RESUMO
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.
Assuntos
Proteínas de Transporte de Cátions/metabolismo , Ciclinas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Canais de Cátion TRPM/metabolismo , Proteínas de Transporte de Cátions/fisiologia , Cátions Bivalentes/metabolismo , Linhagem Celular Tumoral , Ciclinas/fisiologia , Células HEK293 , Humanos , Magnésio/metabolismo , Técnicas de Patch-Clamp , Proteínas Serina-Treonina Quinases/fisiologia , Canais de Cátion TRPM/genética , Canais de Cátion TRPM/fisiologiaRESUMO
Piezoelectric materials, a type of "smart" material that generates electricity while deforming and vice versa, have been used extensively for many important implantable medical devices such as sensors, transducers, and actuators. However, commonly utilized piezoelectric materials are either toxic or nondegradable. Thus, implanted devices employing these materials raise a significant concern in terms of safety issues and often require an invasive removal surgery, which can damage directly interfaced tissues/organs. Here, we present a strategy for materials processing, device assembly, and electronic integration to 1) create biodegradable and biocompatible piezoelectric PLLA [poly(l-lactic acid)] nanofibers with a highly controllable, efficient, and stable piezoelectric performance, and 2) demonstrate device applications of this nanomaterial, including a highly sensitive biodegradable pressure sensor for monitoring vital physiological pressures and a biodegradable ultrasonic transducer for blood-brain barrier opening that can be used to facilitate the delivery of drugs into the brain. These significant applications, which have not been achieved so far by conventional piezoelectric materials and bulk piezoelectric PLLA, demonstrate the PLLA nanofibers as a powerful material platform that offers a profound impact on various medical fields including drug delivery, tissue engineering, and implanted medical devices.
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Implantes Absorvíveis , Sistemas Microeletromecânicos/instrumentação , Nanofibras/química , Transdutores , Sistemas de Liberação de Medicamentos , Eletricidade , Eletrônica , Desenho de Equipamento , Monitorização Fisiológica/instrumentação , Pressão , Próteses e Implantes , Engenharia Tecidual , UltrassomRESUMO
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.
Assuntos
Fibroblastos/metabolismo , Insuficiência Cardíaca/metabolismo , Ventrículos do Coração/metabolismo , Canais de Cátion TRPM/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miócitos Cardíacos/metabolismo , Regulação para CimaRESUMO
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.
Assuntos
Diabetes Mellitus Tipo 1 , Povo Asiático , Autoanticorpos , Glutamato Descarboxilase , Humanos , InsulinaRESUMO
Measuring vital physiological pressures is important for monitoring health status, preventing the buildup of dangerous internal forces in impaired organs, and enabling novel approaches of using mechanical stimulation for tissue regeneration. Pressure sensors are often required to be implanted and directly integrated with native soft biological systems. Therefore, the devices should be flexible and at the same time biodegradable to avoid invasive removal surgery that can damage directly interfaced tissues. Despite recent achievements in degradable electronic devices, there is still a tremendous need to develop a force sensor which only relies on safe medical materials and requires no complex fabrication process to provide accurate information on important biophysiological forces. Here, we present a strategy for material processing, electromechanical analysis, device fabrication, and assessment of a piezoelectric Poly-l-lactide (PLLA) polymer to create a biodegradable, biocompatible piezoelectric force sensor, which only employs medical materials used commonly in Food and Drug Administration-approved implants, for the monitoring of biological forces. We show the sensor can precisely measure pressures in a wide range of 0-18 kPa and sustain a reliable performance for a period of 4 d in an aqueous environment. We also demonstrate this PLLA piezoelectric sensor can be implanted inside the abdominal cavity of a mouse to monitor the pressure of diaphragmatic contraction. This piezoelectric sensor offers an appealing alternative to present biodegradable electronic devices for the monitoring of intraorgan pressures. The sensor can be integrated with tissues and organs, forming self-sensing bionic systems to enable many exciting applications in regenerative medicine, drug delivery, and medical devices.
Assuntos
Implantes Absorvíveis , Monitorização Fisiológica/instrumentação , Pressão , Animais , Fenômenos Biomecânicos , Eletricidade , Humanos , Camundongos , PoliésteresRESUMO
Renin-Angiotensin-Aldosterone System (RAAS) measurements are influenced by several factors. We investigated the effect of sample delivery conditions on RAAS measurements including sample storage temperature and time. Blood samples were collected from thirty participants using enzyme inhibitor tubes and serum separation gel evacuated tubes. Plasma and serum from fresh blood samples without further storage (as baseline), and from blood samples that were stored at either 0 °C, 4 °C, or 25 °C for 3 h, 6 h and 24 h, respectively, were extracted and stored at -30 °C for batch measurements using radioimmunoassay. Concentrations of Aldosterone (Ald) decreased following delivery temperature and time, and were significantly different when samples were set aside at 0 °C for 24 h (p < .01), 4 °C for 6 h (p < .01), and 25 °C for 3 h (p < .05). However, levels of Angiotensin (Ang I) increased following delivery temperature and time, and were significantly different when samples were set aside at 0 °C and 4 °C for 6 h (p < .05) and at 25 °C for 3 h (p < .001). However, no changes were observed for the concentrations of plasma renin activity (PRA) and Ang II, except for Ang II which increased significantly when samples were set aside at 25 °C for 24 h (p < .001). Our results indicate that samples used for RAAS measurement should be placed at a low temperature and analyzed as soon as possible after collection.
Assuntos
Aldosterona/sangue , Angiotensina II/sangue , Angiotensina I/sangue , Radioimunoensaio/normas , Renina/sangue , Manejo de Espécimes/normas , Adulto , Idoso , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Refrigeração/normas , Sistema Renina-Angiotensina/genéticaRESUMO
The atrial-specific ultrarapid delayed rectifier K+ current (IKur) inactivates slowly but completely at depolarized voltages. The consequences for IKur rate-dependence have not been analyzed in detail and currently available mathematical action-potential (AP) models do not take into account experimentally observed IKur inactivation dynamics. Here, we developed an updated formulation of IKur inactivation that accurately reproduces time-, voltage-, and frequency-dependent inactivation. We then modified the human atrial cardiomyocyte Courtemanche AP model to incorporate realistic IKur inactivation properties. Despite markedly different inactivation dynamics, there was no difference in AP parameters across a wide range of stimulation frequencies between the original and updated models. Using the updated model, we showed that, under physiological stimulation conditions, IKur does not inactivate significantly even at high atrial rates because the transmembrane potential spends little time at voltages associated with inactivation. Thus, channel dynamics are determined principally by activation kinetics. IKur magnitude decreases at higher rates because of AP changes that reduce IKur activation. Nevertheless, the relative contribution of IKur to AP repolarization increases at higher frequencies because of reduced activation of the rapid delayed-rectifier current IKr. Consequently, IKur block produces dose-dependent termination of simulated atrial fibrillation (AF) in the absence of AF-induced electrical remodeling. The inclusion of AF-related ionic remodeling stabilizes simulated AF and greatly reduces the predicted antiarrhythmic efficacy of IKur block. Our results explain a range of experimental observations, including recently reported positive rate-dependent IKur-blocking effects on human atrial APs, and provide insights relevant to the potential value of IKur as an antiarrhythmic target for the treatment of AF.
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Fibrilação Atrial/metabolismo , Átrios do Coração/metabolismo , Canais de Potássio/metabolismo , Antiarrítmicos/farmacologia , Fibrilação Atrial/tratamento farmacológico , Relação Dose-Resposta a Droga , Átrios do Coração/efeitos dos fármacos , Humanos , Cinética , Potenciais da Membrana/efeitos dos fármacos , Potenciais da Membrana/fisiologia , Modelos Cardiovasculares , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , Bloqueadores dos Canais de Potássio/farmacologiaRESUMO
Organophosphorous nerve agents (NAs) pose a great threat to nations and people because of their acute and extreme toxicities. The rapid detection of NAs has attracted growing interest in the first emergency response field. In this work, we demonstrate a simple and sensitive surface-enhanced Raman spectroscopic (SERS) method for NAs detection, and G- and V-agents discrimination. The results show that VX (V-agents) can be directly detected at a 20 ng mL-1 level with pinhole shell-isolated gold nanoparticles (pinSHINs) as the substrate. Moreover, combined with a specific and prompt alkaline keto-oxime transformation approach in a full aqueous solution, G-agents can be measured as low as 10 ng mL-1 with excellent discrimination from V-agents and other common organophosphorous pesticides within several minutes. The achieved discriminative detection of G-agents and VX could be significant not only for reducing the false positive and negative signals but also for providing an appropriate recommendation on the effective medical rescue. A decontamination outcome occurred alongside, any highly toxic G-agents were converted to a less toxic phosphate, and the generated cyanide was fully and firmly adsorbed onto the surface of pinSHINs substrate, which may be further used for on-site detection of extremely toxic NA prototypes. Graphical abstract á .
Assuntos
Agentes Neurotóxicos/análise , Compostos Organofosforados/análise , Análise Espectral Raman/métodos , Limite de Detecção , Microscopia Eletrônica de Transmissão , Propriedades de SuperfícieRESUMO
To identify novel targets for neuropathic pain, 3097 mouse knockout lines were tested in acute and persistent pain behavior assays. One of the lines from this screen, which contained a null allele of the adapter protein-2 associated kinase 1 (AAK1) gene, had a normal response in acute pain assays (hot plate, phase I formalin), but a markedly reduced response to persistent pain in phase II formalin. AAK1 knockout mice also failed to develop tactile allodynia following the Chung procedure of spinal nerve ligation (SNL). Based on these findings, potent, small-molecule inhibitors of AAK1 were identified. Studies in mice showed that one such inhibitor, LP-935509, caused a reduced pain response in phase II formalin and reversed fully established pain behavior following the SNL procedure. Further studies showed that the inhibitor also reduced evoked pain responses in the rat chronic constriction injury (CCI) model and the rat streptozotocin model of diabetic peripheral neuropathy. Using a nonbrain-penetrant AAK1 inhibitor and local administration of an AAK1 inhibitor, the relevant pool of AAK1 for antineuropathic action was found to be in the spinal cord. Consistent with these results, AAK1 inhibitors dose-dependently reduced the increased spontaneous neural activity in the spinal cord caused by CCI and blocked the development of windup induced by repeated electrical stimulation of the paw. The mechanism of AAK1 antinociception was further investigated with inhibitors of α2 adrenergic and opioid receptors. These studies showed that α2 adrenergic receptor inhibitors, but not opioid receptor inhibitors, not only prevented AAK1 inhibitor antineuropathic action in behavioral assays, but also blocked the AAK1 inhibitor-induced reduction in spinal neural activity in the rat CCI model. Hence, AAK1 inhibitors are a novel therapeutic approach to neuropathic pain with activity in animal models that is mechanistically linked (behaviorally and electrophysiologically) to α2 adrenergic signaling, a pathway known to be antinociceptive in humans.
Assuntos
Neuralgia/tratamento farmacológico , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Animais , Fenômenos Eletrofisiológicos/efeitos dos fármacos , Técnicas de Inativação de Genes , Células HEK293 , Humanos , Masculino , Camundongos , Neuralgia/metabolismo , Neuralgia/fisiopatologia , Nociceptividade/efeitos dos fármacos , Fenótipo , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Ratos , Medula Espinal/efeitos dos fármacos , Medula Espinal/enzimologia , Medula Espinal/fisiopatologiaRESUMO
A highly sensitive method for the determination of sulfur mustard (SM) metabolites thiodiglycol (TDG) and thiodiglycol sulfoxide (TDGO) in urine was established and validated using isotope-dilution negative-ion chemical ionization (NICI) gas chromatography-mass spectrometry (GC-MS). TDGO in the samples was reduced with TiCl3, and then determined together with TDG as a single analyte. The sample preparation procedures, including two solid-phase-extraction (SPE) clean-up steps, were optimized to improve the sensitivity of the method. The limits of detection (LOD) for both TDG and TDG plus TDGO (TDG + TDGO) were 0.1 ng mL(-1), and the limits of quantitation (LOQ) for both were 0.3 ng mL(-1). The method was used in a rabbit cutaneous SM exposure model. Domestic rabbits were exposed to neat liquid SM at three dosage levels (0.02, 0.05, and 0.15 LD50), and the urinary excretion of four species of hydrolysis metabolites, namely free TDG, free plus conjugated TDG (total TDG), free TDG + TDGO, and free plus conjugated TDG + TDGO (total TDG + TDGO), was evaluated to investigate the metabolic processes. The total urinary excretion profiles of the metabolites, including the peak time, time window, and dose-response and time-response relationships, were clarified. The results revealed that the concentrations of TDG and TDG + TDGO in the urine increased quickly and then decreased rapidly in the first two days after SM exposure. The cumulative amount of total TDG + TDGO excreted in urine during the first five days accounted for 0.5-1% of the applied dose of SM. It is also concluded that TDG and TDGO in urine existed mainly in free form, the levels of glucuronide and of sulfate conjugates of TDG or TDGO were very low, and most hydrolysis metabolites were present in the oxidized form (TDGO). The study indicates that the abnormal increase of TDG and TDGO excretion levels can be used as a diagnostic indicator and establishes a reference time-window for retrospective analysis and sampling after SM exposure.
Assuntos
Substâncias para a Guerra Química/toxicidade , Fármacos Dermatológicos/toxicidade , Gás de Mostarda/toxicidade , Compostos de Sulfidrila/urina , Sulfóxidos/urina , Administração Cutânea , Animais , Biotransformação , Substâncias para a Guerra Química/metabolismo , Fármacos Dermatológicos/metabolismo , Relação Dose-Resposta a Droga , Cromatografia Gasosa-Espectrometria de Massas/métodos , Técnicas de Diluição do Indicador , Masculino , Gás de Mostarda/metabolismo , Oxirredução , Coelhos , Pele/irrigação sanguínea , Pele/efeitos dos fármacos , Pele/metabolismo , Extração em Fase Sólida , Titânio/químicaRESUMO
A simple and sensitive method has been developed and validated for determining ethyl methylphosphonic acid (EMPA), isopropyl methylphosphonic acid (IMPA), isobutyl methylphosphonic acid (iBuMPA), and pinacolyl methylphosphonic acid (PMPA) in human urine using gas chromatography-tandem mass spectrometry (GC-MS/MS) coupled with solid phase derivatization (SPD). These four alkyl methylphosphonic acids (AMPAs) are specific hydrolysis products and biomarkers of exposure to classic organophosphorus (OP) nerve agents VX, sarin, RVX, and soman. The AMPAs in urine samples were directly derivatized with pentafluorobenzyl bromide on a solid support and then extracted by liquid-liquid extraction. The analytes were quantified with isotope-dilution by negative chemical ionization (NCI) GC-MS/MS in a selected reaction monitoring (SRM) mode. This method is highly sensitive, with the limits of detection of 0.02 ng/mL for each compound in a 0.2 mL sample of human urine, and an excellent linearity from 0.1 to 50 ng/mL. It is proven to be very suitable for the qualitative and quantitative analyses of degradation markers of OP nerve agents in biomedical samples.
Assuntos
Substâncias para a Guerra Química/análise , Organofosfonatos/urina , Compostos Organofosforados/urina , Compostos Organotiofosforados/urina , Sarina/urina , Soman/análogos & derivados , Soman/urina , Biotransformação , Substâncias para a Guerra Química/metabolismo , Fluorbenzenos/química , Cromatografia Gasosa-Espectrometria de Massas/métodos , Humanos , Técnicas In Vitro , Técnicas de Diluição do Indicador , Limite de Detecção , Extração Líquido-LíquidoRESUMO
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.
RESUMO
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.
Assuntos
Lesões Encefálicas , Isquemia Encefálica , AVC Isquêmico , Acidente Vascular Cerebral , Canais de Cátion TRPM , Humanos , Camundongos , Animais , Barreira Hematoencefálica/metabolismo , AVC Isquêmico/metabolismo , Células Endoteliais/metabolismo , Canais de Cátion TRPM/metabolismo , Cálcio/metabolismo , Células HEK293 , Oxigênio , Lesões Encefálicas/metabolismo , Acidente Vascular Cerebral/metabolismo , Isquemia Encefálica/metabolismoRESUMO
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.
Assuntos
Lesões Encefálicas , Canais de Cátion TRPM , Humanos , Proteínas Quinases/metabolismo , Canais de Cátion TRPM/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Peptídeos/metabolismoRESUMO
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.
RESUMO
PURPOSE: The purpose of this study was to evaluate left ventricular (LV) mechanical dyssynchrony in patients with Wolff-Parkinson-White (WPW) syndrome pre- and post-radiofrequency catheter ablation (RFA) using phase analysis of gated single photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI). METHODS: Forty-five WPW patients were enrolled and had gated SPECT MPI pre- and 2-3 days post-RFA. Electrophysiological study (EPS) was used to locate accessory pathways (APs) and categorize the patients according to the AP locations (septal, left and right free wall). Electrocardiography (ECG) was performed pre- and post-RFA to confirm successful elimination of the APs. Phase analysis of gated SPECT MPI was used to assess LV dyssynchrony pre- and post-RFA. RESULTS: Among the 45 patients, 3 had gating errors, and thus 42 had SPECT phase analysis. Twenty-two patients (52.4%) had baseline LV dyssynchrony. Baseline LV dyssynchrony was more prominent in the patients with septal APs than in the patients with left or right APs (p < 0.05). RFA improved LV synchrony in the entire cohort and in the patients with septal APs (p < 0.01). CONCLUSION: Phase analysis of gated SPECT MPI demonstrated that LV mechanical dyssynchrony can be present in patients with WPW syndrome. Septal APs result in the greatest degree of LV mechanical dyssynchrony and afford the most benefit after RFA. This study supports further investigation in the relationship between electrical and mechanical activation using EPS and phase analysis of gated SPECT MPI.
Assuntos
Feixe Acessório Atrioventricular/diagnóstico por imagem , Tecnécio Tc 99m Sestamibi , Disfunção Ventricular Esquerda/diagnóstico por imagem , Síndrome de Wolff-Parkinson-White/diagnóstico por imagem , Feixe Acessório Atrioventricular/complicações , Feixe Acessório Atrioventricular/patologia , Adulto , Tomografia Computadorizada por Emissão de Fóton Único de Sincronização Cardíaca/métodos , Ablação por Cateter/métodos , Ecocardiografia/métodos , Eletrocardiografia/métodos , Técnicas Eletrofisiológicas Cardíacas/métodos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Imagem de Perfusão do Miocárdio/métodos , Compostos Radiofarmacêuticos , Disfunção Ventricular Esquerda/etiologia , Disfunção Ventricular Esquerda/patologia , Síndrome de Wolff-Parkinson-White/complicações , Síndrome de Wolff-Parkinson-White/patologiaRESUMO
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.
Assuntos
AVC Isquêmico , Canais de Cátion TRPM , Humanos , Morte Celular , Estresse Oxidativo , Fatores de Risco , Canais de Cátion TRPM/genética , Canais de Cátion TRPM/metabolismoRESUMO
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.
RESUMO
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.
RESUMO
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.