Journal: bioRxiv

Article Title: Simultaneous and sequential multi-species coronavirus vaccination

doi: 10.1101/2022.05.07.491038

Figure Lengend Snippet: (A) Schematics of mRNA vaccine construct design against pathogenic human coronavirus species. Each construct has regulatory elements (5’UTR, 3’UTR and polyA) and spike ORF. The domain structures as well as engineered mutations of translated spike proteins of SARS-CoV-2 Delta variant (Delta), SARS-CoV (SARS) and MERS-CoV (MERS). (B) Engineered mutations in spike protein structures of SARS-CoV-2 Delta, SARS-CoV and MERS-CoV. The N-terminal domain (NTD, blue), receptor binding domain (RBD, green) and S2 subunit (orange) of one protomer along with homologous HexaPro mutations (pink) and Delta variant mutations (red) were highlighted in the spike trimer structures. (C) Schematics of characterization of LNP-mRNA vaccine formulations. Assembly procedure of LNP-mRNA vaccine on NanoAssemblr Ignite and downstream biophysical characterization assays. (D) Histogram displaying radius distribution of LNP-mRNA formulations of SARS-CoV-2 Delta and a Triplex (Delta + SARS + MERS) (abbreviated as Triplex-CoV or Triplex), measured by dynamic light scattering (DLS). The polydispersity index and mean radius of each LNP sample were shown at top left corner. (E) Transmission electron microscope (TEM) images of Delta and Triplex-CoV LNP-mRNAs. (F) Surface expression of functional spike proteins in 293T cells after electroporation of corresponding mRNA, as detected by human ACE2 or human DPP4 Fc fusion protein bound to PE anti-Fc antibody. (G) Schematics of vaccination schedule of the Triplex LNP-mRNA formulations, as well as downstream assays to evaluate the antibody responses and other immunological profiles. (H) Binding antibody titers of plasma samples from mice administered with PBS or different LNP-mRNAs (n = 9) against RBD or ectodomain (ECD) of SARS-CoV-2 wild type (WT, Wuhan/WA-1), Delta variant, SARS and MERS spikes. The binding antibody titers were quantified by area under curve of log 10 -transformed titration curve (log 10 AUC) in . The mice were intramuscularly injected with two doses (x2, 2 weeks apart) of PBS, 1μg SARS-CoV-2 Delta variant LNP-mRNA (delta), 1μg or 3μg equal mass mixture of Delta, SARS and MERS LNP-mRNA (Triplex-CoV). Notes: In the dot-box plots of this figure, each dot represents data from one mouse. Data are shown as mean ± s.e.m. plus individual data points in plots. Two-way ANOVA with Tukey’s multiple comparisons test was used to assess statistical significance. Statistical significance labels: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. Non-significant comparisons are not shown, unless otherwise noted as n.s., not significant.

Article Snippet: To detect surface-protein expression, the cells were stained with ACE2–Fc chimera (Genscript, Z03484) or DPP4-Fc (Sino Biological, 10688-H01H) in MACS buffer (D-PBS with 2 mM EDTA and 0.5% BSA) for 30 min on ice.

Techniques: Construct, Variant Assay, Binding Assay, Transmission Assay, Microscopy, Expressing, Functional Assay, Electroporation, Transformation Assay, Titration, Injection

Journal: Viruses

Article Title: Phage-Displayed Mimotopes of SARS-CoV-2 Spike Protein Targeted to Authentic and Alternative Cellular Receptors

doi: 10.3390/v14020384

Figure Lengend Snippet: Characteristics of recombinant human cell receptors and viral spike proteins.

Article Snippet: hDPP4 , Sino Biological , 10688-H08H , D34 , P766 , 744 , 86.3.

Techniques: Recombinant, Molecular Weight

Journal: Pathogens and Disease

Article Title: Protective vascular coagulation in response to bacterial infection of the kidney is regulated by bacterial lipid A and host CD147

doi: 10.1093/femspd/fty087

Figure Lengend Snippet: Epithelial signaling factors induce endothelial TF activation. ( A ) Proteome array of supernatant of epithelial cells infected with CFT073 for 4 h. Supernatant from uninfected cells was used as the comparative control. Bars represent index of mean pixel density from the nitrocellulose membranes, n = 3. ( B ) CD26 and ( C ) CD147 expression in the supernatant of RPTEC/TERT1 cells stimulated for 4 h with cell culture medium, 5 mg/ml LPS or infected at an MOI 1:20 with CFT073 or LT002. Measurements were performed by ELISA, n = 5. ( D ) TF pro-coagulant activity, pM, measured using chromogenic assays in endothelial cells treated with the noted concentrations of ( D ) CD26, or ( E ) CD147, n = 3. All bars represent mean ± SD.

Article Snippet: Endothelial cells were stimulated with homo-dimer human recombinant EMMPRIN (Abcam, Sweden #AB155636) or human CD26/DPP4 recombinant protein (Sino Biological #10 688-HNCH-10) for 4 h before TF activation determination at the noted concentrations.

Techniques: Activation Assay, Infection, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Activity Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin improves left ventricular relaxation in obese ZSF1 rats. Plasma DPP‐4 activity (A), aGIP levels (B), fasting glucose levels (C), bodyweight (D), mitral valve deceleration time (E) and E/E’ ratio (F) in 20‐wk‐old linagliptin‐ (Obese + Lina) and placebo‐treated obese (Obese) ZSF1 rats (n = 7 per group). aGIP, active glucose‐dependent insulinotropic peptide; DPP‐4, dipeptidyl peptidase‐4; E, early mitral inflow peak velocity; E’, early diastolic mitral annulus peak velocity; ND, not detected; RFU, relative fluorescence units. Data are expressed as mean ± SEM. Data were analysed using a two‐tailed unpaired Student t test, except D was analysed by a Mann‐Whitney U test. * Indicates P < .05, ** P < .01 and *** P < .001

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: Activity Assay, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin decreases F passsive by increasing titin phosphorylation at N2Bus S4010 in human cardiomyocytes in vitro. A, Cardiomyocyte passive stiffness in human cardiomyocytes treated with 300 ng/mL DPP‐4, 100 nmol/L linagliptin and 300 ng/mL DPP‐4, or control (DMSO/PBS) for 30 min (n = 3 different left ventricular tissues measuring at least 12 cardiomyocytes from each left ventricular tissue per condition). B, PKA‐mediated N2Bus S4010 phosphorylation and representative Coomassie Blue stained‐PVDF membranes and (C) PKG‐dependent N2Bus S4099 phosphorylation in human cardiomyocytes (n = 5 different left ventricular tissues) treated with vehicle control (DMSO/PBS), 300 ng/mL DPP‐4, 300 ng/mL DPP‐4 and 100 nmol/L linagliptin, or 100 nmol/L linagliptin for 2 h. F passive , passive stiffness; N2Bus, N2B unique sequence. Panel A was analysed using a two‐tailed unpaired Student t test with * P < .05. Panel B and C were analysed using a one‐way ANOVA with Dunnett's multiple comparison post hoc test with * P < .05 and *** P < .001 comparing control, DPP‐4 + Lina and Lina to DPP‐4

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: In Vitro, Staining, Sequencing, Two Tailed Test

Journal: Journal of Cellular and Molecular Medicine

Article Title: Linagliptin prevents left ventricular stiffening by reducing titin cleavage and hypophosphorylation

doi: 10.1111/jcmm.16122

Figure Lengend Snippet: Linagliptin prevents DPP‐4‐mediated titin cleavage in human cardiomyocytes in vitro. A‐B, Western blots of total titin, containing the two isoforms N2BA and N2B and the known degradation product titin‐2 (T2) of human intact non‐permeabilized (A) and non‐intact permeabilized (B) human cardiomyocytes exposed to control PBS/DMSO (Ctrl) for 0 or 2 h (lane 1 and 2, respectively), 300 ng/mL DPP‐4 for 30 min and 2 h (lane 3 and 4, respectively), 300 ng/mL DPP‐4 and 100 nmol/L linagliptin for 2 h (lane 5) or 100 nmol/L linagliptin alone for 2 h (lane 6) in vitro (n = 3 different left ventricular tissues). Titin cleavage is indicated by red asterisks. Western blots of the specific N2Bus region (C; cross‐species conserved sequence QELLSKETLFP) and PEVK region (D; cross‐species conserved sequence KLRPGSGGEKPP) of human cardiomyocytes exposed to control PBS/DMSO (Ctrl) for 0 or 2 h (lane 1 and 2, respectively), 300 ng/mL DPP‐4 for 30 min and 2 h (lane 3 and 4, respectively), 300 ng/mL DPP‐4 and 100 nmol/L linagliptin for 2 h (lane 5) or 100 nmol/L linagliptin alone for 2 h (lane 6) (n = 3 different left ventricular tissues). Titin cleavage is indicated by red asterisks. The framed area represents the location for the zoomed images presented on the right side. Ctrl, control; DPP‐4, dipeptidyl peptidase‐4; F passive , passive stiffness; Lina, linagliptin; N2Bus, N2B unique sequence. Data are expressed as mean ± SEM

Article Snippet: Stripped tissue was incubated with (a) 300 ng/mL recombinant his‐tagged DPP‐4 (# 50718‐M08H; Sino Biological), (b) 300 ng/mL recombinant his‐tagged DPP‐4 and 100 nmol/L linagliptin (Boehringer Ingelheim GmbH) , or (c) PBS/DMSO (control) for 30 minutes to assess F passive in vitro.

Techniques: In Vitro, Western Blot, Sequencing