Evidence for neurobehavioral risk phenotypes at birth.

Observations of newborn behavior provide clinicians and researchers with a first description of the neurobehavioral organization of the newborn that is largely independent of the postnatal environment. The Neonatal Network Neurobehavioral Scale (NNNS) was developed in 2004 to evaluate how prenatal exposure to substances such as cocaine is related to neurobehavioral outcomes. There are now 156 empirical articles published using the NNNS, which we review and summarize. Z-scores from published studies using the NNNS were compiled and aggregated supporting the replicability of three newborn neurobehavioral phenotypes: one typical and two that are predictive of later cognitive and behavioral delay; hyper- and hypo-dysregulated newborns. These phenotypes emerged from independent samples and research groups and were identified in a variety of populations, including infants with prenatal substance exposure, preterm infants, and healthy term infants. Our findings show that newborn neurobehavior can be measured in a reliable and valid manner and that certain behavioral phenotypes, identifiable at birth, can predict neurodevelopmental challenges. These findings have important clinical utility. Intervening early with infants exhibiting these risk phenotypes may prevent later neurodevelopmental delay. IMPACT: We reviewed all empirical studies published using the Neonatal Network Neurobehavioral Scale and found evidence for two replicable stress phenotypes that predict later behavioral outcomes. This study highlights the utility of the Neonatal Network Neurobehavioral Scale for early identification of newborn neurodevelopmental risk phenotypes. Early identification of neurodevelopmental risk, when neuroplasticity is high, may ultimately reduce the burden of subsequent neurobehavioral problems through early intervention.

Testing the mid-range model: Attachment in a high risk sample.

Infant attachment is a key predictor of later socioemotional functioning, but it is not clear how parental responsivity to infant expressive behavior is associated with attachment outcomes. A mid-range model of responsivity holds that both unresponsive and highly reactive parental behaviors lead to insecure and disorganized attachment. We examined the relationship between maternal (and infant) contingent responsivity and attachment in a high-risk sample. Participants were 625 infant-mother pairs from a longitudinal study of children with and without prenatal drug exposure and variable levels of associated social risks. Infant-mother pairs participated in the Face-to-Face/Still-Face paradigm (FFSF) at 4-months and in the Strange Situation Procedure (SSP) at 18-months. A model incorporating both linear and quadratic responsivity effects indicated that mothers who were either very high (reactive) or very low (unresponsive) in responsivity were more likely to have infants with disorganized attachment outcomes. While maternal responsivity was associated with attachment disorganization, no associations between maternal responsivity, and attachment security/insecurity were detected. Infant responsivity to mother was not associated with attachment outcomes. The findings suggest the importance of mid-range levels of maternal responsivity in the development of organized attachment among infants facing high levels of prenatal and social risk.

From Early Micro-Temporal Interaction Patterns to Child Cortisol Levels: Toward the Role of Interactive Reparation and Infant Attachment in a Longitudinal Study.

Parental mental disorders increase the risk for insecure attachment in children. However, the quality of caregiver-infant interaction plays a key role in the development of infant attachment. Dyadic interaction is frequently investigated via global scales which are too rough to uncover micro-temporal mechanisms. Prior research found that the latency to reparation of uncoordinated dyadic states is associated with infant behavioral and neuroendocrine regulation. We investigated the hypothesis that this interactive mechanism is critical in predicting secure vs. insecure attachment quality in infancy. We also assessed the predictive quality of infant attachment regarding neuroendocrine reactivity later in childhood. A subsample of N = 58 dyads (n = 22 mothers with anxiety disorders, n = 36 controls) from a larger study were analyzed. At 3-8 months postpartum, maternal anxiety disorders were diagnosed via a structured clinical interview as well as dyadic interaction during the Face-to-Face-Still-Face (FFSF) was observed and coded on a micro-temporal scale. Infant attachment quality was assessed with the strange situation paradigm at 12-24 months of age. In an overlapping subsample of N = 39 (n = 13 mothers with anxiety disorder; n = 26 controls), we assessed child cortisol reactivity at 5 to 6 years of age. Generalized linear modeling revealed that longer latencies to interactive reparation during the reunion episode of the FFSF as well as maternal diagnosis at 3-8 months of age predict insecure attachment in children aged 12-24 months. Cox regressions demonstrated that dyads with infants who developed insecure attachment at 12-24 months of age were 48% less likely to achieve an interactive reparation at 3-8 months of age. Mixed models revealed that compared to securely attached children, children who had developed an insecure attachment at 12-24 months of age had an increased cortisol reactivity at 5 to 6 years of age during free play. The results confirm the hypothesis that the development of attachment is affected by experienced micro-temporal interactive patterns besides diagnostic categories. They also showed that infants of mothers with postpartum anxiety disorders have a more than fivefold increased risk of developing an insecure attachment than the infants of the control group. Moreover, results imply that these patterns may influence neurohormonal regulation even in preschool aged children.

Maternal hair cortisol levels as a novel predictor of neonatal abstinence syndrome severity: A pilot feasibility study.

Neonatal abstinence syndrome (NAS) after in-utero opioid exposure remains a poorly understood condition with multiple factors contributing to severity. Exposure to maternal stress may be one contributing factor. Hair cortisol measurement represents a novel technique for assessing prenatal stress. In this pilot study, the association between maternal hair cortisol levels and NAS severity was examined in 70 postpartum women with opioid use disorder within 72 hr of delivery. Infants were monitored for NAS and treated according to institutional protocol. Forty-four (63%) of the infants were pharmacologically treated for NAS, with a mean length of hospital stay (LOS) for all infants of 14.2 (SD 9.0) days. The mean cortisol level in the mothers was 131.8 pg/mg (SD 124.7). In bivariate analysis, higher maternal hair cortisol levels were associated with shorter infant LOS (R = -.26, p = .03) and fewer infant opioid treatment days (R = -.28, p = .02). Results were no longer statistically significant in regression models after adjusting for maternal opioid and smoking. In conclusion, we demonstrated the feasibility of hair cortisol assaying within the first few days after delivery in mothers with opioid use disorder as a novel marker for NAS. The findings suggest that maternal stress may impact the severity of infant opioid withdrawal.

DNA methylation of NR3c1 in infancy: Associations between maternal caregiving and infant sex.

Caregivers play a critical role in scaffolding infant stress reactivity and regulation, but the mechanisms by which this scaffolding occurs is unclear. Animal models strongly suggest that epigenetic processes, such as DNA methylation, are sensitive to caregiving behaviors and, in turn, offspring stress reactivity. We examined the direct effects of caregiving behaviors on DNA methylation in infants and infant stress reactivity. Infants and mothers (N = 128) were assessed during a free play when infants were 5 months old. Maternal responsiveness and appropriate touch were coded. and infant buccal epithelial cells were sampled to assess for DNA methylation of the glucocorticoid receptor gene, NR3c1 exon 1F. Infant cortisol reactivity was assessed in response to the still-face paradigm. Greater levels of maternal responsiveness and appropriate touch were related to less DNA methylation of specific regions in NR3c1 exon 1F, but only for females. There was no association with maternal responsiveness and appropriate touch or DNA methylation of NR3c1 exon 1F on prestress cortisol or cortisol reactivity. Our results are discussed in relation to programming models that implicate maternal care as an important factor in programing infant stress reactivity.

Los cuidadores juegan un papel esencial en el andamiaje de la reactividad y regulación del estrés infantil pero los mecanismos por medio de los cuales aparece este andamiaje no están claros. Los modelos animales fuertemente sugieren que los procesos epigenéticos, tales como la metilación del ADN, son sensibles a los comportamientos de prestaciones de cuidado y por consiguiente a la reactividad al estrés por parte de los hijos. Examinamos los efectos directos que los comportamientos de prestaciones de cuidado tienen sobre la metilación de ADN en infantes y, por consiguiente, la reactividad del estrés infantil. Los infantes y sus madres (N = 128) fueron evaluados durante una sesión de juego libre cuando los infantes tenían 5 meses de edad. Se codificó la sensibilidad materna y la apropiada forma de tocar y se obtuvo muestra de las células epiteliales bucales del infante para analizar la metilación de ADN del gen receptor glucocorticoide, NR3c1, exón 1F. Se evaluó la reactividad del infante al cortisol como respuesta al paradigma de la cara quieta. Niveles mayores de sensibilidad materna y apropiada forma de tocar se relacionaron con menos metilación de ADN de regiones específicas en NR3c1 exón 1F, aunque sólo en las niñas. No se presentó ninguna asociación con la sensibilidad materna y la apropiada forma de tocar, o metilación de ADN de NR3c1 exón 1F en el cortisol pre-estrés o la reactividad del cortisol. Nuestros resultados se discuten en relación con modelos de programación que implican cuidado materno como un importante factor en la programación de la reactividad del estrés del infante.

Les personnes prenant soin des enfants jouent un rôle critique dans l'échafaudage de la réaction au stress du nourrisson et la régulation mais les mécanismes selon lesquels cet échafaudage se bâtit ne sont pas clairs. Les modèles animaux suggèrent fortement que des processus épigénétiques, comme la méthylation de l'ADN, sont sensibles au comportements de qui prend soin d'eux et en conséquence déclenchent un réaction au stress. Nous avons examiné les effets directs des comportements soignants sur la méthylation de l'ADN chez les bébés, en ensuite sur la réaction au stress du nourrisson. Des nourrissons et leurs mères (N = 128) ont été évalués au moyen d'un jeu libre quand les bébés avaient 5 mois d'âge. La réaction maternelle et le toucher approprié ont été codés et des cellules épithéliales buccales du bébé ont été prélevées afin d'évaluer la méthylation de l'ADN du gène récepteur glucocorticoïde, le NR3c1 exon 1F. La réaction du cortisol du bébé a été évaluée en réponse au paradigme du visage immuable. Des niveaux plus élevés de réaction maternelle et de toucher approprié étaient liés à une méthylation de l'ADN des régions spécifiques de NR3c1 exon 1F moindre, mais seulement chez les filles. On n'a trouvé aucun lien avec la réaction maternelle et le toucher approprié ou de méthylation NR3c1 exon 1F de l'ADN sur le cortisol pré-test ou de réaction du cortisol. Nos résultats sont discutés en relation aux modèles de programme qui impliquent que le soin maternel en tant que facteur important dans la programmation de la réaction au stress du bébé.

Predicting Patterns of Regulatory Behavior in the Still-Face Paradigm at 3 Months.

The current study addressed two aims: (1) to describe different patterns of infant regulatory behavior during the Face-to-Face Still-Face (FFSF) paradigm at 3 months of age and (2) to identify specific, independent predictors of these patterns from an a priori set of demographic, infant (e.g., temperament), and maternal (e.g., sensitivity) variables. Analyses were based on data collected for 121 mother-infant dyads assessed longitudinally in the newborn period and again at 3 months. In the newborn period, infants' neurobehavior was evaluated using the Neonatal Behavioral Assessment Scale (NBAS) and mothers reported on their caregiving confidence and their newborns' irritability and alertness. At 3 months, mothers reported on their infant's temperament, and mother-infant interactions were videotaped during free play and the FFSF. Three patterns of infant regulatory behavior were observed. The most common was a Social-Positive Oriented Pattern, followed by a Distressed-Inconsolable Pattern, and a Self-Comfort Oriented Pattern. Results of multinomial logistic regression indicated that categorical assignment was not associated with demographic or infant characteristics, but rather with dyadic regulatory processes in which maternal reparatory sensitivity played a crucial role.

Social Stress-Related Epigenetic Changes Associated With Increased Heart Rate Variability in Infants.

Early life stress can result in persistent alterations of an individual's stress regulation through epigenetic modifications. Epigenetic alteration of the NR3C1 gene is associated with changes in the stress response system during infancy as measured by cortisol reactivity. Although autonomic nervous system (ANS) reactivity is a key component of the stress response, we have a limited understanding of the effects of NR3C1 DNA methylation on ANS reactivity. To examine this relation, ANS stress responses of term, 4-5-month-old healthy infants were elicited using the face-to-face still-face paradigm, which involved five, 2-min episodes. Two of these episodes were the "still-face" in which the mother was non-responsive to her infant. EKG was acquired continuously and analyzed in 30 s-intervals. Cheek swabs were collected, and DNA was extracted from buccal cells. Respiratory sinus arrhythmia (RSA) was measured as heart rate variability (HRV). Mean HRV was calculated for each 30-s "face to face" episode. DNA methylation of NR3C1 was calculated using bisulfite pyrosequencing. Percent DNA methylation was computed for each of the 13 NR3C1 CpG sites. The relations between mean HRV for each "face to face" episode and percent DNA methylation was examined averaged over CpG sites 1-6 and 7-13 and at each individual CpG site. Higher HRV at baseline, first reunion, and second still-face was related to greater methylation of NR3C1 CpG sites 1-6. Higher HRV at the second reunion was related to greater methylation of NR3C1 CpG sites 12 and 13. These data provide evidence that increased methylation of NR3C1 at CpG sites 12 and 13 are associated with increased activation of parasympathetic pathways as represented by increased HRV.

Robust stability and physiological correlates of infants' patterns of regulatory behavior in the still-face paradigm at 3 and 9 months.

This study examined the stability of three patterns of infant regulatory behavior identified in the face-to-face still-face (FFSF) paradigm at 3 and 9 months-social-positive oriented, distressed-inconsolable, and self-comfort oriented-and whether variations in infants' heart-rate were correlated with them. Although some studies have examined the stability of discrete infant behaviors, none have investigated the stability of early regulatory patterns across FFSF episodes over time. Healthy full-term infants and their mothers (N = 112) were videotaped in the FFSF when infants were 3 and 9 months old. Infants' regulatory patterns were scored with the Coding System for Regulatory Patterns in the FFSF. Infants' heart-rate level during each episode of the FFSF was also assessed. The social-positive-oriented pattern was the most prevalent at both ages. Cross-tabulation analysis showed a robust stability (Cohen's κ = .72) of the regulatory patterns from 3 to 9 months. The heart-rate level of infants with a social-positive-oriented pattern at 3 and 9 months showed recovery to baseline levels following the still-face. In contrast, the heart-rate level of infants with a distressed-inconsolable pattern at 9 months increased from the still-face to the reunion episode, whereas the heart-rate level of infants with a self-comfort-oriented pattern at 9 months did not change from the still-face to the reunion episodes. These results suggest that infants exhibit distinct organized regulatory patterns as early as 3 months that are stable over a 6-month interval and associated with variations in infants' physiological responses across FFSF episodes at both ages. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Epigenetic Programming by Maternal Behavior in the Human Infant.

: media-1vid110.1542/5804912859001PEDS-VA_2017-1890Video Abstract OBJECTIVES: We sought to determine if variations in maternal care alter DNA methylation in term, healthy, 5-month-old infants. This work was based on landmark studies in animal models demonstrating that nurturing care by dams would alter their newborns' stress responses through epigenetic mechanisms. We used breastfeeding as a proxy for animal maternal behavior. We hypothesized alterations in DNA methylation of the glucocorticoid receptor gene and less hypothalamic stress response in infants of mothers who breastfed their infants versus infants of mothers who did not breastfeed. METHODS: A cohort study of term, healthy infants and their mothers who did (n = 21) or did not (n = 21) breastfeed for the first 5 months was used in this analysis. Cortisol stress reactivity was measured in infant saliva by using a mother-infant interaction procedure and DNA methylation of an important regulatory region of the glucocorticoid receptor gene. Changes in DNA methylation of this gene in humans were compared to homologous regions of the rat gene. DNA samples were prepared from cheek swabs and subjected to quantitative analysis of the extent of methylation by using sensitive sequencing techniques. RESULTS: Breastfeeding was associated with decreased DNA methylation of the glucocorticoid receptor promoter and decreased cortisol reactivity in 5-month-old infants. Decreased DNA methylation occurred in the promoter region involved in regulation of the hypothalamic-pituitary-adrenal and immune system responses. CONCLUSIONS: Maternal care in humans may impact the hypothalamic-pituitary-adrenal stress response through behavioral programming and manifest as offspring epigenetic change. These results explain, in part, some of the positive effects observed in children who are breastfed.

Neonatal Behavioral Assessment Scale (NBAS): Confirmatory factor analysis of the six behavioral clusters.

The Neonatal Behavioral Assessment Scale (NBAS) is a widely used in the neurobehavioral assessment of neonates in clinical practice and research. Lester's data reduction system for the NBAS items is the most often used in research, but the few factor analytic studies carried out with it leave gaps in its validation. The current study aimed to test and compare (a) the factorial structure of the Lester's data reduction system for the NBAS and (b) an alternative data reduction system, slightly modified from Lester's system. The NBAS was administered to 196 healthy Portuguese full-term infants (51% male) in the first 72 h of life (M = 43.63 h). Construct validity of the data reduction systems was tested through confirmatory factor analysis (CFA). Lester's original system was compared to three alternative models, two of which included a revision of the scoring rules for three items and the exclusion of five items. The CFA generally supported the six-factor structure. However, fit indices for Lester's original model were only fair. An alternative, revised model with a second-order factor - Self-Organizing System - demonstrated a better fit. The results provide evidence to support a modified form of Lester's six behavioral clusters as a data reduction model for the NBAS items.

The impact of maternal anxiety disorder on mother-infant interaction in the postpartum period.

BACKGROUND: This study investigated whether postpartum anxiety disorder is associated to altered patterns of infant as well as maternal engagement in a Face-to-Face-Still-Face interaction (FFSF). SAMPLING AND METHODS: n = 39 women with postpartum DSM-IV anxiety disorder and n = 48 healthy mothers were videotaped during a FFSF with their infant (M = 4.1 months). RESULTS: Infants of the clinical group showed significantly less positive engagement during the play episode than infants of controls. This result depended on infant sex: male controls demonstrated more positive interaction than males of anxious mothers. There was no such effect for female infants who engaged significantly less positively during the play episode than males and did not change their positive engagement during the FFSF. These findings imply pronounced interactive positivity and early vulnerability to maternal anxiety symptoms in male infants. Only the infants of the controls showed the still-face effect. They also protested significantly more during the still-face, while the clinical infants' protest increased significantly during the reunion. Women of both groups did not differ in their interaction. Maternal intrusiveness was associated to infant protest in the course of the FFSF. CONCLUSIONS: Results suggest that mother-infant intervention should consider affect regulation and infant sex-specific characteristics in anxious mother-infant dyads.

Hair cortisol and lifetime discrimination: Moderation by subjective social status.

Discrimination has been associated with elevated cortisol as measured in saliva, blood, and urine. This study investigated the association between lifetime discrimination and hair cortisol concentrations, considered a measure of chronic stress. We recruited 180 young adults from diverse backgrounds. Participant responses to lifetime discrimination, home stress, and subjective status measures were recorded. Lifetime discrimination significantly predicted hair cortisol concentrations, supporting past research that discrimination experiences impact neuroendocrine systems. To our knowledge, these are the first findings associating hair cortisol concentrations with discrimination and supports prior evidence positing discrimination as a chronic stressor that serves as a risk factor for chronic disease.

The Contributions of Maternal Sensitivity and Maternal Depressive Symptoms to Epigenetic Processes and Neuroendocrine Functioning.

This study tested whether maternal responsiveness may buffer the child to the effects of maternal depressive symptoms on DNA methylation of NR3C1, 11ß-HSD2, and neuroendocrine functioning. DNA was derived from buccal epithelial cells and prestress cortisol was obtained from the saliva of 128 infants. Mothers with depressive symptoms who were more responsive and who engaged in more appropriate touch during face-to-face play had infants with less DNA methylation of NR3C1 and 11ß-HSD2 compared to mothers with depressive symptoms who were also insensitive. The combination of exposure to maternal depressive symptoms and maternal sensitivity was related to the highest prestress cortisol levels, whereas exposure to maternal depressive symptoms and maternal insensitivity was related to the lowest prestress cortisol levels.

Prenatal predictors of infant self-regulation: the contributions of placental DNA methylation of NR3C1 and neuroendocrine activity.

We examined whether placental DNA methylation of the glucocorticoid receptor gene, NR3C1 was associated with self-regulation and neuroendocrine responses to a social stressor in infancy. Placenta samples were obtained at birth and mothers and their infants (n = 128) participated in the still-face paradigm when infants were 5 months old. Infant self-regulation following the still-face episode was coded and pre-stress cortisol and cortisol reactivity was assessed in response to the still-face paradigm. A factor analysis of NR3C1 CpG sites revealed two factors: one for CpG sites 1-4 and the other for sites 5-13. DNA methylation of the factor comprising NR3C1 CpG sites 5-13 was related to greater cortisol reactivity and infant self-regulation, but cortisol reactivity was not associated with infant self-regulation. The results reveal that prenatal epigenetic processes may explain part of the development of infant self-regulation.

Assessment and evaluation of the high risk neonate: the NICU Network Neurobehavioral Scale.

There has been a long-standing interest in the assessment of the neurobehavioral integrity of the newborn infant. The NICU Network Neurobehavioral Scale (NNNS) was developed as an assessment for the at-risk infant. These are infants who are at increased risk for poor developmental outcome because of insults during prenatal development, such as substance exposure or prematurity or factors such as poverty, poor nutrition or lack of prenatal care that can have adverse effects on the intrauterine environment and affect the developing fetus. The NNNS assesses the full range of infant neurobehavioral performance including neurological integrity, behavioral functioning, and signs of stress/abstinence. The NNNS is a noninvasive neonatal assessment tool with demonstrated validity as a predictor, not only of medical outcomes such as cerebral palsy diagnosis, neurological abnormalities, and diseases with risks to the brain, but also of developmental outcomes such as mental and motor functioning, behavior problems, school readiness, and IQ. The NNNS can identify infants at high risk for abnormal developmental outcome and is an important clinical tool that enables medical researchers and health practitioners to identify these infants and develop intervention programs to optimize the development of these infants as early as possible. The video shows the NNNS procedures, shows examples of normal and abnormal performance and the various clinical populations in which the exam can be used.

Dyadic flexibility during the face-to-face still-face paradigm: a dynamic systems analysis of its temporal organization.

A dynamic systems analytical model was used to characterize infant-caregiver regulatory dynamics. Though stable, there was an increase in dyadic flexibility following a perturbation. Dyadic flexibility was positively related to infant negativity during the perturbation. Findings were qualified by infant sex and maternal depressive symptoms.

Healthy newborns' neurobehavior: norms and relations to medical and demographic factors.

OBJECTIVE: To generate neurobehavioral norms for an unselected random sample of clinically healthy newborns by examining the newborns with use of the Neonatal Intensive Care Unit Network Neurobehavioral Scale (NNNS). STUDY DESIGN: We recruited 344 healthy mothers and newborns from a well-child nursery. The NNNS, a 128-item assessment of infant neurobehavior, was used to examine newborn performance. Associations between 11 NNNS summary scales and the stress/abstinence scale, as well as medical and demographic variables, were evaluated. Mean, SD, and 5th and 95th percentile values for the summary scores of the NNNS are presented. RESULTS: NNNS scores from the 10th to the 90th percentile represent a range of normative performance. Performance on different neurobehavioral domains was related to marital status, ethnicity, prenatal, intrapartum and neonatal risk factors, complications during labor/delivery, cesarean delivery, gestational age, the age of the newborn at testing, and infant sex. CONCLUSION: These data provide clinicians and researchers with normative data for evaluation of newborn neurobehavior. Even in a low-risk sample, medical and demographic factors below clinical cut-offs were related to newborn performance. Infants with scores outside the ranges for the 11 NNNS summary scores and the stress/abstinence scale may need further observation and, if necessary, early intervention.

Behavioral epigenetics.

Sponsored by the New York Academy of Sciences, the Warren Alpert Medical School of Brown University and the University of Massachusetts Boston, "Behavioral Epigenetics" was held on October 29-30, 2010 at the University of Massachusetts Boston Campus Center, Boston, Massachusetts. This meeting featured speakers and panel discussions exploring the emerging field of behavioral epigenetics, from basic biochemical and cellular mechanisms to the epigenetic modulation of normative development, developmental disorders, and psychopathology. This report provides an overview of the research presented by leading scientists and lively discussion about the future of investigation at the behavioral epigenetic level.