Magnetic Solid-Phase Extraction of Fipronil from Aqueous Solution and Eggshells Using Amine-Coated Iron Oxide Nanoparticles.

Fipronil, as a broad use insecticide, is regulated under directives and standards in many countries since it is classed as Class II moderately hazardous pesticide and Group C possible human carcinogen. In this study, amine coated iron oxide (NH2 -Fe3 O4 ) is used as a new adsorbent with high adsorption capacity in the removal of fipronil from aqueous solution and eggshells.The adsorption performance was studied through batch adsorption under different concentrations of fipronil (10-200 µg/mL), different nanoparticles amount (0.1-1 mg) at different pH (4.5-6.0) and temperature (25-75 °C). Results showed that NH2 -Fe3 O4 nanoparticles (0.1 mg) have excellent adsorption efficiency (97.06 %) at 25 °C, and pH 5.5. It also showed higher adsorption capacity towards fipronil sulfide, fipronil sulfone and fipronil desulfinyl with removal efficiencies of 92.82 %, 86.35 % and 76.24 % from aqueous solution and 97.62 %, 76.97 % and 62.65 % from eggshells, respectively. Fipronil adsorption process on NH2 -Fe3 O4 nanoparticles showed best fitting to Langmuir adsorption isotherm indicating monolayer chemical adsorption by physicochemical interactions spontaneously on homogenous surfaces. Having higher adsorption capacity and reusability NH2 -Fe3 O4 nanoparticles were shown to be effective adsorbents for removal of fipronil from aqueous solution and eggshells.

Simultaneous Dual-Sensing Platform Based on Aptamer-Functionalized DNA Hydrogels for Visual and Fluorescence Detection of Chloramphenicol and Aflatoxin M1.

In this study, a chloramphenicol and aflatoxin M1 aptamer-functionalized DNA hydrogel was designed for the simultaneous detection of chloramphenicol and aflatoxin M1 for the first time. The acrydite-modified chloramphenicol aptamer sequence was used to synthesize the DNA hydrogel and for visual detection of chloramphenicol depending on the gel-to-sol transition of the target-responsive DNA hydrogel. The DNA hydrogel formulation was set as follows: 60% of each linear polyacrylamide-DNA conjugate and 40% of acrylamide and chloramphenicol aptamer/DNA strand-1 at a molar ratio of 1:1, and the lowest concentration of chloramphenicol leading to gel dissociation was 1.0 nM at 25 °C. Furthermore, the formalized aptamer-functionalized DNA hydrogel was used to detect aflatoxin M1 by measuring the recovery of the fluorescence signal that was quenched when the FAM-labeled aflatoxin M1 aptamer and BHQ1-labeled DNA strand-2 were hybridized to form a double-stranded DNA in the network of hydrogel. The detection platform was successfully applied to the detection of chloramphenicol and aflatoxin M1, both in aqueous solution and in milk. The aptamer-functionalized DNA hydrogel had detection (LOD) and quantification limits (LOQ) for aflatoxin M1 as 1.7 and 5.2 nM, respectively. Using two aptamer sequences with high affinity and specificity, the dual-sensing platform based on the DNA hydrogel achieved higher selectivity for chloramphenicol and aflatoxin M1, which demonstrated its potential as a reliable simultaneous detection platform against two different targets for monitoring food safety.

Adsorption behavior of carboxy- and amine-terminated magnetic beads for patulin: Batch experiments in aqueous solution and apple juice.

Patulin (PAT) is a highly water soluble, heat resistant and toxic fungal metabolite mostly contaminating apple juice. Due to its serious health effects, its removal from foodstuffs is required to ensure food safety. In this study, carboxy- and amine-terminated iron oxide spheres (Fe3O4-COOH and Fe3O4-NH2) were investigated for their adsorbing capacity of PAT in both aqueous solution and apple juice, and evaluated for being an effective detoxifying agent for PAT. The physical and chemical properties of adsorbents were characterized by X-ray Diffraction (XRD), Fourier Transform Infrared Spectroscopy (FT-IR) and Scanning Electron Microscopy (SEM). The maximum adsorption capacities Fe3O4-COOH and Fe3O4-NH2 for PAT obtained at 25 °C, pH 6 for 5 h were 0.206 mg/g and 0.104 mg/g, respectively. Best fitting adsorption isotherm and kinetics models of PAT adsorption process were Hill isotherm (Radj2 = 0.985) and pseudo-second kinetic models (Radj2 > 0.99) for carboxy groups and Langmuir isotherm (Radj2 = 0.974) and pseudo-second kinetic models (Radj2 > 0.99) for amine groups on the surface of adsorbents. These models with the experimental results confirmed the physical adsorption process, while thermodynamic analysis indicated that adsorption process was spontaneous and endothermic between PAT and both adsorbents. Reusability study showed the effective removal of PAT at four cycles by both adsorbents. This study indicated that carboxy- and amine-terminated magnetic spheres have promising potentials in PAT removal from both aqueous solution and apple juice without affecting quality parameters of juice.

Label free detection of auramine O by G-quadruplex-based fluorescent turn-on strategy.

Auramine o (AO) is a synthetic dye used in paper and textile industries. Although it has been an unauthorized food additive in many countries due to its toxic and carcinogenic possibility, its illegal uses have been detected in certain food products such as pasta, semolina and spices and also in pharmaceuticals. The presence of AO in food products should be monitored, therefore, to minimize the negative health effects on consumers. In this study, a simple, highly sensitive and selective label free detection method was investigated for AO by G-quadruplex-based fluorescent turn-on strategy. The optimum fluorescent detection assay was achieved with a specific G-quadruplex DNA sequence, c-myc, at 400 nM in Tris-HCl buffer at pH 7.4. The linearity of fluorescence intensity depending on AO concentration ranged from 0 to 0.07 µM and LOD and LOQ were 3 nM and 10 nM, respectively. The G-quadruplex-based detection assay was highly specific for AO as compared to other two synthetic food colorings and successfully applied to determine AO in pasta, bulgur and curry powder with recoveries in the range from 70.33% to 106.49%. This G-quadruplex-based label free detection assay has a significant potential to be used in the detection of AO in food products.

Inhibition of cell migration and invasion by ICAM-1 binding DNA aptamers.

Cancer is the second leading cause of death worldwide and most of the cancer-related deaths result from metastasis. As expressed on the surface of various cancer cell types, intercellular adhesion molecule-1 (ICAM-1) has been shown to play a role in the attachment, invasion and migration of tumor cells. In this study, DNA aptamers were generated against ICAM-1 by cell-SELEX and protein SELEX method using ICAM-1(+) CHO-ICAM-1 cells and ICAM-1 protein, respectively. The pools obtained at the end of the 10th round of both SELEX were sequenced and the most enriched sequences were characterized for their binding behaviors and affinities to ICAM-1(+) CHO-ICAM-1 and ICAM-1(-) MIA PaCa-2 cells. Moreover, the inhibition abilities of sequences on migration and invasion were measured. The seven aptamer sequences were obtained selectively binding to CHO-ICAM-1 cells with Kd values in the ranging from 13.8 to 47.1 nM. Four of these aptamers showed inhibition in both migration and invasion of CHO-ICAM-1 cells at least 61%. All these results suggested that these aptamers have potential to detect specifically ICAM-1 expressing tumor cells and inhibit migration and invasion by blocking ICAM-1 related interactions of circulating tumor cells.

Adsorptive removal of patulin from apple juice via sulfhydryl-terminated magnetic bead-based separation.

Patulin is a naturally produced toxin having potential carcinogenic properties. It is mainly produced by species of Penicillium growing on fruits, especially on apples. Detoxification of patulin, therefore, is very important for providing food safety. In this study, sulfhydryl-terminated, silica coated iron oxide beads were investigated as a multi-use sorbents for effective recovery of patulin from apple juice. The effect of beads amount, contact time, pH and temperature were evaluated for high adsorption capacity and the results showed maximum adsorption capacity at a reaction condition of 1.5 mg of beads for 4 h incubation at pH 7.2 and 25 °C. These sorbents were highly effective both in aqueous solution and apple juice with adsorption efficiencies of 99% and 71.25%, respectively. Adsorption process was explained by Langmuir isotherm model with pseudo-second order kinetic model. Thermodynamic parameters described spontaneous adsorption of patulin onto beads with high feasibility and preferences. The reusability of sulfhydryl coated magnetic beads was shown for at least four times without any significant decreases in efficiency. Results showed that this sorbent had potential for removal of patulin from apple juice without any negative effects on final quality parameters, Brix, color, clarity, total sugar and titratable acidity.

DNA aptamer-based colorimetric detection platform for Salmonella Enteritidis.

Food safety is a major issue to protect public health and a key challenge is to find detection methods for identification of hazards in food. Food borne infections affects millions of people each year and among pathogens, Salmonella Enteritidis is most widely found bacteria causing food borne diseases. Therefore, simple, rapid, and specific detection methods are needed for food safety. In this study, we demonstrated the selection of DNA aptamers with high affinity and specificity against S. Enteritidis via Cell Systematic Evolution of Ligands by Exponential Enrichment (Cell-SELEX) and development of sandwich type aptamer-based colorimetric platforms for its detection. Two highly specific aptamers, crn-1 and crn-2, were developed through 12 rounds of selection with Kd of 0.971µM and 0.309µM, respectively. Both aptamers were used to construct sandwich type capillary detection platforms. With the detection limit of 103 CFU/mL, crn-1 and crn-2 based platforms detected target bacteria specifically based on color change. This platform is also suitable for detection of S. Enteritidis in complex food matrix. Thus, this is the first to demonstrate use of Salmonella aptamers for development of the colorimetric aptamer-based detection platform in its identification and detection with naked eye in point-of-care.

Evaluation of Staphylococcus aureus DNA aptamer by enzyme-linked aptamer assay and isothermal titration calorimetry.

To monitor the specificity of Staphylococcus aureus aptamer (SA-31) against its target cell, we used enzyme-linked aptamer assay. In the presence of target cell, horseradish peroxidase-conjugated streptavidin bound to biotin-labeled SA-31 showed specific binding to S  aureus among 3 different bacteria with limit of detection of 103 colony-forming unit per milliliter. The apparent Ka was 1.39 µM-1  ± 0.3 µM-1 . The binding of SA-31 to membrane proteins extracted from cell surface was characterized using isothermal titration calorimetry, and the effect of changes in binding temperature and salt concentrations of binding buffer was evaluated based on thermodynamic parameters (Ka , ΔH, and ΔG). Since binding of aptamer to its targets solely depends on its 3-dimensional structure under experimental conditions used in selection process, the change in temperature and ion concentration changed the affinity of SA-31 to its target on surface of bacteria. At 4°C, SA-31 did not show an affinity to its target with poor heat change upon injection of membrane fraction to aptamer solution. However, the apparent association constants of SA-31 slightly varied from Ka  = 1.56 µM-1  ± 0.69 µM-1 at 25°C to Ka  = 1.03 µM-1  ± 0.9 µM-1 at 37°C. At spontaneously occurring exothermic binding reactions, affinities of S aureus aptamer to its target were also 9.44 µM-1  ± 0.38 µM-1 at 50mM, 1.60 µM-1  ± 0.11 µM-1 at 137mM, and 3.28 µM-1  ± 0.46 µM-1 at 200 mM of salt concentration. In this study, it was demonstrated that enzyme-linked aptamer assay and isothermal titration calorimetry were useful tools for studying the fundamental binding mechanism between a DNA aptamer and its target on the outer surface of S aureus.

In vitro Selection of DNA Aptamers to Glioblastoma Multiforme.

Aptamer probes for specific recognition of glioblastoma multiforme were generated using a repetitive and broad cell-SELEX-based procedure without negative selection. The 454 sequencing technology was used to monitor SELEX, and bioinformatics tools were used to identify aptamers from high throughput data. A group of aptamers were generated that can bind to target cells specifically with dissociation constants (K(d)) in the nanomolar range. Selected aptamers showed high affinity to different types of glioblastoma cell lines, while showing little or no affinity to other cancer cell lines. The aptamers generated in this study have potential use in different applications, such as probes for diagnosis and devices for targeted drug delivery, as well as tools for molecular marker discovery for glioblastomas.