Biology subjects

Qi, F.

Publications and source records attributed to Qi, F..

At least 19 recordsLinked to original sources

bioRxiv2026

Efficient recovery of transgene-free edited tobacco using the transgene self-elimination system via de novo regeneration

Elimination of integrated editing constructs remains a major challenge for generating transgene-free progeny in plant genome editing based on Agrobacterium-mediated regeneration. Here, we adapted the Transgene Killer CRISPR (TKC) concept to tobacco and established the transgene self-elimination system, in which CRISPR/Cas9 is coupled with developmentally regulated BARNASE cassettes. Three pU3TR-derived vectors carrying DLL-, OsRP3- or PHAS-driven Barnase regenerated normally after leaf-disc transformation. The DLL construct pEB138 showed the highest T editing frequency, with 56 of 69 independent T plants (81.16%) carrying mutations at one or both NtDFR target loci. Under a stringent lineage-level screen in which up to 48 T progeny were tested per T line, 36 of 43 pEB138 lineages (83.72%) yielded uniformly marker-negative progeny for the three assayed T-DNA markers, compared with 0 of 7 lineages transformed with the parental pU3TR vector. The platform also supported recovery of dfr mutants with altered flower pigmentation and NtA622-edited lines with reduced nicotine accumulation. Moreover, introduction of DLL-iBarnase into the independent pDC45 editing backbone did not impair shoot regeneration, and representative NtBBL progeny showed markedly reduced nicotine content together with loss of the selectable cassette. These results demonstrate that BARNASE-assisted transgene elimination can be integrated with Agrobacterium-mediated de novo regeneration in a dicot, enabling one-step recovery of transgene-free edited tobacco progeny.

molecular biology↗

bioRxiv2026

Widespread Presence of Catalytic RNAs in All Domains of Life

Very few ribozymes have been found in complex eukaryotes. Here, we developed a novel method, AbRiDGed, for highly accurate and sensitive ab initio self-cleaving ribozyme discovery. Our results indicate the existence of millions of self-cleaving ribozymes in humans and other species. We characterized ~150 novel classes of self-cleaving ribozymes, including one class which exhibits noncanonical biochemical properties and widespread presence in all domains of life and appears to play specific functions in complex organisms. These results revealed that the true complexity of the "ribozyme-ome" is several orders of magnitude higher than previously estimated. We demonstrate that newly identified self-cleaving ribozymes influence organismal fitness by regulating gene expression and enhancing transcriptome complexity in vivo. Moreover, these ribozymes hold promise as foundational tools for biotechnological applications, such as direct RNA detection technologies. Overall, catalytic RNAs are prevalent and diverse, and may represent a hidden regulatory layer in various organisms.

molecular biology↗

bioRxiv2026

Deciphering underground decarboxylase activity towards Nε-modified lysine derivatives in enterobacteria.

Thermal food processing generates diverse compounds interacting with the gut microbiota. Despite their abundance, the microbial turnover of diet-borne N{varepsilon}-modified lysine derivatives remains largely unexplored. We demonstrate that the enterobacterial ornithine decarboxylase SpeC degrades the prevalent advanced glycation end product N{varepsilon}-carboxymethyllysine (CML) to carboxymethylcadaverine via an underground activity ([~]4 molecules/enzyme/min). This promiscuity extends to additional N{varepsilon}-modified lysine derivatives - namely fomylated (FmL), monomethylated (MML) and dimethylated (DML) lysine - yielding previously unknown biogenic amines (mono- and dimethylcadaverine, formylcadaverine). Functionally, SpeC enables Escherichia coli to utilize CML as a sole nitrogen source. In specific strains, this metabolism reinforces pH-stress responses, supporting survival under mild acidic conditions typical for the colon. Furthermore, SpeC orthologs are widespread across human gut genomes, correlating with geography, diet, and disease. Together, these findings suggest a potential diet-microbiome communication axis, linking the intake of modified dietary chemicals to microbial physiology and hypothesized host impacts.

microbiology↗

bioRxiv2026

A specialized ARGONAUTE enables trans-species RNA interference in plant immunity

Trans-species RNA interference (tsRNAi), in which plants produce small RNAs (sRNAs) to silence target genes in pathogens, has emerged as a promising strategy for disease control. However, whether tsRNAi constitutes an endogenous, regulated immune response remains unclear. Here, we show that ARGONAUTE10 (AGO10) plays a critical role in pathogen-induced tsRNAi. Loss of AGO10 in Arabidopsis abolished pathogen gene silencing during infection, leading to hypersusceptibility to oomycete and fungal pathogens. Importantly, AGO10 rapidly responds to pathogen infection through increased protein accumulation and re-location into discrete cytoplasmic condensates, thus promoting the production of trans-species sRNAs at the pathogen infection sites. This immune responsiveness relies on the N terminal intrinsically disordered region (IDR) of AGO10, which is responsible for sensing and responding to immune activation. Specific features in the IDR partitions AGO10 into two deeply diverged subgroups, AGO10a and AGO10b, with the immune responsiveness and defense function evolutionarily conserved in AGO10a but not AGO10b. Together, these findings establish tsRNAi as a bona fide, evolutionarily conserved immune response and position AGO10 as a signal-responsive hub linking pathogen perception to tsRNAi-based defense.

plant biology↗

bioRxiv2025

A high-resolution dimeric structure reveals a critical role of alpha-helix 9 in apoptotic Bax pore assembly in the mitochondrial membrane

Bax functions as a proapoptotic protein perforating the mitochondrial membrane to release mitochondrial proteins and DNA that kill the cell. Structures of full-length Bax monomer in solution and core domain dimers in solution and bound to lipid/detergent bicelles reveal how Bax proteins must be activated, unfold, refold and dimerize prior to forming oligomeric pores in the membrane. In particular, amphipathic core dimer forms part of the pore wall between the nonpolar lipid bilayer and the aqueous conduit. However, atomic resolution structures of other parts of the pore were scarce. Here, we elucidate a structure of Bax C-terminal -helix 9 (9) in lysolipid micelles using NMR. According to this high-resolution structure the 9 regions form an amphipathic helical dimer with an extended nonpolar interface and several uncharged polar residues on the surface. Structure-guided mutagenesis and functional assessment demonstrate that the nonpolar interactions are important for Bax dimerization in and perforation of the mitochondrial membrane. Surprisingly the polar residues are also important because they form bifurcated hydrogen bonds between helical turns to stabilize each helix and thereby the dimer. Molecular dynamics simulations of an oligomer constructed with wall-forming core and transmembrane 9 dimers linked by flexible 6-7-8 bridges in a mitochondrial lipid bilayer generate an atomic resolution model for a stable Bax pore capable to release cytochrome C. Thus, we made an important step toward elucidating molecular mechanisms of apoptotic Bax perforation.

biochemistry↗

bioRxiv2025

Cortical microglia promote noradrenergic signaling to maintain wakefulness

Microglia are essential for maintaining brain homeostasis, including the sleep-wake cycle, but the underlying mechanisms remain unclear. Here, using in vivo two-photon imaging in both head-fixed and freely moving mice with simultaneous electroencephalogram-electromyogram (EEG-EMG) recording, we investigated microglial process dynamics and their interactions with locus coeruleus (LC) noradrenergic (NE) axons in the frontal cortex across sleep-wake states. During sleep or chemogenetic suppression of LC neurons, microglia enhanced their surveillance and increased interactions with NE axons. Correlative electron microscopy at nanometer resolution identified structural contacts between microglial process endings and NE boutons. Mechanistically, microglia-bouton interaction is primed by NE-{beta}2-adrenergic receptor ({beta}2AR) signaling, leading to enhanced Ca{superscript 2} activity in NE axons. These findings suggest that microglial interactions with NE axons promote cortical norepinephrine transmission and maintain stable wakefulness.

neuroscience↗

bioRxiv2025

Chronic stress abnormalize microglial-vascular interaction via AT1R signaling and microglia activation to drive depression-like behaviors

Depression entails a range of pathological processes in the brain, typically characterized by microglia activation, neural dysfunction, and vascular dysregulation. While it is well established that activated microglia inflict damage to neurons through direct and indirect microglia-neuron interactions, their interaction with vasculature and the associated regulation of vascular structure and function in depression remain poorly understood. Here, using a mouse model of depression induced by chronic unpredictable stress (CUS), we find that in stressed mice, activated microglia present reduced motility and decreased contact area with vasculature, which is accompanied by pathological vascular structure and function, marked by reduced vessel density, narrowed vascular diameters, and sluggish blood flow velocity. We observe that vascular structure in the stressed mice is rapidly modified by the vascular-contacted microglia. These vascular-contacted microglia show upregulated angiotensin II type 1 receptor (AT1R) expression, which is not present in the vascular-uncontacted microglia. Furthermore, we demonstrate that AT1R signaling together with microglia activation under chronic stress are critical mediators for the altered microglial-vascular contact situation, vascular dysregulation, as well as the depression-like behaviors. Together, our findings reveal that under CUS, the aberrant microglial-vascular interaction, including the dysregulated microglial vasoregulation arising from AT1R signaling and microglia activation, leads to depression, forging a new link between chronic stress and depression.

neuroscience↗

bioRxiv2025

Satellite microglia gate neuronal excitability by shielding inhibitory synapses in adult brain

Microglia, the primary immune cells of the central nervous system, are known for sculpting excitatory neural circuits via dynamic processes. However, their role in modulating inhibitory synaptic connections in adulthood remains largely unexplored. In this study, we identified an underappreciated microglia subpopulation, satellite microglia with physical soma-soma contact with neurons, that reshape inhibitory circuit connectivity and gate neuronal activity by shielding inhibitory synapses in the adult mouse brain. Using chronic in vivo two-photon imaging in adult mice, we observed in real time that satellite microglia associations displaced perisomatic inhibitory synapses whereas their dissociation permitted inhibitory synaptogenesis. Redistribution of satellite microglia reorganized inhibitory connectivity, altered local neural networks and changed behaviors. These findings establish satellite microglia as key architects of inhibitory circuitry and suggest their broader role in neural plasticity in health and disease.

neuroscience↗

bioRxiv2025

Neutrophil-microglia interaction drives reversible motor dysfunction in neuromyelitis optica model induced by subarachnoid AQP4-IgG

Neutrophils and neutrophil extracellular traps (NETs) contribute to early neuromyelitis optica (NMO) histopathology initiated by IgG targeting astrocytic aquaporin-4 water (AQP4) channels. Yet, the mechanisms recruiting neutrophils and their pathogenic roles in disease progression remain unclear. To investigate molecular-cellular events preceding classical complement cascade activation in a mouse NMO model, we continuously infused, via spinal subarachnoid route, a non-complement-activating monoclonal AQP4-IgG. Parenchymal infiltration of netting neutrophils containing C5a ensued with microglial activation and motor impairment, but no blood-brain barrier leakage. Motor impairment and neuronal dysfunction both reversed when AQP4-IgG infusion stopped. Two-photon microscopy and electron-microscopy-based reconstructions revealed physical interaction of infiltrating neutrophils with microglia. Ablation of either peripheral neutrophils or microglia attenuated the motor deficit, highlighting their synergistic pathogenic roles. Of note, mice lacking complement receptor C5aR1 exhibited reduction in neutrophil infiltration, microglial lysosomal activation, neuronal lipid-droplet burden and motor impairment. Pharmacological inhibition of C5aR1 recapitulated this protection. Immunohistochemical analysis of an NMO patients early spinal cord lesions revealed analogous pathological findings. Our study identifies neutrophil-derived C5a signaling through microglial C5aR1 as a key early driver of reversible motor neuron dysfunction in the precytolytic phase of NMO. One Sentence SummaryNeutrophil-derived C5a coactivates microglia to drive reversible motor paresis initiated by a non-complement-activating aquaporin-4-IgG binding to astrocytes in a mouse model of neuromyelitis optica.

neuroscience↗

bioRxiv2025

Microglial SWELL1 Deficiency Drives Male-Specific Seizure Vulnerability but Paradoxical Neuroprotection through Impaired Phagocytosis

The discovery of genes encoding the volume-regulated anion channel (VRAC) has enabled detailed exploration of its cell type-specific roles in the brain. LRRC8A (SWELL1) is the essential VRAC subunit. We observed seizure-induced, subunit-specific changes in microglial VRAC expression and investigated its function using conditional knockout (cKO) of LRRC8A in microglia. SWELL1 cKO mice exhibited a male-specific increase in kainate-induced seizure severity yet showed paradoxical neuroprotection against seizure-associated neuronal loss. Mechanistically, SWELL1 deletion led to a cell-autonomous reduction in microglial density and decreased release of VRAC-permeable neuroactive metabolites, including taurine, GABA, and glutamate in culture. Additionally, impaired phagocytic kinetics and reduced lysosomal biogenesis contributed to the observed neuroprotection. These findings reveal novel roles for microglial VRAC in regulating seizure outcomes and microglia-neuron interactions.

neuroscience↗

bioRxiv2024

Metabolic Divergence Between Healthy and Conditional Presenilin-1/2 knockout Dementia Model Mice under Chronic Nicotine exposure

Nicotine, a primary bioactive compound in tobacco, influences a wide range of metabolic pathways, with potential implications for neurodegenerative diseases such as Alzheimers disease (AD). This study investigates the metabolic effects of chronic nicotine administration in presenilin-1/2 double knockout (DKO) mice and wild-type (WT) control mice. Eight-month-old DKO and WT mice underwent a three-month oral nicotine administration which did not significantly affect body weight or spontaneous locomotor activity in either DKO or WT mice, as measured by weekly body weight measurement and open field test. The metabolomic analyses were performed using untargeted LC-MS and GC-MS techniques. The results revealed distinct metabolic profiles between DKO and WT mice under nicotine exposure. In DKO mice, nicotine exacerbated deficits in purine and nicotinamide metabolism, further highlighting dysregulation in energy homeostasis. Key findings include reduced adenosine and elevated dopamine levels in DKO serum, alongside increased cotinine concentrations, suggesting altered nicotine metabolism. WT mice showed significant changes in energy and lipid metabolism pathways, though with less pronounced alterations compared to DKO mice. Enrichment analyses identified significant pathway disruptions, particularly in choline metabolism, amino acid metabolism, and central carbon metabolism, with genotype-dependent responses to nicotine. This study highlights the intrinsic metabolic alterations in DKO mice, such as disruptions in purine and nicotinamide metabolism, which contribute to their neurodegenerative pathology and unique responses to nicotine. By revealing the differential metabolic impacts of nicotine on healthy and neurodegenerative states, the findings provide valuable insights into its dual therapeutic and pathological roles. These results may guide future research into the metabolic underpinnings of neurodegeneration and the potential for nicotine as a targeted therapeutic intervention.

neuroscience↗

bioRxiv2024

Lack of Ameliorative Effect of Chronic Oral Nicotine on Olfactory Dysfunction in Young Adult Presenilin-1/2 Double Knockout Mice of a Dementia Model

Olfactory dysfunction is a prevalent and early feature of neurodegenerative diseases including Alzheimers disease while the features and underlying mechanisms remain inadequately understood. This investigation aimed to elucidate the olfactory function characteristics of presenilin-1/2 double knockout (DKO) mice, an established mouse model of dementia. Through a series of behavioral tests on DKO and wildtype (WT) mice at 2 and 6 months of age, we assessed their abilities in odor detection, discrimination, and olfactory memory. The findings revealed a substantial deficit in odor detection in 2-month-old DKO mice compared to WT mice, with a noteworthy age-related deterioration observed from 2 to 6 months. Although DKO mice exhibited normal odor discrimination and olfactory memory capabilities at 2 months, these functions declined with age, falling significantly lower levels than observed in WT mice by the age of 6 months. Subsequently, 2-month-old DKO and WT mice underwent a 4-month chronic nicotine treatment through drinking water. This intervention failed to ameliorate olfactory dysfunctions in DKO mice and exhibited no significant impact on WT mice, despite a discernible trend of a potential negative effect. In summary, this behavioral study elucidates that Presenilin-1/2 double knockout impairs odor detection in young adult mice and accelerates declines of olfactory discrimination and memory. The chronic administration of nicotine did not effectively mitigate olfactory function deficiencies in young adult DKO mice, and the results also highlight age as an important contributing factor.

animal behavior and cognition↗

bioRxiv2024

SARS-CoV-2 vaccination during pregnancy enhances hippocampal neurogenesis and working memory in offspring via IFN-gamma responsive microglia

In the face of severe adverse outcomes of pandemic Coronavirus disease 2019 (COVID-19) infection, vaccines proved potently immunogenic and safe in humans and are today strongly recommended in pregnancy. This study investigates, in offspring mice, the effect of maternal COVID-19 vaccination on postnatal physical development, behavior and neurogenesis. After inoculation with inactivated COVID-19 vaccine (Vero Cell) at gestational day 14.5, antibodies to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were detected in serum of both dams and pups. At one month of age, pups born to vaccinated dams, but not the offspring of non-vaccinated dams, exhibited greater working memory and more neural cell proliferation, neuroblast formation, neuronal stem cell activity and larger numbers of mature neurons within the dentate gyrus (DG). Luminex multiplex assay revealed elevated levels of hippocampal cytokines/chemokines critical to neurogenesis and memory function, namely interferon-{gamma} [IFN-{gamma}] and CX3C motif chemokine ligand 1 [CX3CL1]. Conditional knockout technology implicated microglial IFN-{gamma} receptor 1 (IFN{gamma}R1) and CX3C motif chemokine receptor 1 (CX3CR1) as crucial intercellular participants in the neuronal developmental process, via regulating microglial activation and chemotaxis, respectively. We propose that, rather than posing risk for neurodevelopmental abnormalities, maternal SARS-CoV-2 vaccination transiently enhances hippocampal neurogenesis and working memory in offspring.

animal behavior and cognition↗

bioRxiv2024

Rod-shaped microglia interact with neuronal dendrites to regulate cortical excitability in TDP-43 related neurodegeneration

Motor cortical hyperexcitability is well-documented in the presymptomatic stage of amyotrophic lateral sclerosis (ALS). However, the mechanisms underlying this early dysregulation are not fully understood. Microglia, as the principal immune cells of the central nervous system, have emerged as important players in sensing and regulating neuronal activity. Here we investigated the role of microglia in the motor cortical circuits in a mouse model of TDP-43 neurodegeneration (rNLS8). Utilizing multichannel probe recording and longitudinal in vivo calcium imaging in awake mice, we observed neuronal hyperactivity at the initial stage of disease progression. Spatial and single-cell RNA sequencing revealed that microglia are the primary responders to motor cortical hyperactivity. We further identified a unique subpopulation of microglia, rod-shaped microglia, which are characterized by a distinct morphology and transcriptional profile. Notably, rod-shaped microglia predominantly interact with neuronal dendrites and excitatory synaptic inputs to attenuate motor cortical hyperactivity. The elimination of rod-shaped microglia through TREM2 deficiency increased neuronal hyperactivity, exacerbated motor deficits, and further decreased survival rates of rNLS8 mice. Together, our results suggest that rod-shaped microglia play a neuroprotective role by attenuating cortical hyperexcitability in the mouse model of TDP-43 related neurodegeneration.

neuroscience↗

bioRxiv2024

mRNA Vaccine-Induced SARS-CoV-2 Spike-Specific IFN-γ and IL-2 T-cell Responses are Predictive of Serological Neutralization and are Transiently Enhanced by Pre-Existing Cross-Reactive Immunity

The contributions of SARS-CoV-2-specific T-cells to vaccine efficacy and durability are unclear. We investigated the relationships between mRNA vaccine-induced spike-specific IFN-{gamma} and IL-2 T-cell responses, anti-spike/RBD IgG/IgA antibodies, and live virus neutralizing capacity in long-term-care-home staff doubly vaccinated with BNT162b2 or mRNA-1273. The impacts of pre-existing cross-reactive T-cell immunity to SARS-CoV-2 on cellular and humoral responses to vaccination were additionally assessed. Mathematical modelling of the kinetics of spike-specific IFN-{gamma} and IL-2 T-cell responses over 6-months post-second dose was bifurcated into recipients who exhibited gradual increases (54% and 42%, respectively) with doubling times of 173 days, or decreases (46% and 58%, respectively) with half-lives of 115 days. Differences in kinetics did not correlate with any clinical phenotypes, although increases were proposed to be due to subclinical viral exposures. Serological anti-spike/RBD IgG/IgA antibody levels had otherwise decayed in all participants with half-lives of 55, 53, 76, and 59 days, respectively. Spike-specific T-cell responses induced at 2-6 weeks correlated with live viral neutralization at 6-months post-second dose, especially in hybrid immune individuals. Participants with pre-existing cross-reactive T-cell immunity to SARS-CoV-2 exhibited greater spike-specific T-cell responses, reduced anti-RBD IgA antibody levels, and a trending increase in neutralization at 2-6 weeks post-second dose. Non-spike-specific T-cells predominantly targeted SARS-CoV-2 non-structural protein at 6-months post-second dose in cross-reactive participants. mRNA vaccination was lastly shown to induce off-target T-cell responses against unrelated antigens. In summary, vaccine-induced spike-specific T-cell immunity appeared to influence serological neutralizing capacity, with only a modest effect induced by pre-existing cross-reactivity. IMPORTANCEOur findings provide important insights on the potential contributions of mRNA vaccine-induced spike-specific T-cell responses to the durability of neutralizing antibody levels in both uninfected and hybrid immune recipients. Our study additionally sheds light on the precise impacts of pre-existing cross-reactive T-cell immunity to SARS-CoV-2 on the magnitude and kinetics of cellular and humoral responses to vaccination. This study should ultimately inform the development of novel pan-coronavirus vaccines and vaccine regimens that can maximize the durability and breadth of protection against both current and future human coronaviruses of concern.

immunology↗

bioRxiv2024

Unveiling Tissue Structure and Tumor Microenvironment from Spatially Resolved Transcriptomics by Hypergraph Learning

Spatial omics technologies have revolutionized life sciences by enabling the simultaneous acquisition of biomolecular and spatial information. Identifying spatial patterns is crucial for understanding organ development and tumor microenvironments. However, the emergence of diverse spatial omics resolutions in these technologies has made it challenging to accurately characterize spatial domains at finer resolutions. To address this, we propose HyperSTAR, a hypergraph-based method designed to precisely identify spatial domains across varying resolutions by leveraging higher-order relationships among spatially adjacent tissue programs. Specifically, a gene expression-guided hyperedge decomposition module is introduced to refine the hypergraph structure to accurately delineate spatial domains boundaries. Additionally, a hypergraph attention convolutional neural network is designed to adaptively learn the importance of each hyperedge, enhancing the models ability to capture complex higher-order relationships within spatially neighboring multi-spots and/or single cells. HyperSTAR outperforms existing graph neural network models in tasks such as uncovering tissue substructures, inferring spatiotemporal patterns, and denoising spatially resolved gene expressions. It effectively handles diverse spatial omics data types and scales seamlessly to large datasets. The method successfully reveals spatial heterogeneity in breast cancer sections, with findings validated through functional and survival analyses of independent clinical data. HyperSTAR represents a significant advancement in spatial omics analysis, representing a robust tool for exploring complex spatial patterns across varying resolutions and data types. Its ability to capture intricate higher-order relationships among spatially neighboring spots/cells makes it an invaluable tool for advancing research in life sciences, particularly in cancer and developmental biology.

bioinformatics↗

bioRxiv2024

EF-P and its paralog EfpL (YeiP) differentially control translation of proline containing sequences

Polyproline sequences (XPPX) stall ribosomes, thus being deleterious for all living organisms. In bacteria, translation elongation factor P (EF-P) plays a crucial role in overcoming such arrests. 12% of eubacteria possess an EF-P paralog - YeiP (EfpL) of unknown function. Here, we functionally and structurally characterize EfpL from Escherichia coli and demonstrate its yet unrecognized role in the translational stress response. Through ribosome profiling, we analyzed the EfpL arrest motif spectrum and discovered additional stalls beyond the canonical XPPX motifs at single-proline sequences (XPX), that both EF-P and EfpL can resolve. Notably, the two factors can also induce pauses. We further report that, contrary to the housekeeping EF-P, EfpL can sense the metabolic state of the cell, via lysine acylation. Together, our work uncovers a new player in ribosome rescue at proline-containing sequences, and provides evidence that co-occurrence of EF-P and EfpL is an evolutionary driver for higher bacterial growth rates.

microbiology↗

bioRxiv2023

The Effects of Different Fatigue Types on Action Anticipation and Physical Performance in High-level Volleyball Players

This study examined the effects of different fatigue types on action anticipation and physical performance in high-level volleyball players. Twenty-four participants underwent four counterbalanced conditions: 60-min of cycling at 60% peak power output, 60-min Stroop task, 60-min cycling at 60% peak power output with Stroop task and 60-min neutral documentary to induce physical fatigue (PF), mental fatigue (MF), dual fatigue (DF) and control group (CG), respectively. Action anticipation (anticipation test and visual search test) and physical performance (countermovement jump, T-test, and spike test) were conducted at baseline, immediately after (Post1), and 40-min after fatigue intervention (Post2). DF and PF led to significantly lower jump height, T-test completion time, and spiking speed at Post1 and Post2 compared with CG and MF. Compared with PF, DF led to a significant decline in jumping, agility, and spiking performance at Post1 and decreased jumping performance at Post2. MF significantly decreased reaction time in the anticipation test at Post1 compared with CG. Reaction times in the visual search test were significantly lower with DF and MF at Post2 compared with CG. PF led to a greater decline in physical performance and this was exacerbated in DF, yet anticipation performance remained lesser impact.

physiology↗