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Multidrug-resistant (MDR) Escherichia coli represents a larger fraction of the E. coli population in preweaned dairy calf feces compared to older postweaned calves and adult cows. Previous work demonstrated that the MDR genotype is strongly associated with iron-acquisition genes (iucABCD-iutA and sitABCD), which are primarily carried on IncFIB plasmids that frequently co-encode these systems with antimicrobial-resistance genes in bovine E. coli. We hypothesized that iron limitation in the preweaned calf gut favors E. coli carrying these iron-acquisition systems, thereby indirectly selecting for antimicrobial resistance. To test this, we quantified growth by isothermal microcalorimetry in bovine cecal medium and used RT-qPCR to measure expression of iron-acquisition genes under iron-replete and iron-limited conditions. Under iron-replete conditions, plasmid carriage imposed a negligible fitness cost. Under iron limitation, IncFIB-positive strains generated up to 3.5-fold more cumulative heat and sustained metabolic activity 6-10 h longer than their plasmid-negative counterparts. This advantage corresponded to strong siderophore gene upregulation, with iucA expression increased by up to 38.4-fold. Multivariate analysis further distinguished plasmid-bearing from plasmid-free groups under iron stress. The results of this study demonstrate that plasmid maintenance costs are negligible in rich medium, and they are fully offset by the significant growth advantage conferred by iron-acquisition systems in iron-limited growth conditions. These findings suggest that gut iron availability contributes to the maintenance of antimicrobial resistance in young calves and highlight nutrition-based strategies as potential interventions to reduce antimicrobial resistance in these animals.IMPORTANCEThe preweaned calf gut is an iron-poor environment that favors Escherichia coli carrying IncFIB plasmids encoding both siderophore systems and antimicrobial resistance genes. Our findings show that these plasmids convert a potential liability, the cost of extra DNA, into a fitness advantage under iron limitation. This nutritional-genetic interaction explains why multidrug-resistant strains persist in young calves and suggests that dietary iron management could help reduce antimicrobial resistance in livestock, with implications for food safety and public health.

Multidrug-resistant Klebsiella pneumoniae is a global health threat circulating across humans, animals, food, and environments. Despite increasing One Health surveillance, it remains unclear whether the dominant antimicrobial resistance (AMR) reservoirs in K. pneumoniae are universal or depend on regional ecological context. Here, a synchronized One Health survey in Pingguo, China (5384 samples) is integrated with parallel cross-niche datasets (Italy, Ghana) and a 69184 public genome collection, mapping AMR ecology via network metrics and lineage-aware mixed models. In Pingguo, high-risk determinants are enriched in humans, whereas overall AMR burdens and multidrug resistance platforms are enriched outside humans. Across the public dataset, AMR reservoir dominance varies eco-geographically, with nonhuman isolates carrying higher AMR gene burdens in the China subset (mean +4.5 per isolate), largely due to animal and environmental isolates, while human isolates carry higher burdens in the U.S. and Europe subsets. Lineage explains 44.8% of the burden difference. Remaining within-lineage differences, reservoir-matched integron enrichment, and region-specific connectivity suggest additional horizontal gene transfer-related contributions. This is supported in Pingguo by a highly structured cross-niche plasmid-sharing network shaped by specific plasmid groups and successful lineages. Overall, dominant AMR reservoirs are eco-geographically contingent. Integron load and cross-niche connectivity provide indicators for tiered, region-tailored surveillance.

Abdominal aortic aneurysm (AAA) is a progressive vascular disease associated with high morbidity and mortality. Translational imaging and interventional research are limited by the availability of standardized large-animal models. This exploratory study compared different endovascular and pharmacological strategies for AAA induction in swine and characterized associated morphological and inflammatory vessel wall changes. 19 female German Landrace swine were enrolled, with 18 included in the final analysis (n = 3 per group). Animals were assigned to five intervention groups and one untreated control group. Interventions comprised combinations of mechanical aortic dilation, intra-aortic administration of collagenase (6,000 IU), elastase (500 IU), calcium chloride (CaCl2, 25%; group-specific dosing), and oral β-aminopropionitrile (BAPN; 0.05-0.15 mg/kg). Aortic diameter changes and molecular markers of inflammation and vascular remodeling were assessed. Given the limited group size, analyses were considered exploratory and hypothesis-generating. Mechanical dilation combined with enzymatic and CaCl2 exposure resulted in the largest mean increase in aortic diameter (1.63 ± 0.25 cm vs. 0.61 ± 0.04 cm in controls). This group was associated with increased Galectin-3 expression (89.69 ± 7.02%). BAPN alone was associated with smaller aortic dilations (0.73 ± 0.06 cm) and marked reduction of vascular smooth muscle cell markers, including α-smooth muscle actin (23.50 ± 4.82%), consistent with active extracellular matrix remodeling. Within this exploratory large-animal setting, combined mechanical and enzymatic injury was associated with more pronounced structural remodeling compared to isolated approaches, whereas BAPN predominantly induces early vessel wall degeneration. The graded phenotypes observed across groups provide a comparative framework for future preclinical imaging-based and interventional AAA research. · The porcine models enable comparative preclinical investigation of graded abdominal aortic aneurysm phenotypes.. · Combined mechanical dilation and enzymatic injury is associated with more pronounced structural remodeling in swine.. · Greater structural remodeling is accompanied by increased macrophage-associated inflammatory markers.. · Vascular smooth muscle cell markers are reduced in association with vessel wall degeneration.. · Ranner-Hafferl M-LHH, Mangarova DB, Heyl JL et al. Characterization of Aortic Degeneration and Inflammatory Profiles in Porcine Models of Abdominal Aortic Aneurysm. Rofo 2026; 10.1055/a-2854-8724. Das abdominelle Aortenaneurysma (AAA) ist eine progrediente Gefäßerkrankung mit hoher Morbidität und Mortalität, bei der die translationale Forschung aufgrund eines Mangels an etablierten Großtiermodellen eingeschränkt ist. In dieser explorativen Studie wurden verschiedene endovaskuläre und pharmakologische Strategien zur AAA-Induktion im Schweinemodell vergleichend untersucht und die damit assoziierten morphologischen sowie inflammatorischen Veränderungen der Gefäßwand charakterisiert.Insgesamt wurden 19 weibliche Deutsche Landrasse-Schweine für die Studie verwendet, davon wurden 18 in die finale Analyse eingeschlossen. Die Tiere wurden fünf Interventionsgruppen sowie einer unbehandelten Kontrollgruppe zugeteilt (n = 3 pro Gruppe). Die Interventionen umfassten Kombinationen aus mechanischer Aortendilatation, intraaortaler Applikation von Kollagenase (6.000 IE), Elastase (500 IE), Calciumchlorid (CaCl2, 25%; gruppenspezifisch) sowie oraler Gabe von β-Aminopropionitril (BAPN; 0,05–0,15 mg/kg). Analysiert wurden Aortendurchmesser sowie molekulare Marker der Inflammation und des Gefäßumbaus. Aufgrund der begrenzten Gruppengröße wurden die Analysen als explorativ und hypothesengenerierend eingestuft.Die Kombination aus mechanischer Dilatation und enzymatischer sowie CaCl2-induzierter Schädigung war mit der größten mittleren Zunahme des Aortendurchmessers assoziiert (1,63 ± 0,25 cm vs. 0,61 ± 0,04 cm in der Kontrollgruppe). Diese Gruppe zeigte zudem eine erhöhte Galectin-3-Expression (89,69 ± 7,02%). Die alleinige BAPN-Gabe war mit geringeren Durchmesserzunahmen assoziiert (0,73 ± 0,06 cm) und ging mit einer deutlichen Reduktion glatter Muskelzellmarker, einschließlich α-Smooth-Muscle-Actin (23,50 ± 4,82%), einher, vereinbar mit aktivem Umbau der extrazellulären Matrix.In diesem explorativen Großtiermodell war die kombinierte mechanisch-enzymatische Gefäßschädigung mit ausgeprägterem strukturellem Umbau im Vergleich zu isolierten Ansätzen assoziiert, während BAPN überwiegend frühe Stadien der Gefäßwanddegeneration induzierte. Die beobachteten abgestuften Phänotypen bieten einen vergleichenden Rahmen für zukünftige präklinische Untersuchungen im Bereich bildgebender und interventioneller AAA-Forschung. · Die porzinen Modelle ermöglichen eine vergleichende präklinische Untersuchung abgestufter Phänotypen des abdominellen Aortenaneurysmas.. · Die kombinierte mechanische Dilatation und enzymatische Schädigung ist mit ausgeprägterem strukturellem Umbau assoziiert.. · Stärkeres Remodeling geht mit einer erhöhten Expression makrophagenassoziierter inflammatorischer Marker einher.. · Marker glatter Gefäßmuskelzellen sind im Zusammenhang mit Gefäßwanddegeneration vermindert nachweisbar..

Numerous studies indicate that smoking during pregnancy has harmful effects on the offspring. Prenatal smoke exposure (PSE) may lead to fetal hypoxia and ischemia, which negatively affect brain development and increase the risk of neurological deficits. However, its long‑term impact on retinal vulnerability in adulthood is less well understood. To investigate the effects of prenatal smoke exposure on retinal structure and vulnerability in a chronic retinal hypoperfusion model in adult rats. Wistar rats were mated and exposed to whole‑body tobacco smoke for 2 hours daily from mating until delivery, using a closed‑chamber manual smoking system with four research cigarettes per occasion, modelling passive smoking. Neurobehavioral development was assessed in newborn rats during the first weeks of life. At 5 months of age, permanent bilateral common carotid artery occlusion (BCCAO) was performed under isoflurane anaesthesia via midline neck incision. Two weeks after BCCAO, all animals were sacrificed with an overdose of anaesthetic, and eyes were processed for histological analysis. Retinal layer thickness (outer and inner nuclear and plexiform layers) and cell counts per 100 µm in the ganglion cell layer (GCL) were measured. BCCAO resulted in markedly reduced retinal layer thickness and morphological signs of degeneration with individual variation in all layers compared to sham‑operated controls. The number of cells in the GCL decreased by approximately 50%. Prenatal smoke exposure alone also led to a significant reduction in GCL cell number. While our previous work had shown only minor retardation of neurobehavioral development in prenatally smoke‑exposed neonatal rats, the present study demonstrated pronounced histological damage in the retina of adult rats subjected to PSE, with further exacerbation after chronic hypoperfusion. Prenatal exposure to tobacco smoke induces long‑lasting structural alterations in the retina and increases susceptibility to later hypoperfusion‑induced retinal injury in adult rats. These findings support the concept that adverse intrauterine exposures have persistent consequences for neural tissues and underscore the importance of avoiding smoking during pregnancy.

Charles Darwin was profoundly fascinated by Drosera (sundew plants), which have evolved as flesh-eaters with modified leaves, called traps, to catch small animals yet do not respond to transient mechanical stimuli. Despite Darwin's early insights, and recent advances in characterizing these mechanisms, our knowledge of the signaling molecules underlying prey capture responses remains limited. Here, we identify glutathione (GSH) as an important mediator within the signaling cascade of sundew carnivory, implicated in trap closure in a calcium-dependent manner. We show that prey capture promotes GSH accumulation in sundew leaves, while GSH depletion inhibits the movement. Furthermore, application of GSH is sufficient to induce trap closure across the Droseraceae family, including both the snap traps of Venus flytrap and the adhesive traps of sundew species. These findings advance our understanding of the enigmatic mechanisms of plant carnivory, a phenomenon that has intrigued scientists since Darwin's era.

Wings are a key trait innovation in the evolutionary history of insects, and contributes to the largest diversity of animals on the planet. However, we still have an incomplete understanding of the functional changes in genes behind this diversification. Insect, Malacostraca and Chelicerata species originated as primitive arthropods during the Cambrian period. Malacostraca as the ancestral taxa of winged insects, are key to understanding this radiation. Here, a deep learning (DL) model for wing genes identification (DeepWG) based on bidirectional long short-term memory (BiLSTM) and attention mechanism (AM) was constructed based on the protein sequences of 119 species. DeepWG demonstrated a strong potential in mining key genes of insect wings, achieving an accuracy rate of 97.3% on the test set. Our research found that the 351 key genes identified by DeepWG and their orthologs exhibit transcriptional similarity in wing and gill tissues, providing molecular evidence consistent with the Ttracheal gill theory of insect wing origin. This study not only proposes a new method for identifying key genes, but also lays the foundation for genetic studies of key evolutionary adaptations in winged insects.

Ocean acidification is known to affect calcified structures in marine organisms, yet its impact on non-calcified but functionally essential feeding tools remains poorly understood. The radula is a defining molluscan apomorphy, whose mechanical performance is critical for feeding and survival. Here we investigated the effects of reduced seawater pH on the radular teeth of the intertidal gastropod Littorina littorea. Individuals were maintained for seven weeks under acidified conditions (pH 7.5) or near-present-day conditions (pH 8.1) and compared with a field-collected control group. Radulae were analysed using scanning electron microscopy, confocal laser scanning microscopy, energy-dispersive X-ray spectroscopy, and nanoindentation. Radulae from acid-treated individuals exhibited markedly increased tooth wear in the working zone despite largely preserved gross morphology. Wear was most pronounced at the cusps of central and lateral teeth and showed rounded profiles indicative of progressive abrasive wear. Acidic conditions caused pronounced changes in the outer tooth coating, including reduced silicon enrichment and substantial decreases in stiffness and hardness, while the inner tooth structure was only weakly affected. Confocal microscopy revealed treatment-specific autofluorescence patterns, suggesting pH-dependent alterations of the organic matrix. Differences between laboratory-maintained and field-collected individuals further indicate that feeding conditions influence radular tooth properties. These results demonstrate that ocean acidification can impair radular function through material-level degradation of composite feeding structures, potentially reducing grazing efficiency and imposing sublethal fitness costs.

Many animals efficiently interpret their environment by detecting geometric features like corners, highlighting the power of feature extraction for reducing visual complexity; similarly, with the surge in visual data, nature-inspired optical corner detection offers a promising yet still elusive solution for energy-efficient information processing and compression. Here, we propose a universal strategy for optical corner imaging with azimuthal Hilbert transformation metasurfaces. Multiple objects, regardless of their amplitude, phase, or angular characteristics, can be detected simultaneously with a single metasurface, featuring broadband and full-field-of-view properties. Trade-offs between spatial and angular resolution are assessed, offering practical guidance for implementation. We further demonstrate motion tracking as a proof-of-concept application leveraging the data-compressed corner imaging framework. This work paves the way for next-generation optical information processing technologies.

Genome-wide association studies (GWAS) have provided strong evidence that modifiers of CAG tract length have a crucial influence on Huntington disease onset, but somatic expansion alone may not be sufficient to drive neuronal death. Here, we report that DSBs drive neuropathology in male HdhQ(150/150) mice, regardless of somatic expansion of the inherited disease allele. DSBs and somatic expansion occur simultaneously in the HD brain, but the two types of DNA damage drive disease by distinct mechanisms. The site-specific increases in CAG tract length are driven by active mismatch repair (MMR), while DSBs occur genome-wide and are driven by mutant huntingtin-mediated suppression of nonhomologous joining of DNA broken ends. DSBs and transcriptional dysfunction occur in animals that cannot somatically expand their inherited allele. Conversely, suppression of DSBs is sufficient to reverse neuropathology even when somatic expansion is active. We propose that CAG expansion and DSBs promote downstream neuronal pathology as separable drivers. The disease-length CAG tract leads to early inhibition of DSBR and accumulating DSBs over time ultimately kill neurons.

Prefrontal neurons exhibit diverse activity during cognitive functions such as working memory, attention, and timing; however, the importance of this heterogeneity is unclear. Our goal was to better understand the diversity of prefrontal activity through anatomical connectivity. We harnessed circuit-specific tools in mice to capture activity within prefrontal projections during interval timing, a highly translational cognitive process that requires working memory for temporal rules and attention to the passage of time to estimate a temporal interval of several seconds. We used neuronal recordings to capture prefrontal activity during interval timing, with major patterns characterized by monotonic time-dependent ramping over a temporal interval. We then leveraged retrograde viruses to interrogate prefrontal cortex (PFC) projections to the mediodorsal thalamus (PFC-MD) and the dorsomedial striatum (PFC-DMS). We report three main findings. First, circuit-specific fiber photometry revealed that PFC-MD and PFC-DMS activity encoded distinct temporal signals, with PFC-MD projections ramping down and PFC-DMS ramping up to interval timing response times. Second, circuit-specific inactivation revealed that suppressing PFC-DMS projections disrupted animals' internal estimates of time. Third, circuit-specific single-nucleus RNA sequencing of projection-defined prefrontal neurons revealed distinct transcriptomic profiles of PFC-MD and PFC-DMS projections, with enrichment of cortical layer-associated genes as well as genes such as Cux2, Camk2n1, Htr4, and Foxp2. These data suggest that differences in gene expression and connectivity distinguish prefrontal activity during interval timing. These findings advance our fundamental understanding of prefrontal function and dysfunction in human disease.

Humans rely heavily on visual function to gather information, and loss of vision has a significant impact on quality of life. However, it is difficult to quantitatively evaluate the effects of chemical compounds on visual function in toxicity studies using animals (such as OECD guideline studies). Consequently, evaluation including ophthalmological and histopathological examinations has played a major role to date. Visually evoked potential (VEP) is a type of brain wave that reflects the activity of the entire visual pathway, including the retina, optic nerve, and visual cortex. We investigated whether VEP could detect the effects of acrylamide, a toxicant known to affect peripheral nerves, on visual function. Acrylamide was administered to rats via drinking water at concentrations of 0 (control) and 200 ppm for 4 weeks, and electroretinograms (ERGs) and VEPs were recorded at weeks 0, 2, and 4. After the 4-week treatment period, the eyes and optic nerves were examined by light microscope. Acrylamide exposure significantly delayed VEP latency, while no effects were observed on the retina and optic nerve by ERG or histopathology. A significant decrease in grip strength in the hindlimbs and degeneration of sciatic nerve fibers were observed in the acrylamide-treated group, indicating that acrylamide damaged peripheral nerves. In conclusion, our study demonstrated that VEP can detect the effects of acrylamide on visual function earlier than histopathological examination, suggesting that VEP could be useful for detecting early-phase effects of chemical compounds on visual function and for evaluating whether morphological changes observed in toxicity studies are toxicologically significant.

The human gut microbiome is shaped by diverse selective forces that originate from host and environmental factors and it substantially influences health and disease. Whereas the association of microbial lineages with various health conditions has been shown at different taxonomic levels1-5, the extent to which unifying adaptive mechanisms sort microbial lineages into ecologically differentiated populations remains poorly understood. Here we show that genome-wide selective sweeps are a pervasive mechanism that differentiates bacteria in the microbiome. This mechanism leads to population structures akin to global epidemics across geographically and ethnically diverse human populations. Such sweeps arise when an adaptation allows a clone to outcompete others in its niche followed by rediversification, and they manifest as clusters of closely related genomes on long branches in phylogenetic trees. This structure is revealed by excluding recombination events that mask the clonal descent of the genomes. Indeed, we show that genome-wide sweeps originate under a wide range of recombination rates in at least 66 taxa from 25 bacterial families. Estimated ages of divergence suggest that sweep clusters can spread globally within decades and that this process has occurred throughout human history. Sweep clusters are associated with different host conditions-such as age, colorectal cancer, inflammatory bowel diseases and type 2 diabetes-as an indication of their ecological differentiation. Our results reveal an evolutionary mechanism for the observation of stably inherited strains with differential associations and provide a theoretical foundation for analysing adaptation among microbial populations.

Early intervention in metabolic dysfunction-associated steatohepatitis (MASH) is critical to halt disease progression. However, noninvasive tools for monitoring early-stage MASH and therapeutic efficacy in preclinical models remain limited, impeding preclinical drug development. This study establishes an integrated approach using multiparametric magnetic resonance imaging (MRI) at 9.4&#x2009;T to dynamically track disease development and drug response in a prefibrotic MASH mouse model. Mice were fed a high-fat and high-cholesterol diet (HFHCD) for 16 weeks to induce early MASH without fibrosis and treated with the anti-MASH drug semaglutide for 8 weeks after modeling. Longitudinal MRI assessments-including proton density fat fraction (PDFF), 1H magnetic resonance spectroscopy (MRS), T2 mapping, and diffusion-weighted imaging (DWI)-were performed every 4 weeks and correlated with histopathology. Histology confirmed early MASH after 16 weeks, without fibrosis. All MRI parameters strongly correlated with histopathological scores. HFHCD feeding led to significant changes: PDFF, MRS-derived liver fat content (LFC), and T2 values increased by 6.8-, 5.2-, and 2.5-fold, respectively, while apparent diffusion coefficient (ADC) decreased by 30% (p&#x2009;<&#x2009;0.001). T2 and ADC also correlated with MRS-quantified saturated fatty acids. Semaglutide treatment effectively reversed these changes: PDFF decreased by 73%, LFC by 62%, T2 by 46% (p&#x2009;<&#x2009;0.001), and ADC increased 1.4-fold (p&#x2009;=&#x2009;0.017) compared to the vehicle group. This work demonstrates multiparametric MRI as a powerful noninvasive platform for monitoring early MASH dynamics and treatment response. By enabling longitudinal assessment in a prefibrotic model, this approach accelerates translational research in MASH diagnosis and drug development. The established multiparametric MRI evaluation system provides a valuable noninvasive monitoring platform for preclinical early-stage MASH research, demonstrating significant potential to accelerate the translational progress in MASH diagnosis and drug development. A novel prefibrotic MASH model was established to assess early-stage MASH progression. Multiparametric MRI at 9.4&#x2009;T enables noninvasive, longitudinal monitoring of early MASH. Semaglutide-induced improvement in steatosis and inflammation can be monitored by multiparametric MRI.

Antimicrobial resistance (AMR) in common bacterial pathogens, including methicillin-resistant Staphylococcus aureus (MRSA), is an increasingly dire public health threat, with MRSA accounting for up to 90% of S. aureus infections. To expand the treatment arsenal against MRSA infections, we developed a class of tunable three-dimensional tricyclic 2-pyridones, termed TriPcides, that can kill MRSA resistant to last-resort antibiotics and eliminate MRSA persister cells. No preexisting resistance was detected across hundreds of clinical isolates, and continuous exposure of MRSA to TriPcides did not elicit detectable resistance. Treatment with TriPcides causes a rapid decrease in membrane integrity and increased levels of reactive oxygen species. Last, TriPcides effectively reduce secretion of important virulence factors and result in reduced ulcer size and healing time in S. aureus murine skin and soft tissue infections but do not reduce bacterial burden.

To determine whether high-dose sucralose (SUC) exposure alters circadian organization of the mouse extraorbital lacrimal gland, immunometabolic pathways, and tear film function using multi-omics and functional assays. Male C57BL/6J mice received SUC (0.72&#xa0;mg/mL in drinking water) or water under a 12:12 hour light-dark cycle for 30 days. Extraorbital lacrimal glands were collected every 3 hours across 24 hours (n = 3/time point/group) for RNA sequencing and 4D-data-independent acquisition proteomics. Rhythms were identified with JTK_CYCLE, differential expression with DESeq2/MSstats followed by Kyoto Encyclopedia of Genes and Genomes pathway analysis. Oil Red O and CD4/CD8 immunohistochemistry assessed lipid and immune signatures. Tear secretion, tear film breakup time, and corneal fluorescein staining were measured at ZT0, ZT6, ZT12, and ZT18. SUC increased rhythmic transcripts (3074 to 4600) and proteins (378 to 984), redistributing acrophases (transcript peaks shifted toward ZT3-ZT6; protein peaks toward ZT21-ZT24), indicating clock phase remodeling. Both omics layers converged at the pathway level, showing downshifts in metabolic and immune programs, including complement- and T-cell-related signatures. SUC elevated water intake and body weight without altering glucose tolerance. Functionally, SUC-treated mice showed reduced stimulated tear secretion, whereas tear film breakup time and corneal fluorescein staining did not differ significantly. Under this high-exposure paradigm, SUC was associated with lacrimal circadian remodeling, reduced metabolic and immune pathway signatures, and lower stimulated tear secretion, identifying candidate pathways for mechanistic testing without establishing direct causality.

Birds and mammals exhibit extraordinary facial diversity, reflecting adaptations to distinct ecological niches and feeding strategies. While core face-building developmental programs are conserved and orchestrated by interactions between ectodermal organizers and the underlying mesenchyme, mechanisms driving facial shape variation remain poorly understood. Here, we integrate single-cell transcriptomic and chromatin accessibility profiling of mouse and chicken developing face to construct a comparative regulatory map. Although both ectodermal and mesenchymal populations display distinct regulatory features in each species, the mesenchyme exhibits markedly greater divergence, pointing to its central role in shaping facial morphology. We further reveal unexpected molecular complexity in the main face-shaping organizer, including a mouse-specific Shh/Wnt5a expression domain. At key morphogen loci (Bmp4, Fgf8, and Wnt5a), conserved and lineage-specific enhancers exhibit spatially restricted activity patterns that mirror divergent signaling domains. These findings demonstrate how cis-regulatory evolution modulates conserved developmental programs to generate morphological novelty, providing a valuable resource for studying vertebrate facial evolution.

Luteolin (LUT) is a biologically active flavonoid exhibiting significant therapeutic potential against neurodegenerative disorders such as cognitive impairment. Nonetheless, its clinical application is limited by poor aqueous solubility, extensive first-pass metabolism and diminished permeability across the blood-brain barrier (BBB) and consequently, low oral bioavailability. This study aimed to develop PEGylated LUT nano-vesicular delivery systems to enhance brain delivery and therapeutic efficacy. PEGylated LUT-Aspasomes were prepared via thin-film hydration, using Brij 52 for PEGylation. Sixteen formulations were designed using a 24 full factorial design, varying the weights of Ascorbyl Palmitate (AP), Cholesterol (CH), Brij 52, and Span (Sp) type. The optimized formulation (F14) is composed of 40 mg AP and CH, 10 mg Brij 52, and 50 mg Sp 60 with a desirability value (0.747). F14 demonstrated entrapment efficiency (EE%) (86.24&#x2009;&#xb1;&#x2009;1.16%), particle size (PS) (188.30&#x2009;&#xb1;&#x2009;1.50 nm), and zeta potential (ZP) (-25.31&#x2009;&#xb1;&#x2009;0.84 mV). TEM confirmed spherical, nanosized vesicles with uniform morphology. XRD analysis revealed transformation of LUT from crystalline to amorphous form, supporting successful encapsulation. Drug release and ex-vivo nasal permeation were enhanced 3.4- and fourfold, respectively, compared to free LUT. In-vivo studies in chronic unpredictable stress (CUS) rats revealed that the optimized PEGylated LUT-Aspasomes significantly enhanced neuroprotective effects compared to free LUT. Treated rats exhibited improved spatial memory and a 1.7-fold greater attenuation of depressive-like behavior relative to free LUT. Biochemically, F14 produced 1.15-, 1.23-, and 1.06-fold superior reductions in hippocampal acetylcholinesterase (AChE) activity, serum corticosterone (CORT), and increased brain-derived neurotrophic factor (BDNF) levels, respectively, compared to LUT suspension. Histopathological examination confirmed nearly preserved hippocampal architecture in PEGylated LUT-Aspasome-treated rats. These effects are attributed to LUT encapsulation within PEGylated Aspasomes, which enhanced stability, brain delivery, and neuroprotective efficacy. Thus, PEGylated LUT-Aspasomes are a promising strategy for managing stress-related neurobehavioral disorders.

MicroRNA (miRNA), a class of short non-coding RNA, serves as a metabolically cheap and fast acting mechanism to inhibit translation of complementary mRNA. These molecules play a central role in regulating metabolic changes during torpor in small mammals, such as the house mouse (Mus musculus). The present study sequenced mouse kidney RNA to identify changes in miRNA expression in kidney, comparing active control mice (C57BL/6 strain) with mice in varying stages of torpor. Out of a total 962 miRNAs, 82 were differentially expressed when comparing torpid states to the control with 65, 14, and 2 miRNAs differentially expressed in deep torpor, early torpor, and recovery from torpor mice, respectively. In silico analysis of these targets demonstrated a clear downregulation of gene sets associated with cancer and active metabolism. This suggests that miRNA may play important roles in suppressing cell division and proliferation during torpor, along with maintaining a hypometabolic state. Machine learning techniques identified two specific miRNAs (miR-101c, miR-101a-3p) that can strongly distinguish between torpid and active states in kidney among other differentially regulated miRNA. The present study highlights the role of miRNA in transcriptional regulation of cell signaling and metabolic pathways in kidney throughout mouse torpor.

T cell exhaustion is a major barrier to effective antitumor immunity, yet the tumor-intrinsic mechanisms remain poorly defined. Through single-cell and spatial proteomics analyses of esophageal squamous cell carcinoma (ESCC), we uncover two infection-like CD8+ T cell trajectories, acute-like and chronic-like responses, whose fates are dictated by the tumor cell subtypes they encounter. This concept links tumor heterogeneity to the shaping of local immune niches. Mechanistically, we identify CDC28 protein kinase regulatory subunit 1B (CKS1B) as a tumor-intrinsic inducer of chronic-like exhaustion. CKS1B forms a complex with S-phase kinase-associated protein to promote interferon regulatory factor 3 (IRF3) ubiquitination and degradation, thereby suppressing type I interferon signaling and antigen presentation. This impairs tumor cell elimination and drives progressive CD8+ T cell stimulation and exhaustion. Pharmacological blockade of the CKS1B-IRF3 interaction with 14i restores CD8+ T cell function and synergizes with immune checkpoint blockade. The tumor-intrinsic oncogenic-immune axis, which connects cancer cell signaling to immune dysfunction, is conserved across multiple malignancies, establishing a conceptual and therapeutic framework for overcoming tumor-driven T cell exhaustion.