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Abstract Broadly neutralizing antibodies (bNAbs) could offer escape-tolerant and lasting protection against viral infections and therefore guide development of broad-spectrum vaccines. The increasing challenge posed by viral evolution and immune evasion intensifies the importance of the discovery of bNAbs and their underlying neutralization mechanism. Here, focusing on the pivotal viral protein VP4 of rotavirus (RV), we identify a potent bNAb, 7H13, exhibiting broad-spectrum neutralization across diverse RV genotypes and demonstrating strong prevention of virus infection in female mice. A combination of time-resolved cryo-electron microscopy (cryo-EM) and in situ cryo-electron tomography (cryo-ET) analysis reveals a counterintuitive dynamic process of virus inactivation, in which 7H13 asymmetrically binds to a conserved epitope in the capsid-proximal aspect of VP4, triggers a conformational switch in a critical residue—F418—thereby disrupts the meta-stable conformation of VP4 essential for normal viral infection. Structure-guided mutagenesis corroborates the essential role of the 7H13 heavy chain I54 in activating F418 switch and destabilizing VP4. These findings define an atypical NAbs’ neutralization mechanism and reveal a potential type of virus vulnerable site for universal vaccine and therapeutics design.

Abstract The numerous high-risk carcinogenic types of human papillomavirus (HR-HPV) that lack vaccine protection underscore the urgent need to develop broader-spectrum HPV vaccines. This study addresses this need by focusing on HR-HPV types 53, 56, and 66, which are not currently targeted by existing vaccines. It introduces an effective method for their soluble expression, as well as that of their mutants, within an Escherichia coli expression system. Through strategic homologous loop swapping among HPV53, HPV56, and HPV66, we designed twenty double-type chimeric molecules. Comprehensive evaluations identified unique dominant immunogenic loops for each type: the FG loop for HPV53, the HI loop for HPV56, and the DE loop for HPV66, with HPV66 emerging as the optimal chimeric backbone virus-like particle (VLP). By incorporating two identified immunodominant sites into the preferable base particle, the study constructed a triple-type chimera H66-56HI-53FG, which could efficiently self-assemble into VLPs in vitro that closely resembled the wild-type HPV66 VLP and, induced balanced triple-type neutralization titers (~ 3 log unites), as contrast to none observable HPV53 neutralization titer and lower HPV56 titer elicited by the immunization of the wild-type HPV66 alone. This research outlines an amenable way to simultaneously identify immunodominant sites and their preferable particle base context for cross-type vaccine design, thereby offering a paradigm as extending antigenic variety in single particle to broaden vaccine protection coverage. Graphical Abstract

Abstract Background Targeting the TGF-β pathway in tumor therapy has proven challenging due to the highly context-dependent functions of TGF-β. Integrin αvβ8, a pivotal activator of TGF-β, has been implicated in TGF-β signaling within tumors, as demonstrated by the significant anti-tumor effects of anti-αvβ8 antibodies. Nevertheless, the expression profile of αvβ8 remains a subject of debate, and the precise mechanisms underlying the anti-tumor effects of anti-αvβ8 antibodies are not yet fully elucidated. Methods We utilized single-cell RNA sequencing to assess αvβ8 expression across various human tumors. An anti-αvβ8 antibody was developed and characterized for its binding and blocking properties in vitro. Cryo-EM single-particle analysis was employed to study the detailed interaction between αvβ8 and the antibody Fab fragment. The anti-tumor efficacy of the antibody was evaluated in syngeneic mouse models with varying levels of αvβ8 expression, both as a monotherapy and in combination with PD-1 antibodies. Human PBMCs were isolated to investigate αvβ8 expression in myeloid cells, and macrophages were exposed to the antibody to study its impact on macrophage polarization. Pharmacokinetic studies of the αvβ8 antibody were conducted in cynomolgus monkeys. Results Integrin αvβ8 is notably expressed in certain tumor types and tumor-infiltrating macrophages. The specific αvβ8 antibody 130H2 demonstrated high affinity, specificity, and blocking potency in vitro. Cryo-EM analysis further revealed that 130H2 interacts exclusively with the β8 subunit, without binding to the αv subunit. In vivo studies showed that this antibody significantly inhibited tumor growth and alleviated immunosuppression by promoting immune cell infiltration. Furthermore, combining the antibody with PD-1 inhibition produced a synergistic anti-tumor effect. In human PBMCs, monocytes exhibited high αvβ8 expression, and the antibody directly modulated macrophage polarization. Tumors with elevated αvβ8 expression were particularly responsive to 130H2 treatment. Additionally, favorable pharmacokinetic properties were observed in cynomolgus monkeys. Conclusions In summary, integrin αvβ8 is highly expressed in certain tumors and tumor-infiltrating macrophages. Targeting αvβ8 with a blocking antibody significantly inhibits tumor growth by modulating macrophage polarization and enhancing immune cell infiltration. Combining αvβ8 targeting with PD-1 treatment markedly increases the sensitivity of immune-excluded tumors. These results support further clinical evaluation of αvβ8 antibodies. Graphical abstract

Abstract Respiratory syncytial virus (RSV) poses a significant public health challenge, especially among children. Although palivizumab and nirsevimab, neutralizing antibodies (nAbs) targeting the RSV F protein, have been used for prophylaxis, their limitations underscore the need for more effective alternatives. Herein, we present a potent and broad nAb, named 5B11, which exhibits nanogram level of unbiased neutralizing activities against both RSV-A and -B subgroups. Notably, 5B11 shows a ~20-fold increase in neutralizing efficacy compared to 1129 (the murine precursor of palivizumab) and approximately a 3-fold increase in neutralizing efficacy against B18537 in comparison to nirsevimab. Cryo-electron microscopy analysis reveals 5B11’s mechanism of action by targeting a highly conserved epitope within site V, offering a promising strategy with potentially lower risk of escape mutants. Antiviral testing in a female cotton rat model demonstrated that low-dose (1.5 mg/kg) administration of 5B11 achieved comparable prophylactic efficacy to that achieved by high-dose (15 mg/kg) of 1129. Furthermore, the humanized 5B11 showed a superior in vivo antiviral activity against B18537 infection compared to nirsevimab and palivizumab. Therefore, 5B11 is a promising RSV prophylactic candidate applicable to broad prevention of RSV infection.

Optogenetics and chemogenetics are emerging neuromodulation techniques that have attracted significant attention in recent years. These techniques enable the precise control of specific neuronal types and neural circuits, allowing researchers to investigate the cellular mechanisms underlying depression. The advancement in these techniques has significantly contributed to the understanding of the neural circuits involved in depression; when combined with other emerging technologies, they provide novel therapeutic targets and diagnostic tools for the clinical treatment of depression. Additionally, these techniques have provided theoretical support for the development of novel antidepressants. This review primarily focuses on the application of optogenetics and chemogenetics in several brain regions closely associated with depressive-like behaviors in rodent models, such as the ventral tegmental area, nucleus accumbens, prefrontal cortex, hippocampus, dorsal raphe nucleus, and lateral habenula and discusses the potential and challenges of optogenetics and chemogenetics in future research. Furthermore, this review discusses the potential and challenges these techniques pose for future research and describes the current state of research on sonogenetics and odourgenetics developed based on optogenetics and chemogenetics. Specifically, this study aimed to provide reliable insights and directions for future research on the role of optogenetics and chemogenetics in the neural circuits of depressive rodent models.

Background Patients with microsatellite stable (MSS) colorectal cancer (CRC) often display resistance to immunotherapy. Epidermal growth factor receptor (EGFR)-targeted therapies have shown potential in enhancing immunotherapy, yet clinical benefits remain unfulfilled, which may relate to inadequate patient stratification.Methods Circulating tumor cells and tumor tissues were collected from multicenter cohorts of patients with CRC receiving cetuximab to analyze EGFR variant type III (EGFRvIII) expression and immune infiltration. Syngeneic mouse models of EGFRvIII CRC were used to investigate the combined efficacy of adenosine inhibition and antiprogrammed cell death protein 1 (anti-PD-1).Results EGFRvIII mutations are found in about 10% of MSS CRC and are associated with poor response to cetuximab therapy. EGFRvIII-mutated patients with CRC exhibit an adenosine-mediated immunosuppressive tumor microenvironment (TME) subtype. Combination therapy with adenosine inhibitors remodels the TME, reversing cetuximab resistance and enhancing anti-PD-1 efficacy in EGFRvIII CRC.Conclusions Our findings identified EGFRvIII-positive CRC as a distinct subtype characterized by adenosine-mediated immunosuppressive TME. Targeting adenosine significantly improved the efficacy of anti-PD-1 in MSS CRC.

Abstract Background An increasing number of clinical studies have begun to explore combination strategies with immune checkpoint inhibitors, aiming to present new opportunities for overcoming anti-PD-1 treatment resistance in gastric cancer. Unfortunately, the exploration of certain immune checkpoint inhibitor combination strategies has yielded suboptimal results. Therefore, it is necessary to comprehensively analyze the expression patterns of immune checkpoints and identify optimal combination regimens of anti-PD-1 inhibitors with other immune checkpoint inhibitors. Methods Leveraging single-cell RNA sequencing (scRNA-seq) and multivariate linear regression interaction models, we dissected the immune checkpoint expression characteristics of CD8+ T cells in gastric cancer and the immune checkpoint expression pattern (ICEP) mediating anti-PD-1 treatment resistance. Furthermore, we employed transcription factor analysis and CellOracle to explore the transcriptional regulatory mechanisms governing CD8+ T cell differentiation fates. Finally, we utilized Nichenet and spatial transcriptomic analysis to investigate the spatial expression patterns of immune checkpoints. Results Interaction analysis indicated that, among the known immune checkpoints, co-expression of NKG2A and PD-1 might exert a more profound inhibitory effect on the proliferative capacity of CD8+ T cells. The co-expression analysis revealed differential co-expression pattern of PD-1 and NKG2A, defined as ICEP1 (CD8+ T cells co-expressing PD-1, CTLA-4, TIGIT, LAG-3 or CD38) and ICEP2 (CD8+ T cells solely expressing NKG2A or co-expressing with other immune checkpoints), reflecting the co-occurrence pattern of PD-1 and the mutual exclusivity of NKG2A. Further, these two ICEP CD8+ T cell subsets represented distinct CD8+ T cell differentiation fates governed by MSC and RUNX3. Notably, ICEP2 CD8+ T cells were associated with anti-PD-1 therapy resistance in gastric cancer. This phenomenon may be attributed to the recruitment of LGMN+ macrophages mediated by the CXCL16-CXCR6 signaling pathway. Conclusion This study unveiled two distinct ICEPs and the mutually exclusivity and co-occurrence characteristics of CD8+ T cells in gastric cancer. The ICEP2 CD8+ T cell subset, highly expressed in gastric cancer patients resistant to anti-PD-1 therapy, may be recruited by LGMN+ macrophages through CXCL16-CXCR6 axis. These findings provide evidence for NKG2A as a novel immunotherapeutic target in gastric cancer and offer new insights into combination strategies for immune checkpoint inhibitors in gastric cancer. Graphical Abstract

Abstract Timely and effective diagnosis of fungal keratitis (FK) is necessary for suitable treatment and avoiding irreversible vision loss for patients. In vivo confocal microscopy (IVCM) has been widely adopted to guide the FK diagnosis. We present a deep learning framework for diagnosing fungal keratitis using IVCM images to assist ophthalmologists. Inspired by the real diagnostic process, our method employs a two-stage deep architecture for diagnostic predictions based on both image-level and sequence-level information. To the best of our knowledge, we collected the largest dataset with 96,632 IVCM images in total with expert labeling to train and evaluate our method. The specificity and sensitivity of our method in diagnosing FK on the unseen test set achieved 96.65% and 97.57%, comparable or better than experienced ophthalmologists. The network can provide image-level, sequence-level and patient-level diagnostic suggestions to physicians. The results show great promise for assisting ophthalmologists in FK diagnosis.

Abstract The Mu Us Sandy Land is a region characterized by wind-blown sand and soil erosion in northern China. To enhance the soil quality of this area, various organic materials were incorporated into the mixed soil at a volume ratio of 1:2 for feldspathic sandstone to sand. Culture was conducted in the field and under constant temperature conditions in laboratory culture chambers. Four treatments were established in the experiment, each calculated based on weight ratio and controlled (with no organic material added, CK); single application of straw (5% straw, P1); single application of biochar (5% biochar, P2); combined application of biochar and straw (5% biochar + 5% straw, P3). After 90 days of culture, soil samples were collected for analysis of various indicators such as soil aggregate particle size distribution, water stability of soil aggregates, mean weight diameter, mean geometric diameter, and fractal dimension using dry sieving and wet sieving methods. The objective is to establish a scientific basis and provide technical support for addressing the challenges associated with compound soil and implementing rational fertilization measures. The research results indicate that: (1) The quantity of aggregates > 0.25 mm under different treatments follows the order CK 0.25 (x) of compound soil, with a correlation coefficient of up to 0.9851. In conclusion, the incorporation of organic materials can effectively enhance the proportion of macroaggregates in compound soil consisting of Feldspathic sandstone and sand, thereby improving soil stability and erosion resistance. The optimal outcome is achieved through the combined application of biochar and straw. Indoor culture proves to be more effective than field culture, while wet sieving accurately reflects the structural characteristics of compound soil under both dry and wet sieving treatments.

Abstract Cellulosic materials are attracting increasing research interest because of their abundance, biocompatibility, and biodegradability, making them suitable in multiple industrial and medical applications. Functionalization of cellulose is usually required to improve or expand its properties to meet the requirements of different applications. Cellulose-binding domains (CBDs) found in various proteins have been shown to be powerful tools in the functionalization of cellulose materials. In this review, we firstly introduce the structural characteristics of commonly used CBDs belonging to carbohydrate-binding module families 1, 2 and 3. Then, we summarize four main kinds of methodologies for employing CBDs to modify cellulosic materials (i.e., CBD only, genetic fusion, non-covalent linkage and covalent linkage). Via different approaches, CBDs have been used to improve the material properties of cellulose, immobilize enzymes for biocatalysis, and design various detection tools. To achieve industrial applications, researches for lowering the production cost of CBDs, improving their performance (e.g., stability), and expanding their application scenarios are still in need. Graphical abstract

The continual emergence of highly pathogenic novel coronaviruses and their variants has underscored the importance of neutralizing monoclonal antibodies (mAbs) as a pivotal therapeutic approach. In the present study, we report the specific neutralizing antibodies 13H7 and 9G11, which target the N-terminal domain (NTD) and receptor-binding domain (RBD) of the SARS-CoV-2, respectively. The comparative analysis observed that 13H7 not only neutralizes early variants of concern (VOCs) but also exhibits neutralizing activity against the Omicron sublineage, including BA.4, BA.5, BQ.1, and BQ.1.1. 9G11, as an RBD antibody, also demonstrated remarkable neutralizing efficacy. A cocktail antibody combining 13H7 and 9G11 with the previously reported 3E2 broaden the neutralization spectrum against new variants of the SARS-CoV-2. We elucidated the cryo-EM structure of the complex, clarifying the mechanism of action of the cocktail antibody combination. Additionally, we also emphasized the molecular mechanism between 13H7 and SARS-CoV-2 NTD, revealing the impact of Y144 and H146 residue deletions and mutations on the neutralizing efficacy of 13H7. Taken together, our findings not only offer novel insights into the combination therapy of NTD and RBD neutralizing mAbs but also lay a theoretical foundation for the development of vaccines targeting NTD antibodies, thus providing valuable understanding of alleviating the emergence of SARS-CoV-2 variants.

Abstract Background This study aims to decipher the genetic basis governing yield components and quality attributes of peanuts, a critical aspect for advancing molecular breeding techniques. Integrating genotype re-sequencing and phenotypic evaluations of seven yield components and two grain quality traits across four distinct environments allowed for the execution of a genome-wide association study (GWAS). Results The nine phenotypic traits were all continuous and followed a normal distribution. The broad heritability ranged from 88.09 to 98.08%, and the genotype-environment interaction effects were all significant. There was a highly significant negative correlation between protein content (PC) and oil content (OC). The 10× genome re-sequencing of 199 peanut accessions yielded a total of 631,988 high-quality single nucleotide polymorphisms (SNPs), with 374 significant SNP loci identified in association with the nine traits of interest. Notably, 66 of these pertinent SNPs were detected in multiple environments, and 48 of them were linked to multiple traits of interest. Five loci situated on chromosome 16 (Chr16) exhibited pleiotropic effects on yield traits, accounting for 17.64–32.61% of the observed phenotypic variation. Two loci on Chr08 were found to be strongly associated with protein and oil contents, accounting for 12.86% and 14.06% of their respective phenotypic variations, respectively. Linkage disequilibrium (LD) block analysis of these seven loci unraveled five nonsynonymous variants, leading to the identification of one yield-related candidate gene and two quality-related candidate genes. The correlation between phenotypic variation and SNP loci in these candidate genes was validated by Kompetitive allele-specific PCR (KASP) marker analysis. Conclusions Overall, molecular markers were developed for genetic loci associated with yield and quality traits through a GWAS investigation of 199 peanut accessions across four distinct environments. These molecular tools can aid in the development of desirable peanut germplasm with an equilibrium of yield and quality through marker-assisted breeding.

Effects of climate warming on nutrition quality and storage of alpine grasslands are still controversial, which is not conducive to the management and utilization of alpine grasslands. A long-term warming experiment (with open-top chambers used to elevate temperature) was conducted at three elevations (relatively low, mid-, and high elevations with 4313, 4513, and 4693 m) of Northern Tibet in 2010 to compare the differences in forage nutritional quality and storage response to warming among three elevations and to explore the relationships between forage nutritional quality and production. In 2019, community surveys, observations of forage biomass and nutrition quality, and soil physicochemical properties were carried out. Forage nutrition quality included crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), ether extract (EE), crude ash (Ash), and water-soluble carbohydrate (WSC) content. Warming did not affect community aboveground biomass (AGB) at the three elevations. Warming improved community nutrition quality by increasing community CP content by 25.80% and decreasing community NDF content by 15.51% at the low elevation. In contrast, warming reduced community nutrition quality by increasing community CP, ADF, and NDF contents by 13.45%, 23.68%, and 17.43%, respectively, and decreasing Ash content by 39.50% at the high elevation. Warming did not affect community CP, ADF, NDF, EE, Ash, or WSC contents at the mid-elevation. Warming increased community nutrition storage by increasing community CP, ADF, and NDF storges by 74.69%, 88.18%, and 79.71%, respectively, at the high elevation. Warming did not affect community nutrition storages at the low or mid-elevations. Overall, forbs had higher CP, EE, Ash, and WSC contents and lower ADF and NDF contents compared with graminoids. Community EE content increased with community AGB, but community CP, ADF, NDF, EE, Ash, and WSC contents were not related to community AGB. Therefore, from the low to high elevation, the effects of warming on forage nutrition quality gradually changed from improving to inhibiting. Warming altered rangeland quality by affecting forage nutrition quality rather than forage production. There were no trade-offs between forage nutrition quality and forage production.

The present paper investigates the dynamic behaviors of a bubble restricted by two parallel plates near an elliptical wall. The typical experimental phenomena of the bubble are recorded employing the high-speed photography and a theoretical Kelvin impulse model is established. The impacts of the spatial position and the curvature of the wall on the bubble collapse behaviors are quantitatively investigated through the theoretical model and verified against the experimental results. The Kelvin impulse intensity and the direction during the bubble collapse process are compared and discussed for different elliptical-shaped walls. The main conclusions include: (1) During the bubble collapse process, the phenomenon of the bubble uneven splitting is discovered. (2) At different spatial positions and wall curvatures, the bubble collapse jet angle, movement distance, and velocity are in good agreement with the theoretical Kelvin impulse predictions. (3) As the short-to-long axis ratio increases, the differences in the distributions of the Kelvin impulse intensity and the direction near the elliptical wall gradually become larger, and the range of the influence of the impulse intensity expands.

Summary: The emergence of SARS-CoV-2 variants raises concerns about the efficacy of existing COVID-19 vaccines and therapeutics. Previously, we identified a conserved cryptic class 5 epitope of SARS-CoV-2 receptor binding domain (RBD) by two cross-neutralizing antibodies 7D6 and 6D6. Intriguingly, this site remains resistant to substantial mutations occurred in ever-changing SARS-CoV-2 subvariants. As compared to class 3 antibody S309, 6D6 maintains broad and consistent neutralizing activities against SARS-CoV-2 variants. Furthermore, 6D6 effectively protected hamster from the virulent Beta strain. Sequence alignment of approximately 6 million documented SARS-CoV-2 isolates revealed that 6D6 epitope maintains an exceptionally high conservation rate (99.92%). Structural analysis demonstrated that all 33 mutations accumulated in XBB.1.5 since the original strain do not perturb the binding 6D6 to RBD, in line with the sequence analysis throughout the antigenicity evolution of SARS-CoV-2. These findings suggest the potential of this epitope serving as a critical determinant for vaccines and therapeutic design.

The production performance and rumen bacterial diversity were compared for different silage-based diets supplemented with common concentrate or bio-concentrate to develop an alternative of common concentrate for fatten cattle feeding. The daily gain of fattening cattle was increased by 0. 99 kg and 1.04 kg, respectively, when fed with single corn silage or mixed silage-based diet supplemented with bio-concentrate. There was no significant difference in water loss rate and cooked meat rate among groups (P>0.05), but the tenderness of beef in the bio-concentrate group was significantly higher than that in the common concentrate group (P

Aerospace vehicle launched by electromagnetic propulsion is a potential option for future reusable space transportation systems. Complex ground effects and stability issues are induced generally due to the introduction of electromagnetic levitation force. A dynamic model coupled with electromagnetic force and aerodynamic force was established for the two-dimensional wing (NACA0012). Numerical simulations were conducted on the flow characteristics, operational attitude, and aerodynamic characteristics of the wing during the Ma＝0~1.5 acceleration process. It indicates that ground effects can be divided into four stages. In the first stage, subsonic flows are presented on both the upper and lower wing surfaces, and there is basically no oscillation for the attitude and aerodynamic loads of the wing. In the second stage, transonic flow emerges on the upper wing surface, while the flow on the lower wing surface is dominated by a typical variable cross-section transonic flow accompanied by the transition from the choked flow mode to the unchoked one. In the third stage, the upper wing surface maintains transonic flows, while the lower wing surface undergoes choked flows which are fully expanded. In both the second and third stages, the wing attitude and aerodynamic loads oscillate significantly at low frequencies. In the fourth stage, the upper wing surface undergoes supersonic flows, while the lower wing surface maintains choked flows. The wing attitude and aerodynamic loads oscillate slightly at high frequencies. On this basis, the effects of suspension height, suspension stiffness and spacing between suspension magnets on the system stability were explored. It is found that increa-sing the suspension height is beneficial for improving system stability. Increasing the suspension stiffness or spacing between suspension magnets appropriately, while limiting the target speed of electromagnetic propulsion to be less than the critical Mach number of system oscillation divergence, is beneficial for significantly improving system stability.

Introduction The escalating global changes have fostered conditions for the expansion and transmission of diverse biological factors, leading to the rise of emerging and reemerging infectious diseases. Complex viral infections, such as COVID-19, influenza, HIV, and Ebola, continue to surface, necessitating the development of effective vaccine technologies. Areas covered This review article highlights recent advancements in molecular biology, virology, and genomics that have propelled the design and development of innovative molecular tools. These tools have promoted new vaccine research platforms and directly improved vaccine efficacy. The review summarizes the cutting-edge molecular engineering tools used in creating novel vaccines and explores the rapidly expanding molecular tools landscape and potential directions for future vaccine development. Expert opinion The strategic application of advanced molecular engineering tools can address conventional vaccine limitations, enhance the overall efficacy of vaccine products, promote diversification in vaccine platforms, and form the foundation for future vaccine development. Prioritizing safety considerations of these novel molecular tools during vaccine development is crucial.

Introduction Vaccines are powerful tools for controlling microbial infections and preventing epidemics. To enhance the immune response to antigens, effective subunit vaccines or mRNA vaccines often require the combination of adjuvants or delivery carriers. In recent years, with the rapid development of immune mechanism research and nanotechnology, various studies based on the optimization of traditional adjuvants or various novel carriers have been intensified, and the construction of vaccine adjuvant delivery systems (VADS) with both adjuvant activity and antigen delivery has become more and more important in vaccine research. Areas covered This paper reviews the common types of vaccine adjuvant delivery carriers, classifies the VADS according to their basic carrier types, introduces the current research status and future development trend, and emphasizes the important role of VADS in novel vaccine research. Expert opinion As the number of vaccine types increases, conventional aluminum adjuvants show limitations in effectively stimulating cellular immune responses, limiting their use in therapeutic vaccines for intracellular infections or tumors. In contrast, the use of conventional adjuvants as VADS to carry immunostimulatory molecules or deliver antigens can greatly enhance the immune boosting effect of classical adjuvants. A comprehensive understanding of the various delivery vehicles will further facilitate the development of vaccine adjuvant research.

This study was based on QuEChERS cleanup coupled with UHPLC-MS/MS for the determination of γ-oryzanol compounds in vegetable oils. Several parameters of QuEChERS and UHPLC-MS/MS were studied for purification and detection of γ-oryzanol compounds in oil samples. Under the optimized conditions, the whole pretreatment procedure could be accomplished within 10 min without tedious procedure, larger volume of organic solvent and complicated apparatus. The limit of detections and the limit of quantifications for γ-oryzanol compounds were ranging from 0.1-0.3 µg kg−1 and 0.4–1.0 µg kg−1, respectively. Satisfactory recoveries of all analyts were ranging from 72.2 % to 101.3 %, and the intra-day and inter-day precision were less than 10.6 %. The validation indicated that rice band oil and corn oil were rich in 24-mCAF, CAF, β-SIF, CMF and STF. The QuEChERS-UHPLC-MS/MS simultaneously quantified five γ-oryzanol compounds in lipid matrices and assessed the nutritional and functional substances of vegetable oils.

Introduction: Protein engineering, which aims to improve the properties and functions of proteins, holds great research significance and application value. However, current models that predict the effects of amino acid substitutions often perform poorly when evaluated for precision. Recent research has shown that ProteinMPNN, a large-scale pre-training sequence design model based on protein structure, performs exceptionally well. It is capable of designing mutants with structures similar to the original protein. When applied to the field of protein engineering, the diverse designs for mutation positions generated by this model can be viewed as a more precise mutation range.Methods: We collected three biological experimental datasets and compared the design results of ProteinMPNN for wild-type proteins with the experimental datasets to verify the ability of ProteinMPNN in improving protein fitness.Results: The validation on biological experimental datasets shows that ProteinMPNN has the ability to design mutation types with higher fitness in single and multi-point mutations. We have verified the high accuracy of ProteinMPNN in protein engineering tasks from both positive and negative perspectives.Discussion: Our research indicates that using large-scale pre trained models to design protein mutants provides a new approach for protein engineering, providing strong support for guiding biological experiments and applications in biotechnology.

Abstract Tailed bacteriophages (order, Caudovirales) account for the majority of all phages. However, the long flexible tail of siphophages hinders comprehensive investigation of the mechanism of viral gene delivery. Here, we report the atomic capsid and in-situ structures of the tail machine of the marine siphophage, vB_DshS-R4C (R4C), which infects Roseobacter. The R4C virion, comprising 12 distinct structural protein components, has a unique five-fold vertex of the icosahedral capsid that allows genome delivery. The specific position and interaction pattern of the tail tube proteins determine the atypical long rigid tail of R4C, and further provide negative charge distribution within the tail tube. A ratchet mechanism assists in DNA transmission, which is initiated by an absorption device that structurally resembles the phage-like particle, RcGTA. Overall, these results provide in-depth knowledge into the intact structure and underlining DNA delivery mechanism for the ecologically important siphophages.

Abstract Background Rhizosphere soil physicochemical, endophytic fungi have an important role in plant growth. A large number of endophytic fungi play an indispensable role in promoting plant growth and development, and they can provide protection for host plants by producing a variety of secondary metabolites to resist and inhibit plant pathogens. Due to the terrain of Gansu province is north–south and longitudinal, different climatic conditions, altitude, terrain and growth environment will affect the growth of Codonopsis pilosula, and the changes in these environmental factors directly affect the quality and yield of C. pilosula in different production areas. However, In C. pilosula, the connection between soil nutrients, spatiotemporal variation and the community structure of endophytic fungi isolated from C. pilosula roots has not been well studied. Results Seven hundred six strains of endophytic fungi were obtained using tissue isolation and the hyphaend-purification method from C. pilosula roots that picked at all seasons and six districts (Huichuan, HC; Longxi, LX; Zhangxian, ZX; Minxian, MX; Weiyuan, WY; and Lintao, LT) in Gansu Province, China. Fusarium sp. (205 strains, 29.04%), Aspergillus sp. (196 strains, 27.76%), Alternaria sp. (73 strains, 10.34%), Penicillium sp. (58 strains, 8.22%) and Plectosphaerella sp. (56 strains, 7.93%) were the dominant genus. The species composition differed from temporal and spatial distribution (Autumn and Winter were higher than Spring and Summer, MX and LT had the highest similarity, HC and LT had the lowest). physical and chemical of soil like Electroconductibility (EC), Total nitrogen (TN), Catalase (CAT), Urease (URE) and Sucrase (SUC) had significant effects on agronomic traits of C. pilosula (P

Abstract Purpose To evaluate the effects of a novel technique using an isolated lens anterior capsule disc (LACD) to protect corneal endothelial cells in rabbit eyes during femtosecond laser-assisted cataract surgery. Methods Experimental study. 40 rabbits were divided into endothelium-protected (experimental) and control groups, with 20 rabbits in each group. In the experimental group, after femtosecond laser capsulotomy, the isolated capsule disc was lifted to the corneal endothelium by an ophthalmic viscosurgical device. The endothelium was damaged for 1 min with an ultrasonic probe. The control group underwent the same surgery, except that the disc was removed immediately after capsulorhexis. Corneal endothelioscopy was performed preoperatively and on postoperative days (PODs) 3 and 7 to observe endothelial cell counts (ECC) and endothelial cell loss rate. Central corneal thickness (CCT) was measured before and at PODs 1, 3 and 7. Results There were 3.59%±1.88% (p

Highlights An ion–electron coupled thermoelectric material was successfully prepared, which theoretically proved the ion–electron thermoelectric synergy effect and this material can work for a long time, which promoted low-grade thermal energy conversion. In the new operating mode of ion–electron thermoelectric synergy effect, our ionic thermoelectrics have a high Seebeck coefficient of 32.7 mV K−1 and a high energy density of 553.9 J m−2, enabling self-power for electronic components.

Currently, SARS-CoV-2 has evolved into various variants, including the numerous highly mutated Omicron sub-lineages, significantly increasing immune evasion ability. The development raises concerns about the possibly diminished effectiveness of available vaccines and antibody-based therapeutics. Here, we describe those representative categories of broadly neutralizing antibodies (bnAbs) that retain prominent effectiveness against emerging variants including Omicron sub-lineages. The molecular characteristics, epitope conservation, and resistance mechanisms of these antibodies are further detailed, aiming to offer suggestion or direction for the development of therapeutic antibodies, and facilitate the design of vaccines with broad-spectrum potential.

Infection caused by respiratory viruses can lead to a severe respiratory disease and even death. Vaccination is the most effective way to prevent the disease, but it cannot be quickly applied when facing an emerging infectious disease. Here, we demonstrated that immunization with an aluminium-zinc hybrid particulate adjuvant (FH-001) alone, bearing great resemblance in morphology with commonly used aluminium-based adjuvants in vaccines, could quickly induce mice to generate a broadly protective immune response to resist the lethal challenge of influenza B viruses. Furthermore, a multi-omics-based analysis revealed that the alveolar macrophage and type I interferon pathway, rather than adaptive immunity and type II interferon pathway, were essential for the observed prophylactic effect of FH-001. More importantly, a similar protective effect was observed against influenza A virus strain A/Shanghai/02/2013(H7N9), A/California/04/2009(H1N1) and respiratory syncytial virus. Therefore, we introduced here a new and promising strategy that can be quickly applied during the outbreak of emerging respiratory viruses.

In the working process of quadrupole composite rail electromagnetic launcher, there will be many phenomena such as structural deformation, temperature change and electromagnetic interference. At the same time, it involves the multi field effects of electromagnetic field, temperature field and structural field, and its action mechanism is quite complex. In this paper, the finite element analysis software ANSYS is used to simulate the electromagnetic structure, electromagnetic temperature and electromagnetic temperature structure of the quadrupole composite rail electromagnetic launcher, and to explore its multi physical field characteristics under the action of different factors. The simulation results show that the simulation model can reflect the actual phenomena in the process of electromagnetic launch. The different temperature rise of each component of the launcher at the end of the launch will cause different degrees of deformation of the armature and rail. Therefore, the influence of thermal stress should be fully considered when design-ing the device structure and adding excitation.

Abstract In vaccinology, a potent immunogen has two prerequisite attributes—antigenicity and immunogenicity. We have rational designed a triple-type HPV vaccine against HPV58, −33 and −52 covered in Gardasil 9 based on the sequence homology and similar surface loop structure of L1 protein, which is related to cross-type antigenicity. Here, we design another triple-type vaccine against non-vaccine types HPV39, −68 and −70 by immunogenicity optimization considering type specific immunodominant epitopes located in separate region for different types. First, we optimized the expression of wild-type HPV39, −68 and −70 L1-only virus-like particles (VLPs) in E. coli through N-terminal truncation of HPV L1 proteins and non-fusion soluble expression. Second, based on genetic relationships and an L1 homologous loop-swapping rationale, we constructed several triple-type chimeric VLPs for HPV39, −68 and −70, and obtained the lead candidate named H39–68FG-70DE by the immunogenicity optimization using reactivity profile of a panel type-specific monoclonal antibodies. Through comprehensive characterization using various biochemical, VLP-based analyses and immune assays, we show that H39–68FG-70DE assumes similar particulate properties as that of its parental VLPs, along with comparable neutralization immunogenicity for all three HPV types. Overall, this study shows the promise and translatability of an HPV39/68/70 triple-type vaccine, and the possibility of expanding the type-coverage of current HPV vaccines. Our study further expanded the essential criteria on the rational design of a cross-type vaccine, i.e. separate sites with inter-type similar sequence and structure as well as type-specific immunodominant epitope to be clustered together.

Although soil pH is an important indicator of soil quality, the effects of climate change and anthropogenic activities on soil pH remain controversial. Here, we quantified the potential soil pH at three depths (0–10, 10–20, and 20–30 cm) using annual temperature (AT), annual precipitation (AP) and annual radiation (ARad), and the actual soil pH at three depths using AT, AP, ARad and maximum normalized difference vegetation index based on random forest models over the grassland regions on the Tibetan Plateau in 2000–2020. Overall, climate change caused soil alkalinization at 0–10 cm, and soil acidification at 10–20 and 20–30 cm. Under climate change conditions, 36.84%, 29.87% and 23.71% regions showed soil alkalinization, whereas 45.52%, 44.49% and 21.43% regions showed soil acidification at 0–10, 10–20 and 20–30 cm, respectively. Climate change caused soil acidification in alpine meadows, but soil alkalinization in alpine steppes. The impacts of precipitation change on soil pH were not always greater than those of climate warming, depending on soil depth, and radiation change had some exclusive impacts on soil pH. Overall, anthropogenic activities caused soil alkalinization, but more than 1/10 regions showed soil acidification. Anthropogenic activities caused soil acidification in alpine meadows, but soil alkalinization in montane meadows at 10–20 cm. Therefore, the impacts of climate change and anthropogenic activities on soil pH did not always cause soil acidification or alkalization, but varied with grassland type and soil depth. Both climate change and anthropogenic activities reconstructed the spatial distribution pattern of soil pH. This study also cautioned that the impact of radiation change on soil pH should not be ignored.

Circulating subtypes of Influenza viruses seasonally change and therefore vaccines need to be matched to these strains each year, which is why there is a need for next-generation vaccines that can elicit broad and cross-type protection. Here, Li et al. generate a human-mouse chimeric antibody with broad neutralizing activity against seasonal and pandemic H1N1 and some H5N1 viruses in vivo and identify residues on hemagglutinin relevant for its broad neutralization activity.

Abstract The major challenge to controlling the COVID pandemic is the rapid mutation rate of the SARS-CoV-2 virus, leading to the escape of the protection of vaccines and most of the neutralizing antibodies to date. Thus, it is essential to develop neutralizing antibodies with broad-spectrum activity targeting multiple SARS-CoV-2 variants. Here, we report a synthetic nanobody (named C5G2) obtained by phage display and subsequent antibody engineering. C5G2 has a single-digit nanomolar binding affinity to the RBD domain and inhibits its binding to ACE2 with an IC50 of 3.7 nM. Pseudovirus assays indicated that monovalent C5G2 could protect the cells from infection with SARS-CoV-2 wild-type virus and most of the viruses of concern, i.e., Alpha, Beta, Gamma and Omicron variants. Strikingly, C5G2 has the highest potency against Omicron BA.1 among all the variants, with an IC50 of 4.9 ng/mL. The cryo-EM structure of C5G2 in complex with the spike trimer showed that C5G2 binds to RBD mainly through its CDR3 at a conserved region that does not overlap with the ACE2 binding surface. Additionally, C5G2 binds simultaneously to the neighboring NTD domain of the spike trimer through the same CDR3 loop, which may further increase its potency against viral infection. Third, the steric hindrance caused by FR2 of C5G2 could inhibit the binding of ACE2 to RBD as well. Thus, this triple-function nanobody may serve as an effective drug for prophylaxis and therapy against Omicron as well as future variants.

IntroductionParental stress among primary caregivers of children with autism spectrum disorder (ASD) is a significant concern. While previous research indicates that both family and child factors substantially influence parental stress, a few studies have comprehensively examined these factors from family, parent, and child perspectives. Moreover, the psychological mechanisms underlying parental stress remain underexplored.MethodThis study obtained a valid sample of 478 primary caregivers of children diagnosed with ASD in China and employed mediation and moderated mediation analyses to investigate the relationships between family adaptability and cohesion (FAC), ASD severity, parental self-efficacy, and parental stress.ResultResults revealed that higher FAC was linked to reduced parental stress through increased parental self-efficacy. The indirect effect of parental self-efficacy was more substantial for caregivers of children with severe symptoms than those with mild symptoms.DiscussionThese findings offer insights into how FAC influences parental stress and underscore the importance of parental self-efficacy as a coping resource for mitigating parental stress. This study provides valuable theoretical and practical implications for understanding and addressing parental stress, particularly in families raising children with ASD.

Pseudorabies virus (PRV) is a major etiological agent of swine infectious diseases responsible for significant economic losses in the swine industry. The authors report the structures of the PRV A-capsid and C-capsid, shedding light on PRV’s assembly mechanism.

In the past 30 years, the small baseline subset (SBAS) InSAR time-series technique has emerged as an essential tool for measuring slow surface displacement and estimating geophysical parameters. Because of its ability to monitor large-scale deformation with millimeter accuracy, the SBAS method has been widely used in various geodetic fields, such as ground subsidence, landslides, and seismic activity. The obtained long-term time-series cumulative deformation is vital for studying the deformation mechanism. This article reviews the algorithms, applications, and challenges of the SBAS method. First, we recall the fundamental principle and analyze the shortcomings of the traditional SBAS algorithm, which provides a basic framework for the following improved time series methods. Second, we classify the current improved SBAS techniques from different perspectives: solving the ill-posed equation, increasing the density of high-coherence points, improving the accuracy of monitoring deformation and measuring the multi-dimensional deformation. Third, we summarize the application of the SBAS method in monitoring ground subsidence, permafrost degradation, glacier movement, volcanic activity, landslides, and seismic activity. Finally, we discuss the difficulties faced by the SBAS method and explore its future development direction.

Conformational changes or rearrangements are common events during inter-biomolecular recognition. Tracking these changes are essential for exploring the allosteric mechanism and it is usually achieved by molecular dynamics simulation in silico. We previously identified a broad-neutralizing antibody against H5 influenza virus, 13D4, and solved the crystal structures of the free 13D4 Fab and its complex with hemagglutinin (HA). Structural comparison of the unbound and bound 13D4 Fabs showed that the heavy chain complementarity-determining region 3 (HCDR3) undergoes a substantial conformational rearrangement when it recognizes the receptor-binding site (RBS). Here, we used molecular dynamics (MD) to simulate the conformational changes that occur during antibody recognition. We showed that neither conventional MD nor steered MD could recapitulate the loop fitting of the RBS structure contour. Consequently, to simulate these challenging conformational changes, we engaged a stepwise docking MD method that allowed for the gradual docking of the ligand to receptor. This new method recapitulates the bound shape of the HCDR3 and provides the best approximation of the shape rendered by the co-crystal structure, with an RMSD of 0.926 Å. This strategy affords a flexible MD approach for simulating complicated conformational changes that occur during molecular recognition, and helps to provide an understanding of the involved allosteric mechanism.

IntroductionAs one of the most common malignant tumors in clinical practice, hepatocellular carcinoma (HCC) is a major threat to human health, where alpha-fetoprotein (AFP) is widely used for early screening and diagnoses. However, the level of AFP would not elevate in about 30-40% of HCC patients, which is clinically referred to as AFP-negative HCC, with small tumors at an early stage and atypical imaging features, making it difficult to distinguish benign from malignant by imaging alone.MethodsA total of 798 patients, with the majority being HBV-positive, were enrolled in the study and were randomized 2:1 to the training and validation groups. Univariate and multivariate binary logistic regression analyses were used to determine the ability of each parameter to predict HCC. A nomogram model was constructed based on the independent predictors. ResultsA unordered multicategorical logistic regression analyses showed that the age, TBIL, ALT, ALB, PT, GGT and GPR help identify non-hepatic disease, hepatitis, cirrhosis, and hepatocellular carcinoma. A multivariate logistic regression analyses showed that the gender, age, TBIL, GAR, and GPR were independent predictors for the diagnosis of AFP-negative HCC. And an efficient and reliable nomogram model (AUC=0.837) was constructed based on independent predictors.DiscussionSerum parameters help reveal intrinsic differences between non-hepatic disease, hepatitis, cirrhosis, and HCC. The nomogram based on clinical and serum parameters could be used as a marker for the diagnosis of AFP-negative HCC, providing an objective basis for the early diagnosis and individualized treatment of hepatocellular carcinoma patients.

Tribotronics represents the modulation of friction via an external electric potential, a field with promising ramifications for intelligent devices, precision manufacturing, and biomedical applications. A profound elucidation of mechanisms that allow for potential-controlled friction is foundational to further research in this tribotronic domain. This article provides a comprehensive review of the research progress in electro-controlled friction over the past few decades, approached from the perspective of the boundary lubrication film at the friction interface, a direct influencer of electro-controlled friction performance. The mechanisms of potential-controlled friction are categorized into three distinct classifications, contingent on the formation mode of the boundary lubrication film: potential-induced interfacial redox reactions, interfacial physical adsorption, and interfacial phase structure transformations. Furthermore, an outlook on the application prospects of electro-controlled friction is provided. Finally, several research directions worth exploring in the field of electro-controlled friction are proposed. The authors hope that this article will further promote the application of electro-controlled friction technology in engineering and provide intellectual inspiration for related researchers.

Abstract Background The various advantages associated with the growth properties of Escherichia coli have justified their use in the production of genetically engineered vaccines. However, endotoxin contamination, plasmid vector instability, and the requirement for antibiotic supplementation are frequent bottlenecks in the successful production of recombinant proteins that are safe for industrial-scaled applications. To overcome these drawbacks, we focused on interrupting the expression of several key genes involved in the synthesis of lipopolysaccharide (LPS), an endotoxin frequently responsible for toxicity in recombinant proteins, to eliminate endotoxin contamination and produce better recombinant proteins with E. coli. Results Of 8 potential target genes associated with LPS synthesis, we successfully constructed 7 LPS biosynthesis-defective recombinant strains to reduce the production of LPS. The endotoxin residue in the protein products from these modified E. coli strains were about two orders of magnitude lower than that produced by the wild-type strain. Further, we found that 6 loci—lpxM, lpxP, lpxL, eptA, gutQ and kdsD—were suitable for chromosomal integrated expression of HPV L1 protein. We found that a single copy of the expression cassette conferred stable expression during long-term antibiotic-free cultivation as compared with the more variable protein production from plasmid-based expression. In large-scale fermentation, we found that recombinant strains bearing 3 to 5 copies of the expression cassette had 1.5- to 2-fold higher overall expression along with lower endotoxin levels as compared with the parental ER2566 strain. Finally, we engineered and constructed 9 recombinant E. coli strains for the later production of an HPV 9-valent capsid protein with desirable purity, VLP morphology, and antigenicity. Conclusions Reengineering the LPS synthesis loci in the E. coli ER2566 strain through chromosomal integration of expression cassettes has potential uses for the production of a 9-valent HPV vaccine candidate, with markedly reduced residual endotoxin levels. Our results offer a new strategy for recombinant E. coli strain construction, engineering, and the development of suitable recombinant protein drugs.

Thimerosal has been widely used as a preservative in drug and vaccine products for decades. Due to the strong propensity to modify thiols in proteins, conformational changes could occur due to covalent bond formation between ethylmercury (a degradant of thimerosal) and thiols. Such a conformational change could lead to partial or even complete loss of desirable protein function. This study aims to investigate the effects of thimerosal on the capsid stability and antigenicity of recombinant human papillomavirus (HPV) 18 virus-like particles (VLPs). Dramatic destabilization of the recombinant viral capsid upon thimerosal treatment was observed. Such a negative effect on the thermal stability of VLPs preserved with thimerosal was shown to be dependent on the thimerosal concentration. Two highly neutralizing antibodies, 13H12 and 3C3, were found to be the most sensitive to thimerosal treatment. The kinetics of antigenicity loss, when monitored with 13H12 or 3C3 as probes, yielded two distinctly different sets of kinetic parameters, while the data from both monoclonal antibodies (mAbs) followed a biphasic exponential decay model. The potential effect of thimerosal on protein function, particularly for thiol-containing proteinaceous active components, needs to be comprehensively characterized during formulation development when a preservative is necessary.

The Qinghai-Tibet Plateau has experienced an increase in N deposition/input due to global change. However, it remains unclear how the responses and whether the responses of soil bacterial diversity to short-term N input are consistent at different levels. Here, we investigated soil bacterial species and phylogenetic α-diversity and community composition based on a short-term nitrogen input experiment (five levels: 0, 2.5, 5, 10, and 20 g N m−2 y−1) in an alpine steppe at the source of Brahmaputra, using high-throughput sequencing technology. Short-term nitrogen input did not affect the species α-diversity and β-diversity of soil bacteria. However, soil bacterial phylogenetic α-diversity and dissimilarity increased with increasing nitrogen input. Different relative contributions and correlations of primary factors to species and phylogenetic diversity under short-term nitrogen input may result in different responses, in which ecological processes also play a role. Therefore, studying the response of soil bacteria to short-term nitrogen input should take into account not only the species level but also the phylogenetic level. We should pay close attention to the potential influence of short-term nitrogen deposition/fertilization on the soil bacterial community in the alpine steppe on the Tibetan Plateau.

Objective: The incidence of post-traumatic stress disorder (PTSD) increased among healthcare workers (HCWs) during the outbreak of COVID-19. The purpose of this study was to examine the relationship between eating behavior and PTSD, considering the mediation effect of anxiety, depression and sleep. Methods: A total of 101 HCWs completed a survey. The Food-Frequency Questionnaires (FFQ) were used to evaluate the diet. A special survey was conducted on the eating time of each shift mode. The PTSD Checklist-Civilian Version (PCL-C), Self-Rating Anxiety Scale (SAS), Self-Rating Depression Scale (SDS), Pittsburgh Sleep Quality Index (PSQI), and Morning-Evening Questionnaire (MEQ) were utilized to assess clinical symptoms. Results: There was a statistically significant correlation between the night shift eat midpoint (NEMP) and PTSD symptoms, anxiety and depression as significant mediators. The last meal jet lag between night shift and day shift (NDLM) was related to PTSD symptoms significantly, and sleep and anxiety were significant mediators. The relationship between animal-based protein pattern and PTSD symptoms was statistically significant, and anxiety was the significant mediator. Conclusions: The earlier the HCWs eat in the night shift, the lighter the symptoms of PTSD. This is mediated by improving anxiety, depression and sleep disorder. Furthermore, the consumption of animal protein could reduce symptoms of PTSD by improving anxiety.

Antibodies (Abs) targeting highly conserved epitopes are important tools against emerging virus variants. Here, the authors characterize Abs that recognize a cryptic epitope in the receptor-binding domain of SARS-CoV-2 spike that is well conserved and show that these Abs can neutralize several variants of concerns.

Surface plasmons have unique physical properties that make them also interesting for technology. Here, the authors observe plasmons in mixed-dimensional heterostructures that can be highly modulated with electrostatic gating, which may be explained by plasmon hybridization