Prolactin Protects the Structural Integrity of Human Fetal Membranes by Downregulating Inflammation-induced Secretion of Matrix Metalloproteinases.

Prolactin (PRL) is a pleiotropic hormone with a key role in pregnancy. In fetal membranes, PRL can regulate the secretion of pro-inflammatory factors, which induces the activation of matrix metalloproteinases (MMPs). The increase and activation of MMPs deregulate the turnover of the extracellular matrix in the fetal membranes, altering its structure and function, causing premature rupture of the membranes and preterm labor. In this work, we evaluate the effect of PRL upon the secretion of MMP-1, MMP-2, MMP-9, MMP-13, and the tissue inhibitors of metalloproteinases (TIMPs) in human fetal membranes after lipopolysaccharide (LPS) challenge. Nine fetal membranes from healthy non-laboring cesarean deliveries at term were cultured in a 2-independent chamber system and pre-treated with 250, 500, 1000 or 4000 ng/ml of PRL for 24 h, then choriodecidual region was stimulated with 500 ng/ml of LPS plus fresh PRL for 24 h. The MMPs and TIMPs secretion were quantified by ELISA, additionally MMP-2 and MMP-9 gelatinolytic activity was measured by zymography. LPS induced the MMP-9 and MMP-1 secretion, but no MMP-2 or MMP-13 in comparison with basal levels. PRL co-treatment decreased the MMP-2, MMP-9 and MMP-1 secretion induced by LPS. The active forms were present in the tissue extract, showing a response consistent with the secretion profile. TIMP-1 and TIMP-2 secretion was decreased after LPS treatment and the PRL co-treatment reverts this effect. The present results support that PRL may favor the balance between these factors involved in the structural maintenance of fetal membranes in an inflammatory event.

[Molecular identification of glucose-6-phosphate dehydrogenase (G6PD) detected in neonatal screening]. / Identificación molecular de la glucosa-6-fosfato deshidrogenasa (G6PD) detectada en el tamiz neonatal.

AIMS: To present the strategy of identifying the molecular variants of G6PD detected in neonatal screening (NS). MATERIAL AND METHODS: We present a series of incident cases of newborns positive for G6PD deficiency detected in NS. From nuclear DNA with the methodology of real-time PCR we sought molecular G6PD variants: G202A, A376G, T968C and C563T. RESULTS: Of a total of 21,619 neonates, 41 cases were reactive in NS for G6PD (189.6/100,000 RN screened rate), 34 cases confirmed the molecular variant of G6PD (157.3/100,000 RN screened rate). The most frequent allele combination G202A/A376G (G6PD ratio and median activity, 0.460 and 1.72 ± 0.35 U/g Hb, respectively), followed by G202A (0.170 and 1.74 ± 0.27 U/g Hb) and A376G/T968C (ratio 0.150 and 1.10 ± 0.44 U/g Hb). The T968C allelic variant showed lower enzyme activity than the rest (1.1 ± 0.4; p = 0.02). Two women were detected with G6PD deficiency with G202A/A376G and G202A variant. CONCLUSIONS: African alleles were prevalently detected in neonatal screening. This strategy allows the identification of molecular variants involved in 80% of cases.

Effect of Oropharyngeal Administration of Colostrum in Premature Newborns ≤32 Weeks of Gestation on the Immune Response and Neonatal Morbidity: A Double-Blind Randomized Clinical Trial.

Introduction: The mother's colostrum carries immunological components, such as cytokines and immunoglobulins (Igs), derived from the maternal circulation with bacteriostatic properties. Objective: The objective of this study was to evaluate the effect of oropharyngeal administration of colostrum (OPAC) vs. placebo in the first 4 days of life in premature newborns ≤32 weeks of gestation on serum Ig concentration, neonatal morbidity, and total days of hospitalization. Hypothesis: The OPAC increases serum Igs and decreases morbidity and total days of hospitalization. Materials and Methods: A double-blind randomized controlled trial was carried out. Participants were randomly assigned to one of the two groups, namely, group 1: placebo (P) (n = 50) and group 2: colostrum (C) (n = 46). A blood sample was obtained at baseline and 7 and 28 days of life to quantify immunoglobulin G (IgG), immunoglobulin A (IgA), and IgM. Results: The C group showed an increase in serum IgA on day 28 expressed as median and [interquartile range]; C: 25 [12-35] vs. P: 11 [8-18], p < 0.001. There were no significant differences in neonatal morbidity. Newborns in the colostrum group showed the completed enteral feeding earlier (days), C: 13.9 ± 7 vs. P: 17.4 ± 8.4, p < 0.04; they reached the birth weight earlier, C: 10.9 ± 2.8 vs. P: 12.9 ± 4, p < 0.01, and had less days of hospitalization, C: 60.2 ± 33.8 vs. P: 77.2 ± 47.3, p < 0.04. Neonatal mortality was lower in the colostrum group than the placebo group 0% vs. 12%, respectively, without a statistical difference (p = 0.06). Conclusion: In premature newborns ≤32 weeks of gestation, the OPAC within 4 days after birth increases serum IgA concentration at day 28 compared to placebo. Similarly, OPAC decreased the days to complete enteral feeding and reach the birth weight and total days of hospitalization. Clinical Trial Registration: [https://clinicaltrials.gov/ct2/show/NCT03578341], identifier: [NCT03578341].

Prolactin modifies the in vitro LPS-induced chemotactic capabilities in human fetal membranes at the term of gestation.

PROBLEM: Immune responses of fetal membranes involve the production of chemoattractant mediators causing infiltration of maternal and fetal leukocytes, intrauterine inflammation and potentially the disruption of maternal-fetal tolerance. Prolactin (PRL) has deep immunoregulatory effects in the fetal-maternal interface. We aimed to test the in vitro PRL effect upon chemotactic capacities of human fetal membranes. METHOD OF STUDY: Fetal membranes and umbilical cord blood were collected from healthy non-laboring caesarean deliveries at term. Fetal membranes were cultured in Transwell® frames to mimic the barrier function between choriodecidual and amniotic sides. Tissues were treated with PRL, Lipopolysaccharide (LPS), or both simultaneously. Then, RANTES, MCP-1, MIP-1α, IP-10, and PECAM-1 were quantified in a conditioned medium by choriodecidual or amniotic sides. The chemotaxis of subsets of migrating mononuclear cells from umbilical cord blood was evaluated in a Boyden Chamber in response to the conditioned medium by both sides. RESULTS: Lipopolysaccharide stimulates the production of RANTES, MCP-1, MIP-1α, and PECAM-1 in choriodecidua, while MIP-1α and PECAM-1 only increase in amnion. PRL decrease RANTES, MCP-1, and MIP-1 only in choriodecidua, but PECAM-1 was decreased mainly in amnion. The leukocyte migration was regulated significantly in response to the conditioned medium by the amnion, increase in the conditioned medium after LPS treatment, contrary with, the leukocyte migration decreased in a significant manner in response to conditioned medium after PRL and LPS-PRL co-treatment. Finally, T cells were the most responsive subset of cells. CONCLUSIONS: Prolactin modified in a tissue-specific manner the chemotactic factor and the leukocyte migration differentially in fetal membranes.

Multiparameter flow cytometry analysis of leukocyte markers for diagnosis in preterm neonatal sepsis.

PURPOSE: Neonatal sepsis is an important public health concern worldwide due to its immediate lethality and long-term morbidity rates, Clinical evaluation and laboratory analyses are indispensable for diagnosis of neonatal sepsis. However, assessing multiple biomarkers in neonates is difficult due to limited blood availability. The aim is to investigate if the neonatal sepsis in preterm could be identified by multiparameter analysis with flow cytometry. MATERIALS AND METHODS: The expression of activation-related molecules was evaluated by flow cytometry in newborn with or without risk factors for sepsis. RESULTS: Our analysis revealed that several markers could be useful for sepsis diagnosis, such as CD45RA, CD45RO, or CD71 on T cells; HLA-DR on NKT or classic monocytes, and TREM-1 on non-classic monocytes or neutrophils. However, ROC analysis shows that the expression of CD45RO on T lymphocytes is the only useful biomarker for diagnosis of neonatal late-onset sepsis. Also, decision tree analyses showed that CD45RO plus CD27 could help differentiate the preterm septic neonates from those with risk factors. CONCLUSIONS: Our study shows a complementary and practical strategy for biomarker assessment in neonatal sepsis.

Neurodesarrollo en infantes con antecedente de hipotiroidismo congénito / Neurodevelopment in infants with congenital hypothyroidism antecedents

Introducción: Los estudios de seguimiento de infantes con hipotiroidismo congénito con tratamiento temprano, muestran que no existen diferencias con respecto al coeficiente intelectual que se encuentra en la población general. Material y métodos: Estudio de cohorte retrospectivo. Veinticuatro hipotiroideos fueron detectados a través del Programa de Tamiz Neonatal y comparados con un grupo testigo. Los recién nacidos fueron valorados entre los seis meses y los siete años de edad mediante el índice de desarrollo (ID) de Bayley, el coeficiente intelectuales (Cl) de Terman Merril y la prueba Hiskey Nebraska para detectar infantes con hipoacusia. Resultados: Dieciocho infantes hipotiroideos mostraron Cl o ID normal o alto. Uno con ID de Bayley porabajo del promedio, dos con Cl por abajo del promedio y uno con hipoacusia y capacidad de aprendizaje baja. No se encontraron diferencias con respecto al grupo control. El uso de ototóxicos, prematurez e hiperbilirrubinemia, fueron las causas que explicaron los coeficientes bajos entre los hipotiroideos y el grupo control. Conclusiones: El Cl y el ID en el grupo de hipotiroidismo tratado tempranamente no difirió con los resultados del grupo testigo.

Introduction: Follow-up studies of hypothyroid infants who received early treatment evidence the presence of the same IQ vs. the general population. Material and methods: Retrospective cohort study. Twenty-four hypothyroid infants detected in neonatal screening were compared to a control group. The newborns were assessed from six months to seven years based on Bayley's infant development scales (ID), the Terman-Merril IQ (Cl) and the learning aptitude of the heard of hearing with the Hiskey-Nebraska test (CA). Results: Twenty hypothyroid infants showed a normal or high Cl or ID. One of the infants with a ID result below the mean, two with an Cl below the mean, one hearing impaired infant with low learning aptitude. No differences were found vs. the control group. The use of ototoxic medications, prematurity and hyperbilirubinemia explain the low coefficients between the hypothyroid individuals and the control. Conclusions: The Cl and the ID results in the hypothyroid group who received early treatment showed no difference vs. the control group.

Neurodesarrollo a los 3 años de vida en neonatos sometidos a ventilación mecánica / Neurodevelopment at the age of three in neonates subjected to mechanical ventilation

Introducción. Una de las principales preocupaciones de las Unidades de Cuidades Intensivos Neonatales es la calidad de vida de sus egresados, en varios estudios se reporta un aumento significativo en la supervivencia de los mismos en los últimos 12 años. La ventilación mecánica convencional, procedimiento habitual en estos neonatos, se ha asociado en varios trabajos a alteraciones del desarrollo en general. Material y métodos. Se estudiaron en forma retrospectiva a un grupo de neonatos nacidos en el período de enero de 1991 a enero de 1993 sometidos a ventilación mandatoria intermitente desde su nacimiento hasta un mínimo de 3 días, encontrándose todos dentro del Programa de seguimiento Pediátrico y con valoraciones: neurológica, psicológica, motora, auditiva, de lenguaje y oftalmológica. Resultados. Ciento noventa y siete pacientes reunieron los criterios de inclusión. El resultado global de la valoración neurológica mostró un predominio de niños normales del 59 por ciento, con alteraciones moderadas del 16 por ciento y graves del 9 por ciento. Las alteraciones psicológicas fueron del 33 por ciento, de acuerdo al coeficiente intelectual (IQ) el 66 por ciento se encontró igual o mayor al promedio. La audición mostró 59 por ciento de normalidad, con alteraciones entre graves y profundas del 19 por ciento. La valoración del lenguaje presentó un porcentaje alto de afectados del 51 por ciento; sin embargo, sólo 22 por ciento tenía alteraciones graves en relación directa con la pérdida auditiva mencionada. En la valoración oftalmológica se encontró una proporción de 0.81 de pacientes sanos. Conclusiones. La interpretación a la asociación de la ventilalción mecánica, variables neonatales y desarrollo posterior sólo pueden ser parciales ya que el conocimiento y el manejo de todas las variables implícitas en este complejo proceso es sumamente difícil