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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. [Display omitted] • Designed a human PCSK9 mimic with no consecutive 9 residues of any human protein • Immunization with the PCSK9 mimic significantly reduced cholesterol and LDL levels • Vaccine-induced antibodies altered PCSK9 half-life, confounding vaccine impact Proprotein convertase subtilisin/kexin type 9 (PCSK9) controls cholesterol homeostasis. To assess the feasibility of a PCSK9-based heart attack vaccine, Zhang et al. create a PCSK9 mimic and demonstrate in mice and NHPs its ability to significantly reduce cholesterol. Unexpectedly, the vaccine-induced antibodies also altered the PCSK9 half-life, blunting vaccine impact. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • HIV-1 neutralizing antibodies 2C06 and 2C09 from VRC 018 clinical trial are isolated • Cryo-EM structures of 2C06 and 2C09 in complex with HIV envelope trimer are determined • 2C06 and 2C09 binding requires a glycan hole at 241 and BG505-specific residues • Antibodies 2C06 and 2C09 form a reproducible class, recognizing fusion peptide Vaccine elicitation of tier-2 HIV-1 neutralizing antibodies in humans has been a long-sought goal. Wang et al. isolate and determine structures of two antibodies from a clinical trial of a prefusion-closed HIV-1 envelope trimer. These antibodies target the fusion-peptide site of vulnerability, form a reproducible class, and neutralize the tier-2 HIV-1 strain BG505. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Antibody variants designed using OSPREY improve neutralization breadth and potency • Structural and statistical analyses highlight improved side-chain interactions • Improvements in breadth result from interaction with variable epitope residues Broadly neutralizing antibodies against HIV are promising therapeutics and targets for vaccine elicitation. Using the OSPREY design software, Holt et al. design antibody variants with improved breadth and potency of virus neutralization. They solve bound structures for these variants and provide insight into mechanisms of breadth and potency. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Bivalent booster induces 2- to 8-fold higher nAb titer than monovalent against XBB and XBB.1.5 • CH.1.1 and CA.3.1 exhibit nearly complete escape of neutralization from bivalent booster • XBB.1.5, CH.1.1, and CA.3.1 show increased fusogenicity compared with BA.2 • Homology modeling shows impacts of F486P mutation present in XBB.1.5 on ACE2 binding Qu et al. show that bivalent booster recipients, compared with monovalent recipients, exhibit higher nAb titers against Omicron subvariants XBB, XBB.1, and XBB.1.5. The CH.1.1 and CA.3.1 variants show more substantial neutralization escape than the XBB variants. Further, structural modeling reveals that the F486P mutation in XBB.1.5 enhances ACE2 binding. [ABSTRACT FROM AUTHOR]

Summary While the HIV-1-glycan shield is known to shelter Env from the humoral immune response, its quantitative impact on antibody elicitation has been unclear. Here, we use targeted deglycosylation to measure the impact of the glycan shield on elicitation of antibodies against the CD4 supersite. We engineered diverse Env trimers with select glycans removed proximal to the CD4 supersite, characterized their structures and glycosylation, and immunized guinea pigs and rhesus macaques. Immunizations yielded little neutralization against wild-type viruses but potent CD4-supersite neutralization (titers 1: >1,000,000 against four-glycan-deleted autologous viruses with over 90% breadth against four-glycan-deleted heterologous strains exhibiting tier 2 neutralization character). To a first approximation, the immunogenicity of the glycan-shielded protein surface was negligible, with Env-elicited neutralization (ID 50 ) proportional to the exponential of the protein-surface area accessible to antibody. Based on these high titers and exponential relationship, we propose site-selective deglycosylated trimers as priming immunogens to increase the frequency of site-targeting antibodies. [ABSTRACT FROM AUTHOR]

The emergence and global spread of the SARS-CoV-2 Omicron variants, which carry an unprecedented number of mutations, raise serious concerns due to the reduced efficacy of current vaccines and resistance to therapeutic antibodies. Here, we report the generation and characterization of two potent human monoclonal antibodies, NA8 and NE12, against the receptor-binding domain of the SARS-CoV-2 spike protein. NA8 interacts with a highly conserved region and has a breadth of neutralization with picomolar potency against the Beta variant and the Omicron BA.1 and BA.2 sublineages and nanomolar potency against BA.2.12.1 and BA.4. Combination of NA8 and NE12 retains potent neutralizing activity against the major SARS-CoV-2 variants of concern. Cryo-EM analysis provides the structural basis for the broad and complementary neutralizing activity of these two antibodies. We confirm the in vivo protective and therapeutic efficacies of NA8 and NE12 in the hamster model. These results show that broad and potent human antibodies can overcome the continuous immune escape of evolving SARS-CoV-2 variants. [Display omitted] • Monoclonal antibodies are isolated from COVID-19 convalescent patients • NA8 and NE12 potently neutralize newly emerging SARS-CoV-2 variants of concern • Structural analysis reveals a unique conserved binding epitope for NA8 • NA8 and NE12 show prophylactic and therapeutic efficacy in the Syrian hamster model The emergence of new SARS-CoV-2 variants that escape neutralization can jeopardize the efficacy of vaccines and therapeutic antibodies. Chen et al. report the isolation of potent human antibodies that neutralize emerging variants of concern, including various Omicron sublineages. These antibodies show prophylactic and therapeutic efficacy in the hamster model. [ABSTRACT FROM AUTHOR]

[Display omitted] • Bacillus amyloliquefaciens WX-1 can degrade konjac glucomannan (KGM) for the production of KGM oligosaccharides (KGOS). • The degree of polymerization of KGOS was between 2 and 14 (mainly disaccharide and trisaccharide). • The structural characterization of KGOS was carried out using TLC, MALDI-TOF-MS and NMR. • KGOS produced by B. amyloliquefaciens WX-1 showed good prebiotic activity. 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. [ABSTRACT FROM AUTHOR]

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. [Display omitted] • Glycan276-dependent antibodies with up to 81% neutralization breadth are isolated • Structure for three glycan276-dependent antibodies with envelope trimer is determined • Glycan276-dependent antibodies bind glycan276 with diverse modes of recognition • Glycan276-dependent antibodies maintain ligand-free orientation of glycan276 CD4 binding site-directed antibodies can broadly neutralize circulating HIV strains but often need to overcome glycan shielding, especially from glycan276. Cottrell et al. isolate two lineages of antibodies that depend on glycan276 for neutralization and elucidate the mechanistic difference between glycan276-dependent and glycan276-independent antibodies, with implications for HIV vaccine design. [ABSTRACT FROM AUTHOR]

Lipopeptide containing an ELA-A1-restricted cytotoxic T lymphocyte (CTL) epitope from the envelope surface unit (SU) protein of the EIAVWSU5 strain was used to immunize three horses having the ELA-A1 haplotype. Peptide-specific ELA-A1-restricted CTL were induced in all three horses, although these were present transiently in PBMC. These horses were further immunized with lipopeptide containing the corresponding CTL epitope from the EIAVPV strain. Then, the three immunized horses and three non-immunized horses were challenged by intravenous inoculation with 300 TCID50 EIAVPV. All horses developed cell free viremia, fever and thrombocytopenia. However, there was a statistically lower fever and thrombocytopenia severity score in the immunized group. Shorter duration of plasma viral load in two of the three immunized horses likely explains the less severe clinical disease in this group. Results indicate that lipopeptide immunization had a protective effect against development of clinical disease following virus challenge. [Copyright &y& Elsevier]

Antibodies with heavy chains that derive from the VH1-2 gene constitute some of the most potent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-neutralizing antibodies yet identified. To provide insight into whether these genetic similarities inform common modes of recognition, we determine the structures of the SARS-CoV-2 spike in complex with three VH1-2-derived antibodies: 2-15, 2-43, and H4. All three use VH1-2-encoded motifs to recognize the receptor-binding domain (RBD), with heavy-chain N53I-enhancing binding and light-chain tyrosines recognizing F486 RBD. Despite these similarities, class members bind both RBD-up and -down conformations of the spike, with a subset of antibodies using elongated CDRH3s to recognize glycan N 343 on a neighboring RBD—a quaternary interaction accommodated by an increase in RBD separation of up to 12 Å. The VH1-2 antibody class, thus, uses modular recognition encoded by modular genetic elements to effect potent neutralization, with the VH-gene component specifying recognition of RBD and the CDRH3 component specifying quaternary interactions. [Display omitted] • Determine structures of SARS-CoV-2-neutralizing antibodies 2-43, 2-15, and H4 • Define a multi-donor VH1-2 antibody class for RBD and quaternary recognition • Reveal structural basis of RBD-up and RBD-down recognition within the class • Delineate changes in spike conformation induced by quaternary recognition Rapp et al. determine structures of three human VH1-2-derived SARS-CoV-2-neutralizing antibodies and define a prevalent multi-donor antibody class that recognizes the receptor-binding domain (RBD) with convergent heavy- and light-chain modules. They further show the structural basis for RBD-up and RBD-down recognition and quaternary recognition-mediated spike-conformation change. [ABSTRACT FROM AUTHOR]

Soluble "SOSIP"-stabilized envelope (Env) trimers are promising HIV-vaccine immunogens. However, they induce high-titer responses against the glycan-free trimer base, which is occluded on native virions. To delineate the effect on base responses of priming with immunogens targeting the fusion peptide (FP) site of vulnerability, here, we quantify the prevalence of trimer-base antibody responses in 49 non-human primates immunized with various SOSIP-stabilized Env trimers and FP-carrier conjugates. Trimer-base responses account for ∼90% of the overall trimer response in animals immunized with trimer only, ∼70% in animals immunized with a cocktail of SOSIP trimer and FP conjugate, and ∼30% in animals primed with FP conjugates before trimer immunization. Notably, neutralization breadth in FP-conjugate-primed animals correlates inversely with trimer-base responses. Our data provide methods to quantify the prevalence of trimer-base responses and reveal that FP-conjugate priming, either alone or as part of a cocktail, can reduce the trimer-base response and improve the neutralization outcome. [Display omitted] • Devise methods to quantify antibody responses targeting the base of HIV-1 Env trimers • Fusion-peptide (FP) priming reduces anti-base responses upon HIV Env trimer boost • Lower percentage of anti-base responses correlates with improved neutralization breadth The exposed base region of soluble HIV-1 Env trimers elicits strong non-neutralizing antibody responses. Corrigan et al. quantify plasma anti-base responses in immunized NHPs and observe a reduction in anti-base responses with fusion-peptide priming. The percentage of anti-base responses correlates inversely with neutralization breadth, providing insights for improving vaccination strategies. [ABSTRACT FROM AUTHOR]

Soluble envelope (Env) trimers, stabilized in a prefusion-closed conformation, can elicit neutralizing responses against HIV-1 strains closely related to the immunizing trimer. However, to date such stabilization has succeeded with only a limited number of HIV-1 strains. To address this issue, here we develop ADROITrimer, an automated procedure involving structure-based stabilization and consensus repair, and generate "RnS-DS-SOSIP"-stabilized Envs from 180 diverse Env sequences. The vast majority of these RnS-DS-SOSIP Envs fold into prefusion-closed conformations as judged by antigenic analysis and size exclusion chromatography. Additionally, representative strains from clades AE, B, and C are stabilized in prefusion-closed conformations as shown by negative-stain electron microscopy, and the crystal structure of a clade A strain MI369.A5 Env trimer provides 3.5 Å resolution detail into stabilization and repair mutations. The automated procedure reported herein that yields well-behaved, soluble, prefusion-closed Env trimers from a majority of HIV-1 strains could have substantial impact on the development of an HIV-1 vaccine. • Automated ADROITrimer pipeline enables prefusion-closed stabilization of HIV Env • 134 of 180 RnS-DS-SOSIP Env variants are recognized by PGT145 or CAP256-VRC26.25 • RnS-DS-SOSIP yields soluble Env trimers for 143 of 179 strains as confirmed by SEC • 3.5 Å resolution structure of a clade A RnS-DS-SOSIP reveals stabilization details A general method to stabilize HIV-1 envelope (Env) trimers in its prefusion-closed conformation may have substantial vaccine utility. Rawi et al. report an automated pipeline based on DS-SOSIP and repair-and-stabilization (RnS) strategies, which successfully stabilizes more than 70% of a diverse cross-clade panel of 180 strains in the prefusion-closed conformation. [ABSTRACT FROM AUTHOR]

Biotin-labeled molecular probes, comprising specific regions of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike, would be helpful in the isolation and characterization of antibodies targeting this recently emerged pathogen. Here, we design constructs incorporating an N-terminal purification tag, a site-specific protease-cleavage site, the probe region of interest, and a C-terminal sequence targeted by biotin ligase. Probe regions include full-length spike ectodomain as well as various subregions, and we also design mutants that eliminate recognition of the angiotensin-converting enzyme 2 (ACE2) receptor. Yields of biotin-labeled probes from transient transfection range from ∼0.5 mg/L for the complete ectodomain to >5 mg/L for several subregions. Probes are characterized for antigenicity and ACE2 recognition, and the structure of the spike ectodomain probe is determined by cryoelectron microscopy. We also characterize antibody-binding specificities and cell-sorting capabilities of the biotinylated probes. Altogether, structure-based design coupled to efficient purification and biotinylation processes can thus enable streamlined development of SARS-CoV-2 spike ectodomain probes. • Structure-based design of SARS-CoV-2 spike ectodomain and subdomain probes • On-column biotinylation and purification is enabled by affinity tag and HRV3C cleavage • Development of diverse molecular probes by "cut-and-paste" strategy • Application of probes to SARS-CoV-2 antibody discovery and immune evaluation SARS-CoV-2 spike probes are key to antibody discovery and vaccine evaluation. By using structure-based design, affinity purification, on-column biotinylation, and sequence-specific protease cleavage, Zhou et al. devise a strategy that allows for the rapid development of SARS-CoV-2 spike probes for B cell sorting, antibody discovery, and immune assessment. [ABSTRACT FROM AUTHOR]

Multidonor antibodies are of interest for vaccine design because they can in principle be elicited in the general population by a common set of immunogens. For influenza, multidonor antibodies have been observed against the hemagglutinin (HA) stem, but not the immunodominant HA head. Here, we identify and characterize a multidonor antibody class (LPAF-a class) targeting the HA head. This class exhibits potent viral entry inhibition against H1N1 A/California/04/2009 (CA09) virus. LPAF-a class antibodies derive from the HV2-70 gene and contain a "Tyr-Gly-Asp"-motif, which occludes the HA-sialic acid binding site as revealed by a co-crystal structure with HA. Both germline-reverted and mature LPAF antibodies potently neutralize CA09 virus and have nanomolar affinities for CA09 HA. Moreover, increased frequencies for LPFA-a class antibodies are observed in humans after a single vaccination. Overall, this work highlights the identification of a multidonor class of head-directed influenza-neutralizing antibodies and delineates the mechanism of their recurrent elicitation in humans. • Analysis of HV2-70-derived antibodies reveals basis for a multidonor "LPAF-a" class • LPAF-a.01 antibody inserts extended CDR H3 into influenza sialic acid-binding site • Germline-reverted LPAF-a class antibodies have nanomolar affinity for CA09 antigen • Increased frequency of LPAF-a class antibodies in humans after CA09 vaccination Multidonor influenza-neutralizing antibodies targeting hemagglutinin-head regions may have vaccine utility. Cheung et al. report a HV2-70-derived multidonor class, the "LPAF-a" antibody class, which targets the hemagglutinin head and can be commonly observed in humans after a single vaccination. Recurrent elicitation is attributed to tight binding between hemagglutinin and germline LPAF-a class antibodies. [ABSTRACT FROM AUTHOR]

Antibodies targeting the V1V2 apex of the HIV-1 envelope (Env) trimer comprise one of the most commonly elicited categories of broadly neutralizing antibodies. Structures of these antibodies indicate diverse modes of Env recognition typified by antibodies of the PG9 class and the PGT145 class. The mode of recognition, however, has been unclear for the most potent of the V1V2 apex-targeting antibodies, CAP256-VRC26.25 (named for donor-lineage.clone and referred to hereafter as VRC26.25). Here, we determine the cryoelectron microscopy structure at 3.7 Å resolution of the antigen-binding fragment of VRC26.25 in complex with the Env trimer thought to have initiated the lineage. The 36-residue protruding loop of VRC26.25 displays recognition incorporating both strand-C interactions similar to the PG9 class and V1V2 apex insertion similar to the PGT145 class. Structural elements of separate antibody classes can thus intermingle to form a "combined" class, which in this case yields an antibody of extraordinary potency. • Structure of CAP256-VRC26.25 bound to lineage-initiating Env trimer • CAP256-VRC26.25 engages HIV-1 V1V2 C-strand similarly to the PG9 antibody class • CAP256-VRC26.25 inserts into Env trimer apex similarly to the PGT145 antibody class • VRC26 exhibits "combined" recognition, intermingling components of distinct classes Antibody CAP256-VRC26.25 is one of the most potent known HIV-1-neutralizing antibodies. Gorman et al. determine the cryo-EM structure of this antibody in complex with the Env trimer that initiated the antibody lineage. The structure reveals how elements of distinct antibody classes can intermingle to yield an antibody of extraordinary potency. [ABSTRACT FROM AUTHOR]

Exploiting photocatalysts/catalysts with characteristics of low cost, high reactivity and easy recovery is of great significance for energy efficiency and environmental remediation. Herein, CuO/TiO 2 hybrid nanofibers with different CuO contents were fabricated by a simple and low-cost method of electrospinning followed with calcinations in air. The hybrid nanofibers possess some distinct merits including mesoporous structure, extended light response range, and large TiO 2 /CuO interface. The CuO/TiO 2 hybrid nanofibers showed improved photocatalytic activity for organic dye degradation under both simulated sunlight (UV–vis) and visible light (λ > 420 nm), compared with pure TiO 2 nanofibers. Both processes exhibited humplike activity dependence upon CuO addition, with a peak at 1 wt.%-CuO content under UV–vis light and a peak at 5 wt.%-CuO content under visible light, respectively. The mechanism for the CuO-content dependent photocatalysis was discussed with the photoelectrochemical analysis in evidence. Moreover, the CuO/TiO 2 hybrid nanofibers exhibited very high catalytic efficiency for the reduction of nitrophenols by sodium borohydride, and the apparent reaction rate constants ascended with the increased CuO content. The optimized CuO/TiO 2 nanofibers are expected to act a highly efficient catalyst to replace the noble metals in this field. Therefore, this work demonstrated a convenient case for the exploitation of noble-metal-free catalysts with high efficiency, to meet the increasing demands of industrial applications at reduced costs. [ABSTRACT FROM AUTHOR]

Background: Previous research has shown that perfectionism is closely associated with stress. However, limited research has examined the longitudinal relationship between perfectionism and test anxiety. In addition, no study has investigated the possible mediators for the effects of perfectionism on test anxiety in school contexts.Objective: Utilizing a three-wave longitudinal design, this study employed a sample of Chinese junior high school students as participants to examine the longitudinal relationship between perfectionism, sense of coherence, and test anxiety.Methods: 427 Chinese junior high school students (212 girls, 215 boys) participated in all three measurement times. The Frost Multidimensional Perfectionism scale, sense of coherence scale (SOC), and Test Anxiety scale were administered to the participants. The students completed questionnaires about once every two months from the beginning of a school semester to the end of the semester.Results: Results from structural equation modeling indicated that negative perfectionism significantly contributed to test anxiety in the adolescent students. The lagged effect of perfectionism (T1) on SOC (T2) was significant, and the lagged effect of SOC (T2) on test anxiety (T3) was significant, indicating that SOC played a significant mediating role in the effect of perfectionism on test anxiety.Conclusion: The longitudinal effects of negative perfectionism on test anxiety were mediated by sense of coherence. The findings of the study have general implications for mitigating test anxiety and improving school adjustment and academic achievement. [ABSTRACT FROM AUTHOR]

Using a sample of Chinese teenagers (368 males and 380 females) randomly selected from three middle schools, we integrated masculine gender stereotypes and identity into a structural model of the relationship between fathering style and children’s social problem solving. Results showed that the data had a satisfactory fit to the proposed model (CFI = .96, RMSEA = .030, and SRMR = .0336), indicating that masculine gender stereotype and identity that children hold partially mediated the relationship between fathering style and children’s social problem solving. Moreover, multi-group analyses indicated that the patterns of the mediation model did not differ across gender, except that more masculine stereotypes will lead to more masculine identity for boys than for girls. This finding indicated that positive fathering style can promote positive gender stereotype and identity that children have, which further promote their development of social problem solving. [ABSTRACT FROM AUTHOR]

• The novel viral community involved in CWW treatment system was studied by viromics. • The diversities of viruses and AMGs decreased along the CWW treatment process. • Viral AMGs are involved in contaminants removal and resistance to environmental stresses. • The hosts of viruses were dominantly Pseudomonas, Acidovorax and Thauera genera. • The symbiotic virus-bacteria interaction patterns in CWW treatment were proposed. 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. [Display omitted] [ABSTRACT FROM AUTHOR]

• Parenting differences are risk factors for a low level of psychological resilience in adolescents. • Compared with parenting consistency, parenting differences impede the development of gratitude in adolescents. • Gratitude mediated the relationships between parenting differences and psychological resilience in adolescents. Previous studies demonstrated that parenting differences between fathers and mothers may trigger family conflict, which is a risk factor for failing to develop gratitude and psychological resilience in their offspring. Gratitude has also been shown to be a protective factor for developing psychological resilience. However, little is known about the role of gratitude in parenting differences and psychological resilience. This study examined the effects of parenting differences on gratitude and psychological resilience in adolescents and the mediating role of gratitude on the relationship between parenting differences and psychological resilience. A sample of 1117 Chinese adolescents aged 11–19 years (mean age = 14.32; SD = 1.10) participated in this study and completed measures of parenting, gratitude, and psychological resilience. The results indicated that for the same level of parenting, compared with parenting consistency on a given parenting dimension, parenting differences on that same parenting dimension had more negative effects on gratitude and psychological resilience in adolescents. The results also revealed that gratitude mediated the relationship between parenting differences and psychological resilience in adolescents. These findings can contribute to a better understanding of the inner mechanism of parenting differences affecting psychological resilience of adolescents and provide guidance for effectively improving adolescent individual psychological resilience. [ABSTRACT FROM AUTHOR]

Abstract: ECR-9 (CGS) zeolites have been synthesised from gallosilicate and gallo-aluminosilicate gels at between 100°C and 220°C using several Al and Si reactant types. Aluminum was substituted for gallium in the framework at levels up to 20% in seeded syntheses, but higher Al substitutions promote the crystallization of impurities. This is the first report of a gallo-aluminosilicate CGS composition. We successfully used the Simulated Framework Annealing method to solve the structural topology as if ECR-9 was a novel gallosilicate. The structure was refined using Rietveld full profile analysis (wRp=14.6%) of powder synchrotron X-ray diffraction data. [Copyright &y& Elsevier]

• Purification of clade A and C HIV-1 envelope (Env) trimers by a novel non-affinity method. • Env trimers produced in pre-fusion closed conformation without antibody capture. • Non-affinity purified trimers show minimal exposure of CD4-induced or V3 epitopes. • Scalable process yields up to 0.2 g/L of purified prefusion-closed HIV-1-Env trimer. Metastable glycosylated immunogens present challenges for GMP manufacturing. The HIV-1 envelope (Env) glycoprotein trimer is covered by N -linked glycan comprising half its mass and requires both trimer assembly and subunit cleavage to fold into a prefusion-closed conformation. This conformation, the vaccine-desired antigenic state, is both metastable to structural rearrangement and labile to subunit dissociation. Prior reported GMP manufacturing for a soluble trimer stabilized in a near-native state by disulfide (SOS) and Ile-to-Pro (IP) mutations has employed affinity methods based on antibody 2G12, which recognizes only ~30% of circulating HIV strains. Here, we develop a scalable manufacturing process based on commercially available, non-affinity resins, and we apply the process to current GMP (cGMP) production of trimers from clades A and C, which have been found to boost cross-clade neutralizing responses in vaccine-test species. The clade A trimer, which we named "BG505 DS-SOSIP.664", contained an engineered disulfide (201C-433C; DS) within gp120, which further stabilized this trimer in a prefusion-closed conformation resistant to CD4-induced triggering. BG505 DS-SOSIP.664 was expressed in a CHO-DG44 stable cell line and purified with initial and final tangential flow filtration steps, three commercially available resin-based chromatography steps, and two orthogonal viral clearance steps. The non-affinity purification enabled efficient scale-up, with a 250 L-scale cGMP run yielding 9.6 g of purified BG505 DS-SOSIP.664. Antigenic analysis indicated retention of a prefusion-closed conformation, including recognition by apex-directed and fusion peptide-directed antibodies. The developed manufacturing process was suitable for 50 L-scale production of a second prefusion-stabilized Env trimer vaccine candidate, ConC-FP8v2 RnS-3mut-2G-SOSIP.664, yielding 7.8 g of this consensus clade C trimer. The successful process development and purification scale-up of HIV-1 Env trimers from different clades by using commercially available materials provide experimental demonstration for cGMP manufacturing of trimeric HIV-Env vaccine immunogens, in an antigenically desired conformation, without the use of costly affinity resins. [ABSTRACT FROM AUTHOR]