adn  (R&D Systems)

Journal: Journal of Inflammation Research

Article Title: Unleashing AdipoRon’s Potential: A Fresh Approach to Tackle Pseudomonas aeruginosa Infections in Bronchiectasis via Sphingosine Metabolism Modulation

doi: 10.2147/JIR.S483689

Figure Lengend Snippet: The expression of adiponectin and AdipoR1 within bronchial epithelium determined in Pseudomonas aeruginosa infected mouse models. ( A ) Comparison of the expression of adiponectin between Pseudomonas aeruginosa infected mouse models and the uninfected mouse models by immunofluorescence (×200). ( B ) Comparison of the expression of AdipoR1 between Pseudomonas aeruginosa infected mouse models and the uninfected mouse models by immunofluorescence (×200). ( C ) Comparison of the expression of AdipoR1 between Pseudomonas aeruginosa infected mouse models and the uninfected mouse models by Western blot. ( D ) The expression levels of AdipoR1 in the bronchial epithelium of mice in different groups (DMSO control group, AdipoRon high-dose group, AdipoRon low-dose group and adiponectin Acrp30 recombinant protein group) were compared by immunofluorescence (×200). Data are expressed as the means ± standard deviation of three independent experiments. **indicates P <0.01.

Article Snippet: Treatment of the five groups: (i) Uninfected control: mice were anesthetized by intraperitoneal injection and instilled with 35 μL of PBS via the airway; (ii) Pseudomonas aeruginosa infection with DMSO control: 8 hours prior, mice were given an intravenous injection of DMSO solution as a control; (iii) Pseudomonas aeruginosa infection with low-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (5 mg/kg, intravenous injection); (iv) Pseudomonas aeruginosa infection with high-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (50 mg/kg, intravenous injection); and (v) Pseudomonas aeruginosa infection with adiponectin Acrp30 recombinant protein: 4 hours prior, mice were treated with adiponectin Acrp30 recombinant protein (HY-P7358, MedChemExpress, USA, 1 mg/kg, intravenous injection) as described.

Techniques: Expressing, Infection, Comparison, Immunofluorescence, Western Blot, Control, Recombinant, Standard Deviation

Journal: Journal of Inflammation Research

Article Title: Unleashing AdipoRon’s Potential: A Fresh Approach to Tackle Pseudomonas aeruginosa Infections in Bronchiectasis via Sphingosine Metabolism Modulation

doi: 10.2147/JIR.S483689

Figure Lengend Snippet: AdipoRon reduced the load of Pseudomonas aeruginosa in the airway of mice. ( A ) HE staining was performed on lung tissue specimens from mouse models with different treatment (control group without Pseudomonas aeruginosa infection, DMSO control group with Pseudomonas aeruginosa infection, AdipoRon low-dose (5 mg/kg) pretreatment group with Pseudomonas aeruginosa infection, AdipoRon high-dose (50 mg/kg) pretreatment group with Pseudomonas aeruginosa infection, and ACRP30 recombinant protein pretreatment group with Pseudomonas aeruginosa infection. ( B ) A semi-quantitative grading method of inflammation score was used to evaluate the severity of peribronchial inflammatory cell infiltration: 0, normal; 1, few cells; 2, a ring of inflammatory cells 1 cell layer deep; 3, a ring of inflammatory cells 2–4 cells deep; 4, a ring of inflammatory cells of > 4 cells deep. The inflammation scores of different groups were compared. ( C and D ) The CFU of Pseudomonas aeruginosa was counted in the lung tissue homogenates of mice in different treatment groups. ( E ) The bacterial load of Pseudomonas aeruginosa in the lung tissue of mice in different treatment groups was detected by immunofluorescence with the antibody against Pseudomonas aeruginosa (PA1-73116). Data are expressed as the means ± standard deviation of three independent experiments. **indicates P <0.01.

Article Snippet: Treatment of the five groups: (i) Uninfected control: mice were anesthetized by intraperitoneal injection and instilled with 35 μL of PBS via the airway; (ii) Pseudomonas aeruginosa infection with DMSO control: 8 hours prior, mice were given an intravenous injection of DMSO solution as a control; (iii) Pseudomonas aeruginosa infection with low-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (5 mg/kg, intravenous injection); (iv) Pseudomonas aeruginosa infection with high-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (50 mg/kg, intravenous injection); and (v) Pseudomonas aeruginosa infection with adiponectin Acrp30 recombinant protein: 4 hours prior, mice were treated with adiponectin Acrp30 recombinant protein (HY-P7358, MedChemExpress, USA, 1 mg/kg, intravenous injection) as described.

Techniques: Staining, Control, Infection, Recombinant, Immunofluorescence, Standard Deviation

Journal: Journal of Inflammation Research

Article Title: Unleashing AdipoRon’s Potential: A Fresh Approach to Tackle Pseudomonas aeruginosa Infections in Bronchiectasis via Sphingosine Metabolism Modulation

doi: 10.2147/JIR.S483689

Figure Lengend Snippet: AdipoRon increased sphingosine level in the lung of mouse infection model. ( A )The levels of sphingosine were compared between the Pseudomonas aeruginosa -infected mouse model group and the uninfected control group. ( B ) The metabolites in the lung tissue of mice in 3 groups were compared by PCA analysis: DMSO control group, adipoRon low-dose group and adipoRon high-dose group. ( C ) The levels of sphingosine in 3 groups were compared. ( D ) The metabolites in the lung tissue of mice were compared between adiponectin Acrp30 recombinant protein pretreatment group and the DMSO control group by PCA analysis. ( E ) The levels of sphingosine in 2 groups were compared. *** P <0.001.

Article Snippet: Treatment of the five groups: (i) Uninfected control: mice were anesthetized by intraperitoneal injection and instilled with 35 μL of PBS via the airway; (ii) Pseudomonas aeruginosa infection with DMSO control: 8 hours prior, mice were given an intravenous injection of DMSO solution as a control; (iii) Pseudomonas aeruginosa infection with low-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (5 mg/kg, intravenous injection); (iv) Pseudomonas aeruginosa infection with high-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (50 mg/kg, intravenous injection); and (v) Pseudomonas aeruginosa infection with adiponectin Acrp30 recombinant protein: 4 hours prior, mice were treated with adiponectin Acrp30 recombinant protein (HY-P7358, MedChemExpress, USA, 1 mg/kg, intravenous injection) as described.

Techniques: Infection, Control, Recombinant

Journal: Journal of Inflammation Research

Article Title: Unleashing AdipoRon’s Potential: A Fresh Approach to Tackle Pseudomonas aeruginosa Infections in Bronchiectasis via Sphingosine Metabolism Modulation

doi: 10.2147/JIR.S483689

Figure Lengend Snippet: AdipoRon activated P-AMPKα/PGC1α, inhibited TLR4/P-NF-κB p65, and reduced expression of bax in Pseudomonas aeruginosa -infected mouse model. ( A and B ) Western blot was used to quantitatively analyze the expression levels of several major indicators in the lung tissues of mice in three treatment groups (DMSO control group, AdipoRon low-dose group and AdipoRon high-dose group). Image J software was used to analyze the gray levels of different protein imprinting bands and to make statistical analysis. ( C and D ) Western blot was used to quantitatively analyze the expression levels of several major indexes in the lung tissues of mice in two groups: DMSO control group and Acrp30 recombinant protein pretreatment group. Data are expressed as the means ± standard deviation of three independent experiments. *indicates P <0.05, **indicates P <0.01.

Article Snippet: Treatment of the five groups: (i) Uninfected control: mice were anesthetized by intraperitoneal injection and instilled with 35 μL of PBS via the airway; (ii) Pseudomonas aeruginosa infection with DMSO control: 8 hours prior, mice were given an intravenous injection of DMSO solution as a control; (iii) Pseudomonas aeruginosa infection with low-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (5 mg/kg, intravenous injection); (iv) Pseudomonas aeruginosa infection with high-dose AdipoRon: 8 hours prior, mice were treated with AdipoRon (50 mg/kg, intravenous injection); and (v) Pseudomonas aeruginosa infection with adiponectin Acrp30 recombinant protein: 4 hours prior, mice were treated with adiponectin Acrp30 recombinant protein (HY-P7358, MedChemExpress, USA, 1 mg/kg, intravenous injection) as described.

Techniques: Expressing, Infection, Western Blot, Control, Software, Recombinant, Standard Deviation

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Cumulative concentration response curve of methylcholine-induced vasodilation in pre-constricted (with U46619) mesenteric arteries incubated overnight without stimulus (white), 50 µg/mL of oxLDL (red), 30 µg/mL of AN (green), or 30 µg/mL of AN plus 50 µg/mL of oxLDL (blue). (B) Summary graph of the maximum response (Emax) obtained from concentrations curves shown in (A). Vessels were obtained from rats ( n = 6-13). (C) Cumulative concentration response curve of methylcholine-induced vasodilation in pre-constricted (with U46619) mesenteric arteries incubated overnight without stimulus (white) or with 50 µg/mL of oxLDL (red), 30 µg/mL of AN plus 50 µg/mL of oxLDL (blue), 30 µg/mL of AN plus 10 µg/mL of TS20 (purple), or 10 µg/mL of the IgG isotype (black). (D) Summary graph of the maximum response (Emax) obtained from concentrations curves shown in C. Vessels from n = 6-7 rats. Values are shown as the mean±SEM. ＊ p ＜0.05; ＊＊ p ＜0.01; ＊＊＊ p ＜0.001. ns, non-significant.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Concentration Assay, Incubation

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Schematic representation of recombinant human T-cadherin proteins fused with Myc-His-tag. (B) Coomassie brilliant blue (CBB) staining and western blot analysis of purified T-cadherin proteins. (C) Binding of recombinant AN to immobilized T-cadherin in ELISA was determined by anti-AN antibody. (D) Binding of recombinant AN−oxLDL complex to immobilized LOX-1 in ELISA was determined by anti-AN antibody. s.p., signal peptide; EC, extracellular cadherin.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Staining, Western Blot, Purification, Binding Assay

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: Correlation between total-AN and high-molecular weight (HMW)-AN (left), total-AN and T-cadherin bindable adiponectin (Tcad-AN) (middle), HMW-AN and Tcad-AN (right).

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Enzyme-linked Immunosorbent Assay, High Molecular Weight

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: (A) Serum levels of AN, T-cadherin bindable adiponectin (Tcad-AN), oxidized LDL (oxLDL) and LOX-1-ligand containing apoB (LAB) in patients on hemodialysis (HD; n = 35) and control subjects ( n = 35). The error bars represent the mean±SD of the values obtained. (B) Comparison of the proportion of MAC-high between HD and controls. MAC-high and -low groups were divided by the median of the MAC values of 70 subjects.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Comparison, Control

Journal: Journal of Atherosclerosis and Thrombosis

Article Title: A Novel ELISA System for Measuring Modified LDL-Adiponectin Complex

doi: 10.5551/jat.65377

Figure Lengend Snippet: The error bars represent mean±SD.

Article Snippet: The wells were washed three times with PBS, and the plates were incubated for 2 h at 25°C with 20 μL of the recombinant adiponectin (BioVendor R&D, Brno, Czech Republic, RD172023100) or serum diluted four times with HEPES-NaCl buffer (10 mM HEPES, 150 mM NaCl, pH 7.4) containing 1 mM CaCl 2 and 0.1 mM MgCl 2 .

Techniques: Comparison