Expression of <i>lcat</i>, but not <i>cetp</i>, is significantly decreased in <i>apoc2</i> mutant zebrafish.

<p>In situ hybridization (A) and qPCR (B) results showing <i>lcat</i> and <i>cetp</i> mRNA expression in 5.3 dpf zebrafish embryos. mRNA expression of <i>apoc2</i>, <i>lcat</i> and <i>cetp</i> (C) and <i>apoa1</i>, <i>apoe</i>, <i>apob</i> and <i>mtp</i> (D) in adult zebrafish liver. Results are mean±s.e.m.; numbers of biological replicates are indicated on the graphs; *P<0.05, **P<0.01 and ***P<0.001 (Student’s t-test).</p

Reducing hypertriglyceridemia does not correct the FC/CE ratio.

<p>Plasma TG levels (A), cholesterol levels (B) and the FC/CE ratio (C) in WT, <i>apoc2</i> mutants and the <i>apoc2</i> mutants fed a low fat diet (LFD). Results are mean±s.e.m.; n = 3 in each group; *P<0.05, ***P<0.001 (Student’s t-test).</p

Patients with familial chylomicronemia syndrome (FCS) have disproportional FC and CE levels in plasma.

<p>TG levels (A), cholesterol levels (B) and the FC/CE ratio (C) in plasma of healthy subjects (Ctrl), in FCS patients’ whole plasma (FCS-W), and in chylomicron-depleted FCS plasma (FCS-CD). (D) A linear regression analysis of the FC/CE ratio and plasma TG levels (n = 4 in control group and n = 5 in FCS-W and FCS-CD groups). (E) LCAT activity in healthy subjects’ and FCS-CD plasma (n = 5 in control group and n = 6 in FCS-CD group). Results are mean±s.e.m.; *P<0.05, **P<0.01 and ***P<0.001 (Student’s t-test).</p

Clustering of MetS-defining variables with visceral fat or waist circumference.

<p>Principal component analysis (PCA) was performed with either visceral fat (VF) or waist circumference (WC) and remaining four MetS-defining variables: systolic blood pressure (SBP) and fasting serum concentrations of triglycerides (TG), HDL-cholesterol (HDL-chol) and glucose (Glu). To examine whether identified principal components vary across the cardiovascular protocol, the analysis was repeated for each of its five sections (i.e., supine, standing, sitting, stress and stress recovery). Principal components with eigenvalue>1 and loadings of individual MetS-defining variables ≥0.3 were considered significant <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0082368#pone.0082368-Woolston1" target="_blank">[33]</a>. These analyses were done in males and females separately and males and females together adjusting for sex (sex-pooled analyses). All variables were adjusted for age and when relevant for height prior to sex-separate PCA and additionally for sex prior to sex-pooled PCA.</p

Basic characteristics and main outcomes in studied adolescent males and females.

<p>BMI: body-mass index. Non-adjusted means and standard deviations are presented. Sex differences were evaluated with Student T-test or with non-parametric Wilcoxon test when data were not normally distributed*.</p

Relationships of VF with SF in adolescent males and females.

<p>A) Magnetic resonance images of analyzed umbilical slices in 2 individuals with similar subcutaneous fat and different visceral fat. B) Univariate correlations between of VF with SF are shown in adolescent males and females. </p

BMI and waist circumference as predictors of VF- and SF-specific quantities (VF and SF adjusted for each other).

<p>Multivariate linear regression models examining the relationships of BMI and waist circumference with each VF and SF (while adjusting for each other) are shown in adolescent males and females. All relationships were also adjusted for potentially confounding effects of age and height when appropriate.</p

Placental lipoprotein lipase DNA methylation alterations are associated with gestational diabetes and body composition at 5 years of age

<p>Gestational diabetes mellitus (GDM) is associated with obesity in childhood. This suggests that consequences of <i>in utero</i> exposure to maternal hyperglycemia extend beyond the fetal development, possibly through epigenetic programming. The aims of this study were to assess whether placental DNA methylation (DNAm) marks were associated with maternal GDM status and to offspring body composition at 5 years old in a prospective birth cohort. DNAm levels were measured in the fetal side of the placenta in 66 samples (24 from GDM mothers) using bisDNA-pyrosequencing. Anthropometric and body composition (bioimpedance) were measured in children at 5 years of age. Mann-Whitney and Spearman tests were used to assess associations between GDM, placental DNAm levels at the <i>lipoprotein lipase</i> (<i>LPL</i>) locus and children's weight, height, body mass index (BMI), body fat, and lean masses at 5 years of age. Weight, height, and BMI z-scores were computed according to the World Health Organization growth chart. Analyses were adjusted for gestational age at birth, child sex, maternal age, and pre-pregnancy BMI. <i>LPL</i> DNAm levels were positively correlated with birth weight z-scores (r = 0.252, <i>P</i> = 0.04), and with mid-childhood weight z-scores (r = 0.314, <i>P</i> = 0.01) and fat mass (r = 0.275, <i>P</i> = 0.04), and negatively correlated with lean mass (r = −0.306, <i>P</i> = 0.02). We found a negative correlation between <i>LPL</i> DNAm and mRNA levels in placenta (r = −0.459; <i>P</i> < 0.001), which highlights the regulation of transcriptional activity by these epivariations. We demonstrated that alterations in fetal placental DNAm levels at the <i>LPL</i> gene locus are associated with the anthropometric profile in children at 5 years of age. These findings support the concept of fetal metabolic programming through epigenetic changes.</p

Additional file 1: Figure S1. of PPARGC1α gene DNA methylation variations in human placenta mediate the link between maternal hyperglycemia and leptin levels in newborns

Loci analysed in E-21. PRDM16 (A), BMP7 (B), CTBP2 (C) and PPARGC1α (D) genes CpGs epigenotyped are shown. The CpGs within BMP7-A. CTBP2-A and PPARGC1α-A locus were significantly well correlated with each other. For Gen3G, when the CpGs identified in E-21 was covered by 450k array probesets, the exact same CpGs were selected (*cg01046951; ≠cg04873098; ¥cg08550435). Since some CpGs were not covered by the 450k array, probesets covering variable CpGs in close vicinity to those identified in E-21 were selected. PRDM16: cg06814194 (1st intron) and cg23738647 (exon 6); BMP7: cg18759209 (proximal promoter); and PPARGC1α: cg11270806 and cg27514608 (both intron 5). (PDF 180 kb

Additional file 3: Table S2. of PPARGC1α gene DNA methylation variations in human placenta mediate the link between maternal hyperglycemia and leptin levels in newborns

Spearman’s correlation coefficients between placental DNA methylation and metabolic variables of newborns in E-21 and Gen3G cohorts. (PDF 268 kb