Supplementary document for Tuning the persistent luminescence property of Y3Al2Ga3O12:Ce3+,Yb3+ phosphor by controlling the intrinsic oxygen vacancy concentration - 6056742.pdf

PLE and PL spectra, PersL decay curves, and TL spectra of the YAGG:Ce annealed in air and H2/Ar; PersL spectra of the YAGG:Ce,Yb annealed in air and H2/Ar measured at 1, 5, and 10 min after excitation; Comparison of the normalized PersL spectra and

Thermodynamic Allosteric Switch-Actuated 3D DNA Nanomachine for Ultrasensitive Electrochemical/Fluorescent Dual-Mode Biosensing of a Transcription Factor

Herein, we reported an innovative thermodynamic allosteric switch-actuated 3D DNA nanomachine for selective, sensitive, and accurate electrochemical (EC)/fluorescent (FL) dual-mode biosensing of a microphthalmia-associated transcription factor (MITF). The thermodynamic allosteric switch was ingeniously customized as a hairpin probe (HP) that was in dynamic equilibrium but rapidly interconverting conformations. At the “inactive state”, the MITF-binding region and the switch part were “sequestered”. Upon the introduction of MITF, an MITF-HP complex promptly formed, and the equilibrium of HP thermodynamically inclined from the “inactive state” toward the “active state” conformation. Immediately, the exposed switch on HP effectively actuated the 3D DNA nanomachine and synchronously produced the restriction site for Nb.BbvCI nicking endonuclease. After the autonomous conveying of the 3D DNA nanomachine by means of the high-efficiency circularly nicking endonuclease signal amplification (NESA), not only was MB-S1 in the supernatant used for FL measurements but also MB-SP/MNs/S2 in the precipitate was adapted for EC analysis, significantly improving the utilization of output products derived from the 3D DNA nanomachine. Accordingly, benefiting from the efficient DNA nanomachine signal amplification manner and the self-calibration function of a dual-mode bioassay, the constructed biosensor exhibits superior sensitivity and accuracy for MITF determination

Effect of Oxygen Vacancies on the Persistent Luminescence of Y₃Al₂Ga₃O₁₂:Ce³⁺,Yb³⁺ Phosphors

The effect of oxygen vacancies (VO) and the atmosphere influence on persistent luminescence (PersL) in Y3Al2Ga3O12 (YAGG):Ce3+,Yb3+ are investigated by heating it in CO2, air, and 10% H2/90% Ar atmospheres. The VO-rich YAGG phosphors with outstanding PersL are successfully obtained by the common contribution of the reducing atmosphere and the incorporated Yb3+ ions, and the concentration of oxygen vacancies in the phosphors is characterized by X-ray photoelectron spectroscopy and electron paramagnetic resonance measurements. Compared to the best sample prepared in neutral CO2, the reduced sample shows an increase of 30% in initial intensity and 100% in duration time, while the oxidized sample decreases drastically and shows a faint and undetectable PersL. The enhancement is mainly caused by the abundant formation of VO, which is achieved by the pairing of VO with Yb2+ ions. The newly created VO by the reducing calcination is inferred to be adjacent to the Yb site and forms a compensation-type defect cluster due to the charge compensation effect. These findings reveal that understanding the effect and formation of VO is of great significance to design a high-performance phosphor

Controllable Fabrication of Poly(Arylene Ether Nitrile) Dielectrics for Thermal-Resistant Film Capacitors

High-temperature-resistant dielectric films, the heart of energy storage components in film capacitors, are key elements to ensure that the capacitors operate properly in harsh environments. Herein, a kind of flexible thermal-resistant poly­(arylene ether nitrile) (TR-PEN) dielectric film with controllable high-temperature resistance is fabricated through post self-crosslinking of PEN at different temperatures for different times. The dielectric films can present extremely high thermal stability (Tg > 370 °C), a long service life at 300 °C (4.5 × 104 min, 95 wt % of the residual weight), and a low temperature coefficient of dielectric constant from 50 to 300 °C (–4 °C–1). The retention of the energy density of TR-PEN350 at 300 °C is higher than 80% compared with that at room temperature. These results indicate that the TR-PEN films can be used over a long term as capacitor films at temperatures up to 300 °C. Besides, the simple and controllable fabrication technique for the TR-PEN can be easily industrialized

Characterizing the dynamics of BCR repertoire from repeated influenza vaccination

Yearly epidemics of seasonal influenza cause an enormous disease burden around the globe. An understanding of the rules behind the immune response with repeated vaccination still presents a significant challenge, which would be helpful for optimizing the vaccination strategy. In this study, 34 healthy volunteers with 16 vaccinated were recruited, and the dynamics of the BCR repertoire for consecutive vaccinations in two seasons were tracked. In terms of diversity, length, network, V and J gene segments usage, somatic hypermutation (SHM) rate and isotype, it was found that the overall changes were stronger in the acute phase of the first vaccination than the second vaccination. However, the V gene segments of IGHV4-39, IGHV3-9, IGHV3-7 and IGHV1-69 were amplified in the acute phase of the first vaccination, with IGHV3-7 dominant. On the other hand, for the second vaccination, the changes were dominated by IGHV1-69, with potential for coding broad neutralizing antibody. Additional analysis indicates that the application of V gene segment for IGHV3-7 in the acute phase of the first vaccination was due to the elevated usage of isotypes IgM and IgG3. While for IGHV1-69 in the second vaccination, it was contributed by isotypes IgG1 and IgG2. Finally, 41 public BCR clusters were identified in the vaccine group, with both IGHV3-7 and IGHV1-69 were involved and representative complementarity determining region 3 (CDR3) motifs were characterized. This study provides insights into the immune response dynamics following repeated influenza vaccination in humans and can inform universal vaccine design and vaccine strategies in the future.</p

Genomic description of AA 813 in Spike of <i>Sarbecovirus</i>.

(A) The phylogenetic tree based on Sarbecovirus S proteins (SARS-like strains, n = 24 genomes; SARS-CoV strains, n = 13 genomes; SARS-CoV-2 strains, n = 6 genomes). All strains invariantly containing serine at position 813 were marked red while containing threonine were marked mazarine. Color coding as indicated according to species. (B) Amino acid frequency of site 813. 114 complete spike protein sequences of SARS-CoV were collected from NCBI and 10,060,583 complete spike protein sequences of SARS-CoV-2 (2020–2022) were collected from GISAID. The method of analysis was performed as previously described [74]. Briefly, After removing redundant sequences with 100% sequence identity and multiple sequence alignment (MSA), site 813 based on the reference sequence of SARS-CoV-2 was derived and the amino acid frequency of site 813 can be calculated based on the un-redundant dataset of SARS-CoV and SARS-CoV-2, respectively. (C) Phylogenetic tree of Human coronavirus. The representative strains of 7 human coronaviruses were clustered by amino acid sequence phylogeny and observed the diversity of AA 813. In α-CoV (229E and NL63), there was only Alanine, while in β-CoV, Serine was in SARS-CoV-2, MERS-CoV, OC43-CoV and HKU1-CoV; threonine only in SARS-CoV. We used the WAG+F+I+G4 optimal model of the Iqtree software to construct a phylogenetic tree based on S proteins. The right-hand sequence mapping was based on texshade software for mapping. The secondary structure, i.e. the membrane fusion region, was predicted using the PSIPRED web page. (D) Schematic summary. By modifying the usage of TMPRSS2, Spike residue 813 affects the fusogenicity of Sarbecovirus. S813 S has a high utilization and complete the membrane fusion process with a small amount of TMPRSS2, but T813 S has a low ability and needs more TMPRSS2.Although both Serine and Threonine can be found at position 813, the evolutionary trend of Sarbecovirus maybe likely Serine.</p

S813T mutation reduced the use of TMPRSS2 by S protein.

(A) Schematic illustration of SARS-CoV S and SARS-CoV-2 S including proteolytic cleavage sites: S1/S1, S2’ and CTSL cleavage sites in S1. Residue 813 was indicated as red. Arrow heads indicated the cleavage site. (B) Western blot. Top panel: A representative blot of VSVpp digested with TPCK-trypsin (2 μg/ml at 37°C for 30 min), FL-Spike (S) and CL-Spike (S2 and S2’) were marked as indicated. Bottom panel: quantified band intensity using Image J to analyze the protein expression and the ratio of S2 and S2’ to the total S. (C) Spike-based cell-cell fusion assay. With the overexpression of TMPRSS2 (purple) in ACE2-293T, the fusion activity was quantified at different times (2, 6, 12 and 24hpt). The results of S813 S were shown as full lines and T813 S as dotted lines. (D) Representative images of cell-cell fusion. The area of cell fusion was shown as green (up) and black (bottom). Scale bar: 500 μm. (E-G) Fusion inhibition of Camostat and E-64d in ACE2-293T+TMPRSS2. A schematic diagram showing the GFP-split system with the inhibitor Camostat and E-64d for Spike-ACE2/TMPRSS2 mediated cell fusion (E). After being pre-incubated with the indicated concentration (0, 10, 50, 100 μm) of Camostat (F) and E-64d (G), the fusion activity was quantified at different times (2, 6, 12 and 24hpt), and the results of Camostat were shown as purple and E-64d as blue. (H) Spike cleavage assay by TMPRSS2. A representative blot of SARS-CoV-2 (Up panel) or SARS-CoV (Down panel) Spikes digested with TMPRSS2 at 0, 0.1, 0.3, 0.5, 1.0, 2.0 μM in assay buffer, at 37°C for 30 min, FL-Spike (S) and CL-Spike (S2 and S2’) were marked as indicated. Results are means +/- SD from at least three fields per condition. Results are representative of at least three independent experiments. Statistically significant differences (*: p<0.05) between S813 Spike and T813 Spike were determined by Student’s test at each point (C, F and G).</p

The S813T mutation has no effect on S protein interactions with ACE2.

(A) SDS-polyacrylamide gel electrophoresis (PAGE) of SARS2-6P, SARS-2P and Residue 813 substitution S variants. Molecular weight standards are indicated at the left in KD. (B) Competitive ELSIA to detect the binding affinity between S proteins and ACE2. Red line indicated SARS-CoV-2, and blue line indicated SARS-CoV. (C) Neutralization curves for RBD representative antibodies, m396 and X65, with the VSVpp containing parent and mutant S proteins. Each point represents the mean and standard error of 2 independent measurements.</p

The Influence of TMPRSS2 on Spike fusogenicity.

(A) Spike-based fusion assay. The fusion activity was quantified by measuring the ratio of GFP+ area to DAPI area by imaging at different concentration of TMPRSS2 (0, 0.5, 1 and 2 μg). The results for SARS-CoV-2 and SARS-CoV were shown as Red and grey lines, respectively. (B) Representative images of cell-cell fusion. Scale bar: 500 μm. (C) Western blot. A representative blot of 293T cell lysates expressing TMPRSS2 with various concentrations. Beta-actin was used as a control. (D) Cell viability assay. The cell viability with different doses of Camostat and E64d was evaluated by CCK8 assay. Results are means +/- SD from at least three fields per condition. Results are representative of at least three independent experiments. Statistically significant differences (**: p (TIF)</p

Spike S813T mutation of SARS-CoV-2 disturbed fusion in Non-trypsin condition.

The fusion activity of Spikes with S813 or T813 were quantified in non-trypsin condition, by measuring the ratio of GFP+ area to DAPI area at different concentration: Spike expressing cells mixing with 293T cells, ACE2-293T cells or ACE2-293T/TMPRSS2 cells respectively. The results for S813 S and T813 S were shown as Red and Blue histogram, and the disturbance of fusion by S813T mutation is still observed in SARS-CoV-2, but not in SARS-CoV. After overexpressing TMPRSS2, the fusion ability increased both in SARS-CoV and SARS-CoV-2, and the influence of S813T mutation was disappeared. Representative cell-cell fusion images of SARS-CoV-2 (up) and SARS-CoV (down) were shown. Scale bar: 500 μm. Results are means +/- SD from at least three fields per condition. Results are representative of at least three independent experiments. Statistically significant differences (**: p (TIF)</p