Development of Rapid and High-Precision Colorimetric Device for Organophosphorus Pesticide Detection Based on Microfluidic Mixer Chip

The excessive pesticide residues in cereals, fruit and vegetables is a big threat to human health, and it is necessary to develop a portable, low-cost and high-precision pesticide residue detection scheme to replace the large-scale laboratory testing equipment for rapid detection of pesticide residues. In this study, a colorimetric device for rapid detection of organophosphorus pesticide residues with high precision based on a microfluidic mixer chip was proposed. The microchannel structure with high mixing efficiency was determined by fluid dynamics simulation, while the corresponding microfluidic mixer chip was designed. The microfluidic mixer chip was prepared by a self-developed liquid crystal display (LCD) mask photo-curing machine. The influence of printing parameters on the accuracy of the prepared chip was investigated. The light source with the optimal wavelength of the device was determined by absorption spectrum measurement, and the relationship between the liquid reservoir depth and detection limit was studied by experiments. The correspondence between pesticide concentration and induced voltage was derived. The minimum detection concentration of the device could reach 0.045 mg·L−1 and the average detection time was reduced to 60 s. The results provide a theoretical and experimental basis for portable and high-precision detection of pesticide residues

Janus kinase inhibitors ruxolitinib and baricitinib impair glycoprotein-VI mediated platelet function

Several Janus kinase (JAK) inhibitors (jakinibs) have recently been approved to treat inflammatory, autoimmune and hematological conditions. Despite emerging roles for JAKs and downstream signal transducer and activator of transcription (STAT) proteins in platelets, it remains unknown whether jakinibs affect platelet function. Here, we profile platelet biochemical and physiological responses in vitro in the presence of five different clinically relevant jakinibs, including ruxolitinib, upadacitinib, oclacitinib, baricitinib and tofacitinib. Flow cytometry, microscopy and other assays found that potent JAK1/2 inhibitors baricitinib and ruxolitinib reduced platelet adhesion to collagen, as well as platelet aggregation, secretion and integrin αIIbβ3 activation in response to the glycoprotein VI (GPVI) agonist collagen-related peptide (CRP-XL). Western blot analysis demonstrated that jakinibs reduced Akt phosphorylation and activation following GPVI activation, where ruxolitinib and baricitinib prevented DAPP1 phosphorylation. In contrast, jakinibs had no effects on platelet responses to thrombin. Inhibitors of GPVI and JAK signaling also abrogated platelet STAT5 phosphorylation following CRP-XL stimulation. Additional pharmacologic experiments supported roles for STAT5 in platelet secretion, integrin activation and cytoskeletal responses. Together, our results demonstrate that ruxolitinib and baricitinib have inhibitory effects on platelet function in vitro and support roles for JAK/STAT5 pathways in GPVI/ITAM mediated platelet function

Potentiation of TRAP-6-induced platelet dense granule release by blockade of P2Y₁₂ signaling with MRS2395

<p>The release of ADP from platelet dense granules and its binding to platelet P2Y₁₂ receptors is key to amplifying the initial hemostatic response and propagating thrombus formation. P2Y₁₂ has thus emerged as a therapeutic target to safely and effectively prevent secondary thrombotic events in patients with acute coronary syndrome or a history of myocardial infarction. Pharmacological inhibition of P2Y₁₂ receptors represents a useful approach to better understand the signaling mediated by these receptors and to elucidate the role of these receptors in a multitude of platelet hemostatic and thrombotic responses. The present work examined and compared the effects of four different P2Y₁₂ inhibitors (MRS2395, ticagrelor, PSB 0739, and AR-C 66096) on platelet function in a series of in vitro studies of platelet dense granule secretion and trafficking, calcium generation, and protein phosphorylation. Our results show that in platelets activated with the PAR-1 agonist TRAP-6 (thrombin receptor-activating peptide), inhibition of P2Y₁₂ with the antagonist MRS2395, but not ticagrelor, PSB 0739 or AR-C 66096, potentiated human platelet dense granule trafficking to the plasma membrane and release into the extracellular space, cytosolic Ca²⁺ influx, and phosphorylation of GSK3β-Ser9 through a PKC-dependent pathway. These results suggest that inhibition of P2Y₁₂ with MRS2395 may act in concert with PAR-1 signaling and result in the aberrant release of ADP by platelet dense granules, thus reducing or counteracting the anticipated anti-platelet efficacy of this inhibitor.</p