Generation of a novel three-dimensional scaffold-based model of the bovine endometrium

Bovine in vitro endometrial models that resemble tissue function in vivo are needed to study infertility, long-term uterine alterations induced by pathogens and impact of endocrine disruptor chemicals on reproductive function and other reproductive system complications that cause high economic losses in livestock species. The present study aimed to generate an innovative, reproducible, and functional 3D scaffold-based model of the bovine endometrium structurally robust for long term-culture. We developed a multicellular model containing both endometrial epithelial and stromal cells. Epithelial cells organized to form a luminal-like epithelial layer on the surface of the scaffold. Stromal cells produced their own extracellular matrix forming a stable subepithelial compartment that physiologically resembles the normal endometrium. Both cell types released prostaglandin E2 and prostaglandin F2α following a treatment with oxytocin and arachidonic acid. Additionally signal pathways mediating oxytocin and arachidonic acid stimulation of prostaglandin synthesis were analyzed by real time PCR (RT-PCR). Oxytocin receptor (OXTR), prostaglandin E2 receptor 2 (EP2), prostaglandin E2 receptor 4 (EP4), prostaglandin F receptor (PTGFR), prostaglandin E synthase (PTGES), PGF-synthase (PGFS) and prostaglandin-endoperoxide synthase 2 (COX-2) expression was detected in both control and treatment groups, however, only significant changes in abundance of OXTR mRNA transcripts were found. The results obtained by this study are a step forward in bovine in vitro culture technology. This 3D scaffold-based model provides a platform to study regulatory mechanisms involved in endometrial physiology and can set the basis for a broader tool for designing and testing novel therapeutic strategies for recurrent uterine pathologies

Citrato de cafeína: ¿por qué usarlo en los recién nacidos?

ResumenEl citrato de cafeína, es uno de los medicamentos más comúnmente usado en cualquier unidad de cuidados intensivos neonatales; es una metilxantina e inhibidor de los receptores de adenosina. Desde hace más de 30 años es bien conocida su eficacia en el tratamiento de la apnea del prematuro, actualmente se le reconocen otros beneficios como disminución en la incidencia de displasia broncopulmonar, disminución de la falla a la extubación y ser un factor de neuroprotección entre otros. El objetivo del presente artículo fue realizará una revisión sobre las propiedades farmacológicas, mecanismos de acción, dosis adecuadas, niveles séricos y efectos terapéuticos de la cafeína en los recién nacidos.AbstractCaffeine, a methylxantine and nonspecific inhibitor of adenosine receptors, is commonly prescribed in the Neonatal Intensive Care Units. The efficacy of caffeine in an episode of apnea of prematurity has been known for over thirty years, but now it has been found that caffeine have others beneficial actions, such as reduction in the incidence of bronchopulmonary dysplasia, less extubation failure, the need for patent ductus arteriosus treatment, and neuroprotective role. The objective is this review was to update the current knowledge of pharmacologic properties, the mechanisms of action, caffeine dosing and therapeutic effects, in the neonatal population

Comparative study of PCR-sexing procedures using bovine embryos fertilized with sex-sorted spermatozoa

Sex determination in bovine embryos is a useful tool in reproductive biotechnology. This work compares two techniques of embryo sexing by polymerase chain reaction (PCR. Embryos were produced in vitro with sex-sorted sperm and two techniques of DNA lysis were tested (proteinase K versus heat shock). Subsequently, halves of each lysed sample was amplified both by amelogenin and BRY4a/SAT1 primers. Male embryos treated by both digestion methods and amplified by BRY4a/SAT1 gave higher rates of false negatives. Amelogenin amplification failed with embryos previously digested by proteinase K. In contrast, the lysis method allowed obtaining the best accuracy in terms of sex verification when using amelogenin amplification

Gene expression in early expanded parthenogenetic and in vitro fertilized bovine blastocysts.

Mammalian oocytes can undergo artificial parthenogenesis in vitro and develop to the blastocyst stage. In this study, using real-time PCR, we analyzed the expression of genes representative of essential events in development. In vitro matured oocytes were either fertilized or activated with ionomycin + 6-DMAP and cultured in simple medium. The pluripotency-related gene Oct3/4 was downregulated in parthenotes, while the de novo methylation DNMT3A gene was unchanged. Among the pregnancy recognition genes, IFN-t was upregulated, PGRMC1 was downregulated and PLAC8 was unchanged in parthenotes. Among the metabolism genes, SLC2A1 was downregulated, while AKR1B1, COX2, H6PD and TXN were upregulated in parthenotes; there was no difference in SLC2A5. Among the genes involved in compaction/blastulation, GJA1 expression increased in parthenotes, but no differences were detected within ATP1A1 and CDH1. Expression of p66(shc) and the Bax/Bcl2 ratio were higher in parthenotes, and there was no difference in p53. Parthenotes and embryos may differ in the way they stimulate apoptosis, with a preponderant role for p66(shc) within parthenotes. Differentially affected functions may also include pluripotency, de novo methylation and early embryonic signalling

Valores de saturación periférica de oxígeno por oximetría de pulso en recién nacidos de término sin patología respiratoria

Resumen: Introducción: La saturación es un determinante fisiológico básico que refleja la entrega de oxígeno tisular, y se ha convertido en una herramienta indispensable en neonatología, en particular en los pacientes que reciben oxigenoterapia. Objetivo: Obtener los valores de saturación periférica por oximetría de pulso en recién nacidos de término sin patología respiratoria de la terapia de invasión mínima (TIM) y compararlos con los recién nacidos de término ingresados a alojamiento conjunto. Material y métodos: Se realizó un estudio observacional, transversal analítico en neonatos de término, nacidos en el Instituto Nacional de Perinatología. Los valores de oximetría de pulso se tomaron desde las primeras 24 h de vida, por 2 días. Los datos fueron procesados en una base de datos del programa SPSS, versión 13. Resultados: No se encontraron diferencias estadísticamente significativas entre el comportamiento de la saturación de oxígeno durante el día entre los grupos. En el grupo de recién nacidos ingresados a la TIM se observa una disminución importante de los niveles de oxígeno por oximetría de pulso a los 5 min de iniciada la alimentación. Conclusiones: Los recién nacidos ingresados a alojamiento conjunto presentan menos desaturaciones en comparación con los recién nacidos de la TIM porque son alimentados al seno materno de manera exclusiva, lo que promueve una mejor coordinación en la succión, deglución y respiración, situación que se refleja en los niveles sanguíneos de saturación. Abstract: Introduction: Saturation is a basic physiological factor that reflects the delivery of oxygen to the tissues, and has become a crucial tool in neonatology, particularly especially in patients receiving oxygen therapy. Objective: To obtain the values of peripheral saturation by pulse oximetry in term newborns without respiratory disease and compare them with the term newborns admitted to the Mother and Baby Unit. Material and methods: An observational, cross-sectional analytical study was carried out on term neonates, born in the National Institute of Perinatology. The pulse oximetry values were taken in the first hours of life, for up to 2 days. The data were processed using the SPSS program, version 13. Results: No statistically significant differences were found between the oxygen saturation patterns during the day, or between the groups. In the group of newborns admitted to the Minimal Invasion Therapy (MIT) unit, a significant decrease in oxygen levels was observed in the pulse oximetry 5 minutes after starting feedings. Conclusions: Newborns admitted to the Mother and Baby Unit have less desaturation compared to newborns in the MIT. This may be because they are exclusively breastfed, which promotes better coordination in sucking, swallowing and breathing, a situation that is reflected in the saturation blood levels. Palabras clave: Saturación, Oxígeno, Oximetría de pulso, Recién nacidos, Keywords: Saturation, Oxygen, Pulse oximetry, Newborn

Estudio comparativo de dos métodos de sexaje por PCR en embriones bovinos producidos con semen sexado

Sex determination in bovine embryos is a useful tool in reproductive biotechnology. This work compares two techniques of embryo sexing by polymerase chain reaction (PCR). Embryos were produced in vitro with sex-sorted sperm and two techniques of DNA lysis were tested (proteinase K versus heat shock). Subsequently, halves of each lysed sample was amplified both by amelogenin and BRY4a/SAT1 primers. Male embryos treated by both digestion methods and amplified by BRY4a/SAT1 gave higher rates of false negatives. Amelogenin amplification failed with embryos previously digested by proteinase K. In contrast, the lysis method allowed obtaining the best accuracy in terms of sex verification when using amelogenin amplification.La determinación del sexo en embriones bovinos es una herramienta útil en las biotecnologías reproductivas. Este trabajo compara dos técnicas de determinación del sexo en embriones mediante la reacción en cadena de la polimerasa (PCR). Los embriones fueron producidos in vitro con semen sexado y se analizaron dos técnicas de lisis de ADN (proteinasa K y choque térmico). Posteriormente, la mitad de cada muestra lisada fue amplificada por la amelogenina y por los cebadores BRY4a/SAT1. Los embriones machos tratados por ambos métodos de digestión y amplificados por BRY4a/SAT1 dieron mayores tasas de falsos negativos. La amplificación de la amelogenina no dio resultados en embriones previamente digeridos por proteinasa K; sin embargo, el método de lisis por choque térmico junto con la amplificación por amelogenina permitió obtener la mayor precisión en términos de verificación de sexo