A Comparative Review of Key Isothiocyanates and Their Health Benefits.

Isothiocyanates are biologically active products resulting from the hydrolysis of glucosinolates predominantly present in cruciferous vegetables belonging to the Brassicaceae family. Numerous studies have demonstrated the diverse bioactivities of various isothiocyanates, encompassing anticarcinogenic, anti-inflammatory, and antioxidative properties. Nature harbors distinct isothiocyanate precursors, glucosinolates such as glucoraphanin and gluconastrin, each characterized by unique structures, physical properties, and pharmacological potentials. This comprehensive review aims to consolidate the current understanding of Moringa isothiocyanates, mainly 4-[(α-L-rhamnosyloxy) benzyl] isothiocyanate), comparing this compound with other well-studied isothiocyanates such as sulforaphane and phenyl ethyl isothiocyanates. The focus is directed toward elucidating differences and similarities in the efficacy of these compounds as agents with anticancer, anti-inflammatory, and antioxidative properties.

Immunosensors for quantifying cyclooxygenase 2 pain biomarkers.

BACKGROUND: Cyclooxygenase 2 (COX-2) is a key enzyme in pain biomarkers, inflammation and cancer cell proliferation. Thus biosensors that can quantify pain mediators based on biochemical mechanism are imperative. METHODS: Biomolecular recognition and affinity of antigenic COX-2 with the antibody were investigated using surface plasmon resonance (SPR) and ultra-sensitive portable capillary (UPAC) fluorescence sensors. Polyclonal goat anti-COX-2 (human) antibodies were covalently immobilized on gold SPR surface and direct recognition for the COX-2 antigen assessed. The UPAC sensor utilized an indirect sandwich design involving covalently attached goat anti-COX-2 as the capture antibody and rabbit anti-COX-2 (human) antibody as the secondary antibody. RESULTS: UPAC fluorescence signals were directly proportional to COX-2 at a linear range of 7.46×10⁻4-7.46×10¹ ng/ml with detection limit of 1.02×10⁻4 ng/ml. With SPR a linear range was 3.64×10⁻4-3.64×10² ng/ml was recorded and a detection limit of 1.35×10⁻4 ng/ml. Validation was achieved in simulated blood samples with percent recoveries of 81.39% and 87.23% for SPR and UPAC respectively. CONCLUSION: The developed sensors have the potential to provide objective characterization of pain biomarkers for clinical diagnoses.

Synthesis and antiproliferative activity of new coumarin-based benzopyranone derivatives against human tumor cell lines.

The synthesis and antiproliferative activity of new coumarin-based benzopyranone derivatives containing basic amino side chain are described. The cytotoxicities against A549 and MCF-7 human cancerous cell lines were determined after 24, 48, 72 h drug exposure employing MTT assay at concentrations ranging from 0-100 µM. The antiproliferative activities of these compounds were compared to tamoxifen (TAM), 4-hydroxytamoxifen (4-OHT), raloxifene (RAL), 17ß-estradiol (E2) and Diethylstilbestrol (DES). In vitro results indicated that compounds 10 and 12 were the most potent showing significant inhibitory activities against these cell lines. Furthermore, their antiproliferative activity against MCF-7 human breast cancer cell line is comparable to that of TAM, RAL and 4-OHT.

Spectroelectrochemical characterization of pain biomarkers.

This article reports the first electrochemical characterization of pain biomarkers that include arachidonic acid (AA), prostaglandin G(2) (PGG(2)), and cyclooxygenase 2 (COX-2). These biomarkers are mediators of pathophysiology of pain, inflammation, and cell proliferation in cancer. The article also reports the development of an electrochemical immunosensor for monitoring these pain biomarkers. The results revealed that direct electron transfer between AA metabolites and the electrode could be easily monitored and that an enzyme-modified electrode dramatically enhanced bioelectrocatalytic activity toward AA. Cyclic voltammetric analysis of AA revealed a concentration-dependent anodic current with a slope of 2.37 and a limit of detection (LOD) of 0.25nM. This unique AA/gold electrode electron transfer provides a good electrochemical sensing platform for prostaglandin H(2) (PGH(2)) as the basis for quantitation of pain. An amperometric signal intensity of a COX-2 antibody-modified gold electrode was linear with COX-2 concentration in the range of 0.1-0.5microg/ml and an LOD of 0.095microg/ml. The results also revealed a linear correlation of the concentration of PGG(2) with an LOD of 0.227microM.

Identification and quantitation of Bacillus globigii using metal enhanced electrochemical detection and capillary biosensor.

Presented herein are two detection strategies for the identification and quantification of Bacillus globigii, a spore forming nonpathogenic simulant of Bacillus anthracis. The first strategy involves a label-free, metal-enhanced electrochemical immunosensor for the quantitative detection of Bacillus globigii (atrophaeus). The immunosensor comprises of antibacillus globigii (BG) antibody self-assembled onto a gold quartz crystal electrode via cystamine bond. A solid-phase monolayer of silver underpotentially deposited onto the cystamine modified-Au-electrode surface is used as the redox probe. The monolayer was also generated by adsorbing silver nanoparticles on the gold electrode. When the antibody-modified electrode is exposed to BG spores, the antibody-antigen (Ab-Ag) complex formed insulated the electrode surface toward the silver redox probe. The variation of redox current was found to be proportional to the concentration of the BG spores between 1 x 10(2)-3.5 x 10(4) spores/mL. A detection limit of 602 spores/mL was obtained, which is well-below the infectious dose of anthrax spores at 2.5 x 10(5) spores/mL. The second approach involves the use of ultrasensitive portable capillary biosensor (UPAC) to detect the spores. The capillary is an enclosed system that acts as the flow cell, the waveguide, and the solid support for immobilized bimolecular probes. An evanescent excitation generates a signal from an antigen-antibody-fluorophore complex, which propagates along the capillary and is guided to the detector. A limit of detection of 112 spores/mL was reported using the UPAC sensor. Both methods showed lower detection limits compared to the conventional ELISA. The effect of potential interferants tested using Bacillus pumilus confirmed the selectivity for the analyte. This work should allow the first responders to rapidly detect and quantify Bacillus globigii spores at concentrations that are well-below the infectious dose.

Status of biomolecular recognition using electrochemical techniques.

The use of nanoscale materials (e.g., nanoparticles, nanowires, and nanorods) for electrochemical biosensing has seen explosive growth in recent years following the discovery of carbon nanotubes by Sumio Ijima in 1991. Although the resulting label-free sensors could potentially simplify the molecular recognition process, there are several important hurdles to be overcome. These include issues of validating the biosensor on statistically large population of real samples rather than the commonly reported relatively short synthetic oligonucleotides, pristine laboratory standards or bioreagents; multiplexing the sensors to accommodate high-throughput, multianalyte detection as well as application in complex clinical and environmental samples. This article reviews the status of biomolecular recognition using electrochemical detection by analyzing the trends, limitations, challenges and commercial devices in the field of electrochemical biosensors. It provides a survey of recent advances in electrochemical biosensors including integrated microelectrode arrays with microfluidic technologies, commercial multiplex electrochemical biosensors, aptamer-based sensors, and metal-enhanced electrochemical detection (MED), with limits of detection in the attomole range. Novel applications are also reviewed for cancer monitoring, detection of food pathogens, as well as recent advances in electrochemical glucose biosensors.

Tuning the surfaces of palladium nanoparticles for the catalytic conversion of Cr(VI) to Cr(III).

This work reports the feasibility of using Pd nanoparticles as innovative catalysts in the conversion of reducible contaminants from toxic to benign forms. Cr(VI) is a known carcinogen while the trivalent chromium salts are believed to be non-toxic. The ability of Pd nanoparticles to catalyze the rapid reduction of Cr(VI) to Cr(III) using reactive sulfur intermediates produced in situ was therefore studied. Using a microchamber set at 130 degrees C, the reduction mixture consists of palladium nanoparticles and sulfur (PdNPs/S), which generated highly reducing sulfur intermediates that effected the reduction of Cr(VI) to Cr(III) by 1st order reaction kinetics. UV-visible spectroscopy and cyclic voltammetry were employed to monitor the reduction process. The results showed that 99.8% of 400 microM Cr(VI) was reduced to Cr(III) by PdNPs/S in one hour compared to 2.1% by a control experiment consisting of sulfur only. The rate of Cr(VI) reduction was found to be dependent on temperature and pH and was greatly enhanced by the addition of PdNPs. Subsequent application of this approach in the reduction of Cr(VI) in soil and aqueous media was conducted. In contrast to the control experiments with and without PdNPs or sulfur, greater than 92% conversion rate was obtained in the presence of PdNPs/S within 1 hour. This represents over a 500-fold improvement in conversion rate compared to current microbial approaches. XPS analysis provided the confirmation regarding the oxidation states of Cr(VI), Cr(III) and the nature of the reactive intermediates. This work offers PdNPs/S as a new interface for the reduction of high oxidation state heavy metal pollutants.

Advanced electrochemical sensors for cell cancer monitoring.

The possibility of using minimally invasive analytical instruments to monitor cancerous cells and their interactions with analytes provide great advances in cancer research and toxicology. The real success in the development of a reliable sensor for cell monitoring depends on the ability to design powerful instrumentation that will facilitate efficient signal transduction from the biological process that occurs in the cellular environment. The resulting sensor should not affect cell viability and must function as well as adapt the system to the specific conditions imposed by the cell culture. Due to their performance, electrochemical biosensors could be used as an effective instrument in cell cancer research for studying biochemical processes, cancer development and progression as well as toxicity monitoring. Current research in this direction is conducted through high-throughput, compact, portable, and easy to use sensors that enable measurement of cells' activity in their optimum environment. This paper discusses the potential of a high-throughput electrochemical multisensor system, so-called the DOX system for monitoring cancerous cells and their interaction with chemical toxins. We describe the methodology, experiments, and the operation principle of this device, and we focus on the challenges encountered in optimizing and adapting the system to the specific cell-culture conditions. The DOX system is also compared with conventional cell-culture techniques.

Development of an oral biosensor for salivary amylase using a monodispersed silver for signal amplification.

An amperometric biosensor for monitoring the level of protein amylase in human saliva is described. A novel design and the preparation of amylase antibodies and antigens, essential for the development of the biosensor, are reported. The biosensor sensing elements comprise a layer of salivary antibody (or antigen) self-assembled onto Au-electrode via covalent attachment. Molecular recognition between the immobilized antibody and the salivary amylase proteins was monitored via an electroactive indicator (e.g., K(3)Fe(CN)(6)) or a monodispersed silver layer present in solution or electrochemically deposited onto the solid electrode. This electroactive indicator was oxidized or reduced and the resulting current change provided the analytical information about the concentration of the salivary proteins. The limit of detection of 1.57 pg/ml was obtained, in comparison to detection limits of 4.95 pg/ml obtained using potassium ferrocyanide as the redox probe and 10 ng/ml obtained using enzyme-linked immunosorbent assay. Cross-reactivity was tested against cystatin antibodies and was found to be less than 2.26%.

Effect of natural and synthetic estrogens on a549 lung cancer cells: correlation of chemical structures with cytotoxic effects.

A series of synthetic (nonylphenol, diethylstilbestrol, and bisphenol A) and natural (quercetin, resveratrol, and genistein) phenolic estrogens were investigated for their ability to affect the viability and proliferation of A549 lung cancer cells. To assess and distinguish the cytotoxic effect of individual estrogens, we used both the MTT tetrazolium spectrophotometric method and the fluorescence assay, while the induction of the cell specific apoptotic process was examined by fluorescence microscopy after treatment of cells with SYTO 24 green fluorescent dye. A systematic study of interferences for both fluorescence and MTT methods is presented. The results showed that both natural and synthetic estrogens decreased the viability and proliferation of A549 lung cancer cells in a dose-dependent manner but at different sensitivities. Nonylphenol appeared very different as compared to the other estrogens, acting by inducing the higher inactivation rate of the cells within a very short time. The cytotoxic effect of the estrogens was directly related to their structural and conformational characteristics including chain length, number, and position of hydroxyl groups and degree of saturation.

Autonomous multielectrode system for monitoring the interactions of isoflavonoids with lung cancer cells.

The cytotoxic effect of isoflavonoids in the development of different forms of cancer has been reported by epidemiological and dietary studies. Consequently, there is a search for an accurate and reliable method for monitoring the interactions of these chemicals with cancerous cells. We have developed and optimized a fully autonomous electrochemical biosensor for studying the role of isoflavonoids on A549 lung adenocarcinoma cell line. This advanced biosensor uses a prototype 96-electrode (DOX-96) well-type device that allows the measurement of cell respiratory activity via the consumption of dissolved oxygen. The system provides a continuous, real-time monitoring of cell activity upon exposure to naturally occurring polyphenols, specifically resveratrol, genistein, and quercetin. The system is equipped with a multipotentiostat, a 96-electrode well for measurements and cell culturing with 3 disposable electrodes fitted into each well. A comparison with classical "cell culture" techniques indicates that the biosensor provides real-time measurement with no added reagents. A detection limit of 1 x 10(4) was recorded versus 200 and 6 x 10(3) cells/well for MTT and fluorescence assays, respectively. This method was optimized with respect to cell stability, reproducibility, applied potential, cell density per well, volume/composition of cell culture medium per well, and incubation. Others include total measuring time, temperature, and sterilization procedure. This study represents a basic research tool that may allow researchers to study the type, level, and specific influence of isoflavonoids on cells.

Novel fluorescent biosensor for pathogenic toxins using cyclic polypeptide conjugates.

This work describes a two-step conjugate synthesis of a new fluorescent analog of microcystin-LR and its subsequent utilization for the development of an optical biosensor for cyanobacteria toxins. The biosensor concept is based on the competitive binding between the native microcystin and its fluorescent analog at immobilized alkaline phosphatase enzymes.

Correlation of analyte structures with biosensor responses using the detection of phenolic estrogens as a model.

The apparent increase in hormone-induced cancers and disorders of the reproductive tract in wildlife and humans has led to a search for an accurate and reliable method for monitoring endocrine-disrupting chemicals (EDCs). This study presents a generic approach that may allow researchers to establish screening procedures for potential EDCs by correlating the analyte structures with biosensor responses and explain possible reaction mechanisms. A simple amperometric tyrosinase-based biosensor (Tyr-CPE) has been developed for the detection of phenolic EDCs. The investigation of the enzymatic oxidation of selected phenolic estrogens was first carried out using UV-vis spectroscopy. The result was used to correlate sensor responses to enzymatic activity. Natural phytoestrogen polyphenols, including resveratrol (RES), genistein (GEN), and quercetin (QRC), were compared with synthetic estrogens, for example, bisphenol A (BPhA), nonylphenol (NPh), and diethylstilbestrol (DES). The Tyr-CPE biosensor resulted in rapid, simple, and accurate measurement of phenolic estrogens with varying degrees of sensitivity, selectivity, and response times. The sensor responses have been evaluated for the detection of binary and tertiary mixtures of EDCs and natural estrogens. The results showed that BPhA could be successfully discriminated in a composite mixture containing NPh and DES at various ratios. In the case of natural phenolic estrogens GEN, RES, and QRC, the sensor allows the determination of a total phenolic content. The sensor was also validated for the detection of BPhA in a real environmental water sample, and the results was compared with standard ASTM method 9065. Mechanistically, our results indicated that the number of OH groups, the nature and the position of aryl ring substituents, or both could affect the detection limit and the biosensor sensitivity.