The Feasibility of the Adverse Childhood Experiences Questionnaire among Women in Danish Antenatal Care: A Mixed-Methods Study.

A traumatic upbringing increases the risks of antenatal health problems, unfavourable pregnancy outcomes, and mental disorders. Such childhood experiences may affect women's pa-renting skills and the social-emotional functioning of their children. Research on screening for adverse childhood experiences in antenatal care is limited. The objective of this study was to explore pregnant women's attitudes towards and experiences of an adverse childhood experiences questionnaire, and to assess the relevance of the questionnaire among a population of pregnant women referred to antenatal care levels one and two, targeting women who are generally not perceived to be vulnerable. Data were collected at three maternity wards and consisted of quantitative data on 1352 women's adverse childhood experience scores, structured observations of 18 midwifery visits, and in-depth interviews with 15 pregnant women. Quantitative data were analysed by descriptive statistics, and qualitative data were analysed using systematic text condensation. The qualitative analysis revealed two main categories: "Being screened for childhood adversities" and "Having adverse childhood experiences". In the study population, the prevalence of adverse childhood experiences was high. The women assessed the adverse childhood experiences questionnaire to be a relevant and acceptable screening method. Furthermore, women's perceptions of their relationship with their midwife greatly impacted their attitudes towards and experiences of the questionnaire.

Fractures in Childhood and Young Adulthood According to Maternal Smoking in Late Pregnancy. A Danish Cohort Study.

BACKGROUND: Fractures account for the most frequent cause of hospitalization during childhood and numbers have increased over time. Of all fractures in childhood and young adulthood, 66% are recurrent fractures, suggesting that some people are predestined for fractures. The aim of this study was to investigate the association between maternal smoking during late pregnancy and the risk of fractures in the children. METHODS: The study included 11,082 mothers and their children from the cohort "Healthy Habits for Two" born between 1984 and 1987. Information about maternal smoking during pregnancy came from questionnaires filled out in pregnancy, while information about fractures was derived from the Danish National Patient Registry. Over a follow-up of 24 years (1994-2018), Cox regression with multiple failures was used to estimate hazard ratios (HRs) with 95% confidence intervals (CIs) for fractures in childhood and young adulthood according to maternal smoking in late pregnancy. Information about body mass index (BMI) and smoking status in young adulthood was included as time variant covariates. RESULTS: During an age span of 8-32 years, 6,420 fractures were observed. Of the mothers, 39.1% smoked during late pregnancy. Compared to children of mothers who did not smoke, children of mothers who smoked 1-9 cigarettes per day and 10+ cigarettes per day had an increased risk of fractures (HR 1.14 [CI: 1.06; 1.21] and HR 1.14 [CI: 1.07; 1.22], respectively). After adjusting for BMI and smoking status in young adulthood, the findings were slightly strengthened, showing an increased risk of fractures of 23 and 25% in children of mothers smoking 1-9 cigarettes per day and 10+ cigarettes per day, respectively. CONCLUSION: Maternal smoking during late pregnancy was associated with a higher risk of fractures in the child. This result indicates that exposure to cigarette smoke in utero may play a role in lifelong bone health.

Identification of 47 novel mutations in patients with Alport syndrome and thin basement membrane nephropathy.

BACKGROUND: Alport syndrome (ATS) is a progressive hereditary nephropathy characterized by hematuria and proteinuria. It can be associated with extrarenal manifestations. In contrast, thin basement membrane nephropathy (TBMN) is characterized by microscopic hematuria, is largely asymptomatic, and is rarely associated with proteinuria and end-stage renal disease. Mutations have been identified in the COL4A5 gene in ATS and in the COL4A3 and COL4A4 genes in ATS and TBMN. To date, more than 1000 different mutations in COL4A5, COL4A3, and COL4A4 are known. METHODS: In this study mutational analysis by exon sequencing and multiplex ligation-dependent probe amplification was performed in a large European cohort of families with ATS and TBMN. RESULTS: Molecular diagnostic testing of 216 individuals led to the detection of 47 novel mutations, thereby expanding the spectrum of known mutations causing ATS and TBMN by up to 10 and 6%, respectively, depending on the database. Remarkably, a high number of ATS patients with only single mutations in COL4A3 and COL4A4 were identified. Additionally, three ATS patients presented with synonymous sequence variants that possible affect correct mRNA splicing, as suggested by in silico analysis. CONCLUSIONS: The results of this study clearly broaden the genotypic spectrum of known mutations for ATS and TBMN, which will in turn now facilitate future studies into genotype-phenotype correlations. Further studies should also examine the significance of single heterozygous mutations in COL4A3 and COL4A4 and of synonymous sequence variants associated with ATS.

Dealing with the incidental finding of secondary variants by the example of SRNS patients undergoing targeted next-generation sequencing.

BACKGROUND: Steroid-resistant nephrotic syndrome (SRNS) is a severe cause of progressive renal disease. Genetic forms of SRNS can present with autosomal recessive or autosomal dominant inheritance. Recent studies have identified mutations in multiple podocyte genes responsible for SRNS. Improved sequencing methods (next-generation sequencing, NGS) now promise rapid mutational testing of SRNS genes. METHODS: In the present study, a simultaneous screening of ten SRNS genes in 37 SRNS patients was performed by NGS. RESULTS: In 38 % of the patients, causative mutations in one SRNS gene were found. In 22 % of the patients, in addition to these mutations, a secondary variant in a different gene was identified. CONCLUSIONS: This high incidence of accumulating sequence variants was unexpected but, although they might have modifier effects, the pathogenic potential of these additional sequence variants seems unclear so far. The example of molecular diagnostics by NGS in SRNS patients shows that these new sequencing technologies might provide further insight into molecular pathogenicity in genetic disorders but will also generate results, which will be difficult to interpret and complicate genetic counseling. Although NGS promises more frequent identification of disease-causing mutations, the identification of causative mutations, the interpretation of incidental findings and possible pitfalls might pose problems, which hopefully will decrease by further experience and elucidation of molecular interactions.

Next generation sequencing as a useful tool in the diagnostics of mosaicism in Alport syndrome.

Alport syndrome (ATS) is a progressive hereditary nephropathy characterized by hematuria and/or proteinuria with structural defects of the glomerular basement membrane. It can be associated with extrarenal manifestations (high-tone sensorineural hearing loss and ocular abnormalities). Somatic mutations in COL4A5 (X-linked), COL4A3 and COL4A4 genes (both autosomal recessive and autosomal dominant) cause Alport syndrome. Somatic mosaicism in Alport patients is very rare. The reason for this may be due to the difficulty of detection. We report the case of a boy and his mother who presented with Alport syndrome. Mutational analysis showed the novel hemizygote pathogenic mutation c.2396-1G>A (IVS29-1G>A) at the splice acceptor site of the intron 29 exon 30 boundary of the COL4A5 gene in the boy. The mutation in the mother would not have been detected by Sanger sequencing without the knowledge of the mutational analysis result of her son. Further investigation of the mother using next generation sequencing showed somatic mosaicism and implied potential germ cell mosaicism. The mutation in the mother has most likely occurred during early embryogenesis. Analysis of tissue of different embryonic origin in the mother confirmed mosaicism in both mesoderm and ectoderm. Low grade mosaicism is very difficult to detect by Sanger sequencing. Next generation sequencing is increasingly used in the diagnostics and might improve the detection of mosaicism. In the case of definite clinical symptoms of ATS and missing detection of a mutation by Sanger sequencing, mutational analysis should be performed by next generation sequencing.