Double protein directed synthesis of chemically etched sulfur doped quantum dots for signal "on-off-on" sensing of glutathione mediated by copper ions.

In this study, a novel "on-off-on" fluorescent probe was suggested for sensitive and selective assay of glutathione (GSH). The as-fabricated nanoswitch employs a Cu2+-sulfur quantum dot system (SQ-dots/Cu2+). The surface reactivity and water solubility of SQ-dots were improved through capping with egg white and bovine serum albumin proteins. The surface functional groups on the surface of double protein-protected SQ-dots enhanced the interaction with Cu2+ ions, resulting in the aggregation induced quenching of SQ-dots. Addition of GSH, a strong Cu2+ ion chelator, disassembles the large aggregates into relatively smaller ones, restoring the fluorescence emission of SQ-dots. Under optimized conditions, the fluorescence intensity was increased by increasing GSH amounts within the range of 0.13-550 µM with a detection limit (S/N = 3) of 0.04 µM. The SQ-dots/Cu2+ system was successfully applied for the detection of GSH in different matrices such as dietary supplements, human serum, and vegetable extract samples. The as-fabricated probe holds great potential for the synthesis of other functionalized SQ-dots for (bio) sensing.

Ratiometric sensing interface for glutathione determination based on electro-polymerized copper-coordinated molecularly imprinted layer supported on silver/porous carbon hybrid.

A novel molecularly imprinted ratiometric-based sensor was designed for highly selective and ultrasensitive electrochemical detection of glutathione (GSH). The sensor consists of porous carbon co-doped with nitrogen and sulfur formed on the surface of graphite electrode (N, S@PC/GE). Silver nanoparticles (Ag) were grown on the surface of N, S@PC/GE to improve the conductivity/surface area of the sensor and represent an internal reference signal for ratiometric response. The monomer (pyrrole-4-carboxylic acid, Py-COOH) was electro-polymerized on the surface of Ag/N, S@PC/GE in the presence of Cu (II) to form Cu-MIP@Ag/N, S@PC/GE. Addition of GSH decreased the signal of Ag at 0.18 V (oxidation of Ag) due to coordination complexation, while the signal response at 0.83 V (oxidation of Ag-GSH complex) was increased. Under optimum conditions, the ratio response (IGSH/IAg) was increased with increasing the concentration of GSH in the range of 0.01-500 nM with a detection limit (S/N = 3) of 0.003 nM. The electrochemical sensor exhibits many advantages including low LOD, high selectivity, good reproducibility, and satisfactory stability. The sensor was successfully applied to determine GSH in dietary supplements and human serum samples with recoveries % ranged from 97.4 to 101.8% and relative standard deviation % (RSD %) did not exceed 3.8%. This research paper introduces new information for the construction of molecular imprinted ratiometric-based electrochemical sensors for highly selective and sensitive detection of (bio) molecules.

Effect of low fluence diode laser irradiation on the hydraulic conductivity of perfused trabecular meshwork endothelial cell monolayers.

OBJECTIVE: To determine the effect of low-fluence diode laser irradiation upon the fluid perfusion characteristics of cultured human trabecular meshwork cell monolayers when placed in a specially designed testing apparatus and subjected to fluid flow driven by a hydrostatic pressure gradient. METHODS: Two experimental series were conducted. In the first series, six low-fluence diode laser irradiation experiments were conducted using cultured human trabecular meshwork cell monolayers grown on filter supports. Upon reaching a steady state perfusion condition at approximately 5.0 mmHg, monolayers were irradiated at fluencies ranging from 0.2619 to 0.8571 J/cm2 using a diode laser (lambda=810 nm). Perfusion and data collection continued for 45 minutes post-irradiation, after which the monolayers were tested to determine post-experimental viability. Hydraulic conductivity values were analyzed for post-irradiation response in 2.5-minute intervals, grouped by viability. In the second series, a total of six irradiated experiments and six simultaneous nonirradiated control experiments were conducted. Fluence values of 0.3571 J/cm2 (n=3) and 0.4286 J/cm2 (n=3) were used. Hydraulic conductivity values were analyzed for post-irradiation response in 2.5-minute intervals, grouped by irradiated vs. nonirradiated control groups. RESULTS: In the first series, analysis showed that the viable monolayers exhibited a statistically significant increase in hydraulic conductivity (p<0.001) from 10 minutes post-irradiation onward. The non-viable monolayers exhibited a statistically significant decrease in hydraulic conductivity. In the second series, irradiated groups showed a significant difference (p<0.001) from nonirradiated controls from 10 minutes post-irradiation onward. CONCLUSION: Low-fluence diode laser irradiation increases hydraulic conductivity in viable perfused TM cell monolayers when compared to baseline values or simultaneous nonirradiated controls while decreasing hydraulic conductivity in nonviable monolayers.