ABSTRACT
A new type of hybrid material (NCMTs@MoO2/FeNi3) with a multi-layer heterostructure was designed and fabricated via a one-step pyrolysis process using FeOOH/NiMoO4@PDA as the precursor. FeOOH/NiMoO4@PDA was prepared by the solvothermal method, followed by the nickel-ion etching method coupled with the polymerization of dopamine (DA). The as-obtained material was made of nitrogen-doped carbon nanotubes embedded with FeNi3 and MoO2 nanoparticles (NPs). Notably, the FeNi3 NPs exhibited significantly improved performance in the reduction of 4-nitrophenol (4-NP) and adsorption of histidine-rich protein as well as provided appropriate magnetism resources. The MoO2 NPs imparted a metallic nature with excellent conductivity, and the N-doped mesoporous carbon microtubes also improved conductivity and facilitated mass transfer, thus leading to enhanced performance in catalysis. Benefiting from the 1D hierarchical porous structure and compositional features, the NCMTs@MoO2/FeNi3 composites exhibited excellent performance in 4-NP reduction and protein adsorption via specific metal affinity between the polyhistidine groups of proteins and the FeNi3 NPs. The result presented here indicates that the strategy of combining tailored components, heterostructuring, and carbon integration is a promising way to obtain high-performance composites for other energy-related applications.
Subject(s)
Molybdenum , Adsorption , Molybdenum/chemistry , Catalysis , Nanotubes, Carbon/chemistry , Oxides/chemistry , Nickel/chemistry , Nitrophenols/chemistry , Proteins/chemistry , Surface PropertiesABSTRACT
The emerging new variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) needs booster vaccination. We evaluated the long-term safety and immunogenicity of heterologous boosting with a SARS-CoV-2 messenger RNA vaccine SYS6006. A total of 1000 participants aged 18 years or more who had received two (Group A) or three (Group B) doses of SARS-CoV-2 inactivated vaccine were enrolled and vaccinated with one dose of SYS6006 which was designed based on the prototype spike protein and introduced mutation sites. Adverse events (AEs) through 30 days and serious AEs during the study were collected. Live-virus and pseudovirus neutralizing antibody (Nab), binding antibody (immunoglobulin G [IgG]) and cellular immunity were tested through 180 days. Solicited all, injection-site and systemic AEs were reported by 618 (61.8%), 498 (49.8%), and 386 (38.6%) participants, respectively. Most AEs were grade 1. The two groups had similar safety profile. No vaccination-related SAEs were reported. Robust wild-type (WT) live-virus Nab response was elicited with peak geometric mean titers (GMTs) of 3769.5 (Group A) and 5994.7 (Group B) on day 14, corresponding to 1602.5- and 290.8-fold increase versus baseline, respectively. The BA.5 live-virus Nab GMTs were 87.7 (Group A) and 93.2 (Group B) on day 14. All participants seroconverted for WT live-virus Nab. Robust pseudovirus Nab and IgG responses to wild type and BA.5 were also elicited. ELISpot assay showed robust cellular immune response, which was not obviously affected by virus variation. In conclusion, SYS6006 heterologous boosting demonstrated long-term good safety and immunogenicity in participants who had received two or three doses of SARS-CoV-2 inactivated vaccine.
Subject(s)
COVID-19 Vaccines , COVID-19 , Immunogenicity, Vaccine , Humans , Antibodies, Neutralizing , Antibodies, Viral , China , COVID-19/prevention & control , Immunoglobulin G , mRNA Vaccines , Vaccines, InactivatedABSTRACT
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) mRNA vaccine enables quick upgrade of antigen sequence to combat emerging new variants. In an observer-blinded, randomized, placebo-controlled phase 2 trial, immunologically naïve 300 adults and 150 older participants were enrolled and randomized (1:1:1) to receive two doses of 20 µg or 30 µg of a SARS-CoV-2 mRNA vaccine (SYS6006) or placebo. Adverse events (AEs) were recorded through 30 days after the second dose. Live virus neutralizing antibody (Nab), S1 protein-specific binding antibody (S1-IgG) and cellular immunity were tested. Results showed that robust wild-type Nab response was elicited with geometric mean titers of 91.3 and 84.9 in the adults, and 74.0 and 115.9 in the elders, 14 days following the second dose (Day 35) in the 20-µg and 30-µg groups, respectively. All seroconverted for wild-type Nab except two participants. Nab against Omicron BA.5 was mild. Robust wild-type S1-IgG response was induced with geometric mean concentrations of 2751.0 and 3142.2 BAU/mL in adults, and 2474.1 and 2993.5 BAU/mL in elders at Day 35 in the 20-µg and 30-µg groups, respectively. S1-IgG against Omicron BA.2 was induced. Cellular immunity was elicited, particularly in enzyme-linked immunospot assay. The most frequent AEs were injection-site pain and fever. Most reported AEs were grade 1 or grade 2. The AE incidences were similar following the first dose and second dose. No vaccination-associated serious AE was reported. In conclusion, two-dose vaccination with SYS6006 demonstrated good safety, tolerability and immunogenicity in immunologically naïve healthy participants aged 18 years or more.
Subject(s)
COVID-19 Vaccines , COVID-19 , Adult , Aged , Humans , Antibodies, Neutralizing , Antibodies, Viral , China , COVID-19/prevention & control , COVID-19 Vaccines/adverse effects , Double-Blind Method , Healthy Volunteers , Immunogenicity, Vaccine , Immunoglobulin G , mRNA Vaccines , SARS-CoV-2ABSTRACT
The immunity potency upon natural infection or vaccination is the main concern for the vaccine strategy of severe acute respiratory syndrome coronavirus 2 (SARS COV-2 variant), especially the recently reported Omicron variant (B.1.1.529). In this study, 200 recipients immunized with three doses of a COVID-19-inactivated vaccine were enrolled, whose serum samples were collected within 2 months after the third immunization. The neutralizing activity of sera against the pseudotyped Omicron variant, prototype, and Delta variant was determined. Our results demonstrated that the positive neutralization activity was 95.5% for the Omicron variant, 99.5% for the prototype, and 98.5% for the Delta variant. The geometric mean titers (GMT) for the Omicron variant was 49 and maintained sustained immune levels for 2 months, which decreased by 4.9-fold and 3.0-fold compared with the prototype (GMT, 239) and Delta variant (GMT, 148), respectively. In summary, our study demonstrated that three doses of a COVID-19-inactivated vaccine effectively yielded potent cross-neutralizing activity against the Omicron variant at 2 months after the third vaccination.
Subject(s)
COVID-19 Vaccines , COVID-19 , Antibodies, Neutralizing , Antibodies, Viral , COVID-19/prevention & control , Humans , SARS-CoV-2/geneticsABSTRACT
Utilizing the inherent high nitrogen content in natural microalgae to produce value-added nitrogen-containing compounds such as fatty amides and fatty nitriles is a promising method. Herein, a method for producing value-added fatty amides and nitriles by liquefaction of natural microalgae from water blooms in n-heptane was developed. The effects of temperature, metal oxide catalyst (ZrO2 , Al2 O3 , TiO2 , ZnO, MgO, CaO), catalyst amount, and reaction time on the preparation of value-added nitrogen-containing compounds were studied. Under the optimized conditions (0.3â g ZrO2 , 300 °C, 6â h), the total yield of fatty amides was 6.9â wt %, and the yield of fatty nitriles was 1.9â wt %. Compared with the results obtained in the absence of ZrO2 , after adding ZrO2 the total yield of fatty acids was reduced by 4.7â wt % (18.5 to 13.8â wt %), while the total yield of fatty amides only increased by 0.9â wt % (6.0 to 6.9â wt %) and fatty nitriles was increased by 1.5â wt % (0.4 to 1.9â wt %). Exploring the role of ZrO2 by using model compounds (i. e., palmitic acid and palmitamide) revealed that ZrO2 could promote the dehydration of fatty amides to form fatty nitriles, but had limited effect on the reaction of fatty acid and NH3 .
Subject(s)
Biological Products/chemistry , Microalgae/chemistry , Nitrogen Compounds/chemistry , Zirconium/chemistry , Ammonium Compounds/chemistry , Catalysis , Fatty Acids/chemistry , Palmitic Acid/chemistry , Palmitic Acids/chemistry , Temperature , WaterABSTRACT
As the first defence for cells to counteract the toxicity of active oxygen, superoxide dismutase (SOD) plays an important role in the response of living organisms to stress and cell differentiation. One extracellular Cu-ZnSOD (ecCu-ZnSOD), and two MnSODs, were identified based on the Volvariella volvacea genome sequence. All three genes have complicated alternative splicing modes during transcription; only when the fourth intron is retained can the Vv_Cu-Znsod1 gene be translated into a protein sequence with SOD functional domains. The expression levels of the three sod genes in the pilei are higher than in the stipe. The Vv_Cu-Znsod1 and the Vv_Mnsod2 are co-expressed in different developmental stages of the fruiting body, with the highest level of expression in the pilei of the egg stage, and they show a significant, positive correlation with the efficiency of karyogamy, indicating the potential role of these two genes during karyogamy. The expression of the ecCu-Znsod and two Vv_Mnsod genes showed a significant up-regulated when treated by cold stress for one hour; however, the lack of the intracellular Cu-ZnSOD encoding gene (icCu-Znsod) and the special locus of the ecCu-Znsod gene initiation codon suggested a possible reason for the autolysis phenomenon of V. volvacea in cold conditions.
Subject(s)
Cold-Shock Response , Fruiting Bodies, Fungal/enzymology , Fungal Proteins/metabolism , Superoxide Dismutase/metabolism , Volvariella/enzymology , Amino Acid Sequence , Fruiting Bodies, Fungal/metabolism , Fungal Proteins/genetics , Gene Expression Regulation, Developmental , Gene Expression Regulation, Fungal , Molecular Sequence Data , Superoxide Dismutase/chemistry , Superoxide Dismutase/genetics , Volvariella/genetics , Volvariella/growth & developmentABSTRACT
Volvariella volvacea, usually harvested in its egg stage, is one of the most popular mushrooms in Asia. The rapid transition from the egg stage to elongation stage, during which the stipe stretches to almost full length leads to the opening of the cap and rupture of the universal veil, and is considered to be one of the main factors that negatively impacts the yield and value of V. volvacea. Stipe elongation is a common phenomenon in mushrooms; however, the mechanisms, genes and regulation involved in stipe elongation are still poorly understood. In order to study the genes related to the stipe elongation, we analyzed the transcription of laccase genes in stipe tissue of V. volvacea, as some laccases have been suggested to be involved in stipe elongation in Flammulina velutipes. Based on transcription patterns, the expression of Vvlcc3 was found to be the highest among the 11 laccase genes. Moreover, phylogenetic analysis showed that VvLCC3 has a high degree of identity with other basidiomycete laccases. Therefore, we selected and cloned a laccase gene, named Vvlcc3, a cDNA from V. volvacea, and expressed the cDNA in Pichia pastoris. The presence of the laccase signature L1-L4 on the deduced protein sequence indicates that the gene encodes a laccase. Phylogenetic analysis showed that VvLCC3 clusters with Coprinopsis cinerea laccases. The ability to catalyze ABTS (2,2'-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) oxidation proved that the product of the Vvlcc3 gene was a functional laccase. We also found that the expression of the Vvlcc3 gene in V. volvacea increased during button stage to the elongation stage; it reached its peak in the elongation stage, and then decreased in the maturation stage, which was similar to the trend in the expression of Fv-lac3 and Fv-lac5 in F. velutipes stipe tissue. The similar trend in expression level of these laccase genes of F. velutipes suggested that this gene could be involved in stipe elongation in V. volvacea.
Subject(s)
Basidiomycota/genetics , Cloning, Molecular , Gene Expression , Laccase/genetics , Amino Acid Sequence , Basidiomycota/metabolism , Fruiting Bodies, Fungal/genetics , Gene Expression Profiling , Gene Expression Regulation, Fungal , Gene Order , Genetic Loci , Genome, Fungal , Laccase/chemistry , Laccase/metabolism , Molecular Sequence Data , Multigene Family , Phylogeny , Sequence Alignment , TranscriptomeABSTRACT
The formation of fruiting body in Volvariella volvacea is affected by endogenous genes and environmental factors. However, its regulation at a molecular level is still poorly understood. To study the genes involved in the formation of fruiting body, we cloned a new regulator of the G protein signaling (RGS) encoding gene (rgs) from V. volvacea. Phylogenetic analysis showed that RGS in V. volvacea and other basidiomycete RGS proteins from Schizophyllum commune and Coprinus cinereus belong to the same clade. In addition, we assayed intracellular cAMP content in the three developmental stages (mycelium, fruiting body primordia, and button). We also found that the expression of rgs was highly positively correlated to the content of intracellular cAMP during fruiting body formation. The conserved protein sequences and expression of rgs, together with high concent of cAMP at primordia tissue, suggested that rgs gene and cAMP may play a crucial role in fruiting body formation in V. volvacea.
Subject(s)
Cyclic AMP/metabolism , Fruiting Bodies, Fungal/genetics , Fruiting Bodies, Fungal/metabolism , Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Volvariella/genetics , Volvariella/metabolism , Cloning, Molecular , Fungal Proteins/metabolism , Gene Expression Profiling , Gene Order , Phylogeny , Transcriptome , Volvariella/classificationABSTRACT
Fungal laccases play important roles in matrix degradation. Eleven laccase genes, including three novel ones (designated lac1, lac2 and lac4) were identified after sequencing the entire genome of the edible, white-rot fungus Flammulina velutipes. Analysis using bioinformatics revealed that all of the laccases, except lac3, possess a signal peptide. These laccase proteins consist of 502-670 amino acids and have predicted molecular weights ranging from 55kDa to 74kDa. These proteins each contain four copper-binding sites, except for Lac10. Transcriptomes were sequenced at different developmental stages and in different fruiting body tissues to analyze if there was differential expression of laccase genes. The novel laccase gene lac4 exhibited the highest expression levels among all of the observed laccases at every developmental stage and in all fruiting body tissues examined. We conclude that laccases in F. velutipes play a role not only in lignin degradation, but also in fruiting body formation and development.
Subject(s)
Basidiomycota/genetics , Flammulina/genetics , Fungi/genetics , Multigene Family , Amino Acid Sequence , Cloning, Molecular , Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Laccase/genetics , Molecular Sequence Data , Sequence Alignment/methods , TranscriptomeABSTRACT
OBJECTIVE: Based on the analysis of omics data of Volvariella volvacea, a gene encoding glutathione S- transferase (GSTs) named vv-gtol was obtained. To reveal the role of GSTs in the growth and development in edible fungi, the structure, the sequence characters and the expression profile of a GST gene vv-gto1 of Volvariella volvacea were analyzed. METHODS: ZOOM software was used to map sequencing read (reads) from genome and transcriptome against the splicing sequence of genome, to confirm the complete length and the accuracy of the gene sequence, and to visualize gene structure. The MEGA 5.1 was used to do the multiple sequence alignment and phylogenetic tree analysis. Real time fluorescent quantitative PCR was used to determine the expression levels of vv-gtol at different growth periods of Volvariella volvacea. RESULTS: The full sequence of vv-gtol covered 2083 bp, containing 11 exons and 10 introns, and encoded a protein with 356 amino acids. 5'UTR was 305 bp which contains one intron region, and 3'UTR was 86bp. Two intron retentions could be recognized during RNA processing, and the transcripts formed by the intron retention could not translate the correct conservative functional domains. The full-length of vv-gtol had more than 50 accurate positioning genome sequencing reads, suggesting that genome sequencing and assembly results are accurate and reliable. The phylogenetic tree showed that GTO1 of Volvariella volvacea belonged to the subclass I of the Omega class of glutathione S-transferase superfamily, and had the closest relationship with GTO1 and GTO2 in Phanerochaete chrysosporium. The analysis of digital gene expression profiling, fluorescence quantitative PCR and proteomics showed that vv-gtol had the highest expression level in the heterokaryotic hyphae. CONCLUSIONS: This is the first time to obtain a gene encoding glutathione S-transferase from Volvariella volvacea which belongs to Omega class. Our study showed that the gene may play an important role during the special biological functions of heterokaryotic hyphae. This study also suggested that Volvariella volvacea heterokaryotic hyphae in H1521 had stronger resistance ability than other samples. In addition, vv-gto1 could form different alternative splicesome to regulate gene transcription and translation, and ultimately affect the function of the protein.