Study questions: Does the proportion of neural tube defects (NTDs) in which there are co-occurring anomalies of other systems vary by geographical area and time in Europe between 1980 and 2015? Which group of factors, e.g., sex, age, family history, condition at discovery, and prenatal screening during pregnancy that potentially affect ascertainment of isolated NTD cases and co-occurring anomalies, are associated with the presence of these anomalies? Methods: Data on fetuses or births with NTDs recorded in 28 European Surveillance of Congenital Anomalies (EUROCAT) registries operating throughout or during a part of the period 1980-2015, were obtained and analysed. Information on the corresponding denominators of total births for each registry were compiled, comprising approximately 15.7 million births. We developed algorithms to classify NTDs that occurred as part of certain recognized genetic and unknown etiology syndromes, isolated NTD cases (either with no other anomaly reported or only an anomaly considered to be secondary to the NTD), and NTDs not occurring as part of recognized syndromes but with one or more co-occurring anomalies. We assessed factors that could impact ascertainment of isolated NTD cases and NTD cases with co-occurring anomalies. We also conducted a sensitivity analysis to evaluate the effect of family history and consanguinity on NTD outcomes. Lastly, we reported on the prevalence of NTDs across Eastern, Western, Northern, and Southern Europe as defined by the World Bank. Results: A total of 14,703 fetuses or births with NTDs were recorded in the EUROCAT registries. The total number of births (live and stillbirths) that occurred during the periods when the EUROCAT registries were operating was 15.7 million. The overall prevalence at birth of NTDs in Eastern Europe was 11.6 per 10,000 births, in Western Europe 11.0 per 10,000 births, in Northern Europe 10.5 cases per 10,000 births and in Southern Europe 6.7 per 10,000 births. The most prevalent genetic syndromes were chromosomal anomalies, particularly trisomy 18. The most common secondary anomaly was Arnold-Chiari syndrome which occurred mostly with spina bifida. The most prevalent associated anomalies were limb defects, followed by congenital heart defects, abdominal wall, and urinary system defects. We found anencephaly to be most prevalent in isolated NTD cases without secondary anomalies and spina bifida in secondary anomalies and most of the co-occurring anomalies. For factors that could affect ascertainment, we found that birth type, birth weight, family history, maternal and paternal age (≥30 years) were risk factors for isolated NTD cases and for co-occurring anomalies. Lastly, approximately 48% of NTDs occurred in Northern Europe, followed 29% in Western Europe, 16% in Southern Europe, and 7.6% in Eastern Europe. Conclusion: In the absence of folate fortification, isolated NTD cases showed higher prevalence than NTDs with co-occurring anomalies over the past 35 years across Europe. This research suggests that Europe should develop and implement policies and guidelines for folate fortification to help prevent simpler NTD cases. Further research and data are needed for newer EUROCAT registries to predict trends and prevalence of NTDs. Contribution and involvement: This project was done in collaboration with the experts from the School of Epidemiology and Public Health (SEPH) and dysmorphologists that provided support with regards to NTD classification. AB spearheaded the development of the research protocol and thesis manuscript with funding and support from JL and TR. Evaluation of the output of cases from the algorithm was provided to AB by JL, TR, JE, FB, AN, EB, ML, and the EUROCAT Joint Research Centre.

No description available.

La voie de la polarité planaire cellulaire (PCP), aussi connue sous le nom de la voie non-canonique du Frizzled/Dishevelled, contrôle le processus morphogénétique de l'extension convergente (CE) qui est essentiel pour la gastrulation et la formation du tube neural pendant l'embryogenèse. La signalisation du PCP a été récemment associée avec des anomalies du tube neural (ATN) dans des modèles animaux et chez l'humain. Prickle1 est une protéine centrale de la voie PCP, exprimée dans la ligne primitive et le mésoderme pendant l'embryogenèse de la souris. La perte ou le gain de fonction de Prickle1 mène à des mouvements de CE fautifs chez le poisson zèbre et la grenouille. PRICKLE1 interagit directement avec deux autres membres de la voie PCP, Dishevelled et Strabismus/Vang. Dans notre étude, nous avons investigué le rôle de PRICKLE1 dans l'étiologie des ATN dans une cohorte de 810 patients par le re-séquençage de son cadre de lecture et des jonctions exon-intron. Le potentiel pathogénique des mutations ainsi identifiées a été évalué par des méthodes bioinformatiques, suivi par une validation fonctionnelle in vivo dans un système poisson zèbre. Nous avons identifié dans notre cohorte un total de 9 nouvelles mutations dont sept: p.Ile69Thr, p.Asn81His, p.Thr275Met, p.Arg682Cys et p.Ser739Phe, p.Val550Met et p.Asp771Asn qui affectent des acides aminés conservés. Ces mutations ont été prédites in silico d’affecter la fonction de la protéine et sont absentes dans une large cohorte de contrôles de même origine ethnique. La co-injection de ces variantes avec le gène prickle1a de type sauvage chez l’embryon de poisson zèbre a démontré qu’une mutation, p.Arg682Cys, modifie dans un sens négatif le phénotype du défaut de la CE produit par pk1 de type sauvage. Notre étude démontre que PK1 peut agir comme facteur prédisposant pour les ATN chez l’humain et élargit encore plus nos connaissances sur le rôle des gènes de la PCP dans la pathogenèse de ces malformations. / The planar cell polarity pathway (PCP) or the non-canonical Frizzled/Dishevelled pathway controls the morphogenetic process of convergent extension (CE) that is essential during embryogenesis for gastrulation and neural tube formation. Recently, PCP signalling was associated with neural tube defects (NTD) in humans and animal models. The core PCP protein, Prickle1, is expressed in the primitive streak and mesoderm during mouse embryogenesis. Both gain and loss of function of Prickle1 cause faulty CE movements in zebrafish and the frog. PRICKLE1 physically interacts with two other core PCP members, Dishevelled and Strabismus/Vang. In the present study we investigated the role of PRICKLE1 in the aetiology of NTDs in a large cohort of 810 patients through resequencing of its open reading frame and exon-intron junctions. The pathogenicity of the identified mutations was assessed through bioinformatics methods followed by a functional validation in a zebrafish system, in vivo. We identified in our cohort a total of nine novel mutations, of which seven affected conserved amino acids: p.Ile69Thr, p.Asn81His, p.Thr275Met, p.Arg682Cys, p.Ser739Phe, p.Val550Met and p.Asp771As. These mutations were predicted to affect the function of the protein in silico and were absent in a large cohort of ethnically-matched controls. Co-injection of these variants with the wild type pk1 in zebrafish oocytes revealed that one mutation, p.Arg682Cys, antagonized the CE phenotype induced by the wild-type zebrafish prickle1a in a dominant fashion. Our study demonstrates that PRICKLE1 can represent a predisposing factor for human NTDs and further expands our knowledge on the role that PCP genes in the pathogenesis of these malformations.

Les anomalies du tube neural (ATN), incluant l'anencéphalie et le spina-bifida, représentent un groupe de malformations congénitales très fréquentes chez l'homme. Ces anomalies sont causées par un défaut partiel ou complet de la fermeture du tube neurale au cours de l'embryogenèse. Les ATN ont une étiologie complexe et multifactorielle impliquant des facteurs environnementaux et génétiques. La voie de signalisation non-canonique du Frizzled (Fz)/Dishevelled (Dvl) contrôle la polarité cellulaire planaire (PCP) et le processus morphogénétique appelé l’extension convergente qui est essentiel pour la gastrulation et la fermeture du tube neural. Très important, des mutations des gènes de cette voie étaient fortement associées aux ATN chez la souris et l’humain. Scribble est un gène de la voie PCP qui cause une sévère ATN chez la souris Circletail. Notre étude vise à analyser le rôle de SCRIBBLE1 dans les ATN humains par des analyses de séquence de son cadre de lecture et ses jonctions exon-introns. Notre étude comporte 396 patients recrutés au Centre Spina Bifida de l’hôpital Gaslini en Gènes, Italie et 83 patients recrutés au Centre Spina Bifida de l’hôpital Sainte Justine. Les patients sont affectés par plusieurs formes d’ATN. Nous avons identifié neuf mutations rares et non synonymes chez 10 patients, p.Asp93Ala (c. 435G>A), p.Gly145Arg (c. 278A>C), p.Gly263Ser (c. 786C>A), p.Gly469Ser (c. 1405G>A), p.Pro649His (c. 1946C>A), p.Gln808His (c. 2424G>T), p.Val1066Met (c. 3196G>A), p.Arg1150Gln (c. 3480G>A) et p.Thr1422Met (c. 4266C>T). Cinque mutations, p.Gly263Ser, p.Pro649His, p.Gln808His, p.Arg1150Gln, p.Thr1422Met, étaient absentes dans les contrôles analysés et prédites d’être pathogéniques in silico. Cette étude montre que des mutations rares dans SCRIB1 pourraient augmenter le risque des ATN dans une fraction des patients. L’identification des gènes prédisposant aux ATN nous aidera à mieux comprendre les mécanismes pathogéniques impliqués dans ces maladies. / Neural tube defects (NTDs), including anencephaly and spina bifida, represent a group of very common birth defects in humans. These anomalies are caused by a partial or complete failure of neural tube closure during embryogenesis. NTDs have a multifactorial etiology involving environmental and genetic factors. The non-canonical signaling pathway Frizzled (Fz) / Dishevelled (Dvl) controls the planar cell polarity (PCP) and the morphogenetic process called convergent extension (CE) which is essential for gastrulation and neural tube closure. Importantly, mutations in genes of this pathway were strongly associated with NTDs in mice and humans. Scribble is a PCP gene that causes a severe NTD mouse Circletail. Scribble binds to another PCP protein, Stbm / Vang, and they cooperate together for the stability of the PCP pathway. Our study aims at investigating the role of SCRIBBLE1 in human NTDs by sequence analyses of its open reading frame and exon-intron junctions. The cohort included in this study consisted of 396 patients recruited at the Spina Bifida Centre of Gaslini Hospital in Genoa, Italy, and 83 patients recruited at the Spina Bifida Center of the Sainte Justine Hospital, Montreal, Canada. Patients were affected by several forms of NTDs. We identified nine non-synonymous and rare mutations in 10 patients: p.Asp93Ala (c. 435G>A), p.Gly145Arg (c. 278A>C), p.Gly263Ser (c. 786C>A), p.Gly469Ser (c. 1405G>A), p.Pro649His (c. 1946C>A), p.Gln808His (c. 2424G>T), p.Val1066Met (c. 3196G>A), p.Arg1150Gln (c. 3480G>A) and p.Thr1422Met (c. 4266C>T). Five of those mutations, p.Gly263Ser, p.Pro649His, p.Gln808His, p.Arg1150Gln, p.Thr1422Met, were absent in all controls analyzed and were predicted to be pathogenic using bioinformatics. Our study demonstrates that rare mutations in SCRIB1 could predispose to NTDs in a fraction of patients. The identification of genes that predispose to ATN will help us better understand the pathogenic mechanisms involved in these diseases.

Made available in DSpace on 2016-03-22T17:27:38Z (GMT). No. of bitstreams: 1 cintia.pdf: 399237 bytes, checksum: 411794e4a2f292dd1123c42f8b410d51 (MD5) Previous issue date: 2006-11-20 / To determine folic acid use and associated factors, a cross-sectional population-based study was carried out in the five maternities from Pelotas - RS, Brazil. Data were collected from April 1st to August 15th, 2006. 1450 women were interviewed. A standard questionnaire was applied in the hospitals. Statistical analysis was done by Poisson Regression. Prevalence of folic acid consumption during pregnancy was 31.8% and periconceptional use was 4.3%. The following characteristics were associated to folic acid use: white skin color, schooling above 9 years, family income higher than 600 reais, age groups older than 30, planned pregnancies, seven or more prenatal care visits, knowledge about folic acid and prenatal care realization in the private health system. It is important to promote folic acid use among fertile age women and to supply with folic acid poorer women in order to prevent neural tube defects / Com o objetivo de determinar a prevalência do uso do ácido fólico e fatores associados, realizou-se um estudo transversal de base populacional nas cinco maternidades da cidade de Pelotas - RS, Brasil. A coleta de dados ocorreu no período de 1° de abril a 15 de agosto de 2006 com 1.450 mulheres. As entrevistas foram realizadas em nível hospitalar por questionário padronizado. A análise estatística se realizou por regressão de Poisson. A prevalência do uso de ácido fólico na gestação foi de 31,8%, e no período periconcepcional, foi de 4,3%. Os fatores associados ao uso de ácido fólico foram: cor branca, escolaridade acima de nove anos, renda acima de 600 reais, idade acima de 30 anos, gestação planejada, sete ou mais consultas de pré-natal, consultas na rede privada de saúde e conhecimento sobre o ácido fólico. Para diminuir a prevalência de defeitos do tubo neural é importante promoverse o uso do ácido fólico nas mulheres em idade fértil, nas mulheres socioeconomicamente menos favorecidas e disponibilizá-lo na rede pública de saúde.

Introdução - A incidência das fendas lábio – palatinas, é de 1 para cada 700 nascidos vivos na população mundial, trata-se de uma má-formação congênita que pode ocorrer devido a fatores endógenos ou exógenos. Objetivo – Relacionar o índice de massa corporal (IMC) materno com o nascimento de bebês com fendas. Métodos – Trata-se de um estudo tipo caso-controle. Foram incluídas 800 mães com idade entre 18 e 35 anos, que não apresentaram suspeita ou confirmação de diabete mellitus ou gravídica e bebês com peso entre 2.500 e 4.500 gramas nascidos entre a 37ª e a 42ª semana de gestação que não apresentaram nenhum outro tipo de deficiência que não a estudada. Para a coleta dos dados aplicou – se questionários a 400 mães de crianças normais distribuídas em quatro postos de saúde da cidade de Santo André (controles), e 400 mães de bebês com fendas labiais e/ou palatinas que estavam em tratamento no FUNCRAF, que é um centro especializado no tratamento da deformidade estudada, nesta mesma cidade (casos). Resultados - Entre os casos encontrou - se 148 (37,00%) mulheres com IMC acima de 26 e entre os controles foram 132 (33,00%). Com relação ao uso de álcool/drogas observou-se que 82 (20,50%) mulheres entre os casos e 58 (14,50%) no grupo controle eram usuárias. Duzentas e onze (52,70%) mães de bebês com fendas relataram ter histórico de má - formação na família dela ou do pai da criança. Este número foi de 103 (25,70%) no grupo controle. No grupo controle observou-se que 330 (82,50%) eram brancas ou pardas, 43 (10,75%) negras e 27 (6,70%) amarelas, entre os casos foram 294 (73,50%) brancas ou pardas, 65 (16,25%) negras e 40 (10,00%) amarelas. Conclusões - O IMC alto não esteve relacionado ao nascimento de bebês com fendas. Ter sido usuária de álcool/drogas no primeiro trimestre de gestação relacionou-se ao nascimento de crianças com a má-formação. O risco de surgimento desta deformidade foi 3,6 vezes maior quanto existiu histórico de qualquer má-formação familiar; a raça/etnia da mãe também apresentou associação com a presença de fendas, a raça branca é a menos associada seguida pela parda e negra e, finalmente pela amarela que foi a que apresenta maior associação. / Introduction – The incidence of cleft lips and palate is 1 to 700 live births in the world population, it is a congenital malformation that can happens due to endogens or exogenesis factors. Objectives – Association between maternal body mass index (BMI) and the risk of cleft lips or palate. Methods – That is a case – control study. We include 800 mothers between 18 and 35 years old without suspicious of gestational or preexisting diabetes and babies between 2,500 e 4, 000 g was born between the 37 and 42 pregnancy week without other congenital malformation that is not the studied. Some questions were asked for 400 normal children’s mothers from four health centers from Santo André (controls), and 400cleft lips or palate children’s mothers from FUNCRAF that is a health center just to treatment of this malformation, from Santo André (cases). Results – Among the cases we have 148 (37.00%) mothers with BMI above 26 and among the controls it was 132 (33.00%). About alcohol and drugs use we have 82 (20.50%) women among cases and 58 (14.50%) on control group. Two hundred eleven (52.70%) malformations children’s mothers declare to have malformation parental history against 103 (25.70%) on control group. On control group we have 330 (82.50%) white , 43 (10.75%) black and 27 (6.70%) yellow, among the cases 294 (73.50%) white, 65 (16.25%) black and 40 (10.00%) yellow. Conclusions – The increase of BMI is not likely to be a risk factor for clefts, the consumption of alcohol/drugs is a risk factor for this malformation; the risk is 3.6 times highest for clefts when there is malformation history; a racial association with clefts was observed, the white women presents the lowest association and the yellow presents the highest one.

Introdução - A incidência das fendas lábio – palatinas, é de 1 para cada 700 nascidos vivos na população mundial, trata-se de uma má-formação congênita que pode ocorrer devido a fatores endógenos ou exógenos. Objetivo – Relacionar o índice de massa corporal (IMC) materno com o nascimento de bebês com fendas. Métodos – Trata-se de um estudo tipo caso-controle. Foram incluídas 800 mães com idade entre 18 e 35 anos, que não apresentaram suspeita ou confirmação de diabete mellitus ou gravídica e bebês com peso entre 2.500 e 4.500 gramas nascidos entre a 37ª e a 42ª semana de gestação que não apresentaram nenhum outro tipo de deficiência que não a estudada. Para a coleta dos dados aplicou – se questionários a 400 mães de crianças normais distribuídas em quatro postos de saúde da cidade de Santo André (controles), e 400 mães de bebês com fendas labiais e/ou palatinas que estavam em tratamento no FUNCRAF, que é um centro especializado no tratamento da deformidade estudada, nesta mesma cidade (casos). Resultados - Entre os casos encontrou - se 148 (37,00%) mulheres com IMC acima de 26 e entre os controles foram 132 (33,00%). Com relação ao uso de álcool/drogas observou-se que 82 (20,50%) mulheres entre os casos e 58 (14,50%) no grupo controle eram usuárias. Duzentas e onze (52,70%) mães de bebês com fendas relataram ter histórico de má - formação na família dela ou do pai da criança. Este número foi de 103 (25,70%) no grupo controle. No grupo controle observou-se que 330 (82,50%) eram brancas ou pardas, 43 (10,75%) negras e 27 (6,70%) amarelas, entre os casos foram 294 (73,50%) brancas ou pardas, 65 (16,25%) negras e 40 (10,00%) amarelas. Conclusões - O IMC alto não esteve relacionado ao nascimento de bebês com fendas. Ter sido usuária de álcool/drogas no primeiro trimestre de gestação relacionou-se ao nascimento de crianças com a má-formação. O risco de surgimento desta deformidade foi 3,6 vezes maior quanto existiu histórico de qualquer má-formação familiar; a raça/etnia da mãe também apresentou associação com a presença de fendas, a raça branca é a menos associada seguida pela parda e negra e, finalmente pela amarela que foi a que apresenta maior associação. / Introduction – The incidence of cleft lips and palate is 1 to 700 live births in the world population, it is a congenital malformation that can happens due to endogens or exogenesis factors. Objectives – Association between maternal body mass index (BMI) and the risk of cleft lips or palate. Methods – That is a case – control study. We include 800 mothers between 18 and 35 years old without suspicious of gestational or preexisting diabetes and babies between 2,500 e 4, 000 g was born between the 37 and 42 pregnancy week without other congenital malformation that is not the studied. Some questions were asked for 400 normal children’s mothers from four health centers from Santo André (controls), and 400cleft lips or palate children’s mothers from FUNCRAF that is a health center just to treatment of this malformation, from Santo André (cases). Results – Among the cases we have 148 (37.00%) mothers with BMI above 26 and among the controls it was 132 (33.00%). About alcohol and drugs use we have 82 (20.50%) women among cases and 58 (14.50%) on control group. Two hundred eleven (52.70%) malformations children’s mothers declare to have malformation parental history against 103 (25.70%) on control group. On control group we have 330 (82.50%) white , 43 (10.75%) black and 27 (6.70%) yellow, among the cases 294 (73.50%) white, 65 (16.25%) black and 40 (10.00%) yellow. Conclusions – The increase of BMI is not likely to be a risk factor for clefts, the consumption of alcohol/drugs is a risk factor for this malformation; the risk is 3.6 times highest for clefts when there is malformation history; a racial association with clefts was observed, the white women presents the lowest association and the yellow presents the highest one.

No description available.

<p>Developmental toxicology is an important part of preclinical drug toxicology as well as environmental toxicology. Assessing reproductive and developmental toxicity is especially expensive and time demanding, since at least two generations of animals are needed in the tests. In light of this there is a great need for alternative test methods in many areas of developmental toxicity testing.</p><p>The complete set of RNA transcripts in any given organism is called the transcriptome. Proteomics refers to the study of the proteins in a given organism or cell population. The work of this thesis has focused on the use of high throughput screening methods in transcriptomics and proteomics to search for molecular markers of developmental toxicity.</p><p>We have studied the global gene expression effects of the developmentally toxic substance valproic acid (VPA) using microarray technology. Several genes were found that display the same gene expression pattern <i>in vivo</i> using mouse embryos as the pattern seen <i>in vitro</i> using the embryocarcinoma cell line P19. Based on these observations, the gene Gja1 was suggested as one potential molecular marker of VPA induced developmental toxicity and potential marker of histone deacetylase (HDAC) inhibition <i>in vitro</i>. </p><p>Using 2D-DIGE technology, which measures relative protein abundances, the effect of neonatal exposure to the flame retardant PBDE-99 was studied in mouse brain (cortex, hippocampus and striatum) 24 hr after exposure. Differentially expressed proteins in the cortex and the striatum indicate that PBDE-99 may alter neurite outgrowth.</p><p>Finally, we have suggested several improvements in the use of the 2D-DIGE technology. Novel methods for normalizing data were presented, with several advantages compared to existing methods. We have presented a method named DEPPS that makes use of all identified proteins in a dataset to make comprehensive remarks about biological processes affected.</p>

Developmental toxicology is an important part of preclinical drug toxicology as well as environmental toxicology. Assessing reproductive and developmental toxicity is especially expensive and time demanding, since at least two generations of animals are needed in the tests. In light of this there is a great need for alternative test methods in many areas of developmental toxicity testing. The complete set of RNA transcripts in any given organism is called the transcriptome. Proteomics refers to the study of the proteins in a given organism or cell population. The work of this thesis has focused on the use of high throughput screening methods in transcriptomics and proteomics to search for molecular markers of developmental toxicity. We have studied the global gene expression effects of the developmentally toxic substance valproic acid (VPA) using microarray technology. Several genes were found that display the same gene expression pattern in vivo using mouse embryos as the pattern seen in vitro using the embryocarcinoma cell line P19. Based on these observations, the gene Gja1 was suggested as one potential molecular marker of VPA induced developmental toxicity and potential marker of histone deacetylase (HDAC) inhibition in vitro. Using 2D-DIGE technology, which measures relative protein abundances, the effect of neonatal exposure to the flame retardant PBDE-99 was studied in mouse brain (cortex, hippocampus and striatum) 24 hr after exposure. Differentially expressed proteins in the cortex and the striatum indicate that PBDE-99 may alter neurite outgrowth. Finally, we have suggested several improvements in the use of the 2D-DIGE technology. Novel methods for normalizing data were presented, with several advantages compared to existing methods. We have presented a method named DEPPS that makes use of all identified proteins in a dataset to make comprehensive remarks about biological processes affected.