Somatostatin modulation of initial fusion pores in Ca2+-triggered exocytosis from mouse chromaffin cells.

Somatostatin, a peptide hormone that activates G-protein-coupled receptors, inhibits the secretion of many hormones. This study investigated the mechanisms of this inhibition using amperometry recording of Ca2+-triggered catecholamine secretion from mouse chromaffin cells. Two distinct stimulation protocols, high-KCl depolarization and caffeine, were used to trigger exocytosis, and confocal fluorescence imaging was used to monitor the rise in intracellular free Ca2+. Analysis of single-vesicle fusion events (spikes) resolved the action of somatostatin on fusion pores at different stages. Somatostatin reduced spike frequency, and this reduction was accompanied by prolongation of pre-spike feet and slowing of spike rise times. This indicates that somatostatin stabilizes initial fusion pores and slows their expansion. This action on the initial fusion pore impacted the release mode to favour kiss-and-run over full-fusion. During a spike the permeability of a fusion pore peaks, declines and then settles into a plateau. Somatostatin had no effect on the plateau, suggesting no influence on late-stage fusion pores. These actions of somatostatin were indistinguishable between exocytosis triggered by high-KCl and caffeine, and fluorescence imaging showed that somatostatin had no effect on stimulus-induced rises in cytosolic Ca2+. Our findings thus demonstrate that the signalling cascades activated by somatostatin target the exocytotic machinery that controls the initial and expanding stages of fusion pores, while having no effect on late-stage fusion pores. As a result of its stronger inhibition of full-fusion compared to kiss-and-run, somatostatin will preferentially inhibit the secretion of large peptides over the secretion of small catecholamines. KEY POINTS: Somatostatin inhibits the secretion of various hormones by activating G-protein-coupled receptors. In this study, we used amperometry to investigate the mechanism by which somatostatin inhibits catecholamine release from mouse chromaffin cells. Somatostatin increased pre-spike foot lifetime and slowed fusion pore expansion. Somatostatin inhibited full-fusion more strongly than kiss-and-run. Our results suggest that the initial fusion pore is the target of somatostatin-mediated regulation of hormone release. The stronger inhibition of full-fusion by somatostatin will result in preferential inhibition of peptide release.

Dynamin-2 mutations linked to neonatal-onset centronuclear myopathy impair exocytosis and endocytosis in adrenal chromaffin cells.

Dynamins are large GTPases whose primary function is not only to catalyze membrane scission during endocytosis but also to modulate other cellular processes, such as actin polymerization and vesicle trafficking. Recently, we reported that centronuclear myopathy associated dynamin-2 mutations, p.A618T, and p.S619L, impair Ca2+-induced exocytosis of the glucose transporter GLUT4 containing vesicles in immortalized human myoblasts. As exocytosis and endocytosis occur within rapid timescales, here we applied high-temporal resolution techniques, such as patch-clamp capacitance measurements and carbon-fiber amperometry to assess the effects of these mutations on these two cellular processes, using bovine chromaffin cells as a study model. We found that the expression of any of these dynamin-2 mutants inhibits a dynamin and F-actin-dependent form of fast endocytosis triggered by single action potential stimulus, as well as inhibits a slow compensatory endocytosis induced by 500 ms square depolarization. Both dynamin-2 mutants further reduced the exocytosis induced by 500 ms depolarizations, and the frequency of release events and the recruitment of neuropeptide Y (NPY)-labeled vesicles to the cell cortex after stimulation of nicotinic acetylcholine receptors with 1,1-dimethyl-4-phenyl piperazine iodide (DMPP). They also provoked a significant decrease in the Ca2+-induced formation of new actin filaments in permeabilized chromaffin cells. In summary, our results indicate that the centronuclear myopathy (CNM)-linked p.A618T and p.S619L mutations in dynamin-2 affect exocytosis and endocytosis, being the disruption of F-actin dynamics a possible explanation for these results. These impaired cellular processes might underlie the pathogenic mechanisms associated with these mutations.

Disposable biosensor based on ionic liquid, carbon nanofiber and poly(glutamic acid) for tyramine determination.

In this study, an electrochemical biosensor based on carbon nanofibers (CNF), ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate (IL), poly(glutamic acid) (PGA) and tyrosinase (Tyr) modified screen printed carbon electrode (SPE) was constructed for tyramine determination. Optimum experimental parameters such as CNF and IL amount, polymerization conditions of glutamic acid, enzyme loading, pH of test solution and operating potential were explored. The construction steps of the Tyr/PGA/CNF-IL/SPE were pursued by scanning electron microscopy and cyclic voltammetry. The Tyr/PGA/CNF-IL/SPE biosensor exhibited linear response to tyramine in the range of 2.0 × 10-7 - 4.8 × 10-5 M with a low detection limit of 9.1 × 10-8 M and sensitivity of 302.6 µA mM-1. The other advantages of Tyr/PGA/CNF-IL/SPE include its high reproducibility, good stability and anti-interference ability. The presented biosensor was also applied for tyramine determination in malt drink and pickle juice samples and mean analytical recoveries of spiked tyramine were calculated as 100.6% and 100.4% respectively.

Novel MoS2-decorated Cu2O hybrid nanoparticles for enhanced non-enzymatic electrochemical cholesterol detection.

Precise identification of cholesterol levels is crucial for the early diagnosis of cardiovascular risk factors. This paper presents a novel approach for cholesterol detection that circumvents the reliance on enzymatic processes. Leveraging the unique properties of advanced materials and electrochemical principles, our non-enzymatic approach demonstrates enhanced sensitivity, specificity, and limit of detection in cholesterol analysis. A non-enzymatic electrochemical biosensor for Cholesterol, employing a nanohybrid comprising Cu2O nanoparticles decorated with MoS2, is presented. The cyclic voltammetry (CV), differential pulse voltammetry (DPV), and amperometry techniques were employed to investigate the electrochemical behaviour of the glassy carbon electrode modified with the Cu2O/MoS2nanohybrid. The modified electrode exhibited an excellent sensitivity of 111.74µAµM-1cm-2through the CV method and showcased a low detection limit of 2.18µM and an expansive linear range spanning 0.1-180µM when employing the DPV method. The electrode also showed good selectivity to various interfering components in 0.1 M NaOH and a satisfied stability of about 15 days at room temperature. The study demonstrates the potential for broader applications in clinical diagnostics and monitoring cardiovascular health, paving the way for a paradigm shift in cholesterol detection methodologies and offering a more efficient and cost-effective alternative to traditional enzymatic assays.

Electrochemiluminescent imaging of a NADH-based enzymatic reaction confined within giant liposomes.

Herein, transient releases either from NADH-loaded liposomes or enzymatic reactions confined in giant liposomes were imaged by electrochemiluminescence (ECL). NADH was first encapsulated with the [Ru(bpy)3]2+ luminophore inside giant liposomes (around 100 µm in diameter) made of DOPC/DOPG phospholipids (i.e., 1,2-dioleolyl-sn-glycero-3-phosphocholine/1,2-dioleoyl-sn-glycerol-3-phospho-(1'-rac-glycerol) sodium salt) on their inner- and outer-leaflet, respectively. Then, membrane permeabilization triggered upon contact between the liposome and a polarized ITO electrode surface and ECL was locally generated. Combination of amperometry, photoluminescence, and ECL provided a comprehensive monitoring of a single liposome opening and content release. In a second part, the work is focused on the ECL characterization of NADH produced by glucose dehydrogenase (GDH)-catalyzed oxidation of glucose in the confined environment delimited by the liposome membrane. This was achieved by encapsulating both the ECL and catalytic reagents (i.e., the GDH, glucose, NAD+, and [Ru(bpy)3]2+) in the liposome. In accordance with the results obtained, NADH can be used as a biologically compatible ECL co-reactant to image membrane permeabilization events of giant liposomes. Under these conditions, the ECL signal duration was rather long (around 10 s). Since many enzymatic reactions involve the NADH/NAD+ redox couple, this work opens up interesting prospects for the characterization of enzymatic reactions taking place notably in artificial cells and in confined environments.

Point-of-care testing device platform for the determination of creatinine on an enzyme@CS/PB/MXene@AuNP-based screen-printed carbon electrode.

Screen-printed carbon electrodes (SPCE) functionalized with MXene-based three-dimensional nanomaterials are reported for rapid determination of creatinine. Ti3C2TX MXene with in situ reduced AuNPs (MXene@AuNP) were used as a coreactant accelerator for efficient immobilization of enzymes. Creatinine could be oxidized by chitosan-embedded creatinine amidohydrolase, creatine amidinohydrolase, or sarcosine oxidase to generate H2O2, which could be electrochemically detected enhanced by Prussian blue (PB). The enzyme@CS/PB/MXene@AuNP/SPCE detected creatinine within the range 0.03-4.0 mM, with a limit of detection of 0.01 mM, with an average recovery of 96.8-103.7%. This indicates that the proposed biosensor is capable of detecting creatinine in a short amount of time (4 min) within a ± 5% percentage error, in contrast with the standard clinical colorimetric method. With this approach, reproducible and stable electrochemical responses could be achieved for determination of creatinine in serum, urine, or saliva. These results demonstrated its potential for deployment in resource-limited settings for early diagnosis and tracking the progression of chronic kidney disease (CKD).

Interfacial galvanic replacement strategy for Pd-doped NiFe MOF nanosheets with highly efficient dopamine detection.

An interfacial galvanic replacement strategy to controllable synthesize palladium nanoparticles (Pd NPs)-modified NiFe MOF nanocomposite on nickel foam, which served as an efficient sensing platform for quantitative determination of dopamine (DA). Pd NPs grown in situ on the nanosheets of NiFe MOF via self-driven galvanic replacement reaction (GRR) and well uniform distribution was achieved. This method effectively reduced the aggregation of metallic nanoparticles and significantly promoted the electron transfer rate during the electrochemical process, leading to improved electrocatalytic activity for DA oxidation. Remarkably, the precisely constructed biosensor achieved a low detection limit (LOD) of 0.068 µM and recovery of 94.1% (RSD 6.7%, N = 3) for simulated real sample detection and also exhibited superior selectivity and stability. The results confirmed that the as-fabricated Pd-NiFe/NF composite electrode could realize the quantitative determination of DA and showed promising prospects in real sample biosensing.

Dual single atomic Ni sites constructing Janus hollow graphene for boosting electrochemical sensing of glucose.

Single atom catalysts (SACs) have attracted attention due to their excellent catalysis activity under specific reactions and conditions. However, the low density of SACs greatly limits catalytic performance. The three-dimensional graphene hollow nanospheres (GHSs) with very thin shell structure can be used as excellent carrier materials. Not only can its outer surface be used to anchor metal single atoms, but its inner surface can also provide rich sites. Here, a novel step-by-step assembly strategy is reported to anchor nickel single atoms (Ni SAs) on the inner and outer surfaces of GHSs (Ni SAs/GHSs/Ni SAs), which significantly increases the loading capacity of Ni SAs (4.8 wt%). Compared to conventional materials that only anchor Ni SAs to the outer surface of the carrier (Ni SAs/GHSs), Ni SAs/GHSs/Ni SAs exhibits significantly higher electrocatalytic activity toward glucose oxidation in alkaline media. The sensitivity of Ni SAs/GHSs/Ni SAs/GCE is nearly five times higher than that of Ni SAs/GHSs/GCE. Moreover, the sensor based on Ni SAs/GHSs/Ni SAs can detect glucose in a wide concentration range of 0.8 µM-1.1244 mM with a low detection limit of 0.19 µM (S/N = 3). This study not only provides an effective sensing material for glucose detection, but also opens a new avenue to construct high-density metal SACs.

CuxO decorated Ti3C2Tx MXene composites for non-enzymatic glucose sensing with large linear ranges.

Ti3C2Tx MXene/CuxO composites were prepared by acid etching combined with electrochemical technique. The abundant active sites on the surface of MXene greatly increase the loading of CuxO nanoparticles, and the synergistic effect between the different components of the composite can accelerate the oxidation reaction of glucose. The results indicate that at the working potential of 0.55 V (vs. Ag/AgCl), the glucose sensor based on Ti3C2Tx MXene/CuxO composite presents large linear concentration ranges from 1 µM to 4.655 mM (sensitivity of 361 µA mM-1 cm-2) and from 5.155 mM to 16.155 mM (sensitivity of 133 µA mM-1 cm-2). The limit of detection is 0.065 µM. In addition, the sensor effectively avoids the oxidative interference of common interfering species such as ascorbic acid, dopamine and uric acid. The sensor has good reproducibility, stability and acceptable recoveries for the detection of glucose in human sweat sample (97.5-103.3%) with RSD values less than 4%. Based on these excellent properties it has great potential for the detection of glucose in real samples.

Eco-friendly spark-generated CoxOy nanoparticle-modified graphite screen-printed sensing surfaces for the determination of H2O2 in energy drinks.

The modification of graphite screen-printed electrodes (SPEs) is reported using an eco-friendly and extremely fast method based on the direct cobalt pin electrode-to-SPE spark discharge at ambient conditions. This approach does not utilize any liquids or chemical templates, does not produce any waste, and allows the in-situ generation of CoxOy nanoparticles onto the electrode surface and the development of efficient electrocatalytic sensing surfaces for the determination of H2O2. Co-spark SPEs were characterized using scanning electron microscopy, energy-dispersive X-ray spectroscopy and x-ray photoelectron spectroscopy (XPS), revealing the formation of surface confined CoxOy nanoparticles and the diverse oxidation states of cobalt species. Co-spark SPEs were also characterized with cyclic voltammetry and electrochemical impedance spectroscopy. Redox transitions of the surface confined electrocatalysts are demonstrated by electrochemical polarization studies, showing the formation of different oxides (CoxOy), varying the XPS results. Amperometric measurements at 0.3 V vs. Ag/AgCl revealed a linear relationship between the current response and the concentration of H2O2 over the range 1 - 102 µM, achieving a limit of detection (3σ/m) of 0.6 µM. The interference effect of various electroactive species was effectively addressed by employing dual measurements in the absence and presence of the enzyme catalase. The analytical utility of the method was evaluated in antioxidant rich real-world samples, such as energy drinks, demonstrating sufficient recovery.

An enzyme-free sensor based on La-doped CoFe-layered double hydroxide decorated on reduced graphene oxide for sensitive electrochemical detection of urea.

The successful synthesis of La-doped CoFe LDH@rGO nanocomposite is reported combining the advantages of LDH and rGO and shows promising performances in electrochemical sensors. The structure of the obtained nanocomposite was investigated using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction pattern (XRD), and field emission scanning electron microscope images (FE-SEM). Then, it was directly utilized to construct a carbon paste electrode (CPE) for urea detection. The electrochemical performance of the sensor was evaluated by various electrochemical methods. The La-CoFe LDH@rGO electrode exhibited excellent electrocatalytic properties, including a wide linear working range of 0.001-23.5 mM, very high sensitivity of 1.07 ± 0.023 µA µM-1 cm-2, a low detection limit of 0.33 ± 0.11 µM, and rapid response time of 5 s towards urea detection at the working potential of 0.4 V. Furthermore, the sensor displayed a high selectivity in different matrices, appropriate reproducibility, and long shelf life without activity loss during 3 months of storage under ambient conditions. Further tests were performed on serum and milk samples to confirm the capability of the proposed sensor for practical applications, demonstrating a reasonable recovery of 94.8 to 102% with an RSD value below 3%. Consequently, the synergistic effect of each component led to the good electrocatalytic activity of the modified electrode towards urea.

Contributing to the management of viral infections through simple immunosensing of the arachidonic acid serum level.

A trendsetting direct competitive-based biosensing tool has been developed and implemented for the determination of the polyunsaturated fatty acid arachidonic acid (ARA), a highly significant biological regulator with decisive roles in viral infections. The designed methodology involves a competitive reaction between the target endogenous ARA and a biotin-ARA competitor for the recognition sites of anti-ARA antibodies covalently attached to the surface of carboxylic acid-coated magnetic microbeads (HOOC-MµBs), followed by the enzymatic label of the biotin-ARA residues with streptavidin-horseradish peroxidase (Strep-HRP) conjugate. The resulting bioconjugates were magnetically trapped onto the sensing surface of disposable screen-printed carbon transducers (SPCEs) to monitor the extent of the biorecognition reaction through amperometry. The operational functioning of the exhaustively optimized and characterized immunosensing bioplatform was highly convenient for the quantitative determination of ARA in serum samples from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2-) and respiratory syncytial virus (RSV)-infected individuals in a rapid, affordable, trustful, and sensitive manner.

2D/1D VSe2/MWCNT hybrid-based electrochemical sensor for carbendazim quantification of environmental, food, and biological samples.

For the first time the sensitive determination of carbendatim (CRB) is reported utilizing a well-designed sensing architecture based on vanadium diselenide-multiwalled carbon nanotube (VSMC). FTIR, XRD, FESEM, EDS, and EIS were employed to evaluate the sensor's structural integrity, and the results demonstrated the successful integration of nanomaterials, resulting in a robust and sensitive electrochemical sensor. Cyclic voltammetry (CV) and chronoamperometric (CA) investigations showed that the sensor best performed at pH 8.0 (BRB) with an excellent detection limit of 9.80 nM with a wide linear range of 0.1 to 10.0 µM. A more thermodynamically viable oxidation of CRB was observed at the VSMC/GCE, with a shift of 200 mV in peak potential towards the less positive side compared with the unmodified GCE. In addition, the sensor demonstrated facile heterogeneous electron transfer, favorable anti-fouling traits in the presence of a wide range of interferents, good stability, and reproducible analytical performance. Finally, the developed sensor was validated for real-time quantification of CRB from spiked water, food, and bio-samples, which depicted acceptable recoveries (98.6 to 101.5%) with RSD values between 0.35 and 2.23%. Further, to derive the possible sensing mechanism, the valence orbitals projected density of states (PDOS) for C, H, and N atoms of an isolated CRB molecule, VSe2 + CNT and VSe2 + CNT + CRB were calculated using density functional theory (DFT) calculations. The dominant charge transfer from the valence 2p-orbitals of the C and N atoms of CRB to CNT is responsible for the electrochemical sensing of CRB molecules.

Carbon quantum dots composite for enhanced selective detection of dopamine with organic electrochemical transistors.

A compact organic electrochemical transistors (OECT) sensor enriched with carbon quantum dots (CQDs) was developed to enhance the transconductance of an electropolymerized poly(3,4-ethylenedioxythiophene) (PEDOT) film, enabling the precise and selective detection of dopamine (DA). Accurate monitoring of DA levels is critical for diagnosing and managing related conditions. Incorporating CQDs, we have achieved a remarkable up to threefold increase in current at the DA detection peak in differential pulse voltammetry. This enhancement showcases superior selectivity even in the presence of high concentrations of interferents like uric acid and ascorbic acid. This material significantly boosts the sensitivity of OECTs for DA detection, delivering an amperometric response with a detection limit of 55 nM and a broader detection range (1 - 500 µM). Our results underscore the potential of low-dimensional carbonaceous materials in creating cost-effective, high-sensitivity devices for detecting DA and other biomolecules. This breakthrough sets the stage for the development of next-generation biosensors for point-of-care diagnostics.

A colorimetric and electrochemical dual-mode Hg2+ sensor utilizing oxidase-like activity arising from the combination of Hg2+ and palladium metal-organic framework@graphene.

Developing convenient and reliable methods for Hg2+ monitoring is highly important. Some precious metal nanomaterials with intriguing peroxidase-like activity have been used for highly sensitive Hg2+ detection. However, H2O2 must be added during these detections, which impedes practical applications of Hg2+ sensors due to its susceptible decomposition by environmental factors. Herein, we discovered that the combination of Hg2+ and palladium metal-organic framework@graphene (Pd-MOF@GNs) exhibits oxidase-like activity (OXD). In the absence of H2O2, this activity not only catalyzes the oxidation of chromogenic substrates such as 3,3',5,5'-tetramethylbenzidine (TMB) or o-phenylenediamine (OPD) to produce a color change but also enhances the electrical signals during OPD oxidation. Based on these properties, an effective and convenient dual-mode colorimetric and electrochemical sensor for Hg2+ has been developed. The colorimetric and amperometric linear relationships for Hg2+ were 0.045 µM-0.25 mM and 0.020 µM-2.0 mM, respectively. The proposed strategy shows good recovery in real sample tests, indicating promising prospects for multiple environmental sample detection of Hg2+ without relying on H2O2. The colorimetric and electrochemical dual-mode Hg2+ sensor is expected to hold great potentials in applications such as environmental monitoring, rapid field detection, and integration into smartphone detection of Hg2+.

In situ anchored ternary hierarchical hybrid nickel@cobaltous sulfide on poly(3,4-ethylenedioxythiophene)-reduced graphene oxide for highly efficient non-enzymatic glucose sensing.

A ternary hierarchical hybrid Ni@CoxSy/poly(3,4-ethylenedioxythiophene)-reduced graphene oxide (Ni@CoxSy/PEDOT-rGO) is rationally designed and in situ facilely synthesized as electrocatalyst to construct a binder-free sensing platform for non-enzymatic glucose monitoring through traditional electrodeposition procedure. The as-prepared Ni@CoxSy/PEDOT-rGO presents unique hierarchical structure and multiple valence states as well as strong and robust adhesion between Ni@CoxSy/PEDOT-rGO and GCE. Profiting from the aforementioned merits, the sensing platform constructed under optimal conditions achieved a wide detection range (0.2 µM ~ 2.0 mM) with high sensitivity (1546.32 µA cm-2 mM-1), a rapid response time (5 s), an ultralow detection limit (0.094 µM), superior anti-interference performance, excellent reproducibility and considerable stability. Furthermore, the sensor demonstrates an acceptable accuracy and appreciable recoveries ranging from 90.0 to 102.0% with less than 3.98% RSD in human blood serum samples, indicating the prospect of the sensor for the real samples analysis. It will provide a strategy to rationally design and fabricate ternary hierarchical hybrid as nanozyme for glucose assay.

Reusable graphite-based electrochemical sensors for L-dopa and dopamine detection.

A fully reusable electrochemical device is proposed for the first time made from laser cutting and a homemade conductive ink composed of carbon and nail polish. As a sensor substrate, we applied polymethyl methacrylate, which allows the surface to be renewed by simply removing and reapplying a new layer of ink. In addition to the ease of renewing the sensor's conductive surface, the design of the device has allowed for the integration of different forms of analysis. The determination of L-Dopa was performed using DPV, which presented a linear response range between 5.0 and 1000.0 µmol L-1, and a LOD of 0.11 µmol L-1. For dopamine, a flow injection analysis system was employed, and using the amperometric technique measurements were performed with a linear ranging from 2.0 to 100.0 µmol L-1 and a LOD of 0.26 µmol L-1. To demonstrate its applicability, the device was used in the quantification of analytes in pharmaceutical drug and synthetic urine samples.

Gel polymer electrolyte-based dual screen-printed electrodes for the headspace quantification of 4-ethylphenol and ethanethiol simultaneously in wines.

The identification and correction of negative factors, such as 4-ethylphenol and ethanethiol, is important to comply with food safety regulations and avoid economic losses to wineries. A simple amperometric measurement procedure that facilitates the simultaneous quantification of both compounds in the gas phase has been developed using fullerene and cobalt (II) phthalocyanine-modified dual screen-printed electrodes coated with a room temperature ionic liquid-based gel polymer electrolyte. The replacement of the typical aqueous supporting electrolyte by low-volatility ones improves both operational and storage lifetime. Under the optimum conditions of the experimental variables, Britton Robinson buffer pH 5 and applied potentials of + 0.86 V and + 0.40 V for each working electrode (vs. Ag ref. electrode), reproducibility values of 7.6% (n = 3) for 4-ethylphenol and 6.6% (n = 3) for ethanethiol were obtained, as well as capability of detection values of 23.8 µg/L and decision limits of 1.3 µg/L and 9.2 µg/L (α = ß = 0.05), respectively. These dual electrochemical devices have successfully been applied to the headspace detection of both compounds in white and red wines, showing their potential to be routinely used for rapid analysis control in wineries.

Multi-walled carbon nanotubes functionalized with a new Schiff base containing phenylboronic acid residues: application to the development of a bienzymatic glucose biosensor using a response surface methodology approach.

An innovative supramolecular architecture is reported for bienzymatic glucose biosensing based on the use of a nanohybrid made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with a Schiff base modified with two phenylboronic acid residues (SB-dBA) as platform for the site-specific immobilization of the glycoproteins glucose oxidase (GOx) and horseradish peroxidase (HRP). The analytical signal was obtained from amperometric experiments at - 0.050 V in the presence of 5.0 × 10-4 M hydroquinone as redox mediator. The concentration of GOx and HRP and the interaction time between the enzymes and the nanohybrid MWCNT-SB-dBA deposited at glassy carbon electrodes (GCEs) were optimized through a central composite design (CCD)/response surface methodology (RSM). The optimal concentrations of GOx and HRP were 3.0 mg mL-1 and 1.50 mg mL-1, respectively, while the optimum interaction time was 3.0 min. The bienzymatic biosensor presented a sensitivity of (24 ± 2) × 102 µA dL mg-1 ((44 ± 4) × 102 µA M-1), a linear range between 0.06 mg dL-1 and 21.6 mg dL-1 (3.1 µM-1.2 mM) (R2 = 0.9991), and detection and quantification limits of 0.02 mg dL-1 (1.0 µM) and 0.06 mg dL-1 (3.1 µM), respectively. The reproducibility for five sensors prepared with the same MWCNT-SB-dBA nanohybrid was 6.3%, while the reproducibility for sensors prepared with five different nanohybrids and five electrodes each was 7.9%. The GCE/MWCNT-SB-dBA/GOx-HRP was successfully used for the quantification of glucose in artificial human urine and commercial human serum samples.

Development of a Smart Wireless Multisensor Platform for an Optogenetic Brain Implant.

Implantable cell replacement therapies promise to completely restore the function of neural structures, possibly changing how we currently perceive the onset of neurodegenerative diseases. One of the major clinical hurdles for the routine implementation of stem cell therapies is poor cell retention and survival, demanding the need to better understand these mechanisms while providing precise and scalable approaches to monitor these cell-based therapies in both pre-clinical and clinical scenarios. This poses significant multidisciplinary challenges regarding planning, defining the methodology and requirements, prototyping and different stages of testing. Aiming toward an optogenetic neural stem cell implant controlled by a smart wireless electronic frontend, we show how an iterative development methodology coupled with a modular design philosophy can mitigate some of these challenges. In this study, we present a miniaturized, wireless-controlled, modular multisensor platform with fully interfaced electronics featuring three different modules: an impedance analyzer, a potentiostat and an optical stimulator. We show the application of the platform for electrical impedance spectroscopy-based cell monitoring, optical stimulation to induce dopamine release from optogenetically modified neurons and a potentiostat for cyclic voltammetry and amperometric detection of dopamine release. The multisensor platform is designed to be used as an opto-electric headstage for future in vivo animal experiments.