Protocol for a multicenter, double-blind, randomized, placebo-controlled phase III trial of the inhaled ß2-adrenergic receptor agonist salbutamol for transient tachypnea of the newborn (the REFSAL trial).

Background: Transient tachypnea of the newborn (TTN), which results from inadequate absorption of fetal lung fluid, is the most common cause of neonatal respiratory distress. Stimulation of ß-adrenergic receptors enhances alveolar fluid absorption. Therefore, the ß2-adrenergic receptor agonist salbutamol has been proposed as a treatment for TTN. This study aims to evaluate the efficacy and safety of salbutamol as supportive pharmacotherapy together with non-invasive nasal continuous positive airway pressure (NIV/nCPAP) for the prevention of persistent pulmonary hypertension of the newborn (PPHN) in infants with TTN. Methods and analysis: This multicenter, double-blind, phase III trial will include infants with a gestational age between 32 and 42 weeks who are affected by respiratory disorders and treated in eight neonatal intensive care units in Poland. A total of 608 infants within 24 h after birth will be enrolled and randomly assigned (1:1) to receive nebulized salbutamol with NIV or placebo (nebulized 0.9% NaCl) with NIV. The primary outcome is the percentage of infants with TTN who develop PPHN. The secondary outcomes are the severity of respiratory distress (assessed with the modified TTN Silverman score), frequency of need for intubation, duration of NIV and hospitalization, acid-base balance (blood pH, partial pressure of O2 and CO2, and base excess), and blood serum ionogram for Na+, K+, and Ca2+. Discussion: The Respiratory Failure with Salbutamol (REFSAL) study will be the first clinical trial to evaluate the efficacy and safety of salbutamol in the prevention of persistent pulmonary hypertension in newborns with tachypnea, and will improve short term outcomes. If successful, the study will demonstrate the feasibility of early intervention with NIV/nCPAP together with nebulized salbutamol in the management of TTN. Ethics and dissemination: The study protocol was approved by the Bioethics Committee of the Medical University of Warsaw, Warsaw, Poland on November 16, 2020 (decision number KB/190/2020). All procedures will follow the principles of the Declaration of Helsinki. The results of the study will be submitted for knowledge translation in peer-reviewed journals and presented at national and international pediatric society conferences. Clinical Trial Registration: It is registered at ClinicalTrials.gov NCT05527704, EudraCT 2020-003913-36; Protocol version 5.0 from 04/01/2022.

The concentration of insulin-like growth factor-1 in pregnancies complicated by pregnancy-induced hypertension and/or intrauterine hypotrophy.

OBJECTIVES: The aim of the study was to compare Insulin-like Growth Factor-1 (IGF-1)concentration in pregnancies complicated by pregnancy-induced hypertension and/or intrauterine hypotrophy, and its correlation with maternal pressure and umbilical artery pulsatility and resistance indices. MATERIAL AND METHODS: 65 pairs pregnant-newborn were included to four groups: I - control, II - PIH, III - Hypotrophy, IV - PIH and Hypotrophy. In the study we analyzed cord blood IGF-1 concentration, newborns antropometry, umbilical artery pulsatility and resistance indices and maternal pressure before delivery. RESULTS: The concentration of IGF-1 was the lowest in IV group of hypotrophic newborns from pregnancies complicated by pregnancy-induced hypertension. In this group of patients there was strong negative correlation between IGF-1 concentration and maternal systolic and diastolic pressure. CONCLUSIONS: There is a strong negative correlation between IGF-1 concentration and maternal systolic pressure in group of hypotrophic newborns from pregnancies complicated by pregnancy-induced hypertension.

Influence of infections on the quality of general movements in premature infants.

INTRODUCTION: The task of contemporary neonatology is not only to save the lives of children born prematurely, but also to provide them with the highest possible quality of life. The Prechtl method, one of several methods of assessing the quality of general movement patterns, enables early identification of immaturity or damage to the central nervous system. AIM: Dynamic assessment of the type and quality of general movement (GMs) in preterm infants to determine indications for early neurodevelopmental support and to identify the relationship between the occurrence of maternal and neonatal perinatal risk factors and the type and quality of general movements. MATERIAL AND METHOD: 90 infants were assessed between 28 0/7 and 36 6/7 weeks gestational age. Dominant cases (57 cases) were infants born between 32 and 36 weeks GE. Most cases (48) received low Apgar score at 1 min (≤7), including two born in very severe condition. Neonatal infections were diagnosed in 26 infants. The study included three GM assessments: I - up to 14th day post-partum, II at term and III between 12 and 15 weeks corrected age. The analysis included gestational age, general condition of the newborn as per Apgar score as well as early and late infections. RESULTS: Neonatal infections were found to show a close correlation with the occurrence of abnormal general movements. It was confirmed that gestational age and congenital infections are important variables affecting the occurrence of abnormal general movements.

Effects of oral probiotic supplementation on gut Lactobacillus and Bifidobacterium populations and the clinical status of low-birth-weight preterm neonates: a multicenter randomized, double-blind, placebo-controlled trial.

AIM: Probiotic bacteria administered directly after birth to preterm neonates may improve gastrointestinal function and may reduce the incidence of late-onset sepsis, which is a frequent complication in this group. PURPOSE: The main objective of this study was to evaluate whether a new probiotic bacterial mixture of Lactobacillus rhamnosus KL53A and Bifidobacterium breve PB04 given to preterm, low-birth-weight neonates would influence composition of their gut microbiota and sepsis rates. PATIENTS AND METHODS: This study was a multicenter, randomized, double-blind, placebo-controlled trial conducted in clinical centers of neonatal care in Poland. A probiotic or placebo preparation was given twice daily to 181 preterm low-birth-weight neonates who were eligible for enteral feeding between July 2012 and July 2013. The probiotic was given to 90 neonates, while placebo was given to 91 neonates. The gut microbiota was monitored by microbiological analysis of stool samples. Sepsis episodes were detected on the basis of clinical and laboratory findings and confirmed by blood cultures. RESULTS: Tested probiotic administration resulted in continuous increase of the Lactobacillus and Bifidobacterium counts in the gut microbiota. The applied tested strains successfully colonized the neonates gut since they were present in over 90% of stool samples, which was confirmed by molecular analysis. Regardless of the study group (probiotic or placebo), B. breve colonization correlated with lower staphylococcal sepsis incidence, which was irrespective of whether probiotics were given. No sepsis case caused by strains included in study probiotic was recorded. CONCLUSION: Appropriately selected and characterized probiotic bacteria may be safely given to preterm neonates to normalize their distorted gut microbiota and may contribute to lower staphylococcal sepsis rates.

Monitoring the Transcriptional Activity of Human Endogenous Retroviral HERV-W Family Using PNA Strand Invasion into Double-Stranded DNA.

In the presented assay, we elaborated a method for distinguishing sequences that are genetically closely related to each other. This is particularly important in a situation where a fine balance of the allele abundance is a point of research interest. We developed a peptide nucleic acid (PNA) strand invasion technique for the differentiation between multiple sclerosis-associated retrovirus (MSRV) and ERVWE1 sequences, both molecularly similar, belonging to the human endogenous retrovirus HERV-W family. We have found that this method may support the PCR technique in screening for minor alleles which, in certain conditions, may be undetected by the standard PCR technique. We performed the analysis of different ERVWE1 and MSRV template mixtures ranging from 0 to 100% of ERVWE1 in the studied samples, finding the linear correlation between template composition and signal intensity of final reaction products. Using the PNA strand invasion assay, we were able to estimate the relative ERVWE1 expression level in human specimens such as U-87 MG, normal human astrocytes cell lines and placental tissue. The results remained in concordance with those obtained by semi-quantitative or quantitative PCR.

Perinatal outcome according to chorionicity in twins - a Polish multicenter study.

OBJECTIVES: The aim of the study was to analyze the perinatal outcome of twin gestations and estimate the influence of chorionicity on the outcome in a large cohort of twin pregnancies in Poland. MATERIAL AND METHODS: A retrospective analysis of 465 twin deliveries in 6 Polish centers in 2012 was conducted. Baseline characteristics, the course of pregnancy and labor, as well as the neonatal outcome were analyzed in the study group and according to chorionicity. RESULTS: A total of 356 twin pregnancies were dichorionic (DC group) (76.6%), and 109 were monochorionic (MC group) (23.4%). There were no differences in the occurrence of pregnancy complications according to chorionicity, except for IUGR of at least one fetus (MC 43.1% vs. DC 34.6%; p = 0.003). 66.5% of the women delivered preterm, significantly more in the MC group (78% vs. 62.9%; p = 0.004). Cesarean delivery was performed in 432 patients (92.9%). Mean neonatal birthweight was statistically lower in the MC group (2074 g vs. 2370 g; p < 0.001). Perinatal mortality of at least one twin was 4.3% (2.8% in the DC group vs. 9.2% in the MC group; p = 0.004). Neonatal complications, including NICU admission, respiratory disorders, and infections requiring antibiotic therapy, were significantly more often observed among the MC twins. CONCLUSIONS: The overall perinatal outcome in the presented subpopulation of Polish twins and its dependence on cho-rionicity is similar to the reports in the literature. Nevertheless, the rates of preterm and cesarean deliveries remain higher. It seems that proper counselling of pregnant women and education of obstetricians may result in reduction of these rates.

Late prematurity in twins: a Polish multicenter study.

The study aimed at investigating the impact of late prematurity (LPT) on neonatal outcome in twins and neonatal morbidity and mortality within LPT with regard to the completed weeks of gestation. The study was conducted in six tertiary obstetric departments from different provinces of Poland (Warsaw, Lublin, Poznan, Wroclaw, Bytom). It included 465 twin deliveries in the above centers in 2012. A comparative analysis of maternal factors, the course of pregnancy and delivery and neonatal outcome between LPT (34 + 0-36 + 6 weeks of gestation) and term groups (completed 37 weeks) was performed. The neonatal outcome included short-term morbidities. The analysis of neonatal complication rates according to completed gestational weeks was carried out. Out of 465 twin deliveries 213 (44.8%) were LPT and 156 (33.55%) were term. There were no neonatal deaths among LPT and term twins. One-third of LPT newborns suffered from respiratory disorders or required antibiotics, 40% had jaundice requiring phototherapy, and 30% were admitted to NICU. The analysis of neonatal morbidity with regard to each gestational week at delivery showed that most analyzed complications occurred less frequently with the advancing gestational age, especially respiratory disorders and NICU admissions. The only two factors with significant influence on neonatal morbidity rate were neonatal birth weight (OR = 0.43, 95% CI = 0.2-0.9, p = .02) and gestational age at delivery (OR = 0.62, 95% CI = 0.5-0.8, p < .01). LPT have a higher risk of neonatal morbidity than term twins. Gestational age and neonatal birth weight seem to play a crucial role in neonatal outcome in twins.

[Effects of the cigarette smoking on the newborn clinical parametrs and the accumulation of cadmium and lead in the placenta of women from Upper Silesia]. / Wplyw palenia papierosów na parametry noworodka oraz na kumulacje kadmu i olowiu w lozysku kobiet z Górnego Slaska.

AIM: The aim of the study was to determine the content of cadmium and lead in different parts of the placenta and fetal membranes of women who were exposed to cigarette smoke. The correlation between the two chemical elements and the impact of the Cd and Pb accumulation on newborn parameters were established. MATERIALS AND METHODS: The study material was collected immediately after delivery from 40 patients of the Department of Obstetrics and Gynecology Katowice. The marginal and central parts of the placenta and fetal membranes (amnion) were taken. The women were divided into two groups: smokers and non-smokers. Metal concentration in placenta was determined by flame atomic absorption spectrometry (FAAS). Bioethical Commission approved of the study RESULTS: In both studied groups, smokers and non-smokers, the presence of cadmium and lead was detected. Smokers turned out to have accumulated more of the investigated heavy metals in the placenta and fetal membranes. In the analyzed groups of women of smokers and non-smokers, differences in the content of the studied metals were found, but they were not statistically significant. Differences in newborn parameters in the two groups of women occurred, but again they lacked statistical significance. The level of lead increases along with the increase in the amount of cadmium, which proves the existence of a statistically significant correlation between them (p = 0.000). CONCLUSIONS: The number of smoked cigarettes increases cadmium content in maternal placenta and fetal membranes. No significant differences in newborn parameters of either smoker or non-smokers were observed, which may indicate women's adaptation to the environment containing cigarette smoke. The placenta and fetal membranes are biomarkers of the level of toxic exposure for the developing baby.

Mercury concentrations in human placenta, umbilical cord, cord blood and amniotic fluid and their relations with body parameters of newborns.

Studies were conducted on samples taken from giving birth women (n = 40) living in Poland, representing three age groups: 19-25, 26-30 and 31-38 years old. Mercury concentrations were measured with CV-AAS in placenta, umbilical cord, cord blood and amniotic fluid. The placentas weight did not exceed the 750 g value and was heavier than 310 g. Mean values of Hg concentrations in blood, placenta and umbilical cord were similar (c.a. 9 µg/g). High levels of mercury were noted in cord blood which in 75% of all observations exceeded (up to 17 µg/L) the safe dose set by US EPA (5.8 µg/L). No statistically significant differences in medium level of Hg in all the studied tissues among age groups of women were observed. Positive correlations between Hg concentrations in placenta and umbilical cord and cord blood were revealed as well as some negative ones between mercury concentrations and pregnancy parameters.

ADRB1 as a potential target for gene therapy of pregnancy induced hypertension and gestational diabetes mellitus.

Pregnancy-induced hypertension (PIH) and gestational diabetes mellitus (GDM) do not cause any problems with recognition; however, their pathophysiologies are still not explained. Yet many authors suggest that adrenergic ß-1 receptor (ADRB1) plays a crucial role. The aim of this study was to evaluate the transcription activity of ADRB1 by using real-time polymerase chain reaction (PCR) in placenta from normal pregnancies and from PIH and GDM. Obtained findings demonstrated a significant increase in ADRB1 mRNA expression in the examined groups in comparison to the control (p = 0.03). Our data indicate a potential perspective of ADRB1 suppression gene therapy in the treatment of PIH and GDM.

[Fetal ventriculomegaly secondary to germinal matrix hemorrhage--case report]. / Wentrikulomegalia w nastepstwie przebytego krwawienia do istoty zarodkowej mózgu plodu--opis przypadku.

A germinal matrix is an area of the fetal brain where neuroblasts proliferate till the 20th week of gestation after which their migration takes place. Germinal matrix hemorrhage (GMH) is a frequent cause of fetal ventriculomegaly or hydrocephalus, yet the pathophysiological conditions of GMH remain unclear. A case of fetal ventriculomegaly resulting from GMH has been reported in the article. A prenatal ultrasonographic examination (US) carried out in the 30th week of gestation revealed enlarged ventricles. Utero magnetic resonance imaging (MRI) performed in the 34th week of gestation demonstrated asymetric ventriculomegaly and the changes suggesting the occurance of GMH and ependymal intraventricular hemorrhage. Fetal ventriculomegaly and hydrocephalus are commonly detected by ultrasonographic examination, but it is often difficult to determine the causes of the diseases by US. In these cases MRI allows to establish the final diagnosis.

The polymorphism in insulin receptor substrate-1 gene and birth weight in neonates at term.

BACKGROUND: The mutation of the IRS-1 gene is one of the genetic risk factors which, it is speculated, is associated with insulin resistance or predisposition to type 2 diabetes. The aim of our study was to evaluate the association between the Gly972Arg polymorphism in the IRS-1 gene and birth weight in newborn children with adequate gestational age. MATERIAL AND METHODS: 100 newborn children with adequate gestational age (38-42 weeks), whose mother had no disorders during pregnancy, were studied. Genomic DNA was extracted from umbilical cord blood leukocytes, and Gly972Arg polymorphism in the IRS-1 gene was genotyped using the PCR-based method. RESULTS: Birth weight was significantly lower in the newborn with the IRS-1 Gly972Arg polymorphism compared with a control group (3161.75 +/- 380.86 g vs. 3427.92 +/- 468.86 g). Body length and head circumference at birth were also lower in the neonates with that polymorphism (54.38 +/- 3.13 cm vs. 52.69 +/- 2.91 cm, and 34.08 +/- 1.47 vs. 33.63 +/- 0.81, respectively). CONCLUSIONS: The results suggest that the Gly972Arg genotype is associated with lower birth weight, body length and head circumference in neonates with adequate gestational age.

Decreased human milk concentration of epidermal growth factor after preterm delivery of intrauterine growth-restricted newborns.

BACKGROUND AND AIM: The aim of the study was to compare epidermal growth factor (EGF) concentration in 81 colostrum samples collected from mothers of newborns in the following growth categories: preterm appropriate for gestational age (AGA), preterm small for gestational age (SGA), and full term (FT). RESULTS: Significantly higher concentrations of EGF were found in the colostrum of mothers who delivered premature AGA infants at less than 32 weeks of gestation compared with mothers who delivered premature SGA babies at the same gestational age. CONCLUSIONS: We concluded that the maternal compensatory mechanism accelerating the development of immature breast-fed infants may be disturbed when gestation is complicated by intrauterine growth retardation.

[Risk of fetal cytomegaly virus infection depending on number of CMV genome in mother's blood and amniotic fluid]. / Ryzyko zakazenia plodu wirusem cytomegalii w zaleznosci od ilosci kopii genomu CMV w krwi matki i plynie owodniowym.

The cytomegalovirus infection is most common causes of intrauterine infection of the fetus. Using of serologic diagnostic methods the kind of infection is unknown. The aim of the study was assessment of the risk of CMV infection depending of genome account in mother's blood and I amniotic fluid. The study was performed in choosen pregnancies, in which we expected cytomegalovirus infection using serological criteria. In prenatal diagnostic CMV infection using QPCR, the best material is amniotic fluid. Mother's blood assessment of CMV genome count does not make growth diagnostic possibility of the assesement of transmission the infection from mother to fetus.

[Diagnostic difficulties of Toxoplasma gondii infection in pregnant women. Is it possible to explain doubts by polymerase chain reaction?]. / Trudnosci w diagnostyce zarazenia ciezarnych Toxoplasma gondii. Czy metoda polimerazowej reakcji lancuchowej moze wyjasnic watpliwosci?

OBJECTIVES: Toxoplasma gondii infection during pregnancy is still a difficult problem in the contemporary perinatology. Difficulties met during interpretation of serological tests carried out in pregnant patients to detect Toxoplasmosis implies more and more frequent use of the Polymerase Chain Reaction (PCR). DESIGN: To evaluate the dependence between serological tests and quantity of the Toxoplasma gondii genomes in mothers' blood and amniotic fluid or neonatal blood, the quantitative PCR (q-PCR) method was applied. MATERIALS AND METHODS: The analysis was performed in 81 pregnant women. Maternal blood, amniotic fluid and newborns' umbilical blood samples were evaluated for the presence of Toxoplasma gondii DNA. IgG and IgM Toxoplasma gondii antibodies were evaluated by the ELISA method. RESULTS: High seroprevalence (51.9%) of the Toxoplasma gondii was confirmed. Toxoplasma gondii genetic material in blood and/or amniotic fluid was found in 33 patients. It was stated that quantity of the protozoa and anti-IgM presence in mothers' blood are the factors influencing significantly the Toxoplasma gondii manifestation in amniotic fluid. CONCLUSION: High suitability of PCR in diagnosis of Toxoplasmosis during pregnancy and vertical transmission was confirmed.