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The fungicide phenamacril has been employed to manage Fusarium and mycotoxins in crops, leading to persistent residues in the environment and plants. Detecting phenamacril is pivotal for ensuring environmental and food safety. In this study, haptens and artificial antigens were synthesized to produce antiphenamacril monoclonal antibodies (mAbs). Additionally, gold nanoparticles coated with a polydopamine shell were synthesized and conjugated with mAbs, inducing fluorescence quenching in quantum dots. Moreover, a dual-readout immunochromatographic assay that combines the positive signal from fluorescence with the negative signal from colorimetry was developed to enable sensitive and precise detection of phenamacril within 10 min, achieving detection limits of 5 ng/mL. The method's reliability was affirmed by using spiked wheat flour samples, achieving a limit of quantitation of 0.05 mg/kg. This analytical platform demonstrates high sensitivity, outstanding accuracy, and robust tolerance to matrix effects, making it suitable for the rapid, onsite, quantitative screening of phenamacril residues.
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Colorimetria , Contaminação de Alimentos , Fungicidas Industriais , Resíduos de Praguicidas , Fungicidas Industriais/análise , Contaminação de Alimentos/análise , Colorimetria/métodos , Resíduos de Praguicidas/análise , Anticorpos Monoclonais/química , Cromatografia de Afinidade/métodos , Cromatografia de Afinidade/instrumentação , Fluorescência , Triticum/química , Nanopartículas Metálicas/química , Ouro/química , Limite de Detecção , Farinha/análiseRESUMO
BACKGROUND & AIMS: The association between antibiotic exposure and inflammatory bowel disease (IBD) remains controversial, especially whether there is a dose-response relationship. We aimed to conduct a systematic review and meta-analysis to thoroughly evaluate the risk of new-onset IBD associated with antibiotic exposure. METHODS: Four databases were searched from their inception to September 30, 2023 for all relevant studies. The risk estimates were pooled together using random-effects models, and pooled odds ratios (ORs) with 95% confidence intervals (CIs) were calculated, stratified by IBD subtype, age, exposure period, study type, and antibiotic classes. Dose-response relationship between the number of antibiotic prescriptions and IBD risk was assessed using generalized least squares regression analysis. RESULTS: Twenty-eight studies involving 153,027 patients with IBD were included. Antibiotic exposure was significantly associated with an increased risk of new-onset IBD for prescription-based studies (pooled OR, 1.41; 95% CI, 1.29-1.53) and for questionnaire-based studies (pooled OR, 1.35; 95% CI, 1.08-1.68). This association existed for both Crohn's disease and ulcerative colitis, as well as in children and adults for prescription-based studies. The majority of antibiotic classes were associated with an increased IBD risk, with metronidazole (OR, 1.70; 95% CI, 1.38-2.10) and quinolones (OR, 1.56; 95% CI, 1.37-1.77) having relatively higher risk estimates. A positive nonlinear dose-response association was observed between the number of antibiotic prescriptions and IBD risk. CONCLUSIONS: Antibiotic exposure was significantly associated with an increased risk of new-onset IBD, and a positive nonlinear dose-response relationship was observed. Antibiotic stewardship may be important for reducing IBD risk.
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Later flow immunochromatographic assay has been widely used in clinical, environmental, and other diagnostic applications owing to its high sensitivity and throughput. However, most immunoassays operate in the "turn-off" mode for detecting targets of low molecular weight. The signal intensity decreases as the analyte concentration increases, which poses a challenge for achieving ultrasensitive detection at low concentrations and is counterintuitive to new users. In this work, a fluorometric immunochromatographic assay (FICA) is developed to simultaneously read "turn-on" fluorescent and "turn-off" colorimetric signals, where ZnCdSe/ZnS quantum dots act as fluorescence donors and gold nanoparticles (AuNPs) act as quenchers. The fluorescent signal (excitation/emission wavelengths of 365/525 nm) is positively correlated with analytes' concentration. Taking sibutramine (SBT) as the analysis target, the visual limit of detection for SBT reached 3.9 ng/mL, and the limit of Quantitation was 5.0 ng/mg in spiked samples. The developed FICA achieves a high sensitivity in SBT detection, which is much lower than that of the colloidal gold-based immunochromatographic assay. This dual-function detection mode has great potential to be used as a rapid on-site semiquantitative method, providing an alternative mode for the determination of low levels of target analytes.
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In this study, we develop a competitive ratiometric fluorescent lateral flow immunoassay (CRF-LFIA) based on dual emission fluorescence signal, which has great advantage in visual and quantitative detection of Chlorothalonil (CTN). Red-emitted fluorescent magnetic nanobeads (FMNBs) and green-emitted aggregation-induced emission fluorescent microsphere (AIEFM) are synthesized and conjugated to antibodies and antigens respectively, resulting in competitive binding with the analyte. The ratiometric fluorescence signal which comes from the overlap of these two fluorescence emissions. FMNBs probes also provide immunomagnetic separation (IMS) to enrich the analysts and resist complex matrix effects. This strip generates a visually discernible yellow-to-green fluorescence color change in the presence of CTN (2 ng/mL), which could be incisively observed by naked eye. Moreover, the limit of detection (LOD) reached 0.152 ng/mL by measurement of color (Red-Green-Blue, RGB) signals. Method validation shows a good correlation between CRF-LFIA and LC-MS/MS.
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Corantes Fluorescentes , Nitrilas , Espectrometria de Massas em Tandem , Cromatografia Líquida , Imunoensaio/métodos , Limite de DetecçãoRESUMO
Background/Aims: Irritable bowel syndrome (IBS) is accepted as a disorder of gut-brain interactions. Berberine and rifaximin are non-absorbed antibiotics and have been confirmed effective for IBS treatment, but there is still lack of direct comparison of their effects. This study aims to compare the effect of the 2 drugs on the alteration of gut-brain axis caused by gut microbiota from IBS patients. Methods: Germ-free rats received fecal microbiota transplantation from screened IBS patients and healthy controls. After 14 days' colonization, rats were administrated orally with berberine, rifaximin or vehicle respectively for the next 14 days. The visceral sensitivity was evaluated, fecal microbiota profiled and microbial short chain fatty acids were determined. Immunofluorescence staining and morphological analysis were performed to evaluate microglial activation. Results: Visceral hypersensitivity induced by IBS-fecal microbiota transplantation was relieved by berberine and rifaximin, and berberine increased sucrose preference rate. Microbial α-diversity were reduced by both drugs. Compared with rifaximin, berberine significantly changed microbial structure and enriched Lachnoclostridium. Furthermore, berberine but not rifaximin significantly increased fecal concentrations of acetate and propionate acids. Berberine restored the morphological alterations of microglia induced by dysbiosis, which may be associated with its effect on the expression of microbial gene pathways involved in peptidoglycan biosynthesis. Rifaximin affected neither the numbers of activated microglial cells nor the microglial morphological alterations. Conclusions: Berberine enriched Lachnoclostridium, reduced the expression of peptidoglycan biosynthesis genes and increased acetate and propionate. The absence of these actions of rifaximin may explain the different effects of the drugs on microbiota-gut-brain axis.
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Gray mold infected with Botrytis cinerea frequently appears on fruits and vegetables throughout the supply chain after harvest, leading to economic losses. Biological control of postharvest disease with phytochemicals is a promising approach. CA (cinnamaldehyde) is a natural phytochemical with medicinal and antimicrobial activity. This study evaluated the effect of CA in controlling B. cinerea on fresh pepper fruit. CA inhibited B. cinerea growth in vitro significantly in a dose- (0.1-0.8 mM) and time-dependent (6-48 h) manner, with an EC50 (median effective concentration) of 0.5 mM. CA induced the collapse and breakdown of the mycelia. CA induced lipid peroxidation resulting from ROS (reactive oxygen species) accumulation in mycelia, further leading to cell leakage, evidenced by increased conductivity in mycelia. CA induced mycelial glycerol accumulation, resulting in osmotic stress possibly. CA inhibited sporulation and spore germination resulting from ROS accumulation and cell death observed in spores. Spraying CA at 0.5 mM induced a defense response in fresh pepper fruits, such as the accumulation of defense metabolites (flavonoid and total phenols) and an increase in the activity of defense enzymes (PAL, phenylalanine ammonia lyase; PPO, polyphenol oxidase; POD, peroxidase). As CA is a type of environmentally friendly compound, this study provides significant data on the activity of CA in the biocontrol of postharvest gray mold in peppers.
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A gold nanoparticle (AuNP) based immunochromatographic assay strip is a valuable tool for monitoring chemicals in foods. However, the sensitive ICA strip for SBT is rarely reported due to the fact that monoclonal antibodies (mAbs) against SBT with high affinity are commercially unavailable. Herein, a monoclonal antibody against SBT was prepared through a designed hapten with a carboxyl end-capped space arm. The obtained mAb showed high affinity for SBT and N-desmethylsibutramine, a metabolite of SBT. Furthermore, a series of core-shell NPs, polydopamine (PDA) coated AuNPs (PDA/AuNPs) with controlled shell thickness and packing density were synthesized. The obtained PDA/AuNP-mAb conjugate demonstrated high tolerance to salt and good stability in a wide pH range, which is beneficial for improving the matrix interference common in ICA. As a result, PDA/AuNP-based ICA could quantify SBT in the range of 3.39-69.60 ng mL-1 with a limit of detection (LOD) of 0.98 ng mL-1. This novel ICA improved the sensitivity of the traditional AuNP-based ICA by nearly 12 times. Method validation was conducted with spiked samples of slimming food and human serum and compared with HPLC-MS/MS. Consistent results indicated that high sensitivity, accuracy, and reliability of the PDA/AuNP-based ICA approach were achieved. To the best of our knowledge, this study reported the most sensitive immunoassay for SBT thus far.
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Ouro , Nanopartículas Metálicas , Humanos , Ouro/química , Reprodutibilidade dos Testes , Colorimetria , Espectrometria de Massas em Tandem , Nanopartículas Metálicas/química , Imunoensaio/métodos , Limite de Detecção , DietaRESUMO
E. sinensis, normally harvested in October and November, is an economic aquatic product in China. Pond culture has been widely applied for the production of E. sinensis, wherein a stable food supply for crabs is provided. In order to improve the nutritional quality of E. sinensis products, this study evaluated the effect of the local pond culture on the nutritive profiles of E. sinensis and screened out the best harvest time for the nutrient-rich crabs, thereby guiding the local crab industry to improve its aquaculture mode and harvest strategy. The results indicated that pond culture enhanced the levels of protein, amino acids, and specific organic acid derivatives, and reduced the levels of peptides and polyunsaturated fatty acids (PUFAs). Compared with E. sinensis harvested in October, peptide levels were significantly increased, whereas sugar, phenolic acid, and nucleotide levels were decreased in those harvested in November. Overall, the study revealed that the nutritive profile of the pond-reared E. sinensis was significantly modulated by a high-protein diet, thus lacking the diversity of metabolites. Additionally, October could be more appropriate for harvesting E. sinensis than November.
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Abnormal brain-gut interaction is considered the core pathological mechanism behind the disorders of gut-brain interaction (DGBI), in which the intestinal microbiota plays an important role. Microglia are the "sentinels" of the central nervous system (CNS), which participate in tissue damage caused by traumatic brain injury, resist central infection and participate in neurogenesis, and are involved in the occurrence of various neurological diseases. With in-depth research on DGBI, we could find an interaction between the intestinal microbiota and microglia and that they are jointly involved in the occurrence of DGBI, especially in individuals with comorbidities of mental disorders, such as irritable bowel syndrome (IBS). This bidirectional regulation of microbiota and microglia provides a new direction for the treatment of DGBI. In this review, we focus on the role and underlying mechanism of the interaction between gut microbiota and microglia in DGBI, especially IBS, and the corresponding clinical application prospects and highlight its potential to treat DGBI in individuals with psychiatric comorbidities.
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Encefalopatias , Microbioma Gastrointestinal , Síndrome do Intestino Irritável , Humanos , Síndrome do Intestino Irritável/terapia , Microglia , EncéfaloRESUMO
Traditional quantum dot-based lateral flow immunoassay (QD-LFIA) is limited to signal loss in part by the blinking, photobleaching and oxidative quenching of QD probes. Inspired by the good application of silver deposition on QD surfaces in tissue imaging, and in the context of improving the assay performance without compromising the simplicity and practicality, we report that introducing the QD-silver combination to the LFIA system, has the advantages of accuracy improvement, signal enhancement and user friendliness promotion, but maintains the cost-effective property and commercial accessibility of QD-LFIA. The effect was shown by using CdSe/ZnS QD-LFIA coupled with anti-sodium pentachlorophenate antibody, which provided a 4-fold improvement in the signal, a 2.5-fold improvement in the detection limit and a zero false-negative rate for sodium pentachlorophenate analysis in chicken samples. The proposed LFIA integrates the possibilities of colorimetric and fluorometric detection with different detection limits (fluorometric at 10 ng/mL and colorimetric at 4 ng/mL) and with acceptable detection times (fluorometric at 12 min and colorimetric at 27 min). The current results indicate that this QD-silver combined LFIA is complementary to conventional fluorescence LFIA and could be an inexpensive, versatile, and sensitive alternative.
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This study investigated the effects of berberine (BBR) on pancreatic cancer (PC) lung metastasis and explored the underlying mechanisms, using a BALB/C-nu/nu nude mouse model injected with PC cells (AsPC-1). Intragastric administration of BBR dose-dependently improves survival of mice intravenously injected with AsPC-1 cells, and reduces lung metastasis. Especially, BBR significantly reduces lung infiltration of circulating tumor cells (CTCs) 24 h after AsPC-1 cells injection. In vitro, tumor cells (TCs) trigger endothelial barrier disruption and promote trans-endothelial migration of CFSE-labeled TCs. BBR treatment effectively ameliorates TC-induced endothelial disruption, an effect that is diminished by inhibiting transforming growth factor-ß receptor 1 (TGFBR1). Blocking TGFBR1 blunts the anti-metastatic effect of BBR in vivo. Mechanistically, BBR binds to the intercellular portion of TGFBR1, suppresses its enzyme activities, and protects endothelial barrier disruption by TCs which express higher levels of TGF-ß1. Hence, BBR might be a promising drug for reducing PC lung metastasis in clinical practice.
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In this study, a designed hapten possessing the classic structure of PDE-5 inhibitors was synthesized. A monoclonal antibody (mAb) with broad recognition for six PDE-5 inhibitors was further produced. For the determination of lodenafil, methisosildenafil, mirodenafil, udenafil and tadalafil, the limit of detection (LOD) and IC50 ranged from 1.01 to 26.91 ng mL-1 and 12.75 to 278 ng mL-1, respectively. Thereafter, a quantum dot bead-based lateral flow immunoassay (QB-LFIA) was developed, which improved the LOD and IC50 to 0.32-6.52 ng mL-1 and 7.45-133.8 ng mL-1, respectively. Method validation was conducted using honey and capsule samples spiked with PDE-5 inhibitors, and the recoveries of the intra- and inter-assays ranged from 81.01% to 108.16%, with coefficients of variation below 12.71%. In addition, the validity and the consistency have been confirmed with a comparison between QB-LFIA and HPLC-MS/MS (R2 = 0.9957). Furthermore, the developed QB-LFIA was employed for the inspection of real products, and several samples were found to be adulterated with lodenafil and methisosildenafil.
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Inibidores da Fosfodiesterase 5 , Pontos Quânticos , Alimento Funcional , Imunoensaio/métodos , Inibidores da Fosfodiesterase 5/química , Pontos Quânticos/química , Espectrometria de Massas em TandemRESUMO
Polychlorinated biphenyls (PCBs) are persistent organic pollutants (POPs) with multiple variants, which may be harmful to human health by absorption and bioaccumulation. To ensure food safety, it is necessary to develop multi-residue immunoassays for broad recognition of PCBs. In this study, by mimicking the generic core structure of PCBs, three haptens have been designed and synthesized for monoclonal antibody (mAb) generation. A carboxylic acid derivative of PCB80 was a hapten that induced a mAb with broad recognition of PCBs. The results of ELISA further identified that the mAb could recognize 11 different kinds of PCBs; half-maximal inhibition concentrations (IC50) ranged from 33.12 to 476.42 ng/mL. Subsequently, using aggregation-induced emission luminogen (AIEgen) nanobeads as the tracer for the output signal, the IC50 value of the various PCBs was improved to 6.38-252.1 ng/mL. The limit of detection (LOD) varied from 0.32 to 42.15 ng/mL. Recoveries of 76.90-95.74% and intra-assay coefficients of variation of 8.5-14.4% were obtained with spiked chicken and crab meat samples. Matrix interference was eliminated by dilution, and no false-positive and false-negative results were observed. The developed assay provides a simple, broad-spectrum, and sensitive tool for detecting PCBs, with high-throughput possibilities for large-scale screening of PCBs in food.
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Braquiúros , Bifenilos Policlorados , Animais , Anticorpos Monoclonais , Galinhas , Haptenos , Imunoensaio/métodos , Bifenilos Policlorados/análise , Alimentos Marinhos/análiseRESUMO
To study the friction and wear performance of carbon fiber reinforced friction materials under different working conditions, paper-based friction materials with different fibers were prepared. Experiments on the SAE#2 test bench were conducted to study the infectors including friction torques, surface temperature, coefficient of friction (COF), and surface morphologies. The results were analyzed, which indicated that the carbon fiber reinforced friction material could provide a higher friction torque and a lower temperature rising rate under the applied high pressure and high rotating speed conditions. As the pressure increased from 1 MPa to 2.5 MPa, the friction torque of plant fiber reinforced material increased by 150%, the friction torque of carbon fiber reinforced material increased by 400%, and the maximum temperature of plant fiber reinforced and carbon fiber reinforced material reached the highest value at 1.5 MPa. Thus, carbon fibers not only improved the COF and friction torque performance but also had advantages in avoiding thermal failure. Meanwhile, carbon fiber reinforced friction materials can provide a more stable COF as its variable coefficient (α) only rose from 38.18 to 264.62, from 1 MPa to 2.5 MPa, which was much lower than the natural fiber reinforced friction materials. Simultaneously, due to the good dispersion and excellent mechanical properties of PAN chopped carbon fibers, fewer pores formed on the initial surface, which improved the high wear resistance, especially in the intermedia disc.
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Background: Irritable bowel syndrome (IBS) is a common gastrointestinal condition. Studies regarding the treatment of IBS with probiotics have not yielded consistent results, and the best probiotics has not yet been confirmed. Therefore, we performed a network meta-analysis (NMA) to assess the relative rank order of different probiotics for IBS. Method: We searched for RCTs on the efficacy of probiotics for IBS until August 25, 2021. The primary outcome was the symptom relief rate, as well as global symptoms, abdominal pain, bloating, and straining scores. The NMA was conducted using Stata 15.0. We also used meta-regression to explore whether the treatment length and dose influenced the efficacy. Results: Forty-three RCTs, with 5,531 IBS patients, were included in this analysis. Firstly, we compared the efficacy of different probiotic species. B.coagulans exhibited the highest probability to be the optimal probiotic specie in improving IBS symptom relief rate, as well as global symptom, abdominal pain, bloating, and straining scores. In regard to the secondary outcomes, L.plantarum ranked first in ameliorating the QOL of IBS patients, but without any significant differences compared with other probiotic species in standardized mean differences (SMD) estimates. Moreover, patients received L.acidophilus had lowest incidence of adverse events. The meta-regression revealed that no significant differences were found between participants using different doses of probiotics in all outcomes, while the treatment length, as a confounder, can significantly influence the efficacy of probiotics in ameliorating abdominal pain (Coef = -2.30; p = 0.035) and straining (Coef = -3.15; p = 0.020) in IBS patients. Thus, we performed the subgroup analysis on treatment length subsequently in these two outcomes, which showed that efficacy of B.coagulans using 8 weeks ranked first both in improving the abdominal pain and straining scores. Additionally, B. coagulans still had significant efficacy compared to different types of probiotic combinations in present study. Conclusions: The findings of this NMA suggested that B.coagulans had prominent efficacy in treating IBS patients, and incorporating B.coagulans into a probiotic combination, or genetically engineering it to amplify its biological function may be a future research target to treat IBS patients. With few direct comparisons available between individual therapies today, this NMA may have utility in forming treatment guideline for IBS with probiotics.
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Síndrome do Intestino Irritável , Probióticos , Dor Abdominal/etiologia , Dor Abdominal/terapia , Humanos , Síndrome do Intestino Irritável/diagnóstico , Síndrome do Intestino Irritável/tratamento farmacológico , Metanálise em Rede , Probióticos/uso terapêutico , Qualidade de Vida , Resultado do TratamentoRESUMO
In this study, a fluorometric and colorimetric dual-readout lateral flow immunoassay (LFIA) using antibody functionalized polydopamine-coated gold nanoparticles (Au@PDAs) as a probe was developed for the detection of carbendazim (CBD). Colloidal gold nanoparticles (AuNPs) were coated with polydopamines (PDA) by the oxidation of dopamine to synthesize Au@PDA nanoparticles. The Au@PDA nanoparticles mediated ZnCdSe/ZnS quantum dots (QDs) fluorescence quenching and recovery, resulting in a reverse fluorescence enhancement detection format of CBD. The CBD detection was obtained by the competition between the CBD and the immobilized antigen for Au@PDAs labelled antibody binding, resulting in a significant fluorescence increase and colorimetry decrease corresponded to the concentration of CBD. Dual readout modes were incorporated into the LFIA using the colorimetry signal under natural light and the fluorescence signal under UV light. The cut-off value in the mode of the colorimetric signal and fluorometric signal for CBD detection was 0.5 µg/mL and 0.0156 µg/mL, respectively. The sensitivity of LFIA of the fluorescence mode was 32 times higher than that of the colorimetry mode. There was negligible cross reactivity obtained by using LFIA for the determination of thiabendazole, benomyl, thiophanate-methyl, and thiophanate-ethyl. Consistent and satisfactory results have been achieved by comparing the results of Au@PDAs-QDs-LFIA and liquid chromatography-tandem mass spectrometry (LC-MS/MS) testing spiked cucumber and strawberry samples, indicating good reliability of the Au@PDAs-QDs-LFIA.
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Nanopartículas Metálicas , Pontos Quânticos , Benzimidazóis/farmacologia , Carbamatos/farmacologia , Cromatografia Líquida , Ouro/química , Imunoensaio/métodos , Indóis/química , Nanopartículas Metálicas/química , Polímeros/química , Pontos Quânticos/química , Reprodutibilidade dos Testes , Espectrometria de Massas em TandemRESUMO
This study investigated the presence of 18 dioxin-like and non-dioxin-like polychlorinated biphenyls (dl- and ndl-PCBs), heavy metals (Cd, Hg, Pb, and As) in Chinese mitten crabs (Eriocheir sinensis) and their distribution in Jiangsu, China. Risk assessment and source apportionment were employed for evaluating the eco-toxicological impact and human exposure. It was found that the compositions of PCBs varied spatially, suggesting different sources of pollutants, whilst PCB 28, 105, 114, and 126 were consistently found in all sample types, suggesting a common pollution source remained, and the bio-accumulation process was in effect. The total PCBs in sediment were found much higher than in water, and brown meat had the highest and most diverse PCB congeners among all tissues. The presence of heavy metals was found in all samples in descending order of As>Cd>Pb>Hg and in the order of shell>brown meat>white meat>gill for crabs. The results of risk assessment indicated that the potential carcinogenic and non-carcinogenic risks were within the acceptable range for long-term consumption of the crabs overall. However, the highest toxic equivalent (TEQ), carcinogenic, and non-carcinogenic risks were all recorded in Location C, where dl-PCB 126, 169, and As contributed to the majority of the risks. The ecological risk posed by all HMs was low, but cases of serious point source pollution have been found in the investigated regions, and risks caused by Cd individually should raise concerns. Source apportionment study revealed that the contaminants mostly originated from anthropogenic activities. Natural deposition and transportation played an important role as well.
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Braquiúros/química , Exposição Dietética , Metais Pesados , Bifenilos Policlorados , Agricultura , Animais , China , Ecossistema , Monitoramento Ambiental , Humanos , Metais Pesados/análise , Bifenilos Policlorados/análise , Medição de RiscoRESUMO
New nanocomposites, Fe3O4@Au-FITC, were prepared and explored to develop a fluorescent detection of Pb2+. The Fe3O4@AuNPs-FITC nanocomposites could be etched by Pb2+ in the presence of Na2S2O3, leading to fluorescence recovery of FITC quenched by Fe3O4@Au nanocomposites. With the increase of Pb2+ concentration, the fluorescence recovery of Fe3O4@AuNPs-FITC increased gradually. Under optimized conditions, a detection limit of 5.2 nmol/L of Pb2+ with a linear range of 0.02-2.0 µmol/L were obtained. The assay demonstrated negligible response to common metal ions. Recoveries of 98.2-106.4% were obtained when this fluorescent method was applied in detecting Pb2+ spiked in a lake-water sample. The above results demonstrated the high potential of ion-induced nanomaterial etching in developing robust fluorescent assays.
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In this report, a fluorescent sensing method for paraquat based on gold nanoclusters (AuNCs) is proposed. It was found that paraquat could quench both glutathione-capped AuNCs (GSH-AuNCs) and ß-cyclodextrin-modified GSH-AuNCs (GSH/ß-CDs-AuNCs). The modification of ß-CDs on the surface of GSH-AuNCs obviously enhanced the fluorescence intensity of GSH-AuNCs and improved the sensitivity of paraquat sensing more than 4-fold. This sensibilization was ascribed to the obvious fluorescence intensity enhancement of GSH-AuNCs by ß-CDs and the "host-guest" interaction between paraquat and ß-CDs. The fluorescence quenching was mainly due to the photoinduced energy transfer (PET) between GSH/ß-CDs-AuNCs and paraquat. With the optimized ß-CDs modification of the GSH-AuNC surfaces and under buffer conditions, the fluorescent detection for paraquat demonstrated a linear response in the range of 5.0-350 ng/mL with a detection limit of 1.2 ng/mL. The fluorescent method also showed high selectivity toward common pesticides. The interference from metal ions could be easily masked by ethylene diamine tetraacetic acid (EDTA). This method was applied to the measurement of paraquat-spiked water samples and good recoveries (93.6-103.8%) were obtained. The above results indicate that host molecule modification of fluorescent metal NC surfaces has high potential in the development of robust fluorescent sensors.
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The anti-idiotypic antibody is widely used in the field of immunology to simulate structural features or even induce the biological activity of antigens. In this study, we obtained seven anti-idiotypic single-chain variable fragments (scFv) antibodies of Cry2Aa toxin from a phage-displayed mutant library constructed using error-prone PCR technique. A mutant designated 2-12B showed the best binding ability amongst all anti-idiotypic scFv isolates to Plutella xylostella brush border membrane vesicles (BBMVs). 2-12B and Cry2Aa toxin shared a potential receptor of polycalin in P. xylostella BBMVs. Homology modeling and molecular docking demonstrated that 2-12B and Cry2Aa toxin have seven common binding amino acid residues in polycalin. Insect bioassay results suggested that 2-12 had insecticidal efficacy against P. xylostella larvae. These results indicated that the Cry2Aa anti-idiotypic scFv antibody 2-12B partially mimicked the structure and function of Cry2Aa toxin. The anti-idiotypic scFv antibody provides the basic material for the future study of surrogate molecules or new insecticidal materials.