Journal: Renal Failure

Article Title: ADAMTS13 ameliorates diabetic nephropathy by Nrf2/GPX4/eNOS signaling pathway

doi: 10.1080/0886022X.2026.2646089

Figure Lengend Snippet: ADAMTS13 maintained renal function and attenuated renal fibrosis in vivo . Control mice were treated with vehicle (Control). The STZ-treated mice were infused with vehicle (DN) or rhADAMTS13 (DN + rhADAMTS13). rhADAMTS13 (2.6 ug/kg body weight) was injected into the tail vein daily for the subsequent 7 d. (A) The workflow of animal experiments. (B) Representative renal immunohistochemical staining of ADAMTS13 in the kidney. Scale Bar: 30 μm. (C) Representative renal morphological images of mice were shown. Hematoxylin-Eosin (HE), Masson’s trichrome and Periodic Acid-Schiff (PAS) staining of kidney slices. Scale Bar: 30 μm. (D) Body weight. (E) Serum glucose. (F) BUN. (G) Scr. (H) Urinary volume. (I) Proteinuria. (J) Urinary KIM-1. (K) Renal KIM-1 mRNA expression. ADAMTS13: a disintegrin and metalloprotease with a thrombospondin type 1 motif member 13; DN: diabetic nephropathy; BUN: blood urea nitrogen; Scr: serum creatinine; KIM-1: kidney injury molecule-1. Results were presented as mean ± SEM. N = 5, ** P < 0.01, *** P < 0.001, one-way ANOVA followed by Tukey’s post hoc test.

Article Snippet: Hyperglycemia was defined as a fasting blood glucose ≥11.1 mmol/L or random blood glucose ≥16.7 mmol/L. rhADAMTS13 (4245-AD, R&D Systems, Minneapolis, MN) was dissolved in saline and administered to mice via tail vein injections.

Techniques: In Vivo, Control, Injection, Immunohistochemical staining, Staining, Expressing

Journal: Blood Advances

Article Title: Development of a protease-resistant ADAMTS13 to improve stability against proteolytic degradation ∗

doi: 10.1182/bloodadvances.2024015212

Figure Lengend Snippet: ADAMTS13 degradation by purified proteases and in plasma-based assays. (A) Commercial rADAMTS13 (200 nM) was incubated with various proteases (100 nM) for 2 hours at 37°C. Samples were separated under reducing conditions using SDS-PAGE, and cleavage was visualized using SYPRO Ruby. (B) rADAMTS13, varying dilutions of tissue factor (TF), and 1 mg/mL GPRP-amide were incubated in human platelet-poor plasma for 15 minutes. Thrombin generation was then initiated with calcium and quantified over time in a parallel plate by measuring fluorescence (λ ex = 360 nm; λ em = 460) at 37°C. Curves were generated using the Technothrombin TGA Software in triplicate, and representative images are shown. (C) rADAMTS13 and varying concentrations of tPA were added to human platelet-poor plasma, followed by TF and calcium. Clot formation and lysis was quantified by measuring OD at 405 nm every 30 seconds in triplicate, and representative images are shown. Samples were separated via sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions, and ADAMTS13 degradation was visualized via western blot using an anti-ADAMTS13 metalloprotease domain antibody. Molecular weight references are indicated on the left of blots (kDa), and full-length bands are indicated by the black arrows. APC, activated protein C; GPRP-amide, Gly-Pro-Arg-Pro-amide; OD, optical density.

Article Snippet: Recombinant (r)ADAMTS13 and human ADAMTS13 Quantikine enzyme-linked immunosorbent assay kit (R&D Systems, Minneapolis, MN), kallikrein, FXIIa (Enzyme Research Laboratories, South Bend, IN), thrombin, FXa, FIXa, FXIa, FVIIa, activated protein C (APC), elastase, cathepsin G (Sigma-Aldrich, St Louis, MO), proteinase 3 (Enzo Life Sciences, Inc, Farmingdale, NY), FRETS-VWF73 (AnaSpec, Fremont, CA), AEBSF (4-[2-aminoethyl]benzenesulfonyl fluoride hydrochloride), Sivelestat (Tocris Bioscience, Bristol, United Kingdom), and cathepsin G inhibitor 1 (Cayman Chemical Company, Ann Arbor, MI).

Techniques: Purification, Clinical Proteomics, Incubation, SDS Page, Fluorescence, Generated, Software, Lysis, Polyacrylamide Gel Electrophoresis, Western Blot, Molecular Weight

Journal: Blood Advances

Article Title: Development of a protease-resistant ADAMTS13 to improve stability against proteolytic degradation ∗

doi: 10.1182/bloodadvances.2024015212

Figure Lengend Snippet: ADAMTS13 degradation by activated neutrophils. (A) ADAMTS13 was incubated with 0 × 10 3 to 500 × 10 3 PMA-stimulated neutrophils for 1 hour at 37°C, in the absence or presence of 2 mM AEBSF or 5 mM EDTA (B). (C) ADAMTS13 was incubated with 5 × 10 3 PMA-stimulated neutrophils with increasing concentrations of AEBSF, cathepsinG inhibitor I (CGI I), or Sivelestat for 1 hour. Samples were separated via SDS-PAGE and analyzed by (A) western blot using an anti-ADAMTS13 metalloprotease domain antibody, or (B-C) SYPRO Ruby. Black arrows indicate the level of full-length ADAMTS13 (∼180 kDa). AEBSF, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride; EDTA, ethylenediaminetetraacetic acid.

Article Snippet: Recombinant (r)ADAMTS13 and human ADAMTS13 Quantikine enzyme-linked immunosorbent assay kit (R&D Systems, Minneapolis, MN), kallikrein, FXIIa (Enzyme Research Laboratories, South Bend, IN), thrombin, FXa, FIXa, FXIa, FVIIa, activated protein C (APC), elastase, cathepsin G (Sigma-Aldrich, St Louis, MO), proteinase 3 (Enzo Life Sciences, Inc, Farmingdale, NY), FRETS-VWF73 (AnaSpec, Fremont, CA), AEBSF (4-[2-aminoethyl]benzenesulfonyl fluoride hydrochloride), Sivelestat (Tocris Bioscience, Bristol, United Kingdom), and cathepsin G inhibitor 1 (Cayman Chemical Company, Ann Arbor, MI).

Techniques: Incubation, SDS Page, Western Blot

Journal: Blood Advances

Article Title: Development of a protease-resistant ADAMTS13 to improve stability against proteolytic degradation ∗

doi: 10.1182/bloodadvances.2024015212

Figure Lengend Snippet: Characterizing the resistance of the ADAMTS13 mutants against proteolysis by purified proteases and in plasma fibrinolysis. (A) Four mutants of ADAMTS13 were developed: 2 with 1 of each C-terminal linker region mutated (T4L and T8L mutants), 1 with both linker regions mutated (T4L/T8L mutant), and a fourth with both linker regions and an additional elastase cleavage site mutation (T4L/T8L[I380G] mutant). (B) Using commercial recombinant ADAMTS13 (WT) as a control, each linker mutant (T4L, T8L, and/or T4L/T8L) was incubated at 50 nM with purified coagulation and fibrinolytic proteases at 50 nM (plasmin, thrombin, and kallikrein) or 100 nM (FXIa) for 0 to 180 minutes at 37°C. (C) The T4L/T8L mutant and 10 nM tPA were added to human platelet-poor plasma, followed by tissue factor and calcium at 37°C. Aliquots were removed at 0 and 45 minutes. (D) Mutants were incubated with 50 nM purified neutrophil-derived proteases (elastase, proteinase 3, and cathepsin G) at 37°C. The T4L/T8L(I380G) mutant was also incubated with 50 nM elastase. Western blotting was performed using an anti-ADAMTS13 metalloprotease domain antibody. The band corresponding to full-length protein (∼180 kDa) is indicated by black arrows.

Article Snippet: Recombinant (r)ADAMTS13 and human ADAMTS13 Quantikine enzyme-linked immunosorbent assay kit (R&D Systems, Minneapolis, MN), kallikrein, FXIIa (Enzyme Research Laboratories, South Bend, IN), thrombin, FXa, FIXa, FXIa, FVIIa, activated protein C (APC), elastase, cathepsin G (Sigma-Aldrich, St Louis, MO), proteinase 3 (Enzo Life Sciences, Inc, Farmingdale, NY), FRETS-VWF73 (AnaSpec, Fremont, CA), AEBSF (4-[2-aminoethyl]benzenesulfonyl fluoride hydrochloride), Sivelestat (Tocris Bioscience, Bristol, United Kingdom), and cathepsin G inhibitor 1 (Cayman Chemical Company, Ann Arbor, MI).

Techniques: Purification, Clinical Proteomics, Mutagenesis, Recombinant, Control, Incubation, Coagulation, Derivative Assay, Western Blot

Journal: Blood Advances

Article Title: Development of a protease-resistant ADAMTS13 to improve stability against proteolytic degradation ∗

doi: 10.1182/bloodadvances.2024015212

Figure Lengend Snippet: Resistance of ADAMTS13 mutants to activated neutrophils. 50 nM WT-, T4L/T8L-, or T4L/T8L(I380G)-ADAMTS13 was incubated with 0 × 10 3 to 50 × 10 3 PMA-activated neutrophils, or 500 × 10 3 neutrophils, as indicated. Reactions took place for 1 hour at 37°C, then samples were removed and separated via SDS-PAGE under reducing conditions. Cleavage was visualized by western blot using an anti-ADAMTS13 metalloprotease domain antibody. Molecular weight references are indicated to the left (kDa). The level of full-length protein (∼180 kDa) is indicated by black arrows.

Article Snippet: Recombinant (r)ADAMTS13 and human ADAMTS13 Quantikine enzyme-linked immunosorbent assay kit (R&D Systems, Minneapolis, MN), kallikrein, FXIIa (Enzyme Research Laboratories, South Bend, IN), thrombin, FXa, FIXa, FXIa, FVIIa, activated protein C (APC), elastase, cathepsin G (Sigma-Aldrich, St Louis, MO), proteinase 3 (Enzo Life Sciences, Inc, Farmingdale, NY), FRETS-VWF73 (AnaSpec, Fremont, CA), AEBSF (4-[2-aminoethyl]benzenesulfonyl fluoride hydrochloride), Sivelestat (Tocris Bioscience, Bristol, United Kingdom), and cathepsin G inhibitor 1 (Cayman Chemical Company, Ann Arbor, MI).

Techniques: Incubation, SDS Page, Western Blot, Molecular Weight

Journal: Blood Advances

Article Title: Development of a protease-resistant ADAMTS13 to improve stability against proteolytic degradation ∗

doi: 10.1182/bloodadvances.2024015212

Figure Lengend Snippet: Activity of ADAMTS13, MDTCS, and T4L/T8L-ADAMTS13 in a microfluidic flow assay. DiOC6-stained platelets were perfused over a microfluidic flow chamber lined with histamine-stimulated HUVECs, forming stable VWF-platelet strings. Channels were then perfused with 5 mL of 5 nM MDTCS, 5 nM WT-ADAMTS13, 5 nM T4L/T8L-ADAMTS13, or ADAMTS13 reaction buffer (n = 3). Using a Leica Stellaris 5 inverted confocal microscope (10× dry lens; λ ex = 490 nm; λ em = 525 nm), videos were captured at 0.77 frames per second; (A) photographic representation of experimental replicate. Experiments occurred at room temperature. The total length of strings within the frame was quantified every 20 frames for 5 frames. Rates were determined using linear regression (B), and differences between rates were assessed using unpaired t tests . Error bars represent standard deviation, and symbols represent 4 experimental conditions: buffer (▲), MDTCS (■), WT-ADAMTS13 (●), T4L/T8L-ADAMTS13 (▼). ∗P < .05, vs MDTCS.

Article Snippet: Recombinant (r)ADAMTS13 and human ADAMTS13 Quantikine enzyme-linked immunosorbent assay kit (R&D Systems, Minneapolis, MN), kallikrein, FXIIa (Enzyme Research Laboratories, South Bend, IN), thrombin, FXa, FIXa, FXIa, FVIIa, activated protein C (APC), elastase, cathepsin G (Sigma-Aldrich, St Louis, MO), proteinase 3 (Enzo Life Sciences, Inc, Farmingdale, NY), FRETS-VWF73 (AnaSpec, Fremont, CA), AEBSF (4-[2-aminoethyl]benzenesulfonyl fluoride hydrochloride), Sivelestat (Tocris Bioscience, Bristol, United Kingdom), and cathepsin G inhibitor 1 (Cayman Chemical Company, Ann Arbor, MI).

Techniques: Activity Assay, Staining, Microscopy, Standard Deviation

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Recombinant human ADAMTS13 attenuates LPS-induced acute kidney injury and renal microangiopathy in murine advanced liver fibrosis by cleaving vWF.

doi: 10.1016/j.bbamcr.2025.120000

Figure Lengend Snippet: Fig. 3. Effect of rhADAMTS13 on plasma ADAMTS13 activity and vWF-Ag balance and coagulation function in AKI-F mice. (A) Experimental design of treatment with rhADAMTS13 against AKI-F mice. (B) Body weight at the end of experiment. (C and D) Plasma ADAMTS13 activity (C) and plasma vWF-Ag level (D). The values are indicated as % NC group. (E) Pearson’s correlation between plasma ADAMTS13 activity and vWF-Ag level in all of experimental mice. (F) Multimer distribution and the ratios of high (H) to low (L) molecular weight of vWF multimers. (G and H) Platelet count and plasma D-dimer level. Data are the mean ± SD, and *: p < 0.05, **: p < 0.01 with significant difference between groups by Student’s t-test (n = 9; B − D, G and H) or Mann−Whitney U test (n = 3; F). NC, normal control without CCl4 and LPS administration; AKI-F, acute kidney injury mice with advanced liver fibrosis.

Article Snippet: Recombinant Human ADAMTS13 (Full Length) Protein, Carrier-free used as rhADAMTS13 was obtained from R&D systems (Minneapolis, MN, USA).

Techniques: Clinical Proteomics, Activity Assay, Coagulation, Molecular Weight, MANN-WHITNEY, Control

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Recombinant human ADAMTS13 attenuates LPS-induced acute kidney injury and renal microangiopathy in murine advanced liver fibrosis by cleaving vWF.

doi: 10.1016/j.bbamcr.2025.120000

Figure Lengend Snippet: Fig. 6. Effect of rADAMTS13 on renal microthrombus and microangiopathy in AKI-F mice. (A) Renal ADAMTS13 (A) and vWF (B) levels. The values are indicated as % NC group. (C) Pearson’s correlation between renal ADAMTS13 and vWF levels in all of experimental mice. (D) Representative photographs of CD41a staining in kidney tissue of NC, D + 2, D + 3 (with and without treatment with rhADAMTS13) groups. (E) Quantification of CD41a+ microthrombus in kidney tissues. DAPI was used as nuclear staining. (F) Microvascular blood flow in the bilateral kidney tissues. (G) Renal mRNA levels of angiogenic factors (Hif1a, Vegfa, Vegfr2, Angpt1, and Tie2). (H) Renal mRNA levels of vascular inflammation markers (Vcam1, Icam1, Sele, and Selp). Gapdh was used as an internal control for qRT-PCR (G and H). The values are indicated as the ratio to NC group (E − H). Data are the mean ± SD, and *: p < 0.05, **: p < 0.01 with significant difference between groups by Student’s t-test (n = 9; A, B, F −H, n = 6; E). NC, normal control without CCl4 and LPS administration; AKI-F, acute kidney injury mice with advanced liver fibrosis; rAD, recombinant ADAMTS13.

Article Snippet: Recombinant Human ADAMTS13 (Full Length) Protein, Carrier-free used as rhADAMTS13 was obtained from R&D systems (Minneapolis, MN, USA).

Techniques: Staining, Control, Quantitative RT-PCR, Recombinant