Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary "Y" junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL(-1)) with a linear range of 6 orders of magnitude (from 10 fg mL(-1) to 50 ng mL(-1)). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe.
Diabetes is associated with an increased risk of cardio-vascular disease. A decrease in the number and functionality of endothelial progenitor cells (EPCs) leads to reduced endothelial repair and the development of cardiovascular disease. The aim of the present study was to explore the effect and underlying mechanisms of nuclear factor erythroid 2-related factor 2 (Nrf2) on EPC dysfunction caused by diabetic mellitus. The biological functions of EPCs in streptozotocin-induced diabetic mice were evaluated, including migration, proliferation, angiogenesis and the secretion of vascular endothelial growth factor (VEGF), stromal-derived growth factor (SDF) and nitric oxide (NO). Oxidative stress levels in diabetic EPCs were also assessed by detecting intracellular reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA). EPC senescence was evaluated by measuring p16 and b-gal expression and observing the senescence-associated secretory phenotype. In addition, the function of EPCs and level of oxidative stress were assessed following Nrf2 silencing or activation. Nrf2 silencing resulted in a decrease of EPC biological functions, accelerated cell senescence and increased oxidative stress, as indicated by ROS and MDA upregulation accompanied with decreased SOD activity. Furthermore, Nrf2 silencing inhibited migration, proliferation and secretion in EPCs, while it increased oxidative stress and cell senescence. Nrf2 activation protected diabetic EPCs against the effects of oxidative stress and cell senescence, ameliorating the biological dysfunction of EPCs derived from mice with diabetes. In conclusion, Nrf2 overexpression protected against oxidative stress-induced functional damage in EPCs derived from diabetic mice by regulating cell senescence.
For the first time, hemin/G-quadruplex was employed to simultaneously serve as NADH oxidase and an HRP-mimicking DNAzyme for constructing a simple and sensitive pseudobienzyme-amplifying electrochemical aptasensor for thrombin detection.
A novel functionalized material is synthesized using surface-decorated fullerene (C60) to encapsulate hollow and porous palladium nanocages (PdNCs), and is applied to fabricate an electrochemiluminescence (ECL) immunosensor for the detection of Streptococcus suis Serotype 2 (SS2). PdNCs with hollow interiors and porous walls are prepared using a galvanic replacement reaction between silver nanocubes and metal precursor salts. Then, C60 reacts with L-cysteine (L-Cys) to form L-Cys functionalized C60 (C60-L-Cys), which has a better biocompatibility, conductivity, and hydrophilicity compared to C60 and possesses abundant -SH groups on the surface. Because of the special interaction between -SH and PdNCs, the obtained C60-L-Cys is adsorbed around the PdNCs to form an interesting structure with multiple spheres encapsulating the cage. The resultant functionalized material (C60 -L-Cys-PdNCs) has a high specific surface area, good electrocatalytic ability, and efficient photocatalytic activity, and is used to construct an ECL immunosensor for the detection of SS2. The ECL signal amplified strategy is performed by using the novel coreactant (C60-L-Cys) and in situ generation of O2 thus creating the S2O8(2-)-O2 ECL system. As a result, a wide linear detection range of 0.1 pg mL(-1) to 100 ng mL(-1) is acquired with a relatively low detection limit of 33.3 fg mL(-1).
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.