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Sample preparation under turbulent flow with renewable sorbent† † Electronic supplementary information (ESI) available: A short video demonstrating the bead injection methodology using a transparent tube. The method used in Section 2.6 ‘Proof of concept: application to natural samples’. See DOI: 10.1039/d1ja00252j

ABSTRACT: Turbulent flow chromatography is an online solid phase extraction mode that achieves the extraordinary effect of proxying an upper molecular weight cutoff for the retained molecules, based on loading the sample at high linear velocities. Despite the potential of being a universal sample preparation technique prior to inductively coupled plasma mass spectrometry and liquid chromatography mass spectrometry, it employs specific hardware and expensive consumables. In the present work we apply this technique using off-the-shelf fluidic components and the niche “bead injection” methodology. For the first time, this procedure has been executed with a pressure of approximately 20 bar, compared to the low pressure of the classic setup, achieving a sample throughput >285 h−1 for the SPE/TFC procedure, or 20 h−1 if the procedure involves renewing the sorbent, using no more than 4 mg of sorbent for every μ-SPE. Another novelty is that sorbent packing and unpacking has been controlled with a smart method using real-time pressure feedback as quality control for truly unattended operation. Finally, the turbulent flow chromatography principle has been comprehensively characterized, providing similar performance to that demonstrated in earlier literature, and the ancillary sample preparation capabilities, e.g., in-valve acidification, have been demonstrated by the fractionation of gadolinium in surface waters prior to ICP-MS, an element of increasing surface water concern due to its use as a magnetic resonance contrast agent. An open platform for sample preparation under turbulent flow conditions with stationary phase renewal and smart control.

SUBMITTER: Cocovi-Solberg D

PROVIDER: S-EPMC8565614 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Vascular endothelial and hematopoietic cells hold a close relationship that is initiated during development, but that continuously broadens to include new functions aimed at maintaining homeostasis in the adult organism. Here, we report on a population of aortic intimal resident macrophages (MacAIRs) that is a permanent feature of the inner vessel wall (tunica intima) of the aorta in regions of turbulent flow and that shares the luminal surface with the endothelium. MacAIRs were found to be transcriptionally distinct from other macrophages and closely associated with the endothelium in the absence of pathology. Lineage tracing analysis indicated that adult MacAIRs were derived from definitive hematopoietic lineage precursors that migrated from the ductus arteriosus seeding the aorta immediately post-birth in areas of turbulent flow including the lesser curvature of the aortic arch and branch openings. These aortic resident macrophages expanded via direct cell renewal and continued to grow in aged aortae to also populate regions of laminar flow. Binding of MacAIRs relied heavily on expression of endothelial ICAM that interacts with CD11c on macrophages. Genetic deletion of CD11c significantly impaired anchorage of MacAIRs to the endothelium. To clarify the biological significance of this population of macrophages, we used a dual diphtheria toxin induced-depletion system that efficiently eliminated MacAIRs in adult mice. Utilizing this model, we found that absence of MacAIRs led to progressive fibrin accumulation and formation of microclots that, once dislodged, can cause blockade of vessels and organ failure. In fact, 40% of the animals with genetic-mediated depletion of MacAIRs died within 2 weeks post-depletion suggesting that these macrophages were required to clear fibrin deposits in regions of turbulent blood flow. These findings advance our knowledge of how vascular endothelial cells profit from this unique interaction with macrophages to control hemostasis, prevent intravascular clotting, and ensure vascular health.

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