A fast and reliable method for the determination of polycyclic aromatic hydrocarbons (PAHs) in water samples by solid-phase extraction (SPE) using centrifugation has been developed and optimized. A silica-based C18 cartridge was used; parameters affecting the extraction procedure such as type and volume of the elution solvent, breakthrough volume of the percolated water sample, drying of the sorbent, and evaporation of the elute have been studied. The innovation of this work was the examination of the use of a centrifugation technique in both the drying and elution steps. When combined with centrifugation, the volume of the elution solvent was reduced to 1 mL and the time for sorbent drying decreased also to 10 min under vacuum. Under optimal conditions, recoveries for the 16 U.S. EPA PAHs were between 70 and 85% and the relative standard deviation varied between 1 and 14%. Surrogate standard recoveries were similarly between 61 and 94% with a relative standard deviation between 2 and 15%. The simplicity of the described method, use of less of organic solvent, short procedure time, and good recoveries demonstrate the advantages of this environmentally friendly approach for routine analysis of numerous samples.
Recent experimental findings suggest the involvement of the 26S proteasome, the main protease active in eukaryotic cells, in the process that leads mammalian sperm to become fully fertile, so-called capacitation. Unfortunately, its role in male gametes signaling is still far from being completely understood. For this reason, here, we realized a computational model, based on network theory, with the aim of rebuilding and exploring its signaling cascade. As a result, we found that the 26S proteasome is part of a signal transduction system that recognizes the bicarbonate ion as an input terminal and two intermediate layers of information processing. The first is under the control of the 26S proteasome and protein kinase A (PKA), which are strongly interconnected, while the latter depends on intracellular calcium concentrations. Both are active in modulating sperm function by influencing the protein phosphorylation pattern and then controlling several key events in sperm capacitation, such as membrane and cytoskeleton remodeling. Then, we found different clusters of molecules possibly involved in this pathway and connecting it to the immune system. In conclusion, this work adds a piece to the puzzle of protease and kinase crosstalk involved in the physiology of sperm cells.
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