Description: Aims Retention of low-density lipoprotein (LDL) cholesterol beneath the arterial endothelium initiates an inflammatory response culminating in atherosclerosis. Since the overlying endothelium is healthy and intact early on, it is likely that LDL passes through endothelial cells by transcytosis. However, technical challenges have made confirming this notion and elucidating the mechanisms of transcytosis difficult. We developed a novel assay for measuring LDL transcytosis in real time across coronary endothelial cell monolayers; we used this approach to identify the receptor involved. Methods and results Murine aortas were perfused ex vivo with LDL and dextran of a smaller molecular radius. LDL (but not dextran) accumulated under the endothelium, indicating that LDL transcytosis occurs in intact vessels. We then confirmed that LDL transcytosis occurs in vitro using human coronary artery endothelial cells. An assay was developed to quantify transcytosis of DiI-LDL in real time using total internal reflection fluorescence microscopy. DiI-LDL transcytosis was inhibited by excess unlabelled LDL, while degradation of the LDL receptor by PCSK9 had no effect. Instead, LDL colocalized partially with the scavenger receptor SR-BI and overexpression of SR-BI increased LDL transcytosis; knockdown by siRNA significantly reduced it. Excess HDL, the canonical SR-BI ligand, significantly decreased LDL transcytosis. Aortas from SR-BI -deficient mice were perfused ex vivo with LDL and accumulated significantly less sub-endothelial LDL compared with wild-type littermates. Conclusion We developed an assay to quantify LDL transcytosis across endothelial cells and discovered an unexpected role for SR-BI. Elucidating the mechanisms of LDL transcytosis may identify novel targets for the prevention or therapy of atherosclerosis.

Description: We have investigated the seismic anisotropy beneath the Central Andean southern Puna plateau by applying shear wave splitting analysis and shear wave splitting tomography to local S waves and teleseismic SKS, SKKS and PKS phases. Overall, a very complex pattern of fast directions throughout the southern Puna plateau region and a circular pattern of fast directions around the region of the giant Cerro Galan ignimbrite complex are observed. In general, teleseismic lag times are much greater than those for local events which are interpreted to reflect a significant amount of sub and inner slab anisotropy. The complex pattern observed from shear wave splitting analysis alone is the result of a complex 3-D anisotropic structure under the southern Puna plateau. Our application of shear wave splitting tomography provides a 3-D model of anisotropy in the southern Puna plateau that shows different patterns depending on the driving mechanism of upper-mantle flow and seismic anisotropy. The trench parallel a -axes in the continental lithosphere above the slab east of 68W may be related to deformation of the overriding continental lithosphere since it is under compressive stresses which are orthogonal to the trench. The more complex pattern below the Cerro Galan ignimbrite complex and above the slab is interpreted to reflect delamination of continental lithosphere and upwelling of hot asthenosphere. The a -axes beneath the Cerro Galan, Cerro Blanco and Carachi Pampa volcanic centres at 100 km depth show some weak evidence for vertically orientated fast directions, which could be due to vertical asthenospheric flow around a delaminated block. Additionally, our splitting tomographic model shows that there is a significant amount of seismic anisotropy beneath the slab. The subslab mantle west of 68W shows roughly trench parallel horizontal a -axes that are probably driven by slab roll back and the relatively small coupling between the Nazca slab and the underlying mantle. In contrast, the subslab region (i.e. depths greater than 200 km) east of 68W shows a circular pattern of a -axes centred on a region with small strength of anisotropy (Cerro Galan and its eastern edge) which suggest the dominant mechanism is a combination of slab roll back and flow driven by an overlying abnormally heated slab or possibly a slab gap. There seems to be some evidence for vertical flow below the slab at depths of 200–400 km driven by the abnormally heated slab or slab gap. This cannot be resolved by the tomographic inversion due to the lack of ray crossings in the subslab mantle.

Description: During inflammation, leukocyte–endothelial cell interactions generate molecular signals that regulate cell functions. The Ca 2+ - and F-actin–binding leukocyte-specific protein 1 (LSP1) expressed in leukocytes and nonhematopoietic endothelial cells is pivotal in regulating microvascular permeability and leukocyte recruitment. However, cell-specific function of LSP1 during leukocyte recruitment remains elusive. Using intravital microscopy of cremasteric microvasculature of chimeric LSP1-deficient mice, we show that not neutrophil but endothelial LSP1 regulates neutrophil transendothelial migration and extravascular directionality without affecting the speed of neutrophil migration in tissue in response to CXCL2 chemokine gradient. The expression of PECAM-1–sensitive α 6 β 1 integrins on the surface of transmigrated neutrophils was blunted in mice deficient in endothelial LSP1. Functional blocking studies in vivo and in vitro elucidated that α 6 β 1 integrins orchestrated extravascular directionality but not the speed of neutrophil migration. In LSP1-deficient mice, PECAM-1 expression was reduced in endothelial cells, but not in neutrophils. Similarly, LSP1-targeted small interfering RNA silencing in murine endothelial cells mitigated mRNA and protein expression of PECAM-1, but not ICAM-1 or VCAM-1. Overexpression of LSP1 in endothelial cells upregulated PECAM-1 expression. Furthermore, the expression of transcription factor GATA-2 that regulates endothelial PECAM-1 expression was blunted in LSP1-deficient or LSP1-silenced endothelial cells. The present study unravels endothelial LSP1 as a novel cell-specific regulator of integrin α 6 β 1 -dependent neutrophil extravascular chemotactic function in vivo, effective through GATA-2–dependent transcriptional regulation of endothelial PECAM-1 expression.

Description: Obesity is associated with inflammation and immune cell recruitment to adipose tissue, muscle and intima of atherosclerotic blood vessels. Obesity and hyperlipidemia are also associated with tissue insulin resistance and can compromise insulin delivery to muscle. The muscle/fat microvascular endothelium mediates insulin delivery and facilitates monocyte transmigration, yet its contribution to the consequences of hyperlipidemia is poorly understood. Using primary endothelial cells from human adipose tissue microvasculature (HAMEC), we investigated the effects of physiological levels of fatty acids on endothelial inflammation and function. Expression of cytokines and adhesion molecules was measured by RT-qPCR. Signaling pathways were evaluated by pharmacological manipulation and immunoblotting. Surface expression of adhesion molecules was determined by immunohistochemistry. THP1 monocyte interaction with HAMEC was measured by cell adhesion and migration across transwells. Insulin transcytosis was measured by total internal reflection fluorescence microscopy. Palmitate, but not palmitoleate, elevated the expression of IL-6, IL-8, TLR2 (Toll-like receptor 2), and intercellular adhesion molecule 1 (ICAM-1). HAMEC had markedly low fatty acid uptake and oxidation, and CD36 inhibition did not reverse the palmitate-induced expression of adhesion molecules, suggesting that inflammation did not arise from palmitate uptake/metabolism. Instead, inhibition of TLR4 to NF-B signaling blunted palmitate-induced ICAM-1 expression. Importantly, palmitate-induced surface expression of ICAM-1 promoted monocyte binding and transmigration. Conversely, palmitate reduced insulin transcytosis, an effect reversed by TLR4 inhibition. In summary, palmitate activates inflammatory pathways in primary microvascular endothelial cells, impairing insulin transport and increasing monocyte transmigration. This behavior may contribute in vivo to reduced tissue insulin action and enhanced tissue infiltration by immune cells.

Description: The Bransfield Basin is a back-arc basin located in Western Antarctica between the South Shetland Islands and Antarctic Peninsula. Although the subduction of the Phoenix plate under the South Shetland block has ceased, extension continues through a combination of slab rollback and transtensional motions between the Scotia and Antarctic plates. This process has created a continental rift in the basin, interleaved with volcanic islands and seamounts, which may be near the transition from rifting to seafloor spreading. In the framework of the BRAVOSEIS project (2017–2020), we deployed a dense amphibious seismic network in the Bransfield Strait comprising 15 land stations and 24 ocean-bottom seismometers, as well as a network of 6 moored hydrophones; and acquired marine geophysics data including multibeam bathymetry, sub-bottom profiler, gravity & mag-netics, multi-channel seismics, and seismic refraction data. The experiment has collected a unique, high quality, and multifaceted geophysical data set in the Central Bransfield Basin, with a special focus on Orca and Humpback seamounts. Preliminary results confirm that the Bransfield region has slab-related intermediate depth seismicity, with earthquake characteristics suggesting distributed extension across the rift. Gravity and magnetic highs delineate a segmented rift with along-axis variations that are consistent with increased accumulated strain to the northeast. Orca volcano shows evidences of an active caldera and magma accumulation at shallow depths, while Humpback volcano has evolved past the caldera stage and is currently dominated by rifting structures. These differences suggest that volcanic evolution is influenced by the position along the rift. Although a lot of analysis remains, these results provide useful constraints on the structure and dynamics of the Bransfield rift and asso-ciated volcanoes.

Description: Northwestern Namibia, at the landfall of the Walvis Ridge, was affected by the Tristan da Cunha mantle plume during continental rupture between Africa and South America, as evidenced by the presence of the Etendeka continental flood basalts. Here we use data from a passive-source seismological network to investigate the upper mantle structure and to elucidate the Cretaceous mantle plume-lithosphere interaction. Receiver functions reveal an interface associated with a negative velocity contrast within the lithosphere at an average depth of 80 km. We interpret this interface as the relic of the lithosphereasthenosphere boundary (LAB) formed during the Mesozoic by interaction of the Tristan da Cunha plume head with the pre-existing lithosphere. The velocity contrast might be explained by stagnated and ‘‘frozen’’ melts beneath an intensively depleted and dehydrated peridotitic mantle. The present-day LAB is poorly visible with converted waves, indicating a gradual impedance contrast. Beneath much of the study area, converted phases of the 410 and 660 km mantle transition zone discontinuities arrive 1.5 s earlier than in the landward plume-unaffected continental interior, suggesting high velocities in the upper mantle caused by a thick lithosphere. This indicates that after lithospheric thinning during continental breakup, the lithosphere has increased in thickness during the last 132 Myr. Thermal cooling of the continental lithosphere alone cannot produce the lithospheric thickness required here. We propose that the remnant plume material, which has a higher seismic velocity than the ambient mantle due to melt depletion and dehydration, significantly contributed to the thickening of the mantle lithosphere.

Description: The amphibian Walvis Ridge Passive-Source Seismic Experiment (WALPASS) have been operated for a period of two years from 2010 to 2012 in the area where the Walvis Ridge intersects the continental margin of northwestern Namibia. The deployment was intended to study the lithospheric and upper mantle structure in the ocean-continent transition area beneath the passive continental margin. The main idea is to find seismic anomalies related to the postulated hotspot track from the continent to the ocean along the Walvis Ridge that links the Etendeka continental flood-basalt province to the Tristan da Cunha hotspot in the mid Atlantic ocean. This could provide clues of the role of plume-lithosphere interaction during the continental break-up. We present here seismic structures of the crustal and mantle lithosphere in this geophysically little studied region using seismic methods including P and S receiver functions and shear wave splitting. The average crustal thickness in the continental Namibia is ~35 km with a relatively low Vp/Vs ratio of 1.7. Underneath the NE extension of the Walvis Ridge the crust is the thickest (45 km) with a high Vp/Vs ratio (〉1.80). The thick crust and high Vp/Vs ratio beneath the Walvis Ridge are consistent with high Vp derived by controlled source seismics, implying a magmatic underplating. A low velocity zone in the mantle is observed at depths of 60-120 km, possibly representing the base of the lithosphere. The P-to-S converted phases at the 410 and 660 km discontinuities arrive 2-3 s earlier, indicating higher upper mantle velocities (+5%). Seismic anisotropy in the mantle derived by the SKS splitting exhibits a pattern of the plume and plate interaction.

Description: Passive continental margins offer the unique opportunity to study the processes involved in continental extension and break up and the role of hot-spot related magmatism. Several geophysical experiments (Seismics, Magnetotellurics and Seismology) were therefore carried out 2011 and 2012 in northern Namibia to image crust and upper mantle at the landfall of the Walvis Ridge. The aim of these studies is to shed light on the present-day structure of this area to understand the dynamics of the breakup of Pangaea and the processes involved. First results of the seismic part of the project show anomalous velocity structures and reflective properties in the mid- and lower crust. The lower crust (onshore) is characterized by an unusual high velocity body which might be associated with magmatic processes of the plume continent interaction. The distribution of the upper mantle wave propagation velocities shows a rather complex, very inhomogeneous pattern. Onshore magnetotelluric (MT) data were acquired at 167 sites in a ~140 km wide and ~260 km long corridor from the Atlantic Ocean through the Kaoko Mobile Belt onto the Congo Craton. The data are generally of excellent quality. A first inspection of magnetotelluric and vertical magnetic transfer functions indicates significant three-dimensional (3-D) structures in the crust and upper mantle, particularly in the Western Kaoko Zone in the vicinity of prominent shear zones. The seismological team operates a passive-source seismic experiment for two years onshore/offshore NW Namibia. The seismic network consists of 28 land-based and 12 ocean-bottom stations covering an area of 400km x 800km. Different seismic methods, such as body wave and surface wave tomography, receiver function, shear wave splitting, etc, will be used to image the seismic anomalies in the upper mantle and to map the thickness of the crust and mantle lithosphere in this ocean-continental transition area. The aim is to find the mantle deformation styles related to the plume-lithosphere interaction along the Walvis Ridge.

Description: The Walvis Ridge (WR) is the most prominent hotspot track related to the opening in the South Atlantic Ocean. Several hypotheses have been developed to explain its origin and evolution. The presence of a massive magmatic structure at the landfall of the WR in Northwest Namibia raised speculation about the role of a hotspot during the opening of the South Atlantic ocean. To investigate its deeper velocity structure at the junction of the WR with the African continent was the focus of the amphibious seismological WALPASS experiment. In total 12 oceanbottom seismometers and 28 broad-band land stations were installed between 2010 and 2012 to acquire seismological data. Here,we present the results of seismic ambient noise tomography to investigate to which extent the Tristan hotspot modified the crustal structure in the landward prolongation of the ridge and in the adjacent oceanic basins. For the tomography, vertical and hydrophone component cross correlations for 〉300 d for OBS stations and between 1 and 2 yr for land stations data were analysed. More than 49 000 velocity measurements (742 dispersion curves) were inverted for group velocity maps at 75 individual signal periods, which then had been inverted for a regional 3-D shear wave velocity model. The resulting 3-D model reveals structural features of the crust related to the continent–ocean transition and its disturbance caused by the initial formation of the WR ∼130 Ma. We found relatively thick continental crust below Northwest Namibia and below the near-shore part of the WR, a strong asymmetry offshore with typical, thin oceanic crust in the Namibe Basin (crossing over into the Angola Basin further offshore) to the North and a wide zone of transitional crust towards the Walvis Basin south of the WR.