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Abstract Despite all the efforts acquired in four years of the COVID-19 pandemic, the path to a full understanding of the biological mechanisms involved in this disease remains complex. This is partly due to a combination of factors, including the inherent characteristics of the infection, socio-environmental elements, and the variations observed within both the viral and the human genomes. Thus, this study aimed to investigate the correlation between genetic host factors and the severity of COVID-19. We conducted whole exome sequencing (WES) of 124 patients, categorized into severe and non-severe groups. From the whole exome sequencing (WES) association analysis, four variants (rs1770731 in CRYBG1, rs7221209 in DNAH17, rs3826295 in DGKE, and rs7913626 in CFAP46) were identified as potentially linked to a protective effect against the clinical severity of COVID-19, which may explain the less severe impact of COVID-19 on the Northern Region. Our findings underscore the importance of carrying out more genomic studies in populations living in the Amazon, one of the most diverse from the point of view of the presence of rare and specific alleles. To our knowledge, this is the first WES study of admixed individuals from the Brazilian Amazon to investigate genomic variants associated with the clinical severity of COVID-19.

Abstract Natural lignocellulosic fibers (NLFs) have shown a great potential as reinforcements in composites in recent decades. Among other reasons, environmental concerns and the depletion of oil reserves justify research on natural composites as they offer an environmentally friendly alternative and align with the principles of sustainable development. Among the plethora of NLFs available in nature, the ubim fiber (Geonoma baculifera) has not yet been investigated as a reinforcement for composites in potential engineering applications. Therefore, this study evaluates, for the first time, the mechanical properties of epoxy composites with 10, 20, and 30 vol% of ubim fibers. These properties were assessed through Izod impact, tensile and flexural tests as well as dynamic mechanical analysis (DMA). The data were statistically analyzed using the ANOVA method. Scanning electron microscopy (SEM) analysis indicated a transition from a purely brittle fracture mechanism to a ductile-brittle combination as the fiber volume in the composite increased. Tensile tests of the composites demonstrated an increasing trend in strength and elastic modulus with fiber volume. The results of the flexural tests also displayed a similar trend in strength and elasticity modulus for the composites. The results of DMA tests showed that composite materials with a 30 vol% of ubim fibers exhibited a high glass transition temperature and a low tan δ value, suggesting higher stiffness of this composite compared to others. Overall, the results indicated that the incorporation of 30 vol% ubim fibers into the composites significantly improved their mechanical properties compared to other tested fiber fractions. Additionally, their functional characteristics, such as simplicity in the manufacturing process, low cost, and excellent strength-to-weight ratio, make these composites particularly suitable for applications in sectors such as the automotive industry, construction panels, and packaging. These factors contribute to the development of an efficient, sustainable, recyclable and environmentally friendly composite.

Abstract Plasmodium vivax, a challenging species to eliminate, causes millions of malaria cases globally annually. Developing an effective vaccine is crucial in the fight against vivax malaria, but considering the limited number of studies focusing on the identification and development of P. vivax-specific vaccine candidates, exploring new antigens is an urgent need. The merozoite protein CyRPA is essential for P. falciparum growth and erythrocyte invasion and corresponds to a promising candidate antigen. In P. vivax, a single study with multiple vaccine candidates indicates PvCyRPA with strong association with protection, outperforming classic malaria vaccine candidates. However, little is known about the specific naturally acquired response in the Americas, as well as the antigen epitope mapping. For this reason, we aimed to investigate the cellular and humoral immune response elicited against PvCyRPA in Brazilian endemic areas to identify the existence of immunodominant regions and the potential of this protein as a single or even a multi-stage specific malaria vaccine candidate for P. vivax. The results demonstrated that PvCyRPA is naturally immunogenic in Brazilian Amazon individuals previously exposed to malaria, which presented anti-PvCyRPA cytophilic antibodies. Moreover, our data show that the protein also possesses important immunogenic regions with an overlap of B and T cell epitopes. These data reinforce the possibility of including PvCyRPA in vaccine formulations for P. vivax.

Abstract Measures reflecting cardiac sympathovagal activity, particularly those associated with heart rate variability (HRV), are widely recognized and utilized in both scientific and clinical contexts. This study aimed to assess the inter- and intra-examiner reliability of short-term HRV parameters in patients hospitalized with coronavirus disease 2019 (COVID-19). A total of 103 patients (both sexes) diagnosed with COVID-19 were included in the study. HRV was analyzed using both linear and nonlinear methods. Reliability was evaluated through intraclass correlation coefficient (ICC2.1), minimum detectable change (MDC), standard error of measurement (SEM), and coefficient of variation (CV). According to Fleiss’ criteria, excellent reliability was demonstrated, with ICC values ranging from 0.970 to 0.999 for Examiner 1, and from 0.956 to 0.999, for Examiner 2. In the inter-examiner analysis, the ICCs of HRV parameters ranged from 0.972 to 0.999. SEM values for intra-examiner reliability for Examiner 1 ranged from 0.02 to 5.64, with MDC values from 0.05 to 15.64, and CV (%) from 0.28 to 8.04. For Examiner 2, SEM values ranged from 0.02 to 8.18, MDC values from 0.05 to 22.68, and CV (%) from 0.24 to 8.14. For inter-examiner reliability, SEM values ranged from 0.02 to 6.17, MDC from 0.06 to 17.11, and CV (%) from 0.34 to 9.81. Across all analyses, CVs for HRV parameters remained below 10%. Considering different time points and different examiners, short-term resting HRV measurements in patients hospitalized with COVID-19, as evaluated using a portable heart rate device, exhibit high reliability.

Abstract The ultrastructure of human oocytes has been described only qualitatively. To offer a precise organelle spatial distribution and organelle volume during the main maturation stages, we previously conducted stereological studies on prophase-I (GV) and metaphase-I (MI) oocytes, and here we present results on metaphase-II (MII) oocytes. Five donor oocytes from different donors were processed for transmission electron microscopy, and quantification of organelle distribution was performed using point-counting stereology. Statistical tests compared the means of the relative volumes occupied by organelles among oocyte regions. The most abundant organelles were elements of the smooth endoplasmic reticulum (SER), such as SER small vesicles, SER medium vesicles, SER large vesicles and SER isolated tubules, along with mitochondria, followed by SER tubular aggregates, cortical vesicles and lysosomes. Significant differences between oocyte regions were found for lysosomes, cortical vesicles and SER large vesicles. Comparisons of MII oocytes to previous findings in GV and MI oocytes evidenced specific patterns of organelle distribution and relative volumes. This final evaluation thus enables to track organelle spatial reorganization across oocyte stages, which, in addition to gathered knowledge, may be useful to assist in improvements of stimulation protocols, in-vitro maturation media and cryopreservation techniques.

Abstract Alzheimer’s disease (AD) is a progressive neurodegenerative condition and the primary form of dementia among elderly people. The amyloidogenic hypothesis is the main theory that explains this phenomenon and describes the extracellular accumulation of amyloid beta (Aβ) peptides. Model organisms such as Drosophila melanogaster have been utilized to improve the understanding of this disease and its treatment. This study evaluated the effects of peptide and metabolic fractions of Brazilian kefir on a strain of D. melanogaster that expresses human Aβ peptide 1–42 in the eye. The parameters assessed included ommatidial organization, vacuole area, retinal thickness, and Aβ peptide quantification. The present study revealed that the fractions, particularly the peptidic fraction, significantly reduced the vacuole area and increased the retina thickness in treated flies, indicating an improvement in neurodegeneration phenotype. The peptidic fraction was also found to alter Aβ aggregation dynamics, inhibiting Aβ fibril formation, as revealed by dynamic light scattering. This study demonstrated that kefir fractions, particularly the peptidic fraction

Abstract Although heart rate variability (HRV) is a valid method to evaluate the behavior of the autonomic nervous system in individuals with temporomandibular disorder (TMD), the measurement can easily be biased by factors involving the analysis methodology, such as the removal of artifacts. Therefore, the objective of this investigation is to evaluate the impact of using different levels of threshold-based artifact correction to process HRV data in individuals with TMD. This cross-sectional observational study. Adults aged 18 to 55 years old with a diagnosis of myogenic TMD, score ≥ 50 on the Fonseca Anamnestic Index (FAI) and pain ≥ 3 on the Numerical Pain Scale (NPS) participated. The HRV was registered in the supine position (short-term) using a Polar S810i. Kubios software was used for HRV analysis using all filters. One-way ANOVA with Tukey-Kramer post-hoc was used to test the differences in HRV using the different Kubios Software artifact correction filters. The effect size was calculated based on the Cohen d. The very strong filter was statistically different (p

Abstract Positive expiratory pressure (PEP) is a technique used in respiratory physiotherapy to treat diseases related to the respiratory system through spontaneous breathing. This equipment consists of an oronasal mask coupled to a T connector with a unidirectional valve. Studies that evaluate whether the pressure level in the one-way valve corresponds to the actual pressure level provided are scarce in the scientific literature. In order to investigate the failures, bench tests were carried out on the spring-loaded valves, using a U-tube manometer. This pressure was exerted on the valve using a syringe that generated air flow inside the U-tube, allowing analysis numerical value of the measured pressure and the specified values of the valve, thus verifying the disparity of these measured values in relation to the PEP values operated by valves (0 to 20 cmH2O) from the three manufacturers under study. PEPs generated by spring-loaded valves from all three manufacturers were higher than pressures in the range of 2.5 to 20 cmH2O, with significant differences between manufacturers. This bench study showed inaccurate operation of all spring-loaded PEP valves of the three manufactures. The results obtained and the performance of the valves require a reevaluation of manufacturing procedures to preserve product quality and efficacy in clinical application.

Abstract Coral microbiomes play crucial roles in holobiont homeostasis and adaptation. The host’s ability to populate broad ecological niches and to cope with environmental changes seems to be related to the flexibility of the coral microbiome. By means of high-throughput DNA sequencing we characterized simultaneously both bacterial (16S rRNA) and Symbiodiniaceae (ITS2) communities of four reef-building coral species (Mussismilia braziliensis, Mussismilia harttii, Montastraea cavernosa, and Favia gravida) that differ in geographic distribution and niche specificity. Samples were collected in a marginal reef system (Abrolhos, Brazil) in four sites of contrasting irradiance and turbidity. Biological filters governed by the host are important in shaping corals’ microbiome structure. More structured associated microbial communities by reef site tend to occur in coral species with broader geographic and depth ranges, especially for Symbiodiniaceae, whereas the endemic and habitat-specialist host, M. braziliensis, has relatively more homogenous bacterial communities with more exclusive members. Our findings lend credence to the hypothesis that higher microbiome flexibility renders corals more adaptable to diverse environments, a trend that should be investigated in more hosts and reef areas.

Abstract Sedentary behavior (SB) is associated with health impairments, while physical activity (PA) has been a protective factor. It is unclear whether SB and PA are associated with occurrence of signs and/or symptoms (SS) during cardiac rehabilitation program (CRP) exercise sessions. The objective was to evaluate the association between SB and PA with occurrence of SS. Was included 48 patients from a CRP program (64.7 ± 10.4 years-old). Daily time and % of time of SB and weekly time in moderate-to-vigorous-intensity [MVPA], % of MVPA time, steps/day, and steps/minute were accelerometer-measured. Patients were followed-up during 24 CRP sessions, for accompaniment of SS. Age, sex, and comorbidities (hypertension, diabetes, dyslipidemia, obesity) were covariates. Log-transformed values of SB, MVPA and steps/day were also analyzed. As results, 43.7% (n = 21) of participants presented occurrence of signs, 62.5% (n = 30) presented occurrence of symptoms, and 81.2% (n = 39) present occurrence of SS. In fully adjusted model, % of time in MVPA (β: −0.449,p = 0.045) and steps/minute (β: −0.244,p = 0.026) were inversely associated with occurrence of symptoms. No association was observed between SB and PA and occurrence of signs. The occurrence of symptoms and SS among CRP participants was directly associated with SB and inversely associated with variables of PA.

Abstract The increase in the construction of mega dams in tropical basins is considered a threat to freshwater fish diversity. Although difficult to detect in conventional monitoring programs, rheophilic species and those reliant on shallow habitats comprise a large proportion of fish diversity in tropical basins and are among the most sensitive species to hydropower impacts. We used Baited Remote Underwater Video (BRUV), an innovative, non-invasive sampling technique, to record the impacts caused by Belo Monte, the third largest hydropower project in the world, on fishes inhabiting fast waters in the Xingu River. BRUV were set in a river stretch of ~ 240 km for 7 years, 2 before and 5 after the Belo Monte operation. We explored the spatial and temporal variation in fish diversity (α, β, and γ) and abundance (MaxN) using generalized additive models. We also investigated the variation of environmental variables and tested how much information we gained by including them in the diversity and abundance models. Belo Monte altered the flow regime, water characteristics, and fishery yield in the Xingu, resulting in changes in the fish community structure. Temporally, we observed sharp declines in α diversity and abundance, far exceeding those from a previous study conducted with more conventional sampling methods (i.e., catch-based) in the region. γ-diversity was also significantly reduced, but we observed a non-expected increase in β diversity over time. The latter may be associated with a reduction in river connectivity and an increase in environmental heterogeneity among river sectors. Unexpected signs of recovery in diversity metrics were observed in the last years of monitoring, which may be associated with the maintenance of flow levels higher than those previously planned. These results showed that BRUV can be a useful and sensitive tool to monitor the impacts of dams and other enterprises on fish fauna from clear-water rivers. Moreover, this study enhances our comprehension of the temporal variations in freshwater fish diversity metrics and discusses the prevalent assumption that a linear continuum in fish-structure damage associated with dam impoundments may exhibit temporal non-linearity.

Abstract This study aims to (1) compare the kinetics of pulmonary oxygen uptake (VO2p), skeletal muscle deoxygenation ([HHb]), and microvascular O2 delivery (QO2mv) between heart failure (HF) patients with reduced ejection fraction (HFrEF) and those with preserved ejection fraction (HFpEF), and (2) explore the correlation between body composition, kinetic parameters, and exercise performance. Twenty-one patients (10 HFpEF and 11 HFrEF) underwent cardiopulmonary exercise testing to assess VO2 kinetics, with near-infrared spectroscopy (NIRS) employed to measure [HHb]. Microvascular O2 delivery (QO2mv) was calculated using the Fick principle. Dual-energy X-ray absorptiometry (DEXA) was performed to evaluate body composition. HFrEF patients exhibited significantly slower VO2 kinetics (time constant [t]: 63 ± 10.8 s vs. 45.4 ± 7.9 s; P

Abstract Inherited retinal dystrophies/degenerations (IRDs) are the leading cause of visual impairment and incurable familial blindness in the Western world. Given the clinical and genetic heterogeneity, establishing a molecular diagnosis is especially relevant. The aim of this study was to perform the first nationwide survey to understand the prevalence and current management of IRDs in Portugal. A response was obtained from 26 healthcare providers (HCP) (76.5% response rate). Only 4 respondents reported not managing IRD patients. Most HCPs (68.1%) reported managing up to 100 patients, while three currently manage between 501 and 1000 patients. Based on the Portuguese population, an estimated IRD prevalence of 0.031%, i.e., about 1 in 3000 individuals, was calculated. In most HCPs (86.3%), most patients are adults, and non-syndromic retinitis pigmentosa is the most frequent diagnosis. Only 4 HCPs currently use the national, web-based IRD registry (IRD-PT). However, all but one respondent expressed interest in participating in such a registry. Genetic testing is available in 54.5%, with 58.3% HCPs reporting solved rates between 61–80%, but 4 to 9 months to get a genetic test result in 83.4% of cases. Based on this survey, the prevalence of biallelic RPE65-associated disease in Portugal is 0.00031%, i.e., approximately 1:300,000 individuals. Data from this study provide vital background information on national differences in the diagnosis and management of IRD patients. Nationwide implementation of the IRD-PT registry should be encouraged and supported to provide population-based reference data and to identify patients eligible for current and future therapies.

Abstract Simarouba, a neotropical genus in the family Simaroubaceae, currently lacks comprehensive genomic data in existing databases. This study aims to fill this gap by providing genomic resources for three Simarouba species, S. amara, S. versicolor, and S. glauca. It also aims to perform comparative molecular evolutionary analyses in relation to other species within the order Sapindales. The analysis of these three Simarouba species revealed the presence of the typical quadripartite structure expected in plastomes. However, some pseudogenization events were identified in the psbC, infA, rpl22, and ycf1 genes. In particular, the CDS of the psbC gene in S. amara was reduced from 1422 bp to 584 bp due to a premature stop codon. Nucleotide diversity data pointed to gene and intergenic regions as promising candidates for species and family discrimination within the group, specifically matK, ycf1, ndhF, rpl32, petA-psbJ, and trnS-trnG. Selection signal analyses showed strong evidence for positive selection on the rpl23 gene. Phylogenetic analyses indicated that S. versicolor and S. glauca have a closer phylogenetic relationship than S. amara. We provide chloroplast genomes of three Simaruba species and use them to elucidate plastome evolution, highlight the presence of pseudogenization, and identify potential DNA barcode regions.

Abstract Advances in omics technologies have enabled the in-depth study of microbial communities and their metabolic profiles from all environments. Here metagenomes were sampled from piranha (Serrasalmus rhombeus) and from river water from the Rio São Benedito (Amazon Basin). Shotgun metagenome sequencing was used to explore diversity and to test whether fish microbiomes are a good proxy for river microbiome studies. The results showed that the fish microbiomes were not significantly different from the river water microbiomes at higher taxonomic ranks. However, at the genus level, fish microbiome alpha diversity decreased, and beta diversity increased. This result repeated for functional gene abundances associated with specific metabolic categories (SEED level 3). A clear delineation between water and fish was seen for beta diversity. The piranha microbiome provides a good and representative subset of its river water microbiome. Variations seen in beta biodiversity were expected and can be explained by temporal variations in the fish microbiome in response to stronger selective forces on its biodiversity. Metagenome assembled genomes construction was better from the fish samples. This study has revealed that the microbiome of a piranha tells us a lot about its river water microbiome and function.

Abstract SARS-CoV-2 caused the pandemic situation experienced since the beginning of 2020, and many countries faced the rapid spread and severe form of the disease. Mechanisms of interaction between the virus and the host were observed during acute phase, but few data are available when related to immunity dynamics in convalescents. We conducted a longitudinal study, with 51 healthy donors and 62 COVID-19 convalescent patients, which these had a 2-month follow-up after symptoms recovery. Venous blood sample was obtained from all participants to measure blood count, subpopulations of monocytes, lymphocytes, natural killer cells and dendritic cells. Serum was used to measure cytokines, chemokines, growth factors, anti-N IgG and anti-S IgG/IgM antibodies. Statistic was performed by Kruskal–Wallis test, and linear regression with days post symptoms and antibody titers. All analysis had confidence interval of 95%. Less than 35% of convalescents were anti-S IgM+, while more than 80% were IgG+ in D30. Anti-N IgG decreased along time, with loss of seroreactivity of 13%. Eosinophil count played a distinct role on both antibodies during all study, and the convalescence was orchestrated by higher neutrophil-to-lymphocyte ratio and IL-15, but initial stages were marked by increase in myeloid DCs, B1 lymphocytes, inflammatory and patrolling monocytes, G-CSF and IL-2. Later convalescence seemed to change to cytotoxicity mediated by T lymphocytes, plasmacytoid DCs, VEGF, IL-9 and CXCL10. Anti-S IgG antibodies showed the longest perseverance and may be a better option for diagnosis. The inflammatory pattern is yet present on initial stage of convalescence, but quickly shifts to a reparative dynamic. Meanwhile eosinophils seem to play a role on anti-N levels in convalescence, although may not be the major causative agent. We must highlight the importance of immunological markers on acute clinical outcomes, but their comprehension to potentialize adaptive system must be explored to improve immunizations and further preventive policies.

Abstract: The TIGIT+FOXP3+Treg subset (TIGIT+Tregs) exerts robust suppressive activity on cellular immunity and predisposes septic individuals to opportunistic infection. We hypothesized that TIGIT+Tregs could play an important role in intensifying the COVID-19 severity and hampering the defense against nosocomial infections during hospitalization. Herein we aimed to verify the association between the levels of the TIGIT+Tregs with the mechanical ventilation requirement, fatal outcome, and bacteremia during hospitalization. TIGIT+Tregs were immunophenotyped by flow cytometry from the peripheral blood of 72 unvaccinated hospitalized COVID-19 patients at admission from May 29th to August 6th, 2020. The patients were stratified during hospitalization according to their mechanical ventilation requirement and fatal outcome. COVID-19 resulted in a high prevalence of the TIGIT+Tregs at admission, which progressively increased in patients with mechanical ventilation needs and fatal outcomes. The prevalence of TIGIT+Tregs positively correlated with poor pulmonary function and higher plasma levels of LDH, HMGB1, FGL2, and TNF. The non-survivors presented higher plasma levels of IL-33, HMGB1, FGL2, IL-10, IL-6, and 5.54 times more bacteremia than survivors. Conclusions: The expansion of the TIGIT+Tregs in COVID-19 patients was associated with inflammation, lung dysfunction, bacteremia, and fatal outcome.