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

Distributions of particulate Heme b in the Atlantic and Southern Oceans-Implications for electron transport in phytoplankton

Concentrations of heme b, the iron-containing component of b-type hemoproteins, ranged from  500). High chl a:heme b ratios resulted from relative decreases in heme b, suggesting proteins such as cytochrome b6f, the core complex of photosystem II, and eukaryotic nitrate reductase were depleted relative to proteins containing chlorophyll such as the eukaryotic light-harvesting antenna. Relative variations in heme b, particulate organic carbon, and chl a can thus be indicative of a physiological response of the phytoplankton community to the prevailing growth conditions, within the context of large-scale changes in phytoplankton community composition

HIPERCAPITALISMO, FLEXIBILIZAÇÃO E METAMORFOSES NO TRABALHO MÉDICO

As transformações aceleradas que se operam no mundo do trabalho no limiar do século XXI vem impactando significativamente as atividades profissionais nas organizações. Nesta entrevista, o Prof. Dr. Josep Blanch aborda o novo paradigma forjado pelo hipercapitalismo que vem reconfigurando o significado do trabalho e a prática da medicina. Ele argumenta que tais transformações são indissociáveis da vivência de conflitos e contradições entre os imperativos funcionais da nova ordem sanitária e os valores, normas, crenças, papéis e padrões morais de comportamento profissional. O Prof. Dr. Blanch, psicólogo, é altamente qualificado para tratar da temática em foco. Ele possui carreira acadêmica sólida, cuja referência central é o trabalho realizado na Universidade Autônoma de Barcelona (UAB/ES), vasta produção bibliográfica acadêmica, experiência internacional relevante com destaque em países da América Latina. Sua expertise profissional também se expressa pelos títulos de Professor Emérito na UAB e o obtido no Ministério da Ciências da Colômbia como Pesquisador Emérito. A entrevista nasceu da Roda de Conversa promovida na disciplina Trabalho e Cognição do Programa de Pós-Graduação em Psicologia Social, do Trabalho e das Organizações (PPG-PSTO) da Universidade de Brasília (UnB). O coletivo de estudantes de pós-graduação (entrevistadores) elaborou questões prévias que foram respondidas durante uma live com quase duas horas de duração

Environmental Metal Pollution Considered as Noise: Effects on the Spatial Distribution of Benthic Foraminifera in two Coastal Marine Areas of Sicily (Southern Italy)

We analyze the spatial distributions of two groups of benthic foraminifera (Adelosina spp. + Quinqueloculina spp. and Elphidium spp.), along Sicilian coast, and their correlation with six different heavy metals, responsible for the pollution. Samples were collected inside the Gulf of Palermo, which has a high level of pollution due to heavy metals, and along the coast of Lampedusa island (Sicily Channel, Southern Mediterranean), which is characterized by unpolluted sea waters. Because of the environmental pollution we find: (i) an anticorrelated spatial behaviour between the two groups of benthic foraminifera analyzed; (ii) an anticorrelated (correlated) spatial behaviour between the first (second) group of benthic foraminifera with metal concentrations; (iii) an almost uncorrelated spatial behaviour between low concentrations of metals and the first group of foraminifera in clean sea water sites. We introduce a two-species model based on the generalized Lotka-Volterra equations in the presence of a multiplicative noise, which models the interaction between species and environmental pollution due to the presence in top-soft sediments of heavy metals. The interaction coefficients between the two species are kept constant with values in the coexistence regime. Using proper values for the initial conditions and the model parameters, we find for the two species a theoretical spatial distribution behaviour in a good agreement with the data obtained from the 63 sites analyzed in our study.Comment: 28 pages, 8 figures, 5 table

Elucidation of the ATP7B N-Domain Mg2+-ATP Coordination Site and Its Allosteric Regulation

The diagnostic of orphan genetic disease is often a puzzling task as less attention is paid to the elucidation of the pathophysiology of these rare disorders at the molecular level. We present here a multidisciplinary approach using molecular modeling tools and surface plasmonic resonance to study the function of the ATP7B protein, which is impaired in the Wilson disease. Experimentally validated in silico models allow the elucidation in the Nucleotide binding domain (N-domain) of the Mg2+-ATP coordination site and answer to the controversial role of the Mg2+ ion in the nucleotide binding process. The analysis of protein motions revealed a substantial effect on a long flexible loop branched to the N-domain protein core. We demonstrated the capacity of the loop to disrupt the interaction between Mg2+-ATP complex and the N-domain and propose a role for this loop in the allosteric regulation of the nucleotide binding process

Minimal Functional Sites Allow a Classification of Zinc Sites in Proteins

Zinc is indispensable to all forms of life as it is an essential component of many different proteins involved in a wide range of biological processes. Not differently from other metals, zinc in proteins can play different roles that depend on the features of the metal-binding site. In this work, we describe zinc sites in proteins with known structure by means of three-dimensional templates that can be automatically extracted from PDB files and consist of the protein structure around the metal, including the zinc ligands and the residues in close spatial proximity to the ligands. This definition is devised to intrinsically capture the features of the local protein environment that can affect metal function, and corresponds to what we call a minimal functional site (MFS). We used MFSs to classify all zinc sites whose structures are available in the PDB and combined this classification with functional annotation as available in the literature. We classified 77% of zinc sites into ten clusters, each grouping zinc sites with structures that are highly similar, and an additional 16% into seven pseudo-clusters, each grouping zinc sites with structures that are only broadly similar. Sites where zinc plays a structural role are predominant in eight clusters and in two pseudo-clusters, while sites where zinc plays a catalytic role are predominant in two clusters and in five pseudo-clusters. We also analyzed the amino acid composition of the coordination sphere of zinc as a function of its role in the protein, highlighting trends and exceptions. In a period when the number of known zinc proteins is expected to grow further with the increasing awareness of the cellular mechanisms of zinc homeostasis, this classification represents a valuable basis for structure-function studies of zinc proteins, with broad applications in biochemistry, molecular pharmacology and de novo protein design

Outer Membrane Vesicles Derived from Escherichia coli Induce Systemic Inflammatory Response Syndrome

Sepsis, characterized by a systemic inflammatory state that is usually related to Gram-negative bacterial infection, is a leading cause of death worldwide. Although the annual incidence of sepsis is still rising, the exact cause of Gram-negative bacteria-associated sepsis is not clear. Outer membrane vesicles (OMVs), constitutively secreted from Gram-negative bacteria, are nano-sized spherical bilayered proteolipids. Using a mouse model, we showed that intraperitoneal injection of OMVs derived from intestinal Escherichia coli induced lethality. Furthermore, OMVs induced host responses which resemble a clinically relevant condition like sepsis that was characterized by piloerection, eye exudates, hypothermia, tachypnea, leukopenia, disseminated intravascular coagulation, dysfunction of the lungs, hypotension, and systemic induction of tumor necrosis factor-α and interleukin-6. Our study revealed a previously unidentified causative microbial signal in the pathogenesis of sepsis, suggesting OMVs as a new therapeutic target to prevent and/or treat severe sepsis caused by Gram-negative bacterial infection

Cellular magnesium acquisition : an anomaly in embryonic cation homeostasis

Author Posting. © The Author(s), 2007. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Experimental and Molecular Pathology 83 (2007): 224-240, doi:10.1016/j.yexmp.2007.03.007.The intracellular dominance of magnesium ion makes clinical assessment difficult despite the critical role of Mg++ in many key functions of cells and enzymes. There is general consensus that serum Mg++ levels are not representative of the growing number of conditions for which magnesium is known to be important. There is no consensus method or sample source for testing for clinical purposes. High intracellular Mg++ in vertebrate embryos results in part from interactions of cations which influence cell membrane transport systems. These are functionally competent from the earliest stages, at least transiently held over from the unfertilized ovum. Kinetic studies with radiotracer cations, osmolar variations, media lacking one or more of the four biological cations, Na+, Mg++, K+, and Ca++, and metabolic poison 0.05 mEq/L NaF, demonstrated: (1) all four cations influence the behavior of the others, and (2) energy is required for uptake and efflux on different time scales, some against gradient. Na+ uptake is energy dependent against an efflux gradient. The rate of K+ loss is equal with or without fluoride, suggesting a lack of an energy requirement at these stages. Ca++ efflux took twice as long in the presence of fluoride, likely due in part to intracellular binding. Mg++ is anomalous in that early teleost vertebrate embryos have an intracellular content exceeding the surrounding sea water, an isolated unaffected yolk compartment, and a clear requirement for energy for both uptake and efflux. The physiological, pathological, and therapeutic roles of magnesium are poorly understood. This will change: (1) when 28Mg is once again generally available at a reasonable cost for both basic research and clinical assessment, and (2) when serum or plasma levels are determined simultaneously with intracellular values, preferably as part of complete four cation profiles. Atomic absorption spectrophotometry, energy-dispersive x-ray analysis, and inductively coupled plasma emission spectroscopy on sublingual mucosal and peripheral blood samples are potential methods of value for coordinated assessments.AEC Grant No. 134