An optical DNA logic gate based on strand displacement and magnetic separation,with response to multiple microRNAs in cancer cell lysates

A battery of logic gates, “YES”, “AND” and “OR”, are constructed using magnetic beads (MBs) modified by DNA which consists of a substrate strand (S) and a signal strand on which the logic operates. Inputs stemming from micro-RNA (which represent three cancer biomarkers) take the place of signal DNA. The released signal strand self-assembles into the hemin-G-quadruplex complex (DNAzyme) that catalyzes a blue-green dye (ABTS⁺) from the precursor ABTS. This dye (quantified at a wavelength of 414 nm) represents the output signal for the various logic gates. The method allows quantitative detection of microRNA of three kinds of logic gates in the range of 5 nM–500 nM with detection limits of 3.8 nM, 4.9 nM, 5.4 nM. Boolean logic circuitry is also achieved following the principles of multilevel strand displacement. Based on strand displacement and magnetic separation, this work demonstrates the possibility of designing a logic system using micro-RNA in live cell lysate as inputs, and its potential application in DNA computation and cancer diagnosis.

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Graphical abstract Schematic representation of a battery of logic gates and the Boolean logic circuitry based on strand displacement and magnetic separation responding to multiple microRNA in cancer cell lysate.

     

Amperometric nitrite sensor based on a glassy carbon electrode modified with electrodeposited poly(3,4-ethylenedioxythiophene) doped with a polyacenic semiconductor

Microchimica Acta - The paper describes an electrochemical nitrite sensor based on the use of a nanocomposite consisting of poly(3,4-ethylenedioxythiophene) (PEDOT) doped with polyacenic...     

Voltammetric immunoassay for the carcinoembryonic antigen by using a glassy carbon electrode modified with poly(3,4-ethylenedioxythiophene) doped with tannic acid and grafted with poly(ethylene glycol)

The authors describe a voltammetric immunoassay for the carcinoembryonic antigen (CEA). A GCE was modified by electrodeposition of poly(3,4-ethylenedioxythiophene) (PEDOT) doped with tannic acid (TA). Subsequently, four-armed poly(ethylene glycol) (PEG) was assembled onto the modified surface through hydrogen bonding. The fabrication steps were characterized by scanning electron microscopy, energy dispersive spectroscopy, fourier transform infrared spectroscopy, contact angle measurements, electrochemical impedance spectroscopy and differential pulse voltammetry. The PEG/TA-PEDOT surface is shown be super-hydrophilic and to possess anti-fouling capability. Antibody against CEA was then covalently immobilized on the electrode. By using hexacyanoferrate as an electrochemical probe and at a working potential of 0.18 V vs SCE, the amperometric response is linear in the 10 ag·mL^?1 to 1.0 ng·mL^?1 CEA concentration range, and the detection limit is as low as 4.8 ag·mL^?1 (at an S/N ratio of 3). The assay was applied to the quantification of CEA in 1:10 diluted human serum samples. Recoveries ranged from 103.7 to 108.7%, and relative standard deviations from 2.9 to 4.8%.

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Graphical abstract Schematic of an electrochemical immunosensor for the carcinoembryonic antigen (CEA). It is based on the use of tannic acid (TA) and poly(ethylene glycol) (PEG), both deposited on a glassy carbon electrode (GCE), and using hexacyanoferrate as the electrochemical probe. The sensor has a wide linear range and a 4.8 ag·mL^?1 detection limit.

     

One-pot synthesis of tricyclo-1,4-benzoxazines via visible-light photoredox catalysis in continuous flow

A facile one-pot synthesis of tricyclo-1,4-benzoxazines has been developed via metal-free intramolecular cyclization of indole derivates. These reactions were efficiently achieved at ambient temperature by using visible-light photoredox catalysis in continuous flow. This directed intramolecular cyclization could be easily handled and scaled up in an open flask, enabling construction of a focused compound library for further pharmacological evaluation.     

Efficient P,O chelate palladium(II)/AgNO3 cocatalyzed homocoupling of aromatic terminal alkynes in aqueous media under ambient atmosphere

A new and efficient protocol for the P, O chelate Pd(II)/AgNO₃ cocatalyzed oxidative homocoupling reaction of aromatic terminal alkynes in the synthesis of symmetrical 1,4-disubstituted-1,3-diynes was described in aqueous media under ambient atmosphere. The results showed that both NEt₃ and THF/H₂O (in 4:1 proportion) played crucial roles in the reaction. In contrast, this protocol employs a low palladium(II) complex loading and AgNO₃ as cocatalyst to obtain the homocoupled products in moderate to good yields.     

Pulse broadening of the femtosecond pulses in a Gaussian beam passing an angular disperser

A general analytical formula has been found to describe the evolution of the pulse width of the femtosecond pulses in a Gaussian beam after passing an angular disperser without the assumption of well collimation. This formula is experimentally verified by measuring the pulse width with an autocorrelator based on two-photon absorption. It is found that the effect of the spectral lateral walk-off and group delay dispersion on the pulse-width evolution, and its dependence on the distance traveled, are substantially different when the beam has not been well collimated than from when it has been collimated. These differences result from the decaying nature of the angular dispersion of the Gaussian beam sent across a distance.     

Inverse temperature dependence of strain hardening in ultrahigh molecular weight polyethylene: role of lamellar coupling and entanglement density

Irradiation of ultrahigh molecular weight polyethylene (UHMWPE) with a dose of 150 kGy by an electron beam can effectively increase the entanglement density in the amorphous phase and has little influence on the properties of the crystalline phase, which provides examples to comparatively investigate the role of lamellar coupling and entanglement density in determining the strain-hardening effect in semicrystalline polymers. The strain-hardening modulus, deduced from the Haward plots of true stress-strain curves, is inversely temperature-dependent and has a sharp transition around 65 degrees C that corresponds to the mechanical alphaI-process of the crystalline phase for both nonirradiated and irradiated samples, irrespective of the entanglement density in the amorphous phase. Lamellar coupling takes more effect in determining the strain-hardening behavior before the mechanical alphaI-process is activated. With further increasing temperature, lamellar coupling becomes weaker and the role of the entangled amorphous phase is gradually presented. However, the same temperature dependence of the strain-hardening modulus in both nonirradiated and irradiated samples indicates that the strain-hardening behavior in semicrystalline polymer is mostly determined by lamellar coupling rather than by entanglement density.     

Synthesis, antiviral and antifungal bioactivity of 2-cyano-acrylate derivatives containing phosphonyl moieties

Alkyl 2-cyano-3-methylthio-3-phosphonylacrylates were synthesized by the reaction of alkyl 2-cyano-3,3-dimethylthioacrylates with dialkyl phosphites. The structures of the new compounds were characterized by elemental analyses, IR, 1H-, 13C- and 31P-NMR spectral data. These compounds were tested in vitro against pathogenic fungi, namely, Fusarium graminearum, Cytospora mandshurica and Fusarium oxysporum. Amongst all compounds, 2d and 2t were found to be effective against the tested fungi at 50 microg/mL. A half-leaf method was used to determine the in vivo protective, inactivation and curative efficacies of the title products against tobacco mosaic virus (TMV). Title compounds 2a and 2b were found to possess good in vivo curative, protection and inactivation effects against TMV with inhibitory rates at 500 mg/L of 60.0, 89.4 and 56.5 and 64.2, 84.2 and 61.2 %, respectively. To the best of our knowledge, this is the first report on the antiviral and antifungal activity of alkyl 2-cyano-3-methylthio-3-phosphonylacrylates.     

Quantitative determination of soybean meal content in compound feeds: comparison of near-infrared spectroscopy and real-time PCR

Standard methods for determining the raw material content of compound feed are little exploited, except for the identification of meat and bone meal in feeds. In this work, near-infrared (NIR) spectroscopy and real-time polymerase chain reaction (PCR) were applied in order to establish new and fast methods for quantification of soybean meal content in compound feeds. The best prediction quality was achieved by using a model based on NIR spectroscopy (R ² = 0.9857, standard error of cross-validation 1.1065). Furthermore, a sensitive qualitative detection method by using the real-time PCR was developed (R ² = 0.976, slope −3.7599). Finally, the differences between the real-time PCR result and the NIR spectroscopy result for a given sample were also treated, and we found that the NIR spectroscopy method provided quite accurate results which approach closely those of the real-time PCR method. Hui Li and Xiaowen Lv contributed equally to this work.     

At-line coupling of UPLC to chip-electrospray-FTICR-MS

Since highly sensitive on-line coupling of UPLC with FTICR-MS is technically infeasible due to their different scan rates, at-line coupling of these techniques was developed for rapid analysis. To enable cutting of one peak of the chromatogram into one fraction, several conditions and relationships were investigated, e.g. the optimum volume of the inserted delay loop, the relationship between retention time, loop outlet drop speed, individual drop volume versus mobile phase composition under constant speed, and linear solvent strength gradient elution modes. Good and reproducible results were achieved applying UPLC as an efficient separation and fast fractionation tool before the FTICR-MS measurements. A chip-based nanoelectrospray ionization system was employed which was perfectly suited to handling the small-volume fractions and was thus chosen for the at-line coupling. The method was initially applied to spiked extracts of cell-free bacterial culture supernatants in which bacterial signalling compounds, namely N-acyl homoserine lactones (AHL), were detected. Good reproducibility and high recovery was observed. Afterwards, a culture supernatant of Erwinia sp. JX3.2, a putative AHL producer, was investigated and N-hexanoyl-homoserine lactone was determined as a possible signalling molecule. More reliable assignments were achieved by use of at-line coupling of UPLC and FTICR-MS compared with off-line measurements. Xiaojing Li and Agnes Fekete have contributed equally to this publication.