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Marina F Côrtes,1 Ana MN Botelho,1 Luiz GP Almeida,2 Rangel C Souza,2 Oberdan de Lima Cunha,2 Marisa F Nicolás,2 Ana TR Vasconcelos,2 Agnes MS Figueiredo1 1Laboratory of Molecular Biology of Bateria, Department of Medical Microbiology, Paulo de Goes Institute of Microbiology, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil; 2National Laboratory of Scientific Computing, Bioinformatics Laboratory, Petropolis, Rio de Janeiro, Brazil Abstract: A pivotal event in the evolutionary path of methicillin-resistant Staphylococcus aureus (MRSA) is the acquisition of the staphylococcal cassette chromosome mec (SCCmec) element carrying the mecA gene, the determinant of methicillin resistance. Community-acquired (CA) MRSA is commonly associated with skin/soft tissue infections, and doxycycline is one of the drug choices for this purpose. Doxycycline resistance is associated with the acquisition of the tetK gene carried by the S. aureus plasmid pT181, which may also be integrated into SCCmec III and V. The aim of this study was to describe a novel SCCmec IV subtype (IVm) carrying tetK and reveal the genetic context of this element. The SCCmec sequence was obtained by whole-genome sequencing of the MRSA strain 2288 (ST1 CA-MRSA) and genomic analysis performed using different bioinformatics tools. A copy of pT181 was found to be integrated in the new SCCmec IVm of the strain 2288. The SCCmec IVm has high nucleotide identity (99%) with SCCmec IVa of the strain MW2, except for the J3 region, where the pT181 – carrying tetK gene – is inserted. Inverted repeats (IRs) flanking pT181 were found in this region, suggesting the occurrence of recombination events. The strain 2288 (spa type t125) shares most of the virulence attributes with MW2 (spa type t128), which is recognized in the past as a cause of severe infections in children in USA. The pattern of branching in the phylogenetic tree depicts a recent common ancestor shared by the 2228 strain and other MRSA from USA, including ERS410852, TCH70, CIG1835, CO-41, MW2, and USA400-0051, but none of them carried pT181. This study also showed that the tetK carried by SCCmec IVm is functional, determining resistance to doxycycline and tetracycline. The potential dissemination of the tetK and mecA genes in the same genetic event by the acquisition of this new SCCmec subtype is of concern for community infections. Keywords: MRSA, mec cassette, CA-MRSA, doxycycline resistance

Thioredoxin-1 is a ubiquitous protein involved in phenotypical and functional changes in dendritic cells (DC). We investigated the effect of lipopolysaccharide (LPS), skin sensitizers, and irritants on thioredoxin-1 by Western blot and immunofluorescence and on mRNA by real-time PCR. As DC models, we used a skin DC line and DC derived from human blood monocytes. We observed that all tested chemicals increased thioredoxin-1 expression, which is only transient for irritants, being the strongest effect observed for LPS (63 ± 15%). To address the involvement of thioredoxin-1 in DC maturation, we analysed the effect of an activator of thioredoxin-1 expression, hydrogen peroxide, on CD86 expression, a marker of DC maturation. We found that hydrogen peroxide increases thioredoxin-1 and CD86 expression reinforcing thioredoxin-1 involvement in DC maturation. Because mitogen-activated protein kinases and PI3K are activated upon DC maturation, we also analysed their involvement in thioredoxin-1 modulation. We verified that LPS-induced upregulation of thioredoxin-1 expression was dependent on PI3K pathway. [ABSTRACT FROM AUTHOR]

Presents a clinical study on ectrodactyly-ectodermal dysplasia-clefting syndrome in a boy. Presence of ectrodactyly and hypotrichosis in the father of the boy; Involvement of urogenital anomalies in the syndrome; Ectodermal components of the syndrome.

This article reports on contact and photocontact dermatitis caused due to sunscreens. Sunscreens are widely used to protect the skin from harmful effects of ultraviolet (UV) light. but do not completely prevent photoaging. photocarcinogenesis or photoimmunosuppression. Contact with sunscreens has also caused pruritus or burning facial erythema, contact urticaria and allergic and photoallergic contact dermatitis. It has been observed that many patients show positive reactions to sunscreen lotions.

Objective: To systematically review the effects of recruitment maneuvers on patients with acute respiratory distress syndrome., Methods: This systematic review and meta-analysis using the PICO methodology with keywords (respiratory distress syndrome, recruitment maneuvers, lung recruitment, acute respiratory distress syndrome, alveolar recruitment, and adult acute respiratory distress syndrome). Studies involving patients >18 years, regardless of sex, with acute respiratory distress syndrome, mechanically ventilated for at least 24 h, published in English, Portuguese, and Spanish, with no year restrictions, were included. Studies that combined recruitment maneuvers with other techniques and those conducted in animals were excluded. Boolean operators "AND" and "OR" were used., Results: Fifteen studies were included. The recruitment maneuver proved to be effective in oxygenating patients (mean difference=45.05 mmHg (95% confidence interval (95%CI): 31.37-58.74)), but there was no statistically significant difference in the rate of mortality OR=0.89 (95%CI=0.74-1.08) and barotrauma RR=0.93 (95%CI=0.56-1.54)., Conclusion: Recruitment maneuvers should not be used routinely in the care of patients with acute respiratory distress syndrome, but it is a good rescue strategy when other methods fail to improve oxygenation., Prospero Database Registration: (www.crd.york.ac.uk/prospero) under ID CRD42021227231.

Introduction: Intestinal microeukaryote parasites are major contributors to the burden of diarrhoea in humans and domestic animals, but their epidemiology in wildlife is not fully understood. We investigated the frequency, genetic diversity and zoonotic potential of protists of animal and public health significance in free-ranging grey wolf (Canis lupus) populations in south-western Europe., Methods: Individually formed faecal samples collected from necropsied wolves or scat trails in Italy (n = 47), Portugal (n = 43) and Spain (n = 225) during the period 2011-2023 were retrospectively analysed using molecular (PCR and Sanger sequencing) methods. Complementary epidemiological data were gathered when available., Results: Giardia duodenalis was the most frequent microeukaryote found (40.3%, 127/315; 95% CI: 34.9-46.0), followed by Cryptosporidium spp. (3.5%, 11/315; 95% CI: 1.8-6.2), Enterocytozoon bieneusi and Encephalitozoon spp. (1.6%, 5/315; 95% CI: 0.5-3.7 each). Blastocystis was not identified in any of the faecal samples analysed. Sequence analyses confirmed the presence of canine-adapted assemblage D within G. duodenalis (n = 7). Three Cryptosporidium species were identified, namely canine-adapted C. canis (n = 9), zoonotic C. parvum (n = 1) and primarily anthroponotic C.hominis (n = 1). Genotyping tools enabled the identification of subtype family XXe2 within C. canis. Among microsporidia, the canine-adapted genotype PtEb IX was identified within E. bieneusi. Two samples were confirmed as Enc. intestinalis and three more as Enc. cuniculi genotype IV. This is the first record of Enc. intestinalis and Enc. cuniculi in the grey wolf globally., Conclusions: Silent carriage of intestinal microeukaryotes seems common in free-ranging grey wolves in southwestern Europe. Wolves can contribute to environmental contamination through the transmission stages (cysts, oocysts, spores) of species/genotypes potentially infective to humans. Individuals in close contact with wolf carcasses or their faecal material may be at potential risk of infection by microeukaryotic pathogens., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Aims: Arteriovenous malformations (AVMs), a disorder characterized by direct shunts between arteries and veins, are associated with genetic mutations. However, the mechanisms leading to AV shunt formation and how shunts can be reverted are poorly understood., Methods and Results: Here, we report that oxygen-induced retinopathy (OIR) protocol leads to the consistent and stereotypical formation of AV shunts in non-genetically altered mice. OIR-induced AV shunts show all the canonical markers of AVMs. Genetic and pharmacological interventions demonstrated that changes in the volume of venous endothelial cells (EC)-hypertrophic venous cells-are the initiating step promoting AV shunt formation, whilst EC proliferation or migration played minor roles. Inhibition of the mTOR pathway prevents pathological increases in EC volume and significantly reduces the formation of AV shunts. Importantly, we demonstrate that ALK1 signalling cell-autonomously regulates EC volume in pro-angiogenic conditions, establishing a link with hereditary haemorrhagic telangiectasia-related AVMs. Finally, we demonstrate that a combination of EC volume control and EC migration is associated with the regression of AV shunts., Conclusion: Our findings highlight that an increase in the EC volume is the key mechanism driving the initial stages of AV shunt formation, leading to asymmetric capillary diameters. Based on our results, we propose a coherent and unifying timeline leading to the fast conversion of a capillary vessel into an AV shunt. Our data advocate for further investigation into the mechanisms regulating EC volume in health and disease as a way to identify therapeutic approaches to prevent and revert AVMs., Competing Interests: Conflict of interest: none declared., (© The Author(s) 2024. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Cellulose nanocrystals (CNCs) are known for several features which make them one of the nanoparticles with great potential for application in different areas such as biosensors development and carrier molecules. However, there are few reports in the literature describing the coupling analysis of cellulose nanocrystals with biomolecules, and the existing ones are laborious, still needing a large laboratory structure. In this paper, we describe the non-covalent coupling of Bovine Serum Albumin protein (BSA) to CNCs using Green Fluorescent Protein (GFP) as a reporter molecule. Analyzes of these tests were performed using the flow cytometry technique. The nanocrystal particles became fully fluorescent when coupled to a concentration of 1 µg GFP. BSA coupling was able to block binding to GFP when at a concentration of 8 µg. Thus, flow cytometry has proven to be a simple and effective methodology for determining BSA coupling percentage to CNCs.

Recent molecular and metagenomic studies have revealed that the obligate anaerobic protist Blastocystis is found more prevalently and with higher subtype diversities in herbivore species than in carnivore species. However, information on wild carnivore species is scarce. Here, we investigated the presence of Blastocystis by molecular methods in fecal DNA samples of free-ranging and captive Iberian lynxes from Spain (n = 243) and Portugal (n = 30). In addition, a systematic review was conducted to obtain information on the Blastocystis prevalence rates and subtype diversities reported in free-living and captive wild carnivores worldwide during the period 2000-2024. Blastocystis was not detected by PCR in any of the samples investigated. Analyses of the data gathered from our systematic review revealed that Blastocystis is uncommon either in free-living (2.1 %, 29/1377) or captive (8.5 %, 100/1175) wild carnivore species. Many of these findings seem to result from accidental acquisition via prey animals, scavenging, contaminated water/feed (free-ranging wild carnivores), or cross-species transmission among animals sharing enclosures (captive wild carnivores). Comparative metagenomic studies analyzing gut microbiota profiles of carnivores are needed to fully understand how microbial communities affect Blastocystis colonization., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2025 Elsevier Ltd. All rights reserved.)

Intrinsically disordered proteins (IDPs) often contain proline residues that undergo cis/trans isomerization. While molecular dynamics (MD) simulations have the potential to fully characterize the proline cis and trans subensembles, they are limited by the slow timescales of isomerization and force field inaccuracies. NMR spectroscopy can report on ensemble-averaged observables for both the cis-proline and trans-proline states, but a full atomistic characterization of these conformers is challenging. Given the importance of proline cis/trans isomerization for influencing the conformational sampling of disordered proteins, we employed a combination of all-atom MD simulations with enhanced sampling (metadynamics), NMR, and small-angle x-ray scattering (SAXS) to characterize the two subensembles of the ORF6 C-terminal region (ORF6 CTR ) from SARS-CoV-2 corresponding to the proline-57 (P57) cis and trans states. We performed MD simulations in three distinct force fields: AMBER03ws, AMBER99SB-disp, and CHARMM36m, which are all optimized for disordered proteins. Each simulation was run for an accumulated time of 180-220 μs until convergence was reached, as assessed by blocking analysis. A good agreement between the cis-P57 populations predicted from metadynamic simulations in AMBER03ws was observed with populations obtained from experimental NMR data. Moreover, we observed good agreement between the radius of gyration predicted from the metadynamic simulations in AMBER03ws and that measured using SAXS. Our findings suggest that both the cis-P57 and trans-P57 conformations of ORF6 CTR are extremely dynamic and that interdisciplinary approaches combining both multiscale computations and experiments offer avenues to explore highly dynamic states that cannot be reliably characterized by either approach in isolation., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

The ongoing increase in wild boar populations across Europe has fostered human-wildlife conflicts, including the transmission of emerging pathogens with zoonotic importance. Blastocystis is a ubiquitous, faecal-oral transmitted protist that can cause gastrointestinal illnesses and is observed in humans and animals worldwide. The role of wildlife in the epidemiology of Blastocystis is insufficiently understood. Thus, we investigated the occurrence and subtype diversity of Blastocystis in free-ranging wild boars from the Iberian Peninsula using conventional PCR and next-generation amplicon sequencing of a fragment of the ssu RNA gene. A total of 459 wild boar faecal samples were collected across Spain (n = 360) and Portugal (n = 99) between 2014 and 2021. Blastocystis was present in 15.3% (70/459; 95% CI 12.1-18.9) of the wild boars analysed, and its occurrence was significantly higher in Portugal (34.3%, 34/99; 95% CI 25.1-44.6) than in Spain (10.0%, 36/360; 95% CI 7.1-13.6). Seven Blastocystis subtypes (ST5, ST10b, ST13-ST15, ST24b, and ST43) were detected among the surveyed wild boar populations, with greater variability detected in Portuguese samples. ST5 was identified in all the Blastocystis-positive animals, whereas 14.3% of them harboured ST mixed colonisations. Our results demonstrate that Blastocystis ST5 is particularly adapted to infect wild boars. The additional identification of zoonotic STs reinforces the role of wild boars as spreaders of zoonotic infections with public health significance., (© 2024. The Author(s).)

Cyclisation of peptides by forming thioether (lanthionine), disulfide (cystine) or methylene thioacetal bridges between side chains is established as an important tool to stabilise a given structure, enhance metabolic stability and optimise both potency and selectivity. However, a systematic comparative study of the effects of differing bridging modalities on peptide conformation has not previously been carried out. In this paper, we have used the NMR deconvolution algorithm, NAMFIS, to determine the conformational ensembles, in aqueous solution, of three cyclic analogues of angiotensin(1-7), incorporating either disulfide, or non-reducible thioether or methylene thioacetal bridges. We demonstrate that the major solution conformations are conserved between the different bridged peptides, but the distribution of conformations differs appreciably. This suggests that subtle differences in ring size and bridging structure can be exploited to fine-tune the conformational properties of cyclic peptides, which may modulate their bioactivities., (© 2024 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)

Most proteins fold during biosynthesis on the ribosome 1 , and co-translational folding energetics, pathways and outcomes of many proteins have been found to differ considerably from those in refolding studies 2-10 . The origin of this folding modulation by the ribosome has remained unknown. Here we have determined atomistic structures of the unfolded state of a model protein on and off the ribosome, which reveal that the ribosome structurally expands the unfolded nascent chain and increases its solvation, resulting in its entropic destabilization relative to the peptide chain in isolation. Quantitative 19 F NMR experiments confirm that this destabilization reduces the entropic penalty of folding by up to 30 kcal mol -1 and promotes formation of partially folded intermediates on the ribosome, an observation that extends to other protein domains and is obligate for some proteins to acquire their active conformation. The thermodynamic effects also contribute to the ribosome protecting the nascent chain from mutation-induced unfolding, which suggests a crucial role of the ribosome in supporting protein evolution. By correlating nascent chain structure and dynamics to their folding energetics and post-translational outcomes, our findings establish the physical basis of the distinct thermodynamics of co-translational protein folding., (© 2024. The Author(s).)

Peruvian fava beans (PFB) are used in traditional cuisine as a nutrient-rich, flavorful, and textural ingredient; however, little is known about their industrial properties. This study evaluated the physicochemical, nutritional, and techno-functional characteristics of PFB varieties: Verde, Quelcao, and Peruanita. PFB exhibited distinct physical characteristics, quality parameters, and morphology. The color patterns of the seed coat and the hardness were the main parameters for distinguishing them. Nutritionally, all three samples exhibited high protein (23.88-24.88 g/100 g), with high proportion of essential amino acids, high dietary fiber (21.74-25.28 g/100 g), and mineral content. They also contain polyphenols (0.79-1.25 mg GAE/g) and flavonoids (0.91-1.06 mg CE/g) with antioxidant potential (16.60-21.01 and 4.68-5.17 µmol TE/g for ABTS and DPPH assays, respectively). Through XRD measurements, the semi-crystalline nature of samples was identified, belonging to the C-type crystalline form. Regarding techno-functionality, PFB flours displayed great foaming capacity, with Verde variety being the most stable. Emulsifying capacity was similar among samples, although Peruanita was more stable during heating. Upon heating with water, PFB flours reached peak viscosities between 175 and 272 cP, and final viscosities between 242 and 384 cP. Quelcao and Verde formed firmer gels after refrigeration. Based on these results, PFB would be useful to developing innovative, nutritious, and healthy products that meet market needs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Blastocystis is a ubiquitous intestinal protist in humans and animals worldwide. The traditional livestock free-roaming raising system in rural communities increases the risk of infection with contact with a wider range of pathogens transmitted via the faecal-oral route associated with that wildlife-livestock-human interface. However, no studies have been conducted to determine the occurrence and subtype distribution of Blastocystis in livestock in Portugal. Here, we collected 180 faecal samples from herbivore livestock (cattle, goats, horses, and sheep) in different regions of the country to investigate Blastocystis prevalence and subtype diversity using PCR and next-generation amplicon sequencing. Blastocystis was present in 40.6% (73/180; 95% CI: 33.31-48.11) of the samples (goats, 81.0%; sheep, 60.9%; cattle, 32.2%). None of the horse samples were Blastocystis-positive. Eighteen subtypes were detected (ST1-ST3, ST5-ST7, ST10, ST13, ST14, ST21, ST23-ST26, ST30, ST42-ST44). Mixed infections were detected in 97.3% of the Blastocystis-positive samples. Potentially zoonotic subtypes were identified in 75.0%, 96.4%, and 100% of the Blastocystis-positive specimens collected from cattle, sheep, and goats, respectively. These results demonstrate that cattle, sheep, and goats harbour a high diversity of Blastocystis subtypes in the study regions. Importantly, our data provide novel molecular evidence strongly suggesting that some Blastocystis STs/ST subgroups may have differential host specificity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: To evaluate the effectiveness of vaccines developed against covid-19 in reducing mortality in people hospitalized with severe acute respiratory syndrome (SARS) caused by SARS-CoV-2., Methods: This is a retrospective cohort that evaluated risk factors and the effectiveness of the two-dose vaccination schedule in reducing the mortality of people hospitalized for covid-19 in the state of Paraíba from February to November 2021. The explanatory variables were vaccination status, presence of comorbidities, socioeconomic and demographic characteristics. Descriptive analyses and bivariate and multivariable logistic regression were performed., Results: Most hospitalizations and deaths occurred until May 2021. The percentage of patients with a complete vaccination schedule was similar across patients admitted to public and private hospitals and higher in residents of less developed municipalities. Multivariable analysis demonstrated that women (OR = 0.896; 95%CI 0.830-0.967) and people admitted to private hospitals (OR = 0.756; 95%CI 0.679-0.842) were less likely to die. Presence of any comorbidity (OR = 1.627; 95%CI 1.500-1.765) and age ≥ 80 years (OR = 7.426; 95%CI 6.309-8.741) were risk factors for death. Patients with complete vaccination schedule at the time of admission were 41.7% less likely to die (OR = 0.583; 95% CI 0.501-0.679) from covid-19 in the adjusted analysis, as compared to unvaccinated patients., Conclusions: The study reveals that immunization was effective in reducing the likelihood of death from covid-19. The results suggest that greater vaccination coverage in the first half of 2021 would prevent thousands of deaths in the country.

Intrinsically disordered proteins (IDPs) are highly dynamic biomolecules that rapidly interconvert among many structural conformations. These dynamic biomolecules are involved in cancers, neurodegeneration, cardiovascular illnesses, and viral infections. Despite their enormous therapeutic potential, IDPs have generally been considered undruggable because of their lack of classical long-lived binding pockets for small molecules. Currently, only a few instances are known where small molecules have been observed to interact with IDPs, and this situation is further exacerbated by the limited sensitivity of experimental techniques to detect such binding events. Here, using experimental nuclear magnetic resonance (NMR) spectroscopy 19 F transverse spin-relaxation measurements, we discovered that a small molecule, 5-fluoroindole, interacts with the disordered domains of non-structural protein 5A from hepatitis C virus with a K d of 260 ± 110 μM. Our analysis also allowed us to determine the rotational correlation times (τ c ) for the free and bound states of 5-fluoroindole. In the free state, we observed a rotational correlation time of 27.0 ± 1.3 ps, whereas in the bound state, τ c only increased to 46 ± 10 ps. Our findings imply that it is possible for small molecules to engage with IDPs in exceptionally dynamic ways, in sharp contrast to the rigid binding modes typically exhibited when small molecules bind to well-defined binding pockets within structured proteins.

The environmental impacts caused by fish farming can lead to changes in aquatic ecosystems, especially in fish communities. In this study, we investigated possible changes in water quality, in the number of species and individuals of the same species caused by the construction of fish nurseries in dammed Amazonian streams. For this purpose, four streams located in the municipalities of Novo Airão and Presidente Figueiredo were selected. Samples were collected in streams without fish farming and in streams with stretches used for fish farming, where collections took place both downstream and upstream. The fish were captured, identified, quantified and the water was collected for physical and chemical analyses, in addition to the environmental characterization that was carried out. Comparisons were made using fish richness and abundance indices between the sampled points and correspondence analysis of the species identified at the collection sites, using the Mantel test and principal component analysis (PCA) for the environmental characteristics of the water and variables physical and chemical, and permutational multivariate analysis of variance (PERMANOVA) to verify the possible effects of species composition on the sampled conditions and on the analyzed environmental variables. In total, 2,302 fish belonging to 53 species, 15 families and six orders were found. We conclude that the Amazonian streams dammed for fish farming, under the analyzed conditions, can be characterized as environments with different and specific levels of richness and abundance, however, the data set analyzed in this study did not show that these characteristics are directly related to productive activity. We recommend that further studies be carried out following the current research.

m6A methylation provides an essential layer of regulation in organismal development, and is aberrant in a range of cancers and neuro-pathologies. The information encoded by m6A methylation is integrated into existing RNA regulatory networks by RNA binding proteins that recognise methylated sites, the m6A readers. m6A readers include a well-characterised class of dedicated proteins, the YTH proteins, as well as a broader group of multi-functional regulators where recognition of m6A is only partially understood. Molecular insight in this recognition is essential to build a mechanistic understanding of global m6A regulation. In this study, we show that the reader IMP1 recognises the m6A using a dedicated hydrophobic platform that assembles on the methyl moiety, creating a stable high-affinity interaction. This recognition is conserved across evolution and independent from the underlying sequence context but is layered upon the strong sequence specificity of IMP1 for GGAC RNA. This leads us to propose a concept for m6A regulation where methylation plays a context-dependent role in the recognition of selected IMP1 targets that is dependent on the cellular concentration of available IMP1, differing from that observed for the YTH proteins., (© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Introduction: Various initiatives to improve access to health care have been implemented internationally. In Brazil, this has included policies intended to expand access to higher education for underrepresented socio-economic groups. These measures have reduced inequalities in access to medical education, but it is not known whether they influence career choices. We examine the effect of these educational policies on physician practice patterns in primary care and/or in medically underserved areas., Methods: This is a cross-sectional study that analysed the association between affirmative educational policies and the career choices of physicians who graduated between 2010 and 2015. Multivariate binary regression analysis was used to evaluate the impact of affirmative educational policies on physician career choices, including primary care practice, practice in cities with ≤20 000 inhabitants, and practice in less developed municipalities 4 years after graduation., Results: We identified the practice patterns of 65 304 (82.8%) physicians 4 years after their graduation. Most physicians included in our analysis were female (54.5%), aged ≤27 years (72.4%), born in developed cities and studied in private medical schools. Physicians admitted to medical school based on racial or social access policies were more likely to practice in municipalities with fewer than 20 000 inhabitants (odds ratio [OR]: 1.64; 1.36-1.98) and in primary care (OR: 1.55; 1.35-1.53). Physicians who benefited from scholarships for under-represented socio-economic groups were more likely to practice in small cities (OR: 1.24; 1.07-1.43) and primary care (OR: 1.23; 1.11-1.37). The provision of financial aid also improved the likelihood of practice in primary care and underserved areas. Graduation from medical schools located in smaller cities was associated with practice in municipalities ≤20 000 inhabitants and primary care., Conclusions: Our results demonstrated that educational policies implemented in the Brazilian context are effective in reducing inequities in physician distribution and led to an increase in the number of physicians practicing in primary care., (© 2022 Association for the Study of Medical Education and John Wiley & Sons Ltd.)

The shortage of physicians in rural and underserved areas is an obstacle to the implementation of Universal Health Coverage (UHC). We carried out a systematic review to analyze the effectiveness of initiatives in medical education aimed to increase the supply of physicians in rural or underserved areas. We searched for studies published between 1999 and 2019 in six databases, following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Interventional or observational controlled studies were defined as inclusion criteria. A total of 955 relevant unique records were selected for inclusion, which resulted in the identification of 17 articles for analysis. The admission of students from rural areas associated with a rural curriculum represented 52.95% of the interventions. Medical practice after graduation in rural or underserved areas was the most evaluated outcome, representing 12 publications (70.59%). Participants of these educational initiatives were more likely to work in rural or underserved areas or to choose family medicine, with significant differences between the groups in 82.35% of the studies. Educational strategies in undergraduate and medical residencies are effective. However, it is necessary to expand these interventions to ensure the supply of physicians in rural or urban underserved areas.

The phylum Microsporidia encompasses a diverse group of obligate, intracellular, and spore-forming organisms able to infect a wide range of animal hosts. Among them, Enterocytozoon bieneusi is the most frequently reported species in humans and animals. Little is known about the presence and epidemiology of E. bieneusi in wildlife. We investigated E. bieneusi occurrence and genetic diversity in wild and domestic mammals, through molecular-detection methods, from different regions across Portugal. A total of 756 samples were collected from 288, 242, and 226 wild carnivores, wild ungulates, and domestic animals, respectively. Overall, eight specimens were E. bieneusi-positive (1.1%, 8/756) obtained from five wild (Iberian lynx, Iberian wolf, red fox, stone marten, and wild boar) and one domestic (sheep) host. Nucleotide sequence analysis identified four genotypes of E. bieneusi, Type IV, Wildboar3, BEB6, and PtEbIX. Three of those genotypes belong to Groups 1 (Type IV and Wildboar3) and 2 (BEB6), which are known to contain genotypes capable of infecting a variety of hosts, including humans, highlighting their public health importance. PtEbIX belongs to the dog-specific Group 11. This study represents the first, largest, and most comprehensive molecular-based epidemiology survey carried out in Portugal in wild and domestic animals to date and the first worldwide identification of E. bieneusi in wolf species. Our study showed that wild carnivores and ungulates may act as reservoirs of zoonotic genotypes of E. bieneusi, establishing their role in maintaining the sylvatic cycle of this parasite while representing a potential source of infection for humans and domestic animals., (© The Author(s) 2023. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

At the end of the nineteenth century, massive population declines were observed in carnivores due to the emergence of infectious diseases. This study aims to investigate, by means of coprological analysis, the prevalence and intensity of the parasites that infect the endangered Iberian wolf Canis lupus signatus and two mesocarnivores (the red fox Vulpes vulpes and the stone marten Martes foina) in Central Portugal. In total, 67.2% of the samples screened were infected; Toxascaris leonina (40.6%) was the parasite with the highest prevalence, followed by Ancylostomatidae and Eimeria spp. (28.1%). Eimeria spp. was found in stone marten with the highest infection rate (37,800 OPG), followed by T. leonina (10,100 EPG) in a red fox sample. Moderate to high levels of parasitic infections were identified in 73.3% of red foxes from the western area. Our results highlight the possibility of cross-infection among these carnivore species and cross-contamination in the wildlife-livestock-human interface., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Background: Typing of staphylococcal cassette chromosome mec (SCCmec) elements is commonly used for studies on the molecular epidemiology of MRSA., Objectives: To perform an investigation centred on uncovering the reasons for misclassification of MRSA clonal complex 5 (CC5) SCCmec type II clinical isolates in our laboratory., Methods: MRSA isolates from CC5 were subjected to WGS and SCCmec typing., Results: This investigation led to the discovery that the classification failure was due to an insertion of IS1272 carrying the fabI gene on a transposable element (TnSha1) that confers increased MIC to the biocide triclosan. Genomic analysis revealed that fabI was present in 25% of the CC5 MRSA isolates sampled. The frequency of TnSha1 in our collection was much higher than that observed among publicly available genomes (0.8%; n = 24/3142 CC5 genomes). Phylogenetic analyses revealed that genomes in different CC5 clades carry TnSha1 inserted in different integration sites, suggesting that this transposon has entered CC5 MRSA genomes on multiple occasions. In at least two genotypes, ST5-SCCmecII-t539 and ST5-SCCmecII-t2666, TnSha1 seems to have entered prior to their divergence., Conclusions: Our work highlights an important misclassification problem of SCCmecII in isolates harbouring TnSha1 when Boye's method is used for typing, which could have important implications for molecular epidemiology of MRSA. The importance of increased-MIC phenotype is still a matter of controversy that deserves more study given the widespread use of triclosan in many countries. Our results suggest expanding prevalence that may indicate strong selection for this phenotype., (© The Author(s) 2022. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

Alpha-1 antitrypsin deficiency, although one of the most common genetic diseases, is a very rare and often undiagnosed cause of panniculitis. The authors present a case characterized by an acute involvement of several areas in the thorax, abdomen, and limbs, occurring after repetitive trauma of the perineal area caused by a long period of cycling. After performing the differential diagnosis and establishing etiology, the patient was started on augmentation therapy with plasma-derived synthetic human alpha-1 proteinase inhibitor and the disease has been under control since then. We recommend lifelong treatment with this medication. At the end of a 10-year follow-up, there has been no evidence of pulmonary emphysema or liver disease. The authors perform a concise review of the genetic and pathogenic mechanisms behind this disease, with a special focus on panniculitis and its treatment., Competing Interests: The authors have no conflicts of interest to declare., (Copyright © 2022 by The Author(s). Published by S. Karger AG, Basel.)

Microsphere-based flow cytometry is a highly sensitive emerging technology for specific detection and clinical analysis of antigens, antibodies, and nucleic acids of interest. In this review, studies that focused on the application of flow cytometry as a viable alternative for the investigation of infectious diseases were analyzed. Many of the studies involve research aimed at epidemiological surveillance, vaccine candidates and early diagnosis, non-infectious diseases, specifically cancer, and emphasize the simultaneous detection of biomarkers for early diagnosis, with accurate results in a non-invasive approach. The possibility of carrying out multiplexed assays affords this technique high versatility and performance, which is evidenced in a series of clinical studies that have verified the ability to detect several molecules in low concentrations and with minimal sample volume. As such, we demonstrate that microsphere-based flow cytometry presents itself as a promising technique that can be adopted as a fundamental element in the development of new diagnostic methods for a number of diseases.

This study assesses the effectiveness of policies to expand access to higher education in reducing inequities in the profile of graduates from medical courses in Brazil. This work consists of a cross-sectional study using data from the 2018 Higher Education Census. Analyses were carried out to identify the association between being a beneficiary of the Quota Law, the University for All Program, or the Student Financing Fund and three markers: born in a small-sized municipality, being non-white, or having attended high school in a public school. Analyses were performed using Pearson's chi-square test, and multivariate analysis was conducted using Poisson regression. A significant association was found between being a beneficiary of these policies and being a non-white graduate, who was born in a small town and who had studied in a public high school. All policies presented adjusted Prevalence Ratio (PR), which demonstrated an association with statistical significance. The Quota Law was the most effective policy, with RP=1.92 for non-white graduates, RP=6.66 for having studied in a public high school, and RP=1.08 for being born in a small town. Despite these results, these groups remain underrepresented in medical courses and in the workforce.

Objectives: to analyze trends in suicide rates in Brazil in the period before and after the start of the economic recession., Methods: interrupted time series research using national suicide data recorded in the period between 2012 and 2017 with socioeconomic subgroups analyses. Quasi-Poisson regression model was employed to analyze trends in seasonally adjusted data., Results: there was an abrupt increase in the risk of suicide after economic recession in the population with less education (12.5%; RR = 1.125; 95%CI: 1.027; 1.232) and in the South Region (17.7%; 1.044; 1.328). After an abrupt reduction, there was a progressive increase in risk for the black and brown population and for those with higher education. In most other population strata, there was a progressive increase in the risk of suicide., Conclusions: the Brazilian economic recession caused different effects on suicide rates, considering social strata, which requires health strategies and policies that are sensitive to the most vulnerable populations.

Myocardial infarction has a high mortality rate worldwide. Therefore, clinical intervention in cardiac remodeling after myocardial infarction is essential. Açai pulp is a natural product and has been considered a functional food because of its antioxidant/anti-inflammatory properties. The aim of the present study was to analyze the effect of açai pulp supplementation on cardiac remodeling after myocardial infarction in rats. After 7 days of surgery, male Wistar rats were assigned to six groups: sham animals fed standard chow (SA0, n = 14), fed standard chow with 2% açai pulp (SA2, n = 12) and fed standard chow with 5% açai pulp (SA5, n = 14), infarcted animals fed standard chow (IA0, n = 12), fed standard chow with 2% açai pulp (IA2, n = 12), and fed standard chow with 5% açai pulp (IA5, n = 12). After 3 months of supplementation, echocardiography and euthanasia were performed. Açai pulp supplementation, after myocardial infarction, improved energy metabolism, attenuated oxidative stress (lower concentration of malondialdehyde, P = 0.023; dose-dependent effect), modulated the inflammatory process (lower concentration of interleukin-10, P<0.001; dose-dependent effect) and decreased the deposit of collagen (lower percentage of interstitial collagen fraction, P<0.001; dose-dependent effect). In conclusion, açai pulp supplementation attenuated cardiac remodeling after myocardial infarction in rats. Also, different doses of açai pulp supplementation have dose-dependent effects on cardiac remodeling., Competing Interests: The authors have declared that no competing interests exist.

In the present work, we examined the morphology, dimensions, cytochemical staining reactions and ultrastructure of blood cells from three freshwater stingray species, Potamotrygon wallacei, Potamotrygon motoro and Paratrygon aiereba, living in the waters of the middle Rio Negro basin (Barcelos, Amazonas, Brazil). We identified erythrocytes, erythroblasts, thrombocytes and four types of leukocytes (basophils, heterophils, lymphocytes and monocytes) in the blood of these stingray species. In all the freshwater stingray species studied, the shapes and dimensions of these cells were similar to those of marine elasmobranchs. Positive PAS staining occurred in heterophils and thrombocytes, and weak staining occurred in lymphocytes and monocytes, while metachromasia only occurred in basophils. Positive Sudan Black B staining was observed in thrombocytes and lymphocytes, and weak staining occurred in heterophils. Basophils and heterophils were the only cells with positive bromophenol blue staining, while no peroxidase staining was observed in any of the four leukocyte types. This is the first study to establish the dimensions and cytochemical staining profiles of blood cells in Amazonian stingray species. Because these elasmobranch species are exported as ornamental fish to countries worldwide, this study can contribute to establishing standards for blood constituents that may be helpful in assessing the health and welfare of these fish in artificial systems., (© 2021. The Author(s).)

Background: The capture of ornamental fish is one of the main economic activities of riverine families in the Amazon. However, studies regarding the local ecological knowledge of workers in this activity are still incipient. In view of this, we have studied and explored the local ecological knowledge of artisanal fishers who specialize in the capture of fish for the aquarium trade in the middle part of the Negro River basin and investigated issues related to the ecological aspects of the fish species that are targeted by this trade in the region., Methods: Therefore, we conducted semi-structured interviews and applied questionnaires to artisanal fishers of ornamental fish (N = 89), from the municipality of Barcelos, from January to April 2016., Results: In total, 41 popular names were cited, which correspond to four ethnocategories and 10 families. The main species were Paracheirodon axelrodi (12.5%), Hemigrammus bleheri (8.3%), Ancistrus dolichopterus (6.4%), Symphysodon discus (5.3%), and Potamotrygon motoro (3.8%). According to the fishers, the species of fish known in the region as "piabas" have a preference for living in clusters (28.9%) and carry out migratory movements (26.1%). The diet of local fish species reported by fisheries is diverse, though mainly based on periphyton (42.2%), and the reproductive cycle directly influenced by the period of flooding of rivers in the region (37.6%) CONCLUSION: Our study revealed that the fishers possess information on the ecological aspects of local ornamental fish species, many of which are consistent with scientific literature. The information presented may assist in the decision-making process for the management of local fishery resources and contribute to the resumption of growth and sustainability in the capture of ornamental fish.

Background: Orange juice (OJ) is rich in polyphenols with anti-inflammatory and antioxidant properties. After myocardial infarction (MI), complex changes occur in cardiac structure and function, which is known as cardiac remodeling (CR). Oxidative stress and inflammation can modulate this process. We hypothesized that the consumption of OJ attenuates the CR after MI., Objectives: To evaluate the influence of OJ on CR after MI by analysis of functional, morphological, oxidative stress, inflammation, and energy metabolism variables., Methods: A total of 242 male rats weighing 200-250 g were submitted to a surgical procedure (coronary artery ligation or simulated surgery). Seven days after surgery, survivors were assigned to one of the four groups 1) SM, sham animals with water and maltodextrin (n= 20); 2) SOJ, sham animals with OJ (n= 20); 3) IM, infarcted animals with water and maltodextrin (n= 40); and 4) IOJ, infarcted animals with OJ (n = 40). Statistical analysis was performed by the two-way ANOVA supplemented by Holm-Sidak. Results are presented as mean ± standard deviation, the level of significance adopted was 5%., Results: After 3 months, MI led to left ventricular (LV) hypertrophy, with systolic and diastolic dysfunction, and increased oxidative stress and inflammatory mediators. OJ intake reduced LV cavity and improved systolic and diastolic function. The OJ animals presented lower activity of glutathione peroxidase and higher expression of heme-oxygenase-1 (HO-1)., Conclusion: OJ attenuated CR in infarcted rats and HO-1 may be play an important role in this process.

The present study objectified to evaluate the trophic relationships and the possible aggressive mimicry involving Carnegiella strigata, Carnegiella marthae and Gnathocharax steindachneri, in lowland forest streams. Samplings occurred in November 2002, March and August 2003 in Amanã Sustainable Development Reserve (ASDR). The total of 943 fish was caught, being 79.43% of C. marthae, 3.18% of C. strigata and 17.39% of G. steindachneri. Stomach contents, degree of stomach repletion, relative volume and frequency of occurrence of food items were evaluated. Thirteen food items were identified, and the presence of terrestrial and aquatic insects indicates the high dependence of these species and the flooded forest. Low feeding overlap was observed between Carnegiella strigata and C. marthae compared to Gnathocharax steindachneri. However, the overlap between the two Carnegiella species was relatively high, which could explain the low frequency of syntopic occurrence among these species, suggesting a possible case of competitive exclusion.

Sprouting angiogenesis is fundamental for development and contributes to cancer, diabetic retinopathy, and cardiovascular diseases. Sprouting angiogenesis depends on the invasive properties of endothelial tip cells. However, there is very limited knowledge on how tip cells invade into tissues. Here, we show that endothelial tip cells use dactylopodia as the main cellular protrusion for invasion into nonvascular extracellular matrix. We show that dactylopodia and filopodia protrusions are balanced by myosin IIA (NMIIA) and actin-related protein 2/3 (Arp2/3) activity. Endothelial cell-autonomous ablation of NMIIA promotes excessive dactylopodia formation in detriment of filopodia. Conversely, endothelial cell-autonomous ablation of Arp2/3 prevents dactylopodia development and leads to excessive filopodia formation. We further show that NMIIA inhibits Rac1-dependent activation of Arp2/3 by regulating the maturation state of focal adhesions. Our discoveries establish a comprehensive model of how endothelial tip cells regulate its protrusive activity and will pave the way toward strategies to block invasive tip cells during sprouting angiogenesis., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Paracoccidioidomycosis (PCM) is a systemic fungal disease caused by Paracoccidioides spp., whose clinical outcome depends on immune response. Interleukin 32 (IL-32) is a cytokine present in inflammatory and infectious diseases, including bacterial, virus and protozoan infections. Its role in fungal disease remains unclear. The axis IL-15, IL-32 and vitamin D leads to microbicidal capacity against intracellular pathogens. Thus, the aims of this study were to investigate the production of IL-32 during Paracoccidioides spp. infection and whether this cytokine and IL-15 can increase P. brasiliensis control in a vitamin D dependent manner. IL-32 was highly detected in oral lesions from patients with PCM. In addition, high production of this cytokine was intracellularly detected in peripheral blood mononuclear cells (PBMCs) from healthy donors after exposure to particulated P. brasiliensis antigens (PbAg). The IL-32γ isoform was predominantly expressed, but there was mRNA alternative splicing for IL-32α isoform. The induction of IL-32 was dependent on Dectin-1 receptor. Infection of PBMCs with P. brasiliensis yeasts did not significantly induce IL-32 production even after activation with exogenous IFN-γ or IL-15 treatments. Although IL-15 was a potent inducer of IL-32 production, treatment with this cytokine did not increase the fungal control unless vitamin D was present in high levels. In this case, both IL-15 and IL-32 increased fungicidal activity of PBMCs. Together, data showed that IL-32 is present in lesions of PCM, PbAg induces IL-32, and the axis of IL-15/IL-32/vitamin D can contribute to control fungal infection. The data suggest that exposure to molecules from P. brasiliensis, as β-glucans, is needed to induce IL-32 production since only heat-killed and sonicated P. brasiliensis yeasts were able to increase IL-32, which was blocked by anti-Dectin-1 antibodies. This is the first description about IL-15/IL-32/vitamin D pathway role in P. brasiliensis infection., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The Program for Biodiversity Research (PPBio) is an innovative program designed to integrate all biodiversity research stakeholders. Operating since 2004, it has installed long-term ecological research sites throughout Brazil and its logic has been applied in some other southern-hemisphere countries. The program supports all aspects of research necessary to understand biodiversity and the processes that affect it. There are presently 161 sampling sites (see some of them at Supplementary Appendix), most of which use a standardized methodology that allows comparisons across biomes and through time. To date, there are about 1200 publications associated with PPBio that cover topics ranging from natural history to genetics and species distributions. Most of the field data and metadata are available through PPBio web sites or DataONE. Metadata is available for researchers that intend to explore the different faces of Brazilian biodiversity spatio-temporal variation, as well as for managers intending to improve conservation strategies. The Program also fostered, directly and indirectly, local technical capacity building, and supported the training of hundreds of undergraduate and graduate students. The main challenge is maintaining the long-term funding necessary to understand biodiversity patterns and processes under pressure from global environmental changes.

Background: Shortages and inequitable distribution of physicians is an obstacle to move towards Universal Health Coverage, especially in low-income and middle-income countries. In Brazil, expansion of medical school enrollment, curricula changes and recruitment programs were established to increase the number of physicians in underserved areas. This study seeks to analyze the impact of these measures in reduce inequities in access to medical education and physicians' distribution., Methods: This is an observational study that analyzes changes in the number of undergraduate medical places and number of physicians per inhabitants in different areas in Brazil between the years 2010 and 2018. Data regarding the number of undergraduate medical places, number and the practice location of physicians were obtained in public databases. Municipalities with less than 20,000 inhabitants were considered underserved areas. Data regarding access to antenatal visits were analyzed as a proxy for impact in access to healthcare., Results: From 2010 to 2018, 19,519 new medical undergraduate places were created which represents an increase of 120.2%. The increase in the number of physicians engaged in the workforce throughout the period was 113,702 physicians, 74,771 of these physicians in the Unified Health System. The greatest increase in the physicians per 1000 inhabitants ratio in the municipalities with the smallest population, the lowest Gross Domestic Product per capita and in those located in the states with the lowest concentration of physicians occurred in the 2013-2015 period. Increase in physician supply improved access to antenatal care., Conclusions: There was an expansion in the number of undergraduate medical places and medical workforce in all groups of municipalities assessed in Brazil. Medical undergraduate places expansion in the federal public schools was more efficient to reduce regional inequities in access to medical education than private sector expansion. The recruitment component of More Doctors for Brazil Program demonstrated effectiveness to increase the number of physicians in underserved areas. Our results indicate the importance of public policies to face inequities in access to medical education and physician shortages and the necessity of continuous assessment during the period of implementation, especially in the context of political and economic changes.