Synergistic Activation of ENaC by Three Membrane-bound Channel-activating Serine Proteases (mCAP1, mCAP2, and mCAP3) and Serum- and Glucocorticoid-regulated Kinase (Sgk1) in Xenopus Oocytes

Sodium balance is maintained by the precise regulation of the activity of the epithelial sodium channel (ENaC) in the kidney. We have recently reported an extracellular activation of ENaC-mediated sodium transport (INa) by a GPI-anchored serine protease (mouse channel–activating protein, mCAP1) that was isolated from a cortical collecting duct cell line derived from mouse kidney. In the present study, we have identified two additional membrane-bound serine proteases (mCAP2 and mCAP3) that are expressed in the same cell line. We show that each of these proteases is able to increase INa 6–10-fold in the Xenopus oocyte expression system. INa and the number (N) of channels expressed at the cell surface (measured by binding of a FLAG monoclonal I125-radioiodinated antibody) were measured in the same oocyte. Using this assay, we show that mCAP1 increases INa 10-fold (P < 0.001) but N remained unchanged (P = 0.9), indicating that mCAP1 regulates ENaC activity by increasing its average open probability of the whole cell (wcPo). The serum- and glucocorticoid-regulated kinase (Sgk1) involved in the aldosterone-dependent signaling cascade enhances INa by 2.5-fold (P < 0.001) and N by 1.6-fold (P < 0.001), indicating a dual effect on N and wcPo. Compared with Sgk1 alone, coexpression of Sgk1 with mCAP1 leads to a ninefold increase in INa (P < 0.001) and 1.3-fold in N (P < 0.02). Similar results were observed for mCAP2 and mCAP3. The synergism between CAPs and Sgk1 on INa was always more than additive, indicating a true potentiation. The synergistic effect of the two activation pathways allows a large dynamic range for ENaC-mediated sodium regulation crucial for a tight control of sodium homeostasis

Heterogeneity in tribologically transformed structure (TTS) of Ti-6Al-4V under fretting

Fretting wear is a surface degradation process caused by oscillatory motion and contact slipping. During gross slip, high local stresses and plastic deformation in the surface and subsurface can lead to the creation of a nanosized grained structure called Tribologically Transformed Structure (TTS). The current paper studies the formation of TTS in an alpha-beta Ti-6Al-4V alloy under fretting loading while changing the contact pressure and the number of fretting cycles.Cross-sections of wear scars are observed after polishing and chemical etching. Above a threshold pressure of 300 MPa, TTS appears early in the contact (before 1000 cycles) along with two other structures: a Third Body Layer (TBL) made of compacted debris and a General Deformed Layer (GDL) which is the plastic zone under the TTS. TTS first appears as islands and merges in the middle of the contact after enough cycles. Below 200 MPa, only TBL and GDL are formed. At 200 MPa, only small, localized TTS is found. All structures have the same chemical compositions as the initial bulk material except for the nitrided TBL. TTS has a very high hardness compared to the bulk. TTS was carefully extracted using a Focused Ion Beam (FIB) and its microstructure was observed with a Transmission Electron Microscope (TEM). It shows extreme grain refinement and is composed of two alternated zones. The first zone I is composed of $\alpha$ grains with a size of 20 to 50 nm with crystallographic texture. Zone II comprises nanosized equiaxed grains whose sizes range from 5 to 20 nm without texture. The results made it possible to establish a scenario of the appearance of the TTS according to the conditions of contact pressure and number of fretting cycles

Protease Modulation of the Activity of the Epithelial Sodium Channel Expressed in Xenopus Oocytes

We have investigated the effect of extracellular proteases on the amiloride-sensitive Na+ current (INa) in Xenopus oocytes expressing the three subunits α, β, and γ of the rat or Xenopus epithelial Na+ channel (ENaC). Low concentrations of trypsin (2 μg/ml) induced a large increase of INa within a few minutes, an effect that was fully prevented by soybean trypsin inhibitor, but not by amiloride. A similar effect was observed with chymotrypsin, but not with kallikrein. The trypsin-induced increase of INa was observed with Xenopus and rat ENaC, and was very large (∼20-fold) with the channel obtained by coexpression of the α subunit of Xenopus ENaC with the β and γ subunits of rat ENaC. The effect of trypsin was selective for ENaC, as shown by the absence of effect on the current due to expression of the K+ channel ROMK2. The effect of trypsin was not prevented by intracellular injection of EGTA nor by pretreatment with GTP-γS, suggesting that this effect was not mediated by G proteins. Measurement of the channel protein expression at the oocyte surface by antibody binding to a FLAG epitope showed that the effect of trypsin was not accompanied by an increase in the channel protein density, indicating that proteolysis modified the activity of the channel present at the oocyte surface rather than the cell surface expression. At the single channel level, in the cell-attached mode, more active channels were observed in the patch when trypsin was present in the pipette, while no change in channel activity could be detected when trypsin was added to the bath solution around the patch pipette. We conclude that extracellular proteases are able to increase the open probability of the epithelial sodium channel by an effect that does not occur through activation of a G protein-coupled receptor, but rather through proteolysis of a protein that is either a constitutive part of the channel itself or closely associated with it

Antiretroviral-naive and -treated HIV-1 patients can harbour more resistant viruses in CSF than in plasma

Objectives The neurological disorders in HIV-1-infected patients remain prevalent. The HIV-1 resistance in plasma and CSF was compared in patients with neurological disorders in a multicentre study. Methods Blood and CSF samples were collected at time of neurological disorders for 244 patients. The viral loads were >50 copies/mL in both compartments and bulk genotypic tests were realized. Results On 244 patients, 89 and 155 were antiretroviral (ARV) naive and ARV treated, respectively. In ARV-naive patients, detection of mutations in CSF and not in plasma were reported for the reverse transcriptase (RT) gene in 2/89 patients (2.2%) and for the protease gene in 1/89 patients (1.1%). In ARV-treated patients, 19/152 (12.5%) patients had HIV-1 mutations only in the CSF for the RT gene and 30/151 (19.8%) for the protease gene. Two mutations appeared statistically more prevalent in the CSF than in plasma: M41L (P = 0.0455) and T215Y (P = 0.0455). Conclusions In most cases, resistance mutations were present and similar in both studied compartments. However, in 3.4% of ARV-naive and 8.8% of ARV-treated patients, the virus was more resistant in CSF than in plasma. These results support the need for genotypic resistance testing when lumbar puncture is performe

Glossaire illustré sur les formes d’altération de la pierre

The ICOMOS International Scientific Committee for Stone (ISCS) is providing a forum for the interchange of experience, ideas, and knowledge in the field of stone conservation. ISCS aims at facilitating the publication, dissemination and presentation of state of the art reviews on pre-identified issues. Simplification and demystification of scientific information for practitioners are also part of the main goals of the group. In studies on stone deterioration and conservation, terminological confusions lead to major communication problems between scientists, conservators and practitioners. In this context, it is of primary importance to set up a common language; if degradation patterns can be shown, named and described, then they can be recognised and compared with similar ones in a more accurate way in further investigations. The ISCS glossary constitutes an important tool for scientific discussions on decay phenomena and processes. It is also an excellent basis for tutorials on stone deterioration. It is based on the careful examination of pre-existing glossaries of English terms. It does not aim at replacing these glossaries, often set up originally in a language other than English, and for most of them done to a high standard. As President of ICOMOS I would like to congratulate the International Scientific Committee for Stone and its President Véronique Verges-Belmin for the results of years of research presented in this publication. Stone conservation is a crucial topic in monument conservation and many of our National Committees all over the world hope for advice and help from the specialists familiar with traditional and modern methods of conservation. The Illustrated Glossary on Stone Deterioration Patterns offers a wide range of suggestions and practical advice. Probably, after the English-French version becomes available the Glossary will also be translated into other languages. In view of the accelerating decay of our stone monuments worldwide this is an exemplary contribution which will promote the international cooperation so important in this field.peer-reviewe

Assessment of woodland plant diversity along a rural-urban gradient, implications for urban green infrastructure management and planning.

The increase of urbanization during the last decades (and the related losses of biodiversity) has resulted in a need to consider biodiversity conservation in urban areas. The aim of this paper is to better understand the relations between plant biodiversity, management intensity and landscape planning by comparing several results obtained in cities of western France. First, we compare three urban greenspaces presenting a large management intensity gradient. These observations show that large urban parks with low management intensity can constitute a local "hotspot of biodiversity", even if it is not their management target. The second part focuses on isolated woodlands and aim to assess the response plant species richness to urbanization. A global pattern of increase in species richness is observed along the rural-urban gradient, but contrasted responses occur according to the origin (native or not) of species. Urban woodlands are richer in exotic species but native and forest species are dominant in all woodlands. Spatiotemporal distribution of areas with trees along an urban-rural gradient (so not only woodland, but all tree cover that can be seen by aerial photograph) is analyzed in the third part. Two dates (1958 and 2001) are compared. The total surface area and total number of tree patches did not change between 1958 and 2001. However, the three classes (woodlands, hedges and treelines & isolated trees) of area evolved differently. The areas of semi-natural tree cover (sum of woodlands and hedges) appear quite similar for rural and recently urbanized district (about 8%), in opposition to more ancient urban district where they are almost absent. Thus, such spatio-temporal urban-rural gradient allow considering changes in landscape planning practices. This work contributes to a better understanding of the response of plant species to urbanization and could improve the integration of woodlands in urban planning and management

Contribution of woodlands to biodiversity conservation in urban areas and management implications

Contribution of woodlands to biodiversity conservation in urban areas and management implication

Effect of urban-rural gradient on the distribution of woodland flora

Effect of urban-rural gradient on the distribution of woodland flor

Plant species response to urbanization: comparison of isolated woodland patches in two cities of North-Western France

FR2116International audienceThe effect of urbanization on species distribution has been extensively documented, but a main challenge in urban ecology is to better understand the factors causing different distributions among species in response to urbanization. Hence, this paper aims to compare the effects of urbanization on woodland plant assemblages in two cities and to describe species responses by using several indicators. The study was carried out in the cities of Angers and Rennes (North-Western France) where 11 isolated woodlands were surveyed along an urban-rural gradient in each city. Abundance data of spontaneous species were collected from 220 quadrats. The effect of land cover (within a 500 m buffer around each woodland) on species assemblages was investigated by Canonical Correspondence Analysis. Buildings and pavement areas were the most significant predictors of species composition, and the effect of location in Angers or Rennes appeared on the second axis. More than 60% of the most frequent plant specieswere indicator of urban or rural location and their preferences were similar in the two cities. These lists of urban and rural indicator species were compared with Ellenberg's indicator values and two other indicators specific to forest environment. The species which grow preferentially in urban woodlands are species which are already known to be associated with recent forests rather than ancient forests; with hedgerows rather than woodlands. The opposite pattern was observed concerning rural species. Moreover, urban indicator species have higher optima for soil pH and soil nitrogen content than rural indicator species. Different characteristics and history of forest habitat—continuity of the forest land cover, linearity of the habitat, change in adjacent land cover and land use—could select the same species, and the responses of the latter might involve different preferences concerning soil alkalinity and nutrient status