O mecônio é uma matéria fecal que começa a acumular no intestino do feto por volta do terceiro mês de gestação e normalmente é eliminado nos primeiros dias de vida do recém nascido. No mecônio ocorre o acúmulo de substâncias com as quais a mãe entrou em contato durante o período de gestação e, portanto, sendo possível avaliar a exposição fetal. Nos casos de mães fumantes, compostos presentes no tabaco tais como nicotina e substâncias que são formadas após a metabolização da nicotina como por exemplo a cotinina, podem ser também detectadas nas amostras de mecônio, já que ocorre o acúmulo de nicotina e seus metabólitos no mesmo. O uso do cigarro durante o período gestacional acarreta em uma série de problemas ao feto como baixo peso ao nascer, parto pré-maturo, doenças no trato respiratório, dentre outros. Nos dias atuais é possível fazer a pesquisa de drogas lícitas e ilícitas em várias amostras biológicas tais como, sangue, urina, cabelo, fluido oral, mecônio, entre outras. O mecônio é uma boa opção, por vários motivos: amostragem fácil e não invasiva (a coleta pode ser feita na fralda), indica o histórico do uso de substâncias pela mãe durante a gestação por ser cumulativo, etc. O objetivo foi desenvolver e validar um método analítico empregando cromatografia em fase gasosa acoplada a espectrometria de massas para a determinação de nicotina e, seu metabólito, cotinina em amostras de mecônio coletadas de recém nascidos. Para o desenvolvimento do método foi utilizado 0,5g de mecônio e os analitos foram extraídos com metanol e em seguida a amostra foi submetida a purificação através da extração em fase sólida utilizando cartuchos Bond Elut Certify I. O método foi validado de acordo com os parâmetros estabelecidos pela ANVISA e demonstrou linearidade de 160 1600 ng/g para nicotina e de 160 1000 ng/g para cotinina. Os limites de detecção estabelecidos foram de 10 ng/g para nicotina e 60 ng/g para cotinina, enquanto os limites de quantificação foram de 60 ng/g para nicotina e 100 ng/g para cotinina. A exatidão apresentou valores de 91,73% a 103,73%, a precisão intra-ensaio variou entre 3,21% a 10,86%, e a precisão inter-ensaio obteve valores entre 4,91% a 9,88%. O método validado foi aplicado em amostras de mecônio coletadas no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP-USP). / Meconium is a fecal matter passed by the newborn after birth. It begins to form around the 3th month of gestation and accumulates in the fetus until birth. Substances which the mothers had contact during the gestation period may accumulate in meconium and, therefore, its possible to assess fetal exposure. Substances from the tobacco smoke, like nicotine and its metabolite cotinine, also accumulates in the meconium and can be detected .Maternal smoking during pregnancy is hazardous to the fetus and it is associated with low birth weight, prematurity, respiratory tract infections and others. Nowadays it is possible to assess licit and illicit drugs in several biological samples like blood, urine, hair, oral fluid, meconium and others. Meconium is the best choice because its easy to collect and its noninvasive, indicates a history of substances use by the mother in the latter half of pregnancy, etc. The purpose of this study was to develop and validate a methodology using gas chromatography-mass spectrometry to assess nicotine and cotinine in meconium samples collected from newborns. To the development of the methodology 0,5g of meconium was used per assay and the analytes were extracted from the matrix using methanol. Then, a solid phase extraction was applied using Bond Elut Certify I cartridges. The methodology was validated in the range of 160 1600 ng/g for nicotine and 160 - 1000 ng/g for cotinine. The limit of detection established was 10 ng/g for nicotine and 60 ng/g for cotinine, while the limit of quantification was 60 ng/g for nicotine and 100 ng/g for cotinine. The accuracy showed values between 91,73% and 103,73%, the intra-assay precision between 3,21% and 10,86% and the inter-assay precision between 4,91% and 9,88%. The validated methodology was applied to analysis of meconium samples collected from newborns in the Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP-USP).

Pediatric obesity has reached epidemic proportions. Reducing obesity among children is expected to lower their likelihood of being obese as adults and, therefore, lower their cardiovascular and metabolic risk profile in adulthood including hypertension, dyslipidemia, type II diabetes, heart disease, and stroke. Pediatric obesity among ages 2–19 is defined as a body mass index (BMI) greater than or equal to the 95th percentile for age and gender as defined according to the CDC BMI-for-age growth charts. Risk factors for obesity are present as early as birth, suggesting exposures at different stages of the life cycle are important to study. The primary objective of this thesis was to evaluate exposures throughout childhood and evaluate their association with both weight and metabolic status. Study 1 examined the relationship between physical activity and metabolic syndrome in overweight and obese youth ages 12–19. We found that even modest amounts of moderate to vigorous physical activity were associated with a reduction in risk of metabolic syndrome, with time spent in vigorous physical activity driving the association. Study 2 explored the relationship between environmental tobacco smoke (ETS) exposure and childhood overweight and obesity in 3–6 year old children. We observed that ETS has a positive association with risk of overweight/obesity, with a dose-response effect observed. Study 3 examined the relationship between maternal antibiotic use during pregnancy and infant birthweight. We did not observe any association between maternal antibiotic use and birthweight or BW/GA-z (birthweight adjusted for gestational age z-score), but we did observe a reduction in risk of SGA (small for gestational age) for infants exposed to antibiotics during gestation. This association was most evident among third trimester users. / 2019-06-09T00:00:00Z

This dissertation mainly focuses on applications of Surface Enhance Raman Scattering (SERS) to detect tobacco-related biomarkers with optimized experimental conditions (pH and aggregating agents) and SERS substrates (silica core and silver shell nanoparticles). Cotinine (COT) and trans 3-hydroxycotinine (3HC), metabolized from nicotine as one of main chemicals of tobacco, have been used as tobacco biomarkers because their half-life are longer than that of nicotine, which enable to monitor the tobacco exposure. The effects of aggregating agents and pH on SERS detection of COT and 3HC were investigated. Aggregating agents play an important role in SERS detection of target molecules since the strong SERS enhancements are observed from junctions of nanoparticles which can be induced by aggregating agents, and so called "hot spot". That is, the more hot spots are created among the nanoparticles by aggregating agents, the higher the SERS enhancement is. Five cationic (K+, Na+, Mg2+, Li+, Ca2+) and three anionic (Cl-, Br-, I-) aggregating agents were tested. Interestingly but not surprisingly, optimal concentrations of 11 kinds of aggregating agents for COT and 3HC detections vary dramatically within two orders of magnitude. In addition, the effect of pH conditions on SERS intensity of COT and 3HC was investigated since the protonated or deprotonated molecules induced by various ranges of pH values produces change in SERS intensity of the molecule. The highest SERS enhancement is obtained using 1.5 mM MgCl2 for COT at pH 7 and 50 mM NaBr for 3HC at pH 3. Both cations and anions strongly influence the SERS enhancement. SERS enhancement depends also significantly on the type of metallic substrates. This indicates the choice of metallic substrate is critically important to achieve strong SERS enhancement. While Ag is the most commonly used materials for SERS substrates and has been demonstrated to exhibit high enhancement. It has the disadvantage of limited selection of excited wavelengths, which prevents to apply Ag SERS substrates to biological field. Dielectric core and metallic shell structure has been theoretically studied and it has been proposed that silica core and silver shell (SiO2@Ag) nanoparticles produces higher plasmon resonance than that of silver nanoparticles and their surface plasmon are tunable by controlling shell thickness. Here, SiO2@Ag nanoparticles were successfully fabricated and their activity as substrates for surface-enhanced Raman scattering (SERS) were examined. Both the core and the shell thickness exhibit strong effect on the SERS activity. Using Rhodamin 6 G (R6G) as a probe molecule, it was observed that SERS intensities of R6G were susceptible to change in Ag shell thickness and the size of core-shell nanoparticles. The 76 nm SiO2@ 23 nm Ag shell nanoparticles shows highest SERS intensity of R6G. Moreover, 76nm SiO2@ 23 nm Ag nanoparticles have higher SERS enhancements of R6G, 4-aminothiophenol (4-ATP), and cotinine (COT) than that of both silver nanoparticles and SiO2@Ag nanoparticles of previous studies. Also, the tuneability of surface plasmon of core-shell structure is flexible by changing in the size of either core or shell. In addition, three Raman spectroscopy application in material science fields were studied: MP-11 encapsulated inside of Tb-mesoMOFs, poly(methyl methacrylate) composites of copper-4,4'-trimethylenedipyridein, and surfactant-free TiO2 surface hydroxyl groups. For the first study, the interaction between the ligands of Tb-meso MOFs and MP-11 was examined. Individual Raman bands of MP-11 and the ligands of Tb-mesoMOFs were distinguished and some of bands were shifted from the complex of MP-11@Tb-mesoMOFs. It is turned out that the interactions is involved through π•••π interactions between the heme and the conjugated triazine and benzene rings of TATB ligand. Next, Raman was used to study the interaction between poly methyl methacrylate (PMMAP) composites and copper-4,4'-trimethylenedipyridein (CU-TMDP). Copper contained in polymer materials has shown improvement performance (thermal and mechanical stability). The Raman results reveal a red-shift of vibrational peaks associated with pyridine ring of CU-TMDP when CU-TMDP is dispersed into PMMA. This interaction, a dipole-dipole interaction or London dispersion force, may produce the stability improvement of metal-containing polymer. The last application is about the effect of pH levels on the phase of TiO2 crystalline. TiO2 crystal has attractive advantage of self-cleaning property. The efficiency of self-cleaning of TiO2 is dependent on the phases (anatase, rutile, and brookite) of TiO2. Raman study revealed that the formation of the anatase phase of TiO2 is interrupted as the pH level increases.

Les études comparatives de la nicotine et de son métabolite obtenu par addition d'une fonction cétone en α par rapport à l'azote du noyau pyrrolidine, la cotinine, ont été réalisées par différentes approches expérimentales : interaction toxique chez la mouche et chez la souris, interaction avec les récepteurs nicotiniques à l'acétylcholine neuronaux et périphériques, étude du passage de la cotinine dans le cerveau et de sa régulation chez le rat, usage de la [125I]cotinine pour étudier les récepteurs de la cotinine, analyse autoradiographique de ces récepteurs, effets de l'administration de nicotine et de cotinine sur la modulation des récepteurs nicotiniques à l'acétylcholine neuronaux et purification du récepteur de la cotinine par chromatographie d'affinité. <br /> Les expériences réalisées, dans les conditions adoptées, ont montré que la métabolisation de la nicotine :<br /><br /> 1 - atténue sa toxicité de 50 fois, par pulvérisation, chez la mouche et de 250 fois, par voie intrapéritoneale, chez la souris.<br /> 2 - donne un produit d'oxydation (cotinine) relativement moins actif et un produit de déméthylation (nornicotine) relativement très actif. Le premier agit avec la nicotine par des mécanismes distincts (supra-additivité ou synergie), alors que le second agit avec la nicotine par des mécanismes communs (additivité ou antagonisme). L'hexaméthonium, un ganglioplégique, ne conférant pas de résistance à la toxicité du mélange de nicotine et de cotinine suggère une toxicité centrale.<br /> 3 - atténue son affinité pour les récepteurs nicotiniques à l'acétylcholine périphériques et neuronaux majeurs.<br /> 4 - atténue fortement son passage dans le cerveau du rat vigilant et soumet ce passage au contrôle du système nicotinique périphérique.<br /> 5 - atténue et modifie ses actions modulatrices sur les récepteurs nicotiniques à l'acétylcholine neuronaux.<br /> 6 - augmente sa sélectivité pour certains récepteurs nicotiniques à l'acétylcholine neuronaux mineurs.<br /><br /> Ainsi, la métabolisation de la nicotine atténue fortement sa toxicité, réduit ses activités et oriente de façon bénéfique ses propriétés pharmacologiques par une augmentation de sa sélectivité pour certains récepteurs nicotiniques à l'acétylcholine neuronaux

No description available.

Diss. (sammanfattning) Stockholm : Karol. inst., 2002. / Härtill 6 uppsatser.

The adverse effects of maternal smoking on infant mortality and morbidity has been well documented in the literature. Maternal tobacco use is causally associated with fetal growth restriction and correlates negatively with folate intake and metabolism. Studies have examined the association between smoking and folate levels during pregnancy, but very few have assessed this relationship using objective and accurate measures of both variables. Furthermore, despite evidence of a causal association between smoking in pregnancy and intrauterine growth restriction, and a plausible relationship between tobacco use and low maternal folate which is required for optimal fetal growth, no experimental study has investigated the potential benefit of folic acid in mitigating the adverse effects of maternal smoking on fetal outcomes. The objectives of this study were to investigate the relationship between maternal smoking and folate levels and examine the efficacy of higher-strength folic acid supplementation, in combination with enrollment in a smoking cessation program, in promoting fetal body and brain growth. Our hypothesis was that women who smoke during pregnancy have lower peri-conceptional folic acid reserves than non-smoker pregnant women and that folic acid reserves will decrease with increasing cotinine level. Additionally, smoker pregnant women on higher-strength folic acid (4mg daily) in combination with smoking cessation programs will experience faster fetal brain growth and have infants with larger body size at birth compared to smokers on the standard dose of folic acid (0.8mg daily). Participants were pregnant women (smokers and non-smokers) who received antenatal care between 2010-2014 at the Genesis Clinic of Tampa, a community health center affiliated with the Department of Obstetrics and Gynecology of the University of South Florida (USF). They were aged 18-44 years and had a gestational age of less than 21 weeks at study enrollment. To determine the peri-conceptional folic acid reserves in smoking versus nonsmoking women during pregnancy and associated sociodemographic factors, baseline (crosssectional) data from a double-blinded randomized controlled trial were analyzed using Tobit regression models (n=496). Smoking information was assessed using salivary cotinine, a sensitive and specific tobacco use biomarker. Folate reserve was measured using red blood cell folate. To investigate the efficacy of higher-strength folic acid on fetal body and brain size, baseline and follow-up data from pregnant smokers enrolled in the randomized controlled trial were utilized (n=345). All primary analyses of the clinical trial data were conducted on a modified intention-to-treat basis and included participants who completed the trial with an observed endpoint, irrespective of compliance to protocol. Multilevel modeling, linear regression, and log-binomial regression analyses were conducted. A significant inverse association between salivary cotinine level and periconceptional red blood cell folate concentration was found among pregnant women in the early to midpregnancy period. Smokers on high-dose folate during pregnancy had infants with a 140.38g higher birth weight than infants of their counterparts on standard dose folate (P =0.047). Mothers who received higher strength folate had a 31.0% lower risk of having babies with SGA compared to their mothers on the standard-dose (adjusted relative risk-ARR=0.69, 95% CI: 0.46–1.03; (P =0.073)). High-dose folate had no significant effect on the intrauterine rate of growth in head circumference, and head circumference and brain weight at birth in our trial sample. However, the brain-body ratio of infants of mothers who received high-dose treatment was 0.33 percentage-point lower than that for infants of mothers who received the standard dose of folate (P =0.044). Higher strength folic acid supplementation in pregnant women who smoke might be a cost-effective and safe option to improve birth outcomes and reduce low birth weight and SGA associated infant morbidity and mortality. Future studies with larger sample sizes and diverse populations are indicated to confirm or refute the results of this study. Randomized controlled trials starting during the preconception period and with follow-up until delivery are warranted, to identify the most folate-sensitive period of fetal growth and determine the optimal dose of folic acid supplement. Further research investigating several pathways through which the effects of prenatal smoking on adverse birth outcomes can be mitigated is needed.

Introdução: Muitos estudos relacionam o tabagismo materno durante a gestação com o aumento do risco de desfechos adversos para o feto, como o baixo peso ao nascer e restrição de crescimento intrauterino. Os objetivos deste trabalho foram: estabelecer um modelo animal de exposição ao tabaco durante a prenhez, usando como marcador de exposição a cotinina e a carboxihemoglobina; investigar os efeitos da exposição pré-natal ao tabaco sobre a biometria fetal e parâmetros bioquímicos; e investigar a programação fetal e suas repercussões na vida adulta. Métodos: Ratas Wistar foram randomicamente divididas em 3 grupos de exposição ao tabaco durante a gestação: controle (C), controle manipulado (CM) e tabaco (T). No grupo T as ratas foram expostas a fumaça de um cigarro 2x/dia, durante 21 dias. O grupo CM passou pela mesma intervenção do grupo T, porém sem sofrer exposição à fumaça do cigarro. O grupo C permaneceu na caixa-moradia sem qualquer tipo de interferência. O protocolo do presente estudo foi dividido em 2 experimentos. Experimento 1: no 22º dia de gestação um grupo de ratas genitoras foi submetida à cirurgia cesariana para a retirada dos filhotes. O sangue do tronco das genitoras foi coletado e o peso da placenta, bem como o comprimento e o peso dos filhotes foram medidos. Experimento 2: no 22º dia de gestação, um outro grupo de genitoras tiveram seus filhotes nascidos de parto normal e as ninhadas foram padronizadas em 8 filhotes. O peso dos filhotes foi aferido semanalmente até a vida adulta, quando foram realizados o teste de tolerância à glicose, o teste de corticosterona em resposta ao estresse e a medida da gordura abdominal. Para a medida da carboxihemoglobina (COHb) nas genitoras o sangue foi coletado imediatamente após a exposição a fumaça do cigarro. Resultados: O grupo exposto ao tabaco de forma aguda apresentou níveis mais elevados de COHb que o grupo CM. A exposição repetida ao tabaco aumentou substancialmente os níveis de cotinina, enquanto os grupos CM e C apresentaram níveis de cotinina abaixo do limite de detecção do método. Foi observado que os filhotes expostos ao tabaco no período intrauterino nasceram com menor peso e menores níveis de insulina e glicemia. Em relação ao experimento 2, o acompanhamento do ganho de peso até a vida adulta não diferiu entre os grupos, somente entre os sexos, assim como a adiposidade. O teste de tolerância à glicose na vida adulta mostrou que o grupo exposto ao tabaco durante o período intrauterino apresentou resistência à ação da insulina. Conclusão: O modelo animal utilizado para expor ao tabaco durante a prenhez foi capaz de induzir a restrição do crescimento intrauterino (RCIU) e de programar o metabolismo na vida adulta. Este modelo animal mostra-se útil na investigação de outros desfechos e mecanismos relacionados à exposição tabágica durante a gestação. / Introduction: Many studies have linked maternal smoking during pregnancy with increased risk of adverse outcomes. The objective of this study was to establish an animal model of tobacco exposure during pregnancy, using as a marker of exposure to cotinine and carboxyhemoglobin. Also, investigate the effects of prenatal exposure to tobacco smoke on fetal biometry and biochemical parameters. It also aimed to investigate the fetal programming and its repercussions in adulthood. Methodology: Repeated exposure to tobacco smoke during pregnancy: The animals were randomly assigned to three groups: control (C), manipulated control (MC) and tobacco (T). The animals were exposed to a one cigarette twice a day for 21 days. The manipulated control group went through the same intervention group T, without suffering smoke exposure. The group C remained intact in the home cage. Experiment 1: on 22nd day of pregnancy was performed a cesarean section. The trunk blood was collected of the pregnant rats and fetal biometry measured. Experiment 2: on the 22nd day of pregnancy the pups were born by natural delivery and the litters were standardized to eight pups per litter. The weight of offspring was measured weekly until adulthood. At this stage we evaluated the glucose tolerance test and, at the time of sacrifice, the adiposity. Immediately after the intervention, the trunk blood was collected for carboxyhemoglobin (COHb) measurement. Results: the animals exposed to tobacco acutely showed higher levels of COHb that the MC group. Repeated exposure to tobacco increased substantially cotinine levels, whereas in MC and C groups cotinine levels were below the limit of detection. It was observed that pups exposed to tobacco intrauterine period were born with lower weight and lower levels of insulin and glucose. Regarding the second experiment, monitoring of weight gain until adulthood did not differ between groups, only gender, and adiposity. The glucose tolerance test in adult life showed that the group exposed to tobacco during the intrauterine period showed resistance to insulin action. Conclusion: The animal model used to expose the tobacco during pregnancy was able to induce intrauterine growth restriction and programming metabolism in later life. This animal model shown to be useful for studies on other outcomes, and mechanisms related to tobacco smoke exposure during pregnancy.

Introdução: Muitos estudos relacionam o tabagismo materno durante a gestação com o aumento do risco de desfechos adversos para o feto, como o baixo peso ao nascer e restrição de crescimento intrauterino. Os objetivos deste trabalho foram: estabelecer um modelo animal de exposição ao tabaco durante a prenhez, usando como marcador de exposição a cotinina e a carboxihemoglobina; investigar os efeitos da exposição pré-natal ao tabaco sobre a biometria fetal e parâmetros bioquímicos; e investigar a programação fetal e suas repercussões na vida adulta. Métodos: Ratas Wistar foram randomicamente divididas em 3 grupos de exposição ao tabaco durante a gestação: controle (C), controle manipulado (CM) e tabaco (T). No grupo T as ratas foram expostas a fumaça de um cigarro 2x/dia, durante 21 dias. O grupo CM passou pela mesma intervenção do grupo T, porém sem sofrer exposição à fumaça do cigarro. O grupo C permaneceu na caixa-moradia sem qualquer tipo de interferência. O protocolo do presente estudo foi dividido em 2 experimentos. Experimento 1: no 22º dia de gestação um grupo de ratas genitoras foi submetida à cirurgia cesariana para a retirada dos filhotes. O sangue do tronco das genitoras foi coletado e o peso da placenta, bem como o comprimento e o peso dos filhotes foram medidos. Experimento 2: no 22º dia de gestação, um outro grupo de genitoras tiveram seus filhotes nascidos de parto normal e as ninhadas foram padronizadas em 8 filhotes. O peso dos filhotes foi aferido semanalmente até a vida adulta, quando foram realizados o teste de tolerância à glicose, o teste de corticosterona em resposta ao estresse e a medida da gordura abdominal. Para a medida da carboxihemoglobina (COHb) nas genitoras o sangue foi coletado imediatamente após a exposição a fumaça do cigarro. Resultados: O grupo exposto ao tabaco de forma aguda apresentou níveis mais elevados de COHb que o grupo CM. A exposição repetida ao tabaco aumentou substancialmente os níveis de cotinina, enquanto os grupos CM e C apresentaram níveis de cotinina abaixo do limite de detecção do método. Foi observado que os filhotes expostos ao tabaco no período intrauterino nasceram com menor peso e menores níveis de insulina e glicemia. Em relação ao experimento 2, o acompanhamento do ganho de peso até a vida adulta não diferiu entre os grupos, somente entre os sexos, assim como a adiposidade. O teste de tolerância à glicose na vida adulta mostrou que o grupo exposto ao tabaco durante o período intrauterino apresentou resistência à ação da insulina. Conclusão: O modelo animal utilizado para expor ao tabaco durante a prenhez foi capaz de induzir a restrição do crescimento intrauterino (RCIU) e de programar o metabolismo na vida adulta. Este modelo animal mostra-se útil na investigação de outros desfechos e mecanismos relacionados à exposição tabágica durante a gestação. / Introduction: Many studies have linked maternal smoking during pregnancy with increased risk of adverse outcomes. The objective of this study was to establish an animal model of tobacco exposure during pregnancy, using as a marker of exposure to cotinine and carboxyhemoglobin. Also, investigate the effects of prenatal exposure to tobacco smoke on fetal biometry and biochemical parameters. It also aimed to investigate the fetal programming and its repercussions in adulthood. Methodology: Repeated exposure to tobacco smoke during pregnancy: The animals were randomly assigned to three groups: control (C), manipulated control (MC) and tobacco (T). The animals were exposed to a one cigarette twice a day for 21 days. The manipulated control group went through the same intervention group T, without suffering smoke exposure. The group C remained intact in the home cage. Experiment 1: on 22nd day of pregnancy was performed a cesarean section. The trunk blood was collected of the pregnant rats and fetal biometry measured. Experiment 2: on the 22nd day of pregnancy the pups were born by natural delivery and the litters were standardized to eight pups per litter. The weight of offspring was measured weekly until adulthood. At this stage we evaluated the glucose tolerance test and, at the time of sacrifice, the adiposity. Immediately after the intervention, the trunk blood was collected for carboxyhemoglobin (COHb) measurement. Results: the animals exposed to tobacco acutely showed higher levels of COHb that the MC group. Repeated exposure to tobacco increased substantially cotinine levels, whereas in MC and C groups cotinine levels were below the limit of detection. It was observed that pups exposed to tobacco intrauterine period were born with lower weight and lower levels of insulin and glucose. Regarding the second experiment, monitoring of weight gain until adulthood did not differ between groups, only gender, and adiposity. The glucose tolerance test in adult life showed that the group exposed to tobacco during the intrauterine period showed resistance to insulin action. Conclusion: The animal model used to expose the tobacco during pregnancy was able to induce intrauterine growth restriction and programming metabolism in later life. This animal model shown to be useful for studies on other outcomes, and mechanisms related to tobacco smoke exposure during pregnancy.

Introdução: Muitos estudos relacionam o tabagismo materno durante a gestação com o aumento do risco de desfechos adversos para o feto, como o baixo peso ao nascer e restrição de crescimento intrauterino. Os objetivos deste trabalho foram: estabelecer um modelo animal de exposição ao tabaco durante a prenhez, usando como marcador de exposição a cotinina e a carboxihemoglobina; investigar os efeitos da exposição pré-natal ao tabaco sobre a biometria fetal e parâmetros bioquímicos; e investigar a programação fetal e suas repercussões na vida adulta. Métodos: Ratas Wistar foram randomicamente divididas em 3 grupos de exposição ao tabaco durante a gestação: controle (C), controle manipulado (CM) e tabaco (T). No grupo T as ratas foram expostas a fumaça de um cigarro 2x/dia, durante 21 dias. O grupo CM passou pela mesma intervenção do grupo T, porém sem sofrer exposição à fumaça do cigarro. O grupo C permaneceu na caixa-moradia sem qualquer tipo de interferência. O protocolo do presente estudo foi dividido em 2 experimentos. Experimento 1: no 22º dia de gestação um grupo de ratas genitoras foi submetida à cirurgia cesariana para a retirada dos filhotes. O sangue do tronco das genitoras foi coletado e o peso da placenta, bem como o comprimento e o peso dos filhotes foram medidos. Experimento 2: no 22º dia de gestação, um outro grupo de genitoras tiveram seus filhotes nascidos de parto normal e as ninhadas foram padronizadas em 8 filhotes. O peso dos filhotes foi aferido semanalmente até a vida adulta, quando foram realizados o teste de tolerância à glicose, o teste de corticosterona em resposta ao estresse e a medida da gordura abdominal. Para a medida da carboxihemoglobina (COHb) nas genitoras o sangue foi coletado imediatamente após a exposição a fumaça do cigarro. Resultados: O grupo exposto ao tabaco de forma aguda apresentou níveis mais elevados de COHb que o grupo CM. A exposição repetida ao tabaco aumentou substancialmente os níveis de cotinina, enquanto os grupos CM e C apresentaram níveis de cotinina abaixo do limite de detecção do método. Foi observado que os filhotes expostos ao tabaco no período intrauterino nasceram com menor peso e menores níveis de insulina e glicemia. Em relação ao experimento 2, o acompanhamento do ganho de peso até a vida adulta não diferiu entre os grupos, somente entre os sexos, assim como a adiposidade. O teste de tolerância à glicose na vida adulta mostrou que o grupo exposto ao tabaco durante o período intrauterino apresentou resistência à ação da insulina. Conclusão: O modelo animal utilizado para expor ao tabaco durante a prenhez foi capaz de induzir a restrição do crescimento intrauterino (RCIU) e de programar o metabolismo na vida adulta. Este modelo animal mostra-se útil na investigação de outros desfechos e mecanismos relacionados à exposição tabágica durante a gestação. / Introduction: Many studies have linked maternal smoking during pregnancy with increased risk of adverse outcomes. The objective of this study was to establish an animal model of tobacco exposure during pregnancy, using as a marker of exposure to cotinine and carboxyhemoglobin. Also, investigate the effects of prenatal exposure to tobacco smoke on fetal biometry and biochemical parameters. It also aimed to investigate the fetal programming and its repercussions in adulthood. Methodology: Repeated exposure to tobacco smoke during pregnancy: The animals were randomly assigned to three groups: control (C), manipulated control (MC) and tobacco (T). The animals were exposed to a one cigarette twice a day for 21 days. The manipulated control group went through the same intervention group T, without suffering smoke exposure. The group C remained intact in the home cage. Experiment 1: on 22nd day of pregnancy was performed a cesarean section. The trunk blood was collected of the pregnant rats and fetal biometry measured. Experiment 2: on the 22nd day of pregnancy the pups were born by natural delivery and the litters were standardized to eight pups per litter. The weight of offspring was measured weekly until adulthood. At this stage we evaluated the glucose tolerance test and, at the time of sacrifice, the adiposity. Immediately after the intervention, the trunk blood was collected for carboxyhemoglobin (COHb) measurement. Results: the animals exposed to tobacco acutely showed higher levels of COHb that the MC group. Repeated exposure to tobacco increased substantially cotinine levels, whereas in MC and C groups cotinine levels were below the limit of detection. It was observed that pups exposed to tobacco intrauterine period were born with lower weight and lower levels of insulin and glucose. Regarding the second experiment, monitoring of weight gain until adulthood did not differ between groups, only gender, and adiposity. The glucose tolerance test in adult life showed that the group exposed to tobacco during the intrauterine period showed resistance to insulin action. Conclusion: The animal model used to expose the tobacco during pregnancy was able to induce intrauterine growth restriction and programming metabolism in later life. This animal model shown to be useful for studies on other outcomes, and mechanisms related to tobacco smoke exposure during pregnancy.