This study developed a dual-mode "on-off-on" sensor based on a bipyridine ruthenium metal-organic framework (Ru-MOF) and dual enzyme cleavage technology for the sensitive detection of the K-ras gene. The sensor combines electrogenerated chemiluminescence (ECL) and fluorescence (FL) detection modes, achieving high sensitivity and specificity in detecting the K-ras gene through catalytic hairpin assembly (CHA) and dual enzyme cleavage reactions. Experimental results showed that the detection limits for the K-ras gene were 0.044 fM (ECL) and 0.16 fM (FL), demonstrating excellent selectivity and stability during detection. Through testing actual samples, the sensor has shown potential for application in complex biological environments. This method offers an efficient and reliable new tool for cancer diagnosis and treatment.

The influence of incorporation of mitochondrial inner membrane potassium channel, and channel-forming peptide - Gramicidin on the ion transport and electromechanical properties of model lipid membranes tethered to gold electrode was electrochemically investigated by chronoamperometric and impedance spectroscopy techniques. In the case of the potassium channel the ion transport properties were modulated with channel-specific inhibitor - ATP-Mg complex, whereas in the case of gramicidin peptide - by replacing potassium with sodium ions. The observed two exponential current-time responses of the systems studied were interpreted in terms of ion penetration and electrostriction of tethered lipid bilayer membrane, and conclusions supported with the experiments on alkanethiol self-assembled monolayers of different alkanethiol chain lengths deposited on gold.

The KRAS G12C mutations, as crucial biomarkers, are closely associated with non-small cell lung cancer. Here, a novel label-free electrochemical biosensor with synergistic signal amplification of photocell energy transfer-reversible addition fragmentation chain transfer (PET-RAFT) and ring-opening polymerization (ROP) was developed for the first time for sensitive detection of KRAS G12C mutations. Specifically, hairpin DNA (hDNA), which act as biomolecular probe, was self-assembled on Au electrode surface by Au-S bond. 4-cyano-4-[(dodecylsulfanylthiocarbonyl) sulfanyl] pentanoic acid (CDTPA), the chain transfer agent of PET-RAFT reaction, was then attached to hDNA via amide bond. After that, the target DNA (tDNA) was captured on the electrode surface by complementary base pairing with hDNA. Subsequently, large numbers of electro-active monomers N-acryloxysuccinimide (NAS) were successfully grafted to the electrode surface via PET-RAFT reaction, which provided plenty of junction sites for doxorubicin-polycaprolactone (Dox-PCL) synthesized by ROP. Finally, the Dox-PCL was connected to the electrode surface by ester bond, significantly amplifying the electrochemical signal. Under optimized conditions, the biosensor has a wide linear detection range of 0.1 pM to 1 μM, with a detection limit of 86.9 fM. Attribute to its high sensitivity, specificity, reproducibility and stability, this biosensor possesses considerable potential in early diagnosis of disease and biomedical research.

A label-free electrochemical DNA detection strategy based on self-assembled α-FeO/FeO nanosheets with PNA-DNA hybridization process was developed for ultrasensitive detection of APOE ε4 gene, one of the most robust genetic risks for Alzheimer's Disease (AD). In this work, magnetic α-FeO/FeO heterogeneous nanosheets were prepared by hydrothermal-calcined reduction method and loaded with Au nanoparticles (AuNPs) on their surfaces. The magnetic α-FeO/FeO@Au nanocomposites significantly enhanced the electrochemical response as a signal amplification matrix and were able to bind to the magnetic glassy carbon electrode (MGCE) surface by magnetic self-assembly. Moreover, owing to the high specificity and stable binding capacity of PNA with respect to the target DNA, the biosensor not only enabled accurate (the limit of detection was estimated to be 0.147 pM) and rapid detection of the APOE ε4 gene, but also exhibited excellent specificity, stability and regeneration capability. Additional, the satisfactory recoveries were also obtained in real samples of human serum, ranging from 92.83 % to 106.22 % with relative standard deviation (RSD) between 0.25 % and 1.85 %. The results possessed important reference value for exploring the application of DNA biosensor technology in the diagnosis of APOE gene mutation.

Ti-6Al-4V (TC4) alloy is widely utilized as the structural material in marine industries owing to its low density, high specific strength, and favorable corrosion resistance. However, as biofouling drastically alters, some reported the major deleterious effect of bacteria has imposed a challenge to improve microbiologically influenced corrosion (MIC) resistance. A further opportunity for solving this problem is Cu micro-alloying, which was inspired by adding Cu for biomedical applications. Herein, a Ti-6Al-4V alloy with slight Cu addition (TC4-Cu) was exposed to 2216E media inoculated with Pseudomonas aeruginosa (P. A.), and then investigated compared to TC4. TC4-Cu exhibits lower corrosion current, more denser passive film, and lower weight loss with weaker pitting (a maximum pitting depth of 0.2 μm), compared to TC4 with a maximum pitting crater depth of 9.6 μm. Those demonstrated that the presence of Cu significantly improved the MIC resistance, and inhibited the proliferation of P. A., leading to a good antimicrobial efficacy against marine P. A. Moreover, besides the well-known bactericidal role, Cu ions were transferred to form CuO and CuO, constituting protective corrosion products, and thus improving the anti-microbial properties of TC4-Cu.

Cardiac troponin I (cTnI) has been widely used in clinical diagnosis of acute myocardial infarction (AMI). Herein, a sensitive electrochemical biosensor for cTnI analysis was designed, in which the simple synthesized Pd@PdPtCo mesoporous nanopolyhedras (MNPs) were utilized as signal amplifiers. The mesoporous polyhedral structure of Pd@PdPtCo MNPs endows them with more specific surface area and more active sites, as well as the synergistic effect between multiple metal elements, all of which increase the electrocatalytic performance of Pd@PdPtCo MNPs in efficiently oxidizing hydroquinone (HQ) to benzoquinone (BQ). Experimental results showed that Pd@PdPtCo MNPs had better performance in oxidation of HQ to BQ compared with their corresponding monometallic and bimetallic nanomaterials. With the aid of the interaction between antigens and antibodies, the peak current of HQ to BQ showed an upward trend with increasing concentration of cTnI, thus the quantitative detection of cTnI could be achieved. Under optimal conditions, the biosensor prepared in this work has a wider linear range (1.0 × 10-200 ng mL) and a lower detection limit (0.031 pg mL) than other sensors reported in literatures, coupled by good stability and high sensitivity. More importantly, it also performed well in complex serum environment, proving that the electrochemical sensor has a practical application potential in this field.

Multiple sclerosis (MS) is a recurrent inflammatory, demyelinating disease of the white matter in central nervous system (CNS). The number of MS patients is increasing, but the diagnostic process is still quite difficult, costly and requires combination of several methods. Myelin basic protein (MBP) makes up to 30 % of the myelin in CNS. It is known that MBP is released into the cerebrospinal fluid (CSF) as MS bioindicator. Herein, myelin specific DNA aptamer earlier developed for possible therapeutic purposes and anti-MBP antibody were applied as bioreceptors for MBP recognition on the same nanomodified sensor surfaces and their performances were compared. Biosensors were developed by using graphene oxide (GO) nanoparticles integrated onto pencil graphite electrodes (PGE) and bioreceptor molecules immobilized to create a bioactive layer for MBP binding. The measurements were run with electrochemical impedance spectroscopy (EIS). Selectivity of the biosensors was evaluated using human serum albumin (HSA). After optimization of binding parameters, biosensors were validated in artificial CSF. It was shown that LJM-5708 based aptasensor had LOD 0.65 ng/mL that was comparable to immunosensor LOD (0.36 ng/mL) in artificial CSF and showed its applicability in the clinical concentration range between 1 and 128 ng/mL.

Electrochemical impedance or admittance spectroscopy (EIS or EAS) has been widely used for decades, offering a label-free, rapid, real-time, and non-destructive assay for optically opaque and turbid bacterial solutions. However, PEF-induced changes in the bacterial envelope can present challenges in detecting the extent of membrane permeabilization in both Gram-positive and Gram-negative bacteria due to their distinct morphological properties. Here, we used a new approach for detecting bacterial membrane permeabilization induced by PEF using electrochemical admittance spectroscopy (EAS). The metabolic activity results have shown that the larger L. d. bulgaricus bacteria was found to be significantly more resistant to PEF strengths ranging from 4 to 16 kV/cm than the smaller E. coli bacteria at shorter PEF treatment durations (10 × 10 µs pulses). Interestingly, the difference in the increase of the admittance magnitude and a decrease in phase angle between the PEF treatment times of 10 × 10 µs and 10 × 100 µs pulses at different PEF strengths was more pronounced for E. coli bacteria samples. Our results demonstrate that EAS is more effective in comparing the degree of membrane permeabilization of Gram-positive and Gram-negative bacteria when longer PEF treatment durations are applied.

The core of bioelectrochemical systems (BESs) is electrochemically active microorganisms (EAMs), which exert spatial heterogeneity on electrode surface and influences BESs performance. Setting an optimal potential is an effective strategy for improving and optimizing BESs performance, however, how the electrode potential affects spatial structure of microbial community within anode biofilm is not known. Using a complex substrate-fed BES with a wastewater inoculum, this study investigated the community structure and composition of the stratified biofilm developed under the potential of -0.3 V, 0 V, +0.3 V and +0.6 V (vs. saturated calomel electrode) by freezing microtome method and high-throughput sequencing analysis. The spatial heterogeneity of biofilm community was found to be dependent on the electrode potential and a less stratified community structure was observed for +0.6 V than other potentials. Within the biofilms, the inner layers selected more Geobacter and the outer layers enriched more Acinetobacter and Serratia, potentially suggested a stratification of electron transfer pathway and metabolite-based interspecies communications. The results demonstrated the response of spatial heterogeneity of anode biofilm community to the change of electrode potential, which helps to understand the selectivity and enrichment of kinetically efficient anodic microbiome by electron potential.

In this study, we developed a new system that using zinc-based metal-organic frameworks NH-Zn-PTC as the donor and ZiF-8@PDA as the acceptor to achieve highly sensitive detection of carcinoembryonic antigen (CEA), using the fundamentals of electrochemiluminescence resonance energy transfer (ECL-RET). Firstly, the aggregation-induced quenching effect (ACQ) was eliminated by the coordination of PTC in MOF and the ECL signal was improved. Secondly, the ECL signal was further amplified by using Au NPs and amino groups as co-reaction promoters to generate more SO. In addition, the introduction of ZiF-8@PDA as an acceptor and NH-Zn-PTC as a donor took advantage of the feature of partial overlap of the UV-vis absorption spectrum and ECL emission spectra between the two, thereby effectively initiating the ECL-RET behavior, which improved the detection sensitivity of the sensor. The prepared immunosensor showed good linearity in the concentration range of 10 to 80 ng/mL with a detection limit of 18.20 fg/mL. This makes it promising for clinical testing of tumor markers.

In this work, the dual-mode aptasensor based on 3D porous AuNPs/MXene using "turn-on" electrochemical method and "turn-off" fluorescent strategy was fabricated. Here, 2D MXene was processed into 3D porous MXene by sacrificial polymethylmethacrylate (PMMA) spherical template. And the meteor hammer-like AuNPs which had good electrochemical properties and quenching effect on fluorescence was synthesized by single electrodeposition. Dual-signal labeled Nile Blue (NB) was in situ grafted to the Hg aptamer ends of 3D porous AuNPs/MXene/GCE, and an efficient and sensitive signal interface was constructed to realize the sensitive detection of Hg. 3D porous AuNPs/MXene had the advantages of large specific surface area, excellent electron transmission performance and signal amplification. The experimental results indicated that this sensor exhibited high sensitivity to Hg in both electrochemical and fluorescent sensing, with detection limits of 2.69 fM and 1.60 fM, respectively. Further, the dual-mode aptasensor can ensure the detection accuracy and target quantization. The dual-mode aptasensor has been successfully applied to the ultra-trace detection of Hg in actual water samples, which shows the potential of aptamer sensor in detecting heavy metal ions in environmental monitoring.

C-reactive protein (CRP) is a critical biomarker for detecting inflammation and forecasting cardiovascular disease. We present an advanced electrochemical immunosensor utilizing laser-induced graphene (LIG)/MXene-gold nanoparticles (Mx-AuNPs) electrode for CRP detection. The Mx-AuNPs nanocomposite, synthesized via in-situ reduction of HAuCl by MXene, leverages MXene's reducing properties for effective nanoparticle deposition, confirmed through scanning electron microscopy. This electrode demonstrates superior electrochemical performance due to enhanced surface area and synergy between LIG and Mx-AuNPs, improving overall electrode conductivity. The A-CRP antibody, immobilized via a cysteamine linker, enables CRP detection. The immunosensor achieves excellent detection across 10 pg mL to 10 µg mL CRP, with a low detection limit of 1.45 pg mL, and shows high selectivity for CRP. This LIG/Mx-AuNPs-based immunosensor is promising for sensitive CRP detection, aiding early cardiovascular disease diagnosis and improving patient outcomes.

The impact of four clinically significant genetic variants of endothelial nitric oxide synthase (eNOS) polymorphisms on the concentrations of nitric oxide [NO] and peroxynitrite [ONOO] has been given scant consideration. This study utilized a [NO]/[ONOO] ratio to determine the extent of endothelial dysfunction caused by these variations in the eNOS gene. The single nucleotide polymorphisms (T-786C, C-665T, and Glu298Asp) and a variable number of tandem repeats (intron 4 a/b/c) were genotyped in human umbilical vein endothelial cells (HUVEC), using sanger sequencing and DNA electrophoresis, respectively. Nanosensors were used to determine the maximal [NO] and [ONOO], while traditional and low-temperature SDS-PAGE were used to evaluate the expression of eNOS and the eNOS dimer-to-monomer ratio, respectively. The study results indicate that the eNOS haplotype H3 (G T/C C 4a/c allele) may have a protective effect against cardiovascular disease (CVD) with the [NO]/[ONOO] ratio higher than 2. However, the eNOS haplotypes H2 (G T/C C 4a/b) and H5 (T T/C C 4b) increase the susceptibility to CVD with [NO]/[ONOO] ratio lower than 1. The results suggest that certain eNOS genetic variants may influence susceptibility to cardiovascular disease (CVD) while other variants may have a protective effect.

In modern society, due to the sharp increase in pollutants that cause DNA damage, there is a growing demand for innovative detection techniques and biomarkers. In this paper, the electrochemical behavior of HepG2 cells exposed to CdCl was investigated, and the electrochemical response mechanism of DNA damage was identified by exploring the correlation between the DNA damage response and purine metabolism. Western blot analysis revealed that the expression levels of ATM and Ku70 increased at 0.3 μM CdCl, indicating a DNA damage response and activation of DNA repair processes. Simultaneously, elevated expression levels of PRPP aminotransferase, HPRT, and XOD were observed, leading to an increase in intracellular purine levels and electrochemical signals. The expression of Ku70 peaked at 0.5 μM CdCl, indicating the highest DNA repair activity. The expression profiles of these purine metabolism proteins mirrored those of Ku70, suggesting a strong correlation between the activation of purine metabolism and DNA damage repair. Consistently, intracellular purine levels exhibited a similar trend, leading to corresponding changes in electrochemical signals. In summary, electrochemical using intracellular purines as biomarkers has the potential to emerge as a novel method for detecting early DNA damage.

Monitoring of the levels of 5-hydroxyindole-3-acetic acid (5-HIAA) is of significant importance for diagnostics of carcinoid tumors. We propose simple catalytic electrochemical sensors for the determination of 5-HIAA in urine using laccase and its mimetics. Laccase-like nanozymes (LacNZs) were synthesized via a chemical reduction, and resulting PtMn and MnO nanoflowers (NFs) demonstrated laccase-like activity similar to the laccase from the Trametes zonata. In addition, these LacNZs showed enhanced stability under a wide range of pH (3.0-7.5), temperatures (4-70 °C), and ionic strengths (up to 500 mM NaCl). The developed PtMn NF/graphite electrode, similar to a laccase/graphite electrode, can detect 5-HIAA with a high sensitivity (25 000 ± 12 A·M·m and 1900 ± 9 A·M·m, respectively) and have linear ranges of 0.3 - 15 μM and 2 - 50 μM. The sensors work at low working potentials with a detection limit of 0.16 and 1.4 μM, covering the normal and pathologic ranges of 5-HIAA (1 - 50 μM) content in urine. They have been successfully applied to 5-HIAA assay in urine samples of people with various diseases and revealed good recovery values and reproducibility. Additionally, the LacNZ-sensor has the best stability and can be used up to 20 days.

CRISPR/Cas9-mediated gene editing offers promising and safe therapeutic options for a wide range of diseases. The technical difficulty of efficiently acquiring large quantities of gene-edited therapeutic cells in a short time period is now preventing the widespread clinical application of CRISPR/Cas9-mediated gene editing. Herein, a Large Volume Continuous Electroporation Chip (LaViE-Chip) has been developed to address the challenge of acquiring sufficient quantities of genetically edited cells for CRISPR/Cas9 gene editing. By connecting multiple relatively narrow microfluidic channels in parallel, a satisfactory balance between cell flow volume and electric field uniformity was achieved with two simple off-chip electrodes, which also isolated harmful effects around electrodes from target cells. Meanwhile, by carefully designing the curvature of the microfluidic channel, hydrodynamic controlled rotation of target cells has been realized to improve the transfection efficiency and cell viability. With these improvements, the LaViE-Chip realized 71.06 % electrotransfection efficiency, 84.3 % cell viability, and 10 cell/min cell processing speed. Moreover, the first successful incessant CRISPR gene editing by electroporation has been demonstrated, laying the technical foundation of therapeutic CRISPR gene editing.

Edible biosensors can measure a wide range of physiological and biochemical parameters, including temperature, pH, gases, gastrointestinal biomarkers, enzymes, hormones, glucose, and drug levels, providing real-time data. Edible biocatalytic biosensors represent a new frontier within healthcare technology available for remote medical diagnosis. The main challenges to develop edible biosensors are: i) finding edible materials (i.e. redox mediators, conductive materials, binders and biorecognition elements such as enzymes) complying with Food and Drug Administration (FDA), European Food Safety Authority (EFSA) and European Medicines Agency (EMEA) regulations; ii) developing bioelectronics able to operate in extreme working conditions such as low pH (∼pH 1.5 gastric fluids etc.), body temperature (between 37 °C and 40 °C) and highly viscous bodily fluids that may cause surface biofouling issues. Nowadays, advanced printing techniques can revolutionize the design and manufacturing of edible biocatalytic biosensors. This review outlines recent research on biomaterials suitable for creating edible biocatalytic biosensors, focusing on their electrochemical properties such as electrical conductivity and redox potential. It also examines biomaterials as substrates for printing and discusses various printing methods, highlighting challenges and perspectives for edible biocatalytic biosensors.

We present the novel design of photosystem I (PSI)-based biosolar cell, whereby conductive transparent electrode materials, such as ITO or FTO, are replaced with glass covered with silver island film. This nanostructured metallic layer combines high electric conductance with enhancing the absorption efficiency of the PSI biocatalyst via the plasmonic effect. We demonstrate strong enhancement of the photocurrent generated in the biohybrid electrode composed of oriented layers of PSI reaction centers due to plasmonic interactions of the PSI fluorophores and redox centres with the conductive silver island film.

Microbiologically influenced corrosion (MIC) in shale gas field is a major threat with the hydraulic fracturing fluid injected into the subsurface. In this study, the microbiome collected from a shale gas produced water sample was extracted and cultivated in ATCC 1249 medium modified with 10 g/L NaCl anaerobically at 30 °C. d-amino acids, which were reported as biocide enhancers, were found to enhance 2,2-dibromo-3-nitrilopropionamide (DBNPA) biocide on the mitigation of shale microbiome MIC on X80 carbon steel. The combination of 50 ppm (w/w) d-leucine + 50 ppm d-alanine + 1 ppm d-tyrosine had the best enhancement effect on 50 ppm DBNPA with 84 % less weight loss, and 67 % lower corrosion current density (i) compared to 50 ppm DBNPA alone. The corrosion data were consistent with the enhanced biofilm inhibition observation. The experimental data also indicated that d-tyrosine used alone at a low dosage of 1 ppm enhanced DBNPA considerably, with 44 % less weight loss and 47 % less i. The electrochemical results showed the positive response of shale gas microbiome biofilm to the injected magnetite nanoparticles indicating the extracellular electron transfer might be a main mechanism for its corrosion.