Ancestral aneuploidy and stable chromosomal duplication resulting in differential genome structure and gene expression control in trypanosomatid parasites.

Aneuploidy is widely observed in both unicellular and multicellular eukaryotes, usually associated with adaptation to stress conditions. Chromosomal duplication stability is a tradeoff between the fitness cost of having unbalanced gene copies and the potential fitness gained from increased dosage of specific advantageous genes. Trypanosomatids, a family of protozoans that include species that cause neglected tropical diseases, are a relevant group to study aneuploidies. Their life cycle has several stressors that could select for different patterns of chromosomal duplications and/or losses, and their nearly universal use of polycistronic transcription increases their reliance on gene expansion/contraction, as well as post-transcriptional control as mechanisms for gene expression regulation. By evaluating the data from 866 isolates covering seven trypanosomatid genera, we have revealed that aneuploidy tolerance is an ancestral characteristic of trypanosomatids but has a reduced occurrence in a specific monophyletic clade that has undergone large genomic reorganization and chromosomal fusions. We have also identified an ancient chromosomal duplication that was maintained across these parasite's speciation, named collectively as the trypanosomatid ancestral supernumerary chromosome (TASC). TASC has most genes in the same coding strand, is expressed as a disomic chromosome (even having four copies), and has increased potential for functional variation, but it purges highly deleterious mutations more efficiently than other chromosomes. The evidence of stringent control over gene expression in this chromosome suggests that these parasites have adapted to mitigate the fitness cost associated with this ancient chromosomal duplication.

Complete assembly, annotation of virulence genes and CRISPR editing of the genome of Leishmania amazonensis PH8 strain.

We report the sequencing and assembly of the PH8 strain of Leishmania amazonensis one of the etiological agents of leishmaniasis. After combining data from long Pacbio reads, short Illumina reads and synteny with the Leishmania mexicana genome, the sequence of 34 chromosomes with 8317 annotated genes was generated. Multigene families encoding three virulence factors, A2, amastins and the GP63 metalloproteases, were identified and compared to their annotation in other Leishmania species. As they have been recently recognized as virulence factors essential for disease establishment and progression of the infection, we also identified 14 genes encoding proteins involved in parasite iron and heme metabolism and compared to genes from other Trypanosomatids. To follow these studies with a genetic approach to address the role of virulence factors, we tested two CRISPR-Cas9 protocols to generate L. amazonensis knockout cell lines, using the Miltefosine transporter gene as a proof of concept.

How is smoking distributed in relation to socioeconomic status? Evidence from Brazil in the years 2013 and 2019.

The present study aimed to analyze income-related inequality in tobacco consumption in Brazil using data from the National Health Survey at two points in time (2013 and 2019). This study contributes to the growing literature analyzing socioeconomic inequalities in tobacco use by investigating income-related inequalities in the consumption of different tobacco products in Brazil. The inequality measure is the concentration index with an Erreygers correction (EI), and the analysis of its decomposition allows the identification of the factors that determine such inequality. There is inequality in smoking concentrated in the poorest persons, and this pattern also occurs for manufactured cigarettes and roll-your-own cigarettes (RYO), while inequality in smoking cessation is concentrated among the wealthiest. Smoking inequalities were greater in men, older age groups, and RYO. In terms of evolution, the overall results indicated a small decline in smoking inequality. For the decomposition analysis, the results show that the main factors that affect tobacco inequality in terms of concentration in the poorest are education, income, and having private health insurance. The region variable, by contrast, has a positive contribution, since the wealthiest regions have individuals who are more likely to smoke. These results have important implications that serve as a basis for formulating public health policies. For example, greater inequalities for men and older individuals can be targeted by public policies with a special focus on these cases.

Bacteria, yeasts, and fungi associated with larval food of Brazilian native stingless bees.

Stingless bees are a diverse group with a relevant role in pollinating native species. Its diet is rich in carbohydrates and proteins, by collecting pollen and nectar supplies the development of its offspring. Fermentation of these products is associated with microorganisms in the colony. However, the composition of microorganisms that comprise this microbiome and its fundamental role in colony development is still unclear. To characterize the colonizing microorganisms of larval food in the brood cells of stingless bees Frieseomelitta varia, Melipona quadrifasciata, Melipona scutellaris, and Tetragonisca angustula, we have utilized molecular and culture-based techniques. Bacteria of the phyla Firmicutes, Proteobacteria, Actinobacteria, and fungi of the phyla Ascomycota, Basidiomycota, Mucoromycota, and Mortierellomycota were found. Diversity analysis showed that F. varia had a greater diversity of bacteria in its microbiota, and T. angustula had a greater diversity of fungi. The isolation technique allowed the identification of 189 bacteria and 75 fungi. In summary, this research showed bacteria and fungi associated with the species F. varia, M. quadrifasciata, M. scutellaris, and T. angustula, which may play an essential role in the survival of these organisms. Besides that, a biobank with bacteria and fungus isolates from LF of Brazilian stingless bees was created, which can be used for different studies and the prospection of biotechnology compounds.

Molecular and biochemical responses of sesame (Sesame indicum L.) to rhizobacteria inoculation under water deficit.

Introduction: Water scarcity is a challenge for sesame cultivation under rainfed conditions. In this scenario, a potential strategy to alleviate the water deficit is the application of plant growth-promoting bacteria. The objective of this study was to analyze the interaction of rhizobacteria with sesame cultivation under water deficit conditions. Methods: An experiment was conducted in pots in a greenhouse using the BRS Morena sesame cultivar. The experimental design was completely randomized in a factorial scheme: 2 (irrigation regimes - daily irrigation and water deficit by suspending irrigation until 90% stomatal closure) x 6 (treatments with nitrogen or inoculants), with 5 replications. The types of fertilization were characterized by the addition of nitrogen (ammonium sulfate; 21% N), inoculants based on Bacillus spp. (pant001, ESA 13, and ESA 402), Agrobacterium sp. (ESA 441), and without nitrogen (control). On the fifth day after the suspension of irrigation, plant material was collected for gene expression analysis (DREB1 and HDZ7), activities of antioxidant enzymes (superoxide dismutase and catalase), relative proline content, and photosynthetic pigments. At the end of the crop cycle (about 85 days), production characteristics (root dry matter, aboveground dry matter, number of capsules, and thousand seed weight), as well as leaf nitrogen (N) and phosphorus (P) content, were evaluated. Results and Discussion: There was a positive effect on both production and biochemical characteristics (proline, superoxide dismutase, catalase, and photosynthetic pigments). Regarding gene expression, most of the inoculated treatments exhibited increased expression of the DREB1 and HDZ7 genes. These biological indicators demonstrate the potential of rhizobacteria for application in sesame cultivation, providing nutritional supply and reducing the effects of water deficit.

Temperature and pressure dependent rate constants of the reactions of OH• with cyclopentene from variational TST and SS-QRRK methods.

In this work, the pressure- and temperature-dependent reaction rate constants for the hydrogen abstraction and addition of hydroxyl radicals to the unsaturated cyclopentene were studied. Geometries and vibrational frequencies of reactants, products, and transition states were calculated using density functional theory, with single-point energy corrections determined at the domain-based local pair natural orbital-coupled-cluster single double triple/cc-pVTZ-F12 level. The high-pressure limit rate constants were calculated using the canonical variational transition state theory with the small-curvature tunneling approximation. The vibrational partition functions were corrected by the effects of torsional and ring-puckering anharmonicities of the transition states and cyclopentene, respectively. Variational effects are shown to be relevant for all the hydrogen abstraction reactions. The increasing of the rate constants by tunneling is significant at temperatures below 500 K. The pressure dependence on the rate constants of the addition of OH• to cyclopentene was calculated using the system-specific quantum Rice-Ramsperger-Kassel model. The high-pressure limit rate constants decrease with increasing temperature in the range 250-1000 K. The falloff behavior was studied at several temperatures with pressures varying between 10-3 and 103 bar. At temperatures below 500 K, the effect of the pressure on the addition rate constant is very modest. However, at temperatures around and above 1000 K, taking pressure into account is mandatory for an accurate rate constant calculation. Branching ratio analyses reveal that the addition reaction dominates at temperatures below 500 K, decreasing rapidly at higher temperatures. Arrhenius parameters are provided for all reactions and pressure dependent Arrhenius parameters are given for the addition of OH• to cyclopentene.

Draft Genome Sequence of the Protozoan Parasite Leishmania braziliensis Strain BA788, Isolated from a Clinical Case in Bahia State, Brazil.

The draft genome of the parasite Leishmania braziliensis strain BA788, which was isolated from a patient from Bahia state, Brazil, was sequenced using Illumina paired-end technology. The assembled genome is 33.5 Mb long and contains 7,603 genes. This genome will contribute to studies aimed at understanding the pathogenesis caused by this parasite strain.

Accessing the Variability of Multicopy Genes in Complex Genomes using Unassembled Next-Generation Sequencing Reads: The Case of Trypanosoma cruzi Multigene Families.

Repetitive elements cause assembly fragmentation in complex eukaryotic genomes, limiting the study of their variability. The genome of Trypanosoma cruzi, the parasite that causes Chagas disease, has a high repetitive content, including multigene families. Although many T. cruzi multigene families encode surface proteins that play pivotal roles in host-parasite interactions, their variability is currently underestimated, as their high repetitive content results in collapsed gene variants. To estimate sequence variability and copy number variation of multigene families, we developed a read-based approach that is independent of gene-specific read mapping and de novo assembly. This methodology was used to estimate the copy number and variability of MASP, TcMUC, and Trans-Sialidase (TS), the three largest T. cruzi multigene families, in 36 strains, including members of all six parasite discrete typing units (DTUs). We found that these three families present a specific pattern of variability and copy number among the distinct parasite DTUs. Inter-DTU hybrid strains presented a higher variability of these families, suggesting that maintaining a larger content of their members could be advantageous. In addition, in a chronic murine model and chronic Chagasic human patients, the immune response was focused on TS antigens, suggesting that targeting TS conserved sequences could be a potential avenue to improve diagnosis and vaccine design against Chagas disease. Finally, the proposed approach can be applied to study multicopy genes in any organism, opening new avenues to access sequence variability in complex genomes. IMPORTANCE Sequences that have several copies in a genome, such as multicopy-gene families, mobile elements, and microsatellites, are among the most challenging genomic segments to study. They are frequently underestimated in genome assemblies, hampering the correct assessment of these important players in genome evolution and adaptation. Here, we developed a new methodology to estimate variability and copy numbers of repetitive genomic regions and employed it to characterize the T. cruzi multigene families MASP, TcMUC, and transsialidase (TS), which are important virulence factors in this parasite. We showed that multigene families vary in sequence and content among the parasite's lineages, whereas hybrid strains have a higher sequence variability that could be advantageous to the parasite's survivability. By identifying conserved sequences within multigene families, we showed that the mammalian host immune response toward these multigene families is usually focused on the TS multigene family. These TS conserved and immunogenic peptides can be explored in future works as diagnostic targets or vaccine candidates for Chagas disease. Finally, this methodology can be easily applied to any organism of interest, which will aid in our understanding of complex genomic regions.

Quality of Life in Community-Dwelling Older People with Functional and Nutritional Impairment and Depressive Symptoms: A Comparative Cross-Sectional Study in Brazil and Portugal.

BACKGROUND: The interaction of quality of life (QoL) with functionality, nutrition and depression has been studied, but few studies have compared different realities. Our objective was to compare the associations of QoL with impaired functionality, nutritional status and depressive symptoms among older people patients treated in primary health care (PHC) in Brazil and Portugal. METHODS: Cross-sectional, comparative study was conducted with primary data from PHC services in Brazil and Portugal with users over 65 years old. Participants' scores were classified as "impaired" and "preserved" for QoL, functional decline, nutrition and depression. We used Pearson's chi-square test, Fisher's exact test and the Mann-Whitney U test. RESULTS: Our sample had a total of 150 PHC users. We found lower QoL scores in Brazil, which were associated with the risk of functional decline for the domains Physical Functioning, General Health Perceptions, Mental Health dimensions and Physical Health. Nutritional impairment in the group from Portugal included the domains of Vitality and Social Role Functioning. For depressive impairment, Portugal showed an association with the domains Mental Health, Vitality and Social Role Functioning. CONCLUSIONS: QoL was associated with functional and nutritional impairment and depressive symptoms, highlighting physical, mental and social characteristics related to the perception of well-being.

Investigation of pre and post environmental impact of the lockdown (COVID-19) on the water quality of the Capibaribe and Tejipió rivers, Recife metropolitan region, Brazil.

The coronavirus pandemic has seriously affected human health, although some improvements on environmental indexes have temporarily occurred, due to changes on socio-cultural and economic standards. The objective of this study was to evaluate the impacts of the coronavirus and the influence of the lockdown associated with rainfall on the water quality of the Capibaribe and Tejipió rivers, Recife, Northeast Brazil, using cloud remote sensing on the Google Earth Engine (GEE) platform. The study was carried out based on eight representative images from Sentinel-2. Among the selected images, two refer to the year 2019 (before the pandemic), three refer to 2020 (during a pandemic), two from the lockdown period (2020), and one for the year 2021. The land use and land cover (LULC) and slope of the study region were determined and classified. Water turbidity data were subjected to descriptive and multivariate statistics. When analyzing the data on LULC for the riparian margin of the Capibaribe and Tejipió rivers, a low permanent preservation area was found, with a predominance of almost 100% of the urban area to which the deposition of soil particles in rivers are minimal. The results indicated that turbidity values in the water bodies varied from 6 mg. L-1 up to 40 mg. L-1. Overall, the reduction in human-based activities generated by the lockdown enabled improvements in water quality of these urban rivers.

Replacement of Leishmania (Leishmania) infantum Populations in an Endemic Focus of Visceral Leishmaniasis in Brazil.

Visceral leishmaniasis is an important global health problem with an estimated of 50,000 to 90,000 new cases per year. VL is the most serious form of leishmaniasis as it can be fatal in 95% of the cases if it remains untreated. VL is a particularly acute problem in Brazil which contributed with 97% of all cases reported in 2020 in the Americas. In this country, VL affects mainly the poorest people in both urban and rural areas and continues to have a high mortality rate estimated around 8.15%. Here, we performed a temporal parasite population study using whole genome sequence data from a set of 34 canine isolates sampled in 2008, 2012 and 2015 from a re-emergent focus in Southeastern Brazil. Our study found the presence of two distinct sexual subpopulations that corresponded to two isolation periods. These subpopulations diverged hundreds of years ago with no apparent gene flow between them suggesting a process of rapid replacement during a two-year period. Sequence comparisons and analysis of nucleotide diversity also showed evidence of balancing selection acting on transport-related genes and antigenic families. To our knowledge this is the first population genomic study showing a turn-over of parasite populations in an endemic region for leishmaniasis. The complexity and rapid adaptability of these parasites pose new challenges to control activities and demand more integrated approaches to understand this disease in New World foci.

Trypanosoma cruzi iron superoxide dismutases: insights from phylogenetics to chemotherapeutic target assessment.

BACKGROUND: Components of the antioxidant defense system in Trypanosoma cruzi are potential targets for new drug development. Superoxide dismutases (SODs) constitute key components of antioxidant defense systems, removing excess superoxide anions by converting them into oxygen and hydrogen peroxide. The main goal of the present study was to investigate the genes coding for iron superoxide dismutase (FeSOD) in T. cruzi strains from an evolutionary perspective. METHODS: In this study, molecular biology methods and phylogenetic studies were combined with drug assays. The FeSOD-A and FeSOD-B genes of 35 T. cruzi strains, belonging to six discrete typing units (Tcl-TcVI), from different hosts and geographical regions were amplified by PCR and sequenced using the Sanger method. Evolutionary trees were reconstructed based on Bayesian inference and maximum likelihood methods. Drugs that potentially interacted with T. cruzi FeSODs were identified and tested against the parasites. RESULTS: Our results suggest that T. cruzi FeSOD types are members of distinct families. Gene copies of FeSOD-A (n = 2), FeSOD-B (n = 4) and FeSOD-C (n = 4) were identified in the genome of the T. cruzi reference clone CL Brener. Phylogenetic inference supported the presence of two functional variants of each FeSOD type across the T. cruzi strains. Phylogenetic trees revealed a monophyletic group of FeSOD genes of T. cruzi TcIV strains in both distinct genes. Altogether, our results support the hypothesis that gene duplication followed by divergence shaped the evolution of T. cruzi FeSODs. Two drugs, mangafodipir and polaprezinc, that potentially interact with T. cruzi FeSODs were identified and tested in vitro against amastigotes and trypomastigotes: mangafodipir had a low trypanocidal effect and polaprezinc was inactive. CONCLUSIONS: Our study contributes to a better understanding of the molecular biodiversity of T. cruzi FeSODs. Herein we provide a successful approach to the study of gene/protein families as potential drug targets.

Antigenic diversity of MASP gene family of Trypanosoma cruzi.

Trypanosoma cruzi, the etiological agent of Chagas disease (CD), is a heterogeneous species with high genetic and phenotypic diversity. MASP is the second largest multigene family of T. cruzi. The high degree of polymorphism of the family associated with its location at the surface of infective forms of T. cruzi suggests that MASP participates in mechanisms of host-parasite interaction. In this work, MASP members were divided into 7 subgroups based on protein sequence similarity, and one representative member from each subgroup was chosen to be expressed recombinantly. Immunogenicity of recombinant MASP proteins (rMASP) was investigated using different sera panels from T. cruzi infected mice. To mimic a natural condition in which different MASP members are expressed at the same time in the parasite population, a multiplex bead-based flow cytometry assay was also standardized. Results showed that rMASPs are poorly recognized by sera from mice infected with Colombiana strain, whereas sera from mice infected with CL Brener and Y display high reactivity against the majority of rMASPs tested. Flow cytometry showed that MASP recognition profile changes 10 days after infection. Also, multiplex assay suggests that MASP M1 and M2 are more immunogenic than the other MASP members evaluated that may play an immunodominant role during infection.

Multiplex PCR assay for correct identification of the fish pathogenic species of Edwardsiella genus reveals the presence of E. anguillarum in South America in strains previously characterized as E. tarda.

AIMS: Develop a species-specific multiplex PCR to correctly identify Edwardsiella species in routine diagnostic for fish bacterial diseases. METHODS AND RESULTS: The genomes of 62 Edwardsiella spp. isolates available from the National Center for Biotechnology Information (NCBI) database were subjected to taxonomic and pan-genomic analyses to identify unique regions that could be exploited by species-specific PCR. The designed primers were tested against isolated Edwardsiella spp. strains, revealing errors in commercial biochemical tests for bacterial classification regarding Edwardsiella species. CONCLUSION: Some of the genomes of Edwardsiella spp. in the NCBI platform were incorrectly classified, which can lead to errors in some research. A functional mPCR was developed to differentiate between phenotypically and genetically ambiguous Edwardsiella, with which, we detected the presence of Edwardsiella anguillarum affecting fish in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that the misclassification of Edwardsiella spp in Brazil concealed the presence of E. anguillarum in South America. Also, this review of the taxonomic classification of the Edwardsiella genus is a contribution to the field to help researchers with their sequencing and identification of genomes, showing some misclassifications in online databases that must be corrected, as well as developing an easy assay to characterize Edwardsiella species in an end-point mPCR.

Disruption of Active Trans-Sialidase Genes Impairs Egress from Mammalian Host Cells and Generates Highly Attenuated Trypanosoma cruzi Parasites.

Trans-sialidases (TS) are unusual enzymes present on the surface of Trypanosoma cruzi, the causative agent of Chagas disease. Encoded by the largest gene family in the T. cruzi genome, only few members of the TS family have catalytic activity. Active trans-sialidases (aTS) are responsible for transferring sialic acid from host glycoconjugates to mucins, also present on the parasite surface. The existence of several copies of TS genes has impaired the use of reverse genetics to study this highly polymorphic gene family. Using CRISPR-Cas9, we generated aTS knockout cell lines displaying undetectable levels of TS activity, as shown by sialylation assays and labeling with antibodies that recognize sialic acid-containing mucins. In vitro infection assays showed that disruption of aTS genes does not affect the parasite's capacity to invade cells or to escape from the parasitophorous vacuole but resulted in impaired differentiation of amastigotes into trypomastigotes and parasite egress from the cell. When inoculated into mice, aTS mutants were unable to establish infection even in the highly susceptible gamma interferon (IFN-γ) knockout mice. Mice immunized with aTS mutants were fully protected against a challenge infection with the virulent T. cruzi Y strain. Altogether, our results confirmed the role of aTS as a T. cruzi virulence factor and indicated that aTS play a major role during the late stages of intracellular development and parasite egress. Notably, mutants lacking TS activity are completely avirulent in animal models of infection and may be used as a live attenuated vaccine against Chagas disease. IMPORTANCE Trypanosoma cruzi is the causative agent of Chagas disease, a neglected tropical disease that affects approximately 6 to 8 million people and for which there is no effective treatment or vaccine. The parasite expresses a family of surface proteins, named trans-sialidases, responsible for transferring sialic acid from host glycoconjugates to parasite mucins. Although recognized as a main virulence factor, the multiple roles of these proteins during infection have not yet been fully characterized, mainly because the presence of several copies of aTS genes has impaired their study using reverse genetics. By applying CRISPR-Cas9, we generated aTS knockout parasites and showed that, although aTS parasite mutants were able to infect cells in vitro, they have an impaired capacity to egress from the infected cell. Importantly, aTS mutants lost the ability to cause infection in vivo but provided full protection against a challenge infection with a virulent strain.

Anatomical Risk Factors for Anterior Cruciate Ligament Injury Are Not Important As Patellar Instability Risk Factors in Patients with Acute Knee Injury.

To compare in magnetic resonance imaging the anatomical risk factors for anterior cruciate ligament (ACL) injury and patellar dislocation among patients who suffered acute knee injury, 105 patients with acute knee injury resulting in 38 patellar dislocations (patella group), 35 ACL injuries (ACL group), and 32 meniscus or medial collateral ligament injuries (control group) were included. These groups were compared for risk factors for patellar dislocation (patellar height, trochlear dysplasia, and quadriceps angle of action) and for ACL injury (intercondylar width, posterior inclination of tibial plateaus, and depth of the medial plateau). Univariate analysis found statistically significant differences (p < 0.05) between the patella and ACL groups in patellar height (Caton-Deschamps [CD] 1.23 vs. 1.07), trochlear facet asymmetry (55 vs. 68%), PTTG (13.08 vs. 8.01 mm), and the patellar tip and trochlear groove (PTTG) angle (29.5 vs. 13.71 degrees). The patella group also differed from control in medial plateau inclination (4.8 vs. 1.87 degrees), patellar height (CD 1.23 vs 1.08), trochlear facet asymmetry (55 vs. 69%), lateral trochlear inclination (17.11 vs. 20.65 degrees), trochlear depth (4.1 vs. 6.05 mm), PTTG (13.08 vs. 9.85 mm), and the PTTG angle (29.5 vs. 17.88 degrees). The ACL and control groups were similar in all measures. Multivariate analysis found the following significant determinants between the Patella and Control groups: patellar height (CD index, odds ratio [OR]: 80.13, p = 0.015), trochlear anatomy (asymmetry of facets M/L, OR: 1.06, p = 0.031) and quadriceps action angle (PTTG angle, OR: 1.09, p = 0.016); between the ACL and control groups: PTTG angle (OR: 0.936, p = 0.04) and female gender (OR: 3.876, p = 0.032); and between the patella and ACL groups, the CD index (OR: 67.62, p = 0.026), asymmetry of the M/L facets (OR: 1.07, p = 0.011) and PTTG angle (OR: 1.16, p < 0.001). In conclusion, in patients with acute knee injury, the anatomical factors patellar height, trochlear dysplasia, and quadriceps angle of action were related to the occurrence of patellar dislocation. None of the anatomical factors studied was related to the occurrence of anterior cruciate ligament injury.

Growth performance and rumen morphometrics of Nellore and ½ Angus/Nellore feedlot cattle adapted over 9 and 14 days to high-concentrate diets.

The objective of this study was to evaluate the effects of adapting Nellore and ½ Angus/Nellore (AN) feedlot cattle over periods of 9 and 14 days to high-concentrate diets on performance, feeding behaviour, carcass traits and rumen morphometrics. Seventy-two yearling bulls (313.5 kg ± 24.5), 36 Nellore and 36 AN, were randomly allocated in 24 pens (3 animals/pen; 24 m2 and 2.0 m of bunk space/animal) according to a randomized complete block design with a 2 × 2 factorial arrangement of treatments as follows: Nellore adapted for 9 days, Nellore adapted for 14 days, AN adapted for 9 days, and AN adapted for 14 days. Each treatment was composed by 6 pens (considered the experimental unit in this study). The adaptation lasted either 9 or 14 days and consisted of 3 step-up diets. Therefore, yearling bulls received the finishing diet containing 86% concentrate either on day 10 or 15 of the study, which lasted 89 days taking into account adaptation and finishing periods. Cattle were slaughtered in a commercial abattoir, and two 1-cm2 -rumen fragments, one from cranial and another from ventral sac, were collected. The AN cattle outperformed Nellore in terms of average daily gain (1.71 kg/day vs. 1.27 kg/day, p < 0.01), gain:feed ratio (0.137 kg/kg vs. 0.127 kg/kg, p = 0.02) and hot carcass weight (243.64 kg vs. 228.98 kg, p < 0.01). No main effect of the adaptation period was observed for any of the feedlot performance and carcass traits variables evaluated. Compared to feedlot cattle adapted for 9 days, feedlot cattle adapted for 14 days sorted against long (0.68 vs. 0.91, p < 0.01) and for fine particles (1.04 vs. 1.00, p = 0.01). An interaction (p < 0.01) of genotype and adaptation period was observed for rumenitis, where Nellore bulls adapted for 14 days presented the highest scores. In conclusion, there was no evidence that either Nellore or AN cattle benefit from an adaptation period shorter than 14 days.

Socioeconomic inequality in health in older adults in Brazil.

Objective: This study analyzed socioeconomic inequality in self-rated health for older adults (aged fifty or over) in Brazil. Methods: Data from the 2015-2016 Brazilian Longitudinal Study of Aging (ELSI-Brazil). Socioeconomic inequality in self-rated health was measured using the concentration index, which was decomposed to analyze the contribution of different factors. Results: This study revealed that 11.5% of the older adults interviewed reported their health as poor and very poor. For the complete sample, the estimated concentration index, -0.2434, indicated that there is a concentration of poor and very poor self-rated health among older and poorer adults. Income, education and having a private health insurance plan are the factors that contributed most to the observed inequality. Discussion: The decomposition showed that there are avoidable inequalities in relation to socioeconomic status for older adults in Brazil. These factors can guide the formulation of social and health policies aimed at reducing health inequalities.

New highly antigenic linear B cell epitope peptides from PvAMA-1 as potential vaccine candidates.

Peptide-based vaccines have demonstrated to be an important way to induce long-lived immune responses and, therefore, a promising strategy in the rational of vaccine development. As to malaria, among the classic vaccine targets, the Apical membrane antigen (AMA-1) was proven to have important B cell epitopes that can induce specific immune response and, hence, became key players for a vaccine approach. The peptides selection was carried out using a bioinformatic approach based on Hidden Markov Models profiles of known antigens and propensity scale methods based on hydrophilicity and secondary structure prediction. The antigenicity of the selected B-cell peptides was assessed by multiple serological assays using sera from acute P.vivax infected subjects. The synthetic peptides were recognized by 45.5%, 48.7% and 32.2% of infected subjects for peptides I, II and III respectively. Moreover, when synthetized together (tripeptide), the reactivity increases up to 62%, which is comparable to the reactivity found against the whole protein PvAMA-1 (57%). Furthermore, IgG reactivity against the tripeptide after depletion was reduced by 42%, indicating that these epitopes may be responsible for a considerable part of the protein immunogenicity. These results represent an excellent perspective regarding future chimeric vaccine constructions that may come to contemplate several targets with the potential to generate the robust and protective immune response that a vivax malaria vaccine needs to succeed.

Comparative genomics of Leishmania isolates from Brazil confirms the presence of Leishmania major in the Americas.

Leishmania (Leishmania) major is an important agent of cutaneous leishmaniasis, having as a vector sandflies belonging to the genus Phlebotomus. Although this species has been described as restricted to the Old World, parasites similar to L. major have been isolated from South American patients who have never travelled abroad. These parasites were named "L. major-like", and several studies have been carried out to characterise them biochemically, molecularly, and biologically. However, the phylogenetic origin of these isolates is still unknown. In the present study we characterised three L. major-like isolates, named BH49, BH121 and BH129, using comparative genomics approaches. We evaluated the presence of gene and segmental duplications/deletions and the presence of aneuploidies that could explain the differences in infectivity observed in the BH49 and BH121 isolates. All isolates presented a pattern of mosaic aneuploidy and gene copy number variation, which are common in the genus Leishmania. Virulence factors such as phosphatases and peptidases were found to have increased gene copy numbers in the infective isolate, which could explain the difference in infectivity previously observed between BH121 and BH49. Phylogenetic analyses revealed that BH49, BH121 and BH129 L. major-like grouped with L. major isolates, and suggest they were imported from the Old World in at least two independent events. We suggest that new epidemiological inquiries should also evaluate L. major infections in South America, to assess the epidemiological importance of this species in the New World.