Genetically engineered CD276-anchoring biomimetic nanovesicles target senescent escaped tumor cells to overcome chemoresistant and immunosuppressive breast cancer.

Chemotherapy-induced cellular senescence leads to an increased proportion of cancer stem cells (CSCs) in breast cancer (BC), contributing to recurrence and metastasis, while effective means to clear them are currently lacking. Herein, we aim to develop new approaches for selectively killing senescent-escape CSCs. High CD276 (95.60%) expression in multidrug-resistant BC cells, facilitates immune evasion by low-immunogenic senescent escape CSCs. CALD1, upregulated in ADR-resistant BC, promoting senescent-escape of CSCs with an anti-apoptosis state and upregulating CD276, PD-L1 to promote chemoresistance and immune escape. We have developed a controlled-released thermosensitive hydrogel containing pH- responsive anti-CD276 scFV engineered biomimetic nanovesicles to overcome BC in primary, recurrent, metastatic and abscopal humanized mice models. Nanovesicles coated anti-CD276 scFV selectively fuses with cell membrane of senescent-escape CSCs, then sequentially delivers siCALD1 and ADR due to pH-responsive MnP shell. siCALD1 together with ADR effectively induce apoptosis of CSCs, decrease expression of CD276 and PD-L1, and upregulate MHC I combined with Mn2+ to overcome chemoresistance and promote CD8+T cells infiltration. This combined therapeutic approach reveals insights into immune surveillance evasion by senescent-escape CSCs, offering a promising strategy to immunotherapy effectiveness in cancer therapy.

Key traits influencing the resistance of Eucalyptus camaldulensis to wind damage in coastal areas of South China.

Aims: China is one of the countries in the world most seriously affected by typhoons, which pose a great threat to the eucalyptus plantation industry. However, few studies have comprehensively accounted for the impact of key traits on the wind damage/resistance of eucalyptus. Methods: To identify the key factors affecting the wind resistance of eucalyptus, 20 eucalyptus genotypes were selected; a total of 18 traits, including the wind damage index, growth traits, and wood traits, were measured, and the wind resistance was determined via the tree-pulling test. Results: Correlation, principal component, canonical correlation, and path analyses were performed to evaluate these traits. Correlation analysis revealed that the wind resistance of eucalyptus plants was related to the tree height, volume, and duration of stress wave propagation. Principal components and tree-pulling variables were further used for correlation and path analyses. Canonical correlation analysis and the PA-OV model showed that holocellulose and lignin contents and fiber width, as well as growth traits, were important factors affecting the stability of standing trees under typhoon conditions. The key traits influencing the wind resistance of Eucalyptus camaldulensis, which may provide a reference for evaluating the wind resistance of Eucalyptus varieties for forest management, were identified. Conclusion: This study provides a knowledge base for forest management and planning in typhoon-prone coastal areas, and provides a theoretical basis for the breeding and genetically improving eucalyptus stocks based on wind resistance characteristics.

Correction: Characterization of two constitutive promoters RPS28 and EIF1 for studying soybean growth, development, and symbiotic nodule development.

[This corrects the article DOI: 10.1007/s42994-022-00073-6.].

The Chrysosplenium sinicum genome provides insights into adaptive evolution of shade plants.

Chrysosplenium sinicum, a traditional Tibetan medicinal plant, can successfully thrive in low-light environments for long periods of time. To investigate the adaptive evolution of shade plants in low-light environments, we generated a chromosome-scale genome assembly (~320 Mb) for C. sinicum by combining PacBio sequencing and Hi-C technologies. Based on our results, gene families related to photosynthesis and cell respiration greatly expanded and evolved in C. sinicum genome due to intracellular DNA transfer from organelle genome to nuclear genome. Under positive selective pressure, adaptive evolution of light-harvesting complex II (LHCII) component protein CsLhcb1s resulted in the expansion of threonine residues at the phosphorylation site of STN7 kinase, potentially establishing a crucial genomic foundation for enhancing C. sinicum's adaptability in low-light environments. Through transcriptome and metabolome analysis, we identified chrysosplenol and chrysosplenoside as predominant flavonoid metabolites of C. sinicum and predicted their synthesis pathways. In addition, analysis of alternative splicing (AS) revealed that AS events help regulate state transition and flavonoid biosynthesis. The present study provides new insights into the genomes of shade plants exposed to low-light conditions and adaptive evolution of these genomes; in addition, the results improve our current knowledge on the biosynthetic and regulatory processes of chrysosplenol and chrysosplenoside.

The enhanced neutral process with decreasing cell size: a study on phytoplankton metacommunities from the glacier-fed river of Qinghai-Xizang Plateau.

The cell size of phytoplankton is an important defining functional trait that can serve as a driver and sentinel of phytoplankton community structure and function. However, the study of the assembly patterns and drivers of phytoplankton metacommunities with different cell sizes has not been widely carried out. In this study, we systematically investigated the biodiversity patterns, drivers, and assembly processes of the three phytoplankton cell sizes (micro: 20-200 µm; nano: 2-20 µm; pico: 0.2-2 µm) in the Za'gya Zangbo River from the source to the estuary using 18S rDNA amplicon sequencing. The results demonstrated that the alpha diversity and co-occurrence network complexity for all three sizes of phytoplankton increased to a peak downstream of the glacier sources and then decreased to the estuary. The nanophytoplankton subcommunity consistently had the highest alpha diversity and co-occurrence network complexity. On the other hand, total beta diversity followed a unimodal trend of decreasing and then increasing from source to estuary, and was dominated by species replacement components. In addition, deterministic processes driven mainly by physiochemical indices (PCIs) and biogenic elements (BGEs) dominated the assembly of micro- and nanophytoplankton subcommunities, whereas stochastic processes driven by geographical factors (GGFs) dominated the assembly of picophytoplankton subcommunities. The results explained the contradictions in previous studies of phytoplankton community assembly processes in highland aquatic ecosystems, elucidating the different contributions of deterministic and stochastic processes, and the complexity of compositional mechanisms in shaping the assembly of micro-, nano-, and picophytoplankton in this highland glacial river. IMPORTANCE: The cell size of phytoplankton is a key life-history trait and key determinant, and phytoplankton of different cell sizes are differentially affected by ecological processes. However, the study of the assembly patterns and drivers of phytoplankton metacommunities with different cell sizes has not been widely carried out. We provide an in-depth analysis of phytoplankton community diversity across three cell sizes in the glacier-fed river, describing how the pattern of phytoplankton communities differs across cell sizes in response to geochemical gradients. The results show that the smaller phytoplankton (picophytoplankton) are relatively more influenced by dispersal-based stochastic processes, whereas larger ones (microphytoplankton and nanophytoplankton) are more structured by selection-based deterministic processes.

A composite enzyme derived from papain and chymotrypsin reduces the Allergenicity of Cow's Milk allergen casein by targeting T and B cell epitopes.

Casein, the major allergen in cow's milk, presents a significant challenge in providing nutritional support for children with allergies. To address this issue, we investigated a composite enzyme, comprising papain and chymotrypsin, to reduce the allergenicity of casein. Enzymatic hydrolysis induced substantial structural changes in casein, diminishing its affinity for specific IgE and IgG antibodies. Additionally, in a BALB/c mouse model, casein hydrolysate alleviated allergic symptoms, evidenced by lower serum IgE and IgG levels, reduced plasma histamine, and decreased Th2 cytokine release during cell co-culture. Peptidomic analysis revealed a 52.38% and 60% reduction in peptides containing IgE epitopes in casein hydrolyzed by the composite enzyme compared to papain and chymotrypsin, respectively, along with a notable absence of previously reported T cell epitopes. These results demonstrate the potential of enzyme combinations to enhance the efficiency of epitope destruction in allergenic proteins, providing valuable insights into the development of hypoallergenic dairy products.

Moving object detection based on bioinspired background subtraction.

Flying insects rely mainly upon visual motion to detect and track objects. There has been a lot of research on fly inspired algorithms for object detection, but few have been developed based on visual motion alone. One of the daunting difficulties is that the neural and circuit mechanisms underlying the foreground-background segmentation are still unclear. Our previous modeling study proposed that the lobula held parallel pathways with distinct directional selectivity, each of which could retinotopically discriminate figures moving in its own preferred direction based on relative motion cues. The previous model, however, did not address how the multiple parallel pathways gave the only detection output at their common downstream. Since the preferred directions of the pathways along either horizontal or vertical axis were opposite to each other, the background moving in the opposite direction to an object also activated the corresponding lobula pathway. Indiscriminate or ungated projection from all the pathways to their downstream would mix objects with the moving background, making the previous model fail with non-stationary background. Here, we extend the previous model by proposing that the background motion-dependent gating of individual lobula projections is the key to object detection. Large-field lobula plate tangential cells are hypothesized to perform the gating to realize bioinspired background subtraction. The model is shown to be capable of implementing a robust detection of moving objects in video sequences with either a moving camera that induces translational optic flow or a static camera. The model sheds light on the potential of the concise fly algorithm in real-world applications.

Sprayable, thermosensitive hydrogels for promoting wound healing based on hollow, porous and pH-sensitive ZnO microspheres.

A solvothermal method and the subsequent heat treatment process were developed to fabricate hollow ZnO particles with hierarchical pores on a large scale. The as-obtained hollow, porous ZnO microspheres with tunable sizes, high specific surface areas, pH sensitivity, antibacterial properties, and high adsorption capacities showed significant advantages for drug delivery. Sprayable hydrogels containing hollow, porous ZnO microspheres and curcumin nanoparticles (CNPs) were prepared to accelerate wound healing. The water-dispersed CNPs promoted both the migration of fibroblasts and angiogenesis and an aqueous solution of Pluronic F127 (a temperature-sensitive phase-change hydrogel material) was shown to be an effective choice for medical dressings. The experimental data suggest that hollow, porous ZnO microspheres can be loaded with additional CNPs to achieve continuous long-term therapeutic effects.

A Comparative Study of a Nomogram and Machine Learning Models in Predicting Early Hematoma Expansion in Hypertensive Intracerebral Hemorrhage.

RATIONALE AND OBJECTIVES: Early identification for hematoma expansion can help improve patient outcomes. Presently, there are many methods to predict hematoma expansion. This study compared a variety of models to find a model suitable for clinical promotion. MATERIALS AND METHODS: Non-contrast head CT images and clinical data were collected from 203 patients diagnosed with hypertensive intracerebral hemorrhage. Radiomics features were extracted from all CT images, and the dataset was randomly divided into training and validation sets (7:3 ratio) after applying the synthetic minority oversampling method. The radiomics score (Radscore) was calculated using least absolute shrinkage and selection operator (LASSO) regression, combined with selected clinical predictors, to develop a nomogram and four machine learning (ML) models: logistic regression, random forest, support vector machine, and extreme gradient boosting (XGBoost). Discrimination, calibration and clinical usefulness of the nomogram and ML models were assessed. RESULTS: The nomogram and ML models were integrated with Radscore and clinical predictors. The nomogram demonstrated favorable discriminatory ability in the training set with an AUC of 0.80, which was confirmed in the validation set (AUC=0.76). Among the ML models, the XGBoost model achieved the highest AUC (training set=0.89 and validation set=0.85), surpassing that of the nomogram. The XGBoost model exhibited good clinical usefulness. CONCLUSION: Both the nomogram and ML models constructed by non-contrast head CT image-based Radscore integrated with clinical predictors can predict early hematoma expansion of hypertensive intracerebral hemorrhage, and the XGBoost model had the highest prediction performance and best clinical usefulness.

Mitochondria targeted drug delivery system overcoming drug resistance in intrahepatic cholangiocarcinoma by reprogramming lipid metabolism.

The challenge of drug resistance in intrahepatic cholangiocarcinoma (ICC) is intricately linked with lipid metabolism reprogramming. The hepatic lipase (HL) and the membrane receptor CD36 are overexpressed in BGJ398-resistant ICC cells, while they are essential for lipid uptake, further enhancing lipid utilization in ICC. Herein, a metal-organic framework-based drug delivery system (OB@D-pMOF/CaP-AC, DDS), has been developed. The specifically designed DDS exhibits a successive targeting property, enabling it to precisely target ICC cells and their mitochondria. By specifically targeting the mitochondria, DDS produces reactive oxygen species (ROS) through its sonodynamic therapy effect, achieving a more potent reduction in ATP levels compared to non-targeted approaches, through the impairment of mitochondrial function. Additionally, the DDS strategically minimizes lipid uptake through the incorporation of the anti-HL drug, Orlistat, and anti-CD36 monoclonal antibody, reducing lipid-derived energy production. This dual-action strategy on both mitochondria and lipids can hinder energy utilization to restore drug sensitivity to BGJ398 in ICC. Moreover, an orthotopic mice model of drug-resistant ICC was developed, which serves as an exacting platform for evaluating the multifunction of designed DDS. Upon in vivo experiments with this model, the DDS demonstrated exceptional capabilities in suppressing tumor growth, reprogramming lipid metabolism and improving immune response, thereby overcoming drug resistance. These findings underscore the mitochondria-targeted DDS as a promising and innovative solution in ICC drug resistance.

Ovalbumin promotes innate immune response of Caenorhabditis elegans through DAF-16 and SKN-1 pathways in insulin/IGF-1 signaling.

Ovalbumin (OVA) is a major allergen in eggs and could induce severe allergic reactions in sensitive individuals, where the innate immune system works as a regulator. The mechanism of how innate immunity adjusts to food allergy is relatively well-studied, however, the effects of allergen uptake on the innate immune system remain unclear. Therefore, the Caenorhabditis elegans (C. elegans) model was utilized to assess the effects of OVA on its innate immune system. OVA enhanced the immune response of C. elegans with higher survival rates under Pseudomonas aeruginosa infection. Moreover, sustaining OVA treatment improved the health states that were reflected in the prolonged lifespan, alleviated oxidative stress, accelerated growth, and promoted motility. RNA-sequencing analysis and the slow-killing assays in the mutants of insulin/IGF-1 signaling (IIS)-related genes confirmed that IIS was necessary for OVA to regulate innate immunity. Besides, OVA activated SKN-1 temporarily and facilitated the nuclear localization of DAF-16 for improving immunity and health status in C. elegans. Together, OVA could enhance the innate immune responses via DAF-16 and SKN-1 pathways in the IIS of C. elegans, and this work will provide novel insights into the regulation of innate immunity by OVA in higher organisms.

Diversity Patterns of Eukaryotic Phytoplankton in the Medog Section of the Yarlung Zangbo River.

As one of the important biodiversity conservation areas in China, the ecosystem in the lower reaches of the Yarlung Zangbo River is fragile, and is particularly sensitive to global changes. To reveal the diversity pattern of phytoplankton, the metabarcode sequencing was employed in the Medog section of the lower reaches of the Yarlung Zangbo River during autumn 2019 in present study. The phytoplankton assemblies can be significantly divided into the main stem and the tributaries; there are significant differences in the phytoplankton biomass, alpha and beta diversity between the main stem and the tributaries. While both the main stem and the tributaries are affected by dispersal limitation, the phytoplankton assemblages in the entire lower reaches are primarily influenced by heterogeneous selection. Community dissimilarity and assembly process were significantly correlated with turbidity, electrical conductivity, and nitrogen nutrition. The tributaries were the main source of the increase in phytoplankton diversity in the lower reaches of the Yarlung Zangbo River. Such diversity pattern of phytoplankton in the lower reach may be caused by the special habitat in Medog, that is, the excessive flow velocity, and the significant spatial heterogeneity in physical and chemical factors between stem and tributaries. Based on the results and conclusions obtained in present study, continuous long-term monitoring is essential to assess and quantify the impact of global changes on phytoplankton.

Transglutaminase-Cross-Linked Tofu Suppressed Soybean-Induced Allergic Reactions by Enhancing Intestinal Mucosa Immune Tolerance.

Currently, food allergies are closely related to intestinal health, and ensuring the integrity and health of intestinal mucosa could reduce the incidence of food allergies. In this study, a soybean-allergic mouse model was used to explore the mechanism of intestinal mucosa immune response induced by enzyme-cross-linked tofu. The effects of enzyme-cross-linked tofu on intestinal mucosal immunity in mice were determined by hematoxylin-eosin (HE) staining and flow cytometry. Our results reveled that the MTG-cross-linked tofu reduced the reactivity of the intestinal mucosal immune system, which mainly manifested as a decrease in the dendritic cell (DC) levels of mesenteric lymph nodes (MLNs), increasing the Th1 cells and Tregs in Peyer's patch (PP) nodes and MLNs, and inhibiting the Th2 cells. Compared with soy protein, enzyme-cross-linked tofu had less damage to the small intestinal tract of mice. Therefore, the above-mentioned results fully revealed that the enzyme-cross-linked tofu promoted the transformation of intestinal mucosal immune cells, shifted the Th1/Th2 balance toward Th1, and reduced its sensitization effect.

The chromosome-scale genome and population genomics reveal the adaptative evolution of Populus pruinosa to desertification environment.

The Populus pruinosa is a relic plant that has managed to survive in extremely harsh desert environments. Owing to intensifying global warming and desertification, research into ecological adaptation and speciation of P. pruinosa has attracted considerable interest, but the lack of a chromosome-scale genome has limited adaptive evolution research. Here, a 521.09 Mb chromosome-level reference genome of P. pruinosa was reported. Genome evolution and comparative genomic analysis revealed that tandemly duplicated genes and expanded gene families in P. pruinosa contributed to adaptability to extreme desert environments (especially high salinity and drought). The long terminal repeat retrotransposons (LTR-RTs) inserted genes in the gene body region might drive the adaptive evolution of P. pruinosa and species differentiation in saline-alkali desert environments. We recovered genetic differentiation in the populations of the northern Tianshan Mountain and southern Tianshan Mountain through whole-genome resequencing of 156 P. pruinosa individuals from 25 populations in China. Further analyses revealed that precipitation drove the local adaptation of P. pruinosa populations via some genetic sites, such as MAG2-interacting protein 2 (MIP2) and SET domain protein 25 (SDG25). This study will provide broad implications for adaptative evolution and population studies by integrating internal genetic and external environmental factors in P. pruinosa.

Assessment of sepsis-associated encephalopathy by quantitative magnetic resonance spectroscopy in a rat model of cecal ligation and puncture.

Proton magnetic resonance spectroscopy (1H-MRS) is the only non-invasive technique to quantify neurometabolic compounds in the living brain. We used 1H-MRS to evaluate the brain metabolites in a rat model of Sepsis-associated encephalopathy (SAE) established by cecal ligation and puncture (CLP). 36 male Sprague-Dawley rats were randomly divided into sham and CLP groups. Each group was further divided into three subgroups: subgroup O, subgroup M, and subgroup N. Neurological function assessments were performed on the animals in the subgroup O and subgroup N at 24 h, 48 h, and 72 h. The animals in the subgroup M were examined by magnetic resonance imaging (MRI) at 12 h after CLP. Compared with the sham group, the ratio of N-acetylaspartate (NAA) to creatine (Cr) in the hippocampus was significantly lower in the CLP group. The respective ratios of lactate (Lac), myo-inositol (mIns), glutamate and glutamine (Glx), lipid (Lip), and choline (Cho) to Cr in the CLP group were clearly higher than those in the sham group. Cytochrome c, intimately related to oxidative stress, was elevated in the CLP group. Neurofilament light (NfL) chain and glial fibrillary acidic protein (GFAP) scores in the CLP group were significantly higher than those in the sham group, while zonula occludens-1 (ZO-1) was downregulated. Compared with the sham group, the CLP group displayed higher values of oxygen extraction fraction (OEF), central venous-arterial partial pressure of carbon dioxide (P (cv-a) CO2), and central venous lactate (VLac). In contrast, jugular venous oxygen saturation (SjvO2) declined. In the present study, 1H-MRS could be used to quantitatively assess brain injury in terms of microcirculation disorder, oxidative stress, blood-brain barrier disruption, and glial cell activation through changes in metabolites within brain tissue.

Nanosystem Delivers Senescence Activators and Immunomodulators to Combat Liver Cancer.

CD47 blockade has emerged as a promising immunotherapy against liver cancer. However, the optimization of its antitumor effectiveness using efficient drug delivery systems or combinations of therapeutic agents remains largely incomplete. Here, patients with liver cancer co-expressing CD47 and CDC7 (cell division cycle 7, a negative senescence-related gene) are found to have the worst prognosis. Moreover, CD47 is highly expressed, and senescence is inhibited after the development of chemoresistance, suggesting that combination therapy targeting CD47 and CDC7 to inhibit CD47 and induce senescence may be a promising strategy for liver cancer. The efficacy of intravenously administered CDC7 and CD47 inhibitors is limited by low uptake and short circulation times. Here, inhibitors are coloaded into a dual-targeted nanosystem. The sequential release of the inhibitors from the nanosystem under acidic conditions first induces cellular senescence and then promotes immune responses. In an in situ liver cancer mouse model and a chemotherapy-resistant mouse model, the nanosystem effectively inhibited tumor growth by 90.33% and 85.15%, respectively. Overall, the nanosystem in this work achieved the sequential release of CDC7 and CD47 inhibitors in situ to trigger senescence and induce immunotherapy, effectively combating liver cancer and overcoming chemoresistance.

Anthropogenic impacts and quantitative sources of nitrate in a rural-urban canal using a combined PMF, δ15N/δ18O-NO3-, and MixSIAR approach.

Nitrate (NO3-) pollution in irrigation canals is of great concern because it threatens canal water use; however, little is known about it at present. Herein, a combination of positive matrix factorization (PMF), isotope tracers, and Mixing Stable Isotope Analysis in R (MixSIAR) was developed to identify anthropogenic impacts and quantitative sources of NO3- in a rural-urban canal in China. The NO3- concentration (0.99-1.93 mg/L) of canal water increased along the flow direction and was higher than the internationally recognized eutrophication risk value in autumn and spring. The inputs of the Fuhe River, NH4+ fertilizer, soil nitrogen, manure & sewage, and rainfall were the main driving factors of canal water NO3- based on principal component analysis and PMF, which was supported by evidence from δ15N/δ18O-NO3-. According to the chemical and isotopic analyses, nitrogen transformation was weak, highlighting the potential of δ15N/δ18O-NO3- to trace NO3- sources in canal water. The MixSIAR and PMF results with a <15% divergence emphasized the predominance of the Fuhe River (contributing >50%) and anthropogenic impacts (NH4+ fertilizer plus manure & sewage, >37%) on NO3- in the entire canal, reflecting the effectiveness of the model analysis. According to the MixSIAR model, (1) higher NO3- concentration in canal water was caused by the general enhancement of human activities in spring and (2) NO3- source contributions were associated with land-use patterns. The high contributions of NH4+ fertilizer and manure & sewage showed inverse spatial variations, suggesting the necessity of reducing excessive fertilizer use in the agricultural area and controlling blind wastewater release in the urban area. These findings provide valuable insights into NO3- dynamics and fate for sustainable management of canal water resources. Nevertheless, long-term chemical and isotopic monitoring with alternative modeling should be strengthened for the accurate evaluation of canal NO3- pollution in future studies.

Comparative genomic analysis of the Growth-Regulating Factors-Interacting Factors (GIFs) in six Salicaceae species and functional analysis of PeGIF3 reveals their regulatory role in Populus heteromorphic leaves.

BACKGROUND: The growth-regulating factor-interacting factor (GIF) gene family plays a vital role in regulating plant growth and development, particularly in controlling leaf, seed, and root meristem homeostasis. However, the regulatory mechanism of heteromorphic leaves by GIF genes in Populus euphratica as an important adaptative trait of heteromorphic leaves in response to desert environment remains unknown. RESULTS: This study aimed to identify and characterize the GIF genes in P. euphratica and other five Salicaceae species to investigate their role in regulating heteromorphic leaf development. A total of 27 GIF genes were identified and characterized across six Salicaceae species (P. euphratica, Populus pruinose, Populus deltoides, Populus trichocarpa, Salix sinopurpurea, and Salix suchowensis) at the genome-wide level. Comparative genomic analysis among these species suggested that the expansion of GIFs may be derived from the specific Salicaceae whole-genome duplication event after their divergence from Arabidopsis thaliana. Furthermore, the expression data of PeGIFs in heteromorphic leaves, combined with functional information on GIF genes in Arabidopsis, indicated the role of PeGIFs in regulating the leaf development of P. euphratica, especially PeGIFs containing several cis-acting elements associated with plant growth and development. By heterologous expression of the PeGIF3 gene in wild-type plants (Col-0) and atgif1 mutant of A. thaliana, a significant difference in leaf expansion along the medial-lateral axis, and an increased number of leaf cells, were observed between the overexpressed plants and the wild type. CONCLUSION: PeGIF3 enhances leaf cell proliferation, thereby resulting in the expansion of the central-lateral region of the leaf. The findings not only provide global insights into the evolutionary features of Salicaceae GIFs but also reveal the regulatory mechanism of PeGIF3 in heteromorphic leaves of P. euphratica.

Mass Spectrometry Analysis on the Breakage of Allergens in High-Molecular-Mass Polymer of Roasted Peanuts.

Peanut allergy is a prevalent and concerning food allergy. Roasting can introduce structural changes to peanut allergens, affecting their allergenicity, but the structure on the primary structure is unclear. Here, the breakage sites were identified by mass spectrometry and software tools, and structural changes were simulated by molecular dynamics and displayed by PyMOL software. Results revealed that the appearance frequencies of L, Q, F, and E were high at the N-terminal of the breakage site, while S and E were dominant at the C-terminal. In the conformational structure, breakage sites were found close to disulfide bonds and the Cupin domains of Ara h 1 and Ara h 3. The breakage of allergens destroyed linear epitopes and might change the conformation of epitopes, which could influence peanuts' potential allergenicity.

The structure and potential allergenicity of peanut allergen monomers after roasting.

Given that roasted peanut (Ro) products are commonly used in daily life, peanut allergenicity is a foremost concern. Analyzing the changes in the structure and potential allergenicity of individual allergens can promote the exploration of the structural basis of the alterations in the potential allergenicity of Ro. This work focused on four major allergens in raw peanut (Ra) and Ro. Structural changes were analyzed on the basis of circular dichroism, ultraviolet and fluorescence spectroscopy, and molecular dynamic simulation. The IgE recognition capability of allergens was assessed via western blot analysis. The IgE binding capacity of allergens was detected by conducting enzyme-linked immunosorbent assay. The potential allergenicity of allergens was evaluated using the KU812 cell degranulation model. The results showed that roasting induced different changes in the overall structures of allergens and altered the structures and electrostatic potential of IgE epitopes, especially Ara h 1 and Ara h 6. These alterations affected the potential allergenicity of allergens. Ara h 1 and Ara h 6 in Ro showed significantly enhanced IgE binding capacities and abilities to elicit KU812 cell degranulation, while Ara h 2 and Ara h 3 did not change significantly. For total protein, the roasted peanut protein showed decreased abilities to elicit KU812 cell degranulation. The results indicated that different allergens in Ro showed different changes of structures and potential allergenicity and that the conformational structure plays a crucial role in potential allergenicity of allergens.