Machine intelligence decrypts β-lapachone as an allosteric 5-lipoxygenase inhibitor.

Using machine learning, targets were identified for β-lapachone. Resorting to biochemical assays, β-lapachone was validated as a potent, ligand efficient, allosteric and reversible modulator of 5-lipoxygenase (5-LO). Moreover, we provide a rationale for 5-LO modulation and show that inhibition of 5-LO is relevant for the anticancer activity of β-lapachone. This work demonstrates the power of machine intelligence to deconvolute complex phenotypes, as an alternative and/or complement to chemoproteomics and as a viable general approach for systems pharmacology studies

Validação conceitual das características definidoras de diagnósticos de enfermagem respiratórios em neonatos

OBJECTIVE:To develop and validate conceptual and operational definitions for the defining characteristics of the respiratory nursing diagnoses, ineffective breathing pattern, impaired gas exchange and impaired spontaneous ventilation, in newborns.METHODS:This was a methodological study of conceptual validation of the defining characteristics of three respiratory nursing diagnoses, by consensus analysis of a committee of five specialist nurses, and then a group of five non-nursing professionals, using the Delphi technique.RESULTS:After two rounds of evaluation, consensus was obtained that was equal to or greater than 80% on all of the definitions, which were then considered validated.CONCLUSION:The definitions developed for the defining characteristics of three nursing diagnoses were validated with a high level of consensus.OBJETIVO:Elaborar e validar definições conceituais e operacionais para as características definidoras dos diagnósticos de enfermagem respiratórios, Padrão Respiratório Ineficaz, Troca de Gases Prejudicada e Ventilação Espontânea Prejudicada em recém-nascidos.MÉTODOS:Estudo metodológico, de validação conceitual das características definidoras dos três diagnósticos de enfermagem respiratórios por meio da análise de consenso de um comitê de cinco enfermeiras especialistas e de cinco profissionais não enfermeiros, utilizando a técnica Delphi.RESULTADOS:Após duas rodadas de avaliação, obteve-se consenso igual ou superior a 80% na totalidade das definições, sendo consideradas validadas.CONCLUSÃO:As definições elaboradas para as características definidoras dos três diagnósticos de enfermagem foram validadas com elevado grau de consenso.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de EnfermagemUNIFESP, EPESciEL

Study of the impact of the post-MS evolution of the host star on the orbits of close-in planets. I. Sample definition and physical properties

Context: To date, more than 30 planets have been discovered around giant stars, but only one of them has been found to be orbiting within 0.6 AU from the host star, in direct contrast to what is observed for FGK dwarfs. This result suggests that evolved stars destroy/engulf close-in planets during the red giant phase. Aims: We are conducting a radial velocity survey of 164 bright G and K giant stars in the southern hemisphere with the aim of studying the effect of the host star evolution on the inner structure of planetary systems. In this paper we present the spectroscopic atmospheric parameters (\Teff, \logg, $\xi$, [Fe/H]) and the physical properties (mass, radius, evolutionary status) of the program stars. In addition, rotational velocities for all of our targets were derived. Methods: We used high resolution and high S/N spectra to measure the equivalent widths of many Fe{\sc\,i} and Fe{\sc\,ii} lines, which were used to derive the atmospheric parameters by imposing local thermodynamic and ionization equilibrium. The effective temperatures and metallicities were used, along with stellar evolutionary tracks to determine the physical properties and evolutionary status of each star. Results: We found that our targets are on average metal rich and they have masses between $\sim$\,1.0\,M$_\odot$ and 3.5\,M$_\odot$. In addition, we found that 122 of our targets are ascending the RGB, while 42 of them are on the HB phase.Comment: Accepted for publication in A&

Cellular responses of Candida albicans to phagocytosis and the extracellular activities of neutrophils are critical to counteract carbohydrate starvation, oxidative and nitrosative stress

Acknowledgments We thank Alexander Johnson (yhb1D/D), Karl Kuchler (sodD/D mutants), Janet Quinn (hog1D/D, hog1/cap1D/D, trx1D/D) and Peter Staib (ssu1D/D) for providing mutant strains. We acknowledge helpful discussions with our colleagues from the Microbial Pathogenicity Mechanisms Department, Fungal Septomics and the Microbial Biochemistry and Physiology Research Group at the Hans Kno¨ll Institute (HKI), specially Ilse D. Jacobsen, Duncan Wilson, Sascha Brunke, Lydia Kasper, Franziska Gerwien, Sea´na Duggan, Katrin Haupt, Kerstin Hu¨nniger, and Matthias Brock, as well as from our partners in the FINSysB Network. Author Contributions Conceived and designed the experiments: PM HW IMB AJPB OK BH. Performed the experiments: PM CD HW. Analyzed the data: PM HW IMB AJPB OK BH. Wrote the paper: PM HW OK AJPB BH.Peer reviewedPublisher PD

Leishmania amazonensis Arginase Compartmentalization in the Glycosome Is Important for Parasite Infectivity

In Leishmania, de novo polyamine synthesis is initiated by the cleavage of L-arginine to urea and L-ornithine by the action of arginase (ARG, E.C. 3.5.3.1). Previous studies in L. major and L. mexicana showed that ARG is essential for in vitro growth in the absence of polyamines and needed for full infectivity in animal infections. The ARG protein is normally found within the parasite glycosome, and here we examined whether this localization is required for survival and infectivity. First, the localization of L. amazonensis ARG in the glycosome was confirmed in both the promastigote and amastigote stages. As in other species, arg− L. amazonensis required putrescine for growth and presented an attenuated infectivity. Restoration of a wild type ARG to the arg− mutant restored ARG expression, growth and infectivity. In contrast, restoration of a cytosol-targeted ARG lacking the glycosomal SKL targeting sequence (argΔSKL) restored growth but failed to restore infectivity. Further study showed that the ARGΔSKL protein was found in the cytosol as expected, but at very low levels. Our results indicate that the proper compartmentalization of L. amazonensis arginase in the glycosome is important for enzyme activity and optimal infectivity. Our conjecture is that parasite arginase participates in a complex equilibrium that defines the fate of L-arginine and that its proper subcellular location may be essential for this physiological orchestration

Formation of Lipofuscin-Like Autofluorescent Granules in the Retinal Pigment Epithelium Requires Lysosome Dysfunction

PURPOSE: We aim to characterize the pathways required for autofluorescent granule (AFG) formation by RPE cells using cultured monolayers. METHODS: We fed RPE monolayers in culture with a single pulse of photoreceptor outer segments (POS). After 24 hours the cells started accumulating AFGs that were comparable to lipofuscin in vivo. Using this model, we used a variety of light and electron microscopical techniques, flow cytometry and Western blot to analyze the formation of AFGs. We also generated a mutant RPE line lacking cathepsin D by gene editing. RESULTS: AFGs seem to derive from incompletely digested POS-containing phagosomes and after 3 days are surrounded by a single membrane positive for lysosome markers. We show by various methods that lysosome-phagosome fusion is required for AFG formation, and that impairment of lysosomal pH or catalytic activity, particularly cathepsin D activity, enhances AF accumulation. CONCLUSIONS: We conclude that lysosomal dysfunction results in incomplete POS degradation and enhanced AFG accumulation

A Randomized Double-Blind Study Comparing the Efficacy and Safety of Orlistat Versus Placebo in Obese Patients with Mild to Moderate Hypercholesterolemia

INTRODUCTION: Obesity is a chronic disease and a serious health problem that leads to increased prevalence of diabetes, hypertension, dyslipidemia and gallbladder disease. OBJECTIVE: To evaluate the efficacy of orlistat for weight loss and improved lipid profile compared to placebo in obese patients with hypercholesterolemia, treated over a period of 6 months. METHODOLOGY: In a 6-month, multicenter (10 centers in Portugal), double-blind, parallel, placebo-controlled study, 166 patients, aged 18-65 years, body mass index (BMI) > or = 27 kg/m2, LDL cholesterol > 155 mg/dl, were randomized to a reduced calorie diet (600 kcal/day deficit) plus orlistat three times a day or placebo. Exclusion criteria included triglycerides > 400 mg/dl, severe cardiovascular disease, uncontrolled hypertension, type 1 or 2 diabetes under pharmacological treatment, and gastrointestinal or pancreatic disease. RESULTS: The mean difference in weight from baseline was 5.9% (5.6 kg) in the orlistat group vs. 2.3% (2.2 kg) in the placebo group. In the orlistat group 49% of patients achieved 5-10% weight loss and 8.8% achieved > 10%. The orlistat group showed a significant reduction in total and LDL cholesterol, with similar changes for HDL in both treatment groups. The frequency of gastrointestinal adverse events was slightly higher in the orlistat group than in the placebo group, leading to discontinuation in 7 patients. CONCLUSION: Treatment with orlistat plus a reduced calorie diet for 6 months achieved significant reductions in weight, BMI and lipid parameters

A Phosphoproteomic Approach towards the Understanding of the Role of TGF-β in Trypanosoma cruzi Biology

Transforming growth factor beta (TGF-β) plays a pivotal role in Chagas disease, not only in the development of chagasic cardiomyopathy, but also in many stages of the T. cruzi life cycle and survival in the host cell environment. The intracellular signaling pathways utilized by T. cruzi to regulate these mechanisms remain unknown. To identify parasite proteins involved in the TGF-β response, we utilized a combined approach of two-dimensional gel electrophoresis (2DE) analysis and mass spectrometry (MS) protein identification. Signaling via TGF-β is dependent on events of phosphorylation, which is one of the most relevant and ubiquitous post-translational modifications for the regulation of gene expression, and especially in trypanosomatids, since they lack several transcriptional control mechanisms. Here we show a kinetic view of T. cruzi epimastigotes (Y strain) incubated with TGF-β for 1, 5, 30 and 60 minutes, which promoted a remodeling of the parasite phosphorylation network and protein expression pattern. The altered molecules are involved in a variety of cellular processes, such as proteolysis, metabolism, heat shock response, cytoskeleton arrangement, oxidative stress regulation, translation and signal transduction. A total of 75 protein spots were up- or down-regulated more than twofold after TGF-β treatment, and from these, 42 were identified by mass spectrometry, including cruzipain–the major T. cruzi papain-like cysteine proteinase that plays an important role in invasion and participates in the escape mechanisms used by the parasite to evade the host immune system. In our study, we observed that TGF-β addition favored epimastigote proliferation, corroborating 2DE data in which proteins previously described to be involved in this process were positively stimulated by TGF-β