Complete removal of heavy metals with simultaneous efficient treatment of etching terminal wastewater using scaled-up microbial electrolysis cells

The treatment of actual low and high strengths etching terminal wastewater (ETW) from plating and electronic industry meeting national discharge standards is demonstrated in laboratory scale (1 L) and in scaled-up (40 L) microbial electrolysis cells (MECs). Both cylindrical single-chamber MECs achieved complete removal of heavy metals and efficient treatment of organics using either low strength ETW at an hydraulic retention time (HRT) of 5 d, or high strength wastewater at HRTs of 7 d (1 L) or 9 d (40 L). The removal rate of organics and heavy metals increased by 36-fold and scaled almost with the reactor volume ratio of 40. Electrode potentials in the scaled-up MECs (40 L) were resilient to the wastewater strength. Bacterial communities on both anodes and cathodes of the 1 L and the 40 L reactors experienced a selective shock and a significant community change after switching from low to high strengths wastewater, although reactor performance was effectively maintained. This study demonstrates complete removal of multiple heavy metals with simultaneous efficient wastewater treatment in MECs of different scales meeting China national discharge standards and provides a plausible approach for simultaneous removal of value-added products (e.g., heavy metals) and efficient treatment of practical etching terminal wastewater

Efficient conversion of bicarbonate (HCO3⁻) to acetate and simultaneous heavy metal Cr(VI) removal in photo-assisted microbial electrosynthesis systems combining WO3/MoO3/g-C3N4 heterojunctions and Serratia marcescens electrotroph

The removal of the hazardous heavy metal Cr(VI) in water and the simultaneous production of acetate from the reduction of inorganic carbon (HCO3−) is demonstrated in a photo-assisted microbial electrosynthesis (MES) system incorporating WO3/MoO3/g-C3N4 Z-scheme heterojunctions and Serratia marcescens Q1 electrotroph cathode. The rates of acetate production (6.1 ± 0.3 mg/L/h) and Cr(VI) removal (4.5 ± 0.1 mg/L/h) recorded at a circuital current of 2.8 ± 0.1 A/m2 were 2.4-fold (acetate production), 1.7-time (Cr(VI) removal) and 1.6-fold (circuital current) of those in the controls recorded in the absence of WO3/MoO3/g-C3N4, and 1.6-fold (acetate production) and 1.8-time (circuital current) of those in the absence of both Cr(VI) and WO3/MoO3/g-C3N4. Photogenerated WO3/MoO3/g-C3N4 conduction bands electrons favored both direct or indirect (via S. marcescens) reductions of Cr(VI) and H+, with the latter producing H2 which was further metabolized by S. marcescens with HCO3− to yield acetate. The higher circuital current drawn under photoirradiation conditions refilled the photo-generated valence band holes in the semiconductor and provided the driving force for the reduction reactions. This study provides an alternative and feasible approach for achieving complete removal of toxic heavy metal from water and industrial waters with simultaneous conversion of inorganic carbon to key block chemicals

High-Sensitivity and High-Efficiency Detection of DNA Hydroxymethylation in Genomic DNA by Multiplexing Electrochemical Biosensing

DNA hydroxymethylation (5-hmC) is a kind of new epigenetic modification, which plays key roles in DNA demethylation, genomic reprogramming, and the gene expression in mammals. For further exploring the functions of 5-hmC, it is necessary to develop sensitive and selective methods for detecting 5-hmC. Herein, we developed a novel multiplexing electrochemical (MEC) biosensor for 5-hmC detection based on the glycosylation modification of 5-hmC and enzymatic signal amplification. The 5-hmC was first glycosylated by T4 β-glucosyltransferase and then oxidated by sodium periodate. The resulting glucosyl-modified 5-hmC (5-ghmC) was incubated with ARP-biotin and was bound to avidin-HRP. The 5-hmC can be detected at the subnanogram level. Finally, we performed 5-hmC detection for mouse tissue samples and cancer cell lines. The limit of detection of the MEC biosensor is 20 times lower than that of commercial kits based on optical meaurement. Also, the biosensor presented high detection specificity because the chemical reaction for 5-hmC modification can not happen at any other unhydroxymethylated nucleic acid bases. Importantly, benefited by its multiplexing capacity, the developed MEC biosensor showed excellent high efficiency, which was time-saving and cost less

DNA Nanostructure-Based Universal Microarray Platform for High-Efficiency Multiplex Bioanalysis in Biofluids

Microarrays of biomolecules have greatly promoted the development of the fields of genomics, proteomics, and clinical assays because of their remarkably parallel and high-throughput assay capability. Immobilization strategies for biomolecules on a solid support surface play a crucial role in the fabrication of high-performance biological microarrays. In this study, rationally designed DNA tetrahedra carrying three amino groups and one single-stranded DNA extension were synthesized by the self-assembly of four oligonucleotides, followed by high-performance liquid chromatography purification. We fabricated DNA tetrahedron-based microarrays by covalently coupling the DNA tetrahedron onto glass substrates. After their biorecognition capability was evaluated, DNA tetrahedron microarrays were utilized for the analysis of different types of bioactive molecules. The gap hybridization strategy, the sandwich configuration, and the engineering aptamer strategy were employed for the assay of miRNA biomarkers, protein cancer biomarkers, and small molecules, respectively. The arrays showed good capability to anchor capture biomolecules for improving biorecognition. Addressable and high-throughput analysis with improved sensitivity and specificity had been achieved. The limit of detection for let-7a miRNA, prostate specific antigen, and cocaine were 10 fM, 40 pg/mL, and 100 nM, respectively. More importantly, we demonstrated that the microarray platform worked well with clinical serum samples and showed good relativity with conventional chemical luminescent immunoassay. We have developed a novel approach for the fabrication of DNA tetrahedron-based microarrays and a universal DNA tetrahedron-based microarray platform for the detection of different types of bioactive molecules. The microarray platform shows great potential for clinical diagnosis

Highly Stable Graphene-Based Nanocomposite (GO–PEI–Ag) with Broad-Spectrum, Long-Term Antimicrobial Activity and Antibiofilm Effects

Various silver nanoparticle (AgNP)-decorated graphene oxide (GO) nanocomposites (GO–Ag) have received increasing attention owing to their antimicrobial activity and biocompatibility; however, their aggregation in physiological solutions and the generally complex synthesis methods warrant improvement. This study aimed to synthesize a polyethyleneimine (PEI)-modified and AgNP-decorated GO nanocomposite (GO–PEI–Ag) through a facile approach through microwave irradiation without any extra reductants and surfactants; its antimicrobial activity was investigated on Gram-negative/-positive bacteria (including drug-resistant bacteria) and fungi. Compared with GO–Ag, GO–PEI–Ag acquired excellent stability in physiological solutions and electropositivity, showing substantially higher antimicrobial efficacy. Moreover, GO–PEI–Ag exhibited particularly excellent long-term effects, presenting no obvious decline in antimicrobial activity after 1 week storage in physiological saline and repeated use for three times and the lasting inhibition of bacterial growth in nutrient-rich culture medium. In contrast, GO–Ag exhibited a >60% decline in antimicrobial activity after storage. Importantly, GO–PEI–Ag effectively eliminated adhered bacteria, thereby preventing biofilm formation. The primary antimicrobial mechanisms of GO–PEI–Ag were evidenced as physical damage to the pathogen structure, causing cytoplasmic leakage. Hence, stable GO–PEI–Ag with robust, long-term antimicrobial activity holds promise in combating public-health threats posed by drug-resistant bacteria and biofilms

Gold Nanoparticle-Based Enzyme-Linked Antibody-Aptamer Sandwich Assay for Detection of <i>Salmonella</i> Typhimurium

Enzyme-linked immunosorbent assay (ELISA) provides a convenient means for the detection of <i>Salmonella enterica</i> serovar Typhimurium (STM), which is important for rapid diagnosis of foodborne pathogens. However, conventional ELISA is limited by antibody–antigen immunoreactions and suffers from poor sensitivity and tedious sample pretreatment. Therefore, development of novel ELISA remains challenging. Herein, we designed a comprehensive strategy for rapid, sensitive, and quantitative detection of STM with high specificity by gold nanoparticle-based enzyme-linked antibody-aptamer sandwich (nano-ELAAS) method. STM was captured and preconcentrated from samples with aptamer-modified magnetic particles, followed by binding with detector antibodies. Then nanoprobes carrying a large amount of reporter antibodies and horseradish peroxidase molecules were used for colorimetric signal amplification. Under the optimized reaction conditions, the nano-ELAAS assay had a quantitative detection range from 1 × 10³ to 1 × 10⁸ CFU mL^–1, a limit of detection of 1 × 10³ CFU mL^–1, and a selectivity of >10-fold for STM in samples containing other bacteria at higher concentration with an assay time less than 3 h. In addition, the developed nanoprobes were improved in terms of detection range and/or sensitivity when compared with two commercial enzyme-labeled antibody signal reporters. Finally, the nano-ELAAS method was demonstrated to work well in milk samples, a common source of STM contamination

Electrochemical DNA Biosensor Based on a Tetrahedral Nanostructure Probe for the Detection of Avian Influenza A (H7N9) Virus

A DNA tetrahedral nanostructure-based electrochemical biosensor was developed to detect avian influenza A (H7N9) virus through recognizing a fragment of the hemagglutinin gene sequence. The DNA tetrahedral probe was immobilized onto a gold electrode surface based on self-assembly between three thiolated nucleotide sequences and a longer nucleotide sequence containing complementary DNA to hybridize with the target single-stranded (ss)­DNA. The captured target sequence was hybridized with a biotinylated-ssDNA oligonucleotide as a detection probe, and then avidin-horseradish peroxidase was introduced to produce an amperometric signal through the interaction with 3,3′,5,5′-tetramethylbenzidine substrate. The target ssDNA was obtained by asymmetric polymerase chain reaction (PCR) of the cDNA template, reversely transcribed from the viral lysate of influenza A (H7N9) virus in throat swabs. The results showed that this electrochemical biosensor could specifically recognize the target DNA fragment of influenza A (H7N9) virus from other types of influenza viruses, such as influenza A (H1N1) and (H3N2) viruses, and even from single-base mismatches of oligonucleotides. Its detection limit could reach a magnitude of 100 fM for target nucleotide sequences. Moreover, the cycle number of the asymmetric PCR could be reduced below three with the electrochemical biosensor still distinguishing the target sequence from the negative control. To the best of our knowledge, this is the first report of the detection of target DNA from clinical samples using a tetrahedral DNA probe functionalized electrochemical biosensor. It displays that the DNA tetrahedra has a great potential application as a probe of the electrochemical biosensor to detect avian influenza A (H7N9) virus and other pathogens at the gene level, which will potentially aid the prevention and control of the disease caused by such pathogens

Nano Rolling-Circle Amplification for Enhanced SERS Hot Spots in Protein Microarray Analysis

Although “hot spots” have been proved to contribute to surface enhanced Raman scattering (SERS), less attention was paid to increase the number of the “hot spot” to directly enhance the Raman signals in bioanalytical systems. Here we report a new strategy based on nano rolling-circle amplification (nanoRCA) and nano hyperbranched rolling-circle amplification (nanoHRCA) to increase “hot spot” groups for protein microarrays. First, protein and ssDNA are coassembled on gold nanoparticles, making the assembled probe have both binding ability and hybridization ability. Second, the ssDNAs act as primers to initiate in situ RCA reaction to produced long ssDNAs. Third, a large number of SERS probes are loaded on the long ssDNA templetes, allowing thousands of SERS probes involved in each biomolecular recognition event. The strategy offered high-efficiency Raman enhancement and could detect less than 10 zeptomolar protein molecules in protein microarray analysis

Graphene Nanoprobes for Real-Time Monitoring of Isothermal Nucleic Acid Amplification

Isothermal amplification is an efficient way to amplify DNA with high accuracy; however, the real-time monitoring for quantification analysis mostly relied on expensive and precisely designed probes. In the present study, a graphene oxide (GO)-based nanoprobe was used to real-time monitor the isothermal amplification process. The interaction between GO and different DNA structures was systematically investigated, including single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), DNA 3-helix, and long rolling circle amplification (RCA) and hybridization chain reaction (HCR) products, which existed in one-, two-, and three-dimensional structures. It was found that the high rigid structures exhibited much lower affinity with GO than soft ssDNA, and generally the rigidity was dependent on the length of targets and the hybridization position with probe DNA. On the basis of these results, we successfully monitored HCR amplification process, RCA process, and the enzyme restriction of RCA products with GO nanoprobe; other applications including the detection of the assembly/disassembly of DNA 3-helix structures were also performed. Compared to the widely used end-point detection methods, the GO-based sensing platform is simple, sensitive, cost-effective, and especially in a real-time monitoring mode. We believe such studies can provide comprehensive understandings and evocation on design of GO-based biosensors for broad application in various fields

Multicolor Gold–Silver Nano-Mushrooms as Ready-to-Use SERS Probes for Ultrasensitive and Multiplex DNA/miRNA Detection

Uniform silver-containing metal nanostructures with strong and stable surface-enhanced Raman scattering (SERS) signals hold great promise for developing ultrasensitive probes for biodetection. Nevertheless, the direct synthesis of such ready-to-use nanoprobes remains extremely challenging. Herein we report a DNA-mediated gold–silver nanomushroom with interior nanogaps directly synthesized and used for multiplex and simultaneous SERS detection of various DNA and RNA targets. The DNA involved in the nanostructures can act as not only gap DNA (mediated DNA) but also probe DNA (hybridized DNA), and DNA’s involvement enables the nanostructures to have the inherent ability to recognize DNA and RNA targets. Importantly, we were the first to establish a new method for the generation of multicolor SERS probes using two different strategies. First Raman-labeled alkanethiol probe DNA was assembled on gold nanoparticles, and second, thiol-containing Raman reporters were coassembled with the probe DNA. The ready-to-use probes also give great potential to develop ultrasensitive detection methods for various biological molecules