Incidental nutrient transfers: Assessing critical times in agricultural catchments using high-resolution data

AbstractManaging incidental losses associated with liquid slurry applications during closed periods has significant cost and policy implications and the environmental data required to review such a measure are difficult to capture due to storm dependencies. Over four years (2010–2014) in five intensive agricultural catchments, this study used high-resolution total and total reactive phosphorus (TP and TRP), total oxidised nitrogen (TON) and suspended sediment (SS) concentrations with river discharge data to investigate the magnitude and timing of nutrient losses. A large dataset of storm events (defined as 90th percentile discharges), and associated flow-weighted mean (FWM) nutrient concentrations and TP/SS ratios, was used to indicate when losses were indicative of residual or incidental nutrient transfers. The beginning of the slurry closed period was reflective of incidental and residual transfers with high storm FWM P (TP and TRP) concentrations, with some catchments also showing elevated storm TP:SS ratios. This pattern diminished at the end of the closed period in all catchments. Total oxidised N behaved similarly to P during storms in the poorly drained catchments and revealed a long lag time in other catchments. Low storm FWM P concentrations and TP:SS ratios during the weeks following the closed period suggests that nutrients either weren't applied during this time (best times chosen) or that they were applied to less risky areas (best places chosen). For other periods such as late autumn and during wet summers, where storm FWM P concentrations and TP:SS ratios were high, it is recommended that an augmentation of farmer knowledge of soil drainage characteristics with local and detailed current and forecast soil moisture conditions will help to strengthen existing regulatory frameworks to avoid storm driven incidental nutrient transfers

The Dynamic Role of Sphingolipids and their Regulatory Enzymes in Preeclampsia

Sphingolipids act as regulators of cell fate decisions. Ceramides (CERs) are key effector molecules regulating cell death. CERs metabolism is controlled by a balance between its breakdown and synthesis by the enzymes acid ceramidase (AC) and acid sphingomyelinase (ASM), respectively. We hypothesized that sphingolipid metabolism plays a role in proper placental development by establishing trophoblast cell fate and alterations in the pathways contribute to preeclampsia. CERs levels were increased in preeclamptic relative to normotensive preterm control placentae and this associated with decreased AC and ASM expression levels and altered glycosylation and activity of these enzymes, an effect caused by oxidative stress. Furthermore, C-16 ceramide or inhibiting AC activity induced autophagy in placental cells. In conclusion, altered expression of AC and ASM in preeclamptic placentae, induced by oxidative stress, are responsible for changes in the sphingolipid rheostat which may contribute to increased autophagy and trophoblast cell turnover characteristic of this disorder.MAS

Ceramide-Induced Lysosomal Biogenesis and Exocytosis in Early-Onset Preeclampsia Promotes Exosomal Release of SMPD1 Causing Endothelial Dysfunction

Aberrant ceramide build-up in preeclampsia, a serious disorder of pregnancy, causes exuberant autophagy-mediated trophoblast cell death. The significance of ceramide accumulation for lysosomal biogenesis in preeclampsia is unknown. Here we report that lysosome formation is markedly increased in trophoblast cells of early-onset preeclamptic placentae, in particular in syncytiotrophoblasts. This is accompanied by augmented levels of transcription factor EB (TFEB). In vitro and in vivo experiments demonstrate that ceramide increases TFEB expression and nuclear translocation and induces lysosomal formation and exocytosis. Further, we show that TFEB directly regulates the expression of lysosomal sphingomyelin phosphodiesterase (L-SMPD1) that degrades sphingomyelin to ceramide. In early-onset preeclampsia, ceramide-induced lysosomal exocytosis carries L-SMPD1 to the apical membrane of the syncytial epithelium, resulting in ceramide accumulation in lipid rafts and release of active L-SMPD1 via ceramide-enriched exosomes into the maternal circulation. The SMPD1-containing exosomes promote endothelial activation and impair endothelial tubule formation in vitro. Both exosome-induced processes are attenuated by SMPD1 inhibitors. These findings suggest that ceramide-induced lysosomal biogenesis and exocytosis in preeclamptic placentae contributes to maternal endothelial dysfunction, characteristic of this pathology

Disruption of sphingolipid metabolism augments ceramide-induced autophagy in preeclampsia

Bioactive sphingolipids including ceramides are involved in a variety of pathophysiological processes by regulating cell death and survival. The objective of the current study was to examine ceramide metabolism in preeclampsia, a serious disorder of pregnancy characterized by oxidative stress, and increased trophoblast cell death and autophagy. Maternal circulating and placental ceramide levels quantified by tandem mass spectrometry were elevated in pregnancies complicated by preeclampsia. Placental ceramides were elevated due to greater de novo synthesis via high serine palmitoyltransferase activity and reduced lysosomal breakdown via diminished ASAH1 expression caused by TGFB3-induced E2F4 transcriptional repression. SMPD1 activity was reduced; hence, sphingomyelin degradation by SMPD1 did not contribute to elevated ceramide levels in preeclampsia. Oxidative stress triggered similar changes in ceramide levels and acid hydrolase expression in villous explants and trophoblast cells. MALDI-imaging mass spectrometry localized the ceramide increases to the trophophoblast layers and syncytial knots of placentae from pregnancies complicated by preeclampsia. ASAH1 inhibition or ceramide treatment induced autophagy in human trophoblast cells via a shift of the BOK-MCL1 rheostat toward prodeath BOK. Pharmacological inhibition of ASAH1 activity in pregnant mice resulted in increased placental ceramide content, abnormal placentation, reduced fetal growth, and increased autophagy via a similar shift in the BOK-MCL1 system. Our results reveal that oxidative stressinduced reduction of lysosomal hydrolase activities in combination with elevated de novo synthesis leads to ceramide overload, resulting in increased trophoblast cell autophagy, and typifies preeclampsia as a sphingolipid storage disorder

A Single Sphingomyelin Species Promotes Exosomal Release of Endoglin into the Maternal Circulation in Preeclampsia

Preeclampsia (PE), an hypertensive disorder of pregnancy, exhibits increased circulating levels of a short form of the auxillary TGF-beta (TGFB) receptor endoglin (sENG). Until now, its release and functionality in PE remains poorly understood. Here we show that ENG selectively interacts with sphingomyelin(SM)-18:0 which promotes its clustering with metalloproteinase 14 (MMP14) in SM-18:0 enriched lipid rafts of the apical syncytial membranes from PE placenta where ENG is cleaved by MMP14 into sENG. The SM-18:0 enriched lipid rafts also contain type 1 and 2 TGFB receptors (TGFBR1 and TGFBR2), but not soluble fms-like tyrosine kinase 1 (sFLT1), another protein secreted in excess in the circulation of women with PE. The truncated ENG is then released into the maternal circulation via SM-18:0 enriched exosomes together with TGFBR1 and 2. Such an exosomal TGFB receptor complex could be functionally active and block the vascular effects of TGFB in the circulation of PE women