Phosphotungstic salt used as an efficient catalyst to rapidly remove RhB from water solution

<p>Ag-modified polyoxometalate [Na(H₂O)]₂[Ag(bpf)]₃[PW₁₂O₄₀] (<b>1</b>) (bpf = 2,4-di(4-pyridyl)furan) was synthesized through a hydrothermal reaction. Single-crystal X-ray diffraction analysis revealed that <b>1</b> consists of a hybrid 3D MOF constructed from Keggin-type polyanionic [PW₁₂O₄₀]^3- and cationic Ag-bpf units. Ligand bpf in <b>1</b> was <i>in situ</i> transferred from 1,3-bis(4-pyridyl)propane under the higher temperature and pressure of hydrothermal conditions. Catalytic performance of <b>1</b> as a heterogeneous catalyst for degradation of organic dye Rhodamine B (RhB) was investigated. Experimental results showed that hybrid <b>1</b> was especially active to catalytically degrade RhB under room temperature and natural light. RhB-containing solution (5.0 mg·L⁻¹) could be quickly bleached in a short time, and the high removal efficiency (97%) could be reached in a mere 30 min. The degradation mechanism of RhB was discussed.</p

Image_1_Hemagglutinin expressed by yeast reshapes immune microenvironment and gut microbiota to trigger diverse anti-infection response in infected birds.tif

IntroductionThe H5N8 influenza virus is a highly pathogenic pathogen for poultry and human. Vaccination is the most effective method to control the spread of the virus right now. The traditional inactivated vaccine, though well developed and used widely, is laborious during application and more interests are stimulated in developing alternative approaches.MethodsIn this study, we developed three hemagglutinin (HA) gene-based yeast vaccine. In order to explore the protective efficacy of the vaccines, the gene expression level in the bursa of Fabricius and the structure of intestinal microflora in immunized animals were analyzed by RNA seq and 16SrRNA sequencing, and the regulatory mechanism of yeast vaccine was evaluated.ResultsAll of these vaccines elicited the humoral immunity, inhibited viral load in the chicken tissues, and provided partial protective efficacy due to the high dose of the H5N8 virus. Molecular mechanism studies suggested that, compared to the traditional inactivated vaccine, our engineered yeast vaccine reshaped the immune cell microenvironment in bursa of Fabricius to promote the defense and immune responses. Analysis of gut microbiota further suggested that oral administration of engineered ST1814G/H5HA yeast vaccine increased the diversity of gut microbiota and the increasement of Reuteri and Muciniphila might benefit the recovery from influenza virus infection. These results provide strong evidence for further clinical use of these engineered yeast vaccine in poultry. </p

List of defects on intron splicing with the deletion of <i>MoSRP1</i>.

(XLSX)</p

MoSrp1 enhances the intron splicing efficiency of a virulence gene <i>MoATF1</i>.

(A) Position of the 5’-GUAG-3’ motif in the MoATF1 pre-mRNA. (B) RT-PCR analysis showing splicing efficiency of the first intron of MoATF1 in the ΔMosrp1 mutant srp1ko1 as compared with the wild-type P131. The ratio between the spliced and the total introns was shown at the bottom of each line. (C) RNA-EMSA assay showing the binding strength between MoSrp1 and the 5’-GUAG-3’ consensus in the first intron of MoATF1. (D) RT-PCR analysis showing splicing efficiency of MoATF1 or MoATF1CAAC in strains wild-type (WT) Guy11, ΔMoatf1, MoATF1/ΔMoatf1, and MoATF1CAAC/ΔMoatf1 (mutation of 5’-GUAG-3’ to 5’-CAAC-3’ in MoATF1 transcript). Guy11 was used as the wild-type strain to generate the ΔMoatf1 mutant. The ratio between the spliced and the total introns was shown at the bottom of each line. (E) Colony formed by strains WT, ΔMoatf1, MoATF1/ΔMoatf1, and MoATF1CAAC/ΔMoatf1 on OTA plates at 5 dpi. (F) Statistics on colony diameters of strains WT, ΔMoatf1, MoATF1/ΔMoatf1, and MoATF1CAAC/ΔMoatf1. (G) Conidia per petri dish produced by strains WT, ΔMoatf1, MoATF1/ΔMoatf1, and MoATF1CAAC/ΔMoatf1 on OTA plates. The means and standard deviations in (F and G) were calculated based on three independent experiments each with three plates measured. Asterisk marks a significant difference between the Guy11 and mutant strains using t-test (p (H) Representative disease lesions on leaves of susceptible barley sprayed with conidia of strains WT, ΔMoatf1, MoATF1/ΔMoatf1, and MoATF1CAAC/ΔMoatf1.</p

The RRM domain is essential to all the MoSrp1 functions but the SR region is important for stress responses.

(A) The yeast two-hybrid assay showing that the interactions between N-terminus (1–90 aa) or the C-terminus (91–206 aa) of MoSrp1 and MoRnps1, MoGrp1, and MoThoc1. The interaction between AD-T and BD-Lam is used as the negative control, and the interaction between AD-T and BD-53 is used as the positive control. (B) The colonies formed on OTA plates at 5 dpi by the strains P131, srp1ko1, cSRP1 and srp1ko1 transformants expressing the N-terminal fragments cSRP11-90 (1–90 aa), cSRP11-130 (1–130 aa), the C-terminus fragment cSRP191-206 (91–206 aa), mutated MoSrp1S117A (M117), mutated MoSrp1S119A (M119), and mutated MoSrp1S117AS119A (M117/119) of MoSrp1. (C) Statistics on colony diameters of the strains described in (B). (D) Conidia per petri dish produced on OTA plates by the strains described in (B). (E) Representative disease lesions on the leaves of susceptible barley sprayed with conidia of the strains described in (B), and photographed at 5 dpi. (F) Statistical analysis of colony growth reduction rates of the strains in (B) cultured on CM plates supplemented with different stress agents (1 M sorbitol, 100 μg/ml CFW, or 200 μg/ml CR) at 5 dpi. (G) Statistical analysis of colony growth reduction rates of the strains described in (B) under MM, MM-N, and MM-C conditions. The means and standard deviations in (C, D, F and G) were calculated based on three independent experiments with measuring three plates for each replicate. Asterisk marks significant difference between P131 and mutant strains using t-test (p < 0.05).</p

MoSrp1 is important for vegetative growth, conidia morphology, and conidiation.

(A) Colonies formed by the wild-type (WT) strain P131, the ΔMosrp1 mutant srp1ko1, and its complementation transformant cSRP1 on OTA plates cultured for 5 d. (B) Statistics on colony diameters of strains P131, srp1ko1, and cSRP1 at 5 dpi. The mean and standard deviations were calculated based on three independent experiments each with three plates. Asterisk marks significant difference of the mutant from P131 and cSRP1 using t-test (p (C) Conidia per petri dish produced by strains P131, srp1ko1, and cSRP1. The mean and standard deviations were calculated based on three independent experiments each with three plates. Asterisk marks a significant difference of srp1ko1 from WT and cSRP1 using t-test (p (D) Conidia of strains P131 and srp1ko1 co-stained with CFW and DAPI. At the right of the images were percentages of conidia with 1-, 2-, and 3-cell formed by strains P131 and srp1ko1. At least 300 conidia were calculated for each strain. Bar, 10 μm. (E) Relative expression level of seven cell cycle-related genes in vegetative hyphae of strains P131 and srp1ko1. The gene expression level in P131 was arbitrarily set to 1. The actin gene was used as the internal control. The data were obtained from three replicates. Asterisk marks a significant difference of srp1ko1 from P131 using t-test (p < 0.05).</p

MoSrp1 binds to the GUAG consensus in pre-mRNAs.

(A) Bit image of the top MoSrp1-binding motif by analyzing the RIP-Seq data. (B) RNA-EMSA assay showing the binding of MoSrp1 to the GUAG consensus. Lane 1–4: 2 nM biotin-tagged RNA fragment of 5’-GUAG-3’ with increased amount of MoSrp1 (0.3, 0.6, 0.9, and 1.2 μg); Lane 5–8: 2 nM biotin-tagged RNA fragment and 0.5 μg MoSrp1 with increased amount of unlabeled RNA oligomer (0, 8, 20, and 40 nM); Lane 9: 2 nM biotin-tagged 5’-GUAG-3’ RNA fragment without unlabeled RNA oligomer and MoSrp1. MoSrp1 was expressed and purified from E. coli, and the arrow indicates the binding band. (C) Different types (Type A to N) of aberrantly spliced transcripts with or without the GUAG consensus in the introns and their neighboring exons. A total of 1,066 introns retained in the ΔMosrp1 mutant srp1ko1 were used for analysis. (D) The number of intron retention events with PSI > 0.03 in srp1ko1 in comparison with WT P131. The number of introns with up- and down-regulated PSI was shown in red and yellow, respectively. (E) Distributions of the GUAG consensus within intron (maximum length of the MoSrp1-regulated introns is 330 nt) and its neighboring 5’- and 3’-exon with 100 nt.</p

MoSrp1 regulates intron splicing efficiency.

RT-PCR analyses show (A) enhanced splicing efficiencyand (B) suppressed splicing efficiency of three introns in the wild-type P131, the ΔMosrp1 mutant srp1ko1, and its complementation transformant cSRP1K78R/E80Q expressing the mutated sumoylation site (srp1ko1/Srp1K78R/E80Q-GFP) allele. The corresponding genomic DNA (gDNA) fragment amplified was used as a control with the un-spliced intron. The ratio between the spliced and the total introns was shown at the bottom of each line.</p

List of differential expressed genes with the deletion of <i>MoSRP1</i>.

(XLSX)</p

Expression and purification of MoSrp1 from <i>E</i>. <i>coli</i>.

Lane M, molecular mass markers; Lane–IPTG, total protein extracted before IPTG induction; Lane +IPTG, total protein extracted after IPTG induction; Lane Total Protein, supernatant of total protein extracted after centrifugation; Lane Purified, MoSrp1 protein after affinity chromatography and gel filtration. Lane Anti-His, immunoblotting confirmation of purified MoSrp1 by an anti-His antibody. (TIF)</p