Threat directionality modulates defensive reactions in humans: cardiac and electrodermal responses.

Features of threatening cues and the associated context influence the perceived imminence of threat and the defensive responses evoked. To provide additional knowledge about how the directionality of a threat (i.e. directed-towards or away from the viewer) might impact defensive responses in humans, participants were shown pictures of a man carrying a gun (threat) or nonlethal object (neutral) directed-away from or towards the participant. Cardiac and electrodermal responses were collected. Compared to neutral images, threatening images depicting a gun directed-towards the participant induced sustained bradycardia and an increased electrodermal response, interpreted as immobility under attack. This defensive immobility reaction is evoked by high perceived threat and inescapable situations and indicates intense action preparation. Pictures of guns directed-away from the participant induced shorter bradycardia and no significant modulation of the electrodermal response compared to neutral pictures, which might be consistent with the perception of a less threatening situation. The results show that the directionality of threat stimuli is a key factor that prompts different patterns of defensive responses.

The antioxidant defence of Dekkera bruxellensis against hydrogen peroxide and its relationship to nitrate metabolism.

AIMS: The yeast Dekkera bruxellensis is a Crabtree-positive yeast that tends towards the oxidative/respiratory metabolism in aerobiosis. However, it is more sensitive to H2O2 than Saccharomyces cerevisiae. In order to investigate this metabolic paradox, the present work aimed to uncover the biological defence mechanism used by this yeast to tolerate the presence of exogenous H2O2. METHODS AND RESULTS: Growth curves and spot tests were performed to establish the values of minimal inhibitory concentration and minimal biocidal concentration of H2O2 for different combinations of carbon and nitrogen sources. Cells in exponential growth phase in different culture conditions were used to measure superoxide and thiols [protein (PT) and non-PT], enzyme activities and gene expression. CONCLUSIONS: The combination of glutathione peroxidase (Gpx) and sulfhydryl-containing PT formed the preferred defence mechanism against H2O2, which was more efficiently active under respiratory metabolism. However, the action of this mechanism was suppressed when the cells were metabolizing nitrate (NO3). SIGNIFICANCE AND IMPACT OF STUDY: These results were relevant to figure out the fitness of D. bruxellensis to metabolize industrial substrates containing oxidant molecules, such as molasses and plant hydrolysates, in the presence of a cheaper nitrogen source such as NO3.

Unstable environment of coastal lagoons drives genetic variation in the amphipod Quadrivisio lutzi.

The freshwater/brackish amphipod Quadrivisio lutzi inhabits coastal lagoons, highly unstable environments subject to sudden inflow of marine water. Our aim was to evaluate how the genetic composition varies in these populations. Brazilian populations were compared by 16S rRNA and COI gene sequences. The genetic structure of four Rio de Janeiro amphipod populations was evaluated during the period of 2011-2019 by COI. Rio de Janeiro population was compared with Alagoas and São Paulo populations, which was genetically distinct, at species level (16S, d > 7%; COI, d >14%). The genetic structure in Rio de Janeiro showed the Imboassica subpopulation as the most divergent (Imboassica & Carapebus, F ST = 0.238), followed by Lagamar population (Lagamar & Carapebus, F ST = 0.049). The geographic distance and urbanization around these lagoons explain the degree of genetic isolation of these amphipod subpopulations. Paulista and Carapebus populations were not structured. Temporal variation in haplotype number and frequency were evident in both populations that were evaluated (Carapebus and Imboassica). Changes in salinity and water volume variation at these lagoons may be responsible for the observed changes in genetic composition, which may be the results of genetic drift effects over temporally fluctuating size subpopulations, without loss of genetic diversity.

Dysphagia Perception Among Community-Dwelling Older Adults from a Municipality in Southern Brazil.

To analyse the prevalence of dysphagia perception and associated factors among community-dwelling older adults in Pelotas, Brazil. A total of 1447 community-dwelling individuals aged 60 and older participated in a cross-sectional population-based study carried out in 2014. Dysphagia perception was assessed using the following question: "Do you have swallowing difficulties? (Yes/No)". Independent variables included sociodemographic, behavioural and health characteristics. Poisson regression was used to obtain prevalence ratios. Dysphagia perception prevalence was 8.1%, higher among women (PR 1.63, 95% CI 1.07; 2.46) and in subjects older than 80 years (PR 1.88, 95% CI 1.16; 3.03). Older adults with 1-7 years of schooling were more likely to present dysphagia (PR 1.62; 95% CI 1.09; 2.40). Those who did not use dental prosthesis (PR 1.85; 95% CI 1.08; 3.16), who presented dry mouth sensation (PR 4.10; 95% CI 2.59; 6.51) and multimorbidity (PR 30.0; 95% CI 4.09; 219.45) were more likely to present dysphagia perception. The participants who consumed alcohol were 60% less likely to report dysphagia perception (PR 0.43; 95% CI 0.22; 0.86). One out of twelve older adults presented dysphagia perception, and associations with sociodemographic characteristics and other health problems were found. Early identification of dysphagia should be a public health and clinical concern.

Methylprednisolone as Adjunctive Therapy for Patients Hospitalized With Coronavirus Disease 2019 (COVID-19; Metcovid): A Randomized, Double-blind, Phase IIb, Placebo-controlled Trial.

BACKGROUND: Steroid use for coronavirus disease 2019 (COVID-19) is based on the possible role of these drugs in mitigating the inflammatory response, mainly in the lungs, triggered by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This study aimed to evaluate the efficacy of methylprednisolone (MP) among hospitalized patients with suspected COVID-19. METHODS: A parallel, double-blind, placebo-controlled, randomized, Phase IIb clinical trial was performed with hospitalized patients aged ≥18 years with clinical, epidemiological, and/or radiological suspected COVID-19 at a tertiary care facility in Manaus, Brazil. Patients were randomly allocated (1:1 ratio) to receive either intravenous MP (0.5 mg/kg) or placebo (saline solution) twice daily for 5 days. A modified intention-to-treat (mITT) analysis was conducted. The primary outcome was 28-day mortality. RESULTS: From 18 April to 16 June 2020, 647 patients were screened, 416 were randomized, and 393 were analyzed as mITT, with 194 individuals assigned to MP and 199 to placebo. SARS-CoV-2 infection was confirmed by reverse transcriptase polymerase chain reaction in 81.3%. The mortality rates at Day 28 were not different between groups. A subgroup analysis showed that patients over 60 years old in the MP group had a lower mortality rate at Day 28. Patients in the MP arm tended to need more insulin therapy, and no difference was seen in virus clearance in respiratory secretion until Day 7. CONCLUSIONS: The findings of this study suggest that a short course of MP in hospitalized patients with COVID-19 did not reduce mortality in the overall population. CLINICAL TRIALS REGISTRATION: NCT04343729.

Body mass index and mortality among community-dwelling elderly of Southern Brazil.

This study aims to measure the association between body mass index (BMI), comparing two different classifications, and mortality among community-dwelling elderly considering myopenia in Pelotas, Brazil. This is a longitudinal study started in 2014, we followed 1451 elderly people (≥ 60 years) enrolled in the "COMO VAI?" study. BMI was classified according to the World Health Organization (WHO) and the classification with specific cutoff points for older adults. Myopenia was measured by calf circumference (≤33 cm for women and ≤34 cm for men). Cox proportional-hazards models were used to test associations controlling for sociodemographic and behavioral characteristics and number of morbidities. Nearly 10% (N = 145) of the elderly died during almost three years of follow-up. We observed a L-shaped relation between BMI and mortality. Elderly with underweight had a higher mortality risk compared to those with adequate BMI in both classifications. According to the WHO classification, overweight elderly presented protection for mortality (HR: 0.58; 95% CI 0.38-0.87) when compared to those with adequate BMI. Among elderly with myopenia, overweight by WHO continued to protect against mortality, although not significantly, while those with the specific classification underweight presented a higher risk of death compared to those with normal weight (HR: 2.09; 95% CI 1.06-4.14). In conclusion the underweight increased the risk of death in community-dwelling elderly people during a follow-up of three years. The specific classification seemed to be more adequate to indicate risk of mortality in this population. Higher BMI protect against mortality when muscle mass was not considered.

Sleep duration trajectories from age 3 to 48 months in The Pelotas (Brazil) 2004 Birth Cohort Study.

BACKGROUND: Optimal sleep is essential for child growth, development, and immune function. Few studies have evaluated factors associated with sleep duration in childhood from a longitudinal perspective. OBJECTIVES: This study aimed to identify trajectories of sleep duration in childhood and associated maternal and child characteristics. METHODS: Sleep duration was assessed by maternal report at 3, 12, 24, and 48 months among children from the Pelotas (Brazil) 2004 Birth Cohort. Independent variables included family income, maternal and child demographics, and clinical characteristics. Trajectory analysis was carried out using a semi-parametric, group-based modelling approach. Multinomial logistic regression provided odds ratios (ORs) and 95% confidence intervals (CIs) for the associations between independent variables and sleep duration trajectory groups. RESULTS: A total of 3824 participants were included in the analyses. Three trajectories of sleep duration were identified: "short sleepers" (9.1%), "typical sleepers" (72.1%), and "initially longer sleepers" (18.8%). When compared to typical sleepers, children from less schooled mothers (OR 1.82, 95% CI 1.26, 2.62) and those whose mothers reported depressive symptoms during pregnancy (OR 1.31, 95% CI 1.02, 1.68) and consumed alcohol beverages at 3 months post-partum (OR 1.60, 95% CI 1.03, 2.50) were more likely to be short sleepers. Children who shared the bedroom with another child were about 40% (OR 1.41, 95% CI 1.07, 1.87) more likely to be short sleepers. None of the investigated maternal and child characteristics remained associated with the "initially longer sleeper" group. CONCLUSIONS: Among the identified trajectories, the group with short sleep duration trajectory deserves special attention given the importance of adequate sleep duration in the first years of life for the child's growth and development and the high concomitance of other risk factors, such as less schooled mothers, and mothers who reported depressive symptoms during pregnancy and consumed alcohol at 3 months post-partum.

Lawsone, a 2-hydroxy-1,4-naphthoquinone from Lawsonia inermis (henna), produces mitochondrial dysfunctions and triggers mitophagy in Saccharomyces cerevisiae.

Lawsone is a natural naphthoquinone present in the henna leaf extract with several cytotoxic activities and used as precursor for synthesis of various pharmaceutical compounds. Its biological activities are thought to be the result of oxidative stress generated, although the hydroxy group at position C-2 in its structure tends to reduce its electrophilic potential. In view of lack of knowledge on its activity, the present work aimed to elucidate the biological effect of lawsone using the yeast Saccharomyces cerevisiae. In the model strain BY4741 it was defined 229 mmol/L as the minimal inhibitory concentration (MIC). Using 172 mmol/L as sub-MIC value it was observed that yap1 deletion mutant was sensitive to lawsone independent the presence of oxygen. Lawsone affected yeast growth in glycerol, indicating interference in the respiratory metabolism. Intracellular content of thiol groups did not indicate intensive oxidative stress and the presence of the anti-oxidant N-acetylcysteine (NAC) exacerbated lawsone toxicity. By analysing the sensitivity of atg mutant strains and the localization of GFP-Atg8 fusion protein, it was concluded that lawsone primarily produces mitochondrial malfunctioning, leading to indirect oxidative stress. It triggers the autophagic response that ultimately induces mitophagy.

Manipulation of small Rho GTPases is a pathogen-induced process detected by NOD1.

Our innate immune system distinguishes microbes from self by detecting conserved pathogen-associated molecular patterns. However, these are produced by all microbes, regardless of their pathogenic potential. To distinguish virulent microbes from those with lower disease-causing potential the innate immune system detects conserved pathogen-induced processes, such as the presence of microbial products in the host cytosol, by mechanisms that are not fully resolved. Here we show that NOD1 senses cytosolic microbial products by monitoring the activation state of small Rho GTPases. Activation of RAC1 and CDC42 by bacterial delivery or ectopic expression of SopE, a virulence factor of the enteric pathogen Salmonella, triggered the NOD1 signalling pathway, with consequent RIP2 (also known as RIPK2)-mediated induction of NF-κB-dependent inflammatory responses. Similarly, activation of the NOD1 signalling pathway by peptidoglycan required RAC1 activity. Furthermore, constitutively active forms of RAC1, CDC42 and RHOA activated the NOD1 signalling pathway. Our data identify the activation of small Rho GTPases as a pathogen-induced process sensed through the NOD1 signalling pathway.

Lifelong robbery victimisation and mental disorders at age 18 years: Brazilian population-based study.

PURPOSE: Urban violence is a major problem in Brazil and may contribute to mental disorders among victims. The aim of this study was to assess the association between robbery victimisation and mental health disorders in late adolescence. METHODS: At age 18 years, 4106 participants in the 1993 Pelotas Birth Cohort Study were assessed. A questionnaire about history of robbery victimisation was administered, the Self-Report Questionnaire was used to screen for common mental disorders, and the Mini International Neuropsychiatric Interview was used to assess major depressive disorder and generalised anxiety disorder. Cross-sectional prevalence ratios between lifetime robbery victimisation and mental disorders were estimated using Poisson regression with robust standard errors, adjusting for socioeconomic variables measured at birth and violence in the home and maltreatment measured at age 15. RESULTS: There was a dose-response relationship between frequency of lifetime robberies and risk of mental disorders. Adolescents who had been robbed three or more times had twice the risk (PR 2.04; 95% CI 1.64-2.56) for common mental disorders, over four times the risk for depression (PR 4.59; 95% CI 2.60-8.12), and twice the risk for anxiety (PR 1.93; 95% CI 1.06-3.50), compared with non-victims, adjusting for covariates. Experiencing frequent robberies had greater impact on common mental disorders than experiencing an armed robbery. Population attributable fractions with regard to robbery were 9% for common mental disorders, 13% for depression, and 8% for anxiety. CONCLUSIONS: Robberies are associated with common mental disorders in late adolescence, independently of violence between family members. Reducing urban violence could significantly help in preventing common mental illnesses.

Ascorbate peroxidase overexpression protects Leishmania braziliensis against trivalent antimony effects.

Ascorbate peroxidase (APX) is a redox enzyme of the trypanothione pathway that converts hydrogen peroxide (H2O2) into water molecules. In the present study, the APX gene was overexpressed in Leishmania braziliensis to investigate its contribution to the trivalent antimony (SbIII)-resistance phenotype. Western blot results demonstrated that APX-overexpressing parasites had higher APX protein levels in comparison with the wild-type line (LbWTS). APX-overexpressing clones showed an 8-fold increase in the antimony-resistance index over the parental line. In addition, our results indicated that these clones were approximately 1.8-fold more tolerant to H2O2 than the LbWTS line, suggesting that the APX enzyme plays an important role in the defence against oxidative stress. Susceptibility tests revealed that APX-overexpressing L. braziliensis lines were more resistant to isoniazid, an antibacterial agent that interacts with APX. Interestingly, this compound enhanced the anti-leishmanial SbIII effect, indicating that this combination represents a good strategy for leishmaniasis chemotherapy. Our data demonstrate that APX enzyme is involved in the development of L. braziliensis antimony-resistance phenotype and may be an attractive therapeutic target in the design of new strategies for leishmaniasis treatment.

Supervised machine learning techniques to predict binding affinity. A study for cyclin-dependent kinase 2.

Here we report the development of a machine-learning model to predict binding affinity based on the crystallographic structures of protein-ligand complexes. We used an ensemble of crystallographic structures (resolution better than 1.5 Å resolution) for which half-maximal inhibitory concentration (IC50) data is available. Polynomial scoring functions were built using as explanatory variables the energy terms present in the MolDock and PLANTS scoring functions. Prediction performance was tested and the supervised machine learning models showed improvement in the prediction power, when compared with PLANTS and MolDock scoring functions. In addition, the machine-learning model was applied to predict binding affinity of CDK2, which showed a better performance when compared with AutoDock4, AutoDock Vina, MolDock, and PLANTS scores.

Malaria parasite infection compromises control of concurrent systemic non-typhoidal Salmonella infection via IL-10-mediated alteration of myeloid cell function.

Non-typhoidal Salmonella serotypes (NTS) cause a self-limited gastroenteritis in immunocompetent individuals, while children with severe Plasmodium falciparum malaria can develop a life-threatening disseminated infection. This co-infection is a major source of child mortality in sub-Saharan Africa. However, the mechanisms by which malaria contributes to increased risk of NTS bacteremia are incompletely understood. Here, we report that in a mouse co-infection model, malaria parasite infection blunts inflammatory responses to NTS, leading to decreased inflammatory pathology and increased systemic bacterial colonization. Blunting of NTS-induced inflammatory responses required induction of IL-10 by the parasites. In the absence of malaria parasite infection, administration of recombinant IL-10 together with induction of anemia had an additive effect on systemic bacterial colonization. Mice that were conditionally deficient for either myeloid cell IL-10 production or myeloid cell expression of IL-10 receptor were better able to control systemic Salmonella infection, suggesting that phagocytic cells are both producers and targets of malaria parasite-induced IL-10. Thus, IL-10 produced during the immune response to malaria increases susceptibility to disseminated NTS infection by suppressing the ability of myeloid cells, most likely macrophages, to control bacterial infection.

Interactions of the human pathogenic Brucella species with their hosts.

Brucellosis is a zoonotic infection caused primarily by the bacterial pathogens Brucella melitensis and B. abortus. It is acquired by consumption of unpasteurized dairy products or by contact with infected animals. Globally, it is one of the most widespread zoonoses, with 500,000 new cases reported each year. In endemic areas, Brucella infections represent a serious public health problem that results in significant morbidity and economic losses. An important feature of the disease is persistent bacterial colonization of the reticuloendothelial system. In this review we discuss recent insights into mechanisms of intracellular survival and immune evasion that contribute to systemic persistence by the pathogenic Brucella species.

CD4+ T cell-derived IL-10 promotes Brucella abortus persistence via modulation of macrophage function.

Evasion of host immune responses is a prerequisite for chronic bacterial diseases; however, the underlying mechanisms are not fully understood. Here, we show that the persistent intracellular pathogen Brucella abortus prevents immune activation of macrophages by inducing CD4(+)CD25(+) T cells to produce the anti-inflammatory cytokine interleukin-10 (IL-10) early during infection. IL-10 receptor (IL-10R) blockage in macrophages resulted in significantly higher NF-kB activation as well as decreased bacterial intracellular survival associated with an inability of B. abortus to escape the late endosome compartment in vitro. Moreover, either a lack of IL-10 production by T cells or a lack of macrophage responsiveness to this cytokine resulted in an increased ability of mice to control B. abortus infection, while inducing elevated production of pro-inflammatory cytokines, which led to severe pathology in liver and spleen of infected mice. Collectively, our results suggest that early IL-10 production by CD25(+)CD4(+) T cells modulates macrophage function and contributes to an initial balance between pro-inflammatory and anti-inflammatory cytokines that is beneficial to the pathogen, thereby promoting enhanced bacterial survival and persistent infection.

Salmonella uses energy taxis to benefit from intestinal inflammation.

Chemotaxis enhances the fitness of Salmonella enterica serotype Typhimurium (S. Typhimurium) during colitis. However, the chemotaxis receptors conferring this fitness advantage and their cognate signals generated during inflammation remain unknown. Here we identify respiratory electron acceptors that are generated in the intestinal lumen as by-products of the host inflammatory response as in vivo signals for methyl-accepting chemotaxis proteins (MCPs). Three MCPs, including Trg, Tsr and Aer, enhanced the fitness of S. Typhimurium in a mouse colitis model. Aer mediated chemotaxis towards electron acceptors (energy taxis) in vitro and required tetrathionate respiration to confer a fitness advantage in vivo. Tsr mediated energy taxis towards nitrate but not towards tetrathionate in vitro and required nitrate respiration to confer a fitness advantage in vivo. These data suggest that the energy taxis receptors Tsr and Aer respond to distinct in vivo signals to confer a fitness advantage upon S. Typhimurium during inflammation by enabling this facultative anaerobic pathogen to seek out favorable spatial niches containing host-derived electron acceptors that boost its luminal growth.

Neutrophils are a source of gamma interferon during acute Salmonella enterica serovar Typhimurium colitis.

Gamma interferon (IFN-γ) is an important driver of intestinal inflammation during colitis caused by Salmonella enterica serovar Typhimurium. Here we used the mouse colitis model to investigate the cellular sources of IFN-γ in the cecal mucosa during the acute phase of an S. Typhimurium infection. While IFN-γ staining was detected in T cells, NK cells, and inflammatory monocytes at 2 days after infection, the majority of IFN-γ-positive cells in the cecal mucosa were neutrophils. Furthermore, neutrophil depletion blunted mucosal Ifng expression and reduced the severity of intestinal lesions during S. Typhimurium infection. We conclude that neutrophils are a prominent cellular source of IFN-γ during the innate phase of S. Typhimurium-induced colitis.

Very long O-antigen chains enhance fitness during Salmonella-induced colitis by increasing bile resistance.

Intestinal inflammation changes the luminal habitat for microbes through mechanisms that have not been fully resolved. We noticed that the FepE regulator of very long O-antigen chain assembly in the enteric pathogen Salmonella enterica serotype Typhimurium (S. Typhimurium) conferred a luminal fitness advantage in the mouse colitis model. However, a fepE mutant was not defective for survival in tissue, resistance to complement or resistance to polymyxin B. We performed metabolite profiling to identify changes in the luminal habitat that accompany S. Typhimurium-induced colitis. This analysis suggested that S. Typhimurium-induced colitis increased the luminal concentrations of total bile acids. A mutation in fepE significantly reduced the minimal inhibitory concentration (MIC) of S. Typhimurium for bile acids in vitro. Oral administration of the bile acid sequestrant cholestyramine resin lowered the concentrations of total bile acids in colon contents during S. Typhimurium infection and significantly reduced the luminal fitness advantage conferred by the fepE gene in the mouse colitis model. Collectively, these data suggested that very long O-antigen chains function in bile acid resistance of S. Typhimurium, a property conferring a fitness advantage during luminal growth in the inflamed intestine.

Innate immune recognition of flagellin limits systemic persistence of Brucella.

Brucella are facultative intracellular bacteria that cause chronic infections by limiting innate immune recognition. It is currently unknown whether Brucella FliC flagellin, the monomeric subunit of flagellar filament, is sensed by the host during infection. Here, we used two mutants of Brucella melitensis, either lacking or overexpressing flagellin, to show that FliC hinders bacterial replication in vivo. The use of cells and mice genetically deficient for different components of inflammasomes suggested that FliC was a target of the cytosolic innate immune receptor NLRC4 in vivo but not in macrophages in vitro where the response to FliC was nevertheless dependent on the cytosolic adaptor ASC, therefore suggesting a new pathway of cytosolic flagellin sensing. However, our work also suggested that the lack of TLR5 activity of Brucella flagellin and the regulation of its synthesis and/or delivery into host cells are both part of the stealthy strategy of Brucella towards the innate immune system. Nevertheless, as a flagellin-deficient mutant of B. melitensis wasfound to cause histologically demonstrable injuries in the spleen of infected mice, we suggested that recognition of FliC plays a role in the immunological stand-off between Brucella and its host, which is characterized by a persistent infection with limited inflammatory pathology.