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This work innovatively used cellulose nanofibers as a photocatalyst carrier, which could recycle nano-photocatalysts and minimize nanoparticle aggregation. The morphology, structures, chemical composition, optical-electronic properties and photocatalytic performance of amino-modified carbon quantum dots-ZnO/cellulose nanofiber (N-CQDs-ZnO/CNF: ZCH-2) hydrogel were characterized by SEM, TEM, BET, EDS, XRD, FTIR, UV-vis, XPS, PL and other techniques. The mechanism of Cr(VI) adsorption synergistic photoreduction by ZCH-2 was discussed in detail. The results showed that the prepared ZCH-2 had excellent removal performance for Cr(VI). After 120 min of adsorption and 40 min of photoreduction, the removal efficiency of Cr(VI) was 98.9 %. Compared with ZnO/CNF hydrogel, the adsorption performance of ZCH-2 increased by 268 % and the photoreduction performance increased by 116 %. The adsorption of Cr(VI) by ZCH-2 was controlled by electrostatic attraction and chemical adsorption. The photoreduction kinetic constant of ZCH-2 was 0.106 min-1, which was 8.9 times that of ZnO/CNF hydrogel. The N-CQDs in ZCH-2 could form N-CQDs-metal complexes with Cr(VI), resulting in fluorescence quenching, so Cr(VI) could be visually identified by fluorescence changes. This study provides a new idea for the design and optimization of a new multifunctional hydrogel with efficient adsorption-photoreduction-fluorescence recognition.
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Nanofibras , Pontos Quânticos , Poluentes Químicos da Água , Óxido de Zinco , Carbono/química , Óxido de Zinco/química , Nanofibras/química , Adsorção , Hidrogéis , Fluorescência , Poluentes Químicos da Água/química , Cromo/química , Celulose , CinéticaRESUMO
Intestinal mucus is the first line of defense against pathogens and has several active components. Poultry have a short intestine, the mucus of which may contain antiviral components. We hence investigated the antiviral components of mucus and explored their mechanisms of action. Initially, we isolated chicken intestinal mucus proteins that significantly inhibited the replication of avian viruses. The ileum 10-30 kDa protein fraction showed the greatest inhibition of viral replication. Moreover, liquid chromatography-mass spectrometry revealed 12 high-abundance proteins in the ileum 10-30 kDa protein fraction. Among them, we investigated the antiviral activity of calcium binding protein 1 (CALB1). Furthermore, eukaryotically and prokaryotically expressed CALB1 significantly suppressed the replication of avian viruses, possibly by binding calcium ions and/or inducing autophagy. In conclusion, we isolated and identified CALB1 from chicken intestinal mucus, which suppressed replication of avian viruses by regulating cellular calcium-ion homeostasis and autophagy.
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Cálcio , Galinhas , Animais , Íleo , Antivirais/farmacologia , MucoRESUMO
The gamma-coronavirus infectious bronchitis virus (IBV) has a high mutation rate and mainly invades the respiratory mucosa, making it difficult to prevent and causing great economic losses. Nonstructural protein 16 (NSP16) of IBV QX also not only plays an indispensable role in virus invading but also might hugely influence the antigen's recognition and presentation ability of host BMDCs. Hence, our study tries to illustrate the underline mechanism of how NSP16 influences the immune function of BMDCs. Initially, we found that NSP16 of the QX strain significantly inhibited the antigen presentation ability and immune response of mouse BMDCs, which was stimulated by Poly (I:C) or AIV RNA. Besides mouse BMDCs, we also found that NSP16 of the QX strain also significantly stimulated the chicken BMDCs to activate the interferon signaling pathway. Furthermore, we preliminarily demonstrated that IBV QX NSP16 inhibits the antiviral system by affecting the antigen-presenting function of BMDCs.
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Infecções por Coronavirus , Vírus da Bronquite Infecciosa , Doenças das Aves Domésticas , Animais , Camundongos , Galinhas , Apresentação de Antígeno , Infecções por Coronavirus/prevenção & controle , Infecções por Coronavirus/veterinária , Interferons , Doenças das Aves Domésticas/prevenção & controleRESUMO
Background: Osteoarthritis (OA) is a common chronic disease with numerous and interacting influencing factors, and current inadequate patient perceptions and behaviors in access to care contribute to the difficulties in the diagnosis, treatment, and management of osteoarthritis. Objective: The purpose of this study was to investigate the influencing factors of osteoarthritis (OA) in a southern Chinese population and to provide a scientific basis for the prevention and treatment of OA. Methods: A 1 : 2 matched case-control study was used to select 160 patients with OA from three hospitals in southern China as a case group. Three hundred and twenty cases of the same sex and similar age (within ± 2 years) were selected as the control group, and relevant data were collected for univariate and multivariate conditional logistic regression analysis. Results: There were no significant differences between the two groups of participants in terms of age, sex, and education (P > 0.05). Logistic regression statistical analysis showed that genetic factors (OR = 4.52, 95% CI = 1.56-7.83), body mass index (OR = 2.57, 95% CI = 1.16-5.84), alcohol consumption (OR = 3.81, 95% CI = 1.53-5.87), and a history of external joint limb injury (OR = 3.37, 95% CI = 1.67-5.24) would increase the risk of OA. In contrast, eating more fresh vegetables (OR = 0.08, 95% CI = 0.03-0.31), more fresh fruits (OR = 0.34, 95% CI = 0.12-0.96), more soy products (OR = 0.11, 95% CI = 0.04-0.45), and exposure to sunlight (OR = 0.31, 95% CI = 0.14-0.71) would reduce the OA risk of OA. Conclusion: Obesity, alcohol consumption, and a history of joint trauma all increase the risk of OA in a southern Chinese population, whereas a diet rich in fresh vegetables, fresh fruit, soy products, and sun exposure would reduce the risk of OA. In the future, we should focus on improving patients' awareness of medical care and developing their self-management skills, improving GPs' treatment skills, improving negative attitudes of both doctors and patients, and promoting positive patient care.
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Gallic acid is widely used in the field of food and medicine due to its diversified bioactivities. The extraction method with higher specificity and efficiency is the key to separate and purify gallic acid from complex biological matrix. Herein, using self-made core-shell magnetic molecularly imprinted polymers (MMIP) with gallic acid as template, a hollow magnetic molecularly imprinted polymer (HMMIP) with double imprinting/adsorption surfaces was prepared by etching the mesoporous silica intermediate layer of MMIP. The characterization and adsorption research showed that the HMMIP had larger specific surface area, higher magnetic response strength and a more stable structure, and the selectivity and saturated adsorption capacity (2.815 mmol/g at 318 K) of gallic acid on HMMIP were better than those of MMIP. Thus, in addition to MMIP, the improved HMMIP had excellent separation and purification ability to selectively extract gallic acid from complex matrix with higher specificity and efficiency.
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Selenocysteine (Sec) is recognized as the 21st amino acid employing as an essential building block for selenoproteins (SePs), which plays a significant role in various physiological processes. Therefore, there is an urgent need to reasonably develop some reliable and rapid methods for Sec detection in biological systems. In this work, we reported a new two-photon (TP) fluorescent probe BNT-Sec for Sec detection and imaging in living cells and zebrafish with two part: (1) a D-π-A-structured naphthalene derivative as a TP fluorophore; (2) a well-know Sec responsive site with strong intromolecular charge transfer effect (ICT) to selectively detect endogenous and exogenous. In the presence of Sec, probe BNT-Sec can initiate a Se-dependent specific aromatic nucleophilic substitution reaction, which exhibited BNT-Sec had a large fluorescence intensity enhancement with ~18.9-fold at 510 nm, a high sensitivity low LOD value' 10.6 nM, good light stability, strong specificity, pH stability and low cytotoxicity. In addition, BNT-Sec can be conveniently used to detect Sec in living cells and zebrafish for TP imaging due to the great TP measurement properties of fluorophore, exhibiting it has the potential to reveal the role of selenocysteine in physiological and pathological processes in further biological applications.
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Corantes Fluorescentes , Selenocisteína , Animais , Células HeLa , Humanos , Imagem Óptica , Fótons , Peixe-ZebraRESUMO
Osteosarcoma represents one of the most devastating cancers due to its high metastatic potency and fatality. Osteosarcoma is insensitive to traditional chemotherapy. Identification of a small molecule that blocks osteosarcoma progression has been a challenge in drug development. Phillygenin, a plant-derived tetrahydrofurofuran lignin, has shown to suppress cancer cell growth and inflammatory response. However, how phillygenin plays functional roles in osteosarcoma has remained unveiled. In this study, we showed that phillygenin inhibited osteosarcoma cell growth and motility in vitro. Further mechanistic studies indicated that phillygenin blocked STAT3 signaling pathway. Phillygenin led to significant downregulation of Janus kinase 2 and upregulation of Src homology region 2 domain-containing phosphatase 1. Gene products of STAT3 regulating cell survival and invasion were also inhibited by phillygenin. Therefore, our studies provided the first evidence that phillygenin repressed osteosarcoma progression by interfering STAT3 signaling pathway. Phillygenin is a potential candidate in osteosarcoma therapy.
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Antineoplásicos/farmacologia , Janus Quinase 2/metabolismo , Lignanas/farmacologia , Osteossarcoma/patologia , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Metástase NeoplásicaRESUMO
Hypochlorous acid (HClO) is a special kind of reactive oxygen species, which plays an important role in resisting pathogen invasion and maintaining cell redox balance and other physiological processes. In addition, HClO is commonly used in daily life as a bleaching and disinfectant agent. Its excessive use can also lead to death of water animals and serious respiratory and skin diseases in humans. Therefore, it is of great significance to develop a quick and convenient tool for detecting HClO in the environment and organisms. In this paper, we utilize the specific reaction of HClO with dimethylthiocarbamate to develop a novel naphthalene derivative fluorescent probe (BNA-HClO), it was designed and synthesized by using 6-(2-benzothiazolyl)-2-naphthol as the fluorophore and N,N-dimethylthiocarbamate as the recognition group. BNA-HClO shows large fluorescence enhancement (374-fold), high sensitivity (a detection limit of 37.56 nM), rapid response (<30 s), strong anti-interference ability and good specificity in vitro. Based on the outstanding in vitro sensing capability of BNA-HClO, it has been successfully used to detect spiked HClO in tap water, medical wastewater and fetal bovine serum with good recovery. BNA-HClO has also been successfully used as a portable test strip for the in situ semi-quantitative detection of HClO in tap water solutions. In addition, BNA-HClO can successfully enable the detection and imaging of exogenous and endogenous HClO in living cells. This work provides a simple and effective tool for the detection and imaging of HClO in environmental and biological systems, and provides some theoretical guidance for future exploration of biological and pathological studies related to HClO.
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Herein, we found that surface defects quench persistent luminescence in nanophosphors. Passivation of surface defects by thermal treatment or surface coating can effectively enhance the intensity and prolong the decay time of persistent luminescence. The surface passivated persistent nanophosphors are promising in autofluorescence-free bioimaging and time-gated steganography.
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The rapid detection of organophosphorus pesticide residues in food is crucial to food safety. One type of novel, magnetic, molecularly-imprinted polymeric microsphere (MMIP) was prepared with vinyl phosphate and 1-octadecene as a collection of dual functional monomers, which were screened by Gaussian09W molecular simulation. MMIPs were used to enrich organic phosphorus, which then detected by fluorescence quenching in vinyl phosphate-modified carbon dots (CDs@VPA) originated from anhydrous citric acid. MMIPs and CDs@VPA were characterized by TEM, particle size analysis, FT-IR, VSM, XPS, adsorption experiments, and fluorescence spectrophotometry in turn. Through the fitting data from experiment and Gaussian quantum chemical calculations, the molecular recognition properties and the mechanism of fluorescence detection between organophosphorus pesticides and CDs@VPA were also investigated. The results indicated that the MMIPs could specifically recognize and enrich triazophos with the saturated adsorption capacity 0.226 mmol g-1, the imprinting factor 4.59, and the limit of recognition as low as 0.0006 mmol L-1. Under optimal conditions, the CDs@VPA sensor has shown an extensive fluorescence property with a LOD of 0.0015 mmol L-1 and the linear range from 0.0035 mmol L-1 to 0.20 mmol L-1 (R2 = 0.9988) at 390 nm. The mechanism of fluorescence detection of organic phosphorus with CDs@VPA sensor might be attributable to hydrogen bonds formed between heteroatom O, N, S, or P, and the O-H group, which led to fluorescent quenching. Meanwhile, HN-C=O and Si-O groups in CDs@VPA system might contribute to cause excellent blue photoluminescence. The fluorescence sensor was thorough successfully employed to the detection of triazophos in cucumber samples, illustrating its tremendous value towards food sample analysis in complex matrix.
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The rapid detection of pollutants with high sensitivity and selectivity is of considerable significance for security screening, environmental safety, and human health. In this study, we prepared persistent luminescence nanoparticles (PLNPs) and constructed a label-free sensor for sensitive and selective detection of pollutants in real samples and test papers. Following excitation, PLNPs could store absorbed light energy and release it in the form of luminescence. Compared with a fluorescence-based technique, a PLNPs-based measurement could effectively avoid background interference. Under optimal conditions, the limit of detection for TNP was found to be 10 nM, while for an antibiotic it was 5 nM. The nanoprobe was successfully applied for the detection of pollutants in real samples including milk and Dianchi Lake water samples. Due to the long-lasting afterglow nature of PLNPs, the signal-to-noise ratio could be greatly increased in complex real samples. By hand-writing with TNP solution as ink on filter paper, the photoluminescence (PL) of the part stained with TNP was immediately quenched. Moreover, after direct exposure under a UV lamp for 10 min and without further excitation, the luminescence of the test paper was investigated to avoid interferents. This PLNP material could be potentially employed as a multi-responsive luminescent sensor. In addition, these easy-to-use visual techniques could provide a powerful tool for a convenient POC assay of organic pollutants.
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DNA molecular machines show great promise in fields such as biomarker discovery and biological activity regulation, but operating DNA machines with specific functions within living systems remains extremely challenging. Although DNA machines have been engineered with exact molecular-level specifications, some intrinsic imperfections such as poor cell permeation and fragility in complex cytoplasmic milieu persist due to the well-established character of nucleic acid molecules. To circumvent these problems, we herein report a molecularly engineered, entropy-driven three-dimensional DNA amplifier (EDTD) that can operate inside living cells in response to a specific mRNA target. In particular, mRNA target/EDTD interaction can specifically initiate an autonomous DNA circuit inside living cells owing to the exclusive entropy-driven force, thus providing enormous signal amplification for ultrasensitive detection of the mRNA. Moreover, owing to molecular engineering of a unique DNA tetrahedral framework into the DNA amplifier, EDTD exhibits significantly enhanced biostability and cellular uptake efficiency, which are prerequisites for DNA machines used for in vivo applications. This programmable DNA machine presents a simple and modular amplification mechanism for the detection of intracellular biomarkers. Moreover, this study provides a potentially valuable molecular tool for understanding the chemistry of cellular systems and offers a design blueprint for further expansion of DNA nanotechnology in living systems.
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DNA/química , RNA Mensageiro/química , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Hep G2 , Humanos , Imagem Óptica , Relação Estrutura-Atividade , TermodinâmicaRESUMO
Accurate detection and imaging of tumor-related mRNA in living cells hold great promise for early cancer detection. However, currently, most probes designed to image intracellular mRNA confront intrinsic interferences arising from complex biological matrices and resulting in inevitable false-positive signals. To circumvent this problem, an intracellular DNA nanoprobe, termed DNA tetrahedron nanotweezer (DTNT), was developed to reliably image tumor-related mRNA in living cells based on the FRET (fluorescence resonance energy transfer) "off" to "on" signal readout mode. DTNT was self-assembled from four single-stranded DNAs. In the absence of target mRNA, the respectively labeled donor and acceptor fluorophores are separated, thus inducing low FRET efficiency. However, in the presence of target mRNA, DTNT alters its structure from the open to closed state, thus bringing the dual fluorophores into close proximity for high FRET efficiency. The DTNT exhibited high cellular permeability, fast response and excellent biocompatibility. Moreover, intracellular imaging experiments showed that DTNT could effectively distinguish cancer cells from normal cells and, moreover, distinguish among changes of mRNA expression levels in living cells. The DTNT nanoprobe also exhibits minimal effect of probe concentration, distribution and laser power as other ratiometric probe. More importantly, as a result of the FRET "off" to "on" signal readout mode, the DTNT nanoprobe almost entirely avoids false-positive signals due to intrinsic interferences, such as nuclease digestion, protein binding and thermodynamic fluctuations in complex biological matrices. This design blueprint can be applied to the development of powerful DNA nanomachines for biomedical research and clinical early diagnosis.
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DNA de Cadeia Simples/química , Transferência Ressonante de Energia de Fluorescência/métodos , Nanoestruturas/química , Neoplasias/química , RNA Mensageiro/análise , Biomarcadores Tumorais/análise , Linhagem Celular Tumoral , Sobrevivência Celular , Corantes Fluorescentes/química , Humanos , Limite de Detecção , Neoplasias/diagnóstico , Imagem Óptica/métodos , TermodinâmicaRESUMO
The monitoring of K+ is very important and emergency because of their unique relationship in various disease diagnosis and treatment. G-quadruplex analogue is a classical recognition unit for K+ detection and has been widely applied in K+ relevant research. Common fluorescent dyes were employed for design of G-quadruplex structure-based K+ probes which suffered from the aggregation-caused quenching effect, and possibly limited the biological applications in living systems. Herein, we report an aggregation-induced emission (AIE) effect-based fluorescent probe for cellular K+ analysis and imaging. Benefitting from the K+ triggered AIE phenomenon, the designed TPE derivative modified guanine (G)-rich oligonucleotide fluorescent probe (TPE-oligonucleotide probe) exhibits high sensitivity (â¼10-fold higher than most reported G-quadruplex-based probes) with extended photostability which facilitates the prolonged fluorescence observations of K+ in living cells. On the basis of these advantages, the TPE-oligonucleotide probe serves as a promising candidate for the functional study and analysis of K+.
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DNA/química , Corantes Fluorescentes/química , Oligonucleotídeos/química , Imagem Óptica/métodos , Potássio/análise , Estilbenos/química , Técnicas Biossensoriais/métodos , Células HeLa , Humanos , Potássio/química , Espectrometria de Fluorescência/métodosRESUMO
A universal aptameric system based on the taking advantage of double-stranded DNA/perylene diimide (dsDNA/PDI) as the signal probe was developed for multiplexed detection of small molecules. Aptamers are single-stranded DNA or RNA oligonucleotides which are selected in vitro by a process known as systematic evolution of ligands by exponential enrichment. In this work, we synthesized a new kind of PDI and reported this aggregated PDI could quench the double-stranded DNA (dsDNA)-labeled fluorophores with a high quenching efficiency. The quenching efficiencies on the fluorescence of FAM, TAMRA and Cy5 could reach to 98.3%±0.9%, 97.2%±0.6% and 98.1%±1.1%, respectively. This broad-spectrum quencher was then adopted to construct a multicolor biosensor via a label-free approach. A structure-switching-triggered enzymatic recycling amplification was employed for signal amplification. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity towards small analytes. For other targets, changing the corresponding aptamer can achieve the goal. The quencher did not interfere with the catalytic activity of nuclease. The biosensor could be manipulated with similar sensitivity no matter in pre-addition or post-addition manner. Moreover, simultaneous and multiplexed analysis of several small molecules in homogeneous solution was achieved, demonstrating its potential application in the rapid screening of multiple biotargets.
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Adenosina/análise , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Cocaína/análise , Inibidores da Captação de Dopamina/análise , Corantes Fluorescentes/química , Imidas/química , Perileno/análogos & derivados , Adenosina/sangue , Carbocianinas/química , Cocaína/sangue , Inibidores da Captação de Dopamina/sangue , Humanos , Limite de Detecção , Perileno/química , Espectrometria de Fluorescência/métodosRESUMO
Hydrogen sulfide (H2S), one of the biologically important gaseous signal molecules, plays an essential role in diverse normal biochemical functions and pathological processes. Herein, an efficient two-photon in and near-infrared out mitochondria-targeting dye has been designed, synthesized and characterized. It is easily synthesized by the condensation reaction (CËC) of 4-hydroxybenzaldehyde and 6-(diethylamino)-1,2,3,4-tetrahydroxanthylium (mitochondria-targeting), which possesses large two-photon action absorption cross-section ~160g and high fluorescence quantum yield ~0.15. Encouraged by the results, we proceeded to conjugate this new dye with a H2S recognition moiety (4-dinitrobenzene-ether, DNB), on the basis of the intramolecular charge transfer (ICT) strategy, to construct a novel H2S fluorescent probe (TP-NIR-HS), which shows a targeting ability with high sensitivity and selectivity, and low cytotoxicity. This new probe was then applied for two-photon imaging of living cells and tissues and showed high imaging resolution and a deep-tissue imaging depth of ~350µm, thus demonstrating its practical application in biological systems, and providing a valuable theoretical basis and technical support for the study of physiological and pathological functions of H2S.
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Corantes Fluorescentes/química , Gasotransmissores/análise , Sulfeto de Hidrogênio/análise , Mitocôndrias/química , Imagem Óptica/métodos , Animais , Técnicas Biossensoriais/métodos , Linhagem Celular , Humanos , Raios Infravermelhos , Camundongos Nus , Microscopia de Fluorescência/métodos , Espectrometria de Fluorescência/métodosRESUMO
Hypochlorite anion, one of the biologically important reactive oxygen species, plays an essential role in diverse normal biochemical functions and abnormal pathological processes. Herein, an efficient high-resolution mitochondria-targeting ratiometric fluorescent probe for hypochlorous acid detection has been designed, synthesized and characterized. It is easily synthesized by the condensation reaction (CC) of a 2-(2-hydroxyphenyl) quinazolin-4(3H)-one fluorophore and a cyanine group (mitochondria-targeting), which made the whole molecular a large Stokes shift (210nm) and the two well-resolved emission peaks separated by 140nm. As a result, it is considered as a good candidate for high resolution hypochlorous acid imaging in live cells. The ratiometric fluorescent probe exhibited outstanding features of high sensitivity, high selectivity, rapid response time (within 50s), and excellent mitochondria-targeting ability. Moreover, the probe can also be successfully applied to imaging endogenously hypochlorous acid in the mitochondria of living cells with low cytotoxicity, and high resolution.
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Carbocianinas/química , Corantes Fluorescentes/química , Ácido Hipocloroso/análise , Mitocôndrias/química , Quinazolinonas/química , Linhagem Celular , Humanos , Microscopia de Fluorescência/métodos , Mitocôndrias/ultraestrutura , Imagem Óptica/métodos , Espectrometria de Fluorescência/métodosRESUMO
Conjugation of DNAs to defined locations on a protein surface will offer powerful tools for positioning functional groups and molecules in biological and biomedical studies. However, tagging protein with DNA is challenging in physiological environments, which requires a bioorthogonal approach. Here we report a chemical solution to selectively conjugate DNA aptamer with a protein by protein-aptamer template (PAT)-directed reactions. Since protein-aptamer interactions are bioorthogonal, we exploit PAT as a unique platform for specific DNA-protein cross-linking. We develop a series of modified oligonucleotides for PAT-directed reactions and screen out F-carboxyl as a suitable functionality for selective and site-specific conjugation. The functionality is incorporated into aptamers by our F-carboxyl phosphoramidite with easy synthesis. We also demonstrate the necessity of a linker between the reactive functionality and the aptamer sequences.
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Lysosomes are acidic organelles (approximately pH 4.5-5.5) and tracking the changes in lysosomal pH is of great biological importance. To address this issue, quite a few of fluorescent probes have been developed. However, few of these probes can realize the tracking of dynamic changes in lysosomal pH. Herein, we report a new lysosome-targeted ratiometric fluorescent probe (FR-Lys) by hybridizing morpholine with a xanthane derivative and an o-hydroxy benzoxazole group. In this probe, the morpholine group serves as a targeting unit for lysosome, the xanthane derivative exhibits a pH-modulated open/close reaction of the spirocycle, while the o-hydroxy benzoxazole moiety shows a pH modulated excited-state intramolecular proton transfer (ESIPT) process. Such a design affords the probe a ratiometric fluorescence response towards pH with pH values ranging from 4.0 to 6.3. The response of the probe to pH was fast and reversible with high selectivity. Moreover, this probe possesses further advantages such as easy synthesis, high photostability and low cytotoxicity. These features are favorable for tracking dynamic pH changes in biosystems. It was then applied for dynamic imaging pH changes in lysosomes with satisfactory results.
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Benzoxazóis/química , Corantes Fluorescentes/química , Lisossomos/química , Técnicas de Sonda Molecular , Prótons , Espectrometria de Fluorescência/métodos , Antimaláricos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Cloroquina/farmacologia , Diagnóstico por Imagem , Células HeLa , Humanos , Concentração de Íons de HidrogênioRESUMO
Photoactivatable probe-based fluorescent imaging has become an efficient and attractive technique for spatiotemporal microscopic studies of biological events. However, almost all previously reported photoactivatable organic probes have been based on hydrosoluble precursors, which have produced water-soluble active fluorophores able to readily diffuse away from the photocleavage site, thereby dramatically reducing spatial resolution. Hydroxyphenylquinazolinone (HPQ), a small organic dye known for its classic luminescence mechanism through excited-state intramolecular proton transfer (ESIPT), shows strong light emission in the solid state, but no emission in solution. In this work, HPQ was employed as a precursor to develop a localizable, photoactivatable two-photon probe (PHPQ) for spatiotemporal bioimaging applications. After photocleavage, PHPQ releases a precipitating HPQ fluorophore which shows both one-photon and two-photon excited yellow-green fluorescence, thereby producing a localizable fluorescence signal that affords high spatial resolution for bioimaging, with more than 200-fold one-photon and 150-fold two-photon fluorescence enhancement.