DNA Nanotweezers for Biosensing Applications: Recent Advances and Future Prospects.

DNA nanotweezers (DTs) are reversible DNA nanodevices that can optionally switch between opened and closed states. Due to their excellent flexibility and high programmability, they have been recognized as a promising platform for constructing a diversity of biosensors and logic gates, as well as a versatile tool for molecular biology studies. In this review, we provide an overview of biosensing applications using DTs. First, the design and working principle of DTs are introduced. Next, the signal producing principles of DTs are summarized. Furthermore, biosensing applications of DTs for varying targets and purposes, both in buffers and complex biological environments, are highlighted. Finally, we provide potential opportunities and challenges for the further development of DTs.

Binding induced isothermal amplification reaction to activate CRISPR/Cas12a for amplified electrochemiluminescence detection of rabies viral RNA via DNA nanotweezer structure switching.

Rabies is caused by the infection of Rabies virus, it leads to fatal encephalitis, developing a highly sensitive and specific detection method for Rabies virus remains a challenge. Herein, we report an electrochemiluminescence (ECL) biosensor for Rabies viral RNA based on dual-signal amplification and DNA nanotweezers (DTs). Dual-signal amplification process includes target binding induced isothermal amplification and CRISPR-based amplification. In the presence of target RNA, two assisted probes simultaneously hybridized with it to trigger isothermal amplification with the help of polymerase and nicking enzyme. This process generated a large amount of single-stranded DNA (ssDNA) as products. The products hybridized with CRISPR RNA to activate the trans-cleavage activity of Cas12a to indiscriminately cleave predesigned single-stranded trigger (ST) strands. After mixing the cleavage products with DTs and hemin molecules, DTs cannot be closed by cleaved ST strands to capture hemin to the electrode to quench the ECL signal. Therefore, the higher concentration of the target, the stronger intensity of the ECL signal. The detection limit is as low as 2.8 pM and the detection range is from 5 pM to 5 nM with excellent specificity and stability. The proposed method provides a promising strategy for Rabies detection, and can be easily adapted to other analytes via reasonable design as a valuable and versatile tool in bioanalysis.

[Production of antioxidative exopolysaccharides of Cordyceps militaris with Vernonia amygdalina leaves in substrate].

Cordyceps militaris exopolysaccharides (EPS) have many pharmacological activities such as boosting immunity and antifatigue. To obtain EPS efficiently, we added moderate Vernonia amygdalina leaf powder as inducer to the fermentation medium to promote the production of Cordyceps militaris EPS and studied the infrared absorption spectrum and antioxidant activities of the EPS after optimization. The optimum liquid fermentation conditions were as follows: addition of Vernonia amygdalina leaf powder of 8 g/L, fermentation duration of 9 d, initial pH of 6.5, inoculation quantity of 5.0 mL. Under such a condition, the yield of Cordyceps militaris EPS reached (5.24±0.28) mg/mL, increased by 205.20% compared to the control group without adding Vernonia amygdalina leaf powder. Results of infrared analysis and antioxidant activity showed that the Vernonia amygdalina leaves had little effect on the structure and activities of Cordyceps militaris EPS. The results of this research suggest that Vernonia amygdalina leaf can enhance the production of Cordyceps militaris EPS effectively, and provides a novel method for efficient production of EPS in Cordyceps militaris.

Characterization of the physicochemical properties and extraction optimization of natural melanin from Inonotus hispidus mushroom.

In this study, solid-state fermentation was used to produce Inonotus hispidus melanin (IH melanin). Its physicochemical properties were characterized, and ultrasonic-assisted extraction was used to optimize its extraction process. Characterization techniques like elemental analysis and so on were used to identify melanin. The solubility, stability, and antioxidant activity were also measured. Furthermore, the extraction process of IH melanin was optimized. The results showed that the pigment could be defined as dihydroxy phenylalanine (DOPA)-melanin, it displayed irregular spherical and ellipsoidal structures with average size of 89.33 nm. Melanin has a specific stability and shows antioxidant activity. The optimal extraction parameters of melanin were a NaOH concentration of 0.56 mol/L, solid-liquid ratio of 1:50, ultrasonic power of 300 W, extraction temperature of 70 °C, and ultrasonic time of 70 min. This study was the first time that IH melanin was obtained. The results may be applied to health food or food additives.

Characterization of natural melanin from Auricularia auricula and its hepatoprotective effect on acute alcohol liver injury in mice.

This study characterized the natural melanin from Auricularia auricula and investigated its hepatoprotective effect on mice with acute alcoholic liver injury. The characterization of the melanin was analyzed based on elemental analysis, gel permeation chromatography (GPC), UV-visible spectroscopy (UV-visible), infrared spectrum (IR) and nuclear magnetic resonance spectra (NMR). To determine the liver protective effect of Auricularia auricula melanin, mice were administered with the melanin once daily for 3 weeks before ethanol induced liver injury. Biochemical parameters of liver function, histopathological sections, mRNA and protein expression of antioxidant enzyme were compared between mice with or without the melanin administered. Results showed that A. auricula melanin was a eumelanin and the average molecular weight was 48.99 kDa. The melanin can protect the mice from ethanol-induced liver injury by extending the duration of the righting reflex, and shortening the duration of the recovery. The liver index, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), γ-glutamyl transpeptidase (γ-GT) and liver malondialdehyde (MDA) levels in mice treated with the melanin were significantly decreased. At the same time, the levels of liver alcohol dehydrogenase (ADH), and antioxidase such as catalase (CAT), and superoxide dismutase (SOD) were increased. Its protective effect may be related to the activation of nuclear factor E2-related factor 2 (Nrf2) and its downstream antioxidant enzymes such as glutamate cysteine ligase catalytic (GCLC), glutamate cysteine ligase modifier (GCLM), and NADP(H) quinine oxidoreductase 1 (NQO-1). These results suggested that A. auricula melanin may be an effective strategy to alleviate alcohol-induced liver damage.