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1.
Mikrochim Acta ; 191(8): 464, 2024 07 15.
Article in English | MEDLINE | ID: mdl-39007936

ABSTRACT

Rapid and high-sensitive Salmonella detection in milk is important for preventing foodborne disease eruption. To overcome the influence of the complex ingredients in milk on the sensitive detection of Salmonella, a dual-signal reporter red fluorescence nanosphere (RNs)-Pt was designed by combining RNs and Pt nanoparticles. After being equipped with antibodies, the immune RNs-Pt (IRNs-Pt) provide an ultra-strong fluorescence signal when excited by UV light. With the assistance of the H2O2/TMB system, a visible color change appeared that was attributed to the strong peroxidase-like catalytic activity derived from Pt nanoparticles. The IRNs-Pt in conjunction with immune magnetic beads can realize that Salmonella typhimurium (S. typhi) was captured, labeled, and separated effectively from untreated reduced-fat pure milk samples. Under the optimal experimental conditions, with the assay, as low as 50 CFU S. typhi can be converted to detectable fluorescence and absorbance signals within 2 h, suggesting the feasibility of practical application of the assay. Meanwhile, dual-signal modes of quantitative detection were realized. For fluorescence signal detection (emission at 615 nm), the linear correlation between signal intensity and the concentration of S. typhi was Y = 83C-3321 (R2 = 0.9941), ranging from 103 to 105 CFU/mL, while for colorimetric detection (absorbamce at 450 nm), the relationship between signal intensity and the concentration of S. typhi was Y = 2.9logC-10.2 (R2 = 0.9875), ranging from 5 × 103 to 105 CFU/mL. For suspect food contamination by foodborne pathogens, this dual-mode signal readout assay is promising for achieving the aim of convenient preliminary screening and accurate quantification simultaneously.


Subject(s)
Colorimetry , Milk , Salmonella typhimurium , Milk/microbiology , Milk/chemistry , Salmonella typhimurium/isolation & purification , Colorimetry/methods , Animals , Metal Nanoparticles/chemistry , Limit of Detection , Platinum/chemistry , Hydrogen Peroxide/chemistry , Fluorescence , Nanospheres/chemistry , Food Microbiology/methods , Food Contamination/analysis , Spectrometry, Fluorescence/methods
2.
ACS Nano ; 18(27): 17509-17520, 2024 Jul 09.
Article in English | MEDLINE | ID: mdl-38918939

ABSTRACT

There is growing concern about the distribution of nanoplastics (NPs) in the central nervous system (CNS), whereas intrusion is poorly understood. In this study, fluorescent-labeled polystyrene NPs (PS-NPs) were microinjected into different areas of zebrafish embryo to mimic different routes of exposure. PS-NPs were observed in the brain, eyes, and spinal cord through gametal exposure. It indicated that maternally derived PS-NPs were specially distributed in the CNS of zebrafish during early development. Importantly, these NPs were stranded in the CNS but not transferred to other organs during development. Furthermore, using neuron GFP-labeled transgenic zebrafish, colocalization between NPs and the neuron cells revealed that NPs were mostly enriched in the CNS surrounded but not the neurons. Even so, the intrusion of NPs into the CNS induced the significant upregulation of some neurotransmitter receptors, leading to an inhibited effect on the movement of zebrafish larvae. This work provides insights into understanding the intrusion and distribution of NPs in the CNS and the subsequent potential adverse effects.


Subject(s)
Central Nervous System , Polystyrenes , Zebrafish , Animals , Zebrafish/embryology , Central Nervous System/metabolism , Central Nervous System/drug effects , Polystyrenes/chemistry , Nanoparticles/chemistry , Nanoparticles/metabolism , Animals, Genetically Modified , Microplastics/toxicity
3.
Biotechnol J ; 19(1): e2300090, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37897200

ABSTRACT

Glycosylation is a ubiquitous posttranslational modification and plays an important role in many processes, such as protein stability, folding, processing, and trafficking. Among glycosylation types, O-glycosylation is difficult to analyze due to the complex glycan composition, low abundance and lack of glycosidases to remove the O-glycans. Many methods have been applied to analyze the O-glycosylation of membrane glycoproteins and secreted glycoproteins since the synthesis of O-glycosylation occurred in the Golgi apparatus. In recent years, some O-glycosylation has been reported in the nucleus. In this work, we present a proximity labeling strategy based on TurboID by combining core 1 ß1-3 galactosyltransferase (C1GalT1), which has been reported in the nucleus, to characterize nucleocytoplasmic O-glycosylation in living HeLa cells. The O-glycosylated protein C1GalT1 was biotinylated by the proximity labeling method in living HeLa cells overexpressing C1GalT1 fused by TurboID and enriched by streptavidin-coated beads. Following digestion with trypsin and mass spectrometry analysis, 68 high-confidence and 298 putative O-glycosylated sites were identified on 366 peptides mapped to 267 proteins. These results indicated that the proximity labeling method is a highly efficient technique to identify O-glycosylation. Furthermore, the finding of abundant O-glycosylation from nucleocytoplasmic proteins indicates a new pathway of O-glycosylation synthesis in cells.


Subject(s)
Glycoproteins , Protein Processing, Post-Translational , Humans , Glycosylation , HeLa Cells , Glycoproteins/chemistry , Mass Spectrometry
4.
Ecotoxicol Environ Saf ; 249: 114374, 2023 Jan 01.
Article in English | MEDLINE | ID: mdl-36508824

ABSTRACT

The Ya-Er Lake is a seriously polychlorinated dibenzo-p-dioxins/dibenzo-furans (PCDD/Fs) and mercury (Hg)-contaminated lake by pesticide and chlor-alkali plants in China. The oxidation pond method has been conducted to control pollution, moreover, the contaminated sediment was dredged and stacked, becoming a sediment stack yard for vegetable cultivation. To assess effects of oxidation pond method and dredging programme on pollution management and long-term risks of PCDD/Fs and Hg, the concentrations of PCDD/Fs, total Hg (THg), and methylmercury (MeHg) in soil and vegetable sampled from the sediment stack yard were measured and analyzed. Significantly positive relationships between concentrations of PCDD/Fs (p < 0.01), THg, and MeHg (p < 0.05) in edible parts of vegetable and soil were found, suggesting that bioaccumulation from contaminated soil derived from sediment dredging is important sources of PCDD/Fs and Hg in vegetable. Much higher PCDD/Fs (12 ± 9 pg/g dw) and Hg (THg, 0.14 ± 0.23 µg/g dw; MeHg,12.63 ± 13.31 ng/g dw) levels in vegetable were found compared with those from other contaminated regions, indicative of serious PCDD/Fs and Hg pollution in vegetable harvested from contaminated soil. Finally, the calculated provisional tolerable daily intake (PTDI) values showed higher health risk of PCDD/Fs and Hg exposure to local residents through consumption of purple and white flowering stalk, and oilseed rape. Our study established a good model to evaluate the long-term risks of PCDD/Fs and Hg. Moreover, the results indicate that the oxidation pond method and dredging programme were not effective to remove PCDD/Fs and Hg in sediment, which shed new light on management strategy of PCDD/Fs and Hg pollution in contaminated regions.


Subject(s)
Mercury , Polychlorinated Dibenzodioxins , Polychlorinated Dibenzodioxins/analysis , Dibenzofurans , Vegetables , Mercury/analysis , Geologic Sediments , China , Dibenzofurans, Polychlorinated , Soil
5.
Sci Total Environ ; 857(Pt 1): 159259, 2023 Jan 20.
Article in English | MEDLINE | ID: mdl-36220475

ABSTRACT

Bisphenol A (BPA) can disturb the estrogen receptor α (ERα)-mediated signaling pathway, which results in endocrine-disrupting effects and reproductive toxicity. Most BPA analogues as alternatives were evidenced to generate estrogenic activity as agonists or partial agonists of ERα. Recent studies indicated that certain BPA analogues, such as bisphenol M (BPM), bisphenol P (BPP), and bisphenol FL (BPFL), exhibited strong anti-estrogenic effects comparable with the typical antagonist 4-hydroxytamoxifen. However, conflicting findings were also observed for the compounds in different in vitro assays, and whether these BPA analogues can elicit an in vivo effect on ERα at environmentally relevant concentrations remains unknown. The underlying structural basis of estrogenic/anti-estrogenic activity should be further elucidated at the atomic level. To address these issues, we combined zebrafish-based in vivo and in silico methods to assess the effects of the compounds on ERα. The results show that the expressions of ERα-mediated downstream related genes in zebrafish embryos decreased after exposed to the compounds. Further molecular dynamics simulations were used to probe the antagonistic mechanisms of the compounds on ERα. The key H-bonding interactions were identified as important ligand recognition by ERα in the analysis of binding modes and binding free energy calculations. In summary, the current study provides preliminary in vivo evidence of fish species for the anti-estrogenic activity of certain BPA analogues.


Subject(s)
Estrogen Receptor alpha , Zebrafish , Animals , Estrogen Receptor alpha/metabolism , Zebrafish/metabolism , Benzhydryl Compounds/metabolism , Phenols/metabolism , Estrone
6.
Chemosphere ; 298: 134176, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35278457

ABSTRACT

Photo-induced holes (h+) oxidation is an efficient approach for perfluorooctanoic acid (PFOA; C7F15COOH) removal. To maintain a high amount of h+ on the surface of photocatalysts participating in the PFOA photodegradation could be a critical issue. Herein, a highly efficient spherical BiOBr-modified nano-TiO2 (P25) was synthesised and used for PFOA photodegradation through direct oxidation with h+. A high number of h+ could be generated and remain on the surface of P25/BiOBr due to the appropriate position of the conduction band (CB) and valence band (VB) levels between P25 and BiOBr. Meanwhile, PFOA molecules were coordinated to the P25/BiOBr's surface via unidentate binding, being directly activated and oxidised by h+, resulting in a decomposition yield of 99.5% (100 mg/L) under simulated solar light irradiation within 100 min, at the initial pH condition (3.5). A stepwise photodegradation pathway was proposed due to the significant intermediates detected as the short-chain perfluorinated carboxylic acids (C2-C7). Reactive oxygen species (ROS) generation, scavenging and trapping analysis indicated that the direct oxidation on h+ followed PFOA degradation. In a real aqueous environment of Tangxun lake (adjusted pH 3.5), stable common anions and natural organic matter (NOM) would restrain the PFOA photodegradation. However, adding 10 mg/L of NO3- or HA could reduce the inhibition effect of PFOA photodegradation. These findings gave an alternative strategy to drive an h+ directly oxidation to treat PFOA contaminated water bodies.


Subject(s)
Fluorocarbons , Bismuth , Fluorocarbons/chemistry , Photolysis , Titanium/chemistry
7.
J Steroid Biochem Mol Biol ; 213: 105966, 2021 10.
Article in English | MEDLINE | ID: mdl-34416373

ABSTRACT

The human estrogen receptor alpha (ERα) is an important regulator in breast cancer development and progression. The frequent ERα mutations in the ligand-binding domain (LBD) can increase the resistance of antiestrogen drugs, highlighting the need to develop new drugs to target ERα-positive breast cancer. In this study, we combined molecular docking, molecular dynamics simulations and binding free energy calculations to develop a structure-based virtual screening workflow to identify hit compounds capable of interfering with the recognition of ERα by the specific response element of DNA. A druggable pocket on the DNA binding domain (DBD) of ERα was identified as the potential binding site. The hits binding modes were further analyzed to reveal the structural characteristics of the DBD-inhibitor complexes. The core structure of the lead molecules was synthesized and was found to inhibit the E2-induced cell proliferation in MCF-7 cell lines. These findings provide an insight into the structural basis of ligand-ERα for alternate sites beyond the LBD-based pocket. The core structure proposed in this study could potentially be used as the lead molecule for further rational optimization of the antiestrogen drug structure with stronger binding of DBD and higher activity.


Subject(s)
Antineoplastic Agents/pharmacology , DNA/chemistry , Estrogen Antagonists/pharmacology , Estrogen Receptor alpha/chemistry , Neoplasm Proteins/chemistry , Response Elements , Antineoplastic Agents/chemical synthesis , Antineoplastic Agents/metabolism , Binding Sites , Cell Proliferation/drug effects , DNA/metabolism , Dose-Response Relationship, Drug , Estrogen Antagonists/chemical synthesis , Estrogen Antagonists/metabolism , Estrogen Receptor alpha/antagonists & inhibitors , Estrogen Receptor alpha/genetics , Estrogen Receptor alpha/metabolism , Female , Humans , Ligands , MCF-7 Cells , Molecular Docking Simulation , Molecular Dynamics Simulation , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/genetics , Neoplasm Proteins/metabolism , Nucleic Acid Conformation , Protein Binding , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Interaction Domains and Motifs , Structure-Activity Relationship , Thermodynamics , User-Computer Interface
8.
Ecotoxicol Environ Saf ; 171: 647-656, 2019 Apr 30.
Article in English | MEDLINE | ID: mdl-30658300

ABSTRACT

Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) have been reported to cause adverse health effects on wildlife as well as humans. Numerous studies demonstrated that PFOA and PFOS could interfere with the transcriptional activation of estrogen receptor α (ERα) by mimicking the function of endogenous ligand, whereas some reports suggested that the two compounds present non-estrogenic activities. These conflicting results bring a confusion to understand their molecular mechanism on the ERα-mediated signaling pathway. To address this issue, we performed the molecular docking and molecular dynamics simulations to elaborate the structural characteristics for the binding of PFOA and PFOS to ERα. Our results indicated that the two opposite binding orientations were modulated by the protonation states of key residue His524. In sub-acidic condition, PFOA and PFOS prefer to form the H-bonding interactions with the protonated His524, whereas Arg394 provided the H-bonding interactions for stable binding in sub-alkaline condition. Conformational analyses implied that the diverse binding modes were closely related to the conformational propensity of ERα for subsequent coactivator recruitment and transcription activation. Generally, our findings provide a flexible strategy to assess the pH impacts of microenvironment on the toxicities of perfluoroalkyl acids by their interactions with proteins.


Subject(s)
Alkanesulfonic Acids/metabolism , Caprylates/metabolism , Computer Simulation , Estrogen Receptor alpha/metabolism , Fluorocarbons/metabolism , Humans , Molecular Docking Simulation , Protein Conformation , Signal Transduction
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