Detection of pathogens associated with early-onset neonatal sepsis in cord blood at birth using quantitative PCR

Background: Early onset neonatal sepsis (EONS) typically begins prior to, during or soon after birth and may be rapidly fatal. There is paucity of data on the aetiology of EONS in sub-Saharan Africa due to limited diagnostic capacity in this region, despite the associated significant mortality and long-term neurological impairment. Methods: We compared pathogens detected in cord blood samples between neonates admitted to hospital with possible serious bacterial infection (pSBI) in the first 48 hours of life (cases) and neonates remaining well (controls). Cord blood was systematically collected at Kilifi County Hospital (KCH) from 2011-2016, and later tested for 21 bacterial, viral and protozoal targets using multiplex PCR via TaqMan Array Cards (TAC). Results: Among 603 cases (101 [17%] of whom died), 179 (30%) tested positive for ≥1 target and 37 (6.1%) tested positive for multiple targets. Klebsiella oxytoca, Escherichia coli/Shigella spp., Pseudomonas aeruginosa, and Streptococcus pyogenes were commonest. Among 300 controls, 79 (26%) tested positive for ≥1 target, 11 (3.7%) were positive for multiple targets, and K. oxytoca and P. aeruginosa were most common. Cumulative odds ratios across controls: cases (survived): cases (died) were E. coli/Shigella spp. 2.6 (95%CI 1.6-4.4); E. faecalis 4.0 (95%CI 1.1-15); S. agalactiae 4.5 (95%CI 1.6-13); Ureaplasma spp. 2.9 (95%CI 1.3-6.4); Enterovirus 9.1 (95%CI 2.3-37); and Plasmodium spp. 2.9 (95%CI 1.4-6.2). Excluding K. oxytoca and P. aeruginosa as likely contaminants, aetiology was attributed in 9.4% (95%CI 5.1-13) cases using TAC. Leading pathogen attributions by TAC were E. coli/Shigella spp. (3.5% (95%CI 1.7-5.3)) and Ureaplasma spp. (1.7% (95%CI 0.5-3.0)). Conclusions: Cord blood sample may be useful in describing EONS pathogens at birth, but more specific tests are needed for individual diagnosis. Careful sampling of cord blood using aseptic techniques is crucial to minimize contamination. In addition to culturable bacteria, Ureaplasma and Enterovirus were causes of EONS.

Iron metabolism disorder and oxidative damage levels in placenta are involved in preeclampsia

IntroductionTo explore the role of ferritin in placenta, serum and umbilical cord blood of pregnant women and the changes of oxidative stress injury as well as cell apoptosis in placenta in the pathogenesis of preeclampsia (PE).Material and methodsSixty pregnant women with severe PE were assigned into early-onset and late-onset PE group. Another 60 cases of normal late pregnant women with similar gestational weeks were divided into early-onset and late-onset control group. Maternal serum and fetal umbilical cord blood ferritin content was determined by automatic biochemical immunoassay system; mRNA expression levels of ferritin and ferritin heavy chain (FTH) were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction (RT-qPCR). Western Blot was used to detect the relative expression level of ferritin and apoptosis; the contents of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were detected by colorimetry.ResultsSerum uric acid (UA) and creatinine (Cr) levels of PE groups were significantly higher when compared to the controls. The serum ferritin levels in blood sample and umbilical cord blood sample were significantly higher relative to the controls. However, the mRNA and protein levels of ferritin levels in placenta samples were significantly lower compared with the controls. The placental cleaved caspase-3, Bcl-2 levels were significantly lower than the early onset PE group. The levels of GSH-Px and MDA in placenta were significantly higher.ConclusionsThese results may assist understanding the pathogenesis of PE and provide potential biomarkers for diagnosis of PE.

The link between cord blood IL-1β, TLR4, PGE2 and TAC values with neonatal diseases

Premature birth is an important cause of neonatal mortality and neonatal morbidity. Most premature births are known to be induced by cytokines released for different reasons. Inadequate congenital immune response in premature infants may contribute to increased susceptibility to infection. The aim of the study is to determine the IL-1β, TLR4, PGE2, and TAC profiles in cord blood with characteristics specific to pregnancy and the correlation with neonatal complications caused by premature birth. The study included 26 neonates, 11 girls and 15 boys, born from 24-42 weeks of gestation. Of these, 13 were term and 13 were preterm . For IL-β, PG-E2, TLR4 and TAC levels, 1 mL of cord blood sample was taken from preterm and term neonates. Data related to demographic data, clinical status of patients and outcomes were obtained from electronic medical records and files. Cytokine values obtained from premature neonates were statistically high in terms of TLR4, IL1 and PGE2 compared to term infants. The TRL4 and IL1 values for premature infants with necrotizing enterocolitis and retinopathy of prematurity were lower compared to those without NEC and ROP. In spite of negative correlations between TAC and the other three cytokines, a statistically significant correlation was not identified. TLR4, IL1 and PGE2 were negatively correlated with weight and gestational week, contrarily TAC measurements were positively correlated with weight and gestational week. Measurements of cytokine concentrations in cord blood are among important biomarkers showing degree of inflammation and may assist in predicting neonatal complications and play an effective role in development of specific treatments.

The Correlations between Cord Blood Leptin and Leptin Level at Six Months with Infant Growth

Background: Leptin plays an important role in regulating body weight, metabolism, and reproductive functions. Leptin affects metabolism by reducing nutrient intake and increasing energy expenditure which eventually also plays a role in infant growth.Objective: This study aims to determine the relationship between leptin levels and infant growth age 0-6 months.Methods: A prospective cohort study was done for six months on 38 infants, age 0-6 months, from breastfeeding mothers with normal pregnancies. The samples were taken twice, firstly when the infant was born using an umbilical cord blood sample, and secondly at the age of six months, using a vein blood sample. Serum leptin levels were measured using the ELISA method. Infant growth was assessed using WHO 2005’s z-scores.Results: A total of 50 babies were included in the study, 38 of them had been studied completely. Significant correlations were found between the mean of the umbilical cord and six months of age leptin levels (p <0.001), between delta leptin with WHZ and delta leptin with WAZ at six months of age (p = 0.002 and p = 0.003, respectively), and between leptin levels with WHZ (p<0.001) and leptin levels with WAZ (p = 0.004) at six months of age. Leptin levels at the age of six months are lower than umbilical cord blood leptin. Conclusion: The greater decrease of leptin level in the first six months is associated with better infant growth.

Correlation between Intrapartum Cardiotocogram findings and cord blood pH in term and preterm labors

Background: The purpose of intrapartum fetal monitoring by cardiotocograph (CTG) is to identify early signs of developing hypoxia so that appropriate action can be taken to improve the perinatal outcome. Although CTG findings are well known to monitor the progress of the labor due to the paucity of recommendations, there has always been a clinical dilemma as the term fetuses respond differently than a preterm fetus. However, umbilical cord blood pH can distinguish the infant at high risk for asphyxia and related sequel. Therefore, because of differences in fetal physiology in term and preterm fetuses, CTG findings vary, and hence the validity of CTG to determine fetal acidosis should be different. Aims and Objectives: This study aimed to correlate abnormal intrapartum CTG findings with umbilical cord blood pH in term and preterm labor and thus evaluate the success of CTG in predicting fetal acidosis during labor. Methods: The present study included 210 women in labor (70 preterm and 140 term) with abnormal intrapartum CTG that was classified as per 2015 revised International Federation of Gynecologists and Obstetrician (FIGO) guidelines. Immediately after delivery 2 ml Umbilical artery cord blood sample was taken in a pre-heparinized syringe for analysis, pH <=7.2 was taken as acidosis and pH >7.2 was taken as normal. The measured data were maternal general characteristics which included gravida status, associated comorbidities, method of induction and character of liquor, the intrapartum CTG tracings recorded the cord arterial blood pH and the neonatal characteristics such as APGAR score and neonatal outcome. Results: Data from 70 preterm labor was compared with 140 term labor. In this study, 20.9 % of the babies had acidosis. Suspicious CTG due to decreased variability were more common in the preterm group than in the term group (21.4% vs. 8.6% p<0.05). Positive predictive value (PPV) of abnormal CTG for fetal acidosis in the preterm group was found to be higher than that in term group, PPV of pathological CTG being even higher than suspicious CTG. Women with suspicious CTG had 82 % less risk of fetal acidosis as compared to pathological CTG. Women with Bradycardia had 5.9 times the risk of fetal acidosis as compared with normal and tachycardia. Conclusion: Abnormal CTG should be managed appropriately without any delay to prevent acidosis and cord blood pH should be done in all labors with abnormal CTG. However, our findings of a higher incidence of lower cord blood pH and suspicious CTG due to decreased variability alone, highlight the limitation of criteria currently used for interpretation of CTG in preterm labors.

Relationship between Maternal Hemoglobin Concentration to the Newborn Cord Blood Hemoglobin and Serum Concentration

Objective: To determine the mean cord blood hemoglobin and ferritin concentration in newborn of anemic mothers. Study Design: Cross sectional Place and Duration: Gynae & Obs department of Dow University Hospital, a tertiary care hospital in Karachi, Pakistan during the period from January, 2018 to July, 2019. Methodology: A total of 100 anemic pregnant women as per inclusion and exclusion criteria were included in this study. Cord blood sample (5ml) was obtained by sterile needle puncture immediately after cord clamping. 2ml will collected in EDTA bottle for full blood count analysis and 3ml collected in plain bottle for serum ferritin analysis. Final outcome was recorded on approved proforma. Results: The average age of the patients was 29.33±3.23 years. Mean cord blood hemoglobin and ferritin concentration in newborn of anemic mothers was 14.78±1.62 g/dl and 94.10±67.47 u/ml. Conclusion: We found that average hemoglobin of the women recruited in our study was 9.59±0.815 g/dl however we could find no correlation between mean cord blood hemoglobin and ferritin concentration in newborn of anemic mothers. Key Words: Cord blood Hemoglobin, Ferritin, Anemic mothers

Evaluation of SARS-CoV-2 Spike Protein Antibody Titers in Cord Blood after COVID-19 Vaccination during Pregnancy in Polish Healthcare Workers: Preliminary Results

Background: The coronavirus disease 2019 (COVID-19) pandemic has given rise to the need to develop a vaccine as quickly as possible. As pregnant women are at increased risk of contracting severe COVID-19, with higher mortality, it is essential to assess the safety of the vaccines administered during pregnancy. Methods: The aim of this study was to determine the titer of specific maternal and cord antibodies against severe acute respiratory syndrome coronavirus 2 S protein after antenatal vaccination. The secondary objective was to evaluate the ratio of the umbilical cord to the maternal antibody titers. Patients included in the study were enrolled after undergoing voluntary vaccination against COVID-19 during pregnancy at different weeks of gestation. All patients analyzed in our initial study were vaccinated with the BNT162b2 mRNA COVID-19 vaccine. Results: The results of the current study document high anti-S total IgG antibody titers in cord serum at birth in all mother–infant pairs analyzed. The mean umbilical cord blood sample IgG antibody titer anti-S protein was 1026.51 U/mL (±SD 769.25). The mean cord-to-maternal anti–S IgG antibody ratio was 1.28 (±SD 0.798). A significant positive correlation was observed between the week of gestation at which the first dose was administered and the week of gestation at which the second dose was administered, and the respective cord-to-maternal ratio (r = 0.48; p = 0.0029) for the first dose and (r = 0.39; p = 0.0102) for the second dose. Conclusions: To date, despite the prevalence of COVID-19 vaccination, there is a lack of conclusive evidence supporting the safety and efficacy of vaccination of pregnant women. Therefore, the results we present are complementary. Our study suggests that maternal immunization may provide neonatal protection through the transplacental transfer of antibodies. Of particular importance is the demonstration that antibody transfer is correlated with the time from vaccination to delivery, which may allow future determination of the optimal timing of COVID-19 vaccination in pregnant women.

Optimising the Timing of whooping cough Immunisation in MUMs: a randomised controlled trial investigating the timing of pertussis vaccination in pregnancy (OpTIMUM): a protocol paper

Background: Pertussis is a highly infectious respiratory illness caused by the bacteria Bordetella pertussis. A resurgence of pertussis, even in countries with good vaccine coverage, has led to an increase in infant deaths. In response to this, many countries have introduced pertussis vaccination in pregnancy. This strategy is effective at preventing infant disease, but there remains uncertainty about what gestational timing is best to ensure maximal protection of the infant. These uncertainties are the rationale for this randomised controlled trial and a sub-study investigating pertussis-specific antibody in breastmilk. Protocol: We will recruit 354 pregnant women and will randomise them to receive their pertussis vaccination in one of three gestational age windows: ≤23+6, 24-27+6 and 28-31+6 weeks of gestation. Vaccination will be with Boostrix-IPV® and participants will be asked to complete a symptom diary for seven days following vaccination. Blood sampling will be performed prior to vaccination, two weeks following vaccination and at the time of delivery. A cord blood sample will be collected at delivery and a blood sample collected from the infant 4-10 weeks after completion of the primary immunisations. Individuals participating in the breastmilk sub-study will provide a sample of colostrum within 48 hours of delivery and samples of breastmilk at two weeks and around five-six months. Blood samples will be analysed using enzyme linked immunosorbent assay (ELISA) techniques for pertussis toxin, filamentous haemagglutinin and pertactin. A subset of serum samples will also be analysed using a functional assay. Colostrum and breastmilk samples will be analysed using functional assays. Discussion: Although pertussis vaccination has been shown to be safe and effective in pregnancy there remains debate about the optimal timing for the administration during pregnancy. This study will investigate antibody responses in serum and breastmilk when vaccination is performed in three different time periods. Clinicaltrials.gov registration: NCT03908164 (09/04/2019)

The differential expression and regulatory networks of ceRNAs in umbilical cord blood sample of chromosome 22q11.2 deletion syndrome

Background: Chromosome 22q11.2 deletion (CH22qD) syndrome is the most common human deletion syndrome. Competing endogenous RNAs (ceRNAs) have miRNA binding sites that are capable of competitively binding miRNAs and inhibiting miRNA regulation of target genes. Results: We identified differently expressed miRNAs, circRNAs, lncRNAs and mRNAs of CH22qD, and we analysed the results by using GO analysis, KEGG pathway analysis and network regulation analysis. Conclusions: These analyses may predict the effects of chromosomal microdeletions.

Functional Comparison of Blood-Derived Human Neural Progenitor Cells

Induced pluripotent stem cell (iPSC)-derived neural progenitor cells (NPCs) are promising tools to model complex neurological or psychiatric diseases, including schizophrenia. Multiple studies have compared patient-derived and healthy control NPCs derived from iPSCs in order to investigate cellular phenotypes of this disease, although the establishment, stabilization, and directed differentiation of iPSC lines are rather expensive and time-demanding. However, interrupted reprogramming by omitting the stabilization of iPSCs may allow for the generation of a plastic stage of the cells and thus provide a shortcut to derive NPSCs directly from tissue samples. Here, we demonstrate a method to generate shortcut NPCs (sNPCs) from blood mononuclear cells and present a detailed comparison of these sNPCs with NPCs obtained from the same blood samples through stable iPSC clones and a subsequent neural differentiation (classical NPCs—cNPCs). Peripheral blood cells were obtained from a schizophrenia patient and his two healthy parents (a case–parent trio), while a further umbilical cord blood sample was obtained from the cord of a healthy new-born. The expression of stage-specific markers in sNPCs and cNPCs were compared both at the protein and RNA levels. We also performed functional tests to investigate Wnt and glutamate signaling and the oxidative stress, as these pathways have been suggested to play important roles in the pathophysiology of schizophrenia. We found similar responses in the two types of NPCs, suggesting that the shortcut procedure provides sNPCs, allowing an efficient screening of disease-related phenotypes.