A ratiometric fluorescent assay for the detection and bioimaging of alkaline phosphatase based on near infrared Ag2S quantum dots and calcein.

Herein, a ratiometric fluorescent method was developed for alkaline phosphatase (ALP) detection based on near-infrared (NIR) Ag2S quantum dots (QDs) and calcein through the competitive approach. The system based on Ag2S QDs and calcein shows green (maximum emission at 512 nm from calcein) and near infrared (NIR) fluorescence (maximum 798 nm from Ag2S QDs) under the same excitation wavelength (468 nm). In the presence of Ce3+, the fluorescence intensity of calcein is decreased due to static quenching, while the fluorescence intensity of Ag2S QDs is enhanced through aggregation induced emission (AIE). The p-nitrophenyl phosphate is hydrolyzed by ALP, and the yield phosphate ions bind with Ce3+ with higher affinity than these of Ag2S QDs and calcein. Therefore, the green fluorescence from calcein is recovered while NIR fluorescence from Ag2S QDs is decreased. On the basis of these findings, a ratiometric fluorescence assay was developed for the measurement of ALP activity. The ratio of fluorescence intensity at 512 and 798 nm (F512/F798) was well associated with the ALP concentration ranging from 2 to 100 mU/mL with the detection limit of 1.28 mU/mL. The method was successfully applied for detecting ALP in human serum with an acceptable recovery and bioimaging intracellular ALP with good performance. In addition, the approach was also employed for the screening ALP inhibitor.

Label-free fluorometric assay for cytochrome c in apoptotic cells based on near infrared Ag2S quantum dots.

As an important biomarker, cytochrome c (Cyt c) plays a crucial role in mitochondrial electron transport chain and cell apoptosis. Herein, a label-free near-infrared (NIR) Ag2S quantum dots (QDs)-based fluorescent strategy was constructed for the sensitive and selective detection of Cyt c. In this system, Cyt c was hydrolyzed by trypsin, and the resulting heme-peptide fragment exhibited peroxidase-like activity for catalytic decomposition of H2O2 into hydroxyl radical (·OH). The presence of caffeic acid in this system resulted into the formation of caffeic acid-quinone due to the strong oxidizing ability of ·OH. The production of caffeic acid-quinone led to the fluorescence quenching of Ag2S QDs through electron transfer mechanism. Based on the cascade reaction, we successfully developed a label-free approach to detect Cyt c using Ag2S QDs as nanoprobes. Under the optimized conditions, the fluorescence intensity of Ag2S QDs was linearly relative to the concentration of Cyt c over the range from 2.0 to 150 nM with a detection limit of 1.7 nM. In addition, this strategy was successfully applied for quantitative detection of Cyt c in cell lysates of H2O2 or etoposide (anticancer drug)-induced apoptotic cells, providing great potential application for cell-based oxidation pressure determination and screening of anticancer drugs.

Near-Infrared Fluorescent Ag2S Nanodot-Based Signal Amplification for Efficient Detection of Circulating Tumor Cells.

The level of circulating tumor cells (CTCs) plays a critical role in tumor metastasis and personalized therapy, but it is challenging for highly efficient capture and detection of CTCs because of the extremely low concentration in peripheral blood. Herein, we report near-infrared fluorescent Ag2S nanodot-based signal amplification combing with immune-magnetic spheres (IMNs) for highly efficient magnetic capture and ultrasensitive fluorescence labeling of CTCs. The near-infrared fluorescent Ag2S nanoprobe has been successfully constructed through hybridization chain reactions using aptamer-modified Ag2S nanodots, which can extremely improve the imaging sensitivity and reduce background signal of blood samples. Moreover, the antiepithelial-cell-adhesion-molecule (EpCAM) antibody-labeled magnetic nanospheres have been used for highly capture rare tumor cells in whole blood. The near-infrared nanoprobe with signal amplification and IMNs platform exhibits excellent performance in efficient capture and detection of CTCs, which shows great potential in cancer diagnostics and therapeutics.

Bioaccessibility of As and Pb in orchard and urban soils amended with phosphate, Fe oxide and organic matter.

Soils historically contaminated in urban and orchard environments by Pb and As were amended separately with organic matter, soluble Ca phosphate, and Fe oxide to determine whether these materials could lower Pb or As bioaccessibility. After 5 years of equilibration in the laboratory, the amended soils and control were tested for bioaccessibility using the standard physiologically based extraction test (PBET). Bioaccessibilities of Pb and As were not substantially reduced relative to the unamended controls after the 5-year period by any of the soil amendments. Gastric bioaccessibility (GB) of Pb was in all cases much greater than gastrointestinal bioaccessibility (GIB) regardless of soil treatment, whereas GB and GIB of As were similar in magnitude for all soils. Both GB and GIB of Pb were greater in the orchard than the urban soil. Electron microprobe investigations identified discrete particulate forms of Pb in the soils by elemental mapping, and energy dispersive spectrometry (EDS) revealed a frequent spatial association of Pb-rich particles with phosphorus. It is suggested that Pb-rich particles in anthropogenically contaminated soils resist chemical transformation into less labile forms despite thermodynamic favorability because of their low surface area and low solubility. This kinetic effect could explain the observed ineffectiveness of amendments in reducing metal bioaccessibility.

Bioaccessibility of Ba, Cu, Pb, and Zn in urban garden and orchard soils.

Exposure of young children to toxic metals in urban environments is largely due to soil and dust ingestion. Soil particle size distribution and concentrations of toxic metals in different particle sizes are important risk factors in addition to bioaccessibility of these metals in the particles. Analysis of particle size distribution and metals concentrations for 13 soils, 12 sampled from urban gardens and 1 from orchard found that fine particles (<105 µm) comprised from 22 to 66% by weight of the tested soils, with Ba, Cu, Pb and Zn generally at higher concentrations in the finer particles. However, metal bioaccessibility was generally lower in finer particles, a trend most pronounced for Ba and Pb. Gastric was higher than gastrointestinal bioaccessibility for all metals except Cu. The lower bioaccessibility of Pb in urban garden soils compared to orchard soil is attributable to the higher organic matter content of the garden soils.

[Effect of covalent functionalization on DNA cleavage activity and ROS formation of single-walled carbon nanotubes].

Biotoxicity of selected covalent functionalizaed single-walled carbon nanotubes (SWNTs) was investigated by plasmid DNA cleavage assay. The effect of covalent functionalization on the physical structural and chemical activity of SWNTs was also studied by Vis-NIR spectroscopy, Raman spectroscopy and ROS assay. The DNA cleavage intensity was promoted by covalent functionalization, and the intensity of DNA damage caused by the three types of carbon nanotubes in a descending order was found to be carboxylated SWNTs (SWNT-COOH) > polyethyleneglycol-lated SWNTs (SWNT-PEG) > uncovalent functionalized SWNTs (uSWNTs). SWNTs were efficient intermediate to trigger electron transfer from electron donor to O2 in dark condition which lead to superoxide radical (O2*-) formation. The electron transfer intensity caused by selected SWNTs was also SWNT-COOH > SWNT-PEG > SWNTs. The reason is that defect sites which changed from sp2-hybridized carbon atom on the side wall of carbon nanotubes were more reactive than the perfect ones.

The involvement of spinal bovine adrenal medulla 22-like peptide, the proenkephalin derivative, in modulation of nociceptive processing.

Bovine adrenal medulla 22 (BAM22), one of the cleavage products of proenkephalin A, possesses high affinity for opioid receptors and sensory neuron-specific receptor (SNSR). The present study was designed to examine the expression of BAM22 in the spinal cord and dorsal root ganglion (DRG) of naive rats as well as in a model of inflammation. BAM22-like immunoreactivity (BAM22-IR) was expressed in fibers in the spinal cord, with high density seen in lamina I in naïve rats. The expression of BAM22-IR in the superficial laminae was greatly reduced following dorsal rhizotomy. BAM22-IR was also located in 19% of DRG cells, mainly in the small- and medium-sized subpopulations. Following injection of complete Freund's adjuvant (CFA) in the hindpaw, the expression of BAM22-IR in the superficial laminae of the spinal cord and small-sized DRG neurons on the ipsilateral side was markedly increased. Double labeling showed that the Fos-positive nucleus was surrounded by BAM22-IR cytoplasm in the spinal dorsal horn neurons or closely associated with BAM22-IR fibers in the superficial laminae. Furthermore, CFA-induced mechanical allodynia in the inflamed paw was potentiated by intrathecal administration of anti-BAM22 antibody. Together, these results demonstrate for the first time that BAM22-like peptide is mainly located in the superficial laminae of the spinal cord and mostly originates from nociceptive DRG neurons. BAM22 could thus act as a ligand for presynaptic opioid receptors and SNSR. Our study also provides evidence suggesting that BAM22 plays a role in the modulation of nociceptive processing at the spinal level under normal and inflammatory conditions.

The passivation of pyrrhotite by surface coating.

The potential of triethylenetetramine (TETA) to inhibit the oxidation of three pyrrhotites, Garson, McCreedy and Po-97 has been studied systematically and confirmed by comparing the release of Fe and SO4(2-) from samples with and without coating treatment. Each sample, original or coated by TETA, was exposed to oxygen, 1 x 10(-3) M FeCl3, and Acidithiobacillus ferrooxidans, respectively, for specific oxidation periods. Both abiotic and biotic oxidation of samples treated by this passivating agent has been reduced significantly in this study. Under the aerobic condition, lower concentrations of ferric, total Fe or SO4(2-) were obtained from the coated samples than those from the uncoated samples. In the presence of 1 x 10(-3) M FeCl3 at 30 degrees C, TETA was able to reduce oxidation rates of Garson, McCreedy and Po-97 by 83%, 79%, and 81% (based on Fe release), respectively. A higher pH, lower Eh, and lower concentrations of total Fe and SO4(2-) were also observed in the biotic oxidation of coated Garson by Acidithiobacillus ferrooxidans. The protection of pyrrhotite surface from oxidant attack by TETA barrier and the alkaline property of this coating agent can be used to interpret the inhibition of oxidation.