Characteristics of intracellular peptidase and proteinase activities from the mycelium of a cord-forming wood decay fungus Serpula lacrymans

Serpula lacrymans is a basidiomycete cord-forming wood decay fungus which reallocates nitrogen within an extensive perennial mycelial system in response to spatial discontinuities in external nutrient supply. Intracellular stored protein is mobilised by conversion to amino acids at nutrient-poor sites within a mycelium or when a whole mycelium is starved. Intracellular peptidase and proteinase activities of the mycelium were investigated with the aim of identifying proteases specifically activated in response to a nitrogen demand. Mycelium for enzyme extraction grown as surface mats in static liquid culture was homogenised, and the extract used in assays for proteinase and peptidase with various synthetic peptide substrates conjugated to 4-nitroaniline. Activities against different substrates were characterised with respect to pH, inhibitor sensitivity, requirements for divalent metal ions, isoelectric point, and by changes in activities in starved mycelium. Four different activities were found, comprising two peptidases one of which had metalloprotease characteristics, a serine-type proteinase, and a proteinase active at pH 2.5 which was not affected by any of the inhibitors tried. Both the latter were most active in starved mycelium. Isoelectric focusing showed peaks with activities corresponding to the serine-type proteinase and one of the manganese-activated peptidases. Possible roles for these enzymes in nitrogen reallocation during mycelial foraging are discussed

Range maps and checklists provide similar estimates of taxonomic and phylogenetic alpha diversity, but less so for beta diversity, of Brazilian Atlantic Forest anurans

AbstractMacroecological and biogeographical studies have assumed that range map data should be used only at coarser grains due to false presences (errors of commission) at small grains. This has been explored using mostly species richness, underrepresenting other potentially informative biodiversity metrics. Here, we evaluated these issues by quantifying the extent to which taxonomic and phylogenetic alpha and beta diversity patterns calculated using anuran range maps at three cell sizes (1×1km, 5×5km, and 10×10km) differ from the patterns calculated based on checklists in 14 protected areas along the southern range of the Brazilian Atlantic Forest. We found that range maps and checklists generated reasonably similar spatial richness patterns in all cell sizes (r≥0.80 in all cases) and slightly weaker, but still correlated alpha phylogenetic diversity patterns (0.78≤r≤0.81). We also found that taxonomic (r≤0.76) and phylogenetic (r≤0.68) beta diversities had lower correlations than alpha spatial patterns. Therefore, range maps have value in documenting alpha biodiversity patterns, as well as beta diversity at more marginal levels, for tropical species at scales relevant to local conservation efforts

Functionalised thermally induced phase separation (TIPS) microparticles enabled for “click” chemistry

Due to their homogeneity, tuneable properties, low cost and ease of manufacture, thermally induced phase separation (TIPS) polymeric microparticles are emerging as an exciting class of injectable device for the treatment of damaged tissue or complex diseases, such as cancer. However, relatively little work has explored enhancing surface functionalisation of this system. Herein, we present the functionalisation of TIPS microparticles with both small molecules and an antibody fragment of Herceptin™, via a heterobifunctional pyridazinedione linker capable of participating in SPAAC “click” chemistry, and compare it to the traditional method of preparing active-targeted microparticle systems, that is, physisorption of antibodies to the microparticle surface. Antigen-binding assays demonstrated that functionalisation of microparticles with Herceptin Fab, via a pyridazinedione linker, provided an enhanced avidity to HER2+ when compared to traditional physisorption methods

Increasing Access to Natural Areas: Connecting Physical and Social Dimensions

Report of the 2015 Berkley Workshop Held at the Asticou Inn, Northeast Harbor, Maine - July 201

BILSAT: Advancing Smallsat Capabilities

Small spacecraft technologies and capabilities are evolving to the point where the BILSAT 120kg spacecraft will this year demonstrate capabilities and performance similar to the 320kg UoSAT-12 mission launched in 1999. Over the past few years, the design of small satellites has evolved from simple curiosities to effective, high performance systems, capable of competing with much bigger and much more expensive spacecraft. Within the framework of an agreement between SSTL and TUBITAK-BILTEN (The Information Technologies and Electronics Research Institute), a non-profit government laboratory located in Ankara, Turkey, a Technology Transfer Program was started in August 2001. This program includes the design, manufacture and launch of one Enhanced SSTL microsatellite platform, one engineering model for use in Turkey and the training of engineers in all aspects of the spacecraft design. Detailed design began using the Enhanced SSTL microsatellite platform as the starting point. The end product that will be launched in the summer of 2003, is the most advanced spacecraft ever designed by SSTL, carrying two advanced payloads developed by TUBITAK-BILTEN. The spacecraft is a highly optimised satellite, with a mass of 120kg and including 14 cameras (in several imager arrangements), a 10m/s class resistojet propulsion system, VHF/UHF and S-band RF systems, tried and tested OBDH units in parallel with newly designed mass data storage and processing units, all this topped by a high performance AODCS subsystem, including two star trackers, GPS receiver (for both orbit and attitude determination), rate gyros, four momentum/reaction wheels, and what will be the first operational use of Control Momentum Gyros on a small spacecraft, to perform high agility manoeuvres. These units will be used to achieve the missions specified for this project, mainly full imaging of Turkey, stereoscopic imaging of selected targets, a Digital Elevation Map of Turkey, and communications. The present paper discusses briefly the technical characteristics of the spacecraft, but focuses on the mission aspects and how the different subsystems (namely the new subsystems and payloads) will be used to accomplish the mission. The operational modes of the spacecraft are discussed and the interaction of the AODCS subsystem with the OBDH and Imaging system is described in detail

Police legitimacy among immigrants in Europe: institutional frames and group position

Research on the antecedents of police legitimacy has begun to stress the relevance of a wide range of factors – beyond performance – in shaping public judgements of police (e.g. Jackson et al 2012; Antrobus et al 2015; Mehozay and Factor 2016; Weitzer and Tuch 2006). The ways in which people experience not just policing and but also their wider social, cultural and economic environment – and the location of both police and policed within structures of power, authority and affect – have important effects on lay judgements of police which, in turn, constitute the empirical legitimacy of this foundational state institution

The in vitro effects of resistin on the innate immune signaling pathway in isolated human subcutaneous adipocytes

Context: Obesity-associated inflammation is a contributory factor in the pathogenesis of type 2 diabetes mellitus (T2DM); the mechanisms underlying the progression to T2DM are unclear. The adipokine resistin has demonstrated pro-inflammatory properties in relation to obesity and T2DM. Objective: To characterize resistin expression in human obesity and address the role of resistin in the innate immune pathway. Furthermore, examine the influence of lipopolysaccharide, recombinant human resistin (rhResistin), insulin and rosiglitazone in human adipocytes. Finally, analyze the effect of rhResistin on the expression of components of the NF-κB pathway and insulin signaling cascade. Methods: Abdominal subcutaneous adipose tissue was obtained from patients undergoing elective liposuction surgery (n = 35, aged: 36-49 yr; BMI: 26.5 ± 5.9 kg/m2). Isolated adipocytes were cultured with rhResistin (10-50 ng/ml). The level of cytokine secretion from isolated adipocytes was examined by ELISA. The effect of rhResistin on protein expression of components of the innate immune pathway was examined by Western blot. Results: In-vitro studies demonstrated that antigenic stimuli increase resistin secretion (P < 0.001) from isolated adipocytes. Pro-inflammatory cytokine levels were increased in response to rhResistin (P < 0.001); this was attenuated by rosiglitazone (P < 0.01). When examining components of the innate immune pathway, rhResistin stimulated Toll-like receptor-2 protein expression. Similarly, mediators of the insulin signaling pathway, phosphospecific JNK1 and JNK2, were upregulated in response to rhResistin. Conclusion: Resistin may participate in more than one mechanism to influence pro-inflammatory cytokine release from human adipocytes; potentially via the integration of NF-κB and JNK signaling pathways

The MINERν\nuA Data Acquisition System and Infrastructure

MINER$\nu$A (Main INjector ExpeRiment $\nu$-A) is a new few-GeV neutrino cross section experiment that began taking data in the FNAL NuMI (Fermi National Accelerator Laboratory Neutrinos at the Main Injector) beam-line in March of 2010. MINER$\nu$A employs a fine-grained scintillator detector capable of complete kinematic characterization of neutrino interactions. This paper describes the MINER$\nu$A data acquisition system (DAQ) including the read-out electronics, software, and computing architecture.Comment: 34 pages, 16 figure

Contribution of Cystine-Glutamate Antiporters to the Psychotomimetic Effects of Phencyclidine

Altered glutamate signaling contributes to a myriad of neural disorders, including schizophrenia. While synaptic levels are intensely studied, nonvesicular release mechanisms, including cystine–glutamate exchange, maintain high steady-state glutamate levels in the extrasynaptic space. The existence of extrasynaptic receptors, including metabotropic group II glutamate receptors (mGluR), pose nonvesicular release mechanisms as unrecognized targets capable of contributing to pathological glutamate signaling. We tested the hypothesis that activation of cystine–glutamate antiporters using the cysteine prodrug N-acetylcysteine would blunt psychotomimetic effects in the rodent phencyclidine (PCP) model of schizophrenia. First, we demonstrate that PCP elevates extracellular glutamate in the prefrontal cortex, an effect that is blocked by N-acetylcysteine pretreatment. To determine the relevance of the above finding, we assessed social interaction and found that N-acetylcysteine reverses social withdrawal produced by repeated PCP. In a separate paradigm, acute PCP resulted in working memory deficits assessed using a discrete trial t-maze task, and this effect was also reversed by N-acetylcysteine pretreatment. The capacity of N-acetylcysteine to restore working memory was blocked by infusion of the cystine–glutamate antiporter inhibitor (S)-4-carboxyphenylglycine into the prefrontal cortex or systemic administration of the group II mGluR antagonist LY341495 indicating that the effects of N-acetylcysteine requires cystine–glutamate exchange and group II mGluR activation. Finally, protein levels from postmortem tissue obtained from schizophrenic patients revealed significant changes in the level of xCT, the active subunit for cystine–glutamate exchange, in the dorsolateral prefrontal cortex. These data advance cystine–glutamate antiporters as novel targets capable of reversing the psychotomimetic effects of PCP