Biology subjects

Moreno, A.

Publications and source records attributed to Moreno, A..

At least 19 recordsLinked to original sources

bioRxiv2026

Detection of merbecoviruses in Danish bats using a novel broad-range RT-qPCR assay

Bat-associated merbecoviruses comprise a genetically diverse group of coronaviruses with variable receptor usage and host range, highlighting the need for efficient tools for their surveillance. Here, we developed and optimized a broad-range reverse transcription quantitative PCR (RT-qPCR) assay for the detection of bat-associated merbecoviruses, including the lineage previously identified in brown long-eared bats (Plecotus auritus) in Denmark. In silico analysis predicted detection of 898 of 941 (95.4%) merbecovirus sequences, and the assay detected all bat-associated merbecoviruses included in the experimental panel. The assay was applied to 33 bats sampled through the Danish national bat surveillance program in 2025. Merbecovirus RNA was detected in two samples, one from a P. auritus and one from a Nathusius' pipistrelle (Pipistrellus nathusii), representing the first detection of a merbecovirus in P. nathusii in Denmark. Metagenomic next-generation sequencing recovered a complete merbecovirus genome from the P. nathusii sample. Phylogenetic analysis assigned the virus to the MOW clade and showed that it was closely related to the Russian MOW15-22 virus, sharing 98.67% nucleotide identity across the complete genome and 99.85% amino acid identity in the spike protein. Limited sequence data obtained from the P. auritus sample was consistent with the merbecovirus lineage previously identified in this species in Denmark. Together, these findings demonstrate the utility of broad-range RT-qPCR combined with metagenomic sequencing for surveillance and genomic characterization of bat-associated merbecoviruses.

microbiology↗

bioRxiv2026

Genetic diversity and phylogeography of swine influenza A virus in 2013-2022 in Europe using a harmonized genotyping nomenclature

Swine Influenza A virus (swIAV) poses constant health threat for both humans and animals. This study comprehensively analysed the genetic diversity and evolution of European swIAVs from 2013 to 2022. To address existing classification gaps, an easily accessible full-genome genotyping tool based on Nextclade was developed alongside a global nomenclature system. Application of this tool to European sequence data revealed unprecedented genetic diversity, successfully identifying 150 swIAV genotypes mixing segments of both swine and human origin. Bayesian phylogeographic models, combined with economic data, identified that the trade of live weaner pigs for fattening drove the transboundary spread of swIAVs across Europe. The study detected repeated human-to-swine reverse-zoonotic events across multiple European countries. These findings clearly highlight the role of commercial pig movements in viral dissemination and emphasize the critical importance of harmonized genomic surveillance to monitor swIAV viral evolution to inform global pre-pandemic preparedness strategies.

microbiology↗

bioRxiv2026

Widespread SARS-CoV-2 infection in free-ranging Neotropical bats suggests repeated human-to-bat spillback

Bats harbor exceptional coronavirus diversity and are considered ancestral sources of several human pathogens. As SARS-CoV-2 transitioned from pandemic emergence to global endemicity in humans, concern has shifted from wildlife-to-human spillover toward reverse zoonosis. However, infection of free-ranging bat populations under natural conditions has not previously been demonstrated. Here, we report widespread detection of SARS-CoV-2 RNA in wild Neotropical bats sampled across Andean and Amazonian ecosystems of Southern Ecuador. RT-qPCR screening of 126 individuals, representing nine taxa, detected SARS-CoV-2 RNA in 34.12% of bats across multiple sites. Partial to near-complete viral genomes recovered from five individuals showed >99% nucleotide identity to contemporary human SARS-CoV-2 lineages and clustered within multiple global phylogenetic clades. Mixed-effects modeling revealed pronounced species-level heterogeneity, a positive association between elevation and infection probability, and higher infection probability in females compared with males. The close phylogenetic affinity of bat-derived genomes to circulating human variants and their distribution across multiple lineages suggest repeated anthropogenic spillback rather than sustained bat-specific circulation. These results expand current understanding of the ecological footprint of the COVID-19 pandemic and highlight the importance of integrating wildlife surveillance into long-term One Health strategies for emerging infectious diseases.

microbiology↗

bioRxiv2026

A spatially-specific pattern of cortical thickness in frontal lobe predisposes to faster future amyloid accumulation in individuals with subjective cognitive decline

The risk of developing AD is conferred through numerous genetic and environmental factors. How these risks influence brain structure prior to the onset of AD pathology is not yet known. We hypothesise that there may be specific anatomic endophenotypes that predispose to AD years before the first signs of amyloid accumulation occur. Using clinical, genetic and brain MRI data from Fundacio ACE Healthy Brain Initiative (FACEHBI) --a longitudinal cohort of individuals with Subjective Cognitive Decline aged > 50 years--, we identified a spatially-specific, fine-grained pattern of cortical thickness in frontal lobe regions among amyloid-negative individuals at baseline that holds a significant relationship with the rate of amyloid accumulation during the next five years. This pattern may represent a structural fingerprint of increased susceptibility to the initiation of the AD cascade.

neuroscience↗

bioRxiv2026

Immunological imprinting shapes the cross-reactive antibody responses to the KP.2 and LP.8.1 vaccine doses

The emergence of the SARS-CoV-2 Omicron BA.2.86 subvariant, a lineage derived from the BA.2 strain, led to the 2024-2025 COVID-19 vaccine update to include KP.2 or related JN.1-lineage spike antigens. We evaluated the magnitude, breadth, and durability of humoral immune responses following a single KP.2 vaccine dose in a longitudinal cohort of 21 individuals up to six months. KP.2 vaccination increased spike-specific binding and neutralizing antibodies against the ancestral WA1 strain, alongside the BA.5, XBB.1.5, and KP.2 variants. Power law modeling estimated half-lives for WA1- and KP.2-specific IgG responses at 770 and 248 days, respectively. Additionally, the KP.2 dose increased IgG1 and IgG4 subclasses more than IgG2 and IgG3 responses to both spike proteins. Serum depletion experiments using WA1 or KP.2 proteins demonstrated most vaccine-elicited antibodies were cross-reactive. Consequently, KP.2 vaccine-induced antibodies retained broad neutralizing activity against recently circulating Omicron subvariants (BA.2.86, KP.3.1.1, XEC, LP.8.1, LF.7, XFG.3.12, PQ.1, BA.3.2.1, and RE.2). Using a live virus neutralization assay, XFG.3.12 showed the greatest reduction in neutralizing titers relative to KP.2 (4.2-fold). In a small subset, an LP.8.1 vaccine dose increased neutralizing activity against the matched variant while maintaining WA1 and KP.2 cross-reactivity, but only modestly increased antibodies to divergent variants BA.3.2.1 and RE.2. Ultimately, these data indicate the KP.2 mRNA vaccine generates durable, cross-reactive responses against current Omicron subvariants. However, ongoing spike evolution impacts neutralization of emerging lineages, highlighting the need for continued viral monitoring and timely vaccine updates. IMPORTANCESARS-CoV-2 continues to evolve, raising ongoing concerns about how well updated vaccines protect against emerging variants. This study evaluates antibody responses after KP.2 spike mRNA vaccine dose and shows that a single dose induces durable and broadly cross-reactive immunity against both earlier strains and recently circulating Omicron subvariants. Despite this breadth, reduced neutralizing activity against certain emerging variants indicates that ongoing antigenic changes can impact vaccine induced antibody effectiveness. These findings provide insight into how current vaccines perform over time and highlight the need to track viral evolution and update vaccine antigens to maintain broad protection against severe disease, hospitalization, and death.

immunology↗

bioRxiv2026

Dirty mice better recapitulate key features of mRNA vaccine immunogenicity observed in humans

Although specific pathogen free (SPF) mice have traditionally been used to test candidate vaccines, recent work has demonstrated that "dirty" mice with broad microbial exposure more appropriately recapitulate human immune responses. Using a model where lab mice are co-housed with pet store mice, we modeled SARS-CoV-2 mRNA vaccine responses in dirty and traditional SPF models. We found that dirty mice show reduced serum spike-binding antibody titers after prime and require a second booster dose to reach SPF-level spike antibody titers. Additionally, spike antibodies showed faster waning in dirty mice through 5 months post-vaccination, and neutralizing activity of these antibodies were reduced against Omicron variants, directly comparable with observations in humans. We further investigated the seasonality and consistency of pathogens in cohoused mice, and the impact of serial microbial exposure on our animal model system. We found that pathogen exposure and T cell activation remained consistent over time, and that a single co-housing event was sufficient to provide broad microbial exposure. This work demonstrates that the dirty mouse co-housing system is a promising, translationally representative approach to screen candidate mRNA vaccines for efficacy and durability prior to human clinical trials. Significance statementThe development of mRNA vaccines during the COVID-19 pandemic dramatically reduced hospitalization and death rates for infected individuals. However, booster vaccinations were required to achieve full efficacy, and protection waned over time. Our research leveraged a "dirty" mouse model to test whether SARS-CoV-2 mRNA vaccinations in animals with previous microbial exposure better modelled human immune responses. We found that dirty mice require a booster vaccination for full efficacy and experienced waning serum antibody titer over time. We propose this approach as a future model for robust preclinical mRNA vaccine testing.

immunology↗

bioRxiv2025

Languages of the brain: fMRI dissection of the amodal networks for language, mathematics, and social knowledge

The ability to compose complex mental representations by recombining simpler primitives is a characteristic of the human brain that manifests itself in a variety of domains such as spoken or written language, mathematics, or social reasoning. While it is known that these domains rest on partially distinct brain networks, they are rarely investigated together in the same paradigm. Here, we present a systematic functional MRI study of written and spoken word and sentence processing in each of these domains. In a true-false judgment task, adult participants were presented with a hierarchy of meaningless and meaningful stimuli bearing on various domains of general semantic knowledge and mathematics. The results show that distinct brain regions are activated by (1) syntax and semantics; (2) math versus non-math knowledge; (3) different domains of mathematics, such as geometry versus arithmetic; (4) social knowledge versus other knowledge. All of these regions are activated essentially identically in the written and spoken modalities. The results were replicated in a group of adolescents. Our experiments show that the human brain comprises distinct amodal networks for various domains of linguistic and semantic knowledge, and provide a simple paradigm to dissect them within a short fMRI session.

neuroscience↗

bioRxiv2025

CryoEM of ATP-driven dynamics and itraconazole binding in a fungal ABC pump

Azole resistance in Candida species is often caused by the overexpression of Cdr1. Despite its clinical relevance, the structural basis for its ATP-driven efflux pump function remains elusive. We present four high-resolution cryo-EM structures for Candida glabrata Cdr1 under active turnover conditions in the absence and presence of ATP-Mg{superscript 2}, itraconazole, and vanadate. Additional transient cryo-EM structures were unveiled by 3D variability analysis offering a detailed view of the step-by-step transitions triggered by ATP-hydrolysis. The motion cascade starts with a 4 [A] piston-like retraction of the C-helix from the {gamma}-phosphate/vanadate of the hydrolyzed ATP. This causes the nearby transmembrane helix-1 (TMH-1) to open the drug-binding site via lateral displacement and unwinding of the inner-leaflet region of TMH-2. A reverse squeeze-and-push motion of TMH-2 possibly drives substrate extrusion. High resolution structures also reveal how itraconazole adapts its shape to fit into the drug-binding site. Our findings provide a dynamic structural framework for Cdr1-mediated azole resistance and the conserved chemo-mechanical cycle of ABC proteins, including non-membranous members.

biochemistry↗

bioRxiv2025

Aminopeptidase N is a receptor for hedgehog merbecoviruses

Merbecoviruses, closely related to the highly pathogenetic Middle East Respiratory Syndrome Coronavirus (MERS-CoV), circulate in hedgehogs throughout Europe and Asia, raising concerns about zoonotic transmission to humans and domestic animals. Unfortunately, how these viruses enter host cells remains unknown, hindering experimental studies. Here, we tested known coronavirus receptor orthologues from European hedgehogs (Erinaceus europaeus) and identified Aminopeptidase N (APN) as an entry receptor for hedgehog merbecoviruses. We confirm this result with single-cycle pseudotype and replication-competent virus experiments as well as protein binding assays. A screen of 30 mammalian APN orthologues reveals restricted cross-species receptor use. Cryo-electron microscopy analysis of the viral glycoprotein-receptor complex shows a unique interface distinct from known coronavirus spike:APN interactions, providing a molecular basis for species barriers. These findings expand the known range of receptor use not only within merbecoviruses but also betacoronaviruses, improving our understanding of betacoronavirus receptors, and informing risk assessments for viral emergence.

microbiology↗

bioRxiv2025

Systematic molecular glue drug discovery with a high-throughput effector protein remodeling platform

Realising the promise of new medicines that operate through a targeted molecular glue-induced degradation mechanism requires systematic tools that can uncover the relevant principles of neomorphic protein-protein interactions. Whilst some monovalent glue degraders have been found through serendipity, the rules for small molecule attributes and the pairs or complexes of proteins that are amenable to drug-induced proximity control remain poorly articulated. Here we introduce a new approach to address this by using programmed libraries of intramolecularly edited proteins to expand protein surface landscapes and trigger new druggable interactions. We show that effector proteins, such as the E3 ligase Cereblon, can be engineered to provoke neomorphic activity by inducing the degradation of new client proteins and that these de novo interactions provide a blueprint from which new small molecule degraders can be built. As a demonstration of the approach, we use the platform to identify new non-IMiD molecular glue degraders of the oncology target GSPT1. SUMMARYO_LIMolecular glues are a highly important and promising new form of therapeutic agent, but rationalising their discovery has so far been impossible C_LIO_LIGlueSEEKER screening enables prospective monovalent drug discovery by using high-throughput deep mutational scanning to re-engineer the function of effector proteins like E3 ligases C_LIO_LIWe used this approach to enable the computational discovery of small molecule glues which degrade the oncology target GSPT1 and show how the technology can be used across new targets C_LI

bioengineering↗

bioRxiv2025

Inactivation of Influenza A Viruses (H1N1, H5N1) During Grana-Type Raw Milk Cheesemaking: Implications for Foodborne Transmission Risk

BackgroundThe detection of H5N1 highly pathogenic avian influenza virus (HPAIV) in lactating dairy cattle in the United States, with high viral titers in raw milk, has raised concerns about potential zoonotic transmission through the consumption of unpasteurized milk and raw-milk dairy products. While inactivation studies exist for pasteurized milk, data on virus persistence during the manufacture of raw-milk cheeses remain scarce. AimTo evaluate the survival and inactivation of avian influenza viruses (AIV), including both low pathogenic (LPAIV, H1N1) and highly pathogenic (HPAIV, H5N1) strains, during the production and ripening of Grana-type hard cheeses made from raw cows milk. MethodsExperimental cheesemaking was conducted using raw milk artificially contaminated with A/duck/Italy/281904-2/06 (H1N1; 107.75 EID50/mL) or A/duck/Italy/326224-2/22 (H5N1 clade 2.3.4.4b; 106.75 EID50/mL). Cheeses were produced in accordance with Parmigiano Reggiano production standards and ripened for 30 days at 5-6 {degrees}C. Viral presence was assessed in finished cheeses by inoculation on SPF embryonated chicken eggs (ECE), hemagglutination (HA) assay, and monoclonal antibody-based ELISA. ResultsNo infectious virus was detected in any cheese sample produced from contaminated milk following two blind passages in SPF-ECE. Both HA and ELISA tests yielded negative results, indicating complete inactivation of the virus. ConclusionThis study demonstrates that the traditional Grana-type cheese production process-- including curd cooking, acidification, and ripening--effectively inactivates both LPAIV and HPAIV, even at high contamination levels. These findings support the microbiological safety of hard cheeses made from raw milk with regard to AIV, contributing to risk assessment and food safety policies during avian influenza outbreaks.

microbiology↗

bioRxiv2025

Regulation of B cell tolerance and autoimmunity by miR-130b

Breakdown of B cell tolerance is a central feature of autoimmune diseases, yet the molecular mechanisms underlying the female predominance of these diseases remain unclear. Here, we identified miR-130b as an essential component of a miRNA network, defined by the GUGCA seed motif, that regulates central B cell tolerance. Elevated miR-130b levels impaired tolerance by promoting the survival of immature B cells upon B cell antigen receptor engagement. This network converged on the estrogen receptor pathway through downregulation of Esr1 and Pten. Genetic ablation of Esr1 in immature B cells was sufficient to compromise central B cell tolerance in females, but not in males, revealing an unrecognized role for ER in this process. In males, Esr1 deficiency reduced bone marrow B cell numbers, whereas in females, B cell numbers were preserved, with a greater proportion of cells displaying lower surface CD19 levels. Transcriptomic analysis revealed sex-specific genetic programs characterized by defective B cell development and skewed immunoglobulin gene usage in males, and increased PI3K-AKT signaling in females. In females, this molecular rewiring compensated for developmental defects, but also attenuated central tolerance mechanisms, allowing the escape of autoreactive B cells to the periphery. In patients with multiple sclerosis, elevated levels of miR-130b in circulating vesicles correlated with more severe disease, including increased formation of new demyelinating lesions, cognitive decline, and neurodegeneration. Together, our findings identify a sex-specific ER-dependent checkpoint in B cell tolerance controlled by a seed-driven miRNA network, providing a mechanistic framework for female predisposition to autoimmunity.

immunology↗

bioRxiv2024

From North to South: Transmission Dynamics of H1N1pdm09 Swine Influenza A Viruses in Italy

The influenza A H1N1pdm09 virus continues to be a significant pathogen, posing potential risks to both animal and human health due to its zoonotic potential. Italy, which has one of the largest swine populations in Europe, plays a crucial role in monitoring the evolution of influenza viruses in livestock. This study aims to address the existing knowledge gaps regarding the genetic diversity and transmission dynamics of H1N1pdm09 circulating in Italian swine populations. Utilizing whole genome sequencing and dynamic modeling, we conducted a comprehensive analysis of virus samples collected from swine farms across Italy. Our results reveal that multiple independent viral introductions have occurred into the country, with most cases resulting in self-limited infections and limited onward transmission. However, six distinct transmission clusters were identified, suggesting instances of sustained viral spread. These clusters were found across multiple regions of Italy, highlighting the broad geographic distribution of virus lineages. Our findings indicate that while many introductions led to localized containment, certain virus lineages were able to spread within specific regions of Italy. Through a detailed examination of selective pressures, we observed that most viral genes are under strong purifying selection in both swine and human hosts, as reflected by dN/dS ratios well below 1. The hemagglutinin (HA) gene exhibited a notably higher dN/dS ratio in swine ([~]0.28) compared to humans ([~]0.22), indicating slightly relaxed selection in swine. In contrast, other genes, such as neuraminidase (NA) and non-structural protein (NS), showed similarly strong purifying selection across both hosts. These results reflect a general trend of selective pressures affecting multiple viral components, rather than emphasizing specific genes. Our study emphasizes the importance of ongoing genomic surveillance in detecting viral circulation and mitigating risks to both animal and public health. Italys efforts contribute significantly to global influenza monitoring and highlight the importance of a One Health approach that integrates human, animal, and environmental health. These findings provide essential data to inform public health policies and enhance preparedness against future zoonotic influenza outbreaks.

microbiology↗

bioRxiv2024

Modelling the prebiotic origins of regulation & agency in evolving protocell ecologies

How did natural systems develop the first mechanisms of regulation and what for? How could they turn into adaptive agents in a minimal (though deeply meaningful) biological sense? A novel simulation platform, Araudia, has been implemented to address these questions, which are deeply interrelated, in a prebiotic scenario where metabolically diverse protocells are allowed to modify their dynamic behaviour in response to changes in their boundary conditions (e.g., nutrient concentrations in the medium) and/or in the activity of other protocells, including cross-feeding relationships. We extend consumer-resource models (CRMs) to a stochastic evolutionary framework in which novelty appears bottom-up (i.e., from small changes at the individual protocell level), and a short-term memory may also come forth and spread in the population, to demonstrate that simple adaptive/learning processes can have relevant effects at somatic times (i.e., within the lifetime of single protocells). Our interest in exploring the interplay between metabolic-physiological aspects and ecological-evolutionary ones derives from the fact that it provides a complex causal domain, where various spatial and temporal scales intermingle, and where both the actual and the potential (pathways/behaviours) must be considered. It is in such a complex domain where the appearance of regulation acquires full significance, as an effective means to navigate phenotypic spaces that become too big for unguided exploration, given the large number of possible functionalities (or functional states) accessible to each proto-cellular agent. The results obtained for several minimalist cases below show that, despite the non-trivial combination of --both endogenous and exogenous--factors involved in the constitution of regulatory mechanisms, these are bound to play a critical role in the survival of infra-biological systems, especially under rapidly changing environmental conditions that longer-term adaptive mechanisms would not be able to cope with.

ecology↗

bioRxiv2024

Enhancing Gene Set Overrepresentation Analysis with Large Language Models

Gene set overrepresentation analysis is widely used to interpret high-throughput transcriptomics and proteomics data, but traditional methods rely on static, human-curated gene set databases that lack flexibility. We introduce llm2geneset, a framework that leverages large language models (LLMs) to dynamically generate gene set databases tailored to input query genes, such as differentially expressed genes (DEGs) and a biological context specified in natural language. These databases integrate with methods, such as overrepresentation analysis (ORA), to assign biological functions to input genes. Benchmarking against human-curated gene sets demonstrates that LLMs generate gene sets comparable in quality to those curated by humans. llm2geneset also identifies biological processes represented by input gene sets, outperforming traditional ORA and direct LLM prompting. Applying the framework to RNA-seq data from iPSC-derived microglia treated with a TREM2 agonist highlights its potential for flexible, context-aware gene set generation and improved interpretation of high-throughput biological data. llm2geneset is available as open source at https://github.com/Alector-BIO/llm2geneset and via a web interface at https://llm2geneset.streamlit.app.

bioinformatics↗

bioRxiv2024

Semaglutide Administration in Healthy Mice Alters Behaviour Related to Stress and Motivation

Semaglutide, a GLP-1 receptor agonist, is widely studied in metabolic disorders, yet its effects in healthy subjects remain unclear. Here, we show that daily administration of semaglutide (0.1 mg/kg) in healthy mice alters behaviour in various tests related to stress and reward pursuit. Additionally, acute administration of semaglutide changes the oscillatory activity in the nucleus accumbens in the delta, theta and alpha bands. These findings reveal that semaglutide impacts both behaviour and neural dynamics in non-diseased brains, offering a baseline for interpreting its broader therapeutic potential.

neuroscience↗

bioRxiv2024

Persistent legacy effects on soil metagenomes facilitate plant adaptive responses to drought

Both chronic and acute drought alter the composition and physiology of soil microbiota by selecting for functional traits that preserve fitness in dry conditions. Currently, little is known about how the resulting precipitation legacy effects manifest at the molecular and physiological levels and how they influence neighboring plants, especially in the context of subsequent drought. We characterized metagenomes of six prairie soils spanning a steep precipitation gradient in Kansas, USA. By statistically controlling for variation in soil porosity and elemental profiles, we identified bacterial taxa and functional gene categories associated with precipitation. This microbial precipitation legacy persisted through a 5-month-long experimental drought and mitigated the negative physiological effects of acute drought for a wild grass species that is native to the precipitation gradient, but not for the domesticated crop species maize. In particular, microbiota with a low-precipitation legacy altered transcription of a subset of host genes that mediate transpiration and intrinsic water use efficiency during drought. Our results show how long-term exposure to water stress alters soil microbial communities with consequences for the drought responses of neighboring plants.

microbiology↗

bioRxiv2024

Representation of grasping type and force in the primate motor cortex

The primary motor cortex (M1) is strongly engaged by movement planning and movement execution. However, the role of M1 activity in voluntary grasping is still not completely understood. Here we analyze recordings of M1 neurons during the execution of a delayed reach-to-grasp task, where monkeys had to actively grasp an object with either a side or a precision grip, and then pull it with a low or high amount of force. Single cell and neural populations analyses showed that grip type was robustly and specifically encoded by a large population of neurons, while force level was weakly encoded within mixed-selective neurons that also provided grip type information. Notably, the grip type was stably decoded from motor cortical populations during the preparation and execution epochs of the task. Our results are consistent with the idea that planning and performing specific grasping movements are high-level skills that strongly engage M1 neurons, while the execution of grasping-pulling force might be prominently encoded at lower stages of the motor system. Significance statementGrasping behavior requires precise motor coordination exerted by multiple brain areas, including the primary motor cortex (M1), but the exact role of M1 in grasping preparation and execution remains elusive. Here, we analyzed the neural activity of M1 while two monkeys performed a delayed reach-to-grasp task. We found that two parameters of grasping: grip type and force level, were represented in the activity of single neurons and the neural population, although with important differences. While grip representation was stronger and more temporally stable, force encoding was weaker and short lived. Our results suggest that grip planning and execution is a high-level neural process that takes place independently of force control in M1.

neuroscience↗