Table_1_Effects of physical activity and sedentary behaviors on cardiovascular disease and the risk of all-cause mortality in overweight or obese middle-aged and older adults.docx

AimThe aim of this study was to respectively explore the relationships between physical activity and sedentary behaviors and cardiovascular disease (CVD) and all-cause mortality risk in overweight/obese middle-aged and older patients, and also assess the interaction between physical activity and sedentary behaviors.MethodsData of middle-aged and older adults with body mass index (BMI) ≥25 kg/m2 were extracted from the National Health and Nutrition Examination Surveys (NHANES) database in 2007–2018 in this retrospective cohort study. Weighted univariate and multivariate logistic regression analyses were used to explore the associations between physical activity and sedentary behaviors and CVDs; weighted univariate and multivariate Cox regression analyses were used to explore the relationships between physical activity and sedentary behaviors with the risk of all-cause mortality. The interaction effect between physical activity and sedentary behaviors on CVD and all-cause mortality was also assessed. We further explored this interaction effect in subgroups of age and BMI. The evaluation indexes were odds ratios (ORs), hazard ratios (HRs), and 95% confidence intervals (CIs).ResultsAmong 13,699 eligible patients, 1,947 had CVD, and 1,560 died from all-cause mortality. After adjusting for covariates, patients who had high sedentary time seemed to have both high odds of CVD [OR = 1.24, 95% CI: (1.06–1.44)] and a high risk of all-cause mortality [HR = 1.20, 95% CI: (1.06–1.37)]. Furthermore, being insufficiently active was linked to high odds of CVD [OR = 1.24, 95% CI: (1.05–1.46)] as well as a high risk of all-cause mortality [HR = 1.32, 95% CI: (1.15–1.51)]. High sedentary time and being insufficiently active had an interaction effect on both high odds of CVD [OR = 1.44, 95% CI: (1.20–1.73)] and high risk of all-cause mortality [HR = 1.48, 95% CI: (1.24–1.76)]. Individuals of different ages with/without obesity need to focus on the potential CVD/mortality risk of high sedentary time and low physical activity (all P ConclusionReducing sedentary time combined with increasing physical activity may benefit health by reducing both the risk of CVD and all-cause mortality in overweight or obese middle-aged and older adults.</p

Discovery of Selective Small-Molecule Inhibitors for the β‑Catenin/T-Cell Factor Protein–Protein Interaction through the Optimization of the Acyl Hydrazone Moiety

Acyl hydrazone is an important functional group for the discovery of bioactive small molecules. This functional group is also recognized as a pan assay interference structure. In this study, a new small-molecule inhibitor for the β-catenin/Tcf protein–protein interaction (PPI), ZINC02092166, was identified through AlphaScreen and FP assays. This compound contains an acyl hydrazone group and exhibits higher inhibitory activities in cell-based assays than biochemical assays. Inhibitor optimization resulted in chemically stable derivatives that disrupt the β-catenin/Tcf PPI. The binding mode of new inhibitors was characterized by site-directed mutagenesis and structure–activity relationship studies. This series of inhibitors with a new scaffold exhibits dual selectivity for β-catenin/Tcf over β-catenin/cadherin and β-catenin/APC PPIs. One derivative of this series suppresses canonical Wnt signaling, downregulates the expression of Wnt target genes, and inhibits the growth of cancer cells. This compound represents a solid starting point for the development of potent and selective β-catenin/Tcf inhibitors

Structure-Based Optimization of Small-Molecule Inhibitors for the β‑Catenin/B-Cell Lymphoma 9 Protein–Protein Interaction

Structure-based optimization was conducted to improve the potency, selectivity, and cell-based activities of β-catenin/B-cell lymphoma 9 (BCL9) inhibitors based on the 4′-fluoro-<i>N</i>-phenyl-[1,1′-biphenyl]-3-carboxamide scaffold, which was designed to mimic the side chains of the hydrophobic α-helical hot spots at positions <i>i</i>, <i>i</i> + 3, and <i>i</i> + 7. Compound <b>29</b> was found to disrupt the β-catenin/BCL9 protein–protein interaction (PPI) with a <i>K</i>_i of 0.47 μM and >1900-fold selectivity for β-catenin/BCL9 over β-catenin/E-cadherin PPIs. The proposed binding mode of new inhibitors was consistent with the results of site-directed mutagenesis and structure–activity relationship studies. Cell-based studies indicated that <b>29</b> disrupted the β-catenin/BCL9 interaction without affecting the β-catenin/E-cadherin interaction, selectively suppressed transactivation of Wnt/β-catenin signaling, downregulated expression of Wnt target genes, and inhibited viability of Wnt/β-catenin-dependent cancer cells in dose-dependent manners. A comparison of the biochemical and cell-based assay results offered the directions for future inhibitor optimization

Structure-Based Optimization of Small-Molecule Inhibitors for the β‑Catenin/B-Cell Lymphoma 9 Protein–Protein Interaction

Structure-based optimization was conducted to improve the potency, selectivity, and cell-based activities of β-catenin/B-cell lymphoma 9 (BCL9) inhibitors based on the 4′-fluoro-<i>N</i>-phenyl-[1,1′-biphenyl]-3-carboxamide scaffold, which was designed to mimic the side chains of the hydrophobic α-helical hot spots at positions <i>i</i>, <i>i</i> + 3, and <i>i</i> + 7. Compound <b>29</b> was found to disrupt the β-catenin/BCL9 protein–protein interaction (PPI) with a <i>K</i>_i of 0.47 μM and >1900-fold selectivity for β-catenin/BCL9 over β-catenin/E-cadherin PPIs. The proposed binding mode of new inhibitors was consistent with the results of site-directed mutagenesis and structure–activity relationship studies. Cell-based studies indicated that <b>29</b> disrupted the β-catenin/BCL9 interaction without affecting the β-catenin/E-cadherin interaction, selectively suppressed transactivation of Wnt/β-catenin signaling, downregulated expression of Wnt target genes, and inhibited viability of Wnt/β-catenin-dependent cancer cells in dose-dependent manners. A comparison of the biochemical and cell-based assay results offered the directions for future inhibitor optimization

GO categories of biological process, cellular component and molecular function for the transcriptome of the <i>A</i>. <i>sparsifolia</i> primary roots.

<p>The right y-axis shows the number of genes in a category, and the left y-axis indicates the percentage of a specific category of genes in that main category.</p

Effect of water stress induced by PEG-6000 on development of the <i>Alhagi sparsifolia</i> primary roots.

<p>(A) Seeds of <i>A</i>. <i>sparsifolia</i> (Scale bar = 2.0 cm). (B) Typical phenotype of the <i>A</i>. <i>sparsifolia</i> seedlings at 7 days post treatment with PEG-6000 (Scale bar = 1.0 cm). The <i>A</i>. <i>sparsifolia</i> seeds were first treated with concentrated sulfuric acid (98%) for 20 min, and then put on fully wetted filter paper at 25°C in the dark for 24 h. The germinated seeds were selected and transferred to petri dishes containing filter paper saturated with the different percentages of PEG-6000 solution (0, 90, 150, 220, 270 and 320 g/L), and were remained at 25°C in the dark for 3 days followed by 4 additional days but under 16-h light/8-h dark. At least 15 seedlings were included in each different treatment, and three independent experiments were preformed. (C) Average length of the primary roots of <i>A</i>. <i>sparsifolia</i> treated with PEG-6000 at 7 days. Error bars represent SE. P values determined by Student t test (** p < 0.01).</p

Summary of unigene annotations against public databases.

<p>Abbreviations: NR, non-redundant protein sequence; NT, non-redundant nucleotide; Pfam, Protein Family Database; KEGG, Kyoto Encyclopedia of Genes and Genomes database; KOG, euKaryotic Ortholog Groups; GO, Gene Ontology.</p><p>Summary of unigene annotations against public databases.</p

GO categories of biological process (BP), cellular component (CC) and molecular function (MF) for the early water-stress responding genes (P-6h <i>vs</i>. P-0h) in <i>A</i>. <i>sparsifolia</i> primary roots.

<p>(A) the early water-stress inducible genes; (B) the early water-stress repressed genes. The right y-axis shows the number of genes in a category, and the left y-axis indicates the percentage of a specific category of genes in that main category.</p

Rational Design of Selective Small-Molecule Inhibitors for β‑Catenin/B-Cell Lymphoma 9 Protein–Protein Interactions

Selective inhibition of α-helix-mediated protein–protein interactions (PPIs) with small organic molecules provides great potential for the discovery of chemical probes and therapeutic agents. Protein Data Bank data mining using the HippDB database indicated that (1) the side chains of hydrophobic projecting hot spots at positions <i>i</i>, <i>i</i> + 3, and <i>i</i> + 7 of an α-helix had few orientations when interacting with the second protein and (2) the hot spot pockets of PPI complexes had different sizes, shapes, and chemical groups when interacting with the same hydrophobic projecting hot spots of α-helix. On the basis of these observations, a small organic molecule, 4′-fluoro-<i>N</i>-phenyl-[1,1′-biphenyl]-3-carboxamide, was designed as a generic scaffold that itself directly mimics the binding mode of the side chains of hydrophobic projecting hot spots at positions <i>i</i>, <i>i</i> + 3, and <i>i</i> + 7 of an α-helix. Convenient decoration of this generic scaffold led to the selective disruption of α-helix-mediated PPIs. A series of small-molecule inhibitors selective for β-catenin/B-cell lymphoma 9 (BCL9) over β-catenin/cadherin PPIs was designed and synthesized. The binding mode of new inhibitors was characterized by site-directed mutagenesis and structure–activity relationship studies. This new class of inhibitors can selectively disrupt β-catenin/BCL9 over β-catenin/cadherin PPIs, suppress the transactivation of canonical Wnt signaling, downregulate the expression of Wnt target genes, and inhibit the growth of Wnt/β-catenin-dependent cancer cells