A Fully Integrated Handheld Electrochemical Sensing Platform for Point-of-Care Testing of Escherichia coli O157:H7.

Fully integrated devices that enable full functioning execution without or with minimum external accessories or equipment are deemed to be one of the most desirable and ultimate objectives for modern device design and construction. Escherichia coli O157:H7 (E. coli O157:H7) is often linked to outbreaks caused by contaminated water and food. However, the sensors that are currently used for point-of-care E. coli O157:H7 (E. coli O157:H7) detection are often large and cumbersome. Herein, we demonstrate the first example of a handheld and pump-free fully integrated electrochemical sensing platform with the capability to point-of-care test E. coli O157:H7 in the actual samples of E. coli O157:H7-spiked tap water and E. coli O157:H7-spiked watermelon juice. This platform was made possible by overcoming major engineering challenges in the seamless integration of a microfluidic module for pump-free liquid sample collection and transportation, a sensing module for efficient E. coli O157:H7 testing, and an electronic module for automatically converting and wirelessly transmitting signals into a single and compact electrochemical sensing platform that retains its inimitable stand-alone, handheld, pump-free, and cost-effective feature. Although our primary emphasis in this study is on detecting E. coli O157:H7, this pump-free fully integrated handheld electrochemical sensing platform may also be used to monitor other pathogens in food and water by including specific antipathogen antibodies.

Universal Fully Integrated Wearable Sensor Arrays for the Multiple Electrolyte and Metabolite Monitoring in Raw Sweat, Saliva, or Urine.

Fully integrated wearable sensors are capable of dynamically, directly, and independently tracking biomarkers in raw noninvasive biofluids without any other equipment or accessories by integrating the unique on-body monitoring feature with the special complete functional implementation attribute. Sweat, saliva, and urine are three important noninvasive biofluids, and changes in their biomarkers hold great potential for revealing physiological conditions. However, it is still a challenge to design single fully integrated wearable sensor arrays (FIWSAs) that are universally able to concurrently measure electrolytes and metabolites in three of the most common noninvasive biofluids including sweat, saliva, and urine. Here, we propose the first single universal FIWSAs for wirelessly, noninvasively, and simultaneously measuring various metabolites (i.e., uric acid) and electrolytes (i.e., Na+ and H+) in raw sweat, saliva, or urine under subjects' exercise by integrating the specifically designed microfluidic, sensing, and electronic modules in a seamless manner. We evaluate its utility for noninvasive gout management in healthy subjects and in gout patients through a purine-rich meal challenge and with a medicine-treatment control, respectively. Noninvasive monitoring of multiple electrolytes and metabolites in a variety of raw noninvasive biofluids via such single universal FIWSAs may enrich the understanding of the biomarkers' levels in the body and would also facilitate self-health management.

Microporous P-doped carbon spheres sensory electrode for voltammetry and amperometry adrenaline screening in human fluids.

An electrochemical sensor-based phosphorus-doped microporous carbon spheroidal structures (P-MCSs) has been designed for selective adrenaline (ADR) signaling in human blood serum. The P-MCS electrode sensor is built with heterogeneous surface alignments including multiple porous sizes with open holes and meso-/macro-grooves, rough surface curvatures, and integral morphology with interconnected and conjugated microspheres. In addition, the P atom-doped graphitic carbon forms highly active centers, increases charge mobility on the electrode surface, creates abundant active centers with facile functionalization, and induces binding to ADR molecules. The designed P-MCS electrode exhibits ultrasensitive monitoring of ADR with a low detection limit of 0.002 µM and high sensitivity of 4330 µA µM-1 cm-2. In addition, two electrochemical techniques, namely, square wave voltammetry (SWV) and chronoamperometry (CA), were used; these techniques achieve high stability, fast response, and a wide linear range from 0.01 to 6 µM. The sensing assays based on P-MCSs provide evidence of the formation of active interfacial surface-to-ADR binding sites, high electron diffusion, and heavy target loads along with/without a plane of spheroids. Thus, P-MCSs can be used for the routine monitoring of ADR in human blood serum, providing a fast response, and requiring highly economical materials at extremely low concentrations. Electrode surface modulation based on P-doped carbon spheres (P-MCS) exhibits high electrochemical activity with fast charge transport, multi-diffusible active centers, high loading of ADR, and facile molecular/electron diffusion at its surface. The P-MCS sensitively and selectively detects the ADR in human fluids and can be used for clinical investigation of some neuronal diseases such as Alzheimer diseases.

Subchronic Microcystin-LR Aggravates Colorectal Inflammatory Response and Barrier Disruption via Raf/ERK Signaling Pathway in Obese Mice.

Microcystin-LR (MC-LR) is an extremely poisonous cyanotoxin that poses a threat to ecosystems and human health. MC-LR has been reported as an enterotoxin. The objective of this study was to determine the effect and the mechanism of subchronic MC-LR toxicity on preexisting diet-induced colorectal damage. C57BL/6J mice were given either a regular diet or a high-fat diet (HFD) for 8 weeks. After 8 weeks of feeding, animals were supplied with vehicle or 120 µg/L MC-LR via drinking water for another 8 weeks, and their colorectal were stained with H&E to detect microstructural alterations. Compared with the CT group, the HFD and MC-LR + HFD-treatment group induced a significant weight gain in the mice. Histopathological findings showed that the HFD- and MC-LR + HFD-treatment groups caused epithelial barrier disruption and infiltration of inflammatory cells. The HFD- and MC-LR + HFD-treatment groups raised the levels of inflammation mediator factors and decreased the expression of tight junction-related factors compared to the CT group. The expression levels of p-Raf/Raf and p-ERK/ERK in the HFD- and MC-LR + HFD-treatment groups were significantly increased compared with the CT group. Additionally, treated with MC-LR + HFD, the colorectal injury was further aggravated compared with the HFD-treatment group. These findings suggest that by stimulating the Raf/ERK signaling pathway, MC-LR may cause colorectal inflammation and barrier disruption. This study suggests that MC-LR treatment may exacerbate the colorectal toxicity caused by an HFD. These findings offer unique insights into the consequences and harmful mechanisms of MC-LR and provide strategies for preventing and treating intestinal disorders.

Artificial enzyme innovations in electrochemical devices: advancing wearable and portable sensing technologies.

With the rapid evolution of sensing technologies, the integration of nanoscale catalysts, particularly those mimicking enzymatic functions, into electrochemical devices has surfaced as a pivotal advancement. These catalysts, dubbed artificial enzymes, embody a blend of heightened sensitivity, selectivity, and durability, laying the groundwork for innovative applications in real-time health monitoring and environmental detection. This minireview penetrates into the fundamental principles of electrochemical sensing, elucidating the unique attributes that establish artificial enzymes as foundational elements in this field. We spotlight a range of innovations where these catalysts have been proficiently incorporated into wearable and portable platforms. Navigating the pathway of amalgamating these nanoscale wonders into consumer-appealing devices presents a multitude of challenges; nevertheless, the progress made thus far signals a promising trajectory. As the intersection of materials science, biochemistry, and electronics progressively intensifies, a flourishing future seems imminent for artificial enzyme-infused electrochemical devices, with the potential to redefine the landscapes of wearable health diagnostics and portable sensing solutions.

Enzymeless copper microspheres@carbon sensor design for sensitive and selective acetylcholine screening in human serum.

Follow up of neuronal disorders, such as Alzheimer's and Parkinson's diseases using simple, sensitive, and selective assays is urgently needed in clinical and research investigation. Here, we designed a sensitive and selective enzymeless electrochemical acetylcholine sensor that can be used in human fluid samples. The designed electrode consisted of a micro spherical construction of Cu-metal decorated by a thin layer of carbon (CuMS@C). A simple and one-pot synthesis approach was used for Cu-metal controller formation with a spherical like structures. The spherical like structure was formed with rough outer surface texture, circular build up, homogeneous formation, micrometric spheres size (0.5 -1 µm), and internal hollow structure. The formation of a thin layer of carbon materials on the surface of CuMS sustained the catalytic activity of Cu atoms and enriched negatively charge of the surface. CuMS@C acted as a highly active mediator surface that consisted of Cu metal as a highly active catalyst and carbons as protecting, charge transport, and attractive surface. Therefore, the CuMS@C surface morphology and composition played a key role in various aspects such as facilitated ACh diffusion/loading, increased the interface surface area, and enhanced the catalytic activity. The CuMS@C acted as an electroactive catalyst for ACh electrooxidation and current production, due to the losing of two electrons. The fabricated CuMS@C could be a highly sensitive and selective enzymeless sensor for detecting ACh with a detection limit of 0.1 µM and a wide linear range of 0.01 - 0.8 mM. The designed ACh sensor assay based on CuMS@C exhibited fast sensing property as well as sensitivity, selectivity, stability, and reusability for detecting ACh in human serum samples. This work presents the design of a highly active electrode surface for direct detection of ACh and further clinical investigation of ACh levels.

Nitrogen-doped carbon hollow trunk-like structure as a portable electrochemical sensor for noradrenaline detection in neuronal cells.

To date, the production and development of portable analytical devices for environmental and healthcare applications are rapidly growing. Herein, a portable electrochemical sensor for monitoring of noradrenaline (NA) secreted from living cells using mesoporous carbon-based materials was fabricated. The modification of the interdigitated electrode array (IDA) by nitrogen-doped mesoporous carbon spheres (N-doped MCS) and nitrogen-doped carbon hollow trunk-like structure (N-doped CHT) was used to fabricate the NA sensor. The N-doped CHT surface shows multiple holes distributed with micrometer-sized open holes (1-2 µm) and nanometer-sized thin walls (∼98 nm). The N-doped CHT surface heterogeneity of wrinkled and twisted hollow trunk structures improve the diffusion pathway and the NA molecules loading. The N-doped CHT/IDA showed a highly selective assay for monitoring of NA with high sensitivity (1770 µA/µM × cm2), a low detection limit (0.005 µM), and a wide linear range (0.01-0.3 µM). The N-doped CHT/IDA monitored the NA secreted from PC12 cells under various concentrations of simulation agents (KCl). The designed N-doped CHT/IDA provides a portable NA-sensor assay with facile signaling, good stability, high biocompatibility, in-vitro assay compatibility, and good reproducibility. Therefore, the designed sensor can be used as a portable sensor for NA detection in live cells and can be matched with portable smartphones after further developments.

Non-metal sensory electrode design and protocol of DNA-nucleobases in living cells exposed to oxidative stresses.

Sensory protocols for evaluation of DNA distortion due to exposure to various harmful chemicals and environments in living cells are needed for research and clinical investigations. Here, a design of non-metal sensory (NMS) electrode was built by using boron-doped carbon spherules for detection of DNA nucleobases, namely, guanine (Gu), adenine (Ad), and thymine (Th) in living cells. The key-electrode based nanoscale NMS structures lead to voids with a facile diffusion, and strong binding events of the DNA nucleobases. Furthermore, the NMS geometric structures would significantly create electrode surfaces with numerous centrally active sites, curvature topographies, and anisotropic spherules. The NMS shows potential as sensitive protocol for DNA-nucleobases in living cells exposed to oxidative stresses. In one-step signaling assay, NMS shows high signaling transduction of Gu-, Ad-, and Th-DNA nucleobases targets with ultra-sensitive and low detection limits of 3.0, 0.36, and 0.34 nM, respectively, and a wide linear range of up to 1 µM. The NMS design and protocol show evidence of the role of surface construction features and B-atoms incorporated into the graphitic carbon network for creating abundant active sites with facile electron diffusion and heavily target loads along with within-/out-plane circular spheres. Indeed NMS, with spherule-rich interstitial surfaces can be used for sensitive and selective evaluation of damaged-DNA to various dysfunctional metabolism in the human body.

Three-Dimensional Circular Surface Curvature of a Spherule-Based Electrode for Selective Signaling and Dynamic Mobility of Norepinephrine in Living Cells.

A highly sensitive protocol for signaling norepinephrine (NEP) in human fluids and neuronal cell line models should be established for clinical investigation of some neuronal diseases. A metal-free electrode catalyst was designed based on a sulfur-doped carbon spheroidal surface (S-CSN) and employed as a transducing element for selective signaling of NEP in biological samples. The designed electrode of S-CSN features a spherical construct and curvature surface to form a spheroidal nanolayer with an average layer size of <2 nm. S-CSN shows surface topography of a circular surface curvature with a rugged surface texture, ridge ends, and free open spaces between interlayers. The rich-space diversity surfaces offer highly active surface with facile molecular/electron diffusion, multi-diffusive centers, and high target loading along with in-/out-of-plane circular spheres of the S-CSN surface. The active doping of S atoms onto the carbon-based electrode creates an active transducing element with many active sites, strong binding to targeted molecules, facile diffusion of charges/molecules, long-term durability, and dense reactive exposure sites for signaling NEP at ultratrace levels. S-CSN could be a sensitive and selective nanosensor for signaling NEP and establishing a sensing protocol with high stability and reproducibility. The sensory protocol based on S-CSN exhibits high sensitivity and selectivity with a low detection limit of 0.001 µM and a wide linear range of 0.01-0.8 µM. The in vitro sensory protocol for NEP secreted from living cells (neuronal cell line model) under stimulated agents possesses high sensitivity, low cytotoxicity, and high biocompatibility. These results confirm the successful establishment of NEP sensor in human blood samples and neuronal cells for clinical investigation.

One-step selective screening of bioactive molecules in living cells using sulfur-doped microporous carbon.

A metal-free electrode using heteroatom-doped microporous carbon was fabricated for the ultrasensitive monitoring of mono-bioactive molecules and the selective signaling of dopamine (DA) secreted by living cells. The constructed electrode based on sulfur-doped microporous carbon (S-MC) shows a high surface area, a spherical construction, numerous carbon chain defects, and microporous structures, which are the key factors of the interactive signaling transducer, fast response, and active interfacial surfaces. The intrinsic features of S-MC with different %S-doping (S-MC-1, and S-MC-2) through the sp2-carbon chain create abundant catalytic active sites, facilitate molecular diffusion through the microporous structure, promote strong binding with the targeted molecules, and induce interactions at electrolyte-electrode interfaces. The S-MC-1 provides selective signaling in a tertiary mixture of DA, ascorbic acid (AA), and uric acid (UA) with a high sensitivity and a wide linear range of 0.01-5, 10-4000, and 1-2000 µM, respectively. The detection limits were set at 3 nM, 1.26 µM, and 0.23 µM for DA, AA, and UA respectively. The S-MC-1 demonstrated a selective screening of DA released from PC12 cells under a K+ ion- stimulator with high sensitivity and promoted high biocompatibility, low cytotoxicity, high stability, and reliable reproducibility (%RSD ranged from 1 to 2.7). Our findings indicated that the S-MC-1 can be utilized as an in-vitro model for simultaneously monitoring extracellular-DA secreted from living cells and sensing mono-bioactive molecules in biological samples.

3D-Ridge Stocked Layers of Nitrogen-Doped Mesoporous Carbon Nanosheets for Ultrasensitive Monitoring of Dopamine Released from PC12 Cells under K+ Stimulation.

3D-ridge nanosheets of N-doped mesoporous carbon (NMCS)-based electrodes are fabricated as ultrasensitive biosensors for in vitro monitoring of dopamine (DA) released from living cells. The large-scale ranges of dense-layered sheets are arranged linearly with a thickness of <10 nm, soft tangled edges, stocked layer arrangements, and tunable mesoporous frameworks with 3D orientations. The intrinsic features of the active interfacial surface of the electrode based on NMCS along with polarized surfaces, dense surface-charged matrices, fast electron transfer, and easy molecular diffusion, are present in the highly active electrode for biosensing applications. The designed electrode based on the NMCS shows high sensitivity and selectivity for DA sensing even in the presence of physiological interference molecules, such as ascorbic acid and/or uric acid, at a low applied potential of 0.25 V versus Ag/AgCl. The large-scale NMCS-based electrode shows low detection limits as low as 10 nmol L-1 , wide linear range up to 0.5 mmol L-1 , long-term stability for more than 15 d (relative standard deviation (RSD)= 5.8%), and a low cytotoxicity with high biocompatibility. The findings demonstrated that the NMCS-based electrode is a reliable modified electrode for ultratrace sensitivity of DA, which is secreted normally from dopaminergic cells (PC12) or under a stimulating agent (K+ ).

Broccoli-shaped biosensor hierarchy for electrochemical screening of noradrenaline in living cells.

Monitoring and determination of ultra-trace concentrations of monoamine neurotransmitter such as noradrenaline (NA) in living cells with simple, sensitive and selective assays are significantly interesting. We design NA-electrode sensing system based on C-, N-doped NiO broccoli-like hierarchy (CNNB). The spherical broccoli-head umbrella architectures associated with nano-tubular arrangements enabled to tailor NA biosensor design. The homogenous doping and anisotropic dispersion of CN nanospheres along the entire NB head nanotubes lead to creating of abundant electroactive sites in the interior tubular vessels and outer surfaces for ultrasensitive detection of NA in living cells such as PC12. The CNNB biosensor electrodes showed efficient electrocatalytic activity, enhanced kinetics for electrooxidation of NA, and fast electron-transfer between electrode-electrolyte interface surfaces, enabling synergistic enhancement in sensitivity, and selectivity at a low-detectable concentration of ∼ 6nM and reproducibility of broccoli-shaped NA-electrodes. The integrated CNNB biosensor electrodes showed evidence of monitoring and screening of NA released from PC12 cells under K+ ion-extracellular stimulation process. The unique features of CNNB in terms of NA-selectivity among multi-competitive components, long-term stability during the detection of NA may open their practical, in-vitro application for extracellular monoamine neurotransmitters detection in living cells.

Fabrication of photo-electrochemical biosensors for ultrasensitive screening of mono-bioactive molecules: the effect of geometrical structures and crystal surfaces.

The controlled design of biosensors based on the photo-electrochemical technique with high selectivity, sensitivity, and rapid response for monitoring of mono-bioactive molecules, particularly dopamine (DA) levels in neuronal cells is highly necessary for clinical diagnosis. Hierarchical carbon-, nitrogen-doped (CN) nickel oxide spear thistle (ST) flowers associated in single-heads (S), and symmetric and asymmetric-double heads (D and A, respectively) that are tightly connected through a micrometric dipole-like rod or trunk were fabricated by using a simple synthetic protocol. The CN-ST flower heads were decorated with dense nano-tubular like hedgehog needle skins in vertical alignments. These designated architectures are key features for creating biosensor surface electrodes for photo-electrochemical, ultrasensitive screening of mono-bioactive molecules. The exceptional electrode designs produced numerous catalytically active sites, large surface area, and high electron-transfer mobility. The active coating of carbon-nitrogen nanospheres significantly enhanced the photo-electrocatalytic activity of the prepared biosensor electrodes and prevented leakage of photocatalytic activity under long-term exposure to irradiation. Among all photo-electrochemical assays, the biosensors showed significant sensitivity and selectivity for DA in the presence of interfering molecules such as ascorbic acid (AA), uric acid (UA), adrenaline (A), and noradrenaline (NA). The photo-electrochemical property of the CN-SST-{110} crystal surface electrode showed significant sensing performance for DA in terms of unimpeded diffusion pathways, a wide concentration-detection range, and a low detection limit, even in the presence of potentially interfering molecules compared with other electrode-modified CN-DST-{111} and CN-AST-{101} crystal surfaces. Furthermore, the CN-SST photo-biosensor electrode shows potential in the selective and sensitive determination of DA in real samples, such as human serum and secreted DA from living cells. This finding indicates that the hierarchical ST biosensor may enable analytical discrimination and monitoring of DA and can be employed for clinical diagnosis application.