Neuroscience Today

Cerebral noradrenergic activity modulates physiological functions of behaviour, cognition, movement, arousal and sleep. This review aims to provide an accurate summary of the current knowledge on the involvement of the noradrenergic system in Parkinson's Disease (PD) and its clinical correlations based on neuroimaging studies. Studies in PD highlight neuromelanin MRI signal loss in the locus coeruleus (LC), and positron emission tomography shows noradrenergic denervation across subcortical and cortical areas. More severe phenotypes of PD, manifesting with cognitive decline, apathy, REM sleep behaviour disorder and autonomic dysfunction, are associated with more severe noradrenergic dysfunction. Conversely more preserved noradrenergic transmission is common in tremulous PD. Furthermore, noradrenergic dysfunction, is also involved in transient motor manifestations such as tremor and freezing of gait. Recent neuroimaging advances greatly expanded the knowledge about noradrenergic dysfunction pathophysiology in PD. However, pharmacological treatment of its several associated manifestations is still lacking and needs further investigation.

Neurons active during both action execution and observation [mirror neurons (MirNs)] are central to theories of action understanding, yet what they represent remains debated: abstract goals, static grips, or movement kinematics? We recorded 433 neurons from macaque premotor cortex during execution and observation of reach-to-grasp actions. Population analyses revealed grasp-specific information in both conditions, broadly distributed across neurons and dynamically reconfigured over time. Generalization across task phases was limited, indicating time-specific and evolving population codes rather than static representations. Execution and observation were linked by a shared, partially overlapping population geometry that supported reliable cross-condition classification, with the strongest alignment emerging during movement and hold. Neural activity was systematically related to multidimensional hand kinematics, and these relationships generalized across neuronal populations and across agents. Together, these findings support a dynamic population-level account of MirN function in which premotor circuits integrate visual and motor signals to represent and anticipate the unfolding structure of others' actions.

In the human brain, dendrites exhibit nonlinear integration and sparse parallel processing capabilities, which can effectively perform visual tasks by integrating only a small subset of neuronal signals and play a crucial role in high-level information inference. However, conventional neuromorphic devices often ignore these important properties and require all neurons to perceive complete information. This makes it difficult to effectively replicate the efficient spatiotemporal processing capabilities of biological neuron dendrites. In this study, we present an artificial neuron dendrite array that integrates neurons, synapses, and dendrites, emulating the spatiotemporal spike integration properties of biological dendrites for precise parallel computation. Through multigate threshold regulation, the array enables parallel sparse spiking inference with random spatial distribution. This inference process forms a sparse dendritic spiking neural network (SD-SNN) that can perform compression, depth detection, and prediction. As a result, the SD-SNN achieves high-efficiency static and dynamic object processing while using only 0.5% of neuronal activity, slashing the power consumption by 98 and 65%, respectively. Our work reduces neural activity in the perception process by 99.5% while enhancing spatiotemporal computing capabilities and computational efficiency.

Proper regulation of arousal maintains the balance of rest and activity and enables appropriate responses to stimuli; its disruption is a hallmark of many neurodevelopmental disorders. Although transcriptional mechanisms of arousal control are well defined, the contribution of posttranscriptional processes such as alternative splicing remains unclear. Here, we identify a critical role for the microexon splicing regulator srrm3 in maintaining arousal homeostasis in zebrafish. srrm3 mutants exhibit persistent hyperarousal characterized by sleep loss, sensory hypersensitivity, and elevated behavioral and neuronal activity. We identify the cyclic adenosine monophosphate (cAMP)-cAMP-dependent protein kinase (PKA)-cAMP response element-binding protein (CREB) signaling axis as a central driver of mutant hyperarousal. Specifically, pharmacological inhibition of cAMP signaling rescues mutant hyperactivity and associated transcriptional changes whereas wild-type cAMP activation phenocopies the mutant. Down-regulation of immediate early genes and reduced CREB phosphorylation further suggest adaptation to sustained neuronal activation. These findings establish srrm3 -dependent microexon splicing as a key molecular layer of arousal regulation linking RNA-processing defects to neuromodulatory imbalance.

Dysregulation of RNA m 6 A modification has been implicated in Alzheimer's disease (AD), but the molecular mechanisms remain largely unclear. Here, we identified the presence of m 6 A on mitochondria-encoded messenger RNAs (mt-mRNAs) in the brain, with elevated levels correlated with amyloid- (A ) deposition. Under physiological conditions, cytosolic m 6 A-modified mt-Nd4 is recognized and degraded by the m 6 A reader protein YTHDF2, thereby preventing aberrant activation of the RIG-I-MAVS innate immune pathway in neurons. Under A -associated pathological conditions, YTHDF2 expression is markedly down-regulated in neurons, leading to the accumulation of m 6 A-modified mt-Nd4 in the cytosol. This accumulation triggers RIG-I-MAVS activation and type I interferon (IFN) responses. Neuron-derived IFN- then amplifies neuroinflammation by activating surrounding microglia through a paracrine mechanism. Furthermore, neuronal Ythdf2 deficiency exacerbates A -associated neuroinflammation and cognitive decline. Together, these findings reveal a previously unrecognized m 6 A/YTHDF2-dependent regulatory axis that links mitochondrial RNA metabolism to innate immune activation and neuroinflammation in A pathology.

The inflorescence meristem (IM) of flowering plants is a stem cell niche that is the source of aboveground organs such as stems and flowers. Although transcriptional profiling has elucidated some cell types within this tissue, the transcriptional dynamics of differentiation from stem cells into the diverse cell types in these organs remain unknown. We used single-nucleus RNA sequencing to characterize the transcriptional landscape of the IM, linking shoot stem cells to early differentiation cell types. Although our analysis of previously known domains uncovered the role of GH3 gene family members in meristematic activity and phyllotaxis, we also identified unknown transcriptional patterns, including early cortex patterning within developing primordia. Trajectory inference analysis revealed the dynamics of the S-G 2 -M cell cycle phases, as well as gene expression programs driving differentiation toward specialized cell types such as early primordia, cortex, cambium, xylem, and phloem. Collectively, our findings advance the understanding of the cell fate transcriptional dynamics shaping shoot organ development.

Autism spectrum disorder (ASD) is a neurodevelopmental disorder shaped by genetic factors such as copy number variations (CNVs) and immunological factors such as maternal infection. However, most studies on the development of genetic ASD have focused on neurological aspects, and the role of immunity in genetic ASD remains unclear. Here, we demonstrate increased T cells in the brains of 15q11-13 duplication ( 15q dup ) mice, which model a common CNV associated with ASD. Elevated CXCL16 in the brains of 15q dup mice promoted T cell infiltration, specifically of V 6 + T cells that produce IL-17A. Deletion of V 6 + T cells throughout development or treatment with neutralizing antibodies against V 6 or IL-17A increased social behavior in 15q dup mice. These findings suggest that immune dysregulation contributes to social behavior deficits in 15q dup mice, consistent with observations in maternal immune activation models, and may represent a potential target for interventions for ASD-associated differences in social behavior.

Macrophages in the meninges contribute to immune defense of the central nervous system (CNS), yet their site-specific origin and function remain poorly understood. Using an intravenous model of streptococcal meningoencephalitis in mice, we found bacteria predominantly in the leptomeninges and dura. Nevertheless, monocyte infiltration into the leptomeninges and parenchyma strongly correlated with disease severity. In the dura, infection triggered activation and loss of resident macrophages, followed by rapid engraftment of inflammatory monocytes that transiently replenished the macrophage niche. Under homeostasis, dural monocytes were supplied CCR2 independently from adjacent skull bone marrow. During infection, this local source was insufficient, necessitating recruitment from peripheral bone marrow. Infection further reshaped monocyte ontogeny, increasing monocyte-dendritic cell progenitor-derived monocytes, which expressed higher major histocompatibility complex class II levels and persisted in the brain alongside CD4 + T cells during resolution. Together, these findings reveal dynamic, compartment-specific remodeling of monocyte recruitment and differentiation across CNS borders during bacterial meningoencephalitis.

Post-stroke cognitive impairment (PSCI) is a common sequela that occurs after ischaemic stroke (IS). This study aimed to investigate whether miR-409-3p is related to PSCI. Patients with IS were divided into two subgroups: PSCI and post-stroke cognitive normality (PSCN). The plasma level of miR-409-3p was determined by RT-qPCR. The association between PSCI and miR-409-3p was evaluated through binary logistic regression and by analysing the correlation between miR-409-3p and the MoCA score. In mice with middle cerebral artery occlusion (MCAO), the effects of miR-409-3p on cognitive function were explored through mNSS score and Morris water maze. In OGD/R-induced SH-SY5Y cells, the effects of miR-409-3p on cell viability, apoptosis and neuronal inflammation were evaluated using CCK-8, flow cytometry and ELISA. The content of miR-409-3p in patients with IS and those with PSCI was both increased, and its content showed a significant negative correlation with the MoCA score. The binary logistic regression analysis showed that a high risk of PSCI was associated with miR-409-3p. In MCAO mice, inhibition of miR-409-3p can significantly reduce the mNSS score and shorten the escape latency in the Morris water maze. Further-more, in neurons induced by OGD/R, down-regulation of miR-409-3p can exert a significant protective effect on neurons, manifested by enhanced cell viability, reduced apoptosis rate and inhibition of the synthesis of inflammatory factors. We conclude that miR-409-3p is a risk factor associated with PSCI. In MCAO mice and neurons induced by OGD/R, inhibition of miR-409-3p significantly alleviated neurological deficits and suppres-sed neuronal apoptosis and neuronal inflammation.

Alzheimer's disease (AD) is the most common form of dementia, driven by complex interactions among aging-related biological changes, neuronal degeneration, mitochondrial dysfunction, and environmental factors. Despite extensive research, effective disease-modifying therapies remain unavailable. Increasing evidence highlights the gut-brain axis as an important contributor to AD pathogenesis, particularly through amyloid-producing gut microbes that promote immune activation, neuroinflammation, and cerebral amyloid accumulation. This review summarizes current evidence linking gut microbiota (GM) dysbiosis to AD, focusing on microbial metabolites, neuroinflammatory pathways, and microbiota-targeted therapeutic strategies. A systematic analysis of experimental and clinical studies reveals that altered gut microbial composition is associated with systemic and neuroinflammation, blood-brain barrier dysfunction, oxidative stress, and neuronal damage. Key microbial metabolites, including short-chain fatty acids and indole derivatives, exhibit neuroprotective effects by regulating immune responses, maintaining barrier integrity, and supporting neuronal energy metabolism; disruption of these metabolites may accelerate neurodegeneration. Microbiota-based interventions such as probiotics, prebiotics, dietary modification, and fecal microbiota transplantation show beneficial effects in preclinical models by restoring microbial balance and reducing neuropathological features, although clinical evidence in humans remains limited. Overall, current findings support a contributory role of gut dysbiosis in AD and suggest that targeting the GM may offer a promising complementary strategy for disease modification and future therapeutic development.

Post-stroke cognitive impairment (PSCI) significantly impacts patients' quality of life following a stroke. Its effects on white matter functional networks remain unclear. This study used resting-state functional MRI to examine alterations in white matter functional connectivity and spontaneous activity in this condition. Resting-state functional MRI data were acquired from PSCI (n = 21), non-PSCI (PSNCI, n = 16), and healthy subjects (HSs, n = 29). A clustering analysis was employed to identify white matter functional networks. Functional connectivity and the amplitude of low-frequency fluctuation within these networks were compared across groups. Correlation analyses assessed their relationships with neuropsychological scores. Sixteen stable white matter networks were identified. Both patient groups showed reduced functional connectivity between the inferior longitudinal fasciculus and anterior cingulum compared to HSs. PSCI patients also exhibited decreased functional connectivity between the anterior and posterior cingulum compared to HSs. Furthermore, the amplitude of low-frequency fluctuation in the posterior cingulum was lower in PSCI group than in PSNCI group. These altered metrics showed correlations with neuropsychological performance across multiple cognitive domains. This study identified alterations in functional connectivity and spontaneous activity within specific WM networks in PSCI, which were associated with cognitive performance. These findings suggest that white matter functional abnormalities may be involved in the neural mechanisms underlying PSCI and could inform future research on early identification.

Sepsis-associated encephalopathy (SAE) is a common complication of sepsis characterized by neuronal injury and cognitive impairment. However, its underlying mechanisms remain unclear. In this study, we investigated the role of miR-125b in sepsis-induced hippocampal injury. A cecal ligation and puncture (CLP) model was established in mice, and miR-125b was overexpressed in the hippocampus using lentiviral vectors. CLP-induced sepsis increased mitochondrial fission and neuronal apoptosis in the hippocampus, accompanied by p53 activation and reduced miR-125b expression. miR-125b overexpression suppressed mitochondrial fission markers, reduced neuronal apoptosis, and improved cognitive deficits. Mechanistically, miR-125b inhibited ROS accumulation and p53 activation, whereas p53 overexpression reversed these protective effects. Conversely, hippocampal knockdown of miR-125b with LV-anti-miR-125b aggravated mitochondrial fission and neuronal apoptosis, further supporting its protective role. These findings indicate that miR-125b protects against sepsis-induced hippocampal injury by regulating the ROS/p53 pathway.

Parkinson's Disease (PD) represents the second most prevalent neurodegenerative condition which leads to the progressive destruction of dopaminergic neurons in the substantia nigra through oxidative stress mechanisms. The research evaluated Gallic Acid (GA) as a natural polyphenol with proven antioxidant properties for its ability to protect cells from 1-methyl-4-phenylpyridinium (MPP )-induced neurotoxicity in SH-SY5Y dopaminergic cell models. The research used SH-SY5Y cells which received 1 mM MPP treatment alongside different GA concentrations (25, 50 and 100 M) for 24 and 48 h. The CCK-8 assay measured cell viability while flow cytometry evaluated apoptosis and SOD and MDA levels determined oxidative status through SOD and Catalase and NO measurements. The addition of MPP resulted in a 32.74% decrease in cell viability at 48 h while simultaneously decreasing SOD and Catalase and NO levels and increasing MDA levels. The addition of 25 M GA protected cells from damage by increasing their viability to 86.53% at 48 h and decreasing apoptotic cell numbers. Our results revealed that co-treatment with 25-50 M GA effectively mitigated oxidative damage by preventing the depletion of catalase and NO levels. Furthermore, GA successfully reduced lipid peroxidation; specifically, 25 M GA decreased MDA levels from 21.18 to 9.64 nM/mg protein at 48 h, thereby restoring the cellular antioxidant defense system against MPP + -induced oxidative stress. In conclusion, the present study demonstrates that GA exerts a significant neuroprotective effect in an in vitro PD model by modulating the endogenous antioxidant network and alleviating lipid peroxidation. By effectively reversing the depletion of crucial enzymes and reducing apoptosis, GA shows potential therapeutic efficacy against oxidative stress-associated neurodegeneration. These findings suggest that GA is a promising phytochemical candidate warranting further in vivo evaluation to clarify its long-term bioavailability and translational value.

RNA metabolic dysregulation is a key pathological mechanism underlying the onset and progression of Alzheimer's disease (AD), involving multiple aspects such as abnormal RNA splicing, loss of function in RNA-binding proteins, dysregulation of non-coding RNAs, and impaired nuclear-cytoplasmic transport. In recent years, the emergence of epigenome research has revealed the critical role of RNA chemical modifications in regulating RNA metabolism at the post-transcriptional level. N4-acetylcytidine (ac4C) is the only known RNA acetylation modification in eukaryotes and is specifically catalyzed by N-acetyltransferase 10 (NAT10). The ac4C modification is widely found in tRNA, rRNA, and mRNA, and by influencing RNA stability, translation efficiency, and ribosome assembly, it participates in various biological processes such as the cell cycle, differentiation, aging, and stress responses. In AD, the ac4C modification profile undergoes significant changes, involving GABAergic synapses, the PI3K-AKT signaling pathway, and various lncRNAs. Although indirect evidence from progeria and tumor models suggests that via -tubulin acetylation and intersect with AD pathology via the p53 pathway and regulation of autophagy, these mechanisms currently lack direct experimental validation in NAT10 may participate in axonal transport through -tubulin acetylation and intersect with AD pathology via the p53 pathway and autophagy regulation, these mechanisms currently lack direct experimental validation within the AD system. This article systematically summarizes the molecular basis and regulatory networks of ac4C modification, integrates existing evidence and unresolved questions regarding its role in AD, and explores its potential value as a diagnostic biomarker and therapeutic target, with the aim of providing guidance for future research in this field.

Subclinical impulsive-compulsive behaviors (s-ICBs) in Parkinson's disease (PD) are common, clinically relevant, and frequently underdiagnosed. Objective behavioral measures reflecting vulnerability to impulsive-compulsive behavior are lacking. We investigated whether oculomotor measures of inhibitory control are associated with s-ICBs in dopaminergically treated PD patients without clinically manifest impulse control disorders. Twenty-nine patients with PD (Hoehn and Yahr stages 1-2) and twenty age-matched healthy controls completed prosaccade and antisaccade eye-tracking tasks. Executive functioning, trait impulsivity, quality of life and s-ICBs were assessed. Group comparisons and association analyses were performed using nonparametric methods with false discovery rate correction. Exploratory mediation analysis examined relationships between levodopa equivalent daily dose (LEDD), s-ICBs severity, and express prosaccades. All assessments were performed in the ON-medication state. Compared with healthy controls, patients with PD showed a higher frequency of express prosaccades and prolonged antisaccade latencies. Within the PD group, express prosaccades were moderately to strongly associated with s-ICBs severity and LEDD and were also related to reduced quality of life. Mediation analysis revealed a significant statistical indirect effect of dopaminergic medication on express prosaccades through s-ICBs severity, indicating that behavioral symptoms statistically accounted for a substantial proportion of this association. These findings demonstrate a close relationship between oculomotor control, subclinical impulsive-compulsive behaviors, and dopaminergic treatment in PD. The results highlight the complexity of non-motor manifestations in PD and underscore the need for longitudinal and mechanistic studies to clarify their clinical significance.

Due to the spread of misinformation that some vaccines cause autism spectrum disorder (ASD), many parents report changes in their vaccination behavior following a diagnosis of ASD, putting their children at increased risk for preventable diseases. Our study aimed to determine the rate of vaccination-related concerns and refusal behaviors in parents of children with ASD and non-autistic developmental delays (non-ASD-DD) and to examine factors potentially associated with vaccine hesitancy and refusal. In our study, a questionnaire was distributed to all parents of children diagnosed with ASD and non-ASD-DD who attended outpatient check-ups over 3 months at the Child Psychiatry Clinic of the two large hospitals. Participants completed a structured questionnaire assessing self-reported vaccination behaviors before and after diagnosis, as well as separate Likert-scale items evaluating vaccine-related beliefs and attitudes. No parents declined participation, and all 154 eligible parents were included in the study. Among the respondents, 87.7% were mothers. The most common diagnoses were intellectual disability (41.6%) and ASD (31.2%). Reported vaccine refusal increased from 3.9% before diagnosis to 9.7% after diagnosis (p = 0.012). The main reason cited for hesitancy or refusal was the belief that vaccines had caused their child's neurodevelopmental condition. No independent associations were found between post-diagnosis vaccine hesitancy/refusal and parental education, income, source of vaccine information, or depression/anxiety scores. However, 88.3% of participants disagreed or strongly disagreed with the statement that vaccines cause ASD or developmental disorders. Parental concerns about vaccines persist after a diagnosis of neurodevelopmental disorders. Tailored education and communication strategies are essential to support informed vaccine decision-making in these families and to prevent refusal for both diagnosed children and their siblings. Persistent misinformation linking childhood vaccines-particularly the MMR vaccine-to autism spectrum disorder continues to influence parental attitudes toward vaccination. Parents of children with ASD have been shown to exhibit higher rates of vaccine hesitancy, especially regarding vaccination of younger siblings. Vaccine hesitancy and refusal were more frequently reported after the diagnosis of not only autism spectrum disorder but also other developmental delays, including intellectual disability and speech delay. Although concerns about vaccines causing developmental disorders were common, vaccine refusal remained relatively uncommon and no independent predictors of post-diagnosis hesitancy or refusal were identified, highlighting the complex nature of parental vaccination decisions.

Chronic subdural hematoma (CSDH) remains a delayed complication after aneurysm clipping. Quantitative evidence linking postoperative pneumocephalus to CSDH is limited. To evaluate the association of a normalized CT index-the Air-Brain Index (ABI)-and intracranial volume (ICV) with postoperative CSDH, with prespecified sex adjustment. Single center retrospective cohort of adults undergoing clipping. Day 1 CT underwent standardized segmentation to derive ABI (air/brain) and ICV (air + brain). Multivariable logistic regression included age and sex; sex stratified analyses and ROC curves assessed performance. Among 68 patients, 18 developed CSDH. Higher ABI was associated with CSDH in univariable analysis; however, after adjustment for age and sex, ABI was no longer significant, whereas older age and male sex remained independent predictors. Although ABI was not independently associated with CSDH after adjustment for age and sex, it demonstrated a significant univariable relationship and may serve as a descriptive postoperative marker of residual intracranial air burden for hypothesis-generating risk stratification.

The immunosuppressive tumor microenvironment (TME) serves as a central driver of bladder cancer (BCa) progression and prognosis. While its significance is widely acknowledged, the key cellular subsets that mediate this immunosuppressive state and their core regulatory genes remain incompletely understood. Key immunosuppressive cellular subsets and their signature genes were systematically identified using single-cell RNA sequencing (scRNA-seq) data from BCa samples. A prognostic risk model was then constructed via univariate and multivariate Cox regression analyses, based on bulk RNA-seq datasets and the identified signature genes. The role of SUSD2 was further validated in vitro using qRT-PCR, Western blot, immunofluorescence, proliferation, and invasion assays. Compared with adjacent normal tissues, BCa tissues showed significant enrichment of stromal cells (e.g., epithelial cells, fibroblasts). Among these stromal populations, the proportion of myofibroblast-like cancer-associated fibroblasts (myCAFs) was significantly increased in BCa tissues, and high myCAF infiltration was closely associated with poor patient prognosis. Pseudotime trajectory analysis confirmed that fibroblast differentiation in BCa shifts toward a terminal state (State 3), which is predominantly composed of myCAFs. A prognostic model established using myCAF-related signature genes (TMEM74B, ABCC9, FCMR, ALG9, SUSD2, and ETV7) exhibited stable predictive performance in both training and validation cohorts, with SUSD2 identified as a risk-related gene. In vitro experiments revealed that SUSD2 knockdown inhibited myCAF activation and extracellular matrix secretion, thereby attenuating its promotional effects on BCa cell proliferation and invasion. The TGF- receptor inhibitor SB-431542could reverse the facilitative effects of SUSD2 overexpression on tumor cell proliferation and migration. Our findings identify myCAFs as a core regulatory cellular subset and SUSD2 as a key molecule within the immunosuppressive TME of BCa. Additionally, SUSD2 may trigger the activation of the TGF- /Smad signaling cascade to induce myCAF activation, thereby accelerating BCa progression. These results provide novel potential targets and a theoretical basis for prognosis assessment and TME-targeted therapy in BCa.

The present study investigates the antennae and brain of the immature stages of the caddisfly Hydropsyche pellucidula (Andersen, and Klubnes 1834) using scanning and transmission electron microscopy as well as fluorescence microscopy. The larval antenna is unsegmented and bears two long, articulated trichoid sensilla and two large, non-articulated basiconic sensilla, all with an internal structure typical of mechanoreceptors. No changes were detected between the larval stages examined, but larval sensilla differ completely from the adult sensilla previously described in the same species, which include several chemoreceptors (e.g. trichoid, pseudoplacoid, pseudoplacoid, chaetoid, coronary, and styloconic sensilla). A preliminary account of larval brain anatomy reveals a small central body without columnar elements, mushroom bodies without calyces, and no antennal lobes in any larval stage. This condition differs markedly from the pupal brain, which-similar to the adult brain-shows a central body with a vague fan-shaped structure, mushroom bodies with small calyces, and well-structured antennal lobes containing few but relatively large glomeruli. Such dramatic change is similar to what occurs in other closely related holometabolous insects, such as Lepidoptera, but it is more pronounced, probably because of the ecological differences between larvae and adults, as also observed in other aquatic insects such as mosquitoes. The results of this research shed light on overlooked aspects of caddisfly biology. Moreover, they may enhance our understanding of the evolution of insect olfaction, since caddisflies are among the most important orders of aquatic insects and the closest relatives of Lepidoptera, a key model system in insect chemical ecology.

Prehospital stroke scales are widely used to identify patients with suspected stroke. The scales have shown potential to identify suspected large-vessel occlusion (LVO) for direct-to-thrombectomy triage, yet it remains unclear which scales perform best, and whether performance varies by patient characteristics. In this retrospective diagnostic accuracy study, we analysed 1150 consecutive adults admitted under the stroke code protocol at a primary stroke centre (January 2015-December 2017). The cohort included ischaemic stroke, transient ischaemic attack, intracerebral haemorrhage and mimics. We evaluated 55 scale-threshold combinations (46 scales) for detection of computed tomography angiography (CTA)-confirmed LVO, quantified by area under the receiver operating characteristic curve (AUC), sensitivity, specificity and predictive values. In our study population, LVO prevalence was 13.5%. AUC ranged from 0.586 (Hemiplegia) to 0.855 (FAST-ED). No scale significantly outperformed the continuous National Institutes of Health Stroke Scale (NIHSS, AUC 0.851) after Bonferroni correction, and none achieved both sensitivity and specificity above 80%. Scales combining cortical and motor items outperformed purely motor-based instruments. Accuracy was comparable between sexes but higher for left-hemispheric strokes (mean AUC: 0.82 vs 0.76) and patients aged 60 years (0.82 vs 0.77). None of the prehospital stroke scales were accurate enough to reliably detect LVO. Performance varied with stroke laterality, patient age and stroke severity-approaching chance levels at the extremes of severity and showing a consistent hemispheric bias. This highlights fundamental limitations of using clinical scales alone to detect LVO in real-world stroke code populations. Registration: ClinicalTrials.gov, https://clinicaltrials.gov/study/NCT05378490, NCT05378490.

Research on amnesia has been fundamental in establishing the role of the human hippocampus in memory. Even though other structures within the hippocampal-diencephalic-cingulate network also play a role in episodic memory, studies of hippocampal amnesia often ignore the importance of changes in this broader network. In a large cohort of patients (n = 38) with hippocampal damage due to autoimmune limbic encephalitis, we previously found that amnesia was predominantly explained by resting-state functional abnormalities across this network. Here, we examined the integrity of individual diencephalic nuclei and white matter pathways, and its relationship with memory function. We found atrophy in the mammillary bodies, and the anterior, laterodorsal, pulvinar, and dorsomedial thalamic nuclei. Atrophy was often as pronounced as that in the hippocampal formation. Diencephalic volumes predicted memory over and above any hippocampal/subicular subfield volume estimate. White matter was compromised within and beyond this network. Fornix integrity was linked to diencephalic and hippocampal volumes, but not to recollection/recall. We strongly advise caution in employing the term "focal hippocampal damage" in cognitive neuroscience, and highlight the need to study the significance of plausibly knock-on effects in specific diencephalic nuclei and white matter tracts within broader circuits.

Working memory (WM) tasks often require comparing remembered items to test displays, but little is known about how people selectively remove irrelevant information at test. Across three experiments, we used contralateral delay activity (CDA) to track WM load and examine selective removal. In Experiment 1, CDA amplitudes increased with set size even when only one item was probed, suggesting minimal removal based on spatial location. Experiment 2 ruled out spatial grouping by presenting items sequentially in the same location, yet more items were retained for larger set size. In Experiment 3, however, when items belonged to distinct mnemonic categories, CDA amplitudes at test were reduced, consistent with selective removal based on category relevance. Additionally, P3 old-new effects showed that decision speed and strength were influenced by the number of items maintained. Together, these results suggest that people selectively remove WM contents based on categorical relevance, not spatial cues, enabling more efficient memory-based decisions.

Low reproducibility in brain-behavior association studies has prompted calls for larger samples, but rigorous quality control (QC) of behavioral measures may also strengthen the robustness of observed associations. This study examined how varying levels of behavioral QC affect brain-behavior associations using delay discounting, a widely studied measure of immediate reward orientation. Data were drawn from the Adolescent Brain and Cognitive Development Study (n = 10,936), and delay discounting data were subjected to eight increasingly stringent QC levels. Associations between delay discounting and brain structural morphometry were evaluated in 15 bilateral a priori regions of interest (ROIs), and changes in effect size, significant associations, and multivariate behavioral prediction were quantified across QC levels. Greater QC stringency substantially amplified association strength, with effect sizes increasing by an average of 188.52% across ROIs, the number of statistically significant associations more than doubling, and multivariate prediction improving by 267%. Although higher QC levels reduced sample size by up to 65%, strengthening of associations plateaued at intermediate levels, suggesting asymptotic improvements in resolution. Overall, stringent behavioral QC substantially increased the magnitude of brain-behavior associations, suggesting this is a critical methodological consideration for improving reproducibility in cognitive neuroscience.

Octopamine (OA), a biogenic amine functionally similar to vertebrate norepinephrine, plays an important role in invertebrate neurophysiology. Previous reports of putative octopaminergic cells in the pond snail, Lymnaea stagnalis, have revealed inconsistencies, prompting our investigation that combined in situ hybridization chain reaction with traditional immunohistochemical methods. We mapped tyramine -hydroxylase (TBH) mRNA, the corresponding TBH protein, and the OA neurotransmitter. Approximately 40 neurons were labeled by all three methods and are presumably genuinely octopaminergic, including several neurons with previous electrophysiological evidence that OA is their neurotransmitter. Our results also revealed approximately another 40 cells that only showed evidence of mRNA and enzyme labeling, but no neurotransmitter, and another five to eight cells that only labeled for OA. Some of these results are likely explained by antibody cross-reactivity, but multiple TBH isoforms (some of which may not produce OA) or regulatory mechanisms that block TBH function also need to be considered. Overall, this study underlines the need for a more nuanced interpretation of neuroanatomical mapping and provides new insights into the organization of molluscan octopaminergic systems.

Validated prognostic tools are essential to advance drug development and clinical care in Alzheimer's disease, particularly as the field shifts toward the prevention of cognitive decline. While progress has been made in developing blood-based biomarkers for the early detection of amyloid pathology, amyloid positivity alone does not reliably predict progression to symptomatic disease. Digital markers can serve as complementary prognostic tools to inform early intervention strategies. Among digital markers, speech-based markers offer a scalable, non-invasive, and cost-effective approach to predicting and monitoring cognitive decline. However, the development of validated speech-based tools has been constrained by the lack of large, multilingual datasets with longitudinal sampling, deep phenotyping, harmonized clinical and biomarker data, and adequate representation of preclinical populations. SpeechDx is a 3-year, multinational, multilingual observational study (n = 2006) designed to address these gaps and accelerate the development of speech-based tools to inform early risk assessment, enable timely intervention, and guide personalized care.

Mating and other behaviors emerge during adolescence through the coordinated actions of steroid hormone signaling throughout the nervous system and periphery. In this study, we investigated the transcriptional dynamics of the medial preoptic area (MPOA), a critical region for reproductive behavior, using single-cell RNA sequencing (scRNA-seq) and in situ hybridization techniques in male and female mice throughout adolescence development. Our findings reveal that estrogen receptor 1 (Esr1) plays a pivotal role in the transcriptional maturation of GABAergic neurons within the MPOA during adolescence. Deletion of the estrogen receptor gene, Esr1 , in GABAergic neurons (Vgat+) disrupted the developmental progression of mating behaviors in both sexes, while its deletion in glutamatergic neurons (Vglut2+) had no observable effect. In males and females, these neurons displayed distinct transcriptional trajectories, with hormone-dependent gene expression patterns emerging throughout adolescence and regulated by Esr1. Esr1 deletion in MPOA GABAergic neurons, prior to adolescence, arrested adolescent transcriptional progression of these cells and uncovered sex-specific gene-regulatory networks associated with Esr1 signaling. Our results underscore the critical role of Esr1 in orchestrating sex-specific transcriptional dynamics during adolescence, revealing gene regulatory networks implicated in the development of hypothalamic-controlled reproductive behaviors.

High-speed volumetric imaging of the brain is essential for linking diverse cellular events to tissue-level functions. However, the brain's structural and dynamic heterogeneity-spanning microns to millimeters and milliseconds to hours-requires imaging techniques with tunable spatiotemporal resolution, flexible 3D sampling, and compatibility with targeted perturbations. Here, we present tunable Bessel beam two-photon fluorescence microscopy (tBessel-TPFM), a compact, low-cost, and versatile platform for intravital brain imaging across millimeter scale with subcellular resolution. tBessel-TPFM transforms slow 3D volume scans into fast 2D frame scans via an axially elongated Bessel focus, achieving acquisition rates ~100 fold faster and reduced motion artifacts compared with conventional TPFM. Exploiting its full tunability of the Bessel focus, we applied tBessel-TPFM for quantitative mapping of cerebral blood flow and neurovascular coupling in normal and ischemic stroke mice. Unlike existing Bessel focus generation methods, the axial center of tBessel-TPFM remains fixed at the objective focal plane during profile tuning. Leveraging this advantage, we integrated tBessel-TPFM with simultaneous 3D targeted optogenetic stimulation for volumetric neuronal connectivity mapping. We also tracked microglial process dynamics following single-cell laser ablation, revealing diverse neuroimmune responses across spatial and temporal scales. By combining high speed, deep penetration, tunable sampling, and multimodal perturbation, tBessel-TPFM empowers a broad spectrum of neurobiological investigations-from vascular physiology and functional connectivity to neuroimmune interactions.

The central nervous system (CNS) can effectively control body movements despite environmental changes. While much is known about adaptation to external environmental changes, less is known about responses to internal bodily changes. This study investigates how the CNS adapts to long-term alterations in the musculoskeletal system using a tendon transfer model in nonhuman primates ( Macaca fuscata ). We surgically relocated finger flexor and extensor muscles to examine how the CNS adapts its strategy for finger movement control by measuring muscle activities during grasping tasks. Two months post-surgery, the monkeys demonstrated significant recovery of grasping function despite the initial disruption. Our findings suggest a two-phase CNS adaptation process: an initial phase enabling function with the transferred muscles, followed by a later phase abandoning this enabled function and restoring a control strategy that, while potentially less conflicted than the maladaptive state, resembled the original pattern, possibly representing a 'good enough' solution. These results highlight a multi-phase CNS adaptation process with distinct time constants in response to sudden bodily changes, offering potential insights into understanding and treating movement disorders.

Colorectal cancer (CRC) is a prevalent and highly lethal malignancy. However, the molecular mechanisms underlying its progression remain incompletely understood. Therefore, this study aimed to investigate the role and mechanisms of circKPNA2 in CRC progression. We used various techniques, including RNA sequencing, functional assays, RNA pull-down, and mass spectrometry, to examine the expression and function of circKPNA2. Our findings indicate that circKPNA2 is significantly upregulated in CRC tissues and cell lines. It contributes to tumour growth by promoting cell proliferation, migration, and invasion. We found that circKPNA2 specifically binds to the RIN1 protein and increases its levels, thereby activating the Ras signalling pathway and facilitating CRC progression. Additionally, we identified that METTL3 regulates the expression of circKPNA2 through N6-methyladenosine (m 6 A) methylation, which in turn affects CRC cell proliferation, migration, and invasion. These results deepen our understanding of CRC biology and underscore the potential of targeting circKPNA2 as both a therapeutic target and a diagnostic biomarker to improve CRC diagnosis and treatment.

In people, magnetic resonance imaging (MRI) with high-resolution, high T2-weighted (T2w) contrast-balanced steady-state free precession (b-SSFP) pulse sequences and 3D fast spoiled gradient-echo (FSPGR) pulse sequences can improve visualization of cranial nerves (CNs) and associated osseous foramina. This prospective, observational study aimed to determine whether the addition of b-SSFP and FSPGR sequences improves confidence in the identification of canine CNs compared to the standard imaging protocol at 1.5 T. The head of 10 canine cadavers was imaged, including transverse T1-weighted (T1w) and T2w fast spin echo (FSE); sagittal T2w FSE; 3D Fast Imaging Employing Steady-State Acquisition with Cycling (FIESTA-C); and 3D Liver Acquisition with Volume Acceleration (LAVA). Board-certified radiologists rated their confidence in identifying CNs II-XII using a four-point scale. Observers evaluated standard sequences alone, then with the addition of transverse FIESTA-C, and finally with the inclusion of FIESTA-C and transverse LAVA. Scores between protocols were compared with the Wilcoxon matched-pairs signed-rank test. Interobserver agreement was measured with weighted kappa. Median score increased when adding FIESTA-C for all observers for nerves VII, VIII, IX, and X, and for two observers for nerves III and XI. Matched-pairs comparison revealed significant differences for nerves VII and VIII for all observers, and for nerves III, IX, X, XI, and XII for two observers. Further addition of LAVA generally did not improve confidence. Interobserver agreement was overall moderate to substantial, but slight to fair for some nerves and pulse sequences. Addition of a 3D b-SSFP sequence may improve identification of some CNs, especially VII and VIII, which are somewhat commonly affected by middle ear pathology in dogs.

The quality of stable long-term recordings from chronically implanted electrode arrays is essential for experimental neuroscience and brain-computer interfaces. This work uses scanning electron microscopy (SEM) to image and analyze eight 96-channel Utah arrays previously implanted in motor cortical regions of four subjects (subject H = 2242 days implanted, F = 1875, U = 2680, C = 594), providing important contributions to a growing body of long-term implant research leveraging this imaging technology. Four of these arrays have been used in electrolytic lesioning experiments (H = 10 lesions, F = 1, U = 4, C = 1), a recently developed electrolytic perturbation technique demonstrated compatible with continued neuroelectrophysiology using small direct currents. Previously, our group showed that electrolytic lesioning can be used as a technique to create regions of controlled neuron loss without significantly changing recording quality (Bray, Clarke et al., 2024). Here, by surveying physical damage such as biological debris and material deterioration, we show that electrolytic lesioning causes no statistically significant material damage to the implanted electrode arrays. In addition to surveying physical damage, such as biological debris and material deterioration, this work also analyzes whether electrolytic lesioning created damage beyond what is typical for these arrays. These findings also indicate that there are no statistically significant differences between the damage observed on normal electrodes versus those used for electrolytic lesioning, yielding no evidence that electrolytic lesioning significantly affects the material quality of chronically implanted electrode arrays. Finally, this work also includes the largest collection of single-electrode SEM images for previously implanted multielectrode Utah arrays, spanning 11 different intact arrays and one broken array. As the clinical relevance of chronically implanted electrodes with single-neuron resolution continues to grow, these images may be used to provide the foundation for a larger public database and inform further electrode design and analyses.

Parkinson's disease is a neurodegenerative disorder characterized by degeneration of nigrostriatal dopamine neurons and basal ganglia dysfunction, with sex differences in risk and progression that are not yet fully understood. Here, we investigated how peripubertal loss of gonadal hormones influences nigrostriatal integrity, striatal synaptic architecture, and motor behavior in male mice. Using a castration model, we show that early androgen deprivation recapitulates key neuropathological and behavioral features of Parkinsonian hypodopaminergia, including reduced tyrosine hydroxylase-positive neurons in the substantia nigra pars compacta, impaired motor performance on the vertical pole and rotarod, and disrupted forelimb kinematics. At the circuit level, androgen loss markedly reduced mushroom spine density on both direct- and indirect pathway striatal spiny projection neurons, with a concomitant increase in immature spine classes, consistent with impaired synaptic maturation. These structural and behavioral deficits were accompanied by reduced striatal Tyrosine receptor kinase-B (TrkB) protein, suggesting attenuation of brain derived neurotrophic factor (BDNF)/TrkB trophic signaling. Testosterone replacement preserved nigral dopaminergic neurons, normalized motor behavior, prevented mushroom spine loss across both striatal pathways, and selectively increased long/thin spines on direct-pathway neurons, suggesting enhanced synaptic plasticity. Together, these findings support a hormone-trophic model in which androgen signaling maintains nigrostriatal integrity and striatal synaptic maturation through androgen receptor-dependent BDNF/TrkB mechanisms. Although Parkinson's disease predominantly manifests with aging, these results suggest that early disruption of androgen signaling can produce lasting circuit-level alterations that influence vulnerability to degeneration over time, providing mechanistic insight into sex differences in Parkinson's Disease.

Tuberculous meningitis (TBM), the most severe form of Mycobacterium tuberculosis infection, is characterized by high mortality and neurological sequelae, largely attributed to blood-brain barrier (BBB) disruption. While recent studies identified GSDMD-mediated endothelial pyroptosis as a key mechanism of inflammatory BBB damage, the full molecular landscape in TBM remains unclear. This study employed an integrated multi-omics approach, combining bulk and single-cell RNA sequencing of clinical and murine datasets with experimental validation, to identify central mediators of BBB dysfunction in TBM. We identified and validated three genes - MARCKS, CD274 (PD-L1), and IL17RA - as significantly upregulated in TBM. Single-cell analysis of a murine TBM model demonstrated predominant MARCKS expression in CNS microglia, a finding further supported by elevated MARCKS protein levels in peripheral blood mononuclear cells from patients. However, validation in human TBM brain tissue remains warranted. Functional enrichment and correlation analyses positioned MARCKS at the nexus of inflammatory signaling and cytoskeletal regulation, showing strong associations with key effectors of the GSDMD pyroptosis pathway (CASP4, CD14, NINJ1). Our data further indicate a robust co expression pattern between MARCKS and key effectors of the GSDMD pathway (CASP5, TLR4, and CASP1), suggesting that MARCKS mediated cytoskeletal destabilization and GSDMD dependent lytic pore formation may act in concert to promote BBB disruption. Nevertheless, this proposed mechanistic interplay requires direct experimental verification. Our findings nominate MARCKS as a novel mechanistic hub linking neuroinflammation to barrier pathology in TBM, revealing potential therapeutic targets for adjunctive barrier stabilizing strategies.

Microglia are a key driver of neurodegenerative disease, orchestrating inflammatory signaling, metabolic stress responses, synaptic remodeling, and neuronal fate within the central nervous system (CNS). Among experimental models, the human microglial cell line, HMC3, is one of the most widely used models for mechanistic investigation and pharmacological screening of microglial dysfunction, particularly in neurodegenerative contexts. Nevertheless, a key question remains: how faithfully does HMC3 reflect human microglial biology? This review integrates current evidence on HMC3 cells, including their molecular and metabolic features, functional plasticity, and disease-oriented applications. HMC3 cells reproduce hallmark neurodegeneration-associated programs, such as stimulus-dependent polarization, oxidative and endoplasmic reticulum stress signaling, inflammasome activation, autophagy dysregulation, lipid remodeling, angiogenic cross-talk, and phagocytic clearance of amyloid and apoptotic debris, modeling processes relevant to Alzheimer's disease, Parkinson's disease, ischemic injury, and metabolic neurodegeneration. Neuron-microglia co-culture systems further demonstrate the direct impacts of HMC3 activation states on neuronal vulnerability and survival. We also summarize the expanding repertoire of pharmacological and genetic interventions applied to HMC3, highlighting their compatibility with high-throughput and multi-omics discovery platforms. Despite inherent limitations of immortalized models, HMC3 represents a powerful front-line tool for dissecting neurodegenerative microglial mechanisms and steering early therapeutic discovery.

Johne's disease (JD), caused by Mycobacterium avium subsp . paratuberculosis (MAP), is a chronic enteric disease affecting cattle worldwide, with considerable economic implications for dairy producers. MAP is a robust, intracellular pathogen highly resilient to environmental challenges. To elucidate the molecular mechanisms underlying MAP infection in desi cattle ( Bos indicus ), we performed a comprehensive transcriptomic analysis. Diseased and healthy female desi cattle were selected for RNA sequencing based on phenotype. Raw data were processed, and differential expression of mRNA and lncRNA was determined. Gene modules and hub genes significantly correlated with traits were identified using network analysis. Further analysis included functional enrichment, PPI network construction, and lncRNA-mRNA co-expression within the key module. The differential expression of final hub genes was validated via qPCR. RNA sequencing revealed 1,905 differentially expressed protein-coding genes and, for the first time, comprehensively annotated 45,947 lncRNA genes in desi cattle, of which 3,123 lncRNAs were differentially expressed in MAP-infected samples. Weighted Gene Co-expression Network Analysis identified 11 co-expressed gene modules, among which the turquoise module, comprising 870 protein-coding genes and 934 lncRNAs, was highly correlated with clinical traits. Functional enrichment analysis of this key module revealed significant involvement in defense response, inflammatory processes, and natural killer cell-mediated immunity and cytotoxicity. Gene set enrichment analysis highlighted suppressed pathways related to natural killer cell immunity, lectin response, and protein-DNA complex assembly, while activated pathways included G-protein-coupled receptor signaling, circulatory system processes, and metabolic pathways. Using network-based approach, we identified 12 hub genes (IL7R, TLR4, KLRK1, IFNG, TGFB1, CD68, CXCR6, GZMB, KLRG1, MMP9, GZMA, and SELL) in the turquoise module associated with MAP infection in PBMC transcriptomes across a mixed-breed Bos indicus cohort. These genes were associated with immune suppression and evasion, inflammation, and tissue remodeling. Additionally, 22 differentially expressed lncRNAs co-expressed with hub genes were identified, suggesting potential roles in modulating immune response. Our findings provide novel insights into the molecular mechanisms of MAP infection in desi cattle, identifying critical hub genes and lncRNAs as potential targets for developing innovative diagnostic and therapeutic strategies for Johne's disease.

Prostate cancer is one of the most common malignant tumors of the male genitourinary system. The impaired activity of natural killer (NK) cells observed in prostate cancer may contribute to immune evasion. This study aimed to develop robust NK cell-related diagnostic signatures. Based on NK related genes identified by scRNA-seq analysis, weighted gene co-expression network analysis, least absolute shrinkage and selection operator regression analysis, and machine learning algorithms were used to develop a novel diagnostic model. The expression of diagnostic genes was validated using tumor and adjacent normal tissues collected from prostate cancer patients. The biological functions of KIT as an NK cell related gene were further evaluated in prostate cancer cells. A nine-gene NK cell-related diagnostic signature was developed, including HSPD1 , HSPE1 , CLU , KIT , LAPTM4A , SLC18A2 , TUBA4A , VWA5A , and ZFP36L1 . These genes were validated in independent datasets and showed strong predictive ability for prostate cancer diagnosis (AUC 0.8). Based on the expression profiles of these genes, nine compounds were identified that may influence drug sensitivity in prostate cancer. Furthermore, the expressions of CLU , TUBA4A , and KIT were successfully validated in collected prostate cancer tissue samples. Functional experiments demonstrated that KIT overexpression enhanced the cytotoxicity of NK-92 cells against prostate cancer PC-3 cells, inhibited cancer cell viability, cell migration and invasion, and increased the secretion of cytokines such as IFN- , Gzms-A, Gzms-B, and Perforin, as well as the degranulation marker CD107a. This study provides a new understanding of NK cell related gene signatures in prostate cancer diagnosis and highlights KIT as a promising candidate for future therapeutic investigation. Further research is needed to explore the mechanisms underlying the expression of these genes and their roles in the tumor microenvironment.

Traumatic brain injury (TBI) is increasingly recognized as a systemic disorder with consequences that extend beyond the central nervous system (CNS) to include clinically relevant cardiac dysfunction. Clinical and experimental evidence indicate that TBI is associated with arrhythmias, myocardial injury, autonomic imbalance, and impaired cardiac performance, even in the absence of primary cardiac disease. These observations support the concept of a brain-heart axis through which neural injury influences cardiovascular regulation. Current evidence shows that multiple interacting mechanisms contribute to cardiac abnormalities following TBI. Acute autonomic dysregulation, particularly sympathetic overactivation, is strongly supported as an early driver of cardiovascular instability, while neuroimmune and neuroendocrine responses may modulate the persistence and severity of downstream effects. Emerging evidence further suggests that cerebrovascular dysfunction may act as a modifier of neurocardiac vulnerability, although direct clinical evidence remains limited. At the myocardial level, processes including inflammatory signaling, oxidative stress, and mitochondrial dysfunction are associated with electrophysiologic instability and impaired contractile function. However, much of the mechanistic understanding derives from experimental models, and translation to human populations remains incomplete. Accordingly, cardiac involvement after TBI is best understood as a multifactorial and heterogeneous process rather than the consequence of a single dominant pathway. This review synthesizes current clinical and experimental evidence within an integrated framework, emphasizing the interplay among autonomic, immune, and vascular mechanisms. Improved understanding of these interactions may enhance risk stratification and support the development of targeted strategies to mitigate cardiovascular complications after TBI.

Salmonella is a major foodborne pathogen that causes approximately 1.35 million infections annually in the US and remains a leading cause of poultry-associated foodborne illness. To improve chickens' resistance to this pathogen, it is important to understand the mucosal immune mechanisms that govern intestinal defense. Intraepithelial lymphocytes (IELs) positioned between intestinal epithelial cells provide frontline immune surveillance against enteric pathogens. However, a comprehensive characterization of IEL subtype responses to Salmonella infection remains incomplete. Therefore, we conducted this study to examine IEL subtypes and their mechanisms in response to a Salmonella enterica serovar Enteritidis ( S. Enteritidis) challenge using a combination of spectral flow cytometry and single-cell RNA sequencing (scRNA-seq). Fifty specific-pathogen free (SPF) chicks were reared to 21 days of age and then assigned to S. Enteritidis-challenged (SE; 1.62 10 8 CFU/bird, oral gavage) or control (CN; PBS) groups (n = 25/group). On day 2 post infection (2 dpi) and 6 dpi, eight birds per group were sampled to collect liver and ceca for bacteriology and ileum for IEL acquisition. Bacteriological findings confirmed the challenge: the SE group harbored S. Enteritidis at both time points. Flow cytometry results showed that Salmonella challenge increased the proportion of TCR + CD8 + cytotoxic IELs at 2 dpi, as well as the overall IEL proportion at 2 and 6 dpi. Notably, scRNA-seq identified clusters of progenitor T cells that significantly expanded and innate-like cytotoxic T cells, which emerged in SE-challenged birds at 2 dpi, indicating rapid mobilization of an innate-like cytotoxic response. Integration of flow cytometry and scRNA-seq data provided evidence that cytotoxic T cells expressing CD8 acquire innate-like transcriptional signatures within the intestinal epithelial compartment, suggesting functional reprogramming that enables rapid antigen responses. Trajectory analysis identified a robust transcriptionally inferred trajectory from progenitor T cells through activated CD8 + T cells to innate-like CTL as the predicted terminal cluster, with quiescent stem-like resident memory T cells transcriptionally positioned as a reservoir. These findings reveal a previously uncharacterized innate-like cytotoxic IEL response as a critical early defense mechanism against Salmonella in poultry and identify self-renewing stem-like Trm cells as a reservoir for rapid IEL effector differentiation.

Colorectal cancer (CRC) exhibits substantial biological and prognostic heterogeneity that is not fully captured by conventional clinicopathological staging, and the interplay between the cellular senescence and circadian dysregulation in CRC remains insufficiently defined. We integrated bulk transcriptomic and clinical data from public CRC cohorts to construct a senescence-circadian interplay score (SCore). Senescence-related and circadian-related gene sets were intersected with CRC differentially expressed genes to identify candidate genes, from which a four-gene random survival forest model was established. The model was evaluated in the TCGA-COAD/READ training cohort and externally validated in GSE12945 and GSE39582. We further characterized clinicopathological associations, prognostic independence, nomogram performance, pathway enrichment, consensus molecular subtype distribution, immune landscape features, and predicted drug sensitivity. Single-cell RNA sequencing dataset was used to localize SCore-associated programs and to examine T-cell communication and differentiation dynamics. In addition, RT-qPCR in CRC cell lines, immunohistochemical validation in 120 paired CRC and adjacent non-tumor tissues, and NOX4-centered knockdown experiments were performed to provide orthogonal experimental support. A higher SCore was consistently associated with poorer overall survival across cohorts and retained prognostic value when integrated with clinicopathological variables. High-SCore tumors were characterized by enrichment of oxidative stress, extracellular matrix remodeling, focal adhesion, and invasion-related programs, together with computationally inferred immune dysfunction/exclusion-associated features. Single-cell analyses localized SCore-associated signals to a T-cell-centered context, where cell-cell communication, pseudotime dynamics, and functional-state remodeling converged. Notably, MIF-(CD74+CXCR4) signaling emerged as a prominent interaction axis. Consistent with the transcriptomic findings, immunohistochemistry confirmed higher expression of NOX4, CXCL1, CDKN2A, and SIX1 in CRC tissues than in paired adjacent non-tumor tissues. Functionally, NOX4 knockdown reduced intracellular ROS, attenuated multiple inflammatory/immunoregulatory mediators, lowered PD-L1 protein expression, and suppressed migratory capacity, supporting a NOX4-associated redox/inflammatory component within the broader SCore-linked state. SCore provides biologically interpretable transcriptomic framework for CRC risk stratification. The NOX4-centered data provide functional support for one component of this state, whereas the broader immune and circadian implications remain hypothesis-generating.

Chaperone-mediated autophagy (CMA) plays an important role in tumor progression and remodeling of the tumor immune microenvironment. However, its functional heterogeneity, immune associations, and clinical significance in colon cancer remain unclear. Single-cell and bulk transcriptomic data were integrated to characterize CMA-related features in colon cancer. At the single-cell level, CMA activity was assessed across cell types, and differentially expressed genes between CMA-high and CMA-low groups were identified in myeloid subpopulations. Robust candidates were screened by recurrence frequency. At the bulk level, TCGA-COAD was used as the primary training cohort, and GSE17538 and GSE38832 for cross-cohort performance evaluation. CMA activity was quantified by ssGSEA, and candidates were further refined by WGCNA and tumor-normal differential expression analysis. An ensemble machine learning framework incorporating 101 algorithm combinations was used to construct a dual prognostic system consisting of the Risk Score and the Immune Risk Score. Prognostic performance was evaluated by Kaplan-Meier analysis, time-dependent ROC curves, and Cox regression. The Immune Risk Score was additionally evaluated in an independent single-center transcriptome cohort from Liaoning Central Hospital. MAPKAPK3 was identified as a key functional gene and validated in vitro . Drug sensitivity was predicted using pRRophetic and CGP2016. CMA activity showed marked intercellular heterogeneity and was predominantly enriched in myeloid cells. Frequency-based screening, WGCNA, and tumor-normal differential expression analysis identified robust CMA-related candidates. The Risk Score showed favorable prognostic performance and generalizability across cohorts. The Risk Score remained an independent prognostic factor, whereas the Immune Risk Score functioned as an integrated prognostic score combining clinicopathologic and immune microenvironmental information. Immune analyses revealed consistent differences in regulatory T cells and resting dendritic cells across risk groups, suggesting an association between CMA-related risk states and an immunosuppressive microenvironment. The three-variable clinicomicroenvironmental model showed good predictive performance. MAPKAPK3 overexpression promoted proliferation, migration, and invasion of colon cancer cells, providing preliminary gain-of-function evidence for its tumor-promoting role. This study revealed CMA-related heterogeneity and immune microenvironmental features in colon cancer and established a robust dual prognostic system. MAPKAPK3 may serve as a key functional gene associated with tumor progression and microenvironment remodeling within the CMA-related prognostic framework.

Epithelial barrier disruption and mucosal innate immune activation reinforce each other in ulcerative colitis (UC), yet the epithelial glycosylation programs linking mucus homeostasis to inflammatory tissue injury remain incompletely defined. Here, we integrated bulk, single-cell, and spatial transcriptomic analyses with functional validation to identify mucin-associated epithelial determinants relevant to UC. In the discovery cohort GSE107499, 15 dysregulated mucin-type O-glycosylation and mucin-associated genes were identified and were enriched mainly in O-glycan biosynthesis, glycosyltransferase activity, and Golgi-associated secretory functions. An integrative prioritization framework highlighted GALNT12 as the leading candidate. Low GALNT12 expression was associated with enrichment of TNFA signaling via NF-kB, inflammatory response, interferon gamma response, IL6-JAK-STAT3 signaling, and complement, together with higher infiltration scores for monocyte lineage cells, neutrophils, fibroblasts, and T-cell populations. At single-cell resolution, GALNT12 was concentrated in epithelial cells, particularly MUC2-positive epithelial cells. MUC2-positive epithelial cells with detectable GALNT12 expression showed broader inferred incoming and outgoing communication with surrounding epithelial, stromal, and immune populations and were enriched for proteostasis and secretory trafficking programs. Spatial transcriptomic analysis further identified GALNT12-high mucin-associated niches that co-localized with stronger goblet and mucin signatures and lower inflammatory scores. In HT29-19A cells, GALNT12 knockdown exacerbated TNF- -induced LDH release, reduced OCLN and TJP1 expression, increased IL-8, IL-6, CCL2, IL-1 , and IL-18 production, and enhanced caspase-1 processing detected as cleaved caspase-1 p10 together with increased GSDMD-N formation under TNF- stress. Collectively, these findings identify a GALNT12-centered mucin-associated epithelial program linked to epithelial barrier resilience and relatively restrained innate inflammatory injury at the barrier-immune interface in UC, while supporting a caspase-1/GSDMD-associated inflammatory injury phenotype under cytokine stress.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies and is characterized by pronounced phenotypic plasticity, metabolic adaptation, and therapeutic resistance within a dense and desmoplastic tumor microenvironment. Although transcriptional deregulation has been extensively investigated, post-transcriptional regulation, particularly the control of mRNA stability, has emerged as a critical and previously underexplored contributor to PDAC progression. RNA-binding proteins (RBPs), together with cis-regulatory RNA elements and epitranscriptomic modifications such as N6-methyladenosine (m6A), form interconnected regulatory networks that dynamically modulate mRNA turnover and thereby shape protein output in response to microenvironmental stress. By selectively stabilizing transcripts encoding epithelial-mesenchymal transition (EMT) regulators, metabolic enzymes, and stress-response factors, these networks promote reversible, non-genetic adaptation without requiring permanent genetic alterations. This regulatory flexibility supports invasion, therapeutic tolerance, and intratumoral heterogeneity under hypovascular and nutrient-limited conditions. Recent advances further suggest that targeting mRNA stability through small molecules and RNA-directed strategies may provide new therapeutic opportunities in PDAC. In this review, we summarize current insights into post-transcriptional mechanisms regulating mRNA stability in PDAC, highlight key knowledge gaps, and discuss their potential translational implications.

Risperidone is a widely used second-generation antipsychotic for schizophrenia, known for its clinical efficacy but also for long-term metabolic side effects, including weight gain and glucose-lipid dysregulation. Although its therapeutic effects have been well characterized, the molecular basis linking its benefits and adverse metabolic outcomes remains poorly understood. To identify genes mediating both therapeutic and metabolic responses to risperidone, we conducted an integrative multiomics study combining large-scale genetic association data with transcriptomic and epigenetic profiling. Differentially expressed genes following risperidone exposure were intersected with schizophrenia- and metabolism-associated loci using summary data-based Mendelian randomization analysis based on GWAS summary statistics and eQTL data to uncover germline variants, germline susceptibility, and potential germline alteration. DNA methylation profiling from patient-derived peripheral blood mononuclear cells (PBMCs) was used for regulatory validation. We identified 120 genes significantly modulated by risperidone, among which DKK3, EEF1A1, and PRKAA1 were causally associated with schizophrenia and metabolic traits through germline mutation-related regulatory evidence. Notably, DKK3 was downregulated after risperidone exposure and exhibited promoter hypermethylation, consistent with epigenetic regulation interacting with germline alteration and germline susceptibility. Functional correlation analysis revealed that lower DKK3 expression was associated with glycolipid dysregulation, supporting its role as a molecular bridge between antipsychotic action and metabolic liability. Our findings identify DKK3 as a germline mutation-related and epigenetically regulated candidate that bridges risperidone's neuropsychiatric benefits with its metabolic risks. This work offers novel insight into the shared molecular basis of antipsychotic response and side effects and suggests DKK3 as a promising biomarker for personalized treatment strategies in schizophrenia informed by germline regulatory variation and potential germline alteration.

Colorectal cancer (CRC) is shaped by inherited genetic susceptibility, metabolic reprogramming, epigenetic regulation, and tumor microenvironment (TME) heterogeneity. Lactylation has recently emerged as an epigenetic mechanism that links lactate accumulation to chromatin remodeling and transcriptional regulation. However, the roles of lactylation-related genes in CRC initiation and progression, particularly from the perspective of SNP-based germline genetic variation, remain to be elucidated. We integrated GWAS analysis and single-cell RNA sequencing to identify lactylation-related pro-oncogenic genes with potential genetic causal relevance and to further characterize their cellular localization and microenvironmental implications in CRC. GWAS uses inherited germline genetic variants, particularly SNPs associated with gene expression, as instrumental variables, thereby reducing confounding and reverse causation compared with conventional tumor expression-based analyses. GWAS analysis has identified germline variants in AXIN1, FASN, MLH1, and RAD50 that genetically predicted increased CRC risk. Single-cell analysis was subsequently performed to evaluate the cell-type-specific distribution and functional relevance of these GWAS-identified genes within CRC tissues. Among them, AXIN1 exhibited clearer differential expression and cell-type-specific distribution in CRC-associated cell populations. AXIN1-high cell populations were associated with metabolic adaptation, proliferative activity, lactylation-related transcriptional programs, inflammatory responses, immune regulation, extracellular matrix remodeling, and TME adaptation. Besides, FASN was also confirmed as a genetically supported risk gene implicated in CRC metabolic reprogramming. MLH1 and RAD50 were retained as candidate risk genes at the germline causal level. This study integrates germline variant-based causal inference with single-cell microenvironmental interpretation, highlighting germline variant-regulated lactylation in promoting colorectal cancer risk.

This study examined the relationship between body composition and cardiac autonomic nervous system activity in educators. The objective was to explore the associations between body fat mass (BFM) and fat-free mass (FFM) with different heart rate variability (HRV) indices in a sample of educators and to assess whether FFM moderates the relationship between BFM and autonomic function. A descriptive cross-sectional study was conducted with 253 educators from compulsory and higher education institutions during the 2022-2023 academic year. Autonomic regulation was assessed via HRV, and body composition was measured using bioelectrical impedance analysis. Individuals with higher fat mass exhibited less favorable autonomic modulation, evidenced by lower values of the root mean square of successive differences (RMSSD), a time-domain index that reflects parasympathetic activity and the recovery capacity of the autonomic nervous system. Those with greater fat mass also showed a higher mean heart rate, whereas participants with higher FFM displayed more efficient cardiac activity, with lower mean, minimum, and maximum heart rates. In addition, FFM partially moderated the relationship between BFM and mean heart rate. By contrast, a higher total body water content was associated with a shift toward sympathetic dominance, reflected in higher low-frequency (LF) components-which represent a mixture of sympathetic and parasympathetic activity-and lower high-frequency (HF) components, which predominantly reflect parasympathetic activity. Conversely, individuals with higher FFM presented lower mean, minimum, and maximum heart rates, suggesting more efficient autonomic cardiac regulation. Overall, the findings demonstrate that body composition-particularly FFM and BFM-significantly influences HRV and autonomic nervous system modulation in educators, underscoring the importance of considering body composition in the assessment of autonomic function and in the design of personalized health strategies.

Partial sleep deprivation (PSD) poses health risks to the night-shift workers (NSW), but its underlying impacts on local brain function remain underexplored. Brain entropy (BEN), a nonlinear dynamic metric, has emerged as a novel parameter of choice in probing the temporal irregularity of brain activity, thus may offer new insights into the characterization of the brain dysfunction following sleep deprivation. Seventy-eight female medical NSWs and 30 non-NSW healthy controls (HC) were recruited in this study. Psychomotor vigilance tasks (PVT) and resting-state fMRI (rs-fMRI) were sequentially conducted at three conditions for NSWs: baseline (prior to NSW), PSD (immediately after NSW), and recovery (after 3-5 days of regular sleep). Nine NSWs were excluded due to consecutive or intermittent sleep duration exceeding 6 h on the night-shift day, or significant head motion in the rs-fMRI scans, resulting in 69 NSWs in the following analysis. Static and dynamic sample entropy (SampEn) metrics were calculated for BEN quantification. An L1-regularized logistic regression (LR) model was constructed based on baseline SampEn metrics to distinguish PSD-vulnerable (N = 35) and PSD-resistant (N = 34) individuals with their PVT performances as the classification reference. Compared to HCs, SampEn of young female NSWs reduced mainly in the occipital and temporal cortices, and associated with poor sleep quality. But these inter-group differences did not survive the strict multiple comparison correction. Both static and dynamic SampEn metrics of NSWs were significantly altered following acute PSD, and largely returned to the baseline after sleep recovery. The SampEn-based LR model achieved a classification accuracy value of 78.26% to distinguish PSD-vulnerable and PSD-resistant individuals. One-night of acute PSD in shift work leads to reduced but largely reversible BEN, whereas long-term occupational chronic PSD tends to reduce the BEN in distributed primary cortices and correlates with the poor sleep quality in young female NSWs. The BEN metrics could serve as a potential biomarker to identify individual susceptibility to PSD, which may contribute to the optimization of rotating-shift schedules.

Working memory (WM) deficits are frequently observed in patients with insomnia disorder (ID), but their neural basis is unclear. Glymphatic dysfunction and disrupted structural-functional coupling have been implicated, yet they have rarely been examined together, particularly in clinical populations. We conducted a multimodal MRI study in 391 ID patients. Glymphatic function was estimated using the diffusion tensor image analysis along the perivascular space (DTI-ALPS). The SFC was derived by correlating structural connectivity and functional connectivity. WM was measured by the longest span on the digit span backward task. Partial correlations and mediation analyses were performed to examine associations among sleep quality (Pittsburgh Sleep Quality Index, PSQI), DTI-ALPS, SFC, and WM performance. DTI-ALPS was negatively correlated with PSQI ( r = -0.17, p = 0.006), indicating reduced glymphatic clearance with poorer sleep quality. Global SFC was positively associated with DTI-ALPS ( r = 0.32, pFDR 0.001), but not with WM ( r = 0.01, p = 0.84). At the network level, SFC within the subcortical network (Sub-SFC) correlated with both DTI-ALPS ( r = 0.29, pFDR 0.001) and WM performance ( r = 0.28, pFDR 0.001). Mediation analysis revealed that DTI-ALPS and Sub-SFC jointly mediated the association between PSQI and WM performance, with a significant indirect effect (indirect effect = -0.074). This study provides novel evidence that impaired glymphatic clearance and reduced Sub-SFC form key neural pathways linking poor sleep quality to working-memory deficits in ID, and that DTI-ALPS and Sub-SFC may serve as useful biomarkers of cognitive vulnerability.

Schizophrenia (SCZ) and bipolar disorder (BD) are severe psychiatric conditions with overlapping clinical presentations, genetic risk factors, and brain network dysfunction. Whether alterations in large-scale intrinsic brain networks reflect shared or disorder-specific genetic influences remains poorly understood. Clarifying this distinction is essential for refining etiological models and improving diagnostic precision. Genome-wide inferred statistics (GWIS) were applied to decompose the genetic architecture of SCZ and BD into shared and unique components. Using resting-state network (RSN) data from the UK Biobank, functional connectivity (FC) and structural connectivity (SC) were extracted as neuroimaging phenotypes. Causal inference approaches were subsequently employed to infer potential directional relationships between brain network connectivity and each disorder. Analyses revealed both common and distinct patterns of brain network connectivity associated with SCZ and BD. Notably, SC within the default mode network (DMN) exhibited opposing effects across the two disorders, suggesting divergent structural underpinnings despite clinical overlap. Additionally, SC within the limbic network (LN) and frontotemporal control network demonstrated potential causal relationships with both conditions, implicating these circuits astransdiagnostic neural substrates. These findings illuminate the shared and disorder-specific genetic and neural architecture underlying SCZ and BD. Integrating genome-wide genetic methods with large-scale neuroimaging data offers a powerful framework for disentangling psychiatric comorbidity and may inform more targeted diagnostic criteria and individualized treatment strategies.

Disrupted structural connectivity is recognized as a key pathophysiological feature of schizophrenia (SCZ). However, the relationship between cortical similarity network alterations and gene expression remains poorly understood. We applied the Morphometric INverse Divergence framework to T1-weighted MRI from 1,216 participants. Cortical similarity networks were constructed, and global and nodal metrics were computed. Case-control comparisons were performed using linear models. Partial least squares (PLS) regression identified genes associated with spatial patterns of network alterations using Allen Human Brain Atlas data. Among global metrics, only the rich-club coefficient differed between groups, with a negligible effect size. Nodal metrics showed reduced eigenvector centrality and k-coreness centrality in left temporal/insular (somatomotor), lateral occipital (visual), anterior cingulate (salience/ventral attention), and posterior cingulate (default mode) regions. Participation coefficient was widely reduced in SCZ. For each nodal metric, we identified PLS2-positive and PLS2-negative gene sets. Across degree, eigenvector centrality, and k-coreness centrality, PLS2-negative genes were enriched for metal ion transport, linked to manic and nonorganic psychosis, and upregulated in adolescence and early adulthood. PLS2-positive genes were enriched for neuron projection development and learning or memory, but not psychotic disorders. These findings highlight synaptic and neurodevelopmental mechanisms underlying structural dysconnectivity in SCZ.

Scarring alopecia remains a major clinical challenge, characterized by irreversible hair follicle (HF) loss and replacement by scar tissue. Wound-induced hair follicle neogenesis (WIHN) serves as a model in which HFs can regenerate following skin wounding. In this study, we characterized the cellular landscape associated with differences in regenerative capacity between small wound (SW) and large wound (LW) tissues at a late regenerative stage (post-wounding day 22, PWD22), a time point at which newly formed hair follicles are already present. Using single-cell RNA sequencing (scRNA-seq), we profiled re-epithelialized wound tissues from SW and LW conditions and identified major cutaneous cell populations, including keratinocytes and fibroblasts. Keratinocytes were further categorized into 8 subpopulations, among which infundibular basal keratinocytes (INFU basal KCs) were increased in LW compared with SW tissues, as validated by immunofluorescence (IF) staining. Pseudotime analysis indicated differences in the temporal expression pattern of the INFU basal KC marker Fst between SW and LW conditions. Cell-cell communication between keratinocytes and fibroblasts showed that extracellular matrix (ECM)-receptor interactions were markedly enriched, while Masson's trichrome staining revealed decreased collagen content in LW. IF staining showed increased COL6A3 expression and decreased FN1 expression in LW tissues. Taken together, our study provides a descriptive single-cell atlas of late-stage wound tissues with differing regenerative outcomes, indicating differences in keratinocyte composition, transcriptional states, and intercellular communication. Collectively, our findings offer a resource for understanding cellular heterogeneity associated with WIHN and may inform future studies aimed at elucidating the mechanisms of hair follicle regeneration in cutaneous repair.

Biological systems rely on asynchronous and temporally overlapping dynamics, allowing for the concurrent activation of multiple processes. This principle is particularly evident in brain function, where cognitive tasks engage distributed, interacting regions rather than sequentially isolated ones. To investigate the mechanisms enabling such coordination, we study a modular spiking neural network composed of leaky integrate-and-fire neurons and governed by spike-timing-dependent plasticity. Our model stores modular spatiotemporal patterns both at the mesoscopic level (sequences of modules) and at the microscopic level (precise spike timings) and includes a parameter, , which regulates the degree of temporal overlap between modules' activations. By tuning , the network transitions from sequential to overlapping regimes, ranging from synfire chainlike dynamics to fully co-activated modules. We investigate how the temporal structure influences the network's capacity to encode and selectively retrieve multiple dynamical patterns while considering biological constraints such as the cost of long-range connectivity. Our results offer insight into how spatiotemporal coding and network organization support robust, large-scale memory storage and replay.

The brain criticality hypothesis has largely only characterized brain dynamics in terms of their self-similarity, although experimental evidence suggests that the brain exhibits significant multifractality. To understand how multifractality may emerge in critical-like systems modeling neuronal activity, we used a neural field model exhibiting neural oscillations and a critical phase transition. We find that multifractality emerges near a synchronization phase transition, and that the pattern of variation in multifractality changes when placing the model at a different phase transition. These findings show that multifractality in temporal dynamics emerges near criticality in neural fields, providing a formal basis for interpreting multifractality in brain recordings.

Inhibitory interneurons in the cortex are classified into cell types differing in their morphology, electrophysiology, and connectivity. Although it is known that parvalbumin (PV), somatostatin (SST), and vasoactive intestinal polypeptide-expressing neurons (VIP), the major inhibitory neuron subtypes in the cortex, have distinct modulatory effects on excitatory neurons, how heterogeneous spatial connectivity properties relate to network computations is not well understood. Here, we study the implications of heterogeneous inhibitory neurons on the dynamics and computations of spatially structured neural networks. We develop a mean-field model of the system in order to systematically examine excitation-inhibition balance, dynamical stability, and cell-type specific gain modulations. The model incorporates three inhibitory cell types and excitatory neurons with distinct connectivity probabilities and recent evidence of long-range spatial projections of SST neurons. Position-dependent firing-rate predictions are validated against simulations, and balanced solutions under Gaussian assumptions are derived from scaling arguments. Stability analysis shows that while long-range inhibitory projections in E-I circuits with a homogeneous inhibitory population result in instability, the heterogeneous network maintains stability with long-range SST projections. This suggests that a mixture of short- and long-range inhibitions may be key to providing diverse computations while maintaining stability. We further find that conductance-based synaptic transmissions are necessary to reproduce experimentally observed cell-type-specific gain modulations of inhibition by PV and SST neurons. These gain modulations are distance dependent, and a linear response analysis suggested that shifts in the excitation-inhibition balance of the network underlie these effects. Our theoretical approach offers insight into the computational function of cell-type-specific and distance-dependent network structure.