Chemotaxonomic markers in Digitalideae (Plantaginaceae)

In a chemosystematic investigation of Digitalideae (Plantaginaceae), the water-soluble part of extracts of two species of Digitalis, two species of Isoplexis, as well as Erinus alpinus and Lafuentea rotundifolia were studied with regard to their content of main carbohydrates, iridoids and caffeoyl phenylethanoid glycosides (CPGs). Digitalis and Isoplexis contained sorbitol, cornoside and a number of other phenylethanoid glycosides including the new tyrosol beta-D-mannopyranoside, sceptroside but were found to lack iridoid glucosides. Erinus contained mainly glucose, the new 8,9-double bond iridoid, erinoside, and a number of known iridoid glucosides including two esters of 6-rhamnopyranosylcatalpol, as well as the CPG poliumoside. Finally, Lafuentea was characterized by the presence of glucose, aucubin and cryptamygin B but apparently lacked CPGs. The chemosystematic significance of the isolated compounds is discussed.     

Short-Term Effects of Cattle Browsing on Tree Sapling Growth in Mountain Wooded Pastures

Browsing by livestock has been identified as an important factor preventing tree regeneration in wooded pastures. Two field experiments were performed to investigate the effects of cattle browsing on tree sapling growth in a mountain-wooded pasture. Two size classes (ca. 12–17 cm and 41–59 cm) of each of 4 species (Picea abies, Abies alba, Acer pseudoplatanus and Fagus sylvatica) were exposed to zero, low and high grazing intensities. We measured the proportion of saplings browsed and the effect of browsing on growth. A mowing treatment within the zero grazing intensity treatment showed no significant effect on sapling growth. One percent of the large saplings but 25% of the small saplings escaped browsing. Saplings were better protected when surrounded by taller vegetation. The proportion of saplings browsed was not significantly different among species although evergreen tree saplings lost a larger proportion of biomass than deciduous species when browsed. Under grazing, average current-year shoot production and total aboveground biomass of all species were significantly reduced. Browsing effects tended to be smaller at the lower grazing intensity. Because the deciduous species were least reduced in aboveground growth, especially under the low grazing intensity, we conclude that at least in short-term, deciduous species are less affected by cattle browsing.     

Methodological contributions towards LC-MS/MS quantification of free VX in plasma: An innovative approach

A novel analytical method has been developed to detect and quantify VX (O-ethyl S-(2(diisopropylamino) ethyl) (methylphosphonothioate)) in plasma using an LC-MS/MS technique. VX detection and quantification in plasma following percutaneous exposure represent a formidable challenge and it is an important part of the ongoing struggle against chemical warfare agents. Liquid-liquid extraction of VX from plasma was performed and it generated a recovery rate of approximately 65% followed by an LC-MS/MS analysis in a 100% organic phase. An Allure biphenyl column (Restek) was tested with detection limit at 0.5 pg/mL (5 μL injected). Initial application was focused on human skin grafted on nude mice as an experimental model with proper adjustments done for very small quantities of plasma.     

Aryl hydrocarbon receptor is critical for homeostasis of invariant gammadelta T cells in the murine epidermis

An immunoregulatory role of aryl hydrocarbon receptor (AhR) has been shown in conventional αβ and γδ T cells, but its function in skin γδ T cells (dendritic epidermal T cells [DETC]) is unknown. In this study, we demonstrate that DETC express AhR in wild-type mice, and are specifically absent in the epidermis of AhR-deficient mice (AhR-KO). We show that DETC precursors are generated in the thymus and home to the skin. Proliferation of DETC in the skin was impaired in AhR-KO mice, resulting in a >90% loss compared with wild type. Surprisingly, DETC were not replaced by αβ T cells or conventional γδ T cells, suggesting a limited time frame for seeding this niche. We found that DETC from AhR-KO mice failed to express the receptor tyrosine kinase c-Kit, a known growth factor for γδ T cells in the gut. Moreover, we found that c-kit is a direct target of AhR, and propose that AhR-dependent c-Kit expression is potentially involved in DETC homeostasis. DETC are a major source of GM-CSF in the skin. Recently, we had shown that impaired Langerhans cell maturation in AhR-KO is related to low GM-CSF levels. Our findings suggest that the DETCs are necessary for LC maturation, and provide insights into a novel role for AhR in the maintenance of skin-specific γδ T cells, and its consequences for the skin immune network.     

Sunflower resistance to multiple downy mildew pathotypes revealed by recognition of conserved effectors of the oomycete Plasmopara halstedii

Over the last 40 years, new sunflower downy mildew isolates (Plasmopara halstedii) have overcome major gene resistances in sunflower, requiring the identification of additional and possibly more durable broad‐spectrum resistances. Here, 354 RXLR effectors defined in silico from our new genomic data were classified in a network of 40 connected components sharing conserved protein domains. Among 205 RXLR effector genes encoding conserved proteins in 17 P. halstedii pathotypes of varying virulence, we selected 30 effectors that were expressed during plant infection as potentially essential genes to target broad‐spectrum resistance in sunflower. The transient expression of the 30 core effectors in sunflower and in Nicotiana benthamiana leaves revealed a wide diversity of targeted subcellular compartments, including organelles not so far shown to be targeted by oomycete effectors such as chloroplasts and processing bodies. More than half of the 30 core effectors were able to suppress pattern‐triggered immunity in N. benthamiana, and five of these induced hypersensitive responses (HR) in sunflower broad‐spectrum resistant lines. HR triggered by PhRXLRC01 co‐segregated with Pl22 resistance in F3 populations and both traits localized in 1.7 Mb on chromosome 13 of the sunflower genome. Pl22 resistance was physically mapped on the sunflower genome recently sequenced, unlike all the other downy mildew resistances published so far. PhRXLRC01 and Pl22 are proposed as an avirulence/resistance gene couple not previously described in sunflower. Core effector recognition is a successful strategy to accelerate broad‐spectrum resistance gene identification in complex crop genomes such as sunflower.     

A novel strategy for delivering Niemann-Pick type C2 proteins across the blood–brain barrier using the brain endothelial-specific AAV-BR1 virus

Treating central nervous system (CNS) diseases is complicated by the incapability of numerous therapeutics to cross the blood–brain barrier (BBB), mainly composed of brain endothelial cells (BECs). Genetically modifying BECs into protein factories that supply the CNS with recombinant proteins is a promising approach to overcome this hindrance, especially in genetic diseases, like Niemann Pick disease type C2 (NPC2), where both CNS and peripheral cells are affected. Here, we investigated the potential of the BEC-specific adeno-associated viral vector (AAV-BR1) encoding NPC2 for expression and secretion from primary BECs cultured in an in vitro BBB model with mixed glial cells, and in healthy BALB/c mice. Transduced primary BECs had significantly increased NPC2 gene expression and secreted NPC2 after viral transduction, which significantly reversed cholesterol deposition in NPC2 deficient fibroblasts. Mice receiving an intravenous injection with AAV-BR1-NCP2-eGFP were sacrificed 8 weeks later and examined for its biodistribution and transgene expression of eGFP and NPC2. AAV-BR1-NPC2-eGFP was distributed mainly to the brain and lightly to the heart and lung, but did not label other organs including the liver. eGFP expression was primarily found in BECs throughout the brain but occasionally also in neurons suggesting transport of the vector across the BBB, a phenomenon also confirmed in vitro. NPC2 gene expression was up-regulated in the brain, and recombinant NPC2 protein expression was observed in both transduced brain capillaries and neurons. Our findings show that AAV-BR1 transduction of BECs is possible and that it may denote a promising strategy for future treatment of NPC2.

     

Impacts of fever on locust life-history traits: costs or benefits?

Fever, like other mechanisms for defence against pathogens, may have positive and negative consequences for host fitness. In ectotherms, fever can be attained through modified behavioural thermoregulation. Here we examine potential costs of behavioural fever by holding adult, gregarious desert locusts at elevated temperatures simulating a range of fever intensities. We found no effect of fever temperatures on primary fitness correlates of survival and fecundity. However, flight capacity and mate competition were reduced, although there was no relation between time spent at fever temperatures and magnitude of the response. While these effects could indicate a direct cost of fever, they are also consistent with a shift towards the solitaria phase state that, in a field context, could be considered an adaptive life-history response to limit the impact of disease. These conflicting interpretations highlight the importance of considering complex defence mechanisms and trade-offs in an appropriate ecological context.