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PRPF19 seems to play either tumor-promoting or anti-tumor roles depending on cancer types. This study aimed to clarify the potential role and mechanism of PRPF19 in bladder cancer. PRPF19 expression and its correlation with patients' overall survival were analyzed in bladder cancer. The effects of PRPF19 on the viability, apoptosis, DNA damage repair, and gemcitabine sensitivity in human bladder cancer cells (T24 and 5637) were analyzed through loss- or gain-of-function methods. Moreover, the influences of DDB1 small interfering RNA on these indexes were evaluated in bladder cancer cells. At last, rescue experiment using DDB1 overexpression was carried out to confirm whether PRPF19 functioned via regulating DDB1. PRPF19 was highly expressed in bladder cancer tissues and cells. Elevated PRPF19 expression was related to shorter overall survival of bladder cancer patients. Downregulation of PRPF19 inhibited cell proliferation, promoted cell apoptosis, increased the number of γ-H2AX-positive cells, and reduced the mRNA and protein levels of DDB1 and BRCA1. Meanwhile, knockdown of PRPF19 decreased the IC50 of gemcitabine and promoted gemcitabine-induced cell apoptosis. Whereas, PRPF19 overexpression significantly decreased gemcitabine-induced apoptosis in bladder cancer cells. DDB1 downregulation suppressed cell proliferation and BRCA1 expression, but elevated the number of γ-H2AX-positive cells and gemcitabine sensitivity. Upregulation of DDB1 attenuated γ-H2AX-positive cell number, BRCA1 expression and IC50 of gemcitabine that were affected by PRPF19 silencing. In conclusion, PRPF19 expression was upregulated in bladder cancer. It promoted cell growth and DNA damage repair, and decreased gemcitabine sensitivity via positively regulating DDB1 expression. Supplementary Information: The online version contains supplementary material available at 10.1007/s10616-023-00599-7.
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In this work, we designed and fabricated Pd/Au bimetallic thin film electrodes with isolated Pd nanoparticles via underpotential deposition of copper on a gold substrate followed by in situ redox replace reaction in a Pd salt solution. The Pd/Au electrode was characterized by AFM and XPS as well as multiple electrochemical techniques including CV and electrochemical quartz crystal microbalance (EQCM) in sulfuric acid and phosphate buffer electrolytes. Results show that the reduction reactions of the analytes (i.e., H2O2 and 3-nitrotyrosine (3-NT)) at the Pd/Au thin film surfaces affect the nature and reactivity of Pd/Au surface electrochemistry including the adsorbed/absorbed hydrogen and/or the premonolayer palladium oxide redox processes at Pd. The EQCM experiment supports the arrangement of small size Pd nanoparticles in the Pd thin film in the presence of gold exhibits unusual properties, acting as a new physicochemical dimension between the electrode and target H2O2 and 3-NT molecules. The Pd/Au thin film was demonstrated as an extremely sensitive and selective probe for detection of common ROS and RNS (i.e., H2O2 and 3-NT). The integration of two different metallic species, Pd and Au, into a surface structure on nanoscale by exploiting their unique surface electrochemistry establishes an innovative analytical method for highly sensitive and selective detection of H2O2 and 3-NT simultaneously. This method has a general scope for detecting a broad range of redox active and nonredox active species simultaneously, which opens up new opportunities to develop new electrocatalytic materials and innovative sensing approaches.
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CD20, expressed in greater than 90% of B-lymphocytic lymphomas, is a target for antibody therapy. Rituximab is a chimeric therapeutic monoclonal antibody (mAb) against the protein CD20, allowing it to destroy B cells and to treat lymphoma, leukemia, transplant rejection, and autoimmune disorder. In this work, the binding of rituximab to self-assembled monolayers (SAMs) of peptide mimotopes of CD20 antigen was systematically characterized. Four peptide mimotopes of CD 20 antigen were selected from the literature and redesigned to allow their SAM immobilizations on gold electrodes through a peptide linker with cysteine. The bindings of these peptides with rituximab and control mAbs (trastuzumab and bevacizumab) were characterized by quartz crystal microbalance (QCM). Among the four peptide mimotopes initially selected, the peptide designated as CN-14 (CGSGSGSWPRWLEN) was the most selective and sensitive for rituximab binding. The CN-14 SAM was further characterized by ellipsometry and atomic force microscopy. The thickness of the CN-14 SAM film was approximately 32 Å, and the CN-14 SAM is suggested to be stabilized by a salt bridge of Arg-10 and Glu-13 between CN-14 peptides. The CN-14 salt bridge was evaluated by a series of modifications to the CN-14 peptide sequence and characterized by QCM. The CN-14 amide variant produced a better affinity to rituximab than CN-14 without a significant impact on selectivity. As the pKa of the Glu residue of CN-14 increased, the affinity of the SAM to rituximab increased, whereas the selectivity decreased. This was attributed to the weakening of the salt bridge between the CN-14 Arg-10 and Glu-13 at higher pKa values for Glu-13. Our study shows that peptide mimotopes have potential benefits in sensor applications, as the peptide-peptide interactions in the SAMs can be manipulated by the addition of functional groups to the peptide to influence the binding of target proteins.
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Antígenos CD20/química , Rituximab/química , Sequência de Aminoácidos , Antígenos CD20/metabolismo , Biomimética , Ouro/química , Modelos Biológicos , Dados de Sequência Molecular , Ligação Proteica , Rituximab/metabolismo , Propriedades de SuperfícieRESUMO
There has been much discussion on the need to develop fluorescent quantum dots (QDs) as ultracompact probes, with overall size profiles comparable to those of the genetically encoded fluorescent tags. In the use of conventional semiconductor QDs for such a purpose, the beautifully displayed dependence of fluorescence color on the particle diameter becomes a limitation. More recently, carbon dots have emerged as a new platform of QD-like fluorescent nanomaterials. The optical absorption and fluorescence emissions in carbon dots are not bandgap in origin, different from those in conventional semiconductor QDs. The absence of any theoretically defined fluorescence color-dot size relationships in carbon dots may actually be exploited as a unique advantage in the size reduction toward having carbon dots serve as ultracompact QD-like fluorescence probes. Here we report on carbon dots of less than 5 nm in the overall dot diameter with the use of 2,2'-(ethylenedioxy)bis(ethylamine) (EDA) molecules for the carbon particle surface passivation. The EDA-carbon dots were found to be brightly fluorescent, especially over the spectral range of green fluorescent protein. These aqueous soluble smaller carbon dots also enabled more quantitative characterizations, including the use of solution-phase NMR techniques, and the results suggested that the dot structures were relatively simple and better-defined. The potential for these smaller carbon dots to serve as fluorescence probes of overall sizes comparable to those of fluorescent proteins is discussed.
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Corantes Fluorescentes/química , Nanotecnologia/métodos , Nanotubos de Carbono/química , Pontos Quânticos/química , Animais , Carbono/química , Isótopos de Carbono/química , Fluorescência , Proteínas de Fluorescência Verde/química , Humanos , Espectroscopia de Ressonância Magnética , Teste de Materiais , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Nanopartículas/química , Tamanho da Partícula , Semicondutores , Espectrofotometria , Propriedades de SuperfícieRESUMO
Hexagonal boron nitride is a two-dimensional layered material that can be stable at 1,500 °C in air and will not react with most chemicals. Here we demonstrate large-scale, ultrathin, oxidation-resistant coatings of high-quality hexagonal boron nitride layers with controlled thicknesses from double layers to bulk. We show that such ultrathin hexagonal boron nitride films are impervious to oxygen diffusion even at high temperatures and can serve as high-performance oxidation-resistant coatings for nickel up to 1,100 °C in oxidizing atmospheres. Furthermore, graphene layers coated with a few hexagonal boron nitride layers are also protected at similarly high temperatures. These hexagonal boron nitride atomic layer coatings, which can be synthesized via scalable chemical vapour deposition method down to only two layers, could be the thinnest coating ever shown to withstand such extreme environments and find applications as chemically stable high-temperature coatings.
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Carbon dots (surface-passivated small carbon nanoparticles) are crosslinked to result in fluorescence probes containing one or multiple dots. For the single-dot probes, the crosslinking further stabilizes the dot structure, while for those packed with multiple dots, the individual probe imaging results demonstrate that the fluorescence properties are additive, with more dots for higher emission intensities in a proportional fashion, thus enabling the preparation of ultra-bright fluorescence probes.
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Carbono/química , Fluorescência , Corantes FluorescentesRESUMO
Graphene and hexagonal boron nitride (h-BN) have similar crystal structures with a lattice constant difference of only 2%. However, graphene is a zero-bandgap semiconductor with remarkably high carrier mobility at room temperature, whereas an atomically thin layer of h-BN is a dielectric with a wide bandgap of â¼5.9 eV. Accordingly, if precise two-dimensional domains of graphene and h-BN can be seamlessly stitched together, hybrid atomic layers with interesting electronic applications could be created. Here, we show that planar graphene/h-BN heterostructures can be formed by growing graphene in lithographically patterned h-BN atomic layers. Our approach can create periodic arrangements of domains with size ranging from tens of nanometres to millimetres. The resulting graphene/h-BN atomic layers can be peeled off the growth substrate and transferred to various platforms including flexible substrates. We also show that the technique can be used to fabricate two-dimensional devices, such as a split closed-loop resonator that works as a bandpass filter.
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Molecules of n-alkanethiols with methyl head groups typically form well-ordered monolayers during solution self-assembly for a wide range of experimental conditions. However, we have consistently observed that, for either carboxylic acid or thiol-terminated n-alkanethiols, under certain conditions nanografted patterns are generated with a thickness corresponding precisely to a double layer. To investigate the role of head groups for solution self-assembly, designed patterns of omega-functionalized n-alkanethiols were nanografted with systematic changes in concentration. Nanografting is an in situ approach for writing patterns of thiolated molecules on gold surfaces by scanning with an AFM tip under high force, accomplished in dilute solutions of desired ink molecules. As the tip is scanned across the surface of a self-assembled monolayer under force, the matrix molecules are displaced from the surface and are immediately replaced with fresh molecules from solution to generate nanopatterns. In this report, side-by-side comparison of nanografted patterns is achieved for different matrix molecules using AFM images. The chain length and head groups (i.e., carboxyl, hydroxyl, methyl, thiol) were varied for the nanopatterns and matrix monolayers. Interactions such as head-to-head dimerization affect the vertical self-assembly of omega-functionalized n-alkanethiol molecules within nanografted patterns. At certain threshold concentrations, double layers were observed to form when nanografting with head groups of carboxylic acid and dithiols, whereas single layers were generated exclusively for nanografted patterns with methyl and hydroxyl groups, regardless of changes in concentration.
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Alcanos/química , Microscopia de Força Atômica , Nanotecnologia/métodos , Compostos de Sulfidrila/química , Ácidos Carboxílicos/química , Ouro/química , Hidróxidos/química , Ácidos Palmíticos/química , Padrões de Referência , Solventes/química , Propriedades de Superfície , Água/químicaRESUMO
Particle lithography is a practical approach to generate millions of organosilane nanostructures on various surfaces, without the need for vacuum environments or expensive instrumentation. This report describes a stepwise chemistry route to prepare organosilane nanostructures and then apply the patterns as a spatially selective foundation to attach gold nanoparticles. Sites with thiol terminal groups were sufficiently small to localize the attachment of clusters of 2-5 nanoparticles. Basic steps such as centrifuging, drying, heating, and rinsing were used to generate arrays of regular nanopatterns. Close-packed films of monodisperse latex spheres can be used as an evaporative mask to spatially direct the placement of nanoscopic amounts of water on surfaces. Vapor phase organosilanes deposit selectively at areas of the surface containing water residues to generate nanostructures with regular thickness, geometry, and periodicity as revealed in atomic force microscopy images. The area of contact underneath the mesospheres is effectively masked for later synthetic steps, providing exquisite control of surface coverage and local chemistry. By judicious selection in designing the terminal groups of organosilanes, surface sites can be engineered at the nanoscale for building more complex structures. The density of the nanopatterns and surface coverage scale predictably with the diameter of the mesoparticle masks. The examples presented definitively illustrate the capabilities of using the chemistry of molecularly thin films of organosilanes to spatially define the selectivity of surfaces at very small size scales.
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The solution self-assembly of alpha,omega-alkanedithiols onto Au(111) was investigated using atomic force microscopy (AFM). A heterogeneous surface morphology is apparent for 1,8-octanedithiol and for 1,9-nonanedithiol self-assembled monolayers (SAMs) prepared by solution immersion as compared to methyl-terminated n-alkanethiols. Local views from AFM images reveal a layer of mixed molecular orientations for alpha,omega-alkanedithiols, which evidence surface structures with heights corresponding to both lying-down and standing-up orientations. For dithiol SAMs prepared by solution self-assembly, the majority of alpha,omega-alkanedithiol molecules chemisorb with both thiol end groups bound to the Au(111) surface with the backbone of the alkane chain aligned parallel to the surface. However, AFM images disclose that there are also islands of standing molecules scattered throughout the surface. To measure the thickness of alpha,omega-alkanedithiol SAMs with angstrom sensitivity, methyl-terminated n-alkanethiols with known dimensions were used as molecular rulers. Under conditions of spatially constrained self-assembly, nanopatterns of alpha,omega-alkanedithiols written by nanografting formed monolayers with heights corresponding to an upright configuration.
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Microscopia de Força Atômica/métodos , Nanotecnologia/métodos , Compostos de Sulfidrila/química , Adsorção , Físico-Química/métodos , Processamento de Imagem Assistida por Computador , Teste de Materiais , Nanopartículas Metálicas/química , Nanopartículas/química , Soluções/química , Propriedades de SuperfícieRESUMO
Using a scanning probe lithography method known as nanografting in conjunction with knowledge of self-assembly chemistry, regulation of the heterogeneity of self-assembled monolayers (SAMs) is demonstrated. While nanografting in single-component thiols produces areas of SAMs with designed geometry and size, nanofabrication in mixed thiol solution yields segregated domains. The reaction mechanism in nanografting differs significantly from self-assembly in mix-and-grow methods, as proven in systematic studies reported in this article and a companion paper of theoretical calculations of the nanografting process. Knowledge of the reaction pathways enables development of methods for shifting the interplay between the kinetics and thermodynamics in SAM formation, and thus the heterogeneity of mixed SAMs. By varying fabrication parameters, such as shaving speed, and reaction conditions, such as concentration and ratio of the components, the lateral heterogeneity can be adjusted ranging from near molecular mixing to segregated domains of several to tens of nanometers.