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Abstract Nickel-rich layered oxides are one of the most promising positive electrode active materials for high-energy Li-ion batteries. Unfortunately, the practical performance is inevitably circumscribed by the structural deterioration deriving from the Ni/Li antisite disorder, leading to severe capacity loss and life attenuation. Herein, we propose an economical and facile rejuvenation strategy by employing the magneto-electrochemical synergistic activation targeting the positive electrode in assembled Li-ion batteries. This approach induces a transition of Ni3+ from high-spin to low-spin, reducing the super-exchange interaction of Ni-O-transition metal (TM). Meanwhile, electrochemical reaction drives Li+ from the host material and promotes Ni3+ to reoccupy TM layer, recovering intrinsic Li site and extending cycle life. The strategy demonstrates that low-quality positive electrodes can be converted to high-quality ones. Notably, the method can revitalize an aged Li-ion pouch cell (SiC||NCM811, 8 Ah nominal capacity) via optimizing cation occupancy and increase its capacity by 10% from 6.49 to 7.14 Ah at 1 C, illustrating the benefits of the upcycling process.

Highlights A multiscale hierarchical structure design, integrating wrinkled MXene radar-infrared shielding layer and flexible Fe3O4@C/PDMS microwave absorption layer The assembled stealth devices realize a near-perfect stealth capability in both X-band (8-12 GHz) and long-wave infrared (8-14 µm) wavelength ranges. The optimal device demonstrates exceptional curved surface conformability, self-cleaning capability (contact angle ≈ 129°), and abrasion resistance (recovery time ≈ 5 s).

Black phosphorus (BP) anode with high capacity (2596 mAh g−1) and suitable lithiation potential (0.7 ​V vs. Li+/Li) is an ideal candidate for high-energy-density and high-safety lithium-ion batteries, however, the practical implementation is greatly limited by its slow reaction kinetics and huge volume expansion. Here, inspired by nature, liquid metal (LM) is explored as a self-heal agent, which can well stabilize the BP anode through buffering the volumetric expansion and re-activating “dead P and LixP”. Moreover, LM also acts as a good catalyst, which can adjust Li ion concentration and reduce the activation energy of delithiation reaction, thus prolonging the cycling life. Therefore, the LM modified BP/graphite (G) composite delivers an excellent high-rate performance of 1123 mAh g−1 ​at 4 ​C with 80.0 ​% capacity retention after 200 cycles, a superior wide-temperature performance of 1547.5 mAh g−1 and 569.0 mAh g−1 ​at 50 ​°C and −20 ​°C, respectively.

Summary: Broadly neutralizing antibodies (bNAbs) targeting the apex of the HIV-1-envelope (Env) trimer comprise the most potent category of HIV-1 bNAbs and have emerged as promising therapeutics. Here, we investigate the development of the HIV-1 apex-directed PGT145-PGDM1400 antibody lineage and report cryo-EM structures at 3.4 Å resolution of PGDM1400 and of an improved PGT145 variant (PGT145-R100aS), each bound to the BG505 Env trimer. Cross-species-based engineering improves PGT145 IC80 breadth to near that of PGDM1400. Despite similar breadth and potency, the two antibodies differ in their residue-level interactions with important apex features, including N160 glycans and apex cavity, with residue 100i of PGT145 (sulfated tyrosine) penetrating ∼7 Å farther than residue 100i of PGDM1400 (aspartic acid). While apex-directed bNAbs from other donors use maturation pathways that often converge on analogous residue-level recognition, our results demonstrate that divergent residue-level recognition can occur within the same lineage, thereby enabling improved coverage of escape variants.

Maintaining grassland stock and improving its ecosystem services (ES) are critical yet challenging for ecological and economic sustainability. However, the diversity and spatial heterogeneity of global grasslands pose significant challenges to studying the spatio-temporal coupling characteristics of grassland area and ES changes, especially in the karst desertification control (KDC) regions. This study quantified the grassland area, grassland ES, and the elasticity of grassland ES in response to changes in grassland area within the KDC region of South China. The Moran index was applied to analyze spatio-temporal evolution characteristics, and a quantile regression model was used to explored the segmentation effects of influencing factors. Results showed that: (1) During 2000–2020, despite a 4.36 % decrease in grassland area (399,205 hm2) within the KDC region of South China, the comprehensive index of grassland ES increased from 0.99 to 1.03. Furthermore, counties where the elasticity index of grassland ES exceeded 1 constituted 73.52 %, indicating a general improvement in ES supply capacity. (2) Factors influencing grassland ES in the KDC region demonstrated clear segmentation effects, with the intensity and direction of the same factors varying significantly across different thresholds. The significance of socio-economic factors has been increasing steadily. (3) Within the framework of the UN Decade on Ecosystem Restoration 2021–2030, differentiated ecological engineering measures should be tailored based on the specific constraints of grassland ES in the KDC region. Our research results open a new perspective for the study of the impact mechanism of grassland ES. It can enrich the understanding of grassland ES and offer valuable insights to guide future initiatives for ecosystem restoration and sustainable development in ecologically fragile, particularly karst, areas.

In this study, headspace solid phase micro-extraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS) was adopted to separate and identify the volatile components of Kombucha by using polar column DB-WAX. Then, odor activity value (OAV) combined with stoichiometry was used to explore the volatile substance composition and characteristic differential substances of Kombucha in different regions. The results showed that there were significant differences between the volatile substances of Kombucha in different regions. A total of 56 kinds of volatile substances were detected, mainly alcohols, acids, esters and phenols. The OAV results showed that 32 kinds of substances contributed to the flavor for Kombucha (OAV≥1), the OAV of ethyl isovalerate of Kombucha in different regions was highest except for Urumqi and Weihai, it might play an important role in the flavor composition of Kombucha in different regions. Furthermore, with partial least squares discrimination analysis (PLS-DA), 17 variables with importance values greater than 1 as differential markers were identifed. By integrating the results of OAV and PLS-DA, a total of 14 characteristic compounds of Kombucha were screened out, namely 3-methyl-1-butanol, 2-ethyl-1-hexanol, phenethyl alcohol, nonanol, acetic acid, 3-methylbutyric acid, caprylic acid, ethyl acetate, isopentyl acetate, ethyl caprylate, ethyl phenylacetate, furan formaldehyde, 2,6-di-tert-butyl-p-cresol, and 4-ethylphenol. The research results will provide theoretical reference for the standardization, high-quality production and construction of characteristic brands of Kombucha.

Summary: Antibodies that target the gp120-gp41 interface of the HIV-1 envelope (Env) trimer comprise a commonly elicited category of broadly neutralizing antibodies (bNAbs). Here, we isolate and characterize VRC44, a bNAb lineage with up to 52% neutralization breadth. The cryoelectron microscopy (cryo-EM) structure of antibody VRC44.01 in complex with the Env trimer reveals binding to the same gp120-gp41 interface site of vulnerability as antibody 35O22 from a different HIV-1-infected donor. In addition to having similar angles of approach and extensive contacts with glycans N88 and N625, VRC44 and 35O22 derive from the same IGHV1-18 gene and share convergent mutations, indicating these two antibodies to be members of the only known highly glycan-dependent multidonor class. Strikingly, both lineages achieved almost 100% neutralization breadth against virus strains displaying high-mannose glycans. The high breadth and reproducible elicitation of VRC44 and 35O22 lineages validate germline-based methods of immunogen design for targeting the HIV-1 gp120-gp41 interface.

Broadly neutralizing antibodies have been proposed as templates for HIV-1 vaccine design, but it has been unclear how similar vaccine-elicited antibodies are to their naturally elicited templates. To provide insight, here we compare the recognition of naturally elicited and vaccine-elicited antibodies targeting the HIV-1 fusion peptide, which comprises envelope (Env) residues 512–526, with the most common sequence being AVGIGAVFLGFLGAA. Naturally elicited antibodies bound peptides with substitutions to negatively charged amino acids at residue positions 517–520 substantially better than the most common sequence, despite these substitutions rarely appearing in HIV-1; by contrast, vaccine-elicited antibodies were less tolerant of sequence variation, with no substitution of residues 512–516 showing increased binding. Molecular dynamics analysis and cryo-EM structural analysis of the naturally elicited ACS202 antibody in complex with the HIV-1 Env trimer with an alanine 517 to glutamine substitution suggested enhanced binding to result from electrostatic interactions with positively charged antibody residues. Overall, vaccine-elicited antibodies appeared to be more fully optimized to bind the most common fusion peptide sequence, perhaps reflecting the immunization with fusion peptide of the vaccine-elicited antibodies.

Summary: Background: Cyclin-dependent kinase 4/6 (CDK4/6) inhibitors in combination with traditional endocrine therapy (ET) are now the recommended first-line treatment for hormone receptor (HR)-positive and HER2-negative metastatic breast cancer (MBC). However, the benefits of adding CDK4/6 inhibitors to ET in HER2-low-positive and HER2-0 subgroups remain unclear. We aimed to assess the effectiveness of CDK4/6 inhibitors in combination with ET in patients with HR-positive, HER2-low-positive and HER2-0 MBC. Methods: This secondary analysis assessed progression-free survival (PFS) among HER2-low-positive and HER2-0 patients enrolled in the double-blind, placebo-controlled randomised clinical trials PALOMA-2 and PALOMA-3. The study included 1186 HER2-negative, HR-positive female patients, with available immunohistochemistry (IHC) and/or in situ hybridization (ISH) results, across 17 countries enrolled between February 2013 and August 2014. HER2-low-positive status was defined by IHC 1+ or 2+ with negative ISH, and HER2-zero by IHC 0. Data analyses were conducted between March and May 2023. In the PALOMA-2 trial, patients were randomly assigned to receive either palbociclib or placebo, in combination with letrozole in the first-line treatment for HR-positive MBC. Patients in the PALOMA-3 study, who had progression or relapse during previous ET, were randomly allocated to receive either palbociclib plus fulvestrant or placebo plus fulvestrant. The primary endpoint was investigator-assessed PFS. Kaplan–Meier approach and Cox proportional hazards model were applied to estimate the association of treatment strategies with PFS among HER2-0 and HER2-low-positive populations. The two trials are registered with ClinicalTrials.gov, number NCT01740427 and NCT01942135. Findings: Of the 666 patients with MBC from the PALOMA-2 study, there were 153 HER2-0 and 513 HER2-low-positive patients. In the HER2-0 population, no significant difference in PFS was observed between the palbociclib-letrozole and placebo-letrozole groups (hazard ratio = 0.79, 95% confidence interval [CI] 0.48–1.30, p = 0.34). In the HER2-low-positive population, palbociclib-letrozole demonstrated a significantly lower risk of PFS than placebo-letrozole group (hazard ratio = 0.52, 95% CI 0.41–0.66, p

Abstract Broadly neutralizing antibodies are proposed as therapeutic and prophylactic agents against HIV‐1, but their potency and breadth are less than optimal. This study describes the immunization of a llama with the prefusion‐stabilized HIV‐1 envelope (Env) trimer, BG505 DS‐SOSIP, and the identification and improvement of potent neutralizing nanobodies recognizing the CD4‐binding site (CD4bs) of vulnerability. Two of the vaccine‐elicited CD4bs‐targeting nanobodies, G36 and R27, when engineered into a triple tandem format with llama IgG2a‐hinge region and human IgG1‐constant region (G36×3‐IgG2a and R27×3‐IgG2a), neutralized 96% of a multiclade 208‐strain panel at geometric mean IC80s of 0.314 and 0.033 µg mL−1, respectively. Cryo‐EM structures of these nanobodies in complex with Env trimer revealed the two nanobodies to neutralize HIV‐1 by mimicking the recognition of the CD4 receptor. To enhance their neutralizing potency and breadth, nanobodies are linked to the light chain of the V2‐apex‐targeting broadly neutralizing antibody, CAP256V2LS. The resultant human‐llama bispecific antibody CAP256L‐R27×3LS exhibited ultrapotent neutralization and breadth exceeding other published HIV‐1 broadly neutralizing antibodies, with pharmacokinetics determined in FcRn‐Fc mice similar to the parent CAP256V2LS. Vaccine‐elicited llama nanobodies, when combined with V2‐apex broadly neutralizing antibodies, may therefore be able to fulfill anti‐HIV‐1 therapeutic and prophylactic clinical goals.

Abstract Lassa virus (LASV) infection is expanding outside its traditionally endemic areas in West Africa, posing a pandemic biothreat. LASV-neutralizing antibodies, moreover, have proven difficult to elicit. To gain insight into LASV neutralization, here we develop a prefusion-stabilized LASV glycoprotein trimer (GPC), pan it against phage libraries comprising single-domain antibodies (nanobodies) from shark and camel, and identify one, D5, which neutralizes LASV. Cryo-EM analyses reveal D5 to recognize a cleavage-dependent site-of-vulnerability at the trimer apex. The recognized site appears specific to GPC intermediates, with protomers lacking full cleavage between GP1 and GP2 subunits. Guinea pig immunizations with the prefusion-stabilized cleavage-intermediate LASV GPC, first as trimer and then as a nanoparticle, induce neutralizing responses, targeting multiple epitopes including that of D5; we identify a neutralizing antibody (GP23) from the immunized guinea pigs. Collectively, our findings define a prefusion-stabilized GPC trimer, reveal an apex-situated site-of-vulnerability, and demonstrate elicitation of LASV-neutralizing responses by a cleavage-intermediate LASV trimer.

With the deepening application of artificial intelligence in the field of education, teaching quality evaluation has become the key to improving educational effectiveness. In order to quantify the effectiveness of knowledge transmission and mastery in digital media teaching, this paper proposes an improved Bayesian knowledge tracking model (BF-BKT), which has been specifically optimized for teaching quality evaluation. The motivation stems from the shortcomings of existing technology in evaluating teaching quality, especially the lack of effective integration of student learning behavior and forgetting patterns. This article describes the development process of the BF-BKT model, which innovatively combines learning behavior data and forgetting patterns by introducing behavioral forgetting features to more accurately predict students’ learning status and teaching effectiveness. The BF-BKT model not only considers whether students have mastered a certain knowledge point, but also further analyzes their forgetting situation at different time points, providing a more comprehensive perspective for evaluating teaching quality. To validate the performance of the BF-BKT model, we compared it with the traditional BKT model and two diverse BKT variants. We conducted systematic experiments and analysis by selecting public datasets as simulation scenarios. The experimental results show that the BF-BKT model performs well in predicting students’ knowledge mastery and evaluating teaching quality, and its performance is superior to other comparative methods, proving the effectiveness and superiority of the model.

Abstract The HIV-1 fusion peptide (FP) represents a promising vaccine target, but global FP sequence diversity among circulating strains has limited anti-FP antibodies to ~60% neutralization breadth. Here we evolve the FP-targeting antibody VRC34.01 in vitro to enhance FP-neutralization using site saturation mutagenesis and yeast display. Successive rounds of directed evolution by iterative selection of antibodies for binding to resistant HIV-1 strains establish a variant, VRC34.01_mm28, as a best-in-class antibody with 10-fold enhanced potency compared to the template antibody and ~80% breadth on a cross-clade 208-strain neutralization panel. Structural analyses demonstrate that the improved paratope expands the FP binding groove to accommodate diverse FP sequences of different lengths while also recognizing the HIV-1 Env backbone. These data reveal critical antibody features for enhanced neutralization breadth and potency against the FP site of vulnerability and accelerate clinical development of broad HIV-1 FP-targeting vaccines and therapeutics.

IntroductionPinus koraiensis is a dominant tree species in northeastern China. Estimating its biomass is required for forest carbon stock monitoring and accounting.MethodsThis study investigates biomass estimation methods for P. koraiensis components. A Bayesian approach was used to synthesize the parameter distributions of 298 biomass models as prior information to estimate the trunk, branch, leaf, and root biomass of P. koraiensis. The results were compared with non-informative prior and the minimum least squares (MLS).ResultsThe results indicated that the Bayesian approach outperformed the other methods regarding model fit and prediction error. In addition, the responses of different components to tree height varied. The models of trunk and root biomass exhibited a smaller response to tree height, whereas those of branches and leaves showed a larger response to tree height. The model parameters yield precise estimations.DiscussionIn sum, this study highlights the potential of the Bayesian methods in estimating P. koraiensis biomass and proposes further enhancements to improve estimation accuracy.

In this paper, konjac powde were hydrolyzed by Bacillus licheniformis to prepare oligosaccharides. The effects of concentration of konjac powder, inoculum amount of strain, bacteriolysis temperature and hydrolysis time on hydrolysis rate of konjac powder were determined by single factor tests. The antioxidant activity of konjac powder oligosaccharides was evaluated with the scavenging ability of ·OH, ·O2−· and DPPH· as indexes. The prebiotic activity of konjac powder oligosaccharides was investigated by in vitro fermentation test. The results showed that the optimal hydrolysis process parameters was as follows: concentration of konjac powder 20 g/L, inoculation capacity 1×107 CFU/g, hydrolysis temperature 43.6 ℃, hydrolysis time 10.2 h. Under the optimal conditions, the hydrolysis rate of konjac powder was 39.06%±0.12%, and the yield of KGOS was 66.72%. With the prolongation of hydrolysis time, the viscosity of solution decreased from 2430.33 mPa·s to 18.06 mPa·s. When the concentration of konjac powder was 10 mg/mL, the clearance rates of ·OH, ·O2− and DPPH· were 61.47%, 34.60% and 81.43%, respectively. The OD600 of KGOS, glucose and konjac powder on the proliferation of Lactobacillus plantarum, Lactobacillus brevity and Lactobacillus casei in vitro were 0.775, 1.371, 0.236, 0.791, 1.339, 0.224 and 0.916, 1.336, 0.243, respectively. The results provide a theoretical basis for the further processing and utilization of oligosaccharides in konjac powder.

Abstract While several COVID-19 vaccines have been in use, more effective and durable vaccines are needed to combat the ongoing COVID-19 pandemic. Here, we report highly immunogenic self-assembling SARS-CoV-2 spike-HBsAg nanoparticles displaying a six-proline-stabilized WA1 (wild type, WT) spike S6P on a HBsAg core. These S6P-HBsAgs bound diverse domain-specific SARS-CoV-2 monoclonal antibodies. In mice with and without a HBV pre-vaccination, DNA immunization with S6P-HBsAgs elicited significantly more potent and durable neutralizing antibody (nAb) responses against diverse SARS-CoV-2 strains than that of soluble S2P or S6P, or full-length S2P with its coding sequence matching mRNA-1273. The nAb responses elicited by S6P-HBsAgs persisted substantially longer than by soluble S2P or S6P and appeared to be enhanced by HBsAg pre-exposure. These data show that genetic delivery of SARS-CoV-2 S6P-HBsAg nanoparticles can elicit greater and more durable nAb responses than non-nanoparticle forms of stabilized spike. Our findings highlight the potential of S6P-HBsAgs as next generation genetic vaccine candidates against SARS-CoV-2.

Objectives: The design of cancer clinical trials incorporating biomarkers depends on various factors, including the trial phase, the type of biomarker and whether its role has been validated. This article aims to present a method for designing and analyzing phase II cancer clinical trials that validate predictive biomarkers. Methods: We propose a randomized trial design where patients are allocated between a targeted therapy and a non-targeted therapy stratified by biomarker status. Tumor response is used as the primary endpoint to validate the biomarker through interaction testing between treatment and biomarker positivity. Additionally we propose a sample size calculation method for this design, considering two types of interaction: one based on logit-transformed response rates and the other on raw response rates. Results: The proposed sample size method is applied to the design of a real randomized phase II trial. Extensive simulations are conducted to evaluate the performance of the test statistic and the sample size method under different scenarios. Conclusions: Our method provides a practical approach to validating predictive biomarkers in phase II cancer trials. The simulations demonstrate robust performance for both interaction models, offering guidance for the sample size selection and analysis strategy in biomarker-stratified trials.

Abstract Development of a malaria vaccine that blocks transmission of different parasite stages to humans and mosquitoes is considered critical for elimination efforts. A vaccine using Pfs25, a protein on the surface of zygotes and ookinetes, is under investigation as a transmission-blocking vaccine (TBV) that would interrupt parasite passage from mosquitoes to humans. The most extensively studied Pfs25 TBVs use Pichia pastoris-produced recombinant forms of Pfs25, chemically conjugated to a recombinant carrier protein, ExoProtein A (EPA). The recombinant form of Pfs25 first used in humans was identified as Pfs25H, which contained a total of 14 heterologous amino acid residues located at the amino- and carboxyl-termini including a His6 affinity tag. A second recombinant Pfs25, identified as Pfs25M, was produced to remove the heterologous amino acid residues and conjugated to EPA (Pfs25M-EPA). Here, monomeric Pfs25M was characterized biochemically and biophysically for identity, purity, and integrity including protein structure to assess its comparability with Pfs25H. Although the biological activities of Pfs25H and Pfs25M, whether generated by monomeric forms or conjugated nanoparticles, appeared similar, fine-mapping studies with two transmission-blocking monoclonal antibodies detected structural and immunological differences. In addition, evaluation of antisera generated against conjugated Pfs25H or Pfs25M nanoparticles in nonhuman primates identified polyclonal IgG that recognized these structural differences.

ABSTRACTThe amino-acid composition of the immunoglobulin variable region has been observed to impact antibody pharmacokinetics (PK). Here, we sought to improve the PK of the broad HIV−1-neutralizing VRC01-class antibodies, VRC07-523LS and N6LS, by reducing the net positive charge in their variable domains. We used a structure-guided approach to generate a panel of antibody variants incorporating select Arg or Lys substituted to Asp, Gln, Glu, or Ser. The engineered variants exhibited reduced affinity to heparin, reduced polyreactivity, and improved PK in human FcRn-transgenic mice. One variant, VRC07-523LS.v34, with three charge substitutions, had an observed in vivo half-life and an estimated human half-life of 10.8 and 60 days, respectively (versus 5.4 and 38 days for VRC07-523LS) and retained functionality, neutralizing 92% of a 208-strain panel at a geometric mean IC80

ABSTRACTAntibody CAP256-VRC26.25 targets the second hypervariable region (V2) at the apex of the HIV envelope (Env) trimer with extraordinary neutralization potency, although less than optimal breadth. To improve breadth, we linked the light chain of CAP256V2LS, an optimized version of CAP256-VRC26.25 currently under clinical evaluation, to the llama nanobody J3, which has broad CD4-binding site-directed neutralization. The J3-linked bispecific antibody exhibited improved breadth and potency over both J3 and CAP256V2LS, indicative of synergistic neutralization. The cryo-EM structure of the bispecific antibody in complex with a prefusion-closed Env trimer revealed simultaneous binding of J3 and CAP256V2LS. We further optimized the pharmacokinetics of the bispecific antibody by reducing the net positive charge of J3. The optimized bispecific antibody, which we named CAP256.J3LS, had a half-life similar to CAP256V2LS in human FcRn knock-in mice and exhibited suitable auto-reactivity, manufacturability, and biophysical risk. CAP256.J3LS neutralized over 97% of a multiclade 208-strain panel (geometric mean concentration for 80% inhibition (IC80) 0.079 μg/ml) and 100% of a 100-virus clade C panel (geometric mean IC80 of 0.05 μg/ml), suggesting its anti-HIV utility especially in regions where clade C dominates.

Kombucha is a functional beverage made from sugared tea fermented by yeasts, acetic acid bacteria and lactic acid bacteria. In recent years, with the improvement of people's health awareness, Kombucha with health benefits has received widespread attention from all over the world. In this review, the chemical composition of Kombucha, including organic acids, tea polyphenols and aromatic substances, and the sensory quality, including appearance, aroma and taste are summarized. The effects of different fermentation parameters on the chemical composition and sensory quality of Kombucha are discussed. Besides, the interactions between those two are drawn. The information gathered in this review could help us to better understand the sensory quality of Kombucha and the biochemical causes of its formation. Furthermore, it would provide a theoretical basis to the product stability, quality improvement and variety innovation of Kombucha.

Abstract The broadly neutralizing antibody (bNAb) CAP256-VRC26.25 has exceptional potency against HIV-1 and has been considered for clinical use. During the characterization and production of this bNAb, we observed several unusual features. First, the antibody appeared to adhere to pipette tips, requiring tips to be changed during serial dilution to accurately measure potency. Second, during production scale-up, proteolytic cleavage was discovered to target an extended heavy chain loop, which was attributed to a protease in spent medium from 2-week culture. To enable large scale production, we altered the site of cleavage via a single amino acid change, K100mA. The resultant antibody retained potency and breadth while avoiding protease cleavage. We also added the half-life extending mutation LS, which improved the in vivo persistence in animal models, but did not impact neutralization activity; we observed the same preservation of neutralization for bNAbs VRC01, N6, and PGDM1400 with LS on a 208-virus panel. The final engineered antibody, CAP256V2LS, retained the extraordinary neutralization potency of the parental antibody, had a favorable pharmacokinetic profile in animal models, and was negative in in vitro assessment of autoreactivity. CAP256V2LS has the requisite potency, developability and suitability for scale-up, allowing its advancement as a clinical candidate.

The Pneumoviridae family of viruses includes human metapneumovirus (HMPV) and respiratory syncytial virus (RSV). The closely related Paramyxoviridae family includes parainfluenza viruses (PIVs). These three viral pathogens cause acute respiratory tract infections with substantial disease burden in the young, the elderly, and the immune-compromised. While promising subunit vaccines are being developed with prefusion-stabilized forms of the fusion glycoproteins (Fs) of RSV and PIVs, for which neutralizing titers elicited by the prefusion (pre-F) conformation of F are much higher than for the postfusion (post-F) conformation, with HMPV, pre-F and post-F immunogens described thus far elicit similar neutralizing responses, and it has been unclear which conformation, pre-F or post-F, would be the most effective HMPV F-vaccine immunogen. Here, we investigate the impact of further stabilizing HMPV F in the pre-F state. We replaced the furin-cleavage site with a flexible linker, creating a single chain F that yielded increased amounts of pre-F stabilized trimers, enabling the generation and assessment of F trimers stabilized by multiple disulfide bonds. Introduced prolines could increase both expression yields and antigenic recognition by the pre-F specific antibody, MPE8. The cryo-EM structure of a triple disulfide-stabilized pre-F trimer with the variable region of antibody MPE8 at 3.25-Å resolution confirmed the formation of designed disulfides and provided structural details on the MPE8 interface. Immunogenicity assessments in naïve mice showed the triple disulfide-stabilized pre-F trimer could elicit high titer neutralization, >10-fold higher than elicited by post-F. Immunogenicity assessments in pre-exposed rhesus macaques showed the triple disulfide-stabilized pre-F could recall high neutralizing titers after a single immunization, with little discrimination in the recall response between pre-F and post-F immunogens. However, the triple disulfide-stabilized pre-F adsorbed HMPV-directed responses from commercially available pooled human immunoglobulin more fully than post-F. Collectively, these results suggest single-chain triple disulfide-stabilized pre-F trimers to be promising HMPV-vaccine antigens.

The tricarboxylic acid cycle is an important metabolic route in almost all living bodies, yet current analytical methods for the detection of the relevant tricarboxylic acids (TCAs) in biological samples are still unsatisfactory due to their complex matrix and physicochemical properties. In this work, a sample method based on solid phase extraction (SPE) in conjunction with liquid chromatography-mass spectrometry (LC-MS) analysis has been developed for the simultaneous determination of 7 TCAs, including α-ketoglutaric acid, malic acid, malonic acid, aconitic acid, citric acid, succinic acid and fumaric acid, in mice faeces. By treatment with macroporous styrene chelate resin and solid phase extraction, the matrix in mice feces was selectively and efficiently removed. Under optimized parameters, the limit of quantitation for the TCAs was in the range of 1-50 ng/mL, and the content of the TCAs in mice feces was determined with a recovery range of 76.2 % to 128.4 %. Interestingly, the content of malic acid, malonic acid, succinic acid and fumaric acid in the normal mice's feces was found to be 2-10 times higher than that of obese mice, indicating a new potential method for the diagnosis of obesity.

Summary: Soluble HIV-1-envelope (Env) trimers elicit immune responses that target their solvent-exposed protein bases, the result of removing these trimers from their native membrane-bound context. To assess whether glycosylation could limit these base responses, we introduced sequons encoding potential N-linked glycosylation sites (PNGSs) into base-proximal regions. Expression and antigenic analyses indicated trimers bearing six-introduced PNGSs to have reduced base recognition. Cryo-EM analysis revealed trimers with introduced PNGSs to be prone to disassembly and introduced PNGS to be disordered. Protein-base and glycan-base trimers induced reciprocally symmetric ELISA responses, in which only a small fraction of the antibody response to glycan-base trimers recognized protein-base trimers and vice versa. EM polyclonal epitope mapping revealed glycan-base trimers –even those that were stable biochemically– to elicit antibodies that recognized disassembled trimers. Introduced glycans can thus mask the protein base but their introduction may yield neo-epitopes that dominate the immune response.

Summary: Elicitation of antibodies that neutralize the tier-2 neutralization-resistant isolates that typify HIV-1 transmission has been a long-sought goal. Success with prefusion-stabilized envelope trimers eliciting autologous neutralizing antibodies has been reported in multiple vaccine-test species, though not in humans. To investigate elicitation of HIV-1 neutralizing antibodies in humans, here, we analyze B cells from a phase I clinical trial of the “DS-SOSIP”-stabilized envelope trimer from strain BG505, identifying two antibodies, N751-2C06.01 and N751-2C09.01 (named for donor-lineage.clone), that neutralize the autologous tier-2 strain, BG505. Though derived from distinct lineages, these antibodies form a reproducible antibody class that targets the HIV-1 fusion peptide. Both antibodies are highly strain specific, which we attribute to their partial recognition of a BG505-specific glycan hole and to their binding requirements for a few BG505-specific residues. Prefusion-stabilized envelope trimers can thus elicit autologous tier-2 neutralizing antibodies in humans, with initially identified neutralizing antibodies recognizing the fusion-peptide site of vulnerability.

Summary: Broadly neutralizing antibodies (bNAbs) against HIV can reduce viral transmission in humans, but an effective therapeutic will require unusually high breadth and potency of neutralization. We employ the OSPREY computational protein design software to engineer variants of two apex-directed bNAbs, PGT145 and PG9RSH, resulting in increases in potency of over 100-fold against some viruses. The top designed variants improve neutralization breadth from 39% to 54% at clinically relevant concentrations (IC80

Abstract CAP256V2LS, a broadly neutralizing monoclonal antibody (bNAb), is being pursued as a promising drug for HIV-1 prevention. The total level of tyrosine-O-sulfation, a post-translational modification, was known to play a key role for antibody biological activity. More importantly, here wedescribe for the first time the significance of the tyrosine-O-sulfation proteoforms. We developed a hydrophobic interaction chromatography (HIC) method to separate and quantify different sulfation proteoforms, which led to the direct functionality assessment of tyrosine-sulfated species. The fully sulfated (4-SO3) proteoform demonstrated the highest in vitro relative antigen binding potency and neutralization efficiency against a panel of HIV-1 viruses. Interestingly, highly variable levels of 4-SO3 were produced by different clonal CHO cell lines, which helped the bNAb process development towards production of a highly potent CAP256V2LS clinical product with high 4-SO3 proteoform. This study presents powerful insight for any biotherapeutic protein development where sulfation may play an important role in product efficacy.

Summary: Omicron subvariants continuingly challenge current vaccination strategies. Here, we demonstrate nearly complete escape of the XBB.1.5, CH.1.1, and CA.3.1 variants from neutralizing antibodies stimulated by three doses of mRNA vaccine or by BA.4/5 wave infection, but neutralization is rescued by a BA.5-containing bivalent booster. CH.1.1 and CA.3.1 show strong immune escape from monoclonal antibody S309. Additionally, XBB.1.5, CH.1.1, and CA.3.1 spike proteins exhibit increased fusogenicity and enhanced processing compared with BA.2. Homology modeling reveals the key roles of G252V and F486P in the neutralization resistance of XBB.1.5, with F486P also enhancing receptor binding. Further, K444T/M and L452R in CH.1.1 and CA.3.1 likely drive escape from class II neutralizing antibodies, whereas R346T and G339H mutations could confer the strong neutralization resistance of these two subvariants to S309-like antibodies. Overall, our results support the need for administration of the bivalent mRNA vaccine and continued surveillance of Omicron subvariants.

Due to the effects of noise disturbances and system resilience, the current methods for rolling bearing fault feature extraction and degradation trend estimation can hardly achieve more satisfactory results. To address the above issues, we propose a different method for fault feature extraction and degradation trend estimation. Firstly, we preset the Bayesian inference criterion to evaluate the complexity of the denoised vibration signal. When its complexity reaches a minimum, the noise disturbances are exactly removed. Secondly, we define the system resilience obtained by the Bayesian network as the intrinsic index of the system, which is used to correct the equipment degradation trend obtained by the multivariate status estimation technique. Finally, the effectiveness of the proposed method is verified by the completeness of the extracted fault features and the accuracy of the degradation trend estimation over the whole life cycle of the bearing degradation data.

The receptor-binding domain (RBD) of the SARS-CoV-2 spike is a primary target of neutralizing antibodies and a key component of licensed vaccines. Substantial mutations in RBD, however, enable current variants to escape immunogenicity generated by vaccination with the ancestral (WA1) strain. Here, we produce and assess self-assembling nanoparticles displaying RBDs from WA1 and BA.5 strains by using the SpyTag:SpyCatcher system for coupling. We observed both WA1- and BA.5-RBD nanoparticles to degrade substantially after a few days at 37 °C. Incorporation of nine RBD-stabilizing mutations, however, increased yield ~five-fold and stability such that more than 50% of either the WA1- or BA.5-RBD nanoparticle was retained after one week at 37 °C. Murine immunizations revealed that the stabilized RBD-nanoparticles induced ~100-fold higher autologous neutralization titers than the prefusion-stabilized (S2P) spike at a 2 μg dose. Even at a 25-fold lower dose where S2P-induced neutralization titers were below the detection limit, the stabilized BA.5-RBD nanoparticle induced homologous titers of 12,795 ID50 and heterologous titers against WA1 of 1767 ID50. Assessment against a panel of β-coronavirus variants revealed both the stabilized BA.5-RBD nanoparticle and the stabilized WA1-BA.5-(mosaic)-RBD nanoparticle to elicit much higher neutralization breadth than the stabilized WA1-RBD nanoparticle. The extraordinary titer and high neutralization breadth elicited by stabilized RBD nanoparticles from strain BA.5 make them strong candidates for next-generation COVID-19 vaccines.

In this study, Fu-brick tea (FBT) was used for kombucha preparation. The succession of microbial community structures, changes in physicochemical properties, and the volatiles were investigated during the kombucha fermentation. The sequencing analysis showed that Komagataeibacter was the most predominant bacterium. Aspergillus and Zygosaccharomyces were the dominant fungi before fermentation whereas Zygosaccharomyces and Derkella were the dominant fungi after 3 days of fermentation. The physicochemical analysis revealed that acetic acid, glucuronic acid, and polyphenols increased by 10.22 g/L, 0.08 g/L, and 177.40 mg/L, respectively, by the end of fermentation. The GC-MS analysis showed that a total of 49 volatile compounds were detected during the fermentation. Moreover, there were great differences in volatile components among the kombucha samples with different fermentation times. Furthermore, the relevance among microbial community and volatile compounds was evaluated through correlation network analysis. The results suggested that Komagataeibacter, Aspergillus, Zygosaccharomyces, and Dekkera were closely related to the main volatile compounds of FBT kombucha. The results in this study may provide deep understanding for constructing the microbiota and improving the quality of FBT kombucha.

Here, the authors analyse the distance between the body of an antibody and a protein antigen denoted as the Antibody-Framework-to-Antigen Distance (AFAD) for about 2000 non-redundant antibody-protein antigen complexes in the Protein Data Bank. They observe that antibodies with exceptionally long AFADs were all broad HIV-1-neutralizing antibodies that targeted densely glycosylated regions on the HIV-1-envelope trimer. The connection between long AFAD and dense glycan was further validated by the cryo-EM structure of antibody 2909 recognizing a glycan hole and by glycan shielding analyses based on molecular dynamics simulations.

RIFINs are Plasmodium surface antigens that suppress the immune response by binding inhibitory receptors such as LAIR1. Here, Xu et al. characterize the interaction between RIFIN-variable 2 domain and a LAIR1 domain and identify LAIR1-binding RIFINs in several Plasmodium species.

Selenium (Se) is a trace element that plays a crucial role in metabolism; a lack of selenium reduces the body’s resistance and immunity, as well as causes other physiological problems. In this study, we aim to identify favorable conditions for improving organic selenium production. The functional microbe Monascus purpureus, which is widely used in food production, was employed to optimize selenium-enriched culture conditions, and its growth mode and selenium-enriched features were investigated. Spectrophotometry, inductively coupled plasma optical emission spectrometry (ICP-OES), and HPLC (High-Performance Liquid Chromatography) were used to determine the effects of various doses of sodium selenite on the selenium content, growth, and metabolism of M. purpureus, as well as the conversion rate of organic selenium. The best culture parameters for selenium-rich M. purpureus included 7.5 mg/100 mL of selenium content in the culture medium, a pH value of 6.8, a culture temperature of 30 °C, and a rotation speed of 180 rpm. Under ideal circumstances, the mycelia had a maximum selenium concentration of approximately 239.17 mg/kg, with organic selenium accounting for 93.45%, monacoline K production reaching 70.264 mg/L, and a secondary utilization rate of external selenium of 22.99%. This study revealed a novel biological route—selenium-rich M. purpureus fermentation—for converting inorganic selenium into organic selenium.

FeSiAl flakes were fabricated by vibrating ball milling the FeSiAl ribbons. And the microwave absorption properties of FeSiAl flakes were improved by doping the multi-wall carbon nanotubes (MWCNTs) with different mass concentrations. The results show that the FeSiAl/MWCNT composites exhibit significantly improved microwave absorption performance with advantages of strong and broadband absorption in the L-band and S-band. In particular, the reflection loss (RL) of the FeSiAl/MWCNT2 composite reaches −7.4 dB at 1.0 GHz, whereupon, through the electromagnetic simulation software CST Microwave Studio, FeSiAl/MWCNT2 all-dielectric metamaterial absorbers (ADMMAs) were macroscopically designed, achieving an ultra-wideband absorption (RL ≤ −10 dB) of 14.4 GHz (3.6~18.0 GHz). It is recognized that the standing wavelength resonance and diffraction effect are responsible for absorbing electromagnetic waves, and the broadband absorption is improved via dielectric dispersion; their synergistic effect makes the ADMMAs exhibit good microwave absorption performance. This work provides a useful method for designing microwave absorption materials with broadband absorption.

New vaccine delivery technologies, such as mRNA, have played a critical role in the rapid and efficient control of SARS-CoV-2, helping to end the COVID-19 pandemic. Enveloped virus-like particles (eVLPs) are often more immunogenic than protein subunit immunogens and could be an effective vaccine platform. Here, we investigated whether the genetic delivery of eVLPs could achieve strong immune responses in mice as previously reported with the immunization of in vitro purified eVLPs. We utilized Newcastle disease virus-like particles (NDVLPs) to display SARS-CoV-2 prefusion-stabilized spikes from the WA-1 or Beta variant (S-2P or S-2Pᵦ, respectively) and evaluated neutralizing murine immune responses achieved by a single-gene-transcript DNA construct for the WA-1 or Beta variant (which we named S-2P-NDVLP-1T and S-2Pᵦ-NDVLP-1T, respectively), by multiple-gene-transcript DNA constructs for the Beta variant (S-2Pᵦ-NDVLP-3T), and by a protein subunit–DNA construct for the WA-1 or Beta variant (S-2P-TM or S-2Pᵦ-TM, respectively). The genetic delivery of S-2P-NDVLP-1T or S-2Pᵦ-NDVLP-1T yielded modest neutralizing responses after a single immunization and high neutralizing responses after a second immunization, comparable to previously reported results in mice immunized with in vitro purified S-2P-NDVLPs. Notably, genetic delivery of S-2Pᵦ-NDVLP-3T yielded significantly higher neutralizing responses in mice after a second immunization than S-2Pᵦ-NDVLP-1T or S-2Pᵦ-TM. Genetic delivery also elicited high spike-specific T-cell responses. Collectively, these results indicate that genetic delivery can provide an effective means to immunize eVLPs and that a multiple-gene transcript eVLP platform may be especially efficacious and inform the design of improved vaccines.

Macroeconomic predicting is a research hotspot in the field of predicting. The accuracy of predicting often directly affects the rationality of decision-making, especially for defense expenditure predicting. This paper studies the residual correction method of prediction model based on time series. Firstly, based on the grey nonlinear Verhulst prediction model, Fourier series is introduced in this paper to correct the residual sequence once and establish a residual correction model. On this basis, this paper also introduces Markov related concepts, creatively introduces the two-dimensional residual data into Markov state transition matrix, classifies it by K-means clustering analysis, and calculates its parameters by PSO algorithm to realize the secondary accurate correction of residual. Finally, a PSO-Markov residual correction method based on Verhulst-Fourier model is proposed. Tested by examples, this method effectively improves the prediction accuracy of the model, and the prediction is more reliable and accurate.

Lemon peel is the major by-product of lemon juice processing and is currently underutilized. In this study, we explored the feasibility of using lemon peel as a raw material for making vinegar. Lemon peel was homogenized, treated with pectinase (30,000 U/g, 0.1%) at 50 °C for 4 h, and then filtered. The obtained lemon peel juice was first subjected to alcoholic fermentation by Saccharomyces cerevisiae var. FX10, and then acetic fermentation by an acid tolerant Acetobacter malorum, OQY-1, which was isolated from the lemon peels. The juice yield of the lemon peel was 62.5%. The alcoholic fermentation yielded a lemon peel wine with an alcoholic content of 5.16%, and the acetic acid fermentation produced a vinegar with a total acid content of 5.04 g/100 mL. A total of 36 volatile compounds were identified from the lemon vinegar, with some compounds such as esters and some alcohols that increased significantly during alcoholic fermentation while alcohols, terpenoids, and some esters decreased significantly during the fermentations. E-nose and E-tongue analyses coupled with principal component and discriminant factor analyses (PCA and DFA) were able to discriminate the samples at different fermentation stages. Overall, this work demonstrates the potential to transform lemon peel into a valuable product, thus reducing the waste of lemon processing and adding value to the industry.

Konjac glucomannan (KGM) oligosaccharides from KGM is a potentially and valuable prebiotics. A KGM degrading strain was screened and identified as Bacillus amyloliquefaciens WX-1. B. amyloliquefaciens WX-1 was shown to efficiently depolymerize KGM and produce a broad range of small molecule oligosaccharides. The strain efficiently processed KGM with 752.9 mg g−1 yields of KGM oligosaccharides (KGOS) at optimal conditions. The KGOS were characterized using TLC, MALDI-TOF-MS and NMR spectroscopy. The result of KGOS composition revealed that it contained a series of oligosaccharides with a degree of polymerization of 2–14 and the disaccharide structures of MM, MG and GM were assumed. The prebiotic function of KGOS to the growth of Lactobacillus was evaluated. This study showed that B. amyloliquefaciens WX-1 provided a basis for the further development as biocatalyst for direct conversion of polysaccharides derived from energy crops as well as agricultural residues to functional prebiotics.

Since the outbreak of the COVID-19 pandemic, widespread infections have allowed SARS-CoV-2 to evolve in human, leading to the emergence of multiple circulating variants. Some of these variants show increased resistance to vaccine-elicited immunity, convalescent plasma, or monoclonal antibodies. In particular, mutations in the SARS-CoV-2 spike have drawn attention. To facilitate the isolation of neutralizing antibodies and the monitoring of vaccine effectiveness against these variants, we designed and produced biotin-labeled molecular probes of variant SARS-CoV-2 spikes and their subdomains, using a structure-based construct design that incorporated an N-terminal purification tag, a specific amino acid sequence for protease cleavage, the variant spike-based region of interest, and a C-terminal sequence targeted by biotin ligase. These probes could be produced by a single step using in-process biotinylation and purification. We characterized the physical properties and antigenicity of these probes, comprising the N-terminal domain (NTD), the receptor-binding domain (RBD), the RBD and subdomain 1 (RBD-SD1), and the prefusion-stabilized spike ectodomain (S2P) with sequences from SARS-CoV-2 variants of concern or of interest, including variants Alpha, Beta, Gamma, Epsilon, Iota, Kappa, Delta, Lambda, Mu, and Omicron. We functionally validated probes by using yeast expressing a panel of nine SARS-CoV-2 spike-binding antibodies and confirmed sorting capabilities of variant probes using yeast displaying libraries of plasma antibodies from COVID-19 convalescent donors. We deposited these constructs to Addgene to enable their dissemination. Overall, this study describes a matrix of SARS-CoV-2 variant molecular probes that allow for assessment of immune responses, identification of serum antibody specificity, and isolation and characterization of neutralizing antibodies.

Ribes L. belongs to the Grossulariaceae family and has important edible, medicinal, ornamental, and landscaping values. Taxonomic classification within this genus is difficult due to its large variety of species, wide distribution, large morphological variations, and presence of two complex taxonomic groups with bisexual or unisexual flowers. Our study aims to clarify the phylogenetic relationships of Ribes L. taxa in China, and further, to provide a reference for a revised global classification of it. The phylogenetic analysis of 52 Ribes L. samples from 30 species was constructed based on restriction site-associated DNA sequencing and single nucleotide polymorphisms. Afterward, two important taxonomic characters were selected for ancestral state reconstruction over the molecular phylogeny. The results showed that the 52 samples could be divided into six branches, i.e., six subgenera, which caused some controversy regarding the morphological classification of Ribes L. in China. The molecular phylogeny supported the separation of subg. Coreosma from subg. Ribesia and subg. Hemibotrya from subg. Berisia and validated the rationale for recognizing subg. Grossularia as an independent subgenus, the rationality of which was further verified by the reconstruction of ancestor traits. Gene flow among Ribes L. was identified and further confirmed our results.

Summary: Recognition of N-linked glycan at residue N276 (glycan276) at the periphery of the CD4-binding site (CD4bs) on the HIV-envelope trimer is a formidable challenge for many CD4bs-directed antibodies. To understand how this glycan can be recognized, here we isolate two lineages of glycan276-dependent CD4bs antibodies. Antibody CH540-VRC40.01 (named for donor-lineage.clone) neutralizes 81% of a panel of 208 diverse strains, while antibody CH314-VRC33.01 neutralizes 45%. Cryo-electron microscopy (cryo-EM) structures of these two antibodies and 179NC75, a previously identified glycan276-dependent CD4bs antibody, in complex with HIV-envelope trimer reveal substantially different modes of glycan276 recognition. Despite these differences, binding of glycan276-dependent antibodies maintains a glycan276 conformation similar to that observed in the absence of glycan276-binding antibodies. By contrast, glycan276-independent CD4bs antibodies, such as VRC01, displace glycan276 upon binding. These results provide a foundation for understanding antibody recognition of glycan276 and suggest its presence may be crucial for priming immunogens seeking to initiate broad CD4bs recognition.

As an essential and challenging technology of fault prediction and health management(PHM), fault prediction technology has been a research focus in the field of fault diagnosis. However, the current model-based fault prediction technology and data-driven fault prediction technology have some limitations, and it is difficult to effectively apply them in practice. Therefore, this paper combines the advantages of two kinds of fault prediction technology, sets the fault distribution function as the membership function of the adaptive fuzzy neural network based on the full analysis of the fault mechanism. The use of the fault distribution function to highly generalize the law of fault occurrence, and the strong self-learning ability of the neural network can effectively tap the potential fault information of the fault data, thereby using the fault distribution function to fit the fault data, and forming a set of membership functions by presetting a variety of membership functions, so as to expand the applicability of the proposed model in fault prediction. The experimental results show that the fault prediction model proposed in this paper has the advantages of high prediction accuracy, fast convergence speed and good applicability.

The cost prediction is an important part of macroeconomic prediction. The fitting degree of the prediction model not only directly influences the prediction accuracy but also determines the effectiveness of the information provided to the decision maker, especially in the defense economy area. This paper uses the model improving method of reverse prediction and makes the best of the advantage of the Verhulst model of reverse prediction which can solve the problem of ‘small sample, uncertainty’ for the prediction of the defense expenditure. Based on the establishment of a reverse prediction Verhulst model which corrects the initial value, the BP neural network and the combined prediction model based on the BP neural network and improved Verhulst model is further established from the residual sequence dimension with the introduction of the BP neural network, and the analysis validation is conducted through the comparison to the Verhulst-BP model established based on the residual sequence of the traditional Verhulst model. China defense expenditure data collected is tested by practice, showing that this combined prediction model can improve the prediction accuracy greatly.

Background: Respiratory syncytial virus (RSV) subtypes, A and B, co-circulate in annual epidemics and alternate in dominance. We have shown that a subtype A RSV fusion (F) glycoprotein, stabilized in its prefusion conformation by DS-Cav1 mutations, is a promising RSV-vaccine immunogen, capable of boosting RSV-neutralizing titers in healthy adults. In both humans and vaccine-tested animals, neutralizing titers elicited by this subtype A DS-Cav1 immunogen were ~ 2- to 3-fold higher against the homologous subtype A virus than against the heterologous subtype B virus. Methods: To understand the molecular basis for this subtype difference, we introduced DS-Cav1 mutations into RSV strain B18537 F, determined the trimeric crystal structure, and carried out immunogenicity studies. Results: The B18537 DS-Cav1 F structure at 2-Å resolution afforded a precise delineation of prefusion F characteristics, including those of antigenic site Ø, a key trimer-apex site. Structural comparison with the subtype A prefusion F indicated 11% of surface residues to be different, with an alpha-carbon root-mean-square deviation (RMSD) of 1.2 Å; antigenic site Ø, however, resulted in 23% of its surface residues and had an alpha-carbon RMSD of 2.2 Å. Immunization of vaccine-tested animals with DS-Cav1-stabilized B18537 F induced neutralizing responses ~100-fold higher than with postfusion B18537 F. Notably, elicited responses neutralized RSV subtypes A and B at similar levels and were directed towards both conserved equatorial and diverse apical regions. Conclusion: We propose that structural differences in apical and equatorial sites–coupled to differently focused immune responses–provide a molecular explanation for observed differences in elicited subtype A and B neutralizing responses. Keywords: antigenic site; crystal structure; epitope; fusion glycoprotein; immunogenicity; neutralization; RSV subtype; vaccine