Evaluating diverse electrode surface patterns of 3D printed carbon thermoplastic electrochemical sensors.

Electrochemical sensing techniques rely on redox reactions taking place at the electrode surface. The configuration of this surface is of the utmost importance in the advancement of electrochemical sensors. The majority of previous electrode manufacturing methods, including 3D printing have produced electrodes with flat surfaces. There is a distinct potential for 3D printing to create intricate and distinctive electrode surface shapes. In the proposed work, 3D printed carbon black polylactic acid electrodes with nine different surface morphologies were made. These were compared to a flat surface electrode. To evaluate the performance of the electrodes, measurements were conducted in three different redox probes (ferrocene methanol, ferricyanide, and dopamine). Our findings highlighted that when electrodes were normalised for the geometric surface area of the electrode, the surface pattern of the electrode surface can impact the observed current and electron transfer kinetics. Electrodes that had a dome and flag pattern on the electrode surface showed the highest oxidation currents and had lower values for the difference between the anodic and cathodic peak current (ΔE). However, designs with rings had lower current values and higher ΔE values. These differences are most likely due to variations in the accessibility of conductive sites on the electrode surface due to the varying surface roughness of different patterned designs. Our findings highlight that when making electrodes using 3D printing, surface patterning of the electrode surface can be used as an effective approach to enhance the performance of the sensor for varying applications.

Monitoring Gut Epithelium Serotonin and Melatonin Overflow Provides Spatial Mapping of Inflammation.

Electrochemical arrays were used to measure the overflow of serotonin (5-HT) and melatonin (MEL) from the entire colon of healthy mice and mice with chemical-induced inflammatory bowel disease (IBD), to understand the interplay between inflammation and colonic function. We show that 5-HT overflow is increased, whilst MEL levels are reduced, in inflamed tissues. The levels of MEL are increased at the interface between healthy and inflamed regions within the colon and may limit the spread of inflammation. Understanding the interplay between inflammation and mucosal epithelial signalling can provide key insight into colonic function and aid the development of effective therapeutic strategies to treat gastrointestinal diseases.

Infection responsive coatings to reduce biofilm formation and encrustation of urinary catheters.

AIMS: The care of patients undergoing long-term urethral catheterization is frequently complicated by Proteus mirabilis infection. This organism forms dense, crystalline biofilms, which block catheters leading to serious clinical conditions. However, there are currently no truly effective approaches to control this problem. Here, we describe the development of a novel theranostic catheter coating, to simultaneously provide early warning of blockage, and actively delay crystalline biofilm formation. METHODS AND RESULTS: The coating comprises of a pH sensitive upper polymer layer (poly(methyl methacrylate-co-methacrylic acid); Eudragit S 100®) and a hydrogel base layer of poly(vinyl alcohol), which is loaded with therapeutic agents (acetohydroxamic acid or ciprofloxacin hydrochloride) and a fluorescent dye, 5(6)-carboxyfluorescein (CF). The elevation of urinary pH due to P. mirabilis urease activity results in the dissolution of the upper layer and release of cargo agents contained in the base layer. Experiments using in vitro models, which were representative of P. mirabilis catheter-associated urinary tract infections, demonstrated that these coatings significantly delay time taken for catheters to block. Coatings containing both CF dye and ciprofloxacin HCl were able to provide an average of ca. 79 h advanced warning of blockage and extend catheter lifespan ca. 3.40-fold. CONCLUSIONS: This study has demonstrated the potential for theranostic, infection-responsive coatings to form a promising approach to combat catheter encrustation and actively delay blockage.

Decreased 14-3-3 expression correlates with age-related regional reductions in CNS dopamine and motor function in the pond snail, Lymnaea.

Ageing is associated in many organisms with a reduction in motor movements. We have previously shown that the rate of feeding movements of the pond snail, Lymnaea, decreased with age but the underlying cause is not fully understood. Here, we show that dopamine in the cerebro-buccal complex is an important signalling molecule regulating feeding frequency in Lymnaea and that ageing is associated with a decrease in CNS dopamine. A proteomic screen of young and old CNSs highlighted a group of proteins that regulate stress responses. One of the proteins identified was 14-3-3, which can enhance the synthesis of dopamine. We show that the Lymnaea 14-3-3 family exists as three distinct isoforms. The expression of the 29 kDa isoform (14-3-3Lym3) in the cerebro-buccal complex decreased with age and correlated with feeding rate. Using a 14-3-3 antagonist (R18) we were able to reduce the synthesis of L-DOPA and dopamine in ex vivo cerebro-buccal complexes. Together these data suggest that an age-related reduction in 14-3-3 can decrease CNS dopamine leading to a consequential reduction in feeding rate.

Nano-analysis Reveals High Fraction of Serotonin Release during Exocytosis from a Gut Epithelium Model Cell.

Electrochemical methods were used to explore the exocytotic nature of serotonin (5-HT) release in human carcinoid BON cells, an in vitro human enterochromaffin cell model, to understand the mechanisms operating the release of gut-derived 5-HT in the intestinal mucosal epithelium. We show that the fractional vesicular 5-HT release in BON cells is 80 % compared to previous work in pancreatic beta cells (34 %). The fractional release increased from 80 % in control BON cells to 87 % with 5-HT preincubation and nearly 100 % with the combination of 5-HT and the 5-HT4 autoreceptor agonist, cisapride. Thus, partial release is the primary mechanism of exocytosis in BON cells, resulting in a variable amount of the vesicular content being released. Factors that control secretion of 5-HT from enterochromaffin cells or BON cells are important as partial release provides a mechanism for development of effective therapeutic strategies to treat gastrointestinal diseases.

Recycling Chocolate Aluminum Wrapping Foil as to Create Electrochemical Metal Strip Electrodes.

The development of low-cost electrode devices from conductive materials has recently attracted considerable attention as a sustainable means to replace the existing commercially available electrodes. In this study, two different electrode surfaces (surfaces 1 and 2, denoted as S1 and S2) were fabricated from chocolate wrapping aluminum foils. Energy dispersive X-Ray (EDX) and field emission scanning electron microscopy (FESEM) were used to investigate the elemental composition and surface morphology of the prepared electrodes. Meanwhile, cyclic voltammetry (CV), chronoamperometry, electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) were used to assess the electrical conductivities and the electrochemical activities of the prepared electrodes. It was found that the fabricated electrode strips, particularly the S1 electrode, showed good electrochemical responses and conductivity properties in phosphate buffer (PB) solutions. Interestingly, both of the electrodes can respond to the ruthenium hexamine (Ruhex) redox species. The fundamental results presented from this study indicate that this electrode material can be an inexpensive alternative for the electrode substrate. Overall, our findings indicate that electrodes made from chocolate wrapping materials have promise as electrochemical sensors and can be utilized in various applications.

Determination of tryptophan metabolism from biological tissues and fluids using high performance liquid chromatography with simultaneous dual electrochemical detection.

Serotonin and kynurenine are formed following metabolism of the essential amino acid tryptophan. Both molecules play important biological roles and the balance of how tryptophan metabolism varies to either the serotonin or kynurenine pathway may provide key insight into the inflammatory status of the biological region. At present complex chromatographic methods are utilised which predominately focus on either monitoring analytes in the serotonin or kynurenine pathway rather than both. Our study develops a simple yet robust methodology for the monitoring of tryptophan metabolism. We utilised isocratic reverse phase high-performance liquid chromatography with simultaneously dual electrochemical detection. This approach allowed for separation of co-eluted analytes and identification of analytes from both pathways within 14 minutes. For all analytes, limits of detection were <35 nM. No crosstalk was observed when dual simultaneous detection was conducted in a radial flow cell. Responses from the hippocampus, blood and ileum mucosa highlighted that each region had a varying ratio of serotonin to kynurenine pathway, indicating varied approaches to tryptophan metabolism. The developed method can monitor how the metabolism of tryptophan varies between the two pathways which can provide insight into the inflammatory state of reach region with age and disease.

3D Printed Molds Encompassing Carbon Composite Electrodes To Conduct Multisite Monitoring in the Entire Colon.

The activity of the colon is regulated by chemical signaling, of which serotonin (5-HT) is a key transmitter. Monitoring of mucosal 5-HT overflow has been achieved to date using microelectrodes on a small segment of colonic tissue; however, little is known if such measurements are reflective with regards to 5-HT signaling from the entire colon. This study focused on developing an electrochemical array device that could be utilized to conduct multisite measurements of 5-HT overflow from the entire colon. A 3D printed mold was fabricated that could house 6 multiwall carbon nanotube composite electrodes and provide a fixed distance between the electrodes and the tissue along the entire length of the colon. The electrodes were assessed for sensitivity, stability, and crosstalk before conducting in vitro measurements using colons obtained from 6- and 24-month old mice. As composite electrodes can have a high degree of variability, normalization factors were required between electrodes for a given array. The device had the sensitivity and stability required for 5-HT measurements from intestinal tissue. Regio-specific changes in 5-HT overflow were observed with age, where increases in 5-HT overflow were observed in the distal colon due to an impairment/loss in the serotonin transporter (SERT). Our strategy can be utilized to develop arrays of varying sizes and geometries, which can offer practical solutions for large-scale tissue measurements.

Lithium Ion Sensors.

The detection and monitoring of lithium in environmental and clinical settings is becoming increasingly important. In this review, sensors incorporating conductive polymers and lithium bronzes are discussed, together with electrochemical and spectroscopic approaches. Ionophore-based methods have been employed extensively, with varying degrees of selectivity and sensitivity, and these are discussed in depth.

Real-time detection of carboplatin using a microfluidic system.

A microfluidic sensor system based on a carbon nanotube-epoxy composite electrode was fabricated to allow detection of the presence of the anti-cancer drug carboplatin in healthy tissue in real time during chemotherapy. Detection of carboplatin was carried out by observing the effects of the drug on the differential pulse voltammetry of free purine bases using a novel carbon nanotube-epoxy composite electrode. In free solution these electrodes performed better than glassy carbon electrodes for oxidation of the free purine bases AMP and GMP, and than DNA-modified carbon nanotube-epoxy composite sensors for detection of carboplatin. On-line carboplatin detection was performed using a computer-controlled microfluidic platform. The methodology for on-line carboplatin detection was optimised in terms of the analysis time and to allow repeated carboplatin measurement using the same electrode. Microdialysis sampling and our microfluidic platform were combined to give a proof-of-concept system for real-time carboplatin detection with a limit of detection of 0.014 µM carboplatin in the sampled media. This paper is dedicated to Craig Lunte's pioneering work in analysis and microdialysis.

Amperometry approach curve profiling to understand the regulatory mechanisms governing the concentration of intestinal extracellular serotonin.

Enterochromaffin (EC) cells located within the intestinal mucosal epithelium release serotonin (5-HT) to regulate motility tones, barrier function and the immune system. Electroanalytical methodologies have been able to monitor steady state basal extracellular 5-HT levels but are unable to provide insight into how these levels are influenced by key regulatory processes such as release and uptake. We established a new measurement approach, amperometry approach curve profiling, which monitors the extracellular 5-HT level at different electrode-tissue (E-T) distances. Analysis of the current profile can provide information on contributions of regulatory components on the observed extracellular 5-HT level. Measurements were conducted from ex vivo murine ileum and colon using a boron-doped diamond (BDD) microelectrode. Amperometry approach curve profiling coupled with classical pharmacology demonstrated that extracellular 5-HT levels were significantly lower in the colon when compared to the ileum. This difference was due to a greater degree of activity of the 5-HT transporter (SERT) and a reduced amount of 5-HT released from colonic EC cells. The presence of an inhibitory 5-HT4 autoreceptor was observed in the colon, where a 40% increase in extracellular 5-HT was the half maximal inhibitory concentration for activation of the autoreceptor. This novel electroanalytical approach allows estimates of release and re-uptake and their contribution to 5-HT extracellular concentration from intestinal tissue be obtained from a single series of measurements.

Monitoring the electroactive cargo of extracellular vesicles can differentiate various cancer cell lines.

Extracellular vesicles (EVs) are pivotal in cell-to-cell communication due to the array of cargo contained within these vesicles. EVs are considered important biomarkers for identification of disease, however most measurement approaches have focused on monitoring specific surface macromolecular targets. Our study focuses on exploring the electroactive component present within cargo from EVs obtained from various cancer and non-cancer cell lines using a disk carbon fiber microelectrode. Variations in the presence of oxidizable components were observed when the total cargo from EVs were measured, with the highest current detected in EVs from MCF7 cells. There were differences observed in the types of oxidizable species present within EVs from MCF7 and A549 cells. Single entity measurements showed clear spikes due to the detection of oxidizable cargo within EVs from MCF7 and A549 cells. These studies highlight the promise of monitoring EVs through the presence of varying electroactive components within the cargo and can drive a wave of new strategies towards specific detection of EVs for diagnosis and prognosis of various diseases.

Pristine Ti3C2Tx MXene Enables Flexible and Transparent Electrochemical Sensors.

In the era of the internet of things, there exists a pressing need for technologies that meet the stringent demands of wearable, self-powered, and seamlessly integrated devices. Current approaches to developing MXene-based electrochemical sensors involve either rigid or opaque components, limiting their use in niche applications. This study investigates the potential of pristine Ti3C2Tx electrodes for flexible and transparent electrochemical sensing, achieved through an exploration of how material characteristics (flake size, flake orientation, film geometry, and uniformity) impact the electrochemical activity of the outer sphere redox probe ruthenium hexamine using cyclic voltammetry. The optimized electrode made of stacked large Ti3C2Tx flakes demonstrated excellent reproducibility and resistance to bending conditions, suggesting their use for reliable, robust, and flexible sensors. Reducing electrode thickness resulted in an amplified faradaic-to-capacitance signal, which is advantageous for this application. This led to the deposition of transparent thin Ti3C2Tx films, which maintained their best performance up to 73% transparency. These findings underscore its promise for high-performance, tailored sensors, marking a significant stride in advancing MXene utilization in next-generation electrochemical sensing technologies. The results encourage the analytical electrochemistry field to take advantage of the unique properties that pristine Ti3C2Tx electrodes can provide in sensing through more parametric studies.

A planar microelectrode array for simultaneous detection of electrically evoked dopamine release from distinct locations of a single isolated neuron.

Neurotransmission is a key process of communication between neurons. Although much is known about this process and the influence it has on the function of the body, little is understood about the dynamics of signalling from structural regions of a single neuron. In this study we have fabricated and characterised a microelectrode array (MEA) which was utilised for simultaneous multi-site recordings of dopamine release from an isolated single neuron. The MEA consisted of gold electrodes that were created in plane with the insulation layer using a chemical mechanical planarization process. The detection limit for dopamine measurements was 11 ± 3 nM and all the gold electrodes performed in a consistent fashion during amperometric recordings of 100 nM dopamine. Fouling of the gold electrode was investigated, where no significant change in the current was observed over 4 hours when monitoring 100 nM dopamine. The MEA was accessed using freshly isolated dopaminergic somas from the pond snail, Lymnaea stagnalis, where electrically evoked dopamine release was clearly observed. Measurements were conducted at four structural locations of a single isolated neuron, where electrically evoked dopamine release was observed from the cell body, axonal regions and the terminal. Over time, the release of dopamine varied over the structural regions of the neuron. Such information can provide an insight into the signalling mechanism of neurons and how they potentially form synaptic connections.

Influence of instrument parameters on the electrochemical activity of 3D printed carbon thermoplastic electrodes.

3D printing provides a reliable approach for the manufacture of carbon thermoplastic composite electrochemical sensors. Many studies have explored the impact of printing parameters on the electrochemical activity of carbon thermoplastic electrodes but limited is known about the influence of instrument parameters, which have been shown to alter the structure and mechanical strength of 3D printed thermoplastics. We explored the impact of extruder temperature, nozzle diameter and heated bed temperature on the electrochemical activity of carbon black/poly-lactic acid (CB/PLA) electrodes. Cyclic voltammetry and electrochemical impedance spectroscopy measurements were conducted using standard redox probes. The electrode surface and cross-section of the electrode was visualised using scanning electron microscopy. We found that using extruder temperatures of 230 °C and 240 °C improved the electrochemical activity of CB/PLA electrodes, due to an increase in surface roughness and a reduction in the number of voids in-between print layers. Nozzle diameter, heated bed temperature of different 3D printers did not impact the electrochemical activity of CB/PLA electrodes. However high-end printers provide improved batch reproducibility of electrodes. These findings highlight the key instrument parameters that need to be considered when manufacturing carbon thermoplastic composite electrochemical sensors when using 3D printing.

Three-Dimensional-Printed Electrochemical Multiwell Plates for Monitoring Food Intolerance from Intestinal Organoids.

Common symptoms of food intolerance are caused by chemical components within food that have a pharmacological activity to alter the motility of the gastrointestinal tract. Food intolerance is difficult to diagnose as it requires a long-term process of eliminating foods that are responsible for gastrointestinal symptoms. Enterochromaffin (EC) cells are key intestinal epithelium cells that respond to luminal chemical stimulants by releasing 5-HT. Changes in 5-HT levels have been shown to directly alter the motility of the intestinal tract. Therefore, a rapid approach for monitoring the impact of chemicals in food components on 5-HT levels can provide a personalized insight into food intolerance and help stratify diets. Within this study, we developed a three-dimensional (3D)-printed electrochemical multiwell plate to determine changes in 5-HT levels from intestinal organoids that were exposed to varying chemical components found in food. The carbon black/poly-lactic acid (CB/PLA) electrodes had a linear range in physiological concentrations of 5-HT (0.1-2 µM) with a limit of detection of 0.07 µM. The electrodes were stable for monitoring 5-HT overflow from intestinal organoids. Using the electrochemical multiwell plate containing intestinal organoids, increases in 5-HT were observed in the presence of 0.1 mM cinnamaldehyde and 10 mM quercetin but reduction in 5-HT levels was observed in 1 mM sorbitol when compared to control. These changes in the presence of chemicals commonly found in food were verified with ex vivo ileum tissue measurements using chromatography and amperometry with boron-doped diamond electrodes. Overall, our 3D electrochemical multiwell plate measurements with intestinal organoids highlight an approach that can be a high-throughput platform technology for rapid screening of food intolerance to provide personalized nutritional diet.

Simultaneous detection of gastric acid and histamine release to unravel the regulation of acid secretion from the guinea pig stomach.

Gastric acid secretion is regulated by three primary components that activate the parietal cell: histamine, gastrin, and acetylcholine (ACh). Although much is known about these regulatory components individually, little is known on the interplay of these multiple activators and the degree of regulation they pose on the gastric acid secretion mechanism. We utilized a novel dual-sensing approach, where an iridium oxide sensor was used to monitor pH and a boron-doped diamond electrode was used for the detection of histamine from in vitro guinea pig stomach mucosal sections. Under basal conditions, gastrin was shown to be the main regulatory component of the total acid secretion and directly activated the parietal cell rather than by mediating gastric acid secretion through the release of histamine from the enterochromaffin-like cell, although both pathways were active. Under stimulated conditions with ACh, the gastrin and histamine components of the total acid secretion were not altered compared with levels observed under basal conditions, suggestive that ACh had no direct effect on the enterochromaffin-like cell and G cell. These data identify a new unique approach to investigate the regulation pathways active during acid secretion and the degree that they are utilized to drive total gastric acid secretion. The findings of this study will enhance our understanding on how these signaling mechanisms vary under pathophysiology or therapeutic management.

Chromatographic assay to study the activity of multiple enzymes involved in the synthesis and metabolism of dopamine and serotonin.

Serotonin and dopamine are crucial regulators of signalling in the peripheral and central nervous systems. We present an ex-vivo, isocratic chromatographic method that allows for the measurement of tyrosine, L-3,4-dihydroxyphenylalanine (L-DOPA), dopamine, 3,4-dihydroxyphenylacetic acid (DOPAC), tryptophan, 5-hydroxytryptophan (5-HTP), serotonin and 5-hydroxy-3-indoleacetic acid (5-HIAA) in a model central nervous (CNS) system, to study the role of key enzymes involved in the synthesis and metabolism of serotonin and dopamine. By utilising a sample splitting technique, we could test a single CNS sample at multiple time points under various pharmacological treatments. In, addition, we were able to conduct this assay by utilising the endogenous biochemical components of the CNS to study the synthesis and metabolism of serotonin and dopamine, negating the requirement of additional enzyme activators or stabilisers in the biological matrix. Finally we utilised NSD-1015, an aromatic amino acid decarboxylase enzyme inhibitor used to study the synthesis of dopamine and serotonin to monitor alterations in levels of key neurochemicals. 3-hydroxybenzylhydrazine dihydrochloride (NSD-1015) was able to reduce levels of serotonin and dopamine, whilst elevating precursors L-DOPA and 5-HTP.