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Chronic use of statins may have anti-inflammatory action, promoting immunomodulation and survival in patients with sepsis. This study aimed to analyze the effects of pretreatment with simvastatin in lethal sepsis induced by cecal ligation and puncture (CLP). Male Swiss mice received prophylactic treatment with simvastatin or pyrogen-free water orally in a single daily dose for 30 days. After this period, the CLP was performed. Naïve and Sham groups were performed as non-infected controls. Animal survival was monitored for 60 h after the CLP. Half of mice were euthanized after 12 h to analyze colony-forming units (CFUs); hematological parameters; production of IL-10, IL-12, IL-6, TNF-α, IFN-γ, and MCP-1; cell counts on peritoneum, bronchoalveolar lavage (BAL), bone marrow, spleen, and mesenteric lymph node; immunephenotyping of T cells and antigen presenting cells and production of hydrogen peroxide (H2O2). Simvastatin induced an increase in survival and a decrease in the CFU count on peritoneum and on BAL cells number, especially lymphocytes. There was an increase in the platelets and lymphocytes number in the Simvastatin group when compared to the CLP group. Simvastatin induced a greater activation and proliferation of CD4+ T cells, as well as an increase in IL-6 and MCP-1 production, in chemotaxis to the peritoneum and in H2O2 secretion at this site. These data suggest that simvastatin has an impact on the survival of animals, as well as immunomodulatory effects in sepsis induced by CLP in mice.

Bufadienolides are cardiotonic steroids (CTS) identified in mammals. Besides Na+/K+-ATPase inhibition, they activate signal transduction via protein&ndash, protein interactions. Diversity of endogenous bufadienolides and mechanisms of action may indicate the presence of functional selectivity and unique cellular outcomes. We evaluated whether the bufadienolides telocinobufagin and marinobufagin induce changes in proliferation or viability of pig kidney (LLC-PK1) cells and the mechanisms involved in these changes. In some experiments, ouabain was used as a positive control. CTS exhibited an inhibitory IC50 of 0.20 (telocinobufagin), 0.14 (ouabain), and 3.40 &mu, M (marinobufagin) for pig kidney Na+/K+-ATPase activity and concentrations that barely inhibited it were tested in LLC-PK1 cells. CTS induced rapid ERK1/2 phosphorylation, but corresponding proliferative response was observed for marinobufagin and ouabain instead of telocinobufagin. Telocinobufagin increased Bax:Bcl-2 expression ratio, sub-G0 cell cycle phase and pyknotic nuclei, indicating apoptosis. Src and MEK1/2 inhibitors blunted marinobufagin but not telocinobufagin effect, which was also not mediated by p38, JNK1/2, and PI3K. However, BIO, a GSK-3&beta, inhibitor, reduced proliferation and, as telocinobufagin, phosphorylated GSK-3&beta, at inhibitory Ser9. Combination of both drugs resulted in synergistic antiproliferative effect. Wnt reporter activity assay showed that telocinobufagin impaired Wnt/&beta, catenin pathway by acting upstream to &beta, catenin stabilization. Our findings support that mammalian endogenous bufadienolides may exhibit functional selectivity.

Na implementacao de planos de gerenciamento hidrico, se faz necessario compreender o comportamento dos agentes consumidores deagua diante de decisoes, planos e projetos de gerenciamento do sistema hidrico. Essa integracao possibilita tomadas de decisao por parte dos gestores em concordância com a realidade do sistema hidrico. No presente trabalho se apresenta a ferramenta Hydric-Agent desenvolvida em JADEX que permite a simulacao de consumidores deagua sob diferentes cenarios de gestao hidrica, fornecendo ao usuario da plataforma uma visao do cooperativismo do agente consumidor deagua e o grau de adesaoa acao de gestao estabelecida.

Anacardic acid, a phenolic compound, represents 90% of cashew nut shell liquid, which is a byproduct from the industrial processing of cashew nuts. This study aimed to add calcium anacardate (CA) to broilers' diets as a source of anacardic acid, to evaluate its antioxidant effect in breast meat and in processed meat products (sausages). For this purpose, birds were fed according to the following treatments: diet without antioxidant and diets containing 2.5, 5.0, 7.5, or 10.0 g kgFor breast meat, a 2.5 g kgCalcium anacardate is a potential natural antioxidant for breast meat and sausages in storage when added to broilers' diets. © 2018 Society of Chemical Industry.

The yellow fever virus (YFV) epidemic that began in Dec 2016 in Brazil is the largest in decades. The recent discovery of YFV in BrazilianAedes sp.vectors highlights the urgent need to monitor the risk of re-establishment of domestic YFV transmission in the Americas. We use a suite of epidemiological, spatial and genomic approaches to characterize YFV transmission. We show that the age- and sex-distribution of human cases in Brazil is characteristic of sylvatic transmission. Analysis of YFV cases combined with genomes generated locally using a new protocol reveals an early phase of sylvatic YFV transmission restricted to Minas Gerais, followed in late 2016 by a rise in viral spillover to humans, and the southwards spatial expansion of the epidemic towards previously YFV-free areas. Our results establish a framework for monitoring YFV transmission in real-time, contributing to the global strategy of eliminating future yellow fever epidemics.

Leptospires are highly motile spirochetes equipped with strategies for efficient invasion and dissemination within the host. Our group previously demonstrated that pathogenic leptospires secrete proteases capable of cleaving and inactivating key molecules of the complement system, allowing these bacteria to circumvent host's innate immune defense mechanisms. Given the successful dissemination of leptospires during infection, we wondered if such proteases would target a broader range of host molecules. In the present study, the proteolytic activity of secreted leptospiral proteases against a panel of extracellular matrix (ECM) and plasma proteins was assessed. The culture supernatant of the virulent L. interrogans serovar Kennewicki strain Fromm (LPF) degraded human fibrinogen, plasma fibronectin, gelatin, and the proteoglycans decorin, biglycan, and lumican. Interestingly, human plasminogen was not cleaved by proteases present in the supernatants. Proteolytic activity was inhibited by 1,10-phenanthroline, suggesting the participation of metalloproteases. Moreover, production of proteases might be an important virulence determinant since culture-attenuated or saprophytic Leptospira did not display proteolytic activity against ECM or plasma components. Exoproteomic analysis allowed the identification of three metalloproteases that could be involved in the degradation of host components. The ability to cleave conjunctive tissue molecules and coagulation cascade proteins may certainly contribute to invasion and tissue destruction observed upon infection with Leptospira.

This paper presents an overview of uncertainty assessment in agent-based simulations, mainly related to land use and cover change. Almost every multiagent-based simulation review has expressed the need for statistical methods to evaluate the certainty of the results. Yet these problems continue to be underestimated and often neglected. This work aims to review how uncertainty is being portrayed in agent-based simulation and to perform an exploratory study to use statistical methods to estimate uncertainty. MASE, a Multi-Agent System for Environmental simulation, is the system under study. We first identified the most sensitive parameters using Morris One-at-a-Time sensitivity analysis. The efforts to assess agent-based simulation through statistical methods are paramount to corroborate and improve the level of confidence of the research that has been made in land use simulation.

The incidence of infections caused by rapidly growing mycobacteria (RGM), especially Mycobacterium abscessus subsp. massiliense (Mabs), is increasing worldwide. Severe infections are associated with abscess formation and strong inflammatory response. This study evaluated the antimicrobial and anti-inflammatory activities of a hydroalcoholic extract (BoHE) and ethyl acetate fraction (BoEA) of Bixa orellana leaves. Antimicrobial activity was evaluated by broth microdilution to determine the minimum inhibitory (MIC) and the minimum bactericidal (MBC) concentrations. Cytotoxicity was evaluated using erythrocytes and RAW 264.7 cells. Nitric oxide (NO) was assayed in stimulated RAW 264.7 cells, and inflammatory cell migration and acute toxicity were evaluated in a Mabs-induced peritonitis mouse model. The compounds present in BoEA were identified by high performance liquid chromatography and mass spectrometry (HPLC-MS). The MIC and MBC values were 2.34 mg/mL and 37.5 mg/mL for BoHE and 0.39 mg/mL and 6.25 mg/mL for BoEA. The extracts did not induce significant toxicity in erythrocytes and RAW 264.7 cells. High levels of NO induced by Mabs were decreased by treatment with both extracts. The anti-inflammatory activity was confirmed in vivo by significant reduction of the cell migration to the peritoneum following BoHE and BoEA pretreatment. Animals treated with BoHE or BoEA did not show signs of acute toxicity in stomach, liver, and kidney. The chemical characterization of BoEA (the most active extract) revealed that kaempferol-3-O-coumaroyl glucose is its major component. The extract of B. orellana may be effective for treating infections caused by Mabs.

Human adipose-derived stromal cells (hADSC) are a heterogeneous cell population that contains adult multipotent stem cells. Although it is well established that hADSC have skeletal potential in vivo in adult organisms, in vitro assays suggest further differentiation capacity, such as into glia. Thus, we propose that grafting hADSC into the embryo can provide them with a much more instructive microenvironment, allowing the human cells to adopt diverse fates or niches. Here, hADSC spheroids were grafted into either the presumptive presomitic mesoderm or the first branchial arch (BA1) regions of chick embryos. Cells were identified without previous manipulations via human-specific Alu probes, which allows efficient long-term tracing of heterogeneous primary cultures. When grafted into the trunk, in contrast to previous studies, hADSC were not found in chondrogenic or osteogenic territories up to E8. Surprisingly, 82.5% of the hADSC were associated with HNK1+ tissues, such as peripheral nerves. Human skin fibroblasts showed a smaller tropism for nerves. In line with other studies, hADSC also adopted perivascular locations. When grafted into the presumptive BA1, 74.6% of the cells were in the outflow tract, the final goal of cardiac neural crest cells, and were also associated with peripheral nerves. This is the first study showing that hADSC could adopt a perineural niche in vivo and were able to recognize cues for neural crest cell migration of the host. Therefore, we propose that xenografts of human cells into chick embryos can reveal novel behaviors of heterogeneous cell populations, such as response to migration cues.

Summary Secreted Wnt morphogens are essential for embryogenesis and homeostasis and require a lipid/palmitoleoylate modification for receptor binding and activity. Notum is a secreted Wnt antagonist that belongs to the α/β hydrolase superfamily, but its mechanism of action and roles in vertebrate embryogenesis are not fully understood. Here, we report that Notum hydrolyzes the Wnt palmitoleoylate adduct extracellularly, resulting in inactivated Wnt proteins that form oxidized oligomers incapable of receptor binding. Thus, Notum is a Wnt deacylase, and palmitoleoylation is obligatory for the Wnt structure that maintains its active monomeric conformation. Notum is expressed in naive ectoderm and neural plate in Xenopus and is required for neural and head induction. These findings suggest that Notum is a prerequisite for the "default" neural fate and that distinct mechanisms of Wnt inactivation by the Tiki protease in the Organizer and the Notum deacylase in presumptive neuroectoderm orchestrate vertebrate brain development.

This experiment was designed to evaluate the effects of the inclusion of detoxified castor bean meal (CM) content on the chemical-bromatological composition, as well as the fermentation characteristics and dry matter losses of sugarcane silage. The treatments consisted of four levels (0, 7, 14 and 21% natural matter) of addition of castor bean meal. The design was completely randomized, with five replicates. The material was ensiled in PVC silos of 50-cm in height and 10-cm in diameter, which were opened after 60 days. The density of fodder maintained in the silos was equal to 750kg of natural matter/m3. In order to quantify the gas and effluent losses, the silos were weighed at the beginning and at the end of the experiment. The addition of castor bean meal has increased the levels of DM and crude protein and reduced the levels of neutral detergent fiber, N-NH3, ethanol, and gas and effluent losses from silages, but did not affect pH values. During ensiling, alcoholic fermentation was controlled with the inclusion of the additive. O experimento foi desenvolvido para avaliar os efeitos da inclusão de níveis de farelo de mamona destoxificado sobre a composição químico-bromatológica, as características fermentativas e as perdas de matéria seca da silagem de cana-de-açúcar. Os tratamentos consistiam em quatro níveis (0, 7, 14 e 21% da matéria natural) de inclusão do farelo de mamona. O delineamento foi inteiramente ao acaso, com cinco repetições. Foram usados silos de PVC, com 50cm de altura e 10cm de diâmetro, para a produção das silagens, que foram abertos após 60 dias. A densidade de forragem nos silos foi correspondente a 750kg de matéria natural/m3. Os silos foram pesados no início e ao final do período experimental para quantificar as perdas por gases e efluentes. A inclusão de farelo de mamona elevou os teores de MS e proteína bruta e reduziu os teores de fibra em detergente neutro, N-NH3, etanol e as perdas por gases e efluentes das silagens, não ocasionando efeito nos valores de pH. A fermentação alcoólica durante a ensilagem foi controlada com a inclusão do aditivo.

Myc proto-oncogenes regulate diverse cellular processes during development, but their roles during morphogenesis of specific tissues are not fully understood. We found that c-myc regulates cell proliferation in mouse lens development and previous genome-wide studies suggested functional roles for N-myc in developing lens. Here, we examined the role of N-myc in mouse lens development. Genetic inactivation of N-myc in the surface ectoderm or lens vesicle impaired eye and lens growth, while "late" inactivation in lens fibers had no effect. Unexpectedly, defective growth of N-myc--deficient lenses was not associated with alterations in lens progenitor cell proliferation or survival. Notably, N-myc-deficient lens exhibited a delay in degradation of DNA in terminally differentiating lens fiber cells. RNA-sequencing analysis of N-myc--deficient lenses identified a cohort of down-regulated genes associated with fiber cell differentiation that included DNaseIIβ. Further, an integrated analysis of differentially expressed genes in N-myc-deficient lens using normal lens expression patterns of iSyTE, N-myc-binding motif analysis and molecular interaction data from the String database led to the derivation of an N-myc-based gene regulatory network in the lens. Finally, analysis of N-myc and c-myc double-deficient lens demonstrated that these Myc genes cooperate to drive lens growth prior to lens vesicle stage. Together, these findings provide evidence for exclusive and cooperative functions of Myc transcription factors in mouse lens development and identify novel mechanisms by which N-myc regulates cell differentiation during eye morphogenesis.

Chenopodium ambrosioides L. (Amaranthaceae) is often used in different kinds of vegetal preparations for medicinal purposes in many clinical situations. Some studies have demonstrated its anti-inflammatory and immunomodulatory properties. The aim of this work was to investigate the effect of prophylactic treatment with the hydroalcoholic extract of C. ambrosioides (HCE) and its hexanic fraction (HEX) on the control of bacterial growth, the activation of phagocytes and the control of the systemic inflammatory response in a sepsis experimental model. Animals were divided into three groups (n = 5/group): Control, which received only NaCl 0.9% solution; HCE, which received the crude extract; and HEX, which received the hexane fraction of the extract. The animals received saline, HCE or HEX (5 mg/kg), subcutaneously (SC), 6 h before cecal ligation and puncture (CLP). Twelve hours after the CLP, the blood was collected to measure the serum cytokines, and the animals were killed for the evaluation of colony-forming units (CFUs), cellular influx and activation of phagocytes in the peritoneal cavity, measured by the secretion of hydrogen peroxide and nitric oxide production. The results showed that only HEX treatment inhibited bacterial growth in the peritoneum and inflammatory cellular influx, especially influx of macrophages and neutrophils. However, HCE and HEX treatments increased ex vivo hydrogen peroxide secretion and nitric oxide production by phagocytes and decreased the pro-inflammatory cytokines in the serum, indicating a systemic anti-inflammatory effect of both. In conclusion, C. ambrosioides treatment decreases bacterial growth likely by activation of phagocytes and, in parallel, ameliorates the general state of mice by reducing the systemic inflammatory response usually observed in sepsis.

This chapter presents research carried out under the MASE project, including the definition of a conceptual model to characterize the behavior of individuals that interact in the dynamics of land-use and cover change. A computational tool for analyzing environmental scenarios of land change was developed, called MASE - Multi-Agent System for Environmental Simulation. MASE enables agent-based simulation scenarios and integrates the influence of socio-economic and political dynamics through the interaction of agents with rules of land-use and planning policies and the environmental physical and spatial variables. MASE simulator was extended to implement the Belief-Desire-Intention (BDI) model, called MASE-BDI. MASE and MASE-BDI are discussed including the conceptual model complexity and statistical techniques of map comparison to land change models. Two real cases of the Brazilian Cerrado validate quantitative and qualitative aspects of MASE and MASE-BDI simulators. Finally, the authors present some auto-tuning aspects of adjusting simulation parameters of MASE-BDI.

It is now well documented that natural products have played an important role in anticancer therapy. Many studies focus on the ability of these natural compounds to modulate tumor-related signaling pathways and the relationship of these properties to an anticancer effect. According to the World Health Organization (WHO), colorectal cancer (CRC) is the third most common cancer and the fourth leading cause of cancer death among men and women. Therefore, finding strategies to fight against CRC is an emergent health problem. CRC has a strong association with deregulation of Wnt/β-catenin signaling pathway. As some types of natural compounds are capable of modulating the Wnt/β-catenin signaling, one important question is whether they could counteract CRC. In this review, we discuss the role of flavonoids, a class of natural compounds, on Wnt/β-catenin regulation and its possible potential for therapeutic usage on colorectal cancer.

The objective was to evaluate the influence of the handling of three milkers on the behavior of dairy cows, milking management and animal production. Direct observations were made, continuous and with visual sampling of the milkers behavior and of the cows. The cows participated of the three treatments (milkers), constituting the cross-over experimental design. In the comparative analysis of the human and animal behavior the nonparametric chi-square (c2) test was used. For data on milk production, flight distance and milking time it was applied the variance analysis (ANOVA). In the economic evaluation it was used a descriptive analysis of the production results. It was concluded that the type of handling of the milker influenced milk production, milking time, flight distance and the reaction of the animals.

Tiki1 is a Wnt protease and antagonist specifically expressed in the Spemann-Mangold Organizer and is required for head formation in Xenopus embryos. Here we report neighbor-joining phylogenetic analysis of vertebrate Tiki genes and their mRNA expression patterns in chick, mouse, and rabbit embryos. Tiki1 and Tiki2 orthologues are highly conserved, and exhibit similar but also different developmental expression patterns among the vertebrate/mammalian species analyzed. The Tiki1 gene is noticeably absent in the rodent lineage, but is present in lagomorphs and all other vertebrate/mammalian species examined. Expression in Hensen's node, the equivalent of the Xenopus Organizer, was observed for Chick Tiki2 and Rabbit Tiki1 and Tiki2. Mouse Tiki2 was detected at low levels at gastrulation and head fold stages, but not in the node. Mouse Tiki2 and chick Tiki1 display similar expression in the dorsal spinal cord. Chick Tiki1 expression was also detected in the surface ectoderm and maxillary bud, while chick Tiki2 was found in the anterior intestinal portal, head mesenchyme and primitive atrium. Our expression analyses provide evidence that Tiki1 and Tiki2 are evolutionarily conserved among vertebrate species and their expression in the Organizer and other regions suggests contributions of these Wnt inhibitors to embryonic patterning, as well as organogenesis. Our analyses further reveal mis-regulation of TIKI1 and TIKI2 in human cancer and diseases.

We investigated the short and long term aging of asphalt cement (AC) with different AFM techniques (topography, phase and friction imaging and nano-indentation experiments). The aging process induces a growth and nucleation of the asphaltene micelles with a concomitant reduction of the maltene phase, whereas the short term aging induces the formation of fractal-like micellar structures. The friction investigation shows that the aging processes reduce the binder friction coefficient by 50%, and this reduction occur predominantly during the short term aging, while the growth of the micelles occur predominantly during the long term aging. The micro-indentation experiments revealed that the aging processes cause a stiffening of the AC film (half-order of magnitude for short term aging, and one order of magnitude for long term aging). The aging process also increased the apparent viscosity of the AC films by half-order of magnitude.

Background: Cuba is a unique country, and despite limited economic development, has an excellent health system. However, the prevalence of asthma symptoms in children in Havana, Cuba, is unusually high. Aim: As early life exposures are critical to the aetiology of asthma, we have studied environmental influences on the risk of wheezing in Cuban infants. Design: Cross-sectional study. Methods: A random sample of 2032 children aged 12–15 months living in Havana was selected for inclusion in the cohort. Data were collected using questionnaires administered by researchers. Results: Of 2032 infants invited to participate, 1956 (96%) infants provided data. The prevalence of any wheeze was 45%, severe wheeze requiring use of emergency services was 30% and recurrent wheeze on three or more occasions was 20%. The largest adjusted risk factors for any wheeze were presence of eczema [odds ratio (OR) 2.09; 95% confidence interval (CI) 1.48–2.94], family history of asthma (OR 2.05; 95% CI 1.60–2.62), poor ventilation in the house (OR 1.99; 95% CI 1.48–2.67), attendance at nursery (OR 1.78; 95% CI 1.24–2.57), male sex (OR1.52; 95% CI 1.19–1.96) and the number of smokers in the house (P < 0.03 for trend), OR 1.64 (95% CI 1.17–2.31) for three or more smokers in the house compared to no smokers in the household. Conclusion: We have identified several risk factors for any wheeze in young infants living in modern day Cuba. As the prevalence of smoking in the house is high (51%), intervention studies are required to determine effective strategies to improve infant health.

In 1919, during colonization of the West Region of São Paulo State, Brazil, the Ogassawara family built a cemetery and a school with donations received from the newspaper Osaka Mainichi Shimbum, in Osaka, Japan. The cemetery was closed by President Getúlio Vargas in 1942, during the Second World War. The architecture of the Japanese cemetery is a unique feature in Latin America. Even considering its historical and cultural relevance, there is a lack of geometric documentation about the location and features of the tombs and other buildings within the cemetery. As an alternative to provide detailed and fast georeferenced information about the area, it is proposed to use near vertical panoramic images taken with a digital camera with fisheye lens as the primary data followed by bundle adjustment and photogrammetric restitution. The aim of this paper is to present a feasibility study on the proposed technique with the assessment of the results with a strip of five panoramic images, taken over some graves in the Japanese cemetery. The results showed that a plant in a scale of 1 : 200 can be produced with photogrammetric restitution at a very low cost, when compared to topographic surveying or laser scanning. The paper will address the main advantages of this technique as well as its drawbacks, with quantitative analysis of the results achieved in this experiment.

O objetivo deste estudo foi avaliar a influência do manejo de 3 ordenhadores sobre o compor-tamento de vacas leiteiras, manejo de ordenha e a produção dos animais, associando a distância de fuga e retorno econômico do manejo. Foram realizadas observações diretas, contínuas e com amostragem visual do com-portamento dos ordenhadores e das vacas. As vacas participaram dos três tratamentos (or-denhadores), constituindo o delineamento experimental cross-over. Na análise comparativa do comportamento humano e animal foi utilizado o teste não paramétrico Qui-quadrado (c2). Para os dados de produção de leite, distância de fuga e tempo de ordenha aplicou-se a análise de variância (ANOVA). Na avaliação econômica utilizou-se uma análise descritiva dos resultados de produção. Concluiu-se que o tipo de manejo do ordenhador influencia as características produtivas e comportamentais de vacas em lactação.

Nanometer thick films of sputtered and evaporated Cu were deposited on the surfaces of the fluoropolymers poly(tetrafluoroethylene) (PTFE), poly(tetrafluoroethylene-co-hexafluoropropylene) (FEP) and poly(tetrafluoroethylene-co-perfluoropropyl vinyl ether) (PFA) and studied by both angle-resolved XPS at takeoff angles of 10°, 45° and 80° and in situ argon ion etching. Higher yields of the fluoride ion to fluoropolymer ratio were detected for sputtered than evaporated Cu. PFA and FEP show enhanced interaction with sputtered Cu to produce fluoride ions relative to the more polycrystalline PTFE. At intermediate depths (takeoff angle of 45°), PFA and FEP exhibit the strongest fluoride F 1s signals compared with the fluoropolymer peaks. The amount of fluoride ion detected reaches a maximum after brief Ar ion etching and then decreases with prolonged etching. Compared with untreated fluoropolymers, improved adhesion of evaporated Cu was observed on the fluoropolymer surfaces that were argon ion etched to expose fluoride ions. Copyright © 2013 John Wiley & Sons, Ltd.

Hospital-acquired infections caused by antibiotic-resistant bacteria threaten the lives of many citizens all over the world. Discovery of new agents to hinder bacterial development would have a significant impact on the treatment of infections. Here, the purification and characterization of Rc-2S-Alb, a protein that belongs to the 2S albumin family, from Ricinus communis seed cake, are reported. Rc-2S-Alb was purified after protein extraction with Tris-HCl buffer, pH 7.5, fractionation by ammonium sulfate (50-75%), and chromatography on Phenyl-Sepharose and DEAE-Sepharose. Rc-2S-Alb, a 75 kDa peptide, displays trypsin inhibitory activity and has high in vitro antibacterial activity against Bacillus subtilis, Klebsiella pneumonia, and Pseudomonas aeruginosa, which are important human pathogenic bacteria. Atomic force microscopy studies indicated that Rc-2S-Alb disrupts the bacterial membrane with loss of the cytoplasm content and ultimately bacterial death. Therefore, Rc-2S-Alb is a powerful candidate for the development of an alternative drug that may help reduce hospital-acquired infections.

Enteroaggregative Escherichia coli (EAEC) is an agent of acute and persistent diarrhea worldwide, categorized in typical or atypical subgroups. Some EAEC virulence factors are members of the serine protease autotransporters of Enterobacteriaceae (SPATE). The presence of SPATE-encoding genes of different E. coli pathotypes was searched in a large collection of EAEC strains, and a possible association between SPATEs and E. coli phylogroups was investigated. Among 108 typical and 85 atypical EAEC, pic was the most prevalent gene, detected in 47.1% of the strains, followed by sat (24.3%), espI (21.2%), pet (19.2%), sepA (13.5%), sigA (4.1%), eatA (4.1%), vat (1.0%), espP and tsh, detected in one strain (0.5%) each; while epeA and espC were not detected. Phylogenetic analysis demonstrated that 39.9% of the strains belonged to group A, 23.3% to B1, 10.9% to B2, 7.8% to D, 8.8% to E and 1.5% to F. The majority of the SPATE genes were distributed in typical and atypical strains without association with any phylogroup. In addition, pic and pet were strongly associated with typical EAEC and sepA was detected in close association with atypical EAEC. Our data indicate that SPATEs may represent important virulence traits in both subgroups of EAEC.

National Institutes of Health Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Provost's Office for Research of the University of São Paulo Programa de Incentivo a Pesquisa NAPNA-USP, Brazil International Rett Syndrome Foundation Emerald Foundation California Institute for Regenerative Medicine Grant Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Bradykinin is not only important for inflammation and blood pressure regulation, but also involved in neuromodulation and neuroprotection. Here we describe novel functions for bradykinin and the kinin-B2 receptor (B2BkR) in differentiation of neural stem cells. in the presence of the B2BkR antagonist HOE-140 during rat neurosphere differentiation, neuron-specific beta 3-tubulin and enolase expression was reduced together with an increase in glial protein expression, indicating that bradykinin- induced receptor activity contributes to neurogenesis. in agreement, HOE-140 affected in the same way expression levels of neural markers during neural differentiation of murine P19 and human iPS cells. Kinin-B1 receptor agonists and antagonists did not affect expression levels of neural markers, suggesting that bradykinin-mediated effects are exclusively mediated via B2BkR. Neurogenesis was augmented by bradykinin in the middle and late stages of the differentiation process. Chronic treatment with HOE-140 diminished eNOS and nNOS as well as M1-M4 muscarinic receptor expression and also affected purinergic receptor expression and activity. Neurogenesis, gliogenesis, and neural migration were altered during differentiation of neurospheres isolated from B2BkR knock-out mice. Whole mount in situ hybridization revealed the presence of B2BkR mRNA throughout the nervous system in mouse embryos, and less beta 3-tubulin and more glial proteins were expressed in developing and adult B2BkR knock-out mice brains. As a underlying transcriptional mechanism for neural fate determination, HOE-140 induced up-regulation of Notch1 and Stat3 gene expression. Because pharmacological treatments did not affect cell viability and proliferation, we conclude that bradykinin-induced signaling provides a switch for neural fate determination and specification of neurotransmitter receptor expression. Univ São Paulo, Inst Quim, Dept Bioquim, BR-05508000 São Paulo, Brazil Univ Calif San Diego, Dept Pediat, San Diego, CA 92093 USA Univ Calif San Diego, Dept Cellular & Mol Med, San Diego, CA 92093 USA Universidade Federal de São Paulo, Dept Biofis, BR-04023062 São Paulo, Brazil Univ Fed Rio de Janeiro, Inst Ciencias Biomed, BR-21941902 Rio de Janeiro, Brazil Universidade Federal de São Paulo, Dept Biofis, BR-04023062 São Paulo, Brazil National Institutes of Health: 1-DP2-OD006495-01 FAPESP: 2006/61285-9 Provost's Office for Research of the University of São Paulo Programa de Incentivo a Pesquisa: 2011.1.9333.1.3 International Rett Syndrome Foundation: 2517 California Institute for Regenerative Medicine Grant: TR2-01814 Web of Science

Ten-Eleven Translocation (Tet) family of dioxygenases offers a new mechanism for dynamic regulation of DNA methylation and has been implicated in cell lineage differentiation and oncogenesis. Yet their functional roles and mechanisms of action in gene regulation and embryonic development are largely unknown. Here, we report that Xenopus Tet3 plays an essential role in early eye and neural development by directly regulating a set of key developmental genes. Tet3 is an active 5mC hydroxylase regulating the 5mC/5hmC status at target gene promoters. Biochemical and structural studies further reveal a novel DNA binding mode of the Tet3 CXXC domain that is critical for specific Tet3 targeting. Finally, we show that the enzymatic activity and CXXC domain are crucial for Tet3’s biological function. Together, these findings define Tet3 as a novel transcription factor and reveal a molecular mechanism by which the 5mC hydroxylase and DNA binding activities of Tet3 cooperate to control target gene expression and embryonic development.

Objetivou-se, com este estudo, avaliar as correlações entre a digestibilidade aparente e o comportamento ingestivo de novilhas mestiças a pasto. O experimento foi conduzido na fazenda Princesa do Mateiro, município de Ribeirão do Largo, Bahia. Foram utilizadas 20 novilhas com grau de sangue 5/8 Guzerá linhagem leiteira e 3/8 Holandesa, com média de 18 meses de idade e peso corporal médio de 187±13,1 kg. O experimento teve duração de 224 dias e os animais criados em pastejo rotacionado de Brachiaria brizantha "Marandú". Verificou-se uma correlação positiva entre o tempo de pastejo e o coeficiente de digestibilidade da fibra em detergente neutro (CDFDN). Correlações negativas foram verificadas entre o tempo de outras atividades e CDFDN. O tempo de alimentação no cocho apresentou correlações negativas com coeficiente de digestibilidade da matéria seca (CDMS), matéria orgânica (CDMO), carboidratos não fibrosos (CDCNF), carboidratos totais (CDCHOT). Ob-servou-se uma correlação positiva entre o tempo de alimentação total e mastigação total com o CDFDN. O número de períodos de pastejo e ruminação apresentaram correlações negativas com CDMS, CDMO, CDCNF, CDCHOT. Também, verificou-se correlação acima de 0,90 para os nutrientes digestíveis totais (NDT), podendo também ser incluída em equações de predição de consumo. O número de período de alimentação no cocho apresentou correlação negativa com CDMO e coeficiente de digestibilidade de proteína bruta (CDPB). O tempo por período de ruminação apresentou correlação positiva com CDMO, CDCNF, CDCHOT, NDT e CDPB. O tempo por período de outras atividades apresentou correlação positiva com CDCNF e NDT. Correlações negativas foram verificadas entre a taxa de bocados e CDMS, CDMO, CDCNF, CDCHOT e CDPB. O tempo por deglutição correlacionou positivamente com CDMS, CDMO, CDCNF, CDCHOT, NDT, CDPB. A massa do bocado e bocados por deglutição não apre-sentaram correlações com nenhuma das variáveis da digestibilidade. O número de mastigação merícica por dia apresentou correlações negativas com NDT. O tempo de pastejo e ruminação não apresentaram relação com os demais coeficientes de digestibilidade aparente de novilhas mestiças a pasto. O coeficiente da digestibilidade da fibra em detergente neutro provoca variações no tempo de pastejo.

The study was conducted to evaluate the effects of compensatory nutrition on supplemented pasture heifers on ingestive behavior. The experiment was conducted on the farm Princesa do Mateiro, Ribeirao do Largo, Bahia. Twenty 5/8 Guzera Dairy 3/8 Hostein heifers, with an average of 18 months of age and body weight of 187 ± 13.07 kg were used. The experiment lasted 224 days and the animals were maintained in pasture production, rotational grazing Brachiaria brizantha "Marandu". The animals were randomly assigned to each of the treatments: control (T100): Animals fed concentrate to supply 100 % of the requirement of nutrients to gain 750 g/day; compensatory nutrition (CN): animals that alternated the feed level periodically being restricted fed concentrate to meet the requirement of 80 % total digestible nutrients of the control treatment (T80) and refeeding fed concentrate to supply 120 % of the total digestible nutrient requirements of the control treatment (T120). In periods of restriction, the average grazing time was greater for NC (533 vs. 426 minutes) (p 0.05). During refeeding, the number of bits by swallowing, swallowing time and number of bites per day were lower (p

SummarySecreted Wnt morphogens are signaling molecules essential for embryogenesis, pathogenesis, and regeneration and require distinct modifications for secretion, gradient formation, and activity. Whether Wnt proteins can be posttranslationally inactivated during development and homeostasis is unknown. Here we identify, through functional cDNA screening, a transmembrane protein Tiki1 that is expressed specifically in the dorsal Spemann-Mangold Organizer and is required for anterior development during Xenopus embryogenesis. Tiki1 antagonizes Wnt function in embryos and human cells via a TIKI homology domain that is conserved from bacteria to mammals and acts likely as a protease to cleave eight amino-terminal residues of a Wnt protein, resulting in oxidized Wnt oligomers that exhibit normal secretion but minimized receptor-binding capability. Our findings identify a Wnt-specific protease that controls head formation, reveal a mechanism for morphogen inactivation through proteolysis-induced oxidation-oligomerization, and suggest a role of the Wnt amino terminus in evasion of oxidizing inactivation. TIKI proteins may represent potential therapeutic targets.

Connective tissue growth factor (CTGF/CCN2) is a protein of the CCN family that modulates cell–ECM interactions in a variety of cell types. In this study, we investigated the chemotactic and adhesive properties of CCN2 protein in embryonic teratocarcinoma P19 cells. Initially, P19 cells were attracted to CCN2-coated agarose beads. In Boyden chamber experiments, CCN2-containing medium induced a threefold greater migration of P19 cells. CCN2 adhesion properties were studied by using optical tweezers. The specific adhesion times of P19 cells to polystyrene beads coated with laminin, fibronectin, CCN2 and bovine serum albumin were 1.8 ± 0.5s, 2.7 ± 0.4s, 10 ± 2s and 13 ± 2s, respectively, revealing an unexpectedly low adhesive capacity of CCN2 protein for P19 cells. In conclusion, our findings support the chemoattractive role of CCN2 for P19 cells, but not its adhesive role when compared to laminin or fibronectin.

A produção animal em sistemas intensivos tem se ampliado no mundo, sempre visando à elevação de índices produtivos e retorno econômico, respectivamente. Essa verticalização da pecuária tem provocado algumas adaptações de manejo e também nas instalações, que tem alterado o comportamento dos animais, oriundo das restrições tanto visual como espacial do indivíduo. Na pecuária leiteira, a criação de bezerros em baias individuais é um exemplo dessa verticalização, tornando então uma prática comum em muitos países. Sabendo que os bovinos são animais sociais que mostram alta disponibilidade para formar vínculo social com outros indivíduos da mesma espécie, alguns órgãos têm implantado legislações para garantir um mínimo de conforto para esses animais. Um estudo sistemático das preferências sociais dos animais e os efeitos calmantes do animal companheiro de familiaridade diferente são necessários para documentar a resposta à separação. Objetivou-se com esta revisão, apresentar e discutir o efeito do isolamento espacial e/ou visual sobre as respostas produtivas e comportamentais de animais ruminantes.

Objective High-fat diets have been shown to be a risk factor for ulcerative colitis (UC). Omega-6 polyunsaturated fatty acids are considered to increase lipid peroxidation, while the omega-3 polyunsaturated fatty acid exerts a chemopreventative effect. We evaluated the effect of high-fat diets (20%) enriched with fish or soybean oil on colonic inflammation and DNA damage in dextran sulfate sodium-induced colitis. Methods Male Wistar rats (28–30 days) were fed an American Institute of Nutrition (AIN)-93 diet for 47 days and divided into five groups: control normal fat non-colitic (C) or control colitis (CC), high soybean fat group (HS) colitis, high fish fat group colitis, or high-fat soybean plus fish oil colitis. UC was induced from day 35 until day 41 by 3% dextran sulfate sodium. On day 47, the rats were anesthetized; blood samples collected for corticosterone determination, and the distal colon was excised to quantify interleukin-4 (IL-4), IL-10, and interferon-gamma levels, myeloperoxidase activity, histological analyses, and DNA damage. The disease activity index was recorded daily. Results The disease activity index, histological analysis, myeloperoxidase activity, IL-4, interferon-gamma, and corticosterone levels did not differ among the colitic groups. IL-10 was significantly increased by the high fish fat group diet in relation to HS, but only the high soybean-fish fat diet increased the IL-10/IL-4 ratio (anti-inflammatory/pro-inflammatory) to levels closer to the C group and reduced DNA damage compared to the HS group ( P Conclusion The data show that high-fat diets did not exacerbate UC and suggest that the soybean and fish oil mixture, more than the fish oil alone, could be a complementary therapy to achieve a cytokine balance in UC.

Background: Asynchronous electrical activation induced by right ventricular (RV) pacing can cause several abnormalities in left ventricular (LV) function. However, the effect of ventricular pacing on RV function has not been well established. We evaluated RV function in patients undergoing long-term RV pacing. Methods: Eighty-five patients and 24 healthy controls were included. After pacemaker implantation, conventional echocardiography and strain imaging were used to analyze RV function. Strain imaging measurements included peak systolic strain and strain rate. LV function and ventricular dyssynchrony by tissue Doppler imaging (TDI) were assessed. Intra- and interobserver variabilities of TDI parameters were tested on 15 randomly selected cases. Results: All patients were in New York Heart Association functional class I or II and percentage of ventricular pacing was 96 ± 4%. RV apical induced interventricular dyssynchrony in 49 patients (60%). LV dyssynchrony was found in 51 patients (60%), when the parameter examined was the standard deviation of the time to peak myocardial systolic velocity of all 12 segments greater than 34 ms. Likewise, septal-to-lateral delay ≥65 ms was found in 31 patients (36%). All echocardiographic indexes of RV function were similar between patients and controls (strain: −22.8 ± 5.8% vs −22.1 ± 5.6%, P = 0.630; strain rate: −1.47 ± 0.91 s−1 vs −1.42 ± 0.39 s−1, P = 0.702). Intra- and interobserver variability for RV strain was 3.1% and 5.3%, and strain rate was 1.3% and 2.1%, respectively. Conclusions: In patients with standard pacing indications, RV apical pacing did not seem to affect RV systolic function, despite induction of electromechanical dyssynchrony. (PACE 2011; 34:155–162)

A pneumonia é uma doença infecciosa que atinge os alvéolos pulmonares, ocasionada, principalmente, por bactérias, vírus, fungos e parasitas. Dentre as principais bactérias, Haemophilus influenzae, Klebsiella pneumonia, Streptococcus pneumoniae, Staphylococcus aureus e as do grupo das Gram-negativas não fermentadoras de glicose, a exemplo de Acinetobacter spp. e Pseudomonas aeruginosa, são as mais isoladas. Estima-se que 450 milhões de casos de pneumonia ocorram por ano em todo o mundo e o Brasil é protagonista na lista dos países com maior número de casos. Pacientes que fazem uso de ventilação mecânica têm até 21 vezes mais chances de desenvolver a doença e 20% daqueles que utilizam a técnica por mais de 48 horas desenvolverão a infecção. Desta forma, esta revisão teve por objetivo fazer uma avaliação crítica sistematizada da literatura sobre a doença, aspectos epidemiológicos, fisiopatologia e avanços no diagnóstico. Este estudo observou que houve uma evolução nos métodos de diagnóstico, resultando na diminuição expressiva do número de mortes. No entanto, para que a queda no número de acometidos se mantenha, a correta prescrição e utilização da antibioticoterapia são imprescindíveis.

The Wnt pathway regulates multiple biological and pathological processes including angiogenesis and inflammation. Here we identified a unique inhibitor of the Wnt pathway, SERPINA3K, a serine proteinase inhibitor with anti-inflammatory and angiogenic activities. SERPINA3K blocked the Wnt pathway activation induced by a Wnt ligand and by diabetes. Coprecipitation and ligand binding assay showed that SERPINA3K binds to low-density lipoprotein receptor-like protein 6 (LRP6) with a K d of 10 nM, in the range of its physiological concentration in the retina. Under the same conditions, SERPINA3K did not bind to the frizzled (Fz) receptor or low-density lipoprotein receptor. Further, SERPINA3K bound to LRP6 at the extracellular domain and blocked its dimerization with the Fz receptor induced by a Wnt ligand. The antagonizing activity of SERPINA3K to LRP6 was further confirmed by Xenopus axis duplication assay. These results suggest that SERPINA3K is a high-affinity, endogenous antagonist of LRP6. The blockade of Wnt signaling may represent a unifying mechanism for the anti-inflammatory and anti-angiogenic effects of SERPINA3K.

The cultivated area devoted to rice crops (Oryza sativa) in Brazil has expanded in recent years, along with improvements in yield. This has prompted the continuous exchange of material from active germplasm banks between government breeding programs. When placed in a greenhouse for multiplication purposes, rice accessions resulting from exchanges between USDA-ARS (United States Department of Agriculture - Agricultural Research Service) showed little or no germination. The accessions were sent to the tissue culture laboratory of BAG-CNPAF (Embrapa National Rice and Bean Research Center) to recover embryos by aseptically removing the seed coat of 10 seeds from each accession. Bacterial colony growth concomitant to germination inhibition was observed in some accessions, from which four bacterial isolates were obtained. Made available in DSpace on 2017-12-26T23:23:54Z (GMT). No. of bitstreams: 1 CNPAF2018pdv102n1.pdf: 213919 bytes, checksum: c3d290b54a0663e654d67513d579c97c (MD5) Previous issue date: 2017-12-18 Disease notes.