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HIV-1 infection is initiated by the interaction between the gp120 subunit in the envelope (Env) trimer and the cellular receptor CD4 on host cells. This interaction induces substantial structural rearrangement of the Env trimer. Currently, static structural information for prefusion-closed trimers, CD4-bound prefusion-open trimers, and various antibody-bound trimers is available. However, dynamic features between these static states (e.g., transition structures) are not well understood. Here, we investigate the full transition pathway of a site specifically glycosylated Env trimer between prefusion-closed and CD4-bound-open conformations by collective molecular dynamics and single-molecule Förster resonance energy transfer (smFRET). Our investigations reveal and confirm important features of the transition pathway, including movement of variable loops to generate a glycan hole at the trimer apex and formation or rearrangements of α-helices and β-strands. Notably, by comparing the transition pathway to known Env-structures, we uncover evidence for a transition intermediate, with four antibodies, Ab1303, Ab1573, b12, and DH851.3, recognizing this intermediate. Each of these four antibodies induce population shifts of Env to occupy a newly observed smFRET state: the "occluded-intermediate" state. We propose this occluded-intermediate state to be both a prevalent state of Env and an on-path conformation between prefusion-closed and CD4-bound-open states, previously overlooked in smFRET analyses., Competing Interests: Competing Interests All authors declare no competing interests.

Introduction: Obesity, as indicated by elevated Body Mass Index (BMI), is a well-established global health concern associated with increased morbidity and mortality across diverse populations. However, the influence of BMI on individuals in Agriculture, Forestry, and Fishing (AFF) occupations, characterized by unique challenges and environmental factors, has received limited research attention., Methods: Our study, a prospective cohort analysis, utilized National Health and Nutrition Examination Survey (NHANES) data from 1999-2014, targeting adults above 18 in AFF occupations with comprehensive BMI data, omitting individuals with a history of cancer. Mortality outcomes were extracted from the NHANES mortality file, and BMI was segmented into eight categories. Essential covariates such as age, sex, race, and various health factors were incorporated. The statistical analysis encompassed Cox regression, generalized additive models, smooth curve fitting, and stratified analyses., Results: During 1,005 person-years with 201 all-cause and 57 CVD deaths, we observed L-shaped and U-shaped correlations of BMI with all-cause and CVD mortality, featuring a pivotal inflection at 26.69 and 27.40 kg/m2. Above this BMI threshold of 26.69 and 27.4 kg/m2, all-cause mortality association was not significant while CVD mortality was positive., Conclusions: This study highlights a unique BMI-mortality association in AFF occupations, diverging from standard patterns. The rigorous labor and environmental conditions in AFF jobs suggest that a certain range of higher BMI could reduce mortality risk. This highlights the necessity for tailored health guidelines in different occupations. Future research should concentrate on diverse health indicators and enhanced risk assessment for physically strenuous occupations., Competing Interests: All authors declare that the study was conducted in the absence of any conflict of interest., (Copyright: © 2024 Qi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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 < 0.0001). The PALOMA-3 study analysed 520 patients with MBC. Within the 153 HER2-0 patients, the palbociclib-fulvestrant group showed a significantly longer PFS than the placebo-fulvestrant group (hazard ratio = 0.54, 95% CI 0.30-0.95, p = 0.034). Among the 367 HER2-low-positive patients, palbociclib-fulvestrant improved PFS (hazard ratio = 0.39, 95% CI 0.28-0.54, p < 0.0001)., Interpretation: The combination of a CDK4/6 inhibitor with ET significantly improved PFS in HER2-low-positive patients, while for HER2-0 patients, benefits were primarily observed in patients who had progressed on previous ET. Furthermore, HER2-0 patients may derive limited benefits from first-line CDK4/6 inhibitor treatment. Further work is needed to validate these findings and to delineate patient subsets that are most likely to benefit from the combination of CDK4/6 inhibitors and ET as first-line treatments., Funding: None., Competing Interests: Declaration of interests JKP was the recipient of research funding by the Color Foundation (PI: Plichta). She serves on the National Comprehensive Cancer Network (NCCN) Breast Cancer Screening Committee. Both are unrelated to this work. She also receives funding from the National Institutes of Health (PI: Amundsen; K12HD043446). SMT holds the consulting or advisory role for Novartis, Pfizer, Merck, Eli Lilly, AstraZeneca, Genentech/Roche, Eisai, Sanofi, Bristol Myers Squibb, Seattle Genetics, CytomX Therapeutics, Daiichi Sankyo, Gilead, Ellipses Pharma, 4D Pharma, OncoSec Medical Inc., BeyondSpring Pharmaceuticals, OncXerna, Zymeworks, Zentalis, Blueprint Medicines, Reveal Genomics, ARC Therapeutics, Infinity Therapeutics, Myovant, Zetagen, Umoja Biopharma, Menarini/Stemline, Aadi Biopharma, Bayer, which are unrelated to this work., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

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 IC 80 s 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., (© 2024 The Authors. Advanced Science published by Wiley‐VCH GmbH. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a plasma protein that controls cholesterol homeostasis. Here, we design a human PCSK9 mimic, named HIT01, with no consecutive 9-residue stretch in common with any human protein as a potential heart attack vaccine. Murine immunizations with HIT01 reduce low-density lipoprotein (LDL) and cholesterol levels by 40% and 30%, respectively. Immunization of cynomolgus macaques with HIT01-K21Q-R218E, a cleavage-resistant variant, elicits high-titer PCSK9-directed antibody responses and significantly reduces serum levels of cholesterol 2 weeks after each immunization. However, HIT01-K21Q-R218E immunizations also increase serum PCSK9 levels by up to 5-fold, likely due to PCSK9-binding antibodies altering the half-life of PCSK9. While vaccination with a PCSK9 mimic can induce antibodies that block interactions of PCSK9 with the LDL receptor, PCSK9-binding antibodies appear to alter homeostatic levels of PCSK9, thereby confounding its vaccine impact. Our results nevertheless suggest a mechanism for increasing the half-life of soluble regulatory factors by vaccination., Competing Interests: Declaration of interests B.Z., G.-Y.C., W.-P.K., Y.W., C.W.C., J.R.M., L.O., T.Z., Y.Y., T.L., and P.D.K. are inventors on a United States provisional patent application submitted by the NIH describing PCSK9-mimicking vaccine immunogens., (Published by Elsevier Inc.)

Biological treatment is commonly used in coking wastewater (CWW) treatment. Prokaryotic microbial communities in CWW treatment have been comprehensively studied. However, viruses, as the critical microorganisms affecting microbial processes and thus engineering parameters, still remain poorly understood in CWW treatment context. Employing viromics sequencing, the composition and function of the viral community in CWW treatment were discovered, revealing novel viral communities and key auxiliary metabolic functions. Caudovirales appeared to be the predominant viral order in the oxic-hydrolytic-oxic (OHO) CWW treatment combination, showing relative abundances of 62.47 %, 56.64 % and 92.20 % in bioreactors O1, H and O2, respectively. At the family level, Myoviridae, Podoviridae and Siphoviridae mainly prevailed in bioreactors O1 and H while Phycodnaviridae dominated in O2. A total of 56.23-92.24% of novel viral contigs defied family-level characterization in this distinct CWW habitat. The virus-host prediction results revealed most viruses infecting the specific functional taxa Pseudomonas, Acidovorax and Thauera in the entire OHO combination, demonstrating the viruses affecting bacterial physiology and pollutants removal from CWW. Viral auxiliary metabolic genes (AMGs) were screened, revealing their involvement in the metabolism of contaminants and toxicity tolerance. In the bioreactor O1, AMGs were enriched in detoxification and phosphorus ingestion, where glutathione S-transferase (GSTs) and beta-ketoadipyl CoA thiolase (fadA) participated in biodegradation of polycyclic aromatic hydrocarbons and phenols, respectively. In the bioreactors H and O2, the AMGs focused on cell division and epicyte formation of the hosts, where GDPmannose 4,6-dehydratase (gmd) related to lipopolysaccharides biosynthesis was considered to play an important role in the growth of nitrifiers. The diversities of viruses and AMGs decreased along the CWW treatment process, pointing to a reinforced virus-host adaptive strategy in stressful operation environments. In this study, the symbiotic virus-bacteria interaction patterns were proposed with a theoretical basis for promoting CWW biological treatment efficiency. The findings filled the gaps in the virus-bacteria interactions at the full-scale CWW treatment and provided great value for understanding the mechanism of biological toxicity and sludge activity in industrial wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Broadly neutralizing antibodies targeting the V2 apex of the HIV-1 envelope trimer are among the most common specificities elicited in HIV-1-infected humans and simian-human immunodeficiency virus (SHIV)-infected macaques. To gain insight into the prevalent induction of these antibodies, we isolated and characterized 11 V2 apex-directed neutralizing antibody lineages from SHIV-infected rhesus macaques. Remarkably, all SHIV-induced V2 apex lineages were derived from reading frame two of the rhesus DH3-15*01 gene. Cryo-EM structures of envelope trimers in complex with antibodies from nine rhesus lineages revealed modes of recognition that mimicked three canonical human V2 apex-recognition modes. Notably, amino acids encoded by DH3-15*01 played divergent structural roles, inserting into a hole at the trimer apex, H-bonding to an exposed strand, or forming part of a loop scaffold. Overall, we identify a DH3-15*01-signature for rhesus V2 apex broadly neutralizing antibodies and show that highly selected genetic elements can play multiple roles in antigen recognition., Highlights: Isolated 11 V2 apex-targeted HIV-neutralizing lineages from 10 SHIV-infected Indian-origin rhesus macaquesCryo-EM structures of Fab-Env complexes for nine rhesus lineages reveal modes of recognition that mimic three modes of human V2 apex antibody recognitionAll SHIV-elicited V2 apex lineages, including two others previously published, derive from the same DH3-15*01 gene utilizing reading frame twoThe DH3-15*01 gene in reading frame two provides a necessary, but not sufficient, signature for V2 apex-directed broadly neutralizing antibodiesStructural roles played by DH3-15*01-encoded amino acids differed substantially in different lineages, even for those with the same recognition modePropose that the anionic, aromatic, and extended character of DH3-15*01 in reading frame two provides a selective advantage for V2 apex recognition compared to B cells derived from other D genes in the naïve rhesus repertoireDemonstrate that highly selected genetic elements can play multiple roles in antigen recognition, providing a structural means to enhance recognition diversity.

High-energy-density lithium-metal batteries (LMBs) coupling lithium-metal anodes and high-voltage cathodes are hindered by unstable electrode/electrolyte interphases (EEIs), which calls for the rational design of efficient additives. Herein, we analyze the effect of electron structure on the coordination ability and energy levels of the additive, from the aspects of intramolecular electron cloud density and electron delocalization, to reveal its mechanism on solvation structure, redox stability, as-formed EEI chemistry, and electrochemical performances. Furthermore, we propose an electron reconfiguration strategy for molecular engineering of additives, by taking sorbide nitrate (SN) additive as an example. The lone pair electron-rich group enables strong interaction with the Li ion to regulate solvation structure, and intramolecular electron delocalization yields further positive synergistic effects. The strong electron-withdrawing nitrate moiety decreases the electron cloud density of the ether-based backbone, improving the overall oxidation stability and cathode compatibility, anchoring it as a reliable cathode/electrolyte interface (CEI) framework for cathode integrity. In turn, the electron-donating bicyclic-ring-ether backbone breaks the inherent resonance structure of nitrate, facilitating its reducibility to form a N-contained and inorganic Li 2 O-rich solid electrolyte interface (SEI) for uniform Li deposition. Optimized physicochemical properties and interfacial biaffinity enable significantly improved electrochemical performance. High rate (10 C), low temperature (-25 °C), and long-term stability (2700 h) are achieved, and a 4.5 Ah level Li||NCM811 multilayer pouch cell under harsh conditions is realized with high energy density (462 W h/kg). The proof of concept of this work highlights that the rational ingenious molecular design based on electron structure regulation represents an energetic strategy to modulate the electrolyte and interphase stability, providing a realistic reference for electrolyte innovations and practical LMBs.

Achieving radar-infrared compatible camouflage with dynamic adaptability has been a long-sought goal, but faces significant challenges owing to the limited dispersion relations of conventional material systems operating in different wavelength ranges. Here, this work proposes the concept of pneumatic multiscale shape morphing and design a periodically arranged pneumatic unit consisting of MXene-based morphable conductors and intake platforms. During gas actuation, the morphable conductor transforms centimeter-scale 2D flat sheets into 3D balloon shapes to enhance microwave absorption behavior, and also reconfigures micrometer-scale MXene wrinkles into smooth planes in combination with cavity-induced low heat transfer to minimize infrared (IR) signatures. Through theory-guided reverse engineering, the final pneumatic matrix shows remarkable frequency tunability (2.64-18.0 GHz), moderate IR emissivity regulation (0.14 at 7-16.5 µm), rapid responsiveness (≈30 ms), wide-angle operation (>45 ° ), and excellent environmental tolerance. Additionally, the multiplexed pneumatic matrix enables over 14 programmable coding sequences that independently alter thermal radiation without compromising radar stealth, and allows multimodal camouflage switching between three distinct compatible states. The approach may facilitate the evolution of camouflage techniques and electromagnetic functional materials toward multispectral, adaptability and intelligence., (© 2024 Wiley‐VCH GmbH.)

Objectives: This study aimed to investigate the association between negative aging stereotypes and goal pursuit in daily life among older adults. We also explored the roles of stereotype threat and stereotype challenge reactions in mediating this association. Additionally, this study investigated whether variations in these associations exist among older adults based on their self-integrity levels., Methods: Participants were 100 older adults who completed daily measures assessing negative aging stereotype experiences, threat and challenge reactions, goal pursuit activities, and self-integrity over a week., Results: More daily experiences of negative aging stereotypes were associated with greater avoidance of responsibilities in goal pursuit and less progress toward goals. Increased threat reactions and decreased challenge reactions were mediators of the association between stereotype experiences and avoidance of responsibilities, as well as that between stereotype experiences and progress toward goals, respectively. Moreover, the associations between threat reactions and avoidance of responsibilities as well as between stereotype experiences and challenge reactions were more pronounced in older adults with lower self-integrity levels., Discussion: This study is pioneering in demonstrating the real-life interplay between aging stereotypes and goal pursuit among older adults. Its findings not only expand upon the literature concerning aging stereotypes, but also offer theoretical insights for the development of interventions aimed at goal pursuit. These insights have significant implications for fostering healthy aging., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Gerontological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

The aim of this study was to investigate the relationships among aging self-stereotype, sense of coherence (SOC), and social isolation, as well as further explore the moderating role of cellphone use in older adults. The 817 Chinese older adults were assessed with regard to aging self-stereotypes, social isolation, SOC and cellphone use. Results from mediation analyses revealed that the negative effect of aging self-stereotype on social isolation was mediated by SOC. Moderated mediation analyses further indicated that the path from SOC to social isolation was stronger and the path from aging self-stereotype to social isolation was weaker for older adults with a higher level of cellphone use. These findings indicated that negative stereotypes were related to weaker SOC and are associated with more severe social isolation. Frequent use of cellphones can alleviate the negative effect of aging self-stereotype on social isolation and enhance the positive effect of SOC on social isolation., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Structural dynamics of human immunodeficiency virus 1 (HIV-1) envelope (Env) glycoprotein mediate cell entry and facilitate immune evasion. Single-molecule FRET using peptides for Env labeling revealed structural dynamics of Env, but peptide use risks potential effects on structural integrity/dynamics. While incorporating noncanonical amino acids (ncAAs) into Env by amber stop-codon suppression, followed by click chemistry, offers a minimally invasive approach, this has proved to be technically challenging for HIV-1. Here, we develope an intact amber-free HIV-1 system that overcomes hurdles of preexisting viral amber codons. We achieved dual-ncAA incorporation into Env on amber-free virions, enabling single-molecule Förster resonance energy transfer (smFRET) studies of click-labeled Env that validated the previous peptide-based labeling approaches by confirming the intrinsic propensity of Env to dynamically sample multiple conformational states. Amber-free click-labeled Env also enabled real-time tracking of single virion internalization and trafficking in cells. Our system thus permits in-virus bioorthogonal labeling of proteins, compatible with studies of virus entry, trafficking, and egress from cells., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Ideal radar absorbing materials (RAMs) require instantaneous, programmable, and spontaneous adaptability to cope with a complex electromagnetic (EM) environment across the full working frequency. Despite various material systems and adaptive mechanisms having been demonstrated, it remains a formidable challenge to integrate these benefits simultaneously. Here, we present a pneumatic matrix that couples morphable MXene/elastomer conductors with dielectric spacers, which leverages controllable airflow to reconfigure the spatial structure between a flat sheet and a hemispherical crown while maintaining resistance stability via wrinkle folding and unfolding. The interdimensional reconfigurations drastically induce multiple resonance behavior, enabling the matrix remarkable frequency tunability (144.5%), ultrawide bandwidth (15 GHz), weak angular dependence (45° incidence), ultrafast responsiveness (∼30 ms), and excellent reproducibility (1000 cycles). With multichannel fluidic and conceptual automated control systems, the final pneumatic device demonstrates a multiplexed, programmable, and autonomous transformable mode that builds a promising platform for smart radar cloaking.

Background: The relationship of serum 25(OH)D levels and hyperlipidemia has not been explored in the Agriculture, Forestry, and Fishing (AFF) occupation. We aimed to explore the impact of serum 25(OH)D levels on lipid profiles in AFF workers, traffic drivers, and miners., Methods: Data from 3937 adults aged 18-65 years old with completed information were obtained from the National Health and Examination Survey from 2001 to 2014. Multivariate linear regression models were used to examine the associations between serum 25(OH)D concentrations and triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and HDL-C/LDL-C ratio. Subgroup analyses for AFF workers considered age, sex, BMI, work activity, months worked, and alcohol consumption. Non-linear relationships were explored using curve fitting., Results: Serum 25(OH)D levels differed between groups (AFF: 60.0 ± 21.3 nmol/L, drivers: 56.6 ± 22.2 nmol/L, miners: 62.8 ± 22.3 nmol/L). Subgroup analysis of the AFF group showed that participants with serum 25(OH)D ≥50 nmol/L, females, and BMI <30 kg/m2 demonstrated improved HDL-C levels correlating with higher serum 25(OH)D. Serum 25(OH)D in AFF workers had a reversed U-shaped relationship with TG and TC, and a U-shaped relationship with HDL-C, with HDL-C, with inflection points at 49.5 nmol/L for TG and TC, and 32.6 nmol/L for HDL-C., Conclusions: Serum 25(OH)D levels are associated with lipid profiles, and the relationship varies among occupational groups. AFF workers, facing unique occupational challenges, may benefit from maintaining adequate serum 25(OH)D levels to mitigate adverse lipid profiles and reduce cardiovascular risk., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Zhang, Dong. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The aims of this study were to explore the predictive relationship between sleep quality and sense of coherence (SOC) and to examine a possible moderating role of mastery in this relationship. A three-wave longitudinal design was employed using a sample of 304 older adults aged 55-87 years old. Cross-lagged panel analyses and moderating effect analyses showed that sleep quality can predict the levels of SOC 6 months later, whereas SOC cannot predict sleep quality 6 months later. In addition, mastery can moderate the effect of sleep quality on SOC. Specifically, the lagged effects of sleep quality on SOC in older adults who had low levels of mastery were stronger than in those who had high levels of mastery. Overall, these findings provide valuable insights for understanding the predictive relationship between sleep quality and SOC and emphasise the moderating role of mastery. Also, our results offer important implications for enhancing the SOC in older adults by improving sleep quality and mastery., (© 2023 John Wiley & Sons Ltd.)

The fusion peptide (FP) on the HIV-1 envelope (Env) trimer can be targeted by broadly neutralizing antibodies (bNAbs). Here, we evaluated the ability of a human FP-directed bNAb, VRC34.01, along with two vaccine-elicited anti-FP rhesus macaque mAbs, DFPH-a.15 and DF1W-a.01, to protect against simian-HIV (SHIV) BG505 challenge. VRC34.01 neutralized SHIV BG505 with a 50% inhibitory concentration (IC 50 ) of 0.58 μg/ml, whereas DF1W-a.01 and DFPH-a.15 were 4- or 30-fold less potent, respectively. VRC34.01 was infused into four rhesus macaques at a dose of 10 mg/kg and four rhesus macaques at a dose of 2.5 mg/kg. The animals were intrarectally challenged 5 days later with SHIV BG505 . In comparison with all 12 control animals that became infected, all four animals infused with VRC34.01 (10 mg/kg) and three out of four animals infused with VRC34.01 (2.5 mg/kg) remained uninfected. Because of the lower potency of DF1W-a.01 and DFPH-a.15 against SHIV BG505 , we infused both Abs at a higher dose of 100 mg/kg into four rhesus macaques each, followed by SHIV BG505 challenge 5 days later. Three of four animals that received DF1W-a.01 were protected against infection, whereas all animals that received DFPH-a.15 were protected. Overall, the protective serum neutralization titers observed in these animals were similar to what has been observed for other bNAbs in similar SHIV infection models and in human clinical trials. In conclusion, FP-directed mAbs can thus provide dose-dependent in vivo protection against mucosal SHIV challenges, supporting the development of prophylactic vaccines targeting the HIV-1 Env FP.

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., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Background: The adaptation of students to academic challenges in high school is crucial for academic performance. This study proposes the concept of "learning crafting," a previously under-researched area, and investigates its associated variables., Methods: Using a diary method, we studied 187 Chinese high school students (64% female; M age  = 15.57) over a 9-day period. We examined the effect of daily life events on learning crafting, and considered academic emotions as mediators and regulatory focus as moderators., Results: Hierarchical linear modeling revealed that daily positive events were positively correlated with learning crafting at both within-person and between-person levels. Positive academic emotions served as mediators of this relationship. Furthermore, promotion focus had a positive moderating effect on the relationship between daily positive events and positive academic emotions. Conversely, daily negative events were only negatively correlated with learning crafting at the between-person level, and no additional significant relationships were identified., Conclusion: This study elucidated the effect of daily life events on learning crafting, its mediating mechanisms, and conditional factors. These results not only contribute to crafting theory, but also provide theoretical underpinnings for future interventions targeting high school students' learning crafting., (© 2023 Foundation for Professionals in Services to Adolescents.)

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 ID 50 and heterologous titers against WA1 of 1767 ID 50 . 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.

Lithium-sulfur batteries (LSBs) are one of the most promising candidates for next-generation energy storage systems. To develop long-life LSBs, there is an urgent need to develop functional materials with higher catalytic activity toward polysulfides and reduced dendritic lithium growth. Herein, an electrostatic field electrocatalyst is designed in a zwitterionic covalent organic framework (COF) with a "two birds with one stone" ability for simultaneously overcoming obstacles in the lithium metal anode and sulfur cathode. The synergism between cationic and anionic moieties in the zwitterionic COF creates an electrostatic field for bidirectionally catalyzing S cathode conversion. Besides, the rational design of zwitterionic COF as a separator modification layer allows lithium bis(trifluoromethanesulfonyl)imide (LiTFSI) dissociation and fast lithium-ion conduction, which alleviates lithium dendrite growth and thus improves the cycling life of LSBs. This contribution not only pioneers the application of zwitterionic COF in the field of LSBs but also highlights the potential of electrostatic field electrocatalysts.

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.

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&#95;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., (© 2023. The Author(s).)

Older adults may be confronted with a variety of negative aging stereotypes (e.g., "forgetful," "physically frail," and "lonely") almost every day. While experimental studies have demonstrated the impact of negative aging stereotypes on older adults' cognitive performance, the relationship between multiple negative aging stereotype experiences and cognitive functioning in older people's daily lives is largely unknown. Using a 1-week daily diary study approach, the present studies examined the association between experiencing daily negative aging stereotypes and real-life cognitive functioning and the mediating role of daily negative affect. We also examined whether these associations differed in response to negative aging stereotype diversity, which refers to the breadth and evenness of negative aging stereotypes that older adults experience in 1 week. In Study 1 ( n = 50), we explored the negative aging stereotypes that older adults often experience in their daily lives. These stereotypes can be seen as a measure of daily stereotype experiences. Then in Study 2, 100 older adults reported their everyday experiences of negative aging stereotypes, positive or negative affect, and cognitive failures for 1 week. The results showed that the more negative aging stereotype experiences older adults had in 1 day, the more cognitive failures occurred that day and that daily negative affect mediated this association. Further, the direct and indirect relationships between daily negative aging stereotype experiences and daily cognitive failures were significant in older adults with high negative aging stereotype diversity but not in those with low negative aging stereotype diversity. This study provides new insights for studies on aging stereotypes threat and offers theoretical guidance for future interventions for cognitive health in older adults and, in turn, contributes to promoting healthy aging in older adults. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

The merging of traditional microwave absorbing materials with metamaterials holds significant potential for enhancing microwave absorber performance. To unlock this potential, a unified paradigm is urgently required. We have successfully established such a paradigm, focusing on regulating effective electromagnetic parameters and interfacial forms across microscopic, mesoscopic, and macroscopic scales. Building upon this foundation, we introduce an active co-design methodology for jointly optimizing full-scale structures and the concept of "full-scale microwave absorbers" (FSMAs). Under this guidance, performance improvements can be achieved efficiently, leading to crucial breakthroughs. For demonstration, we present a case study designing ultra-thin miniaturized FSMAs capable of ultra-broadband and low-frequency absorption. Simulation results show absorptivity exceeding 90% in the 2-28 GHz range, with absorptivity surpassing 85% and 74% in the 1.5-2 GHz and 1-1.5 GHz ranges, respectively. Additionally, the total thickness and macro period are only 5 mm, roughly equivalent to 0.033 wavelengths of the lowest operating frequency. Most importantly, we have broken the Rozanov limit, with experimental results further validating this design. This work significantly enhances our understanding of microwave absorption and offers a shortcut for pursuing improved performances and breakthroughs.

The aim of the current study was to examine the association between body image and hopelessness as well as the roles of aging self-stereotypes and marital status in this association in older adults. There were 821 older adults who participated in the present study and completed questionaries about body image, aging self-stereotypes, hopelessness, demographic information (age and sex), marital status, and health status. The results showed that body image was associated with hopelessness in older adults, and aging self-stereotypes mediated the link between body image and hopelessness. Moderated analyses further indicated that the path from body image to aging self-stereotypes was stronger for single older adults than for those who were married. The results emphasize that older adults' dissatisfaction with their body image can enhance negative aging self-stereotypes, which then result in more severe hopelessness. Marital relationships can alleviate the negative effect of body image on aging self-stereotypes in older adults., (© 2023 Institute of Psychology, Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

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., Competing Interests: LO, BZ, TZ, WPK, YT, JRM, UJB, and PDK are inventors on a U.S. Patent Application filed on their behalf by the National Institutes of Health. The other authors declare no competing interests., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

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.

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.

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., (© 2023. Springer Nature Limited.)

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 (IC 80  < 1 μg/mL) and improve median potency (IC 80 ) by up to 4-fold over a cross-clade panel of 208 strains. To investigate the mechanisms of improvement, we determine cryoelectron microscopy structures of each variant in complex with the HIV envelope trimer. Surprisingly, we find the largest increases in breadth to be a result of optimizing side-chain interactions with highly variable epitope residues. These results provide insight into mechanisms of neutralization breadth and inform strategies for antibody design and improvement., Competing Interests: Declaration of interests B.R.D. and M.S.F. are founders of Ten63 Therapeutics. B.R.D., S.W., A.U.L., G.T.H., M.S.F., P.D.K., J.G., and N.A.D. are inventors on a patent application filed by Duke University., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

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., Competing Interests: Declaration of interests The authors declare no competing interests., (Published by Elsevier Inc.)

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., Competing Interests: NIH has submitted a PCG patent application (PCT/US2023/065009) for Env trimers described in this manuscript on which A.S.O., C.C., T.Z., D.H.R., A.C., R.R., Y.Y., and P.D.K. are co-inventors. The other authors declare no competing interests.

Mechano-optical systems with on-demand adaptability and a broad spectrum from the visible to microwave are critical for complex multiband electromagnetic (EM) applications. Most existing material systems merely have dynamic optical or microwave tunability because their EM wave response is strongly wavelength-dependent. Inspired by cephalopod skin, we develop an adaptive multispectral mechano-optical system based on bilayer acrylic dielectric elastomer (ADE)/silver nanowire (AgNW) films, which reconfigures the surface morphology between wrinkles and cracks via mechanical contraction and stretching. Such morphological evolution regulates the direct transmission/reflection and scattering behavior of visible-infrared light and simultaneously alters the conductive network in a AgNW film to influence its microwave characteristics. The designed system features switching between visible-infrared-microwave transparency and opacity, continuous regulation, wide spectral window (0.38-15.5 μm and 24,200-36,600 μm), excellent recyclability (500 times), and rapid response time (<1 s). These grant the system great potential as platforms for various promising applications such as smart windows, switchable EM devices, dynamic thermal management, adaptive visual stealth, and human motion detection.

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., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Zhang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

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.

Alphaviruses are RNA viruses that represent emerging public health threats. To identify protective antibodies, we immunized macaques with a mixture of western, eastern, and Venezuelan equine encephalitis virus-like particles (VLPs), a regimen that protects against aerosol challenge with all three viruses. Single- and triple-virus-specific antibodies were isolated, and we identified 21 unique binding groups. Cryo-EM structures revealed that broad VLP binding inversely correlated with sequence and conformational variability. One triple-specific antibody, SKT05, bound proximal to the fusion peptide and neutralized all three Env-pseudotyped encephalitic alphaviruses by using different symmetry elements for recognition across VLPs. Neutralization in other assays (e.g., chimeric Sindbis virus) yielded variable results. SKT05 bound backbone atoms of sequence-diverse residues, enabling broad recognition despite sequence variability; accordingly, SKT05 protected mice against Venezuelan equine encephalitis virus, chikungunya virus, and Ross River virus challenges. Thus, a single vaccine-elicited antibody can protect in vivo against a broad range of alphaviruses., Competing Interests: Declaration of interests NIH has submitted a provisional patent application for select antibodies described in this manuscript on which M.S.S., S.K., R.V., P.D.K., and M.R. are co-inventors., (Published by Elsevier Inc.)

While neutralizing antibodies that target the HIV-1 fusion peptide have been elicited in mice by vaccination, antibodies reported thus far have been from only a single antibody class that could neutralize ~30% of HIV-1 strains. To explore the ability of the murine immune system to generate cross-clade neutralizing antibodies and to investigate how higher breadth and potency might be achieved, we tested 17 prime-boost regimens that utilized diverse fusion peptide-carrier conjugates and HIV-1 envelope trimers with different fusion peptides. We observed priming in mice with fusion peptide-carrier conjugates of variable peptide length to elicit higher neutralizing responses, a result we confirmed in guinea pigs. From vaccinated mice, we isolated 21 antibodies, belonging to 4 distinct classes of fusion peptide-directed antibodies capable of cross-clade neutralization. Top antibodies from each class collectively neutralized over 50% of a 208-strain panel. Structural analyses - both X-ray and cryo-EM - revealed each antibody class to recognize a distinct conformation of fusion peptide and to have a binding pocket capable of accommodating diverse fusion peptides. Murine vaccinations can thus elicit diverse neutralizing antibodies, and altering peptide length during prime can improve the elicitation of cross-clade responses targeting the fusion peptide site of HIV-1 vulnerability. IMPORTANCE The HIV-1 fusion peptide has been identified as a site for elicitation of broadly neutralizing antibodies, with prior studies demonstrating that priming with fusion peptide-based immunogens and boosting with soluble envelope (Env) trimers can elicit cross-clade HIV-1-neutralizing responses. To improve the neutralizing breadth and potency of fusion peptide-directed responses, we evaluated vaccine regimens that incorporated diverse fusion peptide-conjugates and Env trimers with variation in fusion peptide length and sequence. We found that variation in peptide length during prime elicits enhanced neutralizing responses in mice and guinea pigs. We identified vaccine-elicited murine monoclonal antibodies from distinct classes capable of cross-clade neutralization and of diverse fusion peptide recognition. Our findings lend insight into improved immunogens and regimens for HIV-1 vaccine development., Competing Interests: The authors declare no conflict of interest.

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., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

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., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Monoclonal antibody L9 recognizes the Plasmodium falciparum circumsporozoite protein (PfCSP) and is highly protective following controlled human malaria challenge. To gain insight into its function, we determined cryoelectron microscopy (cryo-EM) structures of L9 in complex with full-length PfCSP and assessed how this recognition influenced protection by wild-type and mutant L9s. Cryo-EM reconstructions at 3.6- and 3.7-Å resolution revealed L9 to recognize PfCSP as an atypical trimer. Each of the three L9s in the trimer directly recognized an Asn-Pro-Asn-Val (NPNV) tetrapeptide on PfCSP and interacted homotypically to facilitate L9-trimer assembly. We analyzed peptides containing different repeat tetrapeptides for binding to wild-type and mutant L9s to delineate epitope and homotypic components of L9 recognition; we found both components necessary for potent malaria protection. Last, we found the 27-residue stretch recognized by L9 to be highly conserved in P. falciparum isolates, suggesting the newly revealed complete L9 epitope to be an attractive vaccine target., Competing Interests: Declaration of interests L.T.W. and R.A.S. have submitted a US provisional patent application 62/842,590, filed May 3, 2019, describing antibody L9., (Published by Elsevier Ltd.)

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.

Biological treatment processes are an effective method for removing the nitrogen-containing contaminants that exist in coking wastewater. However, little is known about microbial composition and keystone taxa involved in biological nitrogen removal processes. In order to improve the removal efficiency of nitrogen-containing contaminants in anaerobic-aerobic-hydrolytic-aerobic (A/O1/H/O2) system, the microbial composition and interactions of keystone taxa should be clarified. The present work clarifies the removal performance of nitrogen-containing contaminants in the A/O1/H/O2 system, identifies the microbial community involved in various bioreactors, and reveals the keystone taxa within the microbial communities. Combined the processes of ammoniation, denitrification, and nitrification, total nitrogen decreased from 248 to 31 mg L-1 and achieved a removal efficiency of 87.5% in the full-scale A/O1/H/O2 system. High-throughput MiSeq sequencing revealed that Proteobacteria was the most abundant phylum in the A/O1/H/O2 system with relative abundances of 24%-50%. Thiobacillus dominated in bioreactors A and O1 with relative abundances of 2.90% and 4.44%, respectively, while Nitrospira was identified as the most dominant genus in bioreactors H and O2, accounting for 13.33% and 18.38%, respectively. The microbial community composition and co-occurrence network analysis showed that the keystone taxa belonged to Thiobacillus, Nitrospira, Bdellovibrio, Planctomyces, Desulfotomaculum, and Sphingobium, which are related to nitrogen degradation., (© The Author(s) 2022. Published by Oxford University Press on behalf of Applied Microbiology International.)

A phosphorus anode is a promising option for energy-storage applications because of its high theoretical specific capacity and safe lithiation potential. However, the multiphase phosphorus lithiation/delithiation reactions and soluble reaction intermediates cause sluggish reaction kinetics and loss of active materials. Herein, a novel local electric field (LEF) strategy is proposed to inhibit the intermediates dissolution and promote the reaction kinetics by optimizing ionic covalent organic frameworks (iCOFs). Among them, the LEF induced by the cationic covalent organic framework effectively enhances the electrochemical performance of the phosphorus anode. The strong electrostatic interaction between the polyphosphides and cationic covalent organic framework confines the dissolution of active materials and tailors the electronic structure of polyphosphides to accelerate the reaction kinetics. The cationic covalent-organic-framework-assisted phosphorus anode provides a high capacity of 1227.8 mAh g -1 at 10.4 A g -1 (8.6 C) and a high-capacity retention of 87% after 500 cycles at 1.3 A g -1 . This work not only broadens the application of iCOFs for phosphorus anode but also inspires the great potential of the local electric field in battery technology., (© 2022 Wiley-VCH GmbH.)

Antibody 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 (IC 80 ) 0.079 μg/ml) and 100% of a 100-virus clade C panel (geometric mean IC 80 of 0.05 μg/ml), suggesting its anti-HIV utility especially in regions where clade C dominates.

The 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 IC 80 <1 µg/mL. Another variant, N6LS.C49, with two charge substitutions, had an observed in vivo half-life and an estimated human half-life of 14.5 and 80 days (versus 9.0 and 44 days for N6LS) and neutralized ~80% of 208 strains at a geometric mean IC 80 <1 µg/mL. Since Arg and Lys residues are prevalent in human antibodies, we propose substitution of select Arg or Lys with Asp, Gln, Glu, or Ser in the framework region as a general means to improve PK of therapeutic antibodies.

Conjugate-vaccine immunogens require three components: a carrier protein, an antigen, and a crosslinker, capable of coupling antigen to carrier protein, while preserving both T-cell responses from carrier protein and B-cell responses from antigen. We previously showed that the N-terminal eight residues of the HIV-1 fusion peptide (FP8) as an antigen could prime for broad cross-clade neutralizing responses, that recombinant heavy chain of tetanus toxin (rTTHC) as a carrier protein provided optimal responses, and that choice of crosslinker could impact both antigenicity and immunogenicity. Here, we delve more deeply into the impact of varying the linker between FP8 and rTTHC. In specific, we assessed the physical properties, the antigenicity, and the immunogenicity of conjugates for crosslinkers ranging in spacer-arm length from 1.5 to 95.2 Å, with varying hydrophobicity and crosslinking-functional groups. Conjugates coupled with different degrees of multimerization and peptide-to-rTTHC stoichiometry, but all were well recognized by HIV-fusion-peptide-directed antibodies VRC34.01, VRC34.05, PGT151, and ACS202 except for the conjugate with the longest linker (24-PEGylated SMCC; SM(PEG)24), which had lower affinity for ACS202, as did the conjugate with the shortest linker (succinimidyl iodoacetate; SIA), which also had the lowest peptide-to-rTTHC stoichiometry. Murine immunizations testing seven FP8-rTTHC conjugates elicited fusion-peptide-directed antibody responses, with SIA- and SM(PEG)24-linked conjugates eliciting lower responses than the other five conjugates. After boosting with prefusion-closed envelope trimers from strains BG505 clade A and consensus clade C, trimer-directed antibody-binding responses were lower for the SIA-linked conjugate; elicited neutralizing responses were similar, however, though statistically lower for the SM(PEG)24-linked conjugate, when tested against a strain especially sensitive to fusion-peptide-directed responses. Overall, correlation analyses revealed the immunogenicity of FP8-rTTHC conjugates to be negatively impacted by hydrophilicity and extremes of length or low peptide-carrier stoichiometry, but robust to other linker parameters, with several commonly used crosslinkers yielding statistically indistinguishable serological results.

Simian immunodeficiency viruses (SIVs) are lentiviruses that naturally infect non-human primates of African origin and seeded cross-species transmissions of HIV-1 and HIV-2. Here we report prefusion stabilization and cryo-EM structures of soluble envelope (Env) trimers from rhesus macaque SIV (SIV mac ) in complex with neutralizing antibodies. These structures provide residue-level definition for SIV-specific disulfide-bonded variable loops (V1 and V2), which we used to delineate variable-loop coverage of the Env trimer. The defined variable loops enabled us to investigate assembled Env-glycan shields throughout SIV, which we found to comprise both N- and O-linked glycans, the latter emanating from V1 inserts, which bound the O-link-specific lectin jacalin. We also investigated in situ SIV mac -Env trimers on virions, determining cryo-electron tomography structures at subnanometer resolutions for an antibody-bound complex and a ligand-free state. Collectively, these structures define the prefusion-closed structure of the SIV-Env trimer and delineate variable-loop and glycan-shielding mechanisms of immune evasion conserved throughout SIV evolution., (© 2022. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Sustainable production of hydrogen from seawater electrolysis has attracted much attention in recent years. Considering that Cl - might corrode metal substrate by crossing through the covered catalyst, the conventional Ni(II)Fe(III)-layered double hydroxide (NiFe-LDH) loaded on metal substrate, as a favorable oxygen evolution catalyst, cannot be directly used for seawater electrolysis. Herein, an anti-corrosion strategy of PO 4 3- intercalation in NiFe-LDH is proposed, in which the highly negatively charged PO 4 3- in the interlayers can prevent the Ni substrate from Cl - corrosion by electrostatic repulsion. In order to verify the anti-corrosion effect, the two electrodes of the pristine NiFe-LDH and the PO 4 3- intercalated NiFe-LDH are evaluated in a solution with high Cl - concentration. PO 4 3- can effectively hinder the migration of Cl - between the interlayers of NiFe-LDH, thus the corrosion life of the PO 4 3- intercalated NiFe-LDH is more than 100 times longer than that of the pristine NiFe-LDH. The improvement of stability is attributed to the inhibition effect of Cl - passing through the interlayers of NiFe-LDH, leading to the protection of Ni substrate. This work provides a design strategy for the catalysts loaded on the metal substrate, which has excellent Cl - -corrosion resistance and can be widely used in hydrogen generation from seawater electrolysis., (© 2022 Wiley-VCH GmbH.)