Background: The rising consumption of Haliotis discus hannai (HDH) has brought its associated food allergy into focus. However, studies on its allergens and their role in patient reactions remain limited. This study aimed to construct a high-abundance allergen profile for HDH by integrating multi-omics data and to identify its immunodominant allergens through immunobinding assays with volunteers’ sera. Methods: We first constructed a predicted proteome for HDH using in-house generated transcriptome data along with public genome resources. High-throughput proteome and quantitative expression data were employed to establish a high-abundance allergen profile. Immunobinding assays, incorporating Western-blot analyses with volunteer-derived sera, were utilized to delineate allergens characterized by high immunoreactivity. Their tertiary structures and B-cell epitopes were subsequently analyzed using multiple bioinformatics tools. Results: We predicted 53,245 protein-coding genes from a high-quality assembled genome and transcriptome, of which 33,109 were functionally annotated. Proteome sequencing identified 3,916 reliably expressed proteins. Cross-referencing with two authoritative allergen databases revealed 37 high-abundance allergens in HDH. Sequence comparisons indicated a high similarity for homologous allergens between HDH and other aquatic mollusks, crustaceans, and terrestrial arthropods, indicating a potential widespread cross-reactivity. Dot-blot and Western-blot assays using sera from 90 volunteers identified six protein bands with significant IgE-binding activity. Based on molecular weight and prior allergen profiling, the six bands were hypothesized to be a polymer, paramyosin (PM), arginine kinase (AK), tropomyosin (TM), triosephosphate isomerase (TIM), and fructose-bisphosphate aldolase (FBA), respectively. Subsequent immunobinding assays identified PM, AK, TM, TIM, and FBA as the immunodominant allergens, as they each exhibited relatively high seropositivity rate in volunteers’ sera. Treatment with denaturants (SDS and β-me) revealed that PM and TM are primarily linear allergens, whereas AK is predominantly conformational, explaining the higher detection rate in Dot-blot versus Western-blot. Homology modeling showed that PM and TM possess relatively simple structures dominated by α-helices, while AK, TIM, and FBA have more complex tertiary structures containing significant proportions of β-sheets and random coil. B-cell epitope prediction indicated that PM and TM harbor mainly linear epitopes, whereas AK, TIM, and FBA possess additional conformational epitopes. Conclusions: This study provided the first comprehensive allergen profile for HDH by integrating multi-omics data and immunoassays, and further identified its immunodominant allergens. Through bioinformatics predictions and denaturant experiments, we elucidated the structural conformations and B-cell epitopes of immunodominant allergens. These findings offer crucial data for the precise identification of abalone allergens, supporting the development of accurate diagnostic strategies for affected individuals.

Melanoma in Asia presents a unique epidemiological profile, with a higher prevalence of acral and mucosal subtypes compared to Western populations. While KIT mutations are found in up to 15% of Asian melanoma cases, clinical outcomes with KIT inhibitors have been modest due to heterogeneous mutation profiles and a lack of specific patient selection criteria. This study characterizes the landscape of KIT mutations in melanoma using the GENIE database, identifying 86 recurrent hotspots, many of which are variants of unknown significance (VUS). We validated drug sensitivities for key mutations using in vitro and in vivo models. Our results indicate that while the L576P mutation is highly sensitive to multiple inhibitors, the N822K mutation shows resistance to imatinib but responds to sunitinib, nilotinib, and nintedanib. These findings highlight the necessity of genotype-guided therapeutic strategies and provide a rationale for future clinical trials combining broad-spectrum KIT inhibitors with immune checkpoint inhibitors. Melanoma subtypes prevalent in Asia, specifically acral and mucosal melanoma, frequently harbor KIT mutations but show poor response rates (23-26%) to the standard-of-care inhibitor, imatinib. This study challenges the current clinical practice of treating all KIT -mutated melanomas uniformly. We demonstrate that specific recurrent mutations, such as N822K, are intrinsically resistant to imatinib but highly sensitive to broad-spectrum inhibitors like sunitinib and nintedanib. By establishing a comprehensive “lookup table” of drug sensitivities for both common and previously uncharacterized KIT variants, this work provides the evidence base required to transition from a “one-size-fits-all” approach to a genotype-guided precision medicine strategy. Furthermore, validating these targets informs the design of next-generation clinical trials, particularly those combining optimal KIT inhibitors with immune checkpoint blockade to improve survival in currently underserved patient populations.

The transition from anaerobic to aerobic life was a pivotal adaptation in Earth's history, yet the timing and genomic drivers remain poorly resolved. Traditional approaches relying on oxygen-utilizing genes need improvement for obligate anaerobes and fragmentary environmental genomes, where gene absence may reflect poor assembly rather than phenotype. We developed a machine learning model (GBDT40-LR) to predict microbial oxygen requirements using 40 broadly conserved genes, 35 without direct oxygen roles. This approach overcomes incompleteness biases in environmental genomes. Applied to 80,787 bacterial genomes [including metagenome-derived assemblies (MAGs)], the model classified 42,014 aerobes and 38,775 anaerobes, enabling large-scale ancestral reconstruction. Molecular clock dating indicates an emergence of aerobic bacterium prior to the Great Oxidation Event (GOE, 2.5 to 2.3 Ga), likely around ~2.7 Ga. Aerobic lineages subsequently diversified during the GOE and Neoproterozoic Oxygenation Event (NOE, 0.8 to 0.55 Ga), with persistent anaerobe diversity across Earth's oxygenation. This establishes that aerobic bacteria originated planetary oxygenation, potentially by 200 to 400 My, providing insights into phenotypic evolution and prolonged anaerobe-aerobe coexistence.

Amidst the exponential rise in crustacean production and consumption across East Asia, the population burden of crustacean allergies has intensified correspondingly. Therapeutic options remain limited, and contemporary risk stratification still relies predominantly on familial history, an approach whose predictive granularity is inadequate for precision public health. Converging evidence from twin and family cohorts indicates that crustacean allergies exhibit a high narrow-sense genetic heritability, underscoring the imperative to dissect its genomic architecture and to quantify downstream health sequelae. Here, we conducted a comprehensive review with an integrated methodological perspective, mainly synthesizing recent advances in the molecular genetics of crustacean allergies and delineating their translational ramifications for early-life risk prediction. This review primarily focused on the high prevalence of crustacean allergies in East Asia and the inadequacies of current early risk assessment systems, genetic loci associated with crustacean allergies and existing gaps in molecular genetic studies, the applications of polygenic risk scores in early risk assessment, and the evaluation of genetic causal effects of crustacean allergies on the risk of human common diseases. Leveraging two-sample Mendelian randomization anchored in large-scale East Asian genome-wide association study summary statistics, we demonstrated that genetically predicted shrimp allergy exerted significant causal effects on a spectrum of human common disorders spanning the respiratory, gastrointestinal, endocrine, musculoskeletal, nervous, cardiovascular, integumentary, and autoimmune systems. These findings underscored the necessity for comprehensive molecular genetic studies to develop an objective assessment framework that facilitates early identification of high-risk populations and evaluate their adverse effects on individual’s health, prioritizing precision immunomodulatory regimens targeting high-risk individuals before clinical manifestation.

Purpose: To investigate whether secondhand smoke (SHS) exposure alters the ocular surface microbiome (OSM) in children and to explore potential functional consequences. Methods: 432 children aged 3-18 years were enrolled, including 111 SHS-exposed and 321 unexposed controls. Conjunctival swabs were collected and analyzed by 16S rRNA gene sequencing targeting the V3-V4 region. Sequencing data were processed with Qiime2 and DADA2, and taxonomic classification was based on the SILVA 138 database. Alpha diversity and beta diversity were compared using t-tests and PERMANOVA. Differentially abundant taxa were identified using LEfSe, and predicted functional pathways were analyzed using PICRUSt2 with MetaCyc and KEGG annotation. Results: SHS-exposed children showed significantly altered alpha diversity (Chao1, Shannon, Simpson) and distinct beta diversity compared with controls. LEfSe analysis revealed enrichment of several phyla and genera, including Lactobacillus and Rubellimicrobium in controls, with no taxa enriched in SHS-exposed children. Functional prediction showed enrichment of metabolism pathways such as L-methionine salvage, biphenyl, heparin, and toluene degradation and immune-related pathways, including complement activation, T and B cell receptor signaling, MAPK, and TGF-beta pathways. Conclusion: SHS exposure in children is associated with significant alterations in ocular surface microbial diversity, community structure, and predicted functional pathways related to environmental stress and immune signaling. These findings highlight the sensitivity of the pediatric OSM to SHS exposure and underscore the importance of minimizing environmental tobacco smoke to protect children's ocular health.

Epigenetic inheritance is fundamental to human development and disease, yet the mechanisms governing the transmission of DNA methylation across generations remain incompletely understood. In this study, we performed haplotype-resolved, whole-genome DNA methylation profiling in a healthy three-generation Chinese family, leveraging high-depth Oxford Nanopore Technologies (ONT) and PacBio HiFi long-read sequencing, anchored to a proband-specific telomere-to-telomere (T2T) genome assembly. We observed globally conserved bimodal methylation landscapes across all individuals and generations. Stratified analyses revealed clear functional compartmentalization of methylation marks, characterized by distinct hypomethylation in centromeres and hypermethylation in retrotransposons and repetitive elements. Chromosome-resolved analysis of ribosomal DNA (rDNA) arrays demonstrated a domain-specific methylation pattern with hypomethylation in the transcriptional core and hypermethylation in the intergenic spacer, with evidence for age-associated epigenetic drift in the transcriptional core domain. Through de novo identification and validation, we mapped 23 high-confidence imprinting control regions (ICRs) showing robust parent-of-origin-specific methylation, all overlapping known imprinted genes and enriched for regulatory element signatures. Haplotype-resolved X chromosome analysis further uncovered sex- and allele-specific methylation patterns linked to X inactivation dynamics. Together, this pedigree-scale, high-resolution study delineates the landscape and principles of intergenerational DNA methylation inheritance, revealing both conserved and dynamic features shaping the human epigenome.

This study aims to differentiate between unipolar and bipolar depressive episodes through an integrated analysis of gut microbiome and serum metabolome. The study involved 82 patients experiencing depressive episodes, with 38 diagnosed with Major Depressive Disorder (MDD) and 44 with Bipolar Disorder (BD). The gut microbiome and serum metabolome were analyzed using 16S rRNA sequencing and ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MS), respectively. The results revealed distinct microbial compositions and metabolic pathways between the two groups. Seventeen microbial groups and fifty serum metabolites were found to be significantly different between the two groups. Four genera and eight serum metabolites demonstrated strong diagnostic potential for differentiating BD from MDD. The study also found correlations between certain differential genera and metabolites and the severity of clinical symptoms. This integrated multi-omics approach provides a promising direction for the differential diagnosis of unipolar and bipolar depression.

Endothelial and macrophage inflammation and sterol transport play an important role in atherogenesis. Serum/glucocorticoid-regulated kinase 1 (SGK1) is a member of the serine/threonine kinase family sharing approximately 54% identity with Akt. It has been implicated in smooth muscle cell calcification and macrophage inflammation during atherosclerosis; however, the role of SGK1 in endothelial dysfunction and endothelial or macrophage lipid metabolism is less characterized. In this study, we intraperitoneally injected high-cholesterol diet-fed male ApoE-knockout mice with the SGK1 inhibitor EMD638683 (10 mg/kg) every other day for 2 weeks, followed by histopathological and transcriptome analysis of the atherosclerotic lesion. To study the SGK1-associated mechanism in vascular inflammation in vitro, SGK1 silencing was performed in primary human umbilical vein endothelial cells (HUVECs), followed by treatment with pro-inflammatory cytokines. In THP-1-differentiated macrophages, SGK1 silencing followed by treatment with lipopolysaccharides was used. We showed that ApoE-knockout mice treated with EMD638683 show reduced atherosclerotic plaque area and attenuated endothelial and macrophage inflammation. Further transcriptome analysis of thoracic aortae showed that SGK1 inhibition downregulated inflammation and lipid metabolism-associated genes. In vitro, interleukin-1β treatment induces SGK1 phosphorylation. SGK1 inhibitor or siRNA reduced endothelial inflammation induced by pro-inflammatory cytokine treatments in HUVECs through nuclear factor κ light chain enhancer of activated B cell (NF-κB) signaling. In THP-1-differentiated macrophages, SGK1 inhibition or knockdown by siRNA is associated with reduced levels of pro-inflammatory cytokines, NF-κB, and sterol regulatory element binding protein 1 (SREBP1) pathway activation, following lipopolysaccharide treatment. SREBP1 inhibition by fatostatin or siRNA reduced p65 phosphorylation. In conclusion, the inhibition of SGK1 has been shown to reduce atherosclerotic plaque area and attenuate endothelial and macrophage inflammation in ApoE-knockout mice via NF-κB and SREBP1 signaling. These results not only contribute to our understanding of the complex interplay between vascular inflammation and lipid metabolism but also hold promise for the development of novel therapeutic strategies for atherosclerosis.

Large language models can learn new tasks through in-context learning (ICL), yet this ability remains underexplored for biological sequence classification. We evaluate ICL across 20 large language models on three antibody tasks: species-origin, antibody specificity, and isotype class classification. Few-shot prompting improves over zero-shot performance, but matching the performance of protein language model classifiers requires sequence-similar demonstrations. Building on this observation, we introduce a sequence similarity-based strategy for ICL in antibody sequence classification, Sim-ICL. Using 32-shot prompting, Sim-ICL achieves competitive performance on two of three tasks. Its simplicity makes few-shot ICL promising for antibody characterization, especially for researchers with limited coding expertise.

NRAS mutations drive 20–30% of melanomas and are associated with poor response to immunotherapies. RAS(ON) inhibitors such as daraxonrasib (RMC-6236) have shown promising activity in NRAS-mutant melanoma, yet mutation-specific sensitivity, resistance mechanisms, and rational combination strategies remain largely unresolved. Here, we performed saturation mutagenesis across NRAS mutation hotspots to systematically evaluate mutation-resolved oncogenic fitness and drug responsiveness to six direct RAS inhibitors (sotorasib, adagrasib, ADT-007, BI-2865, RMC-6236, and RMC-7977) using isogenic melanoma spheroids, xenografts, and pooled drug-sensitivity profiling. RMC-6236 and RMC-7977 displayed the broadest activity, sensitizing ∼80% of recurrent NRAS mutations across the P-loop and Switch II regions. Several clinically observed mutations, including Q61P, G13D/V/R, and G60E, showed reduced susceptibility to RAS(ON) inhibition. Structural modelling and thermostability analysis indicated impaired inhibitor engagement or reduced tricomplex stability as potential mechanisms of resistance. RAS(ON) blockade also induced STAT3 activation, particularly in oncogenic Q61 mutants. Co-targeting STAT3 with napabucasin or siRNA enhanced apoptosis, suppressed MYC, and improved tumour control in vitro and in vivo. Together, this study provides the first comprehensive mutation-resolved sensitivity atlas for NRAS-mutant melanoma, highlights structural mechanisms underlying resistance to RAS(ON) inhibitors, and identifies STAT3 inhibition as a rational combination strategy. These findings offer preclinical evidence supporting mutation-guided patient stratification and combination-therapy development for NRAS-mutated melanoma. NRAS-mutated melanoma lacks effective targeted treatments, and clinical responses to immunotherapy remain suboptimal. This study presents the first comprehensive drug sensitivity map across 95 NRAS mutations in melanoma, identifying the pan-RAS(ON) inhibitors RMC-6236 and RMC-7977 as broadly effective agents. Multiple mutants with reduced susceptibility are identified, providing mutation-specific guidance for patient selection and clinical trial stratification. Mechanistic analyses reveal that STAT3 activation functions as a key survival pathway under RAS(ON) blockade, and its inhibition markedly enhances the efficacy of pan-RAS(ON) inhibitors. These findings support mutation-guided utilization of RAS(ON) inhibition and highlight STAT3 co-targeting as a rational strategy to strengthen and prolong therapeutic responses in NRAS-mutant melanoma. - Generation of mutation-resolved functional and drug-sensitivity atlas for 95 NRAS missense variants covering >99% of all NRAS recurrent mutation - RMC-6236 and RMC-7977 shows broad and potent activity across major NRAS hotspot mutations - Defines NRAS mutations with reduced susceptibility to provide actionable information for clinical trial stratification - RAS(ON) inhibition and napabucasin shows synergistic anti-tumor activity in NRAS-mutated melanoma

Abstract Immune checkpoint inhibitor (ICI) resistance in non-small cell lung cancer (NSCLC) is frequently driven by an immunosuppressive tumor microenvironment (TME). To address this issue, we developed IL-24-iMSC, a novel mesenchymal stem cell-based therapy designed to remodel the immunosuppressive tumor microenvironment through sustained secretion of interleukin 24 (IL-24), an immunomodulatory cytokine with anti-cancer properties. Induced mesenchymal stem cells (iMSCs) were derived from induced pluripotent stem cells (iPSCs) and the IL-24 gene was integrated into iMSC at the B2M locus under the control of the EF1α promoter using CRISPR/Cas. IL-24-iMSCs were confirmed to constitutively secrete IL-24. Metastatic lung cancer models were established by tail vein injection of Lewis Lung Carcinoma cells expressing luciferase (LLC-luc) into immunocompetent C57BL/6 mice. Tumor progression was monitored using an In Vivo Imaging System (IVIS). Treatment was administered 20 days after tumor inoculation, and mice were randomized into three groups: iMSC, IL-24-iMSC, or vehicle control. All mice were sacrificed three days after treatment. Tumor growth was confirmed by bioluminescence imaging using IVIS, and tumor samples were collected for immune profiling. While there was no significant difference in tumor growth among the three groups, tumors exposed to IL-24-iMSC demonstrated a pro-inflammatory phenotype. Multiplex immunohistochemistry confirmed infiltration of MSCs into the TME, with significantly increased IL-24 expression in the IL-24-iMSC arm. Compared with the control group (untreated), tumors treated with IL-24-iMSC showed increased CD8+ T cells, dendritic cells, and memory CD4+ T cells. IFN-γ and TNF-α expressions were also elevated in the IL-24-iMSC treatment arm. On the contrary, regulatory T cells and exhausted T cells (PD1+ TIM3+) were significantly reduced in the IL-24-iMSC group. In conclusion, IL-24-iMSCs demonstrated the ability to modulate immune cell composition within the tumor microenvironment, suggesting their potential to synergize with ICI in NSCLC. Citation Format: Yuting Zhang, Yuxuan Zhang, Sai Fung Yeung, Chi-Hang Wong, Tsz Tung Kwong, Connie Wun Chun Hui, Stephen Kwok-Wing Tsui, Tony S. Mok, Desheng Liang, Molly Siu Ching Li. IL-24 engineered mesenchymal stem cells as a novel therapeutic strategy to remodel the tumor microenvironment in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 7934.

Summary Mutations in Cyclin-Dependent Kinase-Like 5 (CDKL5) cause CDKL5 deficiency disorder (CDD), an X-linked neurodevelopmental condition. Through a phosphoproteomic screen, we identified the neuron-specific nELAVL family of RNA-binding proteins as direct activity-dependent substrates of CDKL5. In support of this regulatory axis, single-nuclei transcriptomics of Cdkl5 knockout (KO) cortices revealed an enriched reduction in activity-dependent mRNAs. Mechanistically, we show that nELAVL proteins undergo phase separation to form biomolecular condensates, the size of which is gated by CDKL5 phosphorylation. Loss of CDKL5 leads to enlarged nELAVL condensates, which exhibit reduced binding affinity for the target mRNA Fos , resulting in its accelerated degradation. This disruption extends to inter-condensate communication: phosphodeficient nELAVL show diminished interaction with P-bodies, which themselves become enlarged in CDD mutant iNeurons. Functionally, the absence of nELAVL phosphorylation recapitulates the deficits in experience-dependent visual function observed in Cdkl5 KO mice. Our findings establish a critical molecular mechanism by which CDKL5-mediated phosphorylation governs mRNA metabolism by tuning the properties of nELAVL condensates and their communication with other biomolecular condensates, ultimately promoting experience-dependent maturation of the visual cortex.

Abstract Primary human hepatocytes (PHHs) are the gold standard for toxicology and drug metabolism studies in industry. However, their limited availability, substantial batch-to-batch variability, and high cost restrict their use. Here, we report a novel culture condition that reprograms PHHs into a proliferative state. These proliferating cells, termed precursors of chemically expanded hepatocytes (pre-cHep), expand over 10 6 -fold within 30 days while retaining liver repopulation capacity comparable to PHHs. pre-cHep can further differentiate into chemically expanded hepatocytes (cHep) as three-dimensional (3D) spheroids within 7 days in vitro , exhibiting global gene expression profiles, albumin production, and cytochrome P450 (CYP) activities similar to 3D-cultured PHH spheroids (3D PHH). Efficient genetic manipulation of pre-cHep using CRISPR/Cas9 is also achievable. Together, pre-cHep and cHep represent a promising alternative to high-quality PHHs, providing a more affordable, reproducible, and scalable source of human hepatocytes for toxicology, drug metabolism studies, disease modelling, towards precision drug development.

Background Asian carps are popular freshwater fish species in Hong Kong and served as an important protein source for infants. Grass carp parvalbumin was reported to have higher allergenicity than other commercially important fishes (salmon and cod) in terms of IgE reactivity. Objective To identify allergens in Asian carps and study the divergence pattern of allergens. Method Three complete Asian carps’ genomes were constructed by hybrid assembly approach. All the homologs of known and putative allergens were identified based on sequence homology at the genomic level. The gene structures, expression level and protein conformations of parvalbumin were revealed using bioinformatics tools. Results Parvalbumin is the major allergen in grass carp tested by indirect ELISA. Nine homologs of parvalbumin were identified in the genome of grass carp, in which Cid_PV2 exhibited highest sequence similarity to the reported group 1 allergen of grass carp, Cten i 1. Similarly, ten parvalbumin homologs were identified in the genome of bighead and black carp respectively, in which Mpi_PV7 and Hno_PV7 were identified as the putative allergenic homologs. The expression profile of parvalbumin revealed alternative usage of homologs in different tissues of Asian carps. Moreover, gene copy numbers of parvalbumin gradually expanded in bony fish and two gene clusters of parvalbumin were identified inside the genomes of Asian carps. Conclusion All the homologs of Asian carps’ parvalbumin were accurately identified and gene divergence contributed to the formation of allergenic homologs. Together with a comprehensive gene sequence profile of carps’ parvalbumin, those could be applied to achieve a more precise clinical diagnostic test and would ultimately assist the design of preventive treatment by immunotherapy of fish allergy.

PURPOSE:Crustacean shellfish is one of the eight most common food allergens, and crayfish is a highly valued shellfish species for consumption in China. However, the detailed allergen profile of crayfish remains unknown, with only four allergen groups reported in the WHO/IUIS allergen nomenclature database. In this study we aimed to identify novel allergens based on the Procambarus clarkii genome and to reveal its allergen profile for developing better diagnostic tools and treatments. METHODS:We assembled the crayfish genome using both long-read and short-read sequencing data and identified putative allergens using the BLAST algorithm based on sequence homology. We employed bioinformatics tools to investigate the expression levels, gene structure, and synteny of these putative allergens. We also applied indirect enzyme-linked immunosorbent assay by using patients' sera to determine allergenicity and utilized proteomic methods to identify novel allergens. RESULTS:We identified a total of 11 putative allergen groups, including all isoforms or homologs for each allergen group based on the genome and three putative allergens by using 2-dimensional (2D) mass spectrometry. We identified 2 novel allergens, pPro c 3.0301 and pPro c 6.0201, with immunoglobulin E reactivity of 33.3% and 20%, respectively. CONCLUSIONS:By providing a comprehensive understanding of the complete allergen profile, our study presents a foundation for comprehending P. clarkii-associated allergy. The knowledge could facilitate the implementation of a components-resolved diagnostic test and preventive immunotherapy based on molecular allergens for crayfish allergy.

Haliotis discus hannai tropomyosin (HTM), Alectryonella plicatula tropomyosin (ATM), and Mimachlamys nobilis tropomyosin (MTM) were reported as significant seafood allergens capable of eliciting severe allergic reactions. However, studies of cross-reactivity and epitope mapping among them are notably limited. This study discovered their cross-reactivity property through inhibition of IgG/IgE binding capacity and promotion of specific CD4+T-cell proliferation. For T-cell epitopes, 3, 4, and 3 epitopes were identified from the splenocytes of mice immunized with HTM, ATM, and MTM, respectively. In terms of B-cell epitopes, a combination of bioinformatics techniques and serological assays identified 4, 3, and 4 epitopes for HTM, ATM, and MTM, respectively. Sequence logo analysis revealed 2 conserved T-cell epitopes and 5 conserved B-cell epitopes, which may be critical for the observed cross-reactivity property. In conclusion, these findings provide crucial molecular evidence that could enhance the prevention of allergic reactions to aquatic mollusks.

BACKGROUND:The Hong Kong orchid tree Bauhinia × blakeana Dunn has long been proposed to be a sterile interspecific hybrid exhibiting flower heterosis when compared to its likely parental species, Bauhinia purpurea L. and Bauhinia variegata L. Here, we report comparative genomic and transcriptomic analyses of the 3 Bauhinia species. FINDINGS:We generated chromosome-level assemblies for the parental species and applied a trio-binning approach to construct a haplotype-resolved telomere-to-telomere (T2T) genome for B. blakeana. Comparative chloroplast genome analysis confirmed B. purpurea as the maternal parent. Transcriptome profiling of flower tissues highlighted a closer resemblance of B. blakeana to its maternal parent. Differential gene expression analyses revealed distinct expression patterns among the 3 species, particularly in biosynthetic and metabolic processes. To investigate the genetic basis of flower heterosis observed in B. blakeana, we focused on gene expression patterns within pigment biosynthesis-related pathways. High-parent dominance and overdominance expression patterns were observed, particularly in genes associated with carotenoid biosynthesis. Additionally, allele-specific expression analysis revealed a balanced contribution of maternal and paternal alleles in shaping the gene expression patterns in B. blakeana. CONCLUSIONS:Our study offers valuable insights into the genome architecture of hybrid B. blakeana, establishing a comprehensive genomic and transcriptomic resource for future functional genetics research within the Bauhinia genus. It also serves as a model for exploring the characteristics of hybrid species using T2T haplotype-resolved genomes, providing a novel approach to understanding genetic interactions and evolutionary mechanisms in complex genomes with high heterozygosity.