Neuropilins 1 and 2 mediate neointimal hyperplasia and re-endothelialization following arterial injury

AIMS: Neuropilins 1 and 2 (NRP1 and NRP2) play crucial roles in endothelial cell migration contributing to angiogenesis and vascular development. Both NRPs are also expressed by cultured vascular smooth muscle cells (VSMCs) and are implicated in VSMC migration stimulated by PDGF-BB, but it is unknown whether NRPs are relevant for VSMC function in vivo. We investigated the role of NRPs in the rat carotid balloon injury model, in which endothelial denudation and arterial stretch induce neointimal hyperplasia involving VSMC migration and proliferation. METHODS AND RESULTS: NRP1 and NRP2 mRNAs and proteins increased significantly following arterial injury, and immunofluorescent staining revealed neointimal NRP expression. Down-regulation of NRP1 and NRP2 using shRNA significantly reduced neointimal hyperplasia following injury. Furthermore, inhibition of NRP1 by adenovirally overexpressing a loss-of-function NRP1 mutant lacking the cytoplasmic domain (ΔC) reduced neointimal hyperplasia, whereas wild-type (WT) NRP1 had no effect. NRP-targeted shRNAs impaired, while overexpression of NRP1 WT and NRP1 ΔC enhanced, arterial re-endothelialization 14 days after injury. Knockdown of either NRP1 or NRP2 inhibited PDGF-BB-induced rat VSMC migration, whereas knockdown of NRP2, but not NRP1, reduced proliferation of cultured rat VSMC and neointimal VSMC in vivo. NRP knockdown also reduced the phosphorylation of PDGFα and PDGFβ receptors in rat VSMC, which mediate VSMC migration and proliferation. CONCLUSION: NRP1 and NRP2 play important roles in the regulation of neointimal hyperplasia in vivo by modulating VSMC migration (via NRP1 and NRP2) and proliferation (via NRP2), independently of the role of NRPs in re-endothelialization

VEGF (Vascular Endothelial Growth Factor) Induces NRP1 (Neuropilin-1) Cleavage via ADAMs (a Disintegrin and Metalloproteinase) 9 and 10 to Generate Novel Carboxy- Terminal NRP1 Fragments That Regulate Angiogenic Signaling

OBJECTIVE: NRP1(neuropilin-1) acts as a coreceptor for VEGF (vascular endothelial growth factor) with an essential role in angiogenesis. Recent findings suggest that posttranslational proteolytic cleavage of VEGF receptors may be an important mechanism for regulating angiogenesis, but the role of NRP1 proteolysis and the NRP1 species generated by cleavage in endothelial cells is not known. To characterize NRP1 proteolytic cleavage in endothelial cells, determine the mechanism, and investigate the role of NRP1 cleavage in regulation of endothelial cell function. APPROACH AND RESULTS: NRP1 species comprising the carboxy (C)-terminal and transmembrane NRP1 domains but lacking the ligand-binding A and B regions are constitutively expressed in endothelial cells. Generation of these C-terminal domain NRP1 proteins is upregulated by phorbol ester and Ca2+ ionophore, and reduced by pharmacological inhibition of metalloproteinases, by small interfering RNA-mediated knockdown of 2 members of ADAM (a disintegrin and metalloproteinase) family, ADAMs 9 and 10, and by a specific ADAM10 inhibitor. Furthermore, VEGF upregulates expression of these NRP1 species in an ADAM9/10-dependent manner. Transduction of endothelial cells with adenoviral constructs expressing NRP1 C-terminal domain fragments inhibited VEGF-induced phosphorylation of VEGFR2 (VEGF receptor tyrosine kinase)/KDR and decreased VEGF-stimulated endothelial cell motility and angiogenesis in coculture and aortic ring sprouting assays. CONCLUSIONS: These findings identify novel NRP1 species in endothelial cells and demonstrate that regulation of NRP1 proteolysis via ADAMs 9 and 10 is a new regulatory pathway able to modulate VEGF angiogenic signaling

Renal tubular absorption of β2 microglobulin

Renal tubular absorption of β2 microglobulin. 125Iodinated human β2 microglobulin (β2m, 5 to 30 mg) was administered to anesthetized rats. Clearance studies showed a low threshold of excretion of injected β2m and a high Tm of 400 to 600 µg · min-1 · kg-1. A glomerular sieving coefficient of 0.97 was calculated as the slope of the curve: β2m excretion rate = F (plasma β2m × glomerular filtration rate) for values above saturation. Electrophoresis analysis of proteinuria in agarose gel and sodium dodecyl sulfate polyacrylamide gel showed that injection of saturating doses of β2m induced the excretion of proteins of similar size but different charge and that of other proteins of different size. Among the latter, some were excreted transiently in association with β2m, whereas others had a delayed excretion suggesting existence of a complex mechanism of reabsorption whose steps remain to be elucidated.Absorption tubulaire rénale de la β2 microglobuline. De 5 à30 mg de β2 microglobuline (β2m) humaine marquée à l'Iode 125 ont été injectés à des rats anesthésiés. Des études de clairance ont montré un seuil d'excrétion bas et un Tm élevé de 400 à 600 µg · min-1 · kg-1. Un coefficient de tamisage de 0,97 a été mesuré à partir de la pente de la courbe: excretion de β2m = F (concentration plasmatique de β2m × filtration glomérulaire) pour les points au-dessus de la saturation. L'analyse de la protéinurie par électrophorèse sur gel d'agarose et sur gel de polyacrylamide avec dodecyl-sulfate de sodium a montré que l'injection de doses saturantes de β2m provoque l'excretion de protéines de même taille mais de charge différente, ainsi que de protéines de taille différente. Parmi ces dernières, certaines sont excrétées de manière transitoire et en même temps que la β2m, tandis que d'autres ont une excrétion retardée suggérant l'existence d'un mécanisme de réabsorption complexe dont les étapes restent à étudier

Integrating multiple satellite observations into a coherent dataset to monitor the full water cycle – application to the Mediterranean region

The Mediterranean region is one of the climate hotspots where the climate change impacts are both pronounced and documented. The HyMeX (Hydrometeorological Mediterranean eXperiment) aims to improve our understanding of the water cycle from the meteorological to climate scales. However, monitoring the water cycle with Earth observations (EO) is still a challenge: EO products are multiple, and their utility is degraded by large uncertainties and incoherences among the products. Over the Mediterranean region, these difficulties are exacerbated by the coastal/mountainous regions and the small size of the hydrological basins. Therefore, merging/integration techniques have been developed to reduce these issues. We introduce here an improved methodology that closes not only the terrestrial but also the atmospheric and ocean budgets. The new scheme allows us to impose a spatial and temporal multi-scale budget closure constraint. A new approach is also proposed to downscale the results from the basin to pixel scales (at the resolution of 0.25∘). The provided Mediterranean WC budget is, for the first time, based mostly on observations such as the GRACE water storage or the netflow at the Gibraltar Strait. The integrated dataset is in better agreement with in situ measurements, and we are now able to estimate the Bosporus Strait annual mean netflow.</p

High-resolution imaging of compact high-velocity clouds (II)

We have imaged five compact high-velocity clouds in HI with arcmin angular- and km/s spectral-resolution using the WSRT. Supplementary total-power data, which is fully sensitive to both the cool and warm components of HI, is available for comparison for all the sources, albeit with angular resolutions that vary from 3' to 36'. The fractional HI flux in compact CNM components varies from 4% to 16% in our sample. All objects have at least one local peak in the CNM column which exceeds about 10^19 cm^-2 when observed with arcmin resolution. It is plausible that a peak column density of 1-2x10^19 cm^-2 is a prerequisite for the long-term survival of these sources. One object in our sample, CHVC120-20-443 (Davies' cloud), lies in close projected proximity to the disk of M31. This object is characterized by exceptionally broad linewidths in its CNM concentrations (more than 5 times greater than the median value). These CNM concentrations lie in an arc on the edge of the source facing the M31 disk, while the diffuse HI component of this source has a position offset in the direction of the disk. All of these attributes suggest that CHVC120-20-443 is in a different evolutionary state than most of the other CHVCs which have been studied. Similarly broad CNM linewidths have only been detected in one other object, CHVC111-07-466, which also lies in the Local Group barycenter direction and has the most extreme radial velocity known. A distinct possibility for Davies' cloud seems to be physical interaction of some type with M31. The most likely form of this interaction might be the ram-pressure or tidal- stripping by either one of M31's visible dwarf companions, M32 or NGC205, or else by a dark companion with an associated HI condensation.Comment: 12 pages, 11 (low res.) png figs, accepted for pub. in A&

Superbubble evolution including the star-forming clouds: Is it possible to reconcile LMC observations with model predictions?

Here we present a possible solution to the apparent discrepancy between the observed properties of LMC bubbles and the standard, constant density bubble model. A two-dimensional model of a wind-driven bubble expanding from a flattened giant molecular cloud is examined. We conclude that the expansion velocities derived from spherically symmetric models are not always applicable to elongated young bubbles seen almost face-on due to the LMC orientation. In addition, an observational test to differentiate between spherical and elongated bubbles seen face-on is discussed.Comment: 25 pages, 7 figures, accepted to ApJ (September, 1999 issue

Hepatitis C virus infection protein network

A proteome-wide mapping of interactions between hepatitis C virus (HCV) and human proteins was performed to provide a comprehensive view of the cellular infection. A total of 314 protein–protein interactions between HCV and human proteins was identified by yeast two-hybrid and 170 by literature mining. Integration of this data set into a reconstructed human interactome showed that cellular proteins interacting with HCV are enriched in highly central and interconnected proteins. A global analysis on the basis of functional annotation highlighted the enrichment of cellular pathways targeted by HCV. A network of proteins associated with frequent clinical disorders of chronically infected patients was constructed by connecting the insulin, Jak/STAT and TGFβ pathways with cellular proteins targeted by HCV. CORE protein appeared as a major perturbator of this network. Focal adhesion was identified as a new function affected by HCV, mainly by NS3 and NS5A proteins

ViralORFeome: an integrated database to generate a versatile collection of viral ORFs

Large collections of protein-encoding open reading frames (ORFs) established in a versatile recombination-based cloning system have been instrumental to study protein functions in high-throughput assays. Such ‘ORFeome’ resources have been developed for several organisms but in virology, plasmid collections covering a significant fraction of the virosphere are still needed. In this perspective, we present ViralORFeome 1.0 (http://www.viralorfeome.com), an open-access database and management system that provides an integrated set of bioinformatic tools to clone viral ORFs in the Gateway® system. ViralORFeome provides a convenient interface to navigate through virus genome sequences, to design ORF-specific cloning primers, to validate the sequence of generated constructs and to browse established collections of virus ORFs. Most importantly, ViralORFeome has been designed to manage all possible variants or mutants of a given ORF so that the cloning procedure can be applied to any emerging virus strain. A subset of plasmid constructs generated with ViralORFeome platform has been tested with success for heterologous protein expression in different expression systems at proteome scale. ViralORFeome should provide our community with a framework to establish a large collection of virus ORF clones, an instrumental resource to determine functions, activities and binding partners of viral proteins

Multifunctional molecular modulators for perovskite solar cells with over 20% efficiency and high operational stability

Perovskite solar cells present one of the most prominent photovoltaic technologies, yet their stability, scalability, and engineering at the molecular level remain challenging. We demonstrate a concept of multifunctional molecular modulation of scalable and operationally stable perovskite solar cells that exhibit exceptional solar-to-electric power conversion efficiencies. The judiciously designed bifunctional molecular modulator SN links the mercapto-tetrazolium (S) and phenylammonium (N) moieties, which passivate the surface defects, while displaying a structure-directing function through interaction with the perovskite that induces the formation of large grain crystals of high electronic quality of the most thermally stable formamidinium cesium mixed lead iodide perovskite formulation. As a result, we achieve greatly enhanced solar cell performance with efficiencies exceeding 20% for active device areas above 1 cm(2) without the use of antisolvents, accompanied by outstanding operational stability under ambient conditions