Chd4 choreographs self-antigen expression for central immune tolerance.

Autoreactive T cells are eliminated in the thymus to prevent autoimmunity by promiscuous expression of tissue-restricted self-antigens in medullary thymic epithelial cells. This expression is dependent on the transcription factor Fezf2, as well as the transcriptional regulator Aire, but the entire picture of the transcriptional program has been obscure. Here, we found that the chromatin remodeler Chd4, also called Mi-2ß, plays a key role in the self-antigen expression in medullary thymic epithelial cells. To maximize the diversity of self-antigen expression, Fezf2 and Aire utilized completely distinct transcriptional mechanisms, both of which were under the control of Chd4. Chd4 organized the promoter regions of Fezf2-dependent genes, while contributing to the Aire-mediated induction of self-antigens via super-enhancers. Mice deficient in Chd4 specifically in thymic epithelial cells exhibited autoimmune phenotypes, including T cell infiltration. Thus, Chd4 plays a critical role in integrating Fezf2- and Aire-mediated gene induction to establish central immune tolerance.

Characterization of an HNA aptamer suggests a non-canonical G-quadruplex motif.

Nucleic acids not only form the basis of heredity, but are increasingly a source of novel nano-structures, -devices and drugs. This has spurred the development of chemically modified alternatives (xeno nucleic acids (XNAs)) comprising chemical configurations not found in nature to extend their chemical and functional scope. XNAs can be evolved into ligands (XNA aptamers) that bind their targets with high affinity and specificity. However, detailed investigations into structural and functional aspects of XNA aptamers have been limited. Here we describe a detailed structure-function analysis of LYS-S8-19, a 1',5'-anhydrohexitol nucleic acid (HNA) aptamer to hen egg-white lysozyme (HEL). Mapping of the aptamer interaction interface with its cognate HEL target antigen revealed interaction epitopes, affinities, kinetics and hot-spots of binding energy similar to protein ligands such as anti-HEL-nanobodies. Truncation analysis and molecular dynamics (MD) simulations suggest that the HNA aptamer core motif folds into a novel and not previously observed HNA tertiary structure, comprising non-canonical hT-hA-hT/hT-hT-hT triplet and hG4-quadruplex structures, consistent with its recognition by two different G4-specific antibodies.

Leveraging Halogen Interactions for a Supramolecular Nanotube.

We demonstrate the formation of supramolecular nanotubes from molecular triangles in a single crystal by balancing the hydrogen bonds and halogen interactions between individual macrocycles. Thereby, we template the supramolecular nanotube growth by intermolecular interactions encoded directly in the macrocycles instead of those provided by the crystallization solvent. Ultimately, we show that replacing bromines for iodines in the macrocycle is necessary to achieve this supramolecular organization by enhancing the strength of the halogen interactions and concomitant reduction of the detrimental hydrogen bonds. We investigated the nature and the interplay of the individual intermolecular interactions by analysis of the experimental single crystal data and quantum chemical calculations. This work enriches the available toolbox of supramolecular interactions and will aid and abet the development of rationally-designed materials with a long-range 1D tubular organization.

Evaluation of the Antioxidant and Anti-Inflammatory Activities and Acute Toxicity of Caco Seed (Chrysobalanus icaco L.) in Murine Models.

Chysobalanus icaco L. (C. icaco) is a plant that is native to tropical America and Africa. It is also found in the southeast region of Mexico, where it is used as food and to treat certain diseases. This study aimed to carry out a phytochemical analysis of an aqueous extract of C. icaco seed (AECS), including its total phenol content (TPC), total flavonoid content (TFC), and condensed tannins (CT). It also aimed to examine the antioxidant and metal-ion-reducing potential of the AECS in vitro, as well as its toxicity and anti-inflammatory effect in mice. Antioxidant and metal-ion-reducing potential was examined by inhibiting DPPH, ABTS, and FRAP. The acute toxicity test involved a single administration of different doses of the AECS (0.5, 1, and 2 g/kg body weight). Finally, a single administration at doses of 150, 300, and 600 mg/kg of the AECS was used in the carrageenan-induced model of subplantar acute edema. The results showed that the AECS contained 124.14 ± 0.32 mg GAE, 1.65 ± 0.02 mg EQ, and 0.910 ± 0.01 mg of catechin equivalents/g dried extract (mg EC/g de extract) for TPC, TFC and CT, respectively. In the antioxidant potential assays, the values of the median inhibition concentration (IC50) of the AECS were determined with DPPH (0.050 mg/mL), ABTS (0.074 mg/mL), and FRAP (0.49 mg/mL). Acute toxicity testing of the AECS revealed no lethality, with a median lethal dose (LD50) value of >2 g/kg by the intragastric route. Finally, for inhibition of acute edema, the AECS decreased inflammation by 55%, similar to indomethacin (59%, p > 0.05). These results demonstrated that C. icaco seed could be considered a source of bioactive molecules for therapeutic purposes due to its antioxidant potential and anti-inflammatory activity derived from TPC, with no lethal effect from a single intragastric administration in mice.

Health-Related Composition and Bioactivity of an Agave Sap/Prickly Pear Juice Beverage.

In this study, a beverage made from a combination of Agave sap (AS) and prickly pear juice (PPJ) was analyzed for its nutrients and bioactive and potentially health-promoting compounds. The beverage was evaluated for its ability to act as an antioxidant, regulate glycemic properties, and undergo gut bacterial fermentation in vitro. The major mono- and oligosaccharides present in the beverage were galacturonic acid (217.74 ± 13.46 mg/100 mL), rhamnose (227.00 ± 1.58 mg/100 mL), and fructose (158.16 ± 8.86 mg/mL). The main phenolic compounds identified were protocatechuic acid (440.31 ± 3.06 mg/100 mL) and catechin (359.72 ± 7.56 mg/100 mL). It was observed that the beverage had a low glycemic index (<40) and could inhibit digestive carbohydrases. The combination of ingredients also helped to reduce gas production during AS fermentation from 56.77 cm3 to 15.67 cm3. The major SCFAs produced during fermentation were butyrate, acetate, and propionate, with valerate being produced only during the late fermentation of the AS. This beverage is rich in bioactive compounds, such as polyphenols and dietary fiber, which will bring health benefits when consumed.

Willingness to Participate in and Preferences for Studies of mHealth HIV Prevention Interventions: Cross-Sectional Study Among Sexual and Gender Minority Groups in the Southern United States.

A number of mobile HIV prevention interventions have been developed to increase uptake of HIV prevention services such as HIV testing and pre-exposure prophylaxis (PrEP). Most of these interventions have been tested among urban populations. However, sexual and gender minority (SGM) groups in rural areas might also benefit from mobile HIV prevention interventions. These groups have heightened experiences of stigma and discrimination and have limited access to culturally competent healthcare. We conducted a survey of SGM participants in the southern United States to assess willingness to use the common features of mobile HIV prevention interventions and to participate in research studies of these interventions and to compare the results between rural and non-rural respondents. We found few differences in willingness to use common features of mobile HIV prevention interventions based on rurality and high levels of cellular connectivity across participants. Based on these results, rural residence is not a barrier to using mobile HIV prevention interventions.

Disseminated intravascular coagulation phenotype is regulated by the TRPM7 channel during sepsis.

BACKGROUND: Sepsis is an uncontrolled inflammatory response against a systemic infection that results in elevated mortality, mainly induced by bacterial products known as endotoxins, producing endotoxemia. Disseminated intravascular coagulation (DIC) is frequently observed in septic patients and is associated with organ failure and death. Sepsis activates endothelial cells (ECs), promoting a prothrombotic phenotype contributing to DIC. Ion channel-mediated calcium permeability participates in coagulation. The transient reception potential melastatin 7 (TRPM7) non-selective divalent cation channel that also contains an α-kinase domain, which is permeable to divalent cations including Ca2+, regulates endotoxin-stimulated calcium permeability in ECs and is associated with increased mortality in septic patients. However, whether endothelial TRPM7 mediates endotoxemia-induced coagulation is not known. Therefore, our aim was to examine if TRPM7 mediates coagulation during endotoxemia. RESULTS: The results showed that TRPM7 regulated endotoxin-induced platelet and neutrophil adhesion to ECs, dependent on the TRPM7 ion channel activity and by the α-kinase function. Endotoxic animals showed that TRPM7 mediated neutrophil rolling on blood vessels and intravascular coagulation. TRPM7 mediated the increased expression of the adhesion proteins, von Willebrand factor (vWF), intercellular adhesion molecule 1 (ICAM-1), and P-selectin, which were also mediated by the TRPM7 α-kinase function. Notably, endotoxin-induced expression of vWF, ICAM-1 and P-selectin were required for endotoxin-induced platelet and neutrophil adhesion to ECs. Endotoxemic rats showed increased endothelial TRPM7 expression associated with a procoagulant phenotype, liver and kidney dysfunction, increased death events and an increased relative risk of death. Interestingly, circulating ECs (CECs) from septic shock patients (SSPs) showed increased TRPM7 expression associated with increased DIC scores and decreased survival times. Additionally, SSPs with a high expression of TRPM7 in CECs showed increased mortality and relative risk of death. Notably, CECs from SSPs showed significant results from the AUROC analyses for predicting mortality in SSPs that were better than the Acute Physiology and Chronic Health Evaluation II (APACHE II) and the Sequential Organ Failure Assessment (SOFA) scores. CONCLUSIONS: Our study demonstrates that sepsis-induced DIC is mediated by TRPM7 in ECs. TRPM7 ion channel activity and α-kinase function are required by DIC-mediated sepsis-induced organ dysfunction and its expression are associated with increased mortality during sepsis. TRPM7 appears as a new prognostic biomarker to predict mortality associated to DIC in SSPs, and as a novel target for drug development against DIC during infectious inflammatory diseases.

Antioxidant and Renin Inhibitory Activities of Peptides from Food Proteins on Hypertension: A Review.

Hypertension is a condition induced by oxidative stress causing an alteration in the endothelium, which increases the risk of suffering from other degenerative diseases. This review compiles the findings on peptides from food proteins with antioxidant and antihypertensive activities. Antihypertensive peptides are mainly focused on renin inhibition. Peptides containing hydrophobic amino acids have antioxidant and renin inhibitory activities, as reported by studies on the biological activity of peptides from various food sources evaluated separately and simultaneously. Peptides from food sources can present multiple biological activities. Moreover, antioxidant peptides have the potential to be evaluated against renin, offering an alternative for hypertension therapy without causing adverse side effects.

Distance as a barrier to HIV testing among sexual and gender minority populations in the rural southern US: a cross-sectional study.

INTRODUCTION: Sexual and gender minority people who live in rural areas are less likely to have had a HIV test in the previous 12 months compared with those who live in non-rural areas. We assessed the independent contribution of distance and time required to travel to receive a HIV test on recent uptake of HIV testing. METHODS: We conducted a cross-sectional survey of sexual and gender minority populations in the southern US. We used Poisson regression with robust standard errors to estimate prevalence ratios to compare uptake of HIV testing in the previous 12 months among those who traveled more than 20 miles (~32 km) and more than 30 minutes to their most recent HIV test compared with those who traveled less distance and time to their most recent test, respectively. RESULTS: A total of 508 (n=155 rural, n=348 non-rural) participants completed the survey. Of these, 398 (78.5%) had received a HIV test in the previous 12 months. Those who traveled more than 20 miles (~32 km) to their most recent test were more likely to have not received a HIV test in the previous 12 months compared with those who traveled 20 miles (~32 km) or less (adjusted prevalence ratio 2.25; 95% confidence interval 1.22-4.17). There were no differences based on travel time to the most recent test. CONCLUSION: Distance, but not time, to travel to receive a HIV test is independently associated with reduced HIV testing. More geographically proximal options or access to home-based testing might reduce this barrier.

Potential anti-inflammatory effects of legumes: a review.

Legumes are a staple of diets all around the world. In some least developed countries, they are the primary source of protein; however, their beneficial properties go beyond their nutritional value. Recent research has shown that legumes have bioactive compounds like peptides, polyphenols and saponins, which exhibit antioxidant, antihypertensive, anti-inflammatory and other biological activities. Thus, these compounds could be an alternative treatment for inflammatory diseases, in particular, chronic inflammation such as arthritis, obesity and cancer. Nowadays, there is a growing interest in alternative therapies derived from natural products; accordingly, the present review has compiled the bioactive compounds found in legumes that have demonstrated an anti-inflammatory effect in non-clinical studies.

Proteomic Analysis Reveals Differential Expression Profiles in Idiopathic Pulmonary Fibrosis Cell Lines.

Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, irreversible lung disorder of unknown cause. This disease is characterized by profibrotic activation of resident pulmonary fibroblasts resulting in aberrant deposition of extracellular matrix (ECM) proteins. However, although much is known about the pathophysiology of IPF, the cellular and molecular processes that occur and allow aberrant fibroblast activation remain an unmet need. To explore the differentially expressed proteins (DEPs) associated with aberrant activation of these fibroblasts, we used the IPF lung fibroblast cell lines LL97A (IPF-1) and LL29 (IPF-2), compared to the normal lung fibroblast cell line CCD19Lu (NL-1). Protein samples were quantified and identified using a label-free quantitative proteomic analysis approach by liquid chromatography-tandem mass spectrometry (LC-MS/MS). DEPs were identified after pairwise comparison, including all experimental groups. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG), and Protein-Protein Interaction (PPI) network construction were used to interpret the proteomic data. Eighty proteins expressed exclusively in the IPF-1 and IPF-2 clusters were identified. In addition, 19 proteins were identified up-regulated in IPF-1 and 10 in IPF-2; 10 proteins were down-regulated in IPF-1 and 2 in IPF-2 when compared to the NL-1 proteome. Using the search tool for retrieval of interacting genes/proteins (STRING) software, a PPI network was constructed between the DEPs and the 80 proteins expressed exclusively in the IPF-2 and IPF-1 clusters, containing 115 nodes and 136 edges. The 10 hub proteins present in the IPP network were identified using the CytoHubba plugin of the Cytoscape software. GO and KEGG pathway analyses showed that the hub proteins were mainly related to cell adhesion, integrin binding, and hematopoietic cell lineage. Our results provide relevant information on DEPs present in IPF lung fibroblast cell lines when compared to the normal lung fibroblast cell line that could play a key role during IPF pathogenesis.

Effect of the Instant Controlled Pressure Drop Technology in Cardamom (Elettaria cardamomum) Essential Oil Extraction and Antioxidant Activity.

Green cardamom (Elettaria cardamomum) is an outspread spice native to Asia, which is well appreciated for its sensory characteristics, delicate aroma, and unique taste. Currently, the main cardamom extracts are essential oils (EOs), and regarding current market tendencies, this market is in high growth. For this reason, technologies such as the instant controlled pressure drop (DIC) have been applied to reach higher yields and better quality of EO. Then, this study explores the impact of DIC as a pretreatment before hydrodistillation (HD) on the EO yield and their antioxidant activity. Obtained results showed that the coupling of DIC-HD increased the yield of essential oil and also had a positive impact on their antioxidant capacity. The EO yield of DIC-HD (140 °C and 30 s) was 4.43% vs. 2.52% for control; the AOX of DIC-HD (165 °C and 30 s) was 86% inhibition vs. 57.02% for control, and the TEAC of DIC-HD (140 °C and 30 s) was 1.44 uMTE/g EO vs. 13.66 uMTE/g EO.

DNA G-Quadruplex and i-Motif Structure Formation Is Interdependent in Human Cells.

Guanine- and cytosine-rich nucleic acid sequences have the potential to form secondary structures such as G-quadruplexes and i-motifs, respectively. We show that stabilization of G-quadruplexes using small molecules destabilizes the i-motifs, and vice versa, indicating these gene regulatory controllers are interdependent in human cells. This has important implications as these structures are predominately considered as isolated structural targets for therapy, but their interdependency highlights the interplay of both structures as an important gene regulatory switch.

Effect of Instant Controlled Pressure-Drop on the Non-Nutritional Compounds of Seeds and Sprouts of Common Black Bean (Phaseolus vulgaris L.).

The common bean is an important caloric-protein food source. However, its nutritional value may be affected by the presence of non-nutritional compounds, which decrease the assimilation of some nutrients; however, at low concentrations, they show a beneficial effect. Germination and treatment by controlled pressure-drop (DIC, French acronym of Détente Instantanée Contrôlée) are methods that modify the concentration of these components. The objective of this work was to evaluate the change in the non-nutritional composition of bean seeds and sprouts by DIC treatment. The results show that with the germination, the concentration of phenolic and tannin compounds increased 99% and 73%, respectively, as well as the quantity of saponins (65.7%), while phytates and trypsin inhibitors decreased 26% and 42%, respectively. When applying the DIC treatment, the content of phytates (23-29%), saponins (44%) and oligosaccharides increased in bean sprouts and decreased phenolic compounds (4-14%), tannins (23% to 72%), and trypsin inhibitors (95.5%), according to the pressure and time conditions applied. This technology opens the way to new perspectives, especially to more effective use of legumes as a source of vegetable protein or bioactive compounds.

Protective Effect of Chickpea Protein Hydrolysates on Colon Carcinogenesis Associated With a Hypercaloric Diet.

OBJECTIVE: Colon cancer occupies the third place in incidence worldwide; eating habits, in particular, consumption of hypercaloric diets, are relevant in its etiopathogenesis. On the other hand, foods can also modulate carcinogenesis: for example, proteins, which when hydrolyzed release peptides with biological activities, and legumes, especially, chickpea, represent a good source of hydrolysates. The objective of this work was to verify the inhibitory effect of chickpea hydrolyzed protein on azoxymethane (AOM)-induced carcinogenesis in mice fed a hypercaloric diet. METHODS: We hydrolyzed chickpea protein by pepsin, pancreatin, and a combined pepsin-pancreatin system, to test its anticarcinogenic and hypercaloric activity in mice that had consumed a hypercaloric diet or a normal diet but were injected with azoxymethane (AOM). RESULTS: A concentrate (70% proteins) was obtained from chickpea seeds (18.5% proteins), and extensive hydrolysates were obtained at 15 minutes, in all tested enzyme systems. The greatest activity was evidenced in the hydrolysates obtained with pepsin-pancreatin at 90 minutes. Animals that consumed the hypercaloric diet had a higher concentration of cholesterol and a higher atherogenic index, which were significantly reduced with the administration of chickpea protein hydrolysates with a dose-response effect (10, 20, or 30 mg/kg), whereas no effect was observed in animals that consumed the normal diet. In animals given AOM, aberrant crypts were observed, at a higher rate in animals that consumed the hypercaloric diet; with the consumption of hydrolysates by the animals that consumed either diet, the number of aberrant crypts was reduced with the 3 doses tested, and the effect was better in those animals fed the hypercaloric diet. The best effect in all tests was with 30 mg/kg body weight. CONCLUSION: The consumption of chickpea protein hydrolysates might confer a protective effect against colon carcinogenesis.

Fatigue in U12 Soccer-7 Players During Repeated 1-Day Tournament Games-A Pilot Study.

Sanchez-Sanchez, J, Sanchez, M, Hernandez, D, Ramirez-Campillo, R, Martínez, C, and Nakamura, FY. Fatigue in U12 Soccer-7 players during repeated 1-day tournament games-a pilot study. J Strength Cond Res 33(11): 3092-3097, 2019-The aim of this study was to describe and compare the distances and displacement speeds of U12 Soccer-7 athletes during 4 tournament Soccer-7 games (TG) played in less than 24 hours (experimental condition) with those recorded during 2 league Soccer-7 games (LG) with 24 hours of rest before the match (control condition). Ten participants (age = 10.3 ± 0.5 years) were recruited for the study. Main data analyzed during games included distance completed relative to match duration (Drel), maximal velocity, and distance completed at different running speeds (including acceleration, deceleration, standing, walking, jogging, medium-intensity running, high-intensity running, and sprinting). For data collection during games, athletes wore a global positioning system unit. Different (p ≤ 0.05) mean playing time was recorded during TG and LG (15.1 and 31.8 minutes/match, respectively). Drel during the 4 TG was maintained between 85.7 ± 8.5 and 87.5 ± 8.5 m·min (p > 0.05) and during the 2 LG between 84.2 ± 10.9 and 87.5 ± 9.9 m·min (p > 0.05). Moreover, similar Drel was recorded during TG and LG (86.8 and 85.9 m·min, respectively). Compared with LG, during TG, maximal velocity was lower (23.0 and 21.3 km·h, respectively; p ≤ 0.05). In addition, compared with the last game of the tournament, in the preceding games, the distance covered at low speeds (3.1-8.0 km·h) was lower (37.7 and 32.4%, respectively; p ≤ 0.05) and at high speeds (≥18.1 km·h) tends to be higher (2.5 and 3.3%, respectively). Therefore, compared with the control condition, accumulated Soccer-7 games with less than 24 hours of interday rest negatively affect displacement speed distribution (but not overall relative distances) in U12 Soccer-7 athletes. These results may help to better plan training and competition schedules to youth players.

New Molecular Insights into the Inhibition of Dipeptidyl Peptidase-4 by Natural Cyclic Peptide Oxytocin.

Protease inhibition has led to treating many diseases and has been successful in producing many commercial drugs by pharmaceutical companies. Among many proteases, serine protease has been attractive in treating metabolic disorder diabetes mellitus (DM). Gliptins have been proven to inhibit dipeptidyl peptidase-4 (DPP4), a serine protease, and are an emerging therapeutic drug target to reduce blood glucose levels, but until now there is no natural cyclic peptide proven to inhibit serine protease DPP4. This study demonstrates the potential mechanism of natural cyclic peptide oxytocin (OXT) as a DPP4 inhibitor. To achieve this, initially, activity atlas and field-based models of DPP4 inhibitors were utilized to predict the possible features of positive and negative electrostatic, hydrophobic, and activity shapes of DPP4 inhibition. Oxytocin binding mode, flexibility, and interacting residues were studied using molecular docking simulations studies. 3D-RISM calculations studies revealed that the stability of water molecules at the binding site are favorable. Finally, an experimental study using fluorescence assay revealed OXT inhibits DPP4 in a concentration-dependent manner in a significant way (p < 0.05) and possess IC50 of 110.7 nM. These new findings significantly expand the pharmaceutical application of cyclic peptides, and in specific OXT, and implicate further optimization of OXT inhibition capacity to understand the effect of DPP4 inhibition. This work highlights the development of natural cyclic peptides as future therapeutic peptides to reduce glucose levels and treat diabetes mellitus.

A Distance Bounding Protocol for Location-Cloaked Applications.

Location-based services (LBSs) assume that users are willing to release trustworthy and useful details about their whereabouts. However, many location privacy concerns have arisen. For location privacy protection, several algorithms build a cloaking region to hide a user’s location. However, many applications may not operate adequately on cloaked locations. For example, a traditional distance bounding protocol (DBP)—which is run by two nodes called the prover and the verifier—may conclude an untight and useless distance between these two entities. An LBS (verifier) may use this distance as a metric of usefulness and trustworthiness of the location claimed by the user (prover). However, we show that if a tight distance is desired, traditional DBP can refine a user’s cloaked location and compromise its location privacy. To find a proper balance, we propose a location-privacy-aware DBP protocol. Our solution consists of adding some small delays before submitting any user’s response. We show that several issues arise when a certain delay is chosen, and we propose some solutions. The effectiveness of our techniques in balancing location refinement and utility is demonstrated through simulation.

Trace element and toxic metal intake from the consumption of canned mushrooms marketed in Spain.

Mushrooms are a valuable foodstuff because they are an important source of fiber and essential amino acids, but they have a great capacity to capture metals from substrate. Due to this reason, the content of 14 metals (B, Ba, Cr, Cu, Fe, Li, Mo, Ni, Sr, V, Zn, Al, Cd, Pb) of 100 samples of canned mushrooms grown in Spain were analyzed by inductively coupled plasma atomic emission spectroscopy. Fe (9.55 mg/kg) was the most notable trace element, followed by Zn (2.39 mg/kg). The most abundant toxic metal was Al (17.80 mg/kg). Significant statistical differences between the absorption of metals have been observed when comparing species. Lentinula edodes and Lactarius deliciosus are the species which had the highest metal contents. Taking the average consumption 300 g per day for adults into account, the exposure to trace toxic metals from the consumption of the studied mushrooms does not pose a health risk. However, contribution percentages of Pb (87.6%) and Al (59.2%) found in Lentinula edodes and Pholiota nameko samples, respectively, are near the maximum intake.