Sialic Acid-Functionalized Gold Nanoparticles for Sensitive and Selective Colorimetric Determination of Serotonin.

We present a novel colorimetric method inspired by nature's complex mechanisms, capable of selectively determining serotonin with high sensitivity. This method exploits the inherent binding affinity of serotonin with sialic acid (SA) molecules anchored to gold nanoparticles (SA-AuNPs). Upon serotonin binding, SA-AuNPs aggregate, and a characteristic red shift in the absorbance of SA-AuNPs accompanied by a dramatic color change (red to blue) occurs, readily observable even without instrumentation. The proposed method effectively eliminates interventions from potential interfering species such as dopamine, epinephrine, l-tyrosine, glucosamine, galactose, mannose, and oxalic acid. The absence of a color change with l-tryptophan, a structurally related precursor of serotonin, further confirms the high selectivity of this approach for serotonin detection. The colorimetric method has a wide linear dynamic range (0.05-1.0 µM), low limit of detection (0.02 µM), and fast response time (5 min). The limit of detection of the method is lower than other colorimetric serotonin sensors reported so far. The possible use of the proposed method in biological sample analysis was evaluated by employing a serotonin recovery assay in processed human plasma. The recoveries ranged from 90.5 to 104.2%, showing promising potential for clinical applications.

The potential application of gold-apoferritin nanocages conjugated with 2-amino-2-deoxy-glucose for imaging of breast cancer cells.

Development of biocompatible and multifunctional nanoprobes for tumor targeting, imaging, and therapy still remains a great challenge. Herein, gold nanoparticles (AuNPs) were synthesized in the cavity of horse spleen apoferritin protein (HoSAF) and protein surface was labeled with 2-amino-2-deoxy-glucose (2DG) as a cell surface glucose transport protein specific targeting probe to study the feasibility of its usage as a computer tomography (CT) contrast agent with tumor targeting capability through in vitro experiments. 2DG conjugated and gold-loaded apoferritin (Au-HoSAF-2DG) nanoparticles (NPs) showed selective targeting for human breast adenocarcinoma (MCF-7) cells when compared to normal breast (MCF-10A) cells. This AuNP-based imaging agent was found to be non-cytotoxic in a given concentration range with an apoptotic effect upon longer exposure times towards MCF-7 cells, while MCF-10A cells were affected less. This selective cell death would also be useful for further cancer treatments with the ability of X-ray attenuation in in vitro X-ray and computed tomography (CT) imaging.

Development of a solid-phase microextraction LC-MS/MS method for determination of oxidative stress biomarkers in biofluids.

Recently the connection between oxidative stress and various diseases, including cancer and Alzheimer's, attracts notice as a pathway suitable for diagnostic purposes. 8-Oxo-deoxyguanosine and 8-oxo-deoxyadenosine produced from the interaction of reactive oxygen species with DNA become prominent as biomarkers. Several methods have been developed for their determination in biofluids, including solid-phase extraction and enzyme-linked immunosorbent assays. However, still, there is a need for reliable and fast analytical methods. In this context, solid-phase microextraction offers many advantages such as flexibility in geometry and applicable sample volume, as well as high adaptability to high-throughput sampling. In this study, a solid-phase microextraction method was developed for the determination of 8-oxo-deoxyguanosine and 8-oxo-deoxyadenosine in biofluids. The extractive phase of solid-phase microextraction consisted of hydrophilic-lipophilic balanced polymeric particles. In order to develop a solid-phase microextraction method suitable for the determination of the analytes in saliva and urine, several parameters, including desorption solvent, desorption time, sample pH, and ionic strength, were scrutinized. Analytical figures of merit indicated that the developed method provides reasonable interday and intraday precisions (<15% in both biofluids) with acceptable accuracy. The method provides a limit of quantification for both biomarkers at 5.0 and 10.0 ng/mL levels in saliva and urine matrices, respectively.

EF2-kinase targeted cobalt-ferrite siRNA-nanotherapy suppresses BRCA1-mutated breast cancer.

Aim: To investigate the role of EF2K in BRCA1-mutated breast cancer. Materials & methods: We developed silica coated cobalt-ferrite (CoFe) nanoparticles for in vivo delivery of small interfering RNAs (siRNAs) into BRCA1-mutated breast cancer. Results: Expression of EF2K is highly upregulated in the majority (78.5%) of BRCA1-mutated patients and significantly associated with poor patient survival and metastasis. Silencing of EF2K reduced cell proliferation, migration and invasion of the cancer cells. In vivo therapeutic targeting of EF2K by CoFe-siRNA-nanoparticles leads to sustained EF2K gene knockdown and suppressed tumor growth in orthotopic xenograft models of BRCA1-mutated breast cancer. Conclusion: EF2K is a potential novel molecular target in BRCA1-mutated tumors and CoFe-based siRNA nanotherapy may be used as a novel approach to target EF2K.

Cellular uptake and apoptotic potential of rhenium labeled magnetic protein cages in MDA-MB-231 cells.

188Re-magnetoferritin nanoparticles (NPs) provide an attractive platform for localized radiation therapy due to their magnetic targeting capability while enhancing contrast in magnetic resonans imaging (MRI) signals. In this study, cellular uptake, in vitro cytotoxicity, apoptotic potential of a non-radioactive isotope of rhenium in the form of 187Re-magnetoferritin NPs were evaluated in both human normal mammary epithelial and breast metastatic adenocarcinoma cell lines. The results showed that, NP administration into the cells is through receptor mediated endocytosis and cancer cells displayed significantly higher uptake and cytotoxicity compared to normal cells. IC50 values of nanoparticles were calculated as 0.96 mg/mL for cancer and 1.73 mg/mL for normal cells. Annexin V/ Propidium Iodide (PI) staining also showed that, NPs induced higher apoptotic rates in cancer cells compared to normal cells. Gene expression analyses confirming the results showed that, pro-apoptotic PUMA and BAX genes were significantly up-regulated while anti-apoptotic BCL-2 and SURVIVIN genes were down-regulated in cancer cells compared to normal cells. Overall, these in vitro results suggest that, 187Re-magnetoferritin NPs have a promising potential for cancer therapy and can be used for imaging and diagnostic purposes for breast cancer at concentrations lower than 0.96 mg/mL. At concentrations above 1 mg/mL, NPs induce apoptosis which can also be used for cancer treatments.

A Novel Acetylcholinesterase Biosensor: Core-Shell Magnetic Nanoparticles Incorporating a Conjugated Polymer for the Detection of Organophosphorus Pesticides.

To construct a sensing interface, in the present work, a conjugated polymer and core-shell magnetic nanoparticle containing biosensor was constructed for the pesticide analysis. The monomer 4,7-di(furan-2-yl)benzo[c][1,2,5]thiadiazole (FBThF) and core-shell magnetic nanoparticles were designed and synthesized for fabrication of the biosensing device. The magnetic nanoparticles were first treated with silica and then modified using carboxyl groups, which enabled binding of the biomolecules covalently. For the construction of the proposed sensor a two-step procedure was performed. First, the poly(FBThF) was electrochemically generated on the electrode surface. Then, carboxyl group modified magnetic nanoparticles (f-MNPs) and acetylcholinesterase (AChE), the model enzyme, were co-immobilized on the polymer-coated surface. Thereby, a robust and novel surface, conjugated polymer bearing magnetic nanoparticles with pendant carboxyl groups, was constructed, which was characterized using Fourier transform infrared spectrometer, cyclic voltammetry, scanning electron microscopy, and contact angle measurements. This novel architecture was then applied as an immobilization platform to detect pesticides. To the best of our knowledge, a sensor design that combines both conjugated polymer and magnetic nanoparticles was attempted for the first time, and this approach resulted in improved biosensor characteristics. Hence, this approach opens a new perspective in the field of enzyme immobilization and sensing applications. Paraoxon and trichlorfon were selected as the model toxicants. To obtain best biosensor performance, optimization studies were performed. Under optimized conditions, the biosensor in concern revealed a rapid response (5 s), a low detection limit (6.66 × 10(-3) mM), and high sensitivity (45.01 µA mM(-1) cm(-2)). The KM(app) value of poly(FBThF)/f-MNPs/AChE were determined as 0.73 mM. Furthermore, there was no considerable activity loss for 10 d for poly(FBThF)/f-MNPs/AChE biofilm.

2-Amino-2-deoxy-glucose conjugated cobalt ferrite magnetic nanoparticle (2DG-MNP) as a targeting agent for breast cancer cells.

In this study, 2-amino-2-deoxy-glucose (2DG) was conjugated to COOH modified cobalt ferrite magnetic nanoparticles (COOH-MNPs), which were designed to target tumor cells as a potential targetable drug/gene delivery agent for cancer treatment. According to our results, it is apparent that, 2DG labeled MNPs were internalized more efficiently than COOH-MNPs under the same conditions in all cell types (MDA-MB-231 and MCF-7 cancer and MCF-10A normal breast cells) (p<0.001). Moreover, the highest amount of uptake was observed in MDA-MB-231, followed by MCF-7 and normal MCF-10A cells for both MNPs. The apoptotic effects of 2DG-MNPs were further evaluated, and it was found that apoptosis was not induced at low concentrations of 2DG-MNPs in all cell types, whereas dramatic cell death was observed at higher concentrations. In addition, the gene expression levels of four drug-metabolizing enzymes, two Phase I (CYP1A1, CYP1B1) and two Phase II (GSTM3, GSTZ1) were also increased with the high concentrations of 2DG-MNPs.

Cellular uptake, genotoxicity and cytotoxicity of cobalt ferrite magnetic nanoparticles in human breast cells.

Magnetic nanoparticles (MNPs) have been increasingly used for many years as MRI agents and for gene delivery and hyperthermia therapy, although there have been conflicting results on their safety. In this study, cobalt ferrite magnetic nanoparticles (CoFe-MNPs) were prepared by the co-precipitation method and their surfaces were modified with silica by the sol-gel method. The particle and hydrodynamic sizes, morphology and crystal structure of the bare and silica-coated CoFe-MNPs were evaluated by transmission electron microscopy (TEM), dynamic light scattering (DLS), X-ray diffraction spectroscopy (XRD) and Fourier transform infrared spectroscopy (FTIR). The size of the bare CoFe-MNPs was in the range 8-20 nm and they were homogeneously coated with 3-4 nm silica shells. The bare and silica-coated CoFe-MNPs were agglomerated at physiological pH. However, the sizes of the agglomerates were below 200 nm both in water and complete medium. The cytotoxic and genotoxic potentials of the bare and silica-coated CoFe-MNPs were evaluated in a metastatic breast cancer cell line, MDA-MB-231, as well as a noncancerous mammary epithelial cell line, MCF-10A, by using XTT cytotoxicity, single-cell gel electrophoresis (comet), and cytokinesis-blocked (CB) micronucleus (CBMN) assays. Characterization studies with TEM, inductively coupled plasma optical emission spectroscopy (ICP-OES) and Prussian blue staining indicated that the CoFe-MNPs were internalized into the cells by energy-dependent endocytosis. The highest amount of uptake was observed in the cancer cells and the uptake of the silica-coated CoFe-MNPs was higher than that of the bare ones in both cell lines. The bare CoFe-MNPs showed higher levels of both cytotoxicity and genotoxicity than the silica-coated CoFe-MNPs. Moreover, the cancer cells seemed to be more susceptible to the CoFe-MNPs' toxicity compared to the noncancerous cells. There was a concentration and time-dependent increase in DNA damage and the micronucleus (MN) frequency, which was statistically significant starting with the lowest concentration of bare CoFe-MNPs (p < 0.05), while no significance was observed below the concentration of 250 µg mL-1 for the silica-coated MNPs. Also, the extent of both DNA damage and MN frequency was much higher in the cancer cells compared to the noncancerous cells. According to our results, the silica coating ameliorated both the cytotoxicity and genotoxicity as well the internalization of the CoFe-MNPs.

Copper(0) nanoparticles supported on silica-coated cobalt ferrite magnetic particles: cost effective catalyst in the hydrolysis of ammonia-borane with an exceptional reusability performance.

Herein we report the development of a new and cost-effective nanocomposite catalyst for the hydrolysis of ammonia-borane (NH(3)BH(3)), which is considered to be one of the most promising solid hydrogen carriers because of its high gravimetric hydrogen storage capacity (19.6% wt) and low molecular weight. The new catalyst system consisting of copper nanoparticles supported on magnetic SiO(2)/CoFe(2)O(4) particles was reproducibly prepared by wet-impregnation of Cu(II) ions on SiO(2)/CoFe(2)O(4) followed by in situ reduction of the Cu(II) ions on the surface of magnetic support during the hydrolysis of NH(3)BH(3) and characterized by ICP-MS, XRD, XPS, TEM, HR-TEM and N(2) adsorption-desorption technique. Copper nanoparticles supported on silica coated cobalt(II) ferrite SiO(2)/CoFe(2)O(4) (CuNPs@SCF) act as highly active catalyst in the hydrolysis of ammonia-borane, providing an initial turnover frequency of TOF = 2400 h(-1) at room temperature, which is not only higher than all the non-noble metal catalysts but also higher than the majority of the noble metal based homogeneous and heterogeneous catalysts employed in the same reaction. More importantly, they were easily recovered by using a permanent magnet in the reactor wall and reused for up to 10 recycles without losing their inherent catalytic activity significantly, which demonstrates the exceptional reusability of the CuNPs@SCF catalyst.

New approach for the surface enhanced resonance Raman scattering (SERRS) detection of dopamine at picomolar (pM) levels in the presence of ascorbic acid.

The development of a novel surface-enhanced resonance Raman scattering (SERRS) platform that allows fast and sensitive detection of dopamine (DA) has been reported. The iron-nitrilotriacetic acid attached silver nanoparticle (Ag-Fe(NTA)) substrate provides remarkable sensitivity and reliable repeatability. The advantages of both the surface functionalization for specific analytes and the SERRS are integrated into a single functional unit. While the silver core gives the necessary enhancing properties, the Fe-NTA receptors can trap DA adjacent the silver core and the NTA-Fe-DA complex formed provides resonance enhancement with a 632.8 nm laser. DA could be detected in pM level without any pretreatment with a reliable discrimination against AA, by utilizing low laser power (10 mW) and short data acquisition time (10 s). The high sensitivity along with the improved selectivity of this sensing approach is a significant step toward molecular diagnosis of Parkinson's disease.

Germanium determination by flame atomic absorption spectrometry: an increased vapor pressure-chloride generation system.

A new chloride generation system was designed for the direct, sensitive, rapid and accurate determination of the total germanium in complex matrices. It was aimed to improve the detection limit of chloride generation technique by increasing the vapor pressure of germanium tetrachloride (GeCl(4)). In order to do so, a novel joint vapor production and gas-liquid separation unit equipped with a home-made oven was incorporated to an ordinary nitrous oxide-acetylene flame atomic absorption spectrometer. Several variables such as reaction time, temperature and acid concentration have been investigated. The linear range for germanium determination was 0.1-10 ng mL(-1) for 1 mL sampling volume with a detection limit (3s) of 0.01 ng mL(-1). The relative standard deviation (RSD) was 2.4% for nine replicates of a 1 ng mL(-1) germanium solution. The method was validated by the analysis of one non-certified and two certified geochemical reference materials, respectively, CRM GSJ-JR-2 (Rhyolite), and GSJ-JR-1 (Rhyolite), and GBW 07107 (Chinese Rock). Selectivity of the method was investigated for Cd(2+), Co(2+), Cu(2+), Fe(3+), Ga(3+), Hg(2+), Ni(2+), Pb(2+), Sn(2+), and Zn(2+) ions and ionic species of As(III), Sb(III), Te(IV), and Se(IV).

Carboligation reactions with benzaldehyde lyase immobilized on superparamagnetic solid support.

Histidine-tagged recombinant benzaldehyde lyase (BAL, EC 4.1.2.38) was efficiently immobilized to surface-modified magnetic particles with affinity ligand binding. In addition to conventional benzoin condensation reactions, two important representative BAL-catalyzed carboligation reactions, were also performed with this magnetically responsive biocatalyst. The results obtained from the carboligation reactions that were performed with this simple and convenient heterogenous biocatalyst were comparable to that of free-enzyme-catalyzed reactions.

Preparation and characterization of Ni-nitrilotriacetic acid bearing poly(methacrylic acid) coated superparamagnetic magnetite nanoparticles.

Stable superparamagnetic magnetite (Fe3O4) nanoparticles were synthesized via co-precipitation in the presence of poly(methacrylic acid) (PMAA) in aqueous solution. The polymer coated Fe3O4 nanoparticles were characterized using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction, thermal analysis, and vibrating sample magnetometry (VSM) techniques. These measurements reveal the presence of magnetite nanoparticles with a size of approximately 8 nm inside the PMAA matrix. The magnetization value of these superparamagnetic nanoparticles at room temperarure and 7 T was measured as about 40 emu/g. PMAA-coated Fe3O4 nanoparticles were further assembled with Ni-chelate through a reaction between a primary amine-bearing NTA (nitrilotriacetic acid) ligand and carboxy-functional groups of PMAA. NTA-PMAA-coated magnetite nanoparticles were then loaded with nickel ions and characterized using FTIR. The average amount of binded Ni on the surface of the NTA-modified PMAA coated Fe3O4 was calculated as 1.65 +/- 0.3 x 10(-6) mol nickel(II) ions per g of the magnetic particles from the inductively coupled plasma optical emission spectroscopy (ICP-OES) measurements.

Rapid synthesis and characterization of maghemite nanoparticles.

Fe2O3-SiO2 nanocomposites were prepared by a sol-gel method using various evaporation surface to volume (S/V) ratios ranging from 0.03 to 0.2. The Fe2O3-SiO2 sols were gelated at various temperatures ranging from 50 degrees C to 70 degrees C, and subsequently they were calcined in air at 400 degrees C for 4 hours. The structure and the magnetic properties of the prepared Fe203-SiO2 nanocomposites were characterized using X-ray diffraction (XRD), transmission electron microscopy (TEM), differential thermal analysis (DTA), and vibrating sample magnetometer (VSM) measurements. The gelation temperature of the Fe2O3-SiO2 sols influenced strongly the particle size and crystallinity of the maghemite nanoparticles. It was observed that the particle size of maghemite nanoparticles increased with the increasing of the gelation temperature of the sols, which may be due to the agglomeration of the maghemite particles at elevated temperatures inside the microporosity of the silica matrix during the gelation process, and the subsequent calcination of these gels at 400 degrees C resulted in the formation of large size iron oxide particles. Magnetization studies at temperatures of 10, 195, and 300 K showed superparamagnetic behavior for all the nanocomposites prepared using the evaporation surface to volume ratio (S/V) of 0.1, 0.2, 0.09, and 0.08. The saturation magnetization, Ms, values measured at 10 K were 5.5, 8.5, and 9.5 emu/g, for the samples gelated at 50, 60, and 70 degrees C, respectively. At the gelation temperature of 70 degrees C, gamma-Fe2O3 crystalline superparamagnetic nanoparticles with the particle size of 9 +/- 2 nm were formed in 12 hours for the samples prepared at the S/V ratio of 0.2.

Silver nanoparticle-doped polyvinyl alcohol coating as a medium for surface-enhanced Raman scattering analysis.

A simple polymer substrate for inducing Surface-Enhanced Raman Scattering (SERS) has been investigated. This SERS substrate consists of a solid support, such as a glass slide covered with polyvinyl alcohol (PVA) impregnated with fine silver nanoparticles. The preparation simply involves mixing aqueous PVA polymer with solid AgNO3 to produce a solution that can be easily spin coated on the glass substrate and dried to obtain a hard translucent coating. Aqueous solution of FeSO4.7H2O was used to reduce Ag+ ions to silver nanoparticles. The effects of various experimental conditions of sample preparation were investigated in order to improve the Raman enhancement efficiency of the substrate. The overall substrate performance was evaluated with the use of biologically important compounds: benzoic acid, p-amino benzoic acid, pyridine and dopamine. The spectral features of these compounds closely matched with those reported in literature. The use of the polymer matrix made the SERS substrate resistant to scratching, therefore, improving it to be more suitable for field applications. The hydrophilic nature of the polymer provides additional advantages for probing biological samples. The shelf-life of the dried, unreduced substrates is at least one month.

Selective pre-concentration of selenite from aqueous samples using mercapto-silica.

Silica gel modified with 3-mercaptopropyl-trimethoxysilane was used for the selective separation and pre-concentration of selenite (Se(IV)) from aqueous solutions containing Se(IV) and selenate (Se(VI)). Over a wide range of acidity, from 2 mol l(-1) HCl to pH 9.00, Se(IV) was taken up by the mercaptopropyl-silica with nearly 100% efficiency; Se(VI) however was unretained. Se(IV) content was determined by hydride generation atomic absorption spectrometry (HGAAS), following batch release of the selenium from the pre-concentration medium by acidic periodate. The overall pre-concentration efficiency, including both take-up and elution, in the range of 89-106%. The method was applied to spiked seawater samples containing as low as 800 ng l(-1) Se in selenite form. This solid-phase extraction system offers several major advantages over conventional solvent extraction procedures. It firstly exhibits high selectivity for Se(IV) over Se(VI). Using the solid-phase media, pre-concentration of Se(IV) in dilute water samples can be carried out in the field, stabilizing the selenite-selenium in a convenient form for transport and storage. In addition, selenium stored on silica is derived solely from Se(IV) overcoming problems of selenium redox speciation changes and loss during storage.