Paeoniflorin recued hepatotoxicity under zinc oxide nanoparticles exposure via regulation on gut-liver axis and reversal of pyroptosis.

The risks of Zinc oxide nanoparticles (ZnO NPs) applications in biological medicine, food processing industry, agricultural production and the biotoxicity brought by environmental invasion of ZnO NPs both gradually troubled the public due to the lack of research on detoxification strategies. TFEB-regulated autophagy-pyroptosis pathways were found as the crux of the hepatotoxicity induced by ZnO NPs in our latest study. Here, our study served as a connecting link between preceding toxic target and the following protection mechanism of Paeoniflorin (PF). According to a combined analysis of network pharmacology/molecular docking-intestinal microbiota-metabolomics first developed in our study, PF alleviated the hepatotoxicity of ZnO NPs from multiple aspects. The hepatic inflammatory injury and hepatocyte pyroptosis in mice liver exposed to ZnO NPs was significantly inhibited by PF. And the intestinal microbiota disorder and liver metabolic disturbance were rescued. The targets predicted by bioinformatics and the signal trend in subacute toxicological model exhibited the protectiveness of PF related to the SIRT1-mTOR-TFEB pathway. These evidences clarified multiple protective mechanisms of PF which provided a novel detoxification approach against ZnO NPs, and further provided a strategy for the medicinal value development of PF.

Research of segmentation recognition of small disease spots on apple leaves based on hybrid loss function and CBAM.

Identification technology of apple diseases is of great significance in improving production efficiency and quality. This paper has used apple Alternaria blotch and brown spot disease leaves as the research object and proposes a disease spot segmentation and disease identification method based on DFL-UNet+CBAM to address the problems of low recognition accuracy and poor performance of small spot segmentation in apple leaf disease recognition. The goal of this paper is to accurately prevent and control apple diseases, avoid fruit quality degradation and yield reduction, and reduce the resulting economic losses. DFL-UNet+CBAM model has employed a hybrid loss function of Dice Loss and Focal Loss as the loss function and added CBAM attention mechanism to both effective feature layers extracted by the backbone network and the results of the first upsampling, enhancing the model to rescale the inter-feature weighting relationships, enhance the channel features of leaf disease spots and suppressing the channel features of healthy parts of the leaf, and improving the network's ability to extract disease features while also increasing model robustness. In general, after training, the average loss rate of the improved model decreases from 0.063 to 0.008 under the premise of ensuring the accuracy of image segmentation. The smaller the loss value is, the better the model is. In the lesion segmentation and disease identification test, MIoU was 91.07%, MPA was 95.58%, F1 Score was 95.16%, MIoU index increased by 1.96%, predicted disease area and actual disease area overlap increased, MPA increased by 1.06%, predicted category correctness increased, F1 Score increased by 1.14%, the number of correctly identified lesion pixels increased, and the segmentation result was more accurate. Specifically, compared to the original U-Net model, the segmentation of Alternaria blotch disease, the MIoU value increased by 4.41%, the MPA value increased by 4.13%, the Precision increased by 1.49%, the Recall increased by 4.13%, and the F1 Score increased by 2.81%; in the segmentation of brown spots, MIoU values increased by 1.18%, MPA values by 0.6%, Precision by 0.78%, Recall by 0.6%, and F1 Score by 0.69%. The spot diameter of the Alternaria blotch disease is 0.2-0.3cm in the early stage, 0.5-0.6cm in the middle and late stages, and the spot diameter of the brown spot disease is 0.3-3cm. Obviously, brown spot spots are larger than Alternaria blotch spots. The segmentation performance of smaller disease spots has increased more noticeably, according to the quantitative analysis results, proving that the model's capacity to segment smaller disease spots has greatly improved. The findings demonstrate that for the detection of apple leaf diseases, the method suggested in this research has a greater recognition accuracy and better segmentation performance. The model in this paper can obtain more sophisticated semantic information in comparison to the traditional U-Net, further enhance the recognition accuracy and segmentation performance of apple leaf spots, and address the issues of low accuracy and low efficiency of conventional disease recognition methods as well as the challenging convergence of conventional deep convolutional networks.

Spatial distribution and comparative analysis of Aconitum alkaloids in Fuzi using DESI-MSI and UHPLC-QTOF-MS.

Aconitum L. poisoning is a major type of poisoning caused by herbal medicines in many countries. However, despite its toxicity, Aconitum L. is still used because of its therapeutic value. Fuzi, the lateral root of Aconitum L., is one of the most important pharmacological parts. It is necessary for rational medication to figure out the types and contents of toxic Aconitum alkaloids (AAs) in Fuzi and its processed products. The present study aims to investigate the spatial distribution of toxic AAs in Fuzi and the quantification of AAs in various processing products through mass spectrometry methods. In this study, desorption electrospray ionization mass spectrometry imaging (DESI-MSI) was used to directly image the sections of raw Fuzi. The results showed a high content of diester alkaloids (DAs) and a relatively uniform distribution in the sections, while the content of monoester alkaloids (MAs) was low and uneven in the sections, distributed in the cortex, epidermis, vascular column, and other parts of the tissues. The content of non-ester alkaloids (NAs) was relatively minimum, and most of the NAs were distributed in the vascular column and the tightly connected cortex of the tissue. To further investigate the difference between raw and processed Fuzi, 60 known compounds were identified using UHPLC-QTOF-MS. The total contents of alkaloids in 7 processed Fuzi were lower than that in Shengfupian (SFP). Paofupian (PFP), Paotianxiong (PTX), Paofupian (PFP*), Danfupian (DFP), and Shufupian (SFP*) were the least similar. Zhengfupian (ZFP) and Chaofupian (CFP) had significantly reduced toxicity and increased efficacy compared with other processed products because the contents of active alkaloids in other processed products were also reduced. Understanding the distribution of metabolites and the composition changes after processing can guide users and herbal manufacturers to carefully choose the relatively safe and better therapeutic species of Fuzi. The information gathered from this study can contribute towards the improved and effective management of therapeutically important, nonetheless toxic, drugs such as Aconitum L.

Targeted preparation and recognition mechanism of broad-spectrum antibody specific to Aconitum alkaloids based on molecular modeling and its application in immunoassay.

Ester-type Aconitum alkaloids (AAs), the main medicinal ingredients of Aconitum L. herbs, could cause brain and heart damage in humans and animals and have raised concerns worldwide. In the present study, we aimed to produce a high-performance and broad-spectrum antibody and establish an immunoassay method of ester-type AAs, 3-succinyl aconitine (ACO-HS) was selected as an optimal hapten from five designed haptens comparing the similarity of stereo structure, electronic distribution, and physicochemical properties using the computer-aided molecular modeling technology. The monoclonal antibody (mAb) 1A9 exhibited broad-spectrum recognition specificity of 15 ester-type AAs was obtained and had a high sensitivity with the binding affinity (half-maximum inhibition concentration, IC50) of 0.73-130.36 µg L-1. Through molecular docking, it was found that mAb 1A9 and ester-type AAs showed a semi-enveloped structure through hydrogen bonds and hydrophobicity interaction. The amino acid residues that responsible for recognition were ARG107, GLU55, PRO113, VAL36, and SER64, and the critical structures to be recognized of AAs were acetyl group, benzoyl group, and N-linked carbon chains. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) based on mAb 1A9 allowed a sensitive determination of 15 ester-type AAs with the limit of detection (LOD) of 0.21-43.72 µg L-1, and it was suitable for the analysis of ester-type AAs in various Aconitum L. samples. These results provided an effective strategy for the preparation of targeted broad-spectrum antibodies of small molecules and proposed an icELISA method available for rapid, sensitive, and high-throughput detection of toxic ester-type AAs in Aconitum L. herbs.

Recombinant Antibody-Based and Computer-Aided Comprehensive Analysis of Antibody's Equivalent Recognition Mechanism of Alternariol and Alternariol Monomethyl Ether.

Alternariol (AOH) and alternariol monomethyl ether (AME) are two main Alternaria mycotoxins that endanger human health. In this study, a single-chain antibody fragment (scFv) capable of equivalently and specifically recognizing AOH and AME was first expressed, and its equivalent recognition mechanism was discussed. According to molecular docking and dynamic simulation, the C9 site, which was always exposed outside the binding cavity, made the structural differences between AOH and AME negligible. Due to the high similarity of structures, AOH and AME interacted with almost the same amino acids on the scFv; thus, the same interaction mode and interaction force were produced. This was considered to be the most critical reason for the equivalent recognition. Thus, the exposure of common structures was considered a potential strategy to obtain the equivalent recognition antibodies, and C9 was considered the key site in the process of hapten modification. These results laid a theoretical foundation for further research on antibodies against Alternaria mycotoxins. It could promote the rapid detection of AOH and AME in food and provide a new idea for targeted preparation of antibodies that could recognize multiple hazards with similar structures.

Progress in Immunoassays of Toxic Alkaloids in Plant-Derived Medicines: A Review.

Plants are the cradle of the traditional medicine system, assuaging human or animal diseases, and promoting health for thousands of years. However, many plant-derived medicines contain toxic alkaloids of varying degrees of toxicity that pose a direct or indirect threat to human and animal health through accidental ingestion, misuse of plant materials, or through the food chain. Thus, rapid, easy, and sensitive methods are needed to effectively screen these toxic alkaloids to guarantee the safety of plant-derived medicines. Antibodies, due to their inherent specificity and high affinity, have been used as a variety of analytical tools and techniques. This review describes the antigen synthesis and antibody preparation of the common toxic alkaloids in plant-derived medicines and discusses the advances of antibody-based immunoassays in the screening and detection of toxic alkaloids in plants or other related matrices. Finally, the limitations and prospects of immunoassays for toxic alkaloids are discussed.

Development of a sensitive and rapid fluorescence polarization immunoassay for high throughput screening eight glucocorticoids in beef.

In this study, a sensitive, rapid, homogeneous, and high-throughput fluorescence polarization immunoassay (FPIA) for the rapid screening of eight glucocorticoids (GCs) in beef samples was successfully established. Two tracers including 5-aminofluorescein-labeled dexamethasone (5-AF-DMS) and fluorescein isothiocyanate-labeled dexamethasone (FITC-DMS) were studied to select appropriate antibody-tracer pairs using four previously produced broad-specific monoclonal antibodies. An optimal combination of the antibody 12D9 and the tracer FITC-DMS was selected. Under optimal detection conditions, the half inhibitory concentrations of dexamethasone (DMS), betamethasone (BMS), prednisolone (PNS), hydrocortisone (HCS), beclomethasone (BCMS), cortisone (CS), 6-α-methylprednisone (6-α-MPNS), fludrocortisone acetate (HFCS) were 1.00, 2.17, 3.49, 12.45, 1.20, 5.66, 6.85 and 3.45 ng/mL, respectively. The average recoveries of the proposed method in beef samples ranged from 77.3-91.7% with the coefficient of variation less than 12%. The developed FPIA was time-saving that could be completed within 10 min. The FPIA was applied to beef samples and showed a good correlation with liquid chromatography-tandem mass spectrometry (LC-MS/MS) (R2 = 0.9894). Thus, the proposed method provides a rapid, reliable, sensitive, and high-throughput screening tool for the simultaneous screening of eight GCs in beef, which shows great potential in the food safety analysis.

A comprehensive review on acquisition of phenotypic information of Prunoideae fruits: Image technology.

Fruit phenotypic information reflects all the physical, physiological, biochemical characteristics and traits of fruit. Accurate access to phenotypic information is very necessary and meaningful for post-harvest storage, sales and deep processing. The methods of obtaining phenotypic information include traditional manual measurement and damage detection, which are inefficient and destructive. In the field of fruit phenotype research, image technology is increasingly mature, which greatly improves the efficiency of fruit phenotype information acquisition. This review paper mainly reviews the research on phenotypic information of Prunoideae fruit based on three imaging techniques (RGB imaging, hyperspectral imaging, multispectral imaging). Firstly, the classification was carried out according to the image type. On this basis, the review and summary of previous studies were completed from the perspectives of fruit maturity detection, fruit quality classification and fruit disease damage identification. Analysis of the advantages and disadvantages of various types of images in the study, and try to give the next research direction for improvement.

Electrical impedance spectroscopy (EIS) in plant roots research: a review.

Nondestructive testing of plant roots is a hot topic in recent years. The traditional measurement process is time-consuming and laborious, and it is impossible to analyze the state of plant roots without destroying the sample. Recent studies have shown that as an excellent nondestructive measurement method, although electrical impedance spectroscopy (EIS) has made great achievements in many botanical research fields such as plant morphology and stress resistance, there are still limitations. This review summarizes the application of EIS in plant root measurement. The experiment scheme, instrument and electrode, excitation frequency range, root electrical characteristics, equivalent circuit, and combination of EIS and artificial intelligence (AI) are discussed. Furthermore, the review suggests that future research should focus on miniaturization of measurement equipment, standardization of planting environment and intelligentization of root diagnosis, so as to better apply EIS technology to in situ root nondestructive measurement.

A highly specific and sensitive indirect competitive monoclonal ELISA for the detection of brombuterol in animal feed, swine urine, pork and liver samples.

Brombuterol, a new ß-adrenergic agonist to enhance animal growth and increase feeding efficiency, is forbidden as an additive in animal feed for livestock production due to its adverse effects on consumers. In this study, a highly specific and sensitive monoclonal antibody was obtained and an indirect competitive monoclonal ELISA was developed to detect brombuterol, with an IC50 value of 0.1 µg/kg (µg/L) and no cross-reactivity to other structurally related ß-adrenergic agonists. The average recovery of brombuterol using the icELISA method ranged from 72.9% to 106.4% with the coefficient of variation lower than 18.9%, which was determined by analysing spiked animal feed, swine urine, pork and liver samples (n = 5). Finally, the icELISA gave results having a good correlation with those obtained by liquid chromatography-tandem mass spectrometry. These results demonstrated that the developed icELISA for the detection of brombuterol is highly specific, sensitive, and reliable, indicating good potential for use in the area of food safety to improve consumer protection.

Fluorescent lateral flow immunoassay based on gold nanocluster for detection of pyrrolizidine alkaloids.

An ultrasensitive and rapid fluorescent immunoassay based on a broad-spectrum monoclonal antibody (mAb) was developed to detect pyrrolizidine alkaloids (PAs) in honey samples. First, Discovery Studio software was used to analyze and predict the target hapten, and retrorsine (RTS) was selected to react with succinic anhydride (HS) for hapten synthesization. A sensitive and broad-spectrum monoclonal antibody (mAb 13E1) was obtained for nine PAs. Then, fluorescent gold nanoclusters (AuNCs) were conjugated with mAb as a label probe and used in establishing a qualitative and quantitative lateral flow immunoassay (AuNCs-LFIA) for the determination of four PAs (retrorsine, platyphylline, senecionine, integerrimine) in honey within 14 min. The limits of detection (LOD) were 0.083 µg/kg. The recovery in spiked honey samples were 87.98-119.57%, with coefficients of variation of ≤ 11.5%. A total of 45 commercial import honey samples from nine different countries were tested through AuNCs-LFIA and UPLC-MS/MS method, and satisfactory consistency (R2 = 0.995) was obtained. The rates of positive samples were 55.56% (25/45), and the average concentrations of four PAs were 3.24-46.47 µg/kg. This ultrasensitive multi-PA method provides an alternative analytical tool for evaluating the human risk posed by the consumption of PA-contaminated honey.

Toxic Prediction of Pyrrolizidine Alkaloids and Structure-Dependent Induction of Apoptosis in HepaRG Cells.

Pyrrolizidine alkaloids (PAs) are common phytotoxins and could cause liver genotoxicity/carcinogenicity following metabolic activation. However, the toxicity of different structures remains unclear due to the wide variety of PAs. In this study, the absorption, distribution, metabolism, excretion, and toxicity (ADMET) of 40 PAs were analyzed, and their toxicity was predicted by Komputer Assisted Technology (TOPKAT) using Discovery Studio software. The in silico results showed that all PAs except retronecine had good intestinal absorption, and all PAs were predicted to have different toxicity ranges. To verify the predictive results, 4 PAs were selected to investigate cell injury and possible mechanisms of the differentiation in HepaRG cells, including retronecine type of twelve-membered cyclic diester (retrorsine), eleven-membered cyclic diester (monocrotaline), noncyclic diester (retronecine), and platynecine type (platyphylline). After 24 h exposure, retronecine-type PAs exhibited concentration-dependent cytotoxicity. The high-content screening assay showed that cell oxidative stress, mitochondrial damage, endoplasmic reticulum stress, and the concentration of calcium ions increased, and neutral lipid metabolism was changed notably in HepaRG cells. Induced apoptosis by PAs was indicated by cell cycle arrest in the G2/M phase, disrupting the mitochondrial membrane potential. Overall, our study revealed structure-dependent cytotoxicity and apoptosis after PA exposure, suggesting that the prediction results of in silico have certain reference values for compound toxicity. A 1,2-membered cyclic diester seems to be a more potent apoptosis inducer than other PAs.

Indirect competitive enzyme-linked immunosorbent assay based on a broad-spectrum monoclonal antibody for tropane alkaloids detection in pig urine, pork and cereal flours.

In this study, an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a broad-spectrum monoclonal antibody for tropane alkaloids (TAs) was established for the rapid screening of atropine, scopolamine, homatropine, apoatropine, anisodamine, anisodine and L-hyoscyamine residues in pig urine, pork and cereal flour samples through a simple sample preparation procedure. The half inhibitory concentrations of atropine, homatropine, L-hyoscyamine, apoatropine, scopolamine, anisodamine and anisodine were 0.05, 0.07, 0.14, 0.14, 0.24, 5.30 and 10.15 ng mL-1, respectivelyThe detection and quantitative limits of this method for TAs in samples were 0.18-73.18 and 0.44-74.77 µg kg-1. The spiked recoveries ranged from 69.88% to 147.93%, and the coefficient of variations were less than 14%. Good correlation (R2 = 0.9929) between the results of the ic-ELISA and the high performance liquid chromatography-tandem mass spectrometry support the reliability of the developed ic-ELISA method.

Phenotypic techniques and applications in fruit trees: a review.

Phenotypic information is of great significance for irrigation management, disease prevention and yield improvement. Interest in the evaluation of phenotypes has grown with the goal of enhancing the quality of fruit trees. Traditional techniques for monitoring fruit tree phenotypes are destructive and time-consuming. The development of advanced technology is the key to rapid and non-destructive detection. This review describes several techniques applied to fruit tree phenotypic research in the field, including visible and near-infrared (VIS-NIR) spectroscopy, digital photography, multispectral and hyperspectral imaging, thermal imaging, and light detection and ranging (LiDAR). The applications of these technologies are summarized in terms of architecture parameters, pigment and nutrient contents, water stress, biochemical parameters of fruits and disease detection. These techniques have been shown to play important roles in fruit tree phenotypic research.

The Biosynthesis Pathway of Swainsonine, a New Anticancer Drug from Three Endophytic Fungi.

Swainsonine (SW) is the principal toxic ingredient of locoweed plants that causes locoism characterized by a disorder of the nervous system. It has also received widespread attention in the medical field for its beneficial anticancer and antitumor activities. Endophytic fungi, Alternaria sect. Undifilum oxytropis isolated from locoweeds, the plant pathogen Slafractonia leguminicola, and the insect pathogen Metarhizium anisopliae, produce swainsonine. Acquired SW by biofermentation has a certain foreground and research value. This paper mainly summarizes the local and foreign literature published thus far on the swainsonine biosynthesis pathway, and speculates on the possible regulatory enzymes involved in the synthesis pathway within these three fungi in order to provide a new reference for research on swainsonine biosynthesis by endophytic fungi.

The Genome of Undifilum oxytropis Provides Insights into Swainsonine Biosynthesis and Locoism.

Undifilum oxytropis is a fungal endophyte of locoweeds. It produces swainsonine, which is the principal toxic ingredient of locoweeds. However, the genes, pathways and mechanisms of swainsonine biosynthesis are not known. In this study, the genome of U. oxytropis was firstly sequenced and assembled into a 70.05 megabases (Mb) draft genome, which encoded 11,057 protein-coding genes, and 54% of them were similar to current publicly available sequences. U. oxytropis genes were annotated and 164 putative genes were annotated into enzymes, such as Saccharopine dehydrogenase, Saccharopine oxidase, and Pyrroline-5-carboxylate reductase, hypothesized to be involved in the biosynthesis pathway of swainsonine. The genome sequence and gene annotation of U. oxytropis will provide new insights into functional analyses. The characterization of genes in swainsonine biosynthesis will greatly facilitate locoweed poisoning research and help direct locoism management.

Hypoxia­inducible adrenomedullin ameliorates the epithelial-to-mesenchymal transition in human proximal tubular epithelial cells.

Renal tubular epithelial cells can enter the epithelial­to­mesenchymal transition (EMT) in response to chronic hypoxia. EMT is a process which involves the phenotypic conversion of epithelial cells, that is believed to have an important role in renal fibrosis. However, the underlying mechanisms of the involvement of EMT in renal fibrosis remain to be elucidated. Adrenomedullin (AMD) has been implicated in renal fibrosis and is induced by hypoxia. The aims of the present study were to determine whether ADM signaling was active in human proximal tubular epithelial cells cultured under hypoxic conditions, and to observe the activity of ADM during EMT. The expression levels of ADM were significantly increased, in a time­dependent manner, in HK­2 and HKC human proximal tubular epithelial cells, cultured under hypoxic conditions. Overexpression of exogenous ADM was accompanied by increased expression levels of the epithelial markers E­cadherin and tight junction protein­1, and decreased expression levels of the mesenchymal markers vimentin and α­smooth muscle actin, during hypoxia. Knock­down of ADM expression by small hairpin RNA, or co­administration of an ADM peptide inhibitor, in HK­2 cells significantly exacerbated hypoxia­induced EMT, as compared to the lack of effect observed in the untransfected controls. ADM was shown to suppress EMT by inhibiting the activation of extracellular signal­regulated kinase (ERK), and this effect was prevented by the ERK activator apigenin. The results of the present study suggest that ADM has an important role in promoting EMT in hypoxic human proximal tubular epithelial cells. ADM may therefore represent a novel therapeutic target in the treatment of injured kidneys.