The calculated Fukui function inferred the local softness and electrophilicity indices of used solute compounds. © 2020 John Wiley & Sons, Ltd.Native mass spectrometry is an emerging technique in biology that gives the possibility to study non-covalently bound complexes with high sensitivity and accuracy. It thus allows the characterization of macromolecular assemblies, assessing their mass and stoichiometries and mapping the interacting surfaces. In this review, we discuss the application of native mass spectrometry to dynamic molecular machines based on multiple weak interactions. In the study of these machines, it is crucial to understand which and under which conditions various complexes form at any time point. We focus on the specific example of the iron-sulfur cluster biogenesis machine because this is an archetype of a dynamic machine that requires very specific and demanding experimental conditions, such as anaerobicity and the need of retaining the fold of marginally folded proteins. We describe the advantages, challenges and current limitations of the technique by providing examples from our own experience and suggesting possible future solutions. This article is protected by copyright. All rights reserved.Follicular helper T (TFH) cell provides germinal centre (GC) B cell with critical signals for autoantibody production in the immunopathogenesis and progression of autoimmune hepatitis (AIH). However, the immunoregulatory functions of follicular regulatory T (TFR) cell in AIH are still unclear. The numbers of circulating TFR/TFH cells were measured in AIH patients. Moreover, we established experimental autoimmune hepatitis (EAH) model to examine the function of TFR cells on B-cell differentiation and autoantibody production in vivo and vitro. AIH patients had significantly increased numbers of TFH cells and decreased numbers of TFR cells as well as imbalanced TFR/TFH-type cytokines (IL-10, TGF-β1 and IL-21) compared with healthy controls (HCs). In addition, TFR cell numbers negatively correlated with TFH cell numbers. Also, serum hypergammaglobulinaemia (IgG and IgM) concentration negatively correlated the levels of serum IL-21, but positively correlated with the levels of serum IL-10 in AIH patients. Furthermore, in comparison with control group, significantly higher frequencies of spleen TFR cells but lower frequencies of spleen TFH cells were detected in the EAH group. Further analysis found that TFR cells simultaneously express the phenotypic characteristics of Treg and TFH cells, but exercise as negative regulators of autoantibody production in vitro culture. Our findings demonstrated that dysregulated between TFR and TFH cells might cause excessive production of autoantibodies and destruction of the immune homeostasis, leading to the immunopathological process in AIH. © 2020 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd.INTRODUCTION Many recombinant and modified FIX products have been, and continue to be, developed with the aim of improving treatment for patients with haemophilia B. One such new product is dalcinonacog alfa, a recombinant FIX with modifications to provide improved features such as subcutaneous administration. AIM In view of previously observed assay discrepancies with modified FIX therapeutics, the aim of this study was to assess potential discrepancies in potency measurement of dalcinonacog alfa between and within different assay methods. METHODS Potency of dalcinonacog alfa was measured against the 5th International Standard (IS) for FIX Concentrate and the 4th IS for FIX Plasma by One-Stage Clotting Assay, using 9 different APTT reagents and 2 commercially available FIX chromogenic kits. Plasma-derived concentrate and recombinant FIX samples were also included for comparison in every assay. RESULTS Substantial discrepancies were observed when assaying dalcinonacog alfa using the one-stage clotting assay against both standards. No statistically valid results were obtained when testing dalcinonacog alfa using either chromogenic kit. Increasing the incubation time with the activation reagent in both chromogenic kits resulted in valid assays and increased the potency to become more in line with potencies by one-stage clotting assays. Increasing the incubation time in the chromogenic kits had no effect on the potencies of the plasma-derived or recombinant samples. However, incubation time influenced in the one-stage clotting assay using Dapttin. CONCLUSIONS Within and between assay method discrepancy was found when assaying dalcinonacog alfa. Methods for potency labelling and clinical monitoring should be given careful consideration. © 2020 The Authors. Haemophilia published by John Wiley & Sons Ltd.INTRODUCTION Obstructive sleep apnea (OSA) is an oxidative stress disease, which has been considered to be a notable risk and associated with increased cardiovascular morbidity and mortality. Thiol-disulfide homeostasis is as a novel indicator of oxidative stress. OBJECTIVES We aimed to evaluate thiol-disulfide homeostasis in a large patient population with OSA. METHODS A total of 230 with OSA and 40 healthy controls were included in the study. Inclusion criteria for OSA patients are having apnoea-hypopnoea index of ≥5/hour, being more than 18 years of age and having no previous treatment for OSA. Thiol-disulfide analysis was done for the patients and control group. Blood thiol-disulfide homeostasis was analysed using the new automatic method, developed by Erel and Neşelioğlu. RESULTS Among all OSA subjects, 149 (64.8%) were males and the mean ages of the patients were 53.38 ± 10.22.  https://www.selleckchem.com/btk.html  Total thiol, native thiol (SH) and disulfide (SS) levels were significantly lower in OSA group compared to the control group (P  less then  .001, P  less then  .001 and P = .039 respectively). Also, total thiol and native thiol (SH) were significantly different between the groups according to OSA severity (mild-moderate to severe OSA) (P  less then  .001 and P  less then  .001 respectively). Thiol-disulfide redox parameters were correlated with apnoea-hypopnoea index (AHI) scores. CONCLUSION The present prospective study showed that thiol/disulfide homeostasis was unbalanced in OSA patients. Especially, in OSA patients have low level of thiol/disulfide redox parameters when compared to healthy subjects. Evaluating thiol-disulfide homeostasis in OSA may be a contributing aspect to assessment and monitoring of the patient. © 2020 John Wiley & Sons Ltd.