Autolytic attempt mimicking Brugada type 1 electrocardiogram pattern due to flecainide toxicity. A case report.

Background: Brugada phenocopies are a group of heterogeneous disorders that mimic Brugada syndrome (BrS) electrocardiogram (ECG) changes elicited by reversible clinical conditions. We report a novel case on flecainide toxicity causing an ECG signature of Brugada type 1 pattern in the paediatric age. Case summary: A 13-year-old Caucasian boy with untreated attention-deficit/hyperactivity disorder referred to the Pediatric Emergency Department (PED) after unknown antiarrhythmic drug overdose. He deliberately ingested 10 tablets from a labelled white box of a 100-mg single dose. The ECG showed a coved-type ST-segment elevation in right precordial leads and prolongation of PR segment with a QTc limit interval. Values of troponins gradually increased and echocardiogram was normal. The altered ECG pattern was explained by the stabilizing membrane effect of flecainide involving the inhibition of rapid Na+ channels. After offending drug removal, regression of ECG changes was observed and no cardiac events were documented during follow-up. Discussion: Flecainide-induced Brugada type 1 ECG pattern may occur in patients with no evidence of genetic susceptibility receiving a toxic dosage of this drug. With increasing dose, its action on conduction pathways manifests as prolongation of PR interval and QT and QRS complex duration and may cause BrS mimicry. A detailed clinical history considering symptoms and ECG findings may support early-raised suspicion for flecainide ingestion. The therapeutic approach implies primary detoxification, prevention of potential triggers, and management of eventual cardiotoxicity events. Finally, risk stratification for BrS should be always measured according to the clinical scenario and surveillance considered in a timely manner.

Reduced Graphene Oxide Electrolyte-Gated Transistor Immunosensor with Highly Selective Multiparametric Detection of Anti-Drug Antibodies.

The advent of immunotherapies with biological drugs has revolutionized the treatment of cancers and auto-immune diseases. However, in some patients, the production of anti-drug antibodies (ADAs) hampers the drug efficacy. The concentration of ADAs is typically in the range of 1-10 pm; hence their immunodetection is challenging. ADAs toward Infliximab (IFX), a drug used to treat rheumatoid arthritis and other auto-immune diseases, are focussed. An ambipolar electrolyte-gated transistor (EGT) immunosensor is reported based on a reduced graphene oxide (rGO) channel and IFX bound to the gate electrode as the specific probe. The rGO-EGTs are easy to fabricate and exhibit low voltage operations (≤ 0.3 V), a robust response within 15 min, and ultra-high sensitivity (10 am limit of detection). A multiparametric analysis of the whole rGO-EGT transfer curves based on the type-I generalized extreme value distribution is proposed. It is demonstrated that it allows to selectively quantify ADAs also in the co-presence of its antagonist tumor necrosis factor alpha (TNF-α), the natural circulating target of IFX.

RecruitNet: A global database of plant recruitment networks.

Plant recruitment interactions (i.e., what recruits under what) shape the composition, diversity, and structure of plant communities. Despite the huge body of knowledge on the mechanisms underlying recruitment interactions among species, we still know little about the structure of the recruitment networks emerging in ecological communities. Modeling and analyzing the community-level structure of plant recruitment interactions as a complex network can provide relevant information on ecological and evolutionary processes acting both at the species and ecosystem levels. We report a data set containing 143 plant recruitment networks in 23 countries across five continents, including temperate and tropical ecosystems. Each network identifies the species under which another species recruits. All networks report the number of recruits (i.e., individuals) per species. The data set includes >850,000 recruiting individuals involved in 118,411 paired interactions among 3318 vascular plant species across the globe. The cover of canopy species and open ground is also provided. Three sampling protocols were used: (1) The Recruitment Network (RN) protocol (106 networks) focuses on interactions among established plants ("canopy species") and plants in their early stages of recruitment ("recruit species"). A series of plots was delimited within a locality, and all the individuals recruiting and their canopy species were identified; (2) The paired Canopy-Open (pCO) protocol (26 networks) consists in locating a potential canopy plant and identifying recruiting individuals under the canopy and in a nearby open space of the same area; (3) The Georeferenced plot (GP) protocol (11 networks) consists in using information from georeferenced individual plants in large plots to infer canopy-recruit interactions. Some networks incorporate data for both herbs and woody species, whereas others focus exclusively on woody species. The location of each study site, geographical coordinates, country, locality, responsible author, sampling dates, sampling method, and life habits of both canopy and recruit species are provided. This database will allow researchers to test ecological, biogeographical, and evolutionary hypotheses related to plant recruitment interactions. There are no copyright restrictions on the data set; please cite this data paper when using these data in publications.

Spectroelectrochemical Enzyme Sensor System for Acetaldehyde Detection in Wine.

A new spectroelectrochemical two-enzyme sensor system has been developed for the detection of acetaldehyde in wine. A combination of spectroscopy and electrochemistry improves the analytical features of the electrochemical sensor because the optical information collected with this system is only associated with acetaldehyde and avoids the interferents also present in wines as polyphenols. Spectroelectrochemical detection is achieved by the analysis of the optical properties of the K3[Fe(CN)6]/K4[Fe(CN)6] redox couple involved in the enzymatic process: aldehyde dehydrogenase catalyzes the aldehyde oxidation using ß-nicotinamide adenine dinucleotide hydrate (NAD+) as a cofactor and, simultaneously, diaphorase reoxidizes the NADH formed in the first enzymatic process due to the presence of K3[Fe(CN)6]. An analysis of the characteristic UV-vis bands of K3[Fe(CN)6] at 310 and 420 nm allows the detection of acetaldehyde, since absorption bands are only related to the oxidation of this substrate, and avoids the contribution of other interferents.

Selective Ion Sensing in Artificial Sweat Using Low-Cost Reduced Graphene Oxide Liquid-Gated Plastic Transistors.

Health monitoring is experiencing a radical shift from clinic-based to point-of-care and wearable technologies, and a variety of nanomaterials and transducers have been employed for this purpose. 2D materials (2DMs) hold enormous potential for novel electronics, yet they struggle to meet the requirements of wearable technologies. Here, aiming to foster the development of 2DM-based wearable technologies, reduced graphene oxide (rGO)-based liquid-gated transistors (LGTs) for cation sensing in artificial sweat endowed with distinguished performance and great potential for scalable manufacturing is reported. Laser micromachining is employed to produce flexible transistor test patterns employing rGO as the electronic transducer. Analyte selectivity is achieved by functionalizing the transistor channel with ion-selective membranes (ISMs) via a simple casting method. Real-time monitoring of K+ and Na+ in artificial sweat is carried out employing a gate voltage pulsed stimulus to take advantage of the fast responsivity of rGO. The sensors show excellent selectivity toward the target analyte, low working voltages (<0.5 V), fast (5-15 s), linear response at a wide range of concentrations (10 µm to 100 mm), and sensitivities of 1 µA/decade. The reported strategy is an important step forward toward the development of wearable sensors based on 2DMs for future health monitoring technologies.

Immobilization of Antibodies by Genetic Fusion to a Fungal Self-Assembling Adhesive Protein.

Although antibody immobilization on solid surfaces is extensively used in several applications, including immunoassays, biosensors, and affinity chromatography, some issues are still challenging. Self-assembling protein layers can be used to coat easily different surfaces by direct deposition. A specific biofunctional layer can be formed using genetic engineering techniques to express fused proteins acting as self-immobilizing antibodies. In this study, fusion proteins combining the self-assembling adhesive properties of a fungal hydrophobin and the functionality of the single chain fragment variables (ScFvs) of two antibodies were produced. The chosen ScFvs are able to recognize marine toxins associated with algal blooms, saxitoxin, and domoic acid, which can bioaccumulate in shellfish and herbivorous fish causing food poisoning. ScFvs fused to hydrophobin Vmh2 from Pleurotus ostreatus were produced in Escherichia coli and recovered from the inclusion bodies. The two fusion proteins retained the functionality of both moieties, being able to adhere on magnetic beads and to recognize and bind the two neurotoxins, even with different performances. Our immobilization procedure is innovative and very easy to implement because it allows the direct functionalization of magnetic beads with ScFvs, without any surface modification. Two different detection principles, electrochemical and optical, were adopted, thus achieving a versatile platform suitable for different antigen detection methods. The sensitivity of the saxitoxin optical biosensor [limit of detection (LOD) 1.7 pg/ml] is comparable to the most sensitive saxitoxin immunosensors developed until now.

Detection of dithiocarbamate, chloronicotinyl and organophosphate pesticides by electrochemical activation of SERS features of screen-printed electrodes.

Enhancement of Raman intensity due to the electrochemical surface-enhanced Raman scattering (EC-SERS) effect is an interesting alternative to overcome the lack of sensitivity traditionally associated with Raman spectroscopy. Furthermore, activation of metallic screen-printed electrodes (SPEs) by electrochemical route leads to the reproducible generation of nanostructures with excellent SERS properties. EC-SERS procedure proposed in this work for the detection of several pesticides (thiram, imidacloprid and chlorpyrifos) with different nature, uses gold SPEs as SERS substrates, but also includes a preconcentration step as the initial and essential stage. Taking into account the small volume of solution employed, only 60 µL, the preconcentration cannot be performed for more than 15 min in order to ensure the proper contact of the solution with WE, RE and CE. Furthermore, selected temperature, 34 °C, is not very high to allow the exhaustive control of the drop volume. Optimization of preconcentration parameters (time and temperature) displays a crucial step, particularly in the detection of low concentrations of pesticides, because it will provide higher Raman intensity in EC-SERS experiments. After the initial step, gold SPEs were electrochemically activated by cyclic voltammetry, allowing the detection of very low concentration (µg·L-1) of pesticides due to the generation of fresh nanostructures with SERS effect.

Understanding the ECL interaction of luminol and Ru(bpy)32+ luminophores by spectro-electrochemiluminescence.

A detailed analysis of the ECL interaction between luminol and tris(2,2'-bipyridyl)dichlororuthenium(ii) (Ru(bpy)32+) is required before using them in ECL systems for multianalyte detection purposes. Spectro-electrochemiluminescence demonstrates that not only must the emission properties be considered, but also their additional optical characteristics are involved in the explanation of the interaction mechanism between these luminophores.

Screen-Printed Electrodes Modified with Metal Phthalocyanines: Characterization and Electrocatalysis in Chlorinated Media.

Metal phthalocyanines are well-known sensing phases with applications in different scientific fields due to their interesting properties. Detailed characterization by Raman spectroscopy was performed in order to study the shifting of the vibrational bands related to the coordination sphere of each metal phthalocyanine. In this work, a study involving the use of screen-printed electrodes (SPEs) with various metal phthalocyanines to electrochemically detect and quantify chlorine (Cl2) gas is presented. The Cl2 gas was generated in-situ via oxidation of the chloride present in form of aqueous salt solutions. The developed method offers not only the possibility to quantify chlorine, but also to discriminate among several chlorinated species due to the changes observed in the voltammetric profiles associated with the interaction between the specie assayed and the phthalocyanine metallic center. Optimization of detecting parameters was also performed to apply this procedure for the quantification of chlorine generated from commercial chlorine tablets. The development of this proof of concept shows interesting possibilities and easy-to-use applications with novel on metal phthalocyanines based SPE sensors.

Screen-Printed Electrodes Modified with Metal Nanoparticles for Small Molecule Sensing.

Recent progress in the field of electroanalysis with metal nanoparticle (NP)-based screen-printed electrodes (SPEs) is discussed, focusing on the methods employed to perform the electrode surface functionalization, and the final application achieved with different types of metallic NPs. The ink mixing approach, electrochemical deposition, and drop casting are the usual methodologies used for SPEs' modification purposes to obtain nanoparticulated sensing phases with suitable tailor-made functionalities. Among these, applications on inorganic and organic molecule sensing with several NPs of transition metals, bimetallic alloys, and metal oxides should be highlighted.

Spectroelectrochemical elucidation of B vitamins present in multivitamin complexes by EC-SERS.

Raman spectroelectrochemistry based on electrochemical surface-enhanced Raman scattering (EC-SERS) effect is an interesting alternative to overcome the lack of sensitivity of normal Raman spectroscopy. Electrochemical activation of metallic screen-printed electrodes (SPEs) leads to the reproducible generation of nanostructures with excellent SERS properties. In that way, gold SPEs circumvent the traditional reproducibility limitation and produce the enhancement of the Raman intensity to favor the detection of low concentrations. Furthermore, fingerprint features of Raman spectroscopy make possible the dynamic spectroelectrochemical analysis of B vitamins. The accuracy assignments of Raman bands associated with B1, B2, B3, B6 and B12 vitamins present in multivitamin complexes provides valuable information, allowing us not only the detection of B vitamin present in mixtures, but also to understand the interaction between vitamins and metallic SERS surfaces.

Sex and the cost of reproduction through the life course of an extremely long-lived herb.

Despite being central concepts for life history theory, little is known about how reproductive effort and costs vary with individual age once plants have started to reproduce. We conducted a 5-year field study and estimated age-dependent reproductive effort for both sexes in the extraordinarily long-lived dioecious plant Borderea pyrenaica. We also evaluated costs of reproduction on vital rates for male and female plants, both by examining effects of differences in individual reproductive effort under natural conditions, and by conducting a flower removal experiment, aimed at decreasing reproductive effort. Reproductive effort was fairly constant and independent of age for males, which may reflect a strategy of adjusting overall reproductive output by spreading reproduction over the life course. Females had a higher total effort, which first increased and then decreased with age. The latter may be a response to an increasing reproductive value-an inverse of a terminal investment-or a sign of reproductive senescence due to an age-related physiological decline. Seed production was lower in plants with higher previous reproductive effort and this effect increased with age. We found no evidence for costs of reproduction on other vital rates for either sex. Experimental flower removal only resulted in progressively more negative effects on flower production in older male plants, whereas female vital rates were unaffected. Overall, this study demonstrates that not only sex, but also age influences resource allocation trade-offs and, thus, plant life history evolution.

Resolution of mixed dyes by in situ near infrared (NIR) spectroelectrochemistry.

NIR spectroelectrochemistry has scarcely been used for deconvolving aqueous mixtures due to the water restriction in this spectral range. However, this work offers an interesting approach for the study of mixtures of molecules with similar electrochemical and spectroscopic behaviour by overcoming the limitations of this hybrid technique. As a proof of concept, the resolution of mixtures of two dyes with similar chemical structures demonstrates the usefulness of NIR spectroelectrochemistry.

Towards single-molecule in situ electrochemical SERS detection with disposable substrates.

Dynamic time-resolved Raman spectroelectrochemistry demonstrates the strong influence of nanostructuring and surface charge of in situ activated disposable substrates for SERS detection. Under specific conditions, a large enhancement factor and estimated calculations agree with the feasible detection of only a few molecules, approaching the limit of single-entity detection.

In Situ Spectroelectrochemical Monitoring of Dye Bleaching after Electrogeneration of Chlorine-Based Species: Application to Chloride Detection.

Spectroelectrochemical techniques are becoming increasingly versatile tools to solve a diverse range of analytical problems. Herein, the use of in situ real-time luminescence spectroelectrochemistry to quantify chloride ions is demonstrated. Utilizing the bleaching effect of chlorine-based electrogenerated products after chloride oxidation, it is shown that the fluorescence of the rhodamine 6G dye decreases proportionally to the initial chloride concentration in solution. A strong decrease of fluorescence is observed in acidic media compared to a lower decrease in alkaline media, which suggests that Cl2, favorably generated at low pH, could be the main species responsible for the fluorescence loss. This fact is corroborated with chronoamperometric measurements where the complete loss of fluorescence for the bulk solution is achieved. A fast mass transfer is needed to explain this behavior, in agreement with the generation of gaseous species such as Cl2. Chloride detection was performed in artificial sweat samples in less than 30 s with great accuracy. This electrochemical/optical combined approach allows us to quantify species that are difficult to measure by electrochemistry due to the inadequate resolution of their redox processes or being without significant optical properties.

Quenching of graphene quantum dots fluorescence by alkaline phosphatase activity in the presence of hydroquinone diphosphate.

In this work, a turn-off photoluminescent sensing proof-of-concept based on blue luminescent graphene quantum dots (GQDs) as the fluorescent probe was developed. For that purpose, GQDs optical response was related with the catalytic enzymatic activity of alkaline phosphatase (ALP), in the presence of hydroquinone diphosphate (HQDP). The hydrolysis of HQDP by ALP generated hydroquinone (HQ). The oxidation of HQ, enzymatically produced, to p-benzoquinone (BQ) resulted in the quenching of GQDs fluorescence (FL). Therefore, the developed luminescent sensing mechanism allowed the FL quenching with ALP activity to be related and thus quantified the concentration of ALP down to 0.5 nM of enzyme. This innovative design principle appears as a promising tool for the development of enzymatic sensors based on ALP labeling with fluorescent detection or even for direct ALP luminescent quantification in an easy, fast and sensitive manner.

Evaluation of electrochemical, UV/VIS and Raman spectroelectrochemical detection of Naratriptan with screen-printed electrodes.

Naratriptan, active pharmaceutical ingredient with antimigraine activity was electrochemically detected in untreated screen-printed carbon electrodes (SPCEs). Cyclic voltammetry and differential pulse voltammetry were used to carry out quantitative analysis of this molecule (in a Britton-Robinson buffer solution at pH 3.0) through its irreversible oxidation (diffusion controlled) at a potential of +0.75V (vs. Ag pseudoreference electrode). Naratriptan oxidation product is an indole based dimer with a yellowish colour (maximum absorption at 320nm) so UV-VIS spectroelectrochemistry technique was used for the very first time as an in situ characterization and quantification technique for this molecule. A reflection configuration approach allowed its measurement over the untreated carbon based electrode. Finally, time resolved Raman Spectroelectrochemistry is used as a powerful technique to carry out qualitative and quantitative analysis of Naratriptan. Electrochemically treated silver screen-printed electrodes are shown as easy to use and cost-effective SERS substrates for the analysis of Naratriptan.

Novel thin layer flow-cell screen-printed graphene electrode for enzymatic sensors.

A new Screen-printed electrodes (SPE) integrated in one channel flow-cell was developed. The one channel flow-cell is attached and directly changeable with electrode. In the new flow-cell the injection is done through an "in-line luer injection port" which can be less aggressive than wall-jet flow cell for a biological recognition element immobilized on the surface of the electrode. The sample volume can be easily controlled by the operator through a syringe. In this novel thin layer flow-cell screen-printed electrodes, the working electrode was modified with graphene materials, and an enhancement of electroactive area to 388% over a standard electrode was found. This new configuration was applied to study the entrapped cellobiose dehydrogenase from the ascomycete Corynascus thermophilus (CtCDH) in a photocrosslinkable PVA-based polymer. The calibration curve of lactose using optimized parameters shows a wide linear measurement ranges between 0.25 and 5mM. A good operational stability of the CtCDH-PVA-modified graphene electrode is obtained, which keeps the same initial activity during 8h and exhibits a good storage stability with a decrease of only 9% in analytical response after 3 months storage at 4◦C.

Miniaturized analytical instrumentation for electrochemiluminescence assays: a spectrometer and a photodiode-based device.

Herein, a new miniaturized analytical instrumentation for electrochemiluminescence (ECL) assays is presented. A photodiode integrated in an ECL cell combined with a potentiostat/galvanostat, all integrated in a one-piece instrument (µSTAT ECL), was developed. In addition, a complementary micro-spectrometer integrated in a similar ECL cell for luminescence spectra recording is also proposed. Both cells are intended to be used with screen-printed electrodes and all the devices are portable and small sized. Their performance was corroborated with two innovative proofs-of-concept that centered on the luminol transduction chemistry: a first time reported ECL assay based on the enzymatic reaction between an indoxyl substrate and the enzyme alkaline phosphatase, and the electrochemiluminescence resonance energy transfer (ECL-RET) process triggered by the electro-oxidized luminol to the acceptor fluorescein. The photodiode system revealed to be more sensitive than the spectrometer device in collecting the light; however, with the latter, it is possible to discriminate different luminescent species according to their maximum wavelength emission, which is extremely useful for carrying out simple and simultaneous ECL multiplex analyzes. The spectrometer device works as an excellent accessory to couple with the µSTAT ECL instrument, complementing the experiments. Graphical abstract Schematic representation of the ECL-RET: from luminol-H2O2 system to fluorescein, the micro-spectrometer for the light collection and the 3D representation of the ECL-RET reaction.