Visualizing cellular heterogeneity by quantifying the dynamics of MAPK activity in live mammalian cells with synthetic fluorescent biosensors.

Mitogen-Activated Protein Kinases (MAPKs) control a wide array of cellular functions by transducing extracellular information into defined biological responses. In order to understand how these pathways are regulated, dynamic single cell measurements are highly needed. Fluorescence microscopy is well suited to perform these measurements. However, more dynamic and sensitive biosensors that allow the quantification of signaling activity in living mammalian cells are required. We have engineered a synthetic fluorescent substrate for human MAPKs (ERK, JNK and p38) that relocates from the nucleus to the cytoplasm when phosphorylated by the kinases. We demonstrate that this reporter displays an improved response compared to other relocation biosensors. This assay allows to monitor the heterogeneity in the MAPK response in a population of isogenic cells, revealing pulses of ERK activity upon a physiological EGFR stimulation. We show applicability of this approach to the analysis of multiple cancer cell lines and primary cells as well as its application in vivo to developing tumors. Using this ERK biosensor, dynamic single cell measurements with high temporal resolution can be obtained. These MAPK reporters can be widely applied to the analysis of molecular mechanisms of MAPK signaling in healthy and diseased state, in cell culture assays or in vivo

Magnetic switching modes for exchange spring systems ErFe2/YFe2/DyFe2/YFe2 with competing anisotropies

The magnetization reversal processes of ½10nm ErFe2=nYFe2=4nm DyFe2=nYFe2" multilayer films with a (110) growth axis and a variable YFe2 layer thickness n are investigated. The magnetically soft YFe2 compound acts as a separator between the hard rare earth (RE) ErFe2 and DyFe2 compounds, each of them bearing different temperature dependent magnetic anisotropy properties. Magnetic measurements of a system with n ¼ 20nm reveal the existence of three switching modes: an independent switching mode at low temperatures, an ErFe2 spin flop switching mode at medium high temperatures, and an YFe2 dominated switching mode at high temperatures. The measurements are in qualitative agreement with the findings of micromagnetic simulations which are used to illustrate the switching modes. Further simulations for a varied YFe2 layer thickness n ranging from 2 to 40nm are carried out. Quantitative criteria are defined to classify the reversal behavior, and the resultant switching modes are laid out in a map with regard to n and the temperature T. A new coupled switching mode emerges above a threshold temperature for samples with thin YFe2 separation layers as a consequence of the exchange coupling between the magnetically hard ErFe2 and DyFe2 layers. It reflects the increasing competition of the two conflicting anisotropies to dominate the magnetic switching states of both RE compounds under decreasing n

Saturnus: Um Simulador Discreto Baseado em Eventos para Sistemas de Arquivos Paralelos

The focus of this work is to study parallel file systems, their elements and present Saturnus, a simulator under development for systems like these. The proposal is to use it to extract data and generate information, focusing on load balancing through the studied environment, showing experiments and possible uses for the tool. (O foco desse trabalho e ́ estudar sistemas de arquivos paralelos, seus elementos e apresentar o Saturnus, um simulador em desenvolvimento para sistemas como esses. A proposta consiste em utilizar o mesmo para extrair dados e gerar informacoes, com foco no balanceamento de carga atrave ́s do ambiente estudado, mostrando experimentos e poss ́ıveis usos para a ferramenta.

Mucoadhesive budesonide formulation for the treatment of eosinophilic esophagitis

Eosinophilic esophagitis (EE) is a chronic immune/antigen-mediated esophageal inflammatory disease for which off-label topical corticosteroids (e.g., budesonide) are widely used in clinic. In general, thickening excipients are mixed with industrial products to improve the residence time of the drug on the esophageal mucosa. The compounding procedures are empirical and the composition is not supported by real physicochemical and technological characterization. The current study aimed to propose a standardized budesonide oral formulation intended to improve the resistance time of the drug on the esophageal mucosa for EE treatment. Different placebo and drug-loaded (0.025% w/w) formulations were prepared by changing the percentage of xanthan gum alone or in ratio 1:1 with guar gum. Both excipients were added in the composition for their mucoadhesive properties. The formulative space was rationalized based on the drug physicochemical stability and the main critical quality attributes of the formulation, e.g., rheological properties, syringeability, mucoadhesiveness and in vitro penetration of budesonide in porcine esophageal tissue. The obtained results demonstrated that gums allowed a prolonged residence time. However, the concentration of the mucoadhesive polymer has to be rationalized appropriately to permit the syringeability of the formulation and, therefore, easy dosing by the patient/caregiver

A connexin/ifi30 pathway bridges HSCs with their niche to dampen oxidative stress

Reactive oxygen species (ROS) represent a by-product of metabolism and their excess is toxic for hematopoietic stem and progenitor cells (HSPCs). During embryogenesis, a small number of HSPCs are produced from the hemogenic endothelium, before they colonize a transient organ where they expand, for example the fetal liver in mammals. In this study, we use zebrafish to understand the molecular mechanisms that are important in the caudal hematopoietic tissue (equivalent to the mammalian fetal liver) to promote HSPC expansion. High levels of ROS are deleterious for HSPCs in this niche, however this is rescued by addition of antioxidants. We show that Cx41.8 is important to lower ROS levels in HSPCs. We also demonstrate a new role for ifi30, known to be involved in the immune response. In the hematopoietic niche, Ifi30 can recycle oxidized glutathione to allow HSPCs to dampen their levels of ROS, a role that could be conserved in human fetal liver

Role of magnetic dopants in the phase diagram of Sm1111 pnictides: The Mn case

The deliberate insertion of magnetic Mn dopants in the Fe sites of the optimally-doped SmFeAsO0.88-F0.12 iron-based superconductor can modify in a controlled way its electronic properties. The resulting phase diagram was investigated across a wide range of manganese contents (x) by means of muon-spin spectroscopy (muSR), both in zero- and in transverse fields, respectively, to probe the magnetic and the superconducting order. The pure superconducting phase (at x < 0.03) is replaced by a crossover region at intermediate Mn values (0.03 =< x < 0.08), where superconductivity coexists with static magnetic order. After completely suppressing superconductivity for x = 0.08, a further increase in Mn content reinforces the natural tendency towards antiferromagnetic correlations among the magnetic Mn ions. The sharp drop of Tc and the induced magnetic order in the presence of magnetic disorder/dopants, such as Mn, are both consistent with a recent theoretical model of unconventional superconductors [M. Gastiasoro et al., ArXiv 1606.09495], which includes correlation-enhanced RKKY-couplings between the impurity moments.Comment: 10 pages, 13 figures. Accepted on Physical Review

Effect of Feed Restriction on the Behaviour and Welfare of Broiler Chickens

Under intensive rearing conditions, the welfare of broiler chickens may be at risk depending on genotype and sex, due to their different growth rates. The practice of quantitative feed restriction may also impact on welfare. This study aimed to evaluate behaviour and corticosterone content in plasma and faeces at different ages using 896 one-day-old chicks housed in 32 pens, allocated to 8 groups, i.e., 2 genotypes (standard vs. high breast yield) 7 2 sex 7 2 feeding plans (ad libitum vs. restricted, AL vs. FR). The feeding system affected the percentage of standing (9.84% vs. 11.7% in AL vs. FR; p 64 0.001), feeding (7.51% vs. 8.17%; p 64 0.01) and sitting/lying (67.0% vs. 64.1%; p 64 0.001), and the faeces corticosterone content (12.2 vs. 13.6 ng/g in AL vs. FR; p 64 0.10). Sex affected the percentage of pecking other chickens, standing and comfort behaviours. Changes in behaviour were recorded between high and standard breast yield genotypes with faeces corticosterone which tended to be higher in the former (p 64 0.10). Significant interactions between the main factors and age were observed. Major changes in behaviour were due to feed restriction, which stimulated activity during restriction

Release of Palladium from Biomechanical Prostheses in Body Fluids Can Induce or Support PD-Specific IFNγ T Cell Responses and the Clinical Setting of a Palladium Hypersensitivity:

The increased use of Palladium (Pd) for biomedical applications, which has more than doubled in the last ten years, appears to be associated with an increased frequency of adverse reactions to Pd. The aim of this study is to investigate the relationship between the implant of a biomechanical apparatus containing Pd and the setting of a hypersensitivity to Pd by determining the levels of the metal released in biological fluids, assessing the effects of Pd on peripheral blood mononuclear cell (PBMC) cytokine production and exploring the clinical setting of skin sensitization. Of a total of 3,093 subjects examined in 2006, sensitization to Pd alone or in association with nickel (Ni) was observed in 1.6% and 13.03% of the individuals, respectively. Of these, a group of six subjects positive to Pd and negative to Ni at patch testing were selected on the basis of the oral clinical symptoms in order to measure both the levels of Pd in biological fluids and the degradation of the dental prostheses. Specific Pd measurements were carried out on salivary fluid, urine and serum samples by High Resolution Inductively Coupled Plasma-Mass Spectrometry. In addition, the degradation of the dental prostheses was assessed by both a "leaching test" and an analysis of the micro morphology of orthodontic prostheses. The induction of IFN-γ production by Pd was assessed in PBMC by the ELISpot assay. Skin sensitization to Pd was evaluated by patch testing and clinical examination. Ten healthy subjects were comparatively tested as controls. We found a specific induction of an IFN-γ response by Pd in PBMC collected from all the subjects positive to Pd at patch testing. On the contrary, control subjects did not show any response to Pd as assessed by IFN-γ ELISpot assay or by skin testing. Remarkably, the levels of Pd in all biological samples (saliva, sera, urine) were significantly higher in Pd-sensitized patients than in those collected from controls, reaching the highest concentrations in the urine. The leaching studies gave additional evidence that the dental appliances can release measurable levels of Pd in saliva. Oral clinical symptoms in patients with Pd dental prostheses were associated with measurable levels of Pd in the biological fluids, the induction of Pd-specific IFN-γ responses in PBMC and the clinical evidence of skin sensitization to Pd. These data suggest that dental appliances may represent an active source of Pd in the body, and this, in turn, can favour the clinical setting of a hypersensitivity to this metal

Comunidade de pequenos mamíferos de paisagens nativas e pastagens cultivadas no Pantanal da Nhecolândia.

O Pantanal possui uma grande diversidade de habitats, compondo uma paisagem complexa que tem sido conservada pela pecuária tradicional. Mais recentemente, a implantação de pastagem cultivada, visando ao aumento da produtividade e intensificando o sistema de produção vem alterando a paisagem pantaneira. Os resultados dessa alteração é a simplificação da composição e da complexidade da paisagem, colocando em risco a biodiversidade regional. O objetivo deste estudo foi avaliar alterações na comunidade de pequenos mamíferos em diferentes fitofisionomias, inclusive área alterada, em duas Fazendas no Pantanal da Nhecolândia. The Pantanal presents a great habitat diversity composing a complex landscape which has been conserved by the traditional cattle ranching. Recently, the cultivation of pastures aiming the increase of the productivity and intensification of the production system has been altering the landscape in the Pantanal. The results of this alteration é the simplification of the landscape composition and complexity, placing at risk the regional biodiversity. The objective of this study was to evaluate the changes in the small mammal communities at different phytophysiognomies, including modified areas, in two ranches at the Nhecolândia region of the Pantanal