Concerns over the replicability and reproducibility of published research have grown in many research fields, but empirical data to inform policies are still scarce. Biomedical research in Brazil expanded rapidly over the last three decades, with no systematic assessment of the replicability of its findings. With this in mind, we set up the Brazilian Reproducibility Initiative, a multicenter replication of published experiments from Brazilian science using three common experimental methods: the MTT assay, the reverse transcription polymerase chain reaction (RT-PCR) and the elevated plus maze (EPM). A total of 56 laboratories performed 143 replications of 56 experiments; of these, 90 replications of 45 experiments were considered valid by an independent committee. Replication rates for these experiments varied between 20 and 44% according to five predefined criteria. In median terms, ratios between group means were 58% lower in replications than in original experiments, while coefficients of variation were 82% higher. Effect size decrease was smaller for MTT experiments, original results with less variability and those considered more challenging to replicate, while t values for replications were positively correlated with researcher predictions about replicability, and negatively correlated with the rate of publications by the original article’s last author. Deviations from preregistered protocols were very common in replications, most frequently due to reasons inherent to the experimental model or related to infrastructure and logistics. Our results highlight factors that limit the replicability of results published by researchers in Brazil and suggest ways by which this scenario can be improved.
The Mayaro virus (MAYV) is an arbovirus that circulates mainly in tropical forests or rural areas in Latin America and is transmitted mainly by Haemagogus mosquitoes. The objective of this study was to evaluate the vector competence, microbiome, and the presence of Wolbachia in three Aedes albopictus populations infected with MAYV. The vector competence was assessed based on viral infection and transmission by RT-qPCR. In addition, the microbiome was evaluated by amplification of the 16S rRNA V4 region and PCR to detect the presence of Wolbachia (strain wAlbA/wAlbB). Our results show that all three populations were susceptible to MAYV infection. The potential transmission of the MAYV was consistent in all populations of naïve mosquitoes injected (more than 50%). The microbiome analysis revealed 118 OTUs (operational taxonomic unit) from the three populations, 8 phyla, 15 classes, 26 orders, 35 families, 65 genera, and 53 species. All populations had Pseudomonas and Wolbachia as predominant genera. There was no difference between the variables for MAYV and Wolbachia (wAlbA or wAlbB) in the abdomen. However, in the head + thorax samples at 14 dpi, there was a difference between the two populations, indicating a possible correlation between the presence of Wolbachia (wAlbB) and infection. Overall, we show evidence that Ae. albopictus displays significant infection and transmission competence for the MAYV in the laboratory, and its bacterial microbiota play an important role in the host, mainly the strains of Wolbachia. The influence of the intestinal microbiota of Ae. albopictus is poorly known, and a better understanding of these interactions would open new perspectives for disease control through the manipulation of microbial communities. The exact contribution of this mosquito species to the transmission of the MAYV in the field remains to be confirmed.
Cell membranes and the cytoskeleton play crucial roles in the regulation of cellular responses by mediating mechanical forces and physical stimuli from the microenvironment through their viscoelastic properties. Investigating these properties provides valuable insights into disease mechanisms and therapeutic strategies. Gold nanorods (GNRs), especially under irradiation, exhibit lethal effects against Leishmania parasites through plasmonic photothermal conversion. In this study, we focus on evaluating the effects of non-irradiated GNRs on macrophage properties to better understand their intrinsic interactions with cells and support the development of future phototherapy applications. Here, defocusing microscopy (DM), a quantitative phase microscopy technique, was used to analyze membrane fluctuations in macrophages () exposed to GNRs (average length of and diameter of ) and infected with Leishmania amazonensis. By quantifying membrane–cytoskeleton fluctuation from defocused images, we extracted viscoelastic parameters, including bending modulus () and viscosity (), to characterize membrane behavior in detail. Our results show that infection increases both and , while treatment at IC50 reduces infection and selectively increases without affecting . In healthy macrophages, exposure to GNRs resulted in a reduction in both parameters, indicative of increased membrane fluidity and cytoskeletal rearrangement. These findings provide new insights into the biomechanical effects of GNRs on macrophages and may enlighten the design of future phototherapeutic approaches.
Cell membranes and cytoskeleton play a crucial role in regulating cellular responses by mediating mechanical forces and physical stimuli from the microenvironment through their viscoelastic properties. Investigating these properties provides valuable insights into disease mechanisms and therapeutic strategies. Gold nanorods (GNRs), especially under irradiation, exhibit lethal effects against Leishmania parasites through plasmonic photothermal conversion. However, their mechanical interaction with cells remains poorly understood. Here, Defocusing Microscopy (DM), a quantitative phase microscopy technique, was employed to analyze membrane fluctuations in macrophages (M∅s) exposed to GNRs and infected with Leishmania amazonensis. Measuring the membrane-cytoskeleton fluctuation of defocused images, we quantified viscoelastic parameters such as bending modulus (kc) and viscosity (η), enabling a detailed char-acterization of membrane behavior. Results indicate that infection increases both kc and η, while IC50 treatment reduces infection and selectively increases kc without affecting viscosity. In healthy macrophages, GNRs reduced both parameters, suggesting a fluidizing effect. These findings contribute to a deeper understanding of GNRs' therapeutic performance and cytotoxicity, contributing to improved phototherapy applications.