Renal Allograft Dysfunction: Evaluation with Shear-wave Sonoelastography.

Purpose To evaluate whether shear-wave sonoelastography can help differentiate stable renal allograft from acute allograft dysfunction and chronic allograft dysfunction and to correlate shear-wave sonoelastography measurements with resistive index (RI), serum creatinine level, estimated glomerular filtration rate (eGFR) obtained with the Nankivell equation, and biopsy findings. Materials and Methods A prospective study of 60 patients who had undergone renal transplantation was conducted between October 2014 and March 2016. Patients were classified as having stable allograft, acute allograft dysfunction, or chronic allograft dysfunction on the basis of clinical parameters. Mean parenchymal stiffness was compared. The Banff score was used wherever applicable. Receiver operating characteristic curves were drawn to evaluate the feasibility of differentiation. Results Thirty patients had graft dysfunction (acute in 19 patients and chronic in 11). Mean parenchymal stiffness values in stable allograft, acute allograft dysfunction, and chronic allograft dysfunction were 8.51 kPa ± 2.44, 11.06 kPa ± 2.91, and 24.50 kPa ± 4.49, respectively (stable vs acute dysfunction, P = .010; stable vs chronic dysfunction, P < .001; acute sysfunction vs chronic dysfunction, P < .001). The allograft parenchymal stiffness values for patients with Banff grade I (mild interstitial fibrosis and tubular atrophy) differed significantly from those with Banff grade II (moderate interstitial fibrosis and tubular atrophy) (P = .02). Parenchymal stiffness showed a negative correlation with eGFR (r = -0.725; P < .001) and a positive correlation with RI (r = 0.562; P < .001) and serum creatinine level (r = 0.714; P < .001). The sensitivity was 73.68% and specificity was 80% in the differentiation of stable graft from acute graft dysfunction (threshold value, 10.11 kPa). Conclusion Shear-wave sonoelastographic evaluation of renal parenchymal stiffness may help differentiate stable allograft from acute and chronic allograft dysfunction. The inverse correlation of parenchymal stiffness with eGFR and positive correlation with RI and serum creatinine level show that shear-wave sonoelastography may reflect functional status of the renal allograft.

Multiple metalloproteinases process protransforming growth factor-alpha (proTGF-alpha).

Shedding of TNF-alpha requires a single cleavage event, whereas the ectodomain of proTGF-alpha is cleaved at N-proximal (N-terminal) and membrane proximal (C-terminal) sites to release mature TGF-alpha. Tumor necrosis factor-alpha converting enzyme (TACE) was shown to have a central role in the shedding of both factors. Here we show that cleavage of the proTGF-alpha C-terminal site, required for release of mature growth factor, is less sensitive to a panel of hydroxamates than TNF-alpha processing. Recombinant TACE cleaves TNF-alpha and N-terminal TGF-alpha peptides 50-fold more efficiently than the C-terminal TGF-alpha peptide. Moreover, fractionation of rat liver epithelial cell membranes yields two populations: one contains TACE and cleaves peptides corresponding to TNF-alpha and both proTGF-alpha processing sites, while the other lacks detectable TACE and cleaves only the C-terminal proTGF-alpha processing site. Activities in both fractions are inhibited by hydroxamates and EDTA but not by cysteine, aspartate, or serine protease inhibitors. Both membrane fractions also contain ADAM 10. ADAM 10 correctly cleaves peptides and a soluble form of precursor TGF-alpha (proTGFecto) at the N-terminal site but not the C-terminal site. However, the kinetics of N-terminal peptide cleavage by ADAM 10 are 90-fold less efficient than TACE. Our findings indicate that while TACE is an efficient proTGF-alpha N-terminal convertase, a different activity, distinguishable from TACE, exists that can process proTGF-alpha at the C-terminal site. A model that accounts for these findings and the requirement for TACE in TGF-alpha shedding is proposed.

The tumor necrosis factor-alpha converting enzyme (TACE): a unique metalloproteinase with highly defined substrate selectivity.

TNF alpha converting enzyme (TACE) processes precursor TNF alpha between Ala76 and Val77, yielding a correctly processed bioactive 17 kDa protein. Genetic evidence indicates that TACE may also be involved in the shedding of other ectodomains. Here we show that native and recombinant forms of TACE efficiently processed a synthetic substrate corresponding to the TNF alpha cleavage site only. For all other substrates, conversion occurred only at high enzyme concentrations and prolonged reaction times. Often, cleavage under those conditions was accompanied by nonspecific reactions. We also compared TNF alpha cleavage by TACE to cleavage by those members of the matrix metalloproteinase (MMP) family previously implied in TNF alpha release. The specificity constants for TNF alpha cleavage by the MMPs were approximately 100-1000-fold slower relative to TACE. MMP 7 also processed precursor TNF alpha at the correct cleavage site but did so with a 30-fold lower specificity constant relative to TACE. In contrast, MMP 1 processed precursor TNF alpha between Ala74 and Gln75, in addition to between Ala76 and Val77, while MMP 9 cleaved this natural substrate solely between Ala74 and Gln75. Additionally, the MMP substrate Dnp-PChaGC(Me)HK(NMA)-NH(2) was not cleaved at all by TACE, while collagenase (MMP 1), gelatinase (MMP 9), stromelysin 1 (MMP 3), and matrilysin (MMP 7) all processed this substrate efficiently. All of these results indicate that TACE is unique in terms of its specificity requirements for substrate cleavage.