Quantitative Proteomic Analysis Reveals Changes in the Benchmark Corynebacterium pseudotuberculosis Biovar Equi Exoproteome after Passage in a Murine Host

Corynebacterium pseudotuberculosis biovar equi is the etiologic agent of ulcerative lymphangitis. To investigate proteins that could be related to the virulence of this pathogen, we combined an experimental passage process using a murine model and high-throughput proteomics with a mass spectrometry, data-independent acquisition (LC-MSE) approach to identify and quantify the proteins released into the supernatants of strain 258_equi. To our knowledge, this approach allowed characterization of the exoproteome of a C. pseudotuberculosis equi strain for the first time. Interestingly, the recovery of this strain from infected mouse spleens induced a change in its virulence potential, and it became more virulent in a second infection challenge. Proteomic screening performed from culture supernatant of the control and recovered conditions revealed 104 proteins that were differentially expressed between the two conditions. In this context, proteomic analysis of the recovered condition detected the induction of proteins involved in bacterial pathogenesis, mainly related to iron uptake. In addition, KEGG enrichment analysis showed that ABC transporters, bacterial secretion systems and protein export pathways were significantly altered in the recovered condition. These findings show that secretion and secreted proteins are key elements in the virulence and adaptation of C. pseudotuberculosis. Collectively, bacterial pathogenesis-related proteins were identified that contribute to the processes of adherence, intracellular growth and evasion of the immune system. Moreover, this study enhances our understanding of the factors that may influence the pathogenesis of C. pseudotuberculosis.Fil: Marques Da Silva, Wanderson. Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas; Brasil. Institut National de la Recherche Agronomique; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carvalho, Rodrigo D. De Oliveira. Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas; BrasilFil: Dorella, Fernanda A.. Universidade Federal de Minas Gerais; BrasilFil: Folador, Edson L.. Universidade Federal da Paraíba. Centro de Biotecnologia; BrasilFil: Souza, Gustavo H. M. F.. Waters Corporation; BrasilFil: Pimenta, Adriano M. C.. Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas; BrasilFil: Figueiredo, Henrique C. P.. Universidade Federal de Minas Gerais; BrasilFil: Le Loir, Yves. Institut National de la Recherche Agronomique; FranciaFil: Silva, Artur. Universidade Federal do Pará; BrasilFil: Azevedo, Vasco. Universidade Federal de Minas Gerais. Instituto de Ciências Biológicas; Brasi

Lipid oxidation of a meat product packaged with poly (lactic acid)/clay nanocomposites

Introduction: Polylactic acid or polylactide (PLA, Poly) is a biodegradable thermoplastic aliphatic polyester derived from renewable resources, such as corn starch, tapioca roots, chips or starch, or sugarcane. Biopolymer nanocomposites are of great interest to the packaging industry as they can overcome the limitations of biopolymers compared to synthetic polymers. In the last two decades, the nanocomposites have been studied intensively, once the addition of fillers such as organoclays, in particular, montmorillonite (MMT), can improve rheological, thermal and mechanical properties of the biopolymers (Jollands M. et al. 2010). The presence of MMT can lead to materials which generally exhibit great property enhancements, mainly due to its intercalation or exfoliation into the polymer chains. In this work, PLA was incorporated with 5% (w/w) Cloisite Na+ prepared through a two-step process: first extrusion of pellets and secondly melted matter was pressed. The nanocomposite was used to pack a model food (salami) in order to evaluate of the ability of the new packaging to inhibit lipid oxidation. Thiobarbituric Acid Reactive Substances (TBARS) assay was used to evaluate the lipid oxidation stage. This assay allows to measure malondialdehyde (MDA) content, which is formed in the lipid oxidation of polyunsaturated fatty acids. Material and Methods: Packaged salami was homogenized with trichloroacetic acid (10 %) in 0.02 M of orthophosphoric acid and the solution was filtered. The filtered solution was homogenized with thiobarbituric acid aqueous solution (0.02 M) and heated at 100 °C for 40 min. Solutions were cooled down and absorbance was measured at 530 nm. Results were expressed as mg MDA per kg of salami. Results and Discussion: Salami slices were packaged with a control film and with the nanocomposite and analysed at initial time and after 15, 30, 60 and 90 days of contact. Results showed that salami packaged with the nanocomposite presented lower amount of MDA after all contact periods, except after 60 days, where there were no differences between control and nanofilm. Conclusion: Although the results showed that the new nanocomposite tends to reduce the production of MDA, further studies should be carried out to confirm the inhibition of lipid oxidation, such as the peroxide index, p-anisidine value, or the monitorization of a lipid oxidation indicator like hexanal.Project “Development of methodologies for the evaluation of polymeric food packaging components and determination of their structural and mechanical properties” (2016DAN 1289)N/

Effect of Various Proteins on Different Molecular Weight Proanthocyanidin Fractions of Red Wine during Wine Fining

The effect of several proteins on three main wine proanthocyanidin containing fractions with the mean degree of polymerization (mDP) of 1.5 (FI), 3.4 (FII), and 4.9 (FIII) was studied. Although casein and potassium caseinate showed similar molecular weight (MW) distribution, casein decreased the FI fraction more than the twice as effectively as potassium caseinate. A gelatin with a medium MW polydispersion induced a similar decrease (~20%) in all tannin fractions. A gelatin with low MW primarily removed the tannin fractions of lower mDP (FI and FII), while a gelatin with a higher MW had a minor effect (5%) on the fraction of higher mDP (FIII). Neither of the two studied isinglasses reduced the FII fraction. The tannins of FI and FIII were removed by swim bladder isinglass twice as effectively as by fish skin isinglass. For the mDP of fined wines, egg albumin induced a decrease on mDP of 24% for the more polymerized tannin fraction (FIII); although within all assays there was a decrease ranging from 6 to 14%info:eu-repo/semantics/publishedVersio

Humibacter albus gen. nov., sp. nov., isolated from sewage sludge compost

A bacterial strain isolated from sewage sludge compost, strain SC-083T, was characterized. The isolate was a motile, Gram-positive, short rod, forming coryneform V-shaped cells during the early stages of growth. The organism was strictly aerobic and able to grow between 22 and 36 °C and between pH 5.5 and 8.0. The predominant fatty acids were cyclohexyl-C17 : 0, anteiso-C17 : 0 and iso-C16 : 0, the major respiratory quinones were menaquinone 11 (MK-11) and 12 (MK-12), and the genomic DNA G+C content was 68 mol%. The peptidoglycan contained the diagnostic diamino acids ornithine and 2,4-diaminobutyric acid and was of acetyl type. The 16S rRNA gene sequence analysis indicated that this isolate belongs to the family Microbacteriaceae with the type strains of the species Leifsonia xyli (96 % gene sequence similarity), Leifsonia shinshuensis (96 %), Leifsonia naganoensis (95 %), Leifsonia aquatica (95 %), Agromyces ramosus (95 %) and Curtobacterium citreum (95 %) among the closest phylogenetic neighbours. The phylogenetic analysis and phenetic characteristics support the proposal of a new genus and a novel species, with the name Humibacter albus gen. nov., sp. nov. The type strain of Humibacter albus is SC-083T (=DSM 18994T =CCUG 54538T =LMG 23996T

Effects of combined training on metabolic profile, lung function, stress and quality of life in sedentary adults: a study protocol for a randomized controlled trial

Background Both physical inactivity and sedentary behavior are considered modifiable risk factors for chronic diseases and all-cause mortality. Adult office-workers spend most of their working day in sedentary behaviors, so they are particularly at high risk of developed chronic diseases (e.g., cardiovascular diseases, metabolic disorders like diabetes mellitus, . . .). It seems important to promote behavioral changes that could prevent or delay metabolic disease development. Evidence supports the use of exercise programs, however, to date there are several knowledge gaps and inconsistencies in the literature regarding the effects of Combined Training (i.e., aerobic plus resistance training) in sedentary healthy adults. This paper outlines an RCT designed to evaluate the effects of a 16-week combined training program on biochemical and immune markers of metabolic disease, lung function, salivary stress hormones and subjective quality of life (primary outcomes), as well as on body composition and physical fitness (secondary outcomes) in sedentary middle-aged office-workers. Furthermore, we aimed to assess the associations between the changes promoted by the exercise program and the different variables studied. Methods and design This is a single-blinded two-arm RCT with parallel groups. A minimum of healthy 40 officeworkers aged 40–64 years will be recruited to engage in a 16-week intervention study. After baseline assessments, participants will be randomized to one of the two groups: (1) combined training group or (2) control group. Baseline assessments will be repeated after 8 weeks of intervention (mid-testing) and upon completion of the intervention (post-testing). Discussion This RCT involves a multi-disciplinary approach and seems to be a relevant contribution to understanding the potential role of combined training in improving the metabolic profile, lung function, stress, and quality of life in adults. The results can provide important insights for clinical recommendations and for the optimization of strategies to prevent metabolic disorders in adults with sedentary jobs.info:eu-repo/semantics/publishedVersio

Protective role of new nitrogen compounds on ROS/RNS-mediated damage to PC12 cells

Reactive oxygen (ROS) and nitrogen (RNS) species are known to be involved in many degenerative diseases. This study reports four new nitrogen compounds from organic synthesis, identified as FMA4, FMA7, FMA762 and FMA796, which differ mainly by the number of hydroxyl groups within their phenolic unit. Their potential role as antioxidants was evaluated in PC12 cells by assessing their protection against oxidative and nitrosative insults. The four compounds, and particularly FMA762 and FMA796, were able to protect cells against lipid peroxidation and intracellular ROS/RNS formation to a great extent. Their protective effects were likely mediated by their free radicals scavenging ability, as they appeared to be involved neither in the induction of natural antioxidant enzymes like GSH-PX and SOD, nor in the inhibition of NOS. Nevertheless, these results suggest a promising potential for these compounds as ROS/RNS scavengers in pathologies where oxidative/ nitrosative stress are involved.Fundação para a Ciência e Tecnologia (FCT

Global vs. local nonlinear optimization techniques for human-like movement of an anthropomorphic robot

In this paper a comparison between using global and local optimization techniques for solving the problem of generating human-like arm and hand movements for an anthropomorphic dual arm robot is made. Although the objective function involved in each optimization problem is convex, there is no evidence that the admissible regions of these problems are convex sets. For the sequence of movements for which the numerical tests were done there were no significant differences between the optimal solutions obtained using the global and the local techniques. This suggests that the optimal solution obtained using the local solver is indeed a global solution

Monitorization of hexanal as lipid oxidation indicator in a processed meat product packaged with poly(lactic acid)/clay nanocomposite films

One of the most detrimental processes in fatty foodstuffs is lipid oxidation, which occurs during production and storage, and influences food composition and safety. Through the analysis of volatile lipid oxidation products we can have an insight into the oxidation, and some volatiles, such as hexanal, which can be markers of undergoing oxidation processes. Hexanal is formed when fatty acids are oxidized and is one of many well-documented aromatic components that contributes to flavour and aroma in common food products containing fatty acids. During the last decade, the interest in polymer layered silicate (PLS) nanocomposites has rapidly increased due to their potential for enhancing physical, chemical, and mechanical properties of conventional materials. Polymer nanocomposites are represented by a polymeric matrix reinforced with nanoscale fillers, among them the most common silicate clays are represented by montmorillonite (MMT), which is naturally occurring and readily available in large quantities. The presence of MMT can lead to materials which generally exhibit great property enhancements, mainly due to its intercalation or exfoliation into the polymer chains. In this work natural MMT Cloisite Na+ was incorporated in PLA. The PLA/Cloisite® Na+ films were prepared through a two-step process. In the first step, PLA pellets were fed into a corotating laboratory twin-screw extruder at 170 °C and 50 rpm for 2 min. Subsequently, Cloisite® Na+ powder (5%, w/w) were added and mixed. After extrusion, the melted matter was then pressed with a P300P hot press at 170 °C and 100 bar to obtain the PLA/Cloisite® Na+ films. Salami slices were packaged with PLA-OMMT film and with a control film (PLA). After different storage times (0, 15, 30, 60 and 90 days), salami slices were analysed regarding their hexanal content. The hexanal derivatization was performed in a solution of 2,4-dinitrophenylhydrazine in sulfuric acid during 4 h in the dark, and the hexanal extraction was performed with n-hexane and evaporation till dryness. The residue was dissolved in methanol, filtered and analysed. The quantification of hexanal was performed by Ultra High Performance Liquid Chromatography coupled with Diode Array Detector at 365 nm, with a Pre-column AcquityTM UPLC® BEH C18 (2.1 x 5 mm, 1.7 μm particle size) and a column AcquityTM UPLC® BEH C18 (2.1 × 50 mm, 1.7 μm particle size), the mobile-phase was acetonitrile-water (75:25, v/v). The amount of hexanal in packaged salami decreased in the first 60 days of storage. In this period of time the hexanal content of the salami packaged with the PLA/Cloisite® Na+ films was lower than the salami packaged with control film, except after 15 days of storage, where there was no difference between two films. After 90 days of storage, the amount of hexanal in the samples increased, although it was higher in the samples packaged with control film (94.7 ± 6.02 μg/100g salami) than salami packaged with PLA/Cloisite® Na+ films (65.1 ± 6.12 μg/100g salami). The presence of MMT in the polymer film can reduce the lipid oxidation of processed meat products, extending their shelf life. Further studies to evaluate differences between PLA and the nanocomposite (PLA-5%Cloisite®Na+) in what regards to the mechanical and barrier properties are in progress.This work was supported by the research project “Labelling and tracking of nanoclay from food packaging nanocomposites: a food safety issue – NanoPack4Food” (2014DAN1019) under the Cooperative Programme of the Agreement on Scientific Cooperation between National Research Council of Italy (CNR) and Foundation for Science and Technology of Portugal (FCT)N/