O timo é o órgão linfóide primário responsável pelo estabelecimento inicial de um repertório funcional de células T. A via de sinalização Notch é essencial para o desenvolvimento de células T a partir de células-tronco hematopoiéticas, e a distribuição de seus receptores e ligantes no timo humano ainda é desconhecida. A expressão de AIRE é crucial para a seleção de um repertório de receptores de linfócitos T (TCR) sem autorreatividade. Neste estudo, analisamos o padrão de expressão de AIRE e a distribuição de Notch em timos pacientes com cardiopatias congênitas, parte dos quais com síndrome de Down. Descrevemos a localização intratímica e os tipos celulares capazes de expressar os diferentes receptores e ligantes Notch. A expressão de AIRE em células epiteliais medulares foi significantemente reduzida em timos de crianças com síndrome de Down, deficiência esta que pode explicar a alta incidência de doenças autoimunes nesta cromossomopatia. / The thymus is a primary lymphoid organ which is essential for the initial establishment of a functional repertoire of T cells. Notch signaling is crucial for T-cell lineage development from hematopoietic stem cells; however, distribution of Notch ligands and receptors in human thymus is still unknown. AIRE is crucial for the selection of a T-cell-receptor (TCR) repertoire purged of self-reactive specificities. In this study, we analyzed the expression patterns of AIRE and Notch in human thymuses from children with congenital cardiopathies that undergo heart surgery, part of whom with Down syndrome. We described the intra-thymic localization and the cell types that express Notch receptors and ligands. AIRE expression in medullary epithelial cells is significantly decreased in Down syndrome patients. This deficiency could explain higher incidence of autoimmune disease in Down syndrome.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2016-07-26T15:07:30Z No. of bitstreams: 1 alexandremirandapereira_dissert.pdf: 2691507 bytes, checksum: 59310c2c32ff23d4dc858c5d7bf68dd8 (MD5) / Made available in DSpace on 2016-07-26T15:07:30Z (GMT). No. of bitstreams: 1 alexandremirandapereira_dissert.pdf: 2691507 bytes, checksum: 59310c2c32ff23d4dc858c5d7bf68dd8 (MD5) Previous issue date: 2013-03-25 / The Tempormandibular joint (TMJ) is considered to be the most complex of the human body. It is classified as synovial, and then presents capsule and articular disc, membrane, synovial fluid and ligaments. Particularly, TMJ is a form of connection between the mandibular condyle (face) and temporal bone (skull), which generates a mechanically stable joint component. There are currently few studies concerning the study of TMJ in patients with Down syndrome (DS) (trisomy 21). The aim of this study is to morphologically analyze the mandibular condyle by means of Cone Beam Computed Tomography (CBCT) in young individuals with or without SD. After ethical approval, 23 patients were allocated (male or female and aged between 18 and 20 years), 10 patients with SD (study group) and 13 young people without such Syndrome. Morphological analysis (size and shape in three-dimensional approach) of the mandibular condyle was bilaterally determined by i-CAT Cone Beam Tomography (cone beam ®) imaging detector. The classification of the shape and size of the mandibular condyle was obtained by visualizations of coronal and sagittal planes, as well as by mandibular points. The statistical analyzes included determining the frequency of occurrence for the parameters considered above, with the use of Fisher's exact test, as well as estimating the average continuous quantitative variables by Student's t test. The average size of the mandibular condyle in the control group, left and right sides was 9.71 mm and 9.58 mm in sagittal analysis, while in the Group with Down syndrome was 10.15 mm and 9.97 mm. In coronal analysis the average size of the mandibular condyle, left and right sides, in the control group was 15.59 mm and 15.49 mm, while in the group with Down syndrome was 14.53 mm and 15.12 mm. Cone Beam Computed Tomography (CBCT) is a excellence imaging method employing reduced amount of radiation in a single measurement, low cost and versatility for image reconstruction. In the morphological analysis of the mandibular condyle of young people with or without Down syndrome detected by Cone Beam CT by sagittal and coronal visualizations, there were no significant changes of shape (round and straight) nor size (distance between the extreme anteroposterior and mediolateral mandibular points) comparing both groups. It is suggested that the continued growth potential of the mandibular condyle by temporomandibular joint cartilage remodeling can justify the results. Using tomographic additional incidences associated with a larger sample size could definitively corroborate the findings of this study. / A Articulação Tempormandibular (ATM) é considerada a mais complexa do corpo humano. Classificada como sinovial, e assim apresenta cápsula e disco articulares, membrana e líquido sinoviais e ligamentos. Particularmente, ATM é uma forma de ligação entre a cabeça da mandíbula (face) e o osso temporal (crânio), o que gera um componente articular mecanicamente estável. Há, no momento, escassos estudos referentes ao estudo da ATM em pacientes com Síndrome de Down (SD) trissomia do 21. O objetivo do presente estudo é analisar morfologicamente a cabeça da mandíbula por meio de Tomografia Computadorizada (TC) de Feixe Cônico em indivíduos jovens, portadores ou não, de SD. Após anuência ética, foram alocados 23 jovens (sexo masculino ou feminino e idade entre 18 e 20 anos), 10 portadores de SD (grupo de estudo) e 13 jovens sem a referida Síndrome. Avaliação morfológica (tamanho e forma em abordagem tridimensional) da cabeça da mandíbula foi determinada bilateralmente por tomógrafo i-CAT do tipo feixe cônico (cone beam®) com detector de imagem. A classificação da forma e do tamanho da cabeça da mandíbula foi obtida pelas visibilizações nos planos coronal e sagital, bem como pela mensuração da distância entre os pontos mandibulares extremos anteroposterior e médio-lateral. Análise estatística compreendeu determinação das frequências de ocorrência para os parâmetros acima considerados, com o emprego do Teste Exato de Fisher, bem como a estimativa da média das variáveis quantitativas contínuas pelo Teste t de Student. A média do tamanho da cabeça da mandíbula do Grupo Controle, do lado direito e esquerdo, em análise sagital foi de 9,71mm e 9,58mm, sendo que no Grupo com Síndrome de Down foi de 10,15mm e 9,97mm. Em análise coronal a média do tamanho da cabeça da mandíbula, lado direito e esquerdo, no Grupo Controle foi de 15,59 mm e 15,49 mm, enquanto que no Grupo com Síndrome de Down foi de 14,53 mm e 15,12 mm. Tomografia Computadorizada (TC) de Feixe Cônico é método de imagem de excelência que emprega reduzida quantidade de radiação em tomada única, a baixo custo e com versatilidade para reconstrução de imagens. Na análise morfológica da cabeça da mandíbula de jovens com ou sem Síndrome de Down por TC de Feixe Cônico por visibilização sagital e coronal, não foram encontradas alterações significantes de formato (arredondado e reto) e nem de tamanho (distância entre os pontos mandibulares extremos anteroposterior e médio-lateral) na comparação de ambos os grupos. Sugere-se que o contínuo potencial de crescimento da cabeça da mandíbula por remodelação da cartilagem da Articulação Temporomandibular possa justificar os resultados encontrados. Uso de incidências tomográficas adicionais, associadas a um maior tamanho amostral, poderiam corroborar em definitivo os achados do presente estudo.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2018-02-15T12:46:24Z No. of bitstreams: 1 marianafernandadossantos_dissert.pdf: 1735247 bytes, checksum: 1bde0ab992e930a0190504964f47726e (MD5) / Made available in DSpace on 2018-02-15T12:46:25Z (GMT). No. of bitstreams: 1 marianafernandadossantos_dissert.pdf: 1735247 bytes, checksum: 1bde0ab992e930a0190504964f47726e (MD5) Previous issue date: 2016-12-01 / Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP / Introduction: Congenital heart defects (CHD) are present in approximately 40 to 60% of individuals with Down syndrome (DS). It is the leading cause of death in the first years of life in individuals with the syndrome. Polymorphisms in maternal and fetal genes encoding enzymes involved in folate metabolism have been associated with the development of congenital heart defects. Objectives: To assess if the presence of polymorphism (MTHFR rs4846048, MTHFR rs4846049, hsa-mir-149 rs2292832, MTHFR C677T, MTHFR A1298C, MTHFR T1317C, MTR A2756G, MTRR A66G, SLC19A1 A80G, TC2 A67G, TC2 C776G, CßS 844ins68, CßS T833C, MTHFD1 G1958A, BHMT G742A, DHFR del 19 pb, and SHMT C1420T) in individuals with DS is associated with the occurrence of CHD in these individuals. We also evaluated the association between maternal genetic polymorphisms MTHFR rs4846048, MTHFR rs4846049 and hsa-mir-149 rs2292832, and the presence of CHD in offspring with DS. Methods: This study included 139 individuals (80 individuals with DS and CHD, and 59 control subjects with DS without congenital heart disease). Molecular analysis of MTHFR rs4846048, MTHFR rs4846049 and hsa-mir-149 rs2292832 was carried out by real time polymerase chain reaction allelic discrimination. Genotyping data of MTHFR C677T, MTHFR A1298C, MTHFR T1317C, MTR A2756G, MTRR A66G, SLC19A1 A80G, TC2 A67G, TC2 C776G, CßS 844ins68, CßS T833C, MTHFD1 G1958A, BHMT G742A, DHFR del 19 pb, and SHMT C1420T were obtained from database of previous studies of the research group and also used to assess the risk for the occurrence of CHD in this study. Multiple logistic regression analyzes were performed to assess the risk of CHD in the presence of 17 polymorphisms in dominant and recessive genetic models. The median number of mutant alleles between groups was assessed by the Mann-Whitney test. Genotypic combination analysis was performed for the polymorphisms MTHFR rs4846048, MTHFR rs4846049, and hsa-mir-149 rs2292832, using Fisher's exact test, dominant model. Results: Multiple logistic regression analysis involving individuals with DS showed no association between 17 polymorphisms and the risk for CHD. The median number of polymorphic alleles did not differ among individuals with DS with and without CHD. On the other hand, the maternal genotypes hsa-mir-149 rs2292832 CT or TT were associated with reduced risk for isolated heart disease in the offspring (OR = 0,31; 95% CI = 0,13 to 0,72; P = 0,0063). The analysis of genotypic combinations of MTHFR rs4846048, MTHFR rs4846049, and hsa-mir-149 rs2292832 in individuals with DS, and their mothers showed no association between the different combinations and the risk for congenital heart disease. Conclusions: There is no evidence of association between the polymorphisms analyzed in individuals with DS and the occurrence of CHD. However, a lower risk of isolated congenital heart disease for individuals with DS is observed in the presence of maternal genotypes hsa-mir-149 rs2292832 CT or TT. / Introdução: Defeitos cardíacos congênitos (DCC) estão presentes em aproximadamente 40 a 60% dos indivíduos com a síndrome de Down (SD) e representam a principal causa de morte nos primeiros anos de vida em indivíduos com a síndrome. Polimorfismos em genes maternos e fetais, que codificam enzimas envolvidas no metabolismo do folato, têm sido associados com o desenvolvimento de cardiopatias congênitas. Objetivos: Avaliar se a presença dos polimorfismos MTHFR rs4846048, MTHFR rs4846049, hsa-mir-149 rs2292832, MTHFR C677T, MTHFR A1298C, MTHFR T1317C, MTR A2756G, MTRR A66G, SLC19A1 A80G, TC2 A67G, TC2 C776G, CßS 844ins68, CßS T833C, MTHFD1 G1958A, BHMT G742A, DHFR del 19 pb, SHMT C1420T em indivíduos com SD está associada com a ocorrência de DCC nesses indivíduos. Também foi avaliada a associação entre os polimorfismos genéticos maternos MTHFR rs4846048, MTHFR rs4846049 e hsa-mir-149 rs2292832 e a presença de DCC na prole com SD. Casuística e Método: Este estudo incluiu 139 indivíduos (80 indivíduos com SD e DCC e 59 indivíduos controles com SD, sem cardiopatia congênita). A análise molecular dos polimorfismos MTHFR rs4846048, MTHFR rs4846049 e hsa-mir-149 rs2292832 foi realizada pelo método discriminação alélica por meio de reação em cadeia da polimerase em tempo real. Os dados da genotipagem dos polimorfismos MTHFR C677T, MTHFR A1298C, MTHFR T1317C, MTR A2756G, MTRR A66G, SLC19A1 A80G, TC2 A67G, TC2 C776G, CßS 844ins68, CßS T833C, MTHFD1 G1958A, BHMT G742A, DHFR del 19 pb, SHMT C1420T foram obtidos de banco de dados de trabalhos previamente publicados pelo grupo de pesquisa e utilizados para avaliar o risco para a ocorrência de DCC no presente estudo. Análises de regressão logística múltipla foram realizadas para avaliar o risco de DCC na presença dos 17 polimorfismos nos modelos genéticos dominante e recessivo. A mediana do número de alelos mutantes entre os grupos foi avaliada pelo teste de Mann-Whitney. Análise de combinação genotípica foi realizada para os polimorfismos MTHFR rs4846048, MTHFR rs4846049 e hsa-mir-149 rs2292832, utilizando o teste exato de Fisher, no modelo dominante. Resultados: As análises de regressão logística múltipla, envolvendo os indivíduos com SD, não evidenciaram associação entre os 17 polimorfismos e o risco para DCC. A mediana do número de alelos polimórficos também não diferiu entre os indivíduos com SD com e sem DCC. Por outro lado, os genótipos maternos hsa-mir-149 rs2292832 CT ou TT foram associados ao risco reduzido para cardiopatia isolada na prole com SD (OR = 0,31; IC 95% = 0,13-0,72; P = 0,0063). A análise das combinações genotípicas dos polimorfismos MTHFR rs4846048, MTHFR rs4846049 e hsa-mir-149 rs2292832, nos indivíduos com SD e nas suas mães, não mostrou associação entre as diferentes combinações e o risco para cardiopatia congênita. Conclusões: Na casuística avaliada não há evidências de associação entre os polimorfismos analisados em indivíduos com SD e a ocorrência de DCC; entretanto um menor risco de cardiopatia congênita isolada para os indivíduos com SD é observado na presença dos genótipos maternos hsa-mir-149 rs2292832 CT ou TT.

Resumo: Por meio de exames clínicos e análises laboratoriais, o presente trabalho avaliou a prevalência de cárie dentária e a relação entre os fatores presença de cárie dental, biofilme bacteriano visível e contagem de Streptococcus grupo mutans em crianças com idade entre 12 e 48 meses, sendo 26 portadoras (grupo teste) e 142 não portadoras da síndrome de Down (grupo controle). A prevalência de cárie dentária no grupo teste foi de 15,38% e no grupo controle de 31,69%. No grupo controle houve aumento estatisticamente significativo do índice ceo-d e dos níveis de Streptococcus grupo mutans a partir dos 36 meses de idade. Nos dois grupos, com o aumento da idade, aumentou o número de crianças colonizadas por Streptococcus grupo mutans. Foi observada dependência estatisticamente significativa entre a presença de lesão cárie dentária e altos níveis de Streptococcus do grupo mutans tanto no grupo teste quanto no grupo controle. Houve também correlação positiva estatisticamente significativa entre a presença de lesão de cárie dentária e a presença de biofilme bacteriano visível, e entre a presença de biofilme bacteriano visível e altos níveis de Streptococcus do grupo mutans apenas no grupo controle. / Abstract: This study evaluated the dental caries prevalence and the relationship among dental caries, visible bacterial biofilm and mutans streptococci counts in children with Down syndrome (test-group=26) and in health children (control-group=142), aged from 12 to 48 months. The caries prevalence was 15.38% in the test group and 31.69% in the control-group. The dmf-t index and the mutans streptococci levels had a significant statistic increase after 36 months of age in the control group. The number of children who harbored mutans streptococci had a significant increased with age in all children (test and control groups). In both groups dental caries and high levels of mutans streptococci presented a strong positive correlation. High positive correlation was also observed between visible bacterial biofilm and dental caries lesions and between visible bacterial biofilm and high levels of mutans streptococci in the control-group, but not in the test-group. Keywords: Down syndrome, dental caries, mutans streptococci, biofilm. / Orientador: Ângela Cristina Cilense Zuanon / Coorientador: Denise Madalena Palomari Spolidorio / Banca: Fábio César Braga de Abreu e Lima / Banca: Cássia Cilene Dezan Garbelini / Mestre

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-06-11T15:51:18Z No. of bitstreams: 1 Dissertação - Alessandra Valle Salino.pdf: 2481289 bytes, checksum: 21867cc6aefc275133666885d85903bf (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-11T20:40:14Z (GMT) No. of bitstreams: 1 Dissertação - Alessandra Valle Salino.pdf: 2481289 bytes, checksum: 21867cc6aefc275133666885d85903bf (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-11T20:41:20Z (GMT) No. of bitstreams: 1 Dissertação - Alessandra Valle Salino.pdf: 2481289 bytes, checksum: 21867cc6aefc275133666885d85903bf (MD5) / Made available in DSpace on 2015-06-11T20:41:20Z (GMT). No. of bitstreams: 1 Dissertação - Alessandra Valle Salino.pdf: 2481289 bytes, checksum: 21867cc6aefc275133666885d85903bf (MD5) Previous issue date: 2014-02-28 / Não Informada / Several studies with different pathologies or injuries have linked oral health to quality of life by emphasizing the importance of studying the subjective aspects related to oral diseases. However, there has been few research on some conditions, including Down syndrome. The aim of this study is to describe the health-related quality of life and impact of oral health on quality of life of individuals with Down syndrome, in Manaus, Amazonas. The quality of life and quality of life related to oral health factors associated, were evaluated by the instruments PedsQL 4.0 and OHIP-14 were also identified . The questionnaires were administered to the mothers of the studied subjects. Dependent variables included caries experience (DMFT), treatment needs and periodontal disease (CPI and PIP). This study is descriptive, analytical and cross-sectional. The study population was comprised of individuals from 06 years old with Down syndrome who were registered in institutions APAE-Manaus, Amazonas Pestalozzi Society, Municipal Complex Special Education "André Vidal de Araújo" and Amazonas State University - UEA. The prevalence of dental caries was 62,1% and mean DMFT was 6,25. The presence of bleeding and dental calculus, in at least 1 site, was found in 69,01% and 66,20%, respectively. The prevalence of shallow pocket and deep was 45,07% and 18,31%. The data suggests oral health status in terms of caries and periodontal disease, although present in most of the study population, showed few impact on quality of life. Considering the prevalence of caries and periodontal disease and oral morbidity found, it is necessary, monitoring the oral health of individuals with Down syndrome. / Diversos estudos com diferentes patologias ou agravos têm relacionado à saúde bucal com a qualidade de vida, apontando a importância de estudos voltados para os aspectos subjetivos afetados pelas doenças bucais. Entretanto, ainda há escassez de estudos em algumas condições, incluindo a síndrome de Down. O objetivo deste estudo foi descrever a qualidade de vida relacionada à saúde e o impacto da saúde bucal na qualidade de vida de indivíduos com síndrome de Down, em Manaus, Amazonas. Foram identificados, ainda, fatores associados à qualidade de vida e à qualidade de vida relacionada à saúde bucal, avaliados através dos instrumentos PedsQL 4.0 e o OHIP-14. Os questionários foram aplicados às mães dos sujeitos do estudo. As variáveis independentes incluíram experiência de cárie (CPO-D), necessidade de tratamento e doença periodontal (CPI e PIP). O estudo foi do tipo descritivo, analítico e transversal. A população do estudo foi composta por indivíduos, a partir de 06 anos de idade, com síndrome de Down, cadastrados nas instituições APAE - Manaus, Sociedade Pestalozzi do Amazonas, Complexo Municipal de Educação Especial “André Vidal de Araújo” e Universidade do Estado do Amazonas - UEA. A prevalência de cárie dentária foi 62,1% e o CPO-D médio foi 6,25. A presença de sangramento e cálculo dentário, em pelo menos 1 sítio, foi encontrada em 69,01% e 66,20%, respectivamente. A prevalência de bolsa rasa e profunda foi 45,07% e 18,31%. Os dados sugerem que a condição de saúde bucal em termos de cárie e doença periodontal, embora, presente na maioria da população do estudo mostrou pouco impacto na qualidade de vida. Diante da prevalência de cárie e doença periodontal encontrada e morbidade bucal referida, se faz necessário um monitoramento da saúde bucal dos indivíduos com síndrome de Down.

Os pacientes com Síndrome de Down (SD) possuem grande incidência de doença periodontal (DP), caracterizada por um curso precoce e com maior severidade. O estudo de metaboloma pode contribuir para o entendimento deste curso da doença, identificando possíveis metabólitos como biomarcadores nestes indivíduos. Para entender o perfil metabolômico dos indivíduos com síndrome de Down e a sua relação com a doença periodontal, realizamos a identificação de metabólitos salivares de adolescentes e adultos jovens, entre 12 e 21 anos, ambos os gêneros. Foram coletados dados sobre o estado geral de saúde e realizados exames clínicos bucais, como índice de higiene oral simplificado, sangramento e profundidade de sondagem. Para a análise do metaboloma foi coletada amostra de saliva não estimulada, analisadas por meio de cromatografia gasosa acoplada á espectrometria de massas. Saliva e fluido crevicular gengival também foram coletados para identificação microbiana através do MALDI-TOF. Os dados encontrados foram submetidos a análise estátisca por meio da Análise dos Componentes Principais (PCA) e quantificação relativa dos metabólitos foi avaliada por testes não paramétricos, Mann-Whitney e Kruskal-Wallis. Foi possível observar através dos modelos de PCA separação dos indivíduos com SD e controles, independente da doença periodontal. A quantificação relativa revelou maiores níveis de glicina, lprolina, l-leucina, l-serina, ácido palmítico, ácido pentanóico, ácido tetradecanóico, tirosina e l-fenilalanina nos grupos SD quando comparados aos controles. Controles com DP também apresentaram níveis elevados de glicina, l-alanina, l-serina e manopiranose quando comparados com controles saudáveis. A microbiota de indivíduos com SD apresentous diferenças siginificantes em relação aos individuos controles, principalmente para Rothia dentocariosa, Staphylococcus epidermidis, Tannerella forsythia quando avaliado a saliva e A. Actinomycetemcomitans, Micrococcus luteus, Rothia aeria, Treponema denticola no fluido crevicular gengival. Em conclusão, o perfil metabolômico impresso nos indivíduos com SD difere significativamente dos indivíduos controles, independente da doença periodontal. Entretanto, os metabólitos que diferenciam indivíduos controles com e sem DP, apresentam-se elevados em todos indivíduos com SD, promovendo novos \"insights\" para o perfil metabólico relacionado a DP na SD. / Down Syndrome (DS) patients have a high incidence of periodontal disease (PD), characterized by an early course and greater severity. The metabolome study may contribute to the understanding of the disease course, identifying possible metabolites as biomarkers in these individuals. To understand the metabolomic profile of the DS and their relationship with PD, we conducted the identification of salivary metabolites of adolescents and young adults between 12 and 21 years, both genders. Data were collected on general health and was performed oral clinical examination, as the IHOS, bleeding index and probing depth. For metabolome analysis was collected unstimulated saliva sample, analyzed by gas chromatography coupled to mass spectrometry. Saliva and gingival crevicular fluid were also collected for microbial identification by MALDI-TOF. Data were submitted to analysis-statistic by PCA and relative quantification of metabolites was evaluated by Mann-Whitney and Kruskal-Wallis tests. It can be observed through the PCA models separation of DS groups and controls groups, regardless of periodontal disease. Relative quantification showed higher levels of glycine, L-proline, L-leucine, L-serine, palmitic acid, pentanoic acid, tetradecanoic acid, tyrosine and L-phenylalanine in the SD groups when compared to controls groups. Controls with PD also showed high levels of glycine, L-alanine, L-serine and mannopyranose compared with healthy controls. The microbiota of individuals with DS groups show significant differences compared to control groups, especially for Rothia dentocariosa, Staphylococcus epidermidis, Tannerella forsythia when evaluated saliva and A. actinomycetemcomitans, Micrococcus luteus, Rothia aeria, Treponema denticola in gingival crevicular fluid. In conclusion, the printed metabolomic profile in individuals with Down syndrome differs significantly from control subjects, regardless of periodontal disease. However, the metabolites that distinguish controls group with and without PD, show up high in all DS individuals, promoting new \"insights\" to the metabolic profile related to PD in DS.

Chan, Ka Ying. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 133-150). / Abstract also in Chinese. / ABSTRACT --- p.i / 摘要 --- p.iii / ACKNOWLEDGEMENTS --- p.iv / CONTRIBUTORS --- p.vi / TABLE OF CONTENTS --- p.vii / LIST OF TABLES --- p.x / LIST OF FIGURES --- p.xi / LIST OF ABBREVIATIONS --- p.xiii / Chapter SECTION I: --- BACKGROUND --- p.1 / Chapter CHAPTER 1: --- PRENATAL DIAGNOSIS OF FETAL TRISOMY 21 --- p.2 / Chapter 1.1 --- Down syndrome --- p.2 / Chapter 1.2 --- Current methods of prenatal diagnosis of fetal trisomy 21 --- p.3 / Chapter 1.2.1 --- Non-invasive procedures --- p.3 / Chapter 1.2.2 --- Invasive procedures --- p.5 / Chapter 1.3 --- Alternative methods for the prenatal diagnosis of fetal trisomy 21 --- p.7 / Chapter CHAPTER 2: --- CELL-FREE FETAL NUCLEIC ACIDS IN MATERNAL PLASMA --- p.13 / Chapter 2.1 --- Circulating fetal cells --- p.15 / Chapter 2.2 --- Circulating cell-free fetal nucleic acids --- p.15 / Chapter 2.3 --- Diagnostic applications of cell-free fetal nucleic acids in maternal plasma --- p.17 / Chapter 2.4 --- Digital relative chromosome dosage approach --- p.20 / Chapter 2.5 --- Validation of digital RCD approach on artificial DNA mixtures --- p.22 / Chapter SECTION II --- : MATERIALS AND METHODS --- p.25 / Chapter CHAPTER 3: --- QUANTITATIVE ANALYSIS OF NUCLEIC ACIDS --- p.26 / Chapter 3.1 --- Subject recruitment and sample collection --- p.26 / Chapter 3.2 --- Sample processing --- p.26 / Chapter 3.3 --- Nucleic acid extraction --- p.27 / Chapter 3.3.1 --- Extraction of DNA from placental tissues --- p.27 / Chapter 3.3.2 --- Extraction of DNA from maternal blood cells --- p.27 / Chapter 3.4 --- Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) --- p.28 / Chapter 3.5 --- Paralogous sequence assays optimisation workflow --- p.31 / Chapter 3.5.1 --- Monoplex paralogous sequence assays --- p.31 / Chapter 3.5.2 --- Multiplex paralogous sequence assay --- p.38 / Chapter 3.6 --- Digital PCR --- p.42 / Chapter 3.6.1 --- Principle --- p.42 / Chapter 3.6.2 --- Digital multiplex paralogous sequence assay --- p.42 / Chapter 3.7 --- Statistical analysis --- p.46 / Chapter 3.7.1 --- Disease classification of samples --- p.46 / Chapter 3.7.2 --- Poisson distribution --- p.46 / Chapter 3.7.3 --- Data analysis --- p.48 / Chapter 3.7.4 --- Sequential probability ratio test (SPRT) analysis --- p.49 / Chapter SECTION III: --- ASSAY DEVELOPMENT --- p.53 / Chapter CHAPTER 4: --- TESTING OF ASSAY SPECIFICITY WITH CORIELL CELL LINES --- p.54 / Chapter 4.1 --- Coriell cell lines --- p.54 / Chapter 4.2 --- Specificity of initial PCR primers --- p.56 / Chapter 4.2.1 --- Principle --- p.56 / Chapter 4.2.2 --- Materials and methods --- p.56 / Chapter 4.2.3 --- Results --- p.60 / Chapter 4.2.4 --- Conclusion --- p.63 / Chapter 4.3 --- Specificity of the iPLEX® Gold extension primers --- p.63 / Chapter 4.3.1 --- Principle --- p.63 / Chapter 4.3.2 --- Materials and methods --- p.64 / Chapter 4.3.3 --- Results --- p.65 / Chapter 4.4 --- Further analysis on the specificity of PV2107a initial PCR primers --- p.67 / Chapter 4.5 --- Conclusion --- p.71 / Chapter CHAPTER 5: --- ASSAY OPTIMISATION --- p.72 / Chapter 5.1 --- Introduction --- p.72 / Chapter 5.2 --- Optimisation of initial PCRs with AmpliTaq Gold® DNA polymerase followed by homogeneous MassEXTEN´DёØ (hME) assays (Sequenom) --- p.72 / Chapter 5.2.1 --- Optimisation of initial PCR reactions --- p.72 / Chapter 5.2.2 --- Principle of homogeneous MassEXTEN´DёØ assays (Sequenom)… --- p.75 / Chapter 5.2.3 --- Homogeneous MassEXTEN´DёØ assays (Sequenom) on euploid and T21 samples --- p.76 / Chapter 5.3 --- Assay selection by iPLEX® Gold single base primer extension reactions (Sequenom) --- p.82 / Chapter 5.4 --- Optimisation of multiplex PCR with AmpliTaq Gold® DNA polymerase --- p.88 / Chapter 5.5 --- Optimisation of multiplex iPLEX® Gold single base primer extension reaction --- p.93 / Chapter 5.6 --- Single molecule detection test for the multiplex paralogous sequence assays … --- p.103 / Chapter SECTION IV: --- ANALYSIS OF CLINICAL SAMPLES --- p.107 / Chapter CHAPTER 6: --- DISEASE CLASSIFICATION OF EUPLOID AND TRISOMY SAMPLES WITH MULTIPLEX PARALOGOUS SEQUENCE ASSAY --- p.108 / Chapter 6.1 --- Introduction --- p.108 / Chapter 6.2 --- Materials and methods --- p.109 / Chapter 6.2.1 --- Sample collection --- p.109 / Chapter 6.2.2 --- Experimental design --- p.110 / Chapter 6.3 --- Results --- p.111 / Chapter 6.4 --- Discussion --- p.114 / Chapter SECTION V: --- CONCLUDING REMARKS --- p.122 / Chapter CHAPTER 7: --- CONCLUSION AND FUTURE PERSPECTIVES --- p.123 / Chapter 7.1 --- Conclusion --- p.123 / Chapter 7.2 --- Future perspectives --- p.124 / Appendix 1 --- p.126 / Appendix II --- p.127 / REFERENCE --- p.133

Parenting self-efficacy is one factor identified as relevant to parent distress and child therapy outcomes. Theories for parenting self-efficacy suggest parents of children with autism spectrum disorders (ASD) may be at risk for lower parenting self-efficacy than other parents. Parents who have low parenting self-efficacy may then have higher risk for poor treatment outcomes. Previous researchers found inconsistent results related to parenting self-efficacy rates for parents of children with ASD. They suggested the results were due to sample sizes, measurement insensitivitiy, comparison groups, and the limited range of children's ages (Fields, 2006; Meirsschaut, Roeyers, and Warreyn, 2010; Palafox, 2004; and Rutgers et al., 2007). In the current study, the researchers invited 598 parents to complete a series of questionnaires. Participants included the parents of children with ASD (n = 57), Down syndrome (n = 24), ASD and Down syndrome (n = 41), emotional and behavioral disorders (n = 287), and no identified diagnoses (n = 189). The parents who participated were 90.2% female and 84.9% Caucasian. Participants from the ASD, ASD with Down syndrome, and Down syndrome groups lived in higher income households (75.2% above $30,000 annually) than those in the emotional and behavioral disorder group (94.1% below $30,000 annually). The questionnaires asked parents to rate themselves regarding parenting self-efficacy, parent distress, parenting skills, social support, and answered demographic questions. Parents from the diagnostic groups also rated their child's behavior and symptom severity. Parents from the ASD, Down syndrome, and ASD with Down syndrome groups answered additional questions found to be relevant in Fields, 2007 (e.g. age of symptom onset, number of siblings, and parent's age). Parents of children with ASD were found to have the lowest rates of parenting self-efficacy across the five groups. ANOVA rejected the null hypothesis that the groups would be the same (F = 8.24, df = 4, 595, p < .01, adjusted R² = .05). The effect size for the relationship between diagnosis and parenting self-efficacy was small to moderate, accounting for 5% of the variance of parenting self-efficacy scores. Pairwise comparisons between groups found parents of children with ASD to have significantly lower parenting self-efficacy than the Down syndrome (mean difference = -3.32, se = .81, 95% CI = -5.86, -.78), and community groups (mean difference = -2.89, se = .58, 95% CI = -4.47 to -1.31). Parents from the community group were also found to have higher parenting self-efficacy than the parents of children with emotional and behavioral disorders (mean difference = 1.43, se = .37, 95% CI = 1.31, 4.47). Parenting self-efficacy was also related to parent distress, social support, parenting skills, and child's age. Parenting self-efficacy may warrant monitoring in the treatment of ASD and may be an important point of intervention in therapy.

Age-related neurologic disease is a significant and growing burden on our society. Although the largest share of research effort has typically been devoted to the common neurodegenerative illnesses (such as Alzheimer’s disease, or AD), the reality is that nearly all cases of neurodegenerative disease possess elements of mixed pathology. Vascular contributions to cognitive impairment and dementia (VCID) is a complex form of dementia, combining aspects of vascular disease and other forms of dementia, such as Alzheimer’s disease. This pathology is heterogeneous and can include cerebral amyloid angiopathy (CAA), hemorrhages, white matter infarcts, and changes to the neurovascular unit. Given the heterogeneous nature of VCID, we hypothesized that we could further elucidate mechanisms that drive dementia in VCID by examining pathology in mouse models and use this data to guide the study of human autopsy cases. Using a mouse model of VCID, we identified NHE1, a sodium hydrogen exchanger that was upregulated in these mice, as a possible candidate for a factor involved in cerebrovascular disease in humans. We saw a significant age effect of NHE1 in cases with Down syndrome (DS), leading us to further examine cerebrovascular pathology in individuals with DS. People with DS are at a high risk of developing cognitive impairment and dementia after the age of 50. In fact, virtually all adults with DS develop the neuropathology for an AD (beta-amyloid (Aß) senile plaques and tau neurofibrillary tangles) diagnosis by the age of 40 due to a triplication of chromosome 21. We found that these individuals develop CAA and microhemorrhages as a function of age, and that these rates are as severe as sporadic AD, despite an age difference of ~30 years. We also found that individuals with DS have different microglial morphologies than controls or individuals with AD. This data indicates that people with DS develop significant cerebrovascular and AD pathology, indicative of VCID. Overall, we found that mixed pathologies, specifically VCID, is an important contributor to the development of dementia and should be studied further to better understand how this pathology drives cognitive impairment. Further, it is clear that mouse models map imperfectly onto complex human diseases, and that significant work remains to be done towards achieving an adequate model of VCID.

This thesis explores the meaning of wellbeing for children with a disability in New Zealand, an area of social policy that has been largely unexamined. Focusing on the school environment, three questions are addressed: What does wellbeing mean for children with a disability? What factors influence it? Are current policy frameworks which address child wellbeing relevant to the wellbeing of children with a disability? The research involved qualitative data collection from nine purposively selected participants: children with a disability, their parents and key informants involved in service provision and policy development. A critical review of international and national literature on definitions of wellbeing and disability, and on existing data sources, is followed by a socio-demographic profile of children with a disability in New Zealand. Qualitative findings are interpreted in relation to current New Zealand social policy initiatives and frameworks - New Zealand's Agenda for Children, the Whole Child Approach and the Key Settings Model - as well as the theoretical perspectives of social solidarity, wellbeing, the ecological theory of human development and discourses of disability. Findings indicate that the concept of wellbeing as applied to all New Zealand children is also relevant to children with a disability. The difference however, lies in the factors which ultimately influence whether the various dimensions of wellbeing will actually be experienced by children with a disability. For these children, communication as a dimension of wellbeing for example, is influenced by language skill acquisition, which in turn depends upon allocation of appropriate and adequate resourcing of the child's learning environment. The conclusion drawn is that policy frameworks, principles and social indicators addressing child wellbeing, are inconsistently applied with regard to children with a disability. New Zealand's Agenda for Children which promotes an ecological approach to child wellbeing would benefit from further adaptation to reflect the needs of this specific child population. The notion of wellbeing for children with a disability needs further development for the purpose of knowledge building, and to ensure clearer articulation between processes of policy development, service provision, and resource allocation.