Differential expression profile of miRNAs in amniotic fluid exosomes from fetuses with Down syndrome / 南方医科大学学报

OBJECTIVE@#To investigate the role of miRNAs in amniotic fluid exosomes in growth and development of fetuses with Down syndrome (DS).@*METHODS@#Amniotic fluid were collected from 20 fetuses with DS and 20 normal fetuses (control) to extract amniotic exosome miRNA. MicroRNA sequencing technique was used to identify the differentially expressed miRNAs between the two groups, for which gene ontology (GO) and pathway analysis was performed. Three differentially expressed miRNAs with the strongest correlation with DS phenotype were selected for qPCR verification. Dual luciferase reporter assay was used to verify the activity of let-7d-5p for targeted regulation of BACH1.@*RESULTS@#We identified 15 differentially expressed miRNAs in DS as compared with the control group, among which 7 miRNAs were up-regulated and 8 were down-regulated. Target gene prediction results showed that the differentially expressed miRNAs targeted 17 DS-related genes. GO analysis revealed that the main functions of the target genes involved protein binding, protein transport, ATP binding, transferase activity and synapses. Pathway analysis revealed that the functional pathways were closely related with the development of the nervous system. qPCR results showed that the expression levels of miR-140-3p and let-7d-5p were significantly lower in DS group than in the control group (P < 0.05), as was consistent with miRNA sequencing results; the expression level of miR-4512 was significantly higher in DS group than in control group (P < 0.05), which was contrary to miRNA sequencing results. The results of double luciferase reporter gene assay confirmed that let-7d-5p was capable of targeted regulation of BACH1 expression.@*CONCLUSION@#Let-7d-5p in amniotic fluid exosomes may promote oxidative stress events in the brain of fetuses with DS by regulating BACH1 expression.

Aspectos imunogenéticos do parto pré-termo espontâneo / Immunogenetic aspects in spontaneous preterm birth

O parto pré-termo é uma das grandes intercorrências obstétricas, sendo a maior causa de morbidade e mortalidade perinatal. Dentre os diferentes fatores envolvidos no seu desencadeamento, a infecção intra-amniótica parece representar um papel central. As infecções desencadeiam resposta inflamatória nos tecidos materno e fetal, mediada pela produção de citocinas inflamatórias. As citocinas induzem a liberação de prostaglandinas, aumentando a contratilidade uterina, favorecendo a rotura das membranas fetais, a modificação e dilatação da cérvice e, finalmente, o parto pré-termo. A síntese de citocinas depende de controle genético. Diversos polimorfismos relacionados a genes humanos codificadores de citocinas são reconhecidos e associados a fenótipos de alta, média e baixa produção desses fatores. Assim sendo, a relação entre determinados genótipos e a ocorrência e/ou características de diferentes patologias tem sido investigada. Este tipo de abordagem pode contribuir para o conhecimento da patogenia, permitindo o reconhecimento de parâmetros preditivos e a definição de novas estratégias terapêuticas.

Preterm birth is a major obstetric incident and one of the main causes of perinatal mortality and morbidity. Among the different factors involved in its unleashing, intra-amniotic infection seems to play a central role. The infections unleash inflammatory response in both maternal and fetal tissues, mediated by the production of inflammatory cytokines. They also lead to liberation of prostaglandin, which increases myometrial contractility, favoring the rupture of fetal membrane, alteration and dilation of the cervix and, finally, preterm birth. The production of cytokines depends on genetic control. Many polymorphisms related to human genes that codify cytokines are recognized and associated with phenotypes of high, medium and low production of such factors. Thus, the relation between certain genotypes and the occurrence and/or characteristics of several pathologies has been the focus of investigations. This approach may contribute with a better understanding of the pathogenesis, allowing the identification of predictive parameters and the establishment of new intervention strategies.

Maternal Serum and Amniotic Fluid Inhibin A Levels in Women who Subsequently Develop Severe Preeclampsia

The purpose of this study was to evaluate whether maternal serum (MS) and amniotic fluid (AF) inhibin A levels are elevated in patients who subsequently develop severe preecalmpsia, and to investigate the correlation between MS and AF inhibin A levels in the second trimester. The study included 40 patients who subsequently developed severe preecalmpsia and 80 normal pregnant women. Inhibin A levels in MS and AF were measured with enzyme-linked immunosorbent assay (ELISA). The MS and AF inhibin A levels in patients who developed severe preeclampsia were significantly higher than those in the control group (both for p<0.001). There was a positive correlation between MS and AF inhibin A levels in patients who developed severe preeclampsia (r=0.397, p=0.011), but not in the control group (r=0.185, p=0.126). The best cutoff values of MS and AF inhibin A levels for the prediction of severe preeclampsia were 427 pg/mL and 599 pg/mL, respectively; the estimated ORs that were associated with these cut-off values were 9.95 (95% CI 3.8-25.9, p<0.001) and 6.0 (95% CI 2.3-15.8, p<0.001). An elevated level of inhibin A in MS and AF at the time of second trimester amniocentesis may be a risk factor for the subsequent development of severe preeclampsia.

Avaliação prospectiva do índice de líquido amniótico em gestações normais e complicadas / Prospective evaluation of amniotic fluid index in normal and high risk pregnancies

OBJETIVO: Analisar, prospectivamente, o índice de líquido amniótico em gestantes de baixo risco que apresentaram ou não complicações da gestação e do período perinatal. MATERIAIS E MÉTODOS: Foi estudado, de modo prospectivo, o índice de líquido amniótico de 45 primigestas sem doenças no momento da inclusão no estudo. O índice foi avaliado em quatro períodos da gestação - 18-20, 24-26, 28-30 e 34-36 semanas - e seus valores foram correlacionados com o surgimento de complicações gestacionais e perinatais, Doppler das artérias uterinas, idade gestacional ao parto, vias de parto e o peso dos recém-nascidos. RESULTADOS: O índice de líquido amniótico não mostrou diferença estatística entre as pacientes com gestações normais e complicadas em nenhum dos quatro períodos estudados. Também não mostrou associação com a idade gestacional ao parto, as vias de parto e o peso dos recém-nascidos. Observou-se que a média do índice de líquido amniótico entre 28-30 semanas é mais elevada (p = 0,004) nas pacientes com incisura bilateral do que nas com Doppler normal. CONCLUSÃO: A avaliação prospectiva do índice de líquido amniótico em gestações de baixo risco parece não ser um bom preditor de complicações gestacionais ou perinatais.

OBJECTIVE: To prospectively analyze the amniotic fluid index of low-risk pregnant women who presented or not complications during pregnancy and perinatal period. MATERIALS AND METHODS: The amniotic fluid index was prospectively studied in 45 first pregnancy women with no diseases at study entry. The amniotic fluid index was determined at four time points during pregnancy - 18–20, 24–26, 28–30 and 34–36 weeks - and the values were correlated with the appearance of pregnancy and perinatal complications, uterine artery Doppler findings, gestational age at delivery, route of delivery, and newborn weight. RESULTS: Amniotic fluid index was not significantly different among patients with normal and high risk pregnancies at any of the four time points studied. Likewise, there was no association with gestational age at delivery, route of delivery or newborn weight. The mean amniotic fluid index at 28–30 weeks was higher (p = 0.004) in patients with bilateral incisure than in patients with normal Doppler. CONCLUSION: Prospective evaluation of amniotic fluid index in low-risk pregnancies does not seem to be a good predictor of complications during pregnancy or perinatal period.

Ultrasonographic determination of amniotic fluid volume in normal pregnancy.

Ultrasonographic assessment of amniotic fluid has important implications because documentation of abnormalities of amniotic fluid volume may provide valuable information to enhance fetal health assessment. But in our country no study was conducted to establish normal values of amniotic fluid volume. So, this study was designed to find out normal values of amniotic fluid volume at various stages of gestation. The study was conducted in the department of Radiology and Imaging, BIRDEM in collaboration with the department of Obstetrics and Gynaecology, BIRDEM from 1(st) July, 2001 to 30(th) June, 2002. One hundred and twenty patients of normal pregnancy between 15 to 38 weeks' gestation were scanned by ultrasonography and their amniotic fluid volume was measured. For each gestational age between 15 and 38 weeks, 5 patients were recruited, and only one examination was done per pregnancy. Currently, the three semi-quantitative methods of assessing amniotic fluid volume include the amniotic fluid index (AFI), single deepest pocket (SDP) and two diameter pocket (TDP). Amniotic fluid volume of the study population was measured by the three methods. The values were plotted against each week in tabulated form. Thus the normal range of amniotic fluid volume in three methods (AFI, SDP, TDP) across gestational age in normal pregnancies was established. The mean value of amniotic fluid volume in AFI, SDP and TDP were 13.224 cm, 4.679 cm and 19.252 cm(2) respectively. AFI and SDP showed amniotic fluid volume increased with increasing gestational age up to early third trimester, but volume decreased thereafter. The prevalence of false positive result for oligohydramnios was 0.83% for AFI, 0% for SDP and 23% for TDP. The prevalence of false positive result for hydramnios was 1.66% for AFI, 0.83% for SDP and 0% for TDP. Thus the normal values of amniotic fluid volume for each of the three ultrasonographic techniques in the perspective of our country were obtained from the study. As the study was conducted with a limited number of patients, further study may be carried out with a large number of observations to reevaluate the results of the study.

Newly Developed Mecometer Method for Objective Assessment of Meconium Content

We developed a new method for an objective assessment of the meconium content in amniotic fluid. By establishing a standard scale through a serial dilution of a known amount of meconium into the amniotic fluid, we developed a new method 'mecometer 'that can objectively measure the meconium content in meconium-stained amniotic fluid samples. The objectivity and reliability of this mecometer were verified by 300 student volunteers. At least 70% of the volunteers could objectively measure and digitally describe the meconium content in meconium-stained amniotic fluid samples. We believe our newly developed mecometer is a very simple, reliable, and portable method, not requiring any instruments.

Amniotic fluid interleukin-6 and the risk of Early-Onset sepsis among preterm infants

Background. High concentrations of interleukin-6 (IL-6) have been demonstrated in amniotic fluid (AF) from women with intra-amniotic infection. Recent studies have reported that IL-6 levels in AF were related to a increase in neonatal morbidity; moreover, higher Il-6 plasma levels have been observed in neonates with sepsis. Methods. A cohort study was carried out at the National Institute of Perinatology in Mexico City. Inclusion criteria were the following: 1) preterm singleton pregnancy; 2) intact membranes at time of enrollment, and 3) written informed consent. Women with other complications of pregnancy were excluded. Newborn sepsis during the first 72 h was defined as early-onset sepsis. Amniotic fluid was obtained at the moment of delivery. Amniotic fluid Il-6 (Af IL-6) was determined by enzyme-linked immunoassays. Results. Ninety-three women met the criteria for enrolment in the study and 31 (33 percent) of their newborns had early-onset neonatal sepsis. The mean AF IL-6 in mothers of septic newborns was 5779 ñ 2804 pg/ml compared to 729 ñ 382 pg/ml in mothers with noninfected neonates (p<0.001). AF IL-6 concentrations higher than 1250 pg/ml were significantly associated with early-onset sepsis (OR 33.3: 95 percent CI 9.4-117.3) (p< 0.001). Gestational age under 32 weeks was also associated with neonatal sepsis (OR 2.56; 95 percent CI 1.2-9) (p = 0.002). Women whose infants developed neonatal sepsis had a higher frequency of clinical choriamnionitis (p = 0.02). Conclusions. IL-6 determination in AF may be a useful indicator to identifity neonates with higher risk of in utero bacterial infection

Valor predictivo de las pruebas de madurez pulmonar fetal en la diabética embarazada / Predictive value of fetal lung maturity tests in diabetic pregnant women

El síndrome de dificultad respiratoria (SDR) es una morbilidad neonatal común en el hijo de madre diabética. Sin embargo, en la literatura existe controversia sobre la aparición de madurez pulmunar en estos infantes, careciendo varios estudios de un verdadero grupo testigo. Se comparó la confiabilidad predictiva de perfil de madurez pulmonar fetal (MPF): densidad óptica (DO), lecitina esfingomielina (L/E), y fosfatil glicerol (F/G), en un grupo de pacientes diabéticos y en un grupo de embarazadas sanas. Se estudiaron prospectivamente las pruebas de MPF, obtenidas por aminocentesis a partir de las 37 semanas, en 47 pacientes: 20 diabéticas (grupo de estudio) y 27 embarazadas normales (testigo). Todos los embarazos se interrumpieron por cesárea. Se excluyeron neonatos con malformaciones mayores, madres complicadas con hipertensión, sospecha de RCIU y diabetes tipo I. Los valores de las pruebas de MPF, en ambos grupos, se correlacionaron con presencia de SDR. Tratamiento estadístico: sensibilidad, especificidad, valores predictivos (+) u (-), prueba exacta de Fisher para asociación de cada prueba con SDR y t de Student. Cinco de 49 neonatos tuvieron SDR (incidencia global: 10.2 por ciento), 4 de estos 5 neonatos con SDR, pertenecieron al grupo de diabéticas, incidencia de 20 por ciento, en contraste con un solo caso de 29 neonatos controles (3.7 por ciento), esta diferencia tuvo alta significancia estadística p< 0.0001. Entre estos casos con SDR hubo 4 con prueba de MPF falsamente positivas: 3 ocurrieron en la relación L/E del grupo de diabéticas (3/20 = 15 por ciento) y la 4ta.falsa + en la DO de este mismo grupo. No hubo ningún falso + para el FG (especificidad 100 por ciento), en ambos grupos. La mayor sensibilidad en el grupo de estudio correspondió al FG (39 por ciento), así como 100 por ciento de valor predictivo + (todos los neonatos con SDR con pruebas inmaduras). Las 3 pruebas de madurez en el grupo testigo tuvieron 100 por ciento de especificidad. Este estudio confirmó que la presencia de FG en líquido amniótico elimina totalmente el riesgo de SDR en el hijo de madre diabética y que su ausencia constituye un marcador bioquímico de SDR. La relación L/E en la embarazada diabética puede tener resultados falsos + en un porcentaje no despreciable 15 por ciento o principalmente cuando hay descontrol metabólico severo

Repercussao neonatal do aspecto do liquido amniotico e das caracteristicas dos batimentos cardiacos fetais durante o trabalho de parto / Neonatal repercussion of the aspect of the amniotic fluid and the fetal heart rate characteristics during labor

Foram analisados 2741 partos ocorridos na Maternidade do Departamento de Tocoginecologia da UNICAMP, para estudo da repercussao neonatal do aspecto do liquido amniotico e das caracteristicas dos batimentos cardiacos fetais durante o trabalho de parto. O aspecto do liquido amniotico foi distribuido em cinco grupos: claro, ligeiramente tingido de meconio, tingido de meconio, com meconio espesso e com meconio antigo. A presenca de batimentos cardiacos fetais normais e anormais foi considerada em cada grupo, e as condicoes neonatais avaliadas pelos indices de Apgar no primeiro e quinto minuto. Concluiu-se que os resultados neonatais sao semelhantes nos casos com liquido amniotico claro, ligeiramente tingido e tingido de meconio, enquanto partos que evoluem com meconio espesso e antigo aumentam significativamente a incidencia de recem-nascidos deprimidos....

Padronizaçäo das concentraçöes de esteróides no líquido amniótico: II. Testosterona, sulfato de deidroepiandrosterona e cortisol / Standardization of steroid concentrations in the amniotic fluid: II. Testosteroni, dehydroepiandrosterone sulfate and cortisol

As dosagens de testosterona, sulfato de deidroepiandrosterona (SDHEA) e cortisol no líquido amniótico (LA) permitem avaliar as funçöes das adrenais e das gônadas fetais. Estudamos LAs de 41 gestaçöes, obtidos por amniocentese no segundo trimestre da gestaçäo. Trinta e sete destas gestaçöes (das quais quatro de risco para hiperplasia adrenal congênita por deficiência da 21-hidroxilase) resultaram em fetos normais, três foram anencéfalos e um, portador de deficiência da 21-hidroxilase. Os esteróides foram dosados utilizando-se radioimunoensaio previamente descritos para dosagens séricas. Valores médios (ñ DP) destes esteróides no LA de gestaçöes com fetos normais foram testosterona: 76 ñ 34 ng/del; SDHEA: 22 ñ 9 ng/ml; cortisol 19 ñ 8 ng/ml. LAs de gestaçöes que resultaram em anencéfalos apresentaram concentraçöes de SDGEA e cortisol significativamente mais baixas em relaçäo aos normais. Testosterona foi mais baixa em fetos femininos quando comparados aos masculinos normais e discretamente mais elevada que o normal no feto feminino afetado.Näo houve diferença no valor destes esteróides ao compararmos LAs de fetos de risco para a deficiência da 21-hidroxilase com os normais. Assim, a dosagem destes esteróides näo contribui de modo definitivo para o diagnóstcio de defeitos de síntese adrenal ou de anencefalia. Também näo permitem definir com segurança o sexo fetal

Actividad amilásica y relación lecitina-esfingomielina en líquido amniótico como parámetros de madurez pulmonar fetal

La medida de la actividad amilásica en líquidos amnióticos (L.A.) ha sido propuesta como prueba "screening" para determinar madurez fetal, Nosotros hemos reunido y examinado 272 muestras de L.A. para dosar amilasa y evaluar la relación lecitina-esfingomielina. La amilasa del L.A. mostró una correlación pobre con respecto a la relación lecitina-esfingomielina (r: 0.240). El análise de la amilasa como prueba "screening" para determinar la necesidad de evaluar la relación lecitina-esfingomielina mostró una sensibilidad del 54% y una especificidad del 88%. Se estudiaron tres grupos de gestantes: Un grupo con amilasa inferior a 200 U/L, un grupo intermedio con valores entre 200 y 300 U/L y un grupo de pacientes con amilasa superior a 300 U/L. Solamente el 46% del grupo con baja amilasa, tenía relación lecitina-esfingomielina inmadura. El grupo con cifras de amilasas altas, mostró la mejor correlación entre amilasa y relación lecitina-esfingomielina, pero un 12% de estas muestras dieron lugar a una relación lecitina-esfingomielina inmadura o "borderline". Se concluye que la amilasa del L.A. no debe ser usada como prueba "screening" para determinar la necesidad de recurrir a la evaluación de la relación lecitina-esfingomielina

A quantitative study of human amniotic fluid fibronectin in the second trimester of pregnancy

Foi determinada por eletroimunoensaio a concentraçäo de fibronectina em 60 amostras de líquido aminiótico obtidas de gravidezes normais no segundo trimestre. A concentraçäo média de fribronectina foi de 142 + ou - micron-grama/ml (média + ou - erro padräo), correspondendo a 3,1% da proteína total do líquido amniótico. A concentraçäo de fibronectina elevou-se significamente entre a 14ª e a 24ª semanas de gravidez e apresentou uma correlaçäo negativa significativa com os níveis de alfa-fetoproteína no líquido amniótico; esta correlaçäo parecia ser espúria e devida a um efeito da idade gestacional sobre ambas variáveis. A concentraçäo de fibronectina estava dentro de limites normais em duas amostras de fetos patológicos, sendo um com síndrome de Down (trissomia 21) e o outro com anencefalia

Sales biliares (colilglicina) en líquido amniótico y en sangre de cordón umbilical en pacientes con colestasia intrahepática del embarazo (CIE) / Biliary salts (cholylglicine) in amniotic fluid and blood of umbilical cord in patients with intrahepatic cholestasia of pregnancy

Se diseña un estudio para conocer el comportamiento de la colilglicina, CG, en sangre materna, sangre total de cordón umbilical y en líquido amniótico, en embarazadas sanas y con colestasia intrahepática del embarazo, CIE, en muestras tomadas simultáneamente. A las pacientes seleccionadas y sin otra patología materna o fetal se las dividió en dos grupos. El grupo control está formado por 14 embarazadas sanas, y el grupo con CIE, por ocho pacientes. Se hizo un diagnóstico de CIE por el cuadro clínico, niveles de CG en ayunas superiores a 0,7 ug/ml., y desaparición del prurito después del parto. Durante la cesárea se obtuvo muestra simultánea de líquido amniótico, sangre total del cordón y sangre venosa materna, y se determino CG mediante enzimo-inmunoensayo ENDAB-cholylglicine. Se encontraron valores promedios de CG algo más elevados en sangre de cordón que en sangre materna en ambos grupos, pero sin diferencias significativas: grupo control, X=0,6 más menos 0,7 ug/ml vs. X=0,14 más menos 0,1 ug/ml; grupo con CIE= 2,3 más menos 1,5 ug/ml vs. X=1,6 más menos 0,6 ug/ml. Al comparar por separado cada conjunto madre-hijo se encontró que los valores CG fetal pueden ser diferentes a los de su madre (3 casos), o presentar una correlación estadísticamente significativa: después de eliminar valares extremos, tres muestras donde Z > 1,65 al aplicar prueba de covarianza. Grupo control, R=0,621, p < 0,02; grupo CIE, R=0,958 p < 0,01. Los niveles CG en sangre de cordón en los hijos de madres CIE son tres veces mayores que en los hijos del grupo control. Los valores de CG en líquido amniótico presentaron amplias variaciones en el grupo con CIE.

Estudio crítico de la actividad tromboplástica del líquido amniótico en diversas semanas gestacionales / Critical study of the tromboplastic activity of the amniotic fluid in different gestational weeks

Se determinó la actividad tromboplástica del LA en setenta y ocho embarazos normales y patológicos por el método de Quick, en una etapa, modificado por Yaffe, para la medida del tiempo de protombina. Se comprobó el aumento de la actividad tromboplástica con el progreso de la gestación, siendo el coeficiente de correlación r = -0,76. No se observaron diferencias entre los valores de actividad tromboplástica entre embarazos normales y patológicos. En tres de cuatro casos de embarazos postérmino, la actividad tromboplástica del LA fue inferior a 45 s. Nuestros resultados corroboraron los enunciados por Yaffe y difieren de los de Phillips y Davidson

El aumento de la actividad tromboplástica del líquido aminiótico como prueba de madurez fetal (test de aceleración de la coagulación / Tromboplastin increased activity of amniotic fluid as fetal maturity test (blood coagulation acceleration test)

Se presentan los resultados del estudio del líquido amniótico de 50 embarazadas hecho con la finalidad de evaluar la actividad tromboplastínica del mismo con miras a encontrar un método rápido, seguro y de bajo costo pero la medición de la madurez fetal. Además del TAC se analizaron otros parámetros: creatinina, peso fetal, aspecto del líquido y semanas de gestación, ésta última calculada retrospectivamente a partir de la edad gestacional del recién nacido determinada mediane el examen neonatal practicado por pediatras del hospital. Se encontró que los valores del TAC descienden a medida que progresa el embarazo, pero los cifras obtenidas en este estudio son mucho más bajas que las dadas por otros autores. La cifra promedio del TAC fue de 77 segundos pero los embarazos de menos de 36 semanas. No observamos buena correlación entre los pesos fetales y los valores de creatinina con el TAC. Consideramos que el aspecto del líquido amniótico continúa siendo una ayuda importante como una prueba más en la estimación de la madurez fetal: en el 77,4% de los casos hubo concordancia entre el TAC el aspecto del líquido

Histologic explaination for reduced fetal urine production and for reduced amount of amniotic fluid in complicated pregnancy.

Reduction in the production of fetal urine and amniotic fluid has been recognised to have resulted from increased tubular mass in the kidneys of sub-acutely or chronically stressed fetuses and from reduced arteriolar luman size in kidney of acutely stressed fetuses.