Journal: International Journal of Medical Sciences

Article Title: Comparison the Prognostic Value of Galectin-3 and Serum Markers of Cardiac Extracellular Matrix Turnover in Patients with Chronic Systolic Heart Failure

doi: 10.7150/ijms.8083

Figure Lengend Snippet: Receiver operating characteristic (ROC) curves for prediction mortality. The area under the curve (AUC) of log MMP-2, log TIMP-1, log BNP, log PIIINP, log Gal-3, log PINP, and log MMP-9 were 0.786, 0.699, 0.636, 0.625, 0.607, 0.519, and 0.367, respectively. Abbreviations: BNP= brain natriuretic peptide; Gal-3= Galectin-3; HF=heart failure; LVEF= left ventricular ejection fraction; MMP = matrix metalloproteinase; PINP = type I amioterminal propeptide of procollagen; PIIINP = type III amioterminal propeptide of procollagen; TIMP = tissue inhibitor of metalloproteinase.

Article Snippet: TIMP-1 was measured by an ELISA kit (DTM100, R & D systems, Minneapolis, USA).

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Comparison the Prognostic Value of Galectin-3 and Serum Markers of Cardiac Extracellular Matrix Turnover in Patients with Chronic Systolic Heart Failure

doi: 10.7150/ijms.8083

Figure Lengend Snippet: Kaplan-Meier analysis of cumulative rates of survival in HF patients with higher or lower levels of serum Gal-3 or cardiac ECM markers. The p value of Gal-3, PINP, PIIINP, TIMP-1, MMP-2, MMP-9, and BNP were 0.153, 0.708, 0.154, 0.028, 0.001, 0.501, and 0.483, respectively. Abbreviations: BNP= brain natriuretic peptide; ECM= Extracellular matrix; Gal-3= Galectin-3; HF=heart failure; LVEF= left ventricular ejection fraction; MMP = matrix metalloproteinase; PINP = type I amioterminal propeptide of procollagen; PIIINP = type III amioterminal propeptide of procollagen; TIMP = tissue inhibitor of metalloproteinase.

Article Snippet: TIMP-1 was measured by an ELISA kit (DTM100, R & D systems, Minneapolis, USA).

Techniques:

Journal: International Journal of Medical Sciences

Article Title: Comparison the Prognostic Value of Galectin-3 and Serum Markers of Cardiac Extracellular Matrix Turnover in Patients with Chronic Systolic Heart Failure

doi: 10.7150/ijms.8083

Figure Lengend Snippet: Kaplan-Meier analysis of cumulative rates of HF admission-free survival in HF patients with higher or lower levels of serum Gal-3 or cardiac ECM markers. The p value of Gal-3, PINP, PIIINP, TIMP-1, MMP-2, MMP-9, and BNP were 0.166, 0.624, 0.639, 0.085, <0.001, 0.624, and 0.684, respectively. Abbreviations: BNP= brain natriuretic peptide; ECM= Extracellular matrix; Gal-3= Galectin-3; HF=heart failure; MMP = matrix metalloproteinase; PINP = type I amioterminal propeptide of procollagen; PIIINP = type III amioterminal propeptide of procollagen; TIMP = tissue inhibitor of metalloproteinase.

Article Snippet: TIMP-1 was measured by an ELISA kit (DTM100, R & D systems, Minneapolis, USA).

Techniques:

Journal: The Journal of biological chemistry

Article Title: The C-terminal domains of ADAMTS1 contain exosites involved in its proteoglycanase activity.

doi: 10.1016/j.jbc.2023.103048

Figure Lengend Snippet: Figure 2. Inhibition of ADAMTS1 by TIMPs. A, inhibition of versicanase activity by TIMP family members. TIMP1, TIMP2, TIMP3, and TIMP4 (each at 500 nM) were incubated with ADAMTS1 (100 nM) for 1 h at 37 C before addition of V1-5GAG and digestion for 2 h. Following SDS-PAGE under reducing conditions (5% β-mercaptoethanol) and immunoblotting, FL V1-5GAG and versikine (VSK) were detected by the anti-Vc antibody. A representative immunoblot is shown (n = 2 independent experiments). B, inhibition of peptidolytic activity. TIMPs (each 25 nM) were incubated with a nominal con- centration of 25 nM ADAMTS1 for 1 h at 37 C before addition of the QF peptide substrate fluorescein-5(6)-carbonyl-Ala-Glu-Leu-Asn-Gly-Arg-Pro-Ile-Ser-Ile- Ala-Lys (5(6)-TAMRA) (3.5 μM) and digestion for 2 h. Following subtraction of the background (reactions not containing ADAMTS1), values were converted into percentage of ADAMTS1 activity in the absence of TIMPs and reported as average ± SD (n = 3, each point representing a technical replicate), p < 0.05 by Mann-Whitney test. C, titration of ADAMTS1 with TIMP3. TIMP3 (0–16 nM) was incubated with ADAMTS1 (20 nM nominal concentration) at 37 C for 1 h, and residual activity against the QF peptide FAM-AELNGRPISIAK-Tamra (3.5 μM) was determined. A representative titration curve is shown, each point representing a mean of two technical replicates. Final concentration of ADAMTS1 following titration was 10 nM. FL, full-length; No E, no enzyme; No I, no inhibitor; QF, Quenched-Fluorescent; TIMP, tissue inhibitor of metalloproteinase.

Article Snippet: Semiquantitative proteoglycan cleavage assays Purified V1-5GAG (100 nM) was digested with ADAMTS1, in TNC-B buffer at 37 C for 2 h. Where indicated, 500 μM recombinant human TIMP1, TIMP2, TIMP3, or TIMP4 (R&D Systems, Cat. n.: 970-TM, 971-TM, 973-TM, 974-TSF) were preincubated with 100 nM ADAMTS1 for 1 h at 37 C before digestion.

Techniques: Inhibition, Activity Assay, Incubation, SDS Page, Western Blot, MANN-WHITNEY, Titration, Concentration Assay