Recent Achievements in Electrochemical and Optical Nucleic Acids Based Detection of Metal Ions.

Recently nucleic acids gained considerable attention as selective receptors of metal ions. This is because of the possibility of adjusting their sequences in new aptamers selection, as well as the convenience of elaborating new detection mechanisms. Such a flexibility allows for easy utilization of newly emerging nanomaterials for the development of detection devices. This, in turn, can significantly increase, e.g., analytical signal intensity, both optical and electrochemical, and the same can allow for obtaining exceptionally low detection limits and fast biosensor responses. All these properties, together with low power consumption, make nucleic acids biosensors perfect candidates as detection elements of fully automatic portable microfluidic devices. This review provides current progress in nucleic acids application in monitoring environmentally and clinically important metal ions in the electrochemical or optical manner. In addition, several examples of such biosensor applications in portable microfluidic devices are shown.

Portable Nitric Oxide (NO) Generator Based on Electrochemical Reduction of Nitrite for Potential Applications in Inhaled NO Therapy and Cardiopulmonary Bypass Surgery.

A new portable gas phase nitric oxide (NO) generator is described for potential applications in inhaled NO (INO) therapy and during cardiopulmonary bypass (CPB) surgery. In this system, NO is produced at the surface of a large-area mesh working electrode by electrochemical reduction of nitrite ions in the presence of a soluble copper(II)-ligand electron transfer mediator complex. The NO generated is then transported into gas phase by either direct purging with nitrogen/air or via circulating the electrolyte/nitrite solution through a gas extraction silicone fiber-based membrane-dialyzer assembly. Gas phase NO concentrations can be tuned in the range of 5-1000 ppm (parts per million by volume for gaseous species), in proportion to a constant cathodic current applied between the working and counter electrodes. This new NO generation process has the advantages of rapid production times (5 min to steady-state), high Faraday NO production efficiency (ca. 93%), excellent stability, and very low cost when using air as the carrier gas for NO (in the membrane dialyzer configuration), enabling the development of potentially portable INO devices. In this initial work, the new system is examined for the effectiveness of gaseous NO to reduce the systemic inflammatory response (SIR) during CPB, where 500 ppm of NO added to the sweep gas of the oxygenator or to the cardiotomy suction air in a CPB system is shown to prevent activation of white blood cells (granulocytes and monocytes) during extracorporeal circulation with cardiotomy suction conducted with five pigs.

Polyion selective polymeric membrane-based pulstrode as a detector in flow-injection analysis.

A method for the detection of polyions using fully reversible polyion selective polymeric membrane type pulstrodes as detectors in a flow-injection analysis (FIA) system is examined. The detection electrode consists of a plasticized polymeric membrane doped with 10 wt % of tridodecylmethylammonium-dinonylnaphthalene sulfonate (TDMA/DNNS) ion-exchanger salt. The pulse sequence used involves a short (1 s) galvanostatic pulse, an open-circuit pulse (0.5 s) during which the EMF of the cell is measured, and a longer (15 s) potentiostatic pulse to return the membrane to its original chemical composition. It is shown that total pulse sequence times can be optimized to yield reproducible real-time detection of injected samples of protamine and heparin at up to 20 samples/h. Further, it is shown that the same membrane detector can be employed for FIA detection of both polycations at levels ≥10 µg/mL and polyanions at levels of ≥40 µg/mL by changing the direction of the galvanostatic pulse. The methodology described may also be applicable in the detection of polyionic species at low levels in other flowing configurations, such as in liquid chromatography and capillary electrophoresis.

Toward the boosted loading of cisplatin drug into liposome nanocarriers.

Current challenges in oncology are largely associated with the need to improve the effectiveness of cancer treatment and to reduce drug's side effects. An effective strategy to cope with these challenges is behind designing and developing drug delivery systems based on smart nanomaterials and approved anticancer drugs. The present study offers a novel and straightforward approach to efficiently load the cisplatin drug into the newly constructed liposome-based nanosystems as well a reliable technique for monitoring this process based on capillary electrophoresis hyphenated with inductively coupled plasma tandem mass spectrometry. The proposed drug-loading methodology comprises liposome formation via a simple ethanol-injection method and propels increased drug encapsulation using tailor-made freeze-thawing or lyophilization-hydration procedures. To optimize liposome generation and drug encapsulation, the effects of dilution medium and liposome composition (types of phospholipids and their percentage ratio) have been investigated in detail. It was shown that modest alterations of the composition of three-component phospholipid liposomes and parameters of the freeze-thawing procedure have a strong impact on the formation of cisplatin-liposome systems. The obtained cisplatin-liposome formulation features a remarkable degree of drug encapsulation, over 100 mg L-1, and holds promise for further preclinical development as a potent drug-delivery platform.

Effective monitoring of Platinum-DNA adducts formation under simulated physiological conditions by CE-ICP-MS/MS.

The leading Pt(II)-based anticancer drugs have been used for decades; however, chemotherapy with their application is burdened with severe side effects. The administration of compounds capable of DNA platination in the form of prodrugs has the potential to overcome the drawbacks associated with their use. Progress toward their clinical application depends on establishing proper methodologies that would allow assessing their ability to bind to DNA in the biological environment. Herein, we propose implementing the approach based on the hyphenation of capillary electrophoresis with inductively coupled plasma tandem mass spectrometry (CE-ICP-MS/MS) for studying Pt-DNA adduct formation. The presented methodology opens the possibility to employ the multielement monitoring for studying the differences in the behavior of Pt(II) and Pt(IV) complexes and, interestingly, revealed the formation of various adducts with DNA and cytosol components for the latter one.

Drawbacks in the efficient monitoring of gold nanoparticle-based cisplatin delivery systems formation by HPLC-ICP-MS.

Since chemotherapy suffers many limitations related to side effects of anticancer drugs (e.g. cisplatin - CDDP), nanoparticles are probed as carriers in targeted drug delivery. Gold nanoparticles (AuNPs) are broadly investigated due to their biocompatibility, nontoxicity, and tunable surface. Despite many AuNPs-cisplatin systems (AuNP-CS) reports found in the literature, only a few include studies of their synthesis and formation efficiency using analytical tools providing simultaneously qualitative and quantitative analytical information. Therefore, this research continues our previous study of AuNP-CS formation investigated by capillary electrophoresis with inductively coupled plasma mass spectrometry (ICP-MS). Namely, it presents the analogical approach but employs the coupling of another separation technique: isocratic reversed-phase high-performance liquid chromatography. The study concerns the difficulties of analytical method optimization path and contains a discussion of the observed problematic issues related to the analysis and preparation of AuNP-CS. Moreover, the presented work confronts the performance and applicability of both tools for the scrutiny of AuNP-CS, especially considering the comparison of their resolution power.

How to effectively prepare a sample for bottom-up proteomic analysis of nanoparticle protein corona? A critical review.

Since the interest in the biomedical applications of inorganic nanoparticles (NPs) has rapidly grown over the last decades, there is a need for a thorough characterization of bio-nano interactions. NPs introduced to the body (mostly intravenously) encounter plasma proteins, that instantly create a so-called "protein corona" on the NPs surface, giving the nanomaterial a new biological identity. Type of the proteins that interact with NPs may affect the in vivo fate of NPs. For that reason, it is particularly important to establish analytical methods capable of corona protein identification. Bottom-up proteomics is most often used for that purpose. A crucial part of the experiment is sample preparation, as it is already proven that different protocols may lead to distinct results. This review is aimed at providing a characterization of two main stages of sample preparation: separation of NPs with protein corona from the unbound proteins and the digestion of corona proteins. Separation techniques such as centrifugation, magnetic separation, and chromatography and three digestion methods (in-gel, in-solution, and on-particle) are described with special emphasis paid on their advantages and disadvantages as well as their influence on the result of identification. This paper also indicates the need for standardization of protein corona identification protocols, as some of the proteins may be preferentially detected while applying a particular digestion procedure.

Comparison of electrochemical nitric oxide detection methods with chemiluminescence for measuring nitrite concentration in food samples.

Nitrite is a naturally occurring species present in various food samples and also present in our bodies as a product of nitric oxide (NO) oxidation. Considering the ubiquity of nitrite, its determination is of great importance in both biological and food samples. Herein, a very facile indirect method of nitrite determination in meat samples via selective reduction to nitric oxide (NO) is presented. The resulting gaseous product is quantified via portable and cost-effective electrochemical sensors. Both a novel laboratory prepared Pt-Nafion based NO sensor and a commercially available amperometric NO sensor are compared. Excellent correlations between the nitrite amount found in tested samples using both of the electrochemical sensors and a reference chemiluminescence method are demonstrated (r = 0.997 and r = 0.999 for Pt-Nafion based and commercially available NO-B4 electrochemical sensors, respectively, n = 12). Moreover, the slope of the linear regression curves are very close to unity for the comparison of the three systems tested. The amperometric sensors compared within this work exhibit good precision and accuracy and are shown to be an attractive alternative to the costly chemiluminescence detection method for accurately determining nitrite levels in food samples.

Performance of Amperometric Platinized-Nafion Based Gas Phase Sensor for Determining Nitric Oxide (NO) Levels in Exhaled Human Nasal Breath.

Nitric oxide (NO) levels in exhaled breath are a non-invasive marker that can be used to diagnose various respiratory diseases and monitor a patient's response to given therapies. A portable and inexpensive device that can enable selective NO concentration measurements in exhaled breath samples is needed. Herein, the performance of an amperometric Pt-Nafion-based gas phase sensor for detection of NO in exhaled human nasal breath is examined. Enhanced selectivity over carbon monoxide and ammonia is achieved via an in-line zinc oxide-based filter. Exhaled nasal NO levels measured in 21 human samples with the sensor are shown to correlate well with those obtained using a chemiluminescence reference method (R2 = 0.9836).

Electrochemical biosensor modified with dsDNA monolayer for restriction enzyme activity determination.

A simple and cost effective method for the determination of restriction endonuclease activity is presented. dsDNA immobilized at a gold electrode surface is used as the enzymatic substrate, and an external cationic redox probe is employed in voltammetric measurements for analytical signal generation. The assessment of enzyme activity is based on a decrease of a current signal derived from reduction of methylene blue which is present in the sample solution. For this reason, the covalent attachment of the label molecule is not required which significantly reduces costs of the analysis and simplifies the entire determination procedure. The influence of buffer components on utilized dsDNA/MCH monolayer stability and integrity is also verified. Electrochemical impedance spectroscopy measurements reveal that due to pinhole formation during enzyme activity measurement the presence of any surfactants should be avoided. Additionally, it is shown that the sensitivity of the electrochemical biosensor can be tuned by changing the restriction site location along the DNA length. Under optimal conditions the proposed biosensor exhibits a linear response toward PvuII activity within a range from 0.25 to 1.50 U/µL.