Background
Asthma is associated with inflammatory dysregulation, but the underlying metabolic signatures are unclear. This study aimed to classify asthma inflammatory phenotypes based on cellular and ...metabolic features.
Methods
To determine cellular and metabolic profiles, we assessed inflammatory cell markers using flow cytometry, sphingolipid (SL) metabolites using LC‐MS/MS, and serum cytokines using ELISA. Targeted gene polymorphisms were determined to identify genetic predispositions related to the asthma inflammatory phenotype.
Results
In total, 137 patients with asthma and 20 healthy controls (HCs) were enrolled. Distinct cellular and metabolic profiles were found between them; patients with asthma showed increased expressions of inflammatory cell markers and higher levels of SL metabolites compared to HCs (P < .05 for all). Cellular markers (CD66+ neutrophils, platelet‐adherent eosinophils) and SL metabolic markers (C16:0 and C24:0 ceramides) for uncontrolled asthma were also identified; higher levels were observed in uncontrolled asthma compared to controlled asthma (P < .05 for all). Asthmatics patients with higher levels of CD66+ neutrophils had lower FEV1(%), higher ACQ (but lower AQLO) scores, and higher sphingosine and C16:0 ceramide levels compared to those with low levels of CD66+ neutrophils. Asthmatics patients with higher levels of platelet‐adherent eosinophils had higher S1P levels compared to those with lower levels of platelet‐adherent eosinophils. Patients carrying TT genotype of ORMDL3 had more CD66+ neutrophils; those with AG/ GG genotypes of SGMS1 exhibited higher platelet‐adherent eosinophils.
Conclusion
Patients with uncontrolled asthma possess distinct inflammatory phenotypes including increased CD66+ neutrophils and platelet‐adherent eosinophils, with an imbalanced ceramide/S1P rheostat, potentially involving ORMDL3 and SGMS1 gene polymorphisms. Ceramide/S1P synthesis could be targeted to control airway inflammation.
Asthmatics patients with high level of CD66+ neutrophils have high C16:0 ceramide levels and enhanced ceramide‐mediated pro‐apoptotic signals (BID and JNK). Asthmatics patients with high level of platelet‐adherent eosinophils (CD61+ platelets/Siglec8+ eosinophils) have high S1P levels. The level of CD66+ neutrophils correlates with ROS production from neutrophils. The level of platelet‐adherent eosinophils is associated with asthma control symptom scores.
The FDA-approved small-molecule drug dasatinib is currently used as a treatment for chronic myeloid leukemia (CML). However, the effects of dasatinib on microglial and/or astrocytic neuroinflammatory ...responses and its mechanism of action have not been studied in detail.
BV2 microglial cells, primary astrocytes, or primary microglial cells were treated with dasatinib (100 or 250 nM) or vehicle (1% DMSO) for 30 min or 2 h followed by lipopolysaccharide (LPS; 200 ng/ml or 1 μg/ml) or PBS for 5.5 h. RT-PCR, real-time PCR; immunocytochemistry; subcellular fractionation; and immunohistochemistry were subsequently conducted to determine the effects of dasatinib on LPS-induced neuroinflammation. In addition, wild-type mice were injected with dasatinib (20 mg/kg, intraperitoneally (i.p.) daily for 4 days or 20 mg/kg, orally administered (p.o.) daily for 4 days or 2 weeks) or vehicle (4% DMSO + 30% polyethylene glycol (PEG) + 5% Tween 80), followed by injection with LPS (10 mg/kg, i.p.) or PBS. Then, immunohistochemistry was performed, and plasma IL-6, IL-1β, and TNF-α levels were analyzed by ELISA.
Dasatinib regulates LPS-induced proinflammatory cytokine and anti-inflammatory cytokine levels in BV2 microglial cells, primary microglial cells, and primary astrocytes. In BV2 microglial cells, dasatinib regulates LPS-induced proinflammatory cytokine levels by regulating TLR4/AKT and/or TLR4/ERK signaling. In addition, intraperitoneal injection and oral administration of dasatinib suppress LPS-induced microglial/astrocyte activation, proinflammatory cytokine levels (including brain and plasma levels), and neutrophil rolling in the brains of wild-type mice.
Our results suggest that dasatinib modulates LPS-induced microglial and astrocytic activation, proinflammatory cytokine levels, and neutrophil rolling in the brain.
Self-powered sensors have attracted significant interest for individual wearable device operation. Here, transparent and wearable single-electrode triboelectric nanogenerators (SETENGs) with high ...power generation are created using electrospun Ag nanowires (AgNWs)/poly(vinylidenefluoride-cotrifluoroethylene) P(VDF-TrFE) composite nanofibers (NFs). The SETENGs generate an output power density of up to 217 W/m2 with repetitive contact and separation from the surface of a latex glove. In electrospun P(VDF-TrFE) NFs, the crystalline β-phase is highly oriented by oxygen-containing functional groups on the surface of AgNWs, endowing the F-rich surface with high electron negativity and enabling efficient triboelectrification. Additionally, 80% transmittance at a light wavelength of 550 nm, mechanical stability, and durability after 10 000 cycles at 10% strain are confirmed by filling the NF pores with plasma desorption mass spectrometry. Our SETENG acts as an effective energy harvester by powering 45 light-emitting diodes and as an excellent real-time, self-powered touch panel.
Background
Obesity associated with various complications has increased worldwide. Body weight gain alters lipid metabolites (especially sphingolipids) contributing to obesity‐induced inflammation. ...However, the significance of the metabolites in the development of obese asthma is not yet clear.
Methods
The serum levels of sphingolipids were measured using liquid chromatography‐tandem mass spectrometry in obese controls (n = 7) and patients with asthma: the obese group (BMI > 25 kg/m2, n = 13) vs the nonobese (n = 28) group. To examine the relationship between metabolic changes in sphingolipids and macrophage polarization, public microarray data were analyzed. In addition, the alteration in sphingolipid metabolism was investigated in wild‐type BALB/c mice fed a high‐fat diet.
Results
The obese asthma had higher levels of serum C18:0 and C20:0 ceramides than the nonobese asthma group (P = .028 and P = .040, respectively). The value of the serum C18:0 ceramide (184.3 ng/mL) for discriminating the obese asthma from the nonobese asthma group showed 53.9% sensitivity and 85.7% specificity (AUC = 0.721, P = .024). The microarray data showed significantly increased ceramide synthesis and metabolic shift to ceramide accumulation during M1 macrophage polarization in humans. Increased airway hyperresponsiveness, M1 macrophage polarization, and C18:0 ceramide levels were noted in obese mice, but not in nonobese mice. Increased expression of ceramide synthase (CerS) 1 and CerS6 (not CerS2) was noted in lung tissues of obese mice.
Conclusion
Alteration in sphingolipid metabolism favoring ceramide accumulation (especially long‐chain ceramides) may contribute to developing obese asthma.
This study evaluates the association between sphingolipid metabolism and development of obese‐related asthma phenotype. Serum C18:0 ceramide is increased in patients with obese asthma. Gene expression involved in macrophage polarization may be associated with ceramide accumulation. Abbreviations: CERK, ceramide kinase; CERS6, ceramide synthase 6; PPAP2A, phosphatidic acid phosphatase type 2A; PPAP2B, phosphatidic acid phosphatase type 2B; SGMS2, sphingomyelin synthase 2; UGCG, UDP‐glucose ceramide glucosyltransferase.
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•Pluripotent stem cell (PSC)-derived expandable human hepatocyte-like liver organoids were generated.•PSC-derived human hepatic organoids are capable of long-term expansion with ...competent liver functionality.•PSC-derived human hepatic organoids provide a robust hepatic model for toxicity prediction and drug screening.
The development of hepatic models capable of long-term expansion with competent liver functionality is technically challenging in a personalized setting. Stem cell-based organoid technologies can provide an alternative source of patient-derived primary hepatocytes. However, self-renewing and functionally competent human pluripotent stem cell (PSC)-derived hepatic organoids have not been developed.
We developed a novel method to efficiently and reproducibly generate functionally mature human hepatic organoids derived from PSCs, including human embryonic stem cells and induced PSCs. The maturity of the organoids was validated by a detailed transcriptome analysis and functional performance assays. The organoids were applied to screening platforms for the prediction of toxicity and the evaluation of drugs that target hepatic steatosis through real-time monitoring of cellular bioenergetics and high-content analyses.
Our organoids were morphologically indistinguishable from adult liver tissue-derived epithelial organoids and exhibited self-renewal. With further maturation, their molecular features approximated those of liver tissue, although these features were lacking in 2D differentiated hepatocytes. Our organoids preserved mature liver properties, including serum protein production, drug metabolism and detoxifying functions, active mitochondrial bioenergetics, and regenerative and inflammatory responses. The organoids exhibited significant toxic responses to clinically relevant concentrations of drugs that had been withdrawn from the market due to hepatotoxicity and recapitulated human disease phenotypes such as hepatic steatosis.
Our organoids exhibit self-renewal (expandable and further able to differentiate) while maintaining their mature hepatic characteristics over long-term culture. These organoids may provide a versatile and valuable platform for physiologically and pathologically relevant hepatic models in the context of personalized medicine.
A functionally mature, human cell-based liver model exhibiting human responses in toxicity prediction and drug evaluation is urgently needed for pre-clinical drug development. Here, we develop a novel human pluripotent stem cell-derived hepatocyte-like liver organoid that is critically advanced in terms of its generation method, functional performance, and application technologies. Our organoids can contribute to the better understanding of liver development and regeneration, and provide insights for metabolic studies and disease modeling, as well as toxicity assessments and drug screening for personalized medicine.
G-protein-coupled receptor 40 (GPR40) is considered as an attractive drug target for treating type 2 diabetes, owing to its role in the free fatty acid-mediated increase in glucose-stimulated insulin ...secretion (GSIS) from pancreatic β-cells. To identify a new chemotype of GPR40 agonist, a series of 2-aryl-substituted indole-5-propanoic acid derivatives were designed and synthesized. We identified two GPR40 agonist lead compounds4k (3-2-(4-fluoro-2-methylphenyl)-1H-indol-5-ylpropanoic acid) and 4o (3-2-(2,5-dimethylphenyl)-1H-indol-5-ylpropanoic acid), having GSIS and glucagon-like peptide 1 secretory effects. Unlike previously reported GPR40 partial agonists that only activate the Gq pathway, 4k and 4o activated both the Gq and Gs signaling pathways and were characterized as GPR40 full agonists. In in vivo efficacy studies, 4o significantly improved glycemic control in both C57BL/6J and db/db mice and increased plasma-active GLP-1 in C57BL/6J mice. Thus, 4o represents a promising lead for further development as a novel GPR40 full agonist against type 2 diabetes.
In this paper, in order to develop the composition ceramics for piezoelectric actuator, (1-x)(Na
0.52
K
0.443
Li
0.037
)(Nb
0.883
Sb
0.08
Ta
0.037
)O
3
- x(Bi
0.5
Na
0.5
)
0.9
(Sr)
0.1
ZrO
3
ceramics ...were synthesized by the conventional solid state method and their piezoelectric and dielectric properties were systematically investigated. All specimens showed a typical pure perovskite structure and no secondary phase was found according to the (Bi
0.5
Na
0.5
)
0.9
(Sr)
0.1
ZrO
3
substitution. When x = 0.03, the excellent physical properties of d
33
= 256pC/N ε
r
= 1753, k
p
= 0.411, Qm = 58 and high Tc of 285 °C were obtained for piezoelectric actuator.
Down‐regulated surfactant protein B in obese asthmatics Cao, Thi Bich Tra; Moon, Ji‐Young; Yoo, Hyun‐Ju ...
Clinical and experimental allergy,
November 2022, 2022-11-00, 20221101, Letnik:
52, Številka:
11
Journal Article
Recenzirano
Background
Obesity is a common comorbid condition in adult asthmatics and known as a feature of asthma severity. However, the molecular mechanism under obesity‐induced inflammation has not yet been ...fully understood.
Objective
Considering the essential role of hydrophobic surfactant protein B (SP‐B) in lung function, SP‐B was targeted to examine its involvement in the development of obesity‐induced airway inflammation in asthmatics.
Methods
The aim was to examine an alteration in circulating SP‐B according to obesity in adult asthmatics, 129 asthmatics were enrolled and classified into 3 groups (obese, overweight and normal‐weight groups) according to body mass index (BMI). Circulating SP‐B levels were determined by enzyme‐linked immunosorbent assay. Four single nucleotide polymorphisms of SFTPB gene were genotyped. Serum ceramide levels were measured by liquid chromatography‐tandem mass spectrometry.
Results
Significantly lower serum SP‐B levels were noted in the obese group than in the overweight or normal‐weight group (p = .002). The serum SP‐B level was significantly correlated with serum levels of C18:0 ceramide and transforming growth factor beta 1 as well as BMI (r = −0.200; r = −0.215; r = −0.332, p < .050 for all). An inverse correlation was noted between serum SP‐B and fractional exhaled nitric oxide levels in female asthmatics (r = −0.287, p = .009). Genetic predisposition of the SFTPB gene at 9306 A>G to the obese and overweight groups was noted.
Conclusion
Obesity altered ceramide metabolism leading to pulmonary surfactant dysfunction and impaired resolution of airway inflammation, finally contributing to the phenotypes of obese asthmatics.
Considering the essential role of hydrophobic SP‐B in stabilizing the lipid layer on the surface of alveoli, dysregulated SP‐B was examined in obese subjects with asthma. The study found that down‐regulation of SP‐B had a significant correlation with increased levels of C18:0 ceramide/TGF‐β1/FeNO in obese asthmatics. The GG genotype frequency of SFTPB 9306 A>G polymorphism may be a genetic factor developing the phenotypes of obese asthma. Abbreviation: SP‐B, surfactant protein B; BMI, body mass index; FeNO, fractional exhaled nitric oxide; TGF‐β1, transforming growth factor beta 1.
The high circulating levels of histones found in various thrombotic diseases may compromise the anticoagulant barrier of endothelial cells. We determined how histones affect endothelial procoagulant ...tissue factor (TF) and anticoagulant thrombomodulin (TM). Surface antigens, soluble forms, and mRNA levels of TF and TM were measured by flow cytometry, ELISA, and real-time RT-PCR, respectively. TF and TM activity were measured using procoagulant activity, thrombin generation, or chromogenic assays. Involvement of the toll-like receptor (TLR) was assessed using the neutralizing antibodies. Histones dose-dependently induced surface antigens, activity and mRNA levels of endothelial TF. Histone-treated endothelial cells significantly shortened the lag time and enhanced the endogenous thrombin potential of normal plasma, which was normalized by a TF neutralizing antibody. Histones induced phosphatidylserine and protein-disulfide isomerase expression in endothelial cells. Histones also reduced the surface antigen, activity, and mRNA levels of endothelial TM. Polysialic acid and heparin reversed the histone-induced TF up-regulation and TM down-regulation. Activated protein C did not affect the TF up-regulation, but interrupted TM down-regulation. TLR2, and TLR4 inhibitors partially blocked the TF up-regulation. Histones induced the endothelial procoagulant phenotype through TF up-regulation and TM down-regulation. The effects of histones were partly mediated by TLR2, TLR4. Strategies to inhibit the harmful effects of histones in endothelial cells may be required in order to prevent a thrombotic environment.
Martensitic steels, engineered to meet automotive weight reduction and crashworthiness requirements, are highly susceptible to hydrogen embrittlement (HE). Despite extensive research to improve HE ...resistance, these steels remain vulnerable to HE after undergoing resistance spot welding (RSW). HE in spot-welded steels has been largely unexplored, and its underlying mechanism remains unclear. This study investigates the HE mechanism in RSWed steel sheets by examining microstructural alterations and hydrogen trapping sites before and after RSW. Additionally, the study analyzes variations in tensile-shear test (TST) properties relative to increasing hydrogen content. In spot-welded steels, the tensile-shear strength and fracture displacement decline as diffusible hydrogen levels increase. Specifically, fracture displacement sharply declines within the first 3 h of hydrogen charging, but this loss rate significantly decreases thereafter. This slope shift is attributed to a transition in the crack propagation path. Initially, cracks propagate along the fusion zone line, featuring transgranular failure. However, beyond a critical hydrogen content, crack propagation shifts to the prior austenite grain boundary (PAGB) in the upper critical heat-affected zone (UCHAZ), exhibiting intergranular failure. Consequently, when hydrogen content surpasses this critical threshold, the PAGB in the UCHAZ is weakened due to hydrogen-enhanced decohesion (HEDE) and hydrogen-enhanced localized plasticity (HELP). Cracks then propagate along the PAGB in the direction of maximum stress during the tensile-shear test (TST), leading to a change in the crack propagation path.
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•The resistance to hydrogen embrittlement for spot-welded advanced high strength steel is evaluated by tensile-shear test.•The difference in the behaviors of hydrogen embrittlement between steel matrix and welded part is compared.•The phenomenon of crack propagation path transition is observed over certain hydrogen content.•The mechanism of crack propagation path transition is discussed.
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