tobacco in the previous week was associated with higher cotinine means in both genders. Cotinine levels were higher on the first day of symptoms and reduced exponentially with each day in female non-smokers. Male non-smokers had higher levels on the second day and a more gradual reduction. The cotinine level rose up to 15 cigarettes/day of consumption. CONCLUSIONS The urinary cotinine measures exposure to nicotine up to its saturation point; while green tobacco sickness, affected by tolerance, indicates nicotine poisoning. Strategies to reduce nicotine exposure in tobacco production are needed. Mechanization could be an alternative, as long as it overcame the challenge of irregular terrain and did not affect the quality of the leaf. More studies are needed to evaluate the chronic effect of nicotine exposure.

Secondhand tobacco smoke exposure and pulmonary function: a cross-sectional study among non-smoking employees of bar and restaurants in Santiago, Chile.

INTRODUCTION: The workplace remains a significant source of secondhand smoke (SHS) exposure. This pollutant is known to be associated with respiratory and cardiovascular problems, but its effects on specific pulmonary function parameters remain largely unexplored. The objectives of this study were to measure SHS exposure among non-smoking employees of bar and restaurants in Santiago, Chile and to evaluate the effects of such exposure on pulmonary function. METHODS: Cross-sectional design. The study sample included non-smoking workers from 57 restaurants and bars in Santiago, Chile. The outcome variable was pulmonary function and the exposure variables were urine cotinine concentration, a biomarker for current SHS exposure, and years of SHS exposure in the workplace as proxy of chronic exposure. Personal and occupational variables were also recorded. Data analysis was performed using linear regression models adjusted by confounders. RESULTS: The median age of the workers was 35 years and the median employment duration at the analysed venues was 1 year. Workers in smoking facilities reported greater SHS exposure (36 hours per week) than workers in smoke-free locations (4 hours per week). Urine cotinine levels were inversely correlated with forced vital capacity, but the finding was not statistically significant (ß=-0.0002; 95% CI -0.007 to 0.006). Years of exposure to SHS showed to be significantly associated with forced expiratory flow25/75 (ß=-0.006; 95% CI -0.010 to -0.0004). CONCLUSION: These findings suggest that cumulative exposure to SHS at work may contribute to deterioration of pulmonary function in non-smoking employees.

Low cigarette smoking prevalence in peri-urban Peru: results from a population-based study of tobacco use by self-report and urine cotinine.

BACKGROUND: A recent study found lower self-reported prevalence of tobacco smoking in a peri-urban area of Lima, Peru than previously reported in urban samples. These regions encompass substantial proportions of Peru's population - ones at greater risk of disease due to reduced healthcare access - but have been less often studied. We validate low smoking prevalence with urine cotinine and characterize chronic disease and lung function outcomes between non-, occasional, and daily smokers. METHODS: Data are from the CRONICAS Cohort Study, a population-based longitudinal study in four low-resource Peruvian settings, which began in 2010. Of a baseline cohort of 2978 adults, we prospectively followed 2583 (87%) to determine prevalence of chronic illness. RESULTS: In a baseline sub-sample of 382 participants, median adjusted cotinine was 0.0 mcg/mg (IQR 0-0) for both self-reported non-smokers and occasional smokers compared to 172.3 mcg/mg (IQR 0-709.2) for daily smokers. Creatinine-adjusted cotinine validated daily smoking prevalence of 4.7% at a cutoff of 100 mcg/mg. Kappa statistic for daily smoking and creatinine- adjusted cotinine ≥100 mcg/mg was 0.65 (95% CI 0.47, 0.83), indicating substantial agreement. At baseline, we found 3.3% daily and 8.9% occasional smoking by self-report for the full cohort. Follow-up indicated little difference in chronic disease prevalence between groups. Daily smokers trended toward having a greater decline in FVC (-1%; 95% CI -2.9, 0.8) and FEV1 (-1.3%; 95% CI -3.2, 0.6) over 40 months when compared to non-smokers, whereas the decline in lung function for occasional smokers was similar compared to non-smokers (-0.2% FVC; 95% CI -1.5, 1.0) and (0% FEV1; 95% CI -1.3, 1.3). CONCLUSIONS: Our data places Peru within a previously-described pattern of smoking found in much of Latin America, favoring occasional over daily smoking and low cigarette consumption. We determine that there are not significant differences between smoking groups concerning chronic disease outcomes. We favor distinguishing between daily and occasional smokers in order to accurately characterize these low-use populations.