Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Serum ACE2 activity is significantly correlated with SBP in stroke-alert patients and healthy young adults, but not AIS patients. Correlation graphs of ACE2 activity and SBP among stroke-alert patients (a) and healthy young adults (b) as compared to stroke patients (c). Young adult blood plasma samples in panel (b) were from a biorepository established by Wegman et al., which were obtained from research participants undergoing baseline measurements. (d) Correlation graph of ACE activity and mRS at discharge from hospital among AIS patients. ACE2: angiotensin converting enzyme 2; AIS: acute ischemic stroke; mRS: modified Rankin score; RFU: relative fluorescence unit; SBP: systolic blood pressure.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay, Clinical Proteomics, Modification, Fluorescence

Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Activity of ACE2 and ACE in serum is altered following stroke. For human serum, bar graphs are means ± SEM and represent enzyme activity levels of ACE2 (a) and ACE (c) from control, stroke-alert, or AIS patients at an average of 3.6 hours and again at 3 days after stroke. Individual differences and means ± SEM in ACE2 (b) and ACE (d) are shown. * P <0.05 versus control and † P <0.05 versus stroke-alert. ‡ P <0.05 versus AIS <6 hours. ACE: angiotensin converting enzyme; ACE2: angiotensin converting enzyme 2; AIS: acute ischemic stroke; RFU: relative fluorescence unit.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay, Control, Fluorescence

Journal: Journal of the Renin-Angiotensin-Aldosterone System: JRAAS

Article Title: Serum activity of angiotensin converting enzyme 2 is decreased in patients with acute ischemic stroke

doi: 10.1177/1470320316661060

Figure Lengend Snippet: Predictors of acute ischemic stroke by multiple linear regression analysis.

Article Snippet: Reaction Km and Vmax were determined using control samples and recombinant human ACE2 (R&D Systems, Inc., #933-ZN-010) as a positive control, and all samples were run in duplicate.

Techniques: Activity Assay

Journal: Viruses

Article Title: Design of Three Residues Peptides against SARS-CoV-2 Infection

doi: 10.3390/v14102103

Figure Lengend Snippet: BLI interaction analysis. RBD (0.2 µM) ( A , B ) either alone in solution or mixed with VFI ( A ) and TLH ( B ) peptides, tested at different concentrations (40 µM, blue lines; 80 µM, green lines and 160 μM, orange lines). BLI interaction was performed at 25 °C in PBST (10 mM phosphate, 150 mM NaCl, 0.05% Tween 20, pH 7.4). The corresponding plots ( C , D ) of steady-state binding from the end of the association phases (nm), after the subtraction of RBD signal, against analyte concentration were used to calculate the steady-state affinity by nonlinear regression analysis using GraphPad 5 software.

Article Snippet: An Octet ® Red 96 system ® (ForteBio, Fremont, CA, USA) was used to detect interactions between the His-tagged Spike RBD protein (Elabscience Biotechnology, Biomedical Park, Wuhan, China; Cat. No.: PKSH032068) and peptides using Octet His2 biosensors according to the standard instructions with minor modifications.

Techniques: Binding Assay, Concentration Assay, Software

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Schematic illustration of SARS-CoV-2 spike protein and ACE2 receptor binding. (a) SARS-CoV-2 binding to the ACE2 receptor on the host cell surface. (b) Binding of ACE2 and spike protein along with illustration of the spike protein subunits, S1 and S2. (c) Schematic showing the distribution of the spike protein in solution. (d) Schematic showing the distribution of ACE2 in suspension. (e) Distribution of ACE2 and S protein after binding in solution.

Article Snippet: The human ACE2 recombinant protein (ProSci Cat No. 10-114) was resuspended in sterile deionized water at a stock concentration of 500 ng/µl as per the manufacturers instruction and used for the analysis.

Techniques: Binding Assay

Journal: medRxiv

Article Title: Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

doi: 10.1101/2020.11.24.20237628

Figure Lengend Snippet: Optical detection of binding interactions between ACE2 and other proteins. (a) Measured light intensities over time for individual assessment of ACE2, S1X, S1Y, and BSA. (b) The measured mixed light intensities versus time for ACE2 mixed with either S1X, or S1Y, or BSA. (c) The measured ACE2-S1X interaction profile for an extended time period.

Article Snippet: The human ACE2 recombinant protein (ProSci Cat No. 10-114) was resuspended in sterile deionized water at a stock concentration of 500 ng/µl as per the manufacturers instruction and used for the analysis.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Molecular Interactions of Tannic Acid with Proteins Associated with SARS-CoV-2 Infectivity

doi: 10.3390/ijms23052643

Figure Lengend Snippet: Inhibitory effects of different polyphenols on the interaction between SARS-CoV-2 spike protein receptor binding domain (RBD (N501Y)) and human angiotensin-converting enzyme 2 (ACE2). ( A ) 10 µM of pelargonidin-3-O-glucoside (Pel-3-O-G), malvidin-3-O-glucoside (Mal-3-O-G), cyanidin-3-O-glucoside (Cya-3-O-G), peonidin, tannic acid (TA), 1,3,6-tri-O-galloyl-β-D-glucose (TGG), and corilagin were tested to evaluate their ability to inhibit the binding of immobilized spike protein (0.5 µg/mL) to human, biotin-labeled ACE2 (0.25 µg/mL) by using an enzyme-linked immunosorbent assay (ELISA). Dose effect inhibition of 0.1, 1, and 5 µM ( B ) TA, ( C ) TGG, and ( D ) corilagin. The absorbance of ACE2 (0.25 µg/mL) at 450 nm was set to 100%. Results are expressed as mean ± SD (n = 3). Statistical analysis was performed using one-way ANOVA followed by the Tukey post hoc test with * p < 0.05, ** p < 0.01, *** p < 0.001 compared to ACE2 (0.25 µg/mL).

Article Snippet: Biotinylated recombinant human ACE2 with purity >95% and camostat mesylate compound were both purchased from R&D Systems (Minneapolis, MN, USA).

Techniques: Binding Assay, Labeling, Enzyme-linked Immunosorbent Assay, Inhibition

Journal: bioRxiv

Article Title: Anti-CoVid19 plasmid DNA vaccine induces a potent immune response in rodents by Pyro-drive Jet Injector intradermal inoculation

doi: 10.1101/2021.01.13.426436

Figure Lengend Snippet: A cellular immune response and neutralizing antibodies. A) IFN-γ and IL-4 ELISpot responses in splenocytes from rats vaccinated with 240 μg pVAX1-SARS-CoV2-co, with or without re-stimulation with recombinant S1+S2. The p-value calculated using Welch’s t-test between IFN-γ secreting cell number in the antigen-stimulated group and the un-stimulated group was 0.0059 (n=6). B) ACE2 (and S1+S2) binding inhibition assay using 2-to 12-week immunized rat serum normalized to pre-serum, at reciprocal 10-to 31250-fold dilutions. C) Neutralization titers (ID75) of 2-to 12-week immunized rat serum indicated as the dilution of serum required for 75% inhibition of ACE2-S1+S2 binding as shown in (B). D) Inhibition rate against the pseudovirus. The immunized rat serum at 8 weeks was used at reciprocal 20-to 12500-fold dilutions. E) Neutralization activity (ID75; 75% inhibition). The values indicate the inhibitory dose of the serum which shows 75% inhibition rate against pseudovirus binding as shown in (D).

Article Snippet: To analyze the binding inhibition of S1+S2 and ACE2 by neutralizing antibodies in the immunized rat and mouse serum, 96-well plates were coated with human ACE2 recombinant protein (1μg/ml, mFc tag; #83986, Cell Signaling Technology), and then blocked using PBS containing 5% skim milk for 2 h at room temperature.

Techniques: Enzyme-linked Immunospot, Recombinant, Binding Assay, Inhibition, Neutralization, Activity Assay

Journal: bioRxiv

Article Title: Anti-CoVid19 plasmid DNA vaccine induces a potent immune response in rodents by Pyro-drive Jet Injector intradermal inoculation

doi: 10.1101/2021.01.13.426436

Figure Lengend Snippet: Virus challenge on pVAX1-SARS-CoV2-co immunized mice A) Pre-viral challenge of mice immunized with 160 μg of pVAX1-SARS-CoV2-co plasmid intradermally (ID; n = 8) or no-vaccinated (NV: n=10) two times at 2-week intervals. Antibody titer (half maximum) for recombinant S1+S2 in the blood serum 16 weeks post initial vaccination, assessed using ELISA. P <0.001 Student’s t -test. B) Antibody titer (half maximum) for recombinant RBD in the blood serum in ID and NV mice. P =0.018 Student’s t -test. C) Neutralization titer (ID75) for recombinant ACE2-S1+S2 binding inhibition in the blood serum of ID mice. D) ID and NV (n=10) mice were intranasally infected with mouse-adapted SARS-CoV-2 virus. 50% tissue culture infective doses (TCID50) values in the lung tissues of each animal are shown. P =0.016 Student’s t -test. All individual values are shown in .

Article Snippet: To analyze the binding inhibition of S1+S2 and ACE2 by neutralizing antibodies in the immunized rat and mouse serum, 96-well plates were coated with human ACE2 recombinant protein (1μg/ml, mFc tag; #83986, Cell Signaling Technology), and then blocked using PBS containing 5% skim milk for 2 h at room temperature.

Techniques: Virus, Plasmid Preparation, Recombinant, Enzyme-linked Immunosorbent Assay, Neutralization, Binding Assay, Inhibition, Infection

Journal: bioRxiv

Article Title: Anti-CoVid19 plasmid DNA vaccine induces a potent immune response in rodents by Pyro-drive Jet Injector intradermal inoculation

doi: 10.1101/2021.01.13.426436

Figure Lengend Snippet: Individual values of pre-viral challenge antibody titers of ID and NV mice for recombinant S1+S2 (A), RBD (B) and neutralization titer (inhibitory dose at 50% neutralization; ID50) of ACE2-S1+S2 binding inhibition (C), as well as TCID50 values in the mouse lung tissues after virus challenge as described in

Article Snippet: To analyze the binding inhibition of S1+S2 and ACE2 by neutralizing antibodies in the immunized rat and mouse serum, 96-well plates were coated with human ACE2 recombinant protein (1μg/ml, mFc tag; #83986, Cell Signaling Technology), and then blocked using PBS containing 5% skim milk for 2 h at room temperature.

Techniques: Recombinant, Neutralization, Binding Assay, Inhibition, Virus

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: a Preparation of rhACE2-loaded electrospun nanofiber patch by hyaluronan (HA) micro-sol electrospun and the mechanism of formation of core-shell structure. b Myocardial infarction mice model established by precise ligation of the left anterior descending (LAD) coronary artery along with the illustration of rhACE2 patch implantation. c In situ rhACE2 patch niche degrading angiotensin II (AngII) into a cardioprotective heptapeptide, Ang1–7, which counter the AngII/AT1R mediated effects by inhibiting cardiac fibrosis and cardiomyocyte apoptosis. PLLA poly(L-lactic acid), rhACE2 recombinant human angiotensin-converting enzyme 2, AT1R angiotensin II receptor type 1, PKC protein kinase C, MAPK mitogen-activated protein kinase, ECM extracellular matrix.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques: Ligation, In Situ, Recombinant

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: The general view, SEM and TEM micrographs demonstrating the core-shell structure ( a ), dynamic light scattering ( b ), water contact angle ( c ) and stress-strain curve ( d ) from PLLA-HA/ACE2 electrospun nanofibers. Scale bars represent 1 cm in general view, 20 μm in SEM and 1 μm in TEM.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques:

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: a Representative live/dead fluorescence stained by Calcein AM (green) and Propidium Iodide (red) at day 3. Scale bars represented 50 and 20 μm for zoomed pictures. b Live and dead cell count per HPF of control, PLLA, PLLA-HA/ACE2 groups. n = 4/group. c CCK-8 cell viability quantification results in different groups after 1, 2, or 3 days. n = 4/group. Data were represented as the mean ± SEM and analyzed for statistical significance using One-way ANOVA followed by Tukey’s multiple comparison test; NS no significant difference.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques: Fluorescence, Staining, Cell Counting, Control, CCK-8 Assay, Comparison

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: a In vitro releasing curve of PLLA-HA/ACE2 fibrous membranes. b Releasing buffer collected at specified time points during release study was added into culture media of NRCMs undergone 6 h hypoxia. Representative immunofluorescence image of TUNEL (red) and nuclear visualized by DAPI (blue). Scale bars represented 50 μm. c Statistical analysis of TUNEL-positive cell counts. n = 3/group. Data were represented as the mean ± SEM and analyzed for statistical significance using One-way ANOVA followed by Tukey’s multiple comparison test; NS no significant difference; ** P < 0.01 compared to the control group.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques: In Vitro, Immunofluorescence, TUNEL Assay, Comparison, Control

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: a The animal research timeline design with images represented the acute MI model and successful implantation of rhACE2 patch. Echo, echocardiography. b Representative M-mode parasternal long axis view of left ventricle echocardiographic images of different groups at day 28 after LAD coronary artery ligation. Statistical analysis of left ventricular ejection fraction ( c ), shortening fraction ( d ), heart weight/body weight ratio ( e ), LV end-diastolic diameter ( f ), LV end-systolic diameter ( g ) and heart weight/tibial length ( h ) determined by echocardiography obtained from PLLA, intramyocardial injection (IM) and PLLA-HA/ACE2 treatment group at day 7, day 14 and day 28 after operation. n = 5/group. Data were represented as the mean ± SEM and analyzed for statistical significance using two-way ANOVA followed by Tukey’s multiple comparison test; NS no significant difference, * P < 0.05, ** P < 0.01.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques: Ligation, Injection, Comparison

Journal: NPJ Regenerative Medicine

Article Title: Myocardial fibrosis reversion via rhACE2-electrospun fibrous patch for ventricular remodeling prevention

doi: 10.1038/s41536-021-00154-y

Figure Lengend Snippet: H&E stained section ( a ), Masson’s trichrome staining ( b ) and Sirius Red staining ( c ) of mice hearts at low magnification after heart was obtained 28 days after MI. Scale bars represented 500 μm. d Brain natriuretic peptide (BNP) immunohistochemical staining of hearts from PLLA, intramyocardial injection (IM) and PLLA-HA/ACE2 treatment group 28 days after MI. Scale bars represented 40 μm. Morphometric parameters including the percentage of infarct size percentage of total LV area ( e ) and the percentage of infarcted thickness of LV anterior wall ( f ) were measured from the Sirius Red stained slides via ImageJ software. n = 5/group. g The integrated optical density (IOD)/area ratios of BNP were quantified from the immunohistochemical stain by Image-Pro Plus software. n = 5/group. Data were represented as the mean ± SEM and was analyzed for statistical significance using one-way ANOVA followed by Tukey’s multiple comparison test; NS no significant difference, * P < 0.05, **** P < 0.0001.

Article Snippet: Human ACE2 (18-652) recombinant protein (rhACE2, #85054) was purchased from Cell Signaling Technology (USA). rhACE2 contains necessary enzymatic activity associated protein structure and its effectiveness in mice and rats had been previously tested , so it was selected for both our cell and animal studies.

Techniques: Staining, Immunohistochemical staining, Injection, Software, Comparison