Journal: The Journal of International Medical Research

Article Title: Upregulation of matrix metalloproteinase 9 (MMP9)/tissue inhibitor of metalloproteinase 1 (TIMP1) and MMP2/TIMP2 ratios may be involved in lipopolysaccharide-induced acute lung injury

doi: 10.1177/0300060520919592

Figure Lengend Snippet: Primer sequences for real-time quantitative polymerase chain reaction analyses.

Article Snippet: Mouse anti-rat MMP9 (cat. no. NBP2-13173SS) and TIMP2 antibodies (cat. no. NBP1-42375) were purchased from Novus Biologicals Inc. (Littleton, CO, USA).

Techniques: Real-time Polymerase Chain Reaction

Journal: The Journal of International Medical Research

Article Title: Upregulation of matrix metalloproteinase 9 (MMP9)/tissue inhibitor of metalloproteinase 1 (TIMP1) and MMP2/TIMP2 ratios may be involved in lipopolysaccharide-induced acute lung injury

doi: 10.1177/0300060520919592

Figure Lengend Snippet: Lung MMP and TIMP mRNA and protein expression. Changes in MMP2 (a), TIMP2 (b), MMP2/TIMP2 (c), MMP9 (d), TIMP1 (e), and MMP9/TIMP1 (f) in all groups treated with or without LPS. (g) Protein expression in each group as detected by western blotting. LPS 2h, 6h, 12h, and 24h indicate 2, 6, 12, and 24 hours after LPS injection in the acute lung injury group, respectively. Data are shown as mean ± standard deviation (n = 6 per group). * P < 0.05 vs. control, # P < 0.05 vs. LPS 2h. Control, normal control group; MMP, matrix metalloproteinase; TIMP, tissue inhibitor of metalloproteinase; LPS, lipopolysaccharide

Article Snippet: Mouse anti-rat MMP9 (cat. no. NBP2-13173SS) and TIMP2 antibodies (cat. no. NBP1-42375) were purchased from Novus Biologicals Inc. (Littleton, CO, USA).

Techniques: Expressing, Western Blot, Injection, Standard Deviation, Control

Journal: Transplantation

Article Title: Urinary TIMP-2 Predicts the Presence and Duration of Delayed Graft Function in Donation After Circulatory Death Kidney Transplant Recipients

doi: 10.1097/tp.0000000000002472

Figure Lengend Snippet: FIGURE 2. TIMP-2 and IGFBP7 values were significantly different for patients with and without fDGF. Median ± IQR of both TIMP-2 and IGFBP7 are plotted for patients with and without fDGF. TIMP-2 values were significantly higher for patients with fDGF as compared to patients without fDGF (A). This also applied for the IGFBP7 values (B). Interestingly, TIMP-2 values of all patients decreased in the first 10 days after transplantation, where as IGFBP7 values increased. fDGF, functionally defined delayed graft function; IGFBP7, insulin-like growth factor-binding protein 7; IQR, interquartile range; TIMP-2, tissue inhibitor of metalloproteinases-2.

Article Snippet: Urinary TIMP-2 and IGFBP7 were quantified using sandwich immunoassays (ELISA, DTM200; R&D Systems, Minneapolis, MN for TIMP-2 and EK0991, Boster Biological Technology, Pleasanton, CA for IGFBP7, respectively) according to the manufacturer’s instructions.

Techniques: Transplantation Assay, Binding Assay

Journal: Transplantation

Article Title: Urinary TIMP-2 Predicts the Presence and Duration of Delayed Graft Function in Donation After Circulatory Death Kidney Transplant Recipients

doi: 10.1097/tp.0000000000002472

Figure Lengend Snippet: FIGURE 3. ROC curves of TIMP-2 and IGFBP7 in the prediction of fDGF and prolonged fDGF The absolute concentration of TIMP-2 on day 1 could adequately identify patients with fDGF, and this further improved in case TIMP-2 was corrected for osmolality (TIMP-2/ mOsm) (A). The concentration of IGFBP7 on day 1, on the other hand, showed only limited predictive value also when corrected for osmolality (IGFBP7/mOsm) (B). On day 10, both TIMP-2 and TIMP-2/mOsm could also adequately predict prolonged fDGF duration ≥21 days (C), whereas IGFBP7 and IGFBP7/mOsm could not (D). fDGF, functionally defined delayed graft function; IGFBP7, insulin-like growth factor-binding protein 7; ROC, receiver operating characteristic; TIMP-2, tissue inhibitor of metalloproteinases-2.

Article Snippet: Urinary TIMP-2 and IGFBP7 were quantified using sandwich immunoassays (ELISA, DTM200; R&D Systems, Minneapolis, MN for TIMP-2 and EK0991, Boster Biological Technology, Pleasanton, CA for IGFBP7, respectively) according to the manufacturer’s instructions.

Techniques: Concentration Assay, Binding Assay

Journal: Transplantation

Article Title: Urinary TIMP-2 Predicts the Presence and Duration of Delayed Graft Function in Donation After Circulatory Death Kidney Transplant Recipients

doi: 10.1097/tp.0000000000002472

Figure Lengend Snippet: FIGURE 5. TIMP-2/mOsm and IGFBP7/mOsm in tertiles. Patients were divided into tertiles based on TIMP-2/mOsm and IGFBP7/ mOsm concentrations, and the percentage of patients with fDGF and prolonged fDGF in each tertile was calculated. On day 1, patients without fDGF predominantly grouped together in the tertile with the lowest values for TIMP-2/mOsm, (77%), whereas patients with fDGF (92%) were in the middle or highest tertile (A). For IGFBP7/mOsm, a comparable figure was seen, with slightly lower percentages (B). On day 10, patients with prolonged fDGF predominantly grouped together in the tertile with the highest TIMP-2/mOsm values (83.3%), and none had TIMP-2/mOsm values in the lowest tertile (C). In contrast, patients with prolonged fDGF did not group together according to their IGFBP7/mOsm values (D). fDGF, functionally defined delayed graft function; IGFBP7, insulin-like growth factor-binding protein 7; TIMP-2, tissue inhibitor of metalloproteinases-2.

Article Snippet: Urinary TIMP-2 and IGFBP7 were quantified using sandwich immunoassays (ELISA, DTM200; R&D Systems, Minneapolis, MN for TIMP-2 and EK0991, Boster Biological Technology, Pleasanton, CA for IGFBP7, respectively) according to the manufacturer’s instructions.

Techniques: Binding Assay

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