Study on the Performance of Epoxy-Modified Asphalt and Steel Slag Ultra-Thin Friction Course.

Ultra-thin overlays (UTOL) are a standard highway pre-maintenance method used to improve the road surface performance of asphalt pavements and to repair minor rutting and cracking. However, the thin thickness makes it very sensitive to external changes, which increases its wear and shortens its life. So, this paper aims to prepare a durable and skid-resistance asphalt ultra-thin overlay using epoxy asphalt (EA) and steel slag. First, the physical properties of EA were characterized by penetration, softening point, flexibility, and kinematic viscosity tests. The dynamic shear rheometer (DSR) test characterizes EA's rheological properties. Differential Scanning Calorimetry (DSC), kinematic viscosity, and Fourier transform infrared spectroscopy (FTIR) characterized the EA's curing process. Finally, the pavement performance of an epoxy ultra-thin overlay (EUTOL) prepared with EA and steel slag was tested. The results show that the epoxy resin particles increase with the increase in epoxy resin dosage, and at 40%, its epoxy particles are uniformly distributed with the most significant area share. With the addition of epoxy resin, the needle penetration of EA decreases and then increases, the flexibility decreases at a slower rate, and the softening point rises significantly. Moreover, the growth of the elastic component in EA significantly improved the high-temperature viscoelastic properties. Considering its physical and rheological properties, the optimal doping amount of 40% was selected. By analyzing the curing behavior of EA (optimum dosage), the combination temperature of EA is 150 °C, which meets the needs of mixing and paving asphalt mixtures. After 12 h of maintenance at 120 °C, its reaction is sufficient. The skid-resistance durability, high-temperature, low-temperature, water stability, and fatigue resistance of UTOL can be effectively improved using steel slag coarse aggregate.

TRP Channels in Excitotoxicity.

Glutamate excitotoxicity is a central mechanism contributing to cellular dysfunction and death in various neurological disorders and diseases, such as stroke, traumatic brain injury, epilepsy, schizophrenia, addiction, mood disorders, Huntington's disease, Alzheimer's disease, Parkinson's disease, multiple sclerosis, pathologic pain, and even normal aging-related changes. This detrimental effect emerges from glutamate binding to glutamate receptors, including α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors, N-methyl-d-aspartate receptors, kainate receptors, and GluD receptors. Thus, excitotoxicity could be prevented by targeting glutamate receptors and their downstream signaling pathways. However, almost all the glutamate receptor antagonists failed to attenuate excitotoxicity in human patients, mainly due to the limited understanding of the underlying mechanisms regulating excitotoxicity. Transient receptor potential (TRP) channels serve as ancient cellular sensors capable of detecting and responding to both external and internal stimuli. The study of human TRP channels has flourished in recent decades since the initial discovery of mammalian TRP in 1995. These channels have been found to play pivotal roles in numerous pathologic conditions, including excitotoxicity. In this review, our focus centers on exploring the intricate interactions between TRP channels and glutamate receptors in excitotoxicity.

TRPM2 enhances ischemic excitotoxicity by associating with PKCγ.

N-methyl-D-aspartate receptor (NMDAR)-mediated glutamate excitotoxicity significantly contributes to ischemic neuronal death and post-recanalization infarction expansion. Despite tremendous efforts, targeting NMDARs has proven unsuccessful in clinical trials for mitigating brain injury. Here, we show the discovery of an interaction motif for transient receptor potential melastatin 2 (TRPM2) and protein kinase Cγ (PKCγ) association and demonstrate that TRPM2-PKCγ uncoupling is an effective therapeutic strategy for attenuating NMDAR-mediated excitotoxicity in ischemic stroke. We demonstrate that the TRPM2-PKCγ interaction allows TRPM2-mediated Ca2+ influx to promote PKCγ activation, which subsequently enhances TRPM2-induced potentiation of extrasynaptic NMDAR (esNMDAR) activity. By identifying the PKCγ binding motif on TRPM2 (M2PBM), which directly associates with the C2 domain of PKCγ, an interfering peptide (TAT-M2PBM) is developed to disrupt TRPM2-PKCγ interaction without compromising PKCγ function. M2PBM deletion or TRPM2-PKCγ dissociation abolishes both TRPM2-PKCγ and TRPM2-esNMDAR couplings, resulting in reduced excitotoxic neuronal death and attenuated ischemic brain injury.

Activation of endothelial TRPM2 exacerbates blood-brain barrier degradation in ischemic stroke.

AIMS: Damage of the blood-brain barrier (BBB) is a hallmark of brain injury during the early stages of ischemic stroke. The subsequent endothelial hyperpermeability drives the initial pathological changes and aggravates neuronal death. Transient receptor potential melastatin 2 (TRPM2) is a Ca2+-permeable nonselective cation channel activated by oxidative stress. However, whether TRPM2 is involved in BBB degradation during ischemic stroke remains unknown. We aimed to investigate the role of TRPM2 in BBB degradation during ischemic stroke and the underlying molecular mechanisms. METHODS AND RESULTS: Specific deletion of Trpm2 in endothelial cells using Cdh5 Cre produces a potent protective effect against brain injury in mice subjected to middle cerebral artery occlusion (MCAO), which is characterized by reduced infarction size, mitigated plasma extravasation, suppressed immune cell invasion, and inhibited oxidative stress. In vitro experiments using cultured cerebral endothelial cells (CECs) demonstrated that either Trpm2 deletion or inhibition of TRPM2 activation attenuates oxidative stress, Ca2+ overload, and endothelial hyperpermeability induced by oxygen-glucose deprivation (OGD) and CD36 ligand thrombospondin-1 (TSP1). In transfected HEK293T cells, OGD and TSP1 activate TRPM2 in a CD36-dependent manner. Noticeably, in cultured CECs, deleting Trpm2 or inhibiting TRPM2 activation also suppresses the activation of CD36 and cellular dysfunction induced by OGD or TSP1. CONCLUSIONS: In conclusion, our data reveal a novel molecular mechanism in which TRPM2 and CD36 promote the activation of each other, which exacerbates endothelial dysfunction during ischemic stroke. Our study suggests that TRPM2 in endothelial cells is a promising target for developing more effective and safer therapies for ischemic stroke.

TRP Channels in Stroke.

Ischemic stroke is a devastating disease that affects millions of patients worldwide. Unfortunately, there are no effective medications for mitigating brain injury after ischemic stroke. TRP channels are evolutionally ancient biosensors that detect external stimuli as well as tissue or cellular injury. To date, many members of the TRP superfamily have been reported to contribute to ischemic brain injury, including the TRPC subfamily (1, 3, 4, 5, 6, 7), TRPV subfamily (1, 2, 3, 4) and TRPM subfamily (2, 4, 7). These TRP channels share structural similarities but have distinct channel functions and properties. Their activation during ischemic stroke can be beneficial, detrimental, or even both. In this review, we focus on discussing the interesting features of stroke-related TRP channels and summarizing the underlying cellular and molecular mechanisms responsible for their involvement in ischemic brain injury.

Cardiac ultrastructure inspired matrix induces advanced metabolic and functional maturation of differentiated human cardiomyocytes.

The vast potential of human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) in preclinical models of cardiac pathologies, precision medicine, and drug screening remains to be fully realized because hiPSC-CMs are immature without adult-like characteristics. Here, we present a method to accelerate hiPSC-CM maturation on a substrate, cardiac mimetic matrix (CMM), mimicking adult human heart matrix ligand chemistry, rigidity, and submicron ultrastructure, which synergistically mature hiPSC-CMs rapidly within 30 days. hiPSC-CMs matured on CMM exhibit systemic transcriptomic maturation toward an adult heart state, are aligned with high strain energy, metabolically rely on oxidative phosphorylation and fatty acid oxidation, and display enhanced redox handling capability, efficient calcium handling, and electrophysiological features of ventricular myocytes. Endothelin-1-induced pathological hypertrophy is mitigated on CMM, highlighting the role of a native cardiac microenvironment in withstanding hypertrophy progression. CMM is a convenient model for accelerated development of ventricular myocytes manifesting highly specialized cardiac-specific functions.

Research on Design and Performance of Self-Compacting Cement Emulsified Bitumen Mixture (CEBM).

To meet the needs of the road industry for maintenance operations, a new cement emulsified bitumen mixture (CEBM) with early-strength, self-compacting, and room-temperature construction characteristics was designed. The strength formation mechanism of CEBM was revealed with a scanning electron microscope (SEM) and the surface free energy (SFE) theory. The mechanical properties and road performance of the CEBM were investigated extensively. The results show that before the demulsification of emulsified bitumen, the SFE of the bitumen-aggregate-water three-phase system was reduced due to the replacement of the bitumen-aggregate interface with water. The adhesion work between the emulsified bitumen and the aggregate is negative, which means the adhesion between the emulsified bitumen and the aggregate will not occur spontaneously due to the existence of water. The liquid emulsified bitumen improves the workability of the mixture and ensures that the mixture can be evenly mixed and self-compacted. After demulsification, the work of adhesion between the residual bitumen and the aggregate is positive, which means residual bitumen and aggregate can bond spontaneously. In addition, the hydration products of cement and aggregate form a skeleton, and the emulsified bitumen film wraps and bonds the cement and aggregate together, creating strength. The emulsified bitumen, cement content, and curing conditions have significant effects on the stability of CEBM. The recommended dosage of emulsified bitumen and cement is 8% and 8-10%, respectively. This material integrates the hardening effect of cement and the viscoelastic performance of bitumen and has good workability, mechanical properties, and road performance. Therefore, the CEBM is technically feasible for application to bitumen pavement.

Functional coupling of TRPM2 and extrasynaptic NMDARs exacerbates excitotoxicity in ischemic brain injury.

Excitotoxicity induced by NMDA receptor (NMDAR) activation is a major cause of neuronal death in ischemic stroke. However, past efforts of directly targeting NMDARs have unfortunately failed in clinical trials. Here, we reveal an unexpected mechanism underlying NMDAR-mediated neurotoxicity, which leads to the identification of a novel target and development of an effective therapeutic peptide for ischemic stroke. We show that NMDAR-induced excitotoxicity is enhanced by physical and functional coupling of NMDAR to an ion channel TRPM2 upon ischemic insults. TRPM2-NMDAR association promotes the surface expression of extrasynaptic NMDARs, leading to enhanced NMDAR activity and increased neuronal death. We identified a specific NMDAR-interacting motif on TRPM2 and designed a membrane-permeable peptide to uncouple the TRPM2-NMDAR interaction. This disrupting peptide protects neurons against ischemic injury in vitro and protects mice against ischemic stroke in vivo. These findings provide an unconventional strategy to mitigate excitotoxic neuronal death without directly targeting NMDARs.

TRPM2 deficiency in mice protects against atherosclerosis by inhibiting TRPM2-CD36 inflammatory axis in macrophages.

Atherosclerosis is the major cause of ischemic heart disease and stroke, the leading causes of mortality worldwide. The central pathological features of atherosclerosis include macrophage infiltration and foam cell formation. However, the detailed mechanisms regulating these two processes remain unclear. Here we show that oxidative stress-activated Ca2+-permeable transient receptor potential melastatin 2 (TRPM2) plays a critical role in atherogenesis. Both global and macrophage-specific Trpm2 deletion protect Apoe -/- mice against atherosclerosis. Trpm2 deficiency reduces oxidized low-density lipoprotein (oxLDL) uptake by macrophages, thereby minimizing macrophage infiltration, foam cell formation and inflammatory responses. Activation of the oxLDL receptor CD36 induces TRPM2 activity, and vice versa. In cultured macrophages, TRPM2 is activated by CD36 ligands oxLDL and thrombospondin-1 (TSP1), and deleting Trpm2 or inhibiting TRPM2 activity suppresses the activation of CD36 signaling cascade induced by oxLDL and TSP1. Our findings establish the TRPM2-CD36 axis as a molecular mechanism underlying atherogenesis, and suggest TRPM2 as a potential therapeutic target for atherosclerosis.

A Systemic Review of the Integral Role of TRPM2 in Ischemic Stroke: From Upstream Risk Factors to Ultimate Neuronal Death.

Ischemic stroke causes a heavy health burden worldwide, with over 10 million new cases every year. Despite the high prevalence and mortality rate of ischemic stroke, the underlying molecular mechanisms for the common etiological factors of ischemic stroke and ischemic stroke itself remain unclear, which results in insufficient preventive strategies and ineffective treatments for this devastating disease. In this review, we demonstrate that transient receptor potential cation channel, subfamily M, member 2 (TRPM2), a non-selective ion channel activated by oxidative stress, is actively involved in all the important steps in the etiology and pathology of ischemic stroke. TRPM2 could be a promising target in screening more effective prophylactic strategies and therapeutic medications for ischemic stroke.

Mechanism and Performance of Bituminous Mixture Using 100% Content RAP with Bio-Rejuvenated Additive (BRA).

The low RAP content, hot mixing conditions, and the addition of a high ratio of new bitumen and aggregates result in low economic and environmental benefits for current regeneration technologies. A bio-rejuvenated additive (BRA) that can fully (100%) regenerate the RAP without heating is proposed in this paper. To reveal the mechanisms of BRA-rejuvenated RAP, the effects of BRA on the chemical structure and molecular weight of the RAP were investigated using Fourier-transform infrared spectroscopy and gel permeation chromatography. The mechanical performance and water damage resistance of BRA-rejuvenated RAP were studied. Low contents of new bitumen or epoxy resin were suggested to increase the mechanical performance of 100% RAP. The results show that the 1.5% BRA-rejuvenated RAP had the best mechanical performance. The blending of BRA with recycled RAP is a completely physical process, without any chemical reactions. The molecular weight of BRA is lower than that of bitumen; it can substantially increase the content of light components in aged bitumen, and play the role of adjusting and restoring the balance of the components of aged bitumen. The mechanical performance of BRA-rejuvenated RAP is enhanced significantly by adding low dosages of new bitumen or epoxy resin.

Effects of Low-Temperature Construction Additives (LCAs) on the Performance of Asphalt Mixtures.

Green production of asphalt materials is very important to promote energy savings and emission reduction during the construction and maintenance of asphalt pavement. A low-temperature construction additive (LCA) made from the waste plastic and waste rubber is proposed, which belongs to a class of environmentally friendly additives for asphalt mixtures. Marshall stability was tested to evaluate the mechanical performance of LCA-modified asphalt mixtures (LCA-AMs). In order to determine the best preparation parameters of LCA-AMs, the influence of the content and LCA addition method on the strength of LCA-AMs was studied. In addition, the impact of epoxy resin (ER) on the mixtures' performances was evaluated. The results show that the LCA can significantly reduce the formation temperature of asphalt mixtures, and the resulting asphalt mixtures have good workability in a lower temperature range (90-110 °C). The ER should be added to the LCA-AMs after 4 h of curing. All the volumetric properties satisfy the technical requirements. The low-temperature crack resistance and fatigue resistance of LCA-AMs were obviously improved with appropriate dosages of ER, which can effectively improve the mechanical performance of the asphalt mixtures. The ER can significantly increase the rutting resistance and water sensitivity of LCA-AMs, therefore making it feasible to improve the mixture performance by the enhancement provided by a low dosage of ER.

CNNM proteins selectively bind to the TRPM7 channel to stimulate divalent cation entry into cells.

Magnesium is essential for cellular life, but how it is homeostatically controlled still remains poorly understood. Here, we report that members of CNNM family, which have been controversially implicated in both cellular Mg2+ influx and efflux, selectively bind to the TRPM7 channel to stimulate divalent cation entry into cells. Coexpression of CNNMs with the channel markedly increased uptake of divalent cations, which is prevented by an inactivating mutation to the channel's pore. Knockout (KO) of TRPM7 in cells or application of the TRPM7 channel inhibitor NS8593 also interfered with CNNM-stimulated divalent cation uptake. Conversely, KO of CNNM3 and CNNM4 in HEK-293 cells significantly reduced TRPM7-mediated divalent cation entry, without affecting TRPM7 protein expression or its cell surface levels. Furthermore, we found that cellular overexpression of phosphatases of regenerating liver (PRLs), known CNNMs binding partners, stimulated TRPM7-dependent divalent cation entry and that CNNMs were required for this activity. Whole-cell electrophysiological recordings demonstrated that deletion of CNNM3 and CNNM4 from HEK-293 cells interfered with heterologously expressed and native TRPM7 channel function. We conclude that CNNMs employ the TRPM7 channel to mediate divalent cation influx and that CNNMs also possess separate TRPM7-independent Mg2+ efflux activities that contribute to CNNMs' control of cellular Mg2+ homeostasis.

Establishing reference intervals for islet autoantibodies in Han Chinese type 1 diabetes.

Currently, islet autoantibodies (IAbs) constitute the most reliable marker for detecting the autoimmune process of type 1 diabetes (T1D). However, there are no appropriate reference intervals (RIs) to interpret the results of IAbs in China. In this study, we aimed to establish the RIs of four common IAbs based on the Han Chinese population and evaluate their clinical diagnostic values in patients with T1D. We collected 177 blood samples from healthy volunteers to detect the levels of IAbs directed against insulin (IAA), glutamic acid decarboxylase-65 (GADA), insulinoma antigen 2 (IA-2A), and zinc transporter-8 (ZnT8A) using a chemiluminescence immunoassay. RIs were calculated using nonparametric 95th percentile intervals in accordance with the Clinical and Laboratory Standards Institute guidelines, and their clinical diagnostic values were evaluated by detecting the levels of IAbs of 140 blood samples from patients with T1D in a clinical setting. We defined 138 individuals as the apparently healthy population from the 177 healthy volunteers based on the exclusion criteria. No association between the levels of the four IAbs and gender (p > .05) and age (p > .05) were found in the apparently healthy population. The combined RIs for GADA, IA-2A, ZnT8A, and IAA were 0-1.78 IU/mL, 0-3.91 IU/mL, 0-2.36 AU/mL, and 0-0.58 COI, respectively. Overall, the diagnostic efficiency for the four IAbs, especially for GADA and IAA, were improved by using the RIs established in this study. The RIs for IAbs established in this study will be a valuable tool for disease diagnosis and the therapeutic management of T1D in a clinical setting.

Substantial involvement of TRPM7 inhibition in the therapeutic effect of Ophiocordyceps sinensis on pulmonary hypertension.

Ophiocordyceps sinensis (OCS), an entomopathogenic fungus, is known to exert antiproliferative and antitissue remodeling effects. Vascular remodeling and vasoconstriction play critical roles in the development of pulmonary hypertension (PH). The therapeutic potential of OCS for PH was investigated using rodent PH models, and cultured pulmonary artery endothelial and smooth muscle cells (PAECs and PASMCs), with a focus on the involvement of TRPM7. OCS ameliorated the development of PH, right ventricular hypertrophy and dysfunction in the monocrotaline-induced PH rats. The genetic knockout of TRPM7 attenuated the development of PH in mice with monocrotaline pyrrole-induced PH. TRPM7 was associated with medial hypertrophy and the plexiform lesions in rats and humans with PH. OCS suppressed proliferation of PASMCs derived from the PH patients. Ethanol extracts of OCS inhibited TRPM7-like current, TGF-ß2-induced endothelial-mesenchymal transition, IL-6-induced STAT3 phosphorylation, and PDGF-induced Akt phosphorylation in PAECs or PASMCs. These inhibitory effects were recapitulated by either siRNA-mediated TRPM7 knockdown or treatment with TRPM7 antagonist FTY-720. OCS and FTY-720 induced vasorelaxation in the isolated normal human pulmonary artery. As a result, the present study proposes the therapeutic potential of OCS for the treatment of PH. The inhibition of TRPM7 is suggested to underlie the therapeutic effect of OCS.

Upregulation of transient receptor potential melastatin 4 (TRPM4) in ventricular fibroblasts from heart failure patients.

The transient receptor potential melastatin 4 (TRPM4) is a Ca2+-activated nonselective monovalent cation channel belonging to the TRP channel superfamily. TRPM4 is widely expressed in various tissues and most abundantly expressed in the heart. TRPM4 plays a critical role in cardiac conduction. Patients carrying a gain-of-function or loss-of-function mutation of TRPM4 display impaired cardiac conduction. Knockout or over-expression of TRPM4 in mice recapitulates conduction defects in patients. Moreover, recent studies have indicated that TRPM4 plays a role in hypertrophy and heart failure. Whereas the role of TRPM4 mediated by cardiac myocytes has been well investigated, little is known about TRPM4 and its role in cardiac fibroblasts. Here we show that in human left ventricular fibroblasts, TRPM4 exhibits typical Ca2+-activation characteristics, linear current-voltage (I-V) relation, and monovalent permeability. TRPM4 currents recorded in fibroblasts from heart failure patients (HF) are more than 2-fold bigger than those from control individuals (CTL). The enhanced functional TRPM4 in HF is not resulted from changed channel properties, as TRPM4 currents from both HF and CTL fibroblasts demonstrate similar sensitivity to intracellular calcium activation and extracellular 9-phenanthrol (9-phen) blockade. Consistent with enhanced TRPM4 activity, the protein level of TRPM4 is about 2-fold higher in HF than that of CTL hearts. Moreover, TRPM4 current in CTL fibroblasts is increased after 24 hours of TGFß1 treatment, implying that TRPM4 in vivo may be upregulated by fibrogenesis promotor TGFß1. The upregulated TRPM4 in HF fibroblasts suggests that TRPM4 may play a role in cardiac fibrogenesis under various pathological conditions.