The open pan-genome architecture and virulence landscape of Mycobacterium bovis

Animal tuberculosis (TB) is an emergent disease caused by Mycobacterium bovis, one of the animal-adapted ecotypes of the Mycobacterium tuberculosis complex (MTC). In this work, whole-genome comparative analyses of 70 M. bovis were performed to gain insights into the pan-genome architecture. The comparison across M. bovis predicted genome composition enabled clustering into the core- and accessory-genome components, with 2736 CDS for the former, while the accessory moiety included 3897 CDS, of which 2656 are restricted to one/two genomes only. These analyses predicted an open pan-genome architecture, with an average of 32 CDS added by each genome and show the diversification of discrete M. bovis subpopulations supported by both core- and accessory-genome components. The functional annotation of the pan-genome classified each CDS into one or several COG (Clusters of Orthologous Groups) categories, revealing ‘transcription’ (total average CDSs, n=258), ‘lipid metabolism and transport’ (n=242), ‘energy production and conversion’ (n=214) and ‘unknown function’ (n=876) as the most represented. The closer analysis of polymorphisms in virulence-related genes in a restrict group of M. bovis from a multi-host system enabled the identification of clade-monomorphic non-synonymous SNPs, illustrating clade-specific virulence landscapes and correlating with disease severity. This first comparative pan-genome study of a diverse collection of M. bovis encompassing all clonal complexes indicates a high percentage of accessory genes and denotes an open, dynamic non-conservative pan-genome structure, with high evolutionary potential, defying the canons of MTC biology. Furthermore, it shows that M. bovis can shape its virulence repertoire, either by acquisition and loss of genes or by SNP-based diversification, likely towards host immune evasion, adaptation and persistence.info:eu-repo/semantics/publishedVersio

Genome-wide estimation of recombination, mutation and positive selection enlightens diversification drivers of Mycobacterium bovis

Genome sequencing has reinvigorated the infectious disease research feld, shedding light on disease epidemiology, pathogenesis, host–pathogen interactions and also evolutionary processes exerted upon pathogens. Mycobacterium tuberculosis complex (MTBC), enclosing M. bovis as one of its animal-adapted members causing tuberculosis (TB) in terrestrial mammals, is a paradigmatic model of bacterial evolution. As other MTBC members, M. bovis is postulated as a strictly clonal, slowly evolving pathogen, with apparently no signs of recombination or horizontal gene transfer. In this work, we applied comparative genomics to a whole genome sequence (WGS) dataset composed by 70 M. bovis from diferent lineages (European and African) to gain insights into the evolutionary forces that shape genetic diversifcation in M. bovis. Three distinct approaches were used to estimate signs of recombination. Globally, a small number of recombinant events was identifed and confrmed by two independent methods with solid support. Still, recombination reveals a weaker efect on M. bovis diversity compared with mutation (overall r/m= 0.037). The diferential r/m average values obtained across the clonal complexes of M. bovis in our dataset are consistent with the general notion that the extent of recombination may vary widely among lineages assigned to the same taxonomical species. Based on this work, recombination in M. bovis cannot be excluded and should thus be a topic of further efort in future comparative genomics studies for which WGS of large datasets from diferent epidemiological scenarios across the world is crucial. A smaller M. bovis dataset (n= 42) from a multi-host TB endemic scenario was then subjected to additional analyses, with the identifcation of more than 1,800 sites wherein at least one strain showed a single nucleotide polymorphism (SNP). The majority (87.1%) was located in coding regions, with the global ratio of non-synonymous upon synonymous alterations (dN/dS) exceeding 1.5, suggesting that positive selection is an important evolutionary force exerted upon M. bovis. A higher percentage of SNPs was detected in genes enriched into “lipid metabolism”, “cell wall and cell processes” and “intermediary metabolism and respiration” functional categories, revealing their underlying importance in M. bovis biology and evolution. A closer look on genes prone to horizontal gene transfer in the MTBC ancestor and included in the 3R (DNA repair, replication and recombination) system revealed a global average negative value for Taijima’s D neutrality test, suggesting that past selective sweeps and population expansion after a recent bottleneck remain as major evolutionary drivers of the obligatory pathogen M. bovis in its struggle with the host.info:eu-repo/semantics/publishedVersio

Preoperative assessment of deep myometrial and cervical invasion in endometrial carcinoma: comparison of magnetic resonance imaging and histopathologic evaluation

This study aimed to evaluate the accuracy of magnetic resonance imaging (MRI) in the detection of deep myometrial invasion and cervical extension by endometrial carcinoma. This prospective study included 101 patients with histologically documented endometrial carcinoma, between July 1998 and April 2004. The findings of preoperative pelvic MRI were compared with histological diagnosis. From 101 cases studied by pelvic MRI, 43 were classified as deep myometrial invasion (50% of myometrium), where the pathological evaluation confirmed as having deep myometrial invasion. Cervical extension in the MRI study was found in 19 cases. Pathologic study found cervical extension and/or invasion in 31 cases including all cases identified by MRI. The accuracy, sensitivity and specificity of MRI were 95%, 89%, 100%, detecting deep myometrial invasion and 88%, 61%, 100%, detecting cervical invasion, respectively. The high accuracy achieved makes MRI an adequate method for determine the depth of myometrial and cervical invasion in endometrial carcinoma

Modulating intermolecular interactions and rheological properties in silk fibroin using ultrasound frequencies

info:eu-repo/semantics/draf

Impact of macroeconomic indicators on bankruptcy prediction models: case of the Portuguese construction sector

The importance of macroeconomic indicators on the performance of bankruptcy prediction models has been a contentious issue, due in part to a lack of empirical evidence. Most indicators are primarily centered around a company's internal environment, overlooking the impact of the economic cycle on the status of the company. This research brings awareness about the combination of microeconomic and macroeconomic factors. To do this, a new model based on logistic regression was combined with principal component analysis to determine the indicators that best explained the variations in the dataset studied. The sample used comprised data from 1,832 Portuguese construction companies from 2009 to 2019. The empirical results demonstrated an average accuracy rate of 90% up until three years before the bankruptcy. The microeconomic indicators with statistical significance fell within the category of liquidity ratios, solvency and financial autonomy ratios. Regarding the macroeconomic indicators, the gross domestic product and birth rate of enterprises proved to increase the accuracy of bankruptcy prediction more than using only microeconomic factors. A practical implication of the results obtained is that construction companies, as well as investors, government agencies and banks, can use the suggested model as a decision-support system. Furthermore, consistent use can lead to an effective method of preventing bankruptcy by spotting early warning indicators

Inventory of attitudes toward seeking mental health services: psychometric properties among adolescents

Mental health is an essential element of wellness; however, some populations are more vulnerable to mental health problems. Suicide is the second-highest cause of death in young people and help-seeking unwillingness is a significant obstacle to mental health interventions. Young people are especially reluctant to seek help and negative attitudes stand out as help-seeking barriers, highlighting the importance of evaluating these constructs. The Inventory of Attitudes toward Seeking Mental Health Services (IASMHS) is a measure that evaluates help-seeking attitudes; nevertheless, it lacks applicability in Portuguese youth. This study aims to explore the psychometric characteristics of the IASMHS in 750 Portuguese adolescents (52.3% female, Mage = 14.67), through exploratory and confirmatory factor analysis, and reliability and validity analyses. Results suggest that a three-factor structure was adequate, those factors being help-seeking propensity, indifference to stigma, and psychological openness. The IASMHS was shortened to 20 items and revealed satisfactory properties regarding internal consistency, convergent/discriminant validity, and temporal stability. These findings validate the IASMHS as an adequate tool in the assessment of young people’s help-seeking attitudes.info:eu-repo/semantics/publishedVersio

Advanced treatment for arthritic diseases based on the capture and inactivation of interleukin-6 by biofunctionalized polymeric nanoparticles

Arthritic diseases, such as osteoarthritis and rheumatoid arthritis, are associated with synovium inflammation (synovitis). Several pro-inflammatory cytokines, especially tumor necrosis factor-α (TNFα) and interleukins (IL), are important mediators of inflammation and articular cartilage destruction, supporting a potential possibility of anticytokine therapy in these diseases. IL-6 is one of the key regulators of the inflammatory response. Thus, human monoclonal antibodies against IL-6 may prevent its action, and consequently reduce inflammation after intra-articular (IA) injection. Indeed, several clinical trials have already demonstrated positive outcomes over disease progression. Although these treatments are very attractive, they are associated with limited efficacy because of the rapid clearance of antibodies by the synovium. A solution to overcome this problem is using nanoparticles (NPs) as a substrate to protect and extend the action of the antibodies. Natural-derived polymers, like chitosan (Ch) and hyaluronic acid (HA), are biocompatible and biodegradable polysaccharides, being HA a natural component of the extracellular matrix of articular cartilage. Therefore, biodegradable polymeric NPs represent a good candidate for IA administration. In the present work we propose natural biodegradable polymeric NPs biofunctionalized with immobilized antibodies that selectively capture and inactivate the pro-inflammatory cytokine IL-6, reducing synovium inflammation. Ch-HA NPs were successfully prepared by polyelectrolyte complexation and further stabilized through carbodiimide chemistry (ethyl(dimethylaminopropyl) carbodiimide (EDC)/Nhydroxysuccinimide (NHS)). The particle size and zeta potential of the NPs were optimized. Stable NPs with 121.8 ± 2.4 of particle diameter, 0.11 ± 0.01 of polydispersity index and +25.12 ± 1.86 mV of zeta potential were produced with 0.25 mg/mL of initial polymers concentrations, at pH 5 and with 50/200 mM of EDC/NHS concentration. The anti-IL-6 antibody was immobilized at the surface of Ch-HA NPs. After determining the maximum antibody immobilization ability (7 µg/mL), the capacity to capture the recombinant IL-6 was evaluated. The efficacy was around 94-97%. Biological assays demonstrated not only the cytocompatibility of the produced NPs with human articular chondrocytes (hACs) (Fig 1) and human macrophages, but also the benefits of the capture and inactivation of IL-6 after stimulation with monocyte-derived macrophage conditioned medium. In conclusion, it is foreseeable that these NPs will overcome the limitations of the abovementioned treatments, since such NPs will increase the therapeutic efficacy due to their subcellular size, non-toxicity and high stability, being a promising approach for the local and sustained treatment of arthritic diseases.info:eu-repo/semantics/publishedVersio