RESUMEN
OBJECTIVE: Human mesenchymal stem cells (hMSCs) are a promising source for regenerative medicine, especially mesodermal lineages. Clinical applications require an understanding of the mechanisms for transcriptional control to maintain the desired cell type. The aim of this study was to identify novel markers for differentiation of hMSCs into bone or cartilage with the use of Kartogenin, by RNA analysis using microarray technology, and explore the role of RhoA-Rho associated protein kinase (ROCK) inhibition in these. METHODS: Commercial human bone marrow derived primary mesenchymal stem cells were purchased from ATCC. Cells were differentiated in vitro in 2-dimensional cultures using Kartogenin as the main cartilage inducer and bone morphogenetic protein 2 for bone differentiation; cells were cultured with and without ROCK inhibitor Y-27632. After 21 days of culture, whole RNA was extracted and analyzed via Affimetrix microarrays. The most significant hits were validated by quantitative polymerase chain reaction. RESULTS: We found a total of 1,757 genes that were either up- or downregulated on differentiation, when compared to P1 hMSC (control) at day 0 of differentiation. Two members of the Serpin superfamily, SERPINA9 and SERPINB2, were significantly upregulated in the cartilage groups, whereas they were unchanged in the bone groups with and without ROCK inhibition. CONCLUSIONS: SERPINA9 and SERPINB2 are novel differentiation markers, and molecular regulator candidates for hMSC lineage commitment toward bone and cartilage, providing a new tool for regenerative medicine. Our study highlights the roles of these 2 genes, with significant upregulation of both in cell cultures stimulated with Kartogenin.
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Antígenos de Diferenciación/genética , Cartílago/citología , Linaje de la Célula/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Células Madre Mesenquimatosas/citología , Proteínas de Neoplasias/metabolismo , Serpinas/metabolismo , Anilidas , Proteína Morfogenética Ósea 2 , Diferenciación Celular/genética , Células Cultivadas , Humanos , Ácidos Ftálicos , ARN/aislamiento & purificación , Regulación hacia Arriba/genéticaRESUMEN
OBJECTIVE: The purpose of this article was to describe our experience with the prenatal diagnosis of CHD in patients referred to our Fetal Cardiology Unit. METHODS: Prospective cohort study of consecutive fetuses referred for advanced fetal echocardiography to our Fetal Cardiology Unit during a 3-year period (September 2015-September 2018). RESULTS: Totally 809 fetuses were evaluated, with 1263 fetal advanced echocardiographies performed. Suspected cardiac abnormality was the most common indication for referral (62.2%). Only 7.3% of patients had known morbidities or risk factors for CHD. Mean gestational age at first examination was 25.6 ± 6.4 weeks. A total of 528 (65.3%) fetuses were found to have a cardiac defect: 40.7% had isolated CHD while 24.6% had associated anomalies. The most common defects found were ventricular septal defects (20.3%), followed by conotruncal defects (9.7%), hypoplastic left heart syndrome (9.3%), fetal arrhythmias (8.9%), and venous anomalies (8.7%). 31.6% presented abnormal genetic studies, the most frequent being Down syndrome (23/212, 10.8%), followed by DiGeorge syndrome (11/212, 5.2%). CONCLUSIONS: Prenatal screening and diagnosis of CHD in Mexico are feasible, with suspected cardiac abnormality being the main reason for referral to a specialized Fetal Cardiology Unit. Efforts must be made to make screening available to the general population in the first and second trimesters of pregnancy by fetal medicine or trained specialists, in order to identify fetal CHD and offer advanced echocardiography, genetic studies, timely fetal cardiac intervention in selected cases, and delivery in tertiary centers, to improve overall survival.
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Cardiología , Cardiopatías Congénitas , Femenino , Corazón Fetal/diagnóstico por imagen , Feto , Cardiopatías Congénitas/diagnóstico por imagen , Cardiopatías Congénitas/epidemiología , Humanos , México/epidemiología , Embarazo , Diagnóstico Prenatal , Estudios Prospectivos , Ultrasonografía PrenatalRESUMEN
BACKGROUND: Fetal aortic stenosis (AoS) may progress to hypoplastic left heart syndrome (HLHS) in utero. There are currently no data, prenatal or postnatal, describing survival of fetuses or neonates with AoS or HLHS in a country with suboptimal postnatal management. STUDY DESIGN: Prospective cohort study performed in Mexico, including cases diagnosed with AoS and HLHS within a 6-year period. AoS patients fulfilling previously published criteria for evolving HLHS (eHLHS) were offered fetal aortic valvuloplasty. Outcome variables were perinatal mortality, postnatal management, type of postnatal circulation, and overall survival. RESULTS: Fifty-four patients were included: 16 AoS and 38 HLHS. Eighteen patients had associated anomalies and/or an abnormal karyotype. Seventy-four percent of HLHS received comfort measures, with only three cases reporting an attempt at surgical palliation, and one survivor of the first stage. Fetal aortic valvuloplasty was performed successfully in nine cases of eHLHS. Overall postnatal survival was 44% in AoS with fetal aortic valvuloplasty, and one case (ongoing) in the HLHS group. CONCLUSIONS: HLHS in Mexico carries more than a 95% risk of postnatal death, with little or no experience at surgical palliation in most centers. Fetal aortic valvuloplasty in AoS may prevent progression to HLHS and in this small cohort was associated with ≈50% survival.
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Valvuloplastia con Balón , Corazón Fetal/cirugía , Fetoscopía , Síndrome del Corazón Izquierdo Hipoplásico/cirugía , Atención Posnatal/estadística & datos numéricos , Resultado del Embarazo/epidemiología , Adulto , Estenosis de la Válvula Aórtica/epidemiología , Estenosis de la Válvula Aórtica/patología , Estenosis de la Válvula Aórtica/cirugía , Valvuloplastia con Balón/métodos , Valvuloplastia con Balón/estadística & datos numéricos , Estudios de Cohortes , Circulación Coronaria/fisiología , Femenino , Corazón Fetal/patología , Fetoscopía/efectos adversos , Fetoscopía/rehabilitación , Fetoscopía/estadística & datos numéricos , Edad Gestacional , Humanos , Síndrome del Corazón Izquierdo Hipoplásico/diagnóstico , Síndrome del Corazón Izquierdo Hipoplásico/epidemiología , Recién Nacido , Masculino , México/epidemiología , Mortalidad Perinatal , Atención Posnatal/normas , Complicaciones Posoperatorias/epidemiología , Complicaciones Posoperatorias/etiología , Embarazo , Atención Prenatal/métodos , Atención Prenatal/estadística & datos numéricos , Estudios Retrospectivos , Análisis de Supervivencia , Adulto JovenRESUMEN
Risk of alcoholic cirrhosis is determined by genetic and environmental factors. We aimed to investigate if climate has a causal effect on alcohol consumption and its weight on alcoholic cirrhosis. We collected extensive data from 193 sovereign countries as well as 50 states and 3,144 counties in the United States. Data sources included World Health Organization, World Meteorological Organization, and the Institute on Health Metrics and Evaluation. Climate parameters comprised Koppen-Geiger classification, average annual sunshine hours, and average annual temperature. Alcohol consumption data, pattern of drinking, health indicators, and alcohol-attributable fraction (AAF) of cirrhosis were obtained. The global cohort revealed an inverse correlation between mean average temperature and average annual sunshine hours with liters of annual alcohol consumption per capita (Spearman's rho -0.5 and -0.57, respectively). Moreover, the percentage of heavy episodic drinking and total drinkers among population inversely correlated with temperature -0.45 and -0.49 (P < 0.001) and sunshine hours -0.39 and -0.57 (P < 0.001). Importantly, AAF was inversely correlated with temperature -0.45 (P < 0.001) and sunshine hours -0.6 (P < 0.001). At a global level, all included parameters in the univariable and multivariable analysis showed an association with liters of alcohol consumption and drinkers among population once adjusted by potential confounders. In the multivariate analysis, liters of alcohol consumption associated with AAF. In the United States, colder climates showed a positive correlation with the age-standardized prevalence of heavy and binge drinkers. Conclusion: These results suggest that colder climates may play a causal role on AAF mediated by alcohol consumption.
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Consumo de Bebidas Alcohólicas/efectos adversos , Clima Frío/efectos adversos , Cirrosis Hepática Alcohólica/epidemiología , Consumo de Bebidas Alcohólicas/epidemiología , Estudios Transversales , Humanos , Internacionalidad , Cirrosis Hepática Alcohólica/etiología , Luz Solar , Estados Unidos/epidemiologíaRESUMEN
OBJECTIVES: The objectives of the study are to evaluate longitudinal changes in lung size and intrapulmonary-artery (IPa) Doppler in fetuses with congenital diaphragmatic hernia (CDH) and assess their contribution in predicting neonatal survival. METHODS: The observed/expected lung-to-head ratio (O/E-LHR) and IPa-pulsatility index (PI) and peak early diastolic reversed flow (PEDRF) were evaluated in a cohort of left-sided CDH fetuses managed expectantly during pregnancy. Longitudinal changes were analyzed by multilevel analysis, and their value to predict survival using the multiple logistic regression and decision-tree analysis was assessed. RESULTS: A total of 232 scans were performed on 69 CDH fetuses. The O/E-LHR values remained unchanged during fetal monitoring, whereas IPa-PI and PEDRF showed a progressive increase throughout follow-up, becoming abnormal on average at 30 weeks of gestation. Absent/reversed end-diastolic velocity (EDV) in the IPa was observed in 20.3%. O/E-LHR and IPa Doppler indices were significantly associated with probability of survival (O/E-LHR ≥ 26%, odds ratio [OR] 19.0; IPa-PI <+2.0 z score, OR 3.0; and positive EDV, OR 7.4). All cases with IPa-reversed EDV died after birth. CONCLUSION: While lung size remains stable during pregnancy, CDH fetuses show progressive deterioration in intrapulmonary blood flow. IPa Doppler evaluation may aid in predicting survival of CDH fetuses managed expectantly during pregnancy.
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Edad Gestacional , Hernias Diafragmáticas Congénitas/embriología , Pulmón/diagnóstico por imagen , Pulmón/embriología , Arteria Pulmonar/diagnóstico por imagen , Ultrasonografía Prenatal , Femenino , Hernias Diafragmáticas Congénitas/diagnóstico por imagen , Hernias Diafragmáticas Congénitas/mortalidad , Humanos , Recién Nacido , Pulmón/irrigación sanguínea , Embarazo , Tasa de Supervivencia , Ultrasonografía DopplerRESUMEN
OBJECTIVE: To evaluate cardiac function by conventional echocardiography and tissue Doppler imaging in fetuses with left congenital diaphragmatic hernia (CDH). METHODS: Conventional echocardiography (myocardial performance index, ventricular filling velocities, and E/A ratios) and tissue Doppler imaging (annular myocardial peak velocities, E/E' and E'/A' ratios) in mitral, septal, and tricuspid annulus were evaluated in a cohort of 31 left-sided CDH fetuses and compared with 75 controls matched for gestational age 2:1. RESULTS: In comparison to controls, CDH fetuses had prolonged isovolumetric time periods (isovolumetric contraction time 35 ms vs 28 ms, P < .001), with higher myocardial performance index (0.49 vs 0.42, P < .001) and tricuspid E/A ratios (0.77 vs 0.72, P = .033). Longitudinal function assessed by tissue Doppler showed signs of impaired relaxation (mitral lateral A' 8.0 vs 10.1 cm/s, P < .001 and an increased mitral lateral E'/A' ratio 0.93 vs 0.78, P < .001) in the CDH fetuses as compared with controls, with preserved systolic function. CONCLUSION: Left CDH fetuses show echocardiographic signs of diastolic dysfunction, probably secondary to fetal heart compression, maintaining a preserved systolic function.
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Corazón Fetal/fisiopatología , Hernias Diafragmáticas Congénitas/fisiopatología , Adulto , Estudios de Casos y Controles , Ecocardiografía , Femenino , Corazón Fetal/diagnóstico por imagen , Hernias Diafragmáticas Congénitas/diagnóstico por imagen , Humanos , Embarazo , Estudios Prospectivos , Ultrasonografía PrenatalRESUMEN
INTRODUCTION: Fetal growth restriction (FGR) affects 5%-10% of all pregnancies, contributing to 30%-50% of stillbirths. Unfortunately, growth restriction often is not detected antenatally. The last weeks of pregnancy are critical for preventing stillbirth among babies with FGR because there is a pronounced increase in stillbirths among growth-restricted fetuses after 37 weeks of pregnancy. Here we present a protocol (V.1, 23 May 2016) for the RATIO37 trial, which evaluates an integrated strategy for accurately selecting at-risk fetuses for delivery at term. The protocol is based on the combination of fetal biometry and cerebroplacental ratio (CPR). The primary objective is to reduce stillbirth rates. The secondary aims are to detect low birth weights and adverse perinatal outcomes. METHODS AND ANALYSIS: The study is designed as multicentre (Spain, Chile, Mexico,Czech Republic and Israel), open-label, randomised trial with parallel groups. Singleton pregnancies will be invited to participate after routine second-trimester ultrasound scan (19+0-22+6 weeks of gestation), and participants will be randomly allocated to receive revealed or concealed CPR evaluation. Then, a routine ultrasound and Doppler scan will be performed at 36+0-37+6 weeks. Sociodemographic and clinical data will be collected at enrolment. Ultrasound and Doppler variables will be recorded at 36+0-37+6 weeks of pregnancy. Perinatal outcomes will be recorded after delivery. Univariate (with estimated effect size and its 95% CI) and multivariate (mixed-effects logistic regression) comparisons between groups will be performed. ETHICS AND DISSEMINATION: The study will be conducted in accordance with the principles of Good Clinical Practice. This study was accepted by the Clinical Research Ethics Committee of Hospital Clinic Barcelona on 23May 2016. Subsequent approval by individual ethical committees and competent authorities was granted. The study results will be published in peer-reviewed journals and disseminated at international conferences. TRIAL REGISTRATION NUMBER: NCT02907242; pre-results.
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Retardo del Crecimiento Fetal/diagnóstico por imagen , Monitoreo Fetal , Insuficiencia Placentaria/diagnóstico por imagen , Complicaciones del Embarazo/diagnóstico por imagen , Tercer Trimestre del Embarazo , Mortinato/epidemiología , Adulto , Chile , República Checa , Femenino , Retardo del Crecimiento Fetal/mortalidad , Retardo del Crecimiento Fetal/fisiopatología , Humanos , Recién Nacido , Israel , México , Insuficiencia Placentaria/mortalidad , Insuficiencia Placentaria/fisiopatología , Embarazo , Complicaciones del Embarazo/fisiopatología , Resultado del Embarazo , Atención Prenatal/estadística & datos numéricos , España , Ultrasonografía Prenatal/estadística & datos numéricos , Adulto JovenRESUMEN
Mobilized peripheral blood (MPB) bone marrow cells possess the potential to differentiate into a variety of mesenchymal tissue types and offer a source of easy access for obtaining stem cells for the development of experimental models with applications in tissue engineering. In the present work, we aimed to isolate by magnetic activated cell sorting CD90+ cells from MPB by means of the administration of Granulocyte-Colony Stimulating Factor and to evaluate cell proliferation capacity, after thawing of the in vitro culture of this population of mesenchymal stem cells (MSCs) in sheep. We obtained a median of 8.2 ± 0.6 million of CD90+ cells from the 20-mL MPB sample. After thawing, at day 15 under in vitro culture, the mean CD90+ cells determined by flow cytometry was 92.92 ± 1.29 % and cell duplication time determined by crystal violet staining was 47.59 h. This study describes for the first time the isolation, characterization, and post-in vitro culture thawing of CD90+ MSCs from mobilized peripheral blood in sheep. This population can be considered as a source of MSCs for experimental models in tissue engineering research.
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Criopreservación/métodos , Células Madre Mesenquimatosas/citología , Células Madre de Sangre Periférica/citología , Antígenos Thy-1/metabolismo , Animales , Diferenciación Celular , Proliferación Celular , Separación Celular , Forma de la Célula , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Inmunofenotipificación , Masculino , OvinosRESUMEN
OBJECTIVE: To evaluate Doppler parameters of the two segments of the posterior cerebral artery (PCA) in normally grown and growth restricted (IUGR) fetuses. METHODS: Normal reference values of the pulsatility index (PI), peak systolic velocity, end-diastolic velocity, and time-averaged maximum velocity from the two segments of the PCA (segment one, PCA-S1; segment two, PCA-S2) were constructed in 350 pregnant women from 20 to 40 weeks of gestation. The association of the two PCA segments with the middle cerebral artery (MCA), and with the umbilical artery was evaluated in 50 IUGR fetuses. RESULTS: In normal fetuses, Doppler parameters from PCA-S1 and PCA-S2 showed similar behavior throughout gestation. In IUGR fetuses, the PI of the two PCA segments was significantly reduced with no differences between them. PCA-S2 PI showed a higher correlation with the MCA-PI (r = 0.73) than PCA-S1 PI (r = 0.63; p = 0.001). IUGR fetuses showed vasodilatation in the PCA earlier than in the MCA disregarding the umbilical artery-PI value. CONCLUSION: In IUGR fetuses, the two segments of the PCA show signs of vasodilatation earlier than the MCA. As IUGR fetuses deteriorate, the two segments of the PCA and the MCA behave similarly.
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Retardo del Crecimiento Fetal/diagnóstico por imagen , Arteria Cerebral Media/diagnóstico por imagen , Arteria Cerebral Posterior/diagnóstico por imagen , Arterias Umbilicales/diagnóstico por imagen , Adolescente , Adulto , Velocidad del Flujo Sanguíneo , Estudios de Casos y Controles , Femenino , Humanos , Embarazo , Flujo Pulsátil , Ultrasonografía Doppler , Ultrasonografía Prenatal , Adulto JovenRESUMEN
OBJECTIVE: To assess the value of gestational age and cardiovascular Doppler indices in predicting perinatal mortality in a multicenter cohort of early-onset intrauterine growth-restricted (IUGR) fetuses. METHODS: A multicenter prospective cohort study including 157 early-onset (<34 weeks) IUGR cases with abnormal umbilical artery (UA) Doppler was conducted. Cardiovascular assessment included the ductus venosus (DV), the aortic isthmus flow index (IFI), and the myocardial performance index (MPI). Isolated and combined values to predict the risk of perinatal death were evaluated by logistic regression and by decision tree analysis, where the gestational age at delivery, UA, and middle cerebral artery (MCA) were also included as covariates. RESULTS: Perinatal mortality was 17% (27/157). All parameters were significantly associated with perinatal death, with individual odds ratios (OR) of 25.2 for gestational age below 28 weeks, 12.1 for absent/reversed DV atrial flow, 5.3 for MCA pulsatility index <5th centile, 4.6 for UA absent/reversed diastolic end-flow, 1.8 for IFI <5th centile, and 1.6 for MPI >95th centile. Decision tree analysis identified gestational age at birth as the best predictor of death (<26 weeks, 93% mortality; 26-28 weeks, 29% mortality, and >28 weeks, 3% mortality). Between 26 and 28 weeks, DV atrial flow allowed further stratification between high (60%) and low risk (18%) of mortality. CONCLUSIONS: Gestational age largely determines the risk of perinatal mortality in early-onset IUGR before 26 weeks and later than 28 weeks of gestation. The DV may improve clinical management by stratifying the probability of death between 26 and 28 weeks of gestation.