Transient-state and steady-state kinetics of the oxidation of aliphatic and aromatic thiols by horseradish peroxidase

AbstractIn the course of oxidation of thiols by peroxidases thiyl radicals are formed which are known to undergo several free-radical conjugative reactions, among others leading to hydrogen peroxide formation. The present paper for the first time presents a comparative transient-state and steady-state investigation of the reaction of 15 aliphatic and aromatic mono- and dithiols with horseradish peroxidase (HRP). Both sequential-stopped-flow spectrophotometric investigations of the reaction of HRP intermediates Compound I (k2) and Compound II (k3) with thiols and measurements of the overall thiol oxidation and the simultaneous oxygen consumption in the presence and absence of exogenously added hydrogen peroxide (10 μM) have been performed. With HRP as thiyl radical generator it was shown that three groups of thiols have to be distinguished: (i) Aromatic thiols (e.g. thiophenol, 2-mercaptopurine) were excellent electron donors of both Compounds (k2: 104–107 M−1 s−1 and k3: 103–106 M−1 s−1); however, the overall reaction was shown to depend on addition of hydrogen peroxide, indicating insufficient peroxide regeneration by arylthiyl radicals. (ii) Aliphatic thiols which were extremely bad substrates (k3 <10 M−1 s−1) for HRP (e.g. homocysteine, glutathione) and/or have a pKa,SH >9.5 (e.g. N-acetylcysteine, α-lipoic acid) were also shown to depend on exogenously added H2O2 to maintain the peroxidasic reaction, whereas (iii) with those thiols with rates of k3 between 11 and 1600 M−1 s−1 (e.g. cysteine, cysteamine, cysteine methyl ester, cysteine ethyl ester) and/or with a pKa,SH <8 (penicillamine) thiol oxidation was independent of exogenously added hydrogen peroxide, indicating sufficient hydrogen peroxide regeneration

Fabric-mechanical property relationships of trabecular bone allografts are altered by supercritical CO2 treatment and gamma sterilization

Tissue grafts are implanted in orthopedic surgery every day. In order to minimize infection risk, bone allografts are often delipidated with supercritical CO2 and sterilized prior to implantation. This treatment may, however, impair the mechanical behavior of the bone graft tissue. The goal of this study was to determine clinically relevant mechanical properties of treated/sterilized human trabecular bone grafts, e.g. the apparent modulus, strength, and the ability to absorb energy during compaction. They were compared with results of identical experiments performed previously on untreated/fresh frozen human trabecular bone from the same anatomical site (Charlebois, 2008). We tested the hypothesis that the morphology–mechanical property relationships of treated cancellous allografts are similar to those of fresh untreated bone. The morphology of the allografts was determined by μCT. Subsequently, cylindrical samples were tested in unconfined and confined compression. To account for various morphologies, the experimental data was fitted to phenomenological mechanical models for elasticity, strength, and dissipated energy density based on bone volume fraction (BV/TV) and the fabric tensor determined by MIL. The treatment/sterilization process does not appear to influence bone graft stiffness. However, strength and energy dissipation of the bone grafts were found to be significantly reduced by 36% to 47% and 66% to 81%, respectively, for a broad range of volume fraction (0.14 < BV/TV < 0.39) and degree of anisotropy (1.24 < DA < 2.18). Since the latter properties are strongly dominated by BV/TV, the clinical consequences of this reduction can be compensated by using grafts with lower porosity. The data of this study suggests that an increase of 5–10% in BV/TV is sufficient to compensate for the reduced post-yield mechanical properties of treated/sterilized bone in monotonic compression. In applications where graft stiffness needs to be matched and strength is not a concern, treated allograft with the same BV/TV as an appropriate fresh bone graft may be used

The two-Cys peroxiredoxin Bas1: Insight in a new family of plant peroxidases

Baier M, Dietz K-J. The two-Cys peroxiredoxin Bas1: Insight in a new family of plant peroxidases. In: Obinger C, Burner U, Ebermann R, Penel C, Greppin H, eds. Plant peroxidases, biochemistry and physiology. Genève: Laboratoire de Biochimie et Physiologie Vègètales, Univ. de Genève; 1996: 204-209

Immediate ROI Search for 3-D Medical Images

The objective of this work is a scalable, real-time, visual search engine for 3-D medical images, where a user is able to select a query Region Of Interest (ROI) and automatically detect the corresponding regions within all returned images. We make three contributions: (i) we show that with appropriate off-line processing, images can be retrieved and ROIs registered in real time; (ii) we propose and evaluate a number of scalable exemplar-based image registration schemes; (iii) we propose a discriminative method for learning to rank the returned images based on the content of the ROI. The retrieval system is demonstrated on MRI data from the ADNI dataset, and it is shown that the learnt ranking function outperforms the baseline. © 2013 Springer-Verlag