Thesis (MSc (Genetics))--University of Stellenbosch, 2009. / Abruptio placentae is the premature separation of the normally implanted placenta from the uterine wall, resulting in haemorrhage before delivery of the fetus. This has serious maternal and neonatal implications, and is one of the leading causes of perinatal and maternal mortality and morbidity in South Africa. Placental vasculopathies, such as abruptio placentae, are believed to result from faults occurring in early placental development. Placental protein 13 (PP13) is a member of the pregnancy-related protein family, and is believed to function in a number of important physiological processes such as trophoblast invasion, placentation and implantation. The aim of this study was to investigate whether DNA sequence variants in the LGALS13 gene (encoding PP13), underlie and/or confer susceptibility to abruptio placentae. The gene was screened and genotyped in a cohort of patients whose pregnancies were complicated by abruptio placentae, as well as an ethnically matched control cohort. Statistical and in silico analyses were performed in order to identify potential susceptibility factors in this South African cohort and to predict whether the identified variants may impact on gene expression or the structure and function of PP13. In addition, the luciferase reporter gene assay was employed to investigate the functionality of the -98A/C variant identified in the 5’ untranslated region of the LGALS13 gene. Statistically significant differences were observed between patient and control groups at the following loci in the Coloured population: -98A/C, IVS2 -36G/A, IVS2 -22A/G and the hotspot variant in exon 3 (p<0.05). These variants could represent a susceptibility profile of this population or alternatively have implications in the pathogenesis of abruptio placentae. The deletion of a single thymine in exon 3 was shown to result in truncation of PP13 and subsequent disruption of a number of cysteine residues and putative phosphorylation sites, which could impact on dimerization and ultimately, the function of the protein. The reporter gene assay revealed a significant reduction (p=0.004) in luciferase activity by the -98 C allele. iii In silico analysis suggests that this reduction could be due the disruption of a NF1 or GR transcription factor binding site. This study provides evidence that variants in the LGALS13 gene may underlie and/or confer susceptibility to abruptio placentae by impacting on gene regulation or resulting in the expression of an aberrant form of the PP13 which could affect functionality and thereby result in the disruption of normal implantation and placentation.

The genetic architecture of placental abruption (PA) remains poorly understood. We examined variations in SNPs of circadian clock-related genes in placenta with PA risk. We also explored placental and maternal genomic contributions to PA risk. Placental genomic DNA samples were isolated from 280 PA cases and 244 controls. Genotyping was performed using the Illumina Cardio-MetaboChip. We examined 116 SNPs in 13 genes known to moderate circadian rhythms. Logistic regression models were fit to estimate odds ratios (ORs). The combined effect of multiple SNPs on PA risk was estimated using a weighted genetic risk score. We examined independent and joint associations of wGRS derived from placental and maternal genomes with PA. Seven SNPs in five genes (ARNTL2, CRY2, DEC1, PER3 and RORA), in the placental genome, were associated with PA risk. Each copy of the minor allele (G) of a SNP in the RORA gene (rs2899663) was associated with a 30% reduced odds of PA (95% CI 0.52-0.95). The odds of PA increased with increasing placental-wGRS (P_trend<0.001). The ORs were 1.00, 2.16, 3.24 and 4.48 across quartiles. Associations persisted after the maternal-wGRS was included in the model. There was evidence of an additive contribution of placental and maternal genetic contributions to PA risk. Participants with placental- and maternal-wGRS in the highest quartile, compared with those in the lowest quartile, had a 15.57-fold (95% CI 3.34- 72.60) increased odds of PA. Placental variants in circadian clock-related genes are associated with PA risk; and the association persists after control of genetic variants in the maternal genome

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / While there is consistent evidence that episodes of physical exertion are associated with an immediately higher risk of acute ischemic vascular events, the risk of placental abruption immediately following episodes of physical exertion has not been studied. In a multicenter case-crossover study, we interviewed 663 women with placental abruption at 7 Peruvian hospitals between January 2013 and August 2015. We asked women about physical exertion in the hour before symptom onset and compared this with their frequency of physical exertion over the prior week. Compared with times with light or no exertion, the risk of placental abruption was 7.8 (95% confidence interval (CI): 5.5, 11.0) times greater in the hour following moderate or heavy physical exertion. The instantaneous incidence rate ratio of placental abruption within an hour of moderate or heavy physical exertion was lower for women who habitually engaged in moderate or heavy physical activity more than 3 times per week in the year before pregnancy (rate ratio (RR) = 3.0, 95% CI: 1.6, 5.9) compared with more sedentary women (RR = 17.3, 95% CI: 11.3, 26.7; P for homogeneity < 0.001), and the rate ratio was higher among women with preeclampsia/eclampsia (RR = 13.6, 95% CI: 7.0, 26.2) than among women without (RR = 6.7, 95% CI: 4.4, 10.0; P for homogeneity = 0.07). / Revisión por pares / Revisión por pares

Made available in DSpace on 2014-06-11T19:23:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-11-27Bitstream added on 2014-06-13T19:30:08Z : No. of bitstreams: 1 toledo_czp_me_jabo.pdf: 2114280 bytes, checksum: 3353189f0d92e6cea2a99b3bf82c63cc (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Objetivou-se com este estudo analisar a morfologia placentária de fêmeas asininas, espécie, cuja importância no meio rural é de inestimável valor, sendo poucas as informações detalhadas sobre sua placenta, “órgão” imprescindível à viabilidade fetal e a interação materno-fetal, mediante estudo macroscópico, microscópico e de microscopia eletrônica de varredura, os quais, exceto a macroscopia, foram realizados também na interação materno-fetal (útero-placentária). Para tanto, utilizaram-se onze fêmeas desta espécie e foram colhidas 11 placentas logo após a parição a campo, no instante de sua liberação, além de fragmentos de uma placenta oriunda de uma dessas 11 fêmeas, que teve sua gestação acompanhada por duas vezes. Assim, na primeira delas realizou-se operação cesariana e fragmentos da interação materno-fetal foram colhidos; parte deles fixados para posterior processamento usual em histologia e, outra parte, fixada e processada para análise à microscopia eletrônica de varredura. Sua segunda gestação foi acompanhada até o momento da parição e a liberação da placenta após parto normal, sendo colhida na íntegra. Após a colheita, as onze placentas foram pesadas, analisadas macroscopicamente e alguns fragmentos placentários fixados para subseqüente processamento usual em histologia; outros fragmentos foram fixados convenientemente para posterior processamento à microscopia eletrônica de varredura. Das peças analisadas e das preparações microscópicas e ultraestruturais, foram realizadas fotografias, fotomicrografias e ultrafotografias para a documentação. Verificou-se que o valor médio dos pesos dos jumentos, como os pesos das placentas, em relação aos neonatos foi de 8.36.1.10 e 13,18% respectivamente... / The objective of this study was analyzing the placental morphology of female asinines, species whose importance in the rural area is priceless, being little the detailed information about their placenta, vital “organ” to fetal viability and maternal fetal interaction, upon macroscopic, microscopic study, and scanning electron microscopy, which, except the macroscopy, were performed also in the maternal fetal interaction (placental-uterus). Therefore, eleven females of this species were used and eleven placentae were collected right after the calving down at field, in the instant of their release, besides the placenta fragments of one of these eleven females, which had its gestation followed-up twice. Thus, in the first of them, a caesarian operation was performed and fragments of the maternal fetal interaction were collected; part of them fixed for future usual processing in histology and, the other part, fixed and processed for analyses to scanning electron microscopy. Its second gestation was followed up until the moment of calving and the liberation of the placenta after the normal calving, being collected at full. After being collected, the eleven placentae were weighed, analyzed macroscopically and some placental fragments fixed for later usual processing in histology; other fragments were fixed conveniently for future processing to scanning electron microscopy. Among the analyzed pieces and the microscopic and ultra structural preparations, photographs, photomicrographs and ultra photographs were taken for the documentation. It was found that the average weights of the donkeys, as the weights of the placentae, in relation to the neonates were 8.36.1.10 and 13.18% respectively... (Coplete abstract click electronic access below)

Resumo: Objetivou-se com este estudo analisar a morfologia placentária de fêmeas asininas, espécie, cuja importância no meio rural é de inestimável valor, sendo poucas as informações detalhadas sobre sua placenta, "órgão" imprescindível à viabilidade fetal e a interação materno-fetal, mediante estudo macroscópico, microscópico e de microscopia eletrônica de varredura, os quais, exceto a macroscopia, foram realizados também na interação materno-fetal (útero-placentária). Para tanto, utilizaram-se onze fêmeas desta espécie e foram colhidas 11 placentas logo após a parição a campo, no instante de sua liberação, além de fragmentos de uma placenta oriunda de uma dessas 11 fêmeas, que teve sua gestação acompanhada por duas vezes. Assim, na primeira delas realizou-se operação cesariana e fragmentos da interação materno-fetal foram colhidos; parte deles fixados para posterior processamento usual em histologia e, outra parte, fixada e processada para análise à microscopia eletrônica de varredura. Sua segunda gestação foi acompanhada até o momento da parição e a liberação da placenta após parto normal, sendo colhida na íntegra. Após a colheita, as onze placentas foram pesadas, analisadas macroscopicamente e alguns fragmentos placentários fixados para subseqüente processamento usual em histologia; outros fragmentos foram fixados convenientemente para posterior processamento à microscopia eletrônica de varredura. Das peças analisadas e das preparações microscópicas e ultraestruturais, foram realizadas fotografias, fotomicrografias e ultrafotografias para a documentação. Verificou-se que o valor médio dos pesos dos jumentos, como os pesos das placentas, em relação aos neonatos foi de 8.36.1.10 e 13,18% respectivamente... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The objective of this study was analyzing the placental morphology of female asinines, species whose importance in the rural area is priceless, being little the detailed information about their placenta, vital "organ" to fetal viability and maternal fetal interaction, upon macroscopic, microscopic study, and scanning electron microscopy, which, except the macroscopy, were performed also in the maternal fetal interaction (placental-uterus). Therefore, eleven females of this species were used and eleven placentae were collected right after the calving down at field, in the instant of their release, besides the placenta fragments of one of these eleven females, which had its gestation followed-up twice. Thus, in the first of them, a caesarian operation was performed and fragments of the maternal fetal interaction were collected; part of them fixed for future usual processing in histology and, the other part, fixed and processed for analyses to scanning electron microscopy. Its second gestation was followed up until the moment of calving and the liberation of the placenta after the normal calving, being collected at full. After being collected, the eleven placentae were weighed, analyzed macroscopically and some placental fragments fixed for later usual processing in histology; other fragments were fixed conveniently for future processing to scanning electron microscopy. Among the analyzed pieces and the microscopic and ultra structural preparations, photographs, photomicrographs and ultra photographs were taken for the documentation. It was found that the average weights of the donkeys, as the weights of the placentae, in relation to the neonates were 8.36.1.10 and 13.18% respectively... (Coplete abstract click electronic access below) / Orientador: Gilson Hélio Toniollo / Coorientador: Márcia Rita Fernandes Machado / Banca: Marion Burkhardt de Koivisto / Banca: Marcos Lania de Araújo / Mestre

Nos suínos a capacidade uterina influencia o tamanho de leitegada, podendo servir como parâmetro para seleção de animais com melhor desempenho reprodutivo. Evidências destacam que a profundidade da penetração da pipeta correlaciona-se positivamente com o tamanho do útero, servindo de ferramenta para métodos de seleção de leitoas com maior capacidade uterina. Sessenta e seis leitoas pré-púberes de linhagem comercial tiveram a puberdade induzida através de combinação hormonal (eCG e LH). A medida de comprimento da penetração da pipeta de IA foi realizada nas 40 leitoas que apresentaram o 2&ordm cio de maneira sincronizada, sendo que 4 leitoas retornaram o cio. Aos 69 dias de gestação, em média, as 36 leitoas foram abatidas, analisando-se os dados de tamanho de útero, placenta e sobrevivência fetal. O comprimento e peso uterino apresentaram uma correlação positiva significativa com a taxa de sobrevivência gestacional (r=0,43 e r=0,37, respectivamente). Verificou-se que a penetração da pipeta no animal não foi capaz de predizer a capacidade uterina, embora o tamanho uterino tenha influenciado positivamente a taxa de sobrevivência gestacional. A área da placenta correlacionou-se positivamente com o peso da placenta (r=0,66; P<0,0001), e estas com o peso fetal (r=0,60; P<0,0001 e r=0,59; P<0,001, respectivamente). O número de fetos, o comprimento uterino e o peso uterino não se associaram com peso fetal. Fundamentando-se nos resultados obtidos, a penetração da pipeta no momento da inseminação artificial não foi capaz de predizer a capacidade uterina, tampouco sobrevivência dos conceptos aos 70 dias de gestação. / Swines uterine capacity affects litter size, and it could be used as selection parameter of animals with a high reproductive performance. Evidences show that the catheter penetration length is positively correlated with uterine size, and it can be used as selection tool of high uterine capacity in gilts. Sixty six prepubertal gilts of commercial line had the puberty inducted by a hormonal combination (eCG and LH). The Artificial Insemination (AI) catheter penetration length was tested on fourty gilts at the second estrus, but four gilts returned to estrus. The thirty-six remained gilts were slaughtered an average at age sixty-nine of gestation, analyzing the uterus and placental size, and the concepts survival. The uterine length and weight showed a significative and positive correlation with the concepts survival rate (r=0,43 and r=0,37, respectively). The catheter penetration length was not able to predict the uterine capacity, however the uterine size had positively influentied the survival rate. The average of placental area was positively correlated with the average placental weight (r=0,66; P<0,0001), and those ones with the average fetal weight (r=0,60; P<0,0001 and r=0,59; P<0,001, respectively). Analyzing the results obtained, the catheter penetration length measurement during artificial insemination did not predict the uterine capacity, neither the concepts survival on seventy days of gestation age.

Nos suínos a capacidade uterina influencia o tamanho de leitegada, podendo servir como parâmetro para seleção de animais com melhor desempenho reprodutivo. Evidências destacam que a profundidade da penetração da pipeta correlaciona-se positivamente com o tamanho do útero, servindo de ferramenta para métodos de seleção de leitoas com maior capacidade uterina. Sessenta e seis leitoas pré-púberes de linhagem comercial tiveram a puberdade induzida através de combinação hormonal (eCG e LH). A medida de comprimento da penetração da pipeta de IA foi realizada nas 40 leitoas que apresentaram o 2&ordm cio de maneira sincronizada, sendo que 4 leitoas retornaram o cio. Aos 69 dias de gestação, em média, as 36 leitoas foram abatidas, analisando-se os dados de tamanho de útero, placenta e sobrevivência fetal. O comprimento e peso uterino apresentaram uma correlação positiva significativa com a taxa de sobrevivência gestacional (r=0,43 e r=0,37, respectivamente). Verificou-se que a penetração da pipeta no animal não foi capaz de predizer a capacidade uterina, embora o tamanho uterino tenha influenciado positivamente a taxa de sobrevivência gestacional. A área da placenta correlacionou-se positivamente com o peso da placenta (r=0,66; P<0,0001), e estas com o peso fetal (r=0,60; P<0,0001 e r=0,59; P<0,001, respectivamente). O número de fetos, o comprimento uterino e o peso uterino não se associaram com peso fetal. Fundamentando-se nos resultados obtidos, a penetração da pipeta no momento da inseminação artificial não foi capaz de predizer a capacidade uterina, tampouco sobrevivência dos conceptos aos 70 dias de gestação. / Swines uterine capacity affects litter size, and it could be used as selection parameter of animals with a high reproductive performance. Evidences show that the catheter penetration length is positively correlated with uterine size, and it can be used as selection tool of high uterine capacity in gilts. Sixty six prepubertal gilts of commercial line had the puberty inducted by a hormonal combination (eCG and LH). The Artificial Insemination (AI) catheter penetration length was tested on fourty gilts at the second estrus, but four gilts returned to estrus. The thirty-six remained gilts were slaughtered an average at age sixty-nine of gestation, analyzing the uterus and placental size, and the concepts survival. The uterine length and weight showed a significative and positive correlation with the concepts survival rate (r=0,43 and r=0,37, respectively). The catheter penetration length was not able to predict the uterine capacity, however the uterine size had positively influentied the survival rate. The average of placental area was positively correlated with the average placental weight (r=0,66; P<0,0001), and those ones with the average fetal weight (r=0,60; P<0,0001 and r=0,59; P<0,001, respectively). Analyzing the results obtained, the catheter penetration length measurement during artificial insemination did not predict the uterine capacity, neither the concepts survival on seventy days of gestation age.

O desenvolvimento sadio e harmonioso de uma gestação está relacionado diretamente a nutrição fetal equilibrada. Alguns minerais são essenciais por meio das funções desempenhadas no metabolismo, manutenção e crescimento fetal. As carências minerais estão entre os fatores que mais contribuem para a baixa produtividade de rebanho, gerando perdas econômicas significativas. Desse modo estudos que auxiliam na compreensão das razões para a baixa produtividade em rebanhos são importantes. A placenta atua como uma barreira seletiva ao fornecimento adequado de nutrientes durante a gestação. O principal objetivo deste estudo foi determinar a concentração de elementos traço para animais não manipulados (GCO) e clonados (GCL) para períodos gestacionais e obter mapas de distribuição qualitativos para placenta de bovinos GCO e GCL. As amostras de tecido placentário foram selecionadas e agrupadas em três períodos gestacionais: estágio 1, 90 dias; estágio 2, 135 dias e estágio 3, 235 dias. Todas as amostras foram analisadas para os elementos constituintes e traço por Espectrometria de Raios X. Os resultados obtidos nas análises de tecido placentário contendo elementos constituintes e traço indicaram que os procedimentos de preparação foram adequados. Os resultados obtidos indicaram variações significativas para os elementos Na, Mg, P, Cl, K e Ca. Os mapas de distribuição elementar confirmaram que a maior concentração elementar ocorreu na região de interdigitação entre as criptas carunculares do epitélio uterino e as vilosidades cotiledonária com células trofoblásticas na região dos placentônios e nas glândulas endometriais da região interplacentomal, indicando que nestas regiões intensificam-se as trocas de nutrientes entre mãe e feto. Para os resultados obtidos na avaliação dainterelação elementar, quanto as amostras de placentônio bovino clonado observou-se as interelações entre Ca-Fe, Cu-Fe, Zn-Ca, Fe-Zn, Ca-Cu e Cu-Zn. Pelo exposto acima, concluiu-se que as análises químicas foram adequadas para a determinação da concentração de elementos constituintes e traço e que a análise qualitativa dos mapas bidimensionais e mapas de correlação obtidos por meio de experimento com radiação Síncrotron foram essenciais para a discussão sobre as alterações nas regiões selecionadas e períodos de gestação avaliados. / The healthy and harmonious development of a gestation is directly associated to a balanced fetus nutrition. Some minerals are essential because of their functions developed in the metabolism, maintenance and fetus development. Mineral deficiencies are among the factors that contribute the most for a low productivity of the cattle, generating significant economic loss. Thus, studies that help understand the reasons for a low productivity in cattle are important. The placenta works as a selective barrier to the adequate supply of nutrients during the gestation. The aim of this study was to determine the concentration of trace elements for non-manipulative animals (GCO) and cloned animals (GCL) for gestational periods and to obtain maps of qualitative distribution for the placenta of bovines GCO and GCL. Samples of placenta tissues were selected and grouped in three gestational periods: stage 1, 90 days; stage 2, 135 days; and stage 3, 235 days. All the samples were analyzed for the constituent elements and traces by X Rays Spectrometry. The results obtained in the analyses of placenta tissue bearing constituent elements and trace indicated that the procedures of preparation were adequate. The results indicated significant variations for the elements Na, Mg, P, Cl, K e Ca. The maps of elementary distribution confirmed that the highest elementary concentration occurred in the region of inter-digitation of carunculate vaults of the uterine epithelium and cotiledonary vilosities with trophoblast cells, in the regions of the placentons and endometrial glands of the interplacentomal region, confirming that, in these regions, nutrient exchanges intensify between mother and fetus. For the results obtained in evaluation of the elementary interrelation in relation to the samples of cloned bovine placentons we observed that the maps demonstrated several interrelations between Ca-Fe, Cu-Fe, Zn-Ca, Fe-Zn, Ca-Cu, and Cu-Zn. By these results, we concluded that the chemical analyses were adequate for the determination of the elementary concentration of constituent elements and trace and also that the qualitative analysis of the bi-dimensional maps and correlated maps obtained through experiments with Synchrotron radiation were essential for the discussion on alterations on selected regions and on evaluated periods of gestations.

O desenvolvimento sadio e harmonioso de uma gestação está relacionado diretamente a nutrição fetal equilibrada. Alguns minerais são essenciais por meio das funções desempenhadas no metabolismo, manutenção e crescimento fetal. As carências minerais estão entre os fatores que mais contribuem para a baixa produtividade de rebanho, gerando perdas econômicas significativas. Desse modo estudos que auxiliam na compreensão das razões para a baixa produtividade em rebanhos são importantes. A placenta atua como uma barreira seletiva ao fornecimento adequado de nutrientes durante a gestação. O principal objetivo deste estudo foi determinar a concentração de elementos traço para animais não manipulados (GCO) e clonados (GCL) para períodos gestacionais e obter mapas de distribuição qualitativos para placenta de bovinos GCO e GCL. As amostras de tecido placentário foram selecionadas e agrupadas em três períodos gestacionais: estágio 1, 90 dias; estágio 2, 135 dias e estágio 3, 235 dias. Todas as amostras foram analisadas para os elementos constituintes e traço por Espectrometria de Raios X. Os resultados obtidos nas análises de tecido placentário contendo elementos constituintes e traço indicaram que os procedimentos de preparação foram adequados. Os resultados obtidos indicaram variações significativas para os elementos Na, Mg, P, Cl, K e Ca. Os mapas de distribuição elementar confirmaram que a maior concentração elementar ocorreu na região de interdigitação entre as criptas carunculares do epitélio uterino e as vilosidades cotiledonária com células trofoblásticas na região dos placentônios e nas glândulas endometriais da região interplacentomal, indicando que nestas regiões intensificam-se as trocas de nutrientes entre mãe e feto. Para os resultados obtidos na avaliação dainterelação elementar, quanto as amostras de placentônio bovino clonado observou-se as interelações entre Ca-Fe, Cu-Fe, Zn-Ca, Fe-Zn, Ca-Cu e Cu-Zn. Pelo exposto acima, concluiu-se que as análises químicas foram adequadas para a determinação da concentração de elementos constituintes e traço e que a análise qualitativa dos mapas bidimensionais e mapas de correlação obtidos por meio de experimento com radiação Síncrotron foram essenciais para a discussão sobre as alterações nas regiões selecionadas e períodos de gestação avaliados. / The healthy and harmonious development of a gestation is directly associated to a balanced fetus nutrition. Some minerals are essential because of their functions developed in the metabolism, maintenance and fetus development. Mineral deficiencies are among the factors that contribute the most for a low productivity of the cattle, generating significant economic loss. Thus, studies that help understand the reasons for a low productivity in cattle are important. The placenta works as a selective barrier to the adequate supply of nutrients during the gestation. The aim of this study was to determine the concentration of trace elements for non-manipulative animals (GCO) and cloned animals (GCL) for gestational periods and to obtain maps of qualitative distribution for the placenta of bovines GCO and GCL. Samples of placenta tissues were selected and grouped in three gestational periods: stage 1, 90 days; stage 2, 135 days; and stage 3, 235 days. All the samples were analyzed for the constituent elements and traces by X Rays Spectrometry. The results obtained in the analyses of placenta tissue bearing constituent elements and trace indicated that the procedures of preparation were adequate. The results indicated significant variations for the elements Na, Mg, P, Cl, K e Ca. The maps of elementary distribution confirmed that the highest elementary concentration occurred in the region of inter-digitation of carunculate vaults of the uterine epithelium and cotiledonary vilosities with trophoblast cells, in the regions of the placentons and endometrial glands of the interplacentomal region, confirming that, in these regions, nutrient exchanges intensify between mother and fetus. For the results obtained in evaluation of the elementary interrelation in relation to the samples of cloned bovine placentons we observed that the maps demonstrated several interrelations between Ca-Fe, Cu-Fe, Zn-Ca, Fe-Zn, Ca-Cu, and Cu-Zn. By these results, we concluded that the chemical analyses were adequate for the determination of the elementary concentration of constituent elements and trace and also that the qualitative analysis of the bi-dimensional maps and correlated maps obtained through experiments with Synchrotron radiation were essential for the discussion on alterations on selected regions and on evaluated periods of gestations.