Graphene oxide fiber microelectrodes with controlled sheet alignment for sensitive neurotransmitter detection.

Here, we synthesized and characterized graphene oxide (GO) fiber microelectrodes with controllable nanosheet orientation to study the extent to which sheet alignment and orientation impacts electrochemical detection of neurochemicals. The alignment of the GO nanosheets was characterized by scanning electron microscopy, Raman spectroscopy, and cyclic voltammetry. The electrochemical performance of GO microelectrodes and its suitability for subsecond detection of neurotransmitters was further evaluated by fast-scan cyclic voltammetry (FSCV). We have shown that the GO sheet alignment has a considerable effect on the electron transfer kinetics, frequency independent behavior, and detection suitability for specific neurotransmitters. Therefore, this fine-tuning aspect of the electrode surface for specific electrochemical detection should be taken into consideration for any future utilization of GO-based biological sensors.

Distinguishing academic science writing from humans or ChatGPT with over 99% accuracy using off-the-shelf machine learning tools.

ChatGPT has enabled access to artificial intelligence (AI)-generated writing for the masses, initiating a culture shift in the way people work, learn, and write. The need to discriminate human writing from AI is now both critical and urgent. Addressing this need, we report a method for discriminating text generated by ChatGPT from (human) academic scientists, relying on prevalent and accessible supervised classification methods. The approach uses new features for discriminating (these) humans from AI; as examples, scientists write long paragraphs and have a penchant for equivocal language, frequently using words like "but," "however," and "although." With a set of 20 features, we built a model that assigns the author, as human or AI, at over 99% accuracy. This strategy could be further adapted and developed by others with basic skills in supervised classification, enabling access to many highly accurate and targeted models for detecting AI usage in academic writing and beyond.

Effects of carnosine on the embryonic development and TiO2 nanoparticles-induced oxidative stress on Zebrafish.

Oxidative stress is due to an unbalance between pro-oxidants, such as reactive oxygen (ROS) and nitrogen (RNS) species, and antioxidants/antioxidant system. Under physiological conditions these species are involved in different cellular processes such as cellular homeostasis and immune response, while an excessive production of ROS/RNS has been linked to the development of various diseases such as cancer, diabetes, and Alzheimer's disease. In this context, the naturally occurring dipeptide carnosine has shown the ability to scavenge ROS, counteract lipid peroxidation, and inhibit proteins oxidation. Titanium dioxide nanoparticles (TiO2-NPs) have been widely used to produce cosmetics, in wastewater treatment, in food industry, and in healthcare product. As consequence, these NPs are often released into aquatic environments. The Danio rerio (commonly called zebrafish) embryos exposure to TiO2-NPs did not affect the hatching rate, but induced oxidative stress. According to this scenario, in the present study, we first investigated the effects of carnosine exposure and of a sub-toxic administration of TiO2-NPs on the development and survival of zebrafish embryos/larvae measured through the acute embryo toxicity test (FET-Test). Zebrafish larvae represent a useful model to study oxidative stress-linked disorders and to test antioxidant molecules, while carnosine was selected based on its well-known multimodal mechanism of action that includes a strong antioxidant activity. Once the basal effects of carnosine were assessed, we then evaluated its effects on TiO2-NPs-induced oxidative stress in zebrafish larvae, measured in terms of total ROS production (measured with 2,7-dichlorodihydrofluorescein diacetate probe) and protein expression by immunohistochemistry of two cellular stress markers, 70 kDa-heat shock protein (Hsp70) and metallothioneins (MTs). We demonstrated that carnosine did not alter the phenotypes of both embryos and larvae of zebrafish at different hours post fertilization. Carnosine was instead able to significantly decrease the enhancement of ROS levels in zebrafish larvae exposed to TiO2-NPs and its antioxidant effect was paralleled by the rescue of the protein expression levels of Hsp70 and MTs. Our results suggest a therapeutic potential of carnosine as a new pharmacological tool in the context of pathologies characterized by oxidative stress such as neurodegenerative disorders.

Spatiotemporal Imaging of Zinc Ions in Zebrafish Live Brain Tissue Enabled by Fluorescent Bionanoprobes.

The zebrafish is a powerful model organism to study the mechanisms governing transition metal ions within whole brain tissue. Zinc is one of the most abundant metal ions in the brain, playing a critical pathophysiological role in neurodegenerative diseases. The homeostasis of free, ionic zinc (Zn2+) is a key intersection point in many of these diseases, including Alzheimer's disease and Parkinson's disease. A Zn2+ imbalance can eventuate several disturbances that may lead to the development of neurodegenerative changes. Therefore, compact, reliable approaches that allow the optical detection of Zn2+ across the whole brain would contribute to our current understanding of the mechanisms that underlie neurological disease pathology. We developed an engineered fluorescence protein-based nanoprobe that can spatially and temporally resolve Zn2+ in living zebrafish brain tissue. The self-assembled engineered fluorescence protein on gold nanoparticles was shown to be confined to defined locations within the brain tissue, enabling site specific studies, compared to fluorescent protein-based molecular tools, which diffuse throughout the brain tissue. Two-photon excitation microscopy confirmed the physical and photometrical stability of these nanoprobes in living zebrafish (Danio rerio) brain tissue, while the addition of Zn2+ quenched the nanoprobe fluorescence. Combining orthogonal sensing methods with our engineered nanoprobes will enable the study of imbalances in homeostatic Zn2+ regulation. The proposed bionanoprobe system offers a versatile platform to couple metal ion specific linkers and contribute to the understanding of neurological diseases.

Impaired Dopamine Release and Latent Learning in Alzheimer's Disease Model Zebrafish.

Alzheimer's disease (AD) is a progressive, fatal, neurodegenerative disorder for which only treatments of limited efficacy are available. Despite early mentions of dementia in the ancient literature and the first patient diagnosed in 1906, the underlying causes of AD are not well understood. This study examined the possible role of dopamine, a neurotransmitter that is involved in cognitive and motor function, in AD. We treated adult zebrafish (Danio rerio) with okadaic acid (OKA) to model AD and assessed the resulting behavioral and neurochemical changes. We then employed a latent learning paradigm to assess cognitive and motor function followed by neurochemical analysis with fast-scan cyclic voltammetry (FSCV) at carbon fiber microelectrodes to measure the electrically stimulated dopamine release. The behavioral assay showed that OKA treatment caused fish to have lower motivation to reach the goal chamber, resulting in impeded learning and decreased locomotor activity compared to controls. Our voltammetric measurements revealed that the peak dopamine overflow in OKA-treated fish was about one-third of that measured in controls. These findings highlight the profound neurochemical changes that may occur in AD. Furthermore, they demonstrate that applying the latent learning paradigm and FSCV to zebrafish is a promising tool for future neurochemical studies and may be useful for screening drugs for the treatment of AD.

Dopamine Release Impairments Accompany Locomotor and Cognitive Deficiencies in Rotenone-Treated Parkinson's Disease Model Zebrafish.

In this work, we carried out neurochemical and behavioral analysis of zebrafish (Danio rerio) treated with rotenone, an agent used to chemically induce a syndrome resembling Parkinson's disease (PD). Dopamine release, measured with fast-scan cyclic voltammetry (FSCV) at carbon-fiber electrodes in acutely harvested whole brains, was about 30% of that found in controls. Uptake, represented by the first order rate constant (k) and the half-life (t1/2) determined by nonlinear regression modeling of the stimulated release plots, was also diminished. Behavioral analysis revealed that rotenone treatment increased the time required for zebrafish to reach a reward within a maze by more than 50% and caused fish to select the wrong pathway, suggesting that latent learning was impaired. Additionally, zebrafish treated with rotenone suffered from diminished locomotor activity, swimming shorter distances with lower mean velocity and acceleration. Thus, the neurochemical and behavioral approaches, as applied, were able to resolve rotenone-induced differences in key parameters. This approach may be effective for screening therapies in this and other models of neurodegeneration.

Graphene-Fiber Microelectrodes for Ultrasensitive Neurochemical Detection.

Here, we have synthesized and characterized graphene-fiber microelectrodes (GFME's) for subsecond detection of neurochemicals with fast-scan cyclic voltammetry (FSCV) for the first time. GFME's exhibited extraordinary properties including faster electron transfer kinetics, significantly improved sensitivity, and ease of tunability that we anticipate will have major impacts on neurochemical detection for years to come. GF's have been used in the literature for various applications; however, scaling their size down to microelectrodes and implementing them as neurochemical microsensors is significantly less developed. The GF's developed in this paper were on average 20-30 µm in diameter and both graphene oxide (GO) and reduced graphene oxide (rGO) fibers were characterized with FSCV. Neat GF's were synthesized using a one-step dimension-confined hydrothermal strategy. FSCV detection has traditionally used carbon-fiber microelectrodes (CFME's) and more recently carbon nanotube fiber electrodes; however, uniform functionalization and direct control of the 3D surface structure of these materials remain limited. The expansion to GFME's will certainly open new avenues for fine-tuning the electrode surface for specific electrochemical detection. When comparing to traditional CFME's, our GFME's exhibited significant increases in electron transfer, redox cycling, fouling resistance, higher sensitivity, and frequency independent behavior which demonstrates their incredible utility as biological sensors.

Optimized Sawhorse Waveform for the Measurement of Oxytocin Release in Zebrafish.

Oxytocin is a nonapeptide hormone involved in numerous physiological functions. Real-time electrochemical measurements of oxytocin in living tissue are challenging due to electrode fouling and the large potentials needed to oxidize the tyrosine residue. Here, we used fast-scan cyclic voltammetry at carbon-fiber microelectrodes and flow injection analysis to optimize a waveform for the measurement of oxytocin. This optimized waveform employed an accumulation potential of -0.6 V, multiple scan rates, and a 3 ms holding potential at a positive, oxidizing potential of +1.4 V before linearly scanning the potential back to -0.6 V (versus Ag/AgCl). We obtained a limit of quantitation of 0.34 ± 0.02 µM, and our electrodes did not foul upon multiple injections. Moreover, to demonstrate the utility of our method, we measured the release of oxytocin, evoked by light application and mechanical perturbation, in whole brains from genetically engineered adult zebrafish that express channelrhodopsin-2 selectively on oxytocinergic neurons. Collectively, this work expands the toolkit for the measurement of peptides in living tissue preparations.

In Situ Electrochemical Monitoring of Caged Compound Photochemistry: An Internal Actinometer for Substrate Release.

Caged compounds are molecules that release a protective substrate to free a biologically active substrate upon treatment with light of sufficient energy and duration. A notable limitation of this approach is difficulty in determining the degree of photoactivation in tissues or opaque solutions because light reaching the desired location is obstructed. Here, we have addressed this issue by developing an in situ electrochemical method in which the amount of caged molecule photorelease is determined by fast-scan cyclic voltammetry (FSCV) at carbon-fiber microelectrodes. Using p-hydroxyphenyl glutamate (pHP-Glu) as our model system, we generated a linear calibration curve for oxidation of 4-hydroxyphenylacetic acid (4HPAA), the group from which the glutamate molecule leaves, up to a concentration of 1000 µM. Moreover, we are able to correct for the presence of residual pHP-Glu in solution as well as the light artifact that is produced. A corrected calibration curve was constructed by photoactivation of pHP-Glu in a 3 µL photoreaction vessel and subsequent analysis by high-performance liquid chromatography. This approach has yielded a linear relationship between 4HPAA concentration and oxidation current, allowing the determination of released glutamate independent of the amount of light reaching the chromophore. Moreover, we have successfully validated the newly developed method by in situ measurement in a whole, intact zebrafish brain. This work demonstrates for the first time the in situ electrochemical monitoring of caged compound photochemistry in brain tissue with FSCV, thus facilitating analyses of neuronal function.

Local and Long-Range Organization in Room Temperature Ionic Liquids.

Room temperature ionic liquids (RTILs) have a wide range of current and potential applications, in areas ranging from supercapacitor energy storage to sequestration of toxic gas phase species and use as reusable solvents for selected organic reactions. All these applications stem from their unique physical and chemical properties, which remain understood to only a limited extent. Among the issues of greatest importance is the extent to which RTILs exist as dissociated ionic species and the length scales over which some types of organizations are seen to exist in them. In this Invited Feature Article, we review the current understanding of organization in this family of materials, where opportunities lie in terms of deepening our understanding, and what potential applications would benefit from gaining such knowledge.

Characterizing the Magnitude and Structure-Dependence of Free Charge Density Gradients in Room-Temperature Ionic Liquids.

We have reported previously on the existence of charge-induced long-range organization in the room-temperature ionic liquid (RTIL), BMIM+BF4-. The induced organization is in the form of a free charge density gradient (ρf) that exists over ca. 100 µm into the RTIL in contact with a charged surface. The fluorescence anisotropy decay of a trace-level charged chromophore in the RTIL is measured as a function of distance from the indium-doped tin oxide support surface to probe this free charge density gradient. We report here on the characterization of the free charge density gradient in five different imidazolium RTILs and use these data to evaluate the magnitude of the induced free charge density gradient. Both the extent and magnitude of this gradient depend on the chemical structures of the cationic and anionic constituents of the RTIL used. Control over the magnitude of ρf has implications for the utility of RTILs for a host of applications that remain to be explored fully.

Inkjet-Printed Carbon Nanotube Electrodes for Measuring Pyocyanin and Uric Acid in a Wound Fluid Simulant and Culture Media.

Polyacrylamide-coated, carbon nanotube (PA/CNT) electrodes were prepared by an inkjet printing process and used to measure pyocyanin and uric acid in a wound fluid simulant at 37 °C. These two molecules are potential indicators of infection, and therefore their detection could prove useful for monitoring wound healing. Pyocyanin is a marker for the common wound bacterium Pseudomonas aeruginosa. Our long-term goal is to use these inexpensive and disposable electrodes to measure biomarkers of wound healing directly. In this proof-of-concept work, studies were performed in a wound fluid simulant to evaluate the stability of the electrodes and their responsiveness for the two bioanalytes. The PA/CNT inkjet-printed electrodes and electrical contacts were stable with unchanging physical and electrochemical properties in the wound fluid simulant over a 7-8-day period at 37 °C. The detection figures of merit for pyocyanin in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 0.10 to 100 µmol L-1 (R2 = 0.9992), limit of detection = 0.10 µmol L-1 (S/N = 3), sensitivity = 35.6 ± 0.8 mA-L mol-1 and response variability ≤4% RSD. The detection figures of merit for uric acid in the simulant at 37 °C were as follows: linear over the physiologically relevant range = 100 to 1000 µmol L-1 (R2 = 0.9997), sensitivity = 2.83 ± 0.01 mA-L mol-1, and response variability ≤4% RSD. The limit of detection was not determined. The PA/CNT electrodes were also used to quantify pyocyanin concentrations in cell-free culture media from different strains of P. aeruginosa. The detected concentrations ranged from 1.00 ± 0.02 to 118 ± 6 µM depending on the strain.

Evaluation of a nitrogen-incorporated tetrahedral amorphous carbon thin film for the detection of tryptophan and tyrosine using flow injection analysis with amperometric detection.

We report on the analytical performance of a tetrahedral amorphous carbon (ta-C:N) thin-film electrode in flow injection analysis with amperometric detection. Two model redox analytes were used to evaluate the electrode response because of their positive detection potentials and propensity (i.e., reaction products) to adsorb and foul sp2 carbon electrodes: tyrosine and tryptophan. ta-C:N electrodes are attractive for electroanalytical applications because they possesses many of the excellent properties of boron-doped nanocrystalline diamond (BDD) and they can be deposited at or near room temperature. The results show that the ta-C:N electrode exhibits lower background current and noise than glassy carbon (GC). The electrode was stable microstructurally at the positive potentials used for detection, ∼1.1 V, of these two amino acids and it exhibited superior analytical detection figures of merit as compared to GC and as good or superior to BDD. The linear dynamic range for both analytes at ta-C:N was from 0.1 to 100 µmol L-1, the sensitivity was 8-12 mA L mol-1, the short-term response variability was 1-2%, and the minimum detectable concentration was 89.7 ± 0.9 nM (18.3 µg L-1 or 0.46 ng) for tryptophan and 120 ± 11 nM (21.7 µg L-1 or 0.54 ng) for tyrosine. The analytical detection figures of merit for these amino acids at GC and BDD are also presented for comparison as is characterization data for the chemical composition and microstructure of the ta-C:N film.

Correction: Evaluation of a nitrogen-incorporated tetrahedral amorphous carbon thin film for the detection of tryptophan and tyrosine using flow injection analysis with amperometric detection.

Correction for 'Evaluation of a nitrogen-incorporated tetrahedral amorphous carbon thin film for the detection of tryptophan and tyrosine using flow injection analysis with amperometric detection' by Romana Jarosová, et al., Analyst, 2016, DOI: .

Charge-Induced Long-Range Order in a Room-Temperature Ionic Liquid.

We report direct evidence for charge-induced long-range (ca. 100 µm) order in the room-temperature ionic liquid (IL) 1-butyl-3-methylimidazolium tetrafluoroborate (BMIM(+)BF4(-)), supported on a silica surface. We have measured the rotational diffusion dynamics of anionic, cationic, and neutral chromophores as a function of distance from a silica surface. The results reflect the excess charge density gradient induced in the IL by the (negative) charge present on the silica surface. Identical measurements in ethylene glycol reveal spatially invariant reorientation dynamics for all chromophores. Capping the silica support with Me2SiCl2 results in spatially invariant reorientation dynamics in the IL. We understand these data in the context of the IL exhibiting a spatially damped piezoelectric response mediated by IL fluidity and disorder.

Structure, Electronic Properties, and Electrochemical Behavior of a Boron-Doped Diamond/Quartz Optically Transparent Electrode.

The morphology, microstructure, chemistry, electronic properties, and electrochemical behavior of a boron-doped nanocrystalline diamond (BDD) thin film grown on quartz were evaluated. Diamond optically transparent electrodes (OTEs) are useful for transmission spectroelectrochemical measurements, offering excellent stability during anodic and cathodic polarization and exposure to a variety of chemical environments. We report on the characterization of a BDD OTE by atomic force microscopy, optical spectroscopy, Raman spectroscopic mapping, alternating-current Hall effect measurements, X-ray photoelectron spectroscopy, and electrochemical methods. The results reported herein provide the first comprehensive study of the relationship between the physical and chemical structure and electronic properties of a diamond OTE and the electrode's electrochemical activity.