Longitudinal course of endocannabinoids and N-acylethanolamines in hair of mothers and their children in the first year postpartum: investigating the relevance of maternal childhood maltreatment experiences.

BACKGROUND: Childhood maltreatment (CM) exerts long-lasting psychological and biological alterations in affected individuals and might also affect the endocannabinoid (eCB) system which modulates inflammation and the endocrine stress response. Here, we investigated the eCB system of women with and without CM and their infants using hair samples representing eCB levels accumulated during the last trimester of pregnancy and 10-12 months postpartum. METHODS: CM exposure was assessed with the Childhood Trauma Questionnaire. At both timepoints, 3 cm hair strands were collected from mothers and children (N = 170 resp. 150) to measure anandamide (AEA), 2/1-arachidonoylglycerol (2-AG/1-AG), stearoylethanolamide (SEA), oleoylethanolamide (OEA), and palmitoylethanolamide (PEA). RESULTS: Maternal hair levels of 2-AG/1-AG increased and SEA levels decreased from late pregnancy to one year postpartum. Maternal CM was associated with lower SEA levels in late pregnancy, but not one year later. In the children's hair, levels of 2-AG/1-AG increased while levels of SEA, OEA, and PEA decreased from late pregnancy to one year later. Maternal CM was not consistently associated with the eCB levels measured in children's hair. CONCLUSIONS: We provide first evidence for longitudinal change in the eCB system of mothers and infants from pregnancy to one year later. While maternal CM influenced the maternal eCB system, we found no consistent intergenerational effects on early regulation of the eCB system in children. Longitudinal research on the importance of the eCB system for the course and immunoregulation of pregnancy as well as for the children's development.

Reactivity of the Oxytocinergic and Neuroendocrine System Following the Adult Attachment Projective Picture System in Men of Recent Fatherhood: Results from an Exploratory Pilot Study with a Cross-Sectional Design.

The attachment representation (AR) of individuals affects emotional and cognitive information processes and is considered an important modulating factor of neuroendocrine stress responses. The neuropeptide oxytocin is studied as one biomolecular component underpinning this modulation. A validated procedure used in attachment-related research is the Adult Attachment Projective Picture System (AAP). To date, only a limited number of studies investigated oxytocin and neuroendocrine reactivity in the context of an attachment-related stimulus similar to the APP. In this pilot study, N = 26 men of recent fatherhood were exposed to the AAP to classify AR and to investigate salivary changes in oxytocin, cortisol and dehydroepiandrosterone (DHEA) after AAP stimulation. We observed increased oxytocin levels in response to AAP exposure, and this increase was more pronounced in fathers with unresolved/disorganized AR. No significant changes in cortisol and DHEA concentrations were observed in response to AAP administration. Interestingly, differences in basal cortisol levels (before the AAP) also depended on AR classification. Here, the group of men with unresolved/disorganized AR showed higher levels of cortisol compared to fathers with organized AR. To conclude, the finding of increased salivary oxytocin levels in response to the AAP further indicates its validity as an instrument to stimulate the attachment system.

Long-Term Consequences of Childhood Maltreatment Among Postpartum Women-Prevalence of Psychosocial Risk Factors for Child Welfare: An Independent Replication Study.

Introduction: As an especially burdensome experience, childhood maltreatment (CM) can have lifelong consequences on the mental health and wellbeing of an individual well into adulthood. We have previously reported that CM constitutes a central risk factor not only for the development of mental problems, but also for facing additional psychosocial risks, endangering healthy development of mother and offspring throughout life (e.g., financial problems, intimate partner violence, substance use). This study was designed to replicate these findings in a larger, independent study cohort. Method: In this cross-sectional replication study an independent cohort of 533 healthy postpartum women was interviewed within seven days after parturition. CM experiences were assessed retrospectively using the German version of the Childhood Trauma Questionnaire (CTQ) and current psychosocial risk factors for child welfare were assessed using the Konstanzer Index (KINDEX). Results: Of all women, 16.1% experienced emotional and 10.1% physical abuse, 28.5% emotional neglect, 9.4% physical neglect and 10.3% experienced sexual abuse. Most importantly, the higher the CM load the more psychosocial stressors existed in women's life. In Particular, women with higher CM load had a higher risk for mental health problems, intimate partner violence, financial problems, and a higher postnatal stress load. Conclusions: In an independent sample, this study replicated the previous findings that CM and psychosocial risk factors for child welfare were strongly associated in a dose-response manner. Our results emphasize the higher vulnerability of women with a CM history in the postpartum period. To avoid negative consequences for mother and child, a regular and evidence-based screening for CM and psychosocial risk factors during pregnancy and puerperium is needed to identify at-risk mothers early during pregnancy and to provide appropriate support. Hence, our findings highlight the mandatory requirement for an interdisciplinary collaboration of gynecological practices, hospitals and midwifes, along with psychologists and psychotherapists and child and youth welfare services.

Characterization of the effects of age and childhood maltreatment on ELOVL2 DNA methylation.

DNA methylation of the elongation of very long chain fatty acids protein 2 (ELOVL2) was suggested as a biomarker of biological aging, while childhood maltreatment (CM) has been associated with accelerated biological aging. We investigated the association of age and CM experiences with ELOVL2 methylation in peripheral blood mononuclear cells (PBMC). Furthermore, we investigated ELOVL2 methylation in the umbilical cord blood mononuclear cells (UBMC) of newborns of mothers with and without CM. PBMC and UBMC were isolated from 113 mother-newborn dyads and genomic DNA was extracted. Mothers with and without CM experiences were recruited directly postpartum. Mass array spectrometry and pyrosequencing were used for methylation analyses of ELOVL2 intron 1, and exon 1 and 5' end, respectively. ELOVL2 5' end and intron 1 methylation increased with higher age but were not associated with CM experiences. On the contrary, overall ELOVL2 exon 1 methylation increased with higher CM, but these changes were minimal and did not increase with age. Maternal CM experiences and neonatal methylation of ELOVL2 intron 1 or exon 1 were not significantly correlated. Our study suggests region-specific effects of chronological age and experienced CM on ELOVL2 methylation and shows that the epigenetic biomarker for age within the ELOVL2 gene does not show accelerated biological aging years after CM exposure.

Atypical maternal interaction is associated with elevated levels of hair cortisol in children.

The quality of maternal caregiving is an important factor in the healthy development of a child. One consequence of prolonged insensitive and atypical maternal interaction behavior (e.g., withdrawing from interactions with the child and role-reversal, i.e., the takeover of the parental role or parts of it by the child) in mother-child-dyads can cause alteration of the child's stress response system. Higher salivary cortisol concentrations were reported in infants and toddlers directly after negative interactions with their parents. However, no study to date has examined the association between atypical maternal interaction behavior and hair cortisol concentrations (HCC) in infants. Here, we studied the association of maternal interaction behavior with HCC of the child. Mother-child dyads (N = 112) participated in the longitudinal study My Childhood-Your Childhood. The AMBIANCE scale and its subscales were used to assess atypical maternal interaction behavior during the Strange Situation Procedure. Chronic stress levels in the child were assessed by HCC of 3 cm hair strands at the age of 12 months. Maternal educational level (operationalized in highest education level) served as a control variable. Robust multiple linear regression analyses revealed that role/boundary confusion was associated with HCC, i.e., the higher atypical interaction behavior of the mother the higher the HCC in the children. By measuring hair cortisol in this study, it is possible to determine the average long-term activity of the child's stress response system.Thus, atypical maternal interaction behavior could be a risk factor for persistent stress in children, contributing to a higher risk for negative health outcomes in later life. The results of this study highlight the importance of early intervention programs that focus on the relationship between mother and child.

Associations between childhood maltreatment and DNA methylation of the oxytocin receptor gene in immune cells of mother-newborn dyads.

The neuropeptide oxytocin (OXT) and its receptor (OXTR) modulate interpersonal relationships, particularly mother-child interactions. DNA methylation (DNAm) changes of the OXTR gene were observed in individuals who experienced Childhood Maltreatment (CM). A modulatory role of single nucleotide polymorphisms (SNP) within OXTR in association with CM on the regulation of OXTR was also postulated. Whether these CM-induced epigenetic alterations are biologically inherited by the offspring remains unknown. We thus investigated possible intergenerational effects of maternal CM exposure on DNAm and OXTR gene expression, additionally accounting for the possible influence of three SNP: rs53576 and rs2254298 (OXTR gene), and rs2740210 (OXT gene). We used the Childhood Trauma Questionnaire to classify mothers into individuals with (CM+) or without CM (CM-). Maternal peripheral immune cells were isolated from venous blood (N = 117) and fetal immune cells from the umbilical cord (N = 113) after parturition. DNA methylation was assessed using MassARRAY. Taqman assays were performed for genotyping and gene expression analyses. Among mothers, CM was not associated with OXTR mean methylation or gene expression. However, four CpG sites showed different methylation levels in CM- compared to CM+. In mothers, the OXTR rs53576 and OXT rs2740210 allelic variations interacted with CM load on the OXTR mean methylation. Maternal and newborns' mean methylation of OXTR were positively associated within CM- dyads, but not in CM+ dyads. We show gene×environment interactions on the epigenetic regulation of the oxytocinergic signaling and show the intergenerational comparability of the OXTR DNAm might be altered in infants of CM+ mothers.

Hair-based biomarkers in women with major depressive disorder: Glucocorticoids, endocannabinoids, N-acylethanolamines, and testosterone.

Background: Stress-related alterations in the regulation of several endocrine systems, including the hypothalamus-pituitary-adrenal (HPA) and -gonadal (HPG) axes and the endocannabinoid system are proposed to be involved in the etiology of major depressive disorder (MDD). Therefore, this study examines whether altered concentrations of glucocorticoids, testosterone, endocannabinoids, and related N-acylethanolamines accumulated in hair are present in MDD. Methods: Female participants (range: 19-59, Mdn = 30.5 years) were recruited, including n = 21 with a current MDD episode and n = 27 nondepressed controls without any current mental disorder. Weight-standardized samples of 3 cm hair segments were analyzed which equals to three months of retrospectively assessed biomarkers in hair. Concentrations of cortisol, cortisone, testosterone, endocannabinoids (i.e., anandamide [AEA], 2-arachidonylglycerol [2-AG]), and selected N-acylethanolamines (i.e., stearoylethanolamide [SEA], oleoylethanolamide [OEA], palmitoylethanolamide [PEA]) were measured using mass spectrometry. Results: Female MDD patients exhibited lower cortisol and cortisone levels in hair than nondepressed controls, whereas the hair concentrations of endocannabinoids, N-acylethanolamines, and testosterone did not differ between the groups. Conclusions: Our results add to the heterogeneous body of findings on alterations in hair-stored glucocorticoids and endocannabinoids in MDD. As in previous studies, there was no evidence of altered testosterone concentrations in the hair of MDD patients. Larger and longitudinal studies are needed to comprehensively explore the overall picture of endocrine alterations in MDD.

Mitochondrial bioenergetics in leukocytes and oxidative stress in blood serum of mild to moderately depressed women.

Major depressive disorder (MDD) has been associated with lower mitochondrial energy production and higher oxidative stress. We investigated whether these alterations manifest in patients with current mild to moderate MDD severity. We observed no differences in mitochondrial respiration and density (i.e., citrate-synthase activity) in peripheral blood mononuclear cells and oxidative stress markers (i.e., 8-hydroxy-2'-deoxyguanosine, 8-isoprostane) in blood serum of 20 female MDD patients compared to 24 non-depressed women. Alterations in mitochondrial energy production and oxidative stress did not linearly depend on the current severity of MDD. However, biological alterations might rather manifest with higher MDD severity/chronicity and at higher age.

Childhood maltreatment is associated with changes in mitochondrial bioenergetics in maternal, but not in neonatal immune cells.

Childhood maltreatment (CM) comprises experiences of abuse and neglect during childhood. CM causes psychological as well as biological alterations in affected individuals. In humans, it is hardly explored whether these CM consequences can be transmitted directly on a biological level to the next generation. Here, we investigated the associations between maternal CM and mitochondrial bioenergetics (mitochondrial respiration and intracellular mitochondrial density) in immune cells of mothers and compared them with those of their newborns. In n = 102 healthy mother-newborn dyads, maternal peripheral blood mononuclear cells and neonatal umbilical cord blood mononuclear cells were collected and cryopreserved shortly after parturition to measure mitochondrial respiration and intracellular mitochondrial density with high-resolution respirometry and spectrophotometric analyses, respectively. Maternal CM was assessed with the Childhood Trauma Questionnaire Maternal and neonatal mitochondrial bioenergetics were quantitatively comparable and positively correlated. Female newborns showed higher mitochondrial respiration compared to male newborns. Maternal CM load was significantly and positively associated with mitochondrial respiration and density in mothers, but not with mitochondrial respiration in newborns. Although maternal and neonatal mitochondrial bioenergetics were positively correlated, maternal CM only had a small effect on mitochondrial density in newborns, which was not significant in this study after adjustment for multiple comparisons. The biological relevance of our finding and its consequences for child development need further investigation in future larger studies. This study reports data on mitochondrial bioenergetics of healthy mother-newborn dyads with varying degrees of CM.

Yersinia enterocolitica infection of mice reveals clonal invasion and abscess formation.

Yersinia enterocolitica is a common cause of food-borne gastrointestinal disease leading to self-limiting diarrhea and mesenteric lymphadenitis. Occasionally, focal abscess formation in the livers and spleens of certain predisposed patients (those with iron overload states such as hemochromatosis) is observed. In the mouse oral infection model, yersiniae produce a similar disease involving the replication of yersiniae in the small intestine, the invasion of Peyer's patches, and dissemination to the liver and spleen. In these tissues and organs, yersiniae are known to replicate predominantly extracellularly and to form microcolonies. By infecting mice orally with a mixture of equal amounts of green- and red-fluorescing yersiniae (yersiniae expressing green or red fluorescent protein), we were able to show for the first time that yersiniae produce exclusively monoclonal microcolonies in Peyer's patches, the liver, and the spleen, indicating that a single bacterium is sufficient to induce microcolony and microabscess formation in vivo. Furthermore, we present evidence for the clonal invasion of Peyer's patches from the small intestine. The finding that only very few yersiniae are required to establish microcolonies in Peyer's patches is due to both Yersinia-specific and host-specific factors. We demonstrate that yersiniae growing in the small intestinal lumen show strongly reduced levels of invasin, the most important factor for the early invasion of Peyer's patches. Furthermore, we show that the host severely restricts sequential microcolony formation in previously infected Peyer's patches.

Detection of N-(3-oxohexanoyl)-L-homoserine lactone in mice infected with Yersinia enterocolitica serotype O8.

Yersinia enterocolitica synthesizes N-acyl-L-homoserine lactone (AHL) signal molecules via the LuxR-LuxI homologues YenR-YenI. In this study we checked two prototypes of mouse-virulent Y. enterocolitica serotype O8 strains WA-314 and 8081 for AHL production in vitro and in vivo (mouse infection model). We used thin-layer chromatography in combination with the Escherichia coli AHL biosensor to identify the AHL species produced. We detected only OHHL [N-(3-oxohexanoyl)-L-homoserine lactone] and not HHL (N-hexanoyl-L-homoserine lactone) produced by Y. enterocolitica O8 in culture supernatant or infected mouse tissue. This is the first report demonstrating AHL production by yersiniae during infection.