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MOTIVATION: The annotation of cell types from single-cell transcriptomics is essential for understanding the biological identity and functionality of cellular populations. Although manual annotation remains the gold standard, the advent of automatic pipelines has become crucial for scalable, unbiased, and cost-effective annotations. Nonetheless, the effectiveness of these automatic methods, particularly those employing deep learning, significantly depends on the architecture of the classifier and the quality and diversity of the training datasets. RESULTS: To address these limitations, we present a Pruning-enabled Gene-Cell Net (PredGCN) incorporating a Coupled Gene-Cell Net (CGCN) to enable representation learning and information storage. PredGCN integrates a Gene Splicing Net (GSN) and a Cell Stratification Net (CSN), employing a pruning operation (PrO) to dynamically tackle the complexity of heterogeneous cell identification. Among them, GSN leverages multiple statistical and hypothesis-driven feature extraction methods to selectively assemble genes with specificity for scRNA-seq data while CSN unifies elements based on diverse region demarcation principles, exploiting the representations from GSN and precise identification from different regional homogeneity perspectives. Furthermore, we develop a multi-objective Pareto pruning operation (Pareto PrO) to expand the dynamic capabilities of CGCN, optimizing the sub-network structure for accurate cell type annotation. Multiple comparison experiments on real scRNA-seq datasets from various species have demonstrated that PredGCN surpasses existing state-of-the-art methods, including its scalability to cross-species datasets. Moreover, PredGCN can uncover unknown cell types and provide functional genomic analysis by quantifying the influence of genes on cell clusters, bringing new insights into cell type identification and characterizing scRNA-seq data from different perspectives. AVAILABILITY AND IMPLEMENTATION: The source code is available at https://github.com/IrisQi7/PredGCN and test data is available at https://figshare.com/articles/dataset/PredGCN/25251163.
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Análisis de la Célula Individual , Transcriptoma , Análisis de la Célula Individual/métodos , Transcriptoma/genética , Programas Informáticos , Anotación de Secuencia Molecular/métodos , Animales , Humanos , Perfilación de la Expresión Génica/métodos , Biología Computacional/métodos , AlgoritmosRESUMEN
BACKGROUND: The beneficial function of phytase and 25-hydroxyvitamin D3 (HyD) on the feed utilization rate has been widely investigated. However, studies concerning its influence on weaned piglets largely lag behind. The aim of this study was to investigate the effects of phytase and HyD supplementation on the growth performance and bone development in weaned piglets under dietary Ca and P deficiency. RESULTS: The results showed that dietary Ca and P deficiency decreased (P < 0.05) the content of serum P in 6-10 kg piglets, as well as reducing (P < 0.05) the contents of serum Ca and P, average daily gain (ADG), bone mineral density (BMD), breaking force (BF), bone ash and femur Ca in 10-20 kg piglets. Compared with the control group, the feed-to-gain ratio (F/G) of 6-10 kg piglets in the Phy group was decreased (P < 0.05), whereas the ADG, blood Ca and P, BMD, BF, bone ash, P apparent digestibility, Ca and P retention rate of 10-20 kg piglets were increased (P < 0.05). The contents of serum osteocalcin and HyD in 6-10 kg piglets and ADG were higher than in the control group (P < 0.05), as well as the contents of serum Ca and HyD in 10-20 kg piglets in the HyD treatment group. Supplementation with both Phy and HyD decreased the F/D (P < 0.05) and increased the contents of serum Ca, P and HyD in 6-10 kg piglets as well as enhancing the ADG, BMD, BF, bone ash, femur Ca and P, serum Ca and P, HyD, and the apparent digestibility and retention of Ca and P (P < 0.05) in 10-20 kg piglets. Supplementation with Phy and HyD in Ca- and P-deficient dietary decreased bone resorption, and improved tight arrangement of collagen fibers and oblique fibers in weaned piglets. CONCLUSION: These data indicated that supplementation with both 1500 U kg-1 Phy and 50 µg kg-1 HyD could enhance dietary Ca and P utilization and promote bone development in low Ca and P dietary, and supplementation with both Phy and HyD had a significant synergy effect compared to single supplement. © 2021 Society of Chemical Industry.
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6-Fitasa/metabolismo , Desarrollo Óseo , Calcifediol/metabolismo , Calcio/deficiencia , Fósforo/deficiencia , Porcinos/crecimiento & desarrollo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Densidad Ósea , Huesos/metabolismo , Dieta/veterinaria , Suplementos Dietéticos/análisis , Femenino , Masculino , Porcinos/metabolismoRESUMEN
Over 130 y have passed since Charles Darwin first discovered that the adventitious roots of English ivy (Hedera helix) exude a yellowish mucilage that promotes the capacity of this plant to climb vertical surfaces. Unfortunately, little progress has been made in elucidating the adhesion mechanisms underlying this high-strength adhesive. In the previous studies, spherical nanoparticles were observed in the viscous exudate. Here we show that these nanoparticles are predominantly composed of arabinogalactan proteins (AGPs), a superfamily of hydroxyproline-rich glycoproteins present in the extracellular spaces of plant cells. The spheroidal shape of the AGP-rich ivy nanoparticles results in a low viscosity of the ivy adhesive, and thus a favorable wetting behavior on the surface of substrates. Meanwhile, calcium-driven electrostatic interactions among carboxyl groups of the AGPs and the pectic acids give rise to the cross-linking of the exuded adhesive substances, favor subsequent curing (hardening) via formation of an adhesive film, and eventually promote the generation of mechanical interlocking between the adventitious roots of English ivy and the surface of substrates. Inspired by these molecular events, a reconstructed ivy-mimetic adhesive composite was developed by integrating purified AGP-rich ivy nanoparticles with pectic polysaccharides and calcium ions. Information gained from the subsequent tensile tests, in turn, substantiated the proposed adhesion mechanisms underlying the ivy-derived adhesive. Given that AGPs and pectic polysaccharides are also observed in bioadhesives exuded by other climbing plants, the adhesion mechanisms revealed by English ivy may forward the progress toward understanding the general principles underlying diverse botanic adhesives.
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Adhesivos/química , Hedera/química , Mucoproteínas/química , Secuencia de Aminoácidos , Secuencia de Bases , Calcio/química , Reactivos de Enlaces Cruzados , ADN de Plantas/genética , Hedera/genética , Microscopía de Fuerza Atómica , Modelos Moleculares , Estructura Molecular , Mucoproteínas/genética , Mucoproteínas/ultraestructura , Nanosferas/química , Nanosferas/ultraestructura , Pectinas/química , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/ultraestructura , HumectabilidadRESUMEN
While tremendous efforts have been made in investigating scalable approaches for fabricating nanoparticles, less progress has been made in scalable synthesis of cyclic peptide nanoparticles and nanotubes, despite their great potential for broader biomedical applications. In this paper, tunable synthesis of self-assembled cyclic peptide nanotubes and nanoparticles using three different methods, phase equilibrium, pH-driven, and pH-sensitive methods, were proposed and investigated. The goal is scalable nanomanufacturing of cyclic peptide nanoparticles and nanotubes with different sizes in large quality by controlling multiple process parameters. Cyclo-(L-Gln-D-Ala-L-Glu-D-Ala-)2 was applied to illustrate the proposed ideas. In the study, mass spectrometry and high performance liquid chromatography were employed to verify the chemical structures and purity of the cyclic peptides. Morphology and size of the synthesized nanomaterials were characterized using atomic force microscopy and dynamic light scattering. The dimensions of the self-assembled nanostructures were found to be strongly influenced by the cyclic peptide concentration, side chain modification, pH values, reaction time, stirring intensity, and sonication time. This paper proposed an overall strategy to integrate all the parameters to achieve optimal synthesis outputs. Mechanisms of the self-assembly of the cyclic peptide nanotubes and nanoparticles under variable conditions and tunable parameters were discussed. This study contributes to scalable nanomanufacturing of cyclic peptide based self-assembled nanoparticles and nanotubes for broader biomedical applications.
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Nanopartículas/química , Nanotecnología/métodos , Nanotubos de Péptidos/química , Secuencia de Aminoácidos , Técnicas de Química Sintética , Concentración de Iones de Hidrógeno , Cinética , SonicaciónRESUMEN
Oxidative stress has been associated with a number of physiological problems in swine, including reduced production efficiency. Recently, although there has been increased research into regulatory mechanisms and antioxidant strategies in relation to oxidative stress-induced pig production, it remains so far largely unsuccessful to develop accurate models and nutritional strategies for specific oxidative stress factors. Here, we discuss the dose and dose intensity of the causes of oxidative stress involving physiological, environmental and dietary factors, recent research models and the antioxidant strategies to provide theoretical guidance for future oxidative stress research in swine.
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Previous studies have shown that maternal resveratrol improved growth performance and altered the microbial composition of suckling piglets under hot summer conditions. However, it remains unclear how maternal resveratrol improves growth performance of suckling piglets during high summer temperatures. A total of 20 sows (Landrace × Large White; three parity) were randomly assigned to 2 groups (with or without 300 mg/kg resveratrol) from d 75 of gestation to d 21 of lactation during high ambient temperatures (from 27 to 30 °C). The results showed that maternal resveratrol supplementation increased total daily weight gain of piglets under hot summer conditions, which is consistent with previous studies. Furthermore, we found that maternal resveratrol improved the intestinal morphology and intestinal epithelial proliferation in suckling piglets. Dietary resveratrol supplementation affected the characteristics of exosome-derived microRNAs (miRNAs) in sow colostrum, as well as the genes targeted by differentially produced miRNAs. MiRNAs are concentrated in the tight junction pathway. As a result, the expression of intestinal tight junction proteins was increased in suckling piglets (P < 0.05). Notably, maternal resveratrol increased the intestinal secretory immunoglobulin A (sIgA) levels of suckling piglets via colostrum immunoglobin (P < 0.05), which could increase the abundance of beneficial microbiota to further increase the concentration of short chain fatty acids (SCFA) in suckling piglets' intestine (P < 0.05). Finally, our correlation analysis further demonstrated the positive associations between significantly differential intestinal microbiota, intestinal sIgA production and SCFA concentrations, as well as the positive relation between total daily weight gain and intestinal health of suckling piglets. Taken together, our findings suggested that maternal resveratrol could promote intestinal health to improve piglet growth during high summer temperatures, which might be associated with the immunoglobin and exosome-derived miRNAs in sows' colostrum.
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BACKGROUND: Naturally occurring nanoparticles isolated from English ivy (Hedera helix) have previously been proposed as an alternative to metallic nanoparticles as sunscreen fillers due to their effective UV extinction property, low toxicity and potential biodegradability. METHODS: This study focused on analyzing the physicochemical properties of the ivy nanoparticles, specifically, those parameters which are crucial for use as sunscreen fillers, such as pH, temperature, and UV irradiation. The visual transparency and cytotoxicity of ivy nanoparticles were also investigated comparing them with other metal oxide nanoparticles. RESULTS: Results from this study demonstrated that, after treatment at 100°C, there was a clear increase in the UV extinction spectra of the ivy nanoparticles caused by the partial decomposition. In addition, the UVA extinction spectra of the ivy nanoparticles gradually reduced slightly with the decrease of pH values in solvents. Prolonged UV irradiation indicated that the influence of UV light on the stability of the ivy nanoparticle was limited and time-independent. Compared to TiO2 and ZnO nanoparticles, ivy nanoparticles showed better visual transparency. Methylthiazol tetrazolium assay demonstrated that ivy nanoparticles exhibited lower cytotoxicity than the other two types of nanoparticles. Results also suggested that protein played an important role in modulating the three-dimensional structure of the ivy nanoparticles. CONCLUSIONS: Based on the results from this study it can be concluded that the ivy nanoparticles are able to maintain their UV protective capability at wide range of temperature and pH values, further demonstrating their potential as an alternative to replace currently available metal oxide nanoparticles in sunscreen applications.
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Cosméticos/química , Hedera/química , Nanopartículas de Magnetita/química , Protectores Solares/química , Fenómenos Químicos , Cosméticos/análisis , Calor , Concentración de Iones de Hidrógeno , Raíces de Plantas/química , Protectores Solares/análisis , Rayos Ultravioleta , Óxido de Zinc/análisis , Óxido de Zinc/químicaRESUMEN
Gold nanoparticles (AuNPs) have drawn significant interest in recent years due to unique properties that make them advantageous in biomedical applications, including drug delivery and tissue engineering. In this paper, we have developed multiple methods for the synthesis of AuNPs using English ivy as the substrate. In the first method, we have used actively growing English ivy shoots to develop a sustainable system for the production of ivy nanoparticles. The second method was developed using the extract from the adventitious roots of English ivy. The nanoparticles formed using both methods were compared to determine the size distribution, morphology, and chemical structure of the nanoparticles. Characterization of the AuNPs was conducted using ultraviolet-visible (UV-Vis) spectroscopy, dynamic light scattering (DLS), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS). In addition to the structural differences between the AuNPs formed from the different methods, details of the methods in terms of yield, duration, and speed of AuNP formation are also discussed. Further, this paper will show that AuNPs formed using both methods demonstrated efficient uptake in mammalian cells, which provides the potential for biomedical applications. The two methods developed through this research for eco-friendly synthesis of AuNPs present an alternative to traditional chemical synthesis methods.
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Oro/química , Hedera/metabolismo , Nanopartículas del Metal , Microscopía Electrónica de Rastreo , Espectrometría por Rayos X , Espectrofotometría UltravioletaRESUMEN
RNA-binding proteins play crucial roles in the regulation of gene expression, and understanding the interactions between RNAs and RBPs in distinct cellular conditions forms the basis for comprehending the underlying RNA function. However, current computational methods pose challenges to the cross-prediction of RNA-protein binding events across diverse cell lines and tissue contexts. Here, we develop HDRNet, an end-to-end deep learning-based framework to precisely predict dynamic RBP binding events under diverse cellular conditions. Our results demonstrate that HDRNet can accurately and efficiently identify binding sites, particularly for dynamic prediction, outperforming other state-of-the-art models on 261 linear RNA datasets from both eCLIP and CLIP-seq, supplemented with additional tissue data. Moreover, we conduct motif and interpretation analyses to provide fresh insights into the pathological mechanisms underlying RNA-RBP interactions from various perspectives. Our functional genomic analysis further explores the gene-human disease associations, uncovering previously uncharacterized observations for a broad range of genetic disorders.
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Proteínas de Unión al ARN , ARN , Humanos , ARN/genética , ARN/metabolismo , Proteínas de Unión al ARN/metabolismo , Sitios de Unión/genética , Unión Proteica , Secuenciación de Inmunoprecipitación de CromatinaRESUMEN
We propose and experimentally demonstrate a reflectance-based photonic crystal (PC) liquid sensor. The PC is made of two-dimensional TiO2 nanopillar arrays. Such a reflectance-based structure with large functional area not only simplifies the optical guiding but also enhances the sensor signal. A linear shift of reflectance peaks is found for liquids with refractive indices varying from 1.333 to 1.390 at wavelength near 1.5 µm. Excellent agreement between measured values and the generated reflectance model at a fixed wavelength is obtained, indicating the high potential of these PC-based liquid sensors for biological and environmental applications.
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Nanoestructuras , Fenómenos Ópticos , Fotones , Titanio , Carbohidratos/análisisRESUMEN
The cell-specific peptide TPS (TPSLEQRTVYAK) has been proposed as a potential candidate for fabricating tissue engineering scaffolds based on its ability of binding to human endothelial progenitor cells (EPC) with high affinity and specificity. In this study, the class I hydrophobin hgfI gene from Grifola frondosa and the tps were fused and cloned into pPIC9. The fusion gene was expressed in Pichia pastoris under the control of alcohol oxidase 1 promoter. Tricine-SDS-PAGE and Western blotting confirmed that the fusion protein TPS-linker-HGFI (TLH) was successfully secreted into the culture medium. The fusion protein TLH was purified by ultrafiltration and reverse-phase high performance liquid chromatography (RP-HPLC). Water contact angle (WCA) demonstrated that similar to recombinant HGFI (rHGFI), the purified TLH could convert the surface wettability of polystyrene and mica. X-ray photoelectron spectroscopy (XPS) measurements indicated that the purified TLH could form stable films on the hydrophobic siliconized glass surface. The cell adhesion examination showed that the TLH modified poly(ε-caprolactone) (PCL) could specially facilitate the EPC (particularly EPC derived from human) binding, while rHGFI modified PCL could nonselectively enhance cells adhesion. To the best of our knowledge, this is the first report that demonstrates that the TPS peptide was immobilized on biomaterial-PCL surface by fusion with hydrophobin. The potential application of this finding in combination with biomedical devices for EPC culture, will facilitate the current techniques used for cell-based therapies.
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Proteínas Fúngicas/biosíntesis , Grifola/genética , Oligopéptidos/metabolismo , Pichia/genética , Proteínas Recombinantes de Fusión/biosíntesis , Absorción , Animales , Adhesión Celular , Células Cultivadas , Clonación Molecular , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Grifola/metabolismo , Células Endoteliales de la Vena Umbilical Humana , Humanos , Oligopéptidos/química , Oligopéptidos/genética , Espectroscopía de Fotoelectrones , Pichia/metabolismo , Conejos , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Células Madre , HumectabilidadRESUMEN
A novel fabrication method for nanostructures made of TiO(2), a hard-to-etch material with very attractive optical, physical and chemical properties, is developed. This technique 'atomic layer deposition array defined by etch-back' (AARDE) enables the formation of a large area of perfectly ordered, high aspect ratio nanostructures, such as nanopillars, nanotubes and nanochannels. High quality functional surfaces and versatile structures with tunable dimensions on various substrates can be realized. With all the process steps being controllable and compatible with integrated circuits, high throughput and repeatability are achieved. To demonstrate the potential of this new technique, results for AARDE TiO(2) nanopillar arrays as photonic crystals are also reported.
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Background: Resveratrol has numerous beneficial properties, including antioxidant, anti-inflammatory, and immunomodulatory properties. High summer temperatures in Southern China affect the reproductive performance of sows. The present study aimed to investigate the effects of dietary resveratrol supplementation in different thermal environments on the reproductive performance, antioxidant capacity, immune function, and intestinal microbes of sows and piglets during late gestation and lactation, as well as their relationship with colostrum immunoglobulin. Methods: A two-phase experiment was conducted with 40 healthy multiparous sows. In the first phase of the experiment, 20 sows were used in a moderate temperature environment, and in the second phase of the experiment, the remaining 20 sows were used in a high-temperature environment. In both phases, sows were fed either a control diet or a diet consists of control diet and 300 mg/kg resveratrol starting on day 75 of gestation. Plasma, milk, and fecal samples were collected to obtain the indices of antioxidant capacity, immune function, and intestinal microbes. Results: The results showed that resveratrol supplementation increased the number of live births by 13.24 and 26.79% in the first and second phases, respectively, compared with the control group. In the second phase, resveratrol supplementation increased litter weight at weaning and in the concentrations of growth hormone (GH), insulin (INS), progesterone (PROG), triglycerides, and uric acid (UA). The plasma superoxide dismutase (SOD) level on day 110 of gestation and day 14 of lactation, as well as glutathione peroxidase (GSH-Px) on day 14 of lactation in the first phase, showed an increasing trend (p = 0.0728, p = 0.0932, and p = 0.067, respectively) in the resveratrol group, compared with the control group. On day 14 of lactation, the plasma total antioxidant capability (T-AOC) level was higher in the second phase, while the plasma malondialdehyde (MDA) level was lower in both phases in the resveratrol group. Resveratrol supplementation increased the abundance of immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) in colostrum and the relative abundance of Lactobacillus and Alloprevotella but decreased the relative abundance of Escherichia-shigella in piglet feces in the second phase. In addition, Spearman's correlation analysis indicated that the weight gain of weaned piglets was positively (p < 0.05) associated with IgM content in colostrum and the abundance of Lactobacillus in the fecal microbiota of piglets in the second phase. Moreover, the abundance of Alloprevotella was positively correlated with the contents of IgA and IgG in colostrum, while the abundance of Lactobacillus was positively correlated with IgM content. Conclusion: These findings indicated that maternal resveratrol supplementation could enhance the growth performance, antioxidant capacity, and intestinal health of piglets in a high temperature environment, which might be associated with increased immunoglobin secretion from colostrum.
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Deoxynivalenol (DON) reduces growth performance and damage intestinal function, and resveratrol (RES) has positive effects on growth performance and intestinal function. The purpose of this study was to investigate the protective mechanism of RES in vitro and vivo challenged with DON. The results showed that dietary supplementation with DON significantly increase the mRNA expression levels of mitophagy- related genes, and protein level for PINK1, Parkin, Beclin-1, Lamp, Atg5, Map1lc, Bnip3, Fundc1, Bcl2l1 and SQSTMS1 (P < 0.05), while supplementation with both RES and DON decreased those indexes in the ileum. Besides DON significantly decreased protein level for Pyruvate Dehydrogenase, Cytochrome c, MFN1, OPA1, and PHB1 (P < 0.05), while supplementation with both RES and DON increased protein level for PHB1, SDHA, and VDAC in the ileum. Moreover, in vitro, we found that DON significantly decreased mitochondrial respiration (P < 0.05), while RES + DON increased the rate of spare respiratory capacity. Also, DON significantly decreased total NAD and ATP (P < 0.05), while RES + DON increased the total NAD and ATP. These results indicate that RES may ameliorates the intestinal damage challenged with deoxynivalenol through mitophagy in weaning piglets.
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A high-visibility integrated optofluidic Mach-Zehnder interferometer based on liquid-core antiresonant reflecting optical waveguides is reported. The device's geometry has been optimized to minimize the intensity imbalance between the two arms for highly unbalanced Mach-Zehnder configurations. This results in a very compact device with a total length of only 2.5 mm and with required liquid volume of about 0.16 nl. High visibility is demonstrated for two interferometers corresponding to different sensing lengths. The devices have been optically characterized, and the measured interference fringes in the transmitted spectra show good agreement with the theoretical ones.
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Hydrophobins are small secreted proteins produced by filamentous fungi. Being amphipathic and self-assembling, hydrophobins have drawn great attention since their discovery. The increase of production can reduce the cost and open up several new applications of hydrophobins. We successfully expressed recombinant Class I hydrophobin HGFI (rHGFI) by using pPIC9 vector with an alcohol oxidase 1 promoter in Pichia pastoris. Tricine-SDS-PAGE and Western blotting demonstrated that rHGFI, an 8 kDa protein, was secreted into the culture medium. The culture conditions of the transformant strain were optimized by controlling the methanol concentration and induction time. Ultrafiltration and reverse-phase high performance liquid chromatography were used to perform a large-scale purification of rHGFI. A stable production of rHGFI around 86 mg/L was achieved after the two-step purification. X-ray photoelectron spectroscopy and water contact angle measurements indicated that the functional rHGFI could self-assemble on hydrophobic siliconized glass and Teflon as well as on hydrophilic mica surfaces. A methylthiazol tetrazolium assay showed that rHGFI film could facilitate human aortic smooth muscle cell proliferation due to its cytocompatibility.
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Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Grifola/genética , Pichia/genética , Supervivencia Celular , Células Cultivadas , Proteínas Fúngicas/aislamiento & purificación , Proteínas Fúngicas/metabolismo , Expresión Génica , Vectores Genéticos/genética , Grifola/química , Grifola/metabolismo , Humanos , Interacciones Hidrofóbicas e Hidrofílicas , Músculo Liso Vascular/citología , Espectroscopía de Fotoelectrones , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , UltrafiltraciónRESUMEN
Hydrophobins are small fungal proteins that self-assemble spontaneously at hydrophilic-hydrophobic interfaces and change the polar nature of the surfaces to which they attach. A new hydrophobin gene hgfI was identified recently from the edible mushroom Grifola frondosa. In this paper, the cloning, expression, purification, and polyclonal antibody preparation of the HGFI were described. The hgfI gene was cloned into pET-28a expression plasmid at the EcoRI and NdeI restriction sites and then transformed into Escherichia coli BL21 strain. SDSPAGE analysis showed that recombinant HGFI protein was satisfactorily expressed by optimizing the concentration and induction time of IPTG. The expressed recombinant HGFI protein was purified by electroelution because its inclusion body was insoluble in traditional processing method. After a desalting procedure with Sephadex G-25, the recombinant HGFI protein was used to immunize adult rabbits following standard protocol. ELISA and western blot analysis indicated that the produced antiserum could detect both HGFI protein expressed in the prokaryotic (E. coli) and in the eukaryotic cells (G. frondosa). Furthermore, the antiserum was used to determine the localization of HGFI protein in G. frondosa cells using an immunofluorescence technique. The results demonstrated that HGFI protein was localized in the cell wall, especially at the budding position of hypha. The polyclonal antibody against HGFI will facilitate further production and functional study of HGFI protein.
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Anticuerpos Antifúngicos/inmunología , Proteínas Fúngicas/biosíntesis , Proteínas Fúngicas/inmunología , Grifola/química , Proteínas Recombinantes de Fusión/biosíntesis , Proteínas Recombinantes de Fusión/inmunología , Animales , Anticuerpos Antifúngicos/aislamiento & purificación , Western Blotting , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Escherichia coli/genética , Proteínas Fúngicas/genética , Plásmidos/química , Conejos , Proteínas Recombinantes de Fusión/aislamiento & purificaciónRESUMEN
The microstructures, mechanical properties, and thermal conductivity (TC) of Al-2Fe-xCo (x = 0~0.8) alloys in as-cast, homogeneous annealed, and cool rolled states are systematically studied. Results indicate that appropriate Co modification (x ≤ 0.5) simultaneously improves the thermal and mechanical properties of as-cast Al-2Fe alloys. The improvement of TC is attributed to ameliorating the morphology of primary Al3Fe phases from needles to short rods and fine particles, which decreases the scattering probability of free electrons during the electronic transmission. However, further increasing the Co content (x = 0.8) decreases the TC due to the formation of a coarse plate-like Al2FeCo phase. Besides, the thermal conductivity of annealed Al-2Fe-xCo alloys is higher than that of as-cast alloys because of the elimination of lattice defects and spheroidization of Al3Fe phases. After cool rolling with 80 % deformation, thermal conductivity of alloys slightly increases due to the breaking down of Al2FeCo phases. The rolled Al-2Fe-0.3Co alloy exhibits the highest thermal conductivity, which is about 225 W/(m·K), approximately 11 % higher than the as-cast Al-2Fe sample. The ultimate tensile strength (UTS) and elongation (EL) of as-cast Al-2Fe-0.5Co (UTS: 138 MPa; EL: 22.0 %) are increased by 35 % and 69 %, respectively, compared with those of unmodified alloy (UTS: 102 MPa; EL: 13.0 %).
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Peptide nanostructures are biodegradable and are suitable for many biomedical applications. However, to be useful imaging probes, the limited intrinsic optical properties of peptides must be overcome. Here we show the formation of tryptophan-phenylalanine dipeptide nanoparticles (DNPs) that can shift the peptide's intrinsic fluorescent signal from the ultraviolet to the visible range. The visible emission signal allows the DNPs to act as imaging and sensing probes. The peptide design is inspired by the red shift seen in the yellow fluorescent protein that results from π-π stacking and by the enhanced fluorescence intensity seen in the green fluorescent protein mutant, BFPms1, which results from the structure rigidification by Zn(II). We show that DNPs are photostable, biocompatible and have a narrow emission bandwidth and visible fluorescence properties. DNPs functionalized with the MUC1 aptamer and doxorubicin can target cancer cells and can be used to image and monitor drug release in real time.
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Dipéptidos/farmacocinética , Colorantes Fluorescentes/farmacocinética , Nanopartículas/química , Neoplasias/patología , Imagen Óptica/métodos , Animales , Antineoplásicos/química , Antineoplásicos/farmacocinética , Línea Celular Tumoral , Dipéptidos/química , Doxorrubicina/química , Doxorrubicina/farmacocinética , Portadores de Fármacos/química , Portadores de Fármacos/farmacocinética , Liberación de Fármacos , Colorantes Fluorescentes/química , Humanos , Ratones , Neoplasias/metabolismoRESUMEN
An infusion-dialysis based procedure has been developed as an approach to isolate organic nanoparticles from green tea. Tea nanoparticle (TNP) can effectively load doxorubicin (DOX) via electrostatic and hydrophobic interactions. We established an ABCB1 overexpressing tumor xenograft mouse model to investigate whether TNP can effectively deliver DOX into tumors and bypass the efflux function of the ABCB1 transporter, thereby increasing the intratumoral accumulation of DOX and potentiating the anticancer activity of DOX. MTT assays suggested that DOX-TNP showed higher cytotoxicity toward CCD-18Co, SW620 and SW620/Ad300 cells than DOX. Animal study revealed that DOX-TNP resulted in greater inhibitory effects on the growth of SW620 and SW620/Ad300 tumors than DOX. In pharmacokinetics study, DOX-TNP greatly increased the SW620 and SW620/Ad300 intratumoral concentrations of DOX. But DOX-TNP had no effect on the plasma concentrations of DOX. Furthermore, TNP is a safe nanocarrier with excellent biocompatibility and minimal toxicity. Ex vivo IHC analysis of SW620 and SW620/Ad300 tumor sections revealed evidence of prominent antitumor activity of DOX-TNP. In conclusion, our findings suggested that natural nanomaterials could be useful in combating multidrug resistance (MDR) in cancer cells and potentiating the anticancer activity of chemotherapeutic agents in cancer treatment.