Altered DNA methylation may lead to suboptimal fetal programming, increasing the risk of adverse pregnancy outcomes such as small for gestational age (SGA); however, few studies have examined the associations between DNA methylation, prenatal exposures, and fetal outcomes. Cross-sectional data from a larger, ongoing study were used to assess the impact of prenatal smoking on gene specific methylation of umbilical cord blood derived DNA and to investigate the association between gene-specific methylation and risk of SGA. The association between gene-specific DNA methylation and birthweight was also assessed. Maternal and infant covariates were abstracted from medical records, cigarette smoke exposure was determined by measuring cotinine in umbilical cord blood plasma, and the Illumina Infinium Methylation27 assay was used to assess CpG site specific methylation. Methylation was represented by a beta value ranging from 0 to 1. Gene-level methylation was calculated by averaging the methylation levels over the CpG sites interrogated in that gene. Logistic regression was used to generate adjusted odds ratios (OR) and 95% confidence intervals (CI) for the association between SGA and methylation of CYP1A1, HIF1A, GSTT1, and GSTM1 and the association between cotinine level and hypermethylation of CYP1A1, HIF1A, GSTT1, and GSTM1. DNA was considered hypermethylated if the beta value was greater than or equal to the 75th percentile. Univariate and multivariable linear regression were used to examine the association between birthweight and methylation of the IGF1 and IGF2 gene. The analyses included 90 singleton births. A 0.10 unit increase in methylation of GSTT1 increased the risk of SGA almost 3-fold (OR=2.69, 95%CI=1.34, 5.43). A 5ng/ml increase in cotinine level increased the risk of hypermethylation of GSTT1 (OR=1.18, 95%CI=1.02, 1.37). Birthweight did not appear to be impacted by methylation of IGF2 (β=0.07, 95%CI=-2.91, 3.05), but a one standard deviation increase in methylation of IGF1 was associated with a 3.63% decrease in birthweight (95%CI= -6.49, -0.78). No differences in DNA methylation by prenatal vitamin intake were detected. These findings suggest that DNA methylation plays a critical role in fetal growth and may mediate the risk of SGA and low birthweight.

[Truncated abstract] With the dramatic rise in asthma and allergic disease there is an urgent need to define the early life exposures which influence developing immune responses to increase the predisposition to allergic disease. While this is clearly multifactorial, this thesis addresses the effects of maternal smoking as a major adverse, yet avoidable exposure in early life. I hypothesised that the well-documented increased susceptibility to infection in infants of smokers could indicate underlying effects on innate Toll-like receptor (TLR) mediated microbial responses which could in turn contribute to early immune dysregulation and increased risk of allergic disease. In addition to providing the first defence against microbes, TLR-mediated pathways modulate subsequent specific immune response and are of growing interest in the potential inhibition of inappropriate allergic responses. My initial interest in the potential immune effects of smoking in pregnancy was based on preliminary retrospective analyses of a previous cohort (presented in Chapter 3) which suggested possible effects on T cell cytokine responses to mitogens and allergens. Based on this, I recruited a new prospective pregnancy cohort (n=122) of smokers (n=60) and non-smokers (n=62) (as outlined in Chapter 4) to confirm this and test my novel hypothesis that maternal smoking may be affecting important innate (TLR-mediated) immune pathways. … Thus, these findings could indicate that smoking increases the early susceptibility to infection thereby increasing subsequent IgA responses. This is supported by observations that key neonatal TLR responses are attenuated in children who go onto develop wheezy illnesses and lower respiratory tract infections. Together, the study findings suggest that in addition to effects on lung growth, maternal smoking may also influence aspects of neonatal innate immune function that are now believed to play a critical role in microbial-driven postnatal immune development, highlighting that other environmental interactions are also highly relevant to the v

Objetivos: Diversos trabalhos documentaram a associação entre o fumo ativo na gestação e a redução no peso,comprimento e perímetro cefálico dos recémnascidos. Porém, existem controvérsias quanto aos efeitos do fumo passivo. Esse estudo buscou colaborar na elucidação sobre os efeitos da exposição da gestante ao fumo nas medidas antropométricas do recém-nascido. Métodos: Estudo transversal, que incluiu 200 parturientes, identificando a intensidade da exposição das mesmas ao fumo através da mensuração da cotinina urinária e do auto-relato sobre seu hábito tabágico e de seus familiares,o que possibilitou categorizá-las em não fumantes, fumantes passivas e fumantes ativas e posterior comparação com as medidas antropométricas dos recémnascidos. Resultados: Encontramos uma diferença estatisticamente significativa no peso,quando comparamos as fumantes passivas às não fumantes, -152g (IC95%: -285 a -18;P=0,026) e a diferença encontrada nas fumantes ativas foi de - 159g (IC95%: -301 a -16; P=0,029), em relação às não fumantes. Na medida do comprimento, observou-se uma diferença significativa entre as fumantes passivas, -0,8cm (IC95%: -1,4 a -0,2;P=0,012) em relação às não fumantes, e entre as fumantes ativas a diferença foi de -1,0cm (IC95%: -1,6 a -0,3;P=0,003), quando comparado às não fumantes. Quanto à medida do perímetro cefálico, não foram observadas diferenças significativas quando os grupos, das fumantes passivas e fumantes ativas, foram comparados ao grupo das não fumantes. Conclusões: Os resultados sugerem que o fumo passivo durante a gestação pode apresentar efeitos semelhantes aos do fumo ativo nas medidas antropométricas dos recém-nascidos. / Objectives: Several studies have documented the association between active smoking during pregnancy to weight, height and head circumference reduction on newborns. However, there are controversies about the passive smoking. The present study sough the elucidation of the effects of tobacco exposition of pregnant women on anthropometric measurements of newborns. Methods: Cross-sectional study, including 200 in labor women, identifying the intensity of their exposition to tobacco through the measurement of urine cotinine and the self reported information about the smoking habit of their relatives and themselves, what allowed us to classify them as non-smokers, passive smokers and active smokers and later the comparison with the anthropometric measurements of newborns. Results: We found a statistically significant difference in weight when comparing passive smokers to non-smokers, -152g (CI 95%: -285 to –18; P=0.026) and the difference found on active smokers was - 159g (CI 95%: -301 to 16; P=0.029) when compared to non-smokers. On the height measurements, a significant difference was found within the group of passive smokers, -0,8 cm (CI 95%: -1,4 to -0.2; P=0.012) comparing to the non-smokers group, within the smokers group the difference was -1,0 cm (CI 95%: -1,6 to -0.3; P=0.003) when compared to nonsmokers. About the measurements of head circumference, no was found a significant difference when the passive smokers and active smokers group was compared to non-smokers group. Conclusions: The results suggest that the passive smoking during pregnancy may present similar effects as the active smoking on the anthropometric measurements of newborns.

Objetivos: Diversos trabalhos documentaram a associação entre o fumo ativo na gestação e a redução no peso,comprimento e perímetro cefálico dos recémnascidos. Porém, existem controvérsias quanto aos efeitos do fumo passivo. Esse estudo buscou colaborar na elucidação sobre os efeitos da exposição da gestante ao fumo nas medidas antropométricas do recém-nascido. Métodos: Estudo transversal, que incluiu 200 parturientes, identificando a intensidade da exposição das mesmas ao fumo através da mensuração da cotinina urinária e do auto-relato sobre seu hábito tabágico e de seus familiares,o que possibilitou categorizá-las em não fumantes, fumantes passivas e fumantes ativas e posterior comparação com as medidas antropométricas dos recémnascidos. Resultados: Encontramos uma diferença estatisticamente significativa no peso,quando comparamos as fumantes passivas às não fumantes, -152g (IC95%: -285 a -18;P=0,026) e a diferença encontrada nas fumantes ativas foi de - 159g (IC95%: -301 a -16; P=0,029), em relação às não fumantes. Na medida do comprimento, observou-se uma diferença significativa entre as fumantes passivas, -0,8cm (IC95%: -1,4 a -0,2;P=0,012) em relação às não fumantes, e entre as fumantes ativas a diferença foi de -1,0cm (IC95%: -1,6 a -0,3;P=0,003), quando comparado às não fumantes. Quanto à medida do perímetro cefálico, não foram observadas diferenças significativas quando os grupos, das fumantes passivas e fumantes ativas, foram comparados ao grupo das não fumantes. Conclusões: Os resultados sugerem que o fumo passivo durante a gestação pode apresentar efeitos semelhantes aos do fumo ativo nas medidas antropométricas dos recém-nascidos. / Objectives: Several studies have documented the association between active smoking during pregnancy to weight, height and head circumference reduction on newborns. However, there are controversies about the passive smoking. The present study sough the elucidation of the effects of tobacco exposition of pregnant women on anthropometric measurements of newborns. Methods: Cross-sectional study, including 200 in labor women, identifying the intensity of their exposition to tobacco through the measurement of urine cotinine and the self reported information about the smoking habit of their relatives and themselves, what allowed us to classify them as non-smokers, passive smokers and active smokers and later the comparison with the anthropometric measurements of newborns. Results: We found a statistically significant difference in weight when comparing passive smokers to non-smokers, -152g (CI 95%: -285 to –18; P=0.026) and the difference found on active smokers was - 159g (CI 95%: -301 to 16; P=0.029) when compared to non-smokers. On the height measurements, a significant difference was found within the group of passive smokers, -0,8 cm (CI 95%: -1,4 to -0.2; P=0.012) comparing to the non-smokers group, within the smokers group the difference was -1,0 cm (CI 95%: -1,6 to -0.3; P=0.003) when compared to nonsmokers. About the measurements of head circumference, no was found a significant difference when the passive smokers and active smokers group was compared to non-smokers group. Conclusions: The results suggest that the passive smoking during pregnancy may present similar effects as the active smoking on the anthropometric measurements of newborns.

Data de aprova??o ausente. / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2016-12-20T14:07:54Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) morgana_fernandes_alecrim.pdf: 1686041 bytes, checksum: c2250fb532be684ddef616d3424c8dc0 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-01-17T18:51:10Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) morgana_fernandes_alecrim.pdf: 1686041 bytes, checksum: c2250fb532be684ddef616d3424c8dc0 (MD5) / Made available in DSpace on 2017-01-17T18:51:10Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) morgana_fernandes_alecrim.pdf: 1686041 bytes, checksum: c2250fb532be684ddef616d3424c8dc0 (MD5) Previous issue date: 2016 / A Cotinina (CO) ? o principal metab?lito da nicotina, sendo utilizada como biomarcador para monitorar a exposi??o de um indiv?duo ao tabaco. Os m?todos comumente utilizados para a determina??o desse composto em amostras biol?gicas s?o baseados na cromatografia l?quida, os quais, em geral, s?o dispendiosos e requerem tratamentos tediosos dessas amostras. Dessa maneira, torna-se necess?rio o desenvolvimento de novos m?todos mais simples e r?pidos para determina??o da CO. Nesse contexto, o presente trabalho prop?e o uso eletrodo de diamante dopado com boro (BDD) para detec??o e caracteriza??o eletroqu?mica da CO por t?cnicas voltam?tricas, bem como sua quantifica??o em amostras de saliva por meio da amperometria de m?ltiplos pulsos (MPA) em sistema de an?lise por inje??o em fluxo (FIA). Em meio de tamp?o fosfato a 0,1 mol L-1 (pH 7) a CO apresentou dois processos eletroqu?micos, um an?dico em +1,8 V e um cat?dico em -1,2 V (vs. Ag/ AgCl), sendo o segundo dependente do primeiro e ambos de car?ter irrevers?vel. Foi verificado que o processo de transporte de massas para a oxida??o da CO ? preferencialmente difusional sobre o eletrodo de BDD e que 2 el?trons e pr?tons est?o envolvidos na rea??o eletroqu?mica. O coeficiente de difus?o calculado para a CO foi de 5,7 x 10 -2 cm2 s-1. Para determina??o da CO em saliva, MPA foi utilizada aplicando-se 4 pulsos de potencial, sendo: (1) em +1,8 V/ 400 ms potencial gerador (EG) para oxida??o da CO; (2) em -1,0 V/ 500 ms potencial para elimina??o de interferentes na amostra de saliva, (3) em -1,2 V/ 30 ms potencial coletor para reduzir e quantificar a CO gerada pelo EG; (4) em -145/ 300 ms potencial para limpeza da superf?cie do BDD. A freq??ncia anal?tica te?rica foi obtida de 24 determina??es por hora com uma vaz?o otimizada em 3,0 mL min-1 e a al?a de amostragem de 300 ?L no sistema FIA. Al?m disso, um baixo desvio padr?o relativo de 1,46% foi obtido para 10 determina??es consecutivas de CO 10 ?mol L-1. Nessas condi??es, foi obtida uma faixa linear de 0,5 a 100 ?mol L-1 (R= 0,998) com limite de detec??o calculado em 0,18 ?mol L-1 para CO. Os estudos de adi??o e recupera??o da CO em amostras de saliva foram de 96,77%, Portanto, mediante a caracteriza??o eletroqu?mica e os estudos por FIA-MPA usando eletrodo de BDD, este trabalho apresenta uma alternativa simples, r?pida e de baixo custo para monitoramento do grau de exposi??o ao tabaco pela determina??o da CO em amostras de saliva. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, [2016]. / The Cotinine (CO) is the major metabolite of the nicotine an disused as a biomarker to monitor the exposure of an individual to tobacco. The methods commonly used for CO determination in biological samples are based on liquid chromatography, which generally are expensive and require tedious treatments of the samples. Thus, it becomes necessary to develop simpler and faster methods for determining CO. In this context, this paper proposes the use boron-doped diamond (BDD) film electrode for the detection and electrochemical characterization of CO by voltammetric techniques and, CO quantification in saliva samples by multiple-pulse amperometry (MPA) in analysis by flow injection (FIA) system. In phosphate buffer solution 0.1 mol L-1 (pH 7), the CO presented two electrochemical processes, one anodic at + 1.8V and one cathode at -1.2V (vs. Ag/ AgCl), with the second process dependent on the first and both irreversible process. It was found that CO electrochemical processes are controlled by diffusion and, two-protons and two-electrons are involved in the oxidation reaction. The diffusion coefficients calculated for CO was 5.7 x 10 -2 cm2 s-1. In determination of CO in saliva samples, the MPA detection was used applying four potential pulses: (1) at +1.8V/ 400ms ? generator potential pulse (EG) for oxidation of CO; (2) at-1.0V/500ms - potential for remove possible interferents, (3) at - 1.2V/30ms - collector potential pulse potential for reduction of the product generated by EG and CO quantification; (4) at-14.5V/ 300 ms- potential pulse for cleaning BDD electrode. The proposed method obtained a theoretical analytical frequency with 24 determinations per hour by FIA system in flow rate of 3.0 ml min-1 and sample loop of 300?L. Furthermore, a low relative standard deviation of 1.46% was obtained for 10 measurements of CO 10 ?mol L-1. Under these conditions, A good linear range from 0.5 to 100 ?mol L-1 (R = 0.998) was obtained with a detection limit estimated at 0.18 ?mol L-1 for CO. The addition and recovery studies of CO in saliva samples were 96.77%. Therefore, this work presents, by the first time, an electrochemical characterization of the CO and a method simple, fast and low cost for monitoring the degree of exposure to tobacco by CO determination in saliva samples using FIA-MPA technique.

Indiana University-Purdue University Indianapolis (IUPUI) / Analyses of tobacco products and their usage are well-researched and have implications in analytical chemistry, forensic science, toxicology, and medicine. As such, analytical methods must be developed to extract compounds of interest from tobacco products and biological specimens in order to determine tobacco exposure. In 2009, R.J. Reynolds Tobacco Co. released a line of dissolvable tobacco products that are marketed as a smoking alternative. The dissolvables were extracted and prepared by ultrasonic extractions, derivatization, and headspace solid phase microextraction (SPME) with analysis by gas chromatography-mass spectrometry (GC-MS). The results show that the compounds present are nicotine, flavoring compounds, humectants and binders. Humectant concentrations vary among different tobacco types depending on the intended use. Humectants were quantified in various tobacco types by GC and “splitting” the column flow between a flame ionization detector (FID) and an MS using a microfluidic splitter in order to gain advantage from the MS’s selectivity. The results demonstrated excellent correlation between FID and MS and show that MS provides a higher level of selectivity and ensures peak purity. Chemometrics was also used to distinguish products by tobacco type. Hair is a common type of evidence in forensic investigations, and it is often subjected to mitochondrial DNA (mtDNA) analysis. Preliminary data was gathered on potential “lifestyle” markers for smoking status as well as any indications of subject age, gender, or race by investigating the organic “waste” produced during a mtDNA extraction procedure. The normally discarded organic fractions were analyzed by GC-MS and various lipids and fatty acids were detected. At this point, a total vaporization-SPME (TV-SPME) method was theorized, developed, and optimized for the specific determination of nicotine and its metabolite, cotinine. The theory of TV-SPME is to completely vaporize an organic extract which will eliminate the partitioning between the sample and the headspace, thereby simplifying the thermodynamic equilibrium. Parameters such as sample volume, incubation temperature, and extraction time were optimized to achieve the maximum analyte signal. Response surface methodology (RSM) is a statistical model that is very useful in predicting and determining optimum values for variables to ensure the ideal response. RSM was used to optimize the technique of TV-SPME for the analysis of nicotine and cotinine. Lastly, quantitation of nicotine and cotinine in human hair typically requires large sample sizes and extensive extraction procedures. Hence, a method using small sample sizes and a simple alkaline digestion followed by TV-SPME-GC-MS has been developed. Hair samples were collected from anonymous volunteers and nicotine and cotinine were identified and quantitated in the hair of tobacco users.

The relevance of preconceptional and prenatal toxicant exposures for genomic stability in offspring is difficult to analyze in human populations, because gestational exposures usually cannot be separated from preconceptional exposures. To analyze the roles of exposures during gestation and conception on genomic stability in the offspring, stability was assessed via the Comet assay and highly sensitive, semiautomated confocal laser scans of gammaH2AX foci in cord, maternal, and paternal blood as well as spermatozoa from 39 families in Crete, Greece, and the United Kingdom. With use of multivariate linear regression analysis with backward selection, preconceptional paternal smoking (% tail DNA: P>0.032; gammaH2AX foci: P>0.018) and gestational maternal (% tail DNA: P>0.033) smoking were found to statistically significantly predict DNA damage in the cord blood of F1 offspring. Maternal passive smoke exposure was not identified as a predictor of DNA damage in cord blood, indicating that the effect of paternal smoking may be transmitted via the spermatozoal genome. Taken together, these studies reveal a role for cigarette smoke in the induction of DNA alterations in human F1 offspring via exposures of the fetus in utero or the paternal germline. Moreover, the identification of transgenerational DNA alterations in the unexposed F1 offspring of smoking-exposed fathers supports the claim that cigarette smoke is a human germ cell mutagen.

No description available.

Le tabagisme est reconnu comme une dépendance comparable aux autres dépendances : alcool, opiacées et autres psycho-stimulants. Les mécanismes responsables de l'initiation et du maintien de l'addiction sont également impliqués dans les déviations comportementales en général, comme les déviations nutritionnelles, les compulsions.... La nicotine de la feuille de tabac est très toxique, dès son absorption, elle atteint le cerveau et tout l'organisme, active ses récepteurs et produit des effets toxiques et des adaptations homéostatiques. L'importance de ses effets va dépendre de la dose, du mode d'administration, de la chronicité, de l'effet considéré, du génome considéré et des interactions avec son principal dérivé, la cotinine. La cotinine résulte de l'addition d'un atome d'oxygène en position α du noyau pyrrolidine. Les conséquences de cette métabolisation ont été évaluées, dans l'étude présente, en partant des structures chimiques de ces deux alcaloïdes jusqu'à l'isolement des mécanismes biochimiques, neurochimiques, moléculaires et comportementaux de leurs actions. Ces différents mécanismes ont été validés par une étude intégrative en pointant le monoxyde d'azote NO comme un médiateur de la dépendance tabagique, en accord avec les données de l'étude bibliographique qui impliquent ce même médiateur dans toutes les dépendances. Les mécanismes à la base des prédispositions, le début et les raisons de l'évolution vers un état dépendant et les raisons des rechutes font l'objet d'intenses investigations. Les travaux dans ce domaine suggèrent que certaines modifications des protéines transmettent un signal de longue durée et que les espèces réactives de l'oxygène et du nitrogène sont à la base des mécanismes de potentialisation et de dépression à long terme. Notre travail montre une absence de toxicité pour la cotinine [422,423], une activité psychostimulante pure [414], une pharmacologie nouvelle non nicotinique [416,417,419,422,423], un passage actif [415] dans le cerveau régulé par le système nicotinique périphérique [412,421], la forme endogène et exogène de la cotinine [424], la médiation d'activité anti stress du récepteur p40 de la cotinine [420] et son homologie avec les protéines humaines impliquées dans les réactions inflammatoires [36,161], stimulatrice paracrine de la croissance cellulaire [55], un rôle dans la libération de dopamine, la production d'un stress oxydant par l'administration de la cotinine [418], un renforcement dans le contexte des approches flexibles, la participation forte des états émotionnels et d'anxiété à l'action anxiolytique de l'administration et anxiogène du retrait de la cotinine. Ils permettent de proposer que la nicotine agit directement et indirectement par sa conversion en cotinine. Cette dernière agit par ses récepteurs, au niveau central par la p40, pour moduler les taux de dopamine avec des conséquences sur l'apprentissage, la récompense, le stress oxydatif, l'anxiété et les réponses potentialisées et déprimées à long terme. Comme conclusion, nos travaux permettent de proposer de nouvelles cibles pharmacologiques, méthodes et concepts permettant de comprendre à l'échelle biochimique, neurochimique, moléculaire et comportemental l'addiction tabagique. L'espoir est d'utiliser ces connaissances pour différencier les susceptibilités et développer de nouvelles approches préventives et thérapeutiques.