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The notion of normal proof theory, and yet it has been somewhat neglected by the systems of equational logic. The intention here is then to show the normalization procedure for the equational logic of the Labelled Natural Deduction system (LND). With this we believe we are making a step towards filling a gap in the literature on equational logic. Besides presenting a normalization procedure for the LND equational fragment, we employ a new method to prove the normalization theorems for equational logic: proof transformation by rewriting, based on a algebraic calculus on the 'rewrite reasons'. [ABSTRACT FROM PUBLISHER]

Given the urgent need for novel methods to control the spread of multidrug-resistant microorganisms, this study presents a green synthesis approach to produce silver nanoparticles (AgNPs) using the bark extract from Anadenanthera colubrina (Vell.) Brenan var. colubrina. The methodology included obtaining the extract and characterizing the AgNPs, which revealed antimicrobial activity against MDR bacteria. A. colubrina species is valued in indigenous and traditional medicine for its medicinal properties. Herein, it was employed to synthesize AgNPs with effective antibacterial activity (MIC = 19.53-78.12 μM) against clinical isolates from the ESKAPEE group, known for causing high hospitalization costs and mortality rates. Despite its complexity, AgNP synthesis is an affordable method with minimal environmental impacts and risks. Plant-synthesized AgNPs possess unique characteristics that affect their biological activity and cytotoxicity. In this work, A. colubrina bark extract resulted in the synthesis of nanoparticles measuring 75.62 nm in diameter, with a polydispersity index of 0.17 and an average zeta potential of -29 mV, as well as low toxicity for human erythrocytes, with a CC 50 value in the range of 961 μM. This synthesis underscores its innovative potential owing to its low toxicity, suggesting applicability across several areas and paving the way for future research.

Background: The maintenance of skeletal muscle plasticity upon changes in the environment, nutrient supply, and exercise depends on regulatory mechanisms that couple structural and metabolic adaptations. The mechanisms that interconnect both processes at the transcriptional level remain underexplored. Nr2f6, a nuclear receptor, regulates metabolism and cell differentiation in peripheral tissues. However, its role in the skeletal muscle is still elusive. Here, we aimed to investigate the effects of Nr2f6 modulation on muscle biology in vivo and in vitro., Methods: Global RNA-seq was performed in Nr2f6 knockdown C2C12 myocytes (N = 4-5). Molecular and metabolic assays and proliferation experiments were performed using stable Nr2f6 knockdown and Nr2f6 overexpression C2C12 cell lines (N = 3-6). Nr2f6 content was evaluated in lipid overload models in vitro and in vivo (N = 3-6). In vivo experiments included Nr2f6 overexpression in mouse tibialis anterior muscle, followed by gene array transcriptomics and molecular assays (N = 4), ex vivo contractility experiments (N = 5), and histological analysis (N = 7). The conservation of Nr2f6 depletion effects was confirmed in primary skeletal muscle cells of humans and mice., Results: Nr2f6 knockdown upregulated genes associated with muscle differentiation, metabolism, and contraction, while cell cycle-related genes were downregulated. In human skeletal muscle cells, Nr2f6 knockdown significantly increased the expression of myosin heavy chain genes (two-fold to three-fold) and siRNA-mediated depletion of Nr2f6 increased maximal C2C12 myocyte's lipid oxidative capacity by 75% and protected against lipid-induced cell death. Nr2f6 content decreased by 40% in lipid-overloaded myotubes and by 50% in the skeletal muscle of mice fed a high-fat diet. Nr2f6 overexpression in mice resulted in an atrophic and hypoplastic state, characterized by a significant reduction in muscle mass (15%) and myofibre content (18%), followed by an impairment (50%) in force production. These functional phenotypes were accompanied by the establishment of an inflammation-like molecular signature and a decrease in the expression of genes involved in muscle contractility and oxidative metabolism, which was associated with the repression of the uncoupling protein 3 (20%) and PGC-1α (30%) promoters activity following Nr2f6 overexpression in vitro. Additionally, Nr2f6 regulated core components of the cell division machinery, effectively decoupling muscle cell proliferation from differentiation., Conclusions: Our findings reveal a novel role for Nr2f6 as a molecular transducer that plays a crucial role in maintaining the balance between skeletal muscle contractile function and oxidative capacity. These results have significant implications for the development of potential therapeutic strategies for metabolic diseases and myopathies., (© 2024 The Authors. Journal of Cachexia, Sarcopenia and Muscle published by Wiley Periodicals LLC.)

Low-pass whole genome sequencing (LP-WGS) has been applied as alternative method to detect copy number variants (CNVs) in the clinical setting. Compared with chromosomal microarray analysis (CMA), the sequencing-based approach provides a similar resolution of CNV detection at a lower cost. In this study, we assessed the efficiency and reliability of LP-WGS as a more affordable alternative to CMA. A total of 1363 patients with unexplained neurodevelopmental delay/intellectual disability, autism spectrum disorders, and/or multiple congenital anomalies were enrolled. Those patients were referred from 15 nonprofit organizations and university centers located in different states in Brazil. The analysis of LP-WGS at 1x coverage (>50kb) revealed a positive testing result in 22% of the cases (304/1363), in which 219 and 85 correspond to pathogenic/likely pathogenic (P/LP) CNVs and variants of uncertain significance (VUS), respectively. The 16% (219/1363) diagnostic yield observed in our cohort is comparable to the 15%-20% reported for CMA in the literature. The use of commercial software, as demonstrated in this study, simplifies the implementation of the test in clinical settings. Particularly for countries like Brazil, where the cost of CMA presents a substantial barrier to most of the population, LP-WGS emerges as a cost-effective alternative for investigating copy number changes in cytogenetics., (© 2024 Wiley Periodicals LLC.)

Vulvovaginal candidiasis (VVC) remains a prevalent fungal disease, characterized by challenges, such as increased fungal resistance, side effects of current treatments, and the rising prevalence of non- albicans Candida spp. naturally more resistant. This study aimed to propose a novel therapeutic approach by investigating the antifungal properties and toxicity of 2-hydroxychalcone (2-HC) and 3'-hydroxychalcone (3'-HC), both alone and in combination with fluconazole (FCZ) and clotrimazole (CTZ). A lipid carrier (LC) was also developed to deliver these molecules. The study evaluated in vitro anti- Candida activity against five Candida species and assessed cytotoxicity in the C33-A cell line. The safety and therapeutic efficacy of in vivo were tested using an alternative animal model, Galleria mellonella . The results showed antifungal activity of 2-HC and 3'-HC, ranging from 7.8 to 31.2 as fungistatic and 15.6 to 125.0 mg/L as fungicide effect, with cell viability above 80% from a concentration of 9.3 mg/L (2-HC). Synergistic and partially synergistic interactions of these chalcones with FCZ and CTZ demonstrated significant improvement in antifungal activity, with MIC values ranging from 0.06 to 62.5 mg/L. Some combinations reduced cytotoxicity, achieving 100% cell viability in many interactions. Additionally, two LCs with suitable properties for intravaginal application were developed. These formulations demonstrated promising therapeutic efficacy and low toxicity in Galleria mellonella assays. These results suggest the potential of this approach in developing new therapies for VVC.

Background: Immune tolerance induction (ITI) is the treatment of choice to eradicate neutralizing anti-factor (F)VIII alloantibodies (inhibitors) in people with inherited hemophilia A. However, it is not successful in 10% to 40% of the cases. The biological mechanisms and biomarkers associated with ITI outcome are largely unknown., Objectives: The aim of this study was to investigate the association of plasma cytokines (interferon-γ, tumor necrosis factor, interleukin [IL]-2, IL-4, IL-5, IL-6, IL-10, and IL-17A), chemokines (IL-8/CXCL8, RANTES/CCL5, MIG/CXCL9, MCP-1/CCL2, and IP-10/CXCL10), and anti-FVIII immunoglobulin (Ig) G total, IgG1, and IgG4 with ITI outcome., Methods: In this cross-sectional analysis of the Brazilian Immune Tolerance Study, we assessed plasma levels of anti-FVIII IgGs using an enzyme-linked immunosorbent assay with plasma-derived FVIII and recombinant FVIII as target antigens, immobilized in microplates., Results: We assayed 98 plasma samples of moderately severe and severe (FVIII activity, <2%) people with hemophilia A after completion of a first ITI course. Levels of anti-recombinant FVIII IgG total and IgG4 were higher in people with hemophilia A who failed ITI (IgG total optical density [OD], 0.37; IQR, 0.15-0.73; IgG4 OD, 2.19; IQR, 0.80-2.52) than in those who had partial (IgG total OD, 0.03; IQR, 0.00-0.14; IgG4 OD, 0.39; IQR, 0.09-1.11; P  < .0001 for both) or complete success (IgG total OD, 0.04; IQR, 0.00-0.07; IgG4 OD, 0.07; IQR, 0.06-0.40; P  < .0001 for both). Plasma cytokines, chemokines, and anti-FVIII IgG1 were not associated with ITI outcome., Conclusion: Our results show that high levels of plasma anti-FVIII IgG4 and IgG total are associated with ITI failure., (© 2024 The Authors.)

We report herein an enantioselective palladium-catalyzed Heck-Matsuda reaction for the desymmetrization of N -protected 2,5-dihydro-1 H -pyrroles with aryldiazonium salts, using the chiral N , N -ligand ( S )-PyraBox. This strategy has allowed straightforward access to a diversity of 4-aryl-γ-lactams via Heck arylation followed by a sequential Jones oxidation. The overall method displays a broad scope and good enantioselectivity, favoring the ( R ) enantiomer. The applicability of the protocol is highlighted by the efficient enantioselective syntheses of the selective phosphodiesterase-4-inhibitor rolipram and the commercial drug baclofen as hydrochloride., (Copyright © 2024, de Oliveira et al.)

Introduction: Diversifying voice therapy strategies is critical to get children and adolescents to adhere to the therapeutic process., Purpose: To investigate the face-to-face and distance playful strategies have been used for vocal health education and voice therapy in children and adolescents., Methods: An integrative literature review was carried out based on the recommendations of the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes (PRISMA). A search for full texts was conducted on the Virtual Health Library, Scientific Electronic Library Online (SCIELO) and PuBMed databases using the following descriptors, and their corresponding Medical Subject Headings (MeSH): voice (voz); voice quality (qualidade da voz); dysphonia (disfonia); voice disorders (distúrbios da fala); voice training (treinamento da voz); telemedicine (telemedicina); therapeutics (terapêutica); health promotion (promoção da saúde); group practice (prática de grupo); recreation therapy (terapia recreacional); child (criança); preschool (pré-escolar) and adolescent (adolescente)., Results: After full reading of the studies, 15 articles were included and categorized into the following topics: face-to-face and distance playful strategies for voice therapy in children; playful strategies with gamification for education and for voice therapy in children; and vocal health educational actions in children., Conclusion: The study found a higher number of studies that carried out educational actions and playful strategies, while fewer studies were related to the production of materials, gamification and online services for voice therapy in children. Given the proven benefits of this strategy for children, further studies focusing on the development and application of playful strategies, such as the use of gamification, should be conducted in order to stimulate the skills and adherence of this population to therapy., (Copyright © 2021 The Voice Foundation. Published by Elsevier Inc. All rights reserved.)

This study aimed to assess the activity of AgNPs biosynthesized by Fusarium oxysporum (bio-AgNPs) against multidrug-resistant uropathogenic Proteus mirabilis, and to assess the antibacterial activity of catheters coated with bio-AgNPs. Broth microdilution and time-kill kinetics assays were used to determine the antibacterial activity of bio-AgNPs. Catheters were coated with two (2C) and three (3C) bio-AgNPs layers using polydopamine as crosslinker. Catheters were challenged with urine inoculated with P. mirabilis to assess the anti-incrustation activity. MIC was found to be 62.5 µmol l-1, causing total loss of viability after 4 h and bio-AgNPs inhibited biofilm formation by 76.4%. Catheters 2C and 3C avoided incrustation for 13 and 20 days, respectively, and reduced biofilm formation by more than 98%, while the pristine catheter was encrusted on the first day. These results provide evidence for the use of bio-AgNPs as a potential alternative to combat of multidrug-resistant P. mirabilis infections., (© 2024. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The need to reconcile food production, the safeguarding of nature, and the protection of public health is imperative in a world of continuing global change, particularly in the context of risks of emerging zoonotic disease (EZD). In this paper, we explored potential land use strategies to reduce EZD risks using a landscape approach. We focused on strategies for cases where the dynamics of pathogen transmission among species were poorly known and the ideas of "land-use induced spillover" and "landscape immunity" could be used very broadly. We first modeled three different land-use change scenarios in a region of transition between the Cerrado and the Atlantic Forest biodiversity hotspots. The land-use strategies used to build our scenarios reflected different proportions of native vegetation cover, as a proxy of habitat availability. We then evaluated the effects of the proportion of native vegetation cover on the occupancy probability of a group of mammal species and analyzed how the different land-use scenarios might affect the distribution of species in the landscape and thus the risk of EZD. We demonstrate that these approaches can help identify potential future EZD risks, and can thus be used as decision-making tools by stakeholders, with direct implications for improving both environmental and socio-economic outcomes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Roque, Bellón, Guerra, Valente-Neto, Santos, Melo, Nobre Arcos, de Oliveira, Valle Nunes, de Araujo Martins, Souza, Herrera, Tavares, Almeida-Gomes, Pays, Renaud, Gomes Barrios, Yon, Bowsher, Sullivan, Johnson, Grelle and Ochoa-Quintero.)

Intracellular calcium dysregulation, oxidative stress, and mitochondrial dysfunction are some of the main pathway contributors towards disease progression in Duchenne muscular dystrophy (DMD). This study is aimed at investigating the effects of light emitting diode therapy (LEDT) and idebenone antioxidant treatment, applied alone or together in dystrophic primary muscle cells from mdx mice, the experimental model of DMD. Mdx primary muscle cells were submitted to LEDT and idebenone treatment and evaluated for cytotoxic effects and calcium and mitochondrial signaling pathways. LEDT and idebenone treatment showed no cytotoxic effects on the dystrophic muscle cells. Regarding the calcium pathways, after LEDT and idebenone treatment, a significant reduction in intracellular calcium content, calpain-1, calsequestrin, and sarcolipin levels, was observed. In addition, a significant reduction in oxidative stress level markers, such as H 2 O 2 , and 4-HNE levels, was observed. Regarding mitochondrial signaling pathways, a significant increase in oxidative capacity (by OCR and OXPHOS levels) was observed. In addition, the PGC-1α, SIRT-1, and PPARδ levels were significantly higher in the LEDT plus idebenone treated-dystrophic muscle cells. Together, the findings suggest that LEDT and idebenone treatment, alone or in conjunction, can modulate the calcium and mitochondrial signaling pathways, such as SLN, SERCA 1, and PGC-1α, contributing towards the improvement of the dystrophic phenotype in mdx muscle cells. In addition, data from the LEDT plus idebenone treatment showed slightly better results than those of each separate treatment in terms of SLN, OXPHOS, and SIRT-1., (© 2023. The Author(s), under exclusive licence to Cell Stress Society International.)

Homologous recombination deficiency (HRD) has become an important prognostic and predictive biomarker for patients with high-grade serous ovarian cancer who may benefit from poly-ADP ribose polymerase inhibitors (PARPi) and platinum-based therapies. HRD testing provides relevant information to personalize patients' treatment options and has been progressively incorporated into diagnostic laboratories. Here, we assessed the performance of an in-house HRD testing system deployable in a diagnostic clinical setting, comparing results from two commercially available next-generation sequencing (NGS)-based tumor tests (SOPHiA DDM TM HRD Solution and AmoyDx ® (HRD Focus Panel)) with the reference assay from Myriad MyChoice ® (CDx). A total of 85 ovarian cancer samples were subject to HRD testing. An overall strong correlation was observed across the three assays evaluated, regardless of the different underlying methods employed to assess genomic instability, with the highest pairwise correlation between Myriad and SOPHiA (R = 0.87, p -value = 3.39 × 10 -19 ). The comparison of the assigned HRD status to the reference Myriad's test revealed a positive predictive value (PPV) and negative predictive value (NPV) of 90.9% and 96.3% for SOPHiA's test, while AmoyDx's test achieved 75% PPV and 100% NPV. This is the largest HRD testing evaluation using different methodologies and provides a clear picture of the robustness of NGS-based tests currently offered in the market. Our data shows that the implementation of in-house HRD testing in diagnostic laboratories is technically feasible and can be reliably performed with commercial assays. Also, the turnaround time is compatible with clinical needs, making it an ideal alternative to offer to a broader number of patients while maintaining high-quality standards at more accessible price tiers.

Background: Coronavirus disease 2019 (COVID-19) is a viral infection caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Although respiratory manifestations have received greater visibility during the pandemic caused by this virus, numerous neurological complaints related to coronavirus 2 infection have been documented in several countries. These records suggest that this pathogen presents neurotropism, and it can cause different neurological conditions of varying intensity., Objective: To investigate the ability of coronavirus 2 to invade the central nervous system (CNS) and its neurological clinical outcomes., Methods: The present study consists in a comprehensive literature review of the records available in the PubMed, SciELO, and Google Scholar databases. The descriptors COVID-19 , brain and physiopathology , associated with the Boolean operator AND , were used in the search. Regarding the inclusion and exclusion criteria, we selected the papers published since 2020 with the highest number of citations., Results: We selected 41 articles, most of them in English. The main clinical manifestation associated with COVID-19 patients was headache, but cases of anosmia, hyposmia, Guillain-Barré syndrome, and encephalopathies were also described with considerable frequency., Conclusion: Coronavirus-2 presents neurotropism, and it can reach the CNS by hematogenous dissemination and by direct infection of the nerve endings. It causes brain injuries through several mechanisms, such as cytokine storm, microglial activation, and an increase in thrombotic factors., Competing Interests: The authors have no conflict of interest to declare., (Academia Brasileira de Neurologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution 4.0 International License, permitting copying and reproduction so long as the original work is given appropriate credit (https://creativecommons.org/licenses/by/4.0/).)

The objective of this research was to evaluate the thermal exchanges, physiological responses, productive performance and carcass yield of Guinea Fowl confined under thermoneutral conditions and under thermal stress. For the experiment, 96 animals were confined in 8 experimental boxes of 1 m 2 of area, each, divided in equal numbers and placed inside two distinct climatic chambers, where the birds were distributed in a completely randomized design, with two treatments (air temperatures of 26 and 32 °C, respectively). For the collection of physiological responses and carcass yield 16 birds were evaluated and for the collection of data on feed and water consumption and productive responses, 48 birds per treatment were evaluated. The environmental variables (air temperature (AT), air relative humidity and wind speed), temperature and humidity index (THI), heat exchanges, physiological responses (respiratory rate, surface temperature, cloacal temperature and eyeball temperature), feed (FC) and water (WC) consumption and production responses (weight gain, feed conversion index and carcass yield) of the birds were evaluated. With the elevation of the AT, it could be noticed that the THI went from a thermal comfort condition to an emergency condition, where the birds lost part of their feathers, increased all physiological responses evaluated, and consequently, reduced by 53.5% the amount of heat dissipated in the sensible form and increased by 82.7% the heat losses in the latent form, increasing also the WC. ATs of up to 32 °C did not significantly affect the productive performance and carcass yield of the guinea fowl., (© 2023. The Author(s) under exclusive licence to International Society of Biometeorology.)

The first objective of this study was to develop a thermal stress index for sheep based on environmental and animal data collected in a climate chamber under various environmental conditions. The second objective was to compare published indices of thermal comfort and the proposed index, testing them with the data from this study, with the objective of pointing out the most adequate index to be used by breeders when choosing management procedures. A total of 3024 data were obtained for the physiological variables of the sheep exposed to the eight thermal conditions in the climatic chamber and in ambient condition, kept three days in each thermal condition. A principal component analysis summarized the measurements of physiological variables into only one variable (y1). Using SigmaPlot software, multiple regression of y1 with the environmental variables and their combinations produced a number of indices. The equation chosen was the heat stress index for sheep, TSI = 24.153 - (0.0523*A T ) + (0.746*B GT ) + (4.104*Vp), with R 2  = 0.668. The correlations presented high values, where these correlation values were assumed to indicate the efficiency of each index as indicators of the animals' response to the environment. Thus, it was assumed that the TSI presents a high efficiency., Competing Interests: Declaration of competing interest The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the article., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Resistant bacteria may kill more people than COVID-19, so the development of new antibacterials is essential, especially against microbial biofilms that are reservoirs of resistant cells. Silver nanoparticles (bioAgNP), biogenically synthesized using Fusarium oxysporum , combined with oregano derivatives, present a strategic antibacterial mechanism and prevent the emergence of resistance against planktonic microorganisms. Antibiofilm activity of four binary combinations was tested against enteroaggregative Escherichia coli (EAEC) and Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC): oregano essential oil (OEO) plus bioAgNP, carvacrol (Car) plus bioAgNP, thymol (Thy) plus bioAgNP, and Car plus Thy. The antibiofilm effect was accessed using crystal violet, MTT, scanning electron microscopy, and Chromobacterium violaceum anti-quorum-sensing assays. All binary combinations acted against preformed biofilm and prevented its formation; they showed improved antibiofilm activity compared to antimicrobials individually by reducing sessile minimal inhibitory concentration up to 87.5% or further decreasing biofilm metabolic activity and total biomass. Thy plus bioAgNP extensively inhibited the growth of biofilm in polystyrene and glass surfaces, disrupted three-dimensional biofilm structure, and quorum-sensing inhibition may be involved in its antibiofilm activity. For the first time, it is shown that bioAgNP combined with oregano has antibiofilm effect against bacteria for which antimicrobials are urgently needed, such as KPC.

A systematic review and network meta-analysis was performed to provide evidence for the best polishing protocol for different types of resin composites to minimize surface roughness. A search was performed in PubMed, Scopus, Web of Science, LILACS, BBO, EMBASE, and Cochrane Library on July 2, 2019 (updated in December, 2020). In vitro studies that included at least two systems for polishing resin composites and analyzed surface roughness were included. The risk of bias was evaluated. A random-effects Bayesian-mixed treatment comparison model was used to compare surface roughness in resin composites with the different types of polishers. Surface under the cumulative ranking curve (SUCRA) analysis was performed to rank the probability for the best polishing system. After removal of duplicates, title and abstract screening yielded 34 studies. Network meta-analysis was not possible for hybrid and microhybrid composites. SUCRA analysis showed that abrasive paper discs allowed greater surface smoothness for nanohybrid and nanofill composites to a probability of between 83% and 91.6%. Silicon carbide brush had a 78.2% probability of being the best system for microfill composites. The use of abrasive paper disc polishers showed a favorable result in nanofill and nanohybrid resin composites. Silicon carbide brush has a greater chance of promoting a smoother surface for microfill resin composites., (© 2023 Scandinavian Division of the International Association for Dental Research. Published by John Wiley & Sons Ltd.)

Background: In front of the physical and social isolation, as well as feelings of fear and instability imposed by the pandemic of COVID-19, especially for risk groups such as people 50 + , it became even more relevant to discuss the formulation of personal plans for this age population. This study aimed to analyse the individual plans of people 50 + , considering their perception, feelings and life experiences during the COVID-19 pandemic., Methods: This is a mixed study (quali-quantitative), using Minayo's methodology for content analysis and frequency analyses, made with 39 participants from the University of Brasília Educational Program, Universidade do Envelhecer - UniSER/UnB. We used a word cloud system and a wheel of life tool to showcase the results., Results: Analysing professional achievements and situations participants want to overcome, the categories of feelings that stand out were Loving Relationships, Sadness, Family Absence, Grief, Trauma and Regret. Intellectual Development, Personal Fulfilment, Purpose and Creativity, Hobbies and Fun were the most mentioned personal plans displayed by the wheel of life. The key personal changes mentioned were to be less shy, prioritise themselves, change how they interact with others, and focus on their health., Conclusions: This study concludes that perception, feelings and life experiences during the COVID-19 pandemic did not seem to directly affect the path in elaborating personal plans., (© 2023. The Author(s).)

Aim: To investigate the anti-inflammatory effects of the sesquiterpenes α-humulene and β-caryophyllene on pterygium fibroblasts., Methods: Primary cultures of pterygium fibroblasts were established. Third passage pterygium fibroblasts were exposed to α-humulene and β-caryophyllene separately and together. The cell viability was assessed by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) assay at 12, 24, 48, and 72h after exposure. The levels of the inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor (TNF)-α and IL-10 in the conditioned culture medium were assessed by enzyme-linked immunosorbent assay (ELISA) at 12, 24 and 48h after exposure. Data were statistically analyzed using Friedman repeated measures analysis of variances on ranks., Results: The 25 µmol/L β-caryophyllene induced significant decrease in the IL-6 production by pterygium fibroblasts 48h after the exposure ( P =0.041). The levels of IL-1β, IL-8, IL-10, and TNF-α were very low and had no statistically significant variations after exposure to α-humulene, β-caryophyllene, or both compounds together., Conclusion: The exposure to 25 µmol/L of β-caryophyllene significantly reduce the production of IL-6 by pterygium fibroblasts after 48h. This sesquiterpene may be a potential alternative adjuvant agent for the treatment of pterygium., (International Journal of Ophthalmology Press.)

Background: Immune tolerance induction (ITI) is the treatment of choice for eradication of anti-factor VIII (FVIII) neutralizing alloantibodies (inhibitors) in people with inherited hemophilia A and high-responding inhibitor (PwHA-HRi). The association between ITI outcome and time elapsed between inhibitor detection and start of ITI (∆t inhi-ITI ) is debatable., Objective: The aim of this study was to evaluate this association among a large cohort of severe PwHA-HRi., Methods: Severe (factor VIII activity level <1%) PwHA-HRi on ITI (n = 142) were enrolled in 15 hemophilia treatment centers. PwHA-HRi were treated according to the Brazilian ITI Protocol. ITI outcomes were defined as success (i.e., recovered responsiveness to exogenous FVIII) and failure (i.e., no responsiveness to exogenous FVIII and requirement of bypassing agents to control bleeding)., Results: Median ages at inhibitor detection and at ITI start were 3.2 years (interquartile range [IQR], 1.6-8.1) and 6.9 years [IQR, 2.6-20.1), respectively. PwHA-HRi were stratified according to ∆t inhi-ITI quartiles: first (0.0-0.6 year), second (>0.6-1.7 year), third (>1.7-9.2 years), and fourth quartile (>9.2-24.5 years). The overall success rate was 65.5% (93/142), with no difference among first, second, third, and fourth quartiles (62.9%, 69.4%, 58.3%, and 71.4%, respectively) even after adjusting the analyses for potential confounders., Conclusion: In conclusion, delayed ITI start is not associated with failure of ITI in PwHA-HRi. Therefore, ITI should be offered for these patients, regardless of the time elapsed between the detection of inhibitor and the ITI start., (© 2022 International Society on Thrombosis and Haemostasis.)

The present study demonstrates the biocontrol potential of a plant growth-promoting bacterial strain using three different approaches: (i) an in vitro evaluation of antagonistic activity against important phytopathogenic fungi; (ii) an evaluation under greenhouse conditions with strawberry plants to assess the control of gray mold; and (iii) an in silico whole genome sequence mining to assign genetic features such as gene clusters or isolated genes to the strain activity. The in vitro assay showed that the B.BV10 strain presented antagonistic activity, inhibiting the mycelial growth in all the phytopathogenic fungi evaluated. The application of the Bacillus velezensis strain B.BV10 under greenhouse conditions reduced the presence of Botrytis cinerea and increased the mean fruit biomass. The genome of B.BV10 was estimated at 3,917,533 bp, with a GC content of 46.6% and 4088 coding DNA sequences, and was identified as B. velezensis . Biosynthetic gene clusters related to the synthesis of the molecules with antifungal activity were found in its genome. Genes related to the regulation/formation of biofilms, motility, and the important properties for the rhizospheric colonization were also found in the genome. The current study offers a comprehensive understanding of the genomic architecture and control activity of phytopathogenic fungi by the B. velezensis strain B.BV10 that may substantiate the industrialization of this strain in the future.

The development of bio-based products has increased in recent years, and species of the Bacillus genus have been widely used for product development due to their elevated production of antimicrobial molecules and resistance to extreme environmental conditions through endospore formation. In this context, the antifungal potential of Bacillus velezensis CMRP 4489 was investigated using in silico predictions of secondary metabolites in its genome and in vitro tests against the following phytopathogenic fungi: Sclerotinia sclerotiorum, Macrophomina phaseolina, and Botrytis cinerea. The in-silico predictions indicated that CMRP 4489 possesses several Biosynthetic Gene Clusters (BGCs) capable of producing molecules with antifungal properties and other non-identified BGCs. The in vitro assay results evidenced strong antifungal activity, inhibiting more than 60% of the tested fungi, and the isolate's molecules were stable under diverse physicochemical conditions. The in vitro assay evidenced significant antifungal activity, deformation of the hyphal structure in SS, biofilm formation capacity, and swarming motility. In the colonization assay, we observed attachment, colonization, and net-shaped biofilm formation, with the strain transitioning from the seeds to nearby structures. Therefore, CMRP 4489 showed to be a potential biocontrol agent against various diseases with agronomic importance and can be used under adverse environmental conditions., (© 2022. The Author(s).)

In this study, we evaluated the antimicrobial susceptibility, the presence of gene-encoding virulence factors and CRISPR systems, as well as the ability to produce lytic enzymes among clinical E. faecalis and E. faecium isolates (n = 44). All enterococci isolates showed phenotypes of multidrug resistance. E. faecalis and E. faecium isolates exhibited high-level aminoglycoside resistance phenotype, several of them harboring the aac(6')Ie-aph(2″)Ia and aph(3')-IIIa genes. The gene vanA was the most frequent among vancomycin-resistant E. faecium. High prevalence of the virulence genes esp and efaA were observed; hyl gene was more associated with E. faecium, while ace and efaA genes were more frequently detected in E. faecalis. Caseinase activity was frequently detected among the isolates. Gelatinase and DNAse activities predominated among E. faecalis, while hemolytic capability was frequent among E. faecium isolates. Twenty-nine isolates showed at least one CRISPR system investigated. Several enterococci isolates harbored the aac(6')-Ie-aph(2″)-Ia or aph(3')-IIIa genes and a CRISPR loci. CRISPR loci were positively correlated to efaA and gelE genes, and gelatinase and DNAse activities, while CRISPR loci absence was related to hyl gene presence. These results show that clinical isolates of E. faecalis and E. faecium harboring virulence genes show the concomitant presence of CRISPR loci and antibiotic resistance determinants., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Aline Dias Paiva reports financial support was provided by National Council for Scientific and Technological Development. Isabela Sguilla Rotta reports financial support was provided by Minas Gerais State Foundation of Support to the Research., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Multidrug-resistant bacteria have become a public health problem worldwide, reducing treatment options against several pathogens. If we do not act against this problem, it is estimated that by 2050 superbugs will kill more people than the current COVID-19 pandemic. Among solutions to combat antibacterial resistance, there is increasing demand for new antimicrobials. The antibacterial activity of binary combinations containing bioAgNP (biogenically synthesized silver nanoparticles using Fusarium oxysporum ), oregano essential oil (OEO), carvacrol (Car), and thymol (Thy) was evaluated: OEO plus bioAgNP, Car plus bioAgNP, Thy plus bioAgNP, and Car plus Thy. This study shows that the mechanism of action of Thy, bioAgNP, and Thy plus bioAgNP involves damaging the membrane and cell wall (surface blebbing and disruption seen with an electron microscope), causing cytoplasmic molecule leakage (ATP, DNA, RNA, and total proteins) and oxidative stress by enhancing intracellular reactive oxygen species and lipid peroxidation; a similar mechanism happens for OEO and Car, except for oxidative stress. The combination containing bioAgNP and oregano derivatives, especially thymol, shows strategic antibacterial mechanism; thymol disturbs the selective permeability of the cell membrane and consequently facilitates access of the nanoparticles to bacterial cytoplasm. BioAgNP-treated Escherichia coli developed resistance to nanosilver after 12 days of daily exposition. The combination of Thy and bioAgNP prevented the emergence of resistance to both antimicrobials; therefore, mixture of antimicrobials is a strategy to extend their life. For antimicrobials alone, minimal bactericidal concentration ranges were 0.3-2.38 mg/ml (OEO), 0.31-1.22 mg/ml (Car), 0.25-1 mg/ml (Thy), and 15.75-31.5 μg/ml (bioAgNP). The time-kill assays showed that the oregano derivatives acted very fast (at least 10 s), while the bioAgNP took at least 30 min to kill Gram-negative bacteria and 7 h to kill methicillin-resistant Staphylococcus aureus (MRSA). All the combinations resulted in additive antibacterial effect, reducing significantly minimal inhibitory concentration and acting faster than the bioAgNP alone; they also showed no cytotoxicity. This study describes for the first time the effect of Car and Thy combined with bioAgNP (produced with F. oxysporum components) against bacteria for which efficient antimicrobials are urgently needed, such as carbapenem-resistant strains ( E. coli , Klebsiella pneumoniae , Acinetobacter baumannii , and Pseudomonas aeruginosa ) and MRSA., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Scandorieiro, Rodrigues, Nishio, Panagio, de Oliveira, Durán, Nakazato and Kobayashi.)

Brevibacillus brevis LABIM17 is a bacterial isolate with biotechnological potential. Its draft genome sequence contains a chromosome of 5,950,202 bp, with 5,477 coding sequences, and exhibits 12 clusters involved in the production of secondary metabolites, which are likely responsible for its antimicrobial activity against several human and plant pathogens.

Rationale: The FDA-approved Dimethyl Fumarate (DMF) as an oral drug for Multiple Sclerosis (MS) treatment based on its immunomodulatory activities. However, it also caused severe adverse effects mainly related to the gastrointestinal system., Objective: Investigated the potential effects of solid lipid nanoparticles (SLNs) containing DMF, administered by inhalation on the clinical signs, central nervous system (CNS) inflammatory response, and lung function changes in mice with experimental autoimmune encephalomyelitis (EAE)., Materials and Methods: EAE was induced using MOG35-55 peptide in female C57BL/6J mice and the mice were treated via inhalation with DMF-encapsulated SLN (CTRL/SLN/DMF and EAE/SLN/DMF), empty SLN (CTRL/SLN and EAE/SLN), or saline solution (CTRL/saline and EAE/saline), every 72 h during 21 days., Results: After 21 days post-induction, EAE mice treated with DMF-loaded SLN, when compared with EAE/saline and EAE/SLN, showed decreased clinical score and weight loss, reduction in brain and spinal cord injury and inflammation, also related to the increased influx of Foxp3+ cells into the spinal cord and lung tissues. Moreover, our data revealed that EAE mice showed signs of respiratory disease, marked by increased vascular permeability, leukocyte influx, production of TNF-α and IL-17, perivascular and peribronchial inflammation, with pulmonary mechanical dysfunction associated with loss of respiratory volumes and elasticity, which DMF-encapsulated reverted in SLN nebulization., Conclusion: Our study suggests that inhalation of DMF-encapsulated SLN is an effective therapeutic protocol that reduces not only the CNS inflammatory process and disability progression, characteristic of EAE disease, but also protects mice from lung inflammation and pulmonary dysfunction., (© 2022 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.)

Cellulose nanofibers (CNF) were employed as the nanoreinforcement of a retrograded starch/pectin (RS/P) excipient to optimize its colon-specific properties. Although starch retrogradation ranged from 32 to 73%, CNF addition discretely disfavored the RS yield. This result agrees with the finding that in situ CNF reduces the presence of the RS crystallinity pattern. A thermal analysis revealed that the contribution of pectin improves the thermal stability of the RS/CNF mixture. Through a complete factorial design, it was possible to optimize the spray-drying conditions to obtain powders with high yield (57%) and low moisture content (1.2%). The powders observed by Field Emission Gum Scanning Electron Microscopy (FEG-SEM) had 1-10 µm and a circular shape. The developed methodology allowed us to obtain 5-aminosalicilic acid-loaded microparticles with high encapsulation efficiency (16-98%) and drug loading (1.97-26.63%). The presence of CNF in RS/P samples was responsible for decreasing the burst effect of release in simulated gastric and duodenal media, allowing the greatest mass of drug to be targeted to the colon. Considering that spray-drying is a scalable process, widely used by the pharmaceutical industry, the results obtained indicate the potential of these microparticles as raw material for obtaining other dosage forms to deliver 5-ASA to the distal parts of gastrointestinal tract, affected by inflammatory bowel disease.

Background: The Infant mortality rate indicates the quality of life of a population. Infant mortality has two important components: neonatal mortality, divided into early and late and post-neonatal mortality. The more developed a country is and the greater its population's well-being, the greater the weight of the neonatal component on infant mortality. In addition several factors may determine or be associated with the occurrence of infant deaths including maternal age. The teenage pregnancy rates in Latin America and the Caribbean remain the second highest in the world, In Brazil, between 2010 and 2015, for every thousand adolescents between 15 and 19 years old, about 69 became pregnant and gave birth to their babies. Thus, the objective of this study is to evaluate the trend of Early Neonatal Mortality Rates in children of pregnant adolescents, which occurred in the period 1996-2017, in the state of São Paulo, Brazil, according to the maternal age group., Methods: This is an ecological study of time series using official mortality data obtained from the Mortality Information System and live birth data obtained from the Live Birth Information System. Deaths of newborns aged between zero and six complete days were collected by place of residence. The trends in rates per 1,000 live births were calculated by Prais-Winsten regression, obtaining their annual percentage change (VPA) and the respective 95% confidence intervals, analyzed by age group. All analyzes were processed using the STATA 15.1 software., Results: In the state of São Paulo, between 1996 and 2017, 16,161 deaths were reported in children from zero to six days old and 2,320,584 live births in mothers aged 10-19 years, living in the state of São Paulo, Brazil. Of this total, it was observed that the early neonatal mortality rate decreased until the year 2005-2006, remained stationary after, and was higher in newborns of mothers aged 10-14 years (13.18 per 1,000) compared to mothers between 15-19 years (6.75 per 1,000)., Conclusions: In conclusion, although the early neonatal mortality rate showed a significant decreasing trend until approximately 2005, it remained stables after that., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at: http://dx.doi.org/10.21037/tp-20-438). The authors have no conflicts of interest to declare., (2021 Translational Pediatrics. All rights reserved.)

Oligonucleotide gene therapy (OGT) agents suppress specific mRNAs in cells and thus reduce the expression of targeted genes. The ability to unambiguously distinguish cancer from healthy cells can solve the low selectivity problem of OGT agents. Cancer RNA markers are expressed in both healthy and cancer cells with a higher expression level in cancer cells. We have designed a DNA-based construct, named DNA thresholder (DTh) that cleaves targeted RNA only at high concentrations of cancer marker RNA and demonstrates low cleavage activity at low marker concentrations. The RNA-cleaving activity can be adjusted within one order of magnitude of the cancer marker RNA concentration by simply redesigning DTh. Importantly, DTh recognizes cancer marker RNA, while cleaving targeted RNA; this offers a possibility to suppress vital genes exclusively in cancer cells, thus triggering their death. DTh is a prototype of computation-inspired molecular device for controlling gene expression and cancer treatment., (© 2021 Wiley-VCH GmbH.)

Background: Stingless bee honey has great therapeutic potential, especially as an antimicrobial agent. In the present study, we evaluated the in vitro antibacterial potential of honey from Melipona spp. with occurrence in Rio Branco-AC and Xapuri-AC from the Amazonian region. Samples were collected from the species Melipona eburnea, Melipona grandis, Melipona flavolineata and Melipona seminigra. The antibacterial activity of the honey samples was tested against standard Gram-positive and Gram-negative bacteria and two strains isolated from bovine mastitis., Results: In the agar diffusion assay, we observed antibacterial activity for the four honeys against the tested strains. The honey from M. flavolineata showed a minimmum inhibitory concentration (MIC) lower than 3.12% (v/v). The minimum bactericidal concentration values were larger than the MIC for most of the microorganisms tested. Scanning electron microscopy (SEM) showed the damaging effect of the honey of M. flavolineata on Staphylococcus aureus cells, as well as its inhibitory effect on cell division., Conclusion: The results of the present study demonstrate that the honey from stingless bees possesses in vitro antimicrobial activity against pathogenic bacteria. The effects observed by SEM show that honey from the Amazonian stingless bee M. flavolineata has promising therapeutic potential as a future antimicrobial agent. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

The main objective of this study was to evaluate Bacillus velezensis strain CMRP 4490 regarding its ability to inhibit soil-borne plant pathogens and to increase plant growth. The study included evaluation of in vitro antifungal control, sequencing the bacterial genome, mining genes responsible for the synthesis of secondary metabolites, root colonization ability, and greenhouse studies for the assessment of plant growth-promoting ability. The strain was obtained from soil samples in the north of Paraná in Brazil and was classified as a B. velezensis , which is considered a promising biological control agent. In vitro assay showed that B. velezensis CMRP 4490 presented antagonistic activity against Sclerotinia sclerotiorum , Macrophomina phaseolina , Botrytis cinerea , and Rhizoctonia solani with a mycelial growth inhibition of approximately 60%, without any significant difference among them. To well understand this strain and to validate its effect on growth-promoting rhizobacteria, it was decided to explore its genetic content through genome sequencing, in vitro , and greenhouse studies. The genome of CMRP 4490 was estimated at 3,996,396 bp with a GC content of 46.4% and presents 4,042 coding DNA sequences. Biosynthetic gene clusters related to the synthesis of molecules with antifungal activity were found in the genome. Genes linked to the regulation/formation of biofilms, motility, and important properties for rhizospheric colonization were also found in the genome. Application of CMRP 4490 as a coating film on soybean increased from 55.5 to 64% on germination rates when compared to the control; no differences were observed among treatments for the maize germination. The results indicated that B. velezensis CMRP 4490 could be a potential biocontrol agent with plant growth-promoting ability., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Teixeira, Mosela, Nicoletto, Ribeiro, Hungria, Youssef, Higashi, Mian, Ferreira, Gonçalves, Pereira and de Oliveira.)

Rotavirus (RV) is considered a major cause of acute viral gastroenteritis in young animals. RV is classified into nine species, five of which have been identified in pigs. Most studies worldwide have highlighted diarrhoea outbreaks caused by RVA, which is considered the most important RV species. In the present study, we described the detection and characterization of porcine RVB as a primary causative agent of diarrhoea outbreaks in pig herds in Brazil. The study showed a high frequency (64/90; 71.1%) of RVB diagnosis in newborn piglets associated with marked histopathological lesions in the small intestines. Phylogenetic analysis of the VP7 gene of wild-type RVB strains revealed a high diversity of G genotypes circulating in one geographic region of Brazil. Our findings suggest that RVB may be considered an important primary enteric pathogen in piglets and should be included in the routine differential diagnosis of enteric diseases in piglets.

The nuclear receptor PPARγ is essential to maintain whole-body glucose homeostasis and insulin sensitivity, acting as a master regulator of adipogenesis, lipid, and glucose metabolism. Its activation through natural or synthetic ligands induces the recruitment of coactivators, leading to transcription of target genes such as cytokines and hormones. More recently, post translational modifications, such as PPARγ phosphorylation at Ser273 by CDK5 in adipose tissue, have been linked to insulin resistance trough the dysregulation of expression of a specific subset of genes. Here, we investigate how this phosphorylation may disturb the interaction between PPARγ and some coregulator proteins as a new mechanism that may leads to insulin resistance. Through cellular and in vitro assays, we show that PPARγ phosphorylation inhibition increased the activation of the receptor, therefore the increased recruitment of PGC1-α and TIF2 coactivators, whilst decreases the interaction with SMRT and NCoR corepressors. Moreover, our results show a shift in the coregulators interaction domains preferences, suggesting additional interaction interfaces formed between the phosphorylated PPARγ and some coregulator proteins. Also, we observed that the CDK5 presence disturb the PPARγ-coregulator's synergy, decreasing interaction with PGC1-α, TIF2, and NCoR, but increasing coupling of SMRT. Finally, we conclude that the insulin resistance provoked by PPARγ phosphorylation is linked to a differential coregulators recruitment, which may promote dysregulation in gene expression., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Dias, Batista, Tittanegro, de Oliveira, Le Maire, Torres, Filho, Silveira and Figueira.)

The state of Mato Grosso do Sul, Brazil is an important region of South America regarding leishmaniasis, with the great diversity of sandflies reported since 1938 and wide biodiversity represented by its biomes (Atlantic Forest, Cerrado, and Pantanal). A checklist of sandflies is presented here for the state and respective biomes. The data base was compiled from primary and secondary data. The primary data collection involved automatic light traps at Miranda-Abobral Pantanal subregion and in subregion Baixa Nhecolância of Pantanal. The secondary data were obtained from entomology scientific collections and a literature review of articles from 1938 to 2019, including entomological museum collections. A total of 71 species were reported, belonging to 14 genera and 13 subgenera, in 61 municipalities of the state, including the type locality of 9 species, and the first report of Lutzomyia cruzi (Mangabeira 1938), vector of Leishmania infantum (Nicolle 1937), in the Pantanal region of Miranda-Abobral. Other vector species, Bichromomyia flaviscutellata (Mangabeira 1942), Lutzomyia longipalpis (Lutz & Neiva 1912), Nyssomyia antunesi (Coutinho 1939), Nyssomyia intermedia (Lutz & Neiva 1912), Nyssomyia neivai (Pinto 1926), and Nyssomyia whitmani (Antunes & Coutinho 1939), are reported in the entire area of the state. Maps containing the distribution of sandflies on Mato Grosso do Sul biomes were produced. The eclecticism of the species in relation to their ecotypes was observed, with several species using the three biome types as habitats and breeding sites., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Canine obesity is associated with genetic, environmental, and behavioural factors, with the latter including both the behaviour of the dog and the owner. Knowledge about owner perception of canine obesity and its treatment can inform the development of new strategies to help prevent and manage this disease. Therefore, the aim of this study was to evaluate the opinions of dog owners regarding canine obesity and weight management. Dog owners residing in the city of Sao Paulo (Brazil) completed a questionnaire, either at home or in the waiting rooms of 3 veterinary hospitals. Owners determined their dog's body condition score (BCS), and this was compared with BCS determined by a veterinarian. Questionnaire findings from dogs that were in overweight (BCS 6-7/9) or obese (BCS (8-9/9) condition were compared with those in ideal weight (4-5/9) using chi-square tests and odds ratios. A total of 926 dogs were included, of which 480 (52%), 317 (34%) and 129 (14%) were in ideal, overweight and obese condition, respectively. Many owners under-estimated their dog's weight status, with the proportion increasing as the dog's weight status increased (ideal 60/480, 13%; overweight 174/317, 55%; obese 88/129, 68%; P<0.001). Although most owners (890/926, 96%) believed that canine obesity could pose health risks, the proportion that disagreed increased as weight status increased (ideal 12/480, 2%; overweight 14/317, 4%; 10/129, 8%; P = 0.006). Finally, although most owners (880/926, 95%) stated that they would let their dog undergo weight management, only a minority (182/926; 20%) believed that a trained professional was needed, and they had various misperceptions including potential cost and what the strategies that would be effective. Based on the findings of this study, it would be advisable for veterinarians to spend time addressing these misperceptions, in the hope of both improving awareness of obesity and the outcomes of weight management., Competing Interests: The authors have declared that no competing interests exist.

This study aimed to evaluate the accuracy of disk diffusion and Etest methods, compared to that of the broth dilution reference method for identifying beta-lactam susceptibilities of Penicillin-Resistant, Ampicillin-Susceptible Enterococcus faecalis (PRASEF) isolates. Fifty-nine PRASEF and 15 Penicillin-Susceptible, Ampicillin-Susceptible E. faecalis (PSASEF) clinical nonrepetitive isolates were evaluated. The effectiveness of five beta-lactams (ampicillin, amoxicillin, imipenem, penicillin, and piperacillin) was tested. All antimicrobial susceptibility tests were performed and interpreted according to the Clinical and Laboratory Standards Institute guidelines. Interpretative discrepancies, such as essential agreement, categorical agreement, and errors, were assessed. The acceptability was ≥ 90% for both categorical agreement and essential agreement. Etest proved to be an accurate method for testing beta-lactam susceptibilities of the emerging PRASEF isolates, disk diffusion presented poor performance, particularly for imipenem and piperacillin.

Background: Canine obesity is the cause of several health issues, and may predispose other diseases, such as orthopaedic disorders, endocrinopathies, metabolic abnormalities and cardiorespiratory disease which can lead to a decreased quality of life and reduced lifespan. Dog are considered overweight when their body weight is ≥15% above their ideal body weight, and as obese when their body weight exceeds 30% of optimal. Prevalence of canine obesity is estimated to be around 5%-20%, and up to 30%-40% when all overweight dogs are considered. Treatment is based on weight loss programmes, focused on caloric restriction associated with exercise. However, success rate of treatment is low., Objectives: The aim of this study was to investigate the reasons for weight loss failure of obese dogs during treatment with low-calorie diets., Methods: Records of obese dogs undergoing weight loss programmes between May 2014 and May 2017, assessed by a team specialized in veterinary nutrition, were retrospectively evaluated. Dogs were classified according to weekly weight loss rate (WWLR) (<1%, between 1% and 2%, and more than 2%) and owner compliance with a dietary prescription and physical activity recommendations., Results: The mean WWLR was not satisfactory (<1%) in 64.4% (n = 47/73) of dogs. Regarding owner compliance with the dietary prescription, 44.7% (n = 21/47) of owners did not follow diet prescriptions and physical activity recommendations for their dogs. There was a significant association (p = .01) between compliance of owners and satisfactory weight loss rate; however, there was no association between weight loss success, diet composition sex, reproductive status, age of the dogs and their physical activity (p ≥ .05)., Conclusions: Non-compliance represents a great challenge in the treatment of canine obesity, and may be of more importance than other aspects., (© 2019 The Authors. Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Introduction: Hippotherapy allows the development of affective, sensory, motor, and cognitive areas, besides providing the practitioner with several movements and stimuli necessary for therapeutic progress. However, there is a limited amount of scientific evidence regarding the suitability of the mount material, mount type, and hippotherapy session duration, as well as regarding the activation of specific muscle groups during the practice and its applicability to activities of daily living., Objective: This study aimed to study the neuromuscular activation behavior of the iliocostalis, longissimus, multifidus, and upper trapezius muscles of children during four hippotherapy session time points using a functional task. It also compared two different mount materials for riding., Methodology: A total of 30 children were randomly assigned to one of three groups: Saddle Hippotherapy Group, Blanket Hippotherapy Group, and Control Group. Data were collected with an electromyograph in a functional task that comprised trunk movements to pick up an object. Assessments took place at four times during the session., Results: There was a significant increase in the neuromuscular activation of the iliocostalis, longissimus, and multifidus muscles after a 30-min session. The trapezius muscle showed increased neuromuscular activation after only 10 min. It continued to increase (but without a statistical difference) after and 20 and 30 min., Conclusion: Hippotherapy promoted neuromuscular activation of the trunk muscles in children, assessed through a functional task, and was influenced by both session time and mount material. Specifically, the greater neuromuscular performance occurred when an exercise was performed using saddle and stirrup and lasted 30 min., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The use of the pulsed current can be an alternative to decrease the electrode polarization, as well as achieving lower energy consumption. This study investigated the electrocoagulation through pulsed current for the removal of natural organic matter from water. The experiments were carried out using Box-Behnken factorial design with the response surface methodology for the design of experiments, modeling and interpreting of the results. The electrocoagulation cell consisted of an acrylic reactor with 4 L capacity with four electrodes of aluminum, in parallel connection mode. The experimental independent variables studied were: current density (5.5 to 44.5 A m -2 ), electrodes spacing (2 to 7.6 mm), stirring rate (200 to 1,000 rpm), frequency (500 to 5,000 Hz), humic acid concentration (5 to 20 mg L -1 ) and NaCl (100 to 300 mg L -1 ) as supporting electrolyte, evaluating the residual apparent color (RAC) and electric energy consumption (EEC). The pH of the solution increased during the experiments, reaching basic values. The response surface regression procedure was employed to fit the second-order polynomial, and the model fitted well to the obtained values, reaching R 2 0.9995 (RAC) and R 2 0.9989 (EEC). The lowest RAC was 11.8 Hazen units (96.2% color removal), where the EEC was 0.393 kWh m -3 .

Aim: To compare the pathogenesis of vulvovaginal candidiasis by three Candida species in diabetic mice. Materials & methods: Estrogenized and diabetic mice were challenged with C. albicans , C. tropicalis and C. glabrata . Results: Diabetic animals infected with C. albicans and C. tropicalis maintained the highest fungal burden, despite of high levels of proinflammatory cytokines (IL-6 and TNF-α), respectively. For C. glabrata , the results were similar in diabetic and nondiabetic groups. Conclusion: C. tropicalis was as invasive as C. albicans , and both were more effective than C. glabrata . This ability was attributed to filamentation, which may be stimulated by glucose levels from vaginal fluid. In addition, the high burden may be attributed to the apparent immunological inefficiency of the diabetic host.

Automated acoustic recognition of birds is considered an important technology in support of biodiversity monitoring and biodiversity conservation activities. These activities require processing large amounts of soundscape recordings. Typically, recordings are transformed to a number of acoustic features, and a machine learning method is used to build models and recognize the sound events of interest. The main problem is the scalability of data processing, either for developing models or for processing recordings made over long time periods. In those cases, the processing time and resources required might become prohibitive for the average user. To address this problem, we evaluated the applicability of three data reduction methods. These methods were applied to a series of acoustic feature vectors as an additional postprocessing step, which aims to reduce the computational demand during training. The experimental results obtained using Mel-frequency cepstral coefficients (MFCCs) and hidden Markov models (HMMs) support the finding that a reduction in training data by a factor of 10 does not significantly affect the recognition performance., Competing Interests: The authors declare there are no competing interests., (©2020 de Oliveira et al.)

Ecofriendly nanostructured materials have been proposed as promising alternative control means to prevent plant diseases. Chitosan nanoparticles (CN), silica nanoparticles (SN) and chitosan-silica nanocomposites (CSN) were synthesized and their morphology and structure was characterized by transmission electron microscope (TEM), scanning electron microscopy (SEM), infrared spectra (FT-IR) and Raman spectroscopy. Their antifungal efficiency against Botrytis cinerea, the causal fungus of gray mold disease of table grapes, was tested in vitro and in vivo (under artificial and natural infections). In vitro tests showed that CN, SN and CSN reduced fungal growth by 72, 76 and 100%, respectively at 1% as compared to control. Under natural infection, at the end of cold storage, CSN was the most effective treatment, and reduced the development of gray mold by 59 and 83%, for 'Italia' and 'Benitaka' grapes, respectively as compared to the water control. Results indicate that a synergistic effect of CSN against gray mold was observed. The impact of tested nanocomposites on soluble solids - TSS, titratable acidity - TA, TSS/TA, berry color, mass loss, stem browning and shattered berries was investigated. No negative effect of tested nanomaterials in term of grape quality was observed. For 'Italia' table grape, CN and CSN can preserve bunches from mass loss as compared to control. Also, the effect of CSN on reactive species of oxygen (ROS), ATP content and mitochondrial membrane potential (MMP) of B. cinerea spores was determined to verify its mode of action. The obtained results suggested CSN, as alternative control means, to reduce/substitute the use of fungicides to control gray mold of table grapes while maintaining grape quality., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Serodiagnosis of Leishmania infantum infection in dogs relies on the detection of antibodies against leishmanial crude extracts or parasitic defined antigens. The expansion of canine leishmaniasis from geographical areas of Brazil in which the infection is endemic to regions in which the disease is emerging is occurring. This fact makes necessary the analysis of the serodiagnostic capabilities of different leishmanial preparations in distinct geographical locations. In this article sera from dogs infected with Leishmania and showing the clinical form of the disease, were collected in three distinct Brazilian States and were tested against soluble leishmanial antigens or seven parasite individual antigens produced as recombinant proteins. We show that the recognition of soluble leishmanial antigens by sera from these animals was influenced by the geographical location of the infected dogs. Efficacy of the diagnosis based on this crude parasite preparation was higher in newly endemic regions when compared with areas of high disease endemicity. We also show that the use of three of the recombinant proteins, namely parasite surface kinetoplastid membrane protein of 11 kDa (KMP-11), and two members of the P protein family (P2a and P0), can improve the degree of sensitivity without adversely affecting the specificity of the diagnostic assays for canine leishmaniasis, independently of the geographical area of residence. In addition, sera from dogs clinically healthy but infected were also assayed with some of the antigen preparations. We demonstrate that the use of these proteins can help to the serodiagnosis of Leishmania infected animals with subclinical infections. Finally, we propose a diagnostic protocol using a combination of KMP-11, P2a y P0, together with total leishmanial extracts., (© 2019 The Authors.)

Leucine can stimulate protein synthesis in skeletal muscle, and recent studies have shown an increase in leucine-related mitochondrial biogenesis and oxidative phosphorylation capacity in muscle cells. However, leucine-related effects in tumour tissues are still poorly understood. Thus, we described the effects of leucine in both in vivo and in vitro models of a Walker-256 tumour. Tumour-bearing Wistar rats were randomly distributed into a control group (W; normoprotein diet) and leucine group (LW; leucine-rich diet [normoprotein + 3% leucine]). After 20 days of tumour evolution, the animals underwent 18 -fludeoxyglucose positron emission computed tomography ( 18 F-FDG PET-CT) imaging, and after euthanasia, fresh tumour biopsy samples were taken for oxygen consumption rate measurements (Oroboros Oxygraph), electron microscopy analysis and RNA and protein extraction. Our main results from the LW group showed no tumour size change, lower tumour glucose ( 18 F-FDG) uptake, and reduced metastatic sites. Furthermore, leucine stimulated a shift in tumour metabolism from glycolytic towards oxidative phosphorylation, higher mRNA and protein expression of oxidative phosphorylation components, and enhanced mitochondrial density/area even though the leucine-treated tumour had a higher number of apoptotic nuclei with increased oxidative stress. In summary, a leucine-rich diet directed Walker-256 tumour metabolism to a less glycolytic phenotype profile in which these metabolic alterations were associated with a decrease in tumour aggressiveness and reduction in the number of metastatic sites in rats fed a diet supplemented with this branched-chain amino acid.