Made available in DSpace on 2014-08-13T14:50:44Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-25Bitstream added on 2014-08-13T18:00:48Z : No. of bitstreams: 1 000753371.pdf: 1423081 bytes, checksum: e2ae663efebbbdf0a3923a02cb14bd38 (MD5) / A pré-eclâmpsia (PE) é uma síndrome específica da gestação humana, caracterizada pelo aparecimento de hipertensão arterial e proteinúria após a 20ª. semana de gestação. É aceito que esta patologia tem origem na placenta, provavelmente em decorrência de fatores envolvidos na sua formação e desenvolvimento. O presente projeto teve como objetivo: 1) Avaliar comparativamente a histopatologia de lesões placentárias em gestantes portadoras de PE e gestantes normotensas; 2) Comparar a frequência de recém-nascidos com restrição de crescimento intrauterino entre os grupos de gestantes estudados. Foram avaliadas 140 gestantes, sendo 20 normotensas e 120 com PE. As gestantes com PE foram classificadas de acordo com o aparecimento das manifestações clínicas em PE precoce (< 34 semanas de gestação - n=40) e PE tardia (≥ 34 semanas de gestação – n= 80). Um fragmento de placenta foi obtido imediatamente após o parto e preparado para análise histopatológica, sendo os cortes histológicos de 4m de espessura colocados sobre lâmina histológica para coloração pelo método de Hematoxilina - Eosina (HE). O exame histológico das placentas mostrou que as patologias mais frequentemente encontradas em gestantes com PE foram aumento de vilos com nós sinciciais, infarto, aumento de depósito de fibrina e hipoplasia de vilos distais. Placentas de gestantes portadoras de PE precoce apresentaram maior percentagem de vilos com aumento de nós sinciciais para a idade gestacional em comparação a gestantes normotensas e com PE tardia. O infarto placentário e a hipoplasia de vilos distais foram mais frequentes em gestantes portadoras de PE quando comparadas às normotensas, enquanto o depósito de fibrina foi maior em gestantes com PE precoce do que nas com PE tardia. Além disso, a percentagem de recém-nascidos com restrição de crescimento intrauterino foi significativamente maior (37,8%) em gestantes com PE precose ... / Preeclampsia (PE) is a specific syndrome of human pregnancy, characterized by the onset of hypertension and proteinuria after the 20th week of gestation. It is accepted that this disease originates in the placenta, probably due to factors involved in its formation and development. The objectives of the present study were: 1) To evaluate the histopathology of placental lesions in pregnant women with PE and in normotensive pregnant women; 2) To compare the frequency of newborns with intrauterine growth restriction among the groups of pregnant women studied. One hundred and forty pregnant women were evaluated, of whom 20 were normotensive and 120 were preeclamptic. Pregnant women with PE were classified according to the appearance of clinical manifestations in early-onset PE (<34 weeks of gestation - n = 40) and late-onset PE (≥ 34 weeks gestation - n = 80). A fragment of placenta was obtained immediately after delivery and prepared for histopathological analysis. Placental sections of 4m were placed on a slide for hematoxylin - eosin (HE) staining. Histological examination of placentas showed that the most frequently alterations found in patients with PE were increase villous with syncytial knots, infarction, increased fibrin deposits and distal villi hypoplasia. Placenta of pregnant women with early-onset PE showed higher percentage of villous with syncytial knots for gestation age compared with late-onset PE and normotensive pregnant women. Infarct and distal villi hypoplasia were more frequent in placenta of pregnant women with PE, whereas fibrin deposits were significantly increased in women with early-onset PE than in late-onset PE. Moreover, the percentage of newborns with intrauterine growth restriction was higher (37.8%) in patients with early–onset PE than in late-onset PE (14.3%). Together the results show that pregnant women with early-onset PE showed placentas with higher percentage of lesions and newborns with ...

Made available in DSpace on 2015-01-26T13:21:21Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-08-08Bitstream added on 2015-01-26T13:30:38Z : No. of bitstreams: 1 000797817.pdf: 449554 bytes, checksum: 7adc756b2ac04e5d1cbc6be34fa7a04e (MD5) / Introdução: A síndrome HELLP é uma complicação severa da pré-eclâmpsia, caracterizada por hemólise, elevação das enzimas hepáticas e trombocitopenia. Apesar de haver padronização dos valores laboratoriais que definem a síndrome HELLP, ainda existe dificuldade para a caracterização da hemólise. OBJETIVO: Avaliar o valor de haptoglobina que determina a hemólise nas pacientes com síndrome HELLP. MÉTODOS: estudo transversal e prospectivo de gestantes e puérperas com de pré-eclâmpsia. Exame laboratorial avaliado: dosagem sérica de haptoglobina. Construção da curva ROC para determinar o valor de corte da haptoglobina para diagnóstico de hemólise na síndrome HELLP. RESULTADOS: O valor da haptoglobina para diagnóstico de hemólise em pacientes com síndrome HELLP foi de 0,26g/L. DISCUSSÃO: A melhor correlação observada foi a haptoglobina com a DHL, indicando que este é o melhor marcador de hemólise intravascular para o diagnóstico da síndrome HELLP. CONCLUSÃO: A dosagem sérica da haptoglobina nos casos de pré-eclâmpsia deve fazer parte dos exames de rotina para diagnóstico de hemólise intravascular da síndrome HELLP / Context: HELLP syndrome is a severe complication of pre-eclampsia, caracterized by hemolysis, elevated liver enzymes and low platelet count. Although there are standardized laboratory values that define the HELLP syndrome, the difficulty still exists for the characterization of hemolysis. PURPOSE: to evaluate the haptoglobin value to diagnose the hemolysis in HELLP syndrome. METHODS: transversal and prospective study of pregnant and postdelivery women with pre-eclampsia. Laboratory tests evaluated: serum haptoglobin. ROC curve to determine the cutoff value of haptoglobin in the diagnosis of hemolysis in HELLP syndrome. RESULTS: The haptoglobin value for hemolysis diagnosis in HELLP syndrome was 0.26 g / L. DISCUSSION: The best correlation was with haptoglobin and DHL, indicating that this is the best marker of intravascular hemolysis for the diagnosis of HELLP syndrome. CONCLUSION: Serum haptoglobin in cases of pre-eclampsia should be part of routine tests for diagnosis of intravascular hemolysis in HELLP syndrome

Made available in DSpace on 2014-06-11T19:29:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-08-26Bitstream added on 2014-06-13T18:39:55Z : No. of bitstreams: 1 amaral_wt_me_botfm.pdf: 465005 bytes, checksum: 4d438b3030a97d550ee1de745b48a539 (MD5) / Fundação de Ensino e Pesquisa em Ciências da Saúde (FEPECS) / A pré-eclâmpsia é uma das principais causas de morbimortalidade materna e perinatal no Brasil e no mundo. É uma patologia heterogênea, multifatorial, sem etiologia esclarecida e fisiopatologia complexa. A identificação de fatores de risco ao seu desenvolvimento pode auxiliar na prevenção e diagnóstico precoce do início clínico da doença. Identificar fatores de risco e complicações da pré-eclâmpsia (PE) / hipertensão gestacional (HG) em uma população de primigestas atendida no Hospital Regional da Ceilândia – Brasília - Distrito Federal. Foi realizado estudo caso controle, no período de novembro de 2009 a dezembro de 2010, em população de primigestas com resolução da gestação no Hospital Regional da Ceilândia (DF). Foram incluídas todas as primigestas com diagnóstico de PE / HG, com escolha aleatória dos casos controle (01 primigesta com PE/HG para 03 primigestas normotensas). As variáveis estudadas (sócio-demográficas, obstétricas, complicações maternas e perinatais) foram coletadas através de questionário previamente testado, após o consentimento livre e esclarecido da gestante. Na análise dos dados foram aplicados os testes do qui-quadrado, de Fisher e de comparação múltipla. Dentre 1264 primigestas analisadas, 972 (76,9%) eram normotensas e 292 (23,1%) hipertensas, destas 64 (22%) eram hipertensas gestacionais e 228 (78%) pré-eclâmpticas. Fatores de risco tais como idade materna acima de 35 anos, mãe hipertensa ou com hipertensão durante a gestação, irmã com hipertensão na gestação, uso de condon masculino como meio anticoncepcional, obesidade e ganho de peso excessivo durante a gestação mostraram correlação estatística com o desenvolvimento da patologia. O desenvolvimento da doença em primíparas está associado à história familiar e à obesidade. Fatores que podem servir como screening para identificação precoce de pacientes em risco de desenvolverem pré-eclâmpsia / Preeclampsia is a major cause of morbidity and mortality maternal and perinatal in Brazil and worldwide. It is a heterogeneous, multifactorial disorder. The aim of this study was to determine the risk factors associated with preeclampsia. We hope that these factors can be used as a screening for preeclampsia prediction. To analyze population of primiparous pregnant women diagnosed with Preeclampsia / Gestational Hypertension, in Ceilândia Hospital (HRC) – Brasília –Distrito Federal. Case-Control Study from November 2009 to December 2010 in a population of primiparous mothers who give birth at Ceilândia Hospital (DF). Will be included all women diagnosed with pregnancy and Preeclampsia / Gestational Hypertension (National High Blood Pressure Education Program, 2000), with random selection of control cases (01 with postpartum HA - 03 normotensive mothers. Minimum Forecast: 1200). The patient must have a diagnosis of pregnancy, first pregnancy. Data will be collected through a previously tested questionnaire; consent was obtained from the patient. Among 1264 mothers, primiparous women who gave birth in the HRC between november/2009 December/2010, 972 were diagnosed as normotensive and 292 hypertensive, 64 PE and 228 non-proteinuric gestational hypertension. Epidemiological data such as maternal age above 35 years, mother hypertension or with hypertension during pregnancy, sister with hypertension in pregnancy, use of male condom as a mean of contraception, obesity and excessive weight gain during pregnancy showed a strong statistical correlation with the development PE. The development of disease in nulliparous women is associated with family history, obesity. Factors that could serve as screening for early identification of patients at risk for developing preeclampsia

Estudo realizado na Clínica Obstétrica da Faculdade de Medicina da Universidade de São Paulo, com seiscentos e quarenta e cinco gestantes recrutadas, prospectivamente, para avaliações dopplervelocimétricas das artérias uterinas, por via endovaginal, entre 11 e 13+6 semanas e entre 20 e 24+6 semanas. A partir de um grupo de 344 casos com desfecho normal da gestação, valores de referências para os índices dopplervelocimétricos médios foram estabelecidos, e estes foram significativamente maiores na primeira avaliação do que na segunda. Além disso, os valores se correlacionaram de forma positiva e significativa (IP r= 0,42, IR r= 0,42, AB r= 0,29, p<0,0001). Incisura uterina bilateral foi encontrada em 43,9% dos casos no primeiro exame e 4,4% na segunda etapa. Também foram descritos os valores de sensibilidade, especificidade, valores preditivos, razão de verossimilhança e risco relativo de diferentes parâmetros dopplervelocimétricos para predição da pré-eclâmpsia, diagnosticada em 25 casos. Entre 11 e 13 sem. + 6 dias, as áreas sob as curvas de caracterísiticas operacionais dos três índices dopplervelocimétricos foram de 0,51. A maioria dos achados dopplervelocimétricos, nesta fase da gestação, não identificou gestações com risco significativamente aumentado para pré-eclâmpsia. Já, entre 20 e 24 sem. + 6 dias, as áreas sob as curvas de características operacionais foram maiores (IP= 0,66, IR= 0,65, AB= 0,65) e o grupo com índices dopplervelocimétricos acima do percentil 85 e/ou incisura bilateral apresentou risco significativamente aumentado, para o posterior surgimento de pré-eclâmpsia na gestação. Entretanto, a sensibilidade e o valor preditivo positivo foram baixos, e não encorajam o uso desse método para predição da doença hipertensiva específica da gestação em nossa população. / At São Paulo University Medical School, six hundred and forty five pregnant women were prospectively recruited for a longitudinal study involving transvaginal uterine artery Doppler assessment at 11?13+6 weeks and 20?24+6 weeks. Reference values for mean uterine artery Doppler indices were established from 344 cases with normal pregnancy outcome. Values found in the first examination were significantly higher and positively correlated to values obtained in the second examination (PI r= 0.42, RI r= 0.42, SD r= 0.29, p<0.0001). Bilateral notches were found in 43.9% of the cases examined between 11 and 13 weeks, and 4.4% of the cases in the second assessment. Twenty-five cases subsequently developed pre-eclampsia. Sensitivity, specificity, positive and negative predictive values, likelihood ratios and relative risks were calculated for various uterine artery Doppler findings. Between 11 and 13+6 weeks, the ROC curve area was 0.51 for all three indices. At this stage, most uterine artery Doppler findings were not associated with increased risk of pre-eclampsia. At 20 to 20+6 weeks, ROC curve areas were higher (PI= 0.66, RI= 0.65, SD= 0.65) and increased impedance to flow (above the 85th centile) and/or bilateral notches were associated with a significant increase of the risk for the subsequent development of pre-eclampsia. However low sensitivity and positive predictive values do not support this as a screening method for pre-eclampsia in our population.

Introdução: A CALPAIN5faz parte da família das cisteínas proteases e está relacionada com a regulação de inúmeras funções celulares, entre elas a diferenciação e a apoptose. Estudos com microarrays identificaram a CALPAIN5como um alvo da açãotranscripcional do HOXA10em úteros de camundongos. Objetivos: No presente estudo avaliou-se a regulação da CALPAIN5pelo HOXA10 em células endometriais, a expressão da CALPAIN5no endométrio durante todo o ciclo menstrual, e na decídua do primeiro e terceiro trimestres de gestações normais, e o padrão de expressão anormal desse gene na pré-eclampsia e endometriose. Material e Métodos: Foram obtidas dez biópsias endometriais (cinco na fase proliferativa e cinco na fase secretora) depacientes férteis. Biópsias de lesões de endometriose, confirmadas por histopatologia,foram retiradas de dez mulheres durante procedimento vídeo-laparoscópico. Amostras de decídua foram coletadas em três diferentes ocasiões: três amostras do primeiro trimestre,cinco amostras no terceiro trimestre e cinco amostras de pacientes com pré-clâmpsia no terceiro trimestre. Identificou-se a proteína da CALPAIN5utilizando imunohistoquimica (IHC) no endométrio eutópico, ectópico e na decídua. Os resultados da IHC foram quantificados, a partir dedois diferentes observadores, utilizando-se Hscorepara células estromais, epiteliais e deciduais. Transfeccionou-se 4,0µg de pcDNA/HOXA10e 20µM siRNA/HOXA10em cultura de células estromais endometriais humanas (HESC) e células epiteliais endometriais humanas (Ishikawa), assim como os seus respectivos controles. A transfecção foi realizada em quadruplicata quando a cultura de células apresentava confluência de 60-70%. Após 48 horas do procedimento o RNA foi extraído, que deu origem a DNA complementar e após uma reação de cadeia da polimerase em tempo real (PCR em tempo real) foi realizada, em triplicata, para determinar o padrão de expressão do HOXA10e CALPAIN5. A análise estatística foi realizada utilizando-se os testes ANOVA com test pos-hocpara o Hscoree teste tpara os resultados da PCR em tempo real. Resultados: CALPAIN5 é expressadurante todo o ciclo menstrual tanto nas células estromais como epiteliais glandulares, e está mais expressa na decídua do primeiro trimestre. Na endometriose CALPAIN5 se mostrou pouco evidente em ambas as células endometriais (estromais e glandulares), quando comparadas de forma geral com as células endometriais eutópicas das pacientes controles, sendo que sua expressão reduziu em 50% (p<0,05). Nas decíduas de pacientes com pré-eclampsia CALPAIN5 apresentou-se mais expressa que no grupo controle (p<0,05). Nas célulasestromais endometriais o pcDNA/HOXA10aumentou a expressão da CALPAIN5em 11 vezes (p<0,05) e a transfecção com siRNA/HOXA10reduziu a expressão da CALPAIN5em 23 vezes (p<0,05). Conclusão: O HOXA10regula a expressão genética da CALPAIN5nas células endometriais. CALPAIN5 é expressa no endométrio normal e tem sua expressão aumentada nas células deciduais do primeiro trimestre em relação àsfases do ciclo menstrual. CALPAIN5 está pouco expressa no endométrio ectópico na endometriose e mais expressa nas células deciduais na pré-eclampsia grave quando comparados com seus respectivos controles. / BACKGROUND: CALPAIN5is member of the calpain-like cystein protease family and has been implicated in the regulation of a variety of cellular functions, including differentiation and apoptosis. Research with microarray screen identified CALPAIN5as target of HOXA10 transcriptional control in murine endometrium. OBJECTIVES: We propose in this study to demonstrate regulated CALPAIN5expression in endometrium, and 3rd trimester deciduas and an abnormal expression pattern of this gene in pre eclampsia and endometriosis . MATERIALS & METHODS: Ten endometrial biopsies (5 proliferative phase and 5 secretory phase) were obtained from fertile controls. Histologically confirmed biopsies of endometriosis were obtained from 10 women atthe time of laparoscopy. Five third trimester decidual samples and 3 first trimester deciduas samples from controls, and five 3rd trimester decidual samples from women with pre-eclampsia were obtained at the time of labor. Immunohistochemistry (IHC) was used to identify CALPAIN5protein in eutopic and ectopic endometrium as well as in decidua. IHC was performed with CALPAIN5polyclonal antibody. IHC results were assessed by 2 evaluators blinded to the study groups, and H-SCORES were determined for the stroma, glands and decidua. The human endometrial stromal cell line, HESC, the human endometrial epithelial cell line, Ishikawa were transfected with either 4.0µg pcDNA/HOXA10or 20µM HOXA10siRNA; transfection with empty pcDNA vector or nonspecific siRNA served as respective controls. Cells were transfected in quadruplicate at 60-70% confluence. Forty eight hours after the transfection total RNA was isolated. qRT-PCR was performed in duplicated to determine expression levels of HOXA10and CALPAIN5in each group. Statistical analysis was performed with ANOVA test pos-hocto Hscoreand ttest for real time PCRRESULTS: CALPAIN5was expressed in endometrial stromal and glandular cells throughout the menstrual cycle, and increased expression was noted in the first trimester decidua phase. There was a decrease in CALPAIN5expression in both stromal and glandular cells in endometriosis to 50% ofthat seen in fertile controls (p<0.05). CALPAIN 5 was also expressed in 3rd trimester decidua obtained from pre-eclamptics at higher levels than 3rd trimester control decidua (p<0.05). The regulatory relationship between HOXA10 and CALPAIN5was established by transient transfection analysis. CALPAIN5 gene expression increased 11-fold (p<0.05) after pcDNA/HOXA10transfection of the HESC, and decreased 23-fold (p<0.05) after HOXA10siRNA treatment. CONCLUSIONS: CALPAIN5 expression is regulated by HOXA10. CALPAIN5is expressed in normal endometrium and at upregulated in the decidua ofwomen with pre-eclampsia. CALPAIN5 expression is decreased in endometriosis compared to eutopic endometrium.

Abstract The incidence of pre-eclampsia (PE) is 2–7% in different populations and in the worst cases PE may threaten the survival of both mother and newborn; its pathogenesis is not resolved. Field literature today considers PE an angiogenic disorder. Coordinated vascularization is essential for placental development. We wanted to find novel factors in the etiology of PE, and focused our attention on angiogenesis, inherited thrombophilia and folate-homocysteine metabolism. Homocysteine inhibits endothelial cell proliferation, which is closely related to angiogenesis. We performed gene expression profiling of placental tissue using microarray chips, studied the prevalence of factor V Leiden (FVL), prothrombin (F5) G20210A and methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism in patients with severe pregnancy complications and normal controls, compared the expression of the placental adiponectin, leptin and their receptor genes and the relationship of each to trophoblast apoptosis and further, studied the effect of folic acid fortified mineral water on plasma homocysteine concentration during pregnancy. Gene expression profiling revealed downregulation of nine and upregulation of four genes. Interestingly, in one PE patient with cord compression during delivery the profile resembled that observed in normals. The expression level of the leptin and the adiponectin receptor 1 (ADIPOR1) genes was significantly higher in PE. No other significant expression changes were observed. The rate of apoptosis was higher in patients with PE. The FVL prevalence was 9.5%, in PE cases and 1.8% in the controls; a difference of 7.7%, (95% CI 2.0–13.4%). No statistical difference was found in other polymorphisms.. Maternal serum folate concentration increased in our intervention group, but decreased in the control group (p &lt; 0.05). The plasma homocysteine concentrations decreased more in the intervention group (p &lt; 0.001). The expression of angiogenesis-related placental genes can be altered in PE and cord compression cases. The activity of adipocytokine genes in PE may mean that they have a role in placental angiogenesis and apoptosis. Women with FVL may have an increased risk of PE. Fortified mineral water will help us to ensure that especially pregnant women achieve adequate folate intake.

Pregnancy in women with hemoglobinopathy has been associated with an increased risk of adverse pregnancy outcomes. We conducted a historical cohort study using Discharge Abstract Database for the fiscal year 1991-1992 through 2007-2008. We estimated the frequency of pregnant women with hemoglobinopathy and examined their associations with adverse pregnancy outcomes. Women with sickle cell disease are more likely to develop pre-eclampsia and preterm labor, and to undergo cesarean delivery than women with nutritional deficiency anemia, suggesting that there are other mechanisms beyond anemia that may be responsible for an increased risk of adverse pregnancy outcomes. The data suggested a synergistic effect of hemoglobinopathy and pre-eclampsia on preterm labor and cesarean delivery. Prediction models for pre-eclampsia, preterm labor and cesarean delivery were created and internally validated for women with hemoglobinopathy, with satisfactory discrimination and calibration.

Background: Pre-eclampsia (PET) is one of the major causes of maternal and neonatal morbidity and mortality1. Misclassification of PET can lead to biased or erroneous results in epidemiologic studies resulting in false conclusions. Objectives: The objectives of this thesis are to determine the validity of PET diagnosis in pregnant women in administrative database using the ICD-10-CA codes, to explore the nature of misclassification, and to estimate whether misclassification of PET diagnosis in administrative database may result in biased conclusions. Methods: Pregnant women who participated in the Ottawa and Kingston (OaK) Birth Cohort study and delivered in the Ottawa Hospital were included in the study. All cases with hypertensive disorder of pregnancy in the study population were adjudicated to confirm diagnosis of PET. This adjudicated dataset was used as a reference standard. The PET incidence in hospital discharge database was compared with PET incidence calculated from the reference standard database. Results: 2887 of the requested charts were available for review. The PET incidence was much lower in administrative database (1.47%) than in the OaK Birth Cohort Study (3.6%). The results of the study demonstrated that hospital discharge database via ICD-10-CA was not very sensitive to determine incidence of PET since sensitivity of ICD-10-CA diagnostic codes for PET was low (35.92% with 95% Confidence Intervals (CI): 26.7; 45.9) but specificity, PPV, and NPV were high. The majority of misclassified cases belonged to the category (according to the proposed classification) “PET pregnancies coded with incorrect ICD-10-CA code” (78.88%) followed by the category “Pregnancies affected by PET coded as normal” (14.08%). Conclusion: Using hospital discharge database and ICD-10-CA coding to determine incidence of PET in certain settings may yield low sensitivity. Researchers should validate the results when using the hospital discharge database for PET research to ensure that the findings based on analyses of such data demonstrate what they claimed to demonstrate.

Pre-eclampsia (PE) is a serious disease affecting approximately 5% of pregnancies per annum. The disease etiology is complex but its origin lies in abnormal placental development and function. PE is associated with inflammation, increased nitrative stress and abnormal renewal of syncytiotrophoblast (STB), the transporting epithelium of the placenta. STB is renewed by cytotrophoblasts (CTBs) that proliferate, differentiate and fuse with STB and this is balanced by apoptosis. The amino acid taurine facilitates proliferation, differentiation and apoptosis in non-placental tissues. Taurine is also cytoprotective, protecting cells from damage by inflammatory cytokines. Taurine is transported from maternal blood into STB by the amino acid transporter TauT. In isolated STB membranes, TauT activity is inhibited by agents that nitrate tyrosine residues. This thesis tested the hypothesis that STB TauT activity is down-regulated in PE due to post-translational modification of TauT through tyrosine nitration which lowers intracellular taurine and contributes to altered STB renewal. Placentas were collected from normal pregnancy (NP) and PE (blood pressure >140/90mmHg after 20 weeks gestation in previously normotensive women plus proteinuria >300 mg/L in a 24-hour collection). STB TauT activity, measured as Na+-dependent uptake of 3H-taurine into placental villous fragments, was significantly lower in PE (n=24) compared to NP (n=44). Western blotting of membrane enriched homogenates showed that TauT protein expression (normalised to β-actin) was significantly higher in placentas from PE (n=8) compared to NP (n=9). The presence of nitrotyrosine residues (marker of nitrative stress) in placentas of women with PE and NP was assessed by immunohistochemistry (IHC). The intensity of STB nitrotyrosine staining was greater in PE placentas that had reduced TauT activity (n=8) than in NP (n=7). To determine the effect of nitrative stress on TauT activity and STB renewal, placental villous explants from NP were cultured (7 days; n=6) and treated with SIN-1 (1mM; days 5,6) to induce nitrative stress. STB nitrotyrosine (IHC) and TauT activity (3H-taurine uptake) was determined on day 7 and STB renewal was assessed by IHC for apoptosis (M30), proliferation (dual staining for Ki67 and the CTB marker E-cadherin) and STB integrity (cytokeratin 7). SIN-1 increased STB nitrotyrosine staining intensity compared to controls, confirming induction of nitrative stress. SIN-1 reduced STB TauT activity, increased apoptosis, reduced CTB proliferation and altered STB regeneration compared to control. To determine the effect of reducing intracellular taurine on STB renewal, villous explants were cultured for 7 days with 2.5mM β-alanine to competitively inhibit taurine uptake (n=6). At day 7, intracellular taurine, measured as the steady-state accumulation of 3H-taurine, was 15% of normal. STB turnover was assessed at day 7 as described above. β-alanine significantly increased apoptosis and altered STB regeneration compared to controls. Following statistical analysis all p <0.05.In conclusion, STB TauT activity was lower, and protein expression higher, in PE compared to NP. STB nitrotyrosine was elevated in PE and nitrative stress inhibited STB TauT activity and disrupted STB renewal in vitro. Reducing intracellular taurine also disrupted STB renewal in vitro. Overall the data support the hypothesis that post-translational modification of TauT by nitration inhibits TauT activity in PE. This reduces intracellular taurine which contributes to abnormal renewal of STB. Further work is needed (a) to confirm that TauT is nitrated in PE and that reduced STB TauT activity lowers intracellular taurine and reduces taurine delivery to the fetus and (b) to determine the mechanism/s by which taurine regulates CTB apoptosis and facilitates renewal of STB.

Objetivo: Determinar la asociación entre el hiperinsulinismo (HI) y embarazo en mujeres con síndrome de ovario poliquístico (SOP). Diseño: Revisión Sistemática y Meta-Análisis de estudios observacionales que evaluaron la asociación entre hiperinsulinismo y desenlaces obstétricos. Se realizó la búsqueda de la literatura a través de las bases PubMed-MEDLINE, Web of Science, Embase, Scopus y Cochrane hasta junio del 2015. Paciente(s): Mujeres en edad reproductiva con síndrome de ovario poliquístico. Principales medidas del resultado: El desenlace primario fue embarazo; los desenlaces secundarios fueron aborto espontaneo, diabetes gestacional y preeclampsia. Para el análisis del estudio se utilizó el modelo de efectos aleatorios. Resultados: Encontramos siete cohortes prospectivas y cuatro estudios de casos y control (n=711). La proporción de embarazo en las mujeres con hiperinsulinemia fue significativamente menor que en las mujeres sin hiperinsulinemia (45/213 [21.1%] vs 114/273 [41.8%], OR 0.36, IC 95% 0.21-0.62, P<0.001). No hubo diferencia en porcentaje de abortos entre aquellas con hiperinsulinemia y aquellas sin hiperinsulinemia (9/100 [9.0%] vs 15/147 [10.2%], OR 0.90, IC 95% 0.36-2.22, P˃0.001). La información fue escasa respecto a los efectos de en diabetes gestacional y pre-eclampsia. Conclusiones: Existe asociación significativa e inversa entre hiperinsulinismo y embarazo en mujeres con síndrome de ovario poliquístico. Asimismo, no se encontró diferencias entre pacientes SOP con HI y pacientes SOP sin HI respecto al riesgo de aborto. Por otro lado, no hubo información suficiente para analizar los desenlaces diabetes gestacional y preeclampsia. / OBJECTIVE: To determine the association between hyperinsulinism (HI) and pregnancy in women with polycystic ovarian syndrome (PCOS). PATTERN: Systematic Review and Meta-Analysis of observational studies that evaluated the association between hyperinsulinism and obstetric outcomes. We searched the literature through the databases of PubMed-MEDLINE, Web of Science, Embase, Scopus and Cochrane databases until June 2015. PATIENT(s): Women on reproductive age with polycystic ovarian syndrome. MAIN EFFECTS OF RESULTS: The primary outcome was pregnancy; The secondary outcomes were miscarriage, gestational diabetes and preeclampsia. For the analysis of the study we used the random effects model. RESULTS: We found 7 prospective cohorts and 4 case-control studies (n = 711). The percentage of pregnancy in women with hyperinsulinemia was significantly lower than in women without hyperinsulinemia (45/213 [21.1%] vs 114/273 [41.8%], or 0.36, 95% ci 0.21-0.62, p <0.001). There was no difference in the percentage of miscarriages between women with hyperinsulinemia and women without hyperinsulinemia (9/100 [9.0%] vs 15/147 [10.2%], or 0.90, 95% ci 0.36-2.22, p0.001). The information was scarce regarding the effects of gestational diabetes and pre-eclampsia. Conclusions: There is a significant and inverse association between hyperinsulinism and pregnancy in women with PCOS. Likewise, we do not found differences between PCOS patients with HI and PCOS patients without HI regarding the risk of miscarriage.On the other hand, there was not enough information to analyze the results of gestational diabetes and preeclampsia. / Tesis