Deficiency of neutral cholesterol ester hydrolase 1 (NCEH1) impairs endothelial function in diet-induced diabetic mice.

BACKGROUND: Neutral cholesterol ester hydrolase 1 (NCEH1) plays a critical role in the regulation of cholesterol ester metabolism. Deficiency of NCHE1 accelerated atherosclerotic lesion formation in mice. Nonetheless, the role of NCEH1 in endothelial dysfunction associated with diabetes has not been explored. The present study sought to investigate whether NCEH1 improved endothelial function in diabetes, and the underlying mechanisms were explored. METHODS: The expression and activity of NCEH1 were determined in obese mice with high-fat diet (HFD) feeding, high glucose (HG)-induced mouse aortae or primary endothelial cells (ECs). Endothelium-dependent relaxation (EDR) in aortae response to acetylcholine (Ach) was measured. RESULTS: Results showed that the expression and activity of NCEH1 were lower in HFD-induced mouse aortae, HG-exposed mouse aortae ex vivo, and HG-incubated primary ECs. HG exposure reduced EDR in mouse aortae, which was exaggerated by endothelial-specific deficiency of NCEH1, whereas NCEH1 overexpression restored the impaired EDR. Similar results were observed in HFD mice. Mechanically, NCEH1 ameliorated the disrupted EDR by dissociating endothelial nitric oxide synthase (eNOS) from caveolin-1 (Cav-1), leading to eNOS activation and nitric oxide (NO) release. Moreover, interaction of NCEH1 with the E3 ubiquitin-protein ligase ZNRF1 led to the degradation of Cav-1 through the ubiquitination pathway. Silencing Cav-1 and upregulating ZNRF1 were sufficient to improve EDR of diabetic aortas, while overexpression of Cav-1 and downregulation of ZNRF1 abolished the effects of NCEH1 on endothelial function in diabetes. Thus, NCEH1 preserves endothelial function through increasing NO bioavailability secondary to the disruption of the Cav-1/eNOS complex in the endothelium of diabetic mice, depending on ZNRF1-induced ubiquitination of Cav-1. CONCLUSIONS: NCEH1 may be a promising candidate for the prevention and treatment of vascular complications of diabetes.

Disrupted cardiac fibroblast BCAA catabolism contributes to diabetic cardiomyopathy via a periostin/NAP1L2/SIRT3 axis.

BACKGROUND: Periostin is an extracellular matrix protein that plays a critical role in cell fate determination and tissue remodeling, but the underlying role and mechanism of periostin in diabetic cardiomyopathy (DCM) are far from clear. Thus, we aimed to clarify the mechanistic participation of periostin in DCM. METHODS: The expression of periostin was examined in DCM patients, diabetic mice and high glucose (HG)-exposed cardiac fibroblasts (CF). Gain- and loss-of-function experiments assessed the potential role of periostin in DCM pathogenesis. RNA sequencing was used to investigate the underlying mechanisms of periostin in DCM. RESULTS: A mouse cytokine antibody array showed that the protein expression of periostin was most significantly upregulated in diabetic mouse heart, and this increase was also observed in patients with DCM or HG-incubated CF. Periostin-deficient mice were protected from diabetes-induced cardiac dysfunction and myocardial damage, while overexpression of periostin held the opposite effects. Hyperglycemia stimulated the expression of periostin in a TGF-ß/Smad-dependent manner. RNA sequencing results showed that periostin upregulated the expression of nucleosome assembly protein 1-like 2 (NAP1L2) which recruited SIRT3 to deacetylate H3K27ac on the promoters of the branched-chain amino acid (BCAA) catabolism-related enzymes BCAT2 and PP2Cm, resulting in BCAA catabolism impairment. Additionally, CF-derived periostin induced hypertrophy, oxidative injury and inflammation in primary cardiomyocytes. Finally, we identified that glucosyringic acid (GA) specifically targeted and inhibited periostin to ameliorate DCM. CONCLUSION: Overall, manipulating periostin expression may function as a promising strategy in the treatment of DCM.

Tetraspanins predict the prognosis and characterize the tumor immune microenvironment of glioblastoma.

Glioblastoma (GBM) is the most aggressive and lethal primary brain tumor. Conventional treatments have not achieved breakthroughs in improving survival. Therefore, novel molecular targets and biomarkers need to be identified. As signal transduction docks on the cell membrane, tetraspanins (TSPANs) are associated with various tumors; however, research on their role in GBM remains extremely scarce. Gene expression and clinicopathological characteristic data were obtained from GEPIA, CGGA, HPA, cBioPortal, and GSCA databases to analyze the mRNA and protein expression levels, prognostic value, clinical relevance, mutation status, and targeted drug sensitivity of TSPANs in GBM. Gene set enrichment analysis (GSEA), Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were used for biological process enrichment. Data from TCGA and TCIA were used to construct the tumor immune microenvironment landscape of TSPANs. Different R software algorithms were used to analyze the immune score, immune cell infiltration, and immune checkpoint correlation. Univariate and multivariate analyses were performed for TSPAN4, which had the most significant predictive prognostic value, and a nomogram model was constructed to predict individual outcomes. The expression and function of TSPAN4 were verified in vitro. TSPAN3/4/6/11/12/18/23/24/25/26/27/28/29/30/31expressions were significantly upregulated in GBM, and TSPAN3/4/6/11/18/24/25/26/29/30 were strongly correlated with prognosis. The expression of multiple TSPANs significantly correlated with 1p/19q co-deletion status, IDH mutation status, recurrence, age, and tumor grade. GSEA and GO analyses revealed the potential contribution of TSPANs in cell adhesion and migration. Immune correlation analysis revealed that TSPANs are related to the formation of the GBM tumor microenvironment (TME) and may influence immunotherapy outcomes. TSPAN4 is an independent prognostic factor and TSPAN4 knockdown has been demonstrated to strongly inhibit glioma cell proliferation, invasion, and migration in vitro. We comprehensively elaborated the prognostic value and potential role of differentially expressed TSPANs in GBM, including molecules that scientists have previously overlooked. This study provides a novel and comprehensive perspective on the pathological mechanisms of GBM and the future direction of individualized tumor immunotherapy, which may be a critical link between GBM malignant progression and TME remodeling.

Rheological Role of Stiff Nanorings on Concurrently Strengthening and Toughening Polymer Nanocomposites.

Nanorings, which are increasingly uncovered in natural systems and synthesized in man-made materials, exhibit dynamics distinct from those known for linear chains. We show in this study that, when immersed in a polymer melt matrix, segments of a stiff nanoring (SNR) have more facilitated subdiffusion, i.e., with a larger scaling exponent in the mean squared displacement, than those belonging to one flexible counterpart, while the whole SNR is more suppressed by its surroundings. It is revealed that adding SNRs contributes to achieving the long-anticipated rheological objective of sol- and gel-like characteristics at high and low shearing frequencies, respectively. This study suggests the promising prospect of exploiting SNRs to concurrently strengthen and toughen target polymer nanocomposites.

SPP1/AnxA1/TIMP1 as Essential Genes Regulate the Inflammatory Response in the Acute Phase of Cerebral Ischemia-Reperfusion in Rats.

Background: Ischemic injury in stroke is followed by extensive neurovascular inflammation and changes in ischemic penumbra gene expression patterns. However, the key molecules involved in the inflammatory response during the acute phase of ischemic stroke remain unclear. Methods: Gene expression profiles of two rat ischemic stroke-related data sets, GSE61616 and GSE97537, were downloaded from the GEO database for Gene Set Enrichment Analysis (GSEA). Then, GEO2R was used to screen differentially expressed genes (DEGs). Furthermore, 170 differentially expressed intersection genes were screened and analyzed for Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Candidate genes and miRNAs were obtained by DAVID, Metascape, Cytoscape, STRING, and TargetScan. Finally, the rat middle cerebral artery occlusion-reperfusion (MCAO/R) model was constructed, and qRT-PCR was used to verify the predicted potential miRNA molecule and its target genes. Results: GO and KEGG analyses showed that 170 genes were highly associated with inflammatory cell activation and cytokine production. After cluster analysis, seven hub genes highly correlated with post-stroke neuroinflammation were obtained: Cxcl1, Kng1, Il6, AnxA1, TIMP1, SPP1, and Ccl6. The results of TargetScan further suggested that miR-340-5p may negatively regulate SPP1, AnxA1, and TIMP1 simultaneously. In the ischemic penumbra of rats 24 h after MCAO/R, the level of miR-340-5p significantly decreased compared with the control group, while the concentration of SPP1, AnxA1, and TIMP1 increased. Time-course studies demonstrated that the mRNA expression levels of SPP1, AnxA1, and TIMP1 fluctuated dramatically throughout the acute phase of cerebral ischemia-reperfusion (I/R). Conclusion: Our study suggests that differentially expressed genes SPP1, TIMP1, and ANXA1 may play a vital role in the inflammatory response during the acute phase of cerebral ischemia-reperfusion injury. These genes may be negatively regulated by miR-340-5p. Our results may provide new insights into the complex pathophysiological mechanisms of secondary inflammation after stroke.

Exploration on the Mechanism of Ubiquitin Proteasome System in Cerebral Stroke.

Stroke's secondary damage, such as inflammation, oxidative stress, and mitochondrial dysfunction, are thought to be crucial factors in the disease's progression. Despite the fact that there are numerous treatments for secondary damage following stroke, such as antiplatelet therapy, anticoagulant therapy, surgery, and so on, the results are disappointing and the side effects are numerous. It is critical to develop novel and effective strategies for improving patient prognosis. The ubiquitin proteasome system (UPS) is the hub for the processing and metabolism of a wide range of functional regulatory proteins in cells. It is critical for the maintenance of cell homeostasis. With the advancement of UPS research in recent years, it has been discovered that UPS is engaged in a variety of physiological and pathological processes in the human body. UPS is expected to play a role in the onset and progression of stroke via multiple targets and pathways. This paper explores the method by which UPS participates in the linked pathogenic process following stroke, in order to give a theoretical foundation for further research into UPS and stroke treatment.

[Research Progress in the Association between Amino Acid Metabolism of Oral Microorganisms and Host Cells and Oral Diseases].

Amino acids, the substrate of protein synthesis, are an important source of energy and nutrition, second only to glucose. Previous studies have found that both microorganisms and their host cells can metabolize amino acids, and the metabolites are widely involved in the regulation of various biological processes, including inflammation and immune response. Exploring the changes in amino acid metabolism during the pathogenesis and progression of diseases has become a new hot topic of research. We summarized in this review the research progress in the pathogenesis and progression of common oral diseases, including dental caries, periodontal diseases, Sjögren's syndrome, and even oral tumors, related to metabolism pathways of amino acids, especially tryptophan and arginine, and their metabolites, attempting to provide a theoretical basis for enhancing understanding of the pathogenic mechanism of the oral diseases, as well as guidance for clinical treatment.

Measurement of contaminant adsorption on soils using cycling modified column tests.

An innovative cycled column test with supporting batch equilibrium and kinetic analysis for adsorption or desorption were developed for evaluation of adsorption behavior of soils. Non-equilibrium adsorption was observed in the cycled column tests as the traditional testing methods. The isotherm of local equilibrium of the soil was conducted based on the testing results within a relatively short duration with simple analysis. The concentration curves of influent and effluent of the cycled column tests were simulated by dual-porosity (DP) model with a modified inlet boundary. Based on the modeling results, the isotherm of local equilibrium is close to that of the mobile phase adsorption capacity, whereas the immobile phase of the soil is nearly inactive in the retardation of the contaminants. The testing results from cycled column tests are hardly interfered by desorption or the sorption rate according to the modeling for corresponding scenarios. The cycled column test can be used as an alternative or supplementary method to the traditional column test for the determination of local equilibrium isotherm, with advantages of shorter testing duration and easier data analysis.

Photonic Nanojet-Mediated Optogenetics.

Optogenetics has become a widely used technique in neuroscience research, capable of controlling neuronal activity with high spatiotemporal precision and cell-type specificity. Expressing exogenous opsins in the selected cells can induce neuronal activation upon light irradiation, and the activation depends on the power of incident light. However, high optical power can also lead to off-target neuronal activation or even cell damage. Limiting the incident power, but enhancing power distribution to the targeted neurons, can improve optogenetic efficiency and reduce off-target effects. Here, the use of optical lenses made of polystyrene microspheres is demonstrated to achieve effective focusing of the incident light of relatively low power to neighboring neurons via photonic jets. The presence of microspheres significantly localizes and enhances the power density to the target neurons both in vitro and ex vivo, resulting in increased inward current and evoked action potentials. In vivo results show optogenetic stimulation with microspheres that can evoke significantly more motor behavior and neuronal activation at lowered power density. In all, a proof-of-concept of a strategy is demonstrated to increase the efficacy of optogenetic neuromodulation using pulses of reduced optical power.

Honeycomb-patterned porous films fabricated via self-organization of Tb complex-loaded amphiphilic copolymers.

Amphiphilic copolymers, poly(styrene)-block-Tb complex (PS-b-Tb complex), were synthesized by reversible addition fragmentation chain transfer (RAFT) polymerization. The honeycomb structured porous films were fabricated via dropping the PS-b-Tb complex copolymer solutions on glass substrates by the breath figures method (BFM). The structure and composition of the amphiphilic copolymer PS-b-Tb complex were confirmed by gel permeation chromatography (GPC), Fourier transform infrared spectroscopy (FT-IR) and 1H nuclear magnetic resonance spectroscopy (1H NMR). The surface morphology and elemental mapping of the highly ordered porous films were investigated by field emission scanning electron microscopy (FESEM), energy dispersive X-ray spectroscopy (EDX) and laser scanning confocal microscopy (LSCM). The results indicated that the solvent type and copolymer concentration can affect the surface morphology of the porous films. The average diameter of the pores in the porous films decreased with the polymer concentration and the molecular weight of the copolymers increased. The FESEM-EDX analysis further verified that the hydrophilic groups (Tb complex groups) were mainly distributed at the pore wall, instead of at the outer surface layer of the films, which was consistent with the LSCM results.

A degradation model for high kitchen waste content municipal solid waste.

Municipal solid waste (MSW) in developing countries has a high content of kitchen waste (KW), and therefore contains large quantities of water and non-hollocellulose degradable organics. The degradation of high KW content MSW cannot be well simulated by the existing degradation models, which are mostly established for low KW content MSW in developed countries. This paper presents a two-stage anaerobic degradation model for high KW content MSW with degradations of hollocellulose, sugars, proteins and lipids considered. The ranges of the proportions of chemical compounds in MSW components are summarized with the recommended values given. Waste components are grouped into rapidly or slowly degradable categories in terms of the degradation rates under optimal water conditions for degradation. In the proposed model, the unionized VFA inhibitions of hydrolysis/acidogenesis and methanogenesis are considered as well as the pH inhibition of methanogenesis. Both modest and serious VFA inhibitions can be modeled by the proposed model. Default values for the parameters in the proposed method can be used for predictions of degradations of both low and high KW content MSW. The proposed model was verified by simulating two laboratory experiments, in which low and high KW content MSW were used, respectively. The simulated results are in good agreement with the measured data of the experiments. The results show that under low VFA concentrations, the pH inhibition of methanogenesis is the main inhibition to be considered, while the inhibitions of both hydrolysis/acidogenesis and methanogenesis caused by unionized VFA are significant under high VFA concentrations. The model is also used to compare the degradation behaviors of low and high KW content MSW under a favorable environmental condition, and it shows that the gas potential of high KW content MSW releases more quickly.

Manipulation and detection of single nanoparticles and biomolecules by a photonic nanojet.

Optical methods to manipulate and detect nanoscale objects are highly desired in both nanomaterials and molecular biology fields. Optical tweezers have been used to manipulate objects that range in size from a few hundred nanometres to several micrometres. The emergence of near-field methods that overcome the diffraction limit has enabled the manipulation of objects below 100 nm. A highly free manipulation with signal-enhanced real-time detection, however, remains a challenge for single sub-100-nm nanoparticles or biomolecules. Here we show an approach that uses a photonic nanojet to perform the manipulation and detection of single sub-100-nm objects. With the photonic nanojet generated by a dielectric microlens bound to an optical fibre probe, three-dimensional manipulations were achieved for a single 85-nm fluorescent polystyrene nanoparticle as well as for a plasmid DNA molecule. Backscattering and fluorescent signals were detected with the enhancement factors up to ∼103 and ∼30, respectively. The demonstrated approach provides a potentially powerful tool for nanostructure assembly, biosensing and single-biomolecule studies.

Fabrication of honeycomb-patterned porous films from PS-b-PNIPAM amphiphilic diblock copolymers synthesized via RITP.

Poly(styrene)-b-poly(N-isopropylacrylamide) (PS-b-PNIPAM) amphiphilic diblock copolymers were synthesized via reverse iodine transfer polymerization (RITP) firstly. And then the honeycomb structured porous films were fabricated by drop the amphiphilic diblock copolymer solutions on glass substrates. The porous films were characterized using field emission scanning electron microscopy (FESEM) and atomic force microscopy (AFM), respectively. The results showed that the regular honeycomb patterned porous film was achieved successfully by employing the block copolymer with a significantly larger polystyrene block proportion into the solution and the pore diameter of the film decreases obviously with the polymer concentration increased. The effects of airflow speed and Ag particles on porous films are also discussed in this study.

Influences of operational practices on municipal solid waste landfill storage capacity.

The quantitative effects of three operational factors, that is initial compaction, decomposition condition and leachate level, on municipal solid waste (MSW) landfill settlement and storage capacity are investigated in this article via consideration of a hypothetical case. The implemented model for calculating landfill compression displacement is able to consider decreases in compressibility induced by biological decomposition and load dependence of decomposition compression for the MSW. According to the investigation, a significant increase in storage capacity can be achieved by intensive initial compaction, adjustment of decomposition condition and lowering of leachate levels. The quantitative investigation presented aims to encourage landfill operators to improve management to enhance storage capacity. Furthermore, improving initial compaction and creating a preferential decomposition condition can also significantly reduce operational and post-closure settlements, respectively, which helps protect leachate and gas management infrastructure and monitoring equipment in modern landfills.

[Impacts of initial moisture content of MSW waste on leachate generation and modified formula for predicting leachate generation].

The amount of leachate generation rate in MSW landfills is often underestimated during design phase in China. A water balance model of a valley landfill, whose size is 400 m long, 500 m wide and 50 m thick, is created to investigate the influences of initial moisture content of waste on source and production of leachate. The 50 m thick waste mass is assumed to be 5 layers. Each layer is 10 m thick with a filling period of 2 years. The leachate mainly comes from precipitation and from squeezed pore water of waste. It is found that higher initial moisture content of waste leads to higher amounts of squeezed leachate and total leachate generation rate, and also results in a high ratio of squeezed leachate to total leachate generation rate. For the cases that the initial moisture contents of waste are 27%, 40%, 50%, and 60%, the amounts of total leachate generation rate are 272, 583, 823 and 1 063 m3 x d(-1), respectively, and the amounts of squeezed leachate are--144, 168, 408, and 647 m3 x d(-1), respectively. It is also found that when the initial moisture content of waste is greater than 50%, the squeezed leachate becomes the primary source of total leachate generation rate. However, the formula for predicting leachate generation rate used in the national code could not consider the contribution of squeezed leachate, this may cause a significant underestimation of leachate generation rate for the case having a high initial moisture content of waste. Based on the water balance analyses, a modified formula for predicting leachate generation rate, which includes the contribution of squeezed leachate is proposed. It is verified by consideration of the operational practices of two large-scale landfills in southern China.

Fabrication of block copolymer brushes on hollow sphere surface via reverse iodine transfer polymerization.

The block copolymer brushes grafted from hollow sphere surface via reverse iodine transfer polymerization (RITP) were investigated in this work. A sufficient amount of azo initiator was introduced onto hollow sphere surface firstly. Then the monomer methyl methacrylate (MMA) was polymerized via surface-initiated reverse iodine transfer polymerization (RITP) using azo group modified hollow sphere as initiator. The microstructure of the samples was characterized by FT-IR, (1)H NMR, respectively. Results indicated that the poly(methyl methacrylate) (PMMA) with end functionality of alkyl iodine group had grafted from hollow sphere surface. TEM observations showed that the average diameter of hollow core was central at 1.3-1.4 µm and the average wall thickness increased from 103 nm to 138 nm and 172 nm after grafting polymerization of MMA and Tb complex, respectively. The closely linear plots of molecular weight (M(n)) versus conversion, linear kinetic plots for the free polymer formed in solution and the ability to extend the chains by sequential addition of monomer indicated that the RITP was a controlled process with a "living" characteristic.