BACKGROUND/OBJECTIVES:Congenital heart defects (CHDs) are associated with disruptions in one-carbon metabolism. In a family-based trio design, we investigated whether plasma concentrations of choline, betaine, and folate are associated with CHD severity. METHODS:The study included 72 children with CHD, 69 of their mothers and 64 of the fathers. CHD clinical severity was classified according to the European network of population-based registries for the epidemiological surveillance of congenital anomalies (EUROCAT) system and the German PAN study (Prevalence of Congenital Heart Defects in Newborns). Concentrations of choline, betaine, and folates were quantified in plasma and urine samples from a subgroup of the participants. RESULTS:The children [mean (SD) age 3.1 (3.2) years, 59.7% males] presented with varying CHD severities according to EUROCAT (62.5% severe and 37.5% mild) and PAN classifications (45.8% severe, 30.6% moderate and 23.6% mild). The means (SD) of plasma concentrations of choline were 14.0 (10.0) µmol/L in the children, 9.5 (5.1) µmol/L in the mothers and 10.3 (5.4) µmol/L in the fathers. Plasma choline concentrations < 10 µmol/L were observed in 38 mothers (66.7%) and were associated with having a child with severe CHD [adjusted odds ratio (aOR) 3.7; 95% confidence intervals (95%CIs) = 1.1, 12.2] compared to mothers with choline ≥ 10 µmol/L. Lowered plasma choline concentrations were detected in 27 fathers (62.8%) and were also associated with severe CHD (aOR 7.4; 95%CIs = 1.7, 31.5). Child concentrations of choline, betaine and folate and parents' concentrations of betaine and folate were not associated with disease severity. CONCLUSIONS:Lower plasma choline in the parents detectable several years after conception was related to having a child with severe CHD compared to families of children with higher plasma choline. Maternal and paternal choline metabolism may have a role in modulating CHD severity. Etiological studies aiming at the prevention of congenital anomalies should focus on maternal and paternal risk factors in the preconception and early pregnancy.

Background/Objectives: Congenital heart defects (CHD) are the most common structural birth defects that exhibit high heritability. Emerging evidence suggested that CHD are as-sociated with disruptions in one-carbon metabolism. In a family-based trio design, we in-vestigated whether maternal, paternal, and child plasma concentrations of choline, beta-ine, and folate were associated with CHD severity. Subjects and Methods: The study in-cluded 72 children with CHD, 69 mothers and 64 fathers of the children. CHD severity was classified according to the European network of population-based registries for the epidemiological surveillance of congenital anomalies (EUROCAT) system and the Ger-man PAN study (Prevalence of Congenital Heart Defects in Newborns). Plasma and urine concentrations of choline and betaine and plasma folate vitamers were quantified using ultra-performance liquid chromatography–tandem mass spectrometry. Results: The chil-dren [mean (SD) age 3.1 (3.2) years, 59.7% males] presented with varying CHD severities according to EUROCAT (62.5% severe and 37.5% mild) and PAN classifications (45.8% severe, 30.6% moderate and 23.6% mild). Plasma concentrations of choline were < 10 µmol/L in 38 (66.7%) of the mothers and 27 (62.8%) of the fathers who provided blood samples. Maternal plasma choline concentrations < 10 µmol/L were associated with hav-ing a child with severe CHD [adjusted odds ratio (aOR) 3.7; 95% confidence intervals (95%CI) = 1.1, 12.2 compared to mothers with choline concentrations ≥ 10 µmol/L]. Low-ered paternal plasma choline concentrations were also associated with severe CHD (aOR 7.4; 95% CI = 1.7, 31.5). Plasma concentrations of choline in the children and those of be-taine and folate vitamers in parents and children were not associated with CHD severity. Conclusions: Lower plasma concentrations of choline in the parents detectable several years after conception, were related to having a child with severe CHD compared with families of children with higher plasma choline. These findings support a potential role for maternal and paternal choline metabolism in modulating CHD severity. Etiological studies aiming at prevention of prevalent congenital anomalies should focus on maternal and paternal risk factors.

U87MG, U373MG and U343MG cells were embedded in a 3-dimensional matrix of fibrin and monitored over 3 days using real-time video microscopy

Expression of integrin β1 and integrin β3 affects invadopodia formation and growth of primary glioblastoma cells in fibrin clot and plasma clot

Expression of fibrin(ogen) and its clotting product fibrin in glioma tumor tissues

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FAK activation in glioblastoma cells embedded in fibrin clot

Background: Anemia and growth impairment are major health problems in children under 5 years in countries with ongoing crises. Aim : The aim of this study is to investigate the prevalence of anemia; low z scores for height for age, weight for age, and weight for height; and elevated plasma total homocysteine (tHcy) and low folate and vitamin B 12 concentrations in Syrian children aged 12–60 months. Methods : This cross‐sectional study included 344 children of whom physical growth was measured. Blood count and circulating biomarkers of folate and B 12 and blood count were measured in a subgroup of the children. Data were collected on sociodemographic and contextual factors. Results: Anemia (age‐specific cutoff values for hemoglobin according to the World Health Organization) was detected in 24.4% of the children. The height‐for‐age z score was < −2 in 32.0%, the weight‐for‐age z score was < −2 in 16.3%, and the weight‐for height z sore< −2 in 1.5% of the children. The concentrations of tHcy, folate, and vitamin B 12 were (median and (10th, 90th percentiles)) 8.4 (5.6, 12.3) μ mol/L, 24.0 (9.3, 34.7) nmol/L, and 198 (123, 367) pmol/L, respectively. Anemia and child anthropometric measures were not associated with elevated tHcy, low folate, or low B 12 concentrations. Conclusion: Anemia and low height‐ or weight‐to‐age z scores are highly prevalent among Syrian preschool children. Low folate or vitamin B 12 status was common, but they did not explain these conditions. Future studies may investigate whether early life multinutrient interventions may improve child growth and anemia.

U343MG cells on day 1 after embedding in fibrin

U87MG cells on day 1 after embedding in fibrin

Effect of thrombin and CaCl2 on glioblastoma growth in suspension or embedded in 3D matrigel.

U87MG cells on day 3 after embedding in fibrin

U373MG cells on day 1 after embedding in fibrin

Representative MR images after 2 weeks of glioma grown in mice

Clinicopathologic data of tissue donors

Invadopodia formation of GBM cells in fibrin clot and plasma clot depends on integrins β1 and β3

Expression of Integrin β1 and integrin β3 in gliomas based on IDH1 and EGFR mutation status

Background: Patients suffering from chronic kidney disease (CKD) have a high risk of premature cardiovascular morbidity and mortality. It has been suggested that elevated homocysteine (Hcy) or disturbances in the transmethylation pathway may contribute to this high cardiovascular risk burden due to epigenetic mechanisms. The objective of this study was to explore the prognostic value of Hcy, S-adenosylhomocysteine (SAH) and S-adenosylmethionine (SAM) (one-carbon (C1)-metabolites) among patients with CKD. Methods: Plasma concentrations of Hcy, SAM and SAH were measured among 297 participants with CKD (KDIGO GFR category G2-G5). The predefined endpoint was the occurrence of major cardiovascular events (MACE), defined as carotid, coronary and peripheral arterial revascularization, stroke, acute myocardial infarction, major amputation, cardiovascular death and all-cause mortality during a median (IQR) follow-up period of 4.0 [3.2; 4.3] years. Results: Among all participants, the median (IQR) of plasma Hcy, SAH, and SAM levels were 16.6 [13.5; 21.2] µmol/L, 41.5 [26.6; 63.9] nmol/L, 183.4 [151.1; 223.5] nmol/L, respectively. Estimated glomerular filtration rate (eGFR) correlated more strongly with plasma SAH (r = -0.588) than with SAM (r = -0.497) and Hcy (r = -0.424). During the follow-up period, 55 participants experienced MACE. In a univariate Kaplan Meier analysis, all three C1-metabolites were significantly associated with the occurrence of the primary outcome. In a Cox-regression analysis, the association between Hcy and MACE was not significant after adjustment for age and sex (hazard ratio (HR) and 95% confidence intervals (95% CI) for the 3rd vs. 1st tertile = 1.804 (0.868-3.974)). Both SAH and SAM were not associated with MACE after adjustment for age, sex and additionally for renal function markers (SAH: HR 3rd vs. 1st tertile 1.645 95% (0.654-4.411); SAM: HR 3rd vs. 1st tertile 1.920 95% CI (0.764-5.138)). Conclusions: In people with CKD, plasma Hcy, SAH and SAM were not independent predictors of MACE after adjustment for age, sex and renal function. Disturbed renal function may explain elevated C1-metabolites and disturbed transmethylation, while this pathway is not likely to be an appropriate access point to modify the risk of cardiovascular events in CKD patients.

U87MG cells on day 1 after embedding in matrigel

Clotting correlates with GBM progression. A and B, Fibrin formation was assessed in tumor tissues from patients with astrocytoma grade 2 (A2; n = 3), astrocytoma grade 3 (A3; n = 3), or GBM (n = 3) compared with healthy brain tissue (Control; n = 3) by fluorescence microscopy. A, Representative images of fibrinogen (green) in GBM and healthy brain tissue are shown. Nuclei are stained with DAPI (blue). Scale bar, 100 μm. B, Percentage of fibrin(ogen)-positive areas per optical field. C–E, Primary tumor cells freshly isolated from patients with astrocytoma grade 2 (A2, n = 2), astrocytoma grade 3 (A3, n = 3), or GBM (n = 12) were embedded in a 3D matrix of PC, FC, or Matrigel BM. Representative phase-contrast images are shown after 4 days in high magnification. Black arrows show invadopodia. Scale bar, 100 μm (C). D, Invadopodia-positive tumor cells were counted as percent of total per optical field after 4 days of embedding in PC, FC, or BM. E, Proliferation was assessed per optical field as fold increase of cells after 4 days of embedding. The dotted line reflects baseline cell numbers. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. BM, basement membrane; FC, fibrin clot; n.s., nonsignificant; PC, plasma clot.