Neonatal Encephalopathy Group Research Articles

MicroRNAs as biomarkers of brain injury in neonatal encephalopathy: an observational cohort study

Neonatal Encephalopathy (NE) is a major cause of lifelong disability and neurological complications in affected infants. Identifying novel diagnostic biomarkers in this population may assist in predicting MRI injury and differentiate neonates with NE from those with low-cord pH or healthy neonates and may help clinicians make real-time decisions. To compare the microRNA (miRNA) profiles between neonates with NE, healthy controls, and neonates with low cord pH. Moreover, miRNA concentrations were compared to brain injury severity in neonates with NE. This is a retrospective analysis of miRNA profiles from select samples in the biorepository and data registry at the University of Florida Health Gainesville. The Firefly miRNA assay was used to screen a total of 65 neurological miRNA targets in neonates with NE (n = 36), low cord pH (n = 18) and healthy controls (n = 37). Multivariate statistical techniques, including principal component analysis and orthogonal partial least squares discriminant analysis, and miRNA Enrichment Analysis and Annotation were used to identify miRNA markers and their pathobiological relevance. A set of 10 highly influential miRNAs were identified, which were significantly upregulated in the NE group compared to healthy controls. Of these, miR-323a-3p and mir-30e-5p displayed the highest fold change in expression levels. Moreover, miR-34c-5p, miR-491-5p, and miR-346 were significantly higher in the NE group compared to the low cord pH group. Furthermore, several miRNAs were identified that can differentiate between no/mild and moderate/severe injury in the NE group as measured by MRI. MiRNAs represent promising diagnostic and prognostic tools for improving the management of NE.

Plasma UCHL-1 as a Biomarker of Brain Injury in Hospitalized Foals With Neonatal Encephalopathy

A plasma biomarker such as ubiquitin carboxyl-terminal hydrolase L1 (UCHL-1) to distinguish neonatal encephalopathy (NE) from other disorders and provide prognostic information would be useful for equine practitioners. In this prospective study, plasma UCHL-1 was measured in 331 hospitalized foals ≤4 days of age. Clinical diagnoses of neonatal encephalopathy only (NE group, n = 77), sepsis only (Sepsis group, n = 34), concurrent sepsis and NE (NE+Sepsis group, n = 85), or neither sepsis nor NE (Other group, n = 101) were made by the attending veterinarian. Plasma UCHL-1 concentrations were measured by ELISA. Differences between clinical diagnoses groups were evaluated and receiver operator curve (ROC) analysis was performed to assess diagnostic and prognostic performance. Median admission UCHL-1 concentration was significantly higher for NE (18.22 ng/mL; 7.93–37.43) and NE+Sepsis (17.42 ng/mL; 7.67–36.24) groups than Other foals (7.77 ng/mL; 3.92–22.76). Admission UCHL-1 was significantly higher in nonsurvivors (16.66 ng/mL; 6.89–34.84) than survivors (10.27 ng/mL; 5.82–29.94). Overall diagnostic performance of admission UCHL-1 concentration for NE diagnosis was determined (AUC 0.61; 95% confidence interval [CI] = 0.55–0.68); sensitivity and specificity for predicting NE were 73% and 49% respectively. Overall prognostic performance of time to lowest UCHL-1 concentration for predicting nonsurvival was determined (AUC 0.72; 95% CI = 0.65–0.79); sensitivity and specificity were 86% and 43% respectively. In this foal population, differences in plasma UCHL-1 concentrations were observed between foals with NE or NE with sepsis, and other diagnoses. The diagnostic and prognostic value of admission UCHL-1 concentration was limited.

Altered Cytokine Endotoxin Responses in Neonatal Encephalopathy Predict MRI Outcomes.

Background: Neonatal encephalopathy (NE) is associated with adverse neurodevelopmental outcome and is linked with systemic inflammation. Pro-inflammatory and anti-inflammatory cytokines are known to play a role in the pathology of NE by activating innate immune cells.Methods: Eighty-seven infants were enrolled including 53 infants with NE of whom 52 received therapeutic hypothermia (TH) and 34 term infant healthy controls (TC). Whole blood sampling was performed in the first 4 days of life, and a 14-spot ELISA Multiplex Cytokine Array was carried out on baseline samples or after stimulation with lipopolysaccharide (LPS) as an additional inflammatory stimulus. The cytokine medians were examined for differences between infants with NE and healthy TC; and then short-term outcomes of Sarnat stage, seizures, and MRI brain were examined within the NE group. The potential of LPS stimulation to predict abnormal MRI was explored using receiver operating characteristic (ROC) curves.Results: At baseline, infants with NE had significantly higher levels of erythropoietin (Epo), interleukin (IL)-6, and IL-1ra and significantly lower vascular endothelial growth factor (VEGF) than had controls. All cytokines were increased after LPS stimulation in infants with NE with an excessive Epo and IL-1ra response than in controls. Infants with NE had lower IL-8, IL-2, IL-6, tumor necrosis factor (TNF)-α, granulocyte-macrophage colony-stimulating factor (GM-CSF), VEGF, and interferon (IFN)-γ than controls had following LPS. GM-CSF and IFN-γ, IL-1β, IL-1ra, and VEGF were higher on days 1–2 in NE infants with abnormal neuroimaging. GM-CSF, IFN-γ, and TNF-α levels with LPS stimulation were different upon stimulation between normal and abnormal neuroimaging. TNF-α is the only strong cytokine predictor both pre- and post-LPS stimulation of abnormal brain imaging.Conclusions: Altered cytokine responses are found in infants with NE vs. controls, and more significant differences are unmasked by the additional stimulus of LPS, which potentially improves the predictive power of these cytokines for the detection of abnormal MRIs. Infants with NE undergoing TH demonstrate both trained immunity and tolerance, and understanding these responses will facilitate adjunctive immunomodulatory treatments.

Cytokine dysregulation persists in childhood post Neonatal Encephalopathy

BackgroundCytokines are possible mediators of neuroinflammation and associated with adverse outcome in neonatal encephalopathy (NE). Our aim was to explore cytokine response in children with Neonatal Encephalopathy (NE) at school age compared to age-matched controls.MethodFollow up at school age, children who had NE and age-matched controls were assessed for their cytokine responses and neurodevelopment outcome. Pro- and anti-inflammatory cytokines in the serum, [Interleukin (IL)-1α, IL-1β, IL-2, IL-6, IL-8, IL-18, Tumor necrosis factor (TNF)-α, TNF β, Interferon (IFN)-γ, granulocyte-macrophage colony-stimulating factor (GM-CSF), vascular endothelial growth factor (VEGF), erythropoietin (EPO), IL-10 & IL-1RA] were measured at baseline and in response to in vitro stimulation with lipopolysaccharide (LPS: endotoxin).ResultsGM-CSF, TNF-β, IL-2 IL-6 and IL-8 were significantly elevated at school age following NE (n = 40) compared to controls (n = 37). A rise in GM-CSF, IL-8, TNF-α, IL-1β, & IL-6 were seen in NE group following LPS stimulation. Relative LPS hypo-responsiveness was also noted in children with severe NE with IL-10, VEGF, EPO and TNF-β. Elevated TNF-β was associated with low gross motor scores on assessment at school age.ConclusionSchool-age children post-NE had significantly altered cytokine responses to endotoxin compared to controls. TNF-β was associated with adverse developmental outcomes. This suggests the inflammatory process may persist into childhood and a longer therapeutic window may be available for neuroprotection therapies.

Association of NOS1 gene polymorphisms with cerebral palsy in a Han Chinese population: a case-control study

BackgroundCerebral palsy (CP) is the leading cause of motor disability in children; however, its pathogenesis is unknown in most cases. Growing evidence suggests that Nitric oxide synthase 1 (NOS1) is involved in neural development and neurologic diseases. The purpose of this study was to determine whether genetic variants of NOS1 contribute to CP susceptibility in a Han Chinese population.MethodsA case-control study involving 652 CP patients and 636 healthy controls was conducted. Six SNPs in the NOS1 gene (rs3782219, rs6490121, rs2293054, rs10774909, rs3741475, and rs2682826) were selected, and the MassARRAY typing technique was applied for genotyping. Data analysis was conducted using SHEsis online software, and multiple test corrections were performed using SNPSpD online software.ResultsThere were no significant differences in genotype and allele frequencies between patients and controls for the SNPs except rs6490121, which deviated from Hardy-Weinberg equilibrium and was excluded from further analyses. Subgroup analysis revealed differences in genotype frequencies between the CP with neonatal encephalopathy group (CP + NE) and control group for rs10774909, rs3741475, and rs2682826 (after SNPSpD correction, p = 0.004, 0.012, and 0.002, respectively). The T allele of NOS1 SNP rs3782219 was negatively associated with spastic quadriplegia (OR = 0.742, 95% CI = 0.600–0.918, after SNPSpD correction, p = 0.023). There were no differences in allele or genotype frequencies between CP subgroups and controls for the other genetic polymorphisms.ConclusionsNOS1 is associated with CP + NE and spastic quadriplegia, suggesting that NOS1 is likely involved in the pathogenesis of CP and that it is a potential therapeutic target for treatment of cerebral injury.

An evaluation of fetal heart rate characteristics associated with neonatal encephalopathy: a case‐control study

We sought to identify fetal heart rate (FHR) characteristics that are associated with neonatal encephalopathy (NE). Retrospective case-control study. A single medical centre in Shanghai, China, 2006-2015. Women delivering a singleton, non-anomalous infant at ≥36 weeks' gestation diagnosed with NE (cases, n = 109) were compared with a group of women with unaffected infants (controls, n = 233). Two physicians blinded to the outcome independently reviewed FHR tracings during the last 30 minutes of tracing prior to delivery. FHR characteristics were compared in the two groups and multivariable logistic regression was used to adjust for confounding. Adjusted odds ratio (aOR) and 95% confidence interval (CI) for the presence of specific FHR categories and characteristics. Category II FHR tracings were observed in 89% of women prior to delivery and were not independently associated with NE. Notably, a category III FHR was observed in 17.4% of women in the NE group compared with 0.9% of women in the control group (aOR 44.99, 95% CI 7.23-279.97). Bradycardia, minimal/absent variability, late decelerations and prolonged decelerations were independently associated with NE, whereas accelerations were protective. Similar findings were found when the cases were limited to NE with arterial cord pH <7.1 and in a subgroup analysis of women with category II tracings. Category III tracings, while infrequent, are not uncommon prior to delivery among fetuses who develop NE. In contrast, most FHR tracings are category II prior to delivery; however, individual FHR characteristics within this category are associated with NE. This research was supported by the Interdisciplinary Programme of Shanghai Jiao Tong University. Category III tracings are not uncommon prior to delivery among fetuses who develop neonatal encephalopathy.

Might Bilirubin Serve as a Natural Antioxidant in Response to Neonatal Encephalopathy?

Neonatal asphyxia is often associated with hepatic injury. We hypothesized that this might lead to increased bilirubin concentrations. Term neonates admitted between January 2015 and April 2017 who remained hospitalized for ≥ 4 days and who had serial serum bilirubin concentrations recorded were divided into those with neonatal encephalopathy (NE) and controls. Serial serum bilirubin concentrations during the first days of life were compared between groups. Twenty-nine neonates with NE and 84 age-matched controls were identified. Mean total serum bilirubin concentrations of NE babies were significantly lower than those controls throughout the first days of life. At 96 hours of age, NE serum bilirubin concentrations were 4.5 (3.2, 5.8) versus controls of 10.5 (9.4, 11.5) mg/dL (p < 0.0001). The mean area under the curve (AUC) for the NE group was 268 (215, 321) versus 663 (608, 718), p < 0.0001, for the control group. All of the NE babies remained below the 40th percentile of the Bhutani curve and none required phototherapy. Contrary to our hypothesis, bilirubin concentrations in NE infants are significantly lower than expected during the first 4 days postnatally. We speculate that, under conditions of severe oxidative stress, bilirubin is consumed as an antioxidant.

ERP evidence of preserved early memory function in term infants with neonatal encephalopathy following therapeutic hypothermia.

Neonatal encephalopathy (NE) carries high risk for neurodevelopmental impairments. Therapeutic hypothermia (TH) reduces this risk, particularly for moderate encephalopathy (ME). Nevertheless, these infants often have subtle functional deficits, including abnormal memory function. Detection of deficits at the earliest possible time-point would allow for intervention during a period of maximal brain plasticity. Recognition memory function in 22 infants with NE treated with TH was compared to 23 healthy controls using event-related potentials (ERPs) at 2 wk of age. ERPs were recorded to mother's voice alternating with a stranger's voice to assess attentional responses (P2), novelty detection (slow wave), and discrimination between familiar and novel (difference wave). Development was tested at 12 mo using the Bayley Scales of Infant Development, Third Edition (BSID-III). The NE group showed similar ERP components and BSID-III scores to controls. However, infants with NE showed discrimination at midline leads (P = 0.01), whereas controls showed discrimination in the left hemisphere (P = 0.05). Normal MRI (P = 0.05) and seizure-free electroencephalogram (EEG) (P = 0.04) correlated positively with outcomes. Infants with NE have preserved recognition memory function after TH. The spatially different recognition memory processing after early brain injury may represent compensatory changes in the brain circuitry and reflect a benefit of TH.

G122(P) Melatonin and immune cell responses in neonatal encephalopathy

Introduction Infection and inflammation can be antecedents of Neonatal Encephalopathy (NE) and increase the risk of neurological sequelae. Melatonin is a potent immunomodulator and antioxidant (1) and may alter the systemic inflammatory response in NE (2). Aim To investigate the in vitro effect of melatoninon whole blood reactive oxygen intermediates (ROI), CD11b and Toll-like receptor (TLR)-4 in neutrophils and monocytes from infants with NE receiving therapeutic hypothermia (TH) versus healthy neonatal controls in the first week of life. Methods Infants with NE were recruited and their demographics details, grade of NE, MRI results, outcome and placental histology were recorded. Whole blood was taken on Day 1,3 and 7 of life (NE group) and day 1(Controls) and flow cytometry used to assess TLR4, CD11b and ROI in both monocytes and neutrophils in the presence of Lipopolysaccharide (LPS) and/or melatonin. Ethics was received from ethics committee at National Maternity Hospital. Results LPS–induced ROI production was significantly increased in both neutrophils and monocytes (p = 0.03) in NE versus controls (n = 6) on day 1 of life. On day 7 of life, following TH, LPS-induced CD11b upregulation was significantly decreased by melatonin in vitro in neonates with NE (n = 7). There was no difference in TLR4 expression in NE and controls. Conclusion Melatonin decreases the production of CD11b in neutrophils, which is a marker of neutrophil activation and migration and may ameliorate the augmented systemic inflammatory response seen in infants with NE.

1085 Cardiovascular Dysfunction in Infants with Neonatal Encephalopathy in the 1st Week of Life

Background and AimsPerinatal asphyxia may result in transient myocardial ischameia, confirmed by elevated Troponin T levels. Gold standard echocardiographic measures of contractility (ejection and shortening fraction) may not pick up...

The intrapartum CTG prior to neonatal encephalopathy at term: a case-control study.

To compare cardiotocograph (CTG) records during labour in cases of neonatal encephalopathy and matched controls. Case-control study. Metropolitan area of Perth, Western Australia. Term deliveries complicated by neonatal encephalopathy and controls matched for sex, hospital, time of birth, day of week of birth and maternal health insurance. Low fetal heart rate (FHR) variability, FHR accelerations, late decelerations, total Kreb's score and FIGO classification of CTG records. The neonatal encephalopathy group had significantly more abnormal CTG records (89%) classified according to FIGO, although 52% of control CTG records were also abnormal. CTG records from cases developed significant differences in terms of absence of FHR accelerations and low FHR variability, but not late decelerations, prior to delivery. Given the low incidence of neonatal encephalopathy in this study (7 per 1000) the predictive value of an abnormal CTG record is clinically unhelpful. However, the changes in the FHR in such cases suggest a greater disturbance of fetal (rest-activity) behaviour during labour.

Cerebral palsy and neonatal encephalopathy.

A retrospective cohort study was carried out to test the hypothesis that children born at term with cerebral palsy with signs of neurological dysfunction preceded by depression at birth (termed neonatal encephalopathy) differ from those without such signs in the frequency of antenatal and perinatal factors, and in the severity and characteristics of their impairment and disability. The study was carried out in the area covered by Oxford Regional Health Authority. Antenatal, intrapartum, neonatal factors, and the later clinical status of the two groups of children were used as the main outcome measures. Although most maternal and antenatal characteristics were similar in the two groups, the mothers of children with a history of neonatal encephalopathy were more likely to be primigravidae (odds ratio (OR) 2.0; 95% confidence interval (CI) 1.0 to 4.3) and to have a pregnancy of greater than 41 weeks' gestation (OR 3.5; 95% CI 1.0 to 12.1). Intrapartum complications were more frequent in the neonatal encephalopathy group: meconium staining of the amniotic fluid (OR 3.5; 95% CI 1.5 to 7.8), an ominous first stage cardiotocograph (OR 10.2; 95% CI 2.9 to 36.4), with a longer median duration of abnormality (200 v 48 minutes). At 5 years of age those with neonatal encephalopathy were more likely to have developed spastic quadriplegia (OR 4.8; 95% CI 2.2 to 10.5), to have visual impairment (OR 3.0; 95% CI 1.0 to 8.6), and to be non-walking (OR 4.0; 95% CI 1.8 to 8.8) than those without neonatal encephalopathy. Children with cerebral palsy who were born at term and have neonatal encephalopathy are more likely to have had signs of intrapartum asphyxia and are more likely to have a more severe form of cerebral palsy than those without a history of neonatal encephalopathy. Although this group represents only one in 10 of all cases of cerebral palsy, some of these may be obstetrically preventable.