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

Metarhizium anisopliae blastospores are highly virulent to adult Aedes aegypti, an important arbovirus vector

BackgroundThe use of entomopathogenic fungi (EPF) for the control of adult mosquitoes is a promising alternative to synthetic insecticides. Previous studies have only evaluated conidiospores against adult mosquitoes. However, blastospores, which are highly virulent against mosquito larvae and pupae, could also be effective against adults.MethodsMetarhizium anisopliae (ESALQ 818 and LEF 2000) blastospores and conidia were first tested against adult Aedes aegypti by spraying insects with spore suspensions. Blastospores were then tested using an indirect contact bioassay, exposing mosquitoes to fungus-impregnated cloths. Virulence when using blastospores suspended in 20% sunflower oil was also investigated.ResultsFemale mosquitoes sprayed with blastospores or conidia at a concentration of 108 propagules ml−1 were highly susceptible to both types of spores, resulting in 100% mortality within 7 days. However, significant differences in virulence of the isolates and propagules became apparent at 107 spores ml−1, with ESALQ 818 blastospores being more virulent than LEF 2000 blastospores. ESALQ 818 blastospores were highly virulent when mosquitoes were exposed to black cotton cloths impregnated with blastospores shortly after preparing the suspensions, but virulence declined rapidly 12 h post-application. The addition of vegetable oil to blastospores helped maintain virulence for up to 48 h.ConclusionThe results showed that blastospores were more virulent to adult female Ae. aegypti than conidia when sprayed onto the insects or applied to black cloths. Vegetable oil helped maintain blastospore virulence. The results show that blastospores have potential for use in integrated vector management, although new formulations and drying techniques need to be investigated

An IT service management literature review: Challenges, benefits, opportunities and implementation practices

Information technology (IT) service management is considered a collection of frameworks that support organizations managing services. The implementation of these kinds of frameworks is constantly increasing in the IT service provider domain. The main objective is to define and manage IT services through its life cycle. However, from observing the literature, scarcely any research exists describing the main concepts of ITSM. Many organizations still struggle in several contexts in this domain, mainly during implementation. This research aims to develop a reference study detailing the main concepts related with ITSM. Thus, a systematic literature review is performed. In total, 47 articles were selected from top journals and conferences. The benefits, challenges, opportunities, and practices for ITSM implementation were extracted, critically analysed, and then discussed.info:eu-repo/semantics/publishedVersio

Dynamics of Snoring Sounds and Its Connection with Obstructive Sleep Apnea

Snoring is extremely common in the general population and when irregular may indicate the presence of obstructive sleep apnea. We analyze the overnight sequence of wave packets --- the snore sound --- recorded during full polysomnography in patients referred to the sleep laboratory due to suspected obstructive sleep apnea. We hypothesize that irregular snore, with duration in the range between 10 and 100 seconds, correlates with respiratory obstructive events. We find that the number of irregular snores --- easily accessible, and quantified by what we call the snore time interval index (STII) --- is in good agreement with the well-known apnea-hypopnea index, which expresses the severity of obstructive sleep apnea and is extracted only from polysomnography. In addition, the Hurst analysis of the snore sound itself, which calculates the fluctuations in the signal as a function of time interval, is used to build a classifier that is able to distinguish between patients with no or mild apnea and patients with moderate or severe apnea

Listening to Music in the First, but not the Last 1.5 km of a 5-km Running Trial Alters Pacing Strategy and Improves Performance

We examined the effects of listening to music on attentional focus, rating of perceived exertion (RPE), pacing strategy and performance during a simulated 5-km running race. 15 participants performed 2 controlled trials to establish their best baseline time, followed by 2 counterbalanced experimental trials during which they listened to music during the first (M-start) or the last (M-finish) 1.5 km. The mean running velocity during the first 1.5 km was significantly higher in M-start than in the fastest control condition (p < 0.05), but there was no difference in velocity between conditions during the last 1.5 km (p > 0.05). The faster first 1.5 m in M-start was accompanied by a reduction in associative thoughts compared with the fastest control condition. There were no significant differences in RPE between conditions (p > 0.05). These results suggest that listening to music at the beginning of a trial may draw the attentional focus away from internal sensations of fatigue to thoughts about the external environment. However, along with the reduction in associative thoughts and the increase in running velocity while listening to music, the RPE increased linearly and similarly under all conditions, suggesting that the change in velocity throughout the race may be to maintain the same rate of RPE increase.Australian Department of Education, Employment and Workplace RelationsAustralian Department of Education, Employment and Workplace Relation

PLANNING PROCESS OF PILOT BATCH PRODUCTION OF AN INNOVATIVE DRUG FOR CLINICAL TRIAL IN A PHARMACEUTICAL INDUSTRY

Purpose – Clinical trials are the most critical step in the process of drug development and evaluation to bring a new drug to market. The purpose of this article was to show an approach of the planning and production of a new innovative drug for a clinical study, based on the presentation of decision-making process in a national pharmaceutical industry. Design / methodology / approach - Through a case-study methodology, it was described the pilot batch production planning of a new drug for clinical trial, focusing on how the company evaluates the adequacy of the available systems at the manufacturing plant and how they use them in drug production planning process. Findings – A previous planning for clinical trial supplies production is determinant to decide the order, amount and timing of the products to be produced when the manufacturing plant is shared with the production of commercial products. Also, even in a small pilot batch production, there is a substantial waste of supplies during the process. Research Limitations / Implications – This study showed the production planning process of one investigational product with the recruitment of few patients for the clinical trial. However, the number of patients enrolled can reach thousands in many clinical trials, and it does need a more complex production planning to avoid wastes and try to reduce the process costs. Practical Implications - This article provides a picture of the production planning of clinical trial supplies chain under uncertainty and the decisions that affect the large-scale production of commercial drugs and the pilot batch production of experimental drugs. Originality / value - Although the results of clinical trials is the most significant source of uncertainty in the development process of any new drug, a good clinical supply planning and processes management can avoid or attenuate the imminent risk of process failure.

Trajetória das Reformas Institucionais da Indústria Elétrica Brasileira e Novas Perspectivas de Mercado

This paper aims at presenting electricity negotiation alternatives in Brazil, which have resulted from a decade of structural and institutional reforms. With the purpose of achieving that, the specific characteristics of the Brazilian electric industry, the international experience and the historical restructuring processes witnessed by Brazil are going to be described, so that mistakes and solutions can be high lightened. As a consequence, it is expected that a wider picture not only of the new commercial and institutional environment in the country but also of the new perspectives faced by the agents in the electric sector is created.Regulaçãao Econômica, Indústria de Energia Elétrica Brasileira

A combined approach for comparative exoproteome analysis of Corynebacterium pseudotuberculosis

Background: Bacterial exported proteins represent key components of the host-pathogen interplay. Hence, we sought to implement a combined approach for characterizing the entire exoproteome of the pathogenic bacterium Corynebacterium pseudotuberculosis, the etiological agent of caseous lymphadenitis (CLA) in sheep and goats. Results: An optimized protocol of three-phase partitioning (TPP) was used to obtain the C. pseudotuberculosis exoproteins, and a newly introduced method of data-independent MS acquisition (LC-MSE) was employed for protein identification and label-free quantification. Additionally, the recently developed tool SurfG+ was used for in silico prediction of sub-cellular localization of the identified proteins. In total, 93 different extracellular proteins of C. pseudotuberculosis were identified with high confidence by this strategy; 44 proteins were commonly identified in two different strains, isolated from distinct hosts, then composing a core C. pseudotuberculosis exoproteome. Analysis with the SurfG+ tool showed that more than 75% (70/93) of the identified proteins could be predicted as containing signals for active exportation. Moreover, evidence could be found for probable non-classical export of most of the remaining proteins. Conclusions: Comparative analyses of the exoproteomes of two C. pseudotuberculosis strains, in addition to comparison with other experimentally determined corynebacterial exoproteomes, were helpful to gain novel insights into the contribution of the exported proteins in the virulence of this bacterium. The results presented here compose the most comprehensive coverage of the exoproteome of a corynebacterial species so far