Orientador: José Antonio Rocha Gontijo / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T06:22:30Z (GMT). No. of bitstreams: 1 Possignolo_LuizFernando_M.pdf: 4767580 bytes, checksum: b843458e4c7049fc29741e4aca9f918d (MD5) Previous issue date: 2012 / Resumo: Devido ao maior consumo de alimentos ricos em gordura e um estilo de vida mais sedentário, houve um aumento na incidência de desordens metabólicas relacionadas ao metabolismo lipídico como a resistência à insulina, dislipidemias, e sua associação com doenças cardiovasculares. A alimentação materna desequilibrada, durante a gestação e lactação, pode predispor a prole à doenças durante a vida adulta. Alguns transportadores como o Scavenger Receptor class B type I (SR-BI) e ATP-binding cassette transporter A1 (ABCA1) são descritos como responsáveis pela captação de colesterol e transporte deste a partir de lipoproteínas, principalmente no transporte reverso de colesterol, sendo denominados receptores antiaterogênicos podendo modificar seu padrão de expressão frente a uma dieta hiperlipídica. Ratas Wistar receberam dieta hiperlipídica (DHL) ou dieta padrão (CTL) desde o desmame, até a lactação. Após o desmame a prole de machos recebeu a dieta padrão até a 16ª semana de vida. Nas mães quanto e na prole foram analisados os seguintes parâmetros: ingestão alimentar, ganho ponderal, perfil lipídico e glicídico. Na prole foi estudada a expressão e localização de ABCA1 e SR-BI na placenta, no rim, fígado, e intestino delgado em animais com 17 dias pré-natal (E17), 12 dias pós-natal (PN12d) com 8 e 16 semanas pós-natal (PN8s e PN16s). As mães DHL apresentaram: 1) maior ingestão calórica com menor ganho ponderal; 2) aumento glicêmico associado à menor produção de insulina nos três períodos estudados e, 3) aumento nos níveis séricos de triglicérides em M8s. A prole de mães DHL apresentaram menor massa corporal desde E17 até PN8s, sem que tenha havido diferenças ponderais e na ingestão de ração. PN8s e PN16S apresentaram menor captação tissular de glicose associada à hiperinsulinemia, e aumento nos níveis séricos de triglicérides com PN16S. Não houve alterações nos níveis de colesterol e HDLcolesterol. Não foi observada alteração na expressão de SR-BI e ABCA1 no intestino delgado, placenta enquanto no fígado houve uma queda tempo-dependente para ambos transportadores. No rim da prole DHL aos PN12d e PN16s observou-se maior expressão de ABCA1. Este estudo mostrou que o consumo materno crônico de uma dieta hiperlipídica causa alterações metabólicas nas mães e predispõe a prole, a modificações no metabolismo lipídico e glicídico, além de elevação da expressão de ABCA1 no rim / Abstract: Due to the abundance and accessibility to foods high in fat and a more sedentary lifestyle, there is an increased incidence of metabolic disorders related to lipid metabolism such as insulin resistance, dyslipidemia, and a high correlation with cardiovascular disease. The unbalanced maternal diet during pregnancy and lactation predisposes the offspring to diseases during adult life. Some carriers such as scavenger receptor class B type I (SR-BI) and ATP-binding cassette transporter A1 (ABCA1) are described as responsible for raising cholesterol and transporting it to and from lipoproteins, due the participation in the reverse cholesterol transport, these receptors are called antiatherogenic and are subject to change its pattern of expression when exposed to a high fat diet. Female Wistar rats were fed a diet (HFD) or a standard chow (CTL) from weaning, during pregnancy and lactation. After weaning the male offspring was exposed to a standard chow until the 16th week of life. Both the dams and the offsprings food intake were monitored, weight gain, lipid profile and glucose level. It was analyzed in the offspring the expression and localization of ABCA1 and SR-BI in the placenta, kidney, liver, and small intestine in animals at 17º prenatal day (E17), 12 days post-natal (PN12d), 8 and 16 postnatal weeks (PN8s PN16s respectively). DHL dams had a higher intake of calories in the diet, but the weight was smaller, they had higher blood glucose due to decreased production of insulin in the pre-pregnancy (m8s), during pregnancy (M17g) and lactation (M15l) and a higher triglyceride level in m8s. The offspring of dam fed a high fat diet had lower weight since E17 until PN8s, with no differences in weight gain and food intake. PN8s and PN16s had lower glucose uptake and hyperinsulinemia, and high triglycerides with PN16s, no changes were observed in cholesterol and HDL-cholesterol levels. There were no changes in the expression of SR-BI and ABCA1 in the small intestine, placenta and liver, however there was a decrease over the age for both receptors, and kidney of the offspring and DHL PN12d PN16s showed higher expression of ABCA1. The present study showed that chronic consumption of high fat diet causes metabolic changes in dams and predisposes offspring to changes in lipid and glucose metabolism of the offspring, increasing the expression of ABCA1 in the kidney / Mestrado / Biologia Estrutural, Celular, Molecular e do Desenvolvimento / Mestre em Fisiopatologia Médica

Orientadores: Cleisson Fábio Andrioli Peralta, Ricardo Barini / Dissertação (Mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-20T20:27:17Z (GMT). No. of bitstreams: 1 Ferreira_SabrinaGirotto_M.pdf: 3220177 bytes, checksum: fc5df188c73f1cc85083481b3efdcf1b (MD5) Previous issue date: 2012 / Resumo: Objetivo: Estabelecer intervalos de referência de idade gestacional (IG), índices de pulsatilidade (IP) das artérias umbilical (AU) e média das artérias uterinas (AUT) de acordo com o grau placentário de Grannum. Esses dados servirão como base para estudo em andamento que avalia o impacto do amadurecimento precoce da placenta nos parâmetros dopplervelocimétricos acima mencionados e os resultados perinatais. Sujeitos e métodos: estudo prospectivo longitudinal observacional realizado em hospital universitário terciário, em que 133 gestações de baixo risco foram avaliadas quinzenalmente pela USG, entre 18 e 41 semanas. A classificação placentária de Grannum et al. (1), IP-AU e IPm-AUT foram obtidos em cada exame. Os intervalos de referência (mediana, 5?, 10?, 90? e 95? percentil) de IG, IP-AU e IPm-AUT foram estabelecidos para cada grau placentário. Testes de Mann-Whitney com correção de Bonferroni foram utilizados para comparar os parâmetros acima mencionados entre dois diferentes graus placentários. O valor de p bicaudal menor que 0,05 foi considerado estatisticamente significativo. Resultados: A IG aumentou significativamente com a mudança do grau placentário. IP-AU e IP-AUT reduziram significativamente do grau zero para o um e do grau um para o dois, mas permaneceram estáveis depois disso. Conclusões: Os intervalos de referência dos parâmetros dopplervelocimétricos que refletem implantação normal e função placentária foram elaborados como um primeiro passo que permitirá comparar e testar nossos dados com as publicações existentes para a predição de resultados perinatais em casos de amadurecimento precoce da placenta em gestações de baixo risco / Abstract: Objective: To establish reference intervals of gestational age (GA), umbilical artery (UmbA) and mean uterine artery (mUtA) Doppler pulsatility indexes (PI) according to placental grade. This will serve as a basis for an ongoing study aimed at evaluating the impact of early placental ageing and the abovementioned Doppler parameters on pregnancy outcomes. Materials and Methods: Longitudinal observational study realized in tertiary university hospital where 133 low-risk pregnancies were scanned fortnightly from 18 to 41 weeks. Placental classification according to Grannum et al. (1), UmbA-PI and and mUtA-PI were obtained in each scan. Reference intervals (median, 5th, 10th, 90th and 95th centiles) of GA, UmbA-PI and mUtA-PI were established for each placental grade. Mann-Whitney U tests with Bonferroni adjustments were used to compare these parameters between two different placental grades. Two-tailed p-values of less than 0.05 were considered statistically significant. Results: GA significantly increased as placental grade changed. UmbA-PI and mUtA-PI significantly decreased between grades zero and one and between grades one and two, but remained stable between grades two and three. Conclusions: Reference intervals of GA and Doppler parameters which reflect normal implantation and function of the placenta were elaborated as a first step to allow further testing and comparison of our data with those previously published for the prediction of pregnancy outcomes in cases of suspicion of an early placental ageing is suspected / Mestrado / Saúde Materna e Perinatal / Mestre em Ciências da Saúde

Objective: To determine the prevalence and odds ratios for cleft lip and/or palate (CL/P) among infants prenatally exposed to opioids with or without neonatal opioid withdrawal syndrome (NOWS). Design: This study represents an exploratory, retrospective cohort study design of newborn medical health records from 2011 to 2016. Setting: Records were drawn from a regional health system located in South Central Appalachia. Population and Study Sample: The original population yielded 3 cohorts of infants: (1) infants with opioid exposure (OE) but not requiring pharmacological intervention (OE; N = 168); (2) infants with NOWS requiring pharmacological intervention (N = 294); and (3) infants with no opioid exposure (NOE; N = 16 090), the primary comparison group. Main Outcome: Infants in the NOWS and OE groups showed significantly increased prevalence and odds ratios for CL/P when compared to those in the NOE group. Results: Prevalence rates per 1000 live births for infants with OE (35.71) and infants with NOWS (6.80) were significantly higher than those for infants with NOE (1.37). Comparison of infants with OE to the NOE group revealed significantly increased odds for CL/P, isolated cleft palate (CP), cleft lip (CL), and cleft lip and palate (CLP) (27.05, 41.81, 19.26, 19.37, respectively; all Ps <.008). The odds ratios for infants with NOWS compared to the NOE group were significantly higher for CL/P and CP (5.00 and 10.98, respectively; Ps <.03) but not for CL and CLP. Conclusion: The results provide additional evidence that prenatal OE should be considered among the critical environmental risk factors that can contribute to CL/P.

Preterminfants frequently suffer frompulmonary complications due to a physiological and structural lung immaturity resulting in significant morbidity and mortality. Novel in vitro and in vivo models are required to study the underlying mechanisms of late lung maturation and to facilitate the development of new therapeutic strategies. Organoids recapitulate essential aspects of structural organization and possibly organ function, and can be used to model developmental and disease processes. We aimed at generating fetal lung organoids (LOs) and to functionally characterize this in vitro model in comparison to primary lung epithelial cells and lung explants ex vivo. LOs were generated with alveolar and endothelial cells from fetal rat lung tissue, using a Matrigel-gradient and air-liquid-interface culture conditions. Immunocytochemical analysis showed that the LOs consisted of polarized epithelial cell adhesion molecule (EpCAM)-positive cells with the apical membrane compartment facing the organoid lumen. Expression of the alveolar type 2 cell marker, RT2-70, and the Club cell marker, CC-10, were observed. Na+ transporter and surfactant protein mRNA expression were detected in the LOs. First time patch clamp analyses demonstrated the presence of several ion channels with specific electrophysiological properties, comparable to vital lung slices. Furthermore, the responsiveness of LOs to glucocorticoids was demonstrated. Finally, maturation of LOs induced by mesenchymal stem cells confirmed the convenience of the model to test and establish novel therapeutic strategies. The results showed that fetal LOs replicate key biological lung functions essential for lung maturation and therefore constitute a suitable in vitro model system to study lung development and related diseases.

<p dir="ltr">Epigenetics involves the study of mechanisms that influence gene expression. These mechanisms are heritable and dynamic, and despite altering gene transcriptional activity, they do not change the underlying DNA sequence. While epigenetic mechanisms govern gene expression throughout the lifetime of an organism, the dynamic nature and precision of the transcriptional control afforded by processes such as histone modifications and chromatin architecture remodeling are exemplified in early mammalian development. Perhaps unsurprisingly, perturbations to the epigenetic status of a cell can alter its function, and widespread epigenome disruptions due to changes in the developmental environment can compromise the growth, and even viability, of an embryo or fetus. By studying epigenetic mechanisms and the patterns they impart, we can better understand not only how developmental progression is regulated during embryonic development and beyond, but also what the consequences of aberrant epigenetic disturbances may be to developing organisms.</p><p><br></p><p dir="ltr">Many gaps remain in our knowledge concerning epigenetic mechanisms in domestic livestock species, particularly regarding early development. Pigs represent a compelling model organism for study in this area, as they are increasingly being used as a biomedical model for human-oriented research due to their physiological similarities to humans, and they remain a staple meat species in many parts of the world. Chapter 2 investigates the presence and transcriptional dynamics of the SWI/SNF chromatin remodeling complex GBAF in porcine trophectoderm (PTr2) and fetal fibroblast (PFF) cells. These cell lines represent two discrete developmental stages during early swine development, with the PTr2 cells originally obtained from the trophectoderm of a gestation day 12 elongating porcine conceptus, and the fetal fibroblast cells were collected from a fetus on day 40 of gestation. Using immunocytochemistry and Western blotting techniques, GBAF was identified in both cell lines. Further, co-immunoprecipitation of GBAF constituent subunits and other BAF family subcomplex subunits revealed a previously undescribed interaction between the GBAF subunit GLTSCR1 and the BAF subunit BAF170, the latter of which has not been shown to be present in human and mouse GBAF. This may suggest a species-specific GBAF composition in swine. Analysis of RNA-seq data from porcine embryos, PTr2 cells, and PFF cells showed that while transcription of GBAF-specific subunits BRD9 and GLTSCR1 was detectable, expression levels were lower compared to other BAF family subunits. Taken together these results suggest that, while GBAF is detectable in swine early development contexts, it may have a comparatively minor contribution as an epigenetic mechanism during the represented developmental timepoints.</p><p><br></p><p dir="ltr">Details in the literature about the epigenetic landscape and the resulting chromatin state during porcine early development are also limited at present. Chapter 3 involves the global epigenetic profiling of the histone marks H3K4me3, H3K27ac, and H3K27me3 and the SWI/SNF central ATPase BRG1 in PTr2 and PFF cells using CUT&RUN. The enrichment patterns observed for these features were consistent with known patterns described in the literature. H3K4me3 was primarily enriched in gene promoter regions, and H3K27ac showed enrichment in both promoter regions and distal intergenic regions, some of which are likely active enhancers. H3K27me3 showed broad genomic localization and was detected at genes known to be transcriptionally inactive in these cell types, as well as in distal intergenic regions. BRG1 showed some co-enrichment with H3K4me3 and H3K27ac in promoter regions, as well as several instances of H3K27ac co-enrichment at intergenic sites. The sequencing files were used to build a chromatin state prediction model for 10 states in each cell line, ranging from TSS to repressed genomic regions. Additionally, the transcriptomes of PTr2 and PFF cells were compared to those of human cells taken from comparable gestational time points to determine if these swine cell lines could potentially serve as translational <i>in vitro</i> models. PTr2 cells and human trophectoderm (TE) cells were relatively dissimilar in their cell-type specific gene identities (~24% overlap) and corresponding transcriptional levels, but the porcine and human fibroblast cells shared around 50% of the same cell type-specific genes, and expression levels were broadly similar among them. Altogether, these findings provide foundational epigenetic landscape information for PTr2 and PFF cells and potential insights regarding similarities and differences in cell identity between human and pig trophectoderm and fetal fibroblast cells.</p><p><br></p><p dir="ltr">The placenta is a transient organ that provides essential support to the developing fetus in the form of nutrient and gas exchange. Despite its significance in facilitating fetal development, our understanding of how the placenta is affected by its environment is greatly limited, and only a handful of studies exploring the placental epigenome in swine exist to date. To address these gaps, and building upon the epigenetic profiling methods developed in Chapter 3, Chapter 4 investigated whether, and to what extent, the placental epigenome changes in response to fetal endocrine perturbations. Placental tissue was collected at day 86 of gestation from untreated pregnant gilts and pregnant gilts treated for 21 days with methimazole (MMI) to induce fetal hypothyroidism. CUT&RUN was used to evaluate the enrichment of H3K4me3, H3K27ac, and BRG1 in placental tissue derived from n=6 male and female fetuses in each treatment group (n=12 samples per group). Differential enrichment of all three epigenetic features was seen in placental tissues obtained from MMI-treated fetuses, and, notably, existing sex-specific differences in placental epigenetic features were exacerbated by MMI-induced fetal hypothyroidism. This may suggest that the porcine placenta may be impacted by fetal endocrine status during late gestation. Together, these findings show that sex-specific differences in placental chromatin state exist and that a fetal hypothyroid state is sufficient to perturb the placental epigenome, ultimately providing novel insights into the intricate interplay between fetal endocrine status and regulation of the placental epigenome.</p>

Estudos em animais mostram que a desnutrição proteica intrauterina pode acarretar redistribuição do fluxo sanguíneo intraútero, podendo promover modificações permanentes na estrutura e funcionalidade de alguns órgãos, o que ocasiona modificações no metabolismo. Além disso, a desnutrição intrauterina pode afetar a secreção de hormônios que atuam no crescimento fetal, podendo conduzir à restrição do crescimento intrauterino. Esse fenômeno pode parcialmente ser explicado pela hipótese da programação fetal, na qual é sugerido que ocorra uma adaptação metabólica e fisiológica do feto a uma condição intrauterina adversa, que pode induzir o desenvolvimento de doenças crônicas não transmissíveis na vida adulta. Neste contexto, pesquisas com suplementação de aminoácidos de cadeia ramificada (BCAA) têm verificado a capacidade desses nutrientes promoverem a síntese proteica mesmo em condições catabólicas, por meio da ativação de uma via bioquímica intracelular intercedida pela proteína quinase Alvo da Rapamicina em Mamíferos (mTOR), a qual está envolvida no estímulo à etapa de tradução proteica. Assim, o presente trabalho avaliou o efeito da suplementação de BCAA em proles submetidas à desnutrição proteica materna. Para tanto, ratas Wistar foram acasaladas com ratos adultos de mesma raça. Uma vez constatada a gravidez, as matrizes foram distribuídas em grupos de acordo com a dieta que seria fornecida no decorrer da gestação: CON (20% proteína); VAL/ISO (5% proteína + 2% VAL + 2% ISO); AAE (5% proteína + 4% AAE); e BCAA (5% proteína + 4% BCAA). O protocolo de restrição proteica materna adotado causou redução no crescimento corporal e na massa de órgãos das proles. Embora a suplementação com VAL/ISO e AAE não tenha recuperado os efeitos mediados pela deficiência de proteína, foi constatado que a suplementação com BCAA reverteu parte do déficit observado no crescimento das proles, uma vez que foi eficaz em minimizar ou mesmo em restaurar plenamente diversos parâmetros como peso de órgãos, massa de gordura da carcaça e parâmetros indicativos do estado nutricional proteico, como as concentrações de proteína e RNA hepáticas e musculares. Estes efeitos podem parcialmente ser explicados pelo estímulo induzido pela suplementação com BCAA, na via de sinalização da mTOR, considerando que foi verificado no fígado das proles de matrizes que receberam esta suplementação, aumento na fosforilação desta proteína (P < 0,05), a qual é responsável por desencadear uma cascata de eventos biomoleculares que culminam, em última instância, no acréscimo da síntese proteica. Diante disto, torna-se relevante a realização de pesquisas que avaliem em longo prazo, os efeitos da suplementação com BCAA em proles submetidas à dieta materna restrita em proteína. / Animal studies show that intrauterine malnutrition may cause redistribution of blood flow in uterus, which may promote permanent changes in structure and function of some organs, which causes changes in metabolism. Furthermore, intrauterine malnutrition can affect the secretion of hormones that act on fetal growth and may lead to intrauterine growth restriction. This phenomenon can partly be explained by the hypothesis of fetal programming, which is suggested that occur a metabolic and physiological adaptation of the fetus to an adverse intrauterine condition, which can induce the development of chronic diseases in later life. In this context, researches with supplementation of branched chain amino acids (BCAA), especially leucine, have verified the ability of these nutrients to promote protein synthesis in catabolic conditions, through the activation of an intracellular biochemical pathway interceded by protein kinase Mammalian Target of Rapamycin (mTOR), which is involved in the stimulating of protein translation stage. Thus, this study evaluated the effect of BCAA supplementation in offspring subjected to maternal protein-restrict diet. To this, Wistar rats were mated with adult rats of the same race. Once was confirmed the pregnancy, the pregnants were distributed into groups according to the diet that would be provided during pregnancy: CON (20% protein); VAL/ISO (5% protein + 2% + 2% VAL/ISO), AAE (5% protein + 4% EAA) and BCAA (5% protein + 4% BCAA). The protocol adopted maternal protein restriction caused a reduction in body growth and weight of the offspring\'s organs. Although supplementation with VAL/ISO and AAE has not recovered the effects mediated by protein deficiency, it was found that supplementation with BCAA has reversed part of the deficit observed in the growth of the offspring, since it was effective in minimizing or even fully restoring various parameters such as organ weight, carcass fat mass and parameters indicative of nutritional protein, such as the concentrations of protein and RNA in liver and muscle. These effects may be partially explained by the stimulation induced by BCAA supplementation on the mTOR signaling pathway, considering that was verified in the liver of the offspring from dams that received this supplementation augment on the phosphorylation of this protein (P < 0,05), which is responsible for triggering a cascade of molecular events that culminate, ultimately, in increased protein synthesis. Given this, it becomes relevant to conducting research to assess long-term effects of supplementation with BCAA in offspring subjected to maternal protein-restricted diet.

Introdução: O Diabetes mellitus tipo 1 (DM1) cursa com produção ausente ou irrisória de insulina e é a forma responsável pelos casos mais graves de distúrbios glicêmicos. O DM1 exerce forte influência sobre o crescimento fetal. Enquanto a hiperglicemia estimula o crescimento fetal devido à hiperinsulinemia, nas pacientes com vasculopatias a placentação inadequada pode levar o feto à restrição. Objetivos: identificar alterações de crescimento fetal e avaliá-las quanto a sua associação com o controle glicêmico materno e de marcadores laboratoriais de função renal. Métodos: foram avaliadas, de forma prospectiva em coorte observacional, 60 gestantes com DM1 que iniciaram o pré-natal no primeiro trimestre. A associação entre a classificação de peso ao nascimento com as seguintes variáveis foi analisada: média glicêmica, frequência de hipo e hiperglicemia, frequência de hipo e hiperglicemia grave, hemoglobina glicada, frutosamina, ácido úrico, creatinina e proteinúria de 24 horas. A predição do crescimento fetal também foi estudada. Resultados: Desvios do crescimento fetal em pacientes com DM1 ocorreram em 41% dos casos (n=25). Observou-se que 10% das gestações resultaram em PIG (n=6) e 31%, em GIG (n=19). Níveis aumentados de média glicêmica (p =0,006), baixa frequência de hipoglicemias (p = 0,027) e alta frequência de hiperglicemias (p = 0,014) se associaram a GIG no terceiro trimestre. Em todos os trimestres, valores séricos mais elevados de ácido úrico, creatinina e proteinúria de 24hs, se associaram de maneira significativa, ao grupo PIG. Foi construído um modelo, com taxa de acerto de 80.3%, para a predição de crescimento fetal com os valores de terceiro trimestre da média glicêmica e da creatinina. Conclusões: Foram identificadas variáveis relacionadas ao controle glicêmico materno e à marcadores laboratoriais de função renal que se associaram a alterações no crescimento fetal. Usando algumas dessas variáveis foi possível construir um modelo para predição do crescimento fetal com boa acurácia / Introduction. Diabetes mellitus type 1 (DM1) is described as absent or negligible production of insulin and it is responsible for the most severe cases of glycemic disorders. DM1 has a strong influence on fetal growth. In pregnant women the hyperglycemia stimulates fetal growth, and the vasculopathy influences the placentation process, which may lead to growth restriction. Objective. To identify fetal growth disorders and their association with maternal glycemic control and laboratory markers of renal function. Methods. Sixty pregnant women with DM1 were prospectively followed from the first trimester in an observational cohort. The association between birthweight classification with the following parameter were investigated: glycemic mean, frequency of hypo and hyperglycemia, frequency of severe hyper and hypoglycemia, glycated hemoglobin, fructosamine, uric acid, creatinine and proteinuria of 24 hours. The prediction of fetal growth was also investigated. Results. Abnormal fetal growth was observed in 41% (n= 25). Large for gestational age (LGA) was observed in 31.7% (n=19) and small for gestational age (SGA) in 10% (n= 6). High values of glycemic mean (p = 0.006), low frequency of hypoglycemia (p = 0.027) and high frequency of hyperglycemia (p = 0.014) were significantly associated with LGA fetal growth in the third trimester. In all trimesters, the SGA fetal growth was significantly associated with higher serum values of uric acid, creatinine and proteinuria of 24 hours. The prediction model for fetal growth, using values of glycemic mean and creatinine, was significant in the third trimester with an accuracy of 80.3%. Conclusions. The maternal glycemic control and the laboratory markers of renal function associated with the fetal growth disorders in pregnancies with DM1 were identified. Using these parameters it was possible to predict with a good accuracy the fetal growth in DM1

Introdução: A estimulação elétrica nervosa transcutânea (TENS) é uma modalidade terapêutica analgésica, não invasiva e que não apresenta interação com medicamentos. É definida como qualquer dispositivo de estimulação que emita correntes elétricas através da superfície intacta da pele. É utilizada por fisioterapeutas para o manejo sintomático de dor aguda ou crônica em locais de atendimento à saúde. Objetivo: Avaliar os efeitos da estimulação elétrica nervosa transcutânea de baixa e alta frequência sobre o abdome gravídico da camundonga Swiss durante toda a prenhez. Métodos: Trinta camundongas prenhas, pesando 20 a 36 gramas, da linhagem Swiss, foram divididas ao acaso em três grupos (n=10): grupo placebo (P), onde os animais foram submetidos à simulação da eletroestimulação com o aparelho devidamente desligado, diariamente por 20 minutos, a partir do dia zero até o 20° dia da prenhez; grupo experimental baixa frequência (BF), onde os animais foram submetidos ao tratamento por eletroestimulação diariamente por 20 minutos, a partir do dia zero até o 20° dia da prenhez com o aparelho previamente programado com os seguintes parâmetros: frequência baixa de 10 Hertz (Hz), duração de pulso de 200 microssegundos (?s) e a intensidade sensorial foi considerada a partir de 2.0 miliamperes (mA); grupo experimental alta frequência (AF), no qual os animais foram submetidos ao tratamento da eletroestimulação diariamente por 20 minutos, a partir do dia zero até o 20° dia da prenhez com o aparelho previamente programado com os seguintes parâmetros: frequência alta de 150 Hz, duração de pulso de 200 ?s e a intensidade sensorial foi considerada a partir de 2.0 mA. As camundongas foram pesadas no dia zero, 7°, 14° e 20° dia para verificar o ganho ponderal semanal. No 20º dia as camundongas do grupo P, BF e AF foram anestesiados com xilazina e ketamina na dosagem de 0,1 mg/kg e 0,2 mg/kg, respectivamente, e sacrificadas para verificar o número de implantações, reabsorções, de fetos, de placentas e malformações fetais maiores e externas. Resultados: Não houve diferenças estatisticamente significantes ao analisar os ganhos ponderais semanais das matrizes (zero, sétimo, 14° e 20°), número de fetos, placentas, reabsorções, implantações e malformações comparando os dados do grupo placebo e os experimentais. Conclusões: As camundongas submetidas ao tratamento com a TENS de baixa e alta frequência, sobre o abdome gravídico, não apresentaram efeitos deletérios ou teratogênicos / Introduction: The transcutaneous electric nerve stimulation (TENS) is a non-invasive analgesic therapeutic modality, and does not present interaction with drugs. It is defined as any stimulation device that transmits electrical currents through the intact skin surface. It is used by physical therapists for the symptomatic management of acute or chronic pain of benign origin in health care facilities. Aim: To assess the effects of low and high frequencies of Transcutaneous Electric Nerve Stimulation (TENS) on the pregnant abdomen of albine female mice over the pregnancy period. Methods: Thirty pregnant mice, weighting between 20 to 36 grams, of the Swiss lineage were randomly divided in three groups (n=10): Placebo group (P), in which the animals were submitted to a daily electric stimulation treatment with the device unplugged, for 20 minutes, from day zero until the 20th day of pregnancy; Low Frequency Experimental group (LF), in which the animals were submitted to a daily electric stimulation treatment, for 20 minutes, from day zero until the 20th day of pregnancy and the device was previously set up with the following parameters: low frequency 10 Hertz (Hz), 200 microseconds pulse (?s) length and the sensorial intensity will be considered from 2.0 milliamperes (mA) and up, High Frequency Experimental group (HF), in which the animals were submitted to a daily electric stimulation treatment for 20 minutes from day zero until the 20th day of pregnancy and the device was set up with the following parameters: high frequency 150 Hz, 200 ?s length and the sensorial intensity will be considered from 2.0 mA and up. The mice were weighed on day zero, 7th, 14th and 20th days in order to check the weekly pondered weight gain. On the 20th day the mice of the P, LF and HF groups were anesthetized with xylazine and ketamine with a dosage of 0,1 mg/kg and 0,2 mg/kg, respectively. After that, the mice were sacrificed in order to check the number of implantations, reabsorptions, fetus, placenta and major and external fetal malformations. Results: There were no significant statistic differences when analyzing the weekly pondered gain of the matrices (zero, 7th, 14th, 20th), number of fetus, placenta, reabsorptions, implantations and malformations and comparing data of the placebo group to the experimental groups. Conclusions: The mice submitted to the low and high frequency TENS treatment on the pregnant abdomen do not presented neither deleterious nor teratogen effects

O objetivo deste estudo foi verificar o poder de predição da medida da altura uterina para a restrição do crescimento fetal (RCF), por meio da curva de MARTINELLI et al. (2001), tendo como limite o percentil 5 e 10 para a idade gestacional e comparar com a curva de BELIZÁN et al. (1978). Entre julho de 2000 e fevereiro de 2003, 238 gestantes de alto risco foram submetidas a medidas de altura uterina, da 20a à 42a semana de gestação. Todas possuíam idade gestacional confirmada por ultra-sonografia precoce. A confirmação do diagnóstico de RCF foi dada após o nascimento pela curva de RAMOS (1983). Entre as gestantes, 50 (21,0%) tiveram recém-nascidos pequenos para a idade gestacional. O mesmo observador realizou 1617 medidas de altura uterina, com fita métrica, da borda superior da sínfise púbica ao fundo uterino. Para a ocorrência de RCF, considerando um exame positivo se uma medida de altura uterina encontrava-se abaixo do percentil 10 para a idade gestacional na curva de MARTINELLI et al. (2001), a sensibilidade (S) foi de 78%, especificidade (E) de 77,1%, valor preditivo positivo (VPP) de 47,6% e valor preditivo negativo (VPN) de 92,9%. Utilizando como limite o percentil 5, foram obtidos S= 64%, E= 89,9%, VPP= 62,7% e VPN= 90,4%, para o diagnóstico da RCF. Utilizando-se a curva de BELIZÁN et al. (1978) e considerando positivo exame com um valor abaixo do percentil 10 para a idade gestacional, os resultados encontrados foram S= 54%, E= 97,3%, VPP= 84,4% e VPN= 88,8% para a identificação da RCF. Comparada à curva de BELIZÁN et al. (1978), a curva de altura uterina de MARTINELLI et al. (2001) apresentou maior sensibilidade e valor preditivo negativo, consistindo em método de triagem mais adequado para a RCF / The aim of this study was to correlate uterine height measurements below the 5th and 10th percentiles using MARTINELLI et al. (2001) curve to fetal growth restriction (FGR) and to compare with the BELIZÁN et al. (1978) curve. During the period of July 2000 and February 2003, 238 pregnant women of high risk were submitted to uterine height measurements between the 20th and 42nd weeks of gestation. The whole group had well-known gestational age, confirmed by early ultrasound. The diagnosis of FGR was confirmed after birth according to RAMOS (1983). Among these women, 50 (21,0%) gave birth to light for gestational age infants. The same observer, using tape measure, performed 1617 uterine height measurements, from the upper border of the symphysis pubis to the fundus uteri. For the diagnosis of FGR, being considered as positive the exam with measurements below the 10th percentile according to MARTINELLI et al. (2001) curve, the sensitivity (SE), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) were 78,0%, 77,1%, 47,6% and 92,9%, respectively. For the 5th percentile, this curve showed SE= 64,0%, SP= 89,9%, PPV= 62,7% and NPV= 90,4% for the detection of FGR. The BELIZÁN et al. (1978) curve, having the 10th percentile as the limit, yielded SE= 54,0%, SP= 97,3%, PPV= 84,4% and NPV= 88,8% for the identification of FGR. We conclude that, when used for screening FGR, the MARTINELLI et al. (2001) curve showed greater sensitivity and negative predictive value, and presents better results than that of Belizán et al. (1978)

INTRODUÇÂO: O funcionamento da tireóide fetal se inicia em torno de dez semanas de vida embrionária, e está intimamente relacionado ao funcionamento tireoidiano materno. Em gestantes com doenças tireodianas (principalmente hipertireoidismo), a passagem de anticorpos e medicações maternas pode provocar o mau funcionamento da tireóide fetal, acarretando bócio fetal. Além disso, algumas doenças fetais podem cursar com bócio antenatal. O funcionamento inadequado da tireóide fetal pode ter conseqüências severas (restrição de crescimento intra-uterino, craniosinostose, alterações na produção de líquido intra-âmniótico, insuficiência cardíaca ou até óbito fetal). Além disso, o bócio fetal avançado pode funcionar como obstrução à via de parto, podendo acarretar problemas na evolução do parto. A ultra-sonografia da tireóide fetal vem sendo descrita como um bom método para avaliação de tireóide fetal, porém existem poucas curvas de normalidade da tireóide fetal descritas atualmente, nenhuma em população brasileira. O objetivo desse estudo é construir curvas de normalidade do perímetro, área e diâmetro transverso da tireóide fetal em população brasileira através da utilização da ultra-sonografia bidimensional. MÉTODOS: Foram avaliadas 239 gestantes sem doenças sistêmicas e sem história de doença tireoidiana do pré-natal do Hospital das Clínicas da Universidade de São Paulo. Todas as gestantes realizaram dosagem de TSH durante a gestação para descartar doença tireoidiana. A idade gestacional foi calculada pela data da última menstruação, e confirmada por ultrasonografia de primeiro ou segundo trimestre. Foram construídas curvas de normalidade do perímetro, área e diâmetro transverso da tireóide fetal. Das 239 pacientes inicialmente avaliadas, 43 (18%) foram excluídas. A prevalência de hipotireoidismo subclínico entre as gestantes foi de 0,9%, e a de hipotireoidismo franco de 2,2%. Em 5,4% das pacientes não foi possível a visualização adequada da tireóide fetal. Foram incluídas 196 pacientes. A avaliação foi realizada por dois operadores independentes. Foram realizadas três medidas de cada parâmetro, e considerada a média dos valores para a construção das curvas. Para o cálculo da variação intra-observador, foram avaliadas 159 pacientes e realizadas três medidas subseqüentes de cada parâmetro. Para o cálculo da variação inter-observador foram avaliadas 34 pacientes, nas quais cada operador realizou uma medida de cada parâmetro. RESULTADOS: Foram construídas curvas de normalidade do perímetro (P), área (A) e diâmetro transverso (DT) da tireóide fetal em relação à idade gestacional (IG) em nossa população de gestantes. As equações que melhor representaram a média esperada por idade gestacional foram equações lineares: P = 0,146 x IG; A = -1,289 + 0,085 x IG; DT = 0,054 x IG. / INTRODUCTION: The functioning of fetal thyroid initiates around ten weeks of embryonic life, and is intimately related to maternal thyroid functioning. In pregnant women with thyroid disease (especially hyperthyroidism), maternal antibodies and medications provoke malfunctioning of the fetal thyroid gland, causing fetal goiter. Moreover, primary fetal anomalies may course with antenatal goiter (i.e.: congenital fetal hypothyroidism). The inadequate functioning of fetal thyroid causes severe consequences such as intrauterine growth restriction, craniosinostosis, altered production of intra-amniotic liquid, cardiac insufficiency and even fetal death. Moreover, large fetal goiters may cause a mass effect, causing difficulties during vaginal delivery. Ultrasound evaluation of fetal thyroid has recently been described as a sensible method for fetal screening of thyroid anomalies. Few normality curves have been described until now, none of them in Brazilian fetuses. The aim of this study was to build normality curves of fetal thyroid perimeter, area and transverse diameter through the use of bidimensional ultrasonography. METHODS: 239 pregnant women without systemic disease and without previous thyroid disease were evaluated in the prenatal care unity of the Hospital of the Clinics of the University of São Paulo. All women had TSH measured during pregnancy in order to exclude thyroid disease. Gestational age was calculated by the date of the last menses, and confirmed by first or second trimester ultrasonography. Normality curves of fetal thyroid perimeter, area and transverse diameter were constructed. From the 239 patients initially evaluated, 43 (18%) were excluded. The prevalence of thyroid hormone disorders was 0.9% for subclinical hypothyroidism, and 2.2% for hypothyroidism. One patient had a TSH value below normal. In 5.4% of patients the adequate visualization of fetal thyroid was not possible. The evaluation was carried out in 196 patients by two independent operators. Three different measures of each parameter were performed. The average values were used to build the curves. Intra-observer variation analysis was made using 159 patients in whom three measures were made for each parameter. For the inter-observer variation, 34 patients were evaluated. RESULTS: Normality curves of the perimeter (P), area (A) and transverse diameter (TD) were constructed in relation to gestational age (GA) in our population. The 95% confidence interval was calculated. The equations that better represented the average value expected for each gestational age were linear regressions: P = 0,146 x GA; A= -1,289 + 0,085 x GA; TD = 0,054 x GA. The method was considered to be reproductive.