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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. 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.In non-model organisms, evolutionary questions are frequently addressed using reduced representation sequencing techniques due to their low cost, ease of use, and because they do not require genomic resources such as a reference genome. However, evidence is accumulating that such techniques may be affected by specific biases, questioning the accuracy of obtained genotypes, and as a consequence, their usefulness in evolutionary studies. Here we introduce three strategies to estimate genotyping error rates from such data through the comparison to high quality genotypes obtained with a different technique, from individual replicates, or from a population sample when assuming Hardy-Weinberg equilibrium. Applying these strategies to data obtained with Restriction site Associated DNA sequencing (RAD-seq), arguably the most popular reduced representation sequencing technique, revealed per-allele genotyping error rates that were much higher than sequencing error rates, particularly at heterozygous sites that were wrongly inferred as homozygous. As we exemplify through the inference of genome-wide and local ancestry of well characterized hybrids of two Eurasian poplar (Populus) species, such high error rates may lead to wrong biological conclusions. By properly accounting for these error rates in downstream analyses, either by incorporating genotyping errors directly or by recalibrating genotype likelihoods, we were nevertheless able to use the RAD-seq data to support biologically meaningful and robust inferences of ancestry among Populus hybrids. Based on these findings, we strongly recommend carefully assessing genotyping error rates in reduced representation sequencing experiments, and to properly account for these in downstream analyses, for instance using the tools presented here. This article is protected by copyright. All rights reserved.In recent years, tools for functional genomic studies have become increasingly feasible for use by evolutionary anthropologists. In this review, we provide brief overviews of several exciting in vitro techniques that can be paired with "-omics" approaches (e.g., genomics, epigenomics, transcriptomics, proteomics, and metabolomics) for potentially powerful evolutionary insights. These in vitro techniques include ancestral protein resurrection, cell line experiments using primary, immortalized, and induced pluripotent stem cells, and CRISPR-Cas9 genetic manipulation. We also discuss how several of these methods can be used in vivo, for transgenic organism studies of human and nonhuman primate evolution. Throughout this review, we highlight example studies in which these approaches have already been used to inform our understanding of the evolutionary biology of modern and archaic humans and other primates while simultaneously identifying future opportunities for anthropologists to use this toolkit to help answer additional outstanding questions in evolutionary anthropology. © 2020 Wiley Periodicals, Inc.OBJECTIVE The aim of the current research was to identify the extent to which reward sensitivity and impulsivity were related to food addiction. METHOD Forty-five studies, published from 2009 to June 2019, investigating reward sensitivity and/or impulsivity with food addiction as measured by the Yale Food Addiction Scale were reviewed. RESULTS Reward sensitivity, as measured by the Sensitivity to Reward (SR) scale, was positively associated with food addiction in two studies, but failed to yield consistent results in other studies when measured with the Behavioral Inhibition/Behavioral Activation Scales. Self-report impulsivity, as measured by the Barratt Impulsiveness Scale (BIS-11), was consistently associated with food addiction, with attentional impulsivity and motor impulsivity the most consistent subscales. Similarly, food addiction was also consistently associated with Negative Urgency, Positive Urgency, and Lack of Perseverance as measured by the UPPS-P Impulsive Behavior Scale. Food addiction was inconsistently associated with disinhibition, as measured by behavioral tasks, indicating food addiction appears more aligned with self-report measures of impulsivity. CONCLUSIONS Research in this field is dominated by university student, overweight and obese samples. Additional research is required to further tease out these relationships. © 2020 John Wiley & Sons, Ltd and Eating Disorders Association.PURPOSE The present study sought to design a multi-functional fusion peptide with hydroxyapatite (HA) binding domain (HABD) and heparin binding domain (HBD). METHODS The 74 amino acid fusion peptide contained N-terminus of the fibrinogen β chain (β 15-66), double G4S-linker and 12 residues with HA affinity. This construct was designed, synthesized and cloned into pET21a(+) vector and expressed in E. coli. 2',3'-cGAMP RESULTS HABD facilitated purification of the fusion peptide by HA affinity chromatography. Kinetic peptide binding and release on HA scaffold showed sustained release of peptide for up to 16 days. Competitive ELISA and intrinsic fluorescence assays were applied to determine HBD affinity to bone morphogenetic protein-2 (BMP-2). The disassociation rate constant (Kd ) for HBD and rhBMP-2 was approximately 9.2-12 nM. CONCLUSION The fusion peptide developed in the present study, allowed for streamlined purification on HA affinity chromatography, as well as sustained release from HA scaffold, attributed to its HABD.