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SH-SY5Y is a human neuroblastoma cell line that can be differentiated into several neuronal phenotypes, depending on culture conditions. For this reason, this cell line has been widely used as an in vitro model of neurodegenerative conditions, such as Parkinson's disease (PD). However, most studies published to date used fetal bovine serum (FBS) as culture medium supplement for SH-SY5Y cell differentiation. We report on the testing of human platelet lysate (hPL) as a culture medium supplement to support SH-SY5Y cell culture. Both standard hPL and a fibrinogen-depleted hPL (FD-hPL) formulation, which does not require the addition of anticoagulants to culture media, promoted an increase in SH-SY5Y cell proliferation in comparison to FBS, without compromising metabolic activity. SH-SY5Y cells cultured in hPL or FD-hPL also displayed a higher number of neurite extensions and stained positive for MAP2 and synaptophysin, in the absence of differentiation stimuli; reducing hPL or FD-hPL concentration to 1% v/v did not affect cell proliferation or metabolic activity. Furthermore, following treatment with retinoic acid (RA) and further stimulation with brain-derived neurotrophic factor (BDNF) and nerve growth factor beta (NGF-ß), the percentage of SH-SY5Y cells stained positive for dopaminergic neuronal differentiation markers (tyrosine hydroxylase [TH] and Dopamine Transporter [DAT]) was higher in hPL or FD-hPL than in FBS, and gene expression of dopaminergic markers TH, DAT, and DR2 was also detected. Overall, the data herein presented supports the use of hPL to differentiate SH-SY5Y cells into a neuronal phenotype with dopaminergic features, and the adoption of FD-hPL as a fully xenogeneic free alternative to FBS to support the use of SH-SY5Y cells as a neurodegeneration model.
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Plaquetas , Técnicas de Cultura de Células , Diferenciação Celular , Proliferação de Células , Neurônios Dopaminérgicos , Neuroblastoma , Humanos , Proliferação de Células/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Neuroblastoma/metabolismo , Neuroblastoma/patologia , Linhagem Celular Tumoral , Plaquetas/metabolismo , Neurônios Dopaminérgicos/metabolismo , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/citologia , Técnicas de Cultura de Células/métodos , Meios de Cultura/química , Meios de Cultura/farmacologia , Tretinoína/farmacologia , FenótipoRESUMO
Tumor cells may develop alterations in glycosylation patterns during the initial phase of carcinogenesis. These alterations may be important therapeutic targets for lectins with antitumor action. This work aimed to evaluate the in vitro cytotoxicity of VML on tumor and non-tumor cells (concentration of 25 µg/mL and then microdiluted) and evaluate its in vivo toxicity at different concentrations (1.8, 3.5 and 7.0 µg/mL), using Drosophila melanogaster. Toxicity in D. melanogaster evaluated mortality rate, as well as oxidative stress markers (TBARS, iron levels, nitric oxide levels, protein and non-protein thiols). The cytotoxicity assay showed that VML had cytotoxic effect on leukemic lines HL-60 (IC50 = 3.5 µg/mL), KG1 (IC50 = 18.6 µg/mL) and K562 (102.0 µg/mL). In the toxicity assay, VML showed no reduction in survival at concentrations of 3.5 and 7.0 µg/mL and did not alter oxidative stress markers at any concentrations tested. Cytotoxicity of VML from HL-60, KG1 and K562 cells could arise from the interaction between the lectin and specific carbohydrates of tumor cells. In contrast, effective concentrations of VML against no-tumor cells human keratinocyte - HaCat and in the D. melanogaster model did not show toxicity, suggesting that VML is a promising molecule in vivo studies involving leukemic cells.
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Drosophila melanogaster , Lectinas , Animais , Humanos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Drosophila melanogaster/efeitos dos fármacos , Células HL-60 , Lectinas/farmacologia , Lectinas/toxicidade , Estresse Oxidativo/efeitos dos fármacosRESUMO
2D materials, given their form-factor, high surface-to-volume ratio, and chemical functionality have immense use in sensor design. Engineering 2D heterostructures can result in robust combinations of desirable properties but sensor design methodologies require careful considerations about material properties and orientation to maximize sensor response. This study introduces a sensor approach that combines the excellent electrical transport and transduction properties of graphite film with chemical reactivity derived from the edge sites of semiconducting molybdenum disulfide (MoS2) through a two-step chemical vapour deposition method. The resulting vertical heterostructure shows potential for high-performance hybrid chemiresistors for gas sensing. This architecture offers active sensing edge sites across the MoS2 flakes. We detail the growth of vertically oriented MoS2 over a nanoscale graphite film (NGF) cross-section, enhancing the adsorption of analytes such as NO2, NH3, and water vapor. Raman spectroscopy, density functional theory calculations and scanning probe methods elucidate the influence of chemical doping by distinguishing the role of MoS2 edge sites relative to the basal plane. High-resolution imaging techniques confirm the controlled growth of highly crystalline hybrid structures. The MoS2/NGF hybrid structure exhibits exceptional chemiresistive responses at both room and elevated temperatures compared to bare graphitic layers. Quantitative analysis reveals that the sensitivity of this hybrid sensor surpasses other 2D material hybrids, particularly in parts per billion concentrations.
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Understanding interactions between legacy and emerging environmental contaminants has important implications for risk assessment, especially when mutagens and carcinogens are involved, whose critical effects are chronic and therefore difficult to predict. The current work aimed to investigate potential interactions between benzo[a]pyrene (B[a]P), a carcinogenic polycyclic aromatic hydrocarbon and legacy pollutant, and diclofenac (DFC), a non-steroidal anti-inflammatory drug and pollutant of emerging concern, and how DFC affects B[a]P toxicity. Exposure to binary mixtures of these chemicals resulted in substantially reduced cytotoxicity in human HepG2 cells compared to single-chemical exposures. Significant antagonistic effects were observed in response to high concentrations of B[a]P in combination with DFC at IC50 and â IC50. While additive effects were found for levels of intracellular reactive oxygen species, antagonistic mixture effects were observed for genotoxicity. B[a]P induced DNA strand breaks, γH2AX activation, and micronuclei formation at ½ IC50 concentrations or lower, whereas DFC induced only low levels of DNA strand breaks. Their mixture caused significantly lower levels of genotoxicity by all three endpoints compared to those expected based on concentration additivity. In addition, antagonistic mixture effects on CYP1 enzyme activity suggested that the observed reduced genotoxicity of B[a]P was due to its reduced metabolic activation as a result of enzymatic inhibition by DFC. Overall, the findings further support the growing concern that co-exposure to environmental toxicants and their non-additive interactions may be a confounding factor that should not be neglected in environmental and human health risk assessment.
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Benzo(a)pireno , Carcinógenos Ambientais , Diclofenaco , Humanos , Diclofenaco/toxicidade , Benzo(a)pireno/toxicidade , Células Hep G2 , Carcinógenos Ambientais/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Ciclo-Oxigenase 1/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/farmacologia , Inibidores de Ciclo-Oxigenase/toxicidade , Ciclo-Oxigenase 2/metabolismo , Dano ao DNA/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase 2/farmacologia , Inibidores de Ciclo-Oxigenase 2/toxicidade , HistonasRESUMO
The current study investigates the venom-delivery system of green and red morphotypes of the sea anemone Actinia equina to disclose its potential as a source of bioactive compounds. We compared the two morphotypes using electron and optical microscopy, proteomics, and toxicity assessment on zebrafish embryos. Specialized venom-injecting cells (nematocysts) are equally distributed and found in the tentacles of both varieties. Proteomics revealed proteins of interest in both red and green Actinia, yielding the three most abundant Gene Ontology (GO) terms related to the biological processes "proteolysis", "hemolysis in another organism" and "lipid catabolic process". Neurotoxins and cytolytic toxins similar to known cnidarian toxins like PsTX-60A and AvTX-60A, for instance, were identified in both types. Extracts from green and red anemones were toxic to zebrafish embryos, with green anemone venom appearing to be more potent. The findings highlight the presence of proteinaceous toxins in A. equina and the potential for different varieties to possess distinct bioactive compounds. Notably, pore-forming toxins are suggested for molecular probes and immunotoxins, making them valuable assets for potential biotechnological and biomedical purposes.
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Proteinaceous toxins are peptides or proteins that hold great biotechnological value, evidenced by their ecological role, whether as defense or predation mechanisms. Bioprospecting using bioinformatics and omics may render screening for novel bioactives more expeditious, especially considering the immense diversity of toxin-secreting marine organisms. Eulalia sp. (Annelida: Phyllodocidae), a toxin bearing marine annelid, was recently shown to secrete cysteine-rich protein (Crisp) toxins (hitherto referred to as 'phyllotoxins') that can immobilize its prey. By analyzing and validating transcriptomic data, we narrowed the list of isolated full coding sequences of transcripts of the most abundant toxins or accompanying bioactives secreted by the species (the phyllotoxin Crisp, hyaluronidase, serine protease, and peptidases M12A, M13, and M12B). Through homology matching with human proteins, the biotechnological potential of the marine annelid's toxins and related proteins was tentatively associated with coagulative and anti-inflammatory responses for the peptidases PepM12A, SePr, PepM12B, and PepM13, and with the neurotoxic activity of Crisp, and finally, hyaluronidase was inferred to bear properties of an permeabilizing agent. The in silico analysis succeeded by validation by PCR and Sanger sequencing enabled us to retrieve cDNAs can may be used for the heterologous expression of these toxins.
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The properties of layered materials are significantly dependent on their lattice orientations. Thus, the growth of graphene nanowalls (GNWs) on Cu through PECVD has been increasingly studied, yet the underlying mechanisms remain unclear. In this study, we examined the GNWs/Cu interface and investigated the evolution of their microstructure using advanced Scanning transmission electron microscopy and Electron Energy Loss Spectroscopy (STEM-EELS). GNWs interface and initial root layers of comprise graphitic carbon with horizontal basal graphene (BG) planes that conform well to the catalyst surface. In the vertical section, the walls show a mix of graphitic and turbostratic carbon, while the latter becomes more noticeable close to the top edges of the GMWs film. Importantly, we identified growth process began with catalysis at Cu interface forming BG, followed by defect induction and bending at 'coalescence points' of neighboring BG, which act as nucleation sites for vertical growth. We reported that although classical thermal CVD mechanism initially dominates, growth of graphene later deviates a few nanometers from the interface to form GNWs. Nascent walls are no longer subjected to the catalytic action of Cu, and their development is dominated by the stitching of charged carbon species originating in the plasma with basal plane edges.
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Superconductivty has recently been induced in MXenes through surface modification. However, the previous reports have mostly been based on powders or cold-pressed pellets, with no known reports on the intrinsic superconsucting properties of MXenes at the nanoale. Here, it is developed a high-temperature atomic exchange process in NH3 atmosphere which induces superconductivity in either singleflakes or thin films of Nb2 CTx MXene. The exchange process between nitrogen atoms and fluorine, carbon, and oxygen atoms in the MXene lattice and related structural adjustments are studied using both experiments and density functional theory. Using either single-flake or thin-film devices, an anisotropic magnetic response of the 2D superconducting transformation has been successfully revealed. The anisotropic superconductivity is further demonstrated using superconducting thin films uniformly deposited over a 4 in. wafers, which opens up the possibility of scalable MXene-based superconducting devices.
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Graphene nanowalls (GNWs) can be described as extended nanosheets of graphitic carbon where the basal planes are perpendicular to a substrate. Generally, existing techniques to grow films of GNWsare based on plasma-enhanced chemical vapor deposition (PECVD) and the use of diverse substrate materials (Cu, Ni, C, etc) shaped as foils or filaments. Usually, patterned films rely on substrates priorly modified by costly cleanroom procedures. Hence, we report here the characterization, transfer and application of wafer-scale patterned GNWsfilms that were grown on Cu meshes using low-power direct-current PECVD. Reaching wall heights of â¼300 nm, mats of vertically-aligned carbon nanosheets covered square centimeter wire meshes substrates, replicating well the thread dimensions and the tens of micrometer-wide openings of the meshes. Contrastingly, the same growth conditions applied to Cu foils resulted in limited carbon deposition, mostly confined to the substrate edges. Based on the wet transfer procedure turbostratic and graphitic carbon domains co-exist in the GNWsmicrostructure. Interestingly, these nanoscaled patterned films were quite hydrophobic, being able to reverse the wetting behavior of SiO2surfaces. Finally, we show that the GNWscan also be used as the active material for C-on-Cu anodes of Li-ion battery systems.
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The immense biodiversity of marine invertebrates makes them high-value targets for the prospecting of novel bioactives. The present study investigated proteinaceous toxins secreted by the skin and proboscis of Glycera alba (Annelida: Polychaeta), whose congenerics G. tridactyla and G. dibranchiata are known to be venomous. Proteomics and bioinformatics enabled the detection of bioactive proteins that hold potential for biotechnological applications, including toxins like glycerotoxins (GLTx), which can interfere with neuromuscular calcium channels and therefore have value for the development of painkillers, for instance. We also identified proteins involved in the biosynthesis of toxins. Other proteins of interest include venom and toxin-related bioactives like cysteine-rich venom proteins, many of which are known to interfere with the nervous system. Ex vivo toxicity assays with mussel gills exposed to fractionated protein extracts from the skin and proboscis revealed that fractions potentially containing higher-molecular-mass venom proteins can exert negative effects on invertebrate prey. Histopathology, DNA damage and caspase-3 activity suggest significant cytotoxic effects that can be coadjuvated by permeabilizing enzymes such as venom metalloproteinases M12B. Altogether, these encouraging findings show that venomous annelids are important sources of novel bioactives, albeit illustrating the challenges of surveying organisms whose genomes and metabolisms are poorly understood.
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Anelídeos , Poliquetos , Toxinas Biológicas , Animais , Anelídeos/genética , Invertebrados , Organismos AquáticosRESUMO
Polychaeta are highly diversified invertebrates that inhabit marine, brackish or freshwater environments. They have acquired a unique range of adaptative features for securing food. However, the jaw apparatus may reveal not only defence and predation mechanisms, but also its relation to environmental chemistry. The present work compared the structure and chemical profile of the jaws of different estuarine Polychaeta: Nephtys hombergii (Nephtyidae), Hediste diversicolor (Nereididae) and Glycera alba (Glyceridae) using Scanning Electron Microscopy (SEM) and Scanning Electron Microscopy with Energy Dispersive X-Ray (SEM-EDX). Analyses revealed that N. hombergii possesses a muscular jawless proboscis with terminal sensorial papillae for detecting prey, whereas the G. alba proboscis exhibits four delicately sharp jaws with perforations for venom delivery and H. diversicolor bears two blunt denticulated jaws to grasp a wide variety of food items. Melanin and metals like copper provide hardness to the slender jaws of Glycera, while, in the absence of heavier metallic elements, halogens contribute to H. diversicolor jaws robustness. The more specific chemistry of the jaws of glycerids is associated with its more refined venom injection, whereas Hediste is an opportunistic omnivore and Nepthys an agile forager. Altogether, the chemistry of jaws is an adaptive feature for feeding, locomotion and even resilience to complex and often adverse chemical profiles of estuaries.
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Poliquetos , Animais , Microscopia Eletrônica de Varredura , Arcada Osseodentária , FaceRESUMO
In 2021, our research group published the prominent anticancer activity achieved through the successful combination of two redox centres (ortho-quinone/para-quinone or quinone/selenium-containing triazole) through a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The combination of two naphthoquinoidal substrates towards a synergetic product was indicated, but not fully explored. Herein, we report the synthesis of 15 new quinone-based derivatives prepared from click chemistry reactions and their subsequent evaluation against nine cancer cell lines and the murine fibroblast line L929. Our strategy was based on the modification of the A-ring of para-naphthoquinones and subsequent conjugation with different ortho-quinoidal moieties. As anticipated, our study identified several compounds with IC50 values below 0.5 µM in tumour cell lines. Some of the compounds described here also exhibited an excellent selectivity index and low cytotoxicity on L929, the control cell line. The antitumour evaluation of the compounds separately and in their conjugated form proved that the activity is strongly enhanced in the derivatives containing two redox centres. Thus, our study confirms the efficiency of using A-ring functionalized para-quinones coupled with ortho-quinones to obtain a diverse range of two redox centre compounds with potential applications against cancer cell lines. Here as well, it literally takes two for an efficient tango!
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Naftoquinonas , Quinonas , Animais , Camundongos , Quinonas/química , Benzoquinonas , Naftoquinonas/química , Oxirredução , Química Click , Reação de CicloadiçãoRESUMO
Cancer is one of the longest-known human diseases, yet only in recent times have we begun to perceive that the percentage of neoplasms caused by environmental factors, lifestyle and chemicals, is likely underestimated. The first medical reports associating cancer with pollutants like tars appeared by the early 20th century, but despite initial evidence relating oncogenesis and chromosomal alterations, only after the structure of DNA had been elucidated in the 1950s have genetic disorders been fully perceived as cause. This led to a growing interest in genotoxic and mutagenic pollutants. Even though we are now familiar with a range of environmental carcinogens spanning between aromatic hydrocarbons and asbestos to radionuclides and forms of carbon nanomaterials, establishing causal networks between pollutants and cancer remains cumbersome. In most part, this is due to the complexity of toxicant matrices, unknown modes-of-action of chemicals or their mixtures, the widening array of novel pollutants plus difficulties in subtracting background effects from true aetiology of disease. Recent advances in analytical chemistry, high-throughput toxicology, next-generation sequencing, computational biology and databases that allocate whole normal and cancer genomes, all indicate that we are on the verge of a new age of research into mechanistic 'oncotoxicology', but how can it impact risk assessment and prevention?
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Carcinógenos Ambientais , Poluentes Ambientais , Neoplasias , Humanos , Carcinógenos/toxicidade , Mutagênicos/toxicidade , Neoplasias/induzido quimicamente , Neoplasias/genética , Poluentes Ambientais/toxicidade , Carcinógenos Ambientais/toxicidade , CausalidadeRESUMO
The main goal was the development of a polysaccharide microcapsule for anticancer application based on guar gum and sodium alginate for the controlled release of hesperidin and betulinic acid by spray drying technique. The microcapsule showed an Encapsulation Efficiency of 98.15 ± 0.34 % for hesperidin and 99.76 ± 0.22 % for betulinic acid. In the release study, the Korsmeyer-Peppas mathematical model was identified as the most adequate to explain the observed release mechanism. In vivo tests were performed in zebrafish model, revealing that the microcapsules did not alter the locomotor activity and were not toxic within 96 h by oral administration, suggesting their biological safety. In vitro cytotoxic activity against HL-60 cells confirmed an IC50 value of 2.52 ± 0.23 µg mL-1 in 72 h. Additionally, a decrease in the cytotoxic activity of betulinic acid against L-929 (non-tumor) cells was evidenced. Therefore, the microcapsules synthesized in this work represent a promising formulation for anticancer applications.
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Alginatos , Hesperidina , Animais , Cápsulas , Peixe-Zebra , Ácido BetulínicoRESUMO
Mesenchymal stem cells (MSCs) hold promising therapeutic potential in several clinical applications, mainly due to their paracrine activity. The implementation of future secretome-based therapeutic strategies requires the use of easily accessible MSCs sources that provide high numbers of cells with homogenous characteristics. MSCs obtained from induced pluripotent stem cells (iMSCs) have been put forward as an advantageous alternative to the gold-standard tissue sources, such as bone marrow (BM-MSCs). In this study, we aimed at comparing the secretome of BM-MSCs and iMSCs over long-term culture. For that, we performed a broad characterization of both sources regarding their identity, proteomic secretome analysis, as well as replicative senescence and associated phenotypes, including its effects on MSCs secretome composition and immunomodulatory action. Our results evidence a rejuvenated phenotype of iMSCs, which is translated into a superior proliferative capacity before the induction of replicative senescence. Despite this significant difference between iMSCs and BM-MSCs proliferation, both untargeted and targeted proteomic analysis revealed a similar secretome composition for both sources in pre-senescent and senescent states. These results suggest that shifting from the use of BM-MSCs to a more advantageous source, like iMSCs, may yield similar therapeutic effects as identified over the past years for this gold-standard MSC source.
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Medula Óssea , Células-Tronco Mesenquimais , Diferenciação Celular , Proteômica , Secretoma , Senescência CelularRESUMO
Most animal toxins evolved to interact with specific molecular targets, which makes them highly-prized bioactives for drug development. Marine toxins, in particular, due to their wide chemical diversity, offer a new range of possibilities, a few of which have already been translated into approved drugs. Glycera alba and Hediste diversicolor are sympatric Polychaeta with distinct ecology and behavior suspected to secrete toxins that evolved to interact with distinct molecular targets, thus with differential selectivity and potential applications in drug discovery. Comparative transcriptomics revealed that while G. alba's venom apparatus is localized in the proboscis and neurotoxins are secreted to overtake prey, H. diversicolor secretes fewer and less specific toxins that are seemingly a defense. Human interactome-directed analysis unraveled novel toxins and other bioactives with potential biomedical applications, like proteins from G. alba's venom that can regulate apoptosis, whereas H. diversicolor yielded proteins that may control inflammation and cell proliferation in humans. Omics and bioinformatics appear to be powerful tools for marine bioprospecting and drug discovery, enabling molecular mining through transcriptomes of non-model organisms and link their ecology and physiology with protein's specificity and bioreactivity. Interactome-directed analysis against the human proteome seems an efficient alternative to the design of synthetic drugs.
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Poliquetos , Medicamentos Sintéticos , Animais , Descoberta de Drogas , Humanos , Toxinas Marinhas/metabolismo , Poliquetos/genética , Poliquetos/metabolismo , Proteoma/metabolismo , Medicamentos Sintéticos/metabolismo , TranscriptomaRESUMO
The vast ocean holds many unexplored organisms with unique adaptive features that enable them to thrive in their environment. The secretion of fluorescent proteins is one of them, with reports on the presence of such compounds in marine annelids being scarce. The intertidal Eulalia sp. is an example. The worm secretes copious amounts of mucus, that when purified and concentrated extracts, yield strong fluorescence under UV light. Emission has two main maxima, at 400 nm and at 500 nm, with the latter responsible for the blue-greenish fluorescence. Combining proteomics and transcriptomics techniques, we identified ubiquitin, peroxiredoxin, and 14-3-3 protein as key elements in the mucus. Fluorescence was found to be mainly modulated by redox status and pH, being consistently upheld in extracts prepared in Tris-HCl buffer with reducing agent at pH 7 and excited at 330 nm. One of the proteins associated with the fluorescent signal was localized in secretory cells in the pharynx. The results indicate that the secretion of fluorescent proteinaceous complexes can be an important defense against UV for this dweller. Additionally, the internalization of fluorescent complexes by ovarian cancer cells and modulation of fluorescence of redox status bears important considerations for biotechnological application of mucus components as markers.
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Anelídeos , Poliquetos , Animais , Biotecnologia , Corantes/metabolismo , Humanos , Muco/química , Extratos Vegetais/análise , Poliquetos/química , Proteínas/análiseRESUMO
Graphite sheets are known to exhibit remarkable performance in applications such as heating panels and critical elements of thermal management systems. Industrial-scale production of graphite films relies on high-temperature treatment of polymers or calendering of graphite flakes; however, these methods are limited to obtaining micrometer-scale thicknesses. Herein, we report the fabrication of a flexible and power-efficient cm2-scaled heater based on a polycrystalline nanoscale-thick graphite film (NGF, â¼100 nm thick) grown by chemical vapor deposition. The stability of these NGF heaters (operational in air over the range 30-300 °C) is demonstrated by a 12-day continuous heating test, at 215 °C. The NGF exhibits a fast switching response and attains a steady peak temperature of 300 °C at a driving bias of 7.8 V (power density of 1.1 W/cm2). This excellent heating performance is attributed to the structural characteristics of the NGF, which contains well-distributed wrinkles and micrometer-wide few-layer graphene domains (characterized using conductive imaging and finite element methods, respectively). The efficiency and flexibility of the NGF device are exemplified by externally heating a 2000 µm-thick Pyrex glass vial and bringing 5 mL of water to a temperature of 96 °C (at 2.4 W/cm2). Overall, the NGF could become an excellent active material for ultrathin, flexible, and sustainable heating panels that operate at low power.
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The present study investigated the potential toxicity of venomous secretions of two polychaetes, Hediste diversicolor and Glycera alba (Annelida: Phyllodocida). Toxic activity of putative toxins, measured on mussel gills through the Comet assay, revealed higher effects caused by extracts from H. diversicolor skin and G. alba specialised, jawed proboscis, when compared to control. The results suggest that H. diversicolor secretes toxins via skin for protection against predators, contrarily to G. alba, who secretes toxins for predation.
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In recent years, graphene has been explored as a heating membrane for studying high-temperature dynamics inside the transmission electron microscope (TEM) due to several limitations with the existing silicon nitride-based membrane. However, the transfer of monolayer graphene films for TEM experiments is challenging and requires many complicated steps with a minimum success rate. This work developed a novelin situheating platform by combining the graphene oxide (GO) flakes in the pre-patterned chips. The isolated GO flake was self-suspended between the metal electrodes by a simple drop-casting process. The GO was reduced and characterized using Raman and electron energy-loss spectroscopy. Furthermore, a GO-based heater was used to investigate the thermal stability of gold and silica nanoparticles. The gold nanoparticles evaporated non-uniformly and left an empty carbon shell, while silica disappeared uniformly by etching carbon support. We successfully demonstrated a GO flake as a heating membrane to study high temperature thermal dynamic reactions: melting/evaporation, agglomeration, Rayleigh instability, and formation/or removal of carbon in the nanoparticles.