RESUMEN
Endogenous sulfur dioxide (SO2), as a gas signal molecule, has a certain physiological functions. Understanding the role of endogenous SO2 in human physiology and pathology is of great significance to the biological characteristics of SO2,which bring challenges to develop fluorescent probes with excellent performance. Herein, we rationally designed and constructed a novel near-infrared bioprobe benzaldehyde-benzopyrylium (BBp) by employing the nucleophilic addition benzopyrylium perchlorate fluorophore and benzaldehyde moiety by means of C = C/C = O group that serves as both fluorescence reporting unit. Probe BBp exhibit excellent sensing performance with fluorescent "On - Off"rapid response (100 s) and long-wavelength emission (670 nm). With the treatment of HSO3-, the color of BBp solution obviously varies from purple to colorless, and the fluorescent color varies from red to colorless. By the fluorescence and colorimetric changes, probe BBp was capable of sensitive determination HSO3- with low limits of detection (LOD) of 0.43 µM, realizing visual quantitative monitoring SO2 derivative levels. Due to the low phototoxicity and good biocompatibility, it was successfully applied to monitor SO2 derivatives and fluorescence imaging in HepG2 and HeLa living cells. Hopefully, this work supplies a new strategy for designing NIR fluorescent probes for quantitative determination SO2 derivatives in biological samples.
Asunto(s)
Benzaldehídos , Colorantes Fluorescentes , Humanos , Percloratos , Células HeLa , MitocondriasRESUMEN
Astragalus membranaceus is a traditional Chinese medicine derived from the roots of Astragalus membranaceus (Fisch.) Bge., which has the same medicinal and edible uses in China. It is also widely used in daily food, and its pharmacological effects mainly include antioxidant effects, vascular softening effects, etc. Currently, it is increasingly widely used in the prevention of hypertension, cerebral ischemia, and stroke in China. Formononetin and its glucopyranoside (ononin) are both important components of Astragalus membranaceuss and may play important roles in the treatment of cardiovascular diseases (CVDs). This study conducted metabolic studies using formononectin and its glucopyranoside (ononin), including a combination of the in vitro metabolism of Formonetin using rat liver S9 and the in vivo metabolism of ononin administered orally to rats. Five metabolites (Sm2, 7, 9, 10, and 12) were obtained from the solution incubated with formononetin and rat hepatic S9 fraction using chromatographic methods. The structures of the five metabolites were elucidated as (Sm2)6,7,4'-trihydroxy-isoflavonoid; (Sm7)7,4'-dihydroxy-isoflavonoid; (Sm9)7,8,4'-trihydroxy-isoflavonoid; (Sm10)7,8,-dihydroxy-4'-methoxy-isoflavonoid; and (Sm12)6,7-dihydroxy-4'-methoxy- isoflavonoid on the basis of UV, NMR, and MS data. Totally, 14 metabolites were identified via HPLC-DAD-ESI-IT-TOF-MSn analysis, from which the formononetin was incubated with rat hepatic S9 fraction, and the main metabolic pathways were hydroxylation, demethylation, and glycosylation. Then, 21 metabolites were identified via HPLC-DAD-ESI-IT-TOF-MSn analysis from the urine samples from SD rats to which ononin was orally administered, and the main metabolic pathways were glucuronidation, hydroxylation, demethylation, and sulfonation. The main difference between the in vitro metabolism of formononetin and the in vivo metabolism of ononin is that ononin undergoes deglycemic transformation into Formonetin in the rat intestine, while Formonetin is absorbed into the bloodstream for metabolism, and the metabolic products also produce combined metabolites during in vivo metabolism. The six metabolites obtained from the aforementioned separation indicate the primary forms of formononetin metabolism, and due to their higher contents of similar isoflavone metabolites, they are considered the main active compounds that are responsible for pharmacological effects. To investigate the metabolites of the active ingredients of formononetin in the rat liver S9 system, network pharmacology was used to evaluate the cardiovascular disease (CVD) activities of the six primary metabolites that were structurally identified. Additionally, the macromolecular docking results of six main components and two core targets (HSP90AA1 and SRC) related to CVD showed that formononetin and its main metabolites, Sm10 and Sm12, may have roles in CVD treatment due to their strong binding activities with the HSP90AA1 receptor, while the Sm7 metabolite may have a role in CVD treatment due to its strong binding activity with the SRC receptor.
Asunto(s)
Enfermedades Cardiovasculares , Medicamentos Herbarios Chinos , Isoflavonas , Ratas , Animales , Ratas Sprague-Dawley , Medicamentos Herbarios Chinos/química , Farmacología en Red , Isoflavonas/química , Cromatografía Líquida de Alta Presión/métodos , Hígado/metabolismoRESUMEN
To date, the effective discrimination of anionic sulfonate surfactants with tiny differences in structure, considered as environmentally noxious xenobiotics, is still a challenge for traditional analytical techniques. Fortunately, a sensor array becomes the best choice for recognizing targets with similar structures or physical/chemical properties by virtue of principal component analysis (PCA, a statistical technique). Herein, because of the beneficial construction of the statistical strategy and use of two types of luminescent metal-organic frameworks (LMOFs, NH2-UiO-66 and NH2-MIL-88) as sensing elements, high-throughput discrimination and detection of five anionic sulfonate surfactants and their mixtures are nicely realized for the first time. Significantly, the stacking interaction of aromatic rings and dynamic quenching play essential roles in the generation of diverse fluorescence responses and unique fingerprint maps for individual anionic sulfonate surfactants. Moreover, the mixtures of anionic sulfonate surfactants are also satisfactorily distinguished in environmental water samples, demonstrating the practicability of the sensor array. On the basis of the PCA method, this strategy converts general fluorescence signals into unique optical fingerprints of individual analytes, providing a new opportunity for the application of LMOFs in the field of analytes recognition.
RESUMEN
Developing a multifunctional platform for the selective detection and effective removal of toxic ions is a major challenge when addressing heavy metal contamination in environmental science. Herein, novel nonconjugated polymer nanoparticles (PNPs) called mercaptosuccinic acid-thiosemicarbazide PNPs (MT-PNPs) with appealing fluorescence and stability are synthesized via facile one-step hydrothermal treatment for attractive sensing and simultaneous removal of mercury(II). Interestingly, aggregation-induced fluorescence switch-off and scattering enhancement are found upon the addition of Hg2+, rendering MT-PNPs as a ratiometric sensor for selective and accurate Hg2+ monitoring. A wide linear range (0.1-1471 µM) and a low detection limit (95 nM) are obtained. This dual-signal opposite responses triggered by Hg2+ originate from the formation of MT-PNP-Hg2+ congeries via the multisite binding between S,N,O-containing groups of MT-PNPs and mercury. Meanwhile, target-induced aggregation renders an effective Hg2+ separation from contaminative aqueous media by MT-PNPs, which exhibits a satisfactory absorption efficiency of 90.42% within 50 min. Upon the simple Na2S treatment, the MT-PNPs can be regenerated and reused. This work thus delivers an applicable method for the ratiometric detection and effective removal of mercury with the novel nonconjugated PNPs, offering potential in tackling the problem of heavy metal ion pollution for environmental monitoring and remediation.
Asunto(s)
Mercurio , Nanopartículas , Colorantes Fluorescentes , Iones , Límite de Detección , PolímerosRESUMEN
In order to detect ascorbic acid (AA) sensitively, a dual-signal optical sensor of a nanosystem with carbon dots (CDs)/MnO2 nanosheets based on fluorescence and second-order scattering (SOS) has been constructed. Here, MnO2 nanosheets, both as a fluorescence quencher and signal transducer of SOS, quench the blue fluorescence of CDs by an inner filter effect. Under the excitation of 300 nm, the nanosystem shows a fluorescence emission peak at 405 nm and a SOS peak at 610 nm, respectively. With the increase of AA , the lamellar structure of MnO2 nanosheets is etched into a smaller nanostructure, causing a decrease of the fluorescence recovery of CDs (405 nm) and decrease of the SOS signal of the MnO2 nanosheets (610 nm). According to the simultaneous changes of fluorescence and SOS signals, a dual-signal optical sensor toward AA is successfully constructed. Satisfactorily, the optical sensor for AA detection shows a detection limit of 88 and 105 nM for fluorescence and SOS, respectively. The practical application of the designed sensor is verified through the detection of AA content in vitamin C tablets, and satisfactory results are obtained Graphical Abstract A dual-signal sensor of fluorescence (FL) and second-order scattering (SOS) based on the carbon dot (CD) and MnO2 nanosheet system for ascorbic acid (AA) detection is constructed, in which CDs are used for the FL mode and MnO2 nanosheets are used for the SOS mode.
Asunto(s)
Ácido Ascórbico/análisis , Colorantes Fluorescentes/química , Compuestos de Manganeso/química , Óxidos/química , Puntos Cuánticos/química , Espectrometría de Fluorescencia/métodos , Ácido Ascórbico/química , Carbono/química , Fluorescencia , Límite de Detección , Oxidación-Reducción , Comprimidos/análisisRESUMEN
Rapid and sensitive detection of surfactants has attracted more and more attention since surfactants not only cause water pollution but also affect the health of human beings. Luminescent metal-organic frameworks combining unique optical property and inherent permanent porosity for guest-host encapsulation are widely used in fluorescence detection. Here we report a ratiometric fluorescent probe (denoted as UiO-66-NH2@PB) based on a Zr-based metal-organic framework (UiO-66-NH2) and a fluorescent dye, phloxine B (PB), for visual and fluorescent determination of cationic surfactants (cetyltrimethylammonium bromide; CTAB). The intensity ratio of dual-emission sensor exhibits a linear response to the CTAB concentrations of 0.1-17 µM and obtains a low detection limit (0.074 µM). Moreover, this method has been successfully utilized to monitor CTAB in the environmental water samples with satisfied recoveries. Importantly, this work provides a new insight into developing smartphone-based sensor to realize a rapid, on-site visual and quantification-based detection of CTAB.
RESUMEN
Bisulfite and hydrazine are harmful to the environment safety and human health. Therefore, it is of great value to develop a smart fluorescent probe with high selectivity for detection of bisulfite and hydrazine. In our report, a dual-response fluorescent probe EDBI with high selectivity, rapid response, and low detection limit for discriminative determination HSO3- and N2H4 was exploited. The probe EDBI is capable of distinctive sensing HSO3- and N2H4 based on nucleophilic addition reactions by taking advantage of ratiometric fluorescence and fluorescence "on-off" mode, respectively. The dual-responses behaviors of probe EDBI toward HSO3- and N2H4 were attribute to different reaction sites, which it has been confirmed by HRMS. More importantly, cytotoxicity experiment authenticated that probe possesses low toxicity and good penetration. The probe EDBI with excellent performance, it was successfully employed to distinguishable sense HSO3- and N2H4 in living cells by diverse channel patterns. Therefore, this simple dual-response fluorescence probe is expected to be used for real-time monitoring bisulfite and hydrazine in biological samples.
Asunto(s)
Colorantes Fluorescentes , Sulfitos , Diagnóstico por Imagen , Humanos , Hidrazinas , Espectrometría de FluorescenciaRESUMEN
A new type of dye with advantages of high selectivity and sensitivity is formed by using the strategy of hybridization between the luminescent unit and recognition unit. Based on this strategy, we exploit a novel dye bonding the benzopyrylium salt as a luminescent unit and phenylboronate group as a response site, which is served as a fluorescent probe 1 for specific recognition of hydrogen peroxide in biological application. Probe 1 employs a unique recognition switch, phenylboronate unit, to"turn-on"a highly specific and rapid fluorescence response toward hydrogen peroxide combined with the 1,6-rearrangement elimination reaction strategy. Meanwhile, probe 1 has the ability to glucose assay by taking advantage of glucose oxidase/glucose enzymatic reaction. What's more, the probe 1 is capable of tracking endogenous hydrogen peroxide in living cells and intracellular imaging. Therefore, the newly developed bioprobe 1 is expected to be used to monitor hydrogen peroxide and glucose levels in complex organisms.
Asunto(s)
Técnicas Biosensibles , Colorantes Fluorescentes , Glucosa , Glucosa Oxidasa , Peróxido de HidrógenoRESUMEN
Esterase activity is often used as an index to evaluate the health status of cells and plays an important role in cell metabolism and apoptosis. Herein, we develop two fluorescent probes for visual biosensing of esterase activity and imaging in living cells. In vitro, after the introduction of esterase, enzymolysis destroys the ester bond of the probe, causing the fluorescent color of probe changes from yellow to red, thus realizing the visual strategy for determination of esterase activity, with high sensitivity and selectivity. Especially, probe VA, 2-(4-acetoxystyryl)-3-ethyl-1,1-dimethyl- 1H-benzo[e]indol-3-ium, exhibits higher sensitivity with a lower detection limit (up to 7.15 × 10-6 U/mL). In the cell experiment, the fluorescent probe VA also shows good biocompatibility and high spatial resolution, and is successfully applied to the intracellular fluorescent imaging and biosensing of esterase in living cells. More importantly, the probe VA can judge the unhealthy state of H2O2-induced HeLa cells using dual-fluorescence signals. The results confirm that the fluorescence method is a reliable tool for detecting endogenous esterase in living biological system.
Asunto(s)
Colorantes Fluorescentes , Peróxido de Hidrógeno , Esterasas , Células HeLa , Humanos , Espectrometría de FluorescenciaRESUMEN
The aim of this study was to isolate a mould from the surface of manganese slag which had strong resistance and high adsorption of Mn(2 + ), and to determine the effects of initial Mn(2 + ) concentration, incubation temperature, rotation speed and inoculation amount on adsorption of Mn(2 + ) from manganese waste water solution. The result showed that a mould (A5) which was isolated from manganese slag had the adsorption rate of Mn(2 + ) to 97.5% at the initial pH value 6, inoculation amount 2%, rotation speed 150 r/min, a concentration of Mn(2 + ) 500 mg/L, and a temperature of 28 degrees C cultivated for 50 h. As there is no research on adsorption of Mn(2 + ) from manganese waste water by fungi before, this research showed a theoretical guidance on this field.
Asunto(s)
Hongos/metabolismo , Manganeso/química , Eliminación de Residuos Líquidos/métodos , Biodegradación Ambiental , Mentón , Residuos Industriales , Minería , Factores de TiempoRESUMEN
Herein, an innovative photocathodic enzymatic biosensor is proposed with poly {4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]-benzo[1,2-b:4,5-b']dithiophene-2,6-diyl-alt-3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophene-4,6-diyl} (PTB7-Th) as donor-acceptor-type photoactive material and three-dimensional (3D) polyaniline hydrogels (PAniHs) as both electron transfer layer and biomolecule carrier. Based on the enhancement effect of PAniHs on the charge separation and electron transfer of PTB7-Th and the competitive consumption of dissolved oxygen (O2) between the xanthine oxidase (XOD)-guanine catalytic reaction and O2-sensitive PTB7-Th/PAniHs, the proposed photocathodic enzymatic biosensor has been demonstrated to detect guanine with the advantages of low limit of detection (0.02 µM), wide linear range (from 0.1 to 80 µM), simple and convenient preparation process, satisfactory stability, and photochemical signal amplification independent of any exogenous electron donor/acceptor or sensitizer. Remarkably, the proposed photocathodic enzymatic biosensor can not only be extended to other aerobic enzymatic bioanalyses, but also pave a horizon for the application of environmentally friendly conductive hydrogel materials in photoelectrochemical bioanalysis.
Asunto(s)
Compuestos de Anilina , Técnicas Biosensibles/métodos , Pruebas de Enzimas/métodos , Hidrogeles , Procesos Fotoquímicos , Compuestos de Anilina/química , Biocatálisis , Técnicas Biosensibles/instrumentación , Pruebas de Enzimas/instrumentación , Pruebas de Enzimas/normas , Hidrogeles/química , Estructura Molecular , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Análisis EspectralRESUMEN
Intracellular pH level plays an important role in physiological and pathological processes. The development of nanoprobes for detecting in vivo pH levels is especially important for early diagnosis of disease. Therefore, we develop a hydrophilic carbon points (CDs) using quercetin and ethylenediamine as precursors to monitor intracellular pH. Under optimized conditions, the prepared CDs not only have uniform particle size and morphology, but also possess strong green fluorescence, photostability, and photoreversibility in water medium. The CDs exhibit pH-sensitive fluorescence effect under acidic and alkaline conditions, which is used to achieve "off-on-off" detection pH (from 3.5 to 13.5). Meanwhile, the pH-dependent mechanism is further investigated and explained, which is the fluorescence quenching caused by the pH-induced aggregation. Based on the pH-sensitive characteristics of CDs, it has been applied to the detection of aspartic acid and glutamic acid. More importantly, when applied to live cells, the pH-probe exhibits low cytotoxicity and high sensitivity, and is successfully used in intracellular pH fluorescence imaging. Consequently, this nanoprobe is expected to be used for real-time monitoring of intracellular pH level.
Asunto(s)
Aminoácidos Acídicos/análisis , Carbono/química , Puntos Cuánticos/química , Fluorescencia , Células Hep G2 , Humanos , Concentración de Iones de Hidrógeno , Interacciones Hidrofóbicas e Hidrofílicas , Punto Isoeléctrico , Tamaño de la Partícula , Puntos Cuánticos/toxicidad , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta , Espectrometría Raman , Difracción de Rayos XRESUMEN
Development of selective and sensitive methods for on-site assay of tetracycline (TC) is of great significance for public health and food safety. Herein, a valid ratiometric fluorescence strategy using g-C3N4 nanosheets coupled with Eu3+ is designed for the assay of TC. In this strategy, both Eu3+ and g-C3N4 nanosheets serve as the recognition units of TC. The blue fluorescence of g-C3N4 nanosheets can be quenched by TC via the inner filter effect (IFE); meanwhile, the red fluorescence of Eu3+ can be enhanced by TC through the antenna effect (AE). The synergistic effect of AE and IFE caused by TC makes the developed ratiometric fluorescent sensor display a wide linear range for TC from 0.25 to 80 µM with a detection limit of 6.5 nM and a significant fluorescence color evolution from blue to red. Given its simplicity, free-label, excellent selectivity, high sensitivity, and recognizable color change, point-of-care testing systems, including smartphones and test paper-based assays, are developed for the visual sensing of TC. The integration of smartphones and test paper on a ratiometric fluorescent sensor greatly reduces the detection cost and time, providing a promising method for the qualitative discernment and semi-quantitative assay of TC on-site. Moreover, the potential application of the approach is also verified by detecting TC in milk.
Asunto(s)
Técnicas Biosensibles , Colorantes Fluorescentes/química , Papel , Teléfono Inteligente , Tetraciclina/análisis , Animales , Europio/química , Leche/química , Conformación Molecular , Nitrilos/química , Tamaño de la Partícula , Espectrometría de Fluorescencia , Propiedades de SuperficieRESUMEN
OBJECTIVE: To test the hypothesis that the inhibition of endoplasmic reticulum (ER) stress-induced apoptosis in oxidized low-density lipoproteins (ox-LDL)-induced human aortic-vascular smooth muscle cells (HA-VSMCs) was associated with suppression of the protein kinase RNA-like ER kinase (PERK)-eukaryotic translation initiation factor 2α (eIF2α)-activating transcription factor 4 (ATF4)-CCAAT/enhancer binding protein homologous protein (CHOP) signaling pathway by Pollen Typhae total flavone (PTF). METHODS: Primary HA-VSMCs were cultured and identified. The cultured HA-VSMCs were randomized into 5 groups, including a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an HPTF group (70 µg/mL high ox-LDL+500 µg/mL PTF), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), and a LPTF group (70 µg/mL high ox-LDL+100 µg/mL PTF) in the first part; and a normal control group, an ox-LDL group (70 µg/mL high ox-LDL), an MPTF group (70 µg/mL high ox-LDL+250 µg/mL PTF), a shRNA group (transducted with PERK shRNA lentiviral particles), a scramble shRNA group (transducted with control shRNA lentiviral particles), an MPTF+ox-LDL+shRNA group (250 µg/mL PTF+70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) and an ox-LDL+shRNA group (70 µg/mL high ox-LDL+PERK shRNA lentiviral particles) in the second part. The protein expression levels of ER-associated apoptosis proteins were detected by Western blot, and their mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was applied to test cell viability, and the level of apoptosis was monitored by flow cytometry. RESULTS: The MTT assay and flow cytometry showed that the ox-LDL group had a significant increase in apoptosis, which was attenuated in PTF treatment groups and shRNA groups. Moreover, the ox-LDL group had increased protein and mRNA levels of binding immunoglobulin protein and ER-associated apoptosis proteins, such as PERK, eIF2α, ATF4 and CHOP, which were attenuated in PTF treatment groups and shRNA groups. CONCLUSIONS: The apoptosis induced by ox-LDL had a strong relation to ER stress. The protective effect of PTF on ER stressinduced apoptosis was associated with inhibition of the PERK-eIF2α-ATF4-CHOP pathway, which might be a potential therapeutic strategy for enhancing the stability of atherosclerotic plaques.
Asunto(s)
Apoptosis/efectos de los fármacos , Regulación hacia Abajo , Medicamentos Herbarios Chinos/farmacología , Estrés del Retículo Endoplásmico/efectos de los fármacos , Flavonas/farmacología , Músculo Liso Vascular/patología , Miocitos del Músculo Liso/patología , Transducción de Señal , Factor de Transcripción Activador 4/metabolismo , Aorta/patología , Proliferación Celular/efectos de los fármacos , Factor 2 Eucariótico de Iniciación/metabolismo , Humanos , Miocitos del Músculo Liso/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal/efectos de los fármacos , Factor de Transcripción CHOP/metabolismo , eIF-2 Quinasa/metabolismoRESUMEN
Copper is an essential mineral nutrient for the human body. However, excessive levels of copper accumulated in the body can cause some diseases. Therefore, it is great significant to establish a sensitive bioprobe to recognize copper ions (Cu2+) in vivo. In our work, nitrogen-doped carbon dots (N-CDs) and gold nanoclusters (Au NCs) are selected as luminescent nanomaterials and the Au NCs/N-â¯CDs nanohybrids is successfully synthesized by coupling method. The Au NCs/N-CDs exhibited characteristic dual-emission peaks at 450 and 620â¯nm when excited by a single-wavelength of 380â¯nm. When different amounts of Cu2+ are introduced, the fluorescence intensity of the Au NCs is gradually weakened and fluorescence intensity of the N-CDs is almost unchanged, which can facilitate the visual detection of Cu2+. The Au NCs/N-CDs nanohybrid possesses good selectivity to Cu2+ with a limit of detection (LOD) is 3.5⯵M and linear detection range of 10-150⯵M. Visualization detection of Cu2+ is implemented by using nanoprobe in water samples. Furthermore, the ratiometric nanoprobe is utilized to the toxicity test of liver cancer cells, indicating excellent biocompatibility and low toxicity. This nanoprobe has been used to the intracellular fluorescence imaging. Moreover, this method is expected to be used to monitor the changes of Cu2+ concentration in hepatocytes.
Asunto(s)
Cobre/análisis , Espacio Intracelular/metabolismo , Sondas Moleculares/química , Nanopartículas/química , Imagen Óptica , Carbono/química , Oro/química , Células Hep G2 , Humanos , Nitrógeno/química , Espectrometría de Fluorescencia , Espectrofotometría Ultravioleta , Agua/químicaRESUMEN
Accumulating evidence suggests that peroxiredoxins (Prxs) eliminate excessive cellular H2O2 and are important factors in redox signaling pathways. In this study, we cloned the full-length cDNAs and genomic sequences of Prx3 and Prx4 from common carp. The common carp Prx3 and Prx4 open reading frames were 753 base pairs (bp) and 783bp in length, respectively, and contained seven exons and six introns. Multiple sequence alignment and phylogenetic analyses revealed that the common carp Prx1-4 proteins share high identities and similar characteristics with other known animal Prxs. Prx3 and Prx4 mRNA were constitutively expressed in all tissues, and the highest Prx3 and Prx4 transcript abundances occurred in head kidney. Although the highest Prx4 protein and mRNA expression were also observed in head kidney, many differences were detected between Prx4 mRNA and protein expression levels in other tissues. Prx3 expression increased significantly in the head kidney 12h after an Aeromonas hydrophila challenge. The A. hydrophila challenge upregulated Prx3 mRNA expression in liver and spleen, increased Prx4 mRNA expression levels in liver and spleen excluding at 36h in spleen, but decreased Prx4 mRNA expression level in the head kidney. The mature Prx4 peptide was recombinantly expressed and purified using Dextrin Beads 6FF and it exhibited thioredoxin (Trx)-dependent peroxidase activity. These data suggest that Prx3 and Prx4 are constitutive and inducible proteins that might play important roles in innate immune function. The Trx-dependent peroxidase activity analysis of recombinant Prx4 further verified the important role of Prxs in the redox system of fish.
Asunto(s)
Infecciones Bacterianas/inmunología , Carpas/inmunología , Cisteína , Inmunidad Activa , Peroxiredoxina III/inmunología , Peroxirredoxinas/inmunología , Animales , Proteínas de Arabidopsis/genética , Carpas/genética , Clonación Molecular , Cisteína/química , Peroxidasas/genética , Peroxiredoxina III/química , Peroxiredoxina III/genética , Peroxirredoxinas/química , Peroxirredoxinas/genética , FilogeniaRESUMEN
A photoresponsive hollow molecularly imprinted polymer (PHMIP) was fabricated for photoresponsive recognition and determination of trace bisphenol A (BPA) in aqueous media using a water-soluble azo compound as the functional monomer. The PHMIP was prepared on sacrificial silica microspheres by surface imprinting and subsequent removal of the silica core. The PHMIP displayed photocontrolled recognition for BPA. SEM, TEM, FT-IR, TGA and N2 adsorption-desorption analyses confirmed successful formation of the hollow structure. The PHMIP displayed higher binding capacity, a larger specific area, and faster mass transfer rate than its corresponding surface molecularly imprinted polymer. The PHMIP was used to determine trace BPA in real samples with a limit of detection of 0.5ppm. For samples spiked at 0-10ppm, the BPA recoveries were in the range of 93.0%-99.0%. This PHMIP-based method provides convenient and inexpensive detection method for trace BPA in environmental samples. This method is especially suitable for determining materials that do not possess specific spectroscopic or luminescent properties.
RESUMEN
A novel photoresponsive molecularly imprinted polymer (MIP) was developed for the selective extraction of guanine from complex samples. The photoresponsive MIP was fabricated using guanine as the template, water-soluble 5-[(4-(methacryloyloxy)phenyl)diazenyl]isophthalic acid as the functional monomer, and water-soluble triethanolamine trimethacrylate as the cross-linker. The MIP displayed good selectivity toward guanine with a dissociation constant of (2.70 ± 0.16) × 10(-5) mol L(-1) in aqueous media. The density of the guanine-specific receptor sites in the MIP material was (4.49 ± 0.22)µmol g(-1). Quantitatively release and uptake of guanine by the MIP occurred with irradiation at 365 and 440 nm, respectively. The MIP could efficiently extract guanine from beer and then release it into aqueous media under photocontrol. This method could be used for selective separation and subsequent determination of a specific analytes from complex samples.