Fundación Instituto Leloir

Brucella infection is frequently acquired by inhalation, but the pathogen disseminates systemically from the lungs. However, little is known about the interaction of Brucella spp. with the airways. Using a 3D air-exposed organotypic human bronchial tissue model (polarized 16HBE14o- bronchial epithelial cells grown over a collagen matrix containing MRC-5 lung fibroblasts), we analyzed Brucella abortus replication, translocation and cytokine responses over prolonged post-infection times. Apically inoculated B. abortus invaded, replicated and persisted during the whole follow-up (16 days) within the bronchial tissue without inducing cytotoxicity. Viable bacteria were also detected in the conditioned medium (CM) since day five post-infection, indicating release from the basolateral side. In parallel experiments, no invasion or bacterial release was detected for Escherichia coli. The levels of IL-6, IL-8 and MCP-1 were increased in CM from Brucella-infected 3D cultures and in monocultures of polarized bronchial epithelial cells or lung fibroblasts. Collagenase/gelatinase activity was increased in 3D cultures and MRC-5 monocultures. Infection transference from bronchial cells to lung fibroblasts was documented using monocultures. An immune cross-talk was detected, as cytokine levels were increased in fibroblasts stimulated with bronchial CM, and vice versa. These results suggest that the bronchial mucosa can sustain B. abortus persistence, replication and dissemination, and that it induces a proinflammatory response to which both epithelial cells and fibroblasts contribute.

Circadian clocks orchestrate daily physiology and behavioral rhythms, yet the extent to which cells lacking canonical clock components exhibit robust temporal regulation remains unclear. Here we addressed this question using a two-step strategy. First, we systematically mapped core clock gene expression at single-cell resolution across the Drosophila brain and body. Second, leveraging these data and performing targeted experiments, we uncovered cells that lack most or all canonical clock components yet display strong mRNA rhythms. We found that lamina wide-field neurons show high-amplitude tim mRNA cycling even in constant darkness conditions (DD) despite minimal expression of other clock genes, suggesting a partial or noncanonical oscillator. In addition, C2 and C3 optic lobe neurons, which do not express core clock components, exhibit hundreds of circadian and daily cycling transcripts. Notably, circadian rhythms in C3 neurons coincide with oscillations of activity-regulated genes (ARGs), whereas C2 neurons cycle independently of ARGs, indicating distinct non-cell-autonomous mechanisms. These findings reveal a spectrum of circadian regulation, from autonomous to remote, input-driven rhythms and expand the circadian landscape to include strategies for generating and distributing temporal information across neuronal and non-neuronal cell types.

Including soybean hulls (SH) in ruminant diets increase the carbon circularity of livestock systems. We evaluated whether SH could partially replace corn grain (CG) in the diet of lactating cows (≈7.5% of the total diet). Six Holstein multiparous cows were fed a 50:50 (dry matter basis) corn-silage to concentrate diet for two 25-day periods in three replicated 2 × 2 Latin Squares using linear mixed models. Feed intake, milk yield and composition, N excretion, CH4 production, and ruminal microbial profiles were measured. Enteric CH4 was measured in respiration chambers. The DMI of cows fed SH (28 kg DM/d) was greater (p = 0.034) than those fed CG (26 kg DM/d), but no differences were observed in milk production (p = 0.557) or enteric CH4 emission intensity (g/kg milk, p = 0.387). Enteric CH4 emissions tended to be greater in SH-fed cows (484 g/d) than CG-fed cows (456 g/d; p = 0.075), and N excretion was significantly greater in SH than CG cows (p < 0.001). No mayor shifts in ruminal microbiota or overall ruminal environment were observed. In conclusion, SH can partially replace CG in diets of lactating dairy cows without affecting production, but potential negative environmental impacts should be considered.

Summary Liquid–liquid phase separation (LLPS) is a key process underlying the formation of biomolecular condensates (BMCs), such as membrane-less organelles (MLOs), that compartmentalize biochemical processes inside the cells. While LLPS has been extensively studied in eukaryotes, its role in bacteria, archaea, and viruses remains far less characterized. Recent studies in bacteria have revealed that LLPS-driven condensates play critical roles in RNA processing, stress response, and pathogenicity. Similarly, many viruses exploit LLPS to facilitate crucial steps in their infection cycles, including viral entry, genome replication, assembly, and host immune evasion. In this work, we introduce a hand-curated database of LLPS proteins from bacteria, archaea, and viruses (BAV-LLPS Database). This resource, extended through sequence similarity searches, comprises over 5,000 proteins and integrates diverse data including biological annotations, sequence features, predicted disordered regions, LLPS per site probability, and AlphaFold2-based structural models. Additionally, our web server enables users to explore both the curated and homologous derived datasets, providing a platform to uncover evolutionary relationships and intrinsic and differential properties of LLPS proteins across various taxonomic groups. This work seeks to deepen our understanding of LLPS mechanisms beyond eukaryotic organisms, emphasizing their significance across diverse life forms. It also aims to foster the development of specialized predictive tools that will facilitate the exploration and characterization of LLPS processes in a wide array of living organisms, thereby contributing to advancements in both fundamental biological research and applied biomedical sciences. Availability and Implementation BAV-LLPS DB is freely accessible at https://bav-llps-db.bioinformatica.org/ . The data can be retrieved from the website. The source code of the database can be downloaded from https://bav-llps-db.bioinformatica.org/download Contact gusparisi@gmail.com and gboccaccio@leloir.org.ar

Epilepsy affects over 50 million individuals worldwide. Despite the availability of anti-seizure medications, approximately 30% of patients remain drug-resistant, emphasizing the pressing need for alternative therapeutic strategies. In recent years, NMR-based metabolomics has emerged as a robust platform to investigate metabolic disturbances in neurological disorders, potentially improving diagnostic precision and explore individualized treatment options.In this study, we analysed serum metabolomics profiles from 32 patients with epilepsy, evaluated both at baseline and shortly after seizure episodes, and 28 healthy controls. Using an untargeted NMR-based metabolomics approach combined with multivariate and statistical analyses, we identified significant metabolic alterations and assessed their diagnostic potential. A total of 14 metabolites differed significantly between patients and controls. Among these, citrate, glutamate, proline, 3-methyl-2-oxovalerate, and glucose showed strong potential as biomarkers. Post-seizure samples revealed alterations in seven metabolites, including hippurate, pyroglutamate, isovalerate, creatinine, threonine, 3-methyl-2-oxovalerate, and 2-oxoisocaproate. Notably, we observed distinct metabolic signatures distinguishing focal from generalized seizures.To our knowledge, this is the first comprehensive serum metabolomics study using NMR to evaluate both basal and post-seizure states in epilepsy. Our findings highlight key alterations in energy metabolism, oxidative stress, and amino acid pathways, offering promising leads for improving clinical assessment and tailoring therapeutic strategies in epilepsy care.Supplementary InformationThe online version contains supplementary material available at 10.1038/s41598-025-14718-1.

Reactive oxygen species (ROS), predominantly derived from mitochondrial respiratory complexes, have emerged as key molecules influencing cell fate decisions like maintenance and differentiation. These redox-dependent events are mainly considered to be cell intrinsic in nature, on the contrary our observations indicate involvement of these oxygen-derived entities as intercellular communicating agents. In Drosophila male germline, Germline Stem Cells (GSCs) and neighbouring Cyst Stem Cells (CySCs) maintain differential redox thresholds where CySC have higher redox-state compared to the adjacent GSCs. Disruption of the redox equilibrium between the two adjoining stem cell populations by depleting Superoxide Dismutases (SODs) especially Sod1 results in deregulated niche architecture and loss of GSCs, which was mainly attributed to loss of contact-based receptions and uncontrolled CySC proliferation due to ROS-mediated activation of self-renewing signals. Our observations hint towards the crucial role of differential redox states where CySCs containing higher ROS function not only as a source of their own maintenance cues but also serve as non-autonomous redox moderators of GSCs. Our findings underscore the complexity of niche homeostasis and predicate the importance of intercellular redox communication in understanding stem cell microenvironments.