Direct electrochemical determination of environmentally harmful pharmaceutical ciprofloxacin in 3D printed flow-through cell.

Rising amounts of antibiotic residues in wastewater cause serious problems including increased bacterial resistance. Wastewater treatment plants (WWTPs) do not, in the case of new, modern pharmaceuticals, ensure their complete removal. Ciprofloxacin (CIP) is one of many micropollutants that partially pass through WWTPs, implying that its monitoring is essential for the assessment of the water quality. In real sewage systems, the determination of CIP needs to be performed under flowing conditions, which calls for the deployment of inexpensive, robust, and easily integrable approaches such as electrochemical techniques. However, to the best of our knowledge, there is no report on the electrochemical determination of CIP in a flowing matrix. To bridge this gap, we perform here cyclic and square-wave voltammetric sensing study of CIP employing boron-doped diamond screen printed electrodes in a custom-made 3D printed flow-through cell to mimic conditions in real sewage systems. An irreversible two-step oxidation of CIP is demonstrated, with the first step providing clear Faradaic response as analytically relevant signal. This response was found to scale with the sample flow rate according to the prediction given by Levich equation. Our work provides an in-depth inspection of the electrochemical response of CIP under controlled-convection conditions, which is an essential prerequisite for monitoring this antibiotic in real flowing sewage systems.

Effect of Triclosan and Silver Nanoparticles on DNA Damage Investigated with DNA-Based Biosensor.

Triclosan (TCS) is a broad-spectrum antimicrobial agent widely used in personal care, healthcare, and clinical practice. One of the most important aspects of toxicological profiling of compounds is their interaction with DNA. In human cells, TCS causes a significant reduction in DNA methylation. The involvement of TCS in chromosomal aberrations, DNA damage, and strand breaks, as well as DNA damage from TCS degradation products, was reported. AgNPs share similarities with TCS in terms of antimicrobial properties, enter the body after exposure, and are used even together with TCS in oral care products. Therefore, their mutual effect on the DNA is of interest. In this study, the electrochemical behavior of TCS on a glassy carbon electrode (GCE) and the biosensor with salmon sperm dsDNA (DNA/GCE), DNA damage by TCS present in phosphate buffer solution pH 7.4 and an additional effect of the immobilized AgNP layer on such DNA damage have been investigated. Two different sizes of AgNPs (about 15 and 37 nm) were tested. Using square-wave voltammetric signals of nucleobases, the portion of survived DNA was 64% in the presence of 15 nm AgNPs compared to 55% in its absence. The protective effect of AgNPs on DNA against TCS-induced DNA damage was found.

Gallic acid-coated silver nanoparticles as perspective drug nanocarriers: bioanalytical study.

The ability of silver nanoparticles (AgNPs) to be used as drug nanocarriers has helped rapidly to invent novel strategies to treat diseases, such as cancer. The nanoparticles may offer a valuable tool to novel pH-sensitive drug delivery systems in the present scenario because of their undergoing mechanisms associated with the regulated dissolution, aggregation, and generation of oxygen radicals as well. These processes could be monitored by electrochemical (bio)sensors that are less money and time-consuming compared to other analytical approaches, however, with comparable analytical performance. In this paper, synthesized and microscopically characterized gallic acid-coated AgNPs (GA-AgNPs) are investigated using spectral and electrochemical methods. To investigate the Ag+ release, a 21-day ageing experiment is performed spectrophotometrically, finding that the peak maximum of GA-AgNPs spectra diminished by 24.5%. The highest Ag+ content was electrochemically determined in the supernatant solution after centrifugation (6.97 µmol·L-1), while no significant concentration of silver ions in solution after redispersion was observed (1.26 µmol·L-1). The interaction experiment indicates a stabilization of GA-AgNPs in the presence of long-chain dsDNA as well as a mutual electrostatic interaction with DNA sugar-phosphate backbone. This interaction mechanism is confirmed by FTIR analysis, showing a shift (1049 to 1061 cm-1 and 913 to 964 cm-1) specific to DNA phosphate bands. Finally, doxorubicin-loaded GA-AgNPs are monitored for the specific drug release in the physiological and more reactive weakly acidic microenvironment. Hereby, electrochemical (bio)sensing of GA-AgNPs undergoing mechanisms shows a huge potential to be used for monitoring of drug delivery systems at cancer therapy.

Advanced materials-integrated electrochemical sensors as promising medical diagnostics tools: A review.

Electrochemical sensors have increasingly been linked with terms as modern biomedically effective highly selective and sensitive devices, wearable and wireless technology, portable electronics, smart textiles, energy storage, communication and user-friendly operating systems. The work brings the overview of the current advanced materials and their application strategies for improving performance, miniaturization and portability of sensing devices. It provides the extensive information on recently developed (bio)sensing platforms based on voltammetric, amperometric, potentiometric and impedimetric detection modes including portable, non-invasive, wireless, and self-driven miniaturized devices for monitoring human and animal health. Diagnostics of selected free radical precursors, low molecular biomarkers, nucleic acids and protein-based biomarkers, bacteria and viruses of today's interest is demonstrated.

Evaluation of antioxidant activity of chlorogenic acids and coffee extracts by an electrochemical DNA-based biosensor.

Sensitivity of dsDNA structure towards OH radicals as the pro-oxidants has been utilized as the detection principle of an analytical procedure applied for the first time to the evaluation of antioxidant activity (AOA) of 6 chlorogenic acids (CGAs) and extracts of 10 coffees. A nanostructured electrochemical DNA-based biosensor was prepared using a commercial electrode assembly and treated in the DNA cleavage agent formed by the Fenton type reaction. An addition of CGAs and aqueous coffee extracts significantly diminishes the degree of DNA degradation determined using cyclic voltammetry (CV) with the redox indicator [Fe(CN)6]3-/4-. The AOA decreases in order caffeic acid, CFA, >caffeoylquinic acids, CQAs, >dicaffeoylquinic acids, diCQAs, exhibiting the relative portion of survived DNA of about 71%, 70% and 69%, respectively, and of about 72% for C. robusta, Cherry, India (green bean) to 49% for Nescafé Espresso. Mechanisms of antioxidative properties are discussed.

Interaction of DNA and mononucleotides with theophylline investigated using electrochemical biosensors and biosensing.

The understanding of DNA-drug interaction mechanism is among the important aspects of biological studies for drug design, discovery and pharmaceutical development processes. Published rather detailed FTIR and UV-visible spectroscopic studies on the interactions of theophylline, theobromine and caffeine with calf thymus DNA have shown effective binding of these methylxanthine derivatives to DNA and RNA involving H-bonds. However, to our knowledge, there is no such investigation using electrochemical approach. As a novelty of the study, in this paper the bioelectrochemical approach has been chosen for the investigation of an interaction of low molecular salmon sperm dsDNA, ssDNA and mononucleotides with theophylline (TP) in aqueous phosphate buffered medium using DNA-based electrochemical biosensors and biosensing in solution phase. Exploitation of the electrochemical approach via changes in square wave voltammetric responses of deoxyguanosine (dGuo) and deoxyadenosine (dAdo) provided a new indication on preferential association of TP with dGuo in the case of double helical dsDNA structure which was not reported previously. Moreover, an attachment of TP molecules outside DNA was found in the presence of high concentration of 3.3 × 10-4 M TP in solution which diminishes the electron transfer and leads to the difficulties in quantitative evaluation of the TP and dGuo voltammetric responses. The changes in UV-vis and FTIR spectra obtained in the same medium confirmed the association interaction of TP with both nucleobases. Utilizing the model and the published energies of hydrogen bonding stabilization, the formation of a DNA-TP complex was predicted through the intermolecular H-bonds between TP and the NH-CO moiety of guanine and the N-NH2 moiety of adenine.

Current trends in electrochemical sensing and biosensing of DNA methylation.

DNA methylation plays an important role in physiological and pathological processes. Several genetic diseases and most malignancies tend to be associated with aberrant DNA methylation. Among other analytical methods, electrochemical approaches have been successfully employed for characterisation of DNA methylation patterns that are essential for the diagnosis and treatment of particular diseases. This article discusses current trends in the electrochemical sensing and biosensing of DNA methylation. Particularly, it provides an overview of applied electrode materials, electrode modifications and biorecognition elements applications with an emphasis on strategies that form the core DNA methylation detection approaches. The three main strategies as (i) bisulfite treatment, (ii) cleavage by restriction endonucleases, and (iii) immuno/affinity reaction were described in greater detail. Additionally, the availability of the reviewed platforms for early cancer diagnosis and the approval of methylation inhibitors for anticancer therapy were discussed.

Electrochemical and optical study of metallothionein interactions with prion proteins.

The prion protein (PrPC) can be structurally shifted to its PrPSc isoform causing a wide range of neurodegenerative diseases, which are currently incurable. There is an evidence that metallothioneins (MTs), and especially MT-3, are associated with neurodegenerative diseases. PrPC and MTs play pivotal roles in maintaining metal homeostasis; therefore, it is conceivable that each of them has its own significance in prion diseases. In this paper, we study the nature of interactions between PrPC, MT, and copper ions, Cu(II), using the method of differential pulse voltammetry (DPV) coupled with adsorptive transfer stripping technique (AdTS). Electrochemical properties of PrP itself and its interactions with both the Cu(II) ions and MTs have been found. Based on the results obtained, we hypothesised the formation of the complex in molar ratio 2:1 (PrPC:MT). Surface plasmon resonance imaging (SPRi) was used as a control reference assay to further confirm results obtained by the electrochemical approach, such as the specific interactions between PrPC and MT-3.

Chemical Modification of Boron-Doped Diamond Electrodes for Applications to Biosensors and Biosensing.

Boron-doped diamond (BDD) is a prospective electrode material that possesses many exceptional properties including wide potential window, low noise, low and stable background current, chemical and mechanical stability, good biocompatibility, and last but not least exceptional resistance to passivation. These characteristics extend its usability in various areas of electrochemistry as evidenced by increasing number of published articles over the past two decades. The idea of chemically modifying BDD electrodes with molecular species attached to the surface for the purpose of creating a rational design has found promising applications in the past few years. BDD electrodes have appeared to be excellent substrate materials for various chemical modifications and subsequent application to biosensors and biosensing. Hence, this article presents modification strategies that have extended applications of BDD electrodes in electroanalytical chemistry. Different methods and steps of surface modification of this electrode material for biosensing and construction of biosensors are discussed.

An integrated phenol 'sensoremoval' microfluidic nanostructured platform.

Phenol is a widely used chemical that for several reasons may be released into the environment and, consequently, its detection and subsequent destruction into the ground and surface waters are of special importance. Herein, a simple lab-on-a-chip (LOC) device based on biocompatible and biodegradable CaCO3- poly(ethyleneimine) (PEI) nanostructured microparticles (MPs) to detect and remove phenolic wastes is proposed. The detection of phenol using a hybrid polydimethylsiloxane (PDMS)/glass chronoimpedimetric microchip and its removal in the same LOC system through the use of an extra CaCO3-PEI MPs microcolumn is achieved. For the first time, the chronoimpedance technique is applied in a LOC system for phenol sensing in a range of 0.01-10 µM achieving the limit of detection (LOD) of 4.64 nM. Moreover, this device shows a high repeatability with a relative standard deviation of 3% which is almost 4 times lower than that for the chronoamperometry technique. This LOC system represents an integrated platform for phenol sensing and removal (sensoremoval) that can be easily fabricated and is of a low cost, disposable and amenable to mass production.

Nanostructured CaCO3-poly(ethyleneimine) microparticles for phenol sensing in fluidic microsystem.

A new and simple strategy based on nanostructured CaCO3-poly(ethyleneimine) (PEI) microparticles (MPs) for phenol sensing using PDMS/glass fluidic microchip is developed. This fluidic microsystem including integrated screen-printed electrodes modified with CaCO3-PEI MPs and tyrosinase (Tyr) through cross-linking with glutaraldehyde, represents a low-cost platform for phenol detection. The designed fluidic microsystem improves the sensitivity of the biosensor allowing the detection of very low concentrations of phenol (up to 10 nM). This device shows high repeatability and low detection limit, is easy to be fabricated, inexpensive, disposable, and amenable to mass production.

Voltammetric detection of damage to DNA caused by nitro derivatives of fluorene using an electrochemical DNA biosensor.

An electrochemical DNA biosensor based on the screen printed carbon paste electrode (SPCPE) with an immobilized layer of calf thymus double-stranded DNA has been used for in vitro investigation of the interaction between genotoxic nitro derivatives of fluorene (namely 2-nitrofluorene and 2,7-dinitrofluorene) and DNA. Two types of DNA damage have been detected at the DNA/SPCPE biosensor: first, that caused by direct association of the nitrofluorenes, for which an intercalation association has been found using the known DNA intercalators [Cu(phen)(2)](2+) and [Co(phen)(3)](3+) as competing agents, and, second, that caused by short-lived radicals generated by electrochemical reduction of the nitro group (observable under specific conditions only).

Disposable DNA biosensor with the carbon nanotubes-polyethyleneimine interface at a screen-printed carbon electrode for tests of DNA layer damage by quinazolines.

A screen-printed carbon working electrode within a commercially available screen-printed three-electrode assembly was modified by using a composite of multiwalled carbon nanotubes (MWCNT) dispersed in polyethylenimine (PEI) followed by covering with the calf thymus dsDNA layer. Several electrochemical methods were used to characterize the biosensor and to evaluate damage to the surface-attached DNA: square wave voltammetry of the [Ru(bpy)(3)](2+) redox indicator and mediator of the guanine moiety oxidation, cyclic voltammetry and electrochemical impedance spectroscopy in the presence of the [Fe(CN)(6)](3-/4-) indicator in solution. Due to high electroconductivity and large surface area of MWCNT and positive charge of PEI, the MWCNT-PEI composite is an advantageous platform for the DNA immobilization by the polyelectrolyte complexation and its voltammetric and impedimetric detection. In this respect, the MWCNT-PEI interface exhibited better properties than the MWCNT-chitosan one reported from our laboratory previously. A deep DNA layer damage at incubation of the biosensor in quinazoline solution was found, which depends on the quinazoline concentration and incubation time.

Disposable electrochemical biosensor with multiwalled carbon nanotubes-chitosan composite layer for the detection of deep DNA damage.

A novel electrochemical DNA-based biosensor for the detection of deep DNA damage was designed employing the bionanocomposite layer of multiwalled carbon nanotubes (MWNT) in chitosan (CHIT) deposited on a screen printed carbon electrode (SPCE). The biocomponent represented by double-stranded (ds) herring sperm DNA was immobilized on this composite using layer-by-layer coverage to form a robust film. Individual and complex electrode modifiers are characterized by a differential pulse voltammetry (DPV) with the DNA redox marker [Co(phen)(3)](3+), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) with [Fe(CN)(6)](3-) as a redox probe in a phosphate buffer solution (PBS). A good correlation between the CV and EIS parameters has been found, thus confirming a strong effect of MWNT on the enhancement of the electroconductivity of the electrode surface and that of CHIT on the MWNT distribution at the electrode surface. Differences between the CV and EIS signals of the electrodes without and with DNA are used to detect deep damage to DNA, advantageously using simple working procedures in the same experiment.

Interaction of tin(II) and arsenic(III) with DNA at the nanostructure film modified electrodes.

Biosensors based on DNA and DNA-carbon nanotubes film immobilized at the surface of a screen-printed carbon electrode were used for simple in vitro tests of chemical toxicity. The damage to DNA caused by tin(II) and arsenic(III) compounds as components of specific reaction media was evaluated by means of an electrochemical DNA marker, [Co(phen)3](3+), as the portion of original dsDNA which survives an incubation of the biosensor in the cleavage medium. The results were confirmed by the electrically heated electrode and by the measurement of the DNA guanine moiety signal.

Nanostructured electrochemical DNA biosensors for detection of the effect of berberine on DNA from cancer cells.

Multi walled carbon nanotubes (MWNT) in dimethylformamide (DMF) or aqueous sodium dodecyl sulfate (SDS) solution, colloidal gold nanoparticles (GNP) in phosphate buffer solution (PBS), and a GNP-MWNT mixture in aqueous SDS solution have been investigated for chemical modification of a screen-printed carbon electrode used as the signal transducer of a dsDNA-based biosensor. Differential pulse voltammetry of the DNA redox marker Co[(phen)3]3+ and the guanine moiety anodic oxidation and cyclic voltammetry with K3[Fe(CN)6] as indicator revealed substantial enhancement of the response of the biosensor, particularly when MWNT in SDS solution was used. The biosensor was used in testing of berberine, an isoquinoline plant alkaloid with significant antimicrobial and anticancer activity. Berberine had a very strong, concentration-dependent, effect on the structural stability of DNA from the human cancer cells (U937 cells) whereas non-cancer cells were changed only when berberine concentrations were relatively high 75 and 50 microg mL(-1).

Cytotoxicity and detection of damage to DNA by 3-(5-nitro-2-thienyl)-9-chloro-5-morpholin-4-yl[1,2,4]triazolo[4,3-c] quinazoline on human cancer cell line HeLa.

Quinazolines - 1,3-benzodiazines are biological active compounds, which are used in the phamaceutical industry, in agriculture and in the medicine. As documented in the literature, many derivatives demonstrated anticancer activity and they act as multitarget agents. 3-(5-Nitro-2-thienyl)-9-chloro-5-morpholin-4-yl[1,2,4]triazolo[4,3-c] quinazoline (NTCHMTQ) - a new synthetically prepared quinazoline derivative was the most effective derivative in our primary cytotoxic screening. In this study, we evaluated cytotoxic/antiproliferative activity of NTCHMTQ using human tumor cell line HeLa. Possible interaction of 3-(5-nitro-2-thienyl)-9-chloro-5-morpholin-4-yl[1,2,4]triazolo[4,3-c] quinazoline with calf thymus DNA was tested by the DNA - modified screen - printed electrode. Quinazoline derivative acted cytotoxically on tumor cell line HeLa. The IC(100) value was 10 microg/ml. The IC(50) values was found to be less than 4 microg/ml, a limit put forward by the National Cancer Institute (NCI) for classification of he compound as a potential anticancer drug. Quinazoline at micromolar concentrations induced morphological changes and necrosis of HeLa cells. Using the DNA based electrochemical biosensor, we have not found damage to DNA under in vitro conditions at an incubation of the biosensor in mixture with quinazoline.

Evaluation of the redox properties and anti/pro-oxidant effects of selected flavonoids by means of a DNA-based electrochemical biosensor.

Quercetin and rutin as well as catechin and epigallocatechin gallate were investigated, as widely distributed representatives of flavonols and flavanols, respectively, regarding their anti/pro-oxidant properties. The flavonoids are irreversibly oxidized at a dsDNA-modified screen-printed electrode within 0.368 to 0.449 V vs. SHE without binding to DNA. Using the DNA biosensor the detection scheme of a DNA prevention/degradation exploits the [Co(phen)(3)](3+) complex as an electrochemical DNA marker. Antioxidant activity of flavonoids was tested in a model cleavage mixture composed of 5 x 10(-7) mol L(-1) [Cu(phen)(2)](2+) as the catalyst, 1 x 10(-3) mol L(-1) ascorbic acid as the chemical reductant and atmospheric oxygen as the natural oxidant where reactive oxygen radicals are generated. The antioxidant activity increases with the concentration of flavonoids reaching a maximum where pro-oxidative behaviour becomes of importance. The pro-oxidant potency of flavonoids depends on the presence of atmospheric oxygen and follows the order quercetin>rutin>epigallocatechin gallate>catechin.

Detection of damage to DNA and antioxidative activity of yeast polysaccharides at the DNA-modified screen-printed electrode.

A simple procedure for the voltammetric detection of the DNA damage and antioxidants protecting DNA from its damage using a disposable electrochemical DNA biosensor is reported. The carbon-based screen-printed electrode (SPE) modified by a surface layer of the calf thymus double stranded (ds) DNA was used as a working electrode in combination with a silver/silver chloride reference electrode and a separate platinum auxiliary electrode. The [Co(phen)(3)](3+) ion served as the dsDNA redox marker and the [Cu(phen)(2)](2+) and [Fe(EDTA)](-) complex compounds were used as the DNA cleavage agents under the reduction by a chemical reductant (ascorbic acid). Four yeast polysaccharides with different chemical structure were investigated as the antioxidants within the concentration range of 0.05-4 mg ml(-1) in the cleavage mixture. A remarkable antioxidative activity of polysaccharides in order mannan (Candida krusei)>extracellular glucomannan (Candida utilis)>mannan (Candida albicans)>glucomannan (C. utilis) was found which is in agreement with that refered to trolox (a structural derivative of alpha-tocopherol) and determined by photochemiluminescent method.