biotinylated goat anti human ace 2 antibody cat dy933 05  (R&D Systems)

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: ( A ) Analysis of weight loss (%) and ( B ) clinical score (daily sum) of uninfected control and SARS-CoV-2-infected K18-hACE2 mice treated with Losartan or Losartan + Lisinopril combination. UU = Untreated Uninfected, CTI = Combination-treated Infected, LTI = Losartan-treated Infected, and UI = Untreated Infected. Signs indicate significant differences: (#) all groups versus UU group; (&) UI versus UU group; (*) UI versus CTI or LTI: * p < 0.05, ** p < 0.005, *** p < 0.0005 and **** p < 0.0001. Mice in the UU group showed clinical scores of zero at all times. Data presented as mean ± standard error of the mean (SEM). Analysis performed by mixed-effects model with Geisser–Greenhouse correction, followed by Tukey’s post hoc test. n = 30 per group.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Control, Infection

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: Assessment of lung capacity through the parameters of respiratory frequency ( A ), Rpef ( B ), minute volume (Mv) ( C ), and Pause ( D ) in uninfected control or SARS-CoV-2 infected K18-hACE2 mice treated or not with Losartan or Losartan + Lisinopril combination at 6/7 days post-infection. UU = Untreated Uninfected, CTI = Combination-treated Infected, LTI = Losartan-treated Infected, and UI = Untreated Infected. The normality of the data was assessed using the Shapiro–Wilk test. Data of ( A – C ) showed normal distribution and were analyzed using a parametric test, one-way ANOVA followed by the Bonferroni post-test. Data of ( D ) did not show normal distribution, and a non-parametric analysis was performed, with the Kruskal–Wallis test followed by the Dunn post-test. (#) Significant difference, all groups versus the UU group. (*) Significant difference between specific groups as indicated by the lines: * p < 0.05, ** p < 0.005, *** p < 0.0005 and **** p < 0.0001. n = 10 per group.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Control, Infection

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: ( A ) Quantification of ACE2, on lungs, by ELISA in uninfected mice treated or not with Losartan or Losartan + Lisinopril combination over 21 days (black and white columns; UU: uninfected untreated; CTU: combination-treated, uninfected; LTU: Losartan-treated, uninfected) and in the same animals after infection with SARS-CoV-2 (colored columns; CTI [gray]: combination-treated, infected; LTI [green]: Losartan-treated, infected; UI [red]: untreated, infected). ( B ) Viral load in SARS-CoV-2-infected K18-hACE2 mice treated or not with Losartan or Losartan-Lisinopril combination on days 3, 5, or 6/7 post-infection. (*) Significant difference between groups: * p < 0.05, ** p < 0.005, **** p < 0.0001. The normality of the data was assessed using the Shapiro–Wilk test. A one-way ANOVA was performed, followed by the Bonferroni post-test (A-Treated non-infected), and mixed-effects analyses were conducted, followed by the Tukey post-test (A-Treated infected/B). (*) Significant difference between specific groups as indicated by the lines: * p < 0.05, ** p < 0.005, **** p < 0.0001. Data are presented as mean ± SEM. n = 10 per time point.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Enzyme-linked Immunosorbent Assay, Infection

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: Analysis of IL-6 and TNF-α in serum ( A , B ) and lung ( C , D ) of SARS-CoV-2-infected K18-hACE2 mice treated or not with Losartan or Losartan + Lisinopril combination at 6/7 dpi. CTI = Combination-treated Infected, LTI = Losartan-treated Infected, and UI = Untreated Infected. (*) Significant difference: * p < 0.05, ** p < 0.005 and *** p < 0.0005. Analysis was performed using ANOVA, followed by Bonferroni’s multiple comparisons test. Data presented as mean ± SEM. n = 10 mice per group.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: Histopathological analysis of lungs at days 6/7 post-infection of SARS-CoV-2-infected K18-hACE2 mice treated of not with Losartan or Losartan-Lisinopril combination. CTI = Combination-treated Infected, LTI = Losartan-treated Infected, and UI = Untreated Infected. ( A ) Results obtained from the sum of the degrees of intensity (maximum score of 44), *** p < 0.0005. Data are presented as individual scores and mean ± SD. The normality was assessed using the Shapiro–Wilk test. The Kruskal–Wallis test was used, followed by the Dunn post-test. ( B – D ) Representative histopathological images, emphasizing inflammatory infiltrate and thickness: ( B ) CTI-mice (score 1), ( C ) LTI-mice (score 1), and ( D ) UI-mice (score 4). ( E ) Histopathological findings, data presented as mean. n = 10 animals per group. Bar = 50 µm.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: Angiotensin-Converting Enzyme Inhibition and/or Angiotensin Receptor Blockade Modulate Cytokine Profiles and Improve Clinical Outcomes in Experimental COVID-19 Infection

doi: 10.3390/ijms26167663

Figure Lengend Snippet: ( A ) Survival curve of uninfected control (black line, n = 10) and SARS-CoV-2-infected K18-hACE2 mice untreated (red line, n = 20) or treated with Losartan (green line, n = 18) or Losartan-Lisinopril combination (gray line, n = 18). Untreated Uninfected did not show mortality. No significant differences in survival were observed between infected groups. The survival log-rank (Mantel–Cox) test was applied. ( B ) Analysis of weight loss of uninfected controls (black columns) or survivor SARS-CoV-2-infected K18-hACE2 mice untreated (red columns) or treated with Losartan (green columns) or Losartan-lisinopril (gray columns) at day 0 (pre-infection) and 15 days post-infection (dpi). ( C ) clinical scores of the experimental groups at 15 dpi. Normality was assessed using the Shapiro–Wilk test. Analysis was performed using ANOVA, followed by Bonferroni’s multiple comparisons test. (*) Significant differences: ** p < 0.005, *** p < 0.0005 and **** p < 0.0001.

Article Snippet: Detection and quantification of ACE2 protein in tissue samples were performed using the Human ACE2 DuoSet ELISA kit (R&D Systems- Cat. DY933-05, Minnneapolis, MN, USA), following the manufacturer’s recommendations.

Techniques: Control, Infection

Journal: The Journal of biological chemistry

Article Title: Neutrophil elastase decreases SARS-CoV-2 spike protein binding to human bronchial epithelia by clipping ACE-2 ectodomain from the epithelial surface.

doi: 10.1016/j.jbc.2023.104820

Figure Lengend Snippet: Figure 1. Quantitation of ACE-2 levels in plasma and airway mucus from subjects with and without CF and association of ACE-2 levels in CF sputum with neutrophil elastase activity. Blood samples collected from subjects with or without CF were processed for plasma collection, aliquoted, and stored at −80 C until further use. Frozen sputa from patients with CF or mucus from endotracheal tubes (ETTs) of healthy adult patients were mixed with NS containing 10% Sputolysin at 1:1 (sputum/mucus [mg]: volume [μl]) (37 C, 15 min), and sputum/mucus supernatants were collected by centrifugation. Plasma and sputum and mucus supernatants were analyzed for sACE-2 by ELISA. Data are summarized as (mean ± SEM) for plasma samples (20 non-CF and 16 CF) (A) and sputum/mucus samples (12 non-CF and 14 CF) (B). Statistical comparisons were made using Mann–Whitney U test. Soluble ACE-2 levels were significantly increased in CF sputum compared to non-CF, *p = 0.0146. For analysis of sputum NE activity compared to sputum ACE-2 levels, sputum supernatants prepared with DNase-1 (0.3 mg/ml) for 2 h at 37 C were used. Scatter dot plot showed a linear correlation between ACE-2 and NE activity levels in CF sputum supernatants (n = 20, r2 = 0.24, p = 0.03) (C). ACE-2, angiotensin-converting enzyme 2; CF, cystic fibrosis; NE, neutrophil elastase; NS, normal saline; sACE-2, soluble ACE-2.

Article Snippet: Levels of sACE-2 were quantified in equal volumes of sputum supernatants and plasma samples, using sandwich ELISA kit following manufacturer’s instructions (Catalog number DY933-05, Human ACE-2 DuoSet ELISA, R&D Systems).

Techniques: Quantitation Assay, Clinical Proteomics, Activity Assay, Centrifugation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Saline