Synthesis of CuInS2 Quantum Dots Modified with Alanine for Sensitive and Selective Detection of Pb2+ and Cysteine.

In this study, a novel fluorescent probe based on CuInS2 quantum dots modified with alanine (Ala-CuInS2 QDs) was developed for the detection of lead ions and cysteine (Pb2+ and Cys). Ala-CuInS2 QDs were synthesized through a one-step hydrothermal method exhibiting uniform size, good stability and water solubility. The QDs were then utilized as an "on-off-on" fluorescence sensor to detect Pb2+ and Cys in the ranges of 0-20 µM and 0-55 µM respectively, with detection limits of 0.29 µM and 0.66 µM. The mechanism of fluorescence quenching and recovery processes was also explored. Furthermore, Ala-CuInS2 QDs have been successfully applied to detect Pb2+ in tap and river water and detect cysteine in serum.

Multifunctional N, Fe-doped carbon dots with peroxidase-like activity for the determination of H2O2 and ascorbic acid and cell protection against oxidation.

Multifunctional N, Fe-doped carbon dots (N, Fe-CDs) were synthesized by the one-step hydrothermal method using ferric ammonium citrate and dicyandiamide as raw materials. The N, Fe-CDs exhibited peroxidase-like (POD) activity by catalyzing the oxidization of 3,3',5,5'-tetramethylbenzidine (TMB) to the green oxidation state ox-TMB in the presence of hydrogen peroxide (H2O2). Subsequently, based on the POD activity of N, Fe-CDs, an efficient and sensitive colorimetric method for the detection of H2O2 and ascorbic acid (AA) was established with a limit of detection of 0.40 µM and 2.05 µM. The proposed detection method has been successfully applied to detect AA in fruit juice, vitamin C tablets, and human serum samples and has exhibited excellent application prospects in biotechnology and food fields. Furthermore, N, Fe-CDs also showed a protective effect on the cell damage caused by H2O2 and could be used as an antioxidant agent.

N-doped carbon dots as the multifunctional fluorescent probe for mercury ion, glutathione and pH detection.

Recently, carbon dots (CDs) have exhibited promising applications in the fluorescence detection of various ions and biomolecules. In this work, one kind of nitrogen-doped CDs (N-CDs) with high fluorescence intensity was synthesized, characterized by transmission electron microscopy, x-ray diffraction, x-ray photoelectron spectroscopy, Fourier-transform infrared, UV-vis absorption spectra, and fluorescence spectra. The results show that the spherical and uniform N-CDs (quantum yield: 60.2%) have remarkable fluorescence properties and photostability, which makes N-CDs can be utilized as an 'on-off-on' sensor for Hg2+and glutathione (GSH). In addition, the pH-sensitive behavior of N-CDs makes it also applicable to H+detection under acid conditions (pKa = 3.53). The linear range of the 'turn-off' sensor detecting Hg2+was 0.014-50µM, with a 0.014µM limit of detection (LOD). GSH was detected by the fluorescence 'turn-on' method with a linear range of 0.125-60µM and a LOD of 0.125µM. The outstanding performance of N-CDs makes it potential applications in ecological pollution and biomolecule visualization monitoring.

The on-off-on Fluorescence Sensor of Hollow Carbon Dots for Detecting Hg2+ and Ascorbic Acid.

Carbon dots (CDs) have excellent fluorescence properties and can be used in many research fields. In this paper, carbon dots were prepared by microwave-assisted pyrolysis of citric acid and urea, characterized by transmission electron microscope (TEM), X-ray diffractometer (XRD), 13C-NMR spectrum, zeta potential, Fourier transform infrared spectroscopy (FT-IR), ultraviolet-visible (UV-vis) absorption and fluorescence spectra, and detected the Hg2+ and ascorbic acid (AA) sequentially. It showed that carbon dots were hollow, spherical particles and less than 10 nm, photoluminescence quantum yield of carbon dots was about 15%. The CDs were selective and sensitive to Hg2+ and AA based on the "on-off-on" fluorescence behavior. The detection limits of CDs for Hg2+ and AA were 0.138 µM and 0.212 µM, respectively. Fluorescence response mechanism of CDs was also discussed.

In-cell destabilization of a homodimeric protein complex detected by DEER spectroscopy.

The complexity of the cellular medium can affect proteins' properties, and, therefore, in-cell characterization of proteins is essential. We explored the stability and conformation of the first baculoviral IAP repeat (BIR) domain of X chromosome-linked inhibitor of apoptosis (XIAP), BIR1, as a model for a homodimer protein in human HeLa cells. We employed double electron-electron resonance (DEER) spectroscopy and labeling with redox stable and rigid Gd3+ spin labels at three representative protein residues, C12 (flexible region), E22C, and N28C (part of helical residues 26 to 31) in the N-terminal region. In contrast to predictions by excluded-volume crowding theory, the dimer-monomer dissociation constant KD was markedly higher in cells than in solution and dilute cell lysate. As expected, this increase was partially recapitulated under conditions of high salt concentrations, given that conserved salt bridges at the dimer interface are critically required for association. Unexpectedly, however, also the addition of the crowding agent Ficoll destabilized the dimer while the addition of bovine serum albumin (BSA) and lysozyme, often used to represent interaction with charged macromolecules, had no effect. Our results highlight the potential of DEER for in-cell study of proteins as well as the complexities of the effects of the cellular milieu on protein structures and stability.

Effect of Steam on Carbonation of CaO in Ca-Looping.

Ca-looping is an effective way to capture CO2 from coal-fired power plants. However, there are still issues that require further study. One of these issues is the effect of steam on the Ca-looping process. In this paper, a self-madethermogravimetric analyzer that can achieve rapid heating and cooling is used to measure the change of sample weight under constant temperature conditions. The parameters of the Ca-looping are studied in detail, including the addition of water vapor alone in the calcination or carbonation stage and the calcination/carbonation reaction temperatures for both calcination and carbonation stages with water vapor. Steam has a positive overall effect on CO2 capture in the Ca-looping process. When steam is present in both calcination and carbonation processes, it increases the decomposition rate of CaCO3 and enhances the subsequent carbonation conversion of CaO. However, when steam was present only in the calcination process, there was lower CaO carbonation conversion in the following carbonation process. In contrast, when steam was present in the carbonation stage, CO2 capture was improved. Sample characterizations after the reaction showed that although water vapor had a negative effect on the pore structure, adding water vapor increased the diffusion coefficient of CO2 and the carbonation conversion rate of CaO.

New Process Combining Fe-Based Chemical Looping and Biomass Pyrolysis for Cogeneration of Hydrogen, Biochar, Bio-Oil and Electricity with In-Suit CO2 Separation.

Fe-based chemical looping gasification is a clean biomass technology, which has the advantage of reducing CO2 emissions and the potential of self-sustaining operation without supplemental heating. A novel process combining Fe-based chemical looping and biomass pyrolysis was proposed and simulated using Aspen Plus. The biomass was first subjected to pyrolysis to coproduce biochar, bio-oil and pyrolysis gas; the pyrolysis gas was subjected to an Fe looping process to obtain high-purity hydrogen and carbon dioxide. The influences of the pyrolysis reactor operating temperature and fuel reactor operation temperature, and the steam reactor and air reactor on the process performance are researched. The results showed that, under the operating condition of the established process, 23.07 kg/h of bio-oil, 24.18 kg/h of biochar, 3.35 kg/h of hydrogen and a net electricity of 3 kW can be generated from 100 kg/h of rice straw, and the outlet CO2 concentration of the fuel reactor was as high as 80%. Moreover, the whole exergy efficiency and total exergy loss of the proposed process was 58.98% and 221 kW, respectively. Additionally, compared to biomass direct chemical looping hydrogen generation technology, the new process in this paper, using biomass pyrolysis gas as a reactant in the chemical looping hydrogen generation process, can enhance the efficiency of hydrogen generation.

Colorimetric and ultra-sensitive fluorescence resonance energy transfer determination of H2O2 and glucose by multi-functional Au nanoclusters.

Ultra-sensitive colorimetric determination of H2O2 is accomplished based on the intrinsic peroxidase-like activity of Au nanoclusters (AuNCs) stabilized by glutathione (GSH). The color change of 3,3,5,5-tetramethylbenzidine (TMB) catalyzed by AuNCs offers an indirect method to measure glucose. This sensing platform makes use of a dual optical signal change, including the color change in an aqueous solution under visible light illumination and an ultra-sensitive fluorescent assay arising from efficient fluorescence resonance energy transfer (FRET) between the AuNCs and oxidized TMB. The detection limits of H2O2 and glucose are 4.9 × 10(-13) M and 1.0 × 10(-11) M, respectively. In addition, enhanced fluorescence is observed from the AuNCs due to the use of ethanol which produces clear changes in the quantum yield and lifetime of the AuNCs. The quantum yield of AuNCs is enhanced from ∼12.5% as an isolated fluorophore to 38.9% in an AuNCs-ethanol complex. The enhanced fluorescence lowers the detection limits of H2O2 and glucose by 2 orders of magnitude compared to those attained from the original AuNCs.

Ultrasensitive determination of copper in complex biological media based on modulation of plasmonic properties of gold nanorods.

Accurate determination of copper in complex biological media such as cells is quite difficult, and an analytical strategy based on copper-modulated formation of core-shell gold nanorods is described. Selective and label-free sensing can be achieved by measuring the change in the localized surface plasmon resonance absorption. The technique can determine trace amounts of copper in human serum, urine, and red blood cells without or with minimal sample pretreatment. The Cu detection limits are 20.67 µM in human serum, 0.193 µM in human urine, and 3.09 × 10(-16) g in a single cell. The advantages of the technique are the high selectivity, simple or no sample pretreatment, and label free. Boasting a practical detection limit down to 2 fM, only 10(3) red blood cells are needed to conduct the analysis and the technique may be extended to the detection of trace amounts of copper in a single cell.

Multiplex plasmonic sensor for detection of different metal ions based on a single type of gold nanorod.

In this paper, a label-free multiplex plasmonic sensor has been developed to selectively determine different metal ions including Fe(3+), Hg(2+), Cu(2+), and Ag(+) ions based on a single type of gold nanorod (GNR). Under proper conditions, these metal ions can react with GNRs, resulting in changes of nanostructure and composition. The determination of Fe(3+), Hg(2+), Cu(2+), and Ag(+) ions is therefore readily implemented due to changes of longitudinal plasmon wavelength (LPW) of nanorods. Moreover, the GNR-based assay can not only determine all four kinds of metal ions successively but also can detect which of any one or several kinds of metal ions. This assay is sensitive to detect Fe(3+), Hg(2+), Cu(2+), and Ag(+) as low as 10(-6), 10(-8), 10(-10), and 10(-8) M, respectively. Importantly, the special nanostructure and composition of the nanorods are induced by these metal ions, which allow this sensor to maintain high selectivity to determine these metal ions. This nanosensor abrogates the need for complicated chemosensors or sophisticated equipment, providing a simple and highly selective detection platform.

Nano-Resveratrol Liposome: Physicochemical Stability, In Vitro Release, and Cytotoxicity.

Nano-resveratrol liposome (RES-LIP) was prepared by the thin film rotary-evaporated method combined with ultrasonication and characterized by transmission electron microscopy (TEM), zeta potential, dynamic light scattering (DLS), and Fourier-transform infrared (FT-IR). The physicochemical stability, in vitro release, antioxidant activity, and cytotoxicity of RES-LIP were studied. Data showed that RES-LIP was a spherical vesicle with a diameter of less than 100 nm, the zeta potential was - 60 mV and the encapsulation efficiency was 86.78%. The physicochemical stability of RES-LIP was determined by Ea, ΔG, ΔH, and ΔS, which suggested that the process of RES-LIP degradation was spontaneous and endothermic. The in vitro release of RES-LIP was pH-dependent, belonged to the Weibull model, and was non-Fick diffusion. The antioxidant activity of RES-LIP was stronger than free resveratrol. The MTT assay and flow cytometry results suggested that resveratrol decreased cytotoxicity after being encapsulated by liposome. The prepared RES-LIP had high encapsulation efficiency, was sustained-release, had low cytotoxicity, was pH-targeted, and had potential usage in food and medicine fields.

High Drug Capacity of Nano-Levodopa-Liposomes: Preparation, In Vitro Release and Brain-Targeted Research.

In this work, nano-levodopa-liposomes (L-dopa-Lip) suspension was prepared by rotary-evaporated film-ultrasonic method, and freeze-drying powders of L-dopa-Lip were also obtained to improve the stability. The products were characterized by TEM, DLS, and TG-DSC, and the phase-transition temperature (Tm) and encapsulation efficiency were calculated. The brain-targeting and in vitro release of the drug was also studied. The results showed that L-dopa-Lip were well-formed spherical vesicles, and the sizes were about 100 nm, and the encapsulation efficiency was higher than 90%. The drug release temperature of L-dopa-Lip was 68 °C, and the in vitro release property and mathematical model were also studied. The brain targeting of L-dopa-Lip in vivo was explored by injecting the gold nanoparticles (AuNPs) labeled L-dopa-Lip (AuNPs-L-dopa-Lip) through the tail vein. ICP-MS and TEM showed that L-dopa-Lip had brain targeting, suggesting the potential treatment of L-dopa-Lip on brain dysfunction. The results of this work might be helpful for designing drug-loaded liposomes for the treatment of central nervous system (CNS) diseases and monitoring their distributions in vivo.

Third BIR domain of XIAP binds to both Cu(II) and Cu(I) in multiple sites and with diverse affinities characterized at atomic resolution.

The X-chromosome linked inhibitor of apoptosis, XIAP, is mainly known as the inhibitor of caspases by direct interaction with caspases with its baculoviral IAP repeat (BIR) domains. XIAP has three BIR domains and each BIR domain contains a zinc binding site, normally known as zinc finger motif. Recent studies showed that XIAP is involved in copper homeostasis in cells and the BIR domains bind copper ion. However, structural details of the second and third BIR domain, BIR2 and BIR3, in XIAP, with copper as well as the binding modes are not known. In the present work we characterize the structural properties of BIR3 in solution by high resolution NMR and other biophysical techniques. The interaction of BIR3 with copper both in vitro and in cell lysates was analyzed. Our results show that BIR3 is able to form stable complexes both with Cu(II) and Cu(I), whereas zinc binding site is not affected and zinc retains tightly bound in the zinc finger during these interactions. Surprisingly, BIR3 has multiple binding sites for Cu(II) and Cu(I) but with varied binding affinities. In addition, the solvent exposed Cys351 is readily oxidized by Cu(II) resulting an intermolecular disulfide bond either between two BIR3 molecules or a mixed disulfide bond with glutathione in cell lysates.

Solution structure and interaction with copper in vitro and in living cells of the first BIR domain of XIAP.

The X-chromosome linked inhibitor of apoptosis (XIAP) is a multidomain metalloprotein involved in caspase inhibition and in copper homeostasis. It contains three zinc-binding baculoviral IAP repeats (BIR) domains, which are responsible for caspase interaction. Recently, it has been suggested that the BIR domains can bind copper, however high resolution data on such interaction is missing. Here we characterize by NMR the structural properties of BIR1 in solution, and the effects of its interaction with copper both in vitro and in physiological environments. BIR1 is dimeric in solution, consistent with the X-ray structure. Cysteine 12, located in the unfolded N-terminal region, has a remarkably low redox potential, and is prone to oxidation even in reducing physiological environments. Interaction of BIR1 with copper(II) results in the oxidation of cysteine 12, with the formation of either an intermolecular disulfide bond between two BIR1 molecules or a mixed disulfide bond with glutathione, whereas the zinc binding site is not affected by the interaction.

Alternate release of different target species based on the same gold nanorods and monitored by cell imaging.

In this study, a strategy for load and release of different kinds of molecules on the same gold nanorods (GNRs) was developed. An anticancer drug, doxorubicin hydrochloride (DOX), was firstly chemically conjugated on the GNRs. To efficiently load another type of target molecules DNA on the same GNRs, a polyelectrolyte Poly (ethylene imine) (PEI) was adsorbed on the GNR@DOX to form GNR@DOX@PEI. Then, the positive charge GNR@DOX@PEI allows the GNR conjugates to interact with negative charged DNA by an electrostatic interaction, enabling their full conjugation. A platform to load two kinds of target molecules conjugated on the same GNRs was fabricated. On the other hand, selective and sequential release of the different target species may be triggered by chemical reaction and near infrared (NIR) laser. The release of DOX was achieved by Na2S2O3 reacting with GNRs and the discharge of DNA conjugated on the GNR@DOX@PEI was accomplished by local-heating using NIR laser triggered release. Furthermore, the selective and alternate release of different target species from the GNRs inside MCF-7 cells was monitored by fluorescent imaging, providing a potential synergistic cancer treatment.

Carbon dots as a fluorescent probe for label-free detection of physiological potassium level in human serum and red blood cells.

A unique photoluminescence carbon dots (CDs) with larger size were prepared by microwave-assisted method. Complex functional groups on the surface of the CDs facilitate the nanoparticles to form affinity with some metal ions. Taking advantage of the effective fluorescence quenching effect of K(+), a highly sensitive CD-based fluorescence analytical system for label-free detection of K(+) with limit of detection (LOD) 1.0×10(-12) M was established. The concentrations of potassium ion in biological samples such as human serum are usually found at millimolar levels or even higher. The proposed method begins with a substantial dilution of the sample to place the K(+) concentration in the dynamic range for quantification, which covers 3 orders of magnitude. This offers some advantages: the detection of K(+) only needs very small quantities of biological samples, and the dilution of samples such as serum may effectively eliminate the potential interferences that often originate from the background matrix. The determined potassium levels were satisfactory and closely comparable with the results given by the hospital, indicating that this fluorescent probe is applicable to detection of physiological potassium level with high accuracy. Compared with other relative biosensors requiring modified design, bio-molecular modification or/and sophisticated instruments, this CD-based sensor is very simple, cost-effective and easy detection, suggesting great potential applications for successively monitoring physiological potassium level and the change in biological system.

Multiplex sensor for detection of different metal ions based on on-off of fluorescent gold nanoclusters.

In this study, a multiplex fluorescence sensor for successive detection of Fe(3+), Cu(2+) and Hg(2+) ions based on "on-off" of fluorescence of a single type of gold nanoclusters (Au NCs) is described. Any of the Fe(3+), Cu(2+) and Hg(2+) ions can cause quenching fluorescence of Au NCs, which established a sensitive sensor for detection of these ions respectively. With the introduction of ethylene diamine tetraacetic acid (EDTA) to the system of Au NCs and metal ions, a restoration of fluorescence may be found with the exception of Hg(2+). A highly selective detection of Hg(2+) ion is, thus, achieved by masking Fe(3+) and Cu(2+). On the other hand, the masking of Fe(3+) and Cu(2+) leads to the enhancement of fluorescence of Au NCs, which in turn provides an approach for successive determination of Fe(3+) and Cu(2+) based on "on-off" of fluorescence of Au NCs. Moreover, this assay was applied to the successful detection of Fe(3+), Cu(2+) and Hg(2+) in fish, a good linear relationship was found between these metal ions and the degree of quenched fluorescent intensity. The dynamic ranges of Hg(2+), Fe(3+) and Cu(2+) were 1.96×10(-10)-1.01×10(-9), 1.28×10(-7)-1.27×10(-6) and 1.2×10(-7)-1.2×10(-6) M with high sensitivity (the limit of detection of Fe(3+) 2.0×10(-8) M, Cu(2+) 1.9×10(-8) M and Hg(2+) 2×10(-10) M). These results indicate that the assay is suitable for sensitive detection of these metal ions even under the coexistence, which can not only determine all three kinds of metal ions successively but also of detecting any or several kinds of metal ions.

Enhanced fluorescence of chitosan based on size change of micelles and application to directly selective detecting Fe³âº in human serum.

In this paper, we have developed an approach to significantly enhance fluorescence of chitosan by simply heated the inherently low fluorescent chitosan aqueous solution. Enhanced blue fluorescence of chitosan solution was observed as originated from the formation of small size of chitosan micelle after long time heated. The fluorescence of chitosan micelles was quenched and recovered when Fe³âº ions were combined and released from chitosan micelles. Therefore, chitosan without modification of functional groups can recognize Fe³âº with very high selectivity. As a result, a new fluorescence sensor for sensitively detecting Fe³âº ion based on the change of chitosan micelles sizes was subsequently fabricated. This enhanced fluorescence enables the chitosan sensor to be sensitive to low concentrations of Fe³âº, and it is linear responsive in the range of 1.96×10⁻8 to 2.00×10⁻5 M. Importantly, this novel sensor may be applied in human serum for direct detection of Fe³âº ion without sample pretreatment. Analysis of 5 samples of human serum shows that the average concentration of Fe³âº is 26.95 µM, which is consistent with the results determined by other methods. Moreover, the advantage of chitosan-based assay is that Fe³âº rather than Fe²âº in human serum can be directly measured, avoiding costly, time-consuming and complex process.