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Pesticides are largely used at worldwide level to improve food production, fulfilling the needs of the global population which is increasing year by year. Although pesticides are beneficial for crop production, their extensive use has serious consequences for the pollution of the produced food as well as for soil and groundwaters. Indeed, it is reported that 50% of sprayed pesticides reach different destinations other than their target species, including soil, surface waters, and groundwaters. For this reason, we developed a flower-like origami paper-based device for pesticides detection in aerosol phase for precision agriculture. In detail, the paper-based electrochemical platform detects paraoxon, 2,4-dichlorophenoxyacetic acid, and glyphosate at ppb levels by measuring their inhibitory activity towards three different enzymes namely butyrylcholinesterase, alkaline phosphatase, and peroxidase enzyme, respectively. This integrated electrochemical device is composed of three office paper-based screen-printed electrodes and filter paper-based pads loaded with enzymes and enzymatic substrates. The pesticide detection is carried out by measuring through chronoamperometric technique the initial and residual enzymatic activity by using a smartphone-assisted potentiostat and evaluating the percentage of inhibition, proportional to the amount of aerosolized pesticides. This paper-based device was able to detect the three classes of pesticides in aerosol phase with limits of detection equal to 30 ppb, 10 ppb, and 2 ppb, respectively for 2,4-D, glyphosate, and paraoxon.

Current studies are investigating new E-cadherin gene (CDH1) mutations that may be responsible for diffuse gastric cancer susceptibility. Recently, a novel CDH1 germline variant presenting a G/A nucleotide change at cDNA position 2494 has been found in Japanese patients with familial diffuse gastric cancer. The consequent amino acid variation (Val/Met) may alter the binding activity to beta-catenin and the adhesive function of the E-cadherin protein. We have investigated its frequency in Italian cases of sporadic diffuse gastric cancer a well as in healthy controls. Peripheral blood samples were collected from consecutive patients with sporadic, diffuse gastric cancer and from healthy controls in the District of Urbino, Marche Region, Central Italy. After DNA extraction, standard techniques for molecular analyses were used to investigate the 2494 G/A germline nucleotide change in CDH1 cDNA. None of the 48 patients and 48 controls showed the G/A 2494 nucleotide change. Assuming a binomial distribution of the mutation among individuals and the absence of mutations in the 48 patients, the 95% upper bound for the underlying mutation frequency was 7.4%. The novel CDH1 nucleotide change is uncommon in Italian patients with sporadic diffuse gastric cancer. Given these results, further analyses in large population-based studies are not advisable

To assess the value of Computed Tomography (CT) in the diagnosis and in morphologic characterization of metastatic melanoma.The data of total body CT of 124 consecutive patients with melanoma having a Breslow index 1 mm or a positive sentinel lymph node have been retrospectively reviewed.The CT scan showed loco-regional and/or distant metastases in 36 patients (39%). Ten of these (28%) had metastases only to lymph nodes, whereas 26 patients (72%) had multiple metastases. Nodal, pulmonary, brain, subcutaneous, hepatic, adrenal, bone, gastrointestinal, breast and abdominal wall metastases were detected in 80.6%, 47.2%, 25%, 25%, 16.7%, 13.9%, 11.1%, 5.6%, 5.6% and 2.8% of the patients respectively. All the patients with metastases also had a positive sentinel lymph nodes and/or symptoms of metastatic disease.CT fails to reveal any characteristic feature of metastatic melanoma, but it is of value in the diagnosis of loco-regional and distant metastases in III stage disease.

The growth of (bio)sensors in analytical chemistry is mainly attributable to the development of affordable, effective, portable, and user-friendly analytical tools. In the field of sensors, paper-based devices are gaining a relevant position for their outstanding features including foldability, ease of use, and instrument-free microfluidics. Herein, a multifarious use of filter paper to detect copper ions in bodily fluids is reported by exploiting this eco-friendly material to (i) synthesize AuNPs without the use of reductants and/or external stimuli, (ii) print the electrodes, (iii) load the reagents for the assay, (iv) filter the gross impurities, and (v) preconcentrate the target analyte. Copper ions were detected down to 3 ppb with a linearity up to 400 ppb in standard solutions. The applicability in biological matrices, namely, sweat and serum, was demonstrated by recovery studies and by analyzing these biofluids with the paper-based platform and the reference method (atomic absorption spectroscopy), demonstrating satisfactory accuracy of the novel eco-designed analytical tool.

Paper-based devices are always more gaining a relevant position in the field of sensors. The continuous demand for affordable, simple, sustainable, and portable devices, is making paper as the ideal basis towards the realization of analytical tools for the easy self-testing. In this work, we demonstrate, for the first time, the development of a disposable paper-based printed electroanalytical strip for reliable, rapid, and high-throughput detection of glutathione in blood. The detection is based on the thiol-disulfide exchange reaction, which produces a detectable compound easily oxidizable at a Prussian Blue/carbon black nanocomposite involving a favorable low-interference overpotential. This nanocomposite is mixed within a carbon-based conductive ink and successively screen-printed onto a wax-patterned filter paper. The employment of paper provides a reagent-free device, as a consequence of the reagents pre-loading within the testing area. After the experimental conditions have been optimized, glutathione has been detected up to 10 mM, with a detection limit of 60 μM, and a sensitivity of (0.102 ± 0.005) μA/mM. This sensor showed satisfactory repeatability (relative standard deviation equal to 10%, for detection of glutathione 1 mM), especially by considering the hand-made manufacturing process. The “real-world” applicability of this strip has been evaluated by quantifying blood glutathione at physiological levels and by recovery studies achieving satisfactory values.

Despite substantial advances in sensing technologies, the development, preparation, and use of self-testing devices is still confined to specialist laboratories and users. Decentralized analytical devices will enormously impact daily lives, enabling people to analyze diverse clinical, environmental, and food samples, evaluate them and make predictions to improve quality of life, particularly in remote, resource-scarce areas. In recent years, paper-based analytical tools have attracted a great deal of attention; the well-known properties of paper, such as abundance, affordability, lightness, and biodegradability, combined with features of printed electrochemical sensors, have enabled the development of sustainable devices that drive (bio)sensors beyond the state of the art. Their blindness toward colored/turbid matrices (i.e., blood, soil), their portability, and the capacity of paper to autonomously filter/purge/react with target species make such devices powerful in establishing point-of-need tools for use by non-specialists. This protocol describes the preparation of a voltammetric phosphate sensor and an amperometric nerve agent biosensor; both platforms produce quantitative measurements with currents in the range of microamperes. These printed strips comprise three electrodes (graphite for working and counter electrodes and silver/silver chloride (Ag/AgCl) for the reference electrode) and nanomodifiers (carbon black and Prussian blue) to improve their performance and specificity. Depending on analytical need, different types of paper (filter, office) and configurations (1D, 2D, 3D) can be adopted. The protocol, based on the use of cost-effective manufacturing techniques such as drop casting (to chemically modify the substrate surface) and wax/screen printing (for creating the channels and electrodes), can be completed in

Nowadays, environmentally friendly synthesis pathways for preserving the environment and minimizing waste are strongly required. Herein, we propose filter paper as a convenient scaffold for chemical reactions. To demonstrate this novel approach, Prussian Blue Nanoparticles (PBNPs) were synthesized on filter paper by utilizing few μL of its precursors without external inputs, i.e. pH, voltage, reducing agents, and without producing waste as well. The functional paper, named “Paper Blue”, is successfully applied in the sensing field, exploiting the reduction of hydrogen peroxide at low applied potential. The eco-designed “Paper Blue” was combined with wax- and screen-printing to manufacture a reagentless electrochemical point-of-care device for diabetes self-monitoring, by using glucose oxidase as the biological recognition element. Blood glucose was linearly detected for a wide concentration range up to 25 mM (450 mg/dL), demonstrating its suitability for management of diabetes and glucose-related diseases. The Paper Blue-based biosensor demonstrated a correlation coefficient of 0.987 with commercial glucose strips (Bayer Contour XT). The achieved results demonstrated the effectiveness of this approach, which is also extendible to other (bio)systems to be applied in catalysis, remediation, and diagnostics.

Altres ajuts: the ICN2 is funded by the CERCA Programme/Generalitat de Catalunya. The development of paper-based electroanalytical strips as powerful diagnostic tools has gained a lot of attention within the sensor community. In particular, the detection of nucleic acids in complex matrices represents a trending topic, especially when focused toward the development of emerging technologies, such as liquid biopsy. DNA-based biosensors have been largely applied in this direction, and currently, there are two main approaches based on target/probe hybridization reported in the literature, namely Signal ON and Signal OFF. In this technical note, the two approaches are evaluated in combination with paper-based electrodes, using a single strand DNA relative to H1047R (A3140G) missense mutation in exon 20 in breast cancer as the model target. A detailed comparison among the analytical performances, detection protocol, and cost associated with the two systems is provided, highlighting the advantages and drawbacks depending on the application. The present work is aimed to a wide audience, particularly for those in the field of point-of-care, and it is intended to provide the know-how to manage with the design and development stages, and to optimize the platform for the sensing of nucleic acids using a paper-based detection method.

Biosensor development exploiting various transduction principles is characterized by a strong competition to reach high detectability, portability and robustness. Nevertheless, a literature-based comparison is not possible, as different conditions are employed in each paper. Herein, we aim at evaluating which measurement, photons or electrons, yields better biosensor performance. Upon outlining an update in recent achievements to boost analytical performance, amperometry and chemiluminescence (CL)-based biosensors are directly compared employing the same biospecific reagents and analytical formats. Horseradish peroxidase (HRP) and hydrogen peroxide concentrations were directly measured, while glucose and mouse IgG were detected employing an enzyme paper-based biosensor and an immunosensor, respectively. Detectability was down to picomoles of hydrogen peroxide (4 pmol for CL and 210 pmol for amperometry) and zeptomoles of HRP (45 zmol for CL and 20 zmol for amperometry); IgG was detected down to 12 fM (CL) and 120 fM (amperometry), while glucose down to 17 μM (CL) and 40 μM (amperometry). Results showed that amperometric and CL biosensors offered similar detectability and analytical performance, with some peculiarities that suggest complementary application fields. As they generally provided slightly higher detectability and wider dynamic ranges, CL-based biosensors appear more suitable for point-of-care testing of clinical biomarkers, where detectability is crucial. Nevertheless, as high detectability in CL biosensors usually requires longer acquisition times, their rapidity will allocate electrochemical biosensors in real-time monitoring and wearable biosensors. The analytical challenge demonstrated that these biosensors have competitive and similar performance, and between photons and electrons the competition is still open.

The detection of double stranded DNA (dsDNA) is often associated with the use of laboratory-bound approaches and/or with the prior generation of single stranded DNA (ssDNA), making these methods not suitable for in situ monitoring, i.e., point-of-care diagnostics. Screen-printed technology, coupled to the use of triplex forming oligonucleotides (TFO) as the recognizing probes, offers a great possibility toward the development of portable analytical tools. Moreover, the continuous demand for sustainable processes and waste lowering have highlighted the role of paper-based substrates for manufacturing easy-to-use, low-cost, and sustainable electrochemical devices. In this work, filter paper and copy paper have been utilized to produce E-DNA strips. Gold nanoparticles (AuNPs) have been exploited to immobilize the methylene blue (MB)-tagged TFO and to enhance the charge transfer kinetics at the electrode surface. Both paper-based substrates have been electrochemically characterized, and in addition, the effect of the amount of waxed layers has been evaluated. The paper-based E-DNA strips have been challenged toward the detection of three model targets, obtaining 3 and 7 nM as the detection limit, respectively, for single and double stranded sequences. The repeatability of the manufacturing (homemade) process has been evaluated with a relative standard deviation of approximately 10%. The effectiveness of the filter paper-based platform has been also evaluated in undiluted serum obtaining a similar value of the detection limit (compared to the measurements carried out in buffer solution). In addition, a synthetic PCR amplified dsDNA sequence, related to HIV, has been detected in serum samples.

The development of portable and user-friendly sensing platforms is a hot topic in the field of analytical chemistry. Among others, electroanalytical approaches exhibit a high amenability for reaching this purpose, i.e. the commercial strips for diabetes care are an obvious success. However, providing fully-integrated and reagent-free methods is always a leitmotiv. In this work, we evaluated the use of a disposable pipette tip, opportunely configured to demonstrate the first example of an electrochemical biosystem in a pipette tip, namely bio-lab-on-a-tip. The combination of a pipette tip, wire electrodes, enzyme, and cotton wool filter, allows the fabrication of a novel electroanalytical platform that does not need expertise-required tasks. To demonstrate the feasibility of this novel method, glucose is detected in beverages by means of chronoamperometry. The experimental setup, entirely built inside the pipette tip, is able to 1) block impurities/interferences from matrix, 2) load/release reagents for the bio-assay, 3) reduce the operating task to zero, and 4) perform electrochemical detection. With optimized experimental parameters, the bio-lab-on-a-tip is able to detect glucose linearly up to 10 mM with a detection limit of 170 μM. The effectiveness of the platform was confirmed by testing commercial beverages, e.g. Coca-Cola and Coca-Cola Zero, with high accuracy. In addition, the shelf-life of the novel device was evaluated, highlighting the role of cotton wool filter for providing a suitable environment for glucose oxidase stability. The novel concept can be easily generalized for further applications in the field of non-invasive clinical diagnostics and in-situ environmental monitoring.

Herein we describe a novel reagentless paper-based electrochemical phosphate sensor, manufactured with a simple and inexpensive approach. By following three easy steps, consisting of wax patterning, paper chemical modification, and electrode screen-printing, the filter paper provides an effective electroanalytical platform to sense phosphate ions in standard solutions and real samples (river water). The electrochemical properties of the paper-based platform were evaluated, firstly, by using ferricyanide as a redox mediator, proving no analyte-entrapment due to the cellulose lattice. Then, the reference colorimetric method for phosphate ions, which is based on the formation of phosphomolybdic complex, was successfully adapted to a reagentless electrochemically paper-based platform. This novel and highly sustainable configuration readily allows for the determination of phosphate ions with high reproducibility and long storage stability, achieving a detection limit of 4 μM over a wide linear range up to 300 μM. This in-house approach would be able to generically develop an affordable in situ and user-friendly sensing device without the addition of any reagent, to be applied for a broad range of analytes.

A facile ⿿one-step⿿ route to produce a homogenous and highly stable cobalt phthalocyanine (CoPc)-based dispersion by using carbon black (CB) as supporting material is reported. Herein, CB is proposed as effective material to load CoPc in order to obtain a CB/CoPc hybrid nanocomposite dispersion suitable for modifying screen-printed electrodes (SPEs) by an easy and automatable drop casting approach. CoPc resulted anchored to CB by a non-covalent physisorption, confirmed by IR and UV-visible spectroscopies, allowing to preserve the electrochemical performances of CoPc. The resulting CB/CoPc-modified SPE was tested as sensing tool to detect thiocholine, an enzymatic product of butyrylcholinesterase (BChE). The use of CB/CoPc leads to a highly sensitive thiocholine detection by applying a low potential (+0.05 V vs. internal reference) without fouling problem, a typical drawback that affects the thiol electrochemical detection. The favorable characteristics of the sensor were exploited for an easy BChE biosensor fabrication that renders this biosensor well suitable for mass-production. This electrochemical monoenzymatic biosensor was then challenged towards paraoxon, chosen as model organophosphorous pesticide, obtaining a low detection limit (18 nM). The suitability of the biosensor was tested in a waste water sample obtaining satisfactory recovery values, thus demonstrating its capability in such complex matrix.

A screen-printed electrode (SPE) modified with a carbon black (CB)-Au nanoparticles (AuNPs) composite is assembled and tested. Electrochemical and morphological investigations highlight the physico-chemical properties of the resulting AuNP-CB-SPE amperometric device with respect to SPEs modified with a single component of the nanocomposite. The effective performance of such a modified electrode in activating electrocatalytic processes, consisting both in oxidation and reduction reactions, is demonstrated. In particular, electrochemical tests on analytes such as glucose, hydrogen peroxide, hydroquinone, and ascorbic acid, evidence that the composite possesses electrocatalytic performance well superior with respect to the relevant mono-component modified SPE. As a consequence, a meaningful lowering of the peak potentials and improvement of the sensor sensitivities is observed when using AuNP-CB-SPEs with respect to both CB-SPEs and AuNP-SPEs. In the case of H2O2 reduction, the occurrence of the electrochemical process at less negative potentials is coupled to an improvement of sensor sensitivity of about one order of magnitude. Concurrently, lower limit of detections, ranging from 20 to 99% less, have been obtained for the major part of the analytes studied, i.e. glucose, hydrogen peroxide and hydroquinone. Preliminary results reported here indicate that AuNP-CB-SPE can be proposed as an efficient amperometric sensor to be used in many analytical applications.

Bioassays based on enzyme inhibition are analytical tools widely employed for inhibitor analysis. Beside the conventional analytical techniques such as chromatography and mass spectrometry, these bioassays are cost-effective, easy to use, and suitable for in situ measurement but they are often characterised by a quite narrow linear range. Herein, we report a novel graphical method based on integrated Michaelis-Menten equation, valid for all types of reversible inhibition, which provides an extended linear range. The suitability of this innovative approach was demonstrated in the case of fluoride quantification using a colorimetric bioassay based on acetylcholinesterase inhibition. The “half time reaction”, estimated by the progress curve of cholinesterase inhibition, was plotted versus the fluoride inhibitor concentration, observing an extended linear range up to 5 mM, instead of 0.6 mM using initial rate measurements. The applicability of this new concept was further demonstrated in the case of catalase enzyme inhibited by cyanide. Furthermore, it was demonstrated that fixed substrate conversion at level of 10–50% allows determination of inhibitor concentration in a wide linear range with high precision and in short time of analysis. This novel theoretical and practical approach allows for the extension of the linear range without any further experiments, with several advantages including low reagent consumption, reduced waste generation and time of measurement.

Graphene has been emphasized by researchers as the most promising nanomaterial for many application fields, including energy, catalysis, electronics, remediation, and sensing. In the field of analytic sciences, the continuous demand of sensitive, portable, user-friendly, low-cost, and low-volume analyses has highlighted graphene and graphene-related nanocomposites as key components toward the implementation of graphene-based (bio)sensors. This chapter provides the opportunity to summarize and critically evaluate the recent developments of graphene-based electrochemical devices that have been applied in biomedical field, for the detection of a wide variety of analytes of interest ranging from small compounds to nucleic acids, antibodies, proteins, and bacteria. Particular focus will interest graphene-based lab-on-a-chip (LOC) devices that have been (or will be) capable to answer the important query introduced by the World Health Organization (WHO) toward the development of affordable, sensitive, specific, user-friendly, rapid and robust, equipment-free and deliverable to end users (ASSURED) devices and to replace traditional approaches in the biomedical self-diagnostic field.

The recent goal of sustainability in analytical chemistry has boosted the development of eco-designed analytical tools to deliver fast and cost-effective analysis with low economic and environmental impact. Due to the recent focus in sustainability, we report the use of low-cost filter paper as a sustainable material to print silver electrodes and to load reagents for a reagent-free electrochemical detection of chloride in biological samples, namely serum and sweat. The electrochemical detection of chloride ions was carried out by exploiting the reaction of the analyte (i.e. chloride) with the silver working electrode. During the oxidation wave in cyclic voltammetry the silver ions are produced, thus they react with chloride ions to form AgCl, while in the reduction wave, the following reaction occurs: AgCl + e- -->Ag + Cl-. These reactions at the electrode surface resulted in anodic/cathodic peaks directly proportional to the chloride ions in solution. Chloride ions were detected with the addition of only 10 μL of the sample on the paper-based electrochemical cell, obtaining linearity up to 200 mM with a detection limit equal to 1 mM and relative standard deviation lower than 10%. The accuracy of the sensor was evaluated in serum and sweat samples, with percentage recoveries between 93 ± 10 and 108 ± 8%. Moreover, the results achieved with the paper-based device were positively compared with those obtained by using the gold standard method (Ion Selective Electrode) adopted in routine clinical analyses.

Herein, we report the first example of a paper-based screen-printed biosensor for the detection of ethanol in beer samples. Common office paper was adopted to fabricate the analytical device. The properties of this paper-based screen-printed electrode (SPE) were investigated by cyclic voltammetry, electrochemical impedance spectroscopy, and scanning electron microscopy, and they were compared with the well-established polyester-based SPEs as well. Paper demonstrated similar properties when compared with polyester, highlighting suitability towards its utilization in sensor development, with the advantages of low cost and simple disposal by incineration. A nanocomposite formed by Carbon Black (CB) and Prussian Blue nanoparticles (PBNPs), namely CB/PBNPs, was utilized as an electrocatalyst to detect the hydrogen peroxide generated by the enzymatic reaction between alcohol oxidase (AOx) and ethanol. After optimizing the analytical parameters, such as pH, enzyme, concentration, and working potential, the developed biosensor allowed a facile quantification of ethanol up to 10 mM (0.058 %vol), with a sensitivity of 9.13 μA/mM cm2 (1574 μA/%vol cm2) and a detection limit equal to 0.52 mM (0.003%vol). These satisfactory performances rendered the realized paper-based biosensor reliable over the analysis of ethanol contained in four different types of beers, including Pilsner, Weiss, Lager, and alcohol-free. The proposed manufacturing approach offers an affordable and sustainable tool for food quality control and for the realization of different electrochemical sensors and biosensors as well.

A novel sensor based on carbon black-gold nanoparticle nanocomposite modified screen-printed electrode (CB-AuNPs/SPE) for the detection of As(III) has been developed. The sensor was prepared modifying the SPE with CB and AuNPs by a drop casting automatable deposition. The As(III) was detected by CB-AuNPs/SPE using anodic stripping voltammetry, with a high sensitivity (673 6 mAmM 1cm 2) and reaching a LOD of 0.4 ppb. Finally, CB-AuNPs/SPE has been applied to As(III) trace analysis in drinking water, obtaining satisfactory recovery values (99 9%).

K.S. Novoselov in his Nobel lecture (December 8, 2010), described graphene as "more than just a flat crystal" and summarized the best possible impression of graphene with (i) it is the first example of 2D atomic crystals, (ii) it demonstrated unique electronic properties, thanks to charge carriers which mimic massless relativistic particles, and (iii) it has promise for a number of applications. The fascinating and unusual properties of this 2D material were indeed recently investigated and exploited in several disciplines including physics, medicine, and chemistry, indicating the extremely versatile and polyedric aspect of this nanomaterial. The utilization of nanomaterials, printed technology, and microfluidics in electroanalysis has resulted in a period that can be called the "Electroanalysis Renaissance" (Escarpa, 2012) in which graphene is without any doubt a forefront nanomaterial. The rise in affordable fabrication processes, along with the great dispersing attitude in a plenty of matrices, have made graphene powerful in large-scale production of electrochemical platforms. Herein, we overview the employment of graphene to customize and/or fabricate printable based (bio)sensors over the past 5 years, including several modification approaches such as drop casting, screen- and inkjet-printing, different strategies of graphene-based sensing, and applications as well. The objective of this review is to provide a critical perspective related to advantages and disadvantages of using graphene in biosensing tools, based on screen-printed sensors.

Different tyrosinase carbon paste modified electrodes to determine bisphenol A (BPA) concentration in aqueous solutions have been constructed. Variables examined were in the carbon paste composition and in particular: (i) the immobilized enzyme amount; (ii) the carbon type (powder, single or multi-walled nanotubes); (iii) the nature of the pasting oil (mineral oil, hexadecane and dodecane). For each biosensor type the amperometric response was evaluated with reference to the linear range and sensitivity. Constant reference has been made to the amperometric signals obtained, under the same experimental conditions, towards the catechol, a specific phenolic substrate for tyrosinase. The most efficient biosensors were those constructed by using the following composition for the carbon paste: 10% of tyrosinase, 45% of single wall carbon nanotubes (SWCN) and 45% of mineral oil. This biosensor formulation displayed the following electrochemical characteristics: a sensitivity equal to 138 μA/mM, LOD of 0.02 μM (based on three times the S/N ratio), linear range of 0.1–12 μM and response time of 6 min. This experimental work represents a first attempt at construction of a new carbon nanotube-tyrosinase based biosensor able to determine the concentration of BPA, one of the most ubiquitous and hazardous endocrine disruptors which can pollute the drinking and surface water, as well as many products of the food chain.