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Dengue virus (DENV) infection poses a major global public health burden, particularly in endemic regions where repeated exposure increases the risk of severe disease. Despite revisions to the World Health Organization (WHO) dengue classification, early prediction of progression to severe dengue remains challenging due to overlapping clinical and laboratory features. Current management strategies rely primarily on supportive care and reactive monitoring, underscoring the need for predictive biomarkers that enable early risk stratification and timely intervention. COMBAT is a prospective, multicenter, observational longitudinal study conducted in dengue-endemic regions of Guatemala and India. Patients will be classified according to WHO 2009 criteria into dengue without warning signs, dengue with warning signs, and severe dengue, alongside age- and sex-matched healthy controls. A single blood sample will be collected from non-hospitalized patients while two blood samples per participant will be collected during hospitalization and one at discharge. Multi-omics analyses, including transcriptomics, proteomics, glycomics and metabolomics, will be performed in a discovery cohort and validated in an independent cohort. Integrated systems biology approaches will be used to identify host immune and metabolic pathways associated with dengue severity in a mechanism-based prognostic biomarker discovery. This study aims to generate comprehensive systems-level insights into host-virus interactions driving dengue severity and to identify biomarkers predictive of disease progression. The findings may inform improved patient triage, early intervention strategies, and the identification of novel therapeutic targets. ClinicalTrials.gov ID NCT06751836 Registration Date: December 13, 2024, CTRI/2022/10/046293 (MAHE) Registration Date: October 10, 2022.

Spinal cord injury (SCI) precipitates a multiphasic secondary injury cascade that establishes a hostile, inhibitory microenvironment, rendering the condition refractory to conventional surgical stabilization and rehabilitation. While cell-based therapies offer promise for neural reconstruction, their clinical translation is impeded by protocol heterogeneity and fragmented safety data. To address this, we mapped clinical trials for SCI to quantify patient demographic parameters, identify lineage-specific adverse-event patterns, evaluate objective motor and sensory efficacy outcomes, and analyze methodological trial designs to formulate concrete structural recommendations for future advanced-phase trials. We analyzed clinical trials from the Web of Science Core Collection (SCIE and ESCI) published since 2005. The dataset comprised 116 eligible studies involving patients with SCI receiving cellular therapies with reported safety outcomes. We employed a dual-method approach combining manual extraction of clinical characteristics (demographics, interventions, adverse events, efficacy outcomes) with quantitative data analysis. Statistical associations between therapeutic variables (cell type, route, dosage) and safety profiles were evaluated using Fisher's exact test. The clinical landscape is predominantly defined by early-phase (Phase 1: 59.5%), single-arm investigations (63.8%) utilizing autologous bone marrow-derived cells. Reflecting a cautious paradigm to minimize severe complications, patient selection frequently targeted the hemodynamically stable chronic phase (58.3%) and thoracic SCI (21.6%). Safety analyses revealed lineage-specific profiles: mesenchymal stromal cells were significantly associated with transient fever (P &#x2009;=&#x2009;0.038), whereas intrathecal administration correlated with procedural symptoms such as headache (P <&#x2009;0.001). The observation of higher systemic adverse event rates in low-dose cohorts was likely confounded by the mandatory concurrent immunosuppressive regimens required for specific allogeneic lineages, rather than the absolute cell dose. Regarding therapeutic efficacy, outcomes were critically influenced by the chronological phase of injury. Patients treated in the acute or subacute phases exhibited higher rates of neurological improvement, though distinguishing this from spontaneous recovery remains challenging, whereas chronic phase interventions demonstrated more limited primary sensorimotor gains. Additionally, intrathecal administration showed an advantage in preserving sensory pathways due to minimized structural disruption, while dose requirements could not be generalized and varied fundamentally based on specific cellular mechanisms of action. While the baseline safety of cellular transplantation for spinal cord injury is established, clinical translation remains hindered by methodological heterogeneity, imprecise patient stratification, and a reliance on single-arm trial designs. To navigate this translational bottleneck, future investigations should adopt multi-tiered methodological frameworks. First, study designs should transition toward controlled protocols, utilizing crossover designs for chronic cohorts and matched historical or synthetic controls for acute and subacute phases. Concurrently, patient selection must evolve from broad clinical grading to advanced stratification. Integrating biomarkers, electrophysiology, and imaging to objectively quantify tissue sparing can better identify responsive subgroups, thereby improving trial efficiency and accelerating clinical translation. Beyond cohort refinement, intervention parameters, specifically dosage and delivery routes, should be individualized according to cell lineage, as adverse events associate more with intrinsic cellular biology and procedural invasiveness. Furthermore, isolating the true therapeutic effect requires the standardization and reporting of confounding variables, such as immunosuppressive regimens and physical rehabilitation. Finally, by separating shorter-term efficacy measurements from long-term safety registries, the field can facilitate a more reliable and objective clinical translation.

This critical narrative review evaluates saliva, sweat, and exhaled breath as noninvasive interface matrices for exercise chemistry rather than as general substitutes for blood or urine. It synthesizes peer-reviewed evidence on exercise-related sampling biology, analytical platforms, quantitative performance, validation limits, and translational readiness; because protocols and platforms remain heterogeneous, the evidence is interpreted qualitatively rather than pooled meta-analytically. Saliva is strongest for repeated neuroendocrine, autonomic, immune, oxidative, and short-latency metabolite measurements, but its interpretation depends on flow rate, timing, oral contamination, and normalization. Sweat is most mature for sweat rate and sodium/chloride loss monitoring and is uniquely compatible with wearable platforms, whereas sweat lactate, glucose, cortisol, and cytokines require stronger physiological validation before they can be treated as systemic markers. Exhaled breath offers second-scale access to volatile metabolism, substrate use, and airway responses, but breathomics remains instrumentally demanding and vulnerable to ventilation, diet, humidity, and ambient-air confounding. The review concludes that useful applications should match each analyte and platform to a defined exercise question, a matrix-specific validation model, and a practical decision context.

Accurate prediction of cancer drug responses is essential for advancing cancer treatment strategies and drug development. With the increasing availability of large-scale pharmacogenomic datasets, many deep learning models have been proposed to predict cancer drug responses. However, many existing models lack the capacity to offer critical biomedical insights, such as providing interpretability regarding the potential mechanism of action. We propose DR.DEGMON (Drug Response prediction using Differentially Expressed Genes with Multi-layer perceptron integrating gene Ontology Network), a self-explainable deep neural network designed to predict the viability of pan-cancer cell lines in response to drug treatments by utilizing differentially expressed genes. DR.DEGMON leverages prior biological knowledge by incorporating Gene Ontology (GO) into the hierarchical structure of a multi-layer perceptron. The architecture of DR.DEGMON highlights key genes and GO terms that contribute to drug responses through layer-wise relevance propagation (LRP), suggesting potential biological pathways associated with specific drugs. DR.DEGMON achieved a Pearson correlation coefficient of 0.8568 for cell viability prediction, outperforming all baseline models. The model also showed robust generalization performance on external datasets, including GDSC, PRISM, and CCLE. In addition, we employed layer-wise relevance propagation (LRP) to obtain relevance scores for input genes and nodes representing GO terms. DR.DEGMON shows high performance in predicting drug responses and provides interpretable results. The integration of GO and LRP enabled the model to suggest the underlying biological processes involved in drug responses, making it a valuable tool for predicting outcomes and discovering new biomedical knowledge in cancer pharmacogenomics. This approach offers both practical utility in drug development and a method for improving the understanding of cancer biology.

Understanding orchid mycorrhizas and symbiotic germination has long depended on in vitro models, yet these systems fail to capture the environmental forces shaping natural symbiosis. Here, we use RNA-seq to compare symbiotic and asymbiotic protocorms (the post-embryonic organ characteristic of orchid germination) grown in vitro with those developing in the field, allowing us to resolve how ecological context reshapes early plant physiology and plant-fungus interactions. Field-grown protocorms broadly aligned with the in vitro symbiotic model, while also showing how environmental signals refine a shared core transcriptional programme. Both symbiotic conditions showed transcriptomic patterns compatible with fungal colonisation but differed in the relative contribution of regulatory, transport, and stress-associated functions. Field protocorms showed stronger activation of membrane transport and wall-associated mechanosensing pathways, while sustaining defence-related regulation at a restrained, metabolically integrated level. Under in vitro conditions, symbiotic protocorms showed enhanced activation of stress-responsive and detoxification pathways compared with field-grown ones. The expression of light-associated plastid differentiation genes was attenuated despite brief culture-related illumination. In contrast, asymbiotic protocorms showed stronger activation of light- and oxidative-stress-related responses. Together, these patterns indicate that field-grown protocorms show a more integrated and environmentally modulated transcriptional organisation, whereas in vitro symbiosis is associated with stress buffering and compensatory metabolic responses. By integrating field and laboratory datasets, this work provides a more complete portrait of early orchid symbiosis and protocorm physiology, clarifying both the interpretive power and limitations of current in vitro models.

Myocardial injury activates the complement lectin pathway (LP) in acute coronary syndrome (ACS) and LP inhibition improved cardiac function in experimental studies, suggesting a direct pathogenic role. The clinical consequences of LP activation are insufficiently defined. We investigated how plasma levels of the LP activators mannose-binding lectin (MBL) and ficolin-2 (FCN2) relate to cardiac recovery and prognosis in ACS patients. MBL and FCN2 were measured at baseline in a cohort of 546 ACS patients and at 6-weeks in 124 patients with available samples. Prospective associations with heart failure (HF), stroke, and major adverse cardiovascular events (MACE) during a median follow-up of 2.2&#xa0;years were assessed by multivariable Cox regression. Spearman correlation was used to assess relationships between MBL and FCN2 levels, inflammatory and fibrotic mediators in plasma, and echocardiographic parameters of left ventricular (LV) remodelling and dysfunction. Baseline MBL was associated with incident HF (HR 1.50, 95% CI 1.04-2.16, p&#xa0;=&#xa0;0.029), independently of clinical risk factors, revascularization, baseline troponin and renal function. Patients with persistently elevated MBL at baseline and follow-up had increased pro-inflammatory and pro-fibrotic mediators in plasma, dilated LV and reduced LV systolic function at 1-year post-ACS. FCN2 showed no association with the outcomes. ACS patients with persistently high plasma MBL levels suffer cardiac remodelling and dysfunction, and have an increased risk for HF. Our findings provide clinical support to experimental data suggesting that the complement LP might be a potential therapeutic target to prevent post-ACS HF.

The aim of the present study was to describe the association between behavioral and psychological symptoms and psychotropic drug use among older Swedish nursing home residents with cognitive impairment. Data was collected from a questionnaire survey which was sent out between 4 and 12 February 2019 within the Swedish National Inventory of Care and Health in Residential Aged Care (SWENIS) II. Sixty municipalities in Sweden were randomly selected and invited. Of these, 49 municipalities accepted the invitation. Six withdrew resulting in 43 participating municipalities and 187 nursing homes. The final study population comprised 1,541 participants, 65 years or older, with cognitive impairment. The Swedish Prescribed drug register was utilized to identify psychotropic drug use. Backward eliminating binary logistic regression analyses were conducted. At least one psychotropic drug was used of 80.8% of the study population, 20.5% used at least one antipsychotic drug, 32.0% used anxiolytic drugs, 21.3% used hypnotic and sedatives, 58.4% used antidepressants and 32.1% of the residents used antidementia drugs. Among residents with mild cognitive impairment, anxiolytics were significantly associated with anxiety. Also, anxiolytics and hypnotics and sedatives were significantly associated with sleep and nighttime behavior disorders. Antidepressants were significantly associated with depression/dysphoria. The same pattern was found among residents with moderate cognitive impairment. However, anxiolytics were also significantly associated with worsening depression/dysphoria. Hypnotics and sedatives were also associated with anxiety and hallucinations. Moreover, antipsychotic drug use was significantly associated with agitation/aggression and anxiety. Among residents with severe cognitive impairment, antipsychotics, anxiolytics and hypnotics and sedatives were significantly associated with aberrant motor behavior. Anxiolytics were also significantly associated with anxiety and apathy/indifference, and hypnotics and sedatives were significantly associated with agitation/aggression and depression/dysphoria. Psychotropic drug use is remarkably high among Swedish nursing home residents but the prevalence of antidementia drugs is somewhat low. Associations found, call for further studies examining the treatment of behavioral and psychological symptoms (BPS) in this vulnerable group of people.

To predict how movement strategies shape fitness in a changing world, we must build mechanistic models of species' ranges and range shifts that align environmental data with the scales of individual perception and decision-making and jointly model biological processes from the individual to the population scale.

Sabia discolor Dunn is an ethnomedicinally significant plant with underexplored genetic resources. To elucidate its chloroplast genome structure, evolutionary characteristics, and phylogenetic position within the genus Sabia, we sequenced and analyzed its complete chloroplast genome. Using Illumina HiSeq&#x2122; platform for sequencing, followed by assembly with GetOrganelle and annotation with CPGAVAS2, we characterized the genome. The results showed that the chloroplast genome of S. discolor is a 161,581&#xa0;bp circular DNA molecule with a typical quadripartite structure and a GC content of 39.25%. It harbors 129 genes, including 84 protein-coding genes, 8 rRNAs, and 37 tRNAs. We detected 80 simple sequence repeats (SSRs) and 55 long repeats. Codon usage bias analysis revealed a weak preference, with an effective number of codons (ENC) value of 49.01, primarily shaped by natural selection rather than mutation pressure. Comparative analysis identified a significant contraction in the inverted repeat (IR) regions and six highly variable regions (e.g., trnH-psbA, ndhF), which are proposed as potential molecular markers for species identification. Phylogenetic analysis based on complete chloroplast genomes revealed two major clades within Sabia, with S. discolor placed in Clade I, closely clustered with S. parviflora, S. limoniacea, S. fasciculata, and S. swinhoei, while S. dielsii formed a distinct basal lineage. This study provides fundamental genetic resources for future research on the conservation, phylogenetics, and molecular identification of S. discolor and its relatives.

Barley (Hordeum vulgare L.) provides a suitable model for studying domestication-driven plant-microbiome interactions. Although wild, landrace, and modern genotypes host distinct rhizosphere communities, the extent to which roots and microbes reciprocally influence each other remains unclear. Here, we applied an integrated multi-omics approach combining long-read metagenomics, root transcriptomics, and plant genomics to understand genotype-specific host-microbiome coordination. Oxford Nanopore whole metagenome sequencing (WMS) revealed genotype-associated shifts in rhizosphere communities across seasons. Functional profiling showed a conserved metabolic backbone including amino acid metabolism, energy production, and secondary metabolite biosynthesis, alongside genotype-dependent variation in carbohydrate metabolism and transport-associated pathways. Genome-resolved analysis through metagenome-assembled genomes (MAGs) further detailed the taxonomic and functional architecture of key rhizosphere lineages. Root transcriptome profiling identified extensive differential expression associated with microbial perception, signaling, defense, and metabolic processes. Integration of host and microbiome data revealed coordinated molecular patterns, indicating that barley genotypes are associated with distinct microbial assemblages and corresponding transcriptional responses. These findings indicate that domestication has shaped coordinated associations between barley genotypes and their rhizosphere microbiomes, reflected in both microbial community composition and host transcriptional regulation. This work provides new insights into the evolutionary tuning of plant-microbiome relationships and highlights opportunities for microbiome-informed strategies in barley improvement.

Reproductive aging in vertebrates is commonly interpreted as a gradual decline; however, whether aging of the male gonad proceeds linearly or is characterized by periods of accelerated remodeling remains unclear. The turquoise killifish (Nothobranchius furzeri), a short-lived vertebrate, enables lifespan-wide resolution of both rapid germline establishment and subsequent aging within months. Our staged histological atlas shows that the testis progresses from a simple gonadal primordium to completed spermatogenesis within 3 to 4&#xa0;weeks post-hatching, revealing an exceptionally compressed developmental program. Proliferative activity peaks in early adulthood, marking maximal spermatogenic output, but declines sharply at mid-age. This period coincides with coordinated repression of germline, mitotic, and meiotic programs and activation of extracellular matrix remodeling, angiogenic, inflammatory, and stromal pathways. These molecular shifts are accompanied by structural remodeling, including expansion of the interstitial compartment and accumulation of collagen-rich matrix. Late-life testes exhibit comparatively modest additional changes, suggesting stabilization of a remodeled, low-proliferative niche. Testicular aging in the turquoise killifish unfolds as a process marked by a pronounced mid-age remodeling shift that links testicular decline to somatic niche remodeling. This remodeling window provides a mechanistic entry point for dissecting reproductive aging in a short-lived vertebrate, with potential relevance for comparative studies in other vertebrate systems, although these dynamics may be shaped by species-specific life histories. Together, these findings establish the killifish as a uniquely powerful model for identifying interventions that preserve germline function.

Salicylic acid (SA) and benzoic acid (BA) are important phenolic regulators involved in plant growth and defense; however, their comparative roles in modulating growth and secondary metabolites production in leafy vegetables remain unclear. Therefore, this study aimed to investigate the influence of exogenous SA and BA on growth, mineral composition, and bioactive compound accumulation in Swiss chard (Beta vulgaris L. var. cicla). Field experiments were conducted over two successive seasons of 2023/24 and 2024/25 to assess the influence of SA and BA on Swiss chard cv. Ruby Red. Plants were treated with SA and BA at 1, 2, and 3 mM, each alongside an untreated control. Treatments were arranged in a randomized complete block design with three replicates. Beside the agronomic attributes, photosynthetic pigments, total phenols, total flavonoids, total antioxidant activity, total indoles, anthocyanin, nitrate, and mineral content were assayed. Both SA and BA significantly improved growth parameters, mineral content, and secondary metabolites accumulation compared to the control. SA at 1 mM or BA 1 mM were the most effective treatments, showing the highest significant values of vegetative parameters, photosynthetic pigments, total phenolics, total flavonoids, total antioxidant activity, total indoles, anthocyanin, and mineral content. Further, BA 1 mM was the potent treatment for lowering leaf nitrate content, showing the lowest nitrate content, which was significantly lower than SA at 2 and 3 mM. Correlation analysis revealed strong positive relationships between growth parameters and secondary metabolites. BA behaves influence similar to SA for enhancing leaves yield with higher efficacy in reducing nitrate content via enhancing the efficiency of photosynthetic pigments and regulating various secondary metabolites. Herein, BA at low concentration (1 mM) is a promising rate to be applied to obtain high yield with good quality.

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Single-cell foundation models such as scGPT and Geneformer learn rich representations of gene expression programs, but whether these representations encode gene regulatory relationships beyond expression-level confounds remains unclear. Attention patterns in these models have been shown to capture co-expression rather than direct regulation, leaving open the question of whether deeper representations-particularly the residual stream-contain genuine regulatory information. We systematically investigated residual-stream geometry in scGPT and Geneformer across four tissue contexts from the Tabula Sapiens atlas, evaluating whether geometric proximity between gene vectors provides incremental predictive value for curated TRRUST transcription factor-target edges beyond expression confounds. Under repeated stratified cross-validation, geometric features provided significant incremental signal in kidney and immune settings, validated by label-permutation and geometry-shuffle null controls; centered-cosine similarity, PCA projection and multi-layer bundling recovered comparable signal in lung tissues, and the multi-layer bundle improved every domain (kidney &#x394;AUROC&#xa0;=&#xa0;+&#xa0;0.122, immune +&#xa0;0.042, lung +&#xa0;0.028, external lung +&#xa0;0.027; geometry-augmented AUROC 0.60-0.69). The effect was fully robust to leave-TF-out and leave-target-out cross-validation and to harder degree- and expression-matched negative edges, but under the stricter leave-both-out split-no transcription factor and no target shared between folds-it collapsed to near-zero (&#x394;AUROC at most +&#xa0;0.003, and not statistically significant in kidney or immune), marking the ceiling of out-of-entity generalization. With a comparable per-layer residual-stream extraction applied to both models, the apparent Geneformer advantage mostly disappeared (small residual gaps remained in three of four domains), indicating it largely reflected representation-construction choices rather than a substantial architectural difference. Asymmetric geometric features predicted regulatory edge orientation (AUROC 0.80-0.90), and the geometric signal added incremental value on top of expression-based gene regulatory network (GRN) inference (GENIE3, co-expression). Foundation model residual streams carry incremental, regulatory-relevant geometric signal that is distributed across layers and that complements expression-based GRN inference for retrospective edge prioritization. The signal is statistical enrichment rather than a stand-alone regulatory classifier: absolute performance is modest and out-of-entity generalization is limited, so its practical role is as an orthogonal evidence channel for edge re-ranking and hypothesis prioritization in multi-evidence frameworks.

Bisphosphonates (BPs) are widely used antiresorptive agents for skeletal disorders; however, their adverse effects on oral soft tissues, particularly the lingual mucosa and gustatory apparatus, remain incompletely understood. Nano-chitosan has emerged as a promising biomaterial with cytoprotective and regenerative properties in orofacial tissues, indicating potential utility in mitigating BP-associated mucosal injury. To evaluate the protective effects of topical nano-chitosan on alendronate-induced lingual injury in a rat model. Thirty adult male Wistar rats were randomized into three groups (n&#x2009;=&#x2009;10 each): control (Group I), alendronate-treated (1&#xa0;mg/kg, subcutaneously, three times weekly for 4 weeks), and alendronate plus daily topical nano-chitosan (0.5 mL/day). Lingual tissues were examined using histology (H&E and Masson's trichrome), immunohistochemistry (PCNA), histomorphometry, scanning electron microscopy, and RT-qPCR for &#x3b1;-gustducin (GNAT3). Alendronate induced significant lingual alterations, including reduced papillary height (48%), decreased epithelial proliferation (80% PCNA expression), increased disorganized collagen deposition (approximately 2.5-fold), ultrastructural disruption, and downregulation of GNAT3 (all p&#x2009;<&#x2009;0.001 vs. control). Nano-chitosan treatment attenuated these changes, restoring papillary morphometry, PCNA expression (27.66% vs. 29.60%, p&#x2009;=&#x2009;0.127), and collagen content (9.45% vs. 8.49%, p&#x2009;=&#x2009;0.538) to levels comparable with controls. GNAT3 expression was partially restored but remained significantly reduced compared with controls. Topical nano-chitosan mitigates alendronate-induced lingual injury in rats by improving epithelial architecture and proliferation and partially restoring GNAT3 transcript levels. No functional gustatory assessment was performed; therefore, conclusions regarding taste function recovery are not drawn from the present data. These findings indicate a modulatory rather than fully restorative effect on BP-associated oral mucosal alterations and support further preclinical and translational investigation.

The "nose-brain axis" has been proposed as a key mechanism linking allergic rhinitis (AR) to central nervous system (CNS) dysfunction; however, alterations in functional connectivity (FC) gradients remain unexplored in AR. This cross-sectional study aimed to investigate the correlation between brain FC gradients and AR-related peripheral multi-omics profiles, as well as their clinical significance, through multimodal cross scale analysis. We enrolled cross-scale data from 22 AR patients and 20 healthy controls (HCs), including resting-state functional magnetic resonance imaging (rs-fMRI), serum proteomics and metabolomics, and clinical assessments. FC gradient analysis was employed to investigate hierarchical brain functional organization, with gradient values compared at global, regional, and network levels. Multivariate partial least squares (PLS) analysis was used to integrate the multidimensional associations among FC gradients, peripheral molecular signatures, and behavioral phenotypes. No significant differences were observed at the global or Yeo's seven canonical networks. However, refined regional analysis revealed significant FC gradient alterations predominantly within the Default mode (DMN), Somatomotor (SMN), and Limbic (LN) networks, particularly in RH_Default_Temp_5 (p&#x2009;=&#x2009;0.0031) and LH_Limbic_OFC_4 (p&#x2009;=&#x2009;0.0034). These regional gradient abnormalities correlated significantly with AR severity and neuropsychiatric symptoms (p&#x2009;<&#x2009;0.05), indicating "local fine-tuning" rather than "global collapse" in brain functional remodeling. Multi-omic profiling identified 121 differentially expressed proteins, 197 positive-ion-mode metabolites, 134 negative-ion-mode metabolites. The integrative analysis demonstrated significant associations between these peripheral molecular signatures and AR-related FC gradient alterations. These differentially expressed molecules were primarily enriched in immune response, complement activation, lipids and lipid-like molecules, and cholesterol metabolism pathways. This study provides the first evidence of distinct FC gradient alterations in AR that are coupled to peripheral multi-omic shifts, offering novel insights into CNS mechanisms of AR and identifying potential molecular-neuroimaging biomarkers candidates for precision diagnosis and targeted therapy.

Dental students must master endodontic anatomy, radiographic diagnostics, and treatment strategies early in their clinical training-yet traditional teaching methods, including two-dimensional radiographs, often fail to effectively convey complex spatial structures. Hologram technology has emerged as a promising tool for three-dimensional visualization in dental education. The aim of the study was to compare the classification accuracy of dental students using holograms versus conventional periapical radiographs in identifying endodontic structures. This randomized single center-controlled trial included 79 dental students (years 4-6). After a calibration seminar and pre-test each of the 79 students assessed six tooth cases-three presented as holograms and three as periapical radiographs. Holograms were created from cone-beam computed tomography data, annotated by two calibrated examiners to establish reference points regarding the number of canals, Vertucci classification, and treatment complexity. Students evaluated each case for these parameters; responses were compared to the reference points to assess classification accuracy. The final evaluation form captured their subjective perceptions of the new hologram system. Primary binary outcomes were analyzed using logistic mixed-effects models. P-values were adjusted for multiple testing using the Benjamini-Hochberg procedure. Statistical significance was set at p&#x2009;<&#x2009;0.05. After Benjamini-Hochberg correction, holography significantly improved root-canal number identification in three multi-rooted cases (adjusted p&#x2009;&#x2264;&#x2009;0.007). For Vertucci classification, only the maxillary anterior tooth remained significant (adjusted p&#x2009;=&#x2009;0.044). Treatment-complexity assessment showed case-dependent results, favoring radiography in two cases (adjusted p&#x2009;=&#x2009;0.045 and p&#x2009;=&#x2009;0.006) and holography in one case (adjusted p&#x2009;=&#x2009;0.013). Holographic visualization demonstrated case- and task-specific effects in endodontic training. Holograms were able to improve classification accuracy in certain endodontic tasks. However, they did not outperform conventional periapical radiographs in all outcomes. Holography could be an effective, supplementary tool for teaching certain endodontic tasks, but its application requires further investigation. The study was prospectively registered in the German Clinical Trials Register (DRKS-ID: DRKS00035000) on 13.09.2024.

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