Increased oxidative modifications of amniotic fluid albumin in pregnancies associated with gestational diabetes mellitus.

Gestational diabetes mellitus (GDM) is a state of hyperglycaemia and increased oxidative stress with onset during pregnancy. Human serum albumin (HSA) was extracted from 26 GDM and 26 nonGDM amniotic fluid samples collected at 15 weeks gestation and analyzed by mass spectrometry. The majority of all albumin isoforms were oxidized with the cysteinylated HSA as the base peak in the deconvoluted spectrum. The HSA peak areas, from a control sample, had 36% relative standard deviation (RSD) across the six experimental days but using the relative isoform distribution improved the precision to 3-6%. The results show that the relative contribution of permanently oxidized HSA was greater (P = 0.002) and reversibly oxidized HSA was lower (P = 0.006) for GDM compared to nonGDM in the samples measured. This implies that the path toward GDM has been set prior to 15 weeks gestation and results in increased protein oxidation.

Retinal lipid and glucose metabolism dictates angiogenesis through the lipid sensor Ffar1.

Tissues with high metabolic rates often use lipids, as well as glucose, for energy, conferring a survival advantage during feast and famine. Current dogma suggests that high-energy-consuming photoreceptors depend on glucose. Here we show that the retina also uses fatty acid ß-oxidation for energy. Moreover, we identify a lipid sensor, free fatty acid receptor 1 (Ffar1), that curbs glucose uptake when fatty acids are available. Very-low-density lipoprotein receptor (Vldlr), which is present in photoreceptors and is expressed in other tissues with a high metabolic rate, facilitates the uptake of triglyceride-derived fatty acid. In the retinas of Vldlr(-/-) mice with low fatty acid uptake but high circulating lipid levels, we found that Ffar1 suppresses expression of the glucose transporter Glut1. Impaired glucose entry into photoreceptors results in a dual (lipid and glucose) fuel shortage and a reduction in the levels of the Krebs cycle intermediate α-ketoglutarate (α-KG). Low α-KG levels promotes stabilization of hypoxia-induced factor 1a (Hif1a) and secretion of vascular endothelial growth factor A (Vegfa) by starved Vldlr(-/-) photoreceptors, leading to neovascularization. The aberrant vessels in the Vldlr(-/-) retinas, which invade normally avascular photoreceptors, are reminiscent of the vascular defects in retinal angiomatous proliferation, a subset of neovascular age-related macular degeneration (AMD), which is associated with high vitreous VEGFA levels in humans. Dysregulated lipid and glucose photoreceptor energy metabolism may therefore be a driving force in macular telangiectasia, neovascular AMD and other retinal diseases.

Development of a new multi-residue laser diode thermal desorption atmospheric pressure chemical ionization tandem mass spectrometry method for the detection and quantification of pesticides and pharmaceuticals in wastewater samples.

A new solid phase extraction (SPE) method coupled to a high throughput sample analysis technique was developed for the simultaneous determination of nine selected emerging contaminants in wastewater (atrazine, desethylatrazine, 17ß-estradiol, ethynylestradiol, norethindrone, caffeine, carbamazepine, diclofenac and sulfamethoxazole). We specifically included pharmaceutical compounds from multiple therapeutic classes, as well as pesticides. Sample pre-concentration and clean-up was performed using a mixed-mode SPE cartridge (Strata ABW) having both cation and anion exchange properties, followed by analysis by laser diode thermal desorption atmospheric pressure chemical ionization coupled to tandem mass spectrometry (LDTD-APCI-MS/MS). The LDTD interface is a new high-throughput sample introduction method, which reduces total analysis time to less than 15s per sample as compared to minutes with traditional liquid-chromatography coupled to tandem mass spectrometry (LC-MS/MS). Several SPE parameters were evaluated in order to optimize recovery efficiencies when extracting analytes from wastewater, such as the nature of the stationary phase, the loading flow rate, the extraction pH, the volume and composition of the washing solution and the initial sample volume. The method was successfully applied to real wastewater samples from the primary sedimentation tank of a municipal wastewater treatment plant. Recoveries of target compounds from wastewater ranged from 78% to 106%, the limit of detection ranged from 30 to 122ng L(-1) while the limit of quantification ranged from 90 to 370ng L(-1). Calibration curves in the wastewater matrix showed good linearity (R(2)≥0.991) for all target analytes and the intraday and interday coefficient of variation was below 15%, reflecting a good precision.

Prediction of gestational diabetes mellitus based on an analysis of amniotic fluid by capillary electrophoresis.

AIM: To detect gestational diabetes mellitus biomarkers in human amniotic fluid collected for age-related genetic testing using capillary electrophoresis and a sophisticated data analysis methodology. MATERIALS & METHODS: Amniotic fluid samples were separated by capillary electrophoresis. Samples were classified using a genetic algorithm with Bayesian benefit function. The best model maximized the sensitivity and specificity and employed a leave-one-out cross-validation strategy. RESULTS: Gestational diabetes mellitus (GDM; n = 14) was distinguished from non-GDM (n = 95) with 86% sensitivity and 99% specificity using two wavelets. These wavelets were located in the unresolved protein region and on the edge of the maternally derived albumin peak. CONCLUSION: GDM is a maternal pathology; however, it was shown that it alters the biochemical profile of amniotic fluid. Testing for GDM is normally carried out at 24-28 weeks, but changes can be detected at 15 weeks gestation, suggesting that GDM onset occurs early in gestation.

Early prediction of macrosomia based on an analysis of second trimester amniotic fluid by capillary electrophoresis.

AIM: To identify, using capillary electrophoresis and chemometrics, early biomarkers in human amniotic fluid of large-for-gestational-age (LGA) infants. MATERIALS & METHODS: Second trimester amniotic fluid samples, obtained from mothers undergoing age-related amniocentesis, were analyzed by capillary electrophoresis. Electropherogram data were aligned using correlation-optimized warping. A genetic algorithm using a Bayesian evaluation function and a leave-one-out cross-validation strategy for two birth outcomes: appropriate-for-gestational-age (AGA) versus LGA infants. RESULTS: LGA (n = 23) was differentiated from AGA (n = 86) with a sensitivity of 100% and a specificity of 98% using only two wavelets. The first wavelet is associated with albumin and the second wavelet with an unknown small molecule. CONCLUSION: The approach developed herein allows LGA fetuses to be metabolically distinguished from AGA fetuses early in pregnancy and indicates that the birth of a LGA infant is already associated with an altered biochemical profile by the second trimester.