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Emerging photocatalytic technology promises to provide an effective solution to the global energy crisis and environmental pollution. Graphite carbon nitride (g-C

Accurate, sensitive, and rapid detection of Salmonella and determination of whether it carries drug resistance genes plays an important role in guiding the clinical medication of salmonellosis and laying a foundation for studying the mechanism of drug resistance transmission of Salmonella. Here, a novel nontransferable, ultrasensitive dual detection platform (Cas12a-Ddp) was developed. The round cap allowed for temporary storage of more Cas12a detection solution than flat cap, enabling one-pot assays and reducing aerosol contamination. The results were read out in dual mode by the microplate reader and UV visualization to achieve sensitive dual-target detection of the virulence genes and drug resistance genes of Salmonella simultaneously, with the possibility of onsite detection. Cas12a-Ddp was combined with multiple polymerase chain reactions and recombinase polymerase amplifications successively. An ultrasensitive dual detection limit of 1 CFU/mL was obtained without any cross-reaction within 40 min. This was an improvement of 1–2 orders of magnitude over the existing methods. Cas12a-Ddp overcame the influence of proteins and fat in liquid matrix foods. It was used for the detection of drug-resistant Salmonella in milk and skim milk powder, also with the dual detection limit of 1 CFU/mL and spiked recovery of 68.58%–158.49%. It was also used for the analysis of Salmonella resistance rate analysis. The Cas12a-Ddp provided a reliable, fast, sensitive, and practical multi-CRISPR detection platform.

AFB

The toxicity evaluation suffers from the absence of suitable models capable of replicating in the co-cultured cell microenvironment and the function of specific tissues in vitro. Motivated by this urgent need, this study aimed to describe a novel three-dimensional (3D) liver spheroid model. The model consisted of a triple co-culture of HepG2, EA.hy 926, and LX-2. Subsequently, it was used for the toxicity evaluation of aflatoxin B1 (AFB1), and its advantages over the two-dimensional (2D) model and the mono-type cell spheroid model were assessed. This study examined the effects of AFB1 on cell viability, proliferation, mitochondria, oxidative stress, and cell membranes. The results revealed that AFB1 greatly affected 2D cell membranes and oxidative stress levels (0.01 μg/mL; 24 h), and could also significantly affect 2D cell viability, proliferation, and mitochondria levels (1 μg/mL; 24 h). On the contrary, 3D cells were less susceptible to AFB1. Combined with the analysis of gene expression, both metabolic activation (cytochrome P450; CYP450) and detoxification efficiency (drug-metabolizing enzymes) were found to be higher in 3D cells than in 2D cells. Moreover, 3D cells in triple co-culture outperformed mono-type cell spheroids. Therefore, the advanced 3D co-cultured spheroid model constructed in this study allowed us to more realistically simulate the microenvironment in vitro, and was a valuable and precise model to study mycotoxins.

Rapid, accurate, and safe screening of foodborne pathogenic bacteria is essential to effectively control and prevent outbreaks of foodborne illness. Fluorescent sensors constructed from carbon dots (CDs) and nanomaterial-based quenchers have provided an innovative method for screening of pathogenic bacteria. Herein, an ultrasensitive magnetic fluorescence aptasensor was designed for separation and detection of Staphylococcus aureus (S. aureus). Multicolor fluorescent CDs with a long fluorescent lifetime (6.73 ns) and high fluorescence stability were synthesized using a facile hydrothermal approach and modified cDNA as a highly sensitive fluorescent probe. CD fluorescence was quenched by Fe3O4 + aptamer via fluorescence resonance energy transfer (FRET). Under optimal conditions, the FRET-based aptasensor can detect S. aureus accompanied by a wide linear range of 50-107 CFU·mL-1 and a detection limit of 8 CFU·mL-1. Compared with other standard methods, this method was faster and more convenient, and the entire test was finished within 30 min. The capability of the aptasensor was simultaneously investigated on food samples. Additionally, the developed CDs exhibited excellent biocompatibility and were thus applied as fluorescent probes for bioimaging both in vitro and in vivo. This new platform provided an excellent application of the CDs for detecting and bioimaging pathogenic bacteria.

Ozone plays an increasingly important role in food processing for its antimicrobial ability and degradation effects on mycotoxins. The mycotoxin deoxynivalenol (DON) was treated with saturated aqueous ozone for different amounts of time, and by-products were collected for compounds annotation and cytotoxicity evaluation. To investigate the cytotoxicity of ozone degradation by-products, untargeted GC-TOFMS-based metabolomics were utilized. Caco-2 cells were dosed with 0.1 μg/mL DON and saturated aqueous ozone-treated DON (treatment time: 1 min, 3 min, 5 min) for 24 h followed by cytotoxicity tests (cell viability assay, ROS assay, and apoptosis assay), and intracellular metabolic analysis. Cytotoxicity test results revealed that ozone treatment could degrade DON structure; however, its degradation products and cellular toxicity existed under different treatment time of ozone. Metabolomics analysis indicated that ozone-treated DON degradation products weakened DON-induced metabolic disorder, such as purines-related nucleotide metabolism; Krebs cycle-related fuel and energy metabolism; and lipid, alkaloid and amino acid metabolism. By contrast, the catecholamine pathway, which is related to latent inflammation and oxidative stress effects, was unaltered in the ozone-treated DON group, indicating that the potential cytotoxicity still existed. These findings provide a comprehensive safety evaluation for ozone-treated DON in vitro and propose a new strategy for studying the effects of ozone-treated food.

Metabolic profiling in Caco-2 cells was studied for the combined toxic effects of deoxynivalenol (DON), zearalenone (ZEN), and Aflatoxin B1 (AFB1) through untargeted GC-MS. The GC-MS spectra of Caco-2 cells treated with individual 6.7 μM DON, 20 μM ZEN, 20 μM AFB1 and the combined DON + AFB1 (6.7 + 20 μM) and DON + ZEN + AFB1 (6.7 + 20 + 20 μM) for 24 h were deconvoluted, aligned and identified with MS DIAL. The metabolic pathway analysis was analyzed with MetaMapp and visualized with CytoScape. Results show that the combined DON + AFB1 and DON + ZEN + AFB1 treatment has an obvious “synergistic effect”. The apoptosis-related gene mRNA test result indicates that the combined mycotoxins downregulate Bcl-2 gene and upregulate Bax, p53, caspase-3, caspase-8 and caspase-utilized 9 genes, more significantly than any individual mycotoxins group. The cellular metabolism illustrated that the combined mycotoxin groups, DON + ZEN + AFB1 seriously effect glycine, serine and threonine metabolism, pyruvate metabolism, etc. while no metabolic disorders were presented in individual mycotoxin group. Our hypothesis was validated that the combined mycotoxins with low concentrations can have a synergistic effect in the metabolism, which may lead to cellular apoptosis or necrosis.

An efficient strategy based on the nanosurface energy transfer (NSET) between gold nanocrosses (AuNCs) and graphene quantum dots (GQDs) for intracellular ATP sensing and tracking was first developed. GQDs and AuNCs assembled in the bases of complementary matching principle, which result in the fluorescence quenching via NSET. In the presence of ATP, aptamer recognized and combined with ATP, caused the distance change between GQDs and AuNCs. The fluorescence intensity was proportional to ATP concentration, which was represented by the equation y = 176.67 x + 335.51 (R2 = 0.9916) with the detection limit of 0.27 mM (3δ, n = 11). Moreover, the proposed “OFF-ON” sensor showed low cytotoxicity and excellent performance in intracellular ATP sensing, which will provide new possibilities for labeling of small biological molecules.

A novel electrochemical sensor for paraquat (PQ) detection based on molecularly imprinted polymer (MIP) membranes on a glassy carbon electrode (GCE) modified with Au nanocrosses-chitosan (AuNCs-CS) was constructed. P-Aminothiophenol (p-ATP) and 4,4′-bipyridine template were assembled on the surface of the modified GCE by the formation of Au–S bonds and hydrogen-bonding interactions, followed by polymer membrane formation by the electropolymerization in a polymer solution containing p-ATP, HAuCl4, tetrabutylammonium perchlorate (TBAP), and the template molecule 4,4′-bipyridine. The as-constructed molecularly imprinted sensor (MIP-AuNC-CS) was characterized by differential pulse voltammetry (DPV), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). This is the first time that molecularly imprinted polymer technology has been integrated with a modified AuNCs-CS to electrochemically detect PQ. The linear response of the MIP-AuNC-CS sensor was in the range from 1 × 10−14 to 1 × 10−10 mol L−1, and the limit of detection was 2.3 × 10−15 mol L−1. This sensor showed high-speed real-time detection capability, low sample consumption, high sensitivity, low interference, and good stability characteristics, and was proven to detect PQ.

Food quality and safety has become a subject of major concern for authorities and professionals in the food supply chain. Rapid methods, particularly biosensors, have exceptional specificity and sensitivity, rapid response times, low cost, relatively compact size, and are user friendly to operate. Cell-based biosensors are portable, and provide the biological activity of the analyte suitable for an initial screening of food. In this overview, the utilization of cell-based biosensors for food safety and quality analyses, such as detecting toxins, foodborne pathogens, allergens, and evaluating toxicity and function are summarized. Our results will promote the future development of cell-based biosensors in the food field.

We describe a “turn-on” magnetic fluorescent biosensor based on graphene quantum dots (GQDs), Fe3O4, and molybdenum disulfide (MoS2) nanosheets. It is used for rapid, efficient, and sensitive separation and detection of circulating tumor cells (CTCs). A facile approach (electrochemical synthesis method) for the preparation of photoluminescent GQDs functionalized with an aptamer [epithelial cell adhesion molecule (EpCAM) receptors] and a magnetic agent for one-step bioimaging and enrichment of CTCs is described. MoS2 nanosheets, as a fluorescence quencher, and the aforementioned aptamer@Fe3O4@GQD complex were assembled to construct “turn-on” biosensing magnetic fluorescent nanocomposites (MFNs). This system exhibits low cytotoxicity and an average capture efficiency of 90%, which is higher than that of other magnetic nanoparticles on account of the one-step CTC separation method. In addition, the MFNs could quickly identify and label CTCs within 15 min, surpassing other one-step and two-step marker detection methods. Furthermore, because of the presence of aptamers, the MFNs have specific capability to capture CTCs (both low- and high-EpCAM-expressing cells). In addition, high-sensitivity detection of up to ten tumor cells in whole blood was achieved. Therefore, the MFNs have great potential to be used as universal biosensing nanocomposites for fluorescence-guided tumor cell enrichment and bioimaging.

Microcystins (MCs) are a class of toxins that are mainly produced by cyanobacteria. Among them, microcystin-leucine arginine (MC-LR) is one of the most toxic and harmful of the freshwater toxins, and it has caused many accidents and threats to human health. In the present study, we report a neoteric lateral flow fluorescent microsphere immunochromatographic assay (FM-ICA) combined with UV light detection to rapidly and quantitatively detect MC-LR in freshwater food, based on direct competitive immunoreactions on chromatographic strips. In the method, MC-LR artificial antigen, MC-LR-bovine serum albumin (MC-LR-BSA), and goat anti-rabbit immunoglobulin G (IgG) were labelled with europium (Eu) nanospheres that functioned as a luminescence tracer. Goat anti-mouse IgG and rabbit IgG were spread on a nitrocellulose (NC) membrane for the test (T) line and control (C) line, respectively. The optimal parameters were 1 mg/mL for the goat anti-mouse IgG and rabbit IgG, 20 μg of MC-LR-BSA conjugated with Eu nanospheres, 1:100 for the dilution of Eu-MC-LR-BSA, and a 1:30000 dilution of anti-MC-LR monoclonal antibody (mAb) with an assay time of 15 min. The working range of the MC-LR standard curve was 0.1–5.0 μg/L with a limit of detection (LOD) of 0.0542 μg/L and a 50% inhibitory concentration (IC50) of 0.5613 μg/L. The average recoveries of the FM-ICA for detecting MC-LR in real samples ranged from 90.1 to 109.3%, with coefficients of variation under 15.8%. The FM-ICA test strips for quantitative determination of MC-LR provide sensitive, simple, and rapid performance, and can be used to monitor freshwater samples.

A graphene modified glassy carbon electrode was fabricated for sensitive and convenient electrochemical detection of 1-hydroxypyrene (1-OHP). The graphene modified electrode was prepared by eletrochemical reduction of graphene oxide on glass carbon electrode. l-OHP was synergistically accumulated on the graphene modified electrode via π-π conjugate adsorption and electrochemical oxidation. Electrochemical behavior of l-OHP on the graphene modified glass carbon electrode was studied in detail. The developed method gave a linear calibration curve for the determination of 1-OHP in the range of 5–300 nM (R2 = 0.997), a limit of detection of 0.84 nM (S/N = 3). The proposed method was also successfully applied to the analysis of urine samples for l-OHP with the recoveries of 97.3–101.1%.

Fumonisin B1 (FB1) is a neurodegenerative mycotoxin synthesized by Fusarium spp., but the potential neurobehavioral toxicity effects in organisms have not been characterized clearly. Caenorhabditis elegans (C. elegans) has emerged as a promising model organism for neurotoxicological studies due to characteristics such as well-functioning nervous system and rich behavioral phenotypes. To investigate whether FB1 has neurobehavioral toxicity effects on C. elegans, the motor behavior, neuronal structure, neurotransmitter content, and gene expression related with neurotransmission of C. elegans were determined after exposed to 20-200 μg/mL FB1 for 24 h and 48 h, respectively. Results showed that FB1 caused behavioral defects, including body bends, head thrashes, crawling distance, mean speed, mean amplitude, mean wavelength, foraging behavior, and chemotaxis learning ability in a dose-, and time-dependent manner. In addition, when C. elegans was exposed to FB1 at a concentration of 200 μg/mL for 24 h and above 100 μg/mL for 48 h, the GABAergic and serotonergic neurons were damaged, but no effect on dopaminergic, glutamatergic, and cholinergic neurons. The relative content of GABA and serotonin decreased significantly. Furthermore, abnormal expression of mRNA levels associated with GABA and serotonin were found in nematodes treated with FB1, such as unc-30, unc-47, unc-49, exp-1, mod-5, cat-1, and tph-1. The neurobehavioral toxicity effect of FB1 may be mediated by abnormal neurotransmission of GABA and serotonin. This study provides useful information for understanding the neurotoxicity of FB1.

In this study, the possible molecular mechanisms of zearalenone (ZEA)-induced reproductive and developmental toxic effects in Caenorhabditis elegans (C. elegans) were investigated. Differential gene expression profiles were identified, and 171, 245, and 3149 genes were down- or up-regulated (>2.0 fold) in 10, 20, and 40 μg/ml ZEA treated groups, respectively, as compared to untreated controls. Pathway specific mapping showed that the major differentially expressed genes were collagen synthetic pathways regulating genes, col-121 and dpy-17. Real-time PCR reconfirmation of key genes, related to cuticle collagen synthetic pathway, found dramatic changes in the expression of the genes dpy-31, sqt-3, col-121, and dpy-17 following exposure to ZEA (40 μg/ml), which indicated the significance of these genes in ZEA-induced toxicity. Cuticle collagen plays many key roles in the development and reproduction of C. elegans. The hypersensitive responses in transgenic and mutant worms also confirmed the roles of these genes in lethality and reproductive response to ZEA exposure, which indicates that ZEA blocked the normal collagen processing and cuticle formation. Taken together, our results demonstrate that disruption of the collagen biosynthetic pathway might be a key mechanism in ZEA-induced reproductive and developmental toxic effects in C. elegans.

Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that contaminate a wide range of grains and crops. In this study, a one-step time-resolved single-channel immunochromatographic test strip based on europium ion polystyrene fluorescence microspheres was first developed for sensitive and quantitative detection of DON and ZEN. The concentration of the artificial antigen and the mass ratio of the monoclonal antibody to fluorescent microspheres for conjugation were optimized to simplify the sample addition process during immunochromatographic assay and improve the on-site detection efficiency. The limits of detection (LOD) of the single-channel immunochromatographic test strip for DON and ZEN detection were 0.17 and 0.54 μg/L, respectively. Meanwhile, the dual-channel immunochromatographic test strip was designed to simultaneously detect DON and ZEN, with LODs of 0.24 and 0.69 μg/L achieved for DON and ZEN, respectively. The developed test strips also yielded recovery results consistent with that obtained by LC-MS/MS for DON and ZEN detection in real samples of wheat and corn flour, confirming the practicability and reliability of the test strip. The developed immunochromatographic test strips realize quick and sensitive detection of DON and ZEN, exhibiting potential for broad applications in the point-of-care testing platform of multiple mycotoxins in agricultural products. Graphic abstract.

Methamphetamine (METH) abuse has become a global public health problem. However, the potential mechanisms involving METH-induced metabolic disorders have thus far remained poorly understood. Metabolomics can provide a clue for the cause of apparent changes and consequently be used to investigate the METH-induced dysregulation of metabolite expression and the mechanism of metabolic disorder mechanism. This laboratory investigation included 80 METH abusers and 80 healthy people. The serum metabolites were detected and analysed by gas chromatography/time-of-flight mass spectrometry. Raw data were processed with the software MS DIAL, which includes deconvolution, peak alignment and compound identification. The data matrix was processed by univariate and multivariate analyses for significant metabolite screening with the criteria of variable importance in projection values > 1, fold change > 1.5 and the t test (p value < 0.05). Significant differences in 16 metabolites (deoxycholic acid, cholic acid, hydroxylamine, etc.) in serum were found between the METH abuse group and the control group. Energy metabolic pathways and several amino acid metabolic pathways (alanine, aspartic acid and glutamate metabolism and tryptophan metabolism) were primarily involved. Further analysis indicated that the area under the receiver operating characteristic curve (AUC) was 0.998 for these 16 metabolites. Among the metabolites, three carbohydrates (d-ribose, cellobiose and maltotriose) had an AUC of 0.975, which were determined as potential markers of abuse. We observed metabolic disturbances in METH abusers, particularly perturbation in energy metabolism and amino acid metabolism, which can provide new insights into the search for biomarkers and the mechanisms underlying the adverse effects of METH on human health.

Mycotoxins are metabolites produced by fungi growing in food or feed, which can produce toxic effects and seriously threaten the health of humans and animals. Mycotoxins are commonly found in food and feed, and are of significant concern due to their hepatotoxicity, nephrotoxicity, carcinogenicity, mutagenicity, and ability to damage the immune and reproductive systems. Traditional physical and chemical detoxification methods to treat mycotoxins in food and feed products have limitations, such as loss of nutrients, reagent residues, and secondary pollution to the environment. Thus, there is an urgent need for new detoxification methods to effectively control mycotoxins and treat mycotoxin pollution. In recent years, microbial detoxification technology has been widely used for the degradation of mycotoxins in food and feed because this approach offers the potential for treatment with high efficiency, low toxicity, and strong specificity, without damage to nutrients. This article reviews the application of microbial detoxification technology for removal of common mycotoxins such as Aflatoxin, Ochratoxin, Zearalenone, Deoxynivalenol, and Fumonisins, and discusses the development trend of this important technology.

There are many kinds of estrogens, and endogenous estrogens produce a variety of estrogen metabolites with similar structure but with different physiological effects after metabolism in vivo. Studies have shown that estrone (E1) widely occurs in the environment and animal-derived food. Because of its estrogen effect, E1 can have adverse effects on the human body as an endocrine disruptor. In this study, we found that E1 and 2-hydroxyestrone (2-OH-E1), the hydroxylation metabolite of estrogen, have opposite proliferative effects on breast cancer cells (MCF-7) through cell proliferation experiments and comparison of their effects by molecular docking and detection of ROS, Ca2+, and cell pathway proteins. The effects of 2-methoxyestrone (2-MeO-E1) and 16α-hydroxyestrone (16α-OH-E1) on the biochemical and protein levels of MCF-7 were further studied to compare the effects of metabolic sites and modes on estrogen effects. Hydroxylation of E1 at the C2 site weakened the estrogen effect, down-regulated the expression of the mammalian target of rapamycin (mTOR) and protein kinase B (Akt) pathway proteins, inhibited the proliferation of cancer cells, and enhanced anti-oxidative stress and anti-inflammation. Methoxylation at the C2 position also inhibited the expression of inflammatory and oxidative stress pathway proteins but did not greatly affect the estrogen effects. However, hydroxylation on C16 had no significant effect on the biological effects of estrogen. Therefore, the structural changes of estrogen on C2 are important reasons for the different physiological effects of estrogen and its metabolites. Thus, by regulating the gene Cytochrome P450 1B1(CYP1B1), which affects the hydroxylation metabolism of estrogen, and promoting the hydroxylation of estrone at the C2 position, the estrogen effect of estrone can be effectively reduced, thus reducing the harm its poses in food and the environment.

Single-cell electrochemical sensor is widely used in the local selective detection of single living cells because of its high spatial-temporal resolution and sensitivity, as well as its ability to obtain comprehensive cellular physiological states and processes with increased accuracy. Functionalized nanoprobes can detect the oxidative stress response of cells in single-cell electrochemical sensors. Moreover, the T-2 toxin is one of the most toxic mycotoxins and widely occurs in field crops. T-2 toxin can cause mitochondrial damage in cells and increase intracellular reactive oxygen species (ROS) in various cells. As the most representative free radical of intracellular ROS, H2O2 can effectively reflect the toxic effects of intracellular T-2 toxin. In this study, a functionalized gold nanoprobe was used to dynamically monitor the production of H2O2 in a single live human hepatoma cell HepG2 stimulated by mycotoxin T-2. The concentration of H2O2 produced by HepG2 cells stimulated by T-2 toxin at 1 ppb-1 ppm was linearly correlated, R2 = 0.99055, and LOD = 0.13807 ng mL-1. Sample spiking experiments were conducted, and the recovery rate of spiking was 81.19%-130.17%. A comparative analysis of differences in the current produced by multiple toxins, HT-29 cells, as well as single cells in cell populations, was performed. This method can be applied in real-time monitoring of mycotoxin toxicity during food processing in living cells and provides a novel idea for enhancing food quality and safety in a nanoenvironment.

A new, green, and cost-effective magnetic solid-phase extraction of aflatoxins and ochratoxins from edible vegetable oils samples was developed using polydopamine-coated magnetic multi-walled carbon nanotubes (PDA@Fe3O4-MWCNTs) as the absorbent. PDA@Fe3O4-MWCNTs nanomaterials were prepared by chemical co-precipitation and in situ oxidation and self-polymerization of dopamine and was characterized. Factors affecting MSPE and the adsorption behavior of the adsorbent to mycotoxins were studied, and the optimal extraction conditions of MSPE and the complexity of the adsorption process were determined. Based on this, the magnetic solid-phase extraction-high-performance liquid chromatography-fluorescence detection method (MSPE-HPLC-FLD) was established for determining six mycotoxins [aflatoxin B1 (AFB1), AFB2, AFG1, and AFG2, and ochratoxin A (OTA) and OTB)] in vegetable oils. The recovery was 70.15%~89.25%, and RSD was ≤6.4%. PDA@Fe3O4-MWCNTs showed a high affinity toward aflatoxins and ochratoxins, allowing selective extraction and quantification of aflatoxins and ochratoxins from complex sample matrices.

The coexistence of mycotoxins in agricultural products poses a serious threat to food safety. This study developed a dual immunochromatographic test strips (DICTS) method based on double antibodies labeled with time-resolved fluorescent microspheres (TRFM) to realize simultaneous rapid detection of aflatoxin M1 (AFM1) and ochratoxin A (OTA) in milk. As bridge antibody, the polyclonal antibody (pAb) was first conjugated with the TRFM and then with the monoclonal antibody (mAb). Meanwhile, a biotin-streptavidin system was introduced to replace the traditional goat anti-mouse Immunoglobulin G, thus providing a stable signal on the control line. After optimization, the detection limit of AFM1 and OTA in milk was respectively 0.018 and 0.036 ng/mL. The recoveries of intraassay and interassay experiments ranged from 89.65% to 103.99%. The accuracy, repeatability, and specificity of the developed TRFM-DICTS were estimated. The results of TRFM-DICTS showed a high consistency with those of the ultrahigh-performance liquid chromatography-tandem mass spectrometry.

AFB1 and heavy metal Cd are two common pollutants during grain storage. The rapid detection of grains before they enter the granary is particularly important. Hence, rapidly, accurately, and sensitively screening contaminated grains, simplifying the detection process, and reducing detection costs are necessary. In this study, linear ranges of time-resolved fluorescence microsphere−immunochromatographic test strip (TRFM-ICTS) detection were 0.01–30 ng/mL (AFB1) and 0.01–60 ng/mL (Cd), and the IC50 values were 0.536 ng/mL (AFB1) and 3.331 ng/mL (Cd). In the TRFM-ICTS sample addition experiment, the recovery rates were all between 90% and 110%. The coefficient of variation was less than 8% in the actual sample detection process of grain. We have established a one-step extraction method for AFB1 and Cd in grains to achieve simultaneous detection in one extraction. In addition, TRFM-ICTS could be stored for at least 12 months, providing technical support for the realization of commercial production.

A “honeycomb” electrochemical biosensor based on 3D printing was developed to noninvasively monitor the viability of 3D cells and evaluate the individual or combined toxicity of deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), and 15-acetyldeoxynivalenol (15-ADON). Carbon nanofiber (CN)/gelatin methacryloyl (GelMA) conductive composite hydrogel with strong processability was printed on 8-channel screen-printed carbon electrodes (SPCEs) to maintain cell viability and form tight cell-to-cell contacts. A “3D honeycomb” printing infill pattern was selected in the construction of the biosensors to improve conductivity. Based on 3D printing technology, the electrochemical biosensor can prevent manual error and provide for high-throughput detection. Electrochemical impedance spectroscopy (EIS) was used to evaluate mycotoxin toxicity. The EIS response decreased with the concentration of DON, 3-ADON and 15-ADON in the range of 0.1–10, 0.05–100, and 0.1–10 μg/mL, respectively, with a limit of detection of 0.07, 0.10 and 0.06 μg/mL, respectively. Mycotoxin interactions were analyzed using the isobologram–combination index (CI) method. The electrochemical cytotoxicity evaluation result was confirmed by biological assays. Therefore, a novel method for evaluating the combined toxicity of mycotoxins is proposed, which exhibits potential for application to food safety and evaluation.

Antibiotic resistance has become a serious threat to food safety and public health globally. Therefore, the development of a sensitive, quick, and simple method for antibiotic susceptibility testing is an urgent and crucial need. A novel concentration gradient microfluidic chip was designed in this work to generate antibiotic concentration gradient, culture bacteria, and produce fluorescence emission. An in-house-assembled fluorescence detection platform was constructed, and experiments were conducted to verify the linearity of the generated concentration gradient, explore the appropriate incubation time and flow rate for the microfluidic chip, and study the effect of long-term acid-based food processing on antibiotic susceptibility testing. Experimental results show that the concentration gradient generated by the microfluidic chip exhibited good linearity, stability, and controllability. The appropriate flow rate and incubation time for the microfluidic chip were 2 μL/min and 5 h, respectively. The use of this microfluidic chip for testing antibiotic resistance of Salmonella to ofloxacin and ampicillin generated results that were completely consistent with test results obtained using the gold-standard method. Furthermore, Salmonella showed greater sensitivity to antibiotics under strong acid conditions, confirming the potential influence of acid-based food processing on antibiotic susceptibility testing of real samples. The designed microfluidic chip provides a high-throughput, sensitive, and rapid antibiotic susceptibility testing method that combines the microfluidic chip and the fluorescence detection platform. The application of this method would facilitate determination of antibiotic-resistant bacterial strains for clinicians and researchers, and enable monitoring of changes in bacterial resistance during food processing.

Foodborne diseases caused by pathogens and toxins are a serious threat to food safety and human health; thus, they are major concern to society. Existing conventional foodborne pathogen or toxin detection methods, including microbiological assay, nucleic acid-based assays, immunological assays, and instrumental analytical method, are time-consuming, labor-intensive and expensive. Because of the fast response and high sensitivity, cell-based biosensors are promising novel tools for food safety risk assessment and monitoring. This review focuses on the properties of mammalian cell-based biosensors and applications in the detection of foodborne pathogens (bacteria and viruses) and toxins (bacterial toxins, mycotoxins and marine toxins). We discuss mammalian cell adhesion and how it is involved in the establishment of 3D cell culture models for mammalian cell-based biosensors, as well as evaluate their limitations for commercialization and further development prospects.

Antibacterial hydrogels have received attention for preventing infections and for their biomedical applications. However, traditional antibiotics-containing and metal nanoparticle-containing hydrogels often cause bacterial resistance, exhibit low biocompatibility, and lack real-time monitoring capability. Here, a fluorescent antibacterial hydrogel with antibacterial ability, excellent optical performance, and high biocompatibility was developed based on cationic carbon dots (CDs), pectin, and acrylic acid triggered construction of the hydrogel network by cross-linker. The antibacterial high-cationic CDs (+51.20 mV) were synthesized by a simple hydrothermal method and released from hydrogel in response to broken hydrogen bonds due to a change in the ambient environment caused by the growing bacteria. The hydrogel showed long-term potent broad-spectrum antibacterial ability (even drug-resistant bacteria) due to the bacterial membrane seriously damaged by the released CDs. The inhibitory capability of this hydrogel was 108.5-fold higher than the other hydrogel. After implantation or incubation with cells, no obvious cytotoxicity or tissue toxicity was observed for the antibacterial hydrogel. This hydrogel enhanced both the application of CDs in vivo and the biosafety of hydrogel. Furthermore, the multicolor fluorescence emission produced by CD provides a potential idea for the development of dual-function hydrogels with in situ monitoring and prevention of bacterial infections to treat wounds.

Paraquat (PQ) residue is harmful for human health, agriculture, and the aquatic environment. This paper proposes a novel fluorescent molecularly imprinted polymer (MIP), SiO

The pollution caused by estrogens in the environment and food has received increasing attention. It is still challenging for on-site immunochromatographic assay (ICA) detection of estrogens. The performance of the prepared probes plays a decisive role in the sensitivity and stability of the ICA system. The published probes usually directly couple the detection antibody to the label, ignoring the influence of the label on the activity of the antibody. In this study, 17β-estradiol (E2) was used as a model analyte for the ICA system. Two universal probes were constructed based on quantum dot nanobeads (QBs), recombinant protein A (SPA, from Staphylococcus aureus), and rabbit anti-mouse immunoglobulin G antibody (anti-IgG). The probes were prepared by coupling QBs with SPA, releasing anti-E2 monoclonal antibody (mAb), and maintaining its activity. The prepared universal probes can orient recognize the Fc region of mAb and fully expose its Fab region, improving the detection sensitivity of the ICA system. The free anti-E2 mAb and the universal probe (QBs@SPA or QBs@SPA@anti-IgG) were used as the detection antibodies and signal donors, respectively. The results show that the proposed ICA based on QBs@SPA and QBs@SPA@anti-IgG probes could detect E2 with IC50 of 8.83 and 0.93 ng/mL, respectively, within 15 min under optimal conditions. The recovery results of ICA based on QBs@SPA and QBs@SPA@anti-IgG probes showed good agreement with the findings of the high-performance liquid chromatography (HPLC) analysis for spiked samples. The developed ICA system based on universal probes was superior in terms of sensitivity, rapidity, and applicability, and held great promise for its implementation in detecting environmental and food small-molecule pollutants.

In this study, a fluorine-fluorine interaction approach through fluoridating boron nitride nanosheets (BNNs) for sensing perfluoroalkyl acids (PFAAs) in multiple food matrices was developed. Through a facile hydrothermal fluorination modification, the BNNs were transferred into homogeneous fluorinated boron nitride nanoparticles (F-BNNs) with robust networks and specific surface area. After morphological modification, the particles displayed strong adsorption and sensing capabilities on PFAAs in both solid and liquid food matrix. Under the evaluation of mass spectrometry, F-BNNs based microextraction approach exhibited low method detection limits (MDLs) in the ranges of 0.9–3.9 pg mL−1 and 3.6–15.8 pg g−1 for milk and meat matrices, respectively, with satisfactory repeatability (RSD%

Antioxidant evaluation of bioactive compounds is limited, since many methods lack a real physiological environment that can be used conveniently and intuitively. In this study, a simple, label-free and effective electrochemical biosensor method has been developed to evaluate the antioxidant effect of phloretin (Ph) by 3D cell modification on a glassy carbon electrode (GCE). In response to this, A549 cells were immobilized onto a self-assembled ʟ-cysteine/gold nanoparticle (AuNPs/ʟ-Cys)-modified GCE surface by a simple drop casting after encapsulated in alginate. The electrochemical impedance spectroscopy (EIS) results showed that the impedance value (Ret) increased with the concentration of H2O2 in the range of 0–60 μmol/L with the correlation of 0.990 which acted as an oxidative stress model inducer. However, the EIS value decreased with the co-incubation of Ph ranging from 10 to 100 μmol/L, showing a dose-dependent manner and time effect, indicating that the variation of Ret was responded to the antioxidant effect. The response impedance of the biosensor is linear to Ph concentrations from 20 μmol/L to 100 μmol/L with the detection limit (LOD) as 1.96 μmol/L. A significant correlation was observed between reactive oxygen species (ROS) values and Ret values following the concentrations of Ph, thus demonstrating the good biological relevance of cell-based electrochemical method. The strategy has been used to evaluate Ph antioxidant capacity in real cells with satisfactory results, indicating the feasibility of biosensor analysis for antioxidant evaluation.

Enrofloxacin (ENR) is a widely used fluoroquinolone (FQ) antibiotic for antibacterial treatment of edible animal. In this study, a rapid and highly specific fluorescence polarization immunoassay (FPIA) was developed for monitoring ENR residues in animal foods. First, ENR was covalently coupled to bovine serum albumin (BSA) to produce specific polyclonal antibodies (pAbs). Three fluorescein-labeled ENR tracers (A, B, and C) with different spacers were synthesized and compared to obtain higher sensitivity. Tracer C with the longest arm showed the best sensitivity among the three tracers. The developed FPIA method showed an IC50 (50% inhibitory concentration) of 21.49 ng&middot, mL&minus, 1 with a dynamic working range (IC20&ndash, IC80) of 4.30&ndash, 107.46 ng&middot, 1 and a limit of detection (LOD, IC10) of 1.68 ng&middot, 1. The cross-reactivity (CR) of several structurally related compounds was less than 2%. The recoveries of spiked pork liver and chicken samples varied from 91.3% to 112.9%, and the average coefficients of variation were less than 3.83% and 5.13%, respectively. The immunoassay took only 8 min excluding sample pretreatment. This indicated that the established method had high sensitivity, specificity, and the advantages of simplicity. Therefore, the proposed FPIA provided a useful screening method for the rapid detection of ENR residues in pork liver and chicken.

Lipopolysaccharide (LPS), a characteristic component of the outer membrane of Gram-negative bacteria, can be used as an effective biomarker to detect bacterial contamination. Here, we reported a 293/hTLR4A-MD2-CD14 cell-based fluorescent biosensor to detect and identify LPS, which is carried out in a 96-well microplate which is nondestructive, user-friendly, and highly efficient. The promoter sequence of the critical signaling pathway gene ZC3H12A (encoding MCPIP1 protein) and enhanced green fluorescence protein (EGFP) were combined to construct a recombinant plasmid, which was transferred into 293/hTLR4A-MD2-CD14 cells through lipid-mediated, DNA-transfection way. LPS was able to bind to TLR4 and coreceptors-induced signaling pathway could result in green fluorescent protein expression. Results show that stable transfected 293/hTLR4A-MD2-CD14 cells with LPS treatment could be directly and continually observed under a high content screening imaging system. The novel cell-based biosensor detects LPS at low concentration, along with the detection limit of 0.075 μg/mL. The cell-based biosensor was evaluated by differentiating Gram-negative and Gram-positive bacteria and detecting LPS in fruit juices as well. This proposed fluorescent biosensor has potential in sensing LPS optically in foodstuff and biological products, as well as bacteria identification, contributing to the control of foodborne diseases and ensurance of public food safety with its high throughput detection way.

This review considers the evolution of optical-based biosensors for bacterial pathogen detection as the causative agents for different diseases. Conventional identification and detection techniques...

Near-infrared (NIR) spectroscopy as a tool for direct and quantitatively screening the minute polymorphic transitions of bioactive fatty acids was assessed basing on a thermal heating process of oleic acid. Temperature-dependent NIR spectral profiles indicate that dynamical variances of COOH group dominate its γ → α phase transition, while the transition from active α to β phase mainly relates to the conformational transfer of acyl chain. Through operating multivariate curve resolution-alternating least squares with factor analysis, instantaneous contribution of each active polymorph during the transition process was illustrated for displaying the progressive evolutions of functional groups. Calculated contributions reveal that the α phase of oleic acid initially is present at around −18 °C, but sharply grows up around −2.2 °C from the transformation of γ phase and finally disappears at the melting point. On the other hand, the β phase of oleic acid is sole self-generation after melt even it embryonically appears at −2.2 °C. Such mathematical approach based on NIR spectroscopy and factor analysis calculation provides a volatile strategy in quantitatively exploring the transition processes of bioactive fatty acids; meanwhile, it maintains promising possibility for instantaneous quantifying each active polymorph of lipid materials.

Lipopolysaccharide (LPS) can lead to uncontrollable cytokine production, fatal sepsis syndrome and depression/multiple organ failure, as pathophysiologic demonstration. Various toxic effects of LPS have been extensively reported, mainly on the toxicity of LPS in cellular level, macrophages or tumor cells, etc. This work aimed on the impact of LPS on mast cell metabolism, which focused on LPS-induced cellular metabolic profiles. Gas chromatography-mass spectrometry (GC-MS) based metabolomics strategy was implemented for the endo-metabolites detection in rat basophilic leukemia (RBL-2H3) cells, treated with 10 μg/mL LPS for 24 h, along with multiple time-dose tests of cells viability/apoptosis. Significantly changes metabolites were mainly involved the metabolism of glycine, serine, threonine and the biosynthesis of phenylalanine, tyrosine, tryptophan and pentose phosphate pathway. The endo-metabolism results illustrated that LPS treatment led to downregulation of glycine, serine and threonine metabolism besides pentose phosphate pathway in RBL-2H3 cells. This novel insight into LPS cellular metabolism, provides some heuristic guidance for elucidating the underlying mechanism of LPS-mediated disease.

Cyclocarya paliurus (C. paliurus) is a well-known new food resource whose extraction has certain antidiabetic activity. In this study, compounds were prepared from C. paliurus and their antidiabetic activity were further evaluated in vivo and in vitro. Besides, chemical structures of these compounds were analyzed using nuclear magnetic resonance (NMR). The study showed that compounds extracted from C. paliurus showed excellent antidiabetic activity, which was shown in hypoglycemic effect, lipid metabolism regulation, antioxidant enzyme system improvement, insulin sensitivity increasing and repairment of islet cells. Two major antidiabetic compounds were identified, namely quercetin-3-O-β-D-glucuronide-6′-methyl ester (compound 1) and kaempferol-3-O-β-D-glucuronide-6′-methyl ester (compound 2), and compound 1 was firstly discovered in C. paliurus. Compounds 1 and 2 were proved great inhibition in vitro with IC50 values of 48.25 μg/mL, 66.30 μg/mL of α-amylase inhibition and 115.2 μg/mL, 136.8 μg/mL of α-glucosidase inhibition, respectively. Molecular docking analysis proved that compounds can bind to the active sites of enzymes. The results explored the antidiabetic compounds of C. paliurus, which can further help promote development of C. paliurus and enlarge kinds of therapeutic alternatives to antidiabetic recipes.

Emerging photocatalytic technology promises to provide an effective solution to the global energy crisis and environmental pollution. Graphite carbon nitride (g-C3 N4 ) has gained extensive attention in the scientific community due to its excellent physical and chemical properties, attractive electronic band structure, and low cost. In this paper, research progress in design strategies for g-C3 N4 -based photocatalysts in the past five years is reviewed from the perspectives of nanostructure construction, element doping, and heterostructure construction. To clarify the relationship between application requirements and structural design, variations in the morphology, electronic energy band structure, light absorption capacity, as well as interfacial charge transfer caused by various modification strategies are discussed in detail. The recent applications of g-C3 N4 -based photocatalysts for pollutant degradation and bacterial disinfection are reviewed, as well as the antimicrobial activity and degradation mechanisms. Finally, current challenges and future development directions for the practical application of g-C3 N4 -based photocatalysts are tentatively discussed.

Traditional cell biology researches on cell populations by their origin, tissue, morphology, and secretions. Because of the heterogeneity of cells, research at the single-cell level can obtain more accurate and comprehensive information that reflects the physiological state and process of the cell, increasing the significance of single-cell analysis. The application of single-cell analysis is faced with the problem of contaminated or damaged cells caused by cell sample transportation. Reversible encapsulation of a single cell can protect cells from the external environment and open the encapsulation shell to release cells, thus preserving cell integrity and improving extraction efficiency of analytes. Meanwhile, microfluidic single cell analysis (MSCA) exhibits integration, miniaturization, and high throughput, which can considerably improve the efficiency of single-cell analysis. The researches on single-cell reversible encapsulation materials, single-cell analysis methods, and the MSCA integration platform are analyzed and summarized in this review. The problems of single-cell viability, network of single-cell signal, and simultaneous detection of multiple biotoxins in food based on single-cell are proposed for future research.

Zearalenone (ZEN) is one of the most common mycotoxin contaminants in food. For food safety, an efficient and environmental-friendly approach to ZEN degradation is significant. In this study, an Aspergillus niger strain, FS10, was stimulated with 1.0 μg/mL ZEN for 24 h, repeating 5 times to obtain a stressed strain, Zearalenone-Stressed-FS10 (ZEN-S-FS10), with high degradation efficiency. The results show that the degradation rate of ZEN-S-FS10 to ZEN can be stabilized above 95%. Through metabolomics analysis of the metabolome difference of FS10 before and after ZEN stimulation, it was found that the change of metabolic profile may be the main reason for the increase in the degradation rate of ZEN. The optimization results of degradation conditions of ZEN-S-FS10 show that the degradation efficiency is the highest with a concentration of 104 CFU/mL and a period of 28 h. Finally, we analyzed the degradation products by UPLC-q-TOF, which shows that ZEN was degraded into two low-toxicity products: C18H22O8S (Zearalenone 4-sulfate) and C18H22O5 ((E)-Zearalenone). This provides a wide range of possibilities for the industrial application of this strain.