Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Plasma sRAGE and Ang-2 levels stratified by PARDS severity Comparison was made between mild, moderate and severe PARDS versus control using the Mann-Whitney U test. P values are subjected to Bonferroni correction for multiple testing. Ang-2, angiopoietin-2; PARDS, paediatric acute respiratory distress syndrome; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics, Comparison, Control, MANN-WHITNEY

Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Correlation matrix of plasma sRAGE and Ang-2 with age, PIM 3 score, PELOD2 score and the oxygenation index. Spearman test was used to assess correlation. Numbers in cells refer to Spearman’s correlation coefficient. *p<0.05, **p<0.01, ***p<0.001. Ang-2, angiopoietin-2; PELOD 2, Pediatric Logistic Organ Dysfunction 2 score; PIM 3, Pediatric Index of Mortality 3 score; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics

Journal: BMJ Open Respiratory Research

Article Title: Validation of plasma soluble receptor of advanced glycation end-products and angiopoietin-2 in paediatric acute respiratory distress syndrome

doi: 10.1136/bmjresp-2025-003630

Figure Lengend Snippet: Plasma sRAGE and Ang-2 levels in the presence of cardiovascular dysfunction, multiorgan dysfunction and mortality. Data presented as median (IQR), and comparison was made using the Mann-Whitney test. Organ dysfunctions were defined according to the International Pediatric Sepsis Consensus Conference. Ang-2, angiopoietin-2; ICU, intensive care unit; sRAGE, soluble receptor for advanced glycation end-products.

Article Snippet: The plasma was then aspirated and stored at −80°C for later batched analysis. sRAGE (R&D Systems; catalogue no. DY1145) and Ang-2 (R&D Systems; catalogue no. DY623) were measured in duplicate using an ELISA according to manufacturer’s protocol.

Techniques: Clinical Proteomics, Comparison, MANN-WHITNEY

Journal: medRxiv

Article Title: Aerobic Exercise Training Elevates Circulating sRAGE via Modulation of Sheddase Regulation in Adults with Type 2 Diabetes

doi: 10.1101/2025.09.10.25335517

Figure Lengend Snippet: Basal solubilized receptor for advanced glycation end products (sRAGE; A), solubilized Toll-like receptor 4 (sTLR4, C) pre and post 12-week intervention in the control (CON) and aerobic exercise training (AE) groups. Respective absolute changes in basal sRAGE (B) and sTLR4 (C) and percentage changes in sRAGE (E) and sTLR4 (E) were calculated. *P < 0.05 vs Pre, # P < 0.05 vs CON. Statistical analyses performed include mixed model regression analyses with Bonferroni post hoc (panels A & C) and two-tailed t-test (panels B, D, & E).

Article Snippet: sRAGE was quantified via sRAGE ELISA (DRG000, R&D Systems, Minneapolis, MN) according to manufacturer’s instructions.

Techniques: Control, Two Tailed Test

Journal: PLoS ONE

Article Title: Beyond Genetic Factors in Familial Amyloidotic Polyneuropathy: Protein Glycation and the Loss of Fibrinogen's Chaperone Activity

doi: 10.1371/journal.pone.0024850

Figure Lengend Snippet: (A) Western blot analysis of TTR in the plasma from three control and three FAP individuals after aggregation induced by temperature. (B) Western blot analysis of TTR in the plasma from three FAP individuals after aggregation induced by temperature in the absence of fibrinogen, in the presence of glycated fibrinogen and in the presence of non-glycated fibrinogen. (C) Western blot analysis of fibrinogen in the plasma from three control and three FAP individuals, Western blot of sRAGE was used as loading control. (D) Coomassie blue stained gel of α-crystallin and fibrinogen without glycation and glycated with 2.5 and 10 mM of methylglyoxal. Western blot analysis of argpyrimidine modified proteins. (E) Chemically (DTT) induced protein aggregation of insulin and lysozyme (The aggregation of insulin and lysozyme alone, respectively, are used as controls) in the presence of α-crystallin and fibrinogen with and without glycation. Error bars indicate standard deviation.

Article Snippet: Antibodies used were: anti-human TTR polyclonal antibody (Santa Cruz Biotechnology) at a dilution of 1∶5000; anti-argpyrimidine (Jaica) at a dilution of 1∶2000; anti-human soluble RAGE (Santa Cruz Biotechnology) at a dilution of 1∶5000; anti-fibrinogen (Calbiochem) at a dilution of 1∶10000.

Techniques: Western Blot, Clinical Proteomics, Control, Staining, Modification, Standard Deviation

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition restores alveolar fluid clearance after acid-induced lung injury. Measurement of net alveolar fluid clearance (AFC) rate as a marker of epithelial function in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. P = 0.02 for the overall multiple group test; no statistical difference was found in post-hoc tests.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Marker, Recombinant, Whisker Assay

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition improves arterial oxygenation after acid-induced lung injury. Arterial oxygen tension (PaO 2 )/inspiratory oxygen fraction (FiO 2 ) in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Individual values are reported. Time × group interaction and post-hoc comparisons were verified using random effects models to analyse longitudinal evolution of variables.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition decreases alveolar-capillary permeability after acid-induced lung injury. Values are reported as box and whisker plots. ( A ) Level of total protein in the bronchoalveolar lavage (BAL) fluid from uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. ( B ) Extravascular lung water, as measured by transpulmonary thermodilution (Picco + , Pulsion SA) and indexed to body weight, in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group at each time point). Time × group interaction and post-hoc comparisons were verified using random effects models to analyse longitudinal evolution of variables.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Permeability, Whisker Assay, Recombinant

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: RAGE inhibition decreases alveolar inflammation after acid-induced lung injury. Measurement of bronchoalveolar lavage (BAL) levels of ( A ) tumour necrosis factor (TNF)-α, ( B ) interleukin (IL)-6, ( C ) IL-1β and ( D ) IL-18 in uninjured (Sham), acid-injured (HCl) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant, Whisker Assay

Journal: Scientific Reports

Article Title: Inhibition of the Receptor for Advanced Glycation End-Products in Acute Respiratory Distress Syndrome: A Randomised Laboratory Trial in Piglets

doi: 10.1038/s41598-019-45798-5

Figure Lengend Snippet: (A ) RAGE inhibition decreases histological features of lung injury. Lung injury scores were higher in acid-injured (HCl) than in uninjured piglets (Sham) and acid-injured piglets treated with RAGE antagonist peptide (HCl + RAP) or recombinant sRAGE (HCl + sRAGE) (n = 12 per group). Values are reported as box and whisker plots. The Kruskal-Wallis test, with post-hoc Bonferroni test for pairwise comparisons were used. (B – E) Representative hematoxylin and eosin–stained sections at x20 original magnification of (B) uninjured piglets (Sham), (C) acid-injured piglets (HCl), acid-injured piglets treated with (D) RAGE antagonist peptide (HCl + RAP) or (E) recombinant sRAGE (HCl + sRAGE). There was greater cellularity consisting mainly of neutrophils (black arrowheads), more alveolar wall thickening (white arrowheads), and more areas of atelectasis and increased alveolar disruption, proteinous debris, and hemorrhage (black arrows) in untreated, acid-injured than in uninjured or treated piglets. Scale bars, 100 µm.

Article Snippet: The “sRAGE group” (n = 12) included animals with HCl-induced lung injury that also received intravenous treatment with sRAGE (3 mg.kg −1 ) (Recombinant Human RAGE Fc Chimera, R&D Systems, Minneapolis, MN).

Techniques: Inhibition, Recombinant, Whisker Assay, Staining, Disruption

Journal: Investigative Ophthalmology & Visual Science

Article Title: Advanced Glycation End Products and Receptor (RAGE) Promote Wound Healing of Human Corneal Epithelial Cells

doi: 10.1167/iovs.61.3.14

Figure Lengend Snippet: Involvement of the coupled RAGE ligand/receptor in human corneal epithelial (HCE) cell wound healing. Representative images of scratch assays performed on HCE cells (left panel) treated with HMGB1 (high mobility group box 1, 100 ng/mL) ( A ) or AGEs (advanced glycation end products) (10/100/200 µg/mL) (B , C ). Percentage of residual wound area (right panel) after treatment with HMGB1 (100 ng/mL) ( A ) or AGEs (10/100/200 µg/mL) ( B , C ), compared with 0 hours and standardized to the untreated condition (100%) (n = 5 experiments, each conducted in duplicate). ( D ) Representative images of scratch assays performed on HCE cells transiently transfected with siRNA against RAGE (siRNA RAGE) or siRNA control (Scramble) (100 nM) for 36 hours and then treated with AGEs (100 µg/mL) (left panel). Percentage of the residual wound area of HCE cells transfected with siRNA against RAGE (100 nM) for 36 hours and then treated with AGEs (100 µg/mL), compared with 0 h (right panel) (n = 5 experiments, each conducted in duplicate). Each bar graph shows mean ± SEM. Mann-Whitney test after a nonparametric ANOVA analysis; * P < 0.05; ** P < 0.01; *** P < 0.005; ns: not significant.

Article Snippet: Cells were transfected with 1 µg RAGE expression vector (RG204664; Origene, Herford, Germany) in the presence of 125 µL opti-MEM 1× and 5 µL p3000 reagent, mixed with 3.75 µL Lipofectamine 3000 reagent in 125 µL of opti-MEM (1×).

Techniques: Transfection, MANN-WHITNEY

Journal: Investigative Ophthalmology & Visual Science

Article Title: Advanced Glycation End Products and Receptor (RAGE) Promote Wound Healing of Human Corneal Epithelial Cells

doi: 10.1167/iovs.61.3.14

Figure Lengend Snippet: Functionality of the RAGE pathway. Characterization of the RAGE ( A ) mRNA and ( B ) protein expression in human cornea, primary human epithelial cells (mRNA only), and the HCE cell line (mRNA and protein) evaluated by ( A ) RT-PCR, ( B ) immunofluorescence, and ( B ) western blotting. For RT-PCR, negative controls (NC) were performed ( A ) without cDNA. ( B , left panel) Representative images of RAGE expression ( green ) in human corneas (top panel) and HCE cells (bottom panel). Nuclei were stained with Hoechst ( blue ); NC (left) were obtained by incubating HCE cells without primary antibody. ( B , right panel) Western blot experiments identified the RAGE protein at the described molecular weight (46 kDa). ( C ) Functionality of the NF-κB pathway (by luciferase reporter gene activity) was measured after treatment of HCE cells with AGEs (100 µg/mL) for 45 minutes (n = 5 experiments, each conducted in duplicate) (right panel). Positive controls (T+) were obtained by co-transfection with pMEKK (n = 5 experiments, each conducted in duplicate) (left panel). Each bar graph shows mean ± SEM. Mann-Whitney; * P < 0.05.

Article Snippet: Cells were transfected with 1 µg RAGE expression vector (RG204664; Origene, Herford, Germany) in the presence of 125 µL opti-MEM 1× and 5 µL p3000 reagent, mixed with 3.75 µL Lipofectamine 3000 reagent in 125 µL of opti-MEM (1×).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Immunofluorescence, Western Blot, Staining, Molecular Weight, Luciferase, Activity Assay, Cotransfection, MANN-WHITNEY

Journal: Investigative Ophthalmology & Visual Science

Article Title: Advanced Glycation End Products and Receptor (RAGE) Promote Wound Healing of Human Corneal Epithelial Cells

doi: 10.1167/iovs.61.3.14

Figure Lengend Snippet: Cx43 expression is induced by the AGEs/RAGE axis in HCE cells. Characterization of Cx43 protein expression ( arrow ) by immunostaining (bottom panel). Cells incubated without primary antibody served as a negative control (NC). ( B ) Relative quantification of Cx43 mRNA expression in untreated HCE cells 0, 6, and 24 hours after scratch wounding. Results were expressed as a ratio of the 0-hour condition (n = 5 experiments, each conducted in duplicate). ( C ) Quantification of Cx43 mRNA expression in HCE cells treated with AGEs (100 µg/mL) for 6 hours or 24 hours without scratch wounding (left panel) or with scratch wounding (right panel). Cells not treated with AGEs served as a control. Results were expressed as a ratio of the untreated condition at each time point (n = 5 experiments, each conducted in duplicate). (Left panel) Quantification of Cx43 mRNA expression in HCE cells transiently transfected with siRNA against RAGE (siRNA RAGE) (100 nM) for 36 hours and then treated with AGEs (100 µg/mL) for 6 hours. Results were expressed as a ratio of the scrambled siRNA condition (n = 5 experiments, each conducted in duplicate). Each bar graph shows mean ± SEM. Mann-Whitney after a nonparametric ANOVA analysis; * P < 0.05; ns: not significant. ( D ) Relative quantification of Cx43 protein expression in HCE cells following treatment with AGEs (100 µg/mL) after scratch wounding. Untreated cells served as a control. Results were expressed as a ratio of the 0-hour, unwounded condition (n = 5 experiments) (top panel). Quantification of Cx43 protein expression in HCE cells treated with AGEs (100 µg/mL) for 12 and 24 hours after scratch wounding (bottom panel). Results were expressed as a ratio of the untreated condition at each time point (n = 5 experiments, each conducted in duplicate) (** P < 0.01). (E) Characterization by immunostaining of Cx43 protein expression according to the distance from the wound, time, and treatment with AGEs (100 µg/mL). Staining of Cx43 protein expression in HCE cells treated near the wound margins (left panel) and behind the wound (right panel). Cells not treated with AGEs (100 µg/mL) served as a control. Cells incubated without primary antibody served as a negative control (NC).

Article Snippet: Cells were transfected with 1 µg RAGE expression vector (RG204664; Origene, Herford, Germany) in the presence of 125 µL opti-MEM 1× and 5 µL p3000 reagent, mixed with 3.75 µL Lipofectamine 3000 reagent in 125 µL of opti-MEM (1×).

Techniques: Expressing, Immunostaining, Incubation, Negative Control, Transfection, MANN-WHITNEY, Staining