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1.
J Agric Food Chem ; 2020 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-32396372

RESUMO

Inoculation of crop plants with strains of beneficial bacteria can result in promotion of plant growth. In our study, we demonstrated that Bacillus velezensis WRN031 as a plant-growth-promoting rhizobacteria (PGPR) improved the maize seedling growth following inoculation with WRN031. Fluorescence microscopy visualization indicated that GFP-labeled B. velezensis WRN031 had accumulated on the maturation zones of both primary and lateral roots of maize. Two metabolites were detected in the rhizosphere soil of maize root inoculation with WRN031 using HPLC-DAD analyses. Through guided isolation from an ethyl acetate extract of B. velezensis WRN031, these two nonvolatile meso stereoisomers 3S,4R-acetylbutanediol (3S,4R-ABD, 1) and 3R,4R-acetylbutanediol (3R,4R-ABD, 2) were identified and found to occur at a ratio of 1:2 (v/v) in maize rhizosphere soil. Bioactivity screening indicated that compounds 1 and 2, as well as a v/v = 1:2 mixture of both 1 and 2, significantly improved the root elongation of both maize and rice, with the effective enhancement concentration related to their concentration in rhizosphere soil. These results suggested that 3S,4R-ABD and 3R,4R-ABD produced by B. velezensis WRN031 might improve the growth of their host plants and provides evidence that nonvolatiles accumulating in the root maturation zone may regulate the relationship between roots and beneficial bacteria.

2.
Thorac Cancer ; 2020 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-32433828

RESUMO

BACKGROUND: Although many studies have defined mechanisms of resistance to EGFR-TKIs, acquired resistance remains the major limitation of monotherapy with EGFR-TKIs. METHODS: Cell viability was analyzed using a Cell Counting Kit-8 (CCK-8) assay. EGFR T790M mutation was sequenced on a HiSeq 4000 platform. mRNAs from HCC827 and HCC827 gefitinib-resistant (GR) cells were analyzed by genome analyzer-based deep sequencing. The effect of anlotinib on apoptosis and cell cycle arrest of HCC827 GR was detected by fluorescence-activated cell sorting (FACS) analysis. A mouse xenograft model was used to assess the effect of anlotinib on HCC827 GR cells. RESULTS: The T790M mutation was found in the PC-9 GR cell line but not in the HCC827 GR cell line. Anlotinib could suppress the growth of HCC827 GR cells by inhibiting FGFR1 in vitro and in a mouse xenograft model. Moreover, FGFR1 was overexpressed in HCC827 GR cells, and the knockdown of FGFR1 reversed gefitinib resistance in HCC827 GR cells. Furthermore, anlotinib induced apoptosis and cell cycle arrest in HCC827 GR cells by increasing the activity of Caspase-3. CONCLUSIONS: FGFR1 overexpression could be the mechanism of EGFR-TKI acquired resistance and anlotinib can suppresse the growth of EGFR-TKI-resistant NSCLC cells without T790M mutation.

3.
Stem Cell Res ; 45: 101832, 2020 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-32416579

RESUMO

Mitochondrial DNA depletion syndrome-13 (MTDPS13) is a rare autosomal recessive mitochondrial disease caused by mutations in the FBXL4 (F-box and leucine-rich repeat protein 4) gene. Using Sendai virus-mediated reprogramming, we established an induced pluripotent stem cell (iPSC) line from PBMCs collected from a one-year-old female patient with MTDPS13. The iPSCs were stable during amplification, expressed pluripotent genes, maintained a normal karyotype, and showed characteristics of the three germs layers in an in vitro differentiation assay.

4.
Genes Genomics ; 2020 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-32462516

RESUMO

BACKGROUND: Hepatitis B virus (HBV) infection is a crucial medical issue worldwide. The dependence of HBV replication on host cell machineries and their co-evolutionary interactions prompt the codon usage pattern of viral genes to translation selection and mutation pressure. OBJECTIVE: The evolutionary characteristics of HBV and the natural selection effects of the human genome on the codon usage characteristics were analyzed to provide a basis for medication development for HBV infection. METHODS: The codon usage pattern of sequences from different HBV genotypes of our isolates and reference HBV genome sequences downloaded from the National Center for Biotechnology Information (NCBI) database were analyzed by computing the relative synonymous codon usage (RSCU), nucleotide content, codon adaptation index (CAI) and the effective number of codons (ENC). RESULTS: The highest ENC values were observed in the C genotypes, followed by the B genotypes. The ENC values indicated a weak codon usage bias (CUB) in HBV genome. The number of codons differentially used between the three genotypes was markedly higher than that of similarly used codons. High CAI values indicated a good adaptability of HBV to its host. The ENC plot indicated the occurrence of mutational pressure in the three genotypes. The mean Ka/Ks ratios in the three genotypes were lower than 1, which indicated a negative selection pressure. The CAI and GC3% plot indicated the existence of CUB in the HBV genome. CONCLUSIONS: Nucleotide composition, mutation bias, negative selection and mutational pressure are key factors influencing the CUB and phylogenetic diversity in HBV genotypes. The data provided here could be useful for developing drugs for HBV infection.

5.
Phys Rev Lett ; 124(15): 156601, 2020 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-32357024

RESUMO

The notion of topological phases has been extended to higher-order and has been generalized to different dimensions. As a paradigm, Cd_{3}As_{2} is predicted to be a higher-order topological semimetal, possessing three-dimensional bulk Dirac fermions, two-dimensional Fermi arcs, and one-dimensional hinge states. These topological states have different characteristic length scales in electronic transport, allowing one to distinguish their properties when changing sample size. Here, we report an anomalous dimensional reduction of supercurrent transport by increasing the size of Dirac semimetal Cd_{3}As_{2}-based Josephson junctions. An evolution of the supercurrent quantum interferences from a standard Fraunhofer pattern to a superconducting quantum interference device (SQUID)-like one is observed when the junction channel length is increased. The SQUID-like interference pattern indicates the supercurrent flowing through the 1D hinges. The identification of 1D hinge states should be valuable for deeper understanding of the higher-order topological phase in a 3D Dirac semimetal.

6.
Pest Manag Sci ; 2020 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-32246522

RESUMO

BACKGROUND: The development of efficient and safe green pesticides is a scientific strategy to alleviate current pesticide residues, environmental pollution, and threats to non-target organisms. Pesticide controlled-release formulations (CRFs) have attracted wide attention because they can control the rate of release of active ingredients and prolong the effective duration. In particular, nanoscale pesticide sustained-release systems have excellent biological activity and distribution performance because of their small particle size. Some technical difficulties remain in obtaining nanoscale CRFs. RESULTS: We successfully fabricated pyraclostrobin nanosphere CRF by combining high-pressure homogenization technology and emulsion-solvent evaporation methods. The pyraclostrobin nanospheres had a uniform spherical shape with a mean particle size of 450 nm and polydispersity index of less than 0.3. The pyraclostrobin loading capacity reached 53.6%, with excellent storage stability. The contact angle of nanospheres on cucumber leaf surfaces demonstrated that it had good wettability. Compared with pyraclostrobin technical and commercial formulations, the nanosphere systems showed a significantly sustained release of pyraclostrobin for longer (up to 250 h). A preliminary bioassay against Penicillium ochrochloron showed that the bioactivity and long-term efficiency of pyraclostrobin nanospheres were superior to those of the commercial formulation. CONCLUSION: This research introduced a simple, fast, expandable method for preparing pyraclostrobin nanospheres. The results showed that pyraclostrobin nanospheres could prolong the duration of pesticide efficacy and enhance bioactivity. Furthermore, this technology provides a platform for scale-up production of nano-scale pesticide CRFs.

7.
Phys Rev Lett ; 124(11): 116802, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32242698

RESUMO

The exotic topological surface states of Dirac or Weyl semimetals, namely Fermi arcs, are predicted to be spin polarized, while their spin polarization nature is still not revealed by transport measurements. Here, we report the spin-polarized transport in a Dirac semimetal Cd_{3}As_{2} nanowire employing the ferromagnetic electrodes for spin detection. The spin-up and spin-down states can be changed by reversing the current polarity, showing the spin-momentum locking property. Moreover, the nonlocal measurements show a high fidelity of the spin signals, indicating the topological protection nature of the spin transport. As tuning the Fermi level away from the Dirac point by gate voltages, the spin signals gradually decrease and finally are turned off, which is consistent with the fact that the Fermi arc surface state has the maximum ratio near the Dirac point and disappears above the Lifshitz transition point. Our results should be valuable for revealing the transport properties of the spin-polarized Fermi arc surface states in topological semimetals.

8.
ACS Nano ; 2020 Apr 14.
Artigo em Inglês | MEDLINE | ID: mdl-32286783

RESUMO

Characterized by bulk Dirac or Weyl cones and surface Fermi-arc states, topological semimetals have sparked enormous research interest in recent years. The nanostructures, with large surface-to-volume ratio and easy field-effect gating, provide ideal platforms to detect and manipulate the topological quantum states. Exotic physical properties originating from these topological states endow topological semimetals attractive for future topological electronics (topotronics). For example, the linear energy dispersion relation is promising for broadband infrared photodetectors, the spin-momentum locking nature of topological surface states is valuable for spintronics, and the topological superconductivity is highly desirable for fault-tolerant qubits. For real-life applications, topological semimetals in the form of nanostructures are necessary in terms of convenient fabrication and integration. Here, we review the recent progresses in topological semimetal nanostructures and start with the quantum transport properties. Then topological semimetal-based electronic devices are introduced. Finally, we discuss several important aspects that should receive great effort in the future, including controllable synthesis, manipulation of quantum states, topological field effect transistors, spintronic applications, and topological quantum computation.

9.
Zhongguo Zhen Jiu ; 40(3): 243-6, 2020 Mar 12.
Artigo em Chinês | MEDLINE | ID: mdl-32270634

RESUMO

OBJECTIVE: To observe the effect of transcutaneous electrical acupoint stimulation (TEAS) on postoperative olfactory memory disorder in patients with general anesthesia of sevoflurane and to explore its possible mechanism. METHODS: Forty patients who were scheduled to have gynecological and urological procedures under general anesthesia were randomly divided into an observation group and a control group, 20 cases in each group. The patients in the observation group were treated with TEAS (dilatational wave, 2 Hz/100 Hz) at Yingxiang (LI 20) and Yintang (GV 29) 10 min before anesthesia induction until the end of operation; the patients in the control group received general anesthesia directly. The changes of mean arterial pressure (MAP), heart rate (HR) and blood oxygen saturation (SpO2) were recorded before treatment, 30 min after operation and at the end of operation; smell identification score was measured by Connecticut Chemosensory Clinical Research Center (CCCRC) olfactory test before treatment (T0) and when Aldrete recovery score reached 10 points at the end of anesthesia (T1); the concentration of melatonin in plasma was measured by ELISA method in the two groups. RESULTS: The between-group differences and within-group differences of MAP, HR and SpO2 were not significant at each time point (P>0.05). Compared with T0, the score of smell identification and plasma concentration of melatonin were not significantly different at T1 in the observation group (P>0.05), however, the score of smell identification and plasma concentration of melatonin were reduced in the control group (P<0.05). At T1, the score of smell identification and plasma concentration of melatonin in the observation group were higher than those in the control group (P<0.05). CONCLUSION: TEAS could improve the postoperative olfactory memory disorder in patients with general anesthesia of sevoflurane, and its mechanism may be related to the increase of plasma concentration of melatonin.

10.
Int J Environ Health Res ; : 1-10, 2020 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-32281876

RESUMO

To assess the association between fluoride exposure and children's behavioural outcomes, we recruited 325 resident school-age children (7-13 years old) lived in Tongxu County of Henan Province in China. We measured urinary fluoride (UF) concentrations using the ion-selective electrode method. Children's behavioural outcomes were assessed by Conners' Parent Rating Scale-Revised, including conduct problems, learning problems, psychosomatic problems, impulsive-hyperactive, anxiety, and ADHD index. It turned out that each 1.0 mg/L increment in UF concentration corresponded with an elevation in the psychosomatic problem score of 4.01 (95% CI: 2.74, 5.28) and a 97% (OR = 1.97, 95% CI: 1.19, 3.27) increase in the prevalence of psychosomatic problems after adjusting for potential influencing factors. The sensitivity analysis results were consistent with those observed in our preliminary analysis. Our study suggests that fluoride exposure is positively related to the behavioural problem in school-age children, psychosomatic problem in particular.

11.
Nature ; 579(7798): 224-228, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32123353

RESUMO

Large-scale energy storage is becoming increasingly critical to balancing renewable energy production and consumption1. Organic redox flow batteries, made from inexpensive and sustainable redox-active materials, are promising storage technologies that are cheaper and less environmentally hazardous than vanadium-based batteries, but they have shorter lifetimes and lower energy density2,3. Thus, fundamental insight at the molecular level is required to improve performance4,5. Here we report two in situ nuclear magnetic resonance (NMR) methods of studying redox flow batteries, which are applied to two redox-active electrolytes: 2,6-dihydroxyanthraquinone (DHAQ) and 4,4'-((9,10-anthraquinone-2,6-diyl)dioxy) dibutyrate (DBEAQ). In the first method, we monitor the changes in the 1H NMR shift of the liquid electrolyte as it flows out of the electrochemical cell. In the second method, we observe the changes that occur simultaneously in the positive and negative electrodes in the full electrochemical cell. Using the bulk magnetization changes (observed via the 1H NMR shift of the water resonance) and the line broadening of the 1H shifts of the quinone resonances as a function of the state of charge, we measure the potential differences of the two single-electron couples, identify and quantify the rate of electron transfer between the reduced and oxidized species, and determine the extent of electron delocalization of the unpaired spins over the radical anions. These NMR techniques enable electrolyte decomposition and battery self-discharge to be explored in real time, and show that DHAQ is decomposed electrochemically via a reaction that can be minimized by limiting the voltage used on charging. We foresee applications of these NMR methods in understanding a wide range of redox processes in flow and other electrochemical systems.


Assuntos
Fontes de Energia Elétrica , Espectroscopia de Ressonância Magnética , Eletrólitos/química , Elétrons , Oxirredução
12.
Food Chem ; 319: 126568, 2020 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-32169768

RESUMO

Penthorum chinense Pursh is a dietary medicinal plant widely distributed in Asia-Pacific countries. The present study aims to profile the chemical constituents of P. chinense and investigate its prebiotic role in modulating gut microbiota. Fifty polyphenolic compounds were rapidly identified using UPLC-HR-MS. Total flavonoid and phenolic contents of P. chinense were 46.6% and 61.3% (w/w), respectively. Thirteen individual polyphenols were quantified, which accounted for 33.1% (w/w). P. chinense induced structural arrangement of microbial community in mice, showing increased microbiota diversity, elevated Bacteroidetes/Firmicutes ratio and enriched gut health-promoting bacteria. After a one-week drug-free wash, most of these changes were recovered, but the abundance of some beneficial bacteria was further increased. The altered composition of gut microbiota enriched several metabolic pathways. Moreover, P. chinense increased antioxidant capacity in vivo. The results suggest that polyphenol-enriched P. chinense modulates gut microbiota and enhances antioxidant capacity in mice toward a beneficial environment for host health.

13.
Artigo em Inglês | MEDLINE | ID: mdl-32133738

RESUMO

Membranes which allow fast and selective transport of protons and cations are required for a wide range of electrochemical energy conversion and storage devices, such as proton-exchange membrane (PEM) fuel cells (PEMFCs) and redox flow batteries (RFBs). Herein we report a new approach to designing solution-processable ion-selective polymer membranes with both intrinsic microporosity and ion-conductive functionality. Polymers are synthesized with rigid and contorted backbones, which incorporate hydrophobic fluorinated and hydrophilic sulfonic acid functional groups, to produce membranes with negatively charged subnanometer-sized confined ionic channels. The ready transport of protons and cations through these membranes, and the high selectivity towards nanometer-sized redox-active molecules, enable efficient and stable operation of an aqueous alkaline quinone redox flow battery and a hydrogen PEM fuel cell.

14.
Artigo em Inglês | MEDLINE | ID: mdl-32212243

RESUMO

The advent of immune checkpoint inhibitors has improved survival in some types of cancer and brought promising prospects to cancer immunotherapy. Despite their clinical benefits, significant off-target toxicities resulting from the immune system activation have been observed, namely immune-related adverse events (irAEs), which pose to clinicians a new challenge of optimal management. With steroids being the mainstay of current management of irAEs, immunosuppressive agents are especially indicated for severe or steroid-refractory cases, based on current immunopathophysiological knowledge and on extrapolations of treatment options for primary autoimmune disorders. This review focuses on the status and recent clinical progress of immunosuppressive agents in the management of severe and steroid-refractory irAEs.

15.
Metab Brain Dis ; 35(4): 627-635, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32140823

RESUMO

The study found that microRNAs play an important role in Parkinson's disease (PD). However, the function of MicroRNA-216a (miR-216a) in PD is unclear. Therefore, this experiment aimed to investigate the pathogenesis of miR-216a in PD. Using the toxicity of MPP+ to polyhexamine neurons, apoptosis of SH-SY5Y neuroblastoma cells was induced at different time by MPP+ to construct a stable acute PD cell model. The effects of DNA breakage, mitochondrial membrane potential (A ^ m), caspase-3 activity and nucleosome enrichment on cell apoptosis were detected by flow cytometry, TUNEL. MPP+ increased the toxic effects of dopaminergic neurons in a PD model. The introduction of miR-216a inhibited MPP + -induced neuronal apoptosis. The main manifestations were the decreased levels of positive rate of Tunel cells, caspase 3 activity and nucleosome enrichment factor. Bax was a direct target of miR-216a. In addition, Bax overexpression reversed the effects of miR-216a on neural cells. Bax downstream factors were also involved in miR-216a regulation of MPP + -triggered neuronal apoptosis. miR-216a regulated the progression of PD by regulating Bax, and miR-216a may be a potential target for PD.

16.
Nutr Neurosci ; : 1-10, 2020 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-31973664

RESUMO

Aim: To measure the development of moderate to late preterm children by Ages and Stages Questionnaires (ASQ) and explore the relationship between moderate to late preterm, diet types and development delay in less-developed rural China.Methods: Data were collected from a cross-sectional community-based survey, which recruited 1748 children aged 1-59 months in eight counties of China. Caregivers of these children completed the Chinese version of ASQ-3 (ASQ-C) while physical examination and questionnaires on socio-demographic characteristics were conducted. Multivariate logistic regressions were used to analyze the association between moderate to late preterm and suspected developmental delay, as well as the association between diet types and suspected developmental delay. Consumption of certain food types was compared between moderate to late preterm and full-term children.Results: The prevalence of suspected overall developmental delay was 31.3% in the moderate to the late preterm group, compared with 21.6% in the full-term group. Moderate to late preterm birth was not associated with total suspected developmental delay and developmental delay in all the domains of ASQ, except for fine motor (OR = 2.43 95% C.I.: 1.04-5.56). The intake of vegetables and fruits had a protective influence on developmental delay in fine motor function, and moderate to late preterm children had lower relative consumption of fruits and vegetables than full-term children.Conclusion: Moderate to late preterm children in rural China showed an increased likelihood of developmental delay in fine motor function. Future interventions to improve the intake of vegetables and fruits in moderate to late preterm children are recommended.

17.
Dev Cell ; 52(3): 309-320.e5, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-31902656

RESUMO

Movement of chromosome sites within interphase cells is critical for numerous pathways including RNA transcription and genome organization. Yet, a mechanism for reorganizing chromatin in response to these events had not been reported. Here, we delineate a molecular chaperone-dependent pathway for relocating activated gene loci in yeast. Our presented data support a model in which a two-authentication system mobilizes a gene promoter through a dynamic network of polymeric nuclear actin. Transcription factor-dependent nucleation of a myosin motor propels the gene locus through the actin matrix, and fidelity of the actin association was ensured by ARP-containing chromatin remodelers. Motor activity of nuclear myosin was dependent on the Hsp90 chaperone. Hsp90 further contributed by biasing the remodeler-actin interaction toward nucleosomes with the non-canonical histone H2A.Z, thereby focusing the pathway on select sites such as transcriptionally active genes. Together, the system provides a rapid and effective means to broadly yet selectively mobilize chromatin sites.

18.
J Ethnopharmacol ; 251: 112569, 2020 Apr 06.
Artigo em Inglês | MEDLINE | ID: mdl-31935496

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: In China, Penthorum chinense Pursh (P. chinense) has been used for hundreds of years traditionally for alleviating symptoms by excessive intake of alcohol as well as in the treatment of traumatic injury, edema and liver diseases. Recently, P. chinense and its extract have been developed into tea, drinks or medicines for treatment of liver diseases, including hepatic virus infections, alcoholic liver disease (ALD), non-alcoholic fatty liver disease (NAFLD) and liver fibrosis. AIM OF THE STUDY: The main purpose of this review is to provide a critical appraisal of the existing knowledge on the phytochemical data, quality control aspect, pharmacological, as well as toxicological and clinical studies performed on P. chinense, including the identification of scientific gaps. MATERIALS AND METHODS: A detailed literature search was conducted using various online search engines, such as Pubmed, Scopus, Google Scholar, Mendeley, Web of Science as well as China National Knowledge Infrastructure (CNKI) database. RESULTS: In the pharmacological studies, there clearly are links between local/traditional uses and the biomedical investigations. Most pharmacological studies indicated potential liver protective effects in experimental models of chemicals-induced liver injury, acute and chronic alcoholic liver injury, NAFLD, liver fibrosis and viral infection, potentially through antioxidant effects, balancing key liver enzyme levels, inhibition of hepatic virus DNA replication, inhibition of hepatic stellate cells activation and inflammation either in vitro or in vivo. In some models, the effects of P. chinense is comparable with the one of silymarin. Clinical studies have suggested that P. chinense is safe and effective in treating several liver diseases, although most of them are not double-blinded and placebo-controlled studies. Toxicology studies show that P. chinense has no obvious toxicity or side effects in animals or human. Flavonoids, lignans, coumarins, polyphenols and organic acids have been identified. However, only a few studies have investigated the active compounds (mainly flavonoids and lignans) and molecular mechanisms of P. chinense. CONCLUSION: P. chinense seems to be safe and shows relevant liver protecting effects. Therefore, it might be a promising candidate for developing as new hepatoprotective agents. However, a lack of understanding of the active compounds and mechanisms of action needs further attention.

19.
Genome Biol ; 21(1): 14, 2020 01 17.
Artigo em Inglês | MEDLINE | ID: mdl-31952552

RESUMO

The error-prone third-generation sequencing (TGS) long reads can be corrected by the high-quality second-generation sequencing (SGS) short reads, which is referred to as hybrid error correction. We here investigate the influences of the principal algorithmic factors of two major types of hybrid error correction methods by mathematical modeling and analysis on both simulated and real data. Our study reveals the distribution of accuracy gain with respect to the original long read error rate. We also demonstrate that the original error rate of 19% is the limit for perfect correction, beyond which long reads are too error-prone to be corrected by these methods.

20.
Nat Mater ; 19(2): 195-202, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31792424

RESUMO

Membranes with fast and selective ion transport are widely used for water purification and devices for energy conversion and storage including fuel cells, redox flow batteries and electrochemical reactors. However, it remains challenging to design cost-effective, easily processed ion-conductive membranes with well-defined pore architectures. Here, we report a new approach to designing membranes with narrow molecular-sized channels and hydrophilic functionality that enable fast transport of salt ions and high size-exclusion selectivity towards small organic molecules. These membranes, based on polymers of intrinsic microporosity containing Tröger's base or amidoxime groups, demonstrate that exquisite control over subnanometre pore structure, the introduction of hydrophilic functional groups and thickness control all play important roles in achieving fast ion transport combined with high molecular selectivity. These membranes enable aqueous organic flow batteries with high energy efficiency and high capacity retention, suggesting their utility for a variety of energy-related devices and water purification processes.

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