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Peptidoglycan recognition proteins (PGRPs) are a family of pattern recognition receptors that play a critical role in the immune response of invertebrates and vertebrates. Herein, the short ApPGRP-D gene was cloned from the model lepidopteran Antheraea pernyi. Quantitative PCR (qPCR) confirmed that ApPGRP-D is an immune-related protein and that the expression of ApPGRP-D can be induced by microorganisms. ApPGRP-D is a broad-spectrum pattern recognition protein that activates the prophenoloxidase cascade activation system and promotes the agglutination of microbial cells. Likely due to its amidase activity, ApPGRP-D can inhibit the growth of E. coli and S. aureus. In addition, we demonstrated for the first time that zinc ions, as important metal coenzymes, could promote multiple functions of ApPGRP-D but not its amidase activity.
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Proteínas de Transporte , Imunidade Humoral , Proteínas de Insetos , Mariposas , Animais , Mariposas/imunologia , Mariposas/genética , Mariposas/metabolismo , Mariposas/microbiologia , Proteínas de Insetos/metabolismo , Proteínas de Insetos/genética , Proteínas de Transporte/metabolismo , Proteínas de Transporte/genética , Escherichia coli , Staphylococcus aureus , Sequência de Aminoácidos , Antibacterianos/farmacologia , Catecol Oxidase/metabolismo , Clonagem Molecular , Zinco/metabolismo , Precursores EnzimáticosRESUMO
Groundwater environments are complex, and traditional advanced oxidation technologies mainly based on free radicals have limitations such as poor selectivity and low interference resistance, making it difficult to efficiently degrade target pollutants in groundwater. Therefore, we developed a sludge-based biochar-supported FeMg-layered double hydroxide catalyst (BC@FeMg-LDH) for the catalytic degradation of 2, 4-dichlorophenol (2, 4-DCP) using persulfate (PDS) as an oxidant. The removal efficiency of the catalyst exceeded 95%, showing high oxidation activity in a wide pH range while being almost unaffected by reducing substances and ions in the environment. Meanwhile, under neutral conditions, the leaching of metal ions from BC@FeMg-LDH was minimal, thereby eliminating the risk of secondary pollution. According to quenching experiments and electron paramagnetic resonance spectroscopy, the main active species during BC@FeMg-LDH/PDS degradation of 2, 4-DCP is 1O2, indicating a non-radical reaction mechanism dominated by 1O2. Characterization techniques, including X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy, revealed that the carbonyl (C = O) and metal hydroxyl (M-OH) groups on the material surface were the main reactive sites mediating 1O2 generation. The 1O2 generation mechanism during the reaction involved ketone-like activation of carbonyl groups on the biochar surface and complexation of hydroxyl groups on the material surface with PDS, resulting in the formation of O2·- and further generation of 1O2. 1O2 exhibited high selectivity toward electron-rich organic compounds such as 2, 4-DCP and demonstrated strong interference resistance in complex groundwater environments. Therefore, BC@FeMg-LDH holds promising applications for the remediation of organic-contaminated groundwater.
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Água Subterrânea , Hidróxidos , Hidróxidos/química , Carvão Vegetal/química , Metais , FenóisRESUMO
Although great success has been achieved in various computer vision tasks, deep neural networks (DNNs) suffer dramatic performance degradation when evaluated on out-of-distribution data. Domain generalization (DG) is proposed to handle this problem by learning domain-agnostic information from multiple source domains to generalize well on unseen target domains. Several methods resort to Fourier transform due to its simplicity and efficiency. They argue that amplitude spectra imply domain-specific information, which should be suppressed, while phase counterparts imply domain-agnostic information, which should be preserved. However, these methods only suppress the domain-specific information in source domains and neglect the relationship with target domains, leading to the persistence of the domain gap. Besides, these methods preserve domain-agnostic information by keeping phase components unchanged, causing them to be underutilized. In this paper, we propose Dual Branch Augmentation Module (DBAM) by leveraging Fourier transform and taking advantage of both amplitude and phase spectra. For the amplitude branch, we propose Inner-domain Amplitude Distribution Rectification (IADR) and Cross-domain Amplitude Dirichlet Mixup (CADM) to stabilize the training process and explore more feature space. In addition, we propose Test-time Amplitude Prototype Calibration (TAPC) to construct the connection between source and target domains during evaluation to further mitigate the domain gap. For the phase branch, we propose Random Symmetric Phase Perturbation (RSPP) to enhance the robustness for recognizing domain-agnostic information. With the combined contributions of the two branches, DBAM significantly surpasses other state-of-the-art (SOTA) methods. Extensive experiments on four benchmarks and further analysis demonstrate the effectiveness of DBAM.
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Generalização Psicológica , Aprendizagem , Benchmarking , Calibragem , Redes Neurais de ComputaçãoRESUMO
Rechargeable magnesium batteries (RMBs) are considered as one of the most promising candidates for next-generation batteries. However, the popularization of RMBs is seriously plagued due to the lack of suitable non-nucleophilic electrolytes and the passivation of Mg anode. Herein, a novel non-nucleophilic electrolyte is developed by introducing (s)-1-methoxy-2-propylamine (M4) into themagnesium aluminum chloride complex (MACC)-like electrolyte. The as-synthesizes Mg(AlCl4 )2 -IL-DME-M4 electrolyte enables robust reversible cycling of Mg plating/stripping with low overpotential, high anodic stability, and ionic conductivity (8.56 mS cm-1 ). These features should be mainly attributed to the in situ formation of an MgF2 containing Mg2+ -conducting interphase, which dramatically suppresses the passivation and parasitic reaction of Mg anode with electrolyte. Remarkably, the Mg/S batteries assemble with as-synthesize electrolyte and a new type MoS2 @CMK/S cathode deliver unprecedented electrochemical performance. Specifically, the Mg/S battery exhibited the highest reversible capacity up to 1210 mAh g-1 at 0.1 C, excellent rate capability and satisfactory long-term cycling stability with a reversible capacity of 370 mAh g-1 (coulombic efficiency of ≈100%) at 1.0 C for 600 cycles. The study findings provide a novel strategy and inspiration for designing efficient non-nucleophilic Mg electrolyte and suitable sulfur-host materials for practical Mg/S battery applications.
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Domain generalization (DG) aims to generalize from a large amount of source data that are fully annotated. However, it is laborious to collect labels for all source data in practice. Some research gets inspiration from semi-supervised learning (SSL) and develops a new task called semi-supervised domain generalization (SSDG). Unlabeled source data is trained jointly with labeled one to significantly improve the performance. Nevertheless, different research adopts different settings, leading to unfair comparisons. Moreover, the initial annotation of unlabeled source data is random, causing unstable and unreliable training. To this end, we first specify the training paradigm, and then leverage active learning (AL) to handle the issues. We further develop a new task called Active Semi-supervised Domain Generalization (ASSDG), which consists of two parts, i.e., SSDG and AL. We delve deep into the commonalities of SSL and AL and propose a unified framework called Gradient-Similarity-based Sample Filtering and Sorting (GSSFS) to iteratively train the SSDG and AL parts. Gradient similarity is utilized to select reliable and informative unlabeled source samples for these two parts respectively. Our methods are simple yet efficient, and extensive experiments demonstrate that our methods can achieve the best results on the DG datasets in the low-data regime without bells and whistles.
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Generalização Psicológica , Aprendizagem Baseada em Problemas , Transporte Proteico , Aprendizado de Máquina SupervisionadoRESUMO
Rapid corrosion and bacterial infection are obstacles to put into use biodegradable magnesium (Mg) alloy as biomedical materials. In this research, an amorphous calcium carbonate (ACC)@curcumin (Cur) loaded poly-methyltrimethoxysilane (PMTMS) coating prepared by self-assembly method on micro-arc oxidation (MAO) coated Mg alloy has been proposed. Scanning electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy and Fourier transform infrared spectroscopy are adopted to analyze the morphology and composition of the obtained coatings. The corrosion behaviour of the coatings is estimated by hydrogen evolution and electrochemical tests. The spread plate method without or with 808 nm near-infrared irradiation is applied to evaluate the antimicrobial and photothermal antimicrobial ability of the coatings. Cytotoxicity of the samples is tested by 3-(4,5)-dimethylthiahiazo(-z-y1)-2,5-di- phenytetrazoliumromide (MTT) and live/dead assay culturing with MC3T3-E1 cells. Results show that the MAO/ACC@Cur-PMTMS coating exhibited favourable corrosion resistance, dual antibacterial ability, and good biocompatibility. Cur was employed as an antibacterial agent and photosensitizer for photothermal therapy. The core of ACC significantly improved the loading of Cur and the deposition of hydroxyapatite corrosion products during degradation, which greatly promoted the long-term corrosion resistance and antibacterial activity of Mg alloys as biomedical materials.
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Curcumina , Corrosão , Antibacterianos , Ligas , Materiais Biocompatíveis , Magnésio , Carbonato de Cálcio , Materiais Revestidos BiocompatíveisRESUMO
Mn-based cathode material Li1.20Mn0.52Ni0.20Co0.08O2 was proposed and ameliorated by surface-coating poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) and doping Ga3+. X-ray diffraction and high-resolution transmission electron microscopy studies revealed that part of Ga3+ replacing the Ni site could reduce the Li+/Ni2+ mixing by forming a well-ordered layered structure and a homogeneous coating layer of PEDOT:PSS is covered on the surface of Li1.20Mn0.52Ni0.19Co0.08Ga0.01O2. The results of the electrochemical studies demonstrated the higher initial charging-discharging Coulombic efficiency, and outstanding rate capabilities and cyclic performance were obtained for the PEDOT:PSS-covered and Ga3+-doped samples. Especially, 2 wt % PEDOT:PSS-coated Li1.20Mn0.52Ni0.19Co0.08Ga0.01O2 delivered 38.3 mAh g-1, which is larger than the pristine cathode at a 5C high rate. Meanwhile, it could retain 189.6 mAh g-1 (90.3% of its initial discharge capacity at 45 °C) after 300 cycles with a 1C rate, while the pristine cathode only delivered 149.7 mAh g-1 with 80.7% cycling retention left. The results strongly suggested that such PEDOT:PSS-coated and Ga3+-doped Mn-based layered structure materials demonstrated high potential as a cathode candidate especially for high-energy applications.
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This work reports the design and preparation of novel organic (polyvinyl alcohol, PVA)-inorganic (neodymium nitrate, Nd(NO3)3) hybrid coatings on micro-arc oxidation (MAO) coating for magnesium (Mg) alloy corrosion protection. X-ray diffractometer, X-ray photoelectron spectroscopy, fourier transform infrared spectroscopy, field emission scanning electron microscope, Energy Dispersive X-ray spectrometer and surface roughness were applied to characterize the chemical composition and surface morphology of the coatings. The corrosion resistance of the coatings was evaluated by electrochemical and salt spray tests. The results suggested that the formation of PVA-Nd3+ and PVA-Mg2+ complexes promoted the enrichment of Nd3+ on the surface, and thereby improved the sealing quality and compactness of the coating. Interestingly, when the coating was damaged, the Nd3+ ions were transformed to their carbonates and covered the active sites, and thus exhibiting self-healing function. Further, the corrosion resistance of PVA-Nd3+ modified MAO composite coating on AZ31 Mg alloy was improved.
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Smart polymeric coatings with early corrosion self-warning and damage self-repairing characteristics have garnered tremendous interest due to their ability to sense corrosion reactions and repair coating defects. However, tracking the repair process and its underlying protection mechanism is highly challenging. Herein, we report the construction of a novel composite coating by incorporating multifunctional nanosensors (graphene oxide-zeolitic imidazole frameworks loaded with 1,10-phenanthroline) into a thermo-responsive polyurethane. Under damaging events, the localized acidity derived from metal corrosion stimulates the decomposition of the nanosensors to produce 1,10-phenanthroline and benzimidazole. The generated ferrous ions are rapidly sensed by the released 1,10-phenanthroline to produce a conspicuous red color, which warns of the corrosion occurrence. In profiting from the photothermal effect of graphene oxide, the composite coating exhibits efficient crack closure behavior under near-infrared light irradiation. Morphology observation indicates that a coating scratch (about 30 µm wide) almost closed with 20 s of irradiation. The photothermally activated crack closure combined with benzimidazole inhibition endows the prepared coating with superior self-repairing performance. Interestingly, the change in color intensity around the coating defect can assist in tracking the repair process. Therefore, this work provides a novel strategy to visualize microscopic behaviors during damage and repair processes.
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A novel l-histidine based ionic liquid (LHIL) was developed and successfully synthesized. Its structure was confirmed by Fourier-transform infrared spectroscopy, UV-vis spectroscopy, X-ray photoelectron spectroscopy, 1H-NMR and high-resolution mass spectrometry. The outstanding corrosion inhibition effect of the LHIL on mild steel in 1 M hydrochloric acid was thoroughly evaluated by Tafel plots, electrochemical impedance spectroscopy, and localized electrochemical strategies. The results revealed that the corrosion of mild steel was effectively suppressed by the adsorption of LHIL on its surface, and the best inhibition efficiency reached 98.8%. The adsorption behavior of LHIL on steel obeyed the Langmuir adsorption isotherm, which involved both chemisorption and physisorption. Theoretical calculations indicated the strong chemisorption of LHIL on steel, as proved by the low energy gap (ΔE = 0.0522 eV) and high binding energy (E binding = 303.47 kcal mol-1), which clearly confirmed the effectiveness of LHIL for steel protection.
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Surgical failures, caused by postoperative infections of bone implants, are commonly met, which cannot be treated precisely with intravenous antibiotics. Photothermal therapy (PTT) and photodynamic therapy (PDT) have attracted widespread attention due to their non-invasive antibacterial effects on tissues and no bacterial resistance, which may be an excellent approach to solve infections related to bone implants for biodegradable magnesium alloys. Herein, a sodium copper chlorophyllin (SCC) with a porphyrin ring induced Ca-P coating was prepared on AZ31 magnesium alloy via layer-by-layer (LbL) assembly. The morphology and composition of the samples were characterized through field emission scanning electron microscope (FE-SEM) with affiliated energy dispersive spectrometer (EDS), X-ray diffractometer (XRD), and Fourier infrared spectrometer (FTIR) and X-ray photoelectron spectrometer (XPS) as well. Potentiodynamic polarization, electrochemical impedance spectroscopy (EIS) and hydrogen evolution experiments were employed to evaluate the corrosion behavior of the samples. Atomic absorption spectrophotometer was used to measure Cu elemental content of different immersion periods. Cytocompatibility and antibacterial performance of the coatings were probed using in vitro cytotoxicity tests (MTT assay), live/dead cell staining and plate counting method. The results showed that the obtained (Ca-P/SCC)10 coating exhibited good corrosion resistance, antimicrobial activity (especially under 808 nm irradiation) and biocompatibility. The antibacterial rates for E. coli and S. aureus were 99.9% and 99.8%, respectively; and the photothermal conversion efficiency was as high as 42.1%. Triple antibacterial mechanisms including photodynamic, photothermal reactions and copper-ions release were proposed. This coating exhibited a promising application for biodegradable magnesium alloys.
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Polyaniline (PANI)/Epoxy copolymer as a core material was synthesized via a chemical oxidation method. Various analytical techniques, including scanning electron microscope, Fourier transform infrared spectroscopy, energy dispersive spectroscopy, thermogravimetry, and electrochemical impedance spectroscopy, were used to characterize the morphology, compositions, and self-healing and anticorrosion properties of the prepared microcapsules and coatings. The prepared PANI/Epoxy copolymer showed the best electrochemical corrosion resistance when the ratio of PANI to epoxy was 0.05: 1 (wt.:wt.). For the mass fraction of the core (PANI/Epoxy copolymer) of 60.84 ± 0.06 wt%, the mean particle diameter of the prepared microcapsules was 4.20 ± 0.92 µm. The coatings with 15 wt% microcapsules possessed excellent self-healing performance and corrosion resistance. The low-frequency impedance modulus at 0.01 Hz of scratched coatings immersed in the NaCl solution for 24 h was 5.27 × 106 Ω·cm2. Scratched self-repairing coating samples were able to resist corrosion for 384 h; thus, the microcapsules can be used to significantly extend the service life of the coatings. Microcapsule-containing PANI/Epoxy copolymers are expected to find use in anticorrosion coating systems, where the coatings could be applied directly on rusty steel surfaces.
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In unsupervised domain adaptation (UDA), many efforts are taken to pull the source domain and the target domain closer by adversarial training. Most methods focus on aligning distributions or features between the source domain and the target domain. However, little attention is paid to the interaction between finer-grained levels, such as classes or samples of the two domains. In contrast to UDA, another transfer learning task, i.e., few-shot learning (FSL), takes full advantage of the finer-grained-level alignment. Many FSL methods implement the interaction between samples of support sets and query sets, leading to significant improvements. We wonder whether we can get some inspiration from these methods and bring such ideas of FSL to UDA. To this end, we first take a closer look at the differences between FSL and UDA and bridge the gap between them by high-confidence sample selection (HCSS). Then we propose cross-attention map generation module (CAMGM) to interact samples selected by HCSS. Moreover, we propose a simple but efficient method called cross-attention-map-based regularization (CAMR) to regularize the feature maps generated by the feature extractor. Experiments on three challenging datasets demonstrate that CAMR can bring solid improvements when added to the original objective. More specifically, the proposed CAMR can outperform original methods by 1% to 2% in most tasks without bells and whistles.
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Atenção , AprendizagemRESUMO
Photoactivated sterilization has received more attention in dealing with implant-associated infections due to its advantages of rapid and effective bacteriostasis and broad-spectrum antibacterial activity. Herein, a micro-arc oxidation (MAO)/polymethyltrimethoxysilane (PMTMS)@hemin-induced calcium-bearing phosphate microsphere (Hemin-CaP) coating was prepared on pure magnesium (Mg) via MAO processing and dipping treatments. The morphology and composition of the coating were characterized via scanning electron microscopy, Fourier transform infrared spectrometer, X-ray diffractometer and X-ray photoelectron spectrometer. Corrosion behavior was evaluated through electrochemical and hydrogen evolution tests. The release of Fe3+ ions at different immersion times was measured with an atomic absorption spectrophotometer. Antibacterial performance and cytotoxicity were assessed using the spread plate method, MTT assay and live/dead staining experiment. The results showed that the corrosion current density of the MAO/PMTMS@(Hemin-CaP) coating (4.41 × 10-8 A·cm-2) was decreased by two orders of magnitude compared to that of pure Mg (3.12 × 10-6 A·cm-2). Photoactivated antibacterial efficiencies of the Hemin-CaP microspheres and MAO/PMTMS@(Hemin-CaP) coating reached about 99% and 92%, respectively, which we attributed to the photothermal and photodynamic properties of hemin with a porphyrin ring. Moreover, based on the release of Fe3+ ions, the MC3T3-E1 pre-osteoblasts' viability reached up to 125% after a 72 h culture, indicating a positive effect of the coating in promoting cell growth. Thus, this novel composite coating holds a promising application as bone implants.
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Embedding two-dimension micro/nanocontainers containing corrosion inhibitors into organic coating is a well-established concept to impart the coating with enhanced barrier and self-healing feature. Herein, a versatile nanoemulsion assembly approach was used to synthesis nanocarriers combing mesoporous polydopamine nanoparticles (MPDA) with reduced graphene oxide (GO), which was employed to encapsulate corrosion inhibitors (benzotriazole, BTA) to improve the anticorrosion performance of waterborne epoxy coating. The BTA release profiles from synthesized GO with MPDA (PDAG) demonstrated the rapid pH-triggered activities to acidic corrosion environment. With the addition of BTA-loaded PDAG, the composited epoxy coatings presented self-repairing behavior and enhanced corrosion resistance during long-term immersion. The outstanding anticorrosion performance is attributed to dual-protection mechanism provided by BTA-loaded PDAG: (1) MPDA endows GO with satisfactory interface compatibilities and thus provides impermeable barrier to delay the penetration process of corrosive electrolyte; (2) corrosion inhibitors including BTA and polydopamine form the adsorption layers on bare steel surface to resist continuous corrosion at metal/coating interface.
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Grafite , Nanopartículas , Indóis , PolímerosRESUMO
Metal substrates beneath polymeric coatings are susceptible to localized corrosion, which could result in lifetime reduction and catastrophic failure without timely repair treatment. In situ detection of corrosion and repair coating defects are in high demand yet challenging to fulfill so far. Herein, we report a smart polymeric coating by integrating nanosensors into the coating matrix, which is capable of efficient corrosion sensing and active anticorrosion protecting. The nanosensors were constructed by zeolitic imidazolate framework encapsulated with the polyethylene glycol-tannic acid complex. The morphology, chemical constitution, and stimulus responsiveness of nanosensors were systematically analyzed. The generation of local corrosion beneath coating can be promptly sensed and reported by a conspicuous purple color derived from tannic-iron ion coordinates. Meanwhile, local electrochemical impedance spectroscopy results proved that the metal degradation process at the defected interface can be largely inhibited, exhibiting active anticorrosion property. Furthermore, the constructed smart coating possessed superior impermeability and long-term protective performance under simulated seawater and harsh salts spray conditions. This feasible and effective strategy based on simple nanosensors to engineer smart coatings paves a new way to develop high environmental adaptability protective materials with protecting, corrosion sensing, and self-healing functions.
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The protective performance of graphene/polymer composite coatings largely depends on the interface design in resin matrix. Herein, we report the synthesis of bio-based cardanol epoxy modified graphene oxide (GODN) nanomaterial and its application in epoxy coatings for the achievement of fine interface toward high performance anticorrosion composite coatings. The chemical composition of prepared GODN nanomaterial was investigated by FTIR, Raman and XPS spectra, respectively. The presence of cardanol epoxy attached on GO surface promotes the formation of chemical bonds between GO and epoxy resin, providing strong interfacial interaction and enhanced adhesion. Electrochemical results revealed that the GODN1%/EP composite coating exhibits high impedance (4.38 × 108 Ω cm2) even after 45 days immersion. Compared with pure EP coating, the localized corrosion reaction of GODN1%/EP coating can be inhibited under defected interface. The enhanced protective performance of GODN/EP composite coating was attributed to two aspects: (1) the impermeable GO greatly suppressed the penetration of aggressive ions and (2) the attached cardanol epoxy chains effectively improved the interfacial interaction and thus inhibited the crack propagation.
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Grafite , Nanoestruturas , Corrosão , Resinas EpóxiRESUMO
Polymeric materials are susceptible to minor damage, which is undetectable. Without timely and effective repair treatment, the damage may deteriorate the integrity of the materials and ultimately result in material failure and catastrophe. Autonomous warning and simultaneous damage repair are of great practical significance yet difficult to realize. Herein, we introduce a smart coating with autonomous warning and repairing of damage by the simple incorporation of nanosensors embedded with phenanthroline as a corrosion indicator and inhibitor. The electrochemical corrosion resulting from coating damage can be rapidly indicated by a prominent orange-red color in just five minutes. In addition to the warning function, the smart coating exhibits efficient self-repairing in the defective region, as reflected from the disappearance of the electrochemical admittance peak. This simple and powerful strategy dependent on a single active component to achieve an autonomous warning and repairing effect is highly expected to provide a new avenue for enhancing the security and longevity of other polymeric materials.
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The high rigidity and brittleness of traditional thermosetting resin based on bisphenol epoxy limits its many potential technical applications. Here, a novel tertiary amine containing cardanol-based epoxy resin (NC-514-DEA) was synthesized by reaction of diethanolamine (DEA) with cardanol epoxy resin (NC-514). Moreover, NC-514-DEA modified graphene oxide (GOND) was prepared and used as a reactive nano-reinforcing filler for epoxy composites. The results show that, compared with neat epoxy resin, the fracture toughness of the epoxy composite with 0.5 wt% GOND is increased by nearly 10%, and the friction coefficient is reduced from 0.567 to 0.408, demonstrating the best performance among specimens. The improved mechanical and wear resistance properties of prepared composites were attribute to the synergistic effect of NC-514-DEA and GO, which inhibited the generation and propagation of cracks by enhancing the interfacial interaction and distributing stress. In addition, the synthetic process of GOND is green, simple and efficient, providing a novel way for designing epoxy composite materials with many potential applications.
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AIMS: Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is caused by germline mutations in the Fumarate hydratase (FH) gene. In young women, the syndrome often presents with symptomatic uterine leiomyomas, leading to myomectomy or hysterectomy. In this study, we aimed to investigate the incidence and mutational profiles of FH-negative leiomyomas from young patients, thus allowing for early identification and triage of syndromic patients for surveillance. METHODS AND RESULTS: We evaluated 153 cases of uterine leiomyomas from women aged up to 30 years for loss of FH expression by tissue microarray (TMA)-based immunohistochemical staining. Mutational analysis of tumours with loss of FH was carried out by polymerase chain reaction (PCR) amplification of 10 exons within the FH gene and subsequent Sanger sequencing. The status of promoter methylation was assessed by bisulphite sequencing. Loss of FH protein expression was detected in seven (4.6%) of 153 tested uterine leiomyomas from young patients. All FH-negative leiomyomas displayed staghorn vasculature and fibrillary/neurophil-like cytoplasm. We found that six (86%) of seven FH-negative tumours detected by immunohistochemistry harboured FH mutations, 50% of which contained germline mutations. In particular, the germline mutational rate in FH gene was 2.0% (three of 153 cases). Bisulphite sequencing analysis failed to detect promoter methylation in any of the seven tumours. CONCLUSION: Our study showed a relatively high rate of FH germline mutation in FH-negative uterine leiomyomas from patients aged up to 30 years. While genetic mutations confer protein expression loss, epigenetic regulation of the FH gene appears to be unrelated to this phenotype.