Biology subjects

Pinto, A.

Publications and source records attributed to Pinto, A..

At least 19 recordsLinked to original sources

bioRxiv2026

Select properties of supramammillary input to adult-born dentate gyrus neurons

The hypothalamic supramammillary nucleus (SUM) projects to the hippocampal dentate gyrus (DG), a brain area crucial for learning, memory, and adult neurogenesis. How SUM inputs regulate adult-born DG (abDG) neurons to support memory processing remains unclear. Here, using monosynaptic tracing in adult mice, we identified a distinct subpopulation of abDG-targeting SUM neurons defined by large somata and low firing rate. Electrophysiological recordings revealed that these SUM neurons co-release glutamate and GABA onto abDGs, and that voluntary running selectively enhances GABAergic transmission. Chemogenetic silencing of SUM-DG GABAergic projections impaired pattern separation, the ability to make fine distinctions between similar stimuli, but not fear conditioning and spatial navigation memory. Altogether, our findings reveal that a distinct SUM subpopulation provides inhibitory modulation to abDG neurons, establishing a novel circuit mechanism for sparse encoding during pattern separation.

neuroscience↗

bioRxiv2026

Mitophagy is required for cell survival upon ultrasound and microbubble therapeutic exposure

Ultrasound-stimulated microbubble (USMB) sonoporation treatment is an emerging strategy for cancer therapy that enhances drug delivery by perforating the plasma membrane. This membrane permeabilization also causes cellular metabolic stress as cells leak metabolites such as ATP and amino acids. Collectively, cells may die in response to excess membrane damage and stress. However, many cells survive the USMB insult by engaging membrane repair and adaptation pathways such as stimulation of AMPK. In this study, we investigated the impact of USMB treatment on mitochondria and their role in post-exposure cellular survival. We found that USMB causes mitochondrial fragmentation and decreases mitochondrial membrane potential. Subsequently, autophagy and Parkin-dependent mitophagy are triggered post-USMB treatment in an AMPK-dependent mechanism. Importantly, Parkin silencing and PINK1 inhibition augment cell death when combined with USMB exposure, suggesting that mitophagy is an important cell survival process in response to USMB. We propose that combining USMB with inhibitors of mitophagy and metabolic stress adaptation is a strong candidate for therapeutic development to treat solid tumours.

cell biology↗

bioRxiv2026

Capturing domain-level detail of molecular interactions in the IMEx Consortium binding domain data set

The machinery of the cell depends on physical interactions, between proteins themselves, and between proteins and other biomolecules. The detailed characterisation of these interactions, including the identification of the precise sequence regions mediating binding events, is essential to understand both normal cell biology and disease mechanisms. Protein binding domains are specific regions of a protein sequence that mediate direct contact with a partner and represent a critical layer of functional annotation, bridging sequence, structure, and cellular function. Here we present the IMEx binding domain dataset, a large-scale, manually curated open-access set of molecular interactions assembled over more than two decades by the International Molecular Exchange (IMEx) Consortium. The dataset compiles experimentally derived binding region annotations from peer-reviewed literature, systematically linked to protein-protein interaction (PPI) records and mapped to standardised protein sequences in UniProtKB. We describe the curation model, data format, and content of the dataset, provide a descriptive and comparative analysis, and demonstrate the utility of the dataset as a gold standard for benchmarking interface prediction and variant effect annotation methods. Its integration with genomic variations further offers a route to elucidate the mechanistic basis of disease-associated mutations. The datasets are freely available through IntAct in the dedicated PSI-MI FeatureTAB format at https://ftp.ebi.ac.uk/pub/databases/intact/current/psimitab/features/ and are updated and enhanced with every release.

bioinformatics↗

bioRxiv2026

Genome-resolved and kinetic evidence for low-DO comammox-anammox synergy and acetate-stimulated nitrate reduction in IFAS biofilms

Mainstream anammox implementation for nitrogen removal is constrained by unstable nitrite supply and organic carbon requirements for nitrate byproduct removal. This study evaluated integrated fixed-film activated sludge (IFAS) biofilms to enhance anammox activity by coupling low-oxygen ammonium oxidation with volatile fatty acid (VFA)-driven nitrate reduction. Nanopore long-read metagenomic assembly recovered a high-quality, circular single-contig Candidatus Brocadia sapporoensis metagenome-assembled genome (MAG) from full-scale IFAS biofilms. This MAG encodes complete anammox metabolism, dissimilatory nitrate reduction to ammonium (DNRA) and acetate/propionate carbon transformation pathways. Metatranscriptomics showed that low dissolved oxygen (DO) upregulated Ca. B. sapporoensis genes involved in anammox, nitrate reduction, and carbon metabolism. Microaerobic assays established a DO level of 0.7 mg/L as optimal for sustaining near-maximal ammonium oxidation alongside anammox-driven total inorganic nitrogen (TIN) loss. Anoxic tests conducted in secondary effluent indicated that external acetate amendment promoted greater partial nitrate reduction and TIN loss than additional propionate amendment. Integrating this dissolved oxygen concentration with external acetate amendment in a two-stage microaerobic-anoxic system successfully achieved sequential ammonium oxidation, partial nitrate reduction, and anammox-mediated TIN removal. Stage-specific expression suggested Ca. B. sapporoensis could contribute to nitrite self-supplementation via nxrAB-mediated nitrate reduction. Overall, microaerobic ammonium oxidation and Ca. B. sapporoensis-driven partial nitrate reduction jointly sustain mainstream anammox activity. Furthermore, this study demonstrates that successful metabolic synergy depends fundamentally upon precise dissolved oxygen control and effective external acetate amendment.

microbiology↗

bioRxiv2026

Plant actin networks are rapid mechano-adaptive scaffolds

The actin cytoskeleton is among the most highly conserved eukaryotic structures. In animal cells, actin plays a key role in mechanobiology, serving as a structural scaffold that shapes cellular mechanics and a mechanoresponsive network that reorganizes in response to mechanical cues. In plants, these roles are generally attributed to the cell wall and the microtubule cytoskeleton. This leaves the role of plant actin in mechanical responses largely unclear. Here, we ask whether a key actin feature that is essential for animal mechanobiology may also be conserved in plants. We use quantitative live-cell imaging and quantitative image analysis to show that plant actin networks are rapidly mechano-responsive. We find a quantitative correlation between the cellular distribution of cortical actin networks and cellular deformations induced by laser ablation. This process is rapid and completes within 2-4 minutes. We propose that this is an intrinsic physical property of crosslinked filamentous networks, as these findings can be reproduced in a physical model that lacks biological regulation. Moreover, actin networks also respond to different types of mechanical stresses: both osmotic treatment and squeezing lead to distinct changes in actin organization, distribution, and dynamics. Our findings show that plant actin networks form mechano-adaptive scaffolds that sense mechanical deformations and reorganize in response within just a few minutes. This gives rise to a picture of plants having a two-geared cytoskeletal mechano-response system in which microtubules mediate slow responses to subtle developmental stresses, whereas actin provides a rapid response to large and potentially damaging deformations.

plant biology↗

bioRxiv2026

Volumetric Flow Imaging Microscopy to Enhance Particle Characterization

Flow imaging microscopy (FIM) is an important technology for high-throughput characterization of microscopic particles and microorganisms. However, conventional FIM relies on single-plane imaging (SPI), resulting in out-of-focus particles, reduced measurement precision, and incomplete characterization of irregularly shaped objects extending along the z-axis. To address these limitations, a volumetric flow imaging (VFI) framework was developed and implemented on the portable ARTiMiS platform. This approach captures multiple frames along the z-axis and extracts the highest fidelity image for each particle, which can also be used for single image generation with all particles in focus (i.e., all in focus image) and for three-dimensional reconstruction of irregularly shaped objects. Benchmarking VFI with microspheres, live cells (Chlorella vulgaris), and filamentous cyanobacteria demonstrated increased fraction of particles in focus, reduced variability in particle size measurement, and increased resolvability of elongated particles in comparison to conventional SPI on commercially available FIM technologies. For C. vulgaris, VFI-derived size distributions closely matched curated FlowCam measurements without requiring post-processing to exclude out-of-focus particles. All-in-focus image reconstruction enabled simultaneous visualization of particles distributed across multiple depths and consistently resolved a greater proportion of filamentous structures as compared to SPI. For Aphanizomenon sp., Dolichospermum sp., and Planktothrix agardhii, the SPI approach captured only 84%, 61%, and 58%, respectively, of the total filament length resolved by AIF reconstruction. Beyond image-based characterization, VFI enabled estimation of dynamic particle properties such as sinking velocity and mass density. Application of this framework to C. vulgaris cultures revealed distinct mass-density trajectories under nitrogen-replete and nitrogen-deplete conditions, with cell mass density increasing over time under nitrogen-replete conditions and decreasing under nitrogen deprivation. Collectively, these results establish VFI as a next-generation framework for FIM that expands its analytical capabilities beyond conventional morphometric characterization and provides new opportunities for single-cell-enabled environmental monitoring and biomanufacturing.

Cell Biology↗

bioRxiv2026

Inbreeding depression by polygenic load following a severe population bottleneck

It is poorly understood how populations survive extreme bottlenecks despite severe inbreeding. We investigate the genomic architecture of inbreeding in the once critically endangered Seychelles warbler (Acrocephalus sechellensis) using 37 years of individual-based monitoring and 1.8 million SNPs. Linkage disequilibrium patterns reveal a historical effective population size of [~]270 that plummeted to [~]13 coinciding with human colonisation events of the Seychelles archipelago. Contemporary genomes are over one-third inbred from recent inbreeding measured by runs of homozygosity (ROH) formed since human colonisation of the Seychelles, 50 generations ago (mean FROH < 50 generations ago = 0.38), and we identify significant inbreeding depression across cross key fitness traits. On average, a 17.9% reduction in lifespan and a 15.1% reduction in lifetime offspring production were associated with each 10% increase in individual inbreeding. Genome-wide scans reveal this depression is caused by a polygenic load. Our results suggest that a history of small population size could have facilitated the selective purging of severe genetic load, while mildly deleterious alleles escaped selection via drift. A partitioned genetic load architecture potentially enabled species recovery despite the inbreeding depression that followed near-extinction.

evolutionary biology↗

bioRxiv2026

Rap1 Activates Endosomal AC9 to Drive the Second cAMP Wave

GsPCR signaling orchestrates cAMP production in three distinct spatial waves originating from the plasma membrane (PM), endosomes, and the nucleus. While the molecular drivers of the PM and nuclear waves are well defined, the regulation of the endosomal cAMP wave remains insufficiently understood. Here, we identify the small GTPase Rap1b as a direct activator of the endosomal adenylyl cyclase 9 (AC9), revealing a novel mechanism for intracellular cAMP synthesis. Like Gs, Rap1b-GTP interacts with the C2 domain of AC9, allosterically enhancing its catalytic activity both in vitro and in cells. Using AC9 mutations that selectively disrupt Rap1b versus Gs binding, we elucidate the role of the Rap1b-AC9 unit in mediating the endosomal cAMP wave, introducing a new layer of spatial regulation to cAMP signaling.

cell biology↗

bioRxiv2026

A Seychelles warbler genomic toolkit

Understanding evolutionary processes is greatly facilitated by high-quality data on genetic variation. We report the development of a genomic toolkit for a recently bottlenecked, long-term studied species, the Seychelles warbler (Ptimerl dezil; Acrocephalus sechellensis). This toolkit comprises a reference genome assembled into 31 chromosomes, together with functional annotations and reference-panel-free imputation of whole-genome sequences from 1,935 individuals. The genomic data have been used to assign the sequenced individuals into a genetic pedigree. Individual genomic data are associated with a suite of phenotypic metadata, amassed from three decades of fieldwork in this closed, long-term monitored population. We compared sex and parentage assigned using the genomic data with the previously recorded sex and parentage metadata to identify and correct 41 sample DNA samples labelled with the wrong identity. This population resource enables a wide range of analyses, that include, but are not limited to phylogenetics, metabarcoding, recombination rates, linkage patterns, adaptation, heritability, demographic history, selection, and inbreeding estimates. We wish to encourage interest from researchers seeking to collaborate on future analyses and data collection. Overall, our methods demonstrate the potential of next generation sequencing and statistical tools to provide dense genomic datasets at large sample sizes for wild populations.

genomics↗

bioRxiv2026

Scaling Multiplex qPCR Primer Design to 1000-plex using the Degenerate Incomplete Multiplex Primer List Extension (DIMPLE) Algorithm

Massively multiplexed qPCR is primarily constrained by increasing primer dimer formation as the number of distinct primers in a single reaction increases. Previous multiplex primer design algorithms either fail to sufficiently suppress primer dimers at 100+ plex, or take exceedingly high amounts of computational resources to complete. Here, we present DIMPLE, a linear-runtime primer design algorithm that effectively generates 10,000+ primers to amplify thousands of potential amplicons in a single qPCR reaction. As one clinical demonstration of this algorithm, we designed an assay to detect 2,302 distinct KMT2A gene fusion subtypes using 204 primers in a single tube. In contrast to FISH and convention NGS approaches with 2% variant allele frequency (VAF) limit of detection, our DIMPLE qPCR assay was able to analytically detect gene fusions down to 0.05% VAF. We also constructed proof-of-concept multiplex qPCR panels for additional oncology gene fusions, multiplex pathogen detection, and DNA methylation markers. The scalability and low computational cost DIMPLE are complementary to new instrument platforms for massively multiplex qPCR readout for enabling rapid, point-of-care nucleic acid testing.

bioengineering↗

bioRxiv2026

Legionella and Mycobacterium populations exhibit geographic structuring across and within drinking water systems

Opportunistic pathogens (OPs) within the Legionella and Mycobacterium can persist and sometimes proliferate in drinking water systems and pose a risk to public health. Most prior research has focused on isolated system components of the drinking water treatment and distribution system and has rarely examined spatiotemporal dynamics across the entire source water, treatment process, and distribution system continuum. This study addresses this critical knowledge gap by quantitative profiling of microbial communities with full length 16S rRNA gene sequencing and flow cytometry, and associated water chemistry parameters, including disinfection byproducts (DBPs), across five full-scale utilities. These utilities reflect varying source water types, geographic locations, treatment regimes, and climate zones. Microbial communities, including Legionella and Mycobacterium populations, in distribution system were shaped by source water type and exhibited significant community divergence across utilities. Within the same genus, strain-level analyses revealed highly distinct Legionella and Mycobacterium sequence variants unique to each utility. Interestingly, a substantial proportion of Legionella and Mycobacterium amplicon sequence variants were both utility specific and often specific to locations within the distribution system, indicating strong geographic structuring both across and within drinking water systems. Understanding the mechanistic underpinnings of this geographic structuring is critical to develop robust strategies for managing and monitoring Legionella and Mycobacterium populations in drinking water systems.

microbiology↗

bioRxiv2025

MAPK Signaling and Angiopoietin-2 Contribute to Endothelial Permeability in Capillary Malformations

Capillary malformations (CM) are slow-flow vascular abnormalities present at birth and predominantly manifest as cutaneous lesions. In the rare neurocutaneous disorder known as Sturge Weber Syndrome (SWS), individuals exhibit CM not only on the skin but also within the leptomeninges of the brain and the choroid of the eye. >90% of CM are caused by a somatic R183Q mutation in GNAQ, the gene encoding Gq - a heterotrimeric G-protein subunit. The somatic GNAQ mutation is notably enriched in endothelial cells (ECs) isolated from CM-affected regions. Here we show blood vessels in cutaneous and leptomeningeal SWS lesions exhibit extravascular fibrin indicating a compromised endothelial barrier. Longitudinal MRI of the brain in one SWS patient further suggests vascular permeability. To explore this pathological phenotype, we employed the trans-endothelial electrical resistance (TEER) assay to measure permeability of the EC-EC barrier in vitro. Human EC CRISPR edited to create a GNAQ R183Q allele (EC-R183Q) exhibited a reduced barrier compared to mock edited EC (EC-WT). We sought to identify signaling molecules needed for EC barrier formation. Knockdown of angiopoietin-2 (ANGPT2), known to be significantly increased in EC-R183Q and in CM, partially yet significantly restored the barrier, while an anti-ANGPT2 function blocking antibody did not. We next tested the MEK1,2 inhibitor (Trametinib) because MAPK signaling is increased by GNAQ mutation. MEK1,2 inhibitors partially restored the EC barrier, implicating involvement of MAPK/ERK signaling. The combination of ANGPT2 knockdown and Trametinib significantly restored the EC barrier to near EC-WT levels. The additive impacts of ANGPT knockdown and MEK1,2 inhibition indicate the two operate in separate pathways. In summary, we discovered that GNAQ p.R183Q ECs exhibit compromised endothelial barrier formation, reflecting the compromised EC barrier in CM lesions, and that ANGPT2 knockdown combined with Trametinib effectively restores the EC-EC barrier. NONSTANDARD ABBREVIATIONS AND ACRONYMS O_TBL View this table: org.highwire.dtl.DTLVardef@e1081eorg.highwire.dtl.DTLVardef@1dc05b7org.highwire.dtl.DTLVardef@1fb6ac0org.highwire.dtl.DTLVardef@11d8f59org.highwire.dtl.DTLVardef@1ae5406_HPS_FORMAT_FIGEXP M_TBL C_TBL NOVELTY AND SIGNIFICANCEO_ST_ABSWhat is known?C_ST_ABSO_LIThe mutant Gq-R183Q in endothelial cells activates phospholipase {beta}3, contributing to increased angiopoietin-2, a pro-angiogenic, proinflammatory molecule that contributes to vascular permeability. C_LIO_LIEndothelial Gq-R183Q is sufficient to drive formation of enlarged blood vessels akin to what is observed in CM. ANGPT2 shRNA knockdown prevented the enlarged vessel phenotype in a xenograft model. C_LIO_LIAn EC-specific GNAQ p.R183Q mouse model showed permeability in brain vessels, detected by perfusion of Evans Blue dye, indicating reduced vascular integrity. C_LI What New Information Does This Article Contribute?O_LIReduced vascular integrity in CM is confirmed by Martius Scarlet Blue staining and longitudinal MRI imaging of SWS brain. C_LIO_LIGNAQ p.R183Q EC form a weaker endothelial barrier in vitro compared to control ECs. The weakened endothelial barrier in the mutant ECscan be rescued by Gq inhibitor, YM254890, confirming the compromised barrier is a consequence of the mutant Gq. C_LIO_LITitration experiments modeling the mosaic nature of the GNAQ p.R183Q in CMshow that 5- 10% GNAQ p.R183Q EC in the monolayer is sufficient to reduce endothelial barrier formation. C_LIO_LIKnockdown of ANGPT2 or MEK1,2 inhibition partially restored the endothelial barrier in GNAQ p.R183Q EC. C_LIO_LICombining knockdown of ANGPT2 and addition of a MEK inhibitor, Trametinib, restored the endothelial barrier to near what is seen in wild type ECs. C_LI What is the translational message?Sturge Weber Syndrome (SWS) is a neurocutaneous disorder that involves atypical blood vessel overgrowth in the skin, brain and eye. It is associated with facial CM (aka port wine birthmark), leptomeningeal CM in the brain visible with MRI, and glaucoma. Theneurological sequalae involve seizures, cerebral atrophies and calcification, and intellectual disorders. Currently there are no molecularly targeted therapies for non-syndromic CM or SWS. Our study shows the involvement of MAPK pathway and the proinflammatory molecule ANGPT2 in endothelial permeability and suggests a path to target GNAQ p.R183Q driven CM.

pathology↗

bioRxiv2025

CRISPR-Cas9 Screening Reveals Microproteins Regulating Adipocyte Proliferation and Lipid Metabolism

Small open reading frames (smORFs) encode microproteins that play crucial roles in various biological processes, yet their functions in adipocyte biology remain largely unexplored. In a previous study, we identified thousands of smORFs in white and brown adipocytes derived from the stromal vascular fraction (SVF) of mice using ribosome profiling (Ribo-Seq). Here, we expand on this work by identifying additional smORFs related to adipocytes using the in vitro 3T3-L1 preadipocyte model. To systematically investigate the functional relevance of these smORFs, we designed a custom CRISPR/Cas9 guide RNA (sgRNA) library and screened for smORFs influencing adipocyte proliferation and differentiation. Through a dropout screen and fluorescence-assisted cell sorting (FACS) of lipid droplets, we identified dozens of smORFs that regulate either cell proliferation or lipid accumulation. Among these, we validated a novel microprotein as a key regulator of adipocyte differentiation. These findings highlight the potential of CRISPR/Cas9-based screening to uncover functional smORFs and provide a framework for further exploration of microproteins in adipocyte biology and metabolic regulation. SignificanceObesity and its associated metabolic disorders pose significant public health challenges, yet the molecular mechanisms regulating adipocyte function remain incompletely understood. Small open reading frames (smORFs) and their encoded microproteins represent an emerging class of regulatory elements with potential roles in metabolism. Here, we leveraged CRISPR/Cas9 screening to functionally characterize smORFs in adipocytes, identifying novel regulators of cell proliferation and lipid metabolism. Our findings demonstrate that conservation is not a prerequisite for smORF function, as we validated a mouse-specific microprotein that modulates adipocyte differentiation. This work establishes a robust pipeline for unbiased smORF discovery and highlights the potential for species-specific microproteins to regulate adipose biology. Future studies in human adipocytes may uncover additional microproteins with therapeutic relevance for obesity and metabolic disease.

cell biology↗

bioRxiv2025

Aging: an inevitable road toward gut microbiota pathoadaptation

Laboratory-raised mice live approximately seven times longer and healthier lives compared to their wild counterparts, due to a standardized healthy diet and limited exposure to environmental stressors1. Aging is associated with increased inflammation and microbial dysbiosis2-4. Collectively, these influence microbiota evolution and may contribute to the enrichment in pathobiont frequency observed in old age4. Alternatively, this increase could stem from a decline in colonization resistance5,6, creating favorable conditions for pathobiont invasion. Here, we sought to test whether aging in healthy, controlled conditions, could prevent the selection of age-associated pathobionts. We have followed the adaptive evolution of a commensal strain of Escherichia coli in the guts of mice of advanced age and found that it acquired several mutations common to bacteria colonizing young mice, which were absent in old animals. This, together with the increase in Akkermansia muciniphila in mice of advanced age, suggest healthy aging7,8. However, mutations acquired exclusively in the older were mainly pathoadaptive, tuning the metabolism to oxygen and iron availability, hypermotility, and biofilm formation. In summary, while the evolutionary signature in the guts of very old mice shows youth-like features that may be associated with longevity, the selection of pathoadaptive traits is magnified in very old age. While suggesting that a breach in colonization resistance is not needed to justify the abundance of age-associated pathobionts, our findings raise the question whether specialized bacteria, as opposing to generalists such as E. coli, will display the same ability to evolve pathoadaptive traits. HighlightsO_LIGut commensals face increasingly personalized selective pressures in the aging gut C_LIO_LIEven healthy aging selects for pathoadaptive traits in the gut microbiota C_LIO_LIE. coli s adaptive pattern better reflects the metabolome than microbiota composition C_LI In BriefPathobionts are often enriched in the microbiota of the elderly. Melo-Miranda et al. showed that irrespectively of limiting the opportunity for gut invasion, the strength of selection for pathoadaptation increases with aging. Yet, the gut environment of extreme ages seems to converge, highlighting the discontinuity of the aging process.

evolutionary biology↗

bioRxiv2025

Muscle Cathepsin B treatment improves behavioral and neurogenic deficits in a mouse model of Alzheimer's Disease

Muscle secretes factors during exercise that enhance cognition. Myokine Cathepsin B (Ctsb) is linked to memory function, but its role in neurodegenerative disease is unclear. Here we show that AAV-vector-mediated Ctsb overexpression in skeletal muscle in an Alzheimers Disease (AD) mouse model (APP/PS1), improves motor coordination, memory function and adult hippocampal neurogenesis, while plaque pathology and neuroinflammation remain unchanged. Additionally, in AD mice, Ctsb treatment modifies hippocampal, muscle and plasma proteomic profiles to resemble that of wildtype controls. Conversely, in wildtype mice, Ctsb expression causes memory deficits and results in protein profiles across tissues that are comparable to AD control mice. In AD mice, Ctsb treatment increases the abundance of hippocampal proteins involved in mRNA metabolism and protein synthesis, including those relevant to adult hippocampal neurogenesis and memory function. Furthermore, Ctsb treatment enhances plasma metabolic and mitochondrial processes, and reduces inflammatory responses. In muscle, Ctsb expression elevates protein translation in AD mice, whereas in wildtype mice mitochondrial proteins decrease. Overall, the biological changes in the treatment groups are consistent with effects on memory function. Thus, skeletal muscle Ctsb application has potential as an AD therapeutic intervention.

neuroscience↗

bioRxiv2025

External and Internal Load Profiles of Male and Female Participants during a Walking Football Practice

BACKGROUNDHigh levels of sedentary behavior among older adults highlight the importance of walking football (WF) in promoting physical activity for healthy aging. This study examines the external and internal load profiles of male and female participants during a WF tournament, addressing a gap in research on game demands and induced load. METHODSThe study involved 176 players aged 50+ participating in a 40-min, 5v5 WF tournament with unlimited substitutions. External load (total and categorized distances) was measured using Global Positioning System (GPS), while heart rate (HR) monitors assessed internal load, including absolute HR and intensity zones based on %HRmax. RESULTSThe proportion of male participants (n=123; 70.3%) was higher than females (n=52; 29.7%), p<.001. They were similar in age (61.6{+/-}8.6 and 60.8{+/-}6.9, respectively). Males covered a higher distance per minute than females, with sex showing a moderate effect (63.3{+/-}10.7 m/min vs. 54.7{+/-}15.8 m/min; p < .001; Cohens dunbiased = 0.69 [0.36; 1.03]), especially in fast walking (41.7{+/-}12.2 m/min vs. 32.6{+/-}16.7 m/min; p < .001; Cohens dunbiased = 0.66 [0.33; 1.00]). Males played more time than females (22:26{+/-}09:47 min:ss vs. 15:41{+/-}07:46 min:ss; p<.001), with moderate effect (Cohens dunbiased = 0.73 [0.40; 1.06]). However, no differences between sexes were identified in the internal load variables, such that the female average %HRmax was 80{+/-}11% and the male was 82{+/-}8% during the practice. CONCLUSIONSOverall, while males generally exhibit higher external loads in WF, both sexes experience similar internal load demands, highlighting WFs potential as a scalable, health-promoting intervention for aging populations.

physiology↗

bioRxiv2024

DDO-adjuvanted influenza A virus nucleoprotein mRNA vaccine induces robust humoral and cellular type 1 immune responses and protects mice from challenge

A challenge in viral vaccine development is to produce vaccines that generate both neutralizing antibodies to prevent infection and cytotoxic CD8+ T-cells that target conserved viral proteins and can eliminate infected cells to control virus spread. mRNA vaccines offer an opportunity to design vaccines based on conserved CD8-targeting epitopes, but achieving robust antigen-specific CD8+ T-cells remains a challenge. Here we tested the viral-derived oligonucleotide DDO268 as an adjuvant in the context of a model influenza A virus (IAV) nucleoprotein (NP) mRNA vaccine in C57BL/6 mice. DDO268 safely induced local type I interferon (IFN) production, stimulated dendritic cells type 1 (DC1) activation and migration to the draining lymph nodes, and improved the generation of IgG2c antibodies and antigen-specific effector Th1 CD4+ and CD8+ T-cells (IFN{gamma}+TNF+IL2+) when co-packaged with NP mRNA. The DDO268 adjuvanted vaccine provided enhanced protection against lethal IAV challenge and reduced the antigen dose required to achieve this protection. These results highlight the potential of DDO268 as an effective mRNA vaccine adjuvant and show that an IAV NP mRNA/DDO268 vaccine is a promising approach for generating protective immunity against conserved IAV epitopes. IMPORTANCEVaccines that generate neutralizing antibodies and cytotoxic CD8+ T-cells targeting conserved epitopes are ideal for effective protection against viruses. mRNA vaccines combined with the right adjuvant offer a promising solution to this challenge. We show that the virus-derived oligonucleotide DDO268 enhances antibody and T cell responses to an influenza A virus (IAV) nucleoprotein (NP) mRNA vaccine in mice. DDO268 safely induces local type I interferon production and stimulates dendritic cell activation providing enhanced protection against IAV challenge. In addition, the adjuvant activity of DDO268 allows for the use of lower antigen doses during vaccination.

immunology↗

bioRxiv2024

Building plumbing influences the microdiversity and community assembly of the drinking water microbiome.

Building plumbing microbial communities can significantly influence water quality at the point of use, particularly during periods of stagnation. Thus, a fine-scale understanding of factors governing community membership and structure, as well as environmental and ecological factors shaping building plumbing microbial communities is critical. In this study, we utilized full-length 16S ribosomal RNA (rRNA) gene sequencing to investigate the microdiversity and spatial-temporal dynamics of microbial communities in commercial and residential building plumbing systems. Bacterial operational taxonomic units (OTUs) within commercial buildings exhibited much lower microdiversity relative to the same OTUs in residential buildings. Microdiversity was associated with higher persistence and relative abundance of OTUs. Interestingly, amplicon sequencing variants within the same OTUs exhibited habitat preferences based on the building type while also demonstrating varying temporal turnover patterns. Dispersal limitation disproportionately governed community assembly in commercial buildings, whereas heterogeneous selection was the dominant ecological mechanism shaping the microbial community in residential buildings. Dispersal limitation in commercial buildings is consistent with larger building sizes and greater periods of water stagnation. Interestingly, the inability to explain the extent of heterogeneous selection-driven community assembly in residential locations using measured water chemistry may suggest a disproportionately large effect of fine-scale variation in plumbing characteristics on community assembly in residential locations.

microbiology↗