Experimental evidence for limited in vivo virulence of Mycobacterium africanum

Funding Information: The authors thank the excellent support from the i3S scientific platforms, namely Animal Facility and Translational Cytometry. Funding. This work was supported by Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 ? Operational Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through Funda??o para a Ci?ncia e a Tecnologia, Minist?rio da Ci?ncia, Tecnologia e Inova??o in the framework of the project ?Institute for Research and Innovation in Health Sciences? (POCI-01-0145-FEDER-007274), and by grants FCT ? Aga Khan Development Network (ref 333197025), POCI-01-0145-FEDER-028955 (to MS), PTDC/BIA-MIC/30692/2017, and UID/Multi/04413/2013 (to DM and MV). BC and KF were funded by FCT Ph.D. scholarships SFRH/BD/114403/2016 and SFRH/BD/114405/2016, respectively. The Gulbenkian Foundation is acknowledged for a field work research grant to BC, Bolsas de apoio ? investiga??o para estudantes de doutoramento dos PALOP, Ref. P-146397. DM and MS were supported by FCT through Estimulo Individual ao Emprego Cient?fico. Publisher Copyright: © Copyright © 2019 Cá, Fonseca, Sousa, Maceiras, Machado, Sanca, Rabna, Rodrigues, Viveiros and Saraiva.Tuberculosis remains a public health problem and a main cause of death to humans. Both Mycobacterium tuberculosis and Mycobacterium africanum cause tuberculosis. In contrast to M. tuberculosis, which is geographically spread, M. africanum is restricted to West Africa. Differences have also been found in the growth rate and type of disease caused by M. africanum, globally suggesting an attenuation of this bacteria. In this study, we used the mouse model of infection to follow the dynamics of M. africanum infection in terms of bacterial burdens and tissue pathology, as well as the immune response triggered. Our findings support a lower virulence of M. africanum as compared to M. tuberculosis, including in mice lacking IFN-γ, a major protective cytokine in tuberculosis. Furthermore, the lung immune response triggered by M. africanum infection in wild-type animals was characterized by a discrete influx of leukocytes and a modest transcriptional upregulation of inflammatory mediators. Our findings contribute to elucidate the pathogenesis of M. africanum, supporting the hypothesis that this is an attenuated member of the tuberculosis-causing bacteria. Understanding the biology of M. africanum and how it interacts with the host to establish infection will have implications for our knowledge of TB and for the development of novel and better tools to control this devastating disease.publishersversionpublishe

Fibrose cística e a triagem neonatal

The clinical and diagnostic aspects of cystic fibrosis have been extensively reviewed, with an emphasis on neonatal screening. This systematic literature review involved a search for relevant contributions in the PubMed and SciELO databases. The first references to cystic fibrosis date to the Middle Ages. Cystic fibrosis is the most frequent autosomal recessive hereditary disease among Caucasians (1:2,000 to 3,500). More than 1,000 mutations lead to the disease, the most common being "F508, with 70% prevalence among Canadian, Northern European, and American Caucasians and 23 to 55% prevalence among Brazilians. The basic defect is in chloride ion secretion. Cystic fibrosis screening has long been controversial, and after almost three decades, there are few nationwide programs (most are regional or local). However, the U.S. Centers for Disease Control and Prevention (CDC) has concluded that screening for cystic fibrosis is justified. The lack of a specific screening test and the ethnic heterogeneity of the Brazilian population pose challenges for neonatal screening.Aspectos clínicos e diagnósticos da fibrose cística são revistos de modo abrangente, com ênfase na triagem neonatal. Esta revisão sistematizada da literatura envolveu busca de contribuições relevantes nos bancos de dados PubMed e SciELO. Referências sobre fibrose cística existem desde a Idade Média. É a doença hereditária autossômica recessiva mais freqüente em caucasianos (1:2.000 a 3.500). Mais de mil mutações levam à doença, a mais comum: "F508 (prevalência: 70% em caucasianos canadenses, americanos e norte-europeus; de 23 a 55% em brasileiros). O defeito básico ocorre na secreção do íon cloro. Sua triagem é assunto polêmico e apesar de estar disponível há quase três décadas, por meio de diferentes protocolos, poucos programas de abrangência nacional existem. Entretanto, o Centers for Disease Control and Prevention, dos Estados Unidos, afirma que o rastreamento neonatal para fibrose cística é justificado. A falta de um teste específico e a heterogeneidade étnica da população brasileira dificultam sua triagem neonatal

Fibrose cística e a triagem neonatal

The clinical and diagnostic aspects of cystic fibrosis have been extensively reviewed, with an emphasis on neonatal screening. This systematic literature review involved a search for relevant contributions in the PubMed and SciELO databases. The first references to cystic fibrosis date to the Middle Ages. Cystic fibrosis is the most frequent autosomal recessive hereditary disease among Caucasians (1:2,000 to 3,500). More than 1,000 mutations lead to the disease, the most common being ΔF508, with 70% prevalence among Canadian, Northern European, and American Caucasians and 23 to 55% prevalence among Brazilians. The basic defect is in chloride ion secretion. Cystic fibrosis screening has long been controversial, and after almost three decades, there are few nationwide programs (most are regional or local). However, the U.S. Centers for Disease Control and Prevention (CDC) has concluded that screening for cystic fibrosis is justified. The lack of a specific screening test and the ethnic heterogeneity of the Brazilian population pose challenges for neonatal screening244Aspectos clínicos e diagnósticos da fibrose cística são revistos de modo abrangente, com ênfase na triagem neonatal. Esta revisão sistematizada da literatura envolveu busca de contribuições relevantes nos bancos de dados PubMed e SciELO. Referências sobre fibrose cística existem desde a Idade Média. É a doença hereditária autossômica recessiva mais freqüente em caucasianos (1:2.000 a 3.500). Mais de mil mutações levam à doença, a mais comum: ΔF508 (prevalência: 70% em caucasianos canadenses, americanos e norte-europeus; de 23 a 55% em brasileiros). O defeito básico ocorre na secreção do íon cloro. Sua triagem é assunto polêmico e apesar de estar disponível há quase três décadas, por meio de diferentes protocolos, poucos programas de abrangência nacional existem. Entretanto, o Centers for Disease Control and Prevention, dos Estados Unidos, afirma que o rastreamento neonatal para fibrose cística é justificado. A falta de um teste específico e a heterogeneidade étnica da população brasileira dificultam sua triagem neonata

New 1,3-diarylureas linked by C-C Suzuki coupling to the methyl 3-aminothieno[3,2-b]pyridine-2-carboxylate moiety: synthesis and fluorescence studies in solution and in lipid membranes

New six fluorescent 1,3-diarylureas linked by C-C Suzuki coupling to the 6-position of the methyl 3-aminothieno[3,2-b]pyridine-2-carboxylate moiety were prepared by reaction of the amino groups on the ortho or meta positions relative to the C-C bond of the Suzuki coupling products, with different para-substituted arylisocyanates (H, OMe, CN), in high to excellent yields. The fluorescence properties of the 1,3-diarylureas in solution and in lipid membranes of egg-yolk phosphatidylcholine (Egg-PC), dipalmitoyl phosphatidylcholine (DPPC), dipalmitoyl phosphatidylglycerol (DPPG) or dioctadecyldimethylammonium bromide (DODAB), with or without cholesterol (Ch), were studied. The six 1,3-diarylureas have reasonable fluorescence quantum yields in several solvents (between 0.02 and 0.69) and present a moderately solvent sensitive emission, but are not fluorescent in alcohols and water. The compounds bearing the arylurea moiety in the meta position relative to the C-C bond, especially with the OMe and CN substituents, present the better solvatochromic properties. Incorporation of the six compounds in lipid membranes indicates that all the compounds are deeply located in the hydrophobic region of the lipid bilayers, feeling the transition between the rigid gel phase and fluid phases.To the Foundation for the Science and Technology (FCT, Portugal) for inancial support to the NMR portuguese network (PTNMR, Bruker Avance III 400-Univ. Minho). To the FCT and FEDER (European Fund for Regional Development)-COMPETE-QREN-EU for financial support to the Research Centres, CQ/UM [PEst-C/QUI/UI0686/2011 (FCOMP-01-0124-FEDER-022716)] and CFUM [PEst-C/FIS/UI0607/2011 (F-COMP-01-0124-FEDER-022711)], and to the research projects PTDC/QUI/81238/2006 (FCOMP-01-0124-FEDER-007467) (photophysical studies) and PTDC/QUI-QUI/111060/2009 (F-COMP-01-0124-FEDER-015603) (organic synthesis)

Mycobacterium tuberculosis Infection Up-Regulates Sialyl Lewis X Expression in the Lung Epithelium

Glycans display increasingly recognized roles in pathological contexts, however, their impact in the host-pathogen interplay in many infectious diseases remains largely unknown. This is the case for tuberculosis (TB), one of the ten most fatal diseases worldwide, caused by infection of the bacteria Mycobacterium tuberculosis. We have recently reported that perturbing the core-2 O -glycans biosynthetic pathway increases the host susceptibility to M. tuberculosis infection, by disrupting the neutrophil homeostasis and enhancing lung pathology. In the present study, we show an increased expression of the sialylated glycan structure Sialyl-Lewis X (SLeX) in the lung epithelium upon M. tuberculosis infection. This increase in SLeX glycan epitope is accompanied by an altered lung tissue transcriptomic signature, with up-regulation of genes codifying enzymes that are involved in the SLeX core-2 O -glycans biosynthetic pathway. This study provides novel insights into previously unappreciated molecular mechanisms involving glycosylation, which modulate the host response to M. tuberculosis infection, possibly contributing to shape TB disease outcome

Quinone oxidoreductase from Staphylococcus aureus

Funding Information: Helena Gaspar is acknowledged for the HPLC analyses and Bruno Victor for advice on modelling. F.M.S. and M.S.S. are recipients of fellowships by Fundação para a Ciência e a Tecnologia (PD/BD/128213/2016 and PD/BD/128202/2016, respectively, both within the scope of the PhD program Molecular Biosciences PD/00133/2012). A.B. is recipient of a fellowship by Fundação para a Ciência e a Tecnologia UI/BD/153052/2022. The work was funded by Fundação para a Ciência e a Tecnologia ( PTDC/BIA-BQM/2599/2021 to M.M.P). The project was further supported by UIDB/04046/2020 and UIDP/04046/2020 Centre grants from FCT , Portugal (to BioISI), by LISBOA-01-0145-FEDER-007660 cofunded by FEDER through COMPETE2020-POCI and by Fundação para a Ciência e a Tecnologia and by UIDB/04612/2020 and UIDP/04612/2020 research unit grants from FCT (to Mostmicro). The NMR spectrometers are part of the National NMR Network (PTNMR) and are supported by Infrastructure Project N° 022161 (co-financed by FEDER through COMPETE 2020, POCI, and PORL and FCT through PIDDAC). Funding Information: Helena Gaspar is acknowledged for the HPLC analyses and Bruno Victor for advice on modelling. F.M.S. and M.S.S. are recipients of fellowships by Fundação para a Ciência e a Tecnologia (PD/BD/128213/2016 and PD/BD/128202/2016, respectively, both within the scope of the PhD program Molecular Biosciences PD/00133/2012). A.B. is recipient of a fellowship by Fundação para a Ciência e a Tecnologia UI/BD/153052/2022. The work was funded by Fundação para a Ciência e a Tecnologia (PTDC/BIA-BQM/2599/2021 to M.M.P). The project was further supported by UIDB/04046/2020 and UIDP/04046/2020 Centre grants from FCT, Portugal (to BioISI), by LISBOA-01-0145-FEDER-007660 cofunded by FEDER through COMPETE2020-POCI and by Fundação para a Ciência e a Tecnologia and by UIDB/04612/2020 and UIDP/04612/2020 research unit grants from FCT (to Mostmicro). The NMR spectrometers are part of the National NMR Network (PTNMR) and are supported by Infrastructure Project N° 022161 (co-financed by FEDER through COMPETE 2020, POCI, and PORL and FCT through PIDDAC). Publisher Copyright: © 2022 The Author(s)Staphylococcus aureus is an opportunistic pathogen and one of the most frequent causes for community acquired and nosocomial bacterial infections. Even so, its energy metabolism is still under explored and its respiratory enzymes have been vastly overlooked. In this work, we unveil the dihydroorotate:quinone oxidoreductase (DHOQO) from S. aureus, the first example of a DHOQO from a Gram-positive organism. This protein was shown to be a FMN containing menaquinone reducing enzyme, presenting a Michaelis-Menten behaviour towards the two substrates, which was inhibited by Brequinar, Leflunomide, Lapachol, HQNO, Atovaquone and TFFA with different degrees of effectiveness. Deletion of the DHOQO coding gene (Δdhoqo) led to lower bacterial growth rates, and effected in cell morphology and metabolism, most importantly in the pyrimidine biosynthesis, here systematized for S. aureus MW2 for the first time. This work unveils the existence of a functional DHOQO in the respiratory chain of the pathogenic bacterium S. aureus, enlarging the understanding of its energy metabolism.publishersversionpublishe

2,4,5-Triaminopyrimidines as blue fluorescent probes for cell viability monitoring: synthesis, photophysical properties, and microscopy applications

Monitoring cell viability is critical in cell biology, pathology, and drug discovery. Most cell viability assays are cell-destructive, time-consuming, expensive, and/or hazardous. Herein, we present a series of newly synthesized 2,4,5-triaminopyrimidine derivatives able to discriminate between live and dead cells. To our knowledge, these compounds are the first fluorescent nucleobase analogues (FNAs) with cell viability monitoring potential. These new fluorescent molecules are synthesized using highly efficient and cost- effective methods and feature unprecedented photophysical properties (longer absorption and emission wavelengths, environment-sensitive emission, and unprecedented brightness within FNAs). Using a live– dead Saccharomyces cerevisiae cell and theoretical assays, the fluorescent 2,4,5-triaminopyrimidine derivatives were found to specifically accumulate inside dead cells by interacting with dsDNA grooves, thus paving the way for the emergence of novel and safe fluorescent cell viability markers emitting in the blue region. As the majority of commercially available viability dyes emit in the green to red region of the visible spectrum, these novel markers might be useful to meet the needs of blue markers for co-staining combinations

Avaliação da composição corporal de jogadores de futebol sub-20 da cidade de Uberlândia, Minas Gerais

Introduction: Football depends on several variables to be performed at its highest performance, among these variables, one of the most important is body composition, as it is related to the athlete's physical capacity and sports performance, which can influence the team's results. Objective: To evaluate the body composition of young soccer athletes, in an under-20 team. Methodology: Sixteen soccer players from a sub-20 team, aged 18 to 20 years, were evaluated. Weight and height were measured; in addition, they performed the electrical bioimpedance test to assess the parameters of body composition, mainly muscle mass and fat percentage. From the muscle mass, the muscle mass index (BMI) was calculated, dividing the total muscle mass by height in meters squared. Result: Most athletes were above the ideal fat percentage (n = 12), however, most individuals had values close to the ideal, which may have been a variation of the body composition assessment method itself. We observed that 4 individuals had an adequate percentage of fat. Regarding the IMM, the average values were within the normal range (10.9 ± 0.6 kg / m2), with only 4 athletes having little muscle mass. Conclusion: Almost all athletes had muscle mass within the normal range, while the majority had values above the adequate percentage of fat. These results show that future nutritional interventions should be carried out in these athletes, focusing mainly on reducing the percentage of fat.Trabalho de Conclusão de Curso (Graduação)Introdução: O futebol depende de diversas variáveis para ser realizado no seu mais alto rendimento, entre essas variáveis, uma das mais importantes é a composição corporal, pois está relacionada com a capacidade física e desempenho esportivo do atleta, podendo influenciar nos resultados da equipe. Objetivo: Avaliar a composição corporal de jovens atletas de futebol, em uma equipe sub-20. Metodologia: Foram avaliados 16 atletas de futebol de campo de uma equipe sub-20, com idade de 18 a 20 anos. Foram mensurados peso e estatura; além disso, fizeram o exame de bioimpedância elétrica para avaliar os parâmetros de composição corporal, principalmente massa muscular e percentual de gordura. A partir da massa muscular, foi calculado o índice de massa muscular (IMM), dividindo a massa muscular total pela altura em metros ao quadrado. Resultado: A maioria dos atletas estava acima do percentual de gordura ideal (n= 12), entretanto, a maior parte dos indivíduos apresentava valores próximos ao ideal, o que pode ter sido uma variação do próprio método de avaliação da composição corporal. Observamos que 4 indivíduos apresentavam percentual de gordura adequado. Em relação ao IMM, os valores médios estavam dentro da normalidade (10,9 ± 0,6 kg/m2 ), sendo que apenas 4 atletas apresentavam pouca massa muscular. Conclusão: Quase todos os atletas apresentaram massa muscular dentro da normalidade, enquanto que a maioria apresentou valores acima do adequado de percentual de gordura. Estes resultados mostram que devem ser realizadas futuras intervenções nutricionais nestes atletas, focando principalmente na redução do percentual de gordura

Factorial Structure of the Morningness-Eveningness-Stability-Scale (MESSi) and Sex and Age Invariance

Assessing morningness-eveningness preferences (chronotype), an individual characteristic that is mirrored in daily mental and physiological fluctuations, is crucial given their overarching influence in a variety of domains. The current work aimed to investigate the best factor structure of an instrument recently presented to asses this characteristic: the Morningness-Eveningness-Stability-Scale improved (MESSi). For the first time, the originally proposed three-factor structure was pitched against a uni- and a two-factor solution. Another novelty was to establish that the best-fitting model would be invariant in relation to sex and age, two variables that influence chronotype. A Confirmatory Factor Analyses on the data obtained from a sample of 2096 German adults (age: 18–76; M = 25.5, SD = 7.64) revealed that the originally proposed three-factor structure of the MESSi – Morning Affect, Eveningness, and Distinctness – was the only one to achieve acceptable fit indicators. Furthermore, each scale obtained good internal consistency. In order to assess age invariance, following the literature on development and chronotype, our sample was divided into three age groups: 18–21 years, 22–31 years, and 32 years or older. Full measurement invariance of the three-factor model was found for sex and age. Regarding differences between sexes, females did not differ significantly from males in Morning Affect, but scored significantly lower on Eveningness and higher on Distinctness; this last result has been consistent across validation studies of the MESSi. With respect to age differences, the oldest group scored lower on Eveningness and Distinctness in comparison with the other two age-groups; the intermediate group (age: 22–31) scored lower on Morning Affect when compared to both the younger and older age groups. Additionally, both Eveningness and Distinctness were negatively correlated with age. This latter relation has been consistently reported in other validation studies. Our results reinforce the idea that the MESSi assesses three different components of chronotype in a reliable manner and that this instrument can be used to explore sex and age differences

Structural and biophysical insights into the mode of covalent binding of rationally designed potent BMX inhibitors.

The bone marrow tyrosine kinase in chromosome X (BMX) is pursued as a drug target because of its role in various pathophysiological processes. We designed BMX covalent inhibitors with single-digit nanomolar potency with unexploited topological pharmacophore patterns. Importantly, we reveal the first X-ray crystal structure of covalently inhibited BMX at Cys496, which displays key interactions with Lys445, responsible for hampering ATP catalysis and the DFG-out-like motif, typical of an inactive conformation. Molecular dynamic simulations also showed this interaction for two ligand/BMX complexes. Kinome selectivity profiling showed that the most potent compound is the strongest binder, displays intracellular target engagement in BMX-transfected cells with two-digit nanomolar inhibitory potency, and leads to BMX degradation PC3 in cells. The new inhibitors displayed anti-proliferative effects in androgen-receptor positive prostate cancer cells that where further increased when combined with known inhibitors of related signaling pathways, such as PI3K, AKT and Androgen Receptor. We expect these findings to guide development of new selective BMX therapeutic approaches