Geochemistry and origin of inorganic contaminants in soil, river sediment and surface water in a heavily urbanized river basin.

Understanding the geochemistry and contamination of rivers affected by anthropogenic activities is paramount to water resources management. The Asopos river basin in central Greece is facing environmental quality deterioration threats due to industrial, urban and agricultural activities. Here, the geochemistry of river sediments and adjacent soil in terms of major and trace elements (Al, Ca, Mg, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn) and the geochemical composition of surface water in terms of major ions, trace elements and nutrients along the Asopos river basin were determined. In addition, this study characterized potential nitrate sources through the analysis of stable isotope composition of NO3- (δ15Ν-ΝΟ3- and δ18Ο-ΝΟ3-). Results indicated that specific chemical constituents including nutrients (NO2-, NH4+, PO43-) and major ions (Na+, Cl-) were highest in the urban, industrialized and downstream areas. On the other hand, nitrate (NO3-) concentration in river water (median 7.9 mg/L) showed a decreasing trend from the upstream agricultural sites to the urban area and even more in the downstream of the urban area sites. Ionic ratios (NO3-/Cl-) and δ15Ν-ΝΟ3- values (range from +10.2 ‰ to +15.7 ‰), complemented with a Bayesian isotope mixing model, clearly showed the influence of organic wastes from septic systems and industries operating in the urban area on river nitrate geochemistry. The interpretation of geochemical data of soil and river sediment samples demonstrated the strong influence of local geology on Cr, Fe, Mn and Ni content, with isolated samples showing elevated concentrations of Cd, Cu, Pb and Zn, mostly within the industrialized urban environment. The calculation of enrichment factors based on the national background concentrations provided limited insights into the origin of geogenic metals. Overall, this study highlighted the need for a more holistic approach to assess the impact of the geological background and anthropogenic activities on river waters and sediments.

Mechanosensitive ACKR4 scavenges CCR7 chemokines to facilitate T cell de-adhesion and passive transport by flow in inflamed afferent lymphatics.

T cell migration via afferent lymphatics to draining lymph nodes (dLNs) depends on expression of CCR7 in T cells and CCL21 in the lymphatic vasculature. Once T cells have entered lymphatic capillaries, they slowly migrate into contracting collecting vessels. Here, lymph flow picks up, inducing T cell detachment and rapid transport to the dLNs. We find that the atypical chemokine receptor 4 (ACKR4), which binds and internalizes CCL19 and CCL21, is induced by lymph flow in endothelial cells lining lymphatic collectors, enabling them to scavenge these chemokines. In the absence of ACKR4, migration of T cells to dLNs in TPA-induced inflammation is significantly reduced. While entry into capillaries is not impaired, T cells accumulate in the ACKR4-deficient dermal collecting vessel segments. Overall, our findings identify an ACKR4-mediated mechanism by which lymphatic collectors facilitate the detachment of lymph-borne T cells in inflammation and their transition from crawling to free-flow toward the dLNs.

Transcription factor FOXP2 is a flow-induced regulator of collecting lymphatic vessels.

The lymphatic system is composed of a hierarchical network of fluid absorbing lymphatic capillaries and transporting collecting vessels. Despite distinct functions and morphologies, molecular mechanisms that regulate the identity of the different vessel types are poorly understood. Through transcriptional analysis of murine dermal lymphatic endothelial cells (LECs), we identified Foxp2, a member of the FOXP family of transcription factors implicated in speech development, as a collecting vessel signature gene. FOXP2 expression was induced after initiation of lymph flow in vivo and upon shear stress on primary LECs in vitro. Loss of FOXC2, the major flow-responsive transcriptional regulator of lymphatic valve formation, abolished FOXP2 induction in vitro and in vivo. Genetic deletion of Foxp2 in mice using the endothelial-specific Tie2-Cre or the tamoxifen-inducible LEC-specific Prox1-CreERT2 line resulted in enlarged collecting vessels and defective valves characterized by loss of NFATc1 activity. Our results identify FOXP2 as a new flow-induced transcriptional regulator of collecting lymphatic vessel morphogenesis and highlight the existence of unique transcription factor codes in the establishment of vessel-type-specific endothelial cell identities.

Upregulation of VCAM-1 in lymphatic collectors supports dendritic cell entry and rapid migration to lymph nodes in inflammation.

Dendritic cell (DC) migration to draining lymph nodes (dLNs) is a slow process that is believed to begin with DCs approaching and entering into afferent lymphatic capillaries. From capillaries, DCs slowly crawl into lymphatic collectors, where lymph flow induced by collector contraction supports DC detachment and thereafter rapid, passive transport to dLNs. Performing a transcriptomics analysis of dermal endothelial cells, we found that inflammation induces the degradation of the basement membrane (BM) surrounding lymphatic collectors and preferential up-regulation of the DC trafficking molecule VCAM-1 in collectors. In crawl-in experiments performed in ear skin explants, DCs entered collectors in a CCR7- and ß1 integrin-dependent manner. In vivo, loss of ß1-integrins in DCs or of VCAM-1 in lymphatic collectors had the greatest impact on DC migration to dLNs at early time points when migration kinetics favor the accumulation of rapidly migrating collector DCs rather than slower capillary DCs. Taken together, our findings identify collector entry as a critical mechanism enabling rapid DC migration to dLNs in inflammation.