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1.
In the present work, SOD mimetic nanozyme (NACu-Cys) consisting of Cu-Cys complex and nano-albumin (NA) were synthesized. After characterizing the nanozyme, its superoxide dismutase (SOD) behavior was evaluated by inhibition of the pyrogallol autoxidation method. The results revealed that NACu-Cys exhibited SOD mimetic activity with a half inhibition concentration (IC50) value of 7.0 × 103 µM and a turnover number (kcat) of 5.4 × 107 s1. In the next step, this nanozyme was applied as a protective agent against oxidative stress induced by sperm cryopreservation. Increasing the motility, raising the viability and reducing the apoptosis occurred as a result of NACu-Cys additions to human sperm freezing medium. Comparison between the natural SOD and SOD mimic behavior of NACu-Cys revealed that this nanoparticle has the ability to be used as oxidative stress decrescent during cryopreservation process.  相似文献   

2.
A novel flow injection-chemiluminescence (FI–CL) approach is proposed for the assay of pioglitazone hydrochloride (PG-HCl) based on its enhancing influence on the tris(2,2′-bipyridyl)ruthenium(II)–silver(III) complex (Ru(bipy)32+-DPA) CL system in sulfuric acid medium. The possible CL reaction mechanism is discussed with CL and ultraviolet (UV) spectra. The optimum experimental conditions were found as: Ru(bipy)32+, 5.0 × 10−5 M; sulfuric acid, 1.0 × 10−3 M; diperiodatoargentate(III) (DPA), 1.0 × 10−4 M; potassium hydroxide, 1.0 × 10−3 M; flow rate 4.0 ml min−1 for each flow stream and sample loop volume, 180 μl. The CL intensity of PG-HCl was linear in the range of 1.0 × 10−3 to 5.0 mg L−1 (R2 = 0.9998, n = 10) with limit of detection [LOD, signal-to-noise ratio (S/N= 3] of 2.2 × 10−4 mg L−1, limit of quantification (LOQ, S/N = 10) of 6.7 × 10−4 mg L−1, relative standard deviation (RSD) of 1.0 to 3.3% and sampling rate of 106 h−1. The methodology was satisfactorily used to quantify PG-HCl in pharmaceutical tablets with recoveries ranging from 93.17 to 102.77 and RSD from 1.9 to 2.8%.  相似文献   

3.
We studied the postulated involvement of the protein kinase C β1 (PKCβ1) isoform in the regulation of endothelial permeability using human dermal microvascular endothelial cell line (HMEC-1). We overexpressed the recombinant PKCβ1 gene via retroviral-mediated transduction in these cells. PKCβ1 gene transfer was stable, and PKCβ1 protein production was persistent for at least 1 month posttransduction. Addition of 2 × 10−9 M and 2 × 10−8 M phorbol 12-myristate 13-acetate (PMA) to the control (nontransduced) HMEC-1 cells increased the transendothelial 125I-albumin clearance rate (an index of endothelial permeability) from 2.5 ± 0.2 × 10−2 μl/min to 5.4 ± 1.2 × 10−2 μl/min and 16.8 ± 3.1 × 10−2 μl/min, respectively. However, addition of 2 × 10−9 M PMA to PKCβ1-overexpressing HMEC-1 cells produced a maximal increase in the transendothelial 125I-albumin clearance rate of 15.9 ± 2.0 × 10−2 μl/min. Challenge of these cells with 2 × 10 −8 M PMA did not further augment the increase in permeability. Activation with PMA was associated with the translocation of the PKCβ1 from the cytosol to the membrane. These data show that PKCβ1 overexpression augments the increase in endothelial permeability in response to PKC activation, suggesting an important function for the PKCβ1 isoform in the regulation of endothelial barrier. © 1996 Wiley-Liss, Inc.  相似文献   

4.
This study introduces a novel chemiluminescence (CL) approach utilizing FeS2 nanosheets (NSs) catalyzed luminol–O2 CL reaction for the measurement of three pharmaceuticals, namely venlafaxine hydrochloride (VFX), imipramine hydrochloride (IPM), and cefazolin sodium (CEF). The CL method involved the phenomenon of quenching induced by the pharmaceuticals in the CL reaction. To achieve the most quenching efficacy of the pharmaceuticals in the CL reaction, the concentrations of reactants comprising luminol, NaOH, and FeS2 NSs were optimized accordingly. The calibration curves demonstrated exceptional linearity within the concentration range spanning from 4.00 × 10−7 to 1.00 × 10−3 mol L−1, 1.00 × 10−7 to 1.00 × 10−4 mol L−1, and 4.00 × 10−6 to 2.00 × 10−4 mol L−1 with detection limits (3σ) of 3.54 × 10−7, 1.08 × 10−8, and 2.63 × 10−6 mol L−1 for VFX, IPM, and CEF, respectively. This study synthesized FeS2 NSs using a facile hydrothermal approach, and then the synthesized FeS2 NSs were subjected to a comprehensive characterization using a range of spectroscopic methods. The proposed CL method was effective in measuring the aforementioned pharmaceuticals in pharmaceutical formulations as well as different water samples. The mechanism of the CL system has been elucidated.  相似文献   

5.
6.
[22,23-3H2]dihydroazadirachtin was incorporated by Sf9 cells in culture and was bound specifically to the nuclear fraction. The observed association constant of the binding of the radioligand to a purified nuclear fraction was determined to be 0.037 ± 0.008 min 1 using a one-phase exponential association equation, and binding appeared to be to a single population of sites. The binding was essentially irreversible, and the dissociation constant was estimated to be 0.00065 ± 0.00013 min 1. An association rate constant of 7.3 × 106 M 1 min 1 was calculated from these data. Binding was saturable, and the receptor number and affinity were determined as Bmax = 23.87 ± 1.15 pmol/mg protein, Kd = 18.1 ± 2.1 nM. The order of potency of semisynthetic azadirachtin analogues for competition for the binding site was as follows (IC30 in parentheses): azadirachtin (1.55 × 10−8 M) > dihydroazadirachtin (3.16 × 10−8 M) > dansyl dihydroazadirachtin (7.40 × 10−8 M) > DNP-azadirachtin (7.50 × 10−8 M) > biotin dihydroazadirachtin (1.27 × 10−7 M) ≫ 11-methoxy 22,23-dihydroazadirachtin (6.67 × 10−7 M). Arch. Insect Biochem. Physiol. 34:461–473, 1997. © 1997 Wiley-Liss, Inc.  相似文献   

7.
A sensitive fluorescence (FL) technique is proposed for the determination of levofloxacin (LVX). The method is based on the fact that the weak FL signal of the Tb(III)–LVX system is strongly enhanced in the presence of gold nanoparticles. Gold nanoparticles were prepared by the citrate reduction of HAuCl4 and characterized by transmission electron microscopy (TEM). Levofloxacin and Tb(III) ion form a fluorescence complex in aqueous solution, and its maximum emission wavelength was found at 545 nm. Optimal conditions for the formation of the levofloxacin–Tb(III) complexes were studied. Levofloxacin was detected by measuring the FL intensity, which increases linearly with the concentration of LVX in the range 6.2 × 10−10–2.6 × 10−8 mol/L. Recovery of the target analytes was > 96% with good quality parameters: linearity (r2 > 0.996), limit of detection (LOD) and limit of quantification (LOQ) values 2.1 × 10−10 mol/L and 7.2 × 10−10 mol/L, and run‐to‐run and day‐to‐day precisions with relative standard deviations (RSDs) around 3%. Thus, the proposed method can be successfully applied to the routine determination of levofloxacin in pharmaceutical preparations. Copyright © 2011 John Wiley & Sons, Ltd.  相似文献   

8.
Approximately 80 per cent of tyrosine hydroxylase activity in bovine mandibular nerve and rabbit sciatic nerve was soluble, and the rest of the activity was particle-bound. The soluble enzyme in bovine mandibular nerve was isolated by ammonium sulphate fractionation (25–35 per cent saturation). The enzyme had a pH optimum at 5·9 in Tris-acetate buffer, and at 6·5 in Tris-HCl or phosphate buffer. The enzyme required a tetrahydropteridine cofactor. Km values toward various tetrahydropteridines such as l -erythro-tetrahydrobiopterin (a probable natural cofactor), 2-amino-4-hydroxy-6-methyltetrahydropteridine, and 2-amino-4-hydroxy-6,7-dimethyltetrahydropteridine were 2 × 10−5m , 5 × 10−5m and 4 × 10−4m , respectively. The Km value for tyrosine at 1 × 10−3m -2-amino-4-hydroxy-6-methyltetrahydropteridine as a cofactor was 5 × 10−5m . The enzyme activity was markedly stimulated with Fe2+ or catalase, but Fe2+ gave higher activity. The activity was inhibited with α, α′-dipyridyl, l -α-methyl-p-tyrosine, and various catecholamines. Among catecholamines, dopamine was the most potent inhibitor. l -5-Hydroxytryptophan was an inhibitor as potent as dopamine. Neither d -5-hydroxytryptophan nor 5-hydroxytryptamine inhibited the enzyme. The inhibition by l -5-hydroxytryptophan was partially competitive with tetrahydrobiopterin at concentrations higher than 9 × 10−5m , and partially uncompetitive at concentrations lower than 9 × 10−5m . The addition of heparin or lysolecithin did not affect enzyme activity with tetrahydrobiopterin as cofactor.  相似文献   

9.
《Insect Biochemistry》1990,20(6):557-566
[125I]α-Bungarotoxinisusedasaprobetostudythenicotinic-cholinergicreceptorinmembrane preparations of the cockroach brain. Binding is restricted mainly to particulate fractions of brain homogenates, is time dependent and is saturable above 2 nM with very low non-specific binding. Scatchard analysis indicates that binding is associated with a single affinity site (Kd = 1.09 nM) having a Bmax of 8926 fmol/mg protein which is the highest concentration of binding sites yet reported in insects. Association kinetics are best fit by a mono-exponential model with a kobs = 4.37 × 10−3s−1. Dissociation is best described by a bi-exponential model giving dissociation constants of 1.18 × 10−5 and 9.94 × 10−5s−1. The Kds calculated from kinetic data are 0.029 and 0.25 nM suggesting the possibility of heterogeneous binding sites not detected by saturation studies. Displacement studies indicate that binding follows a nicotinic pharmacology and demonstrate the high affinity of methyllycaconitine and the anthelmintics, morantel and pyrantel. Displacement by neuronal bungarotoxin shows the presence of two distinct binding sites not differentiated by α-bungarotoxin. Autoradiographic studies show α-bungarotoxin to be binding to neuropile regions of the brain, to be displaced from these regions by agents effective in binding studies and demonstrate that the neuronal bungarotoxin binding sites can be regionally localized.  相似文献   

10.
Based on the strong electrochemiluminescence (ECL) reaction between thiamazole and tris(2,2′‐bipyridine)ruthenium(II) (Ru(bpy)32+), a sensitive, simple and rapid flow injection analysis method for the determination of thiamazole was developed. When a Pt working electrode was maintained at a potential of +1.50 V (vs Ag/AgCl) in pH 12.0 H3PO4–NaOH solution containing thiamazole and Ru(bpy)32+ at a flow rate of 1.0 mL/min, a linear range of 2.0 × 10−7–1.0 × 10−4 mol/L with a detection limit of 5.0 × 10−8 mol/L was obtained for the detection of thiamazole. The method showed good reproducibility with a relative standard deviation (RSD) of 0.75%. The method has been successfully applied to the determination of thiamazole in spiked animal feeds. In addition, a co‐reactant ECL mechanism was proposed for the thiamazole–Ru(bpy)32+ system. Copyright © 2014 John Wiley & Sons, Ltd.  相似文献   

11.
Sensitive detection of doxorubicin (DOX) is critical for clinical theranostics. A novel ratiometric fluorescence strategy based on the inner filter effect (IFE) has been established for the sensitive detection of DOX by designing a ratiometric fluorescence probe. In the presence of DOX, the fluorescence intensity of copper nanoclusters (CuNCs) at 485 nm decreases, and the fluorescence intensity of carbon dots at 560 nm increases. Therefore, DOX can be quantitatively detected by measuring the ratio of the fluorescence intensities at 560 and 485 nm (F560/F485). The F560/F485 ratio exhibits a linear correlation with the DOX concentration in the range from 1.0 × 10−8 M to 1.0 × 10−4 M with the detection limit of 3.7 nM. Furthermore, this method was also successfully applied to the analysis of DOX in human plasma samples, affording an effective platform for drug safety management.  相似文献   

12.
In this study, 5,10,15,20-(4-sulphonatophenyl)porphyrin (TPPS4) was selected as a fluorescent probe due to its excellent characteristics including high quantum yield, good water solubility, and exceptional biocompatibility. With an excitation wavelength set at 515 nm, the optimal fluorescence emission wavelength for TPPS4 was measured at 642 nm. At this moment, the fluorescence signal of TPPS4 pink solution was in the ‘ON’ state. The fluorescence intensity of TPPS4 was quenched when ascorbic acid (AA) was introduced, which was due to the electron transfer quenching effect between AA and TPPS4. The colour of the corresponding solution changed from pink to green, and the fluorescence signal was in the ‘OFF’ state. When HPO42− was further introduced into the TPPS4–AA system, the quenched fluorescence intensity of TPPS4 was recovered due to the unique interaction between HPO42− and AA. At this time, the colour of the corresponding solution changed from green to red, and the fluorescence signal was in the ‘ON’ state. Therefore, an ‘ON–OFF–ON’ signal-switchable fluorescent probe was constructed based on TPPS4 to detect HPO42−. The results showed that the linear range of HPO42− was 4.0 × 10−9 to 1.7 × 10−6 M, and the detection limit was 1.3 × 10−9 M (S/N = 3). The sensing system exhibited high accuracy and sensitivity, and it could be used successfully to detect HPO42− in real samples.  相似文献   

13.
Du  Bobo  Yang  Yuan  Zhang  Yang  Yang  Dexing 《Plasmonics (Norwell, Mass.)》2019,14(2):457-463

In this article, a surface plasmon resonance (SPR) biosensor based on D-typed optical fiber coated by Al2O3/Ag/Al2O3 film is investigated numerically. Resonance in near infrared with an optimized architecture is achieved. Refractive index sensitivity of 6558 nm/RIU (refractive index unit) and detection limit of 1.5 × 10−6 RIU, corresponding to 0.4357 nm/μM and detection limit of 23 nM in BSA (bovine serum albumin) concentration sensing, are obtained. The analysis of the performance of the sensor in gaseous sensing indicates that this proposed SPR sensor is much suitable for label-free biosensing in aqueous media.

  相似文献   

14.
《Luminescence》2018,33(3):625-629
A simple, sensitive and efficient fluorescence method has been established for the quantitative analysis of bilirubin. The fluorometric determination method was based on the kinetic quenching of ruthenium(II) fluorescence. The quenching effect may be due to the complexation reaction of bilirubin with ruthenium(II). Therefore, the effects of ruthenium concentrations and different surfactants have been studied. Under the optimized experimental parameters, the fluorescence intensity decreased proportionally with the bilirubin concentration and linearity was established in the range of 3.3 × 10−7 to 3.0 × 10−4 M bilirubin. The detection limit calculated from the calibration graph was found to be 5.2 × 10−8 M. The relative standard deviation (RSD) of 10 consecutive measurements of 8.0 × 10−6 M bilirubin was 3.0%, while the recoveries of bilirubin in both human serum and urine samples were obtained in the range 94.0–99.5%. The interference study shows that the developed fluorescence based technique is fast, easy to carry out and shows negligible interference. The developed technique was successfully applied for the analysis of bilirubin in human urine and serum samples. All the experimental results and quality parameters confirmed the sensitivity and reproducibility of the proposed technique for bilirubin determination in human urine and serum samples .  相似文献   

15.
Abstract— —The influx of glutamic acid in frog sciatic nerve has been studied by monitoring the disappearance of 14C labelled compound from the bathing medium. After 5hr of incubation in 10 −6m non-labelled l -glutamic acid and 0·01, μc/ml labelled isotope, the intracellular concentration of labelled glutamic acid is about 15 times the concentration in the bathing medium; however, there appears to be a net loss of non-labelled compound with incubation. Uptake of L,-glutamic acid is accompanied by conversion of significant amounts of labelled E-glutamic acid to carbon dioxide and glutamine; small amounts of γ-aminobutyric acid and aspartic acid are also formed. The rate of disappearance of labelled l -glutamic acid decreases with increasing concentration of non-labelled isotope in the bathing medium. Construction of a Lineweaver-Burk plot from initial velocities of influx yields an average Vm of 4·02 × 10−9 moles/g/min and an average Km. of 3·23 × 10 −5 moles/l. The influx of glutamic acid is highly specific with regard to molecular structure; of the compounds tested, only l -glutamine, l -glutamic acid, GABA, l -lysine, and l -aspartic acid are taken up, and only l -aspartic acid will compete with l -glutamic acid for uptake.  相似文献   

16.
An economical and green approach to the synthesis of naphthyl derivative for detection of l -carnitine (3-hydroxy-4-N-trimethyl-aminobutyrate) is practically important. We developed a naphthyl derivative as a probe showing ‘turn-on’ response towards l -carnitine selectively at pH 7.2 through ICT mechanism with a good limit of detection (LOD) of 0.126 μM. Using Job's plot for determining the binding stoichiometry, it was found that probe could form a more stable complex (1:1) with carnitine. The binding constant (K) between probe and carnitine was calculated as 8 × 107 M−1 using the Benesi–Hildebrand plot. The binding interaction of the probe with l -carnitine was confirmed by nuclear magnetic resonance titrations, Fourier-transform infrared spectroscopy, photo physical studies and density functional theory calculations. Meanwhile, the probe can be used to quantitatively detect carnitine in food samples.  相似文献   

17.
The effects of prostaglandin F (PGF) on propulsive activity in segments of isolated colon and on isolated strips of guinea-pig colon were investigated.Using experimental conditions under which spontaneous propulsive activity was negligible, PGF (5×10−8×1×10−6M), added to the bathing medium, increased propulsive activity in a concentration dependent manner. This increase of propulsive activity was abolished in the presence of atropine or tetrodotoxin (1×10−7g/ml).The contractions produced by PGF(5×10−7 − 1×10−5M) in isolated longitudinal and circular smooth muscle strips of guinea-pig colon were unaffected in the presence of atropine or tetrodotoxin (1×10−7g/ml).From these results it is concluded that under the conditions employed in this study propulsive activity stimulated by PGF may depend on the contractions of both muscle layers and stimulation of the peristalic reflex.  相似文献   

18.
A photochemical flow-injection methodology for the fluorimetric determination of fenvalerate, a nonfluorescent pyrethroid pesticide, is proposed. The sample was irradiated by ultraviolet light inside a reaction coil in a nonstop mode and the photodegradation products were monitored by spectrofluorimetry at λexc=277 nm, λem=329 nm. The exploitation of a micellar medium resulted in a pronounced fluorescence enhancement with a concomitant tenfold sensitivity improvement. Several surfactants were evaluated, and best results were obtained with sodium dodecylsulphate (SDS). The calibration plot was linear within 2.0×10−6 and 1.0×10−5 mol l−1 fenvalerate. The proposed system is very stable, handles 30 samples h−1 and requires only 60 μmol SDS per determination. The detection limit is 5×10−7 mol l−1. The method was evaluated in the determination of fenvalerate in tap water.  相似文献   

19.
Proteomic approaches have unique advantages in the identification of biological pathways that influence physical frailty, a multifactorial geriatric syndrome predictive of adverse health outcomes in older adults. To date, proteomic studies of frailty are scarce, and few evaluated prefrailty as a separate state or examined predictors of incident frailty. Using plasma proteins measured by 4955 SOMAmers in the Atherosclerosis Risk in Community study, we identified 134 and 179 proteins cross-sectionally associated with prefrailty and frailty, respectively, after Bonferroni correction (p < 1 × 10−5) among 3838 older adults aged ≥65 years, adjusting for demographic and physiologic factors and chronic diseases. Among them, 23 (17%) and 82 (46%) were replicated in the Cardiovascular Health Study using the same models (FDR p < 0.05). Notably, higher odds of prefrailty and frailty were observed with higher levels of growth differentiation factor 15 (GDF15; pprefrailty = 1 × 10−15, pfrailty = 2 × 10−19), transgelin (TAGLN; pprefrailty = 2 × 10−12, pfrailty = 6 × 10−22), and insulin-like growth factor-binding protein 2 (IGFBP2; pprefrailty = 5 × 10−15, pfrailty = 1 × 10−15) and with a lower level of growth hormone receptor (GHR, pprefrailty = 3 × 10−16, pfrailty = 2 × 10−18). Longitudinally, we identified 4 proteins associated with incident frailty (p < 1 × 10−5). Higher levels of triggering receptor expressed on myeloid cells 1 (TREM1), TAGLN, and heart and adipocyte fatty-acid binding proteins predicted incident frailty. Differentially regulated proteins were enriched in pathways and upstream regulators related to lipid metabolism, angiogenesis, inflammation, and cell senescence. Our findings provide a set of plasma proteins and biological mechanisms that were dysregulated in both the prodromal and the clinical stage of frailty, offering new insights into frailty etiology and targets for intervention.  相似文献   

20.
Two Schiff base probes ( S1 and S2 ) were prepared and synthesized by incorporating thienopyrimidine into salicylaldehyde or 3-ethoxysalicylaldehyde individually, with the aim of detecting Ga3+ and Pd2+ sequentially. Upon chelation with Ga3+, S1 and S2 exhibited fluorescence enhancement in DMSO/H2O buffer. Both S1 –Ga3+ and S2 –Ga3+ were quenched by Pd2+. The limit of detection for S1 in response to Ga3+ and Pd2+ was 2.86 × 10−7 and 4.4 × 10−9 M, respectively. For S2 , the limit of detection for Ga3+ and Pd2+ was 4.15 × 10−8 and 3.0 × 10−9 M, respectively. Furthermore, the complexation ratios of both S1 and S2 with Ga3+ and Pd2+ were determined to be 1:2 through Job's plots, ESI-MS analysis, and theoretical calculations. Two molecular logic gates were constructed, leveraging the response behaviors of S1 and S2 . Moreover, the potential utility of S1 and S2 for monitoring Ga3+ and Pd2+ in domestic water was verified.  相似文献   

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