The Distinct Expressions of Integrins αDβ2 and αMβ2 Differently Regulate Macrophage Migration in 3D Matrix in vitro and in Tissue during Inflammation

Chronic inflammation is an essential mechanism during the development of cardiovascular and metabolic diseases. The outcome of diseases depends on the balance between the migration and accumulation of macrophages in damaged tissues. Macrophage motility is highly regulated by adhesive receptors, integrins. Namely, intermediate expression of integrin supports macrophage migration, while a high integrin density inhibits it. Our studies are focused on evaluation of the contribution of related integrins αDβ2 and αMβ2 to macrophage migration and development of chronic inflammation. We found that integrin αDβ2 is upregulated on M1-macrophages in vitro and pro-inflammatory macrophages in atherosclerotic lesions. Interestingly, the expression of ligand-sharing integrin αMβ2 remains unaltered. Using in vitro three-dimensional migration and in vivo tracking of adoptively-transferred fluorescently-labeled macrophages during the resolution of inflammation, we found that robust adhesion of M1-activated macrophages translates to weak 3D migration, which depends on the high expression of αDβ2, since αD-deficiency decreases M1-macrophage adhesion and improves macrophage migration. In contrast, αD- and αM-knockouts decrease M2-macrophages migration, demonstrating that moderate integrin expression supports cell motility. In model of high fat diet-induced diabetes, αD-deficiency prevents the retention of inflammatory macrophages in adipose tissue and improves metabolic parameters, while αM-deficiency does not affect macrophage accumulation. We detected a new ligand for integrins αMβ2 and αDβ2, 2-(ω-carboxyethyl)pyrrole (CEP). CEP is preferentially generated during inflammation-mediated oxidation and forms adduct with ECM proteins generating novel substrate for αMβ2 and αDβ2. Targeting CEP-dependent macrophage adhesion can be a useful approach to control αDβ2-mediated chronic inflammation. Using specially designed peptide library, protein-protein interaction and adhesion assay, we identified a peptide, called P5, which significantly inhibited αD-CEP binding. P5 peptide regulates macrophage migration in three-dimensional matrix in vitro and reduced macrophage accumulation during thioglycollate-induced peritoneal inflammation. Effect of P5 is completely eliminated in αD-deficient macrophages. Tracking of adoptively-transferred fluorescently-labeled WT and αD-/- monocytes in diabetic mice confirmed that αD-dependent inhibition of macrophage accumulation in adipose tissue is mediated by P5 peptide. Taken together, these results demonstrate the importance of αDβ2 and αDβ2-CEP interaction for the accumulation of infiltrating macrophages during inflammation and propose P5 peptide as a potential inhibitor of atherogenesis and diabetes

Kinesio taping for ankle sprain in youth athlete:a protocol for systematic review and meta-analysis

Background: Ankle sprain is considered a major problem that may hinder youth athletes' athletic development because it will lead to ongoing dysfunction, reoccurrence of ankle sprain, chronic ankle instability, and posttraumatic osteoarthritis. Kinesio taping (KT) is a therapeutic taping technique that has been widely used in the treatment of various ankle issues including sprained ankles and in the prevention of ankle sprains. It can not only provide the injured ankle with support during the rehabilitation phase, but also enhance the ankle stability during activity. However, the available evidence regarding its effectiveness in the treatment and prevention of ankle sprain is inconsistent. Therefore, a systematic review will help clinicians and coaches better understand the application of KT in clinical and training practices. This study is to systematically review the literature on the use of KT to improve outcomes including ankle function, proprioception, and pain and to evaluate the effectiveness of KT in the treatment and prevention of ankle sprain injuries. Methods: A comprehensive electronic search of the literature will be undertaken in the following databases: PubMed, CINAHL, SPORTDiscus, Cochrane library, Web of Science and Scopus from 1979 to August 2022. The Physiotherapy Evidence Database scale will be used to assess the methodological quality of all included studies and RevMan 5.3 (Copenhagen, The Nordic Cochrane Centre) for the data analysis. Results: This study will provide a standardized evaluation and comparison for effects of KT on the treatment and prevention of ankle sprains in youth athletes. Conclusion: This review will provide the evidence of the effectiveness of KT used in the treatment and prevention of ankle sprain in youth athletes. This review will also provide directions and recommendations for future research and clinical practices targeting treatment and prevention of ankle sprains in youth athletes.</p

Distinct Migratory Properties of M1, M2, and Resident Macrophages Are Regulated by α\u3csub\u3ed\u3c/sub\u3eβ\u3csub\u3e2\u3c/sub\u3eand α\u3csub\u3em\u3c/sub\u3eβ\u3csub\u3e2\u3c/sub\u3eIntegrin-Mediated Adhesion

Chronic inflammation is essential mechanism during the development of cardiovascular and metabolic diseases. The outcome of diseases depends on the balance between the migration/accumulation of pro-inflammatory (M1) and anti-inflammatory (M2) macrophages in damaged tissue. The mechanism of macrophage migration and subsequent accumulation is still not fully understood. Currently, the amoeboid adhesion-independent motility is considered essential for leukocyte migration in the three-dimensional environment. We challenge this hypothesis by studying the contribution of leukocyte adhesive receptors, integrins αMβ2, and αDβ2, to three-dimensional migration of M1-polarized, M2-polarized, and resident macrophages. Both integrins have a moderate expression on M2 macrophages, while αDβ2 is upregulated on M1 and αMβ2 demonstrates high expression on resident macrophages. The level of integrin expression determines its contribution to macrophage migration. Namely, intermediate expression supports macrophage migration, while a high integrin density inhibits it. Using in vitro three-dimensional migration and in vivo tracking of adoptively-transferred fluorescently-labeled macrophages during the resolution of inflammation, we found that strong adhesion of M1-activated macrophages translates to weak 3D migration, while moderate adhesion of M2-activated macrophages generates dynamic motility. Reduced migration of M1 macrophages depends on the high expression of αDβ2, since αD-deficiency decreased M1 macrophage adhesion and improved migration in fibrin matrix and peritoneal tissue. Similarly, the high expression of αMβ2 on resident macrophages prevents their amoeboid migration, which is markedly increased in αM-deficient macrophages. In contrast, αD- and αM-knockouts decrease the migration of M2 macrophages, demonstrating that moderate integrin expression supports cell motility. The results were confirmed in a diet-induced diabetes model. αD deficiency prevents the retention of inflammatory macrophages in adipose tissue and improves metabolic parameters, while αM deficiency does not affect macrophage accumulation. Summarizing, β2 integrin-mediated adhesion may inhibit amoeboid and mesenchymal macrophage migration or support mesenchymal migration in tissue, and, therefore, represents an important target to control inflammation

Inhibition of Integrin α\u3csub\u3ed\u3c/sub\u3eβ\u3csub\u3e2\u3c/sub\u3e-Mediated Macrophage Adhesion to End Product of Docosahexaenoic Acid (DHA) Oxidation Prevents Macrophage Accumulation During Inflammation

A critical step in the development of chronic inflammatory diseases is the accumulation of proinflammatory macrophages in the extracellular matrix (ECM) of peripheral tissues. The adhesion receptor integrin αDβ2 promotes the development of atherosclerosis and diabetes by supporting macrophage retention in inflamed tissue. We recently found that the end product of docosahexaenoic acid (DHA) oxidation, 2-(ω-carboxyethyl)- pyrrole (CEP), serves as a ligand forαDβ2.CEPadduct withECM is generated during inflammation-mediated lipid peroxidation. The goal of this project was to identify a specific inhibitor for αDβ2-CEP interaction that can prevent macrophage accumulation. Using a specially designed peptide library, Biacore-detected protein-protein interaction, and adhesion of integrin-transfected HEK 293 cells, we identified a sequence (called P5 peptide) that significantly and specifically inhibited αD-CEP binding. In the model of thioglycollate-induced peritoneal inflammation, the injection of cyclic P5 peptide reduced 3-fold the macrophage accumulation in WT mice but had no effect in αD-deficient mice. The tracking of adoptively transferred, fluorescently labeled WT and αD-/- monocytes in the model of peritoneal inflammation and in vitro two-dimensional and three-dimensional migration assays demonstrated thatP5peptide does not affectmonocytetransendothelial migration or macrophage efflux from the peritoneal cavity but regulates macrophage migration through the ECM. Moreover, the injection ofP5peptide intoWTmiceona high-fat diet prevents macrophage accumulation in adipose tissue in anαDβ2-dependent manner.Takentogether, these resultsdemonstratetheimportance of αDβ2-mediated macrophage adhesion for the accumulation of infiltrating macrophages in the inflamed ECM and propose P5 peptide as a potential inhibitor of atherogenesis and diabetes

Computable features required to evaluate the efficacy of drugs and a universal algorithm to find optimally effective drug in a drug complex

Background The H1N1 pandemic in 2009 and the H5N1 pandemic in 2005 demonstrated that the drugs approved to treat influenza A viruses have low efficacy. This provided a stimulus for new studies of influenza A viruses in the context of the methods used in drug design developed over the past 100 years. Finding new universal drugs is the ultimate goal but its long time horizon is incompatible with emergency situations created by reoccurring influenza outbreaks. Therefore, we propose a computer-aided method for finding efficacious drugs and drug complexes based on the use of the DrugBank database. Methods (1) We start by assembling a panel of target proteins. (2) We then assemble a panel of drugs. (3) This is followed by a selection of benchmark binding pockets based on the panel of target proteins and the panel of drugs. (4) We generate a set of computational features, which measure the efficacy of a drug. (5) We propose a universal program to search for drugs and drug complexes. (6) A case study we report here illustrates how to use this universal program for finding an optimal drug and a drug complex for a given target. (7) Validation of the Azirchromycin and Aspirin complex is provided mathematically. (8) Finally, we propose a simple strategy to validate our computational prediction that the Azirchromycin and Aspirin complex should prove clinically effective. Result A set of computable features are mined and then based on these features, a universal program for finding the potential drug &drug complexes is proposed. Using this universal program, the Azirchromycin and Aspirin complex is selected and its efficacy is predicted mathematically. For clinical validation of this finding, future work is still required

Characterization and identification of the integrin family in silkworm, Bombyx mori

YesAs an important economic insect, Bombyx mori is also a useful model organism for lepidopteran insect. Integrins are evolutionarily conserved fromsponges to humans, and play vital roles inmany physiological and pathological processes. To explore their diverse functions of integrins in insect, eleven integrins including sixα and five β subunitswere cloned and characterized fromsilkworm. Our results showed that integrins fromsilkwormown more family members compared to other invertebrates. Among those α subunits, integrins α1, α2, and the other four subunits belong to PS1, PS2, and PS3 groups, respectively. The β subunits mainly gather in the insect βν group except the β1 subunit which belongs to the insect β group. Expression profiles demonstrated that the integrins exhibited distinct patterns, but were mainly expressed in hemocytes. α1 and β2 subunits are the predominant ones either in the embryogenesis or larva stages. Interestingly, integrins were significantly up-regulated after stimulated by 20-hydroxyecdysone (20-E) in vivo. These results indicate that integrins performdiverse functions in hemocytes of silkworm. Overall, our results provide a newinsight into the functional and evolutionary features of integrins.National Basic Research Programof China (No. 2012cb114603), the Research Fund for the Doctoral Program of Higher Education of China (20130182110003), the Natural Science Foundation of Chongqing (cstc2013jcyjys0007), and the Fundamental Research Funds for the Central Universities (SWU111014)

The Vertical Distribution of Ice-Nucleating Particles over the North China Plain: A Case of Cold Front Passage

Ice-nucleating particles (INPs) are crucial for cloud freezing processes in the atmosphere. Given the limited knowledge about the vertical distribution of INPs and its relation to aerosols in China, we present two aircraft observations of INPs over the North China Plain on 23 October 2019 and 25 October 2019, before and after a cold front passage. We used a well-established method to identify the INPs on a silicon wafer and then performed single-particle chemical composition analysis using an environmental scanning electron microscope-energy dispersive spectrometer (ESEM-EDS). The INP concentrations range from 0.1 to 9.2 L$^{−1}$ within activation temperatures from −20 to −29 °C. INPs are mostly concentrated within the boundary layer, and their concentration shows a decreasing trend with height (0.5~6 km) before the cold front passage. However, the highest INP concentration always appears at higher altitudes (4~5 km) after the cold front passage. The cold front passage also significantly weakens the correlations between the concentrations of INPs and aerosol particles at different sizes. The activated fraction (AF) of total aerosols increases from 10$^{−6}$ to 10$^{−4}$ with height from near ground to 6 km, reflecting a better nucleating capacity of the aerosols at higher altitudes. There is no obvious variation in AF after the cold front passage. Chemical analysis reveals that the INPs containing mineral dust components comprise the majority of total INPs during both flights. The proportion of pure mineral dust declines from 52.2% to 43.5% after the cold front passage while the proportion of mixed mineral dust increases from 23.9% to 45.7%, suggesting that an increased probability of aging or coating of INPs is introduced by the cold front during their long-distance transport. In addition, 88% of INPs have a diameter larger than 1 μm. This indicates that larger aerosols (>1 μm) are the major contributors to INPs at high altitudes despite their relatively low abundance. Our results demonstrate a significant impact of transport events on the sources and vertical distribution of INPs in the atmosphere