Este projeto de pesquisa trata da elaboração de um sistema de reconstrução de imagens obstétricas de fetos, em aparelhos de ultra-som convencionais, para a visualização dessas imagens em três dimensões utilizando a internet como meio de utilização do sistema, com o principal objetivo de proporcionar aos médicos ginecologistas melhor visualização do formato e das estruturas internas, e em especial da face do feto, através do processo de reconstrução tridimensional feito a partir de um conjunto de imagens bidimensionais capturadas em aparelhos convencionais de ultra-som. O uso clínico deste projeto está previsto para o setor de obstetrícia do Hospital das Clínicas de Ribeirão Preto. / This project of research deals with the laboration of a reconstruction system of obstetrics images of embryos in devices of ultrasound will be conventional the visualization of these images in three dimensions using the internet half of uses of the system, with the main objective provides to the medical gynecologists a better visualization of the format and the internal structures and in special the face of the embryo through the made process of three-dimensional reconstruction from a dataset of captured bi-dimensional images in conventional devices of ultrasound. The clinical uses of this project is foreseen will be the sector of obstetrics of the Hospital of the Clinics of Ribeirão Preto.

Par l’étude des représentations sociales, des rituels et des gestes funéraires qui entourent l’enfant dès sa naissance, cette thèse propose une réflexion sur la place des plus jeunes dans la société grecque archaïque et classique. Durant l’Antiquité, la venue au monde révèle les logiques sociales dans lesquelles elle s’inscrit : la mère est enfin reconnue comme épouse accomplie, l’homme devient père avec les implications sociales que cela entraîne, et la famille comme l’ensemble de la cité doit valider l’arrivée d’un nouveau membre. A partir de ce constat, ce travail apporte un nouveau regard sur les représentations de l’enfant en bas âge (0-3 ans) au sein des différentes strates de la cité. Longtemps considéré comme exclu des relations sociales, le tout-petit n’a en effet jamais fait l’objet d’une étude particulière. Pourtant des pratiques funéraires spécifiques, l’existence de rituels d’intégration progressifs, ainsi que des registres iconographiques distincts, sont autant de témoignages du statut particulier d’un être en devenir, mais déjà membre de la société. Ces différents thèmes sont abordés par une approche pluridisciplinaire qui confronte l’ensemble des sources disponibles sur la petite enfance : les écrits des auteurs anciens aux regards des imagiers et aux données offertes par l’archéologie funéraire. En conclusion ce travail montre que la notion de rite de passage vient s’ajouter aux représentations et aux pratiques funéraires pour former à un véritable complexe rituel faisant de la naissance et de la petite enfance, le reflet des rapports entretenus entre les Grecs et l’écoulement d’un temps inscrit dans la logique de reproduction sociale. / Through the study of social representations, rituals practices and funerary acts surrounding the young children, this thesis has offered a reflection on the infancy in the Archaic and Classical Greek world. During Antiquity, birth reveal the social mechanisms they belong to; the mother is finally considered as an accomplished wife and her role in the oikos is reaffirmed, the man becomes a father and has new social duties, and the family as well as the city has to accept the arrival of a new member. Judging from this observation, this work will shed a new light on young child (0-3 years) within the different strata of the city. For a long time, children have been considered as excluded from society because of the few mentions in literary sources. Although specific funerary practices, the existence of rituals marking the progressive integration of young ones in society, as well as a characteristic iconography, all testify of the particular status of children who already belong to the society. These themes are treated with a multidisciplinary approach that confronts all the sources on childhood: literature, iconography and funerary archaeology. In conclusion, this work proposes to show that the concept of rite of passage combined with representations and funerary practices form a ritual complex that makes birth and early childhood the true reflection of integration processes into the different circles of Greek society. This study thus aims to lead to a more general evocation of the relations between the Greeks of Antiquity and the passing of a time in the context of social reproduction.

In this 15 year follow up study in a Chinese population, we confirmed that maternal gestational diabetic status significantly increased women’s future cardiometabolic risk. Glycaemic levels below the current criteria for a positive screening test for gestational diabetes and for the diagnosis of gestational diabetes still significantly predict women’s future risk. In utero hyperinsulinaemia, which caused by an intrauterine hyperglycaemic environment, was found to predict children’s AGT and adolescents’ overweight and MetS. The results had important implication that the current diagnostic criteria for gestational diabetes may not be discriminative in predicting both the mothers and their children’s future cardiometabolic risk. Although recent research has re-visited and emphasised on the diagnostic criteria of gestational diabetes which best predicted adverse pregnancy outcome, future study should also scrutinise on the optimal glycaemic threshold, either in screening or diagnostic test, that relate to the mothers’ and children offspring’s long term cardiometabolic risk. / Tam, Wing Hung. / Thesis (M.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 119-146). / Abstract also in Chinese. / LIST OF TABLES --- p.xxii / LIST OF FIGURES --- p.xxv / LIST OF ABBREVIATIONS --- p.xxvi / Chapter Chapter 1 --- Gestational diabetes & future cardiometabolic risk - an overview / Chapter 1.1 --- Historical background --- p.2 / Chapter 1.2 --- Pregnancy physiology vs. gestational diabetes --- p.5 / Chapter 1.3 --- Diabetes mellitus - a global epidemic --- p.6 / Chapter 1.4 --- History of gestational diabetes & progression to Type 2 DM --- p.7 / Chapter 1.5 --- History of gestational diabetes & cardiometabolic risk --- p.8 / Chapter 1.6 --- Type 2 DM among children and adolescents --- p.9 / Chapter 1.7 --- Type 2 DM among offspring of mothers with gestational diabetes --- p.10 / Chapter 1.8 --- Cardiometabolic risk in children exposed to maternal gestational diabetes --- p.12 / Chapter 1.9 --- Long term follow up on mothers & children cohort --- p.12 / Chapter Chapter 2 --- Research methodology / Chapter 2.1 --- Subjects --- p.16 / Chapter 2.2 --- Obstetric and neonatal information --- p.18 / Chapter 2.2.1 --- Maternal glycaemic indices at pregnancy --- p.18 / Chapter 2.2.2 --- Umbilical cord blood C-peptide & insulin levels --- p.18 / Chapter 2.2.3 --- Definition of antenatal variables --- p.19 / Chapter 2.3 --- Follow up assessment of the mothers --- p.19 / Chapter 2.4 --- Follow up assessment of the children and adolescents --- p.22 / Chapter 2.5 --- Definition of abnormal glucose tolerance and metabolic syndrome --- p.24 / Chapter 2.5.1 --- Definition of abnormal glucose tolerance --- p.24 / Chapter 2.5.2 --- Definition of metabolic syndrome in adult --- p.24 / Chapter 2.5.3 --- Definition of metabolic syndrome in adolescent --- p.25 / Chapter 2.6 --- Determination of insulin resistance and pancreatic beta cell function --- p.26 / Chapter 2.6.1 --- Definition of insulin resistance --- p.26 / Chapter 2.6.2 --- Definition of pancreatic beta cell function --- p.26 / Chapter 2.6.3 --- Measurement of insulin resistance and pancreatic β-cell function --- p.27 / Chapter 2.7 --- Statistical analysis --- p.31 / Chapter 2.7.1 --- Statistical programme --- p.31 / Chapter 2.7.2 --- Comparison between group differences --- p.31 / Chapter 2.7.3 --- General Linear Model --- p.32 / Chapter 2.7.4 --- Multivariate logistic regression --- p.33 / Chapter 2.7.5 --- Receiver operating characteristic analysis --- p.37 / Chapter 2.8 --- Ethics approval --- p.41 / Chapter 2.9 --- Funding --- p.42 / Chapter Chapter 3 --- History of gestational diabetes and women’s future cardiometabolic risk / Chapter 3.1 --- Maternal clinical parameters at the index pregnancy --- p.44 / Chapter 3.2 --- Maternal cardiometabolic status at 8 years post-delivery --- p.45 / Chapter 3.3 --- Maternal cardiometabolic status at 15 years post-delivery --- p.49 / Chapter 3.4 --- Prediction of cardiometabolic risk by maternal gestational diabetic status --- p.50 / Chapter 3.4.1 --- Abnormal glucose tolerance and metabolic syndrome at 8 years by maternal gestational diabetic status --- p.52 / Chapter 3.4.2 --- Abnormal glucose tolerance, DM, hypertension and metabolic syndrome at 15 years by maternal gestational diabetic status --- p.52 / Chapter 3.5 --- The role of insulin resistance in predicting women’s DM and metabolic syndrome --- p.55 / Chapter 3.6 --- Discussion --- p.57 / Chapter 3.7 --- Conclusion --- p.62 / Chapter Chapter 4 --- Glycaemic variables measured at mid-gestation of the index pregnancy predict women’s future cardiometabolic risk / Chapter 4.1 --- Glycaemic levels in pregnancy and perinatal outcome --- p.64 / Chapter 4.2 --- Glycaemic levels in pregnancy and women’s future cardiometabolic risk --- p.65 / Chapter 4.2.1 --- Prediction of women’s cardiometabolic risk at 8 and 15-year --- p.66 / Chapter 4.2.2 --- Optimal cut-off levels in predicting women’s future cardio- metabolic risk --- p.69 / Chapter 4.3 --- Discussion --- p.75 / Chapter 4.4 --- Conclusion --- p.78 / Chapter Chapter 5 --- Maternal gestational diabetes and offspring’s cardiometabolic risk / Chapter 5.1 --- Offspring’s cardiometabolic risk at 8 years age --- p.80 / Chapter 5.1.1 --- Baseline characteristics at pregnancy and delivery --- p.80 / Chapter 5.1.2 --- Children’s clinical and biochemical parameters at 8 years age --- p.82 / Chapter 5.2 --- Offspring’s cardiometabolic risk at 15 years age --- p.84 / Chapter 5.2.1 --- Adolescents’ clinical and biochemical parameters at 15 years age --- p.84 / Chapter 5.2.2 --- Clinical parameters of adolescents with abnormal glucose tolerance --- p.84 / Chapter 5.3 --- Discussion --- p.88 / Chapter 5.4 --- Conclusion --- p.90 / Chapter Chapter 6 --- In utero hyperinsulinaemia and offspring’s cardiometabolic risk / Chapter 6.1 --- Umbilical cord blood insulin and C-peptide --- p.92 / Chapter 6.1.1 --- Umbilical cord insulin and C-peptide concentrations in the original cohort --- p.92 / Chapter 6.1.2 --- Determination of in utero hyperinsulinaemia by umbilical cord insulin and C-peptide levels --- p.95 / Chapter 6.2 --- The effect of in utero hyperinsulinaemia on children’s abnormal glucose tolerance at 8 years of age --- p.98 / Chapter 6.2.1 --- Receiver operating characteristic analysis --- p.98 / Chapter 6.2.2 --- Logistic regression analysis --- p.98 / Chapter 6.3 --- The effect of in utero hyperinsulinaemia on adolescents’ cardio- metabolic risk at 15years of age --- p.102 / Chapter 6.3.1 --- Logistic regression analysis --- p.102 / Chapter 6.4 --- Discussion --- p.105 / Chapter 6.5 --- Conclusion --- p.108 / Chapter Chapter 7 --- Summary and conclusion / Chapter 7.1 --- Summary of the thesis --- p.110 / Chapter 7.1.1 --- Women’s long term cardiometabolic risk after a pregnancy with gestational diabetes --- p.110 / Chapter 7.1.2 --- The long term cardiometabolic risk of children born to mothers who had gestational diabetes --- p.111 / Chapter 7.1.3 --- New findings from the studies and their implications --- p.111 / Chapter 7.2 --- Strength and weakness in the study --- p.113 / Chapter 7.2.1 --- Unique cohort from universal screening --- p.113 / Chapter 7.2.2 --- Study design --- p.113 / Chapter 7.2.3 --- Response rate and loss to follow up --- p.114 / Chapter 7.2.4 --- Treatment effect of gestational diabetes --- p.115 / Chapter 7.3 --- Issues of future research --- p.115 / Chapter 7.3.1 --- Follow up study on the HAPO cohort --- p.115 / Chapter 7.3.2 --- Opportunity for international collaboration --- p.117 / Chapter 7.4 --- Conclusion --- p.118 / REFERENCES --- p.119

INTRODUÇÃO: A função renal fetal é avaliada tradicionalmente pelo aspecto ultrassonográfico, pela análise bioquímica da urina fetal, somada com o diagnóstico exato da natureza da lesão. A análise bioquímica da urina é uma avaliação invasiva e apresenta riscos maternos e fetais. A avaliação por meio da ultrassonografia é método não invasivo e a presença de oligoâmnio é indicativo de displasia renal grave, contudo, de diagnóstico tardio nos casos de patologia renal fetal. O volume renal fetal pode correlacionar-se com a sua função e pode ser avaliado por meio da ultrassonografia bidimensional. Entretanto, esse método apresenta subestimação no cálculo de volume, enquanto por meio da ultrassonografia tridimensional há maior precisão e acurácia em avaliar o volume renal fetal. A ultrassonografia Power Doppler tridimensional é o exame ideal para avaliar a vascularização de parênquima de órgãos fetais e pode ser útil na avaliação da função do rim fetal. OBJETIVOS: O presente estudo tem por objetivos utilizar a ultrassonografia tridimensional acoplado ao Power Doppler e elaborar curvas com valores de referência do volume e dos índices de vascularização dos rins fetais segundo a idade gestacional. MÉTODOS: Realizou-se estudo prospectivo e transversal em gestantes sem patologia, com fetos únicos normais, entre 20 semanas completas a 39 semanas e 6 dias, que foram avaliados por meio da ultrassonografia tridimensional acoplada ao Power Doppler. Foram calculados o volume e os índices de vascularização dos rins fetais, construindo-se as curvas de normalidade. Os testes foram realizados com nível de significância de 5%. A variação intra e interobservador também foi analisada. RESULTADOS: Duzentos e treze fetos foram analisados, sendo que, em 211 casos, analisou-se o rim direito; em 209 o rim esquerdo e, em 204, os índices de vascularização. Os resultados foram ilustrados em gráficos de dispersão com os respectivos intervalos de normalidade com os percentis 5, 10, 25, 50, 75, 90 e 95 e criou-se uma tabela com as equações para estimar as medidas dos volumes renais e dos índices de vascularização pelos modelos de regressão. A avaliação intra e interobservador das mensurações do volume e dos índices de vascularização dos rins fetais apresentou boa reprodutibilidade. CONCLUSÃO: As medidas dos volumes renais fetais variam de forma exponencial, ao longo da idade gestacional, de acordo com os valores de referência propostos e as medidas dos índices de vascularização renais fetais, exceto o índice de fluxo, variam também de forma exponencial, ao longo da idade gestacional, de acordo com os valores de referência apresentados / INTRODUCTION: The fetal renal function is traditionally evaluated by ultrasound aspect, by biochemical analysis of fetal urine together with an accurate diagnosis of the nature of the lesion. Biochemical analysis of the urine is a invasive evaluation and presents risks maternal and fetal. The evaluation by means of ultrasound is noninvasive method and the presence of oligohydramnios is indicative of renal dysplasia serious, however of late diagnosis in cases of renal pathology fetal. The volume fetal renal can correlate with its function and may be assessed by means of twodimensional ultrasound. However, this method presents an underestimation in the calculation of volume, already by means of three-dimensional ultrasonography shows greater precision and accuracy in assessing the volume fetal renal. The threedimensional Power Doppler ultrasonography is the ideal examination to evaluate the vascularization of the parenchyma of fetal organs and may be useful in the evaluation of the function of the fetal kidney. OBJECTIVES: This study aims to use threedimensional ultrasonography coupled to Power Doppler and elaborate curves with reference values of the volume and indices of vascularization of fetal kidneys according to the gestational age. METHODS: A prospective study was performed and cross-sectional in pregnant women without pathology with fetuses only normal between 20 full weeks to 39 weeks and 6 days that were evaluated by means of three-dimensional ultrasonography coupled to Power Doppler. We calculated the volume and the index of vascularization of fetal kidneys, built to the curves of normality. The tests were performed with significance level of 5 %. The variation intra and interobserver were also analyzed. RESULTS: Two hundred and thirteen fetuses were analyzed, in 211 cases examined whether the right kidney; in 209, the left kidney and in 204 the indexes of vascularization. The results were illustrated in dispersion graphs with their normal ranges with percentiles 5, 10, 25, 50, 75, 90 and 95 and created a table with the equations to estimate measures of renal volume and indices of vascularization by the regression models. To assess intra and interobserver measurements of volume and indices of vascularization of fetal kidneys showed good reproducibility. CONCLUSION: The measures of renal volume fetal vary exponentially, along the gestational age, of agreement with the values of reference proposed and the measures of the indices of vascularization fetal renal, except the flow index, also vary exponentially, along the gestational age, of agreement with the reference values presented

Objetivo: Caracterizar as indicações das gestantes que procuram o serviço de Medicina Fetal do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo que realizaram procedimentos invasivos diagnósticos e avaliar os resultados dos cariótipos fetais e destas gestações no período de fevereiro de 2005 a dezembro de 2009. Método: Estudo observacional retrospectivo das gestantes que realizaram biópsia de vilo corial (BVC), amniocentese e cordocentese. Não foram incluídos outros procedimentos diagnósticos ou terapêuticos. O resultado da gestação foi obtido através de consulta de prontuário eletrônico e/ou físico e/ou contato telefônico. Resultados: Foram realizados 113 BVC, 340 amniocenteses e 260 cordocenteses. A principal indicação para a realização dos procedimentos invasivos foi a presença de malformações fetais (69,8%), seguido por translucência nucal aumentada (13,4%) e idade materna avançada (10,2%). A trissomia do cromossomo 18 foi a aneuploidia mais comum (8,1%), seguido pela trissomia do 21 (6,2%), 45,X0 (4,8%) e a trissomia do 13 (3,8%). Ocorreram 4,9% abortamentos, 25,7% natimortos e 13% neomortos. Oito gestantes optaram pela interrupção judicial. 99% das gestantes cujos fetos não apresentavam malformação e que apresentavam cariótipo fetal normal tiveram nativivos. CONCLUSÃO: A principal indicação para a realização dos procedimentos invasivos foi a presença de malformação fetal em 69,8% das gestantes e presença de anormalidades cromossômicas encontradas nos fetos foi de 26,23%. / Objective: The purpose of this study is to characterize the indications of pregnant women who seek the Fetal Medicine Service of Hospital das Clínicas of São Paulo University to perform invasive diagnostic procedures and evaluate the results of fetal karyotypes and their pregnancies from February 2005 to December 2009. Methods: Retrospective observational study of pregnant women who underwent CVS, amniocentesis or cordocentesis. Other diagnostic or therapeutic procedures were not included. The outcomes of pregnancies were obtained through consultation of medical records and/or telephone contact. Results: 113 CVS, 340 amniocentesis and 260 cordocentesis were performed. The main indication for performing invasive procedure was the presence of fetal anomaly (69.8%), followed by increased nuchal translucency (13.4%) and maternal age (10.2%). The trisomy of chromosome 18 was the most common aneuploidy (8.1%), followed by trisomy 21 (6.2%), 45,X0 (4.8%), and trisomy 13 (3.8%). There were 4.9 % of miscarriage, 25.7% of stillbirth and 13% of neonatal deaths. Eight women opted for legal termination of pregnancies. 99% of pregnant women whose fetus had no structural abnormalities and normal karyotype had a live child. CONCLUSION: The main indication for karyotyping was the presence of fetal malformation in 69.8% of pregnancies and chromosomal abnormalities was found in 26.23% of the fetuses.

Made available in DSpace on 2016-04-28T20:18:36Z (GMT). No. of bitstreams: 1 Viviane de Souza Cardoso.pdf: 1798596 bytes, checksum: 2862a717209bf03e8568e17d21be6ba4 (MD5) Previous issue date: 2003-02-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The headquartered fractures in the appendixes of the small animals represent great portion of the service in surgical clinics. The purpose of this work was evaluate the bone regeneration with the use of allografts or bovine fetus xenografts, conserved in solution of cefalotina sodic neutral to 0,5%. Twenty adult mongrel dogs were used. We done ostectomia of a fragment of the tibia of the receptor dogs with dimensions same to 10mm x 4mm where then in its place, it was put the allograft or xenograft, in agreement with the group to which belonged the animal. Clinically, support of the limb was observed in the subsequent day to the surgery and general state of the animal. The radiographic evaluation was accomplished before the surgical procedure and in the postoperative immediate, on the 15, 30, 45 and 60 days of evolution. In 15 days beginning of formation of bone callus was observed in the allograft group, already in the xenograft group this was observed to the 30 days in almost all the animals. There were no evidence infection, of deformity or any other alteration in the operated timb, and we concluded that the allograft is incorporated faster than the xenograft, however not being discarded the use possibility of this use. / As fraturas sediadas nos ap?ndices locomotores dos pequenos animais representam grande parcela do atendimento em cl?nicas cir?rgicas. O objetivo deste trabalho foi avaliar a regenera??o ?ssea com a utiliza??o de aloimplantes e de xenoimplantes de feto bovino em c?o, conservados em solu??o de cefalotina s?dica tamponada a 0,5%. Foram utilizados 20 c?es h?gidos, adultos, sem ra?a definida e de m?dio porte. Realizou-se ostectomia de um fragmento da t?bia dos c?es receptores de dimens?es iguais a 10mm x 4mm onde ent?o em seu lugar foi colocado o implante al?geno ou xen?geno, de acordo com o grupo ao qual pertencia o animal. Clinicamente, observou-se apoio do ap?ndice operado no dia posterior ao procedimento e estado geral do animal. A avalia??o radiogr?fica foi realizada antes do procedimento cir?rgico e no p?s-operat?rio imediato, aos 15, 30, 45 e 60 dias de evolu??o. Aos 15 dias observou-se in?cio de forma??o de calo ?sseo no grupo aloimplante, j? no grupo xenoimplante este foi observado aos 30 dias em quase todos os animais. N?o houve evid?ncia de infec??o, deformidade ou qualquer outra altera??o nos ap?ndices operados, e comparando-se os m?todos concluise que o aloimplante ? incorporado com maior velocidade que o xenoimplante, por?m n?o sendo descartada a possibilidade de utiliza??o deste.

Prenatal diagnosis is an established obstetrics practice in many countries. Currently available methods to obtain fetal materials for a definitive diagnosis involve invasive procedures such as chorionic villus sampling and amniocentesis. Due to the invasive nature of the procedures, confirmatory testing is usually recommended only for pregnant women who are screened as being high risk of bearing a fetus with abnormalities. There has been an urge to develop safer alternatives in obtaining fetal genetic materials for prenatal assessment. Thus, the discovery of circulating fetal DNA in maternal plasma and serum has opened up new possibilities for noninvasive prenatal diagnosis. / The use of genetic markers such as Y chromosomal sequences from a male fetus or paternally-inherited polymorphic markers has been well-described in the literature. However, there is an obvious limitation on the use of these markers because they are gender- and/or polymorphism-dependent. This thesis focuses on the development of a universal fetal marker, namely SERPINB5 (encoding maspin), based on the intrinsic epigenetic differences between the placenta (the major contributor of fetal genetic materials in maternal plasma) and maternal blood cells (postulated to be the predominant source of cell-free DNA in the circulation). Analysis of the methylation profile of the SERPINB5 promoter in the placenta and maternal blood cells, and the development of methods and protocols to detect the differentially methylated SERPINB5 promoter molecules are described. The application of this epigenetic marker in prenatal assessment of the fetal chromosomal aneuploidy is illustrated by an epigenetic allelic ratio (EAR) approach. Basically, the ratio of a single-nucleotide polymorphism (SNP) on the placenta-derived SERPINB5 molecules is shown to be deviated in the maternal plasma from trisomic pregnancies when compared to the euploid ones. Results described in this thesis show the promising potential for the EAR approach for the noninvasive prenatal detection of fetal chromosomal aneuploidies. In addition, a novel approach for methylation analyses on the amplification and detection of restriction enzyme-digested DNA fragments will be discussed. This thesis has essentially described the evolution of a fetal epigenetic marker from basic science to potential clinical application. / Tong, Yu Kwan. / Adviser: Yuk-Ming Dennis Lo. / Source: Dissertation Abstracts International, Volume: 69-02, Section: B, page: 0953. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 149-172). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / School code: 1307.

Pang, Weng I. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 116-148). / Abstracts in English and Chinese. / ABSTRACT --- p.i / 摘要 --- p.iv / ACKNOWLEDGEMENTS --- p.vi / PUBLICATIONS --- p.vii / TABLE OF CONTENTS --- p.viii / LIST OF TABLES --- p.xiii / LIST OF FIGURES --- p.xv / LIST OF ABBREVIATIONS --- p.xvi / Chapter SECTION I: --- BACKGROUND --- p.1 / Chapter CHAPTER 1: --- CIRCULATING NUCLEIC ACIDS IN PRENATAL DIAGNOSIS --- p.2 / Chapter 1.1 --- Prenatal diagnosis --- p.2 / Chapter 1.2 --- Circulating fetal DNA in maternal plasma --- p.2 / Chapter 1.2.1 --- Biology of circulating fetal DNA --- p.2 / Chapter 1.2.2 --- Clinical applications of circulating fetal DNA --- p.3 / Chapter 1.2.2.1 --- Qualitative fetal-specific sequence detection --- p.4 / Chapter 1.2.2.2 --- Quantitative aberration detection --- p.4 / Chapter 1.2.3 --- Circulating fetal epigenetic markers --- p.5 / Chapter 1.3 --- Circulating fetal RNA in maternal plasma --- p.6 / Chapter 1.3.1 --- Biology of circulating fetal RNA --- p.6 / Chapter 1.3.2 --- Clinical applications of circulating fetal RNA --- p.8 / Chapter 1.3.2.1 --- Quantitative aberration detection --- p.8 / Chapter 1.3.2.2 --- Chromosomal aneuploidy detection --- p.9 / Chapter 1.3.3 --- Enrichment of fetal RNA --- p.10 / Chapter 1.4 --- Circulating microRNA in maternal plasma --- p.10 / Chapter CHAPTER 2: --- FETAL GROWTH AND WELL-BEING --- p.12 / Chapter 2.1 --- Normal fetal growth --- p.12 / Chapter 2.1.1 --- Role of the mother --- p.12 / Chapter 2.1.2 --- Role of the placenta --- p.12 / Chapter 2.1.3 --- Role of the fetus --- p.13 / Chapter 2.1.4 --- Role of the somatotrophic axis --- p.15 / Chapter 2.2 --- Abnormal fetal growth --- p.15 / Chapter 2.2.1 --- Intrauterine growth restriction --- p.16 / Chapter 2.1.2 --- Definition of IUGR --- p.16 / Chapter 2.2.3 --- Risk factors of IUGR --- p.17 / Chapter 2.2.4 --- Diagnosis of IUGR --- p.20 / Chapter 2.2.4.1 --- Biometric tests --- p.20 / Chapter 2.2.4.2 --- Biophysical tests --- p.21 / Chapter 2.2.4.3 --- Biochemical tests --- p.22 / Chapter 2.2.4.4 --- Others --- p.22 / Chapter 2.3 --- Limitations of current modalities in fetal growth assessment --- p.23 / Chapter 2.4 --- Aims of this thesis --- p.24 / Chapter SECTION II: --- MATERIALS AND METHODS --- p.26 / Chapter CHAPTER 3: --- QUANTITATIVE ANALYSIS OF CIRCULATING RNA --- p.27 / Chapter 3.1 --- Sample collection and processing --- p.27 / Chapter 3.1.1 --- Preparation of plasm a --- p.27 / Chapter 3.1.2 --- Preparation of blood cells --- p.27 / Chapter 3.1.3 --- Preparation of placental tissues --- p.27 / Chapter 3.2 --- Total RNA extraction --- p.28 / Chapter 3.2.1 --- Plasma and blood cells --- p.28 / Chapter 3.2.2 --- Placental tissues --- p.32 / Chapter 3.3 --- Quantitative measurements of nucleic acids --- p.32 / Chapter 3.3.1 --- Principles of real-time quantitative PCR --- p.33 / Chapter 3.3.2 --- One-step QR T-PCR assays for placental mRNA quantification --- p.3 7 / Chapter 3.3.3 --- QPCR assays for checking genomic DNA contamination --- p.43 / Chapter 3.4 --- Statistical analysis --- p.45 / Chapter SECTION III: --- EVALUATION OF PLACENTA-DERIVED MRNA AS POSSIBLE MARKERS FOR FETAL GROWTH ASSESSMENT --- p.46 / Chapter CHAPTER 4: --- SELECTION OF POTENTIAL FETAL GROWTH MRNA MARKERS FOR MATERNAL PLASMA DETECTION --- p.47 / Chapter 4.1 --- Introduction --- p.47 / Chapter 4.2 --- Materials and methods --- p.49 / Chapter 4.2.1 --- Sample collection and processing --- p.49 / Chapter 4.2.2 --- Experimental design --- p.49 / Chapter 4.2.3 --- RNA extraction and quantification --- p.51 / Chapter 4.2.4 --- Statistical analysis --- p.51 / Chapter 4.3 --- Results --- p.52 / Chapter 4.3.1 --- Identification of potential fetal growth mRNA markers in maternal plasma --- p.52 / Chapter 4.3.2 --- Development of real-time QR T-PCR assays --- p.56 / Chapter 4.3.3 --- Validation of maternal plasma detectability and pregnancy-specificity --- p.58 / Chapter 4.3.4 --- Assessment of the gestational trend in maternal plasma --- p.64 / Chapter 4.4 --- Discussion --- p.68 / Chapter CHAPTER 5: --- RELATIONSHIP BETWEEN CIRCULATING PLACENTAL MRNA AND FETAL GROWTH --- p.72 / Chapter 5.1 --- Introduction --- p.72 / Chapter 5.2 --- Materials and methods --- p.73 / Chapter 5.2.1 --- Sample collection and processing --- p.73 / Chapter 5.2.2 --- "Ultrasound measurement, placental weight and birth weight.…" --- p.74 / Chapter 5.2.3 --- Experimental design --- p.74 / Chapter 5.2.4 --- RNA extraction and quantification --- p.75 / Chapter 5.2.5 --- Statistical analysis --- p.75 / Chapter 5.3 --- Results --- p.75 / Chapter 5.3.1 --- Expression of potential growth markers in placental tissues --- p.76 / Chapter 5.3.2 --- Relationship between circulating placental mRNA and birth measurements --- p.76 / Chapter 5.3.3 --- Relationship between circulating placental mRNA and fetal biometric measurements --- p.77 / Chapter 5.4 --- Discussion --- p.85 / Chapter SECTION IV: --- CLINICAL APPLICATION OF POTENTIAL FETAL GROWTH MARKERS IN THE ASSESSMENT OF IUGR --- p.93 / Chapter CHAPTER 6: --- QUANTITATIVE ANALYSIS OF PLACENTAL MRNA IN IUGR WITH OR WITHOUT PET --- p.94 / Chapter 6.1 --- Introduction --- p.94 / Chapter 6.2 --- Materials and methods --- p.95 / Chapter 6.2.1 --- Sample collection and processing --- p.95 / Chapter 6.2.2 --- Experimental design --- p.96 / Chapter 6.2.3 --- RNA extraction and quantification --- p.96 / Chapter 6.2.4 --- Statistical analysis --- p.97 / Chapter 6.3 --- Results --- p.97 / Chapter 6.3.1 --- Cross-sectional comparison of placental mRNA concentrations --- p.97 / Chapter 6.3.2 --- Longitudinal comparison of placental mRNA concentrations --- p.102 / Chapter 6.4 --- Discussion --- p.103 / Chapter SECTION V: --- CONCLUDING REMARKS --- p.107 / Chapter CHAPTER 7: --- CONCLUSION AND FUTURE PERSPECTIVES --- p.108 / Chapter 7.1 --- A strategy for identifying circulating placental MRNA markers for fetal growth assessment --- p.108 / Chapter 7.2 --- Implications of mRNA marker development strategy --- p.111 / Chapter 7.3 --- Prospects for future work --- p.112 / REFERENCES --- p.116

Fetal growth restriction (FGR) is a common and potentially life-threatening complication that affects 5-10% of human pregnancies. Maternal genetic predisposition and fetal male sex are known risk factors, but the underlying mechanisms are unknown. To study a known maternal genetic risk factor and the impact of fetal sex, we employed a published transgenic (TG) mouse model, which was designed to mimic a common human angiotensinogen (AGT) promoter variant associated with a 20% increase in circulating AGT levels. We hypothesized that TG dams would deliver growth restricted pups and that the underlying mechanism would be related to differences in maternal uterine pregnancy-induced vascular remodeling, abnormal blood flow to the placenta, and placental damage. In addition, since growth restricted human males are at an increased risk of developing adult onset hypertension, which has been associated with reduced nephron development, we tested for developmental programming in our mouse model and the impact of fetal sex. Our results show that TG dams have reduced uterine and placental angiogenesis when their pups were males, but relatively normal angiogenesis in the female siblings compared with wild-type controls. The uterine placental bed in TG dams had abnormal pro-angiogenic/anti-angiogenic expression ratios that were related to differences in uterine natural killer cell activation and fetal sex. The abnormal phenotype could be rescued by delivering vascular endothelial growth factor (VEGF) to uterine endothelial cells. Male progeny from TG dams had abnormal kidney epigenetic changes, fewer nephrons as adults, and they developed stress-induced hypertension. We conclude that the combination of maternal genetic risk and fetal male sex affect uteroplacental angiogenesis leading to FGR and the programming of stress-induced hypertension.

Abstract not available