Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab and s-CSF2pep-Ab levels in patients with AIS and TIA and in HDs. (A–B) Levels of serum antibodies against (A) s-CSF2-Ab and (B) s-CSF2pep-Ab determined using amplified luminescence proximity homogeneous assay-linked immunosorbent assay (AlphaLISA), calculated by subtracting the levels of antibodies against the control GST. Scatter dot plots of the s-CSF2-Ab and s-CSF2pep-Ab levels are also presented. Bars represent averages ± standard deviation (SD). ** P < 0.05; * P < 0.01; ns, not significant, Kruskal–Wallis test. (C–E) A receiver operating characteristic (ROC) curve analysis was conducted to assess the ability of s-CSF2-Ab to detect AIS (C) and that of s-CSF2pep-Ab to detect AIS (D) and TIA (E) . The numbers indicate the cutoff values for the indicated markers, and the numbers in parentheses indicate sensitivity (left) and specificity (right). The areas under the ROC curve (AUC) and 95% confidence intervals (CIs) are also shown. AIS, acute ischemic stroke; HD, healthy donor; CSF2-Ab, CSF2 antibody; CSF2pep-Ab, CSF2 peptide antibody; TIA, transient ischemic attack.

Article Snippet: The reaction mixture containing 2.5 μL of serum samples diluted at 1:100 in AlphaLISA buffer (25 mM HEPES pH 7.4, 0.1% casein, 0.5% Triton X-100, 1 mg/mL of dextran-500, and 0.05% Proclin-300) and 2.5 μL of GST or GST-CSF2 proteins (10 μg/ml) or 400 ng/mL of bCSF2-77 was incubated in 384-well white opaque OptiPlate microtiter plates (PerkinElmer, Beaconsfield, United Kingdom) at room temperature for 6–8 h. Next, 2.5 μL of anti-human immunoglobulin G (IgG)-conjugated acceptor beads (40 μg/ml) and 2.5-μL glutathione-conjugated donor beads (40 μg/ml) or 2.5 μL of streptavidin-conjugated donor beads (40 μg/ml) were added.

Techniques: Comparison, Amplification, Control, Standard Deviation

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Comparison of the s-CSF2-Ab and s-CSF2pep-Ab levels between HDs and patients with AMI and DM. ( A,B ) The levels of s-CSF2-Ab and s-CSF2pep-Ab in HDs and patients with acute myocardial infarction (AMI) ( A ) and diabetes mellitus (DM) ( B ) were determined using AlphaLISA. The bars represent averages ± SD. *** P < 0.001, Mann–Whitney U test. ( D,E ) The ROC curves to assess the ability of s-CSF2-Ab to predict AMI and DM are presented in ( D ) and ( E ), respectively. ( F,G ) The ability of s-CSF2pep-Ab to predict AMI ( E ) and DM ( F ) was also evaluated via ROC analysis. Comparison of the overall survival in patients with DM between the s-CSF2-Ab positive (s-CSF2-Ab + ) and s-CSF2-Ab negative (s-CSF2-Ab − ) groups ( P = 0.019; C ) and between the s-CSF2pep-Ab positive (s-CSF2-Ab + ) and s-CSF2pep-Ab negative (s-CSF2-Ab − ) groups ( P = 0.034; C ). Statistical analyzes were performed by the Log-Rank test between two groups.

Article Snippet: The reaction mixture containing 2.5 μL of serum samples diluted at 1:100 in AlphaLISA buffer (25 mM HEPES pH 7.4, 0.1% casein, 0.5% Triton X-100, 1 mg/mL of dextran-500, and 0.05% Proclin-300) and 2.5 μL of GST or GST-CSF2 proteins (10 μg/ml) or 400 ng/mL of bCSF2-77 was incubated in 384-well white opaque OptiPlate microtiter plates (PerkinElmer, Beaconsfield, United Kingdom) at room temperature for 6–8 h. Next, 2.5 μL of anti-human immunoglobulin G (IgG)-conjugated acceptor beads (40 μg/ml) and 2.5-μL glutathione-conjugated donor beads (40 μg/ml) or 2.5 μL of streptavidin-conjugated donor beads (40 μg/ml) were added.

Techniques: Comparison, MANN-WHITNEY

Journal: Frontiers in Cardiovascular Medicine

Article Title: Utility of atherosclerosis-associated serum antibodies against colony-stimulating factor 2 in predicting the onset of acute ischemic stroke and prognosis of colorectal cancer

doi: 10.3389/fcvm.2023.1042272

Figure Lengend Snippet: Association of the s-CSF2-Ab and s-CSF2pep-Ab levels with CKD. The levels of s-CSF-Ab ( A ) and s-CSF2pep-Ab ( B ) in HDs and in patients with diabetic CKD (CKD type 1), nephrosclerosis (CKD type 2), and glomerulonephritis (CKD type 3) were examined using AlphaLISA. The bars represent averages ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001, Kruskal–Wallis test. The data are summarized in . ( C–E ) ROC analysis to determine the ability of s-CSF2-Ab to predict CKD type 1 ( C ), type 2 ( D ), and type 3 ( E ). ( F–H ) ROC analysis to determine the ability of s-CSF2pep-Ab to predict CKD type 1 ( F ), type 2 ( G ), and type 3 ( H ). The numbers in the graphs are the same as those presented . CKD, chronic kidney disease.

Article Snippet: The reaction mixture containing 2.5 μL of serum samples diluted at 1:100 in AlphaLISA buffer (25 mM HEPES pH 7.4, 0.1% casein, 0.5% Triton X-100, 1 mg/mL of dextran-500, and 0.05% Proclin-300) and 2.5 μL of GST or GST-CSF2 proteins (10 μg/ml) or 400 ng/mL of bCSF2-77 was incubated in 384-well white opaque OptiPlate microtiter plates (PerkinElmer, Beaconsfield, United Kingdom) at room temperature for 6–8 h. Next, 2.5 μL of anti-human immunoglobulin G (IgG)-conjugated acceptor beads (40 μg/ml) and 2.5-μL glutathione-conjugated donor beads (40 μg/ml) or 2.5 μL of streptavidin-conjugated donor beads (40 μg/ml) were added.

Techniques: