Fetal Alcohol Spectrum Disorder Primary Prevention through FASD Diagnosis: Identification of High-Risk Birth Mothers through the Diagnosis of their Children. Follow-up Study

Thesis (Master's)--University of Washington, 2016-08

Human Fetal Progenitor Tenocytes for Regenerative Medicine.

Tendon injuries are very frequent and affect a wide and heterogeneous population. Unfortunately, the healing process is long with outcomes that are not often satisfactory due to fibrotic tissue appearance, which leads to scar and adhesion development. Tissue engineering and cell therapies emerge as interesting alternatives to classical treatments. In this study, we evaluated human fetal progenitor tenocytes (hFPTs) as a potential cell source for treatment of tendon afflictions, as fetal cells are known to promote healing in a scarless regenerative process. hFPTs presented a rapid and stable growth up to passage 9, allowing to create a large cell bank for off-the-shelf availability. hFPTs showed a strong tenogenic phenotype with an excellent stability, even when placed in conditions normally inducing cells to differentiate. The karyotype also indicated a good stability up to passage 12, which is far beyond that necessary for clinical application (passage 6). When placed in coculture, hFPTs had the capacity to stimulate human adult tenocytes (hATs), which are responsible for the deposition of a new extracellular matrix during tendon healing. Finally, it was possible to distribute cells in porous or gel scaffolds with an excellent survival, thus permitting a large variety of applications (from simple injections to grafts acting as filling material). All of these results are encouraging in the development of an off-the-shelf cell source capable of stimulating tendon regeneration for the treatment of tendon injuries.

Imaging of activated complement using ultrasmall superparamagnetic iron oxide particles (USPIO) - conjugated vectors: an in vivo in utero non-invasive method to predict placental insufficiency and abnormal fetal brain development.

In the current study, we have developed a magnetic resonance imaging-based method for non-invasive detection of complement activation in placenta and foetal brain in vivo in utero. Using this method, we found that anti-complement C3-targeted ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles bind within the inflamed placenta and foetal brain cortical tissue, causing a shortening of the T2* relaxation time. We used two mouse models of pregnancy complications: a mouse model of obstetrics antiphospholipid syndrome (APS) and a mouse model of preterm birth (PTB). We found that detection of C3 deposition in the placenta in the APS model was associated with placental insufficiency characterised by increased oxidative stress, decreased vascular endothelial growth factor and placental growth factor levels and intrauterine growth restriction. We also found that foetal brain C3 deposition was associated with cortical axonal cytoarchitecture disruption and increased neurodegeneration in the mouse model of APS and in the PTB model. In the APS model, foetuses that showed increased C3 in their brains additionally expressed anxiety-related behaviour after birth. Importantly, USPIO did not affect pregnancy outcomes and liver function in the mother and the offspring, suggesting that this method may be useful for detecting complement activation in vivo in utero and predicting placental insufficiency and abnormal foetal neurodevelopment that leads to neuropsychiatric disorders.

Fetal-maternal interface impedance parallels local NADPH oxidase related superoxide production

Blood flow assessment employing Doppler techniques is a useful procedure in pregnancy evaluation, as it may predict pregnancy disorders coursing with increased uterine vascular impedance, as pre-eclampsia. While the local causes are unknown, emphasis has been put on reactive oxygen species (ROS) excessive production. As NADPH oxidase (NOX) is a ROS generator, it is hypothesized that combining Doppler assessment with NOX activity might provide useful knowledge on placental bed disorders underlying mechanisms. A prospective longitudinal study was performed in 19 normal course, singleton pregnancies. Fetal aortic isthmus (AoI) and maternal uterine arteries (UtA) pulsatility index (PI) were recorded at two time points: 20-22 and 40-41 weeks, just before elective Cesarean section. In addition, placenta and placental bed biopsies were performed immediately after fetal extraction. NOX activity was evaluated using a dihydroethidium-based fluorescence method and associations to PI values were studied with Spearman correlations. A clustering of pregnancies coursing with higher and lower PI values was shown, which correlated strongly with placental bed NOX activity, but less consistently with placental tissue. The study provides evidence favoring that placental bed NOX activity parallels UtA PI enhancement and suggests that an excess in oxidation underlies the development of pregnancy disorders coursing with enhanced UtA impedance.

A decrease in dkk1, a Wnt inhibitor, contributes to placental and fetal lipid accumulation in an obesity-prone rat model

Placenta, as the sole transport mechanism between mother and fetus, links the maternal physical state and the immediate and life-long outcomes of the offspring. The present study examined the mechanisms behind the effect of maternal obesity on placental lipid accumulation and metabolism. Pregnant Obese Prone (OP) and Obese Resistant (OR) rat strains were fed a control diet throughout gestation. Placentas were collected on gestational d21 for analysis and frozen placental sections were analyzed for fat accumulation as well as β-Catenin and Dkk1 localization. Additionally, DKK1 was overexpressed in JEG3 trophoblast cells, followed by treatment with NEFA and Oil Red O stain quantification and mRNA analysis to determine the relationship between placental DKK1 and lipid accumulation. Maternal plasma and placental NEFA and TG were elevated in OP dams, and offspring of OP dams were smaller than OR. Placental Dkk1 mRNA content was 4-fold lower in OP placentas, and there was a significant increase in β-Catenin accumulation as well as mRNA content of fat transport and TG synthesis enzymes, including Ppar-delta, Fatp1, Fat/Cd36, Lipin1, and Lipin3. There was significant lipid accumulation within the decidual zones in OP but not OR placentas, and the thickness of the decidual and junctional zones was significantly smaller in OP than OR placentas. Overexpression of DKK1 in JEG3 cells decreased lipid accumulation and the mRNA content of PPAR-Delta, FATP1, FAT/CD36, LIPIN1, and LIPIN3. Our results indicate that Dkk1 may be regulating placental lipid metabolism through Wnt-mediated mechanisms. Additionally, recent studies have suggested that maternal obesity may also program early development of non-alcoholic fatty liver disease (NAFLD), rates of which have correlated with the increase in the obesity epidemic. In the current study, livers of OP offspring had significantly increased TG content (P<0.05) and lipid accumulation when compared to offspring of OR dams. Additionally, hepatic Dkk1 mRNA content was significantly decreased in OP livers when compared to OR (P<0.05), and treating H4IIECR rat hepatocyte cells with NEFA showed that Dkk1 mRNA was also decreased in NEFA-treated cells (P<0.05) that also had lipid accumulation. Chromatin Immunoprecipitation (ChIP) analysis of the Dkk1 promoter in fetal livers showed a pattern of histone modifications associated with decreased gene transcription in OP offspring, which agrees with our gene expression data. These results demonstrate that the hepatic Dkk1 gene is epigenetically regulated via histone modification in neonatal offspring in the current model of gestational obesity, and future studies will be needed to determine whether these changes contribute to excessive hepatic lipid accumulation in offspring of obese dams.

Síndrome Alcoólica Fetal: conhecimentos e perceções dos professores do ensino regular

Pós Graduação em Educação Especial - Domínio Cognitivo Motor

Deteção/quantificação de eritrócitos com hemoglobina fetal na gravidez e pós parto utilizando um analisador hematológico

Introdução: As células F(CF) são eritrócitos contendo hemoglobina (Hb) F e outros tipos de hemoglobina. São encontradas em indivíduos de todas as idades, ao contrário dos eritrócitos fetais, só encontrados em fetos e recém-nascidos. Nestes eritrócitos fetais, a Hb F é o tipo dominante de Hb. Estudos publicados indicam que a gravidez pode levar a um aumento progressivo de células com Hb F no sangue materno, devido à presença de CF e/ou de células fetais, estas últimas frequentemente associadas a hemorragia feto–materna (HFM). Objetivo: (1) Determinação da percentagem (%) de células com Hb F em sangue materno, usando um anticorpo Anti-Hb F num analisador hematológico. (2) Quantificação de CF e/ou células fetais na gravidez e pós o parto. (3) Elaboração de algoritmo para a triagem de HFM. Material e Métodos: Estudadas 168 amostras de sangue materno: 29 no 1º trimestre da gravidez (1ºT); 43 no segundo (2º T); 82 no terceiro (3ºT), 14 pós-parto (PP) (amostras entre dia 0 e dia 7, após parto); 32 controlos negativos (Ctl N) com homens adultos saudáveis e 30 controlos positivos (Ctl P), obtidos por mistura de sangue do cordão com sangue do adulto, AB0 compatíveis. Amostras processadas no analisador hematológico Cell-Dyn Sapphire tm, em modo RBC Flow, após ajuste de parâmetros IAS, FL1 e FL3, utilizando um reagente com iodeto de propídio e 2,5 uL de anticorpo monoclonal anti-Hb F FITC. Imagens analisadas pelo software FCS Express V3. Análise estatística com Kruskal-Wallis e teste t-Student (significância estatística p <0,05). Cut-off para HFM obtido pelo valor mínimo, em %, em que se detetam células fetais na amostra Ctl P. Resultados: foram encontradas diferenças estatisticamente significativas na % células com Hb F (P<0,0001) nos grupos estudados. % CF aumenta com a gravidez:1ºT vs Ctl N - p <0,0217 e também durante a gravidez 1ºT Vs 3ºT – P=0,0007. Mesmo depois do PP a % CF está aumentada Ctl N vs PP: - p<0,0001. Valores médios de % CF residuais em adultos saudáveis: 0,53. Maioria das amostras nos diferentes grupos estudados apresenta % CF acima do valor residual (>0,53%): 66% no grupo 1ºT, 83 % no 2º T e 91% no 3º T. Valor cut-off para suspeita de HFM de 1,70% de células com Hb F. Teste preciso (CV+- 4%) para baixas % de células com Hb F. Discussão/Conclusão: Há um aumento células com Hb F durante a gravidez e esse aumento permanece no período PP. Em duas amostras do 3ºT obteve-se % células com Hb F elevada, superior ao cut-off (≥1,70%), sendo detetada uma população de prováveis células fetais. A presença células fetais nestas amostras foi confirmada por citometria de fluxo com Anti-Hb F/ Anti-CA, com subsequente diagnóstico de HFM. Esta metodologia é simples, rápida e não dispendiosa, quando aplicada a um analisador hematológico, representa uma mais-valia no rastreio da HFM. No futuro, pode integrar o protocolo de análises de rotina das grávidas, permitindo detetar as HFM silenciosas, que são a origem de muitas anemias de causa desconhecida em recém-nascidos.

The viability of a tool for fetal health monitoring

Health monitoring has become widespread these past few years. Such applications include from exercise, food intake and weight watching, to specific scenarios like monitoring people who suffer from chronic diseases. More and more we see the need to also monitor the health of new-born babies and even fetuses. Congenital Heart Defects (CHDs) are the main cause of deaths among babies and doctors do not know most of these defects. Hence, there is a need to study what causes these anomalies, and by monitoring the fetus daily there will be a better chance of identifying the defects in earlier stages. By analyzing the data collected, doctors can find patterns and come up with solutions, thus saving peoples’ lives. In many countries, the most common fetal monitor is the ultrasound and the use of it is regulated. In Sweden for normal pregnancies, there is only one ultrasound scan during the pregnancy period. There is no great evidence that ultrasound can harm the fetus, but many doctors suggest to use it as little as possible. Therefore, there is a demand for a new non-ultrasound device that can be as accurate, or even better, on detecting the FHR and not harming the baby. The problems that are discussed in this thesis include how can accurate fetus health be monitored non-invasively at home and how could a fetus health monitoring system for home use be designed. The first part of the research investigates different technologies that are currently being used on fetal monitoring, and techniques and parameters to monitor the fetus. The second part is a qualitative study held in Sweden between April and May 2016. The data for the qualitative study was collected through interviews with 21 people, 10 mothers/mothers-to-be and 11 obstetricians/gynecologists/midwives. The questions were related to the Swedish pregnancy protocol, the use of technology in medicine and in particular during the pregnancy process, and the use of an ECG based monitoring device. The results show that there is still room for improvements on the algorithms to extract the fetal ECG and the survey was very helpful in understanding the need for a fetal home monitor. Parents are open to new technologies especially if it doesn't affect the baby's growth. Doctors are open to use ECG as a great alternative to ultrasound; on the other hand, midwives are happy with the current system. The remote monitoring feature is very desirable to everyone, if such system will be used in the future.

Desarrollo y dinámica de los folículos ováricos desde la etapa fetal hasta la prepuberal en bovinos

Resumen Esta revisión compila el desarrollo y la dinámica folicular con una perspectiva histológica y de biología del desarrollo, desde la etapa fetal hasta prepuberal en bovinos (Bos taurus). Se trata de un proceso complejo y coordinado, con cambios fisiológicos y morfológicos que intervienen en la diferenciación y el desarrollo ovocitario. Los folículos son la unidad fundamental del ovario y comandan los procesos reproductivos y las fases del ciclo estral, con dos funciones fundamentales, la producción de hormonas y de ovocitos. La formación de folículos primordiales se conoce como el proceso de ensamblaje folicular, en el cual “nidos” de ovocitos completan su proliferación mitótica y comienzan la meiosis, mientras que muchos de estos folículos se vuelven apoptóticos, favoreciendo la desorganización de los nidos. El proceso de formación de folículos primordiales comienza durante el desarrollo fetal en bovinos, aproximadamente a los 80 días de gestación. Existe gran variación individual en el número de folículos, existiendo un pool de reserva ya al nacimiento. De unos 2.700.000 folículos primordiales constituidos al final del ensamblaje folicular, 90% degeneran, quedando al nacimiento aproximadamente 135.000 células germinales, las cuales declinan rápidamente hasta la pubertad y luego de ella. Los folículos primordiales se desarrollan a folículos primario, secundario, antral y preovularorio (en hembras adultas). Estas transiciones implican cambios citológicos, histológicos y morfológicos que son descritos. En terneras, igual que en animales adultos, ocurren ondas de crecimiento folicular, que aparecen a la segunda semana de vida. En la pubertad se observan importantes cambios endócrinos y ováricos y es influida por múltiples factores. El desarrollo folicular está íntimamente vinculado al futuro desempeño reproductivo de las hembras, por lo cual su conocimiento es fundamental para un adecuado manejo reproductivo de la hembra bovina y para ampliar las investigaciones vinculadas a esta disciplina

Zika virus damages the human placental barrier and presents marked fetal neurotropism

An unusually high incidence of microcephaly in newborns has recently been observed in Brazil. There is a temporal association between the increase in cases of microcephaly and the Zika virus (ZIKV) epidemic. Viral RNA has been detected in amniotic fluid samples, placental tissues and newborn and fetal brain tissues. However, much remains to be determined concerning the association between ZIKV infection and fetal malformations. In this study, we provide evidence of the transplacental transmission of ZIKV through the detection of viral proteins and viral RNA in placental tissue samples from expectant mothers infected at different stages of gestation. We observed chronic placentitis (TORCH type) with viral protein detection by immunohistochemistry in Hofbauer cells and some histiocytes in the intervillous spaces. We also demonstrated the neurotropism of the virus via the detection of viral proteins in glial cells and in some endothelial cells and the observation of scattered foci of microcalcifications in the brain tissues. Lesions were mainly located in the white matter. ZIKV RNA was also detected in these tissues by real-time-polymerase chain reaction. We believe that these findings will contribute to the body of knowledge of the mechanisms of ZIKV transmission, interactions between the virus and host cells and viral tropism.

Valor predictivo del perfil biofísico fetal en gestantes a término en relación con las condiciones neonatales detectadas por el APGAR. Hospital Regional Vicente Corral Moscoso, enero-septiembre 2005

Se justifica el estudio como un aporte importante que permite en base al trabajo en equipo de a profesionales en imágenes, pediatras y obstetras, responsables del binomio madre-hijo, tener un método útil o de exploración peri.natal que ha permitido evaluar una serie de actividades biofísicas que ayudan en la toma de decisiones certeras en la conducción del parto y/o culminación del embarazo, que ayudan a predecir varias complicaciones entre las más importantes el sufrimiento fetal agudo, consecuencia de la hipoxia que muchas de las veces ha terminado en la muerte fetal. Por lo que se hace necesario determinar el valor predictivo de la prueba del Perfil Biofísico Fetal en gestantes a término en relación con las condiciones neonatales valoradas por el Test de Pagar al momento del nacimiento. Conclusiones: en relación a la prueba de perfil biofísico fetal se encontró que la sensibilidad de la prueba es de 62.96, la especificidad de 93.45, el valor predictivo positivo fue del 48.5, el valor predictivo negativo 96.45, los índices de verdaderos positivos de 48.47, de falsos positivos de 51.43, de falsos negativos 3.75y de verdaderos negativos 96.25

The Role of Mucosal Defense in Intestinal Injury of Infants With Fetal Growth Retardation

Background: Infants with fetal growth retardation (FGR) are prone to intestinal disorders. Objectives: Aim of the study was to determine the role of mucosal defense ability in formation of gut injury in infants with FGR. Materials and Methods: 44 premature infants who were admitted to the Neonatal Intensive Care Unit were divided into two groups: 20 infants with FGR (FGR group) and 24 appropriate-for-gestational age newborns (AGA group). Control group consisted of 22 premature infants who were delivered after uncomplicated pregnancy. Gut barrier function was evaluated by detecting serum intestinal trefoil factor (ITF) and intestinal fatty acid binding protein (IFABP). The level of serum IFABP and ITF was measured by using ELISA method. Results: FGR group showed significantly higher ITF concentration than AGA group on the first days of life (P ˂ 0.01). High level of ITF in the FGR group significantly declines up to 7th - 10th day of life (P ˂ 0.01). This reduction was accompanied by increase of IFABP which is a marker of ischemic intestinal mucosal injury. Correlation analyses showed that ITF had a negative correlation with IFABP. Conclusions: Infants with fetal growth retardation are characterized by a high level of ITF on the first days of life. This protects intestinal mucosa under hypoxic conditions. Its subsequent decline accompanied by an increase of IFABP reflects the depletion of Goblet cells to secret ITF causing damage to the integrity of intestinal mucosal barrier.

Mal nutrición materna, parto y bienestar fetal. Hospital Vicente Corral Moscoso. Abril-Julio 2007

Determinar la prevalencia de malnutrición materna al parto, relacionar con el tipo de parto y la condición del recién nacido. Embarazadas al parto, departamento de obstetricia, Hospital Vicente Corral Moscoso, Abril - Julio 2007, Muestra: 650 parturientas. Al ingreso se peso y tomo la talla, clasificó según el nomograma de Rosso y Mardones. Se valoró el tipo de parto. Se clasificó según peso, edad gestacional y según riesgo de mortalidad neonal (Bataglia y Lunchenko), APGAR a los cinco minutos y el destino del recien nacido. La prevalencia de malnutrición es 68 por ciento. 27.4 por ciento son adolescentes. Parto vaginal 67.8 por ciento, cesárea el 32.2 por ciento. El recién nacido (RN) 70.9 por ciento es adecuado, 20.9 por ciento es pequeño y el 8.2 por ciento es grande para la edad gestacional. El 98.5 por ciento son vivos. El APGAR mayor 7 a los cinco minutos 93.3 por ciento. El 87 por ciento de RN el destino es su madre, 10.6 por ciento a neonatología. El peso bajo de la madre se asocia con el recién nacido pequeño (OR=1,695). El peso bajo de la madre es protector para el RN sea grande (OR=0,313; el sobrepeso (OR=2,098), la obesidad (OR=2,941) es asociación con el RN sea grande. La obesidad materna es asociación con el al APGAR menor 7 (OR=2,007). El parto por cesárea es asociación con el RN pequeño (OR=2,011), el destino a neonatología (OR=2,368) y APGAR menor 7 (OR=2,314)

Densidad óptica en embarazadas normales y madurez pulmonar fetal. Estudio en el líquido amniótico. Hospital Vicente Corral Moscoso. Cuenca -Ecuador 1999

Se realiza un estudio de tipo clínico-descriptivo con el objeto de determinar la presencia de pigmentos bilirrubinoides y la madurez pulmonar, comprado el Test de Clements con la edad gestacional y el estado clínico del recién nacido. Se tomó a cincuenta pacientes embarazadas entre 37 y 41 semanas de gestación, sin factores de riesgo y sus recién nacidos, atendidos mediante cesárea o parto vaginal, en el Centro Obstétrico del Hospital Vicente Corral Moscoso de la ciudad de Cuenca en 1999. Se realizaron las determinaciones basados en los parámetros recomendados a nivel internacional concluyendo que: 1 En las embarazadas a término de la gestación, la determinación de la presencia de pigmentos bilirrubinoides en líquido ammniótico corresponde a la zona 1 de la gráfica de Liley, interpretada como exenta de riesgo para los recién nacidos, de acuerdo con los parámetros que se registran en la literatura médica actual. 2. La prueba de Clements positiva guarda estrecha relación con la madurez del recién nacido expresada por Capurro y con una adecuada función respiratoria expresada por un puntaje normal de Silverman en todos los casos. 3. En caso de usarse la prueba de Clements, es la prueba de un tubo, suficiente para valorar la madurez pulmonar por su alta confiabilidad y bajo costo. 4. Por lo tanto los autores recomendamos que se incluya la aplicación de estas dos pruebas, que son de fácil realización, como parte del protocolo de atención materno-infantil en el Centro Obstétrico del Hospital Vicente Corral Moscoso

Indicadores ecográficos de patología fetal del tercer trimestre de gestación: Hospital Vicente Corral Moscoso, febrero-julio del 2005

Las experiencias prenatales y postnatales de este grupo de gestantes investigadas, tanto por ultrasonido como por valoración clínica pediátrica; ha sido de mucho interés. El objetivo consiste en detectar mediante signos ecográficos la presencia de patología fetal; utilizando un ecógrafo marca Sonoace 8000 EX con transductor convexo 3,5 MHZ, puesto que se han comparado punto por punto la investigación con la experiencia de grupos similares de otras investigaciones, efectuándose por lo tanto una correlación ecográfica y clínica en toda mujer embarazada que cursa el tercer trimestre de gestación, obteniéndose de las 355 pacientes estudiadas, 28 casos positivos para patología fetal por ultrasonido, de los cuales se comprobó clínicamente al momento del nacimiento 20 casos, lo que permite indica que se puede diagnosticar un 71.4cuadros patológicos sonoembrionarios; que en comparación con grupos similares de investigación, es significativo. En el grupo de personas investigadas hubo también falsos positivos para ultrasonido tal es el caso de 6 (20) en patología craneal, 1 (10) en polimalformados y 1 5en patología toráxica; los demás coincidieron en su diagnóstico prenatal con el postnatal. De esta manera señalamos que en la presente investigación las pruebas diagnosticas son tambien significativas y acordes a las investigaciones que trae la literatura mundial, en donde se reportan sensibilizadas bajas, que van desde 24.5(González 1999), hasta 73.7(estudio mundial Eurofetus), con especialidades altas sobre los 90; el grupo investigado posee una sensibilidad del 70y una especificidad del 95.82que indica que se puede captar en el 70a los verdaderos positivos y en el 95a los verdaderos negativos con una probabilidad de padecer la enfermedad se si obtiene un resultado positivo en la prueba que es de un 50, con lo que se concluye e insiste en la importancia que posee el manejo interdisciplinario y la aplicación de esta modalidad en toda gestante