3D-printed holder for drawing highly reproducible pencil-on-paper electrochemical devices.

Pencil drawing is one of the simplest and most cost-effective ways of fabricating miniaturized electrodes on a paper substrate. However, it is limited by the lack of reproducibility regarding the electrode drawing process. A 3D-printed pencil holder (3DPH) is proposed here for simple, reproducible, and low-cost hand-drawn fabrication of paper-based electrochemical devices. 3DPH was designed to keep pressure and angulation of the graphite mine constant on the paper substrate using a micromechanical pencil regardless of the user/operator. This approach significantly improved the reproducibility and cost of making reliable pencil-drawn electrodes. The results showed high reproducibility and accuracy of the 3DPH-assisted electrodes prepared by 4 different operators in terms of sheet resistance and electrochemical behavior. Cyclic voltammetric (CV) curves in the presence of [Fe(CN)6]3-/4- redox probe showed only 3.9% variation for the anodic peak currents of different electrodes prepared by different operators when compared with electrodes prepared without the 3D-printed support. SEM analyses revealed a more uniform graphite deposition/design of the electrodes prepared with 3DPH, which corroborates the results obtained by CV. As a proof of concept, 3DPH-assisted pencil-drawn graphite electrodes were employed for dopamine detection in synthetic saliva, showing a proportional increase in anodic peak current at 0.12 V vs. carbon pRE with increasing dopamine (DA) concentration, with a detection limit of 0.39µmol L-1. Moreover recovery was in the range 93-104% of DA (4-7% RSD) in synthetic saliva for three different concentrations, demonstrating the reliability of the approach. Finally, we believe this approach can make pencil-drawn technology more robust, accessible, reliable, and inexpensive for real on-site applications, especially in hard-to-reach locations or research centers with little investment.

Zero-Biased Photoelectrochemical Detection of Cardiac Biomarker Myoglobin Based on CdSeS/ZnS Quantum Dots and Barium Titanate Perovskite.

Cardiovascular diseases are considered one of the leading causes of premature mortality of patients worldwide. Therefore, rapid diagnosis of these diseases is crucial to ensure the patient's survival. During a heart attack or severe muscle damage, myoglobin is rapidly released in the body to constitute itself as a precise biomarker of acute myocardial infarction. Thus, we described the photoelectrochemical immunosensor development to detect myoglobin. It was based on fluorine-doped tin oxide modified with CdSeS/ZnSe quantum dots and barium titanate (BTO), designated as CdSeS/ZnSQDS/BTO. It was characterized by scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDX), transmission electron microscopy (TEM), X-ray diffraction (XRD), electrochemical impedance spectroscopy (EIS), and amperometry. The anodic photocurrent at the potential of 0 V (vs. Ag/AgCl) and pH 7.4 was found linearly related to the myoglobin (Mb) concentration from 0.01 to 1000 ng mL-1. Furthermore, the immunosensor showed an average recovery rate of 95.7-110.7% for the determination of myoglobin.

Charge Storage in Graphene Oxide: Impact of the Cation on Ion Permeability and Interfacial Capacitance.

The charge storage and membrane applications of graphene oxide (GO) materials are dictated by its intrinsic material properties. Structure-function relationships correlating periodic parameters, such as the hydrated ion radius and ion-GO interactions, are currently lacking yet are needed to provide insight on the charge storage and ion transport mechanism. We report the use of scanning ion conductance microscopy to measure the ion permeability of GO films and evaluate its relationship with the measured capacitance. We demonstrate that species (namely K+) with strong electrostatic interactions with the oxygen functionalities of GO provide the benefit of higher capacitance but suffer from inhibited ion mobility due to constriction of the GO interlayer spacing.

Insight into the Electro-Oxidation Mechanism of Glucose and Other Carbohydrates by CuO-Based Electrodes.

In this work, a new hypothesis for the electrocatalytic behavior of CuO electrodes is presented. Different from the established mechanism, here we discuss why CuIII species do not participate in the oxidation mechanism of carbohydrates. We show that hydroxyl ion adsorption and the semiconductive properties of the material play a more significant role in this process. The relationship between the flat band potential and the potential that begin oxidation suggests that the concentration of vacancies in the charge region acts upon the reactivity of the adsorbed hydroxyl ions through a partial charge transfer reaction. In the presence of carbohydrate molecules, the electron transfer is facilitated and involves the transfer of electrons from the adsorbed hydroxyl ions to the CuO film. This mechanism is fundamentally relevant since it helps the understanding of several experimental misleads. The results can also lead to obtaining better catalysts, since improvements in the material should focus on enhancing the semiconductive properties rather than the CuII/CuIII redox transition. The results shed light on different aspects of carbohydrate molecules oxidation that could lead to novel applications and possibly a better description of other semiconductor mechanisms in electrocatalysis.

InP Nanowire Biosensor with Tailored Biofunctionalization: Ultrasensitive and Highly Selective Disease Biomarker Detection.

Electrically active field-effect transistors (FET) based biosensors are of paramount importance in life science applications, as they offer direct, fast, and highly sensitive label-free detection capabilities of several biomolecules of specific interest. In this work, we report a detailed investigation on surface functionalization and covalent immobilization of biomarkers using biocompatible ethanolamine and poly(ethylene glycol) derivate coatings, as compared to the conventional approaches using silica monoliths, in order to substantially increase both the sensitivity and molecular selectivity of nanowire-based FET biosensor platforms. Quantitative fluorescence, atomic and Kelvin probe force microscopy allowed detailed investigation of the homogeneity and density of immobilized biomarkers on different biofunctionalized surfaces. Significantly enhanced binding specificity, biomarker density, and target biomolecule capture efficiency were thus achieved for DNA as well as for proteins from pathogens. This optimized functionalization methodology was applied to InP nanowires that due to their low surface recombination rates were used as new active transducers for biosensors. The developed devices provide ultrahigh label-free detection sensitivities ∼1 fM for specific DNA sequences, measured via the net change in device electrical resistance. Similar levels of ultrasensitive detection of ∼6 fM were achieved for a Chagas Disease protein marker (IBMP8-1). The developed InP nanowire biosensor provides thus a qualified tool for detection of the chronic infection stage of this disease, leading to improved diagnosis and control of spread. These methodological developments are expected to substantially enhance the chemical robustness, diagnostic reliability, detection sensitivity, and biomarker selectivity for current and future biosensing devices.

Reusable, robust, and accurate laser-generated photonic nanosensor.

Developing noninvasive and accurate diagnostics that are easily manufactured, robust, and reusable will provide monitoring of high-risk individuals in any clinical or point-of-care environment. We have developed a clinically relevant optical glucose nanosensor that can be reused at least 400 times without a compromise in accuracy. The use of a single 6 ns laser (λ = 532 nm, 200 mJ) pulse rapidly produced off-axis Bragg diffraction gratings consisting of ordered silver nanoparticles embedded within a phenylboronic acid-functionalized hydrogel. This sensor exhibited reversible large wavelength shifts and diffracted the spectrum of narrow-band light over the wavelength range λpeak ≈ 510-1100 nm. The experimental sensitivity of the sensor permits diagnosis of glucosuria in the urine samples of diabetic patients with an improved performance compared to commercial high-throughput urinalysis devices. The sensor response was achieved within 5 min, reset to baseline in ∼10 s. It is anticipated that this sensing platform will have implications for the development of reusable, equipment-free colorimetric point-of-care diagnostic devices for diabetes screening.

Cation-Dependent Stabilization of Electrogenerated Naphthalene Diimide Dianions in Porous Polymer Thin Films and Their Application to Electrical Energy Storage.

Porous polymer networks (PPNs) are attractive materials for capacitive energy storage because they offer high surface areas for increased double-layer capacitance, open structures for rapid ion transport, and redox-active moieties that enable faradaic (pseudocapacitive) energy storage. Here we demonstrate a new attractive feature of PPNs--the ability of their reduced forms (radical anions and dianions) to interact with small radii cations through synergistic interactions arising from densely packed redox-active groups, only when prepared as thin films. When naphthalene diimides (NDIs) are incorporated into PPN films, the carbonyl groups of adjacent, electrochemically generated, NDI radical anions and dianions bind strongly to K(+), Li(+), and Mg(2+), shifting the formal potentials of NDI's second reduction by 120 and 460 mV for K(+) and Li(+)-based electrolytes, respectively. In the case of Mg(2+), NDI's two redox waves coalesce into a single two-electron process with shifts of 240 and 710 mV, for the first and second reductions, respectively, increasing the energy density by over 20 % without changing the polymer backbone. In contrast, the formal reduction potentials of NDI derivatives in solution are identical for each electrolyte, and this effect has not been reported for NDI previously. This study illustrates the profound influence of the solid-state structure of a polymer on its electrochemical response, which does not simply reflect the solution-phase redox behavior of its monomers.

Microemulsification: an approach for analytical determinations.

We address a novel method for analytical determinations that combines simplicity, rapidity, low consumption of chemicals, and portability with high analytical performance taking into account parameters such as precision, linearity, robustness, and accuracy. This approach relies on the effect of the analyte content over the Gibbs free energy of dispersions, affecting the thermodynamic stabilization of emulsions or Winsor systems to form microemulsions (MEs). Such phenomenon was expressed by the minimum volume fraction of amphiphile required to form microemulsion (Φ(ME)), which was the analytical signal of the method. Thus, the measurements can be taken by visually monitoring the transition of the dispersions from cloudy to transparent during the microemulsification, like a titration. It bypasses the employment of electric energy. The performed studies were: phase behavior, droplet dimension by dynamic light scattering, analytical curve, and robustness tests. The reliability of the method was evaluated by determining water in ethanol fuels and monoethylene glycol in complex samples of liquefied natural gas. The dispersions were composed of water-chlorobenzene (water analysis) and water-oleic acid (monoethylene glycol analysis) with ethanol as the hydrotrope phase. The mean hydrodynamic diameter values for the nanostructures in the droplet-based water-chlorobenzene MEs were in the range of 1 to 11 nm. The procedures of microemulsification were conducted by adding ethanol to water-oleic acid (W-O) mixtures with the aid of micropipette and shaking. The Φ(ME) measurements were performed in a thermostatic water bath at 23 °C by direct observation that is based on the visual analyses of the media. The experiments to determine water demonstrated that the analytical performance depends on the composition of ME. It shows flexibility in the developed method. The linear range was fairly broad with limits of linearity up to 70.00% water in ethanol. For monoethylene glycol in water, in turn, the linear range was observed throughout the volume fraction of analyte. The best limits of detection were 0.32% v/v water to ethanol and 0.30% v/v monoethylene glycol to water. Furthermore, the accuracy was highly satisfactory. The natural gas samples provided by the Petrobras exhibited color, particulate material, high ionic strength, and diverse compounds as metals, carboxylic acids, and anions. These samples had a conductivity of up to 2630 µS cm(-1); the conductivity of pure monoethylene glycol was only 0.30 µS cm(-1). Despite such downsides, the method allowed accurate measures bypassing steps such as extraction, preconcentration, and dilution of the sample. In addition, the levels of robustness were promising. This parameter was evaluated by investigating the effect of (i) deviations in volumetric preparation of the dispersions and (ii) changes in temperature over the analyte contents recorded by the method.

Ultradense Electrochemical Chips with Arrays of Nanostructured Microelectrodes to Enable Sensitive Diffusion-Limited Bioassays.

Nanostructured microelectrodes (NMEs) are an attractive alternative to yield sensitive bioassays in unprocessed samples. However, although valuable for different applications, nanoporous NMEs usually cannot boost the sensitivity of diffusion-limited analyses because of the enlarged Debye length within the nanopores, which reduces their accessibility. To circumvent this limitation, nanopore-free gold NMEs were electrodeposited from 45 µm SU-8 apertures, featuring nanoridged microspikes on a recessed surface of gold thin film while carrying interconnected crown-like and spiky structures along the edge of a SU-8 passivation layer. These structures were grown onto ultradense, vertical array chips that offer a promising strategy for translating reproducible, high-resolution, and cost-effective sensors into real-world applications. The NMEs yielded reproducible analyses, while machine learning allowed us to predict the analytical responses from NME electrodeposition data. By taking advantage of the high surface area and accessible structure of the NMEs, these structures provided a sensitivity for [Fe(CN)6]3-/4- that was 5.5× higher than that of bare WEs while also delivering a moderate antibiofouling property in undiluted human plasma. As a proof of concept, these electrodes were applied toward the fast (22 min) and simple determination of Staphylococcus aureus by monitoring the oxidation of [Fe(CN)6]4-, which acted as a cellular respiration rate redox reporter. The sensors also showed a wide dynamic range, spanning 5 orders of magnitude, and a calculated limit of detection of 0.2 CFU mL-1.

Single-Response Duplexing of Electrochemical Label-Free Biosensor from the Same Tag.

Multiplexing is a valuable strategy to boost throughput and improve clinical accuracy. Exploiting the vertical, meshed design of reproducible and low-cost ultra-dense electrochemical chips, the unprecedented single-response multiplexing of typical label-free biosensors is reported. Using a cheap, handheld one-channel workstation and a single redox probe, that is, ferro/ferricyanide, the recognition events taking place on two spatially resolved locations of the same working electrode can be tracked along a single voltammetry scan by collecting the electrochemical signatures of the probe in relation to different quasi-reference electrodes, Au (0 V) and Ag/AgCl ink (+0.2 V). This spatial isolation prevents crosstalk between the redox tags and interferences over functionalization and binding steps, representing an advantage over the existing non-spatially resolved single-response multiplex strategies. As proof of concept, peptide-tethered immunosensors are demonstrated to provide the duplex detection of COVID-19 antibodies, thereby doubling the throughput while achieving 100% accuracy in serum samples. The approach is envisioned to enable broad applications in high-throughput and multi-analyte platforms, as it can be tailored to other biosensing devices and formats.

Construction and electrochemical characterization of microelectrodes for improved sensitivity in paper-based analytical devices.

This work presents a simple, low cost method for creating microelectrodes for electrochemical paper-based analytical devices (ePADs). The microelectrodes were constructed by backfilling small holes made in polyester sheets using a CO2 laser etching system. To make electrical connections, the working electrodes were combined with silver screen-printed paper in a sandwich type two-electrode configuration. The devices were characterized using linear sweep voltammetry, and the results are in good agreement with theoretical predictions for electrode size and shape. As a proof-of-concept, cysteine was measured using cobalt phthalocyanine as a redox mediator. The rate constant (k(obs)) for the chemical reaction between cysteine and the redox mediator was obtained by chronoamperometry and found to be on the order of 10(5) s(-1) M(-1). Using a microelectrode array, it was possible to reach a limit of detection of 4.8 µM for cysteine. The results show that carbon paste microelectrodes can be easily integrated with paper-based analytical devices.

Sensing approaches on paper-based devices: a review.

Paper has been present in the world of analytical chemistry for centuries, but it seems that just a few years back it was rediscovered as a valuable substrate for sensors. We can easily list some of the countless advantages of this simple cellulosic substrate, including mechanical properties, three-dimensional fibrous structure, biocompatibility and biodegradability, easiness of production and modification, reasonable price, and availability all over the world. Those characteristics make paper a first-choice substrate for disposable sensors and integrated sensing platforms. Nowadays, numerous examples of paper-based sensors are being presented in the literature. This review describes some of the most prominent examples classifying them by type of detection: optical (colorimetric, fluorescence, surface-enhanced Raman spectroscopy, and transmittance methods) and electrochemical (voltammetric, potentiometric, and conductivity-based methods). We take a closer look at recent advances in immunoassays fabricated on paper, excluding simple lateral flow tests assembled on nitrocellulose. This review also summarizes the main advantages and disadvantages of the use of paper as a substrate for sensors, as well as its impact on their performance and application, presents a short history of paper in analytical chemistry, and discusses fabrication methods and available sources of paper.

Evaluation of a photoelectrochemical platform based on strontium titanate, sulfur doped carbon nitride and palladium nanoparticles for detection of SARS-CoV-2 spike glycoprotein S1.

This work aims to develop a photoelectrochemical (PEC) platform for detection of SARS-CoV-2 spike glyprotein S1. The PEC platform is based on the modification of a fluorine-doped tin oxide (FTO) coated glass slide with strontium titanate (SrTiO3 or ST), sulfur-doped carbon nitride (g-C3N4-S or CNS) and palladium nanoparticles entrapped in aluminum hydroxide matrix (PdAlO(OH) or PdNPs). The PEC platform was denoted as PdNPs/CNS/ST/FTO and it was characterized by SEM, TEM, FTIR, DRX, and EIS. The PEC response of the PdNPs/CNS/ST/FTO platform was optimized by evaluating the effects of the concentration of the donor molecule, the nature of the buffer, pH, antibody concentration, potential applied to the working electrode, and incubation time. The optimized PdNPs/CNS/ST/FTO PEC platform was modified with 5 µg mL-1 of antibody for determination of SARS-CoV-2 spike glycoprotein S1. A decrease in the photocurrent was observed with an increase in the concentration of SARS-CoV-2 from 1 fg mL-1 to 1000 pg mL-1 showing that the platform is a promising alternative for the detection of S1 protein from SARS-CoV-2. The designed PEC platform exhibited recovery percentages of 96.20% and 109.65% in artificial saliva samples.

Biocompatible Wearable Electrodes on Leaves toward the On-Site Monitoring of Water Loss from Plants.

Impedimetric wearable sensors are a promising strategy for determining the loss of water content (LWC) from leaves because they can afford on-site and nondestructive quantification of cellular water from a single measurement. Because the water content is a key marker of leaf health, monitoring of the LWC can lend key insights into daily practice in precision agriculture, toxicity studies, and the development of agricultural inputs. Ongoing challenges with this monitoring are the on-leaf adhesion, compatibility, scalability, and reproducibility of the electrodes, especially when subjected to long-term measurements. This paper introduces a set of sensing material, technological, and data processing solutions that overwhelm such obstacles. Mass-production-suitable electrodes consisting of stand-alone Ni films obtained by well-established microfabrication methods or ecofriendly pyrolyzed paper enabled reproducible determination of the LWC from soy leaves with optimized sensibilities of 27.0 (Ni) and 17.5 kΩ %-1 (paper). The freestanding design of the Ni electrodes was further key to delivering high on-leaf adhesion and long-term compatibility. Their impedances remained unchanged under the action of wind at velocities of up to 2.00 m s-1, whereas X-ray nanoprobe fluorescence assays allowed us to confirm the Ni sensor compatibility by the monitoring of the soy leaf health in an electrode-exposed area. Both electrodes operated through direct transfer of the conductive materials on hairy soy leaves using an ordinary adhesive tape. We used a hand-held and low-power potentiostat with wireless connection to a smartphone to determine the LWC over 24 h. Impressively, a machine-learning model was able to convert the sensing responses into a simple mathematical equation that gauged the impairments on the water content at two temperatures (30 and 20 °C) with reduced root-mean-square errors (0.1% up to 0.3%). These data suggest broad applicability of the platform by enabling direct determination of the LWC from leaves even at variable temperatures. Overall, our findings may help to pave the way for translating "sense-act" technologies into practice toward the on-site and remote investigation of plant drought stress. These platforms can provide key information for aiding efficient data-driven management and guiding decision-making steps.

A nanostructured piezoelectric immunosensor for detection of human cardiac troponin T.

A piezoelectric immunosensor based on gold nanoparticles (AuNPs) co-immobilized on a dithiol-modified surface is proposed for detection of human cardiac troponin T (TnT). Anti-human troponin T (anti-TnT) antibodies were covalently immobilized on the nanostructured electrode surface by thiol-aldehyde linkages. In a homogeneous bulk solution, TnT was captured by anti-TnT immobilized on the QCM electrode. Cyclic voltammetry studies were used to characterize the AuNPs layer on the electrode surface and the anti-TnT immobilization steps. The QCM-flow immunosensor exhibited good reliability, measuring concentrations of TnT from 0.003 to 0.5 ng mL(-1) in human serum with high linearity (r = 0.989; p < 0.01). The immunosensor exhibited a 7% coefficient of variation and 0.0015 ng mL(-1) limit of detection, indicating a high reproducibility and sensitivity. The proposed QCM nanostructured immunosensor is easy to use and has promising potential in the diagnosis of acute myocardial infarction due to its speed and high sensitivity.

Evaluation of PAMAM Dendrimers (G3, G4, and G5) in the Construction of a SPR-based Immunosensor for Cardiac Troponin T.

An immunosensor was developed using a SAM of an alkanethiol associated with PAMAM(G4) dendrimers based on surface plasmon resonance (SPR) to enhance the sensitivity for troponin T detection in blood samples. The feasibility of using three-dimensional platforms based on dendrimers for the development of immunosensors was demonstrated by evaluating three different generations of these dendrimers (G3, G4, and G5) to detect troponin T. The results showed the efficiency of these 3D platforms in anchoring biomolecules, amplifying the detection of troponin T. The sandwich assay showed good performance for troponin T detection, using secondary monoclonal antibodies, in the concentration range of 5 - 300 ng mL-1 (0.14 - 8.67 nmol L-1), R2 = 0.991, with the LOD of 3.6 ng mL-1. The sandwich assay's applicability was demonstrated by evaluating a secondary polyclonal antibody's performance in the concentration range of 3 - 30 ng mL-1, R2 = 0.998, with the LOD of 0.98 ng mL-1. The immunosensor was applied to determine troponin T in blood plasma samples from healthy patients, with an average recovery of 88 to 104%. The performance of the SPR-based immunosensor indicates reliable results and is expected to contribute to the rapid diagnosis of heart attack, with reduced costs.

Antigenotoxic potential of the fermentation broth produced by Paenibacillus polymyxa RNC-D in vitro.

Aim: Evaluate the chemopreventive potential of the extract from P. polymyxa RNC-D. Methods: Concentrations of P. polymyxa RNC-D extract were tested in HepG2/C3A cells to assess their genotoxic (comet assay), mutagenic (micronucleus test) and antigenotoxic potential (comet assay) in vitro. Results: 400 and 40 µg/ml concentrations induced DNA lesions, whereas the 4 µg/ml induced a desmutagenic effect. Complementary tests indicated that the extract minimized the formation of reactive oxygen species induced by methyl methanesulfonate and normalized the loss of membrane potential. The quantification of cytokines indicated that TNF-α was immunostimulated by the extract. However, when administered in conjunction with the methyl methanesulfonate, the extract blocked the TNF-α release. Conclusion: The fermentation broth from P. polymyxa RNC-D showed an antigenotoxic effect, and thus the potential to be used as chemopreventive compound.

Insights into the structure and function of the C-terminus of SGTs (small glutamine-rich TPR-containing proteins): A study of the Aedes aegypti homolog.

SGTs (small glutamine-rich TPR-containing proteins) are dimeric proteins that belong to the class of co-chaperones characterized by the presence of TPR domains (containing tetratricopeptide repeats). Human (SGTA) and yeast (Sgt2) SGTs are characterized by three distinct domains: an N-terminal dimerization domain, a central TPR-domain important for binding to other proteins (chaperones included) and a C-terminal domain involved in hydrophobic interactions. Both these SGTs are involved in the cellular PQC (protein quality control) system, as they interact with chaperones and have functions that aid stress recovery. However, there are differences between them, such as structural features and binding specificities, that could be better understood if other orthologous proteins were studied. Therefore, we produced and characterized a putative SGT protein, designated AaSGT, from the mosquito Aedes aegypti, which is a vector of several diseases, such as dengue and Zika. The protein was produced as a folded dimer which was stable up to 40 °C and was capable of binding to AaHsp90 and fully protecting a model protein, α-synuclein, from aggregation. The conformation of AaSGT was investigated by biophysical tools and small angle X-ray scattering, which showed that the protein had an elongated conformation and that its C-terminal domain was mainly disordered. The results with a C-terminal deletion mutant supported these observations. Altogether, these results are consistent with those from other functional SGT proteins and add to the understanding of the PQC system in Aedes aegypti, an important aim that may help to develop inhibitory strategies against this vector of neglected diseases.

On the structure and function of Sorghum bicolor CHIP (carboxyl terminus of Hsc70-interacting protein): A link between chaperone and proteasome systems.

The co-chaperone CHIP (carboxy terminus of Hsc70 interacting protein) is very important for many cell activities since it regulates the ubiquitination of substrates targeted for proteasomal degradation. However, information on the structure-function relationship of CHIP from plants and how it interacts and ubiquitinates other plant chaperones is still needed. For that, the CHIP ortholog from Sorghum bicolor (SbCHIP) was identified and studied in detail. SbCHIP was purified and produced folded and pure, being capable of keeping its structural conformation up to 42 °C, indicating that cellular function is maintained even in a hot environment. Also, SbCHIP was able to bind plant Hsp70 and Hsp90 with high affinity and interact with E2 enzymes, performing E3 ligase activity. The data allowed to reveal the pattern of plant Hsp70 and Hsp90 ubiquitination and described which plant E2 enzymes are likely involved in SbCHIP-mediated ubiquitination. Aditionally, we obtained information on the SbCHIP conformation, showing that it is a non-globular symmetric dimer and allowing to put forward a model for the interaction of SbCHIP with chaperones and E2 enzymes that suggests a mechanism of ubiquitination. Altogether, the results presented here are useful additions to the study of protein folding and degradation in plants.

Fabrication of microwell plates and microfluidic devices in polyester films using a cutting printer.

This study reports, for the first time, the possibility to manufacture analytical devices on polyester substrates using a cutting printer. The protocol involves the design of a layout in a graphical software, the cut into polyester films and the lamination against one or multiple polyester films coated with a thermosensitive layer. The feasibility of the proposed approach was demonstrated through the fabrication of 96-microwell plates, 3D microfluidic mixing and distance-based microfluidic devices. The printer has enabled cutting microchannels wider than 300 µm on polyester films and a thickness of 250 µm. Urea and glucose assays were performed on microwell plates aiming for their quantification in artificial urine and serum samples. The presented results revealed good agreement with the expected values. The complexation reaction between Fe2+ and o-phenanthroline was selected as model to investigate the feasibility of the 3D mixing device. Absorbance measurements were recorded for the reaction product performed in both on and off-chip modes. Considering the achieved data, the on-chip mixing exhibited similar behavior when compared to off-chip reaction, thus demonstrating to be efficient to perform mixtures due to the turbulence generated inside three-dimensional channels. Lastly, a distance-based device was designed to detect H2O2 based on the displacement of a dye plug promoted by the oxygen generation using a copper-modified paper sheet. The distance-based peroxymeter revealed a linear behavior in the concentration range between 1 and 5% (v/v) and a LOD equal to 0.5% (65.2 mM). Based on the results herein reported, the proposed method represents a simple and alternative protocol to produce microdevices, using affordable and inexpensive raw materials, within 10 min, and at a cost lower than US$ 0.10 per unit.