Pb(II) Induces Scramblase Activation and Ceramide-Domain Generation in Red Blood Cells

The mechanisms of Pb(II) toxicity have been studied in human red blood cells using confocal microscopy, immunolabeling, fluorescence-activated cell sorting and atomic force microscopy. The process follows a sequence of events, starting with calcium entry, followed by potassium release, morphological change, generation of ceramide, lipid flip-flop and finally cell lysis. Clotrimazole blocks potassium channels and the whole process is inhibited. Immunolabeling reveals the generation of ceramide-enriched domains linked to a cell morphological change, while the use of a neutral sphingomyelinase inhibitor greatly delays the process after the morphological change, and lipid flip-flop is significantly reduced. These facts point to three major checkpoints in the process: first the upstream exchange of calcium and potassium, then ceramide domain formation, and finally the downstream scramblase activation necessary for cell lysis. In addition, partial non-cytotoxic cholesterol depletion of red blood cells accelerates the process as the morphological change occurs faster. Cholesterol could have a role in modulating the properties of the ceramide-enriched domains. This work is relevant in the context of cell death, heavy metal toxicity and sphingolipid signaling.AGA was a predoctoral student supported by the Basque Government and later by the University of the Basque Country (UPV/EHU). This work was also supported in part by grants from the Spanish Government (FEDER/MINECO BFU 2015-66306-P to F.M.G. and A.A.) and the Basque Government (IT849-13 to F.M.G. and IT838-13 to A.A.), and by the Swiss National Science Foundation

A Mixed Methods Approach for Fuel Characterisation in Gorse (<i>Ulex europaeus</i> L.) Scrub from High-Density UAV Laser Scanning Point Clouds and Semantic Segmentation of UAV Imagery

The classification and quantification of fuel is traditionally a labour-intensive, costly and often subjective operation, especially in hazardous vegetation types, such as gorse (Ulex europaeus L.) scrub. In this study, unmanned aerial vehicle (UAV) technologies were assessed as an alternative to traditional field methodologies for fuel characterisation. UAV laser scanning (ULS) point clouds were captured, and a variety of spatial and intensity metrics were extracted from these data. These data were used as predictor variables in models describing destructively and non-destructively sampled field measurements of total above ground biomass (TAGB) and above ground available fuel (AGAF). Multiple regression of the structural predictor variables yielded correlations of R2 = 0.89 and 0.87 for destructively sampled measurements of TAGB and AGAF, respectively, with relative root mean square error (RMSE) values of 18.6% and 11.3%, respectively. The best metrics for non-destructive field-measurements yielded correlations of R2 = 0.50 and 0.49, with RMSE values of 40% and 30.8%, for predicting TAGB and AGAF, respectively, indicating that ULS-derived structural metrics offer higher levels of precision. UAV-derived versions of the field metrics (overstory height and cover) predicted TAGB and AGAF with R2 = 0.44 and 0.41, respectively, and RMSE values of 34.5% and 21.7%, demonstrating that even simple metrics from a UAV can still generate moderate correlations. In further analyses, UAV photogrammetric data were captured and automatically processed using deep learning in order to classify vegetation into different fuel categories. The results yielded overall high levels of precision, recall and F1 score (0.83 for each), with minimum and maximum levels per class of F1 = 0.70 and 0.91. In conclusion, these ULS-derived metrics can be used to precisely estimate fuel type components and fuel load at fine spatial resolutions over moderate-sized areas, which will be useful for research, wildfire risk assessment and fuel management operations

Lactoferrin and lactoferricin endocytosis halt Giardia cell growth and prevent infective cyst production

Abstract Lactoferrin (LF) is an 80 KDa iron-binding glycoprotein that plays a significant role in the innate immune system and is considered to be an important microbicide molecule. It has been suggested to be effective in the treatment of giardiasis, an intestinal disease caused by the protozoan parasite G. lamblia. However, the molecular mechanisms by which LF exerts its effect on this parasite are unknown. Most of the microbicidal activity of human or bovine LF (hLF or bLF) has been associated with the N-terminal region of the mature LF - lactoferricin (LFcin). LFcin is produced by pepsin cleavage of the native protein in vitro and likely in vivo. In this work, we analyse the participation of the endocytic machinery of G. lamblia in the internalization of bLF and bLFcin and their effects on cell homeostasis. Our results show that, when bLF or bLFcin are internalized by receptor-mediated endocytosis, cell growth stops, and morphological changes are produced in the trophozoites, which ultimately will produce immature cysts. Our findings contribute to disclose the fine mechanism by which bLF and bLFcin may function as an antigiardial molecule and why they have therapeutic potential to eradicate giardiasis