Carbon dot enhanced peroxidase-like activity of platinum nanozymes.

As one of the most intriguing nanozymes, the platinum (Pt) nanozyme has attracted tremendous research interest due to its various catalytic activities but its application is still limited by its poor colloidal stability and low affinity to substrates. Here, we design a highly stable Pt@carbon dot (Pt@CD) hybrid nanozyme with enhanced peroxidase (POD)-like activity (specific activity of 1877 U mg-1). The Pt@CDs catalyze the decomposition of hydrogen peroxide (H2O2) to produce singlet oxygen and hydroxyl radicals and exhibit high affinity to H2O2 and high specificity to 3,3',5,5'-tetramethyl-benzidine. We reveal that both the hydroxyl and carbonyl groups of CDs could coordinate with Pt2+ and then regulate the charge state of the Pt nanozyme, facilitating the formation of Pt@CDs and improving the POD-like activity of Pt@CDs. Colorimetric detection assays based on Pt@CDs for H2O2, dopamine, and glucose with a satisfactory detection performance are achieved. Moreover, the Pt@CDs show a H2O2-involving antibacterial effect by destroying the cell membrane. Our findings provide new opportunities for designing hybrid nanozymes with desirable stability and catalytic performance by using CDs as nucleating templates and stabilizers.

Nucleic Acid Hybridization Enhanced Luminescence for Rapid and Sensitive RNA and DNA Based Diagnostics.

Long-lived emissive nucleic acid probes are widely used in biochemical analysis due to their programmable structures, high signal-to-background ratio, and high sensitivity. Homogeneous detection based on long-lived emissive nucleic acid probes is often achieved through Förster resonance energy transfer (FRET), which suffers from the limitation of a narrow effective distance range. Herein, a new strategy of accessing nucleic acid hybridization-responsive luminescent probes is presented. The photoluminescence (PL) of a Lumi4-Tb complex internally modified with DNA is switched on by nucleic acid hybridization, after which the PL is increased up to 20 times. PL lifetime analysis revealed a possible mechanism of luminescence enhancement. Due to the flexibility of single-stranded nucleic acid chains, the bases and phosphate groups can coordinate with the Tb(III), which reduces the stability of the Tb complex and results in weak PL. After hybridization, the rigid double helix structure suppresses the coordination between Tb(III) and the bases or phosphate groups, causing luminescence enhancement. As the DNA sequence can be freely designed, an array of probes for different DNA or RNA targets can be created with the same Tb complex. Moreover, the novel probe design can afford pM detection limits of DNA or RNA without any nucleic acid amplification and exhibits great potential for nucleic acid detection in clinical diagnosis.

Quantitatively Switchable pH-Sensitive Photoluminescence of Carbon Nanodots.

pH sensing plays a key role in the life sciences as well as the environmental, industrial, and agricultural fields. Carbon nanodots (C-dots) with small size, low toxicity, and excellent stability hold great potential in pH sensing as nanoprobes due to their intrinsic pH-sensitive photoluminescence (PL). Nonetheless, the undesirable sensitivity and response range of C-dot PL toward pH cannot meet the requirements of practical applications, and the unclear pH-sensitive PL mechanism makes it difficult to control their pH sensitivity. Herein, the quantitative correlation of pH-sensitive PL with specific surface structures of C-dots is uncovered for the first time, to our best knowledge. The association of carboxylate and H+ increases the ratio of nonradiation to radiation decay of C-dots through excited-state proton transfer, resulting in the decrease of PL intensity. Meanwhile, the dissociation of α-H in ß-dicarbonyl forming enolate increases the extent of delocalization of the C-dots conjugated system, which induces the PL broadening to the red region and a decreasing intensity. Based on the understanding of the pH-sensitive PL mechanism, the pH-sensitive PL of C-dots can be switched by quantitative modulation of carboxyl and ß-dicarbonyl groups to achieve a desirable pH response range with high sensitivity. This work contributes to a better understanding of the pH-sensitive PL of C-dots and therefore presents an effective strategy for controllably tuning their pH sensitivity, facilitating the rational design of C-dot-based pH sensors.

Novel liquid fermentation medium of Cordyceps militaris and optimization of hydrothermal reflux extraction of cordycepin.

In this study, we developed a novel liquid fermentation medium of Cordyceps militaris using pupa powder and wheat bran as nitrogen resources instead of the traditionally used peptone. This process not only reduced the cost by approximately 50%, but increased production by over 30%. Then, we explored a method to extract and purify cordycepin by combining hydrothermal reflux extraction with macroporous resin adsorption, which is inexpensive and suitable for the industrial production. The optimum conditions for hydrothermal reflux were extracting three times at 95 °C with 1:10 sample-to-water ratio, and the cordycepin purity with macroporous resin HPD-100 reached 95.23%.[Formula: see text].

Molecular Tweezers-like Calix[4]arene Based Alkaline Earth Metal Cation (Ca2+, Sr2+, and Ba2+) Chemosensor and Its Imaging in Living Cells and Zebrafish.

Although alkaline earth metal cations play an important role in our daily life, little attention has been paid to the field of fast quantitative analysis of their content due to a lack of satisfactory precision and a fast and convenient means of detection. In this study, we have designed a set of molecular tweezers based on the calix[4]arene chemosensor L, which was found to exhibit high selectivity and sensitivity toward Ca2+, Sr2+, and Ba2+ (by UV-vis and fluorescence methods) with low detection limits of the order of 10-7 to 10-8 M and high association constants (of the order of 106). More significantly, sensor L not only can recognize Ca2+, Sr2+, and Ba2+ but also can further discriminate between these three cations via the differing red shifts in their UV-vis spectra (560 nm for L·Ca2+, 570 nm for L·Sr2+, and 580 nm for L·Ba2+ complex) which is attributed to their different atomic radii. A rare synergistic effect for the recognition mechanism has been demonstrated by 1H NMR spectroscopic titration. Sensor L constructed a high shielding field by the cooperation of Tris with alkaline earth metal ion after complex. Additionally, the presence of acetoxymethyl group in sensor L results in enhancement of cell permeability, and as a consequence, sensor L exhibited excellent sensing and imaging (in vivo) in living cells and in zebrafish.

An efficient ICT-based ratio/colorimetric tripodal azobenzene probe for the recognition/discrimination of F-, AcO- and H2PO4- anions.

The tripodal probe L was readily prepared via introducing rhodamine and azobenzene groups in a two-step procedure. Studies of the recognition properties indicated that probe L exhibited high sensitivity and selectivity towards F-, AcO- and H2PO4- through a ratiometric colorimetric response with low detection limits of the order of 10-7 M. The complexation behaviour was fully investigated by spectral titration, 1H NMR spectroscopic titration and mass spectrometry. Probe L not only recognizes F-, AcO- and H2PO4-, but can also distinguish between these three anions via the different ratiometric behaviour in their UV-vis spectra (387/505 nm for L-H2PO4-, 387/530 nm for L-AcO- and 387/575 nm for L-F- complex) or via different colour changes (light coral for L-H2PO4-, light pink for L-AcO- and violet for the L-F- complex). Additionally, given the presence of NH and OH groups in probe L, which can be protonated and deprotonated, probe L further exhibited an excellent pH response over a wide pH range (pH 3 to pH 12).

Recognition of silver cations by multifarene[2,2] chemosensors with unexpected fluorescence response.

Fluorescent chemosensors based on a new macrocyclic compound, multifarene[2,2], with modification by triazole-linked pyrene or anthracene were synthesized. These macrocyclic sensors exhibited high affinity and selectivity toward Ag+ over other metal ions, with ratiometric or enhanced response of their fluorescence emissions depending upon the substituent species for coordination to Ag+, and an unexpected response to a concentration threshold of the metal cations was discovered. The experimental evidences of fluorescence spectra, 1H NMR titration, IR spectra, and high-resolution mass spectra suggested the coordination behaviors of the sensors with Ag+, that is, the 1:1 complexes were formed with moderate association constants of about 105 L·mol-1, and the sulfur atoms on macrocyclic ligand should affinite to the metal cations. Energy-minimized structures and frontier orbitals were estimated by quantum chemical calculations with a view to rationalizing the fluorescence response of the multifarene[2,2] sensors upon binding to Ag+.

Properties comparison between free and immobilized wheat esterase using glass fiber film.

In this paper, the catalytic performance of non-purified esterase from wheat bran immobilized on glass fibre membrane carrier is established, the immobilization conditions observed were enzyme 1 mL, phosphate buffer 3 mL (pH 7.0), immobilization time 1 h, immobilization temperature 29 °C. After carrier functionalization some characteristics of immobilized enzyme were studied, the results showed that immobilized enzyme presenting improved characteristic than that of free enzyme. The optimum pH for free and immobilized enzymes were found to be 8 and 7, respectively. As for optimum temperature for free and immobilized enzymes were observed to be 30 °C and 40 °C, respectively. When the enzyme was immobilized on glass fibre membranes, its Km increased about 7 times. In addition, storage and thermal stability of the free wheat esterase were increased by as a result of membrane immobilization, after 12 days of storage, the immobilized enzyme still retained about 91.10% of its original activity at 4 °C, indicating a great potential in industrial application.

Chemical composition and antimicrobial activities of volatile oil extracted from Chrysanthemum morifolium Ramat.

Volatile oil in Chrysanthemum morifolium Ramat (C. morifolium) was extracted by the method of water vapor distillation and its chemical components was identified by gas-chromatography coupled with mass spectrometry (GC-MS). The volatile oil are evaluated for antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella enteritids, Pseudomonas aeruginosa and Bacillus subtilis. Effects of surfactant, temperature, pH and ultraviolet light on antibacterial activity stability of volatile oil were analyzed too. Total 56 compounds were identified in C. morifolium volatile oil. The main constituents in C. morifolium volatile oil were monoterpenes and sesquiterpenes compounds, including hydrocarbons, esters, aldehydes, ketones, phenols and organic acids. α-curcumene was the most abundant volatile component (12.55%). The volatile oil showed promising antibacterial activity against 5 selected strains. The inhibitory effect on P. aeruginosa exhibited maximum inhibition zone diameter 20.43 mm, and E. coli showed 12.29 mm. The volatile oil treated with surfactant Tween 20 showed the strongest antibacterial activity, followed by Tween 80 and the SDS lowest, which showed the lowest. pH also had different effect on antibacterial activity stability of the C. morifolium volatile oil. No significant difference effect on antibacterial activity stability of volatile oil was observed with temperature and UV treatment.

[4-amino-1,8-naphthalimide on the Sensitive effect of arsenic trioxide in hepatocellular carcinoma cells].

OBJECTIVE: To study the effect and mechanism of 4-amino-1, 8-naphthalimide (4-AN) on the sensitive effect of arsenic trioxide (ATO) in hepatocellular carcinoma cells. METHODS: Hepatocellular carcinoma HepG2 cells were divided into two groups according to whether they were treated with 4-AN or not. Cell viability was evaluated by MTT assay, population doubling experiment and colony formation assay; genic mechanism was explored by 8-OH-dG assay, single cell gel electrophoresis (comet assay) and microriucleus test. RESULTS: At 2-10 micromol/L concentration of ATO, the cell viability and colony formation efficiency of the combinatio group (4-AN+ATO) were significantly lower than that of the ATO group (P<0.05); moreover, the tail-length (L-Tail) and olive tail moment (OTM) in comet assay were notablely higher than that of the ATO group (P<0.05). At 2-20 micromol/L concentration of ATO, the population doubling time and 8-OH-dG in combination group were significantly higher than that of ATO group (P<0.05). Results from DNA damage repair assay showed that the efficiency of DNA damage repair in combination group was remarkably lower than that of ATO group (P<0.05). At 5-20 micromol/L concentration of ATO, the frequency of micronucleated cells in combination group was significantly higher than that of ATO group (P<0.05). CONCLUSION: 4-AN can significantly increase the sensitivity of ATO in treatment with hepatocellular carcinoma cells and prevent DNA damage repair may be a primary mechanism for this effect.

An electrochemical and surface plasmon resonance study of adsorption actions of DNA by Escherichia coli.

In this work, we proposed an immobilization scheme targeting Escherichia coli bacterial cells onto gold surface utilizing polypeptide RGD's binding specificity to gold particles. Adsorption kinetics of extracellular Herring Sperm DNA on E. coli was then studied using electrochemical methods along with surface plasmon resonance spectroscopy through this immobilization scheme. The adsorption equilibrium constants of DNA adsorbing to E. coli from electrochemical method and surface plasmon resonance spectroscopy, were (5.596±0.462)×105 L mol(-1) and (1.24±0.361)×105 L mol(-1), accordingly. Importantly, this is the first study that used an electrochemical method to express the adsorptive action of DNA by E. coli.

Evaluation of nonspecific interactions between quantum dots and proteins.

A method based on the AFM and colloidal probe techniques was proposed to directly measure nonspecific interactions between QDs and different proteins with respective sizes and isoelectric points. Results indicated that van der Waals forces were the leading force, while electrostatic interactions also played an important role in nonspecific interactions.

Nanomechanical analysis of yeast cells in CdSe quantum dot biosynthesis.

QD biosynthesis affects the mechanical strength of yeast cells. The intracellular synthesis of CdSe QD in yeast cells incubated with Na2 SeO3 and subsequently with CdCl2 increases the glucan content of their cell walls, resulting in their enhanced mechanical strength.

Graphene oxide based fluorescent aptasensor for adenosine deaminase detection using adenosine as the substrate.

We present a novel fluorescent aptasensor for simple and accurate detection of adenosine deaminase (ADA) activity and inhibition on the basis of graphene oxide (GO) using adenosine (AD) as the substrate. This aptasensor consists of a dye-labeled single-stranded AD specific aptamer, GO and AD. The fluorescence intensity of the dye-labeled AD specific aptamer is quenched very efficiently by GO as a result of strong π-π stacking interaction and excellent electronic transference of GO. In the presence of AD, the fluorescence of the GO-based probe is recovered since the competitive binding of AD and GO with the dye-labeled aptamer prevents the adsorption of dye-labeled aptamer on GO. When ADA was introduced to this GO-based probe solution, the fluorescence of the probe was quenched owing to ADA can convert AD into inosine which has no affinity to the dye-labeled aptamer, thus allowing quantitative investigation of ADA activity. The as-proposed sensor is highly selective and sensitive for the assay of ADA activity with a detection limit of 0.0129U/mL in clean buffer, which is more than one order of magnitude lower than the previous reports. Meanwhile, a good linear relationship with the correlation coefficient of R=0.9922 was obtained by testing 5% human serum containing a series of concentrations of ADA. Additionally, the inhibition effect of erythro-9-(2-hydroxy-3-nonyl) adenine on ADA activity was investigated in this design. The GO-based fluorescence aptasensor not only provides a simple, cost-effective and sensitive platform for the detection of ADA and its inhibitor but also shows great potential in the diagnosis of ADA-relevant diseases and drug development.

One-to-one quantum dot-labeled single long DNA probes.

Quantum dots (QDs) have been received most attention due to their unique properties. Constructing QDs conjugated with certain number of biomolecules is considered as one of the most important research goals in nanobiotechnology. In this study, we report polymerase chain reaction (PCR) amplification of primer oligonucleotides bound to QDs, termed as QD-based PCR. Characterization of QD-based PCR products by gel electrophoresis and atomic force microscopy showed that QD-labeled long DNA strands were synthesized and only a single long DNA strand was conjugated with a QD. The QD-based PCR products still kept fluorescence properties. Moreover, the one-to-one QD-labeled long DNA conjugates as probes could detect a single-copy gene on maize chromosomes by fluorescence in situ hybridization. Labeling a single QD to a single long DNA will make detection of small single-copy DNA fragments, quantitative detection and single molecule imaging come true by nanotechnology, and it will promote medical diagnosis and basic biological research as well as nano-material fabrication.

[Effects of pol beta on biological characteristics and DNA damage in mouse embryonic fibroblast].

OBJECTIVE: To explore the effect of DNA polymerase beta (pol beta) expression level on biological characteristics of mouse embryonic fibroblast (MEF) and the cellular response to DNA damage induced by potassium dichromate. METHODS: pol beta wild-type cells (pol beta +/+), pol beta null cells (pol beta -/-) and pol beta overexpressed cells (pol beta oe) were applied as a model system. The growth curve of cells was plotted by MTT assay; the doubling time of cells was detected by double time experiment; the spontaneous mutation frequency was determined by HGPRT gene mutation method and single cell gel electrophoresis assay (SCGE) was employed to observe the DNA damage either happened spontaneously or induced by potassium dichromate. RESULTS: Growth characteristic and doubling time of the three kinds of cells were similar and no obvious differences were found on spontaneous DNA damage and mutations frequency among them (P > 0.05). Potassium dichromate increased comet rate and tail length in the three kinds of cells in a concentration dependent way. DNA damage of pol beta -/- cells at the same dosage were more serious than the other cells both in comet rate and tail length (P < 0.05). pol beta oe cells demonstrated more resistant to DNA damage obviously than the others. CONCLUSION: The expression level of pol beta has no significant effect on the biological characteristic and spontaneous mutation frequency of MEF. pol beta knock out cells is more sensitive to DNA damage induced by potassium dichromate, whereas, pol beta over expression can help cells response to DNA damage and protect cells from death in a certain degree.

[Effect of metabolism activation on the oxidative cell damage induced by cigarette smoking].

OBJECTIVE: To explore the effect of metabolic activation on the oxidative cell damage induced by mainstream smoke (MS). METHODS: A549 cells were treated with different concentrations of MS for 2 hours in the presence and absence of S9 mix. ROS levels were determined. DNA and chromosome damages were detected. The activities of SOD and GSH-Px were measured. RESULTS: With different concentrations of MS treatment, the contents of ROS in the cells with S9 mix were significantly higher than those without S9 mix (P<0.05). More serious chromosome and DNA damages in the cells with S9 mix were observed than those without S9 mix. The micronucleus rate, comet rate, L Tail and OTM in the cells with S9 mix were significantly higher than those without S9 mix (P<0.05). With increased dose of MS, the activities of SOD and GSH-Px decreased in both cells with and without S9 mix. But the tendency of decrease in the cells with S9 mix was more obvious than those without S9 mix (P<0.05). CONCLUSION: MS induces oxidative damage in A549 cells and the metabolic activations could increase the oxidative stress and potentiate the cellular genotoxicity.

Surface manipulation for improving the sensitivity and selectivity of glassy carbon electrodes by electrochemical treatment.

The performance of electrochemical sensor is related to the surface structure of electrode. Electrochemical treatment was often used to improve the performance of glassy carbon electrodes (GCEs) for the simplicity of operation. The densely compact oxide film formed on the surface of the electro-oxidized glassy carbon electrodes (GCEs-ox) could be changed to be porous by following electro-reduction of the GCEs-ox obtaining the totally reduced GCEs-ox (GCEs-re). The GCEs-re with a porous film exhibited high sensitivity in detecting epinephrine (EP) accompanying with poor stability and incapability of anti-interference from negatively charged ascorbic acid (AA) because both of AA and EP could permeate the porous film. Compared to the GCEs-re, the GCEs-ox with a densely compact film displayed relatively lower sensitivity but higher stability. To combine the advantages of both porous and densely compact oxide films, a partially reduced GCE-ox (GCE-ox-re) with an oxide film porous outer and densely compact inner was designed by controlling the electro-reduction time and potential, which will provide a new strategy for improving the performance of GCEs. The GCEs-ox-re exhibited high sensitivity, excellent stability and high selectivity in the detection of EP. In the presence of 0.1 mM AA, the anodic peak current of EP was directly proportional to its concentration in the range of 8.0x10(-9)-2.0x10(-7) M and EP as low as 2.0x10(-9) M could be detected.

Tumor cell targeting using folate-conjugated fluorescent quantum dots and receptor-mediated endocytosis.

BACKGROUND: Luminescent nanobioprobes with cell-targeting specificity are likely to find important applications in bioanalysis, biomedicine, and clinical diagnosis. Quantum dots (QDs) are unique and promising materials for such a purpose because of their fluorescence and large surface area for attaching cell-targeting molecules. METHODS: We produced water-dispersible QDs by coating hydrophobic QDs with small amphiphilic polyethylene glycol (PEG) molecules via hydrophobic interactions. We covalently coupled folate (FA) onto the water-dispersible PEG-coated QDs (PEG-QDs) to produce FA-coupled PEG-QDs (FA-PEG-QDs). RESULTS: These FA-PEG-QD nanoparticles functioned as fluorescent nanobioprobes that specifically recognized folate receptors (FRs) overexpressed in human nasopharyngeal cells (KB cells) but not in an FR-deficient lung carcinoma cell line (A549 cells). Using confocal fluorescence microscopy, we demonstrated uptake of FA-PEG-QDs by KB cells but no uptake of folate-free PEG-QDs. The specificity of this receptor-mediated internalization was confirmed by comparing the uptake by KB vs A549 cells. CONCLUSIONS: Our results suggest that such cell-targeting fluorescent nanobioprobes are potentially very powerful tools for recognizing target cells and delivering and tracking drugs and other therapeutic materials.

[Oxidative damage of gasoline engine exhausts to rat lung tissues].

OBJECTIVE: To study the effects of extracts of condensate, particulates and semivolatile organic compounds from gasoline engine exhaust on DNA damage, 8-oxoguanine DNA glycosylase-1 (OGG1) expression, and changes of ultra-structures in lungs of rats. METHODS: Organic extracts of gasoline engine exhaust (GEE) was intratrachealy instilled into rat lungs at 0, 5.6, 16.7, and 50.0 L/kg body weight, respectively, once a week for a month. The single DNA strand break was measured by comet assay. The OGG1 was determined using immunohistochemistry method. The ultrastructure of lung cells was observed with electronic microscope. RESULTS: The rates of tailed cells detected by the comet assay increased significantly when the rats were exposed to 16.7 and 50.0 L/kg of GEE compared with those exposed to solvent only (P < 0.05). However, the tail length did not differ significantly between the groups. Similarly, exposure to 16.7 and 50.0 L/kg of GEE led to increased OGG1 significantly. Significant changes of mitochondria in type I and II alveolar cells as well as respiratory bronchiole epithelial cells were observed, which included decrease of numbers, pyknosis and swelling. CONCLUSION: Gasoline engine exhausts induce single DNA strand break, increase OGG1 expression, decrease numbers of mitochondria, and destroy ultrastructures of mitochondria in various lung cells of rats.