Dissertation for MMed (Obstetrics and Gynaecology) and FCOG Part II / Hypertensive disorders in pregnancy are common and their incidence appears to be on the increase. Preeclampsia is a multi-organ, heterogeneous disorder of pregnancy associated with significant maternal, fetal and neonatal morbidity and mortality. Because preeclampsia is a progressive disorder, invariably delivery remote from term is often necessary to halt disease progression to benefit the mother and fetus. Objectives:  To determine the maternal outcomes in women with severe preeclampsia  To determine fetal and neonatal outcomes of infants born preterm Methods This was a prospective, descriptive study performed in three academic hospitals affiliated to the University of the Witwatersrand in Johannesburg. Data was collected from women with severe preeclampsia, who delivered between gestational ages of 26 weeks and 33 weeks, with a minimum neonatal weight of 500g as determined by sonography. Results: In the sample of 92 patients enrolled, there were two maternal deaths as a result of severe preeclampsia. Eclampsia and HELLP syndrome were the most frequently observed maternal complications at 34% and 49% respectively. Caesarean section was the most frequent method used to expedite delivery in 84% of women. Of the 97 babies delivered, 20% were confirmed intra-uterine fetal deaths, 7% demised during the early neonatal period and a there was a 40% very low birth weight rate. Conclusion: Despite interventions to reduce maternal and neonatal morbidity and mortality in our setting, our outcomes are similar to those observed in other parts of the world.

Alcohol use during pregnancy is common and its consequences often result in a broad range of negative, lifelong developmental outcomes. This study describes the effects of prenatal alcohol exposure and interacting socio-demographic factors on early childhood development. One hundred and twenty one children from the Northern Cape, South Africa, were clinically examined using standard diagnostic procedures and assessed using the Griffiths Mental Development Scales (GMDS/ER) at 7-12 months (Time 1) and 5 years of age (Time 2). Participants were assigned to either: a Fetal Alcohol Syndrome (FAS/Partial Fetal Alcohol Syndrome (PFAS); a Prenatal Alcohol Exposed (PAE); or a Control group based on the diagnosis at 5 years. Mothers/caregivers were interviewed to ascertain socio-demographic information, including prenatal alcohol exposure. During infancy, the FAS/PFAS group showed significantly lower gross motor and language abilities, with delays in higher-order executive functioning becoming more apparent with age. No significant differences were noted during infancy between the PAE and Control groups over any developmental subscales. However, with age, higher-order executive function delays were reported in the PAE group. Performance on the infant and child versions of the GMDS was not significantly correlated, suggesting that the tests may be measuring different developmental constructs. Lower maternal education, unemployment and later recognition of pregnancy were associated with reduced social adaptive functioning, and language and eye hand coordination abilities, irrespective of amount of prenatal alcohol exposure over both time points. Larger anthropometric birth measurements and longer duration of breastfeeding were significantly related to increased performance on the GMDS at 5 years within the groups exposed to prenatal alcohol. Socio–demographic variables are likely to complicate developmental profiles for all three groups, with prenatal and postnatal nutrition emerging as possible protective factors for positive developmental outcomes at 5 years of age.

A gemelaridade apresenta algumas intercorrências específicas, como a discordância de peso entre fetos e recém-nascidos (RNs). O objetivo do presente estudo foi predizer a discordância de peso do exame ultra-sonográfico comparada à do parto e avaliar a morbidade e a mortalidade neonatais nas gestações gemelares discordantes quanto ao peso. Este foi um estudo retrospectivo, com levantamento dos casos do período de 1998 a 2004, no Setor de Gestações Múltiplas da Clínica Obstétrica do HCFMUSP. Na avaliação da predição ultra-sonográfica, foram inseridas 221 gestações gemelares e, na avaliação da morbidade e da mortalidade, 151 gestações com partos nessa instituição. A discordância de peso foi definida como >= 20%, sendo excluídos os casos de malformações fetais (n=43) e da Síndrome da transfusão feto-fetal (n=24). Para análise da adequação do peso ao nascimento, utilizou-se a curva de Alexander et al., 1998, para gêmeos. No estudo da predição, foram utilizados quatro intervalos de tempo em relação ao parto (0 a 7 dias - n = 96; 8 a 14 dias - n = 66; 15 a 21 dias - n = 58; 22 a 28 dias - n = 59 gestações), somando 279 avaliações. No grupo de 0 a 7 dias, a estimativa da sensibilidade foi de 93,6%, especificidade de 79,4%, valor preditivo positivo de 89,2%, valor preditivo negativo de 87,1% e acurácia de 88,6%. Nos demais grupos, a sensibilidade e a acurácia foram de 95,8% e 84,9%, 95,6% e 84,5%, 90,9% e 84,8%, respectivamente. Em relação à morbidade, 111 gestações eram concordantes (73,5%) e 40 discordantes quanto ao peso. No grupo discordante, 75% das gestações gemelares apresentaram pelo menos um recém-nascido com Restrição de Crescimento Fetal (RCF). Nesta análise, as gestações gemelares concordantes monocoriônicas obtiveram menor média de idade gestacional no parto (34,3 versus 36,2 semanas, p=0,004), menor peso médio (2067 versus 2334 gramas, p=0,0016) e maior tempo de internação (10,6 versus 7,3 dias, p=0,0023) que as gestações concordantes dicoriônicas. Nas gestações discordantes, não houve diferença significativa em relação à corionicidade. As gestações discordantes, com pelo menos um RN abaixo do percentil 10, apresentaram menor média de idade gestacional (35,2 versus 36,8 semanas, p=0,009) e maior tempo de internação (17,5 versus 8,2 dias, p=0,026). Não foi observada diferença significativa de morbidade e mortalidade entre RNs concordantes e discordantes, com pesos entre os percentis 10 e 90. Os fetos menores das gestações discordantes demonstraram maior freqüência de índice de Apgar inferior a 7 (27,5% versus 7,5%, p=0,01). A avaliação da mortalidade não demonstrou diferença significativa em relação aos grupos concordantes (3,7%) e discordantes (4,5%; p = 1,00). No presente estudo, conclui-se que os quatro grupos apresentaram adequada correlação entre a discordância de peso à ultra-sonografia e no nascimento, porém com melhor predição até sete dias antes do parto. A morbidade neonatal esteve relacionada à RCF do menor feto. A discordância de peso e a corionicidade não interferiram na mortalidade neonatal. / The aim of this study was to evaluate the ability of prenatal ultrasound scans to predict fetal growth discordance in twin pregnancies and perinatal morbidity/mortality associated with these cases. This was a retrospective study (1998-2004) involving twin pregnancies that were scanned and had their delivery at our Institution (HCFMUSP). Cases with fetal malformations (n=43) or twin to twin transfusion syndrome (n=24) were excluded. The study of ultrasound scans consisted of 221 twin pregnancies. The final morbidity/mortality study group consisted of 151 twin pregnancies. Birth weight was evaluated based on twin growth charts published by Alexander et al (1998) and weight discordance as a difference >= 20%. Small for gestacional age (SGA) was defined as birth weight below the 10th centile. The study of ultrasonographic prediction of interwin discordance was made using four different intervals between ultrasound examination and delivery (0 to 7 days, n = 96; 8 to 14 days, n = 66; 15 to 21 days, n = 58; 22 to 28 days, n = 59 pregnancies), with a total of 279 ultrasound examinations. In group 0 to 7 days, the sensitivity was 93,6%, specificity was 79,4%, positive predicted values was 89,2%, negative predicted values was 87,1% and accuracy was 88,6%. In the groups 8 to 14 days, 15 to 21 days and 22 to 28 days the sensitivity and accuracy were 95,8% and 84,9%, 95,6% and 84,5%, 90,9% and 84,8%, respectively. Birthweight discordance was observed in 40 sets of twins (26.5%) and 12 cases were monochorionic MC (30%). Twenty five cases (22.5%) in the non discordant group were MC. In the non discordant group, monochorionic pregnancies showed lower gestational age at delivery (34.3 versus 36.2 wks, p=0.004), lower mean birth weight (2067g versus 2334g, p=0.0016) and longer length of stay in hospital (10.6 versus 7.3 days, p=0.0023) compared to dichorionic twins. In the group with twin birthweight discordance, there were no significant differences between MC and DC pregnancies and 75% of the cases had at least one newborn with SGA. These cases were showed lower gestational age at delivery (35.2 versus 36.8wks, p=0.009) and longer length of stay in hospital (17.5 versus 8.2 days, p=0.026). In the discordant group, the smaller twin had a higher frequency of first minute Apgar score < 7 (27.5% versus 7.5%, p=0.01). Perinatal mortality rate was similar in both groups (discordant 4.5% and concordant 3.7%, p=1.0). There were no significant differences in morbidity and mortality between concordant and discordant twins when birth weight was between the 10 th and 90 th centile. In conclusion, there was a good correlation between fetal growth discordance predicted by prenatal scan and actual birth weight discordance. Neonatal morbidity was related to SGA. Excluding fetal malformation and TTTS cases, birth weight discordance in twin pregnancies is not a significantly associated with neonatal mortality.

INTRODUÇÃO: Gastrosquise é uma malformação da parede abdominal do feto e uma das principais complicações relacionadas à restrição de crescimento fetal. Objetivo principal: avaliar o padrão de crescimento de fetos com gastrosquise para cada parâmetro biométrico. Objetivo secundário: avaliar o déficit de crescimento em três períodos gestacionais e predizer recém-nascidos pequenos para idade gestacional a partir de medidas de parâmetros biométricos abaixo do percentil 10. MÉTODOS: Este é um estudo do tipo coorte retrospectivo. Foram selecionados 70 casos para avaliação do padrão de crescimento. As medidas de cada parâmetro biométrico: circunferência cefálica, circunferência abdominal, comprimento femoral, razão circunferência cefálica/circunferência abdominal e peso fetal estimado foram plotadas em um gráfico de dispersão para comparação com a curva de referência. A diferença porcentual entre as médias das medidas dos fetos com gastrosquise em relação aos normais foi determinada. Para a avaliação do déficit de crescimento foram incluídos 59 casos, com ao menos um exame em cada período gestacional (I:20 a 25 semanas e 6 dias; II:26 a 31 semanas e 6 dias; III: 32 semanas até o parto). O déficit de cada parâmetro biométrico foi obtido a partir da comparação entre os períodos gestacionais. Para a predição de recém-nascido pequeno para idade gestacional foram utilizadas as medidas abaixo do percentil 10 de cada parâmetro biométrico nos períodos I e II. RESULTADOS: Na avaliação do padrão de crescimento, observa-se diferença significativa entre os fetos com gastrosquise e fetos normais a partir de 20 semanas de gestação (p<0,005). Na avaliação do déficit de crescimento, apenas peso fetal estimado apresentou diferença significativa (p=0,030). O porcentual de fetos com peso fetal estimado abaixo do percentil 10 no período 2 foi 40% maior do que no período 1, e 93% maior no período 3 do que no 1. Na predição de recémnascidos pequeno para idade gestacional, apenas a circunferência cefálica (razão chance= 6.07; sensibilidade= 70.8%; especificidade= 71.4%) e a circunferência abdominal (razão chance=0,558; sensibilidade= 41,7%; especificidade= 80%) no período II, foram consideradas. CONCLUSÃO: Fetos com gastrosquise apresentam medidas dos parâmetros biométricos significativamente menores do que as de fetos normais, a partir de 20 semanas de gestação. Na avaliação do déficit de crescimento, observa-se maior incidência de restrição de crescimento fetal nos períodos II e III em comparação ao período I. É possível predizer recém-nascidos com baixo peso ao nascimento, a partir de medidas de circunferência cefálica e circunferência abdominal, abaixo do percentil 10 no período II / INTRODUCTION: Gastroschisis is a congenital abdominal wall defect of the fetus and one of its main complications is related to fetal growth restriction. OBJECTIVES: Primary: To evaluate the growth pattern of fetuses with gastroschisis according to each biometric parameter; Secondary: to evaluate growth deficit in three gestational periods and to predict low birth weight from measures of biometric parameters below the 10th percentile. METHODS: This is a retrospective cohort study. We selected 70 cases for evaluation of the growth pattern. The measurements of each biometric parameter: head circumference, abdominal circumference, femur length, head circumference/abdominal circumference ratio and estimated fetal weight were plotted in a growth chart for comparison with the curve of normality. The percentage difference between the mean values of the fetuses with gastroschisis in relation to normal fetuses was then determined. For the evaluation of growth deficit 59 cases with at least one exam in each gestational period (I: 20 to 25 weeks and 6 days; II: 26 to 31 weeks and 6 days; III: 32 weeks until delivery) were included. The deficit of each biometric parameter was obtained from the comparison between these gestational periods. For the prediction of low birth weight, the measures below the 10th percentile of each biometric parameter in periods I and II were tested. RESULTS: In the evaluation of the growth pattern a significant difference between the fetuses with gastroschisis and normal fetuses from 20 weeks of gestation (p < 0.005) is observed. In the evaluation of growth deficit only estimated fetal weight showed a significant difference (p= 0.030). The percentage of fetuses with estimated fetal weight values below 10 percentile in period 2 was 40% higher than that in period I, and 93% higher in period III than in I. In the prediction of low birth weight, only head circunference (odds ratio= 6.07; sensitivity= 70.8 %; specificity = 71.4 %) and abdominal circunference (odds ratio= 0.558; sensitivity = 41.7 %; specificity = 80 %) in period II were predictive. CONCLUSION: Fetuses with gastroschisis show biometric parameters measures significantly smaller than the measures of normal fetuses with 20 weeks of gestation and/or more. In the evaluation of growth deficit, there is a higher incidence of fetal growth restriction in periods II and III. It is possible to predict newborns with low birth weight from measures of head circunfernce and abdominal circunference below the 10th percentile in period II

Submitted by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-06-30T17:45:25Z No. of bitstreams: 1 DIS_CAROLINE_MACHADO_DARTORA_COMPLETO.pdf: 2382956 bytes, checksum: 7909b8673eceb86e85c844ad70245e3b (MD5) / Made available in DSpace on 2017-06-30T17:45:25Z (GMT). No. of bitstreams: 1 DIS_CAROLINE_MACHADO_DARTORA_COMPLETO.pdf: 2382956 bytes, checksum: 7909b8673eceb86e85c844ad70245e3b (MD5) Previous issue date: 2017-03-29 / The statistical nature of the physical process involved in Nuclear Medicine exams makes the use of Monte Carlo methods an useful tool for deposited energy and absorbed dose calculations on organs, mainly for risk-benefits assessment. Pregnant are important target for risk-benefit assessment, due the fetus exposure to radiation. Nuclear Medicine exams may generate fetus dose, either by irradiation due to maternal organs, or by activity that crosses the placenta and accumulates in fetus. Usually, both internal dosimetry estimation software and virtual anthropomorphic simulators are proprietary. The aim of this research is to investigate the use of GATE (Geant4 Application for Emission Tomography) application of Monte Carlo method on internal dosimetry calculation in simulated Nuclear Medicine image exams in pregnant woman. First, an evaluation of the use of GATE was performed to build dose maps with simple geometries. An evaluation of the radiation interaction behavior with respect to available GATE source types of configuration of single photon (99mTc) and positrons (18F) emitters was also performed. Dose estimation on a fetus was performed by simulation of a [18F] FDG distribution in a virtual 24-week pregnant woman simulator, Katja, with data based on literature. It was evaluated the impact on fetal dose of the mother bladder uptake of , placenta uptake, maternal organs irradiation and dose generated due to the uptake only in the fetus. The total estimated dose for a fetus on 24 weeks was 0.0122 mGy/MBq, in agreement with published data. Several individual dose contributions in the fetus that are not commonly found in the literature, such as the contribution due to the bladder (13%) and placenta (0.53%) were obtained. The contribution in total fetal dose of the activity only in the organs of the fetus was analyzed, resulting in 3.8%, where 56% is due only to positron emitted by the source. In conclusion, GATE generates dose maps that can be used as a method of dose estimation in pregnant women in MN scans, giving detailed information about the individual contributions of maternal organs uptake. / A natureza estat?stica dos processos f?sicos nos exames de Medicina Nuclear faz com que o uso de m?todos de Monte Carlo seja ?til para c?lculos da energia depositada e da dose absorvida nos ?rg?os, principalmente para avalia??o de risco-benef?cio. Gr?vidas s?o um alvo importante para avalia??o risco-benef?cio devido ? exposi??o do feto ? radia??o. Exames de Medicina Nuclear podem gerar dose no feto, tanto pela irradia??o devido ? atividade nos ?rg?os maternos, como pela atividade que atravessa a placenta e se acumula no feto. Usualmente, os softwares de estimativa de dosimetria interna e os simuladores antropom?rficos virtuais s?o propriet?rios. O objetivo desta pesquisa ? investigar o uso do aplicativo gratuito de simula??o de Monte Carlo denominado GATE (Geant4 Application for Emission Tomography), no c?lculo da dosimetria interna em exames simulados de Medicina Nuclear em mulheres gr?vidas. Inicialmente, foi realizada uma avalia??o do uso do GATE na constru??o de mapas de dose com geometrias simples e o comportamento para diferentes configura??es dos tipos de fontes no aplicativo para emissores de f?ton ?nico (99mTc) e p?sitrons (18F). A estimativa de dose no feto foi realizada atrav?s da simula??o de uma distribui??o de [18F]FDG em um simulador virtual gratuito de mulher gr?vida de 24 semanas, denominada Katja, com distribui??o de atividade baseada na literatura. Foi investigado o impacto, na dose fetal total, da capta??o e esvaziamento da bexiga da m?e, da placenta, da irradia??o do feto pelos ?rg?os maternos e dose gerada devido ? atividade captada somente pelo feto. A dose estimada total para um feto de 24 semanas foi de 0,0122 mGy/MBq, estando de acordo com a m?dia dos trabalhos publicados. Obtiveram-se contribui??es individuais ? dose no feto que n?o s?o comumente encontradas na literatura, tais como a dose devido ? bexiga (13%) e ? placenta (0,53%). Foi analisada a contribui??o da atividade captada somente nos ?rg?os do feto na dose total no feto, que resultou em 3,8%, sendo que 56% correspondem ? dose devido aos p?sitrons emitidos pela fonte. Conclui-se que o aplicativo GATE gera mapas de dose que podem ser utilizados como um m?todo de estimativa de dose de mulheres gr?vidas em imagens de Medicina Nuclear, fornecendo informa??es detalhadas das contribui??es individuais da capta??o nos ?rg?os da m?e ? dose.

A Cassia Angustifolia (Sene) é considerada um planta medicinal indicada principalmente para fins purgativos e constipação intestinal, no entanto, destaca-se também pelo seu uso como abortiva e emenogoga. Plantas medicinais tardicionalmente usadas para induzir o aborto são usadas em altas doses, podendo causar efeitos tóxicos e prejudiciais ao feto e ao corpo da mulher. O principal constituinte químico da CAssia angustifolia é a antraquinonas, onde o metabólico secundário chamado de diantrona (Senosídeos A a F) e também outros compostos como glicosideos, naftalênicos e flavonoides. Estudos recentes destacam que esses compostos antraquinônicos aumentam o influxo de sangue para o útero e seus anexos, provocam intoxicação placentária, aumentando assim o risco de perda fetalocasionando principalmente por disfunção do fosforilação oxidativa mitocondrial. Na interface - materno fetal existem células trofoblásticas gigantes (CTGs) que são de grande relevância que são de grande relevância para a manutenção do equilíbrio existente entre o feto e a gestante. Elas . Elas. Elas se localizam entre os tecidos maternos e fetais durante todo o período se localizam entre os tecidos maternos e fetais durante todo o período. Elas se localizam entre os tecidos maternos e fetais durante todo o período gestacional, adquirindo funções de invasidade, hematotrófico e fagocitário. Existem também zx células natural Killer uterinas (uNK) cuja principal função é a liberação de citocinas com IFN-Y que leva à dilatação ds artérias espiraladas deciduais e manutennção de integridade decidual, contribuindo, portanto, para o desenvolvimento normal da decídua e da placenta. Neste contexto, o presente trabalho buscou avaliar os possiveis efeitos da administração do extrto hidroalcoolico de Cassi angustifolia (Sene) em diferentes períodos da gestação de camundongos, tanto no organismo materno como cno concepto, bem como os efeitos comportamentaios eos efeitos nas células uNK e CTGs desses animais por meio dos estudo morfológico, citoquímico e estereológicos. Realizou-se também o perfil cromatográfico a fim de averiguar os principais constituites quimicos do extrato. O tratamento com extrato provocou alterações morfologicas bastante evidentes, de tal forma que foi possível identificar a presença de focos hemorrágicos e reabsorções uterinas. Verificamos que o CTGs se localizavam em meio ao infiltardo sanguineo, ou até mesmo ao redor de um inicio de necrose, exercendo assim um papel fagocitário frente à aitividade inflamatória e remodelação tecidual. As células NKu apresentaram um aumento significativo a região endometrial mais distante do embrião. Esses achados sugerem que existe uma cinética no proecesso de perda e reabsorção embrionária nos naimais tratados durante o prenhez que se inicia com o processo hemorrágico, morte e reabsorção embronária e que os senosídeos A e pode estar associado a estes efeitos. Portanto o uso de Cassia angustifolia durante os periodos de desenvolvimento embrionário apresenta um grande risco para a mãe e para o feto / Cassia angustifolia (Senne), is usually associated with its effects on the gastrointestinal tract where besides being used as purgative. The popular culture associated this plant with abortion and emmenagogue effects. Mostplants used as emmenagogu eare the same used as abortion, there is no clear distinction between the two intentions. Plants traditionally used to induce abortion are administered in excessively high doses, which can cause toxic effects to the woman's body. The main chemical constituents of Cassia angustifolia are diantronas-8-8'-called diglicosídeos senossides AF, with a predominance of A senossides. This components can inducing uterine contractions, increase blood flow to the uterus and its appendages, increasing the risk offetal loss, disruption of the mitochondrial oxidative phosphorylation. Trophoblast giant cells (TGC) present in maternal interface - fetal, are of great relevance to the balance between the fetus and mother. There are also uterine Natural Killer cells (UNK) whose main function is the release of cytokines such as IFN-? which leads to dilation of the decidual spiral arteries and maintenance of decidual integrity, contributing thus to the normal development of the decidua and placental. In this context, the present study evaluated the possible effects of administration of the hydroalcoholic extract of Cassia angustifolia (Senna) in different periods of pregnancy in mice, both in the mother and the fetus, as well as behavioral effects and the effects on cells and TGCs and UNK by morphological, cytochemical and stereological studying. There was also thechromatographic profilein order to as certain the chemical constituents of the extract. We found that theTGCs are locatedin the midst of blood infiltration, or even around an early necrosis, thereby exerting a role phagocytic activity against the inflammatory and tissue remodeling. UNK cells showed a significant increase in endometrial farthest region of the embryo. Furthermore, the treatment showed behavioral changes in females treated with the corresponding extract.The morphological and histological analysis showeda important prejudicial effect of senne extract in mouse pregnancy and after phytochemical analysis we can supposed that this effects was caused probably by the A and B healthy senossídeos (anthraquinone derivatives) the main constituents of Senne extract. The results support an important risk of the indiscriminate use of Senneforpregnant woman / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

Durante a gestação, observamos um influxo de linfócitos para a região mesometrial de camundongos denominados de células Natural Killer Uterinas (uNK). Estas células desempenham funções importantes no início da gestação, como a manutenção da tolerância materna ao feto, angiogênese e remodelação das artérias espiraladas. Sua capacidade citolítica é reduzida podendo ser ativada caso haja algum desequilíbrio entre citocinas pró e anti-inflamatórias durante a gestação. A ciclofosfamida é um medicamento usado no tratamento de doenças autoimunes e câncer. Trata-se de um teratógeno, sendo que seu uso pode levar a alterações macroscópicas e microscópicas, bem como, a redução da viabilidade gestacional em camundongos. O objetivo deste trabalho foi avaliar os efeitos da ciclofosfamida no útero e nas células uNK de camundongos no 10° ddg, por meio da análise morfológica, citoquímica e estereológica concomitantemente a análise da viabilidade gestacional e no comportamento destes animais nos parâmetros de depressão e ansiedade. Nossos resultados mostraram que a ciclofosfamida provoca rapidamente o aumento de células uNK no útero, bem como, diminuição na expressão de N-acetil-D-galactosamina em seus grânulos e superfície, atingindo primeiramente células uNK menos diferenciadas próximas ao mesométrio, e posteriormente células mais diferenciadas em regiões mais próximas ao embrião. Seu efeito drástico na gestação provocou ainda diminuição no número de células uNK após 48h de sua administração, concomitante ao aparecimento de muitos sítios em reabsorção, diminuindo a viabilidade gestacional, bem como, estabelecimento de comportamento semelhante ao doentio nesses animais, causando 100% de perda embrionária diagnosticada em nosso estudo da prenhez a termo / During pregnancy, we observed an influx of lymphocytes into the region of mice mesometrial decidua. These cells are natural killer cells (NK), presenting different behavior and phenotype of circulating NK cells. Uterine Natural Killer Cells (uNK) play important roles in early pregnancy, such as maintaining maternal tolerance to the fetus, angiogenesis and remodeling of the spiral arteries. Their cytolytic capacity is reduced during pregnancy and can be activated if there is an imbalance between pro and anti-inflammatory cytokines. Cyclophosphamide is a drug used in treating autoimmune diseases and cancer. It is known that it is a teratogen, and its use can lead to macroscopic and microscopic changes, as well as the reduction of viability pregnancy in mice. The objective of this study was to evaluate the effects of cyclophosphamide in the uterus and in cells uNK (morphological analysis, cytochemistry and stereological) on the 10th ddg, as well as its effects on behavior related to depression and anxiety. Our results showed that cyclophosphamide rapidly increased uNK cells number in the uterus as well as decreased expression of N-acetyl-D-galactosamine in their granules and surface, early reaching less differentiated uNK cells near the mesometrium, and latter more differentiated uNK cells closer to the embryo. His dramatic effect on pregnancy still caused a decreasing in the number of uNK cells 48 after the drug administration, concomitant with the appearance of many resorption implantation sites, decreasing of gestational viability, as well as, establishment of sickness-like behavior, causing 100% of embryo loss diagnosed in the pregnancy to term. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

by Lam, Sau Hing. / Thesis (M.Phil.)--Chinese University of Hong Kong, 1998. / Includes bibliographical references (leaves 141-158). / Abstract also in Chinese. / Content / Acknowledgements / Abbreviation / Project Objectives / Abstract / Chapter Chapter One --- Introduction / Chapter 1.1 --- The definition of homeobox genes / Chapter 1.2 --- Homeobox genes as a transcription factor / Chapter 1.3 --- Homeobox in Drosophila / Chapter 1.3.1 --- The development of Drosophila / Chapter 1.3.2 --- Maternal genes / Chapter 1.3.3 --- Segmentation genes / Chapter 1.3.4 --- Homeobox genes / Chapter 1.4 --- Family of Hox genes in the mammalian system / Chapter 1.5 --- Some possible chemical mechanism in the cascade system of the Hox genes in the vertebrate / Chapter 1.6 --- Hox (Antp-Class homeobox gene) in mammal / Chapter 1.6.1 --- Labial Like homeobox genes / Chapter 1.6.2 --- Proboscipedia Like homeobox genes / Chapter 1.7 --- Divergent homeobox genes / Chapter 1.7.1 --- Paired (prd) Class / Chapter 1.7.2 --- Even-Skipped (Eve) Class / Chapter 1.7.3 --- Distal-less (Dll) Class / Chapter 1.7.4 --- Muscle-Specific Homeobox (Msx) Class / Chapter 1.8 --- Orphan homeobox gene / Chapter 1.8.1 --- The characteristic of Hox 11 sequence in human and mouse / Chapter 1.8.2 --- Novel homeobox genes related to hox 11 gene family / Chapter 1.8.3 --- The mechanism of HOX 11 inducing gene regulation and signal transduction pathways / Chapter 1.8.4 --- HOX 11 in human / Chapter 1.8.5 --- Hox11 in mouse / Chapter 1.8.6 --- Hox11 L1 in mouse / Chapter 1.9 --- Homeobox gene involved in haematopoiesis / Chapter 1.10 --- Some translocations of homeobox genes in blood lineage / Chapter 1.11 --- The development of mouse / Chapter 1.11.1 --- Early organogenesis / Chapter 1.11.2 --- Nervous system development / Chapter 1.11.3 --- Somite development / Chapter 1.11.4 --- Eye development / Chapter 1.11.5 --- Neural crest cell migration / Chapter 1.11.6 --- Branchial arches development / Chapter Chapter Two --- Materials and methods / Chapter 2.1 --- Mouse Embryos / Chapter 2.2 --- RNA extraction / Chapter 2.3 --- Large plasmid preparation / Chapter 2.4 --- The synthesis of cDNAs using Reverse Transcription / Polymerase Chain Reaction (RT-PCR) and ligation into Bluescript® II KS / Chapter 2.4.1 --- The synthesis of RT-PCR products / Chapter 2.4.2 --- The formation of blunt ends of cDNA / Chapter 2.4.3 --- The ligation of cDNA with plasmid vectors / Chapter 2.4.4 --- Transformation / Chapter 2.4.5 --- The miniprep plasmid purification / Chapter 2.5 --- T7 sequencing / Chapter 2.6 --- Double stranded DNA cycle sequencing of plasmid / Chapter 2.6.1 --- Gel electrophoresis / Chapter 2.7 --- Northern blot / Chapter 2.7.1 --- Preparation of Northern blot / Chapter 2.7.2 --- Hybridization of Northern blot / Chapter 2.8 --- DIG labeled probes in whole mount in situ hybridization / Chapter 2.8.1. --- Preparation of linear DNA to generate riboprobes / Chapter 2.8.2 --- Preparation of DIG labeled riboprobe / Chapter 2.8.3. --- Preparation of embryo powder / Chapter 2.8.4 --- Absorption of antibody / Chapter 2.8.5 --- Embryo preparation / Chapter 2.8.6 --- Embryos staining / Chapter 2.9 --- Sections from whole mount in situ hybridization / Chapter 2.10 --- The radiolabeled section in situ hybridization / Chapter 2.10.1 --- The preparation of paraffin wax block and sample sections / Chapter 2.10.2 --- Slide pretreatment / Chapter 2.10.3 --- Preparation of probe / Chapter 2.10.4 --- Hybridization / Chapter 2.10.5 --- Washing / Chapter 2.10.6 --- Emulsification and development / Chapter 2.11 --- DIG-label in situ hybridization of frozen / Chapter 2.12 --- H&E staining / Chapter Chapter Three --- Results and Discussion / Chapter 3.1 --- Synthesis of Tlx3 probes for use in the section and whole mount in situ hybridization / Chapter 3.1.1 --- Synthesis of the Tlx cDNA by RT-PCR method / Chapter 3.1.2 --- The Tlx3 genomic clone for detecting the developmental expression of Tlx3 by Northern Blot / Chapter 3.1.3 --- The characterization of Tlx3 cDNAs and the sonic hedgehog cDNA / Chapter 3.2 --- Section in situ hybridization using different probes / Chapter 3.2.1 --- Section in situ hybridization using the transcribed riboprobes of Pax2 cDNA / Chapter 3.2.2 --- The transcribed riboprobes of Tlx genomic clones were used to hybridize the section in situ hybridization / Chapter 3.2.3 --- The in situ hybridization of frozen sections of mouse embryos using the transcribed riboprobes from Tlx3 cDNA subclone / Chapter 3.3 --- Expression pattern of Tlx3 on whole mount embryos using both cDNA and genomic probes / Chapter 3.3.1 --- The expression of Tlx3 on whole mount in situ hybridization of the mouse embryos using the antisense probes from the Tlx3 genomic clone / Chapter 3.3.2 --- Whole mount in situ hybridization of mouse embryo using the transcribed riboprobes of Tlx3 cDNA / Chapter 3.3.3 --- The expression pattern of sonic hedgehog on embryos at 8.5 to9.5 dpc / Conclusion / Future prospective / Appendix / Reference / Acknowledgement

Imprinted genes are a subset of loci, positioned on autosomes and the X-chromosome, which are expressed monoallelically in a parent-of-origin specific manner. The influence of such genes on the regulation of embryonic growth and postnatal energy homeostasis is well established. The parental conflict hypothesis predicts that, in utero, paternally-expressed genes will promote maternal resource acquisition and thus growth, whereas maternally-expressed genes will oppose this action, restricting resource investment in a single brood in the interests of the lifetime reproductive success of the mother. Grb10 is an imprinted gene which encodes the cytoplasmic adaptor protein Growth factor receptor bound protein 10. In the majority of tissues, Grb10 is expressed from the maternally-derived chromosome. Consistent with conflict theory, transgenic mice inheriting a disrupted Grb10 allele through the maternal line (Grb10Δ2-4m/+) exhibit embryonic overgrowth, although the mechanisms and signalling pathways responsible for this effect are unclear. Grb10Δ2-4m/+ mice also demonstrate enhanced insulin signalling and improved whole body glucose clearance, consistent with the established role of imprinted genes in the regulation of postnatal metabolism. An integrated LacZ gene-trap in the Grb10Δ2-4 allele failed to fully recapitulate endogenous Grb10 expression, notably within the central nervous system. To address this issue, a second transgenic mouse line, Grb10KO, was created. This allele produced strong LacZ reporter expression in the central nervous system, but only when transmitted through the paternal line (Grb10KO+/p), establishing Grb10 as the only known imprinted gene with a reciprocal imprinting profile between the central nervous system and peripheral tissues. Grb10KO+/p mice exhibit a social dominance phenotype, suggesting distinct roles for maternally- and paternally-expressed Grb10, consistent with their respective sites of expression. The current study characterised the Grb10KO allele at the genetic level, and in doing so, revealed a phenotypic difference between Grb10KOm/p and Grb10Δ2-4m/p mice for which a possible explanation was provided. Importantly, with this knowledge, the current study elucidated the genetic and molecular basis for inconsistencies in reporter expression between the two transgenic lines, identifying a novel tissue-specific enhancer element at the locus. In addition to the central nervous system, this enhancer appeared to be active in the mammary epithelium, identifying a novel site of Grb10 expression, which was pregnancy-dependent and specifically from the maternally-inherited chromosome. Characterisation of the functional significance of expression in this tissue revealed that maternally-expressed Grb10 mediates a supply/demand system between lactating mother and suckling pup, acting as a supply promoter and demand suppressor. This role is inconsistent with conflict theory, but suggests the maintenance of the Grb10 imprint in the mammary epithelium might be associated with improved coadaptiveness between mother and offspring. Intriguingly, in utero, Grb10 is both a demand and supply suppressor. When considered together, these findings suggest a wider role for maternally-expressed Grb10 in the homeostatic control of growth and achievement of optimal fitness.

El documento digital no refiere asesor / Publicación a texto completo no autorizada por el autor / Determina la relación entre el Desarrollo fetal y la Hiperémesis Gravídica en gestantes atendidas en el Hospital San Bartolomé durante el Año 2012. El estudio es observacional, analítico-comparativo, retrospectivo de corte transversal. El tamaño de la población en estudio fue de 57 pacientes con el diagnóstico de hiperémesis gravídica (con HG) y 57 pacientes sin el diagnóstico de hiperémesis gravídica (sin HG). Las fuentes de información son las historias clínicas de aquellas pacientes que acudan al Hospital San Bartolomé durante el año 2012. Para el análisis descriptivo se determinaron medidas de tendencia central y dispersión así como para variables cualitativas se determinaron frecuencias y porcentajes. Para el grado de asociación se utilizó la prueba Chi-cuadrado, con un nivel de confianza (IC) del 95%. La edad promedio de las pacientes fue 26.2 años, con el rango de edad más frecuente de 20-34 años (64%), siendo la mayoría Convivientes (71.9%) y de nivel educativo Secundario (70.2%). El 59.6% de las gestantes con HG tuvieron un IMC Normal y el 24.6% tuvieron Sobrepeso, mientras que el 63.2% de las pacientes sin HG tuvieron un IMC Normal y el 26.3% tuvieron Sobrepeso, sin embargo estas frecuencias absolutas son similares (p=0.875). Respecto a las características psicológicas, el 19.3% de las pacientes con Hiperémesis Gravídica y el 8.8% de las pacientes sin Hiperémesis Gravídica tuvieron ansiedad, la depresión se presentó en el 15.8% de las gestantes con HG y en el 5.3% de las gestantes sin HG (p=0.067). Entre las características obstétricas, la hipertensión inducida por el embarazo (p=0.047) se relacionan con la presencia de Hiperémesis gravídica (14% vs. 3.5%). En la ecografía del III Trimestre, los hallazgos como edad gestacional promedio (p=0.012) y el peso fetal promedio (p=0.019) son en promedio mayores con Hiperémesis Gravídica. El 57.9% de las pacientes con Hiperémesis Gravídica tuvieron parto vaginal al igual que el 63.2% de las pacientes sin Hiperémesis Gravídica (p=0.565). Más de la mitad de los recién nacidos eran de sexo Femenino en el grupo con Hiperémesis Gravídica (50.9%) y sin Hiperémesis Gravídica (56.1%) (p=0.573). El peso de los recién nacidos que predominó fue de 2500-3999 gr en el grupo con HG (78.9%) y sin HG (93%). Existe relación entre el Apgar al minuto y la presencia de hiperémesis gravídica (p=0.008). El 56.1% de las pacientes con HG tuvieron una estancia hospitalaria de 1-2 días similar al 45.6% de las pacientes sin HG (p=0.711). No existe relación entre los resultados maternos como Esofagitis (p=0.315) y falla renal (p=0.309) con la presencia de Hiperémesis Gravídica. El bajo peso al nacer tiene cierta relación con la presencia de Hiperémesis Gravídica (p=0.051) (10.5% vs. 1.8%). La mortalidad Perinatal se presentó solo en el 1.8% de los neonatos del grupo con HG. La conclusión es que en la ecografía del III Trimestre, la hiperémesis gravídica se relaciona con la hipertensión inducida por el embarazo (p=0.047) y el apgar al minuto (p=0.008) y cierta relación con el bajo peso al nacer (p=0.051). La edad gestacional promedio (p=0.012) y el peso fetal promedio (p=0.019) es mayor en los pacientes con hiperémesis gravídica. Existe mayor frecuencia de ansiedad y depresión en las pacientes con Hiperémesis Gravídica pero no es estadísticamente significativa, por último las pacientes mayores a 35 años tienen mayor frecuencia con hiperémesis gravídica. / Trabajo académico